Citation
Responses made by the salt marsh teleost Cyprinodon variegatus (Atherinomorpha: Cyprinodontidae) to live in a variable salinity environment

Material Information

Title:
Responses made by the salt marsh teleost Cyprinodon variegatus (Atherinomorpha: Cyprinodontidae) to live in a variable salinity environment
Creator:
Haney, Dennis Charles, 1962-
Publication Date:
Language:
English
Physical Description:
xi, 123 leaves : ill. ; 29 cm.

Subjects

Subjects / Keywords:
Blood ( jstor )
Erythrocytes ( jstor )
Fish ( jstor )
Hematocrit ( jstor )
Metabolism ( jstor )
Oxygen ( jstor )
Oxygen partial pressure ( jstor )
Plasmas ( jstor )
Salinity ( jstor )
Salt marshes ( jstor )
Dissertations, Academic -- Zoology -- UF
Sheepshead minnow ( lcsh )
Zoology thesis, Ph. D
Gulf of Mexico ( local )
Genre:
bibliography ( marcgt )
non-fiction ( marcgt )

Notes

Thesis:
Thesis (Ph. D.)--University of Florida, 1995.
Bibliography:
Includes bibliographical references (leaves 108-122).
General Note:
Typescript.
General Note:
Vita.
Statement of Responsibility:
by Dennis Charles Haney.

Record Information

Source Institution:
University of Florida
Holding Location:
University of Florida
Rights Management:
Copyright [name of dissertation author]. Permission granted to the University of Florida to digitize, archive and distribute this item for non-profit research and educational purposes. Any reuse of this item in excess of fair use or other copyright exemptions requires permission of the copyright holder.
Resource Identifier:
022003406 ( ALEPH )
34424550 ( OCLC )
AKP6468 ( NOTIS )

Downloads

This item has the following downloads:

ELIYC40N1_1XLF0E.xml

responsesmadebys00hane.pdf

responsesmadebys00hane_0032.txt

responsesmadebys00hane_0026.txt

responsesmadebys00hane_0131.txt

responsesmadebys00hane_0086.txt

responsesmadebys00hane_0077.txt

responsesmadebys00hane_0038.txt

responsesmadebys00hane_0128.txt

responsesmadebys00hane_0082.txt

responsesmadebys00hane_0125.txt

responsesmadebys00hane_0127.txt

responsesmadebys00hane_0117.txt

responsesmadebys00hane_0091.txt

responsesmadebys00hane_0025.txt

responsesmadebys00hane_0006.txt

AA00009024_00001.pdf

responsesmadebys00hane_0015.txt

responsesmadebys00hane_0085.txt

responsesmadebys00hane_0090.txt

responsesmadebys00hane_0028.txt

responsesmadebys00hane_0060.txt

responsesmadebys00hane_0135.txt

responsesmadebys00hane_0005.txt

responsesmadebys00hane_0037.txt

responsesmadebys00hane_0013.txt

responsesmadebys00hane_0017.txt

responsesmadebys00hane_0007.txt

responsesmadebys00hane_0078.txt

responsesmadebys00hane_0116.txt

responsesmadebys00hane_0110.txt

responsesmadebys00hane_0027.txt

responsesmadebys00hane_0081.txt

responsesmadebys00hane_0059.txt

responsesmadebys00hane_0099.txt

responsesmadebys00hane_0069.txt

responsesmadebys00hane_0122.txt

responsesmadebys00hane_0018.txt

responsesmadebys00hane_0044.txt

responsesmadebys00hane_0002.txt

responsesmadebys00hane_0111.txt

responsesmadebys00hane_0132.txt

responsesmadebys00hane_0121.txt

responsesmadebys00hane_0066.txt

responsesmadebys00hane_0074.txt

responsesmadebys00hane_0054.txt

responsesmadebys00hane_0094.txt

responsesmadebys00hane_0039.txt

responsesmadebys00hane_0118.txt

responsesmadebys00hane_0103.txt

responsesmadebys00hane_0126.txt

responsesmadebys00hane_0112.txt

responsesmadebys00hane_0004.txt

responsesmadebys00hane_0003.txt

responsesmadebys00hane_0096.txt

responsesmadebys00hane_0075.txt

responsesmadebys00hane_0016.txt

responsesmadebys00hane_0092.txt

responsesmadebys00hane_0095.txt

responsesmadebys00hane_0033.txt

responsesmadebys00hane_0134.txt

responsesmadebys00hane_0056.txt

responsesmadebys00hane_0068.txt

responsesmadebys00hane_0076.txt

responsesmadebys00hane_0012.txt

responsesmadebys00hane_0062.txt

responsesmadebys00hane_0042.txt

responsesmadebys00hane_0014.txt

responsesmadebys00hane_0043.txt

responsesmadebys00hane_0087.txt

responsesmadebys00hane_0102.txt

responsesmadebys00hane_0024.txt

responsesmadebys00hane_0097.txt

responsesmadebys00hane_0119.txt

responsesmadebys00hane_0029.txt

responsesmadebys00hane_0008.txt

responsesmadebys00hane_0061.txt

responsesmadebys00hane_0057.txt

responsesmadebys00hane_0084.txt

responsesmadebys00hane_0001.txt

responsesmadebys00hane_0045.txt

responsesmadebys00hane_0050.txt

responsesmadebys00hane_0021.txt

responsesmadebys00hane_0020.txt

responsesmadebys00hane_0046.txt

responsesmadebys00hane_0023.txt

responsesmadebys00hane_0114.txt

responsesmadebys00hane_0009.txt

responsesmadebys00hane_0065.txt

AA00009024_00001_pdf.txt

responsesmadebys00hane_0030.txt

responsesmadebys00hane_0107.txt

responsesmadebys00hane_0052.txt

responsesmadebys00hane_0093.txt

responsesmadebys00hane_0063.txt

responsesmadebys00hane_0000.txt

responsesmadebys00hane_0055.txt

responsesmadebys00hane_0113.txt

responsesmadebys00hane_0129.txt

responsesmadebys00hane_0115.txt

responsesmadebys00hane_0036.txt

responsesmadebys00hane_0035.txt

responsesmadebys00hane_0071.txt

responsesmadebys00hane_0088.txt

responsesmadebys00hane_0058.txt

responsesmadebys00hane_0080.txt

responsesmadebys00hane_0124.txt

responsesmadebys00hane_0019.txt

responsesmadebys00hane_0137.txt

responsesmadebys00hane_0049.txt

responsesmadebys00hane_0067.txt

responsesmadebys00hane_0047.txt

responsesmadebys00hane_0079.txt

responsesmadebys00hane_0133.txt

responsesmadebys00hane_0073.txt

responsesmadebys00hane_0072.txt

responsesmadebys00hane_0130.txt

responsesmadebys00hane_0101.txt

responsesmadebys00hane_0120.txt

responsesmadebys00hane_0098.txt

responsesmadebys00hane_0070.txt

responsesmadebys00hane_0034.txt

responsesmadebys00hane_pdf.txt

responsesmadebys00hane_0041.txt

responsesmadebys00hane_0031.txt

responsesmadebys00hane_0053.txt

responsesmadebys00hane_0108.txt

responsesmadebys00hane_0109.txt

responsesmadebys00hane_0064.txt

responsesmadebys00hane_0136.txt

responsesmadebys00hane_0100.txt

responsesmadebys00hane_0022.txt

responsesmadebys00hane_0089.txt

responsesmadebys00hane_0123.txt

responsesmadebys00hane_0010.txt

responsesmadebys00hane_0011.txt

responsesmadebys00hane_0048.txt

ELIYC40N1_1XLF0E_xml.txt

responsesmadebys00hane_0106.txt

responsesmadebys00hane_0051.txt

responsesmadebys00hane_0104.txt

responsesmadebys00hane_0105.txt

responsesmadebys00hane_0040.txt

responsesmadebys00hane_0083.txt


Full Text









RESPONSES MADE BY THE SALT MARSH TELEOST CYPRINODON
VARIEGATUS (ATHERINOMORPHA: CYPRINODONTIDAE) TO LIFE IN A
VARIABLE SALINITY ENVIRONMENT













By


DENNIS CHARLES HANEY


A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL
OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT
OF THE REQUIREMENTS FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY


UNIVERSITY OF FLORIDA


1995


/-l /" /




A £ PCrfC?
RESPONSES MADE BY THE SALT MARSH TELEOST CYPR1NODON
VARIEGATUS (ATHERINOMORPHA: CYPRINODONTIDAE) TO LIFE IN A
VARIABLE SALINITY ENVIRONMENT
By
DENNIS CHARLES HANEY
A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL
OFTHE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT
OF THE REQUIREMENTS FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY
UNIVERSITY OF FLORIDA


ACKNOWLEDGMENTS
The work described in this dissertation was the focus of my activities for many
years. Throughout this time I have been fortunate to meet and interact with an extraordinary
variety of people, many of whom played integral roles in the successful completion of this
dissertation.
I would first like to acknowledge the assistance of the members of my graduate
committee: Frank Nordlie, Steve Walsh, Brian McNab, Harvey Lillywhite, Ken Sulak, and
Tom Crisman. I would also like to thank a founding member of my committee, Michelle
Wheatly, who, unfortunately for me, left the University of Florida before I was able to
complete my dissertation. All of these people were instrumental in the completion of this
work. In particular, I owe an incredible debt of gratitude to the chair of my committee, Frank
Nordlie. His unfailing friendship, help, and guidance over the years was truly an inspiration
to me, and I can safely say I would not have made it through to the end without him. Steve
Walsh also deserves special recognition for his much-needed assistance. Steve has been a
close friend and colleague for many years, and I am proud to be the first student to complete
a dissertation with Steve as a member of his committee!
So many members of the Zoology Department have helped out that its difficult to
know where to begin. Brent and Sylvia Palmer, John Matter, Frank Hensley (my fusiform
twin!), and Lou Somma were all invaluable in helping me to maintain good mental health
over the years. We spent many hours trying to figure out life, the universe, and everything.
We also had lots of fun looking for unsuspecting fish and herps. Lisa Gregory has been a
friend I could always count on no matter what! John Binello was my trusted field
companion. We spent many days tramping through the salt marsh (and other bodies of
11


water) in search of numerous fishes including the wily pupfish, sailfin molly, Florida
flagfish, and fat sleeper. Thanks also go to Leo Nico, Adele Hensley, Pam Fuller, Paula
Cushing, Patricia Harrison, Lianna Jarecki, Becky Thompson, Vince DeMarco, Kevin
Baldwin, Ellen Burroughs, Chris Kardish, Mark Hostetler, Doug Weaver, Frank Jordan,
John Anderson, Carol Binello, and everyone else from Zoology!
For the past 4+ years I have worked with many people at the Department of the
Interior's Gainesville laboratory (presently the National Biological Service, formerly U.S.
Fish and Wildlife Service). While working full-time for the past few years certainly slowed
down my progress towards finishing my Ph.D., the extra time was worth it (mostly!). I
formed many new friendships and learned lots of things I wouldn't have otherwise. Jim
Williams showed me that freshwater clams were actually kind of neat. Noel Burkhead
helped remind me that fish were still much cooler! Les Parker, Jayne Brim-Box, and I had
great fun diving in zero visibility water. Howard Jelks, Gary Hill, Ann Foster, Tina Yanchis,
Rob Whiteford, and Bill Stranghoener were all inspiring at one time or another. Special
thanks go to my office mates, Leslie Straub and Cindy Timmerman, for putting up with me
for so long. Cindy especially has been a great friend and confidante.
Last, but certainly not least, I want to thank my family for supporting me all these
years. Mom and Dad never doubted me, though it seemed like I would never finish. Their
encouragement really did help! My brother Scott helped to show me you really can finish a
dissertation and be successful afterwards. For this, and everything else, I can never thank
him enough.
iii


TABLE OF CONTENTS
LIST OF TABLES vi
LIST OF FIGURES v
ABSTRACT x
CHAPTERS
1 INTRODUCTION 1
Experimental Animal 7
Questions 10
Study Site 11
2 INFLUENCE OF ENVIRONMENTAL SALINITY ON ROUTINE
METABOLIC RATE AND CRITICAL OXYGEN TENSION OF
CYPRINODON VARIEGATUS 12
Introduction 12
Methods 14
Results 18
Discussion 31
3 INFLUENCE OF SIMULATED TIDAL CHANGES IN AMBIENT
SALINITY ON ROUTINE METABOLIC RATE IN CYPRINODON
VARIEGATUS 39
Introduction 39
Methods 40
Results 46
Discussion 46
4 INFLUENCE OF A FLUCTUATING SALINITY REGIME ON
OSMOREGULATION IN CYPRINODON VARIEGATUS 55
Introduction 55
Methods 56
Results 58
Discussion 66
5 INFLUENCE OF ENVIRONMENTAL SALINITY ON BLOOD
OXYGEN LEVELS OF CYPRINODON VARIEGATUS 70
Introduction 70
Methods 71
IV


Results 73
Discussion 79
6 SUMMARY AND CONCLUSIONS 87
APPENDICES 91
1 CRITICAL OXYGEN TENSION FIGURES 91
2 FIELD MEASUREMENTS 102
LITERATURE CITED 108
BIOGRAPHICAL SKETCH 123
v


LIST OF TABLES
Table page
1-1. Phylogenetic classification of the cyprinodontiform fishes (modified from
Parenti, 1981) 8
2-1. Relationships of routine metabolism (RMR), critical oxygen tension (Pc), and
slope in the conformation region at a series of ambient salinities. Values are given
as means se 19
2-2. Measurements of oxygen concentration (mg Ll), salinity (ppt), and temperature
(C) taken at four sites in the Cedar Key area from June 1990 through June 1991
(see text for details on location of sites). Values are given as means, se (in
parentheses), sample size 32
3-1. Acclimation and final salinities used in simulated tidal change study 42
3-2. Mean routine metabolism (mg Cb h'l) before (acclimation salinity) and
following (final salinity) a simulated tidal change. Values are given as means se.
Groups exhibiting a significant change in metabolism are indicated with an
asterisk 47
4-1. Salinity trials used in cyclical salinity study. The group maintained at 30 ppt was
split into three groups following cycle 10 (day 20); groups Cq, C, and Q (see text
for details) 58
4-2. Results of salinity fluctuations experiment. Values in the top row of each cell
represent hematocrit measurements (% erythrocytes), values in bottom row of each
cell represent plasma osmolality measurements (mOsm kg'l). Sample sizes are
n=5 for each cell. All values are expressed as means se. See text for explanation
of group abbreviations 60
5-1. Hematocrit (Hct), hemoglobin concentration ([Hb]), erythrocyte count (RBC),
mean corpuscular hemoglobin (MCH), mean corpuscular volume (MCV), and
mean corpuscular hemoglobin concentration (MCHC) as a function of salinity for
Cyprinodon variegatus. All values are expressed as means se. See text for
further details on blood indices 74
vi


LIST OF FIGURES
Figure page
2-1. Mean adjusted routine metabolic rates (RMR) over a range of salinities in
Cyprinodon variegatus (metabolic rates were mass-adjusted using an analysis of
covariance; bars indicate se; numerical values above the points in the figure
indicate sample sizes at each salinity) 21
2-2. Relationship between mean adjusted routine metabolic rates (RMR) and mean
plasma osmolality over a range of salinities in Cyprinodon variegatus (metabolic
rates were mass-adjusted using an analysis of covariance; bars indicate se;
plasma osmolality data from Nordlie, 1985) 23
2-3. Mean critical oxygen tension (Pc) measurements over a range of salinities in
Cyprinodon variegatus (bars indicate se; numerical values above the points in
the figure indicate sample sizes at each salinity) 26
2-4. Relationship between mean adjusted routine metabolic rates (RMR) and critical
oxygen tensions (Pc) over a range of salinities in Cyprinodon variegatus
(metabolic rates were mass-adjusted using an analysis of covariance; bars
indicate se) 28
2-5. Relationship between mean critical oxygen tensions (Pc) and mean plasma
osmolality over a range of salinities in Cyprinodon variegatus (bars indicate se;
plasma osmolality data from Nordlie, 19&5) 30
2-6. Relationship between mean adjusted routine metabolic rates (RMR) and mean
plasma osmolality over a range of salinities in Adinia xenica (metabolic rates were
mass-adjusted using an analysis of covariance; bars indicate se; numerical
values above the points in the figure indicate sample sizes at each salinity) 35
3-1. Schematic diagram of respirometry apparatus used for routine metabolism
experiments. See text for detailed description of system 44
3-2. Results of metabolic trials where salinity was increased over the course of the
trial. Bars represent groups listed in Table 3-2 for which final salinity was greater
than initial salinity. The height of each bar signifies the magnitude of the salinity
change for each metabolic trial and the asterisk indicates at which of the salinities
(for each metabolic trial) the routine metabolic rate (RMR) was highest. The x axis
has no scale and serves only to visually separate groups 49
vii


3-3. Results of metabolic trials where salinity was decreased over the course of the
trial. Bars represent groups listed in Table 3-2 for which final salinity was less
than initial salinity. The height of each bar signifies the magnitude of the salinity
change for each metabolic trial and the asterisk indicates at which of the salinities
(for each metabolic trial) the routine metabolic rate (RMR) was highest. The x axis
has no scale and serves only to visually separate groups 51
4-1. Mean plasma osmolality values measured for groups experiencing decreases in
salinity during the course of the experiment. Group designations are as follows:
Di, salinity fluctuated between 30 ppt and 2 ppt; D2; salinity fluctuated between
30 ppt and 10 ppt; D3, salinity fluctuated between 30 ppt and 20 ppt; Cq; salinity
constant at 30 ppt for days 0-20, decreased to 2 ppt following subsample on day
20; C, salinity constant at 30 ppt. Sample sizes are n=5 for each group. See
text for details of experimental procedure 62
4-2. Mean plasma osmolality values measured for groups experiencing increases in
salinity during the course of the experiment. Group designations are as follows:
Ij, salinity fluctuated between 30 ppt and 40 ppt; W, salinity fluctuated between 30
ppt and 50 ppt; I3, salinity fluctuated between 30 ppt and 60 ppt; Cj; salinity
constant at 30 ppt for days 0-20, increased to 60 ppt following subsample on day
20; C, salinity constant at 30 ppt. Sample sizes are n=5 for each group. See text
for details of experimental procedure 64
5-1. Mean erythrocyte (RBC) count over a range of salinities in Cyprinodon
variegatus (bars indicate se; numerical values above the points in the figure
indicate sample sizes at each salinity) 76
5-2. Mean hemoglobin concentration ([Hb]) over a range of salinities in Cyprinodon
variegatus (bars indicate se; numerical values above the points in the figure
indicate sample sizes at each salinity) 78
5-3. Mean hematocrit (Hct) over a range of salinities in Cyprinodon variegatus (bars
indicate se; numerical values above the points in the figure indicate sample sizes
at each salinity) 81
A1-1. Plot indicating the calculation of the critical oxygen tension (Pc) for an
individual Cyprinodon variegatus in water at 0 ppt 93
A1-2. Plot indicating the calculation of the critical oxygen tension (Pc) for an
individual Cyprinodon variegatus in water at 50 ppt 95
A1-3. Plot indicating the calculation of the critical oxygen tension (Pc) for an
individual Cyprinodon variegatus in water at 100 ppt 97
A1-4. Generalized critical oxygen tension (Pc) plots at each salinity used in the Pc
experiments. Plots were produced by using the mean Pc, mean routine metabolic
rate (RMR), and mean slope in the conformation region for each salinity group..
viii
99


A2-1. Oxygen concentration (mg L"l), salinity (ppt), and temperature (C) at four
sites in the Cedar Key area taken between June 1990 and June 1991. Values are
given as means, bars indicate se. a) Measurements taken on the bottom at site 1;
b) measurements taken on the surface at site 1; c) Measurements taken on the
bottom at site 2; d) measurements taken on the surface at site 2; e) Measurements
taken on the bottom at site 3; 0 measurements taken on the surface at site 3; g)
Measurements taken on the surface at site 4
104
IX


Abstract of Dissertation Presented to the Graduate School
of the University of Florida in Partial Fulfillment of the
Requirements for the Degree of Doctor of Philosophy
RESPONSES MADE BY THE SALT MARSH TELEOST CYPRINODON
VARIEGATUS (ATHERINOMORPHA : CYPR1NODONT1DAE) TO LIFE IN A
VARIABLE SALINITY ENVIRONMENT
By
DENNIS CHARLES HANEY
December, 1995
Chairman: Frank Nordlie
Major Department: Zoology
Cyprinodon variegatus, a common coastal resident of the western Atlantic Ocean
and Gulf of Mexico, lives in ambient salinities ranging from freshwater to 142 ppt. Fish
used in this study were obtained from a Gulf of Mexico salt marsh near Cedar Key, Florida.
In a steady-state experiment, routine metabolic rate (RMR) and critical oxygen tension (Pc)
were determined at salinities ranging from 0 tolOO ppt. Salinities between 0 and 40 ppt had
little influence on RMR or Pc. However, at salinities above 40 ppt, RMR declined, and Pc
increased. The reduction in RMR and rise in Pc correlates with a reduced ability of C.
variegatus to osmoregulate effectively at high salinities. The variations in RMR and Pc at
high salinities suggests that C. variegatus responds by reducing energy expenditures,
effectively increasing the time that individuals can tolerate hypersaline conditions. The
metabolic patterns of C. variegatus as influenced by simulated tidal changes in salinity were
then measured, with RMR unaffected by changes in salinity between 2 and 40 ppt.
However, at extremely high or low salinities metabolism was affected by changes in ambient
x


salinity. Individuals of C. variegatus responded to fluxes at salinity extremes by reducing
general activity and energy expendituresessentially waiting for conditions to return to
normal, where they responded by increasing metabolic activity. Cyprinodon variegatus
efficiently regulated plasma osmolality, even when fishes were exposed to large fluctuations
in salinity. However, prior exposure to salinity fluctuations did impart an osmoregulatory
advantage. Fish previously exposed to large salinity fluctuations regulated plasma
osmolality better than fish that previously experienced no or small changes in salinity.
Increasing salinity had a greater impact on osmoregulation than did decreasing salinity.
Salinity also had a significant effect on blood oxygen carrying capacity in C. variegatus,
although differences were only noted at the very highest (60 80 ppt) and lowest (0 ppt)
salinities tested. Oxygen carrying capacity was highest in the group acclimated to 0 ppt.
Erythrocyte count was the most consistent indicator of the influence of salinity on blood
oxygen, with hematocrit the least consistent measure.
xi


CHAPTER 1
INTRODUCTION
Salinity is a crucial physicochemical factor that exerts an important influence on
aquatic life, particularly on estuarine and salt marsh organisms that are exposed to
unpredictable salinity fluctuations diumally and seasonally. Salt marshes are intertidal beds
of rooted vegetation that are alternately flooded and exposed by rising and falling tides.
Vegetation on higher ground develops a complex network of branching channels through
which water, nutrients, and aquatic organisms move during the tidal cycle. Intertidal salt
marshes are inhospitable yet interesting environments due to their unstable water levels and
to the extreme variability of their physical, chemical, and biological processes (Cooper,
1974; Adam, 1990; Allen and Pye, 1992).
Factors that may influence salinity in salt marshes include precipitation, wind,
frequency and extent of tides, shoreline height, and coastal topography (Remane and
Schlieper, 1971; Wheatly, 1988). Due to the harshness of the physical environment, salt
marshes tend to have low species diversity, but often high abundance of selected species
(McCIusky, 1989; Dunson and Travis, 1994).
Whereas variable salinity habitats are also characterized by fluctuations in other
environmental factors, such as temperature, dissolved oxygen, and pH (Vemberg, 1983;
Wheatly, 1988), the distribution and abundance of fishes in these habitats is largely
determined by salinity (McCIusky, 1989; Davenport and Sayer, 1993; Gill and Potter,
1993). Not surprisingly, animals respond to fluctuations in salinity in complex ways.
Salinity affects osmoregulation, ventilation, metabolism, acid-base balance, growth,
reproduction, development, and other biological processes (Wheatly, 1988). Some of the
primary responses of telcosts to changes in salinity are reviewed here.
1


2
Avoidance is the first line of defense to variable salinity conditions. If behavioral
responses do not sufficiently minimize exposure to variable salinities, aquatic organisms
must rely on physiological and biochemical responses to tolerate environmental changes
(Beitinger and McCauley, 1990). This may be by passive tolerance (osmoconforming) or
active osmoregulation (Truchot, 1987; McClusky, 1989). Only fishes that are capable of
osmoregulation can tolerate wide changes in salinity, and these euryhaline teleosts will be
the focus of the rest of this review.
The basic patterns of osmoregulation in fishes have been extensively reviewed in
recent years (Parvatheswararao, 1970; Eddy, 1982; Evans, 1984; Zadunaisky, 1984;
Kamaky, 1986; Foskett, 1987; Pisam and Rambourg, 1991; Ventrella et al., 1992; Evans,
1993; McCormick, 1994; Wood and Marshall, 1994). Euryhaline teleosts regulate their
blood osmolality at about one-third the concentration of seawater (35 ppt), and thus face
severe osmotic problems whether in freshwater (0 ppt) or seawater. Body fluids of a teleost
in freshwater are hyper-osmotic to the external environment, whereas in seawater they are
hypo-osmotic. Thus, euryhaline fish possess mechanisms for osmoregulating in both
hyper-osmotic and hypo-osmotic conditions.
Teleosts in seawater are susceptible to a loss of body water to the external
environment, and balance water loss by actively drinking large amounts of seawater.
However, both water and salts are absorbed together across the gut. Ingested excess salts
are actively excreted, divalent ions mostly in urine and feces, and monovalent ions by the
gills.
Active excretion of salts by the gills takes place via chloride cells (Zadunaisky, 1984;
Kamaky, 1986; Foskett, 1987; Pisam and Rambourg, 1991). Chloride cells in scawatcr-
acclimatcd fish are located at the base of the secondary gill lamellae, and are large,
columnar-shaped cells usually extending from the basal epithelium to the external
environment. They arc characterized by numerous mitochondria (for this reason they arc
often referred to as "mitochondria rich cells") and an extensive tubular reticulum continuous


3
with the basolateral membrane (Pisam and Rambourg, 1991). The transport enzyme
Na+,K+-ATPase is restricted to this tubular system and to the basolateral membrane. This
ion pump creates a large Na+ gradient (low in the chloride cell cytoplasm) which drives a
NaCl co-transporter by which Cl' enters the cell. The Cl" accumulates sufficiently in the cell
such that it is able to exit to the external environment across the apical membrane, with Na+
following passively down its electrochemical gradient between adjacent chloride cells
(Evans, 1993; McCormick, 1994; Wood and Marshall, 1994).
In freshwater, teleosts continuously face an efflux of salts and an influx of water.
Hence, their osmoregulatory response is to actively transport salts from the external
environment via the gills, to avoid drinking water, and to excrete copious amounts of dilute
urine. Our present understanding of the mechanisms involved in the uptake of Na+ and Cl'
from the environment in freshwater teleosts is somewhat unclear and incomplete. It appears
that Cl" is actively taken up, with Na+ uptake being a passive process. Whether this takes
place via a freshwater-type chloride cell, or through interactions with acid-base balance is
unclear, as evidence exists for both (Wood and Marshall, 1994).
What is unique about osmoregulation in euryhaline fish is not that they possess a
structurally distinct osmoregulatory mechanism, but that they have the ability to function
efficiently in variable salinities. They must not only be able to hypo- and hyper-
osmoregulate, but they must be able to alter their pattern of osmoregulation quickly if they
live where salinity rapidly fluctuates.
Osmoregulation is controlled largely by the endocrine system. In teleosts, hormones
known to exert osmoregulatory effects include prolactin, growth hormone, cortisol,
angiotensin II, arginine vasotocin, atrial natriuretic peptide, thyroid hormones, urotensins,
vasoactive intestinal peptide, insulin, calcitonin, catecholamines and sex steroids (Jenkins,
1981; Bern and Madsen, 1992; Lcloup and Lcbcl, 1993; Sakamoto et al., 1993;Takei, 1993;
McCormick, 1994). The hormone most involved with osmoregulation under freshwater
conditions is prolactin, with arginine vasotocin, urotensin II, catecholamines, and atrial


4
natriuretic peptide also playing roles (Hirano et al., 1987; Bern and Madsen, 1992; Takei,
1993). Control of osmoregulation in seawater is largely via the effects of cortisol and
growth hormone, together with thyroid hormones, angiotensin II, vasoactive intestinal
peptide, atrial natriuretic peptide, and urotensin I (Balment etal., 1987; Bern and Madsen,
1992; Takei, 1993). Although the exact mechanisms and interactive effects of many of these
hormones are unclear, it is well established that both rapid and long term control of
osmoregulation under hyper-osmotic and hypo-osmotic conditions is mediated via the
endocrine system.
Osmoregulation is not the only physiological process affected by salinity. Salinity
adaptation is a complex event that involves a number of physiological and behavioral
responses, including energetics. Many studies have shown that salinity influences
metabolism of fishes, but little agreement exists with respect to the magnitude or direction of
these effects (Nordlie, 1978; Febry and Lutz, 1987; Nordlie et al., 1991; Swanson, 1991;
Morgan and Iwama, 1991; Kirschner, 1993). One reason for a lack of agreement among
studies is that many have focused on the energetic costs associated with osmoregulation,
while ignoring other related physiological and behavioral processes. For example, many
studies have used measured differences in metabolism at different salinities to represent
differences in osmoregulatory costs. The rationale is that osmoregulation is an energetically
costly event, with the energetic costs being related to the osmotic gradient between the fish
and its environment (e.g., Madan Mohan Rao, 1968; Muir and Niimi, 1972; Furspan et al.,
1984; Zadunaisky, 1984). However, this is probably an oversimplification, since other
factors, such as activity, food intake, and permeability changes may also be influenced by
salinity, and in turn, affect metabolism (Swanson, 1991). For this reason metabolic
measurements represent the overall costs associated with living in a particular salinity
environment, with comparisons among salinities reflecting these total costs, not simply the
cost of osmoregulation.


5
Several attempts have been made to categorize the patterns of metabolic responses
of teleosts to altered salinities (Kinne, 1967; Remane and Schlieper, 1971; Nordlie, 1978;
Morgan and Iwama, 1991). One commonly observed pattern is that the lower rates of
metabolism in response to salinity are associated with environments for which the species
(and life stage) are presumably best adapted for, and in which they are normally found.
Teleosts that inhabit widely variable salinity environments are uniquely characterized by
having metabolic rates that are constant over a wide range of salinities. This allows
euryhaline teleosts to avoid the large metabolic costs normally associated with physiological
adjustment to changing salinity.
Another physicochemical parameter that may act synergistically with salinity is
dissolved oxygen. The dissolved oxygen content of many bodies of water is subject to large
natural fluctuations. This is especially true in shallow water, such as salt marshes, where
chronic and/or periodic hypoxia may be common (Renaud, 1985; Dejours, 1987;
Toulmond, 1987; Graham, 1990). Variations in oxygen can dictate the distribution of some
species in aquatic ecosystems (Boutilier, 1990). Changing environmental salinity may
directly influence respiratory function by affecting both the oxygen solubility in water
pumped over the gills and the solubility of gases dissolved in plasma. Changes in the ionic
composition of the body fluids could also interact with oxygen to influence tolerance to
variable salinity conditions (Truchot, 1987).
Exposure of fish to reduced oxygen tensions initiates physiological responses
mostly directed at increasing the amount of oxygen available to the tissues (Boutilier et al.,
1988). The transfer of oxygen from the environment to the tissues can be characterized as a
series of processes. Gill ventilation is the first step, followed in turn by branchial diffusion,
blood oxygen transport, and diffusion into the tissues (the last of which, due to a lack of
available information, will not be discussed further) (Perry and McDonald, 1993).
Gill ventilation is a function of the frequency and depth of breathing, and is
normally increased when low oxygen tensions are encountered. While ventilation docs not


6
appear to be directly limiting to the uptake of oxygen, substantial cost is involved with
increased ventilatory pumping, which ultimately means that any additional oxygen acquired
is used to fuel the ventilatory apparatus itself (Boutilier et al., 1988; McMahon, 1988;
Cameron, 1989; Perry and McDonald, 1993).
Two processes are largely available to increase branchial oxygen diffusion:
increases in functional gill surface area and increases in the mean water to blood oxygen
partial pressure gradient. This tradeoff is particularly important in regard to salinity, as fish
in waters of low oxygen tension must balance the advantage of maximizing branchial
oxygen diffusion with a disadvantage in osmoregulation due to the accompanying increases
in ion and water exchange (Perry and McDonald, 1993). Increasing blood gas transport is
likely the primary route used by most fish to increase the amount of oxygen delivered to the
tissues. Oxygen transport by the blood in teleosts depends on the respiratory pigment
hemoglobin. Blood oxygen transport is normally increased by increasing the concentration
of hemoglobin, increasing the number of erythrocytes in circulation, and/or by adjusting the
affinity of hemoglobin for oxygen (Davis, 1975; Wells et al., 1989; Jensen et al., 1993;
Perry and McDonald, 1993).
All of the processes described above can be modified to optimize oxygen transport
under a variety of environmental conditions. One additional strategy that can be utilized in
conjunction with the above is the lowering of metabolism in concert with reductions in
oxygen. This potentially minimizes the impact of the lowered oxygen tension, but also
reduces aerobic metabolism and therefore the amount of energy available for physiological
processes.
Most fish would be described as metabolic oxygen regulators, as they maintain a
constant metabolic rate over a range of oxygen tensions extending downward from
atmospheric levels to some low level that has been defined as the critical oxygen tension
(Pc). Below the Pc, metabolism is dependent upon oxygen tension, and decreases linearly
with decreases in oxygen. The Pc was probably first documented by Hall (1929), but was


7
not considered an important index for fishes until formalized by Fry (1947). However, this
is an extremely important variable relating to habitat selection and overall metabolic patterns
of fishes, and calculations of the Pc have since been made for a number of species under a
variety of environmental conditions.
Experimental Animal
The subject of this study was the sheepshead minnow, Cyprinodon variegatus.
Cyprinodon variegatus is a member of the family Cyprinodontidae, a large and diverse
family containing over 650 species in 80 genera (Parenti, 1981; Parker and Komfield, 1995;
see Table 1-1). Members of this family are found in fresh, brackish, and salt water, and
distributed pantropically as well as throughout North America. Cyprinodon variegatus is
the type species for both the genus Cyprinodon and the family Cyprinodontidae.
The genus Cyprinodon comprises a group of approximately 30 species of small,
oviparous fishes commonly known as pupfishes. They exhibit remarkable tolerance to
harsh environmental conditions (Miller, 1981) occurring throughout North and Central
America, the Caribbean Sea, and Venezuela (Darling, 1976; Turner and Liu, 1977; Parenti,
1981; Duggins et al., 1983; Barns and Wohlgemuth, 1993). The genus is characterized by
limited genetic divergence and few unique alleles, even in morphologically distinct species
(Kodric-Brown, 1989). Much of the research on the genus has focused on species
inhabiting North American deserts, where the largest concentration of species occurs. Most
of these desert species have small, allopatric distributions (Miller, 1981; Duggins et al.,
1983). Members of this genus also occur in the coastal brackish and marine waters of
eastern North America, where C. variegatus is the dominant pupfish species.
Cyprinodon variegatus is the only pupfish species having an extensive geographic
range. It is found along the Atlantic coast from Massachusetts to the Florida Keys, and
throughout the Gulf of Mexico. A disjunct population occurs along the Yucatan peninsula
(Johnson, 1974; Darling, 1976; Duggins et al., 1983). Populations arc also located in the


8
Table 1-1. Phylogenetic classification of the cyprinodontiform fishes (modified from
Parenti, 1981).
Order Cyprinodontiformes
Suborder Aplocheiloidei
Suborder Cyprinodontoidei
Section 1
Family Profundulidae
Section 2
Division 1
Family Fundulidae
Division 2
Superfamily Poecilioidea
Family Anablepidae
Family Poeciliidae
Superfamily Cyprinodontoidca
Family Goodeidae
Family Cyprinodontidae
Subfamily Cyprinodontinae
Tribe Orestiini
Genus Orestias
Genus Kosswigichthys
Genus Aphanius
Tribe Cyprinodontini
Genus Cyprinodon
Genus Megupsilon
Genus Jordanella
Genus Floridichthys
Genus Cualac


9
Bahamas, West Indies, and Cuba (Duggins et al., 1983). Cyprinodon variegatus has also
been introduced into several areas, where they have negatively impacted native pupfish
species (Echelle and Echelle, 1987; Echelle and Connor, 1989; Kodric-Brown, 1989; Wilde
and Echelle, 1992). Genetic variability within C. variegatus is as large as the amount of
genetic divergence within the entire genus Cyprinodon, with most of this genetic divergence
occurring in populations north of Cape Hatterasvery little genetic divergence is displayed
among southern populations (Darling, 1976; Schwartz et al., 1990).
Cyprinodon variegatus is a numerically dominant and ecologically important
species throughout most of its range, especially in salt marsh and estuarine waters (Kilby,
1955; Simpson and Gunter, 1956; Relyea, 1975; Naughton and Saloman, 1978;
Subrahmanyam and Coultas, 1980; Stout, 1985; Nelson, 1992; Ross and Doherty, 1994).
Due to its importance, C. variegatus has been an important research animal in diverse
disciplines. These include investigations on the species behavior (Itzkowitz, 1974; Itzkowitz,
1978; Mettee and Beckham, 1978; Itzkowitz, 1981, Dwyer and Bculig, 1991), ecology (Doll
and Bast, 1969; Martin, 1970; Martin, 1972; Able, 1976; Harrington and Harrington, 1982;
Fyfe, 1985; Shipley, 1991; Avila etal., 1992; Wright et al., 1993), evolution (Elder and
Turner, 1994), life history (Warlcn, 1964; Dc Vlaming et al., 1978; Berry, 1987; Able, 1990;
Echelle and Echelle, 1994), physiology (Martin, 1968; Karnaky et al., 1976;
Subrahmanyam, 1980; Nordlie, 1985; Barton and Barton, 1987; Nordlie, 1987; Peterson
and Gilmore, 1988; Nordlie and Walsh, 1989; Peterson, 1990; Pnce et al., 1990; Nordlie et
al., 1991; Dunson et al., 1993), and reproduction (Raney etal., 1953; Warlen, 1964; Berry,
1987; Kodric-Brown, 1987; Conover and DcMond, 1991). The species has also been used
in voluminous toxicology experiments because of its extreme hardiness (e.g., Schimniel and
Hansen, 1975; Hawkins et al., 1984; Battalora et al., 1985; Linton, 1992).
Cyprinodon variegatus has been called "the toughest fish in North America"
(Gunter, 1967) due to its extreme tolerance of harsh environmental conditions. It is found in
waters ranging from freshwater (Ager, 1971; Johnson, 1974; Kushlan, 1980) to salinities of


10
142 ppt (Simpson and Gunter, 1956), although it typically inhabits brackish water and
coastal salt marshes. Cyprinodon variegatus is tolerant of temperatures ranging from about
1 C (Berry, 1987), to 41 C (Strawn and Dunn, 1967), and oxygen levels approaching
anoxia (Odum and Caldwell, 1955). Thus, it is an exceedingly useful experimental subject
for studying how teleost species respond to harsh environmental conditions.
Although certain organisms can withstand greater changes in environmental
conditions than others, the ability to respond to natural environmental changes is a basic
characteristic of all living systems. Unfortunately, the terminology used to describe these
responses is not uniform. Various researchers have attempted to define the terms adaptation,
acclimation, acclimatization, and accommodation (e.g., Prosser, 1955; Kinne, 1962; Prosser,
1975; Smit, 1980; Fontaine, 1993). I will use the term "adaptation" in its broadest sense,
defining it as a modification of the characteristics of an organism that facilitate an enhanced
ability to survive and reproduce in a particular environment. In this way I recognize that
adaptations involve both genetic and physiological (phenotypic) components, while not
attempting to separate these components from one another. The term acclimation will be
used as defined by Prosser (1975), where compensatory changes are measured following
changes in single environmental variables.
Questions
This study was designed to examine some of the costs to C. variegatus associated
with living in variable salinity environments. Specifically, I asked the following questions:
(1) What are the metabolic costs associated with different ambient salinities? (2) How does
salinity influence the energetic response at low oxygen tensions? (3) What is the
osmoregulatory response to variable salinity environments? (4) How docs salinity influence
blood oxygen levels? These questions were tested by the following: measurement of
metabolism in C. variegatus fully acclimated to a wide range of experimental salinities;
measurement of the critical oxygen tension in C. variegatus fully acclimated to the same


11
range of salinities; measurement of metabolism prior to, and following, simulated tidal
changes in salinity; monitoring of plasma osmolality in C. variegatus exposed to a group of
different cycling salinity regimes; and measurement of hemoglobin concentration,
erythrocyte count, and hematocrit in C. variegatus acclimated to a wide range of ambient
salinities.
Study Site
Fish used in this study were collected from tidal marshes of the Gulf of Mexico
near Cedar Key, Florida. The shore in the Cedar Key area is classified as a zero energy
sector in which wave energy is dampened over the broad, shallow limestone plateau of the
Gulf of Mexico bottom (Stout, 1985). This results in a wide intertidal zone along the coast.
Furthermore, the coastal physiography is extremely diverse due in large part to irregularities
in the shore line of the mainland, to the presence of numerous islands and oyster bars in the
tidal area, and to the maze of intertidal and subtidal creeks and channels (Kilby, 1955). No
significant sediment sources are found in this area, and tides occur on a semi-diurnal basis.
The dominant emergent vegetation in the area is Spartina alterniflora, with the salt marshes
dominated by Juncus roemerianus. Fish communities of the Juncus marsh are dominated
by atheriniforms, with C. variegatus, Fundulus similis, and Poecilia latipinna making up
50-90% of the catch throughout most of the year (Kilby, 1955; Simpson and Gunter, 1956;
Stout, 1985; pers. obs.).


CHAPTER 2
INFLUENCE OF ENVIRONMENTAL SALINITY ON ROUTINE METABOLIC RATE
AND CRITICAL OXYGEN TENSION OF CYPRINODON VARIEGATUS
Introduction
Most fishes are capable of tolerating only a narrow range of salinities. However,
some fishes live in areas that experience frequent variations in salinity. These euryhaline
species possess important physiological and behavioral mechanisms that enable them to
survive in variable salinity environments. One such fish is the sheepshead minnow,
Cyprinodon variegatus. This species ranges along most of the Atlantic coast of the U.S.,
throughout the Gulf of Mexico, and disjunctly along the Yucatan peninsula (Johnson, 1974;
Darling, 1976; Duggins et al., 1983). It typically inhabits brackish water coastal salt
marshes that undergo frequent salinity fluctuations. Cyprinodon variegatus is capable of
tolerating salinities ranging from 0 ppt (Ager, 1971; Johnson, 1974; Kushlan, 1980) to 142
ppt (Simpson and Gunter, 1956).
This study was designed to examine the metabolic response of C. variegatus over a
variety of environmental salinities. Whereas energetic responses to a number of variables
including temperature, body mass, oxygen, and activity level have been well studied, the
influence of salinity on metabolism of fishes has received less attention. Most previous
studies have found that salinity docs affect the energetics of fishes. Unfortunately, the
magnitude and/or direction of these effects are equivocal (Febry and Lutz, 1987; Morgan
and Iwama, 1991; Nordlic et al., 1991; Swanson, 1991). This has led several authors to
categorize the general patterns of metabolic responses to altered salinities (c.g., Nordlic,
1978; Morgan and Iwama, 1991).
12


13
One general pattern of fishes that inhabit variable salinity environments is a stable
metabolic rate over a range of salinities, with the range of salinities most commonly tested
between freshwater (0 ppt) and seawater (35 ppt) (Morgan and hvama, 1991). Physiological
stability enables such fish to have a euryhaline existence that is unfettered by large
metabolic costs associated with adjustment to salinity change. However, there are relatively
few studies of the metabolic response of euryhaline fishes over an even wider range of
salinities that they encounter in their natural habitats.
Fish metabolism is also strongly influenced by the partial pressure of oxygen
(PO2). Metabolism of fishes is independent of PO2, as they maintain a constant metabolic
rate over a range of PO2 extending downward from high atmospheric levels to some lower
level defined as the critical oxygen tension (Pc). Below the Pc (conformation region),
metabolism depends on oxygen tension and decreases linearly with decreases in oxygen
(Fry, 1947). Oxygen consumption declines at the Pc because the gas exchange system can
no longer supply both the extra demands of the respiratory system and the oxygen
demands of the tissues (Hughes, 1964). Since Pc is a useful metabolic parameter,
calculations of the Pc have been made for a number of fishes under a variety of
environmental conditions (e.g., Hall, 1929; Tang, 1933; Ultsch et al., 1978; Ott et al., 1980;
Subrahmanyam, 1980; Donnelly and Torres, 1988; Rantin et al., 1992; Nonnotte et al.,
1993). However, the influence of ambient salinity on the Pc has not been specifically
studied. This is surprising since salinity and oxygen are important abiotic factors that may
act synergistically in affecting metabolism.
This study examined the influence of a wide range of environmental salinities on
routine metabolic rate (RMR) and Pc in C. variegatus. I hypothesized that both RMR and
Pc would be unaffected by alterations in salinity over the range of salinities commonly
encountered in natural habitats of C. variegatus. Salinities outside this range, but within the
range known to be tolerated, were predicted to cause increases in both parameters.


14
Methods
Fish used in this study were obtained by seining canals and ditches in the salt marsh
near Cedar Key, Florida (Gulf of Mexico). Specimens were transported back to the
laboratory in 19 L carboys containing water from the collection site. Upon arrival at the
laboratory, individuals were held overnight in this water with constant aeration. The
following day, fish were transferred into holding tanks (75 to 114 L aquaria) maintained at
the salinity at which fish were captured, and treated prophylactically for 7-14 days in a 5 mg
L'l solution of Acrifiavine. Following treatment, groups of approximately 10 fish were
placed into experimental (38 L aquaria) tanks containing water at a salinity within 10 ppt of
that in which they were collected. Both holding and experimental tanks were equipped with
undergravel filtration and constant aeration, and were maintained in rooms on a 12:12
lightrdark cycle. Fish were fed Tetramin flake food once each day. All experimental
aquaria were located in a constant temperature environment room that maintained aquaria at
20 1 C.
Experimental aquaria were used to acclimate fish to salinities ranging from 0 ppt to
100 ppt (0 to 2860 mOsm kg*l). The initial acclimation period was 14 days, after which
fish were either used in a metabolic trial or were transferred to the next higher or lower
salinity in the series. Salinity changes were in steps of 5 ppt, with smaller increments used
to acclimate fish to 0 ppt. This procedure was repeated until determinations had been made
at all experimental salinities. Water used in the freshwater acclimation was obtained from
wells in Alachua and Levy counties, Florida (mean conductivity = 360 pS cml).
Experimental salinities greater than freshwater but less than full seawater were prepared by
diluting filtered Atlantic Ocean seawater (obtained from the C. V. Whitney Laboratory of the
University of Florida, Marincland, Florida) with appropriate quantities of deionized water.
Salinities greater than 35 ppt were produced by supplementing seawater with appropriate


15
amounts of synthetic sea salt (Instant Ocean). Salinities were monitored daily with an
AO temperature-compensated refractometer, and adjusted as necessary.
Metabolic determinations were carried out in sealed, opaque flasks ranging in
volume from 0.6 to 1.175 L, with the volume selected based on the size of the fish being
tested. Rate of oxygen consumption was used to measure metabolism, with flasks being
used as closed respirometers. Measures of metabolism were considered to be routine
metabolic rates, as animals were sequestered in such a way as to minimize, but not eliminate
activity (Winberg, 1956; Fry, 1957). Experiments were performed with post-absorptive fish
in a resting state, but fish were unconstrained and capable of spontaneous motor activity
(Winberg, 1956; Fry, 1957). For a relatively inactive species such as C. variegatus
(compared to actively swimming salmonids, for example), RMR is perhaps the most
appropriate measure of metabolism, as it more closely reflects normal activity patterns of the
fish than does either active or standard metabolic rate.
Each flask was sealed with a rubber stopper through which two hypodermic needles
(No. 18) were inserted. Each needle was fitted on the inside with catheter tubing, one with a
piece long enough to reach the bottom of the flask, the other half this length. A 25 ml plastic
syringe filled with water at the salinity of the experimental aquarium was inserted into the
needle fitted with the short piece of catheter tubing, while an empty 10 ml syringe was
inserted into the other needle. A 1 ml water sample was drawn into the empty syringe for
each determination of PC>2- As each water sample was withdrawn, an equal volume of water
was injected from the filled syringe into the flask. This method of sampling, along with
minor movements by the fish, effectively stirred the water in the flask (Nordlie, pers.
comm.). Determinations of PO2 were made with a Radiometer oxygen electrode
connected to a Radiometer PHM 71 acid-base analyzer.
Measurements of the rate of reduction in PO2 were made at 0.5 h intervals, and
continued until fish had depleted the oxygen level to approximately 20 mm Hg (generally 5
to 9 h). Following the final PO2 determination, each fish was removed from its flask, damp-


16
dried and weighed to the nearest 0.01 g. All metabolic determinations were made between
0700 and 1900 hours, and fish were not re-used in other metabolic trials.
To ensure that fish were post-absorptive at the time of testing, food was withheld
from experimental aquaria for 24 h prior to beginning a metabolic reading. Respirometers
were filled with water at the salinity of the experimental aquaria, and placed in a water bath
maintained at 20 1 C. The entire metabolic apparatus was located in a small, semi-
darkened room in which no other activity took place. In order to allow time for the fish to
adjust to the respirometers, individuals were placed into the flasks (with constant aeration)
12 to 16 h before beginning a trial. At the beginning of the metabolic trial, aerators were
removed and the flask was sealed.
Calculation of oxygen saturation values for the experimental conditions (taking into
account salinity, temperature, relative humidity, and barometric pressure) were made using
the equations of Truesdale et al., 1955 and both RMR (mg 02 hl) and Pc (mm Hg) were
calculated for each fish. Data used for calculation of metabolic rates were limited to values
obtained while the POo in the respirometer was greater than 100 mm Hg, in order to ensure
that these calculations were made at oxygen tensions well above the Pc. All data were used
for calculation of the Pc. Determination of the Pc was made using a BASIC program to
calculate the critical point (Yeager and Ultsch, 1989). Following recommendations by
Yeager and Ultsch (1989), data for each fish were first plotted to ensure that the relationship
was a two-step function, following which the midpoint approximation was used to calculate
the Pc.
Oxygen consumption is strongly influenced by body mass, so RMR values were
mass-adjusted using an analysis of covariance (ANCOVA). Log mass-independent RMR
was used as the dependent variable and log mass as the covariate. Least square means
derived from the ANCOVA were used as adjusted RMR values. It was not possible to
perform an ANCOVA for the Pc values, so calculations of Pc were mass-corrected to the
value of the average mass (3.13 g) of all individuals used in this study. The exponent


17
describing the relationship between mass and metabolism for C. variegatus (Nordlie et al.,
1991), MR = kW0-68, was used to correct oxygen consumption rates. Values were corrected
following the relationship MRc = (W0a32)(3.13--32)(MRo), where MRc is the mass-
corrected oxygen consumption, W0 is the observed mass, and MRo is the observed oxygen
consumption at mass WQ (Ultsch et al., 1978; Cech, 1990). Statistical analyses follow
procedures outlined in Winer et al., (1991) and Sokal and Rohlf (1995). All statistical
analyses were one way tests using the Tukey-Kramer post hoc comparison (p = 0.05), and
values are given throughout as means standard error of the mean (se).
Field Measurements
Salt marsh habitats are widely considered to experience unpredictable and
fluctuating abiotic conditions. However, actual physicochemical measurements are
infrequently reported. To address this issue, field measurements were made at four sites in
the Cedar Key area over a one year period. Whenever possible, measurements at each site
were taken both at the surface and on the bottom (generally 1-1.5 m deep). Dissolved
oxygen, salinity, and temperature were measured one to three times each month between
0700 h and 1700 h, for a total of 19 dates between June 1990 and June 1991. Sites 1, 2, and
3 were located deep in the salt marsh where C. variegatus was routinely collected. These
sites were located in close proximity to one another (< 10 m apart), and were interconnected.
Unlike many locations in the salt marsh, these sites were never completely isolated from one
another or from connections to the Gulf of Mexico, even during the lowest tides. Site 4 was
located directly on the Gulf of Mexico, in the town of Cedar Key, approximately five km
from sites 1, 2, and 3. Although C. variegatus is present at site 4, collections were not made
at this location. These field measurements were not intended to indicate the complete ranges
of oxygen, salinity, or temperature experienced by organisms living within the salt marsh.
Thus, the actual ranges of physicochemical conditions experienced by C. variegatus are
most likely greater than reported here. However, these values arc a subset of the conditions


18
experienced by salt marsh inhabitants, and give an indication of some of the variability in
the measured physicochemical parameters.
Results
Routine Metabolism
Mean RMR was calculated for each fish and organized by salinity groups. Mean
values for unadjusted and adjusted RMR (from ANCOVA) are given in Table 2-1, and
adjusted RMR are plotted against ambient salinity in Figure 2-1.
In the range of ambient salinities between freshwater and 40 ppt, adjusted RMR
values were highest at 2 ppt and 40 ppt, being slightly lower and roughly equivalent at the
other measured salinities in this range. At salinities greater than 40 ppt, there was a
progressive decline in adjusted RMR. Overall, adjusted RMR ranged from a maximum of
0.97 mg 02 hl at 2 ppt, to a low of 0.64 mg 02 h'l at 100 ppt, representing a 66%
decline. This decline corresponds with a decreased ability to regulate plasma osmolality at
elevated salinities (Figure 2-2; plasma osmolality data are from Nordlie, 1985).
A multiple linear regression analysis was used to generate a predictive model for
describing the effects of both salinity and mass on metabolism. In this model, Log body
mass (Log W; in g) and salinity (S; in ppt) were used as independent variables, and Log
mass-independent RMR (Log MR; in mg O2 h'l) as the dependent variable. The equation
that best described this relationship is
Log MR = -0.296 + 0.548 (Log W) 0.001 (S) (Fo.l 10 = 85.445; P < 0.0001)
This model described 62% of all variability about the mean, and the random distribution of
the residuals suggests an absence of significant relationships that might have biased the
analysis.


Table 2-1. Relationships of routine metabolism (RMR), critical oxygen tension (Pc), and slope in the conformation region at a
series of ambient salinities. Values arc given as means se.
Ambient Salinity
(PPO
n
Mean Body
Mass
(g)
Unadjusted RMR
(mg C>2 h'l)
Adjusted RMR
(mg 02 h'1)
Critical Oxygen
Tension
(mm Hg)
Slope in
Conformation
Region
0
11
3.17 0.232
0.81 0.038
0.78 0.025
56.98 6.92
0.0071 0.0024
2
7
3.38 0.258
1.04 0.017
0.97 0.027
51.49 5.85
0.0079 0.0039
15
8
2.95 0.438
0.75 0.048
0.78 0.029
53.68 4.92
0.0072 0.0021
30
10
3.57 0.402
0.88 0.033
0.83 0.020
52.16 5.10
0.0058 0.0018
40
9
2.19 0.337
0.76 0.050
0.96 0.030
52.14 2.70
0.0079 0.0017
50
18
3.69 0.322
0.94 0.031
0.87 0.020
61.81 4.89
0.0065 0.0011
60
8
3.42 0.491
0.83 0.057
0.80 0.030
63.41 6.06
0.0033 0.0008
70
12
2.70 0.389
0.65 0.050
0.73 0.025
66.31 5.57
0.0041 0.0006
80
12
2.98 0.437
0.67 0.047
0.70 0.025
79.53 5.61
0.0033 0.0004
90
8
3.52 0.362
0.73 0.040
0.68 0.029
74.94 8.00
0.0034 0.0004
100
8
3.21 0.460
0.63 0.032
0.64 0.030
73.93 8.78
0.0034 0.0005


Figure 2-1. Mean adjusted routine metabolic rates (RMR) over a range of salinities in Cyprinodon vciriegatus (metabolic rates were mass-
adjusted using an analysis of covariance; bars indicate se; numerical values above the points in the figure indicate sample size at each
salinity).


Salinity (ppt)
Adjusted Routine Metabolic Rate (mg Oxygen hr )
poop f- r
i*. b\ bo ^ i*.
\Z


Figure 2-2. Relationship between mean adjusted routine metabolic rates (RMR) and mean plasma osmolality over a range of salinities in
Cyprinodon variegatus (metabolic rates were mass-adjusted using an analysis of covariance; bars indicate se; plasma osmolality data
from Nordlie, 1985).


Adjusted Routine Metabolic Rate
(mg Oxygen/hr)
Plasma Osmolality (mOsm/kg)


24
Critical Oxygen Tension
Measurements of oxygen consumption (mg 02 h'l) for each fish at all time
intervals were mass-adjusted to the mean mass of all individuals used in this study. These
mass-adjusted values were then used to calculate the Pc for each fish. Mean values of Pc
were organized by salinity group and are given in Table 2-1. Plots showing the relationship
between Pc and ambient salinity and between Pc, RMR, and ambient salinity are shown in
Figures 2-3 and 2-4, respectively.
Mean Pc values in the range of ambient salinities from 0 ppt to 40 ppt were not
significantly different from one another (p = 0.95), similar to the pattern exhibited by the
RMR data Mean Pc values increased at salinities greater than 40 ppt, with the highest levels
recorded at salinities 80 ppt and higher. Pc values ranged from a low of 51.49 mm Hg at a
salinity of 2 ppt, to a high of 79.50 mm Hg at 80 ppt, representing a 45% increase. The rise
in mean Pc values corresponds well with a decreased ability to regulate plasma osmolality,
again similar to the RMR pattern (Figure 2-5; plasma osmolality data are from Nordlie,
1985).
Below the Pc, metabolism depends on the oxygen tension and decreases as the PO2
decreases. Mathematically, the slope of the resulting line in this conformation region
depends on three factors: the Pc, the RMR at the Pc, and the lethal P02- A comparison of
the slopes in the conformation region reveals that despite the changes noted above in RMR
and Pc values, the PO2 at which the fish can no longer survive (under experimental
conditions) is essentially equivalent for all salinities tested. This is reflected in the
increasingly shallow slopes seen at salinities greater than 50 ppt (Table 2-1).
Field Measurements
The field measurements of oxygen concentration, salinity, and temperature revealed
very high variability of these physicochemical parameters over the course of the sampling


Figure 2-3. Mean critical oxygen tension (Pc) measurements over a range of salinities in Cyprinodon variegcitus (bars indicate se
numerical values above the points in the figure indicate sample size at each salinity).


Salinity (ppt)
Critical Oxygen Tension (mm Hg)
u tn o\ oo vo
o o o o o o
9c
100


Figure 2-4. Relationship between mean adjusted routine metabolic rates (RMR) and critical oxygen tensions (Pc) over a range of salinities
in Cyprinodon variegatus (metabolic rates were mass-adjusted using an analysis of covariance; bars indicate se).


Adjusted Routine Metabolic Rate
(mg Oxygen/hr)
0 10 20 30 40 50 60 70 80 90 100 110
Salinity (ppt)
Critical Oxygen Tension (mm Hg)


Figure 2-5. Relationship between mean critical oxygen tensions (Pc) and mean plasma osmolality over a range of salinities in Cyprinodon
variegatus (bars indicate se; plasma osmolality data from Nordlie, 1985).


Critical Oxygen Tension (mm Hg)
100
700
90 -
Critical Oxygen Tension
Plasma Osmolality
80 -
70 -
60 -
50 -
40 -\ 1 1 1 1 1 1 | i 1 | 1 , r
0 10 20 30 40 50 60 70 80 90 100
Salinity (ppt)
- 650
- 600
- 550
- 500
- 450
- 400
- 350
-- 300
110
Plasma Osmolality (mOsm/kg)


31
period (Table 2-2). For all sites combined, ranges of dissolved oxygen, salinity, and
temperature over the course of the year were 0.6 10.8 mg L'l, 1.0 29.0 ppt, and 9.0 -
38.0 C, respectively. Even higher salinities (> 40 ppt) were encountered occasionally in the
salt marsh outside the sampling period. Other areas of the salt marsh probably encounter
even greater extremes of these variables. Cyprinodon variegatus was seen on all dates when
physicochemical measurements were made.
In addition to high overall variability, there were large differences among some sites
in close proximity to one another. For example, mean oxygen concentration measurements
taken during the Fall at sites one, two, and three were 2.94 mg L 1, 3.85 mg L" 1, and 5.1 mg
L'l, respectively. All three physicochemical parameters also strongly varied temporally
among sampling dates. These data provide good evidence that the Cedar Key salt marsh is
an extremely variable habitat with respect to these physicochemical parameters.
Discussion
The family Cyprinodontidae is a diverse group of fishes with many species that
tolerate extreme environmental conditions (Lowe et al., 1967; Lotan and Skadhauge, 1972;
Naiman et al., 1976; Stuenkel and Hillyard, 1981; Chung, 1982). Cyprinodon variegatus is
perhaps the most physiologically tolerant member of the family. It has been called "the
toughest fish in North America" (Gunter, 1967) due to its extreme tolerance of severe
abiotic conditions. The species is found in waters ranging in salinity from freshwater (Ager,
1971) to 142 ppt (Simpson and Gunter, 1956), and can reproduce in waters as high as 100
ppt (Martin, 1972). They are known to tolerate temperatures ranging from about 1 C
(Berry, 1987), to temperatures greater than 41 C (Strawn and Dunn, 1967), and to tolerate
near anoxic conditions (Odum and Caldwell, 1955). Thus, this species is an exceedingly
useful experimental subject for examining how tclcosts respond to harsh environmental
conditions.


Table 2-2. Measurements of oxygen concentration (mg L'l), salinity (ppt), and temperature (C) taken at four sites in the Cedar Key
area from June 1990 through June 1991 (see text for details on location of sites). Values are given as means, se (in parentheses),
sample size.
Oxygen (mg L"l)
Salinity (ppt)
Temperature (C)
Location
Depth
Spring
Summer
Fall
Winter
Spring
Summer
Fall
Winte
r
Spring
Summer
Fall
Winter
Site 1
2.18
3.15
2.63
4.24
13.9
16.63
12.75
11.3
25.2
30.50
21.75
14.90
Bottom
(0.28)
(0.54)
(0.42)
(0.97)
(1.05)
(1.45)
(2.62)
(1.69)
(0.37)
(1.03)
(2.39)
(1.35)
n = 5
n = 8
n = 4
n = 5
n = 5
n = 8
n = 4
n = 5
n = 5
n = 8
n = 4
n = 5
3.08
3.72
3.25
5.16
11.00
15.92
13.75
10.60
25.60
31.83
21.63
15.00
Surface
(0.38)
(0.60)
(0.51)
(136)
(1.04)
(2.88)
(2.44)
(0.80)
(1.37)
(1.49)
(2.93)
(1.18)
n = 5
n = 6
n = 4
n = 5
n = 5
n = 6
n = 4
n = 5
n = 5
n = 6
n = 4
n = 5
Site 2
1.98
2.37
2.65
6.78
12.30
13.56
12.50
7.00
24.8
29.36
20.00
14.40
Bottom
(0.47)
(0.70)
(0.63)
(1.33)
(2.65)
(2.31)
(2.73)
(0.91)
(0.84)
(0.69)
(2.81)
(1.54)
n = 5
n = 8
n = 4
n = 5
n = 5
n = 8
n = 4
n = 5
n = 5
n = 8
n = 4
n = 5
3.35
4.83
5.05
8.56
6.2
11.08
7.50
4.20
24.70
30.42
20.13
14.60
Surface
(0.36)
(0.56)
(0.89)
(0.66)
(1.47)
(3.29)
(2.60)
(0.72)
(0.97)
(0.84)
(3.18)
(1.63)
n = 5
n = 6
n = 4
n = 5
n = 5
n = 6
n = 4
n = 5
n = 5
n = 6
n = 4
n = 5
Site 3
3.83
3.46
5.08
8.24
6.80
7.20
7.88
4.60
24.90
30.40
21.38
16.60
Bottom
(0.43)
(1.02)
(0.96)
(0.48)
(1.97)
(2.96)
(3.18)
(0.78)
(0.78)
(1.29)
(3.18)
(1.29)
n = 5
n = 5
n = 4
n = 5
n = 5
n = 5
n = 4
n = 5
n = 5
n = 5
n = 4
n = 5
3.65
5.03
5.15

4.50
5.33
3.00

25.30
31.17
25.50

Surface
(0.25)
(0.73)
(0.15)

(0.50)
(1.92)
(2.00)

(1.33)
(1.59)
(0.5)

n = 3
n = 3
n = 2

n = 3
n = 3
n = 2

n = 3
n = 3
n = 2

Site 4
5.10
5.37
4.35
8.06
17.75
27.36
23.88
15.50
27.0
29.71
22.50
15.10
Middle
(0.68)
(0.85)
(0.60)
(0.98)
(3.13)
(0.38)
(0.52)
(2.42)
(0.51)
(0.38)
(2.76)
(1.36)
n = 5
n = 7
n = 4
n = 5
n = 5
n = 7
n = 4
n = 5
n = 5
n = 7
n = 4
n = 5


33
Field measurements indicated that the Cedar Key salt marsh varies tremendously,
both spatially and temporally, in dissolved oxygen, temperature, and salinity, thus exposing
organisms living within the salt marsh to extreme environmental conditions. Cyprinodon
variegatus tolerates these conditions very well. Salinities between 0 ppt and 40 ppt have
little effect on the energetics of C. variegatus as measured by RMR and Pc- This species
fits into the Typ>e I metabolic response pattern (species exhibiting no significant change in
metabolic rate over a wide range of environmental salinities) as categorized by both Nordlie
(1978) and Morgan and Iwama (1991). A similar pattern characterizes the related salt marsh
resident, Adinia xenica (D.C. Haney, unpublished data), in which RMR was more variable,
but relatively constant over salinities from 0 ppt to 50 ppt (Figure 2-6).
Cyprinodon variegatus exhibits a decline in RMR only at salinities exceeding 40
ppt. This result was somewhat unexpected, but is nearly identical to the pattern found by
Nordlie et al., (1991) in a study that also examined metabolism of C. variegatus over a wide
range of ambient salinities. Nordlie et al., (1991) concluded that the depression in
metabolism at high salinities is probably related to permeability changes of the gill
membrane and/or integument. The point at which metabolism is reduced corresponds well
with a diminished ability of C. variegatus to osmoregulate efficiently. If osmotic
permeability of the gills is reduced at high salinities to help offset ionic influx and osmotic
efflux, the potential for oxygen uptake may be reduced as well (Kristensen and Skadhauge,
1974; Skadhauge, 1974; Davenport and Saycr, 1993). Evidence for this hypothesis comes
from several recent studies. In the first of these, Kultz and Onken (1993) found that overall
in vitro permeability of the opercular membrane of the cichlid Oreochromis mossambicus
was reduced in hypcrsaline media, with a simultaneous reduction in passive ion fluxes.
More direct evidence comes from studies by Bindon et al., (1994a) and Bindon et al.,
(1994b), on the rainbow trout, Oncorhynchus mykiss. In these studies the authors
demonstrated that impairment of respiratory gas transfer coincided with chloride cell
proliferation induced by an osmoregulatory challenge.


Figure 2-6. Relationship between mean adjusted routine metabolic rates (RMR) and mean plasma osmolality over a range of salinities in
Adinia xenica (metabolic rates were mass-adjusted using an analysis of covariance; bars indicate se; numerical values above the points
in the figure indicate sample size at each salinity).


Adjusted Routine Metabolic Rate
(Mg Oxygen/hr)
600
Salinity (ppt)
Plasma Osmolality (mOsm/Kg)


36
Changes in activity may also account for a depression in metabolism. In a study on
the milkfish, Chanos chanos, Swanson (1991) showed that a depression in metabolism at
an elevated salinity (55 ppt) was highly correlated with a reduction in activity. This may
have been a factor in this study although the method used here for metabolic determinations
minimized activity of fish at all salinities.
The main focus of this study was determination of Pc over a wide salinity range, as
no other study has investigated to any extent the influence of salinity on this parameter.
Critical oxygen tension can be affected by many factors. These include both physical and
biotic parameters such as temperature, salinity, size and activity, reproductive and nutritional
state, and experimental parameters, such as rate of oxygen depletion or manipulation and
disturbance of fish during measurements (McMahon, 1988).
Like other vertebrates, C. variegatus is a good oxygen regulator over a wide range
of oxygen tensions. Critical oxygen tension in C. variegatus was unaffected by changes in
salinity between 0 ppt and 40 ppt, similar to the result for RMR. However, above 40 ppt, Pc
increased with further increases in salinity. This seems reasonable, as fish might be
expected to encounter difficulties obtaining sufficient oxygen at elevated salinities.
Unfortunately, it is difficult to compare my results with those from other groups of
fishes. While the Pc has been measured for a number of freshwater, marine, and intertidal
fishes, only a few authors have considered the effects of salinity in their analyses. Job
(1969) examined oxygen consumption of Tilapia mossambica (= O. mossambicus) at 0.4
ppt, 12.5 ppt, and 30.5 ppt, and found no effect of salinity on the Pc of small (5 g) fish at
either 15 C, 30 C, or 40 C (Pc approximately 50 mm Hg). Salinity did affect Pc on
larger (80 g) individuals at 15 C, where the Pc doubled from approximately 50 mm Hg to
100 mm Hg in fish acclimated to 0.4 ppt versus 12.5 ppt or 30.5 ppt. However, Pc
calculations made by Job (1969) were extremely rough approximations, and are difficult to
compare with values in this study.


37
Subrahmanyam (1980) examined the influence of oxygen tension on the metabolic
rate of several salt marsh fishes including C. variegatus, with all measurements made at 25
C and salinities of 17-21 ppt. All species tested in Subrahmanyam (1980) (C. variegatus,
Poecilia latipinna, Lagodon rhomboides, Leiostornus xanthurus, Fundulus granis, and
F. similis) were oxygen conformers at oxygen tensions of 85 mm Hg and lower, a
somewhat different response than seen in this study. This difference may be due to the
small sample sizes, and relatively few measurements of oxygen consumption made by
Subrahmanyam (1980). In a study on the golden mullet, Liza aurata, Shusmin (1989),
measured the "oxygen threshold" at salinities ranging from freshwater to 50 ppt. He
showed that the oxygen threshold was relatively constant at all salinities between freshwater
and 40 ppt, with a large increase at 50 ppt. Cyprinodon variegatus did not show this type of
response, as the lethal endpoint does not appear to increase at higher salinities in this
species.
Values of the Pc in this study compare fairly well with Pc measurements for other
groups of fishes. Intertidal marine species generally have lower Pc values than C.
variegatus, ranging from 20 to 26 mm Hg in Paraclinus intergripinnis (Congleton, 1980)
to 30 to 40 mm Hg in Gobius cobitus (Bridges, 1988) and Helcogramma medium (Innes
and Wells, 1985; Pelster et al., 1988; Quinn and Schneider, 1991). A number of African
cichlids ( e.g., Oreochromis niloticus, Cichlasoma urophlhalamus, Erelmodus
cyanosticus, Dimidiochromis compressiceps) have either slightly lower, or roughly
equivalent Pc values to the approximate value (55 mm Hg) displayed by C. variegatus in
this study at most salinities (Ross and Ross, 1983; Becker and Fishelson, 1986; Palacios
and Ross, 1986; Verhcycn ct al., 1994). Donnelly and Torres (1988) found Pc values
ranging from 25 to 50 mm Hg for a number of midwatcr fishes from the eastern Gulf of
Mexico, again values near, or slightly lower than those of C. variegatus. Thus, C.
variegatus has Pc values that arc fairly close to those of ecologically and evolutionarily
diverse tclcosts.


38
The variations in RMR and Pc as a function of environmental salinity observed in
this study suggest that C. variegatus responds to high salinities by reducing energy
expenditures. Observations by myself and others indicate that hypersaline conditions are
encountered infrequently, and likely last for short (days) periods of time. Available evidence
suggests that high salinities are metabolically expensive, whether due to an influence on
activity, osmoregulation, or other physiological and/or behavioral processes. Low oxygen
conditions also commonly occur with high salinities. Decreased metabolism in conjunction
with increased Pc reduces energetic expenditures dramatically at elevated salinities. These
responses effectively increase the time C. variegatus can tolerate adverse conditions, albeit
at a cost of a reduction in energetic processes. However, this is an appropriate response to
harsh conditions that appear at variable and unpredictable time intervals, but that are present
for only short periods of time.
The type of metabolic response to salinity in C. variegatus fits the concept of
"scope for survival" as described by Hochachka (1990). This is a pattern of metabolic
response to environmental stressors characterized by a depression in metabolism,
sometimes below maintenance levels. The advantage of such a reduction in energy
expenditure is essentially the slowing of biological time, enabling survival despite the
temporarily imposed physiological stressor.
Cyprinodon variegatus is an extremely successful euryhaline fish that can survive a
large range of salinities, with typically encountered salinities having little effect on normal
metabolism or on the ability of individuals to osmoregulate efficiently. Fish accomodate
extremely high salinities by reducing energetic expenditures. Such a response increases the
amount of time shcepshead minnows can survive hypcrsaline conditions, enabling them to
"wait out" the difficult conditions.


CHAPTER 3
INFLUENCE OF SIMULATED TIDAL CHANGES IN AMBIENT SALINITY ON
ROUTINE METABOLIC RATE IN CYPRINODON VARIEGATUS
Introduction
Salt marshes often undergo large and rapid salinity fluctuations, a condition that
may significantly affect the distribution and abundance of organisms within these habitats.
Changes in salinity may dramatically influence the energetics of individual fish. Information
on the metabolic costs associated with salinity fluctuations may be useful in explaining or
predicting distribution patterns of some coastal fish species.
Energetic patterns of fishes have been examined relative to a number of abiotic
factors, most notably temperature and oxygen (e.g., Wells, 1935; Fry, 1957; Beamish, 1964;
Brett and Groves, 1979; Stuenkel and Hillyard, 1981; Johnston and Battram, 1993).
Comparatively fewer studies have been considered the influence of salinity on metabolic
patterns of fishes (e.g., Kinne, 1966; Madan Mohon Rao, 1974; Nordlie et al., 1991;
Swanson, 1991). In these studies, various metabolic responses to salinity have been
reported. Euryhaline fish exhibit one of the few consistent patterns, namely that metabolism
remains relatively unaffected over the range of salinities normally encountered (Nordlie,
1978; Morgan and Iwama, 1991).
Most studies that have examined the influence of salinity on fish energetics have
employed measurements from fish maintained at constant salinities. Data from these
experiments are very useful for understanding the overall metabolic patterns exhibited by
the species tested, as fish were usually fully acclimated to each experimental salinity prior to
testing. However, these experiments provide less information about more ecologically
39


40
relevant responses, as fish in their native habitats are often subject to rapid and dramatic
fluctuations in salinity.
Studies of the influence of salinity fluctuations on metabolism of fishes have
usually focused on species that generally experience maximal fluctuations in salinity only
between freshwater (0 ppt) and seawater (35 ppt) (e.g., Davenport and Vahl, 1979; Von
Oertzen, 1984; Moser and Gerry, 1989; Shusmin, 1989; Moser and Miller, 1994).
However, some salt marsh teleosts regularly encounter salinities outside this range. This
study examined the influence of salinity fluxes on routine metabolic rate (RMR) of the salt
marsh teleost, Cyprinodon variegatus, a species that regularly encounters salinities greater
than 35 ppt.
Fish used in this study were fully acclimated to a series of salinities ranging from 0
ppt to 60 ppt, followed by exposure to a simulated tidal change in salinity. The magnitude,
rate, and direction of salinity changes may be important determinants of how salinity
fluctuations affect metabolism. In this study, the direction of the salinity change in
conjunction with the ambient (acclimation) salinity was the primary focus. The magnitude
of the salinity changes were selected to simulate extremes known to occur in the habitat
from which fish were collected, and the rate was chosen to simulate a normal diurnal tidal
cycle. I predicted that C. variegatus would show minimal changes in RMR when the
salinity was changed within the range commonly encountered by the species. Salinity
changes outside this range were hypothesized to lead to depressions in metabolism.
Methods
Collections of fish, transportation back to the laboratory, and general laboratory
procedures were performed as described previously. Fish were first sequentially acclimated
to a series of salinities ranging from 0 ppt to 60 ppt (0 to 1715 mOsm kg'l). However,
unlike the previous experiments, these procedures were designed to measure RMR before


41
and following simulated tidal changes in salinity. Initial salinities (acclimation) and salinities
following the simulated tidal change (final) are shown in Table 3-1.
Metabolic determinations and changes in salinity were carried out in a flow-through
respirometer with an effective volume of 0.875 L (Figure 3-1). The respirometer was
immersed in a thermoregulated reservoir during metabolic trials that served to maintain a
constant temperature within the respirometer. A submersible pump maintained within a
second thermoregulated reservoir (38 L) was connected to the inlet of the respirometer. This
reservoir was vigorously aerated and was used to supply water to the respirometer. The
respirometer outlet emptied back into this main reservoir. A third reservoir (19 L) serv ing as
a salinity source was connected to the main reservoir via a peristaltic pump. Due to
limitations of the available equipment, the system was closed during oxygen partial pressure
(PO2) determinations. Two ports within the respirometer were used to sample water for
determination of P02- A 10 ml plastic syringe filled with water at the experimental salinity
was inserted into the first port (inlet side), while an empty 10 ml syringe was fitted into the
second port (outlet side). As a 1 ml water sample was drawn into the empty syringe, an
equal volume of water was injected from the filled syringe into the respirometer.
Determinations of PO2 were made with a Radiometer oxygen electrode connected to a
Radiometer PHM171 acid-base analyzer.
To begin a metabolic trial, the main reservoir and respirometer were filled with water
at the acclimation salinity. Fish were placed into the respirometer 12 h prior to the
beginning of a metabolic determination. This allowed fish sufficient time to adjust to the
experimental setup. The respirometer was then sealed and immersed in the thermoregulation
reservoir, and the submersible pump was turned on at a flow rate of 1 L min'*. The
following morning, the pump was turned off, the system was closed, and PO2
measurements begun. Measurements of the rate of reduction in PO2 were made at 0.5 to
1.0 h intervals, and continued until fish had depleted the oxygen level to approximately 100
mm Hg (generally 4 to 6 h). Following the final PO2 determination, the system was


42
Table 3-1. Acclimation and final salinities used in simulated tidal change study.
Acclimation Salinity
Final Salinity
*
Oppt
0 ppt (control)
20 ppt
2 ppt
2 ppt (control)
20 ppt
30 ppt
10 ppt
10 ppt (control)
30 ppt
20 ppt
Oppt
2 ppt
20 ppt (control)
40 ppt
50 ppt
30 ppt
2 ppt
10 ppt
30 ppt (control)
50 ppt
40 ppt
20 ppt
40 ppt (control)
60 ppt
50 ppt
20 ppt
30 ppt
50 ppt (control)
60 ppt
40 ppt
60 ppt (control)
reopened, and the submersible pump turned on. At this time, either well water (0 ppt) or a
saline solution of variable concentration was placed into the salinity reservoir. The salt
concentration in this reservoir was determined so that its addition to the main reservoir
would change the salinity of the main reservoir and respirometer to the desired final salinity.


Figure 3-1. Schematic diagram of respirometry apparatus used for routine metabolism experiments. See text for detailed description of
system.




45
The volumes of both reservoirs and the rate of pumping by the peristaltic pump were
adjusted so that the salinity would change at a uniform rate over a 6 h period. The peristaltic
pump was plugged into an automatic timer so that salinity changes took place between 1900
h to 0200 h.
On the second morning the system was re-closed and PO2 measurements were
made at the new (final) salinity. PO2 measurements at the final salinity were thus made 6-
12 h following completion of the salinity fluctuation, a period sufficiently long to be past
the stressful transition period (Von Oertzen, 1984). Following the final P02 determination,
the fish was removed from the respirometer, damp-dried and weighed to the nearest 0.01 g.
All metabolic determinations were made between 0700 h and 1500 h, and fish were not re
used in other metabolic trials. Fish were thus held within the respirometer for a total of 40
to 44 h (including adjustment period).
Food was withheld from experimental aquaria for 24 h prior to beginning a
metabolic reading to ensure that fish were post-absorptive at the time of testing. Fish were
not fed while in the respirometer. The entire metabolic apparatus was located in a small,
semi-darkened room in which there was no other activity, with determination of metabolic
rates performed as previously described.
Because a metabolic determination for the two salinity treatments was performed on
the same individual, it was not necessary to mass-correct values in order to perform the
statistical analyses. However, to compare data obtained in this study more easily with
published literature, all RMR measurements were mass-adjusted to the value of the average
mass (2.94 g) of all individuals used in this study. Adjusted RMR values were calculated as
previously described. A repeated measures analysis of variance was used to compare
adjusted RMR's between acclimation and final salinities. Statistical analyses follow
procedures outlined in Winer etal. (1991) and Sokal and Rohlf (1995). All analyses of
variance were one way tests using the Tukey-Kramcr post hoc comparison (p = 0.05).


46
Results
Comparison of RMR's at acclimation and final salinities revealed some interesting
patterns (Table 3-2). There were no significant differences (p > 0.05) in RMR between
acclimation and final salinities in any control trials (n = 4 each). Similarly, when both
acclimation and final salinities (2 ppt, 10 ppt, 20 ppt, 30 ppt, and 40 ppt) were in a range that
is typically encountered by C. variegatus, there was a significant change in RMR in only
one trial. Typical salinities in the wild range from 2-5 ppt through 30-35 ppt, with
hypersaline conditions (35-40 ppt) occurring much more frequently than salinities near 0
ppt (pers. obs.). When the acclimation salinity is high (50 ppt and 60 ppt), all groups
exhibited significant elevation of RMR in the lower, more typical, final salinity. The same
general pattern was seen when the final salinity was high, where fish in two of three groups
had depressed RMR at the highest salinities. RMR was depressed when either the
acclimation or final salinity was 0 ppt. The direction of salinity change strongly influenced
the metabolic response. When salinity was increased over the course of the trial (Figure 3-
2), fish were only affected metabolically at the very highest and lowest salinities, where
metabolism was depressed. When salinity was decreased over the course of the trial (Figure
3-3), fish again showed metabolic depression at extreme salinities, but, overall, more groups
exhibited changes in metabolism with changes in salinity than groups in which salinity was
increased.
Discussion
My results demonstrate that C. variegatus maintains a very stable metabolism when
exposed to typical salinity fluctuations seen in the salt marsh habitat at Cedar Key, Florida.
The experimental procedure resulted in no mortalities during or immediately following any
salinity trial. Metabolism was generally unaffected in salinity trials where both acclimation
and final salinities were in the range typically encountered by C. variegatus at Cedar Key. It


47
Table 3-2. Mean routine metabolism (mg Ch h'l) before (acclimation salinity) and
following (final salinity) a simulated tidal change. Values are given as means se. Groups
exhibiting a significant change in metabolism are indicated with an asterisk.
Acclimation
Salinity
(PPO
Acclimation
RMR
(mg 02 h'l)
Final
Salinity
(PPQ
Final
RMR
(mg C>2 h'l)
P value
Response
Qppt
0.466 0.07
20 ppt (n = 5)
0.584 0.09
0.04 *
Increase
2 ppt
1.242 0.18
20 ppt (n = 5)
1.244 0.23
0.99
No Change
1.024 0.13
30 ppt (n = 5)
0.978 0.06
0.69
No Change
10 ppt
1.084 0.20
30 ppt (n = 5)
1.082 0.19
0.97
No Change
20 ppt
1.348 0.14
0 ppt (n = 5)
0.906 0.09
0.01 *
Decrease
0.886 0.03
2 ppt (n = 5)
0.478 0.07
0.01 *
Decrease
0.948 0.08
40 ppt (n = 6)
0.785 0.17
0.33
No Change
1.025 0.11
50 ppt (n = 6)
0.745 0.07
0.01 *
Decrease
30 ppt
1.028 0.23
2 ppt (n = 5)
0.854 0.10
0.33
No Change
0.628 0.07
10 ppt (n = 5)
0.688 0.11
0.34
No Change
0.728 0.07
50 ppt (n = 6)
0.728 0.12
1.00
No Change
40 ppt
0.704 0.12
20 ppt (n = 5)
0.662 0.09
0.75
No Change
0.626 0.06
60 ppt (n = 5)
0.476 0.03
0.03 *
Decrease
50 ppt
0.620 0.04
20 ppt (n = 5)
0.904 0.11
0.05 *
Increase
0.850 0.12
30 ppt (n = 5)
1.076 0.14
0.05*
Increase
60 ppt
0.452 0.07
40 ppt (n = 5)
0.578 0.05
0.01 *
Increase
was only when the acclimation or final salinities were outside the range normally
encountered (i.c., 0 ppt, 50 ppt, and 60 ppt) that changes in metabolism were found. In these
cases metabolism was depressed at each extreme salinity relative to the measurements made
at the more typical salinity.


Figure 3-2. Results of metabolic trials where salinity was increased over the course of the
trial. Bars represent groups listed in Table 3-2 for which final salinity was greater than
initial salinity. The height of each bar signifies the magnitude of the salinity change for each
metabolic trial and the asterisk indicates at which of the salinities (for each metabolic trial)
the routine metabolic rate (RMR) was highest The x axis has no scale and serves only to
visually separate groups.


Salinity (ppt)
49
60
50
40
30
20
10
2
0
RMR's Not Significantly Different (p = 0.05)
RMR's Significantly Different (p = 0.05)
*
*
*
Salinity Increasing


Figure 3-3. Results of metabolic trials where salinity was decreased over the course of the
trial. Bars represent groups listed in Table 3-2 for which final salinity was less than initial
salinity. The height of each bar signifies the magnitude of the salinity change for each
metabolic trial and the asterisk indicates at which of the salinities (for each metabolic trial)
the routine metabolic rate (RMR) was highest. The x axis has no scale and serves only to
visually separate groups.


Salinity (ppt)
51
60
50
40
30
20
10
2
0
RMR's Not Significantly Different (p = 0.05)
RMR's Significantly Different (p = 0.05)
* *
*
Salinity Decreasing


52
These results allow for limited comparison because similar studies have examined
the influence of salinity over a much narrower range than examined in this study. The best
comparison is with a study by Wakeman and Wohlschlag (1983) on the red drum,
Sciaenops ocellatus. In their study, test animals were acclimated to a series of salinities
between 1 ppt and 50 ppt, and then abruptly transferred to salinities either 10 ppt higher or
lower than the acclimation salinity. Although Wakeman and Wohlschlag (1983) did not
specifically compare metabolic rates following the transfer with those taken at the
acclimation salinity, there were trends similar to results of this study. Fluctuations in salinity
over a moderate range caused little or no change in metabolism of S. ocellatus on
determinations made 12 to 24 h following the transfer. Only when either the acclimation or
final salinity was 50 ppt were changes in metabolism obvious, with metabolic rates elevated
in 50 ppt compared to 40 ppt. It thus appears that 5. ocellatus responds to abrupt salinity
changes somewhat differently than docs C. variegatus, as metabolism increased upon
transfer to 50 ppt in S. ocellatus, but decreased in C. variegatus. This difference may be
due to the smaller magnitude of the salinity changes performed by Wakeman and
Wohlschlag (1983) (10 ppt vs 20 to 30 ppt in the present study). However, these results
could also be attributable to different sizes and activity patterns of experimental fish
between the two studies.
In a similar study, Shushmin (1989) studied the dynamics of oxygen consumption
in juvenile golden mullet, Liza aurata, following abrupt transfers from 18 ppt to a series of
salinities ranging from 0.4 ppt to 50 ppt. All salinity changes resulted in increased
metabolic rates for one to three days following the transfer, with metabolism generally
returning to near normal levels within three to four days. It is not surprising that salinity
changes arc more stressful in the golden mullet, since the species has lower salinity
tolerance and osmoregulatory ability than C. variegatus.
Barton and Barton (1987) examined metabolism in juvenile (< 1 g) C. variegatus
collected from inland saline lakes of San Salvador Island. They measured metabolic rates of


53
fish at 10 ppt and 35 ppt following abrupt transfer from an acclimation salinity of 35 ppt.
Metabolic rates in fish measured at 10 ppt were significantly higher than those made at the
acclimation salinity. These results are somewhat different from those obtained in the present
study. However, measurements by Barton and Barton (1987) were made only 3 h following
abrupt transfer to the reduced salinity, whereas measurements in the present study were
made 6-12 h following a gradual change in salinity. Fish in this study thus experienced a
less stressful transition to the new salinity. An ontogenetic effect may also account for the
observed differences, as many investigations have demonstrated that adults and juveniles of
the same species may differ greatly in physiological competence (e.g., Kinne, 1966; Martin,
1968; Oikawa et al., 1991).
Other relevant comparisons can be made with data on juvenile croaker,
Micropogonias undulatus, and juvenile spot, Leiostomus xanthurus (Moser and Gerry,
1989; Moser and Miller, 1994). These studies examined effects on metabolism from
fluctuations in salinity between 0 ppt and 35 ppt. Like C. variegatus, a salinity decrease
from higher salinities to 0 ppt generally led to a decreased metabolism in L. xanthurus.
Similarly, when fish were acclimated to 0 ppt and salinity was increased, metabolism was
elevated at the higher salinities. Contrary to findings for C. variegatus, salinity fluctuations
between 15 ppt and 35 ppt elicited changes in metabolism in both L. xanthurus and M.
undulatus. Thus, these species are influenced to a greater extent by salinity fluctuations than
C. variegatus. However, rates of salinity fluctuation in the above studies were faster than
rates used in the present study (16 or 32 ppt h'l vs 3.3 or 5 ppt hl), and C. variegatus
likely experiences greater fluctuations in salinity than do spot or croaker.
Acclimation state is the most important measured factor influencing the metabolic
response of C. variegatus to simulated tidal changes in salinity. However, direction of the
salinity change also influenced metabolism in C. variegatus. Like the results obtained by
Moser and Miller (1994) on juvenile L. xanthurus, it appears that C. variegatus adjusts to
increasing salinity more effectively than to decreasing salinity, as evidenced by the


54
responses to salinity changes between 20 ppt and 2 ppt, and between 50 ppt and 30 ppt.
Thus, when salinity is increased over the course of the trial, fish are only affected
metabolically at the very' highest and lowest salinities. However, when fish are acclimated to
a high salinity, and salinity is then decreased to a more typical range, individuals take
advantage by increasing metabolism to more normal levels.
Thus, C. variegatus is well adapted to a varying salinity environment. Its
metabolism is unaffected by changes in salinity over the typical range encountered, even
when salinity is changed rapidly. Furthermore, this corroborates my hypothesis that C.
variegatus tolerates extremes in salinity by lowering metabolism and decreasing energy
expenditures. Fish appear to wait for conditions to improve, and respond to these more
favorable conditions by returning metabolism to normal levels.
A decrease in energetic expenditures as just described is a potentially adaptive
response for fishes living in variable salinity environments like those of Florida coastal salt
marshes. While few data exist on responses of other salt marsh residents to wide ranges in
salinity, I suspect this may be a general pattern. Cyprinodon variegatus may be unusual
because of its broad tolerances, but is a useful experimental animal because of this as well.
Information gleaned from studies with C. variegatus may indicate areas of examination for
other important salt marsh teleosts which may have more limited salinity tolerance, but
which may follow the same general metabolic patterns seen in C. variegatus.


CHAPTER 4
INFLUENCE OF A FLUCTUATING SALINITY REGIME ON OSMOREGULATION
IN CYPRINODON VARIEGATUS
Introduction
Few areas in the field of fish physiology have received as much attention as the
study of osmoregulation. The basic patterns of osmoregulation are well understood and are
reviewed extensively by Evans (1984), Kamaky (1986), Ventrella et al. (1992), Evans
(1993), McCormick (1994), and Wood and Marshall (1994). The mechanisms of
osmoregulation in euryhaline fish are not unusual. Euryhaline fishes are unique in having
the ability to osmoregulate efficiently in waters of highly variable salinity. They can
osmoregulate in water both more and less concentrated than their own body fluids, and they
are able to alter their pattern of osmoregulation rapidly if they live in environments where
sudden fluctuations in salinity may occur.
Despite the voluminous research done in this area, few studies have examined
patterns of osmoregulation in fishes that may encounter salinities outside the range from
freshwater (0 ppt) to seawater (35 ppt). Such species tend to be small and of little or no
economic importance, with a notable exception being the milkfish, Chanos chanos (Ferraris
et al., 1988; Swanson, 1991). However, to understand osmoregulation patterns in fishes,
studies need to encompass the range of salinities encountered by species in nature.
One topic that has received little attention concerns the effect of fluctuating salinities
on osmoregulation in tclcosts. Salinity is a limiting factor in certain estuarine and salt marsh
habitats, and may widely vary daily and seasonally. Despite this, most investigations of
osmoregulation in fishes involve studies at constant salinities. While such studies provide
55


56
important information on overall osmoregulatory patterns, they provide less information
about more ecologically relevant responses.
The aim of the present investigation was twofold. First, 1 examined the ability of
individuals of the euryhalinc teleost, Cyprinodon variegatus, to regulate plasma osmolality
under the influence of a cycling salinity regime. Second I examined a hypothesis proposed
by Goolish and Burton (1988) in a study involving the intertidal copepod Tigriopus
californicus. Goolish and Burton (1988) suggested that species exposed to fluctuating
salinities would be able to respond more rapidly and completely to salinity stress. In other
words, could past exposure to changing salinity result not in improved osmoregulation at
any single salinity, but rather to improved performance immediately following another
salinity fluctuation? These hypotheses were examined by determining plasma osmolality
and hematocrit of individual C. variegatus subjected to fluctuations in salinity over a wide
range of ambient salinities.
Methods
Collections of fish used in this study were obtained from tidal creeks in the salt
marsh near Cedar Key, Florida. Specimens were transported back to the laboratory in 128 L
coolers supplied with aeration and filled with water obtained from the collection site. Fish
were obtained in two collections made during September 1994. The salinity of the collection
site was approximately 25 ppt for both collections. This study was conducted at the
Southeastern Biological Science Center (SBSC), National Biological Serv ice, Gainesville,
Florida. Fish were held overnight in the coolers used for transportation (with aeration)
before transfer to a 1.2 m diameter fiberglass holding tank containing water at 30 ppt. Fish
were treated prophylactically for 7-14 days in a 5 mg L'l solution of Acriflavinc.
Following treatment, all fish were transferred to experimental aquaria (30 ppt salinity = 860
mOsm kg'l) located within a constant environment room maintained at 20 1 C and on a


57
12:12 h light:dark cycle. Both holding and experimental aquaria were equipped with sponge
filters providing continuous aeration, and fish were fed flake food once each day.
Experimental aquaria were used to subject fish to a cycling salinity regime. Seven
salinity trials were performed (Table 4-1). All fish were maintained in experimental aquaria
located on bank 1 for eight days prior to initiating the first salinity change. Salinity changes
were effected by carefully netting fish from the experimental aquaria located on bank 1, and
immediately transferring individuals into the appropriate experimental aquaria located on
bank 2. Fish remained in experimental aquaria located on bank 2 for 24 h before transfer
back to tanks located on bank 1. Fish then remained in bank 1 experimental aquaria for 24
h before again being transferred to aquaria on bank 2. Thus, one complete cycle began with
24 h in bank 2 aquaria, and ended following 24 h in bank 1 aquaria The procedure was
repeated so that a total of 10 cycles was performed (20 days). A subsample of fish was
removed for testing from each experimental aquarium just prior to the first change in
salinity (cycle 0), and at the end of cycles 1 (day 2), 5 (day 10), and 10 (day 20). All
subsamples were thus taken following a 24 h period in bank 1 aquaria (30 ppt).
Following completion of the 10th cycle, fish remaining in all decreasing salinity
groups (groups Di, Eb, and D3) were transferred to aquaria at 2 ppt and fish remaining in
all increasing salinity groups (groups Ij, F, and I3) were transferred to aquaria at 60 ppt.
Fish in the aquaria maintained at 30 ppt were split into three groups at that time: one third of
the group was transferred to 2 ppt (group Cd), one third transferred to 30 ppt (group C),
and the final third transferred to 60 ppt (group Q). Fish remained in these salinities for 24
h and were then removed for testing (day 21). Salinities were checked daily with a Leica
temperature compensated refractometer and adjusted as necessary.
Hematocrit (Hct) and plasma osmolality were determined at each sampling interval.
Fish were first carefully netted from their experimental aquaria and blotted dry. Blood was
taken by sternal cardiac puncture using heparinized microhcmatocrit tubes drawn to a fine
point, and fish were weighed and standard length determined. The tubes were then


58
Table 4-1. Salinity trials used in cyclical salinity study. The group maintained at 30 ppt was
split into three groups following cycle 10 (day 20); groups Cq, C, and Q (see text for
details).
Group
Direction of Salinity Change
Salinity in Bank 1
(PPO
Salinity in Bank 2
(PPO
Di
Decreasing Salinity (n=25)
30
2
d2
Decreasing Salinity (n=25)
30
10
d3
Decreasing Salinity (n=25)
30
20
C, Cd, and Q
No Change (n=35)
30
30
It
Increasing Salinity (n=25)
30
40
l2
Increasing Salinity (n=25)
30
50
I3
Increasing Salinity (n=25)
30
60
centrifuged in a micro-hematocrit centrifuge for 10 minutes to separate plasma from cells.
Hematocrit was read using a micro-capillary reader before plasma was isolated from formed
elements by scoring the tube with a file and retaining only the portion containing plasma.
Plasma osmolality (mOsm kg1) was determined on 5 pi samples using a Wescor 5500
vapor pressure osmometer. Fish were used without regard to sex, and all blood samples
were taken between 0600 h and 1000 h. Statistical procedures followed Winer et al. (1991)
and Sokal and Rohlf (1995). All statistical analyses were one way tests using the Tukey-
Kramer post hoc comparison (p = 0.05)
Results
All fish entered into the experimental procedure survived the entire duration of the
experiment. Fish in all groups ate normally throughout the course of the experiment, and no
discernible changes in behavior were observed during the experimental procedure for any


59
group. Fish appeared to have no difficulty in tolerating the imposed salinity fluctuations,
even in groups Di and I3, which were subjected to daily changes in salinity of 28 ppt and
30 ppt, respectively. The influence of body mass on both Hct and plasma osmolality was
evaluated using an analysis of covariance, and for no analysis was body mass a significant
covariate. Sizes of fish used in this study ranged from 0.75 g to 5.887 g (2.8 mm to 5.8 mm
standard length, mean 4.1 0.038 mm), with a mean mass of 2.245 0.066 g. Subsamples
of groups were not significantly different in mass.
Hematocrit and plasma osmolality values were compared both between salinity trials
for each time period sampled (i.e., all groups were compared to one another on days 0, 2,
10, 20, and 21), and within a single trial over the time course of the experiment. Groups
experiencing increases in salinity (groups ty, F, I3, and Q) were compared separately from
groups experiencing decreases in salinity (groups Di, Cty, D3, and Cd), with all compared
to the group maintained at a constant salinity (group C). Data from days 0 through 20 arc
discussed separately from day 21 data as follows.
Day 0 through 20
Hematocrit values showed no obvious trends either between salinity trials or within
any single trial over the time course of the experiment (Table 4-2). There were no
statistically significant differences (p > 0.05) within any group (between days 0 and 20), or
among groups on days 0, 2, 10, or 20. There was a slight trend towards increased variance
in Hct values over the course of the experiment in all groups, peaking at cycle 5, but
differences were non-significant in all cases.
Plasma osmolality data, unlike the Hct data, exhibited consistent trends. No
significant differences existed among groups on days 0, 2, 10, or 20. However, comparisons
within groups over time revealed a different pattern. All groups, regardless of salinity
regime, showed a trend towards slightly elevated plasma osmolality between days 0 and 20
(Table 4-2; Figures 4-1 and 4-2). For groups C and Ij, this was statistically significant, with


Table 4-2. Results of salinity fluctuations experiment. Values in the top row of each cell represent hematocrit measurements (%
erythrocytes), values in bottom row of each cell represent plasma osmolality measurements (mOsm kg*b. Sample sizes are n=5 for each
cell. All values are expressed as means se. See text for explanation of group abbreviations.
GROUP
DAY
D,
d2
d3
Cd
C
Cl
h
b
b
0
24.6 0.68
26.5 0.96
26.4 1.12

25.2 0.92

25.3 1.32
23.8 0.49
25.0 0.63
358.5 332
364.3 7.78
367.3 2.77
364.8 2.33
359.75 8.6
372.7 3.75
375.6 12.7
2
22.8 139
24.8 0.97
23.6 1.03

22.0 0.95

26.0 1.38
27.0 1.64
27.0 1.23
363.3 6.92
379.2 7.97
370.7 10.9
364.5 6.60
380.7 11.2
378.1 7.85
382.4 6.60
10
26.8 1.50
22.8 1.93
23.8 1.28

26.8 1.39

23.0 2.12
28.2 3.25
22.8 2.87
386.1 736
382.6 7.72
379.5 12.0
375.2 3.16
384.6 10.2
389.7 5.48
397.7 9.48
20
24.8 1.02
23.8 1.66
26.0 2.59

24.6 1.40

27.8 2.63
26.4 1.69
24.6 1.69
395.5 7.21
371.6 8.21
403.9 8.20
390.2 2.78
389.8 8.48
398.5 6.69
380.0 7.75
21
26.2 0.49
25.0 1.82
28.8 1.83
25.0 0.84
24.4 1.97
29.2 0.37
27.4 1.12
24.6 1.21
24.2 0.86
367.2 6.81
362.3 9.19
373.1 7.95
336.4 6.48
395.2 4.79
556.6 5.88
436.3 9.3
413.8 9.29
420.8 7.51


Figure 4-1. Mean plasma osmolality values measured for groups experiencing decreases in salinity during the course of the experiment.
Group designations are as follows: Dj, salinity fluctuated between 30 ppt and 2 ppt; D2; salinity fluctuated between 30 ppt and 10 ppt;
D3, salinity fluctuated between 30 ppt and 20 ppt; Co; salinity constant at 30 ppt for days 0-20, decreased to 2 ppt following subsample
on day 20; C, salinity constant at 30 ppt. Sample sizes are n=5 for each group. See text for details of experimental procedure.


c9


Figure 4-2. Mean plasma osmolality values measured for groups experiencing increases in salinity during the course of the experiment.
Group designations are as follows: I j, salinity fluctuated between 30 ppt and 40 ppt; I2; salinity fluctuated between 30 ppt and 50 ppt; I3,
salinity fluctuated between 30 ppt and 60 ppt; Q; salinity constant at 30 ppt for days 0-20, increased to 60 ppt following subsample on
day 20; C, salinity constant at 30 ppt. Sample sizes are n=5 for each group. See text for details of experimental procedure.


Plasma Osmotic Concentration (mOsm leg
to
to _
to
W


65
the values on day 20 significantly higher than values from day 0 and day 1 (px0.05); all
other groups exhibited the same trend, but differences were statistically non-significant.
This appears to be related to the experimental manipulation of fish, and not to the salinity
regime experienced by each group. All groups regulated plasma osmolality effectively. No
differences in regulatory ability could be seen among any of the groups tested, regardless of
the magnitude or direction of the salinity fluctuation.
Day 21
The above addresses the question of how effectively C. variegatus regulates plasma
osmolality in the face of salinity fluctuations. Transfer of individuals to either 2 or 60 ppt
following 20 days of exposure to a variety of salinity fluctuations addresses the second
question posed earlier Does past exposure to large salinity fluctuations result in improved
osmoregulatory performance, compared to animals experiencing little or no salinity
fluctuation, immediately following a fluctuation in salinity? All measurements here were
taken 24 h (on day 21) following the final salinity fluctuation.
Hematocrit results showed no consistent trends on day 21 samples. Only one
significant difference was noted, with the Hct value for group Q being significantly elevated
compared to the value for group C (p<0.05). All other comparisons showed no significant
differences in Hct.
There were significant differences in plasma osmolality for day 21 measurements.
Here groups which had experienced greater fluctuations in salinity over the course of the
experiment showed a much better ability to osmoregulate compared to the control group or
groups which had undergone small salinity fluctuations. Increasing salinity seemed to have
a greater impact on plasma osmolality than did decreasing salinity (Figures 4-1 and 4-2).
Plasma osmolality in groups Dj, [>, and D3 all exhibited declines on day 21
relative to day 20, but these differences were not statistically significant. No differences in
regulatory ability among these three groups, or with the control group (group C), could be


66
discerned. However, group Cq, which had been maintained at 30 ppt for the first 20 days of
the experiment, did show a significant decrease in plasma osmolality on day 21 relative to
values measured on days 0 through 20. Furthermore, this group had lower osmolality
values on day 21 than all other groups, although this difference was only statistically
significant when compared to groups C and D3.
More striking differences existed for groups 11, h, I3, and Q. In all cases plasma
osmolality was elevated on day 21 relative to measurements taken from the same group on
day 20 or earlier. Group I3, which had experienced daily fluctuations in salinity of 30 ppt,
showed the smallest increase in osmolality values, being significantly higher than
measurements from day 0 only. Group U showed a similar result, with the plasma
osmolality value from day 21 significantly higher than values from day 0 and day 2 only.
Groups Ii and C| showed the largest changes in plasma osmolality on day 21. Both groups
had osmolality values that were significantly higher than all previous measurements taken
from their respective groups. The value for group Ii was elevated compared to groups I2,13,
and C, although this increase was statistically significantly only when compared to group C.
Group C| was significantly elevated when compared to all other groups (including group
It).
Discussion
The ability to adjust rapidly to altered salinities would be an obvious advantage to
salt marsh organisms. Physiological responses of euryhaline fishes exposed to rapid
changes in salinity can be grouped into two phases (Holmes and Donaldson, 1969): an
adaptive period and a regulatory period. During the adaptive period, plasma osmolality
varies, gradually returning to values approaching original levels. In the regulatory period,
plasma osmolality is more finely controlled as the fish adjusts to the altered salinity and
reaches ionic homeostasis. Fishes which reach the regulatory period quickly (i.e., have short
adaptive periods) should be best able to tolerate alterations in ambient salinity. Although the


67
length of the adaptive period was not measured in the present study, results nevertheless
suggest that it is relatively short for C. variegatus.
Nordlie (1985) showed that C. variegatus was an excellent osmotic regulator over a
salinity range from 0.3 ppt to 70 ppt. Plasma osmolality values changed only slightly over
this range of salinities in his study, varying by only 40 mOsm kg'l. However, fish in his
study were fully acclimated to each experimental salinity prior to testing. The present study
indicates that C. variegatus is an excellent regulator of plasma osmolality even when fishes
are exposed to large daily fluctuations in salinity. Although fish in all groups in the present
study regulated at slightly higher levels than seen by Nordlie (1985), plasma osmolality
values varied similarly, with differences of less than 40 mOsm kg' 1 for all groups on days
0 through 20. The transfer process itself elicited much of this variation, with slight increases
in plasma osmolality seen over the time course of the experiment. A similar trend was seen
in a study by Woo and Wu (1982) on the red grouper, Epinephelus akaara, and the black
sea bream, My lio macrocephalus.
The influence of a single alteration in salinity on osmoregulatory ability has been
studied for a number of fishes, with changes in plasma osmolality observed in the present
study of similar magnitude to those seen in other curyhaline fishes (Wakeman and
Wohlschlag, 1983; Engel ct al., 1987; Ferraris et al., 1988; Mancera et al., 1993; Yoshikawa
et al., 1993; Altimiras et al., 1994; Tort et al., 1994). Few studies have examined how
fluctuating salinities influence osmoregulation of fishes. The best comparison of my results
are with a study on Blennius pholis by Davenport and Vahl (1979). In their study B. pholis
were exposed to alternating periods of freshwater and seawater, with each period lasting 6 h.
Similar to the results from the present study, plasma osmolality did not vary significantly
over the course of the experiment.
No consistent differences in Hct were observed over the course of the experiment.
Nordlie ct al., (1995) found that C. variegatus efficiently regulates water content over a
wide range of salinities, with a difference of only 4% observ ed between 0 ppt and 100 ppt.


68
These results suggest that there are no large movements of water between the blood and
tissues as a result of fluctuations in salinity over the range tested. The lack of response in
hematocrit in this study may also indicate that within the range of salinities studied C.
variegatus is subject to normal, or tolerance physiological processes, as an increase in
hematocrit values would be expected if C. variegatus were exposed to conditions leading to
resistance processes (Swift, 1982).
It has been hypothesized that fishes occurring in a habitat where salinity often
fluctuates may be able to respond more quickly and completely to alterations in salinity, and
that the limits of tolerance are farther apart if salinity fluctuates periodically (Gunter, 1967;
Spaargaren, 1974; Johnston and Cheverie, 1985). My results provide evidence that prior
exposure to fluctuations in salinity does impart an osmoregulatory advantage. Fishes
previously exposed to large fluctuations in salinity regulated plasma osmolality better than
fishes that had previously experienced no change or small changes in salinity. Increasing
salinity had the greatest impact on regulation of plasma osmolality. Group C|, which had
experienced no prior change in salinity, and group I j, exposed to the smallest fluctuations in
salinity prior to being transferred to 60 ppt showed large increases in plasma osmolality
compared to the control group and groups which had previously experienced large
fluctuations in salinity. Decreasing salinity groups exhibited the same pattern, but
differences in osmolality were less pronounced. Only group Cp, which had not been
previously exposed to fluctuations in salinity, showed a significant decline in plasma
osmolality after transfer to 2 ppt. Salinities between 2 ppt and 35-40 ppt are typically
encountered by this population of C. variegatus in its native habitat, with salinities as high
as 60 ppt rarely encountered. Thus, it is not surprising that transfer to 60 ppt elicited greater
changes in regulation of plasma osmolality.
Variations in salinity imposed on fish in this study caused no dramatic effects. No
adverse behavior or mortalities were noted throughout the course of the experiment. Thus,
despite the differences seen in regulation of plasma osmolality between some of the


69
experimental groups, these differences were not large enough in magnitude to cause
observable distress in the experimental animals. These results indicate that C. variegatus is
well adjusted for life in variable salinity conditions. It would be interesting to compare
results from the present experiments with studies examining the influence of fluctuating
salinity on osmoregulation in fishes that can tolerate changing salinity, but that experience
infrequent salinity variations in their native habitat.


CHAPTER 5
INFLUENCE OF ENVIRONMENTAL SALINITY ON BLOOD OXYGEN LEVELS OF
CYPRINODON VARIEGATUS
Introduction
The sheepshead minnow, Cyprinodon variegatus, is a euryhaline teleost whose
typical habitats are brackish water, coastal salt marshes that experience frequent salinity
fluctuations. Variations in environmental salinity may directly affect the respiratory system
of fishes in at least two ways: by affecting the solubility of oxygen in the water pumped
over the gills, and by affecting the solubility of oxygen dissolved in plasma. Changes in the
ionic composition of bodily fluids may also interact with oxygen to influence tolerance to
variable salinity conditions (Truchot, 1987). Furthermore, fishes in saline water with low
oxygen tension must balance maximizing branchial oxygen diffusion with greater
osmoregulatory demands due to the accompanying increases in ion and water exchange
(Perry and McDonald, 1993). Additionally, the oxygen content of many aquatic habitats is
subject to large natural fluctuations, so oxygen is a potentially limiting factor by itself
(Dejours, 1987; Graham, 1990). This is especially true in shallow waters, where chronic or
periodic hypoxia may be a common phenomenon (Graham, 1990). Salt marsh habitats are
often exposed to hypoxic conditions (Renaud, 1985;Toulmond, 1987).
Cyprinodon variegatus is an extremely competent euryhaline tcleosL Over a range
of salinities from freshwater (0 ppt) to 40 ppt, very small changes in metabolism occur
(Nordlic, et al., 1991; this study, Chapter 2). Both oxygen consumption and critical oxygen
tension (Pc) arc essentially unaffected by changes in salinity over this range. Salinities
above this range cause metabolic adjustments, with increases in Pc and decreases in
metabolism observed (this study, Chapter 2). The mechanisms involved in maintaining
70


71
constant metabolism and Pc at low to moderate salinities, and the alterations of metabolism
and Pc at higher salinities likely involve adjustments in oxygen transport. Not only do high
salinity waters have lowered concentrations of dissolved oxygen, but salinity itself seems to
elicit physiological responses similar to those resulting from lowered levels of oxygen.
Increasing blood oxygen levels with increasing salinity, possibly leveling off at the highest
salinities, could lead to the observed responses in metabolism and Pc in C. variegatus.
Increased blood gas transport is likely the primary mechansim used by most fishes
to increase the amount of oxygen delivered to tissues. Blood oxygen transport in most
teleosts is dependent upon the respiratory pigment hemoglobin. Blood oxygen transport is
normally increased by increasing the concentration of hemoglobin, increasing the number
of erythrocytes in circulation, and/or adjusting the affinity of hemoglobin for oxygen
(Davis, 1975; Wells et al., 1989; Jensen et al., 1993; Perry and McDonald, 1993). I
examined hemoglobin concentration, erythrocyte count, and packed cell volume (hematocrit)
over a wide range of salinities for individuals of C. variegatus to evaluate the influence of
salinity on blood oxygen levels in this extremely curyhaline species.
Methods
Collections of fish, transportation back to the laboratory, and general lab procedures
were performed as described previously. Using the same protocol described earlier, fish
were sequentially acclimated to a series of salinities ranging from 0 ppt to 80 ppt (0 to 2285
mOsm kgl). At the end of the acclimation period, fish were sacrificed to determine
hemoglobin concentration ([Hb]), hematocrit (Hct), and erythrocyte (RBC) count.
Blood Sampling
Fish were first carefully netted from their experimental aquaria and blotted dry.
Blood was taken by sternal cardiac puncture using freshly heparinized microhcmatocrit
tubes drawn to a fine point. Blood from each fish was collected in two microhcmatocrit


72
tubes. Once the first tube was filled with a volume > 20 pi, the blood was immediately
dispensed into a small ceramic crucible. Eppendorf micropipettes were then used to
dispense aliquots of blood from the crucible into test tubes used in the determination of
hemoglobin concentration and erythrocyte count. A second microhematocrit tube was then
filled and used for determination of the hematocrit, and mass and standard length of fish
were then determined. Fish were used without regard to sex, and all blood samples were
taken between 0700 h and 1100 h. Food was withheld from experimental aquaria for 24 h
prior to testing to ensure that all fish were post-absorptive. Statistical procedures followed
Winer et al. (1991) and Sokal and Rohlf (1995). All statistical analyses were one way tests
using the Tukey-Kramer post hoc comparison (p = 0.05).
Hemoglobin Analysis
Hemoglobin concentration was measured spectrophotometrically on 10 pi samples
using the cyanomethemoglobin method (Brown, 1993). As recommended by Innes and
Wells (1985), the cyanomethemoglobin solutions were centrifuged for 10 min at 5000 g
prior to colorimetric determination to remove erythrocyte debris. This method gives reliable
results when used with fish blood (Blaxhall, 1972; Blaxhall and Daisley, 1973; Coburn,
1973; Innes and Wells, 1985).
Erythrocyte Count
Erythrocytes were counted immediately following dilution in test tubes containing
Natt and Herricks solution (Campbell and Murru, 1990). This solution acts as both stain
and dilutent and is routinely used for counting erythrocytes of fish (Campbell and Murru,
1990). A 1:200 dilution was used and cells were counted in an improved Ncubauer
hcmatocytomcter following precautions outlined in Brown (1993).


73
Hematocrit
Hematocrit was measured to determine the packed cell volume of the erythrocytes
contained in the blood. Immediately after filling the second microhematocrit tube one end
was sealed and the tube placed into a micro-hematocrit centrifuge. Tubes were then
centrifuged for 10 min to separate plasma from formed elements. The hematocrit was read
using a micro-capillary reader and expressed as percent erythrocyte.
From the test values obtained, mean corpuscular volume (MCV), mean corpuscular
hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC) were
calculated for each fish as follows (Brown, 1993):
Hcmol MCH_[tWJ)_ MCHC_[HMM^i
RBC/1 RBC/1 Hct
These erythrocyte indices are used to further define the relationship between hemoglobin
content and size of the erythrocyte.
Results
The various measures of blood oxygen arc arranged by salinity group in Table 5-1.
Significant differences over the range of test salinities were found for all parameters except
MCHC. Body mass had no significant influence on any of the measured or calculated
blood oxygen indices.
Salinity exerted the greatest influence on erythrocyte count (Figure 5-1). Values
obtained at 80 ppt and 0 ppt were significantly higher than all other salinities. Erythrocyte
count was next highest at 60 ppt and 70 ppt, being significantly elevated compared to values
in fish acclimated to 2, 10, 20, and 30 ppt. Fish acclimated to salinities from 2 ppt through
50 ppt exhibited no significant differences in erythrocyte count.
Measurements of hemoglobin concentration exhibited a similar pattern, although
fewer significant differences were noted (Figure 5-2). Mean values of fishes acclimated to 0
ppt were highest and significantly different from fish acclimated to salinities from 2 ppt


Table 5-1. Hematocrit (Hct), hemoglobin concentration ([Hb]), erythrocyte count (RBC), mean corpuscular hemoglobin (MCH), mean
corpuscular volume (MCV), and mean corpuscular hemoglobin concentration (MCHC) as a function of salinity for Cyprinodon
variegatus. All values are expressed as means se. See text for further details on blood indices.
Salinity (ppt)
n
Hct (%)
[Hb] (gdl-l)
RBC (x 106 mm*3)
MCH (pg)
MCV (pm3)
MCHC (%)
0
9
29.56 0.90
7.52 0.35
2.79 0.06
26.93 1.08
106.23 3.78
25.52 1.10
2
12
20.33 1.28
5.44 0.22
1.91 0.07
28.50 0.83
105.76 4.90
27.51 1.49
10
10
23.80 1.55
5.23 0.29
1.88 0.12
28.08 1.37
128.51 8.51
22.34 1.13
20
9
19.56 1.19
4.73 0.23
1.71 0.05
27.68 1.15
113.62 4.65
24.69 1.38
30
10
21.10 0.89
5.55 0.39
1.88 0.14
29.80 1.64
114.92 5.04
26.13 1.22
40
12
22.83 1.22
5.85 0.20
2.16 0.06
27.13 0.52
106.58 6.13
26.20 1.35
50
10
22.5 1.21
5.82 0.35
2.06 0.07
28.39 1.54
109.26 4.26
26.39 1.87
60
10
25.2 1.67
6.75 0.40
2.36 0.08
28.45 1.08
106.34 5.05
27.26 1.73
70
10
25.4 1.87
6.09 0.16
2.28 0.08
26.89 0.57
112.09 8.30
24.88 1.46
80
10
22.9 1.27
6.43 0.29
2.81 0.08
23.04 1.06
81.88 4.40
28.89 2.12


Figure 5-1. Mean erythrocyte (RBC) count over a range of salinities in Cyprinodon variegatus (bars indicate se; numerical values
above the points in the figure indicate sample size at each salinity).


Salinity (ppt)
Red Blood Cell Count (x lO^mm
o
-*
ro
CO
o
cn
-*
cn
ro
cn
CO
cn
9 L


Figure 5-2. Mean hemoglobin concentration ([Hb]) over a range of salinities in Cyprinodon vciriegcitus (bars indicate se; numerical
values above the points in the figure indicate sample size at each salinity).


Salinity (ppt)
Hemoglobin Concentration (g dl )
roGo-^tnai-sioocoo
8


79
through 50 ppt. Hemoglobin concentration was also elevated in fish acclimated to 60 ppt, 70
ppt, and 80 ppt, although these values were only significantly higher than the value of fish
acclimated to 20 ppt.
Hematocrit measurements showed less dependence upon salinity (Figure 5-3).
Mean hematocrit was highest in 0 ppt, and was significantly different than mean values of
fish acclimated to all salinities except 60 ppt and 70 ppt. No other significant differences in
hematocrit were noted.
Calculated erythrocyte indices indicated a slightly different pattern. The average
concentration of hemoglobin in the erythrocyte (MCHC) did not vary significantly among
salinity acclimation groups. However, both the average weight of hemoglobin in the
erythrocyte (MCH) and the average volume of the erythrocyte (MCV) were lowest in fishes
acclimated to 80 ppt. MCH was significantly depressed in fish acclimated to 80 ppt when
compared to groups acclimated to 2, 30, 60, and 70 ppt, with MCV values at 80 ppt
significantly lower than values obtained for groups at 10, 20, 30, 50, and 70 ppt.
Discussion
Changes in environmental salinity can exert profound effects on blood oxygen
transport. Increases in salinity confront fishes with the necessity of satisfying oxygen
requirements under conditions of reduced oxygen availability. Fishes may exploit multiple
strategies to optimize blood oxygen transport. The amount of oxygen delivered to the
tissues by the blood per unit time is a product of the cardiac output, the oxygen tension
difference between arterial and venous blood, and the blood oxygen capacitance coefficient
(Jensen, 1991; Jensen ct al., 1993). The capacitance coefficient reflects the hemoglobin's
oxygen transporting properties, and can be adjusted in what have been termed 'qualitative'
and 'quantitative' ways (Jensen, 1991). Regulation of hemoglobin-oxygen (Hb-CH) affinity
represents the primary method for qualitatively altering oxygen carrying capacity, with
control of hemoglobin concentration the primary quantitative mechanism (Jensen, 1991).


Figure 5-3. Mean hematocrit (Hct) over a range of salinities in Cyprinodon variegatus (bars indicate se; numerical values above the
points in the figure indicate sample size at each salinity).


35
30
25
20
15
10
oo
i i 1 | i i i j i i i i j i i i i | i i i i | i i i I | i i i i | i i v | r i i i | i r i i
0 10 20 30 40 50 60 70 80 90
Salinity (ppt)


82
Blood oxygen carrying capacity can also be increased quantitatively by release of stored
erythrocytes, by accelerating maturation of immature erythrocytes, and/or by production of
new erythrocytes (Murad et al., 1990); release of erythrocytes from storage organs (e.g.,
spleen) appears to be the most likely scenario (Soivio et al., 1980; Wells et al., 1989). Fish
exposed to water of changing salinity would be expected to experience variability in their
blood oxygen capacitance coefficient and blood oxygen carrying capacity (Jensen et al.,
1993). Quantitative mechanisms for adjusting blood oxygen carrying capacity were
examined in this study.
Few studies have examined the influence of salinity on oxygen carrying capacity of
fishes. Guernsey and Poluhowich (1975) examined the blood oxygen capacity of American
eels (Anguilla rostrata) acclimated to 0 ppt, 24 ppt, and 34 ppt. As in C. variegatus,
hematocrit was highest in eels acclimated to 0 ppt. However, while oxygen capacity of
acclimated eels was higher in 0 ppt than 34 ppt, the highest oxygen capacity was seen in
eels acclimated to 24 ppt. In a similar study with the cichlid Oreochromis niloticus, Sun et
al., (1995), observed a similar effect of salinity on measures of blood oxygen, with
hemoglobin concentration significantly higher in 0 ppt than in higher salinities (5 to 20
ppt).
Other factors may also contribute to variations in oxygen carrying capacity of
fishes. Hall and Gray (1929) were among the first to note that there is a general correlation
between the habits of fishes and the hemoglobin concentration of their blood. More recent
studies have shown that this generalization also applies to ery throcyte count and hematocrit
(e.g., Haws and Goodnight, 1962; Cobum, 1973; Larsson et al., 1976; Putnam and Freel,
1978). In general, highly active fishes and those that regularly encounter hypoxic conditions
have elevated blood oxygen carrying capacity relative to other species. Values determined
for C. variegatus in the present study arc comparable to other fishes with similar activity
levels, and indicate that C. variegatus docs not possess exceptionally high oxygen carrying


83
capacity at any salinity tested (Hattingh, 1972; Cobum, 1973; Larsson et al., 1976; Putnam
and Freel, 1978; Smit and Hattingh, 1979; Pelster et al., 1988b).
Salinity had a significant effect on blood oxygen carrying capacity in C. variegatus.
However, differences were seen only at the very highest and lowest salinities tested. As
expected, oxygen carry ing capacity increased at high salinities. Elevations were seen at 60
ppt through 80 ppt when compared to measures made at salinities between 2 ppt and 50 ppt.
One finding of interest was indicated by MCV measurements. These results showed that at
80 ppt overall size of erythrocytes had decreased, implying the production and/or release of
smaller, possibly immature erythrocytes. One possible explanation for this is that smaller
erythrocytes have been found to be more effective in oxygen exchange than larger
erythrocytes (Cobum, 1973) due to a larger surface area per unit volume, which may allow a
faster rate of gas exchange. Energetic constraints related to viscosity of the blood may also
play a role. A balance must exist between the advantages for oxygen transport from
increased hematocrit with a disadvantage due to increased viscosity (Wells and Weber,
1991). Hematocrit began to increase as salinity reached 60 ppt, but declined back towards
normal values at 80 ppt. It is possible that at extremely high salinities even small increases
in hematocrit lead to significant viscosity problems. Utilizing smaller erythrocytes at
extreme hypersalinities may help offset this problem.
Unexpectedly, oxygen carrying capacity was highest in the group acclimated to 0
ppt, as indicated by measurements of all blood indices. Although freshwater is tolerated by
C. variegatus, previous work has shown that acclimation to freshwater is difficult and may
result in significant mortality unless decreases in salinity are made in small steps (Nordlic
and Walsh, 1989; Nordlic et al., 1991). Survival in freshwater requires many of the same
responses that arc necessary at extremely high salinities, with both freshwater and
hypcrsalinc conditions imposing difficult osmoregulatory problems for C. variegatus. In
both situations, proliferation of mitochondria rich cells on gill cpithclia is needed to
maintain ionic balance (Evans, 1984; Evans, 1993; Wood and Marshall, 1994). However,


84
this extends the surface involved in ionic exchange at the expense of gill epithelia involved
in gas exchange. Recent studies have shown that such proliferation of mitochondria rich
cells does impair respiratory gas transfer (Bindon et ah, 1994a; Bindon et al., 1994b).
Mechanisms to increase oxygen carrying capacity of the blood would be expected under
such conditions.
However, freshwater conditions differ significantly from hypersaline conditions in
several ways. Most importantly, metabolism is reduced at extreme hypersalinities (this
study, Chapter 2). In conjunction with elevated Pc, depressed metabolism at these
hypersalinities greatly reduces energetic expenditures, partially alleviating the need for
increased oxygen carrying capacity. Thus, whereas measures of blood oxygen are elevated
at salinities of 60 ppt and higher, increases were moderated by a reduction in overall
energetic expenditures. Fish acclimated to 0 ppt exhibit insignificant reductions in
metabolism, so possible increases in oxygen needs in freshwater can not be compensated
for in this manner. Furthermore, a number of studies have indicated that Hb-Oy affinity is
decreased in freshwater conditions (e.g., Benditt et al., 1941; Weber et al., 1976; Woo and
Wu, 1982). Thus, oxygen carrying capacity may have to be increased in quantitative ways if
oxygen affinity of the hemoglobin molecule is decreased in dilute media.
Another finding from the present study was that C. variegatus exhibits little change
in quantitative measures of oxygen carrying capacity over the range of salinities between 2
ppt and 50 ppt. Previous studies have shown that salinities within this range have little
effect on the metabolic rate, Pc, and osmoregulatory ability of C. variegatus (this study,
Chapters 2,3, and 4). However, as oxygen needs would be expected to rise as salinity was
increased over this range, oxygen carrying capacity was expected to increase as well.
Several possibilities may explain why blood oxygen carrying capacity did not
increase over this range of salinities. First, increases in salinity may require little
compensation in oxygen carrying capacity until extreme hypcrsalinitics arc reached.
However, it is more likely that the lack of a response may have been due to the fact that


85
large variations in salinity alone seldom occur under natural conditions. A larger response
would be expected if salinity were varied together with oxygen and/or temperature. Another
possibility is that C. variegatus, like many fish species, may utilize multiple hemoglobins.
Weber (1990) noted that the use of multiple hemoglobins can extend the range of
conditions under which oxygen is transported efficiently, thus enlarging the range of
available habitats. If C. variegatus utilizes such a mechanism, large quantitative changes in
oxygen carrying capacity might not be seen.
Finally, C. variegatus may exhibit mostly qualitative changes in response to salinity,
i.e., the primary response to changing salinity may be better reflected in Hb-C>2 affinity.
Changes in Hb-C>2 affinity as a result of environmental changes are extremely common in
fishes (e.g., Johansen and Weber, 1976; Wells et al., 1980; Weber, 1981), although this has
rarely been examined with respect to salinity. However, studies on Anguilla anguilla
(Weber et al., 1976) and Salmo salar (Mxime et al., 1990) did find that increases in
salinity between freshwater and seawater led to increases in Hb-Cb affinity. A high affinity
hemoglobin molecule might also be advantageous under hypcrsaline conditions, although it
may be ineffective during activity (McMahon, 1988). As Hb-Cb affinity was not measured
in this study, direct correlation with C. variegatus is purely speculative at this time.
Nevertheless, changes in Hb-Cb affinity would correlate well with the large decreases in
energetic expenditures exhibited by C. variegatus at high salinities. However, other factors
could also be involved. First, C. variegatus may exhibit low salt sensitivity over the normal
range of salinities encountered to reduce dependence of blood oxygen affinity on
environmental salinity (Weber et al., 1976). Second, the primary mechanism used to alter
Hb-Cb affinity results from swelling of erythrocytes (Jensen, 1991; Wells and Weber,
1991; Jensen et al., 1993). Swelling of erythrocytes is normally accompanied by a rise in
cell pH and decreased red cell hemoglobin, NTP and ATP concentrations, all of which serve
to increase the oxygen affinity (Wells and Weber, 1991). However, as seen by the MCHC
and MCV values determined in this study, ery throcytes did not swell in response to


86
increased salinity in C. variegatus. Thus if Hb-02 affinity is altered with changes in
environmental salinity, it must be changed in some other manner.
This study clearly indicates that salinity does influence the oxygen carrying capacity
of the blood of C. variegatus. Quantitative differences in hemoglobin concentration,
hematocrit, and erythrocyte count were noted in response to changing salinity. As discussed
above, it also seems likely that salinity may influence qualitative changes in blood oxygen
transport in C. variegatus. Further research is needed to better understand the influence of
salinity on blood oxygen levels in euryhaline teleosts.


CHAPTER 6
SUMMARY AND CONCLUSIONS
This study examined costs associated with life of a teleost in a variable salinity
environment, represented here by a salt marsh. Cyprinodon variegatus was used to examine
the influence of salinity on routine metabolic rate (RMR), critical oxygen tension (Pc),
osmoregulation, and blood oxygen carrying capacity. Results are summarized below.
1) Field measurements in the Cedar Key salt marsh indicated that this habitat undergoes
extensive variation in salinity, temperature, and oxygen.
2) RMR was relatively constant over a range of salinities from 0 ppt to 40 ppt. At higher
salinities RMR began to decline, and was significantly depressed under hypcrsaline
conditions.
3) Following sequential acclimation to experimental salinities, Pc was unaffected by
changes in salinity between 0 ppt and 40 ppt, with Pc increasing at higher salinities.
4) Reduction in metabolism and rise in Pc corresponded well with a reduced ability of C.
variegatus to regulate plasma osmolality efficiently. Osmotic permeability of the gills may
be reduced at high salinities to offset osmotic losses or ionic gains to/from the environment,
indirectly reducing the potential for oxygen uptake as well.
5) Variations in RMR and Pc as a function of environmental salinity observed in this study
suggest that C. variegatus responds to high salinities by reducing energy expenditures.
These responses effectively increases the time C. variegatus can tolerate such conditions,
albeit at a cost of a reduction in energetic processes. This strategy fits the concept of scope
for survival, as described by Hochachka (1990).
6) When C. variegatus was exposed to simulated tidal changes in salinity, RMR was
unaffected in salinity trials where both acclimation and final salinities were in the range
87


88
typically encountered by this population in its native habitat. Where the acclimation or final
salinities were extremely high (50 and 60 ppt) or extremely low (0 ppt), RMR was
depressed.
7) Acclimation state was the most important factor determining the metabolic response to
simulated tidal changes in salinity. However, direction of the salinity change also influenced
metabolism in C. variegatus, with increasing salinity dealt with more efficiently than
decreasing salinity.
8) Simulated tidal experiments corroborate the hypothesis that C. variegatus tolerates
extremes in salinity by lowering metabolism, and hence decreasing energy expenditures.
Following adverse conditions metabolism returns to normal levels.
9) Cyprinodon variegatus is an excellent regulator of plasma osmolality even when
exposed to large fluctuations in salinity within the range of salinities typically encountered.
Daily fluctuations in salinity of up to 30 ppt elicited no significant differences in
osmoregulatory ability when compared to control fish.
10) Prior exposure to fluctuations in salinity does impart an osmoregulatory advantage.
Fishes previously exposed to large fluctuations in salinity regulated plasma osmolality
better than fishes that had previously experienced no or small changes in salinity.
Increasing salinity had a greater impact on regulation of plasma osmolality than did
decreases in salinity.
11) Salinity had a significant effect on blood oxygen carrying capacity in C. variegatus,
although differences were only noted at the very highest (60 to 80 ppt) and lowest (0 ppt)
salinities tested. Oxygen carry ing capacity and all blood indices were highest in the group
acclimated to 0 ppt.
12) C. variegatus exhibited little change in oxygen carrying capacity over the range of
salinities between 2 ppt and 50 ppt. Possible reasons for this include: (a) increases in
salinity may require little compensation in oxygen carrying capacity; (b) detectable changes
may only occur when salinity is varied in conjunction with variations in oxygen and/or


Full Text
A £ PCrfC?
RESPONSES MADE BY THE SALT MARSH TELEOST CYPR1NODON
VARIEGATUS (ATHERINOMORPHA: CYPRINODONTIDAE) TO LIFE IN A
VARIABLE SALINITY ENVIRONMENT
By
DENNIS CHARLES HANEY
A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL
OFTHE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT
OF THE REQUIREMENTS FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY
UNIVERSITY OF FLORIDA

ACKNOWLEDGMENTS
The work described in this dissertation was the focus of my activities for many
years. Throughout this time I have been fortunate to meet and interact with an extraordinary
variety of people, many of whom played integral roles in the successful completion of this
dissertation.
I would first like to acknowledge the assistance of the members of my graduate
committee: Frank Nordlie, Steve Walsh, Brian McNab, Harvey Lillywhite, Ken Sulak, and
Tom Crisman. I would also like to thank a founding member of my committee, Michelle
Wheatly, who, unfortunately for me, left the University of Florida before I was able to
complete my dissertation. All of these people were instrumental in the completion of this
work. In particular, I owe an incredible debt of gratitude to the chair of my committee, Frank
Nordlie. His unfailing friendship, help, and guidance over the years was truly an inspiration
to me, and I can safely say I would not have made it through to the end without him. Steve
Walsh also deserves special recognition for his much-needed assistance. Steve has been a
close friend and colleague for many years, and I am proud to be the first student to complete
a dissertation with Steve as a member of his committee!
So many members of the Zoology Department have helped out that its difficult to
know where to begin. Brent and Sylvia Palmer, John Matter, Frank Hensley (my fusiform
twin!), and Lou Somma were all invaluable in helping me to maintain good mental health
over the years. We spent many hours trying to figure out life, the universe, and everything.
We also had lots of fun looking for unsuspecting fish and herps. Lisa Gregory has been a
friend I could always count on - no matter what! John Binello was my trusted field
companion. We spent many days tramping through the salt marsh (and other bodies of
11

water) in search of numerous fishes including the wily pupfish, sailfin molly, Florida
flagfish, and fat sleeper. Thanks also go to Leo Nico, Adele Hensley, Pam Fuller, Paula
Cushing, Patricia Harrison, Lianna Jarecki, Becky Thompson, Vince DeMarco, Kevin
Baldwin, Ellen Burroughs, Chris Kardish, Mark Hostetler, Doug Weaver, Frank Jordan,
John Anderson, Carol Binello, and everyone else from Zoology!
For the past 4+ years I have worked with many people at the Department of the
Interior's Gainesville laboratory (presently the National Biological Service, formerly U.S.
Fish and Wildlife Service). While working full-time for the past few years certainly slowed
down my progress towards finishing my Ph.D., the extra time was worth it (mostly!). I
formed many new friendships and learned lots of things I wouldn't have otherwise. Jim
Williams showed me that freshwater clams were actually kind of neat. Noel Burkhead
helped remind me that fish were still much cooler! Les Parker, Jayne Brim-Box, and I had
great fun diving in zero visibility water. Howard Jelks, Gary Hill, Ann Foster, Tina Yanchis,
Rob Whiteford, and Bill Stranghoener were all inspiring at one time or another. Special
thanks go to my office mates, Leslie Straub and Cindy Timmerman, for putting up with me
for so long. Cindy especially has been a great friend and confidante.
Last, but certainly not least, I want to thank my family for supporting me all these
years. Mom and Dad never doubted me, though it seemed like I would never finish. Their
encouragement really did help! My brother Scott helped to show me you really can finish a
dissertation and be successful afterwards. For this, and everything else, I can never thank
him enough.
iii

TABLE OF CONTENTS
LIST OF TABLES vi
LIST OF FIGURES vü
ABSTRACT x
CHAPTERS
1 INTRODUCTION 1
Experimental Animal 7
Questions 10
Study Site 11
2 INFLUENCE OF ENVIRONMENTAL SALINITY ON ROUTINE
METABOLIC RATE AND CRITICAL OXYGEN TENSION OF
CYPRINODON VARIEGATUS 12
Introduction 12
Methods 14
Results 18
Discussion 31
3 INFLUENCE OF SIMULATED TIDAL CHANGES IN AMBIENT
SALINITY ON ROUTINE METABOLIC RATE IN CYPRINODON
VARIEGATUS 39
Introduction 39
Methods 40
Results 46
Discussion 46
4 INFLUENCE OF A FLUCTUATING SALINITY REGIME ON
OSMOREGULATION IN CYPRINODON VARIEGATUS 55
Introduction 55
Methods 56
Results 58
Discussion 66
5 INFLUENCE OF ENVIRONMENTAL SALINITY ON BLOOD
OXYGEN LEVELS OF CYPRINODON VARIEGATUS 70
Introduction 70
Methods 71
IV

Results 73
Discussion 79
6 SUMMARY AND CONCLUSIONS 87
APPENDICES 91
1 CRITICAL OXYGEN TENSION FIGURES 91
2 FIELD MEASUREMENTS 102
LITERATURE CITED 108
BIOGRAPHICAL SKETCH 123
v

LIST OF TABLES
Table page
1-1. Phylogenetic classification of the cyprinodontiform fishes (modified from
Parenti, 1981) 8
2-1. Relationships of routine metabolism (RMR), critical oxygen tension (Pc), and
slope in the conformation region at a series of ambient salinities. Values are given
as means ± se 19
2-2. Measurements of oxygen concentration (mg L‘l), salinity (ppt), and temperature
(°C) taken at four sites in the Cedar Key area from June 1990 through June 1991
(see text for details on location of sites). Values are given as means, se (in
parentheses), sample size 32
3-1. Acclimation and final salinities used in simulated tidal change study 42
3-2. Mean routine metabolism (mg Cb h‘l) before (acclimation salinity) and
following (final salinity) a simulated tidal change. Values are given as means ± se.
Groups exhibiting a significant change in metabolism are indicated with an
asterisk 47
4-1. Salinity trials used in cyclical salinity study. The group maintained at 30 ppt was
split into three groups following cycle 10 (day 20); groups Cq, C, and Q (see text
for details) 58
4-2. Results of salinity fluctuations experiment. Values in the top row of each cell
represent hematocrit measurements (% erythrocytes), values in bottom row of each
cell represent plasma osmolality measurements (mOsm kg'l). Sample sizes are
n=5 for each cell. All values are expressed as means ± se. See text for explanation
of group abbreviations 60
5-1. Hematocrit (Hct), hemoglobin concentration ([Hb]), erythrocyte count (RBC),
mean corpuscular hemoglobin (MCH), mean corpuscular volume (MCV), and
mean corpuscular hemoglobin concentration (MCHC) as a function of salinity for
Cyprinodon variegatus. All values are expressed as means ± se. See text for
further details on blood indices 74
vi

LIST OF FIGURES
Figure page
2-1. Mean adjusted routine metabolic rates (RMR) over a range of salinities in
Cyprinodon variegatus (metabolic rates were mass-adjusted using an analysis of
covariance; bars indicate ± se; numerical values above the points in the figure
indicate sample sizes at each salinity) 21
2-2. Relationship between mean adjusted routine metabolic rates (RMR) and mean
plasma osmolality over a range of salinities in Cyprinodon variegatus (metabolic
rates were mass-adjusted using an analysis of covariance; bars indicate ± se;
plasma osmolality data from Nordlie, 1985) 23
2-3. Mean critical oxygen tension (Pc) measurements over a range of salinities in
Cyprinodon variegatus (bars indicate ± se; numerical values above the points in
the figure indicate sample sizes at each salinity) 26
2-4. Relationship between mean adjusted routine metabolic rates (RMR) and critical
oxygen tensions (Pc) over a range of salinities in Cyprinodon variegatus
(metabolic rates were mass-adjusted using an analysis of covariance; bars
indicate ± se) 28
2-5. Relationship between mean critical oxygen tensions (Pc) and mean plasma
osmolality over a range of salinities in Cyprinodon variegatus (bars indicate ± se;
plasma osmolality data from Nordlie, 19&5) 30
2-6. Relationship between mean adjusted routine metabolic rates (RMR) and mean
plasma osmolality over a range of salinities in Adinia xenica (metabolic rates were
mass-adjusted using an analysis of covariance; bars indicate ± se; numerical
values above the points in the figure indicate sample sizes at each salinity) 35
3-1. Schematic diagram of respirometry apparatus used for routine metabolism
experiments. See text for detailed description of system 44
3-2. Results of metabolic trials where salinity was increased over the course of the
trial. Bars represent groups listed in Table 3-2 for which final salinity was greater
than initial salinity. The height of each bar signifies the magnitude of the salinity
change for each metabolic trial and the asterisk indicates at which of the salinities
(for each metabolic trial) the routine metabolic rate (RMR) was highest. The x axis
has no scale and serves only to visually separate groups 49
vii

3-3. Results of metabolic trials where salinity was decreased over the course of the
trial. Bars represent groups listed in Table 3-2 for which final salinity was less
than initial salinity. The height of each bar signifies the magnitude of the salinity
change for each metabolic trial and the asterisk indicates at which of the salinities
(for each metabolic trial) the routine metabolic rate (RMR) was highest. The x axis
has no scale and serves only to visually separate groups 51
4-1. Mean plasma osmolality values measured for groups experiencing decreases in
salinity during the course of the experiment. Group designations are as follows:
Di, salinity fluctuated between 30 ppt and 2 ppt; D2; salinity fluctuated between
30 ppt and 10 ppt; D3, salinity fluctuated between 30 ppt and 20 ppt; Cq; salinity
constant at 30 ppt for days 0-20, decreased to 2 ppt following subsample on day
20; C, salinity constant at 30 ppt. Sample sizes are n=5 for each group. See
text for details of experimental procedure 62
4-2. Mean plasma osmolality values measured for groups experiencing increases in
salinity during the course of the experiment. Group designations are as follows:
Ij, salinity fluctuated between 30 ppt and 40 ppt; W, salinity fluctuated between 30
ppt and 50 ppt; I3, salinity fluctuated between 30 ppt and 60 ppt; Cp, salinity
constant at 30 ppt for days 0-20, increased to 60 ppt following subsample on day
20; C, salinity constant at 30 ppt. Sample sizes are n=5 for each group. See text
for details of experimental procedure 64
5-1. Mean erythrocyte (RBC) count over a range of salinities in Cyprinodon
variegatus (bars indicate ± se; numerical values above the points in the figure
indicate sample sizes at each salinity) 76
5-2. Mean hemoglobin concentration ([Hb]) over a range of salinities in Cyprinodon
variegatus (bars indicate ± se; numerical values above the points in the figure
indicate sample sizes at each salinity) 78
5-3. Mean hematocrit (Hct) over a range of salinities in Cyprinodon variegatus (bars
indicate ± se; numerical values above the points in the figure indicate sample sizes
at each salinity) 81
A1-1. Plot indicating the calculation of the critical oxygen tension (Pc) for an
individual Cyprinodon variegatus in water at 0 ppt 93
A1-2. Plot indicating the calculation of the critical oxygen tension (Pc) for an
individual Cyprinodon variegatus in water at 50 ppt 95
A1-3. Plot indicating the calculation of the critical oxygen tension (Pc) for an
individual Cyprinodon variegatus in water at 100 ppt 97
A1-4. Generalized critical oxygen tension (Pc) plots at each salinity used in the Pc
experiments. Plots were produced by using the mean Pc, mean routine metabolic
rate (RMR), and mean slope in the conformation region for each salinity group..
viii
99

A2-1. Oxygen concentration (mg L"l), salinity (ppt), and temperature (°C) at four
sites in the Cedar Key area taken between June 1990 and June 1991. Values are
given as means, bars indicate ± se. a) Measurements taken on the bottom at site 1;
b) measurements taken on the surface at site 1; c) Measurements taken on the
bottom at site 2; d) measurements taken on the surface at site 2; e) Measurements
taken on the bottom at site 3; 0 measurements taken on the surface at site 3; g)
Measurements taken on the surface at site 4
104
IX

Abstract of Dissertation Presented to the Graduate School
of the University of Florida in Partial Fulfillment of the
Requirements for the Degree of Doctor of Philosophy
RESPONSES MADE BY THE SALT MARSH TELEOST CYPRINODON
VARIEGATUS (ATHERINOMORPHA : CYPR1NODONT1DAE) TO LIFE IN A
VARIABLE SALINITY ENVIRONMENT
By
DENNIS CHARLES HANEY
December, 1995
Chairman: Frank Nordlie
Major Department: Zoology
Cyprinodon variegatus, a common coastal resident of the western Atlantic Ocean
and Gulf of Mexico, lives in ambient salinities ranging from freshwater to 142 ppt. Fish
used in this study were obtained from a Gulf of Mexico salt marsh near Cedar Key, Florida.
In a steady-state experiment, routine metabolic rate (RMR) and critical oxygen tension (Pc)
were determined at salinities ranging from 0 tolOO ppt. Salinities between 0 and 40 ppt had
little influence on RMR or Pc. However, at salinities above 40 ppt, RMR declined, and Pc
increased. The reduction in RMR and rise in Pc correlates with a reduced ability of C.
variegatus to osmoregulate effectively at high salinities. The variations in RMR and Pc at
high salinities suggests that C. variegatus responds by reducing energy expenditures,
effectively increasing the time that individuals can tolerate hypersaline conditions. The
metabolic patterns of C. variegatus as influenced by simulated tidal changes in salinity were
then measured, with RMR unaffected by changes in salinity between 2 and 40 ppt.
However, at extremely high or low salinities metabolism was affected by changes in ambient
x

salinity. Individuals of C. variegatus responded to fluxes at salinity extremes by reducing
general activity and energy expenditures—essentially waiting for conditions to return to
normal, where they responded by increasing metabolic activity. Cyprinodon variegatus
efficiently regulated plasma osmolality, even when fishes were exposed to large fluctuations
in salinity. However, prior exposure to salinity fluctuations did impart an osmoregulatory
advantage. Fish previously exposed to large salinity fluctuations regulated plasma
osmolality better than fish that previously experienced no or small changes in salinity.
Increasing salinity had a greater impact on osmoregulation than did decreasing salinity.
Salinity also had a significant effect on blood oxygen carrying capacity in C. variegatus,
although differences were only noted at the very highest (60 - 80 ppt) and lowest (0 ppt)
salinities tested. Oxygen carrying capacity was highest in the group acclimated to 0 ppt.
Erythrocyte count was the most consistent indicator of the influence of salinity on blood
oxygen, with hematocrit the least consistent measure.
xi

CHAPTER 1
INTRODUCTION
Salinity is a crucial physicochemical factor that exerts an important influence on
aquatic life, particularly on estuarine and salt marsh organisms that are exposed to
unpredictable salinity fluctuations diumally and seasonally. Salt marshes are intertidal beds
of rooted vegetation that are alternately flooded and exposed by rising and falling tides.
Vegetation on higher ground develops a complex network of branching channels through
which water, nutrients, and aquatic organisms move during the tidal cycle. Intertidal salt
marshes are inhospitable yet interesting environments due to their unstable water levels and
to the extreme variability of their physical, chemical, and biological processes (Cooper,
1974; Adam, 1990; Allen and Pye, 1992).
Factors that may influence salinity in salt marshes include precipitation, wind,
frequency and extent of tides, shoreline height, and coastal topography (Remane and
Schlieper, 1971; Wheatly, 1988). Due to the harshness of the physical environment, salt
marshes tend to have low species diversity, but often high abundance of selected species
(McCIusky, 1989; Dunson and Travis, 1994).
Whereas variable salinity habitats are also characterized by fluctuations in other
environmental factors, such as temperature, dissolved oxygen, and pH (Vemberg, 1983;
Wheatly, 1988), the distribution and abundance of fishes in these habitats is largely
determined by salinity (McCIusky, 1989; Davenport and Sayer, 1993; Gill and Potter,
1993). Not surprisingly, animals respond to fluctuations in salinity in complex ways.
Salinity affects osmoregulation, ventilation, metabolism, acid-base balance, growth,
reproduction, development, and other biological processes (Wheatly, 1988). Some of the
primary responses of telcosts to changes in salinity are reviewed here.
1

2
Avoidance is the first line of defense to variable salinity conditions. If behavioral
responses do not sufficiently minimize exposure to variable salinities, aquatic organisms
must rely on physiological and biochemical responses to tolerate environmental changes
(Beitinger and McCauley, 1990). This may be by passive tolerance (osmoconforming) or
active osmoregulation (Truchot, 1987; McClusky, 1989). Only fishes that are capable of
osmoregulation can tolerate wide changes in salinity, and these euryhaline teleosts will be
the focus of the rest of this review.
The basic patterns of osmoregulation in fishes have been extensively reviewed in
recent years (Parvatheswararao, 1970; Eddy, 1982; Evans, 1984; Zadunaisky, 1984;
Kamaky, 1986; Foskett, 1987; Pisam and Rambourg, 1991; Ventrella et al., 1992; Evans,
1993; McCormick, 1994; Wood and Marshall, 1994). Euryhaline teleosts regulate their
blood osmolality at about one-third the concentration of seawater (35 ppt), and thus face
severe osmotic problems whether in freshwater (0 ppt) or seawater. Body fluids of a teleost
in freshwater are hyper-osmotic to the external environment, whereas in seawater they are
hypo-osmotic. Thus, euryhaline fish possess mechanisms for osmoregulating in both
hyper-osmotic and hypo-osmotic conditions.
Teleosts in seawater are susceptible to a loss of body water to the external
environment, and balance water loss by actively drinking large amounts of seawater.
However, both water and salts are absorbed together across the gut. Ingested excess salts
are actively excreted, divalent ions mostly in urine and feces, and monovalent ions by the
gills.
Active excretion of salts by the gills takes place via chloride cells (Zadunaisky, 1984;
Kamaky, 1986; Foskett, 1987; Pisam and Rambourg, 1991). Chloride cells in scawatcr-
acclimatcd fish are located at the base of the secondary gill lamellae, and are large,
columnar-shaped cells usually extending from the basal epithelium to the external
environment. They arc characterized by numerous mitochondria (for this reason they arc
often referred to as "mitochondria rich cells") and an extensive tubular reticulum continuous

3
with the basolateral membrane (Pisam and Rambourg, 1991). The transport enzyme
Na+,K+-ATPase is restricted to this tubular system and to the basolateral membrane. This
ion pump creates a large Na+ gradient (low in the chloride cell cytoplasm) which drives a
NaCl co-transporter by which Cl' enters the cell. The Cl" accumulates sufficiently in the cell
such that it is able to exit to the external environment across the apical membrane, with Na+
following passively down its electrochemical gradient between adjacent chloride cells
(Evans, 1993; McCormick, 1994; Wood and Marshall, 1994).
In freshwater, teleosts continuously face an efflux of salts and an influx of water.
Hence, their osmoregulatory response is to actively transport salts from the external
environment via the gills, to avoid drinking water, and to excrete copious amounts of dilute
urine. Our present understanding of the mechanisms involved in the uptake of Na+ and Cl'
from the environment in freshwater teleosts is somewhat unclear and incomplete. It appears
that Cl" is actively taken up, with Na+ uptake being a passive process. Whether this takes
place via a freshwater-type chloride cell, or through interactions with acid-base balance is
unclear, as evidence exists for both (Wood and Marshall, 1994).
What is unique about osmoregulation in euryhaline fish is not that they possess a
structurally distinct osmoregulatory mechanism, but that they have the ability to function
efficiently in variable salinities. They must not only be able to hypo- and hyper-
osmoregulate, but they must be able to alter their pattern of osmoregulation quickly if they
live where salinity rapidly fluctuates.
Osmoregulation is controlled largely by the endocrine system. In teleosts, hormones
known to exert osmoregulatory effects include prolactin, growth hormone, cortisol,
angiotensin II, arginine vasotocin, atrial natriuretic peptide, thyroid hormones, urotensins,
vasoactive intestinal peptide, insulin, calcitonin, catecholamines and sex steroids (Jenkins,
1981; Bern and Madsen, 1992; Lcloup and Lcbcl, 1993; Sakamoto et al., 1993;Takei, 1993;
McCormick, 1994). The hormone most involved with osmoregulation under freshwater
conditions is prolactin, with arginine vasotocin, urotensin II, catecholamines, and atrial

4
natriuretic peptide also playing roles (Hirano et al., 1987; Bern and Madsen, 1992; Takei,
1993). Control of osmoregulation in seawater is largely via the effects of cortisol and
growth hormone, together with thyroid hormones, angiotensin II, vasoactive intestinal
peptide, atrial natriuretic peptide, and urotensin I (Balment etal., 1987; Bern and Madsen,
1992; Takei, 1993). Although the exact mechanisms and interactive effects of many of these
hormones are unclear, it is well established that both rapid and long term control of
osmoregulation under hyper-osmotic and hypo-osmotic conditions is mediated via the
endocrine system.
Osmoregulation is not the only physiological process affected by salinity. Salinity
adaptation is a complex event that involves a number of physiological and behavioral
responses, including energetics. Many studies have shown that salinity influences
metabolism of fishes, but little agreement exists with respect to the magnitude or direction of
these effects (Nordlie, 1978; Febry and Lutz, 1987; Nordlie et al., 1991; Swanson, 1991;
Morgan and Iwama, 1991; Kirschner, 1993). One reason for a lack of agreement among
studies is that many have focused on the energetic costs associated with osmoregulation,
while ignoring other related physiological and behavioral processes. For example, many
studies have used measured differences in metabolism at different salinities to represent
differences in osmoregulatory costs. The rationale is that osmoregulation is an energetically
costly event, with the energetic costs being related to the osmotic gradient between the fish
and its environment (e.g., Madan Mohan Rao, 1968; Muir and Niimi, 1972; Furspan et al.,
1984; Zadunaisky, 1984). However, this is probably an oversimplification, since other
factors, such as activity, food intake, and permeability changes may also be influenced by
salinity, and in turn, affect metabolism (Swanson, 1991). For this reason metabolic
measurements represent the overall costs associated with living in a particular salinity
environment, with comparisons among salinities reflecting these total costs, not simply the
cost of osmoregulation.

5
Several attempts have been made to categorize the patterns of metabolic responses
of teleosts to altered salinities (Kinne, 1967; Remane and Schlieper, 1971; Nordlie, 1978;
Morgan and Iwama, 1991). One commonly observed pattern is that the lower rates of
metabolism in response to salinity are associated with environments for which the species
(and life stage) are presumably best adapted for, and in which they are normally found.
Teleosts that inhabit widely variable salinity environments are uniquely characterized by
having metabolic rates that are constant over a wide range of salinities. This allows
euryhaline teleosts to avoid the large metabolic costs normally associated with physiological
adjustment to changing salinity.
Another physicochemical parameter that may act synergistically with salinity is
dissolved oxygen. The dissolved oxygen content of many bodies of water is subject to large
natural fluctuations. This is especially true in shallow water, such as salt marshes, where
chronic and/or periodic hypoxia may be common (Renaud, 1985; Dejours, 1987;
Toulmond, 1987; Graham, 1990). Variations in oxygen can dictate the distribution of some
species in aquatic ecosystems (Boutilier, 1990). Changing environmental salinity may
directly influence respiratory function by affecting both the oxygen solubility in water
pumped over the gills and the solubility of gases dissolved in plasma. Changes in the ionic
composition of the body fluids could also interact with oxygen to influence tolerance to
variable salinity conditions (Truchot, 1987).
Exposure of fish to reduced oxygen tensions initiates physiological responses
mostly directed at increasing the amount of oxygen available to the tissues (Boutilier et al.,
1988). The transfer of oxygen from the environment to the tissues can be characterized as a
series of processes. Gill ventilation is the first step, followed in turn by branchial diffusion,
blood oxygen transport, and diffusion into the tissues (the last of which, due to a lack of
available information, will not be discussed further) (Perry and McDonald, 1993).
Gill ventilation is a function of the frequency and depth of breathing, and is
normally increased when low oxygen tensions are encountered. While ventilation docs not

6
appear to be directly limiting to the uptake of oxygen, substantial cost is involved with
increased ventilatory pumping, which ultimately means that any additional oxygen acquired
is used to fuel the ventilatory apparatus itself (Boutilier et al., 1988; McMahon, 1988;
Cameron, 1989; Perry and McDonald, 1993).
Two processes are largely available to increase branchial oxygen diffusion:
increases in functional gill surface area and increases in the mean water to blood oxygen
partial pressure gradient. This tradeoff is particularly important in regard to salinity, as fish
in waters of low oxygen tension must balance the advantage of maximizing branchial
oxygen diffusion with a disadvantage in osmoregulation due to the accompanying increases
in ion and water exchange (Perry and McDonald, 1993). Increasing blood gas transport is
likely the primary route used by most fish to increase the amount of oxygen delivered to the
tissues. Oxygen transport by the blood in teleosts depends on the respiratory pigment
hemoglobin. Blood oxygen transport is normally increased by increasing the concentration
of hemoglobin, increasing the number of erythrocytes in circulation, and/or by adjusting the
affinity of hemoglobin for oxygen (Davis, 1975; Wells et al., 1989; Jensen et al., 1993;
Perry and McDonald, 1993).
All of the processes described above can be modified to optimize oxygen transport
under a variety of environmental conditions. One additional strategy that can be utilized in
conjunction with the above is the lowering of metabolism in concert with reductions in
oxygen. This potentially minimizes the impact of the lowered oxygen tension, but also
reduces aerobic metabolism and therefore the amount of energy available for physiological
processes.
Most fish would be described as metabolic oxygen regulators, as they maintain a
constant metabolic rate over a range of oxygen tensions extending downward from
atmospheric levels to some low level that has been defined as the critical oxygen tension
(Pc). Below the Pc, metabolism is dependent upon oxygen tension, and decreases linearly
with decreases in oxygen. The Pc was probably first documented by Hall (1929), but was

7
not considered an important index for fishes until formalized by Fry (1947). However, this
is an extremely important variable relating to habitat selection and overall metabolic patterns
of fishes, and calculations of the Pc have since been made for a number of species under a
variety of environmental conditions.
Experimental Animal
The subject of this study was the sheepshead minnow, Cyprinodon variegatus.
Cyprinodon variegatus is a member of the family Cyprinodontidae, a large and diverse
family containing over 650 species in 80 genera (Parenti, 1981; Parker and Komfield, 1995;
see Table 1-1). Members of this family are found in fresh, brackish, and salt water, and
distributed pantropically as well as throughout North America. Cyprinodon variegatus is
the type species for both the genus Cyprinodon and the family Cyprinodontidae.
The genus Cyprinodon comprises a group of approximately 30 species of small,
oviparous fishes commonly known as pupfishes. They exhibit remarkable tolerance to
harsh environmental conditions (Miller, 1981) occurring throughout North and Central
America, the Caribbean Sea, and Venezuela (Darling, 1976; Turner and Liu, 1977; Parenti,
1981; Duggins et al., 1983; Barns and Wohlgemuth, 1993). The genus is characterized by
limited genetic divergence and few unique alleles, even in morphologically distinct species
(Kodric-Brown, 1989). Much of the research on the genus has focused on species
inhabiting North American deserts, where the largest concentration of species occurs. Most
of these desert species have small, allopatric distributions (Miller, 1981; Duggins et al.,
1983). Members of this genus also occur in the coastal brackish and marine waters of
eastern North America, where C. variegatus is the dominant pupfish species.
Cyprinodon variegatus is the only pupfish species having an extensive geographic
range. It is found along the Atlantic coast from Massachusetts to the Florida Keys, and
throughout the Gulf of Mexico. A disjunct population occurs along the Yucatan peninsula
(Johnson, 1974; Darling, 1976; Duggins et al., 1983). Populations arc also located in the

8
Table 1-1. Phylogenetic classification of the cyprinodontiform fishes (modified from
Parenti, 1981).
Order Cyprinodontiformes
Suborder Aplocheiloidei
Suborder Cyprinodontoidei
Section 1
Family Profundulidae
Section 2
Division 1
Family Fundulidae
Division 2
Superfamily Poecilioidea
Family Anablepidae
Family Poeciliidae
Superfamily Cyprinodontoidca
Family Goodeidae
Family Cyprinodontidae
Subfamily Cyprinodontinae
Tribe Orestiini
Genus Orestias
Genus Kosswigichthys
Genus Aphanius
Tribe Cyprinodontini
Genus Cyprinodon
Genus Megupsilon
Genus Jordanella
Genus Floridichthys
Genus Cualac

9
Bahamas, West Indies, and Cuba (Duggins et al., 1983). Cyprinodon variegatus has also
been introduced into several areas, where they have negatively impacted native pupfish
species (Echelle and Echelle, 1987; Echelle and Connor, 1989; Kodric-Brown, 1989; Wilde
and Echelle, 1992). Genetic variability within C. variegatus is as large as the amount of
genetic divergence within the entire genus Cyprinodon, with most of this genetic divergence
occurring in populations north of Cape Hatteras—very little genetic divergence is displayed
among southern populations (Darling, 1976; Schwartz et al., 1990).
Cyprinodon variegatus is a numerically dominant and ecologically important
species throughout most of its range, especially in salt marsh and estuarine waters (Kilby,
1955; Simpson and Gunter, 1956; Relyea, 1975; Naughton and Saloman, 1978;
Subrahmanyam and Coultas, 1980; Stout, 1985; Nelson, 1992; Ross and Doherty, 1994).
Due to its importance, C. variegatus has been an important research animal in diverse
disciplines. These include investigations on the species behavior (Itzkowitz, 1974; Itzkowitz,
1978; Mettee and Beckham, 1978; Itzkowitz, 1981, Dwyer and Bculig, 1991), ecology (Doll
and Bast, 1969; Martin, 1970; Martin, 1972; Able, 1976; Harrington and Harrington, 1982;
Fyfe, 1985; Shipley, 1991; Avila etal., 1992; Wright et al., 1993), evolution (Elder and
Turner, 1994), life history (Warlcn, 1964; Dc Vlaming et al., 1978; Berry, 1987; Able, 1990;
Echelle and Echelle, 1994), physiology (Martin, 1968; Karnaky et al., 1976;
Subrahmanyam, 1980; Nordlie, 1985; Barton and Barton, 1987; Nordlie, 1987; Peterson
and Gilmore, 1988; Nordlie and Walsh, 1989; Peterson, 1990; Pnce et al., 1990; Nordlie et
al., 1991; Dunson et al., 1993), and reproduction (Raney etal., 1953; Warlen, 1964; Berry,
1987; Kodric-Brown, 1987; Conover and DcMond, 1991). The species has also been used
in voluminous toxicology experiments because of its extreme hardiness (e.g., Schimniel and
Hansen, 1975; Hawkins et al., 1984; Battalora et al., 1985; Linton, 1992).
Cyprinodon variegatus has been called "the toughest fish in North America"
(Gunter, 1967) due to its extreme tolerance of harsh environmental conditions. It is found in
waters ranging from freshwater (Ager, 1971; Johnson, 1974; Kushlan, 1980) to salinities of

10
142 ppt (Simpson and Gunter, 1956), although it typically inhabits brackish water and
coastal salt marshes. Cyprinodon voriegatus is tolerant of temperatures ranging from about
1 °C (Berry, 1987), to 41 °C (Strawn and Dunn, 1967), and oxygen levels approaching
anoxia (Odum and Caldwell, 1955). Thus, it is an exceedingly useful experimental subject
for studying how teleost species respond to harsh environmental conditions.
Although certain organisms can withstand greater changes in environmental
conditions than others, the ability to respond to natural environmental changes is a basic
characteristic of all living systems. Unfortunately, the terminology used to describe these
responses is not uniform. Various researchers have attempted to define the terms adaptation,
acclimation, acclimatization, and accommodation (e.g., Prosser, 1955; Kinne, 1962; Prosser,
1975; Smit, 1980; Fontaine, 1993). I will use the term "adaptation" in its broadest sense,
defining it as a modification of the characteristics of an organism that facilitate an enhanced
ability to survive and reproduce in a particular environment. In this way I recognize that
adaptations involve both genetic and physiological (phenotypic) components, while not
attempting to separate these components from one another. The term acclimation will be
used as defined by Prosser (1975), where compensatory changes are measured following
changes in single environmental variables.
Questions
This study was designed to examine some of the costs to C. variegatus associated
with living in variable salinity environments. Specifically, I asked the following questions:
(1) What are the metabolic costs associated with different ambient salinities? (2) How does
salinity influence the energetic response at low oxygen tensions? (3) What is the
osmoregulatory response to variable salinity environments? (4) How docs salinity influence
blood oxygen levels? These questions were tested by the following: measurement of
metabolism in C. variegatus fully acclimated to a wide range of experimental salinities;
measurement of the critical oxygen tension in C. variegatus fully acclimated to the same

11
range of salinities; measurement of metabolism prior to, and following, simulated tidal
changes in salinity; monitoring of plasma osmolality in C. variegatus exposed to a group of
different cycling salinity regimes; and measurement of hemoglobin concentration,
erythrocyte count, and hematocrit in C. variegatus acclimated to a wide range of ambient
salinities.
Study Site
Fish used in this study were collected from tidal marshes of the Gulf of Mexico
near Cedar Key, Florida. The shore in the Cedar Key area is classified as a zero energy
sector in which wave energy is dampened over the broad, shallow limestone plateau of the
Gulf of Mexico bottom (Stout, 1985). This results in a wide intertidal zone along the coast.
Furthermore, the coastal physiography is extremely diverse due in large part to irregularities
in the shore line of the mainland, to the presence of numerous islands and oyster bars in the
tidal area, and to the maze of intertidal and subtidal creeks and channels (Kilby, 1955). No
significant sediment sources are found in this area, and tides occur on a semi-diurnal basis.
The dominant emergent vegetation in the area is Spartina alterniflora, with the salt marshes
dominated by Juncus roemerianus. Fish communities of the Juncus marsh are dominated
by atheriniforms, with C. variegatus, Fundulus similis, and Poecilia latipinna making up
50-90% of the catch throughout most of the year (Kilby, 1955; Simpson and Gunter, 1956;
Stout, 1985; pers. obs.).

CHAPTER 2
INFLUENCE OF ENVIRONMENTAL SALINITY ON ROUTINE METABOLIC RATE
AND CRITICAL OXYGEN TENSION OF CYPRINODON VARIEGATUS
Introduction
Most fishes are capable of tolerating only a narrow range of salinities. However,
some fishes live in areas that experience frequent variations in salinity. These euryhaline
species possess important physiological and behavioral mechanisms that enable them to
survive in variable salinity environments. One such fish is the sheepshead minnow,
Cyprinodon variegatus. This species ranges along most of the Atlantic coast of the U.S.,
throughout the Gulf of Mexico, and disjunctly along the Yucatan peninsula (Johnson, 1974;
Darling, 1976; Duggins et al., 1983). It typically inhabits brackish water coastal salt
marshes that undergo frequent salinity fluctuations. Cyprinodon variegatus is capable of
tolerating salinities ranging from 0 ppt (Ager, 1971; Johnson, 1974; Kushlan, 1980) to 142
ppt (Simpson and Gunter, 1956).
This study was designed to examine the metabolic response of C. variegatus over a
variety of environmental salinities. Whereas energetic responses to a number of variables
including temperature, body mass, oxygen, and activity level have been well studied, the
influence of salinity on metabolism of fishes has received less attention. Most previous
studies have found that salinity docs affect the energetics of fishes. Unfortunately, the
magnitude and/or direction of these effects are equivocal (Febry and Lutz, 1987; Morgan
and Iwama, 1991; Nordlic et al., 1991; Swanson, 1991). This has led several authors to
categorize the general patterns of metabolic responses to altered salinities (c.g., Nordlic,
1978; Morgan and Iwama, 1991).
12

13
One general pattern of fishes that inhabit variable salinity environments is a stable
metabolic rate over a range of salinities, with the range of salinities most commonly tested
between freshwater (0 ppt) and seawater (35 ppt) (Morgan and hvama, 1991). Physiological
stability enables such fish to have a euryhaline existence that is unfettered by large
metabolic costs associated with adjustment to salinity change. However, there are relatively
few studies of the metabolic response of euryhaline fishes over an even wider range of
salinities that they encounter in their natural habitats.
Fish metabolism is also strongly influenced by the partial pressure of oxygen
(PO2). Metabolism of fishes is independent of PO2, as they maintain a constant metabolic
rate over a range of PO2 extending downward from high atmospheric levels to some lower
level defined as the critical oxygen tension (Pc). Below the Pc (conformation region),
metabolism depends on oxygen tension and decreases linearly with decreases in oxygen
(Fry, 1947). Oxygen consumption declines at the Pc because the gas exchange system can
no longer supply both the extra demands of the respiratory system and the oxygen
demands of the tissues (Hughes, 1964). Since Pc is a useful metabolic parameter,
calculations of the Pc have been made for a number of fishes under a variety of
environmental conditions (e.g., Hall, 1929; Tang, 1933; Ultsch et al., 1978; Ott et al., 1980;
Subrahmanyam, 1980; Donnelly and Torres, 1988; Rantin et al., 1992; Nonnotte et al.,
1993). However, the influence of ambient salinity on the Pc has not been specifically
studied. This is surprising since salinity and oxygen are important abiotic factors that may
act synergistically in affecting metabolism.
This study examined the influence of a wide range of environmental salinities on
routine metabolic rate (RMR) and Pc in C. variegatus. I hypothesized that both RMR and
Pc would be unaffected by alterations in salinity over the range of salinities commonly
encountered in natural habitats of C. variegatus. Salinities outside this range, but within the
range known to be tolerated, were predicted to cause increases in both parameters.

14
Methods
Fish used in this study were obtained by seining canals and ditches in the salt marsh
near Cedar Key, Florida (Gulf of Mexico). Specimens were transported back to the
laboratory in 19 L carboys containing water from the collection site. Upon arrival at the
laboratory, individuals were held overnight in this water with constant aeration. The
following day, fish were transferred into holding tanks (75 to 114 L aquaria) maintained at
the salinity at which fish were captured, and treated prophylactically for 7-14 days in a 5 mg
L'l solution of Acrifiavine®. Following treatment, groups of approximately 10 fish were
placed into experimental (38 L aquaria) tanks containing water at a salinity within 10 ppt of
that in which they were collected. Both holding and experimental tanks were equipped with
undergravel filtration and constant aeration, and were maintained in rooms on a 12:12
lightrdark cycle. Fish were fed Tetramin® flake food once each day. All experimental
aquaria were located in a constant temperature environment room that maintained aquaria at
20 ±1 °C.
Experimental aquaria were used to acclimate fish to salinities ranging from 0 ppt to
100 ppt (0 to 2860 mOsm kg*l). The initial acclimation period was 14 days, after which
fish were either used in a metabolic trial or were transferred to the next higher or lower
salinity in the series. Salinity changes were in steps of 5 ppt, with smaller increments used
to acclimate fish to 0 ppt. This procedure was repeated until determinations had been made
at all experimental salinities. Water used in the freshwater acclimation was obtained from
wells in Alachua and Levy counties, Florida (mean conductivity = 360 pS cm'l).
Experimental salinities greater than freshwater but less than full seawater were prepared by
diluting filtered Atlantic Ocean seawater (obtained from the C. V. Whitney Laboratory of the
University of Florida, Marincland, Florida) with appropriate quantities of deionized water.
Salinities greater than 35 ppt were produced by supplementing seawater with appropriate

15
amounts of synthetic sea salt (Instant Ocean®). Salinities were monitored daily with an
AO® temperature-compensated refractometer, and adjusted as necessary.
Metabolic determinations were carried out in sealed, opaque flasks ranging in
volume from 0.6 to 1.175 L, with the volume selected based on the size of the fish being
tested. Rate of oxygen consumption was used to measure metabolism, with flasks being
used as closed respirometers. Measures of metabolism were considered to be routine
metabolic rates, as animals were sequestered in such a way as to minimize, but not eliminate
activity (Winberg, 1956; Fry, 1957). Experiments were performed with post-absorptive fish
in a resting state, but fish were unconstrained and capable of spontaneous motor activity
(Winberg, 1956; Fry, 1957). For a relatively inactive species such as C. variegatus
(compared to actively swimming salmonids, for example), RMR is perhaps the most
appropriate measure of metabolism, as it more closely reflects normal activity patterns of the
fish than does either active or standard metabolic rate.
Each flask was sealed with a rubber stopper through which two hypodermic needles
(No. 18) were inserted. Each needle was fitted on the inside with catheter tubing, one with a
piece long enough to reach the bottom of the flask, the other half this length. A 25 ml plastic
syringe filled with water at the salinity of the experimental aquarium was inserted into the
needle fitted with the short piece of catheter tubing, while an empty 10 ml syringe was
inserted into the other needle. A 1 ml water sample was drawn into the empty syringe for
each determination of P02- As each water sample was withdrawn, an equal volume of water
was injected from the filled syringe into the flask. This method of sampling, along with
minor movements by the fish, effectively stirred the water in the flask (Nordlie, pers.
comm.). Determinations of PO2 were made with a Radiometer® oxygen electrode
connected to a Radiometer® PHM 71 acid-base analyzer.
Measurements of the rate of reduction in PO2 were made at 0.5 h intervals, and
continued until fish had depleted the oxygen level to approximately 20 mm Hg (generally 5
to 9 h). Following the final PO2 determination, each fish w’as removed from its flask, damp-

16
dried and weighed to the nearest 0.01 g. All metabolic determinations were made between
0700 and 1900 hours, and fish were not re-used in other metabolic trials.
To ensure that fish were post-absorptive at the time of testing, food was withheld
from experimental aquaria for 24 h prior to beginning a metabolic reading. Respirometers
were filled with water at the salinity of the experimental aquaria, and placed in a water bath
maintained at 20 ±1 °C. The entire metabolic apparatus was located in a small, semi-
darkened room in which no other activity took place. In order to allow time for the fish to
adjust to the respirometers, individuals were placed into the flasks (with constant aeration)
12 to 16 h before beginning a trial. At the beginning of the metabolic trial, aerators were
removed and the flask was sealed.
Calculation of oxygen saturation values for the experimental conditions (taking into
account salinity, temperature, relative humidity, and barometric pressure) were made using
the equations of Truesdale et al., 1955 and both RMR (mg 02 h‘l) and Pc (mm Hg) were
calculated for each fish. Data used for calculation of metabolic rates were limited to values
obtained while the POo in the respirometer was greater than 100 mm Hg, in order to ensure
that these calculations were made at oxygen tensions well above the Pc. All data were used
for calculation of the Pc. Determination of the Pc was made using a BASIC program to
calculate the critical point (Yeager and Ultsch, 1989). Following recommendations by
Yeager and Ultsch (1989), data for each fish were first plotted to ensure that the relationship
was a two-step function, following which the midpoint approximation was used to calculate
the Pc.
Oxygen consumption is strongly influenced by body mass, so RMR values were
mass-adjusted using an analysis of covariance (ANCOVA). Log mass-independent RMR
was used as the dependent variable and log mass as the covariate. Least square means
derived from the ANCOVA were used as adjusted RMR values. It was not possible to
perform an ANCOVA for the Pc values, so calculations of Pc were mass-corrected to the
value of the average mass (3.13 g) of all individuals used in this study. The exponent

17
describing the relationship between mass and metabolism for C. variegatus (Nordlie et al.,
1991), MR = kW0-68, was used to correct oxygen consumption rates. Values were corrected
following the relationship MRc = (Wo0-32)(3.13-a32)(MRo), where MRc is the mass-
corrected oxygen consumption, W0 is the observed mass, and MRo is the observed oxygen
consumption at mass WQ (Ultsch et al., 1978; Cech, 1990). Statistical analyses follow
procedures outlined in Winer et al., (1991) and Sokal and Rohlf (1995). All statistical
analyses were one way tests using the Tukey-Kramer post hoc comparison (p = 0.05), and
values are given throughout as means ± standard error of the mean (se).
Field Measurements
Salt marsh habitats are widely considered to experience unpredictable and
fluctuating abiotic conditions. However, actual physicochemical measurements are
infrequently reported. To address this issue, field measurements were made at four sites in
the Cedar Key area over a one year period. Whenever possible, measurements at each site
were taken both at the surface and on the bottom (generally 1-1.5 m deep). Dissolved
oxygen, salinity, and temperature were measured one to three times each month between
0700 h and 1700 h, for a total of 19 dates between June 1990 and June 1991. Sites 1, 2, and
3 were located deep in the salt marsh where C. variegatus was routinely collected. These
sites were located in close proximity to one another (< 10 m apart), and were interconnected.
Unlike many locations in the salt marsh, these sites were never completely isolated from one
another or from connections to the Gulf of Mexico, even during the lowest tides. Site 4 was
located directly on the Gulf of Mexico, in the town of Cedar Key, approximately five km
from sites 1, 2, and 3. Although C. variegatus is present at site 4, collections were not made
at this location. These field measurements were not intended to indicate the complete ranges
of oxygen, salinity, or temperature experienced by organisms living within the salt marsh.
Thus, the actual ranges of physicochemical conditions experienced by C. variegatus are
most likely greater than reported here. However, these values arc a subset of the conditions

18
experienced by salt marsh inhabitants, and give an indication of some of the variability in
the measured physicochemical parameters.
Results
Routine Metabolism
Mean RMR was calculated for each fish and organized by salinity groups. Mean
values for unadjusted and adjusted RMR (from ANCOVA) are given in Table 2-1, and
adjusted RMR are plotted against ambient salinity in Figure 2-1.
In the range of ambient salinities between freshwater and 40 ppt, adjusted RMR
values were highest at 2 ppt and 40 ppt, being slightly lower and roughly equivalent at the
other measured salinities in this range. At salinities greater than 40 ppt, there was a
progressive decline in adjusted RMR. Overall, adjusted RMR ranged from a maximum of
0.97 mg 02 h‘l at 2 ppt, to a low of 0.64 mg 02 h'l at 100 ppt, representing a 66%
decline. This decline corresponds with a decreased ability to regulate plasma osmolality at
elevated salinities (Figure 2-2; plasma osmolality data are from Nordlie, 1985).
A multiple linear regression analysis was used to generate a predictive model for
describing the effects of both salinity and mass on metabolism. In this model, Log body
mass (Log W; in g) and salinity (S; in ppt) were used as independent variables, and Log
mass-independent RMR (Log MR; in mg O2 h'l) as the dependent variable. The equation
that best described this relationship is
Log MR = -0.296 + 0.548 (Log W) - 0.001 (S) (Fo.l 10 = 85.445; P < 0.0001)
This model described 62% of all variability about the mean, and the random distribution of
the residuals suggests an absence of significant relationships that might have biased the
analysis.

Table 2-1. Relationships of routine metabolism (RMR), critical oxygen tension (Pc), and slope in the conformation region at a
series of ambient salinities. Values arc given as means ± se.
Ambient Salinity
(PPO
n
Mean Body
Mass
(g)
Unadjusted RMR
(mg C>2 h'l)
Adjusted RMR
(mg 02 h'1)
Critical Oxygen
Tension
(mm Hg)
Slope in
Conformation
Region
0
11
3.17 ± 0.232
0.81 ±0.038
0.78 ± 0.025
56.98 ± 6.92
0.0071 ± 0.0024
2
7
3.38 ± 0.258
1.04 ±0.017
0.97 ± 0.027
51.49 ± 5.85
0.0079 ± 0.0039
15
8
2.95 ± 0.438
0.75 ± 0.048
0.78 ± 0.029
53.68 ± 4.92
0.0072 ± 0.0021
30
10
3.57 ± 0.402
0.88 ± 0.033
0.83 ± 0.020
52.16 ± 5.10
0.0058 ± 0.0018
40
9
2.19 ±0.337
0.76 ± 0.050
0.96 ± 0.030
52.14 ± 2.70
0.0079 ± 0.0017
50
18
3.69 ± 0.322
0.94 ± 0.031
0.87 ± 0.020
61.81 ± 4.89
0.0065 ± 0.0011
60
8
3.42 ± 0.491
0.83 ± 0.057
0.80 ± 0.030
63.41 ± 6.06
0.0033 ± 0.0008
70
12
2.70 ± 0.389
0.65 ± 0.050
0.73 ± 0.025
66.31 ± 5.57
0.0041 ± 0.0006
80
12
2.98 ± 0.437
0.67 ± 0.047
0.70 ± 0.025
79.53 ± 5.61
0.0033 ± 0.0004
90
8
3.52 ± 0.362
0.73 ± 0.040
0.68 ± 0.029
74.94 ± 8.00
0.0034 ± 0.0004
100
8
3.21 ± 0.460
0.63 ± 0.032
0.64 ± 0.030
73.93 ± 8.78
0.0034 ± 0.0005

Figure 2-1. Mean adjusted routine metabolic rates (RMR) over a range of salinities in Cyprinodon variegatus (metabolic rates were mass-
adjusted using an analysis of covariance; bars indicate ± se; numerical values above the points in the figure indicate sample size at each
salinity).

Salinity (ppt)
Adjusted Routine Metabolic Rate (mg Oxygen hr )
popo I-!-
k> on be
\Z

Figure 2-2. Relationship between mean adjusted routine metabolic rates (RMR) and mean plasma osmolality over a range of salinities in
Cyprinodon variegatus (metabolic rates were mass-adjusted using an analysis of covariance; bars indicate ± se; plasma osmolality data
from Nordlie, 1985).

Adjusted Routine Metabolic Rate
(mg Oxygen/hr)
Plasma Osmolality (mOsm/kg)

24
Critical Oxygen Tension
Measurements of oxygen consumption (mg 02 h'l) for each fish at all time
intervals were mass-adjusted to the mean mass of all individuals used in this study. These
mass-adjusted values were then used to calculate the Pc for each fish. Mean values of Pc
were organized by salinity group and are given in Table 2-1. Plots showing the relationship
between Pc and ambient salinity and between Pc, RMR, and ambient salinity are shown in
Figures 2-3 and 2-4, respectively.
Mean Pc values in the range of ambient salinities from 0 ppt to 40 ppt were not
significantly different from one another (p = 0.95), similar to the pattern exhibited by the
RMR data Mean Pc values increased at salinities greater than 40 ppt, with the highest levels
recorded at salinities 80 ppt and higher. Pc values ranged from a low of 51.49 mm Hg at a
salinity of 2 ppt, to a high of 79.50 mm Hg at 80 ppt, representing a 45% increase. The rise
in mean Pc values corresponds well with a decreased ability to regulate plasma osmolality,
again similar to the RMR pattern (Figure 2-5; plasma osmolality data are from Nordlie,
1985).
Below the Pc, metabolism depends on the oxygen tension and decreases as the PO2
decreases. Mathematically, the slope of the resulting line in this conformation region
depends on three factors: the Pc, the RMR at the Pc, and the lethal P02- A comparison of
the slopes in the conformation region reveals that despite the changes noted above in RMR
and Pc values, the PO2 at which the fish can no longer survive (under experimental
conditions) is essentially equivalent for all salinities tested. This is reflected in the
increasingly shallow slopes seen at salinities greater than 50 ppt (Table 2-1).
Field Measurements
The field measurements of oxygen concentration, salinity, and temperature revealed
very high variability of these physicochemical parameters over the course of the sampling

Figure 2-3. Mean critical oxygen tension (Pc) measurements over a range of salinities in Cyprinodon variegcitus (bars indicate ± se
numerical values above the points in the figure indicate sample size at each salinity).

Salinity (ppt)
Critical Oxygen Tension (mm Hg)
â– u tn o\ oo \o
© o o o o o
9c
100

Figure 2-4. Relationship between mean adjusted routine metabolic rates (RMR) and critical oxygen tensions (Pc) over a range of salinities
in Cyprinodon variegatus (metabolic rates were mass-adjusted using an analysis of covariance; bars indicate ± se).

Adjusted Routine Metabolic Rate
(mg Oxygen/lir)
0 10 20 30 40 50 60 70 80 90 100 110
Salinity (ppt)
Critical Oxygen Tension (mm Hg)

Figure 2-5. Relationship between mean critical oxygen tensions (Pc) and mean plasma osmolality over a range of salinities in Cyprinodon
variegatus (bars indicate ± se; plasma osmolality data from Nordlie, 1985).

Critical Oxygen Tension (mm Hg)
100
700
90 -
Critical Oxygen Tension
Plasma Osmolality
80 -
70 -
60 -
50 -
40 -\ ' 1 1 1 » 1 1 . 1 . | i , • 1 , | . 1 , , r
0 10 20 30 40 50 60 70 80 90 100
Salinity (ppt)
- 650
- 600
- 550
- 500
- 450
- 400
- 350
-- 300
110
Plasma Osmolality (mOsm/kg)

31
period (Table 2-2). For all sites combined, ranges of dissolved oxygen, salinity, and
temperature over the course of the year were 0.6 - 10.8 mg L'l, 1.0 - 29.0 ppt, and 9.0 -
38.0 °C, respectively. Even higher salinities (> 40 ppt) were encountered occasionally in the
salt marsh outside the sampling period. Other areas of the salt marsh probably encounter
even greater extremes of these variables. Cyprinodon variegatus was seen on all dates when
physicochemical measurements were made.
In addition to high overall variability, there were large differences among some sites
in close proximity to one another. For example, mean oxygen concentration measurements
taken during the Fall at sites one, two, and three were 2.94 mg L‘ 1, 3.85 mg L" 1, and 5.1 mg
L'l, respectively. All three physicochemical parameters also strongly varied temporally
among sampling dates. These data provide good evidence that the Cedar Key salt marsh is
an extremely variable habitat with respect to these physicochemical parameters.
Discussion
The family Cyprinodontidae is a diverse group of fishes with many species that
tolerate extreme environmental conditions (Lowe et al., 1967; Lotan and Skadhauge, 1972;
Naiman et al., 1976; Stuenkel and Hillyard, 1981; Chung, 1982). Cyprinodon variegatus is
perhaps the most physiologically tolerant member of the family. It has been called "the
toughest fish in North America" (Gunter, 1967) due to its extreme tolerance of severe
abiotic conditions. The species is found in waters ranging in salinity from freshwater (Ager,
1971) to 142 ppt (Simpson and Gunter, 1956), and can reproduce in waters as high as 100
ppt (Martin, 1972). They are known to tolerate temperatures ranging from about 1 °C
(Berry, 1987), to temperatures greater than 41 °C (Strawn and Dunn, 1967), and to tolerate
near anoxic conditions (Odum and Caldwell, 1955). Thus, this species is an exceedingly
useful experimental subject for examining how tclcosts respond to harsh environmental
conditions.

Table 2-2. Measurements of oxygen concentration (mg L'l), salinity (ppt), and temperature (°C) taken at four sites in the Cedar Key
area from June 1990 through June 1991 (see text for details on location of sites). Values are given as means, se (in parentheses),
sample size.
Oxygen (mg L"l)
Salinity (ppt)
Temperature (°C)
Location
Depth
Spring
Summer
Fall
Winter
Spring
Summer
Fall
Winte
r
Spring
Summer
Fall
Winter
Site 1
2.18
3.15
2.63
4.24
13.9
16.63
12.75
11.3
25.2
30.50
21.75
14.90
Bottom
(0.28)
(0.54)
(0.42)
(0.97)
(1.05)
(1.45)
(2.62)
(1.69)
(0.37)
(1.03)
(2.39)
(1.35)
n = 5
n = 8
n = 4
n = 5
n = 5
n = 8
n = 4
n = 5
n = 5
n = 8
n = 4
n = 5
3.08
3.72
3.25
5.16
11.00
15.92
13.75
10.60
25.60
31.83
21.63
15.00
Surface
(0.38)
(0.60)
(0.51)
(136)
(1.04)
(2.88)
(2.44)
(0.80)
(1.37)
(1.49)
(2.93)
(1.18)
n = 5
n = 6
n = 4
n = 5
n = 5
n = 6
n = 4
n = 5
n = 5
n = 6
n = 4
n = 5
Site 2
1.98
2.37
2.65
6.78
12.30
13.56
12.50
7.00
24.8
29.36
20.00
14.40
Bottom
(0.47)
(0.70)
(0.63)
(1.33)
(2.65)
(2.31)
(2.73)
(0.91)
(0.84)
(0.69)
(2.81)
(1.54)
n = 5
n = 8
n = 4
n = 5
n = 5
n = 8
n = 4
n = 5
n = 5
n = 8
n = 4
n = 5
3.35
4.83
5.05
8.56
6.2
11.08
7.50
4.20
24.70
30.42
20.13
14.60
Surface
(0.36)
(0.56)
(0.89)
(0.66)
(1.47)
(3.29)
(2.60)
(0.72)
(0.97)
(0.84)
(3.18)
(1.63)
n = 5
n = 6
n = 4
n = 5
n = 5
n = 6
n = 4
n = 5
n = 5
n = 6
n = 4
n = 5
Site 3
3.83
3.46
5.08
8.24
6.80
7.20
7.88
4.60
24.90
30.40
21.38
16.60
Bottom
(0.43)
(1.02)
(0.96)
(0.48)
(1.97)
(2.96)
(3.18)
(0.78)
(0.78)
(1.29)
(3.18)
(1.29)
n = 5
n = 5
n = 4
n = 5
n = 5
n = 5
n = 4
n = 5
n = 5
n = 5
n = 4
n = 5
3.65
5.03
5.15
—
4.50
5.33
3.00
—
25.30
31.17
25.50
—
Surface
(0.25)
(0.73)
(0.15)
—
(0.50)
(1.92)
(2.00)
—
(1.33)
(1.59)
(0.5)
—
n = 3
n = 3
n = 2
—
n = 3
n = 3
n = 2
—
n = 3
n = 3
n = 2
—
Site 4
5.10
5.37
4.35
8.06
17.75
27.36
23.88
15.50
27.0
29.71
22.50
15.10
Middle
(0.68)
(0.85)
(0.60)
(0.98)
(3.13)
(0.38)
(0.52)
(2.42)
(0.51)
(0.38)
(2.76)
(1.36)
n = 5
n = 7
n = 4
n = 5
n = 5
n = 7
n = 4
n = 5
n = 5
n = 7
n = 4
n = 5

33
Field measurements indicated that the Cedar Key salt marsh varies tremendously,
both spatially and temporally, in dissolved oxygen, temperature, and salinity, thus exposing
organisms living within the salt marsh to extreme environmental conditions. Cyprinodon
variegatus tolerates these conditions very well. Salinities between 0 ppt and 40 ppt have
little effect on the energetics of C. variegatus as measured by RMR and Pc- This species
fits into the Typ>e I metabolic response pattern (species exhibiting no significant change in
metabolic rate over a wide range of environmental salinities) as categorized by both Nordlie
(1978) and Morgan and Iwama (1991). A similar pattern characterizes the related salt marsh
resident, Adinia xenica (D.C. Haney, unpublished data), in which RMR was more variable,
but relatively constant over salinities from 0 ppt to 50 ppt (Figure 2-6).
Cyprinodon variegatus exhibits a decline in RMR only at salinities exceeding 40
ppt. This result was somewhat unexpected, but is nearly identical to the pattern found by
Nordlie et al., (1991) in a study that also examined metabolism of C. variegatus over a wide
range of ambient salinities. Nordlie et al., (1991) concluded that the depression in
metabolism at high salinities is probably related to permeability changes of the gill
membrane and/or integument. The point at which metabolism is reduced corresponds well
with a diminished ability of C. variegatus to osmoregulate efficiently. If osmotic
permeability of the gills is reduced at high salinities to help offset ionic influx and osmotic
efflux, the potential for oxygen uptake may be reduced as well (Kristensen and Skadhauge,
1974; Skadhauge, 1974; Davenport and Saycr, 1993). Evidence for this hypothesis comes
from several recent studies. In the first of these, Kultz and Onken (1993) found that overall
in vitro permeability of the opercular membrane of the cichlid Oreochromis mossambicus
was reduced in hypcrsaline media, with a simultaneous reduction in passive ion fluxes.
More direct evidence comes from studies by Bindon et al., (1994a) and Bindon et al.,
(1994b), on the rainbow trout, Oncorhynchus mykiss. In these studies the authors
demonstrated that impairment of respiratory gas transfer coincided with chloride cell
proliferation induced by an osmoregulatory challenge.

Figure 2-6. Relationship between mean adjusted routine metabolic rates (RMR) and mean plasma osmolality over a range of salinities in
Adinia xenica (metabolic rates were mass-adjusted using an analysis of covariance; bars indicate ± se; numerical values above the points
in the figure indicate sample size at each salinity).

Adjusted Routine Metabolic Rate
(Mg Oxygen/hr)
600
Salinity (ppt)
Plasma Osmolality (mOsm/Kg)

36
Changes in activity may also account for a depression in metabolism. In a study on
the milkfish, Chanos chonos, Swanson (1991) showed that a depression in metabolism at
an elevated salinity (55 ppt) was highly correlated with a reduction in activity. This may
have been a factor in this study although the method used here for metabolic determinations
minimized activity of fish at all salinities.
The main focus of this study was determination of Pc over a wide salinity range, as
no other study has investigated to any extent the influence of salinity on this parameter.
Critical oxygen tension can be affected by many factors. These include both physical and
biotic parameters such as temperature, salinity, size and activity, reproductive and nutritional
state, and experimental parameters, such as rate of oxygen depletion or manipulation and
disturbance of fish during measurements (McMahon, 1988).
Like other vertebrates, C. variegatus is a good oxygen regulator over a wide range
of oxygen tensions. Critical oxygen tension in C. variegatus was unaffected by changes in
salinity between 0 ppt and 40 ppt, similar to the result for RMR. However, above 40 ppt, Pc
increased with further increases in salinity. This seems reasonable, as fish might be
expected to encounter difficulties obtaining sufficient oxygen at elevated salinities.
Unfortunately, it is difficult to compare my results with those from other groups of
fishes. While the Pc has been measured for a number of freshwater, marine, and intertidal
fishes, only a few authors have considered the effects of salinity in their analyses. Job
(1969) examined oxygen consumption of Tilapia mossambica (= O. mossambicus) at 0.4
ppt, 12.5 ppt, and 30.5 ppt, and found no effect of salinity on the Pc of small (5 g) fish at
either 15 °C, 30 °C, or 40 °C (Pc approximately 50 mm Hg). Salinity did affect Pc on
larger (80 g) individuals at 15 °C, where the Pc doubled from approximately 50 mm Hg to
100 mm Hg in fish acclimated to 0.4 ppt versus 12.5 ppt or 30.5 ppt. However, Pc
calculations made by Job (1969) were extremely rough approximations, and are difficult to
compare with values in this study.

37
Subrahmanyam (1980) examined the influence of oxygen tension on the metabolic
rate of several salt marsh fishes including C. variegatus, with all measurements made at 25
°C and salinities of 17-21 ppt. All species tested in Subrahmanyam (1980) (C. variegatus,
Poecilia latipinna, Lagodon rhomboides, Leiostornus xanthurus, Fundulus grandis, and
F. similis) were oxygen conformers at oxygen tensions of 85 mm Hg and lower, a
somewhat different response than seen in this study. This difference may be due to the
small sample sizes, and relatively few measurements of oxygen consumption made by
Subrahmanyam (1980). In a study on the golden mullet, Liza aurata, Shusmin (1989),
measured the "oxygen threshold" at salinities ranging from freshwater to 50 ppt. He
showed that the oxygen threshold was relatively constant at all salinities between freshwater
and 40 ppt, with a large increase at 50 ppt. Cyprinodon variegatus did not show this type of
response, as the lethal endpoint does not appear to increase at higher salinities in this
species.
Values of the Pc in this study compare fairly well with Pc measurements for other
groups of fishes. Intertidal marine species generally have lower Pc values than C.
variegatus, ranging from 20 to 26 mm Hg in Paraclinus intergripinnis (Congleton, 1980)
to 30 to 40 mm Hg in Gobius cobitus (Bridges, 1988) and Helcogramma medium (Innes
and Wells, 1985; Pelster et al., 1988; Quinn and Schneider, 1991). A number of African
cichlids ( e.g., Oreochromis niloticus, Cichlasoma urophlhalamus, Erelmodus
cyanosticus, Dimidiochromis compressiceps) have either slightly lower, or roughly
equivalent Pc values to the approximate value (55 mm Hg) displayed by C. variegatus in
this study at most salinities (Ross and Ross, 1983; Becker and Fishelson, 1986; Palacios
and Ross, 1986; Verhcycn ct al., 1994). Donnelly and Torres (1988) found Pc values
ranging from 25 to 50 mm Hg for a number of midwatcr fishes from the eastern Gulf of
Mexico, again values near, or slightly lower than those of C. variegatus. Thus, C.
variegatus has Pc values that arc fairly close to those of ecologically and evolutionarily
diverse tclcosts.

38
The variations in RMR and Pc as a function of environmental salinity observed in
this study suggest that C. variegatus responds to high salinities by reducing energy
expenditures. Observations by myself and others indicate that hypersaline conditions are
encountered infrequently, and likely last for short (days) periods of time. Available evidence
suggests that high salinities are metabolically expensive, whether due to an influence on
activity, osmoregulation, or other physiological and/or behavioral processes. Low oxygen
conditions also commonly occur with high salinities. Decreased metabolism in conjunction
with increased Pc reduces energetic expenditures dramatically at elevated salinities. These
responses effectively increase the time C. variegatus can tolerate adverse conditions, albeit
at a cost of a reduction in energetic processes. However, this is an appropriate response to
harsh conditions that appear at variable and unpredictable time intervals, but that are present
for only short periods of time.
The type of metabolic response to salinity in C. variegatus fits the concept of
"scope for survival" as described by Hochachka (1990). This is a pattern of metabolic
response to environmental stressors characterized by a depression in metabolism,
sometimes below maintenance levels. The advantage of such a reduction in energy
expenditure is essentially the slowing of biological time, enabling survival despite the
temporarily imposed physiological stressor.
Cyprinodon variegatus is an extremely successful euryhaline fish that can survive a
large range of salinities, with typically encountered salinities having little effect on normal
metabolism or on the ability of individuals to osmoregulate efficiently. Fish accomodate
extremely high salinities by reducing energetic expenditures. Such a response increases the
amount of time shcepshead minnows can survive hypcrsaline conditions, enabling them to
"wait out" the difficult conditions.

CHAPTER 3
INFLUENCE OF SIMULATED TIDAL CHANGES IN AMBIENT SALINITY ON
ROUTINE METABOLIC RATE IN CYPRINODON VARIEGATUS
Introduction
Salt marshes often undergo large and rapid salinity fluctuations, a condition that
may significantly affect the distribution and abundance of organisms within these habitats.
Changes in salinity may dramatically influence the energetics of individual fish. Information
on the metabolic costs associated with salinity fluctuations may be useful in explaining or
predicting distribution patterns of some coastal fish species.
Energetic patterns of fishes have been examined relative to a number of abiotic
factors, most notably temperature and oxygen (e.g., Wells, 1935; Fry, 1957; Beamish, 1964;
Brett and Groves, 1979; Stuenkel and Hillyard, 1981; Johnston and Battram, 1993).
Comparatively fewer studies have been considered the influence of salinity on metabolic
patterns of fishes (e.g., Kinne, 1966; Madan Mohon Rao, 1974; Nordlie et ah, 1991;
Swanson, 1991). In these studies, various metabolic responses to salinity have been
reported. Euryhaline fish exhibit one of the few consistent patterns, namely that metabolism
remains relatively unaffected over the range of salinities normally encountered (Nordlie,
1978; Morgan and Iwama, 1991).
Most studies that have examined the influence of salinity on fish energetics have
employed measurements from fish maintained at constant salinities. Data from these
experiments are very useful for understanding the overall metabolic patterns exhibited by
the species tested, as fish were usually fully acclimated to each experimental salinity prior to
testing. However, these experiments provide less information about more ecologically
39

40
relevant responses, as fish in their native habitats are often subject to rapid and dramatic
fluctuations in salinity.
Studies of the influence of salinity fluctuations on metabolism of fishes have
usually focused on species that generally experience maximal fluctuations in salinity only
between freshwater (0 ppt) and seawater (35 ppt) (e.g., Davenport and Vahl, 1979; Von
Oertzen, 1984; Moser and Gerry, 1989; Shusmin, 1989; Moser and Miller, 1994).
However, some salt marsh teleosts regularly encounter salinities outside this range. This
study examined the influence of salinity fluxes on routine metabolic rate (RMR) of the salt
marsh teleost, Cyprinodon variegatus, a species that regularly encounters salinities greater
than 35 ppt.
Fish used in this study were fully acclimated to a series of salinities ranging from 0
ppt to 60 ppt, followed by exposure to a simulated tidal change in salinity. The magnitude,
rate, and direction of salinity changes may be important determinants of how salinity
fluctuations affect metabolism. In this study, the direction of the salinity change in
conjunction with the ambient (acclimation) salinity was the primary focus. The magnitude
of the salinity changes were selected to simulate extremes known to occur in the habitat
from which fish were collected, and the rate was chosen to simulate a normal diurnal tidal
cycle. I predicted that C. variegatus would show minimal changes in RMR when the
salinity was changed within the range commonly encountered by the species. Salinity
changes outside this range were hypothesized to lead to depressions in metabolism.
Methods
Collections of fish, transportation back to the laboratory, and general laboratory
procedures were performed as described previously. Fish were first sequentially acclimated
to a series of salinities ranging from 0 ppt to 60 ppt (0 to 1715 mOsm kg'l). However,
unlike the previous experiments, these procedures were designed to measure RMR before

41
and following simulated tidal changes in salinity. Initial salinities (acclimation) and salinities
following the simulated tidal change (final) are shown in Table 3-1.
Metabolic determinations and changes in salinity were carried out in a flow-through
respirometer with an effective volume of 0.875 L (Figure 3-1). The respirometer was
immersed in a thermoregulated reservoir during metabolic trials that served to maintain a
constant temperature within the respirometer. A submersible pump maintained within a
second thermoregulated reservoir (38 L) was connected to the inlet of the respirometer. This
reservoir was vigorously aerated and was used to supply water to the respirometer. The
respirometer outlet emptied back into this main reservoir. A third reservoir (19 L) serv ing as
a salinity source was connected to the main reservoir via a peristaltic pump. Due to
limitations of the available equipment, the system was closed during oxygen partial pressure
(PO2) determinations. Two ports within the respirometer were used to sample water for
determination of P02- A 10 ml plastic syringe filled with water at the experimental salinity
was inserted into the first port (inlet side), while an empty 10 ml syringe was fitted into the
second port (outlet side). As a 1 ml water sample was drawn into the empty syringe, an
equal volume of water was injected from the filled syringe into the respirometer.
Determinations of PO2 were made with a Radiometer® oxygen electrode connected to a
Radiometer® PHM171 acid-base analyzer.
To begin a metabolic trial, the main reservoir and respirometer were filled with water
at the acclimation salinity. Fish were placed into the respirometer 12 h prior to the
beginning of a metabolic determination. This allowed fish sufficient time to adjust to the
experimental setup. The respirometer was then sealed and immersed in the thermoregulation
reservoir, and the submersible pump was turned on at a flow rate of 1 L min'*. The
following morning, the pump was turned off, the system was closed, and PO2
measurements begun. Measurements of the rate of reduction in PO2 were made at 0.5 to
1.0 h intervals, and continued until fish had depleted the oxygen level to approximately 100
mm Hg (generally 4 to 6 h). Following the final PO2 determination, the system was

42
Table 3-1. Acclimation and final salinities used in simulated tidal change study.
Acclimation Salinity
Final Sal ini tv
*
Oppt
0 ppt (control)
20 ppt
2 ppt
2 ppt (control)
20 ppt
30 ppt
10 ppt
10 ppt (control)
30 ppt
20 ppt
Oppt
2 ppt
20 ppt (control)
40 ppt
50 ppt
30 ppt
2 ppt
10 ppt
30 ppt (control)
50 ppt
40 ppt
20 ppt
40 ppt (control)
60 ppt
50 ppt
20 ppt
30 ppt
50 ppt (control)
60 ppt
40 ppt
60 ppt (control)
reopened, and the submersible pump turned on. At this time, either well water (0 ppt) or a
saline solution of variable concentration was placed into the salinity reservoir. The salt
concentration in this reservoir was determined so that its addition to the main reservoir
would change the salinity of the main reservoir and respirometer to the desired final salinity.

Figure 3-1. Schematic diagram of respirometry apparatus used for routine metabolism experiments. See text for detailed description of
system.


45
The volumes of both reservoirs and the rate of pumping by the peristaltic pump were
adjusted so that the salinity would change at a uniform rate over a 6 h period. The peristaltic
pump was plugged into an automatic timer so that salinity changes took place between 1900
h to 0200 h.
On the second morning the system was re-closed and PO2 measurements were
made at the new (final) salinity. PO2 measurements at the final salinity were thus made 6-
12 h following completion of the salinity fluctuation, a period sufficiently long to be past
the stressful transition period (Von Oertzen, 1984). Following the final P02 determination,
the fish was removed from the respirometer, damp-dried and weighed to the nearest 0.01 g.
All metabolic determinations were made between 0700 h and 1500 h, and fish were not re¬
used in other metabolic trials. Fish were thus held within the respirometer for a total of 40
to 44 h (including adjustment period).
Food was withheld from experimental aquaria for 24 h prior to beginning a
metabolic reading to ensure that fish were post-absorptive at the time of testing. Fish were
not fed while in the respirometer. The entire metabolic apparatus was located in a small,
semi-darkened room in which there was no other activity, with determination of metabolic
rates performed as previously described.
Because a metabolic determination for the two salinity treatments was performed on
the same individual, it was not necessary to mass-correct values in order to perform the
statistical analyses. However, to compare data obtained in this study more easily with
published literature, all RMR measurements were mass-adjusted to the value of the average
mass (2.94 g) of all individuals used in this study. Adjusted RMR values were calculated as
previously described. A repeated measures analysis of variance was used to compare
adjusted RMR's between acclimation and final salinities. Statistical analyses follow
procedures outlined in Winer et al. (1991) and Sokal and Rohlf (1995). All analyses of
variance were one way tests using the Tukey-Kramcr post hoc comparison (p = 0.05).

46
Results
Comparison of RMR's at acclimation and final salinities revealed some interesting
patterns (Table 3-2). There were no significant differences (p > 0.05) in RMR between
acclimation and final salinities in any control trials (n = 4 each). Similarly, when both
acclimation and final salinities (2 ppt, 10 ppt, 20 ppt, 30 ppt, and 40 ppt) were in a range that
is typically encountered by C. variegatus, there was a significant change in RMR in only
one trial. Typical salinities in the wild range from 2-5 ppt through 30-35 ppt, with
hypersaline conditions (35-40 ppt) occurring much more frequently than salinities near 0
ppt (pers. obs.). When the acclimation salinity is high (50 ppt and 60 ppt), all groups
exhibited significant elevation of RMR in the lower, more typical, final salinity. The same
general pattern was seen when the final salinity was high, where fish in two of three groups
had depressed RMR at the highest salinities. RMR was depressed when either the
acclimation or final salinity was 0 ppt. The direction of salinity change strongly influenced
the metabolic response. When salinity was increased over the course of the trial (Figure 3-
2), fish were only affected metabolically at the very highest and lowest salinities, where
metabolism was depressed. When salinity was decreased over the course of the trial (Figure
3-3), fish again showed metabolic depression at extreme salinities, but, overall, more groups
exhibited changes in metabolism with changes in salinity than groups in which salinity was
increased.
Discussion
My results demonstrate that C. variegatus maintains a very stable metabolism when
exposed to typical salinity fluctuations seen in the salt marsh habitat at Cedar Key, Florida.
The experimental procedure resulted in no mortalities during or immediately following any
salinity trial. Metabolism was generally unaffected in salinity trials where both acclimation
and final salinities were in the range typically encountered by C. variegatus at Cedar Key. It

47
Table 3-2. Mean routine metabolism (mg Ch h'l) before (acclimation salinity) and
following (final salinity) a simulated tidal change. Values are given as means ± se. Groups
exhibiting a significant change in metabolism are indicated with an asterisk.
Acclimation
Salinity
(PPO
Acclimation
RMR
(mg 02 h'l)
Final
Salinity
(PPQ
Final
RMR
(mg C>2 h*l)
P value
Response
Qppt
0.466 ± 0.07
20 ppt (n = 5)
0.584 ± 0.09
0.04 *
Increase
2 ppt
1.242 ±0.18
20 ppt (n = 5)
1.244 ±0.23
0.99
No Change
1.024 ±0.13
30 ppt (n = 5)
0.978 ± 0.06
0.69
No Change
10 ppt
1.084 ±0.20
30 ppt (n = 5)
1.082 ±0.19
0.97
No Change
20 ppt
1.348 ±0.14
0 ppt (n = 5)
0.906 ± 0.09
0.01 *
Decrease
0.886 ± 0.03
2 ppt (n = 5)
0.478 ± 0.07
0.01 *
Decrease
0.948 ± 0.08
40 ppt (n = 6)
0.785 ± 0.17
0.33
No Change
1.025 ±0.11
50 ppt (n = 6)
0.745 ± 0.07
0.01 *
Decrease
30 ppt
1.028 ± 0.23
2 ppt (n = 5)
0.854 ± 0.10
0.33
No Change
0.628 ± 0.07
10 ppt (n = 5)
0.688 ± 0.11
0.34
No Change
0.728 ± 0.07
50 ppt (n = 6)
0.728 ±0.12
1.00
No Change
40 ppt
0.704 ±0.12
20 ppt (n = 5)
0.662 ± 0.09
0.75
No Change
0.626 ± 0.06
60 ppt (n = 5)
0.476 ± 0.03
0.03 *
Decrease
50 ppt
0.620 ± 0.04
20 ppt (n = 5)
0.904 ±0.11
0.05 *
Increase
0.850 ±0.12
30 ppt (n = 5)
1.076 ±0.14
0.05*
Increase
60 ppt
0.452 ± 0.07
40 ppt (n = 5)
0.578 ± 0.05
0.01 *
Increase
was only when the acclimation or final salinities were outside the range normally
encountered (i.c., 0 ppt, 50 ppt, and 60 ppt) that changes in metabolism were found. In these
cases metabolism was depressed at each extreme salinity relative to the measurements made
at the more typical salinity.

Figure 3-2. Results of metabolic trials where salinity was increased over the course of the
trial. Bars represent groups listed in Table 3-2 for which final salinity was greater than
initial salinity. The height of each bar signifies the magnitude of the salinity change for each
metabolic trial and the asterisk indicates at which of the salinities (for each metabolic trial)
the routine metabolic rate (RMR) was highest. The x axis has no scale and serves only to
visually separate groups.

Salinity (ppt)
49
60
50
40
30
20
10
2
0
RMR's Not Significantly Different (p = 0.05)
RMR's Significantly Different (p = 0.05)
*
*
*
Salinity Increasing

Figure 3-3. Results of metabolic trials where salinity was decreased over the course of the
trial. Bars represent groups listed in Table 3-2 for which final salinity was less than initial
salinity. The height of each bar signifies the magnitude of the salinity change for each
metabolic trial and the asterisk indicates at which of the salinities (for each metabolic trial)
the routine metabolic rate (RMR) was highest. The x axis has no scale and serves only to
visually separate groups.

Salinity (ppt)
51
60
50
40
30
20
10
2
0
RMR's Not Significantly Different (p = 0.05)
RMR's Significantly Different (p = 0.05)
* *
H
*
Salinity Decreasing

52
These results allow for limited comparison because similar studies have examined
the influence of salinity over a much narrower range than examined in this study. The best
comparison is with a study by Wakeman and Wohlschlag (1983) on the red drum,
Sciaenops ocellatus. In their study, test animals were acclimated to a series of salinities
between 1 ppt and 50 ppt, and then abruptly transferred to salinities either 10 ppt higher or
lower than the acclimation salinity. Although Wakeman and Wohlschlag (1983) did not
specifically compare metabolic rates following the transfer with those taken at the
acclimation salinity, there were trends similar to results of this study. Fluctuations in salinity
over a moderate range caused little or no change in metabolism of S. ocellatus on
determinations made 12 to 24 h following the transfer. Only when either the acclimation or
final salinity was 50 ppt were changes in metabolism obvious, with metabolic rates elevated
in 50 ppt compared to 40 ppt. It thus appears that 5. ocellatus responds to abrupt salinity
changes somewhat differently than docs C. variegatus, as metabolism increased upon
transfer to 50 ppt in S. ocellatus, but decreased in C. variegatus. This difference may be
due to the smaller magnitude of the salinity changes performed by Wakeman and
Wohlschlag (1983) (10 ppt vs 20 to 30 ppt in the present study). However, these results
could also be attributable to different sizes and activity patterns of experimental fish
between the two studies.
In a similar study, Shushmin (1989) studied the dynamics of oxygen consumption
in juvenile golden mullet, Liza aurata, following abrupt transfers from 18 ppt to a series of
salinities ranging from 0.4 ppt to 50 ppt. All salinity changes resulted in increased
metabolic rates for one to three days following the transfer, with metabolism generally
returning to near normal levels within three to four days. It is not surprising that salinity
changes arc more stressful in the golden mullet, since the species has lower salinity
tolerance and osmoregulatory ability than C. variegatus.
Barton and Barton (1987) examined metabolism in juvenile (< 1 g) C. variegatus
collected from inland saline lakes of San Salvador Island. They measured metabolic rates of

53
fish at 10 ppt and 35 ppt following abrupt transfer from an acclimation salinity of 35 ppt.
Metabolic rates in fish measured at 10 ppt were significantly higher than those made at the
acclimation salinity. These results are somewhat different from those obtained in the present
study. However, measurements by Barton and Barton (1987) were made only 3 h following
abrupt transfer to the reduced salinity, whereas measurements in the present study were
made 6-12 h following a gradual change in salinity. Fish in this study thus experienced a
less stressful transition to the new salinity. An ontogenetic effect may also account for the
observed differences, as many investigations have demonstrated that adults and juveniles of
the same species may differ greatly in physiological competence (e.g., Kinne, 1966; Martin,
1968; Oikawa et al., 1991).
Other relevant comparisons can be made with data on juvenile croaker,
Micropogonias undulatus, and juvenile spot, Leiostomus xanthurus (Moser and Gerry,
1989; Moser and Miller, 1994). These studies examined effects on metabolism from
fluctuations in salinity between 0 ppt and 35 ppt. Like C. variegatus, a salinity decrease
from higher salinities to 0 ppt generally led to a decreased metabolism in L. xanthurus.
Similarly, when fish were acclimated to 0 ppt and salinity was increased, metabolism was
elevated at the higher salinities. Contrary to findings for C. variegatus, salinity fluctuations
between 15 ppt and 35 ppt elicited changes in metabolism in both L. xanthurus and M.
undulatus. Thus, these species are influenced to a greater extent by salinity fluctuations than
C. variegatus. However, rates of salinity fluctuation in the above studies were faster than
rates used in the present study (16 or 32 ppt h*l vs 3.3 or 5 ppt h‘l), and C. variegatus
likely experiences greater fluctuations in salinity than do spot or croaker.
Acclimation state is the most important measured factor influencing the metabolic
response of C. variegatus to simulated tidal changes in salinity. However, direction of the
salinity change also influenced metabolism in C. variegatus. Like the results obtained by
Moser and Miller (1994) on juvenile L. xanthurus, it appears that C. variegatus adjusts to
increasing salinity more effectively than to decreasing salinity, as evidenced by the

54
responses to salinity changes between 20 ppt and 2 ppt, and between 50 ppt and 30 ppt.
Thus, when salinity is increased over the course of the trial, fish are only affected
metabolically at the very' highest and lowest salinities. However, when fish are acclimated to
a high salinity, and salinity is then decreased to a more typical range, individuals take
advantage by increasing metabolism to more normal levels.
Thus, C. variegatus is well adapted to a varying salinity environment. Its
metabolism is unaffected by changes in salinity over the typical range encountered, even
when salinity is changed rapidly. Furthermore, this corroborates my hypothesis that C.
variegatus tolerates extremes in salinity by lowering metabolism and decreasing energy
expenditures. Fish appear to wait for conditions to improve, and respond to these more
favorable conditions by returning metabolism to normal levels.
A decrease in energetic expenditures as just described is a potentially adaptive
response for fishes living in variable salinity environments like those of Florida coastal salt
marshes. While few data exist on responses of other salt marsh residents to wide ranges in
salinity, I suspect this may be a general pattern. Cyprinodon variegatus may be unusual
because of its broad tolerances, but is a useful experimental animal because of this as well.
Information gleaned from studies with C. variegatus may indicate areas of examination for
other important salt marsh teleosts which may have more limited salinity tolerance, but
which may follow the same general metabolic patterns seen in C. variegatus.

CHAPTER 4
INFLUENCE OF A FLUCTUATING SALINITY REGIME ON OSMOREGULATION
IN CYPRINODON VARIEGATUS
Introduction
Few areas in the field of fish physiology have received as much attention as the
study of osmoregulation. The basic patterns of osmoregulation are well understood and are
reviewed extensively by Evans (1984), Kamaky (1986), Ventrella et al. (1992), Evans
(1993), McCormick (1994), and Wood and Marshall (1994). The mechanisms of
osmoregulation in euryhaline fish are not unusual. Euryhaline fishes are unique in having
the ability to osmoregulate efficiently in waters of highly variable salinity. They can
osmoregulate in water both more and less concentrated than their own body fluids, and they
are able to alter their pattern of osmoregulation rapidly if they live in environments where
sudden fluctuations in salinity may occur.
Despite the voluminous research done in this area, few studies have examined
patterns of osmoregulation in fishes that may encounter salinities outside the range from
freshwater (0 ppt) to seawater (35 ppt). Such species tend to be small and of little or no
economic importance, with a notable exception being the milkfish, Chanos chanos (Ferraris
et al., 1988; Swanson, 1991). However, to understand osmoregulation patterns in fishes,
studies need to encompass the range of salinities encountered by species in nature.
One topic that has received little attention concerns the effect of fluctuating salinities
on osmoregulation in tclcosts. Salinity is a limiting factor in certain estuarine and salt marsh
habitats, and may widely vary daily and seasonally. Despite this, most investigations of
osmoregulation in fishes involve studies at constant salinities. While such studies provide
55

56
important information on overall osmoregulatory patterns, they provide less information
about more ecologically relevant responses.
The aim of the present investigation was twofold. First, 1 examined the ability of
individuals of the euryhalinc teleost, Cyprinodon variegatus, to regulate plasma osmolality
under the influence of a cycling salinity regime. Second I examined a hypothesis proposed
by Goolish and Burton (1988) in a study involving the intertidal copepod Tigriopus
californicus. Goolish and Burton (1988) suggested that species exposed to fluctuating
salinities would be able to respond more rapidly and completely to salinity stress. In other
words, could past exposure to changing salinity result not in improved osmoregulation at
any single salinity, but rather to improved performance immediately following another
salinity fluctuation? These hypotheses were examined by determining plasma osmolality
and hematocrit of individual C. variegatus subjected to fluctuations in salinity over a wide
range of ambient salinities.
Methods
Collections of fish used in this study were obtained from tidal creeks in the salt
marsh near Cedar Key, Florida. Specimens were transported back to the laboratory in 128 L
coolers supplied with aeration and filled with water obtained from the collection site. Fish
were obtained in two collections made during September 1994. The salinity of the collection
site was approximately 25 ppt for both collections. This study was conducted at the
Southeastern Biological Science Center (SBSC), National Biological Serv ice, Gainesville,
Florida. Fish were held overnight in the coolers used for transportation (with aeration)
before transfer to a 1.2 m diameter fiberglass holding tank containing water at 30 ppt. Fish
were treated prophylactically for 7-14 days in a 5 mg L'l solution of Acriflavinc®.
Following treatment, all fish were transferred to experimental aquaria (30 ppt salinity = 860
mOsm kg'l) located within a constant environment room maintained at 20 ± 1 °C and on a

57
12:12 h light:dark cycle. Both holding and experimental aquaria were equipped with sponge
filters providing continuous aeration, and fish were fed flake food once each day.
Experimental aquaria were used to subject fish to a cycling salinity regime. Seven
salinity trials were performed (Table 4-1). All fish were maintained in experimental aquaria
located on bank 1 for eight days prior to initiating the first salinity change. Salinity changes
were effected by carefully netting fish from the experimental aquaria located on bank 1, and
immediately transferring individuals into the appropriate experimental aquaria located on
bank 2. Fish remained in experimental aquaria located on bank 2 for 24 h before transfer
back to tanks located on bank 1. Fish then remained in bank 1 experimental aquaria for 24
h before again being transferred to aquaria on bank 2. Thus, one complete cycle began with
24 h in bank 2 aquaria, and ended following 24 h in bank 1 aquaria The procedure was
repeated so that a total of 10 cycles was performed (20 days). A subsample of fish was
removed for testing from each experimental aquarium just prior to the first change in
salinity (cycle 0), and at the end of cycles 1 (day 2), 5 (day 10), and 10 (day 20). All
subsamples were thus taken following a 24 h period in bank 1 aquaria (30 ppt).
Following completion of the 10th cycle, fish remaining in all decreasing salinity
groups (groups Di, Eb, and D3) were transferred to aquaria at 2 ppt and fish remaining in
all increasing salinity groups (groups Ij, F, and I3) were transferred to aquaria at 60 ppt.
Fish in the aquaria maintained at 30 ppt were split into three groups at that time: one third of
the group was transferred to 2 ppt (group Cd), one third transferred to 30 ppt (group C),
and the final third transferred to 60 ppt (group Q). Fish remained in these salinities for 24
h and were then removed for testing (day 21). Salinities were checked daily with a Leica®
temperature compensated refractometer and adjusted as necessary.
Hematocrit (Hct) and plasma osmolality were determined at each sampling interval.
Fish were first carefully netted from their experimental aquaria and blotted dry. Blood was
taken by sternal cardiac puncture using heparinized microhcmatocrit tubes drawn to a fine
point, and fish were weighed and standard length determined. The tubes were then

58
Table 4-1. Salinity trials used in cyclical salinity study. The group maintained at 30 ppt was
split into three groups following cycle 10 (day 20); groups Cq, C, and Q (see text for
details).
Group
Direction of Salinity Change
Salinity in Bank 1
(PPO
Salinity in Bank 2
(PPO
Di
Decreasing Salinity (n=25)
30
2
d2
Decreasing Salinity (n=25)
30
10
d3
Decreasing Salinity (n=25)
30
20
C, Cd, and C[
No Change (n=35)
30
30
It
Increasing Salinity (n=25)
30
40
¡2
Increasing Salinity (n=25)
30
50
i3
Increasing Salinity (n=25)
30
60
centrifuged in a micro-hematocrit centrifuge for 10 minutes to separate plasma from cells.
Hematocrit was read using a micro-capillary reader before plasma was isolated from formed
elements by scoring the tube with a file and retaining only the portion containing plasma.
Plasma osmolality (mOsm kg‘1) was determined on 5 pi samples using a Wescor® 5500
vapor pressure osmometer. Fish were used without regard to sex, and all blood samples
were taken between 0600 h and 1000 h. Statistical procedures followed Winer et al. (1991)
and Sokal and Rohlf (1995). All statistical analyses were one way tests using the Tukey-
Kramer post hoc comparison (p = 0.05)
Results
All fish entered into the experimental procedure survived the entire duration of the
experiment. Fish in all groups ate normally throughout the course of the experiment, and no
discernible changes in behavior were observed during the experimental procedure for any

59
group. Fish appeared to have no difficulty in tolerating the imposed salinity fluctuations,
even in groups Di and I3, which were subjected to daily changes in salinity of 28 ppt and
30 ppt, respectively. The influence of body mass on both Hct and plasma osmolality was
evaluated using an analysis of covariance, and for no analysis was body mass a significant
covariate. Sizes of fish used in this study ranged from 0.75 g to 5.887 g (2.8 mm to 5.8 mm
standard length, mean 4.1 ± 0.038 mm), with a mean mass of 2.245 ± 0.066 g. Subsamples
of groups were not significantly different in mass.
Hematocrit and plasma osmolality values were compared both between salinity trials
for each time period sampled (i.e., all groups were compared to one another on days 0, 2,
10, 20, and 21), and within a single trial over the time course of the experiment. Groups
experiencing increases in salinity (groups ty, F, I3, and Q) were compared separately from
groups experiencing decreases in salinity (groups Di, Cty, D3, and Cd), with all compared
to the group maintained at a constant salinity (group C). Data from days 0 through 20 arc
discussed separately from day 21 data as follows.
Day 0 through 20
Hematocrit values showed no obvious trends either between salinity trials or within
any single trial over the time course of the experiment (Table 4-2). There were no
statistically significant differences (p > 0.05) within any group (between days 0 and 20), or
among groups on days 0, 2, 10, or 20. There was a slight trend towards increased variance
in Hct values over the course of the experiment in all groups, peaking at cycle 5, but
differences were non-significant in all cases.
Plasma osmolality data, unlike the Hct data, exhibited consistent trends. No
significant differences existed among groups on days 0, 2, 10, or 20. However, comparisons
within groups over time revealed a different pattern. All groups, regardless of salinity
regime, showed a trend towards slightly elevated plasma osmolality between days 0 and 20
(Table 4-2; Figures 4-1 and 4-2). For groups C and ty, this was statistically significant, with

Table 4-2. Results of salinity fluctuations experiment. Values in the top row of each cell represent hematocrit measurements (%
erythrocytes), values in bottom row of each cell represent plasma osmolality measurements (mOsm kg*b. Sample sizes are n=5 for each
cell. All values are expressed as means ± se. See text for explanation of group abbreviations.
GROUP
DAY
D,
d2
d3
Cd
C
Cl
h
h
b
0
24.6 ± 0.68
26.5 ± 0.96
26.4 ± 1.12
—
25.2 ± 0.92
—
25.3 ± 1.32
23.8 ± 0.49
25.0 ± 0.63
358.5 ± 332
364.3 ± 7.78
367.3 ± 2.77
364.8 ± 2.33
359.75 ± 8.6
372.7 ±3.75
375.6 ± 12.7
2
22.8 ± 139
24.8 ± 0.97
23.6 ± 1.03
—
22.0 ± 0.95
—
26.0 ± 1.38
27.0 ± 1.64
27.0 ± 1.23
363.3 ± 6.92
379.2 ± 7.97
370.7 ± 10.9
364.5 ± 6.60
380.7 ± 11.2
378.1 ±7.85
382.4 ± 6.60
10
26.8 ± 1.50
22.8 ± 1.93
23.8 ± 1.28
—
26.8 ± 1.39
—
23.0 ± 2.12
28.2 ± 3.25
22.8 ± 2.87
386.1 ±736
382.6 ± 7.72
379.5 ± 12.0
375.2 ±3.16
384.6 ± 10.2
389.7 ± 5.48
397.7 ± 9.48
20
24.8 ± 1.02
23.8 ± 1.66
26.0 ± 2.59
—
24.6 ± 1.40
—
27.8 ± 2.63
26.4 ± 1.69
24.6 ± 1.69
395.5 ±7.21
371.6 ±8.21
403.9 ± 8.20
390.2 ± 2.78
389.8 ± 8.48
398.5 ± 6.69
380.0 ± 7.75
21
26.2 ± 0.49
25.0 ± 1.82
28.8 ± 1.83
25.0 ± 0.84
24.4 ± 1.97
29.2 ± 0.37
27.4 ± 1.12
24.6 ± 1.21
24.2 ± 0.86
367.2 ± 6.81
362.3 ± 9.19
373.1 ±7.95
336.4 ± 6.48
395.2 ± 4.79
556.6 ± 5.88
436.3 ± 9.3
413.8 ± 9.29
420.8 ±7.51

Figure 4-1. Mean plasma osmolality values measured for groups experiencing decreases in salinity during the course of the experiment.
Group designations are as follows: Dj, salinity fluctuated between 30 ppt and 2 ppt; D2; salinity fluctuated between 30 ppt and 10 ppt;
D3, salinity fluctuated between 30 ppt and 20 ppt; Co; salinity constant at 30 ppt for days 0-20, decreased to 2 ppt following subsample
on day 20; C, salinity constant at 30 ppt. Sample sizes are n=5 for each group. See text for details of experimental procedure.

c9

Figure 4-2. Mean plasma osmolality values measured for groups experiencing increases in salinity during the course of the experiment.
Group designations are as follows: I j, salinity fluctuated between 30 ppt and 40 ppt; I2; salinity fluctuated between 30 ppt and 50 ppt; I3,
salinity fluctuated between 30 ppt and 60 ppt; Q; salinity constant at 30 ppt for days 0-20, increased to 60 ppt following subsample on
day 20; C, salinity constant at 30 ppt. Sample sizes are n=5 for each group. See text for details of experimental procedure.

Plasma Osmotic Concentration (mOsm leg
to
to _
to
W

65
the values on day 20 significantly higher than values from day 0 and day 1 (px0.05); all
other groups exhibited the same trend, but differences were statistically non-significant.
This appears to be related to the experimental manipulation of fish, and not to the salinity
regime experienced by each group. All groups regulated plasma osmolality effectively. No
differences in regulatory ability could be seen among any of the groups tested, regardless of
the magnitude or direction of the salinity fluctuation.
Day 21
The above addresses the question of how effectively C. variegatus regulates plasma
osmolality in the face of salinity fluctuations. Transfer of individuals to either 2 or 60 ppt
following 20 days of exposure to a variety of salinity fluctuations addresses the second
question posed earlier Does past exposure to large salinity fluctuations result in improved
osmoregulatory performance, compared to animals experiencing little or no salinity
fluctuation, immediately following a fluctuation in salinity? All measurements here were
taken 24 h (on day 21) following the final salinity fluctuation.
Hematocrit results showed no consistent trends on day 21 samples. Only one
significant difference was noted, with the Hct value for group Q being significantly elevated
compared to the value for group C (p<0.05). All other comparisons showed no significant
differences in Hct.
There were significant differences in plasma osmolality for day 21 measurements.
Here groups which had experienced greater fluctuations in salinity over the course of the
experiment showed a much better ability to osmoregulate compared to the control group or
groups which had undergone small salinity fluctuations. Increasing salinity seemed to have
a greater impact on plasma osmolality than did decreasing salinity (Figures 4-1 and 4-2).
Plasma osmolality in groups Dj, [>, and D3 all exhibited declines on day 21
relative to day 20, but these differences were not statistically significant. No differences in
regulatory ability among these three groups, or with the control group (group C), could be

66
discerned. However, group Cq, which had been maintained at 30 ppt for the first 20 days of
the experiment, did show a significant decrease in plasma osmolality on day 21 relative to
values measured on days 0 through 20. Furthermore, this group had lower osmolality
values on day 21 than all other groups, although this difference was only statistically
significant when compared to groups C and D3.
More striking differences existed for groups 11, h, I3, and Q. In all cases plasma
osmolality was elevated on day 21 relative to measurements taken from the same group on
day 20 or earlier. Group I3, which had experienced daily fluctuations in salinity of 30 ppt,
showed the smallest increase in osmolality values, being significantly higher than
measurements from day 0 only. Group U showed a similar result, with the plasma
osmolality value from day 21 significantly higher than values from day 0 and day 2 only.
Groups Ii and C| showed the largest changes in plasma osmolality on day 21. Both groups
had osmolality values that were significantly higher than all previous measurements taken
from their respective groups. The value for group Ii was elevated compared to groups I2,13,
and C, although this increase was statistically significantly only when compared to group C.
Group C| was significantly elevated when compared to all other groups (including group
It).
Discussion
The ability to adjust rapidly to altered salinities would be an obvious advantage to
salt marsh organisms. Physiological responses of euryhaline fishes exposed to rapid
changes in salinity can be grouped into two phases (Holmes and Donaldson, 1969): an
adaptive period and a regulatory period. During the adaptive period, plasma osmolality
varies, gradually returning to values approaching original levels. In the regulatory period,
plasma osmolality is more finely controlled as the fish adjusts to the altered salinity and
reaches ionic homeostasis. Fishes which reach the regulatory period quickly (i.e., have short
adaptive periods) should be best able to tolerate alterations in ambient salinity. Although the

67
length of the adaptive period was not measured in the present study, results nevertheless
suggest that it is relatively short for C. variegatus.
Nordlie (1985) showed that C. variegatus was an excellent osmotic regulator over a
salinity range from 0.3 ppt to 70 ppt. Plasma osmolality values changed only slightly over
this range of salinities in his study, varying by only 40 mOsm kg'l. However, fish in his
study were fully acclimated to each experimental salinity prior to testing. The present study
indicates that C. variegatus is an excellent regulator of plasma osmolality even when fishes
are exposed to large daily fluctuations in salinity. Although fish in all groups in the present
study regulated at slightly higher levels than seen by Nordlie (1985), plasma osmolality
values varied similarly, with differences of less than 40 mOsm kg' 1 for all groups on days
0 through 20. The transfer process itself elicited much of this variation, with slight increases
in plasma osmolality seen over the time course of the experiment. A similar trend was seen
in a study by Woo and Wu (1982) on the red grouper, Epinephelus akaara, and the black
sea bream, My lio macrocephalus.
The influence of a single alteration in salinity on osmoregulatory ability has been
studied for a number of fishes, with changes in plasma osmolality observed in the present
study of similar magnitude to those seen in other curyhaline fishes (Wakeman and
Wohlschlag, 1983; Engel ct al., 1987; Ferraris et al., 1988; Mancera et al., 1993; Yoshikawa
et al., 1993; Altimiras et al., 1994; Tort et al., 1994). Few studies have examined how
fluctuating salinities influence osmoregulation of fishes. The best comparison of my results
are with a study on Blennius pholis by Davenport and Vahl (1979). In their study B. pholis
were exposed to alternating periods of freshwater and seawater, with each period lasting 6 h.
Similar to the results from the present study, plasma osmolality did not vary significantly
over the course of the experiment.
No consistent differences in Hct were observed over the course of the experiment.
Nordlie ct al., (1995) found that C. variegatus efficiently regulates water content over a
wide range of salinities, with a difference of only 4% observ ed between 0 ppt and 100 ppt.

68
These results suggest that there are no large movements of water between the blood and
tissues as a result of fluctuations in salinity over the range tested. The lack of response in
hematocrit in this study may also indicate that within the range of salinities studied C.
variegatus is subject to normal, or tolerance physiological processes, as an increase in
hematocrit values would be expected if C. variegatus were exposed to conditions leading to
resistance processes (Swift, 1982).
It has been hypothesized that fishes occurring in a habitat where salinity often
fluctuates may be able to respond more quickly and completely to alterations in salinity, and
that the limits of tolerance are farther apart if salinity fluctuates periodically (Gunter, 1967;
Spaargaren, 1974; Johnston and Cheverie, 1985). My results provide evidence that prior
exposure to fluctuations in salinity does impart an osmoregulatory advantage. Fishes
previously exposed to large fluctuations in salinity regulated plasma osmolality better than
fishes that had previously experienced no change or small changes in salinity. Increasing
salinity had the greatest impact on regulation of plasma osmolality. Group C|, which had
experienced no prior change in salinity, and group I j, exposed to the smallest fluctuations in
salinity prior to being transferred to 60 ppt showed large increases in plasma osmolality
compared to the control group and groups which had previously experienced large
fluctuations in salinity. Decreasing salinity groups exhibited the same pattern, but
differences in osmolality were less pronounced. Only group Cp, which had not been
previously exposed to fluctuations in salinity, showed a significant decline in plasma
osmolality after transfer to 2 ppt. Salinities between 2 ppt and 35-40 ppt are typically
encountered by this population of C. variegatus in its native habitat, with salinities as high
as 60 ppt rarely encountered. Thus, it is not surprising that transfer to 60 ppt elicited greater
changes in regulation of plasma osmolality.
Variations in salinity imposed on fish in this study caused no dramatic effects. No
adverse behavior or mortalities were noted throughout the course of the experiment. Thus,
despite the differences seen in regulation of plasma osmolality between some of the

69
experimental groups, these differences were not large enough in magnitude to cause
observable distress in the experimental animals. These results indicate that C. variegatus is
well adjusted for life in variable salinity conditions. It would be interesting to compare
results from the present experiments with studies examining the influence of fluctuating
salinity on osmoregulation in fishes that can tolerate changing salinity, but that experience
infrequent salinity variations in their native habitat.

CHAPTER 5
INFLUENCE OF ENVIRONMENTAL SALINITY ON BLOOD OXYGEN LEVELS OF
CYPRINODON VARIEGATUS
Introduction
The sheepshead minnow, Cyprinodon variegatus, is a euryhaline teleost whose
typical habitats are brackish water, coastal salt marshes that experience frequent salinity
fluctuations. Variations in environmental salinity may directly affect the respiratory system
of fishes in at least two ways: by affecting the solubility of oxygen in the water pumped
over the gills, and by affecting the solubility of oxygen dissolved in plasma. Changes in the
ionic composition of bodily fluids may also interact with oxygen to influence tolerance to
variable salinity conditions (Truchot, 1987). Furthermore, fishes in saline water with low
oxygen tension must balance maximizing branchial oxygen diffusion with greater
osmoregulatory demands due to the accompanying increases in ion and water exchange
(Perry and McDonald, 1993). Additionally, the oxygen content of many aquatic habitats is
subject to large natural fluctuations, so oxygen is a potentially limiting factor by itself
(Dejours, 1987; Graham, 1990). This is especially true in shallow waters, where chronic or
periodic hypoxia may be a common phenomenon (Graham, 1990). Salt marsh habitats are
often exposed to hypoxic conditions (Renaud, 1985;Toulmond, 1987).
Cyprinodon variegatus is an extremely competent euryhaline tcleosL Over a range
of salinities from freshwater (0 ppt) to 40 ppt, very small changes in metabolism occur
(Nordlic, et al., 1991; this study, Chapter 2). Both oxygen consumption and critical oxygen
tension (Pc) arc essentially unaffected by changes in salinity over this range. Salinities
above this range cause metabolic adjustments, with increases in Pc and decreases in
metabolism observed (this study, Chapter 2). The mechanisms involved in maintaining
70

71
constant metabolism and Pc at low to moderate salinities, and the alterations of metabolism
and Pc at higher salinities likely involve adjustments in oxygen transport. Not only do high
salinity waters have lowered concentrations of dissolved oxygen, but salinity itself seems to
elicit physiological responses similar to those resulting from lowered levels of oxygen.
Increasing blood oxygen levels with increasing salinity, possibly leveling off at the highest
salinities, could lead to the observed responses in metabolism and Pc in C. variegatus.
Increased blood gas transport is likely the primary mechansim used by most fishes
to increase the amount of oxygen delivered to tissues. Blood oxygen transport in most
teleosts is dependent upon the respiratory pigment hemoglobin. Blood oxygen transport is
normally increased by increasing the concentration of hemoglobin, increasing the number
of erythrocytes in circulation, and/or adjusting the affinity of hemoglobin for oxygen
(Davis, 1975; Wells et al., 1989; Jensen et al., 1993; Perry and McDonald, 1993). I
examined hemoglobin concentration, erythrocyte count, and packed cell volume (hematocrit)
over a wide range of salinities for individuals of C. variegatus to evaluate the influence of
salinity on blood oxygen levels in this extremely curyhaline species.
Methods
Collections of fish, transportation back to the laboratory, and general lab procedures
were performed as described previously. Using the same protocol described earlier, fish
were sequentially acclimated to a series of salinities ranging from 0 ppt to 80 ppt (0 to 2285
mOsm kg‘l). At the end of the acclimation period, fish were sacrificed to determine
hemoglobin concentration ([Hb]), hematocrit (Hct), and erythrocyte (RBC) count.
Blood Sampling
Fish were first carefully netted from their experimental aquaria and blotted dry.
Blood was taken by sternal cardiac puncture using freshly heparinized microhcmatocrit
tubes drawn to a fine point. Blood from each fish was collected in two microhcmatocrit

72
tubes. Once the first tube was filled with a volume > 20 (xl, the blood was immediately
dispensed into a small ceramic crucible. Eppendorf® micropipettes were then used to
dispense aliquots of blood from the crucible into test tubes used in the determination of
hemoglobin concentration and erythrocyte count. A second microhematocrit tube was then
filled and used for determination of the hematocrit, and mass and standard length of fish
were then determined. Fish were used without regard to sex, and all blood samples were
taken between 0700 h and 1100 h. Food was withheld from experimental aquaria for 24 h
prior to testing to ensure that all fish were post-absorptive. Statistical procedures followed
Winer et al. (1991) and Sokal and Rohlf (1995). All statistical analyses were one way tests
using the Tukey-Kramer post hoc comparison (p = 0.05).
Hemoglobin Analysis
Hemoglobin concentration was measured spectrophotometrically on 10 pi samples
using the cyanomethemoglobin method (Brown, 1993). As recommended by Innes and
Wells (1985), the cyanomethemoglobin solutions were centrifuged for 10 min at 5000 g
prior to colorimetric determination to remove erythrocyte debris. This method gives reliable
results when used with fish blood (Blaxhall, 1972; Blaxhall and Daisley, 1973; Coburn,
1973; Innes and Wells, 1985).
Erythrocyte Count
Erythrocytes were counted immediately following dilution in test tubes containing
Natt and Herricks solution (Campbell and Murru, 1990). This solution acts as both stain
and dilutent and is routinely used for counting erythrocytes of fish (Campbell and Murru,
1990). A 1:200 dilution was used and cells were counted in an improved Ncubauer
hcmatocytomcter following precautions outlined in Brown (1993).

73
Hematocrit
Hematocrit was measured to determine the packed cell volume of the erythrocytes
contained in the blood. Immediately after filling the second microhematocrit tube one end
was sealed and the tube placed into a micro-hematocrit centrifuge. Tubes were then
centrifuged for 10 min to separate plasma from formed elements. The hematocrit was read
using a micro-capillary reader and expressed as percent erythrocyte.
From the test values obtained, mean corpuscular volume (MCV), mean corpuscular
hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC) were
calculated for each fish as follows (Brown, 1993):
Mcv.Hc.xioy mch.WW|Mchc.EM
RBC/1 RBC/1 Hct
These erythrocyte indices are used to further define the relationship between hemoglobin
content and size of the erythrocyte.
Results
The various measures of blood oxygen are arranged by salinity group in Table 5-1.
Significant differences over the range of test salinities were found for all parameters except
MCHC. Body mass had no significant influence on any of the measured or calculated
blood oxygen indices.
Salinity exerted the greatest influence on erythrocyte count (Figure 5-1). Values
obtained at 80 ppt and 0 ppt were significantly higher than all other salinities. Erythrocyte
count was next highest at 60 ppt and 70 ppt, being significantly elevated compared to values
in fish acclimated to 2, 10, 20, and 30 ppt. Fish acclimated to salinities from 2 ppt through
50 ppt exhibited no significant differences in erythrocyte count.
Measurements of hemoglobin concentration exhibited a similar pattern, although
fewer significant differences were noted (Figure 5-2). Mean values of fishes acclimated to 0
ppt were highest and significantly different from fish acclimated to salinities from 2 ppt

Table 5-1. Hematocrit (Hct), hemoglobin concentration ([Hb]), erythrocyte count (RBC), mean corpuscular hemoglobin (MCH), mean
corpuscular volume (MCV), and mean corpuscular hemoglobin concentration (MCHC) as a function of salinity for Cyprinodon
variegatus. All values are expressed as means ± se. See text for further details on blood indices.
Salinity (ppt)
n
Hct (%)
[Hb] (gdl-l)
RBC (x 106 mm*3)
MCH (pg)
MCV (pm3)
MCHC (%)
0
9
29.56 ± 0.90
7.52 ± 0.35
2.79 ± 0.06
26.93 ± 1.08
106.23 ± 3.78
25.52 ± 1.10
2
12
20.33 ± 1.28
5.44 ± 0.22
1.91 ± 0.07
28.50 ± 0.83
105.76 ± 4.90
27.51 ± 1.49
10
10
23.80 ± 1.55
5.23 ± 0.29
1.88 ± 0.12
28.08 ± 1.37
128.51 ± 8.51
22.34 ± 1.13
20
9
19.56 ± 1.19
4.73 ± 0.23
1.71 ± 0.05
27.68 ± 1.15
113.62 ± 4.65
24.69 ± 1.38
30
10
21.10 ±0.89
5.55 ± 0.39
1.88 ± 0.14
29.80 ± 1.64
114.92 ± 5.04
26.13 ± 1.22
40
12
22.83 ± 1.22
5.85 ± 0.20
2.16 ± 0.06
27.13 ± 0.52
106.58 ± 6.13
26.20 ± 1.35
50
10
22.5 ±1.21
5.82 ± 0.35
2.06 ± 0.07
28.39 ± 1.54
109.26 ± 4.26
26.39 ± 1.87
60
10
25.2 ± 1.67
6.75 ± 0.40
2.36 ± 0.08
28.45 ± 1.08
106.34 ± 5.05
27.26 ± 1.73
70
10
25.4 ± 1.87
6.09 ±0.16
2.28 ± 0.08
26.89 ± 0.57
112.09 ± 8.30
24.88 ± 1.46
80
10
22.9 ± 1.27
6.43 ± 0.29
2.81 ± 0.08
23.04 ± 1.06
81.88 ± 4.40
28.89 ±2.12

Figure 5-1. Mean erythrocyte (RBC) count over a range of salinities in Cyprinodon variegcitus (bars indicate ± se; numerical values
above the points in the figure indicate sample size at each salinity).


Figure 5-2. Mean hemoglobin concentration ([Hb]) over a range of salinities in Cyprinodon vciriegcitus (bars indicate ± se; numerical
values above the points in the figure indicate sample size at each salinity).

Salinity (ppt)
Hemoglobin Concentration (g dl )
roGo-^tnai-sioocoo
8¿

79
through 50 ppt. Hemoglobin concentration was also elevated in fish acclimated to 60 ppt, 70
ppt, and 80 ppt, although these values were only significantly higher than the value of fish
acclimated to 20 ppt.
Hematocrit measurements showed less dependence upon salinity (Figure 5-3).
Mean hematocrit was highest in 0 ppt, and was significantly different than mean values of
fish acclimated to all salinities except 60 ppt and 70 ppt. No other significant differences in
hematocrit were noted.
Calculated erythrocyte indices indicated a slightly different pattern. The average
concentration of hemoglobin in the erythrocyte (MCHC) did not vary significantly among
salinity acclimation groups. However, both the average weight of hemoglobin in the
erythrocyte (MCH) and the average volume of the erythrocyte (MCV) were lowest in fishes
acclimated to 80 ppt. MCH was significantly depressed in fish acclimated to 80 ppt when
compared to groups acclimated to 2, 30, 60, and 70 ppt, with MCV values at 80 ppt
significantly lower than values obtained for groups at 10, 20, 30, 50, and 70 ppt.
Discussion
Changes in environmental salinity can exert profound effects on blood oxygen
transport. Increases in salinity confront fishes with the necessity of satisfying oxygen
requirements under conditions of reduced oxygen availability. Fishes may exploit multiple
strategies to optimize blood oxygen transport. The amount of oxygen delivered to the
tissues by the blood per unit time is a product of the cardiac output, the oxygen tension
difference between arterial and venous blood, and the blood oxygen capacitance coefficient
(Jensen, 1991; Jensen ct al., 1993). The capacitance coefficient reflects the hemoglobin's
oxygen transporting properties, and can be adjusted in what have been termed 'qualitative'
and 'quantitative' ways (Jensen, 1991). Regulation of hemoglobin-oxygen (Hb-CH) affinity
represents the primary method for qualitatively altering oxygen carrying capacity, with
control of hemoglobin concentration the primary quantitative mechanism (Jensen, 1991).

Figure 5-3. Mean hematocrit (Hct) over a range of salinities in Cyprinodon variegatus (bars indicate ± se; numerical values above the
points in the figure indicate sample size at each salinity).

35
30
25
20
15
10
*
\
i
\
oo
' • i 1 | i i * i j i i i i j i i i i | i i * i | i i i I | i i i i | i i i i | r i i i | i i i t
0 10 20 30 40 50 60 70 80 90
Salinity (ppt)

82
Blood oxygen carrying capacity can also be increased quantitatively by release of stored
erythrocytes, by accelerating maturation of immature erythrocytes, and/or by production of
new erythrocytes (Murad et al., 1990); release of erythrocytes from storage organs (e.g.,
spleen) appears to be the most likely scenario (Soivio et al., 1980; Wells et al., 1989). Fish
exposed to water of changing salinity would be expected to experience variability in their
blood oxygen capacitance coefficient and blood oxygen carrying capacity (Jensen et al.,
1993). Quantitative mechanisms for adjusting blood oxygen carrying capacity were
examined in this study.
Few studies have examined the influence of salinity on oxygen carrying capacity of
fishes. Guernsey and Poluhowich (1975) examined the blood oxygen capacity of American
eels (Anguilla rostrata) acclimated to 0 ppt, 24 ppt, and 34 ppt. As in C. variegatus,
hematocrit was highest in eels acclimated to 0 ppt. However, while oxygen capacity of
acclimated eels was higher in 0 ppt than 34 ppt, the highest oxygen capacity was seen in
eels acclimated to 24 ppt. In a similar study with the cichlid Oreochromis niloticus, Sun et
al., (1995), observed a similar effect of salinity on measures of blood oxygen, with
hemoglobin concentration significantly higher in 0 ppt than in higher salinities (5 to 20
ppt).
Other factors may also contribute to variations in oxygen carrying capacity of
fishes. Hall and Gray (1929) were among the first to note that there is a general correlation
between the habits of fishes and the hemoglobin concentration of their blood. More recent
studies have shown that this generalization also applies to ery throcyte count and hematocrit
(e.g., Haws and Goodnight, 1962; Cobum, 1973; Larsson et al., 1976; Putnam and Freel,
1978). In general, highly active fishes and those that regularly encounter hypoxic conditions
have elevated blood oxygen carrying capacity relative to other species. Values determined
for C. variegatus in the present study arc comparable to other fishes with similar activity
levels, and indicate that C. variegatus docs not possess exceptionally high oxygen carrying

83
capacity at any salinity tested (Hattingh, 1972; Cobum, 1973; Larsson et al., 1976; Putnam
and Freel, 1978; Smit and Hattingh, 1979; Pelster et al., 1988b).
Salinity had a significant effect on blood oxygen carrying capacity in C. variegatus.
However, differences were seen only at the very highest and lowest salinities tested. As
expected, oxygen carry ing capacity increased at high salinities. Elevations were seen at 60
ppt through 80 ppt when compared to measures made at salinities between 2 ppt and 50 ppt.
One finding of interest was indicated by MCV measurements. These results showed that at
80 ppt overall size of erythrocytes had decreased, implying the production and/or release of
smaller, possibly immature erythrocytes. One possible explanation for this is that smaller
erythrocytes have been found to be more effective in oxygen exchange than larger
erythrocytes (Cobum, 1973) due to a larger surface area per unit volume, which may allow a
faster rate of gas exchange. Energetic constraints related to viscosity of the blood may also
play a role. A balance must exist between the advantages for oxygen transport from
increased hematocrit with a disadvantage due to increased viscosity (Wells and Weber,
1991). Hematocrit began to increase as salinity reached 60 ppt, but declined back towards
normal values at 80 ppt. It is possible that at extremely high salinities even small increases
in hematocrit lead to significant viscosity problems. Utilizing smaller erythrocytes at
extreme hypersalinities may help offset this problem.
Unexpectedly, oxygen carrying capacity was highest in the group acclimated to 0
ppt, as indicated by measurements of all blood indices. Although freshwater is tolerated by
C. variegatus, previous work has shown that acclimation to freshwater is difficult and may
result in significant mortality unless decreases in salinity are made in small steps (Nordlic
and Walsh, 1989; Nordlic et al., 1991). Survival in freshwater requires many of the same
responses that arc necessary at extremely high salinities, with both freshwater and
hypcrsalinc conditions imposing difficult osmoregulatory problems for C. variegatus. In
both situations, proliferation of mitochondria rich cells on gill cpithclia is needed to
maintain ionic balance (Evans, 1984; Evans, 1993; Wood and Marshall, 1994). However,

84
this extends the surface involved in ionic exchange at the expense of gill epithelia involved
in gas exchange. Recent studies have shown that such proliferation of mitochondria rich
cells does impair respiratory gas transfer (Bindon et ah, 1994a; Bindon et al., 1994b).
Mechanisms to increase oxygen carrying capacity of the blood would be expected under
such conditions.
However, freshwater conditions differ significantly from hypersaline conditions in
several ways. Most importantly, metabolism is reduced at extreme hypersalinities (this
study, Chapter 2). In conjunction with elevated Pc, depressed metabolism at these
hypersalinities greatly reduces energetic expenditures, partially alleviating the need for
increased oxygen carrying capacity. Thus, whereas measures of blood oxygen are elevated
at salinities of 60 ppt and higher, increases were moderated by a reduction in overall
energetic expenditures. Fish acclimated to 0 ppt exhibit insignificant reductions in
metabolism, so possible increases in oxygen needs in freshwater can not be compensated
for in this manner. Furthermore, a number of studies have indicated that Hb-Oy affinity is
decreased in freshwater conditions (e.g., Benditt et al., 1941; Weber et al., 1976; Woo and
Wu, 1982). Thus, oxygen carrying capacity may have to be increased in quantitative ways if
oxygen affinity of the hemoglobin molecule is decreased in dilute media.
Another finding from the present study was that C. variegatus exhibits little change
in quantitative measures of oxygen carrying capacity over the range of salinities between 2
ppt and 50 ppt. Previous studies have shown that salinities within this range have little
effect on the metabolic rate, Pc, and osmoregulatory ability of C. variegatus (this study,
Chapters 2,3, and 4). However, as oxygen needs would be expected to rise as salinity was
increased over this range, oxygen carrying capacity was expected to increase as well.
Several possibilities may explain why blood oxygen carrying capacity did not
increase over this range of salinities. First, increases in salinity may require little
compensation in oxygen carrying capacity until extreme hypcrsalinitics arc reached.
However, it is more likely that the lack of a response may have been due to the fact that

85
large variations in salinity alone seldom occur under natural conditions. A larger response
would be expected if salinity were varied together with oxygen and/or temperature. Another
possibility is that C. variegatus, like many fish species, may utilize multiple hemoglobins.
Weber (1990) noted that the use of multiple hemoglobins can extend the range of
conditions under which oxygen is transported efficiently, thus enlarging the range of
available habitats. If C. variegatus utilizes such a mechanism, large quantitative changes in
oxygen carrying capacity might not be seen.
Finally, C. variegatus may exhibit mostly qualitative changes in response to salinity,
i.e., the primary response to changing salinity may be better reflected in Hb-C>2 affinity.
Changes in Hb-C>2 affinity as a result of environmental changes are extremely common in
fishes (e.g., Johansen and Weber, 1976; Wells et al., 1980; Weber, 1981), although this has
rarely been examined with respect to salinity. However, studies on Anguilla anguilla
(Weber et al., 1976) and Salmo salar (Máxime et al., 1990) did find that increases in
salinity between freshwater and seawater led to increases in Hb-Cb affinity. A high affinity
hemoglobin molecule might also be advantageous under hypcrsaline conditions, although it
may be ineffective during activity (McMahon, 1988). As Hb-Cb affinity was not measured
in this study, direct correlation with C. variegatus is purely speculative at this time.
Nevertheless, changes in Hb-Cb affinity would correlate well with the large decreases in
energetic expenditures exhibited by C. variegatus at high salinities. However, other factors
could also be involved. First, C. variegatus may exhibit low salt sensitivity over the normal
range of salinities encountered to reduce dependence of blood oxygen affinity on
environmental salinity (Weber et al., 1976). Second, the primary mechanism used to alter
Hb-Cb affinity results from swelling of erythrocytes (Jensen, 1991; Wells and Weber,
1991; Jensen et al., 1993). Swelling of erythrocytes is normally accompanied by a rise in
cell pH and decreased red cell hemoglobin, NTP and ATP concentrations, all of which serve
to increase the oxygen affinity (Wells and Weber, 1991). However, as seen by the MCHC
and MCV values determined in this study, ery throcytes did not swell in response to

86
increased salinity in C. variegatus. Thus if Hb-02 affinity is altered with changes in
environmental salinity, it must be changed in some other manner.
This study clearly indicates that salinity does influence the oxygen carrying capacity
of the blood of C. variegatus. Quantitative differences in hemoglobin concentration,
hematocrit, and erythrocyte count were noted in response to changing salinity. As discussed
above, it also seems likely that salinity’ may influence qualitative changes in blood oxygen
transport in C. variegatus. Further research is needed to better understand the influence of
salinity on blood oxygen levels in euryhaline teleosts.

CHAPTER 6
SUMMARY AND CONCLUSIONS
This study examined costs associated with life of a teleost in a variable salinity
environment, represented here by a salt marsh. Cyprinodon variegatus was used to examine
the influence of salinity on routine metabolic rate (RMR), critical oxygen tension (Pc),
osmoregulation, and blood oxygen carrying capacity. Results are summarized below.
1) Field measurements in the Cedar Key salt marsh indicated that this habitat undergoes
extensive variation in salinity, temperature, and oxygen.
2) RMR was relatively constant over a range of salinities from 0 ppt to 40 ppt. At higher
salinities RMR began to decline, and was significantly depressed under hypcrsaline
conditions.
3) Following sequential acclimation to experimental salinities, Pc was unaffected by
changes in salinity between 0 ppt and 40 ppt, with Pc increasing at higher salinities.
4) Reduction in metabolism and rise in Pc corresponded well with a reduced ability of C.
variegatus to regulate plasma osmolality efficiently. Osmotic permeability of the gills may
be reduced at high salinities to offset osmotic losses or ionic gains to/from the environment,
indirectly reducing the potential for oxygen uptake as well.
5) Variations in RMR and Pc as a function of environmental salinity observed in this study
suggest that C. variegatus responds to high salinities by reducing energy expenditures.
These responses effectively increases the time C. variegatus can tolerate such conditions,
albeit at a cost of a reduction in energetic processes. This strategy fits the concept of scope
for survival, as described by Hochachka (1990).
6) When C. variegatus was exposed to simulated tidal changes in salinity, RMR was
unaffected in salinity trials where both acclimation and final salinities were in the range
87

88
typically encountered by this population in its native habitat Where the acclimation or final
salinities were extremely high (50 and 60 ppt) or extremely low (0 ppt), RMR was
depressed.
7) Acclimation state was the most important factor determining the metabolic response to
simulated tidal changes in salinity. However, direction of the salinity change also influenced
metabolism in C. variegatus, with increasing salinity dealt with more efficiently than
decreasing salinity.
8) Simulated tidal experiments corroborate the hypothesis that C. variegatus tolerates
extremes in salinity by lowering metabolism, and hence decreasing energy expenditures.
Following adverse conditions metabolism returns to normal levels.
9) Cyprinodon variegatus is an excellent regulator of plasma osmolality even when
exposed to large fluctuations in salinity within the range of salinities typically encountered.
Daily fluctuations in salinity of up to 30 ppt elicited no significant differences in
osmoregulatory ability when compared to control fish.
10) Prior exposure to fluctuations in salinity does impart an osmoregulatory advantage.
Fishes previously exposed to large fluctuations in salinity regulated plasma osmolality
better than fishes that had previously experienced no or small changes in salinity.
Increasing salinity had a greater impact on regulation of plasma osmolality than did
decreases in salinity.
11) Salinity had a significant effect on blood oxygen carrying capacity in C. variegatus,
although differences were only noted at the very highest (60 to 80 ppt) and lowest (0 ppt)
salinities tested. Oxygen carry ing capacity and all blood indices were highest in the group
acclimated to 0 ppt.
12) C. variegatus exhibited little change in oxygen carrying capacity over the range of
salinities between 2 ppt and 50 ppt. Possible reasons for this include: (a) increases in
salinity may require little compensation in oxygen carrying capacity; (b) detectable changes
may only occur when salinity is varied in conjunction with variations in oxygen and/or

89
temperature; (c) C. variegatus may utilize multiple hemoglobins, and/or; (d) the primary
mechanism to increase oxygen carrying capacity may instead be through adjustment of Hb-
02 affinity.
13) Erythrocyte count was the most consistent and hematocrit the least consistent measure
of the influence of salinity on blood oxygen level.
Competition or predation pressure may be less intense in harsh, fluctuating
environments, and that certain species may avoid these pressures by evolution of wide
physicochemical tolerances and the use of such environments (Matthews and Styron,
1981). Cyprinodon variegatus seems to fit this mold well, as this species appears to be a
generalist that very successfully inhabits harsh and variable habitats where it does not have
to be very efficient to compete with other species of fishes (Martin, 1972; Berry, 1987).
This argument may explain why C. variegatus does not invade freshwater in more locales,
and why they are not very abundant in most freshwater systems; except in south Florida
freshwaters where temperature and dissolved oxygen arc variable and extreme.
Furthermore, it is believed that C. variegatus has conquered its wide geographic
range by physiological flexibility rather than by local accommodation in physiology or life
history (Berry, 1987). This hypothesis remain to be rigorously tested, although this study
lends further evidence for the physiological flexibility of C. variegatus. Examination of the
physiological ability of other populations of C. variegatus may help to resolve this issue.
The success of a fish species in a stressful environment may depend primarily
upon the proportion of the population that survives the adverse conditions, with even short¬
term increases in tolerance significant (Matthews and Styron, 1981). This hypothesis
appears to be relevant to the results of the present study. Cyprinodon variegatus is well
adapted to a varying salinity environment. Its metabolic rate, Pc, and ability to efficiently
osmoregulatc is unaffected by changes in salinity over the typical range encountered, even
when salinity is changed very rapidly. Furthermore, C. variegatus appears to tolerate
extremes in salinity by decreasing energy expenditures, waiting for conditions to improve,

90
and then responding by increasing metabolism back to normal levels. This is a potentially
adaptive response for life in a variable salinity environment.
While this study answers many questions relating to the physiology of salinity
adaptation in C. variegatus, it also raises new ones. Chief among these is the question of
how responses due to salinity would be influenced by interactions with changes in
temperature and/or dissolved oxygen. For example, C. variegatus normally experiences
high salinities together with high temperatures. The interaction of these two parameters on
osmoregulation, metabolism, and oxygen carrying capacity would be a natural continuation
of the current study. Another area of future research would include an examination into the
possible ability of C. variegatus to utilize anaerobic metabolism under varying salinity
regimes, as it was suggested by Subrahmanyam (1980) that salt marsh fishes may utilize
anaerobic metabolism more than other groups of fishes. It has also been proposed that
permeability changes of the gill may lead to reductions in metabolism at high salinities
(Nordlie et al., 1991). A histological study of the gills of C. variegatus acclimated to a wide
range of salinities to study chloride cell recruitment and hypertrophy and changes in
functional gill surface area may be used to help resolve this question. Finally, understanding
the influence of salinity on oxygen carrying capacity requires research on Hb-Ch affinity as
a function of salinity.
Salinity is a crucial physicochemical factor that exerts an important influence on
aquatic life. Cyprinodon variegatus is an extremely competent euryhaline tcleost that can
thrive in a wide variety of coastal habitats. The assertion by Davenport and Sayer (1993)
that "fish that are capable of withstanding sudden and frequent salinity changes are
generally specialized morphologically and physiologically in ways that restrict their
lifestyle" seems erroneous when fishes such as C. variegatus are considered. While few
data exist on the responses of other fishes to wide ranges in salinity, the patterns seen in C.
variegatus may represent a general pattern for fishes inhabiting variable salinity
environments.

APPENDIX 1
CRITICAL OXYGEN TENSION FIGURES
Critical oxygen tension (Pc) figures were plotted for each fish used in the study
described in chapter 2. These plots consisted of weight-adjusted oxygen consumption rates
(mg O2 h'l) plotted against oxygen tension (mm Hg) during the interval over which
oxygen consumption was calculated. Because 111 fish were used in this study, it is
impractical to show each figure. Figures A1-1, A1-2, and A1-3 are representative samples of
these figures. Generalized Pc figures were produced for each salinity group from the mean
Pc, mean routine metabolic rate (RMR), and mean slope in the conformation region. These
are shown in figure A1-4.
91

Figure Al-1. Plot indicating the calculation of the critical oxygen tension (Pc) for an individual Cyprinodon variegcitus in water at 0 ppt.

Adjusted Routine Metabolic Rate
(mg Oxygen/hr)
1.0
0.8-
0.6-
0.4-
0.2-
Pc = 62.0 mm Hg
o.o
o
20
—1—I—I—I—I—I—I—I—I 1—I—1—I—I—I—I—I—i—I—I—I 1—.—I—
40 60 80 100 120 140 160
Oxygen Tension (mm Hg)

Figure A1-2. Plot indicating the calculation of the critical oxygen tension (Pc) for an individual Cyprinodon vciriegatus in water at 50 ppt.

Adjusted Routine Metabolic Rate
(mg Oxygen/hr)
1.0
0.0 ■ t — I ■ f” 1 "I ' ■ r- I 1 » i 1 I 1 1 1 1 I 1 1 1 r—i 1 1 1 1 1 1 r—T 1
0 20 40 60 80 100 120 140 160
Oxygen Tension (mm Hg)

Figure A1-3. Plot indicating the calculation of the critical oxygen tension (Pc) for an individual Cyprinodon variegatus in water at 1(X)
ppt.

Adjusted Routine Metabolic Rate
(mg Oxygen/hr)
1.2
1.0 -
0.8 -
0.6-
0.4 -
0.2-
0.0 +
0
Pc = 76.97 mm Hg
—| t 1 1 t r j i i i 1 1 r——i | i t ■ f f——r-—1 r ; 1 » 1 | 1 1 ■ t
20 40 60 80 100 120 140 160
Oxygen Tension (mm Hg)

Figure A1-4. Generalized critical oxygen tension (Pc) plots at each salinity used in the Pc experiments. Plots were produced by using the
mean Pc, mean routine metabolic rate (RMR), and mean slope in the conformation region for each salinity group.

Oxygen Tension (mm Hg) Oxygen Tension (mm Hg)
Oxygen Consumption Rate
(mg Oxygen/hr)
oppooppo^^-»
Nwiwbisboifib^N
Oxygen Consumption Rate
(mg Oxygen/hr)
ooopoopo —
Kiwiwinscioifib^N
Oxygen Consumption Rate
(mg Oxygen/hr)
00000000;-*;-* -*
K)W^tn(j),MCobb'-*K)
Oxygen Consumption Rate
(mg Oxygen/hr)
oooooooo -*-*;-*
kiwiúióiNibobb ^N
66

Oxygen Tension (mm Hg) Oxygen Tension (mm Hg)
ponunuoD -- p-\y ojnSy
Oxygen Consumption Rate
(mg Oxygen/hr)
00000000;-* ;-*;-*
Nwiininsalob^N
Oxygen Consumption Rate
(mg Oxygen/hr)
00000000 -*;-»;-*
NWAWOlklÓoiob-N
Oxygen Consumption Rate
(mg Oxygen/hr)
OOOOOOOO;-»^ -»
rocotín CD si OD ID O -* N
Oxygen Consumption Rate
(mg Oxygen/hr)
00000000;-»;-»;-»
Nwiinijl-NlCoiob^N
001

ponunuoo — p-\ V ain8y
Oxygen Consumption Rate
(mg Oxygen/hr)
OOOOOOOO;-*;-*;-*
kiuj^wbiscntxib^K)
Oxygen Consumption Rate
(mg Oxygen/hr)
popopopp-*;-*-*
NWAwbiNibobb^N
Oxygen Consumption Rate
(mg Oxygen/hr)
pppppppp^-*-*-*
rMwjkinmkicoÍDb^w
TOI

APPENDIX 2
HELD MEASUREMENTS
Field measurements were taken in the Cedar Key area from June 1990 through June
1991. These measurements were taken at two depths (bottom and surface) for each of four
sites in the area, whenever possible. The variables measured were oxygen concentration (mg
L'l), salinity (ppt), and temperature (°C). Figure A2-1 shows the relationships between
these three variables at each site and depth over the course of the year. Oxygen
concentration was highly inversely correlated with changes in both salinity and temperature.
Salinity seems to be most highly correlated with changing oxygen levels, although salinity
and temperature appear to be linked as well, albeit with more site to site variation.
102

Figure A2-1. Oxygen concentration (mg L'l), salinity (ppt), and temperature (°C) at four
sites in the Cedar Key area taken between June 1990 and June 1991. Values given as
means, bars indicate ± se.
a) Measurements taken on the bottom at site 1; b) measurements taken on the surface at site
1; c) Measurements taken on the bottom at site 2; d) measurements taken on the surface at
site 2; e) Measurements taken on the bottom at site 3; 0 measurements taken on the surface
at site 3; g) Measurements taken on the surface at site 4.

104
Part a
—Oxygen (mg/1)
Temperature (C)
A - Salinity (ppt)
Part b
Winter

105
Part c
Oxygen (mg/1)
Temperature (C)
Salinity (ppt)
Part d
Figure A2-1 — continued

106
Part e
Oxygen (mg/1)
•” Temperature (C)
Salinity (ppt)
Part f
Figure A2-1 -- continued

107
Part g
Oxygen (mg/1)
Temperature (C)
Salinity (ppt)
Figure A2-1 -- continued

LITERATURE CITED
Able, K.W. 1976. Cleaning behavior in the Cyprinodontid fishes: Fundulus majalis,
Cyprinodon variegatus, and Lucania parva. Chesapeake Science 17:35-39.
Able, K.W. 1990. Life history patterns of New Jersey salt marsh killifishes. Bulletin of the
New Jersey Academy of Science 35:23-30.
Adam, P. 1990. Saltmarsh ecology. Cambridge University Press, Cambridge, MA. 461 p.
Ager, L.A. 1971. The fishes of Lake Okeechobee. Quarterly Journal of the Florida
Academy of Sciences 34:53-62.
Allen, J.R.L., and K. Pye. 1992. Coastal saltmarshes: their nature and importance. Pages 1-
18 in J.R.L. Allen and K. Pye, editors. Saltmarshes. Morphodynamics,
conservation and engineering significance. Cambridge University Press, Cambridge,
MA. 184 p.
Altimiras, J., S.R. Champion, M. Puigccrver, and L. Tort. 1994. Physiological responses of
the gilthead sea bream Spar us aurata to hypoosmotic shock. Comparative
Biochemistry and Physiology 108A:81-85.
Avila, I.G., L. Koldenkova, and J.C.G. Mustclicr. 1992. Cyprinodon variegatus
(Cyprinodontiformes: Cyprinodontidae), bio-regulator of mosquito larvae of Aedes
taeniorhynchus and Culex bahamensis in the Isle of Youth, Cuba. Memborias do
Instituto Oswaldo Cruz, Rio de Janeiro 87:461.
Balment, R.J., N. Hazon, and M.N. Perrott. 1987. Control of corticosteroid secretion and its
relation to osmoregulation in lower vertebrates. Pages 92-102 in R. Kirsch and B.
Lahlou, editors. Comparative physiology of environmental adaptations, Volume 1.
Adaptations to salinity and dehydration. Karger, Basel. 204 p.
Barton, M., and A.C. Barton. 1987. Effects of salinity on oxygen consumption of
Cyprinodon variegatus. Copeia 1987:230-232.
Barus, V., and E. Wohlgemuth. 1993. Study on the collection of freshwater fishes from
eastern Cuba with taxonomical notes. Folia Zoológica 42:63-76.
Battalora, M.S.J., R.D. Ellcndcr, and B.J. Martin. 1985. Gnotobiotic maintenance of
shccpshcad minnow larvae. The Progressive Fish-Culturist 47:122-125.
Beamish, F.W.H. 1964. Respiration of fishes with special emphasis on standard oxygen
consumption II. Influence of weight and temperature on respiration of several
species. Canadian Journal of Zoology 42:177-188.
108

109
Becker, K., and L. Fishelson. 1986. Standard and routine metabolic rate, critical oxygen
tension and spontaneous scope for activity of tilapias. Pages 623-628 in J.L.
Maclean, L.B. Dizon, and L.V. Hosillos, editors. The First Asian Fisheries Forum.
Asian Fisheries Society, Manila, Phillipines. 727 p.
Beitinger, T.L., and R.W. McCauley. 1990. Whole-animal physiological processes for the
assessment of stress in fishes. Journal of Great Lakes Research 16:542-575.
Benditt, E., P. Morrison, and L. Irv ing. 1941. The blood of the Atlantic salmon during
migration. Biological Bulletin 80:429-440.
Bern, H.A., and S.S. Madsen. 1992. A selective survey of the endocrine system of the
rainbow trout (Oncorhynchus mykiss) with emphasis on the hormonal regulation of
ion balance. Aquaculture 100:237-262.
Berry, W.J. 1987. Aspects of the growth and life history of the sheepshead minnow,
Cyprinodon variegatus, from Rhode Island and Florida. Ph.D Dissertation.
University of Rhode Island, Narragansett, RI. 187 p.
Bindon, S.D., J.C. Fenwick, and S.F. Perry. 1994a. Branchial chloride cell proliferation in
the rainbow trout, Oncorhynchus mykiss: implications for gas transfer. Canadian
Journal of Zoology 72:1395-1402.
Bindon, S.D., K.M. Gilmour, J.C. Fenwick, and S.F. Perry. 1994b. The effects of branchial
chloride cell proliferation on respiratory function in the rainbow trout
Oncorhynchus mykiss. Journal of Experimental Biology 197:47-63.
Blaxhall, P.C. 1972. The haematological assessment of the health of freshwater fish. A
review of selected literature. Journal of Fish Biology 4:593-604.
Blaxhall, P.C., and K.W. Daislcy. 1973. Routine haematological methods for use with fish
blood. Journal of Fish Biology 5:771-781.
Boutilier, R.G. 1990. Respiratory gas tensions in the environment. Pages 1-13 in R.G.
Boutilier, editor. Advances in comparative and environmental physiology, Volume 6.
Vertebrate gas exchange: from environment to cell. Springcr-Verlag, Berlin. 411 p.
Boutilier, R.G., G. Dobson, U. Hoeger, and D.J. Randall. 1988. Acute exposure to graded
levels of hypoxia in rainbow trout (Salmo gairdneri): metabolic and respiratory
adaptations. Respiration Physiology 71:69-82.
Brett, J.R., and T.D.D. Groves. 1979. Physiological energetics. Pages 279-352 in W.S.
Hoar, D.J. Randall, and J.R. Brett, editors. Fish physiology, Volume 8. Academic
Press, New York, NY. 786 p.
Bridges, C.R. 1988. Respiratory adaptations in intertidal fish. American Zoologist
28:79-96.
Brown, B.A. 1993. Hematology principles and procedures. Lea & Fcbigcr, Phialdclphia,
PN. 453 p.
Cameron, J.N. 1989. The respiratory physiology of animals. Oxford University Press, New
York, NY. 353 p.

110
Campbell, T., and F. Murru. 1990. An introduction to fish hematology. Compendium of
Continuing Education for the Practicing Veterinarian 12:525-532.
Cech, J.J., Jr. 1990. Respirometry'. Pages 335-362 in C.B. Schreck and P.B. Moyle,
editors. Methods for fish biology. American Fisheries Society, Bethesda, MD.
684 p.
Chung, K.S. 1982. Salinity tolerance of tropical salt-marsh fish of Los Patos Lagoon,
Venezuela. Bulletin of the Japanese Society of Scientific Fisheries 48:873.
Cobum, C.B., Jr. 1973. Red blood cell hematology of fishes: a critique of techniques and a
compilation of published data. Journal of Marine Science 2:37-58.
Congleton, J.L. 1980. Observations on the responses of some southern California tidepool
fishes to nocturnal hypoxic stress. Comparative Biochemistry and Physiology
66A:719-722.
Conover, D.O., and S.B. DeMond. 1991. Absence of temperature-dependent sex
determination in northern populations of two cyprinodontid fishes. Canadian
Journal of Zoology 69:530-533.
Cooper, A.W. 1974. Salt marshes. Pages 55-98 in H.T. Odum, B.J. Copeland, and E.A.
McMahon, editors. Coastal ecological systems of the United States, Volume 2. The
Conservation Foundation, Washington D.C.
Darling, J.D.S. 1976. Electrophoretic variation in Cyprinodon variegatus and systcmatics
of some fishes of the subfamily Cyprinodontinae. Ph.D Dissertation. Yale
University, Ithica, NJ. 153 p.
Davenport, J., and M.D.J. Sayer. 1993. Physiological determinants of distribution in fish.
Journal of Fish Biology 43:121-145.
Davenport, J., and O. Vahl. 1979. Responses of the fish Blennius pholis to fluctuating
salinities. Marine Ecology Progressive Scries 1:101-107.
Davis, J.C. 1975. Minimal dissolved oxygen requirements of aquatic life with emphasis on
Canadian species: a review. Journal of the Fisheries Research Board of Canada
32:2295-2332.
De Vlaming, V.L., A. Kuris, and F.R.J. Parker, Jr. 1978. Seasonal variation of reproduction
and lipid reserves in some subtropical Cyprinodontids. Transactions of the
American Fisheries Society 107:464-472.
Dcjours, P. 1987. Water and air physical characteristics and their physiological
consequences. Pages 3-11 in P. Dcjours, L. Bolis, C.R. Taylor, and E.R. Wcibcl,
editors. Comparative physiology; life in water and on land, Fidia research scries,
Volume 9. Liviana Press, Padova. 556 p.
Doll, J.M., and T.F. Bast. 1969. Three estuarine killifish as fresh water mosquito larvivorcs.
Mosquito News 29:365-367.

Ill
Donnelly, J., and J.J. Torres. 1988. Oxygen consumption of midwater fishes and
crustaceans from the eastern Gulf of Mexico. Marine Biology 97:483-494.
Duggins, C.F., Jr., A.A. Karlin, and K.G. Relyea. 1983. Electrophoretic comparison of
Cyprinodon variegatus Lacépede and Cyprinodon hubbsi Carr, with comments on
the genus Cyprinodon (Atheriniformes: Cyprinodontidae). Northeast Gulf Science
6:99-107.
Dunson, W.A., P. Fricano, and W.J. Sadinski. 1993. Variation in tolerance to abiotic
stresses among sympatric salt marsh fish. Wetlands 13:16-24.
Dunson, W.A., and J. Travis. 1994. Patterns in the evolution of physiological specialization
in salt-marsh animals. Estuaries 17:102-110.
Dwyer, M., and A. Beulig. 1991. Social experience and the development of aggressive
behavior in the pupfish (Cyprinodon variegatus). Journal of Comparative
Psychology 105:398-404.
Echelle, A.A., and P.J. Connor. 1989. Rapid, geographically extensive genetic introgression
after secondary contact between two pupfish species (Cyprinodon,
Cyprinodontidae). Evolution 43:717-727.
Echelle, A.A., and A.F. Echelle. 1987. Evolutionary relationships among inland pupfishcs
of the Cyprinodon variegatus complex. Page 50 in E.P. Pister, editor. Proceedings
of the Desert Fishes Council, Volume 19. Desert Fishes Council, Las Vegas, NV.
Echelle, A.F., and A.A. Echelle. 1994. Assessment of genetic introgression between two
pupfish species, Cyprinodon elegans and C. variegatus (Cyprinodontidae), after
more than 20 years of secondary contact. Copcia 1994:590-597.
Eddy, F.B. 1982. Osmotic and ionic regulation in captive fish with particular reference to
salmonids. Comparative Biochemistry and Physiology 73B: 125-141.
Elder, J.F., Jr., and B.J. Turner. 1994. Concerted evolution at the population level: pupfish
Hindlll satellite DNA sequences. Proceedings of the National Academy of Science
91:994-998.
Engel, D.W., W.F. Hettler, L. Coston-Clements, and D.E. Hoss. 1987. The effect of abrupt
salinity changes on the osmoregulatory abilities of the Atlantic menhaden
Brevoortia tyrannus. Comparative Biochemistry and Physiology 86A:723-727.
Evans, D.H. 1984. The roles of gill permeability and transport mechanisms in curyhalinity.
Pages 239-283 in W.S. Hoar and D.J. Randall, editors. Fish physiology, Volume
10B. Academic Press, London. 416 p.
Evans, D.H. 1993. Osmotic and ionic regulation. Pages 315-341 in D.H. Evans, editor.
The physiology of fishes. CRC Press, Boca Raton, FL. 592 p.
Fcbry, R., and P. Lutz. 1987. Energy partitioning in fish: The activity-related cost of
osmoregulation in a curyhaline cichlid. Journal of Experimental Biology 128:63-85.

112
Ferraris, R.P., J.M. Almendras, and A.P. Jazul. 1988. Changes in plasma osmolality and
chloride concentration during abrupt transfer of milkfish (Chanos chanos) from
seawater to different test salinities. Aquaculture 70:145-157.
Fontaine, Y.A. 1993. Adaptations versus accommodations: some neuroendocrine aspects in
teleost fish. Fish Physiology and Biochemistry 11:147-154.
Foskett, J.K. 1987. The chloride cell. Pages 83-91 in R. Kirsch and B. Lahlou, editors.
Comparative physiology of environmental adaptations, Volume 1. Adaptations to
salinity and dehydration. Karger, Basel. 204p.
Fry, F.E.J. 1947. Effects of the environment on animal activity. University of Toronto
Studies Biological Series 55:3-62.
Fry, F.E.J. 1957. The aquatic respiration of fish. Pages 1-63 in M.E. Brown, editor. The
physiology of fishes. Academic Press Inc., New York, NY. 447 p.
Furspan, P., H.D. Prange, and L. Greenwald. 1984. Energetics and osmoregulation in the
catfish, Ictalurus nebulosus and /. Punctatus. Comparative Biochemistry and
Physiology 77A:773-778.
Fyfe, J. 1985. Trophic variations in salt marsh fishes. Florida Scientist 48:28.
Gill, H.S., and I.C. Potter. 1993. Spatial segregation amongst goby species within an
Australian estuary, with a comparison of the diets and salinity tolerances of the two
most abundant species. Marine Biology 117:515-526.
Goolish, E.M., and R.S. Burton. 1988. Exposure to fluctuating salinity enhances free amino
acid accumulation in Tigriopus californicus (Copcpoda). Journal of Comparative
Physiology 158B:99-105.
Graham, J.B. 1990. Ecological, evolutionary, and physical factors influencing aquatic
animal respiration. American Zoologist 30:137-146.
Guernsey, D.L., and J.J. Poluhowich. 1975. Blood oxygen capacities of eels acclimated to
fresh-, brackish- and salt-water environments. Comparative Biochemistry and
Physiology 52A:313-316.
Gunter, G. 1967. Vertebrates in hypcrsaline waters. Contributions in Marine Science
12:230-241.
Hall, F.G. 1929. The influence of varying oxygen tension upon the rate of oxygen
consumption in marine fishes. American Journal of Physiology 88:212-218.
Hall, F.G., and I.E. Gray. 1929. The hemoglobin concentration of the blood of marine
fishes. Journal of Biological Chemistry 81:589-594.
Harrington, R.W., Jr., and E.S. Harrington. 1982. Effects on fishes and their forage
organisms of impounding a Florida salt marsh to prevent breeding by salt marsh
mosquitos. Bulletin of Marine Science 32:523-531.
Hattingh, J. 1972. Observations on the blood physiology of five South African freshwater
fish. Journal of Fish Biology 4:555-563.

113
Hawkins, W.E., R.M. Overstreet, and M.J. Provancha. 1984. Effects of space shuttle
exhaust plumes on gills of some estuarine fishes: a light and electron microscopic
study. Gulf Research Reports 7:297-309.
Haws, T.G., and C.J. Goodnight. 1962. Some aspects of the hematology of two species of
catfish in relation to their habitats. Physiological Zoology 35:8-17.
Hirano, T., T. Ogasawara, J.P. Bolton, N.L. Collie, S. Hasegawa, and M. Iwata. 1987.
Osmoregulatory role of prolactin in lower vertebrates. Pages 112-124 in R. Kirsch
and B. Lahlou, editors. Comparative physiology of environmental adaptations,
Volume 1. Adaptations to salinity and dehydration. Karger, Basel. 204p.
Hochachka, P.W. 1990. Scope for survival: a conceptual "mirror" to Fry's scope for
activity. Transactions of the American Fisheries Society 119:622-628.
Holmes, W.N., and E.M. Donaldson. 1969. The body compartments and the distribution of
electrolytes. Pages 1-79 in W.S. Hoar and D.J. Randall, editors. Fish physiology,
Volume 1. Academic Press, New York, NY. 465 p.
Hughes, G.M. 1964. Fish respiratory homeostasis. Symposia of the Society for
Experimental Biology 18:81-107.
Innes, A.J., and R.M.G. Wells. 1985. Respiration and oxygen transport functions of the
blood from an intertidal fish, Helcogramma medium (Tripterygiidae).
Environmental Biology of Fishes 14:213-226.
Itzkowitz, M. 1974. The effects of other fish on the reproductive behavior of the male
Cyprinodon variegatus (Pisces: Cyprinodontidae). Behavior 48:1-22.
Itzkowitz, M. 1978. Female mate choice in the pupfish, Cyprinodon variegatus. Behavior
Proceedings 3:1-8.
Itzkowitz, M. 1981. The relationships of intrusions and attacks to territory size and quality
in the pupfish, Cyprinodon variegatus Lacépede. Biology of Behaviour 6:273-280.
Jenkins, D.K. 1981. Corticosteroids and osmoregulation in Atlantic salmon, Salmo salar
L., subjected to freshwater and seawater environments. Ph.D Dissertation. Acadia
University, Wolfville, Nova Scotia, Canada 107 p.
Jensen, F.B. 1991. Multiple strategics in oxygen and carbon dioxide transport by
haemoglobin. Pages 55-78 in A.J. Woakes, M.K. Grieshaber, and C.R. Bridges,
editors. Physiological strategies for gas exchange and metabolism. Cambridge
University Press, Cambridge, MA. 266 p.
Jensen, F.B., M. Nikinmaa, and R.E Weber. 1993. Environmental perturbations of oxygen
transport in tclcost fishes: causes, consequences and compensations. Pages 161-179
in J.C. Rankin and F.B. Jensen, editors. Fish ecophysiology. Chapman and Hall,
London. 421 p.
Job, S.V. 1969. The respiratory metabolism of Tilapia mossambica (Tclcostci). II. The
effect of size, temperature, salinity and partial pressure of oxygen. Marine Biology
3:222-226.

114
Johansen, K., and R.E. Weber. 1976. On the adaptability of hemoglobin function to
environmental conditions. Pages 219-234 in P. Spencer Davies, editor. Perspectives
in experimental biology, Volume 1. Zoology. Pergamon Press. Oxford.
Johnson, W.E. 1974. Morphological variation and local distribution of Cyprinodon
variegatus in Florida. Ph.D Dissertation. Florida Technological University,
Orlando, FL. 102 p.
Johnston, C.E., and J.C. Cheverie. 1985. Comparative analysis of ionoregulation in rainbow
trout (Salmo gairdneri) of different sizes following rapid and slow salinity
adaptation. Canadian Journal of Fisheries and Aquatic Science 42:1994-2003.
Johnston, I.A., and J. Battram. 1993. Feeding energetics and metabolism in demersal fish
species from Antarctic, temperate and tropical environments. Marine Biology
115:7-14.
Kamaky, K.J., Jr. 1986. Structure and function of the chloride cell of Fundulus heteroclitus
and other teleosts. American Zoologist 26:209-224.
Kamaky, K.J., Jr., S.A. Ernst, and C.W. Philpott. 1976. Teleost chloride cell 1. Response of
pupfish Cyprinodon variegatus gill Na/K ATPase and chloride cell fine structure to
various high salinity environments. Journal of Cell Biology 70:144-156.
Kilby, J.D. 1955. The fishes of two gulf coastal marsh areas of Florida. Tulane Studies in
Zoology 2:175-247.
Kinne, O. 1962. Irreversible nongenetic adaptation. Comparative Biochemistry and
Physiology 5:265-282.
Kinne, O. 1966. Physiological aspects of animal life in estuaries with special reference to
salinity. Netherlands Journal of Sea Research 3:222-244.
Kinne, O. 1967. Physiology of estuarine organisms with special reference to salinity and
temperature: general aspects. Pages 525-540, in G.H. Lauff, editor. Estuaries,
Volume 83. AAAS, Washington D.C. 757 p.
Kirschner, L.B. 1993. The energetics of osmotic regulation in ureotclic and hypoosmotic
fishes. The Journal of Experimental Zoology 267:19-26.
Kodric-Brown, A. 1987. Variable breeding systems in pupfishes (Genus Cyprinodon):
adaptations to changing environments. Pages 205-235 in R.J. Naiman and D.L.
Soltz, editors. Fishes in North American deserts. John Wiley and Sons, New York
NY. 552 p.
Kodric-Brown, A. 1989. Genetic introgression after secondary contact. Trends in Ecology
and Evolution 4:329-330.
Kristcnscn, K., and E. Skadhaugc. 1974. Row along the gut and intestinal absorption of
salt and water in euryhalinc teleosts: a theoretical analysis. Journal of Experimental
Biology 60:557-566.

115
Kültz, D., and H. Onken. 1993. Long-term acclimation of the teleost Oreochromis
mossambicus to various salinities: two different strategies in mastering hypertonic
stress. Marine Biology 117:527-533.
Kushlan, J.A. 1980. Population fluctuations of everglades fishes. Copeia 1980:870-874.
Larsson, A., M.l. Johansson-Sjobeck, and R. Fánge. 1976. Comparative study of some
haematological and biochemical blood parameters in fishes from the Skagerrak.
Journal of Fish Biology 9:425-440.
Leloup, J., and J.M. Lebel. 1993. Triiodothyronine is necessary for the action of growth
hormone in acclimation to seawater of brown trout (Salmo trutta) and rainbow trout
(Oncorhynchus rnykiss). Fish Physiology and Biochemistry 11:165-173.
Linton, T.K. 1992. Salinity and temperature effects on the chronic toxicity of 2,4-
dinitrophenol and 4-nitrophenol to sheepshead minnows (Cyprinodon variegatus).
Masters Thesis. The University of West Florida, Pensacola, FL. 85 p.
Lotan, R., and E. Skadhauge. 1972. Intestinal salt and water transport in a euryhaline teleost,
Aphanius dispar (Cyprinodontidae). Comparative Biochemistry and Physiology
42A:303-310.
Lowe, C.H., D.S. Hinds, and E.A. Halpem. 1967. Experimental catastrophic selection and
tolerances to low oxygen concentration in native Arizona freshwater fishes. Ecology
48:1013-1017.
Madan Mohan Rao, G. 1968. Oxygen consumption of rainbow trout (Salmo gairdneri) in
relation to activity and salinity. Canadian Journal of Zoology 46:781-786.
Madan Mohon Rao, G. 1974. Influence of activity and salinity on the weight-dependent
oxygen consumption of the rainbow trout Salmo gairdneri. Marine Biology
8:205-212.
Mancera, J.M., J.M. Perez-Figares, and P. Femandcz-Liebrez. 1993. Osmoregulatory
responses to abrupt salinity changes in the euryhaline gilthead sea bream (Sparus
aurata L.). Comparative Biochemistry and Physiology 106A:245-250.
Martin, F.D. 1968. Intraspecific variation in osmotic abilities of Cyprinodon variegatus
Laccpcde from the Texas Coast. Ecology 49:1186-1188.
Martin, F.D. 1970. Feeding habits of Cyprinodon variegatus (Cyprinodontidae) from the
Texas coast. Southwestern Naturalist 14:368-369.
Martin, F.D. 1972. Factors influencing local distribution of Cyprinodon variegatus (Pisces:
Cyprinodontidae). Transactions of the American Fisheries Society 101:89-93.
Matthews, W.J., and J.T. Styron, Jr. 1981. Tolerance of headwater vs. mainstream fishes
for abrupt physicochemical changes. American Midland Naturalist 105:149-158.
Máxime, V., M. Pcyraud-Waitzencggcr, G. Claircaux, and C. Pcyraud. 1990. Effects of
rapid transfer from sea water to fresh water on respiratory variables, blood acid-base
status and 0> affinity of haemoglobin in Atlantic salmon (Salmo salar L.). Journal
of Comparative Physiology 160B:31-39.

116
McClusky, D.S. 1989. Estuarine ecosystems. Chapman and Hall. Glasgow. 215 p.
McCormick, S.D. 1994. Ontogeny and evolution of salinity tolerance in anadromous
salmonids: hormones and heterochrony. Estuaries 17:26-33.
McMahon, B.R. 1988. Physiological responses to oxygen depletion in intertidal animals.
American Zoologist 28:39-53.
Mettee, M.F., Jr., and E.C. Beckham III. 1978. Notes on the breeding behavior, embryology
and larval development of Cyprinodon variegatus Lacépéde in aquaria. Tulane
Studies in Zoology and Botany 20:137-148.
Miller, R.R. 1981. Coevolution of deserts and pupfishes (Genus Cyprinodon) in the
American southwest. Pages 39-94 in R.J. Naiman and D.L. Soltz, editors. Fishes in
North American deserts. John Wiley and Sons, New York, NY. 552 p.
Morgan, J.D., and G.K. Iwama. 1991. Effects of salinity on growth, metabolism, and ion
regulation in juvenile rainbow and steelhead trout (Oncorhynchus mykiss) and fall
chinook salmon (Oncorhynchus tshawytschd). Canadian Journal of Fisheries and
Aquatic Science 48:2083-2094.
Moser, M.L., and L.R. Gerry. 1989. Differential effects of salinity changes on two
estuarine fishes, Leiostomus xanthurus and Micropogonias undulatus. Estuaries
12:35-41.
Moser, M.L., and J.M. Miller. 1994. Effects of salinity fluctuation on routine metabolism
of juvenile spot, Leiostomus xanthurus. Journal of Fish Biology 45:335-340.
Muir, B.S., and A.J. Niimi. 1972. Oxygen consumption of the curyhaline fish aholehole
(Kuhlia sandvicensis) with reference to salinity, swimming, and food consumption.
Journal of the Fisheries Research Board of Canada 29:67-77.
Murad, A., A.H. Houston, and L. Samson. 1990. Haematological response to reduced
oxygen-carrying capacity, increased temperature and hypoxia in goldfish, Carassius
auratus L. Journal of Fish Biology 36:289-305.
Naiman, R.J., S.D. Gerking, and R.E. Stuart. 1976. Osmoregulation in the Death Valley
pupfish Cyprinodon milleri (Pisces: Cyprinodontidae). Copcia 1976:807-810.
Naughton, S.P., and C.H. Saloman. 1978. Fishes of the nearshore zone of St. Andrew Bay,
Florida, and adjacent coast. Northeast Gulf Science 2:43-55.
Nelson, D.M. (Editor) 1992. Distribution and abundance of fishes and invertebrates in Gulf
of Mexico Estuaries, Volume 1: data summaries. ELMR Report No. 10,
NOAA/NOS Strategic Environmental Assessments Division, Rockville, MD. 273 p.
Nonnottc, G., V. Máxime, J.P. Truchot, P. Williot, and C. Pcyraud. 1993. Respiratory
responses to progressive ambient hypoxia in the sturgeon, Acipenser baeri.
Respiration Physiology 91:71-82.

117
Nordlie, F.G. 1978. The influence of environmental salinity on respiratory oxygen demands
in the euryhaline teleost, Ambassis interrupta Bleeker. Comparative Biochemistry'
and Physiology 59A:271-274.
Nordlie, F.G. 1985. Osmotic regulation in the sheepshead minnow Cyprinodon variegatus
Lacépede. Journal of Fish Biology 26:161-170.
Nordlie, F.G. 1987. Plasma osmotic Na+ and Cl' regulation under euryhaline conditions in
Cyprinodon variegatus Lacépede. Comparative Biochemistry and Phvsiology
86 A: 57-61.
Nordlie, F.G., W.A. Wahl II, J. Binello, and D.C. Haney. 1995. Body water content in the
sheepshead minnow, Cyprinodon variegatus Lacépede, over a wide range of
ambient salinities. Journal of Fish Biology 47:624-630.
Nordlie, F.G., and S.J. Walsh. 1989. Adaptive radiation in osmotic regulatory patterns
among three species of cyprinodontids (Teleostei: Atherinomorpha). Physiological
Zoology 62:1203-1218.
Nordlie, F.G., S.J. Walsh, D.C. Haney, and T.F. Nordlie. 1991. The influence of ambient
salinity on routine metabolism in the teleost Cyprinodon variegatus Lacépede.
Journal of Fish Biology 38:115-122.
Odum, H.T., and D.K. Caldwell. 1955. Fish respiration in the natural oxygen gradient of an
anaerobic spring in Florida. Copeia 1955:104-106.
Oikawa, S., Y. Itazawa, and M. Gotoh. 1991. Ontogenetic change in the relationship
between metabolic rate and body mass in a sea bream Pagrus major (Temminck &
Schlegel). Journal of Fish Biology 38:483-496.
Ott, M.E., N. Heisler, and G.R. Ultsch. 1980. A re-evaluation of the relationship between
temperature and the critical oxygen tension in freshwater fishes. Comparative
Biochemistry and Physiology 67A:337-340.
Palacios, C.A.M., and L.G. Ross. 1986. The effects of temperature, body weight and
hypoxia on the oxygen consumption of the Mexican mojarra, Cichlasoma
urophthalmus (Giinther). Aquaculture and Fisheries Management 17:243-248.
Parenti, L.R. 1981. A phylogenetic and biogeographic analysis of Cyprinodontiform fishes
(Teleostei, Atherinomorpha). Bulletin of the American Museum of Natural History
168:335-557.
Parker, A., and I. Komfield. 1995. Molecular perspective on evolution and zoogeography of
Cyprinodontid killifishes (Teleostei; Atherinomorpha). Copeia 1995:8-21.
Parvathcswararao, V. 1970. Adaptation to osmotic stress in fishes. Indian Biologist
2:16-36.
Pclstcr, B., C.R. Bridges, and M.K. Gricshabcr. 1988a. Respiratory adaptations of the
burrowing marine teleost Lunipenus lampretaeformis (Walbaum). II. Metabolic
adaptations. Journal of Experimental Marine Biology and Ecology 124:43-55.

118
Pelster, B., C.R. Bridges, A.C. Taylor, S. Morris, and R.J.A. Atkinson. 1988b. Respirator)’
adaptations of the burrowing marine teleost Lumpenus lanipretaeformis
(Walbaum). I. Ot and CO) transport, acid-base balance: a comparison with Cepola
rubescens L. Journal of Experimental Marine Biology and Ecology 124:31-42.
Perry, S.E, and G. McDonald. 1993. Gas exchange. Pages 251-278 in D.H. Evans, editor.
The physiology of fishes. CRC Press, Boca Raton, FL. 592 p.
Peterson, M.S. 1990. Hypoxia-induced physiological changes in two mangrove swamp
fishes: sheepshead minnow, Cyprinodon variegatus Lacépede and sailfin molly,
Poecilia latipinna (Lesueur). Comparative Biochemistry and Physiology
97A: 17-21.
Peterson, M.S., and R.G. Gilmore, Jr. 1988. Hematocrit, osmolality, and ion concentration
in fishes: consideration of circadian patterns in the experimental design. Journal of
Experimental Marine Biology and Ecology 121:73-78.
Pisam, M., and A. Rambourg. 1991. Mitochondria-rich cells in the gill epithelium of teleost
fishes: an ultrastructural approach. International Journal of Cytology 130:191-231.
Price, E.E., M.J. Donahue, K.L. Dickson, and J.H. Rodgers Jr. 1990. Effects of elevated
calcium concentration on Na-K-ATPase activity of two euryhaline species,
Cyprinodon variegatus and Mysidopsis bahia. Bulletin of Environmental
Contamination and Toxicology 44:121-128.
Prosser, C.L. 1955. Physiological variation in animals. Biological Review 30:229-262.
Prosser, C.L. 1975. Physiological adaptations in animals. Pages 3-18 in EJ. Vembcrg,
editor. Physiological adaptations to the environment. Thomas Y. Crovell Co. Inc.,
New York, NY. 576 p.
Putnam, R.W., and R.W. Fred. 1978. Hematological parameters of five species of marine
fishes. Comparative Biochemistry and Physiology 61A:585-588.
Quinn, T., and D.E. Schneider. 1991. Respiration of the teleost fish Anunodytes hexapterus
in relation to its burrowing behavior. Comparative Biochemistry and Physiology
98A:71-75.
Raney, EC., R.H. Backus, R.W. Crawford, and C.R. Robins. 1953. Reproductive behavior
in Cyprinodon variegatus Lacépóde, in Florida. Zoológica 38:97-103.
Rantin, F.T., A.L. Kalinin, M.L. Glass, and M.N. Fernandes. 1992. Respiratory responses
to hypoxia in relation to mode of life of two erythrinid species (Hoplias
malabaricus and Hoplias lacerdae). Journal of Fish Biology 41:805-812.
Relyca, K. 1975. The distribution of the oviparous killifishes of Florida. Science of Biology
Journal 1:49-52.
Remane, A., and C. Schlicpcr. 1971. Biology of brackish water. Wiley Intcrscicncc
Division, John Wiley and Sons, New York, NY. 372 p.

119
Renaud, M.L. 1985. Annotated bibliography on hypoxia and its effects on marine life, with
emphasis on the Gulf of Mexico. U.S. Department of Commerce, National Oceanic
and Atmospheric Administration, National Marine Fisheries Service, NOAA
Technical Report NMFS 21.
Ross, B., and L.G. Ross. 1983. The oxygen requirements of Oreochromis niloticus under
adverse conditions. Pages 134-143 in L. Fishelson and Z. Yaron, editors.
International Symposium on Tilapia in Aquaculture, Nazareth, Israel.
Ross, S.T., and T.A. Doherty. 1994. Short-term persistence and stability of barrier island
fish assemblages. Estuarine, Coastal and Shelf Science 38:49-67.
Sakamoto, T., S.D. McCormick, and T. Hirano. 1993. Osmoregulatory actions of growth
hormone and its mode of actions in salmonids: a review. Fish Physiology and
Biochemistry 11:155-164.
Schimmel, S.C., and D.J. Hansen. 1975. Sheepshead minnow (Cyprinodon variegatus): an
estuarine fish suitable for chronic (entire life-cycle) bioassays. Proceedings of the
Annual Conference of Southeastern Association of Game and Fish Commission
28:392-398.
Schwartz, F.J., G. Safrit, C. Jensen, and J. Purifoy. 1990. Stability and persistence of the
fishes inhabiting mullet pond, Shackleford Banks, North Carolina 1903-1989. The
Journal of the Elisha Mitchell Scientific Society 106:38-50.
Shipley, F.S. 1991. Oil field-produced brines in a coastal stream: water quality and fish
community recovery following long term impacts. The Texas Journal of Science
43:51-64.
Shusmin, A.G. 1989. Effects of changes of salinity on survival, oxygen threshold, and level
of standard metabolism of young of the golden mullet, Liza aurata. Voprosy
Ikhtiologii 29:1037-1040.
Simpson, D.G., and G. Gunter. 1956. Notes on habitats, systematic characters and life
histories of Texas salt water Cyprinodon tes. T ulane Studies in Zoology 4:115-134.
Skadhauge, E. 1974. Coupling of transmural flows of NaCl and water in the intestine of the
eel (Anguilla anguilla). Journal of Experimental Biology 60:535-546.
Smit, G.L., and J. Hattingh. 1979. Haematological studies on some freshwater tcleosts.
South Africa Journal of Animal Science 9:65-68.
Smit, H. 1980. Some aspects of environmental (phenotypic) adaptations in fishes.
Netherlands Journal of Zoology 30:179-207.
Soivio, A., M. Nikinmaa, and K. Wcstman. 1980. The blood oxygen binding properties of
hypoxic Salmo gairdneri. Journal of Comparative Physiology 136B:83-87.
Sokal, R.R., and F.J. Rohlf. 1995. Biometry: the principles and practice of ststistics. W.H.
Freeman and Company. New York, NY. 887 p.

120
Spaargaren, D.H. 1974. A study on the adaptation of marine organisms to changing
salinities with special reference to the shore crab, Carcinus maenas (L.).
Comparative Biochemistry and Physiology 47A:499-512.
Stout, J.P. 1985. The ecology of irregularly flooded salt marshes of the northeastern Gulf
of Mexico: a community profile. United States Fish and Wildlife Service Biological
Report 85(7.1). 98 p.
Strawn, K., and J.E. Dunn. 1967. Resistance of Texas salt- and freshwater-marsh fishes to
heat death at various salinities. Texas Journal of Science 19:57-76.
Stuenkel, E.L., and S.D. Hillyard. 1981. The effects of temperature and salinity acclimation
on metabolic rate and osmoregulation in the pupfish Cyprinodon salinus. Copeia
1981:411-417.
Subrahmanyam, C.B. 1980. Oxygen consumption of estuarine fish in relation to external
oxygen tension. Comparative Biochemistry and Physiology 67A: 129-133.
Subrahmanyam, C.B., and C.L. Coultas. 1980. Studies on the animal communities in two
north Florida salt marshes, Part III. Seasonal fluctuations of fish and
macroinvertebrates. Bulletin of Marine Science 30:790-818.
Sun, L., G. Chen, and C. Chang. 1995. Acute responses of blood parameters and comatose
effects in salt-acclimated tilapias exposed to low temperatures. Journal of Thermal
Biology 20:299-306.
Swanson, C. 1991. Aspects of the physiology of salinity adaptation in the milkfish, Chanos
chonos Forskel, during two life history stages. Ph.D Dissertation. University of
California at Los Angeles, Los Angeles, CA. 132 p.
Swift, D.J. 1982. Changes in selected blood component values of rainbow trout, Salmo
gairdneri Richardson, following the blocking of the cortisol stress response with
betamethasone and subsequent exposure to phenol or hypoxia. Journal of Fish
Biology 21:269-277.
Takei, Y. 1993. Role of peptide hormones in fish osmoregulation. Pages 136-159 in J.C.
Rankin and F.B. Jensen, editors. Fish ecophysiology. Chapman and Hall, London.
421 p.
Tang, P.S. 1933. On the rate of oxygen consumption by tissues and lower organisms as a
function of oxygen tension. Quarterly Reviews of Biology 8:260-274.
Tort, L., P. Landri, and J. Altimiras. 1994. Physiological and metabolic changes of sea
bream Sparus aurata to short-term acclimation at low salinity. Comparative
Biochemistry and Physiology 108A:75-80.
Toulmond, A. 1987. Adaptations to extreme environmental hypoxia in water breathers.
Pages 123-136 in P. Dcjours, editor. Comparative physiology of environmental
adaptations, Volume 2. Adaptations to extreme environments. Kargcr, Basel. 224 p.
Truchot, J.P. 1987. Adaptations to extreme salinities. Pages 196-207 in P. Dcjours, editor.
Comparative physiology of environmental adaptations, Volume 2. Adaptations to
extreme environments. Kargcr, Basel. 224 p.

121
Truesdale, G.A., A.L. Downing, and G.F. Lowden. 1955. The solubility of oxygen in pure
water and sea water. Journal of Applied Chemistry 5:53-63.
Turner, B.J., and R.K. Liu. 1977. Extensive interspecific genetic compatability in the new
world killifish genus Cyprinodon. Copeia 1977:259-269.
Ultsch, G.R., H. Boschung, and M.J. Ross. 1978. Metabolism, critical oxygen tension, and
habitat selection in darters (Etheostoma). Ecology 59:99-107.
Ventrella, V., F. Trombetti, A. Pagliarani, G. Trigari, M. Pirini, and A.R. Borgatti. 1992.
Salinity dependence of the ouabain-insensitive Mg“+-dependent Na+ -ATPase in
gills of rainbow trout (Oncorhynchus mykiss Walbaum) adapted to fresh and
brackish water. Comparative Biochemistry and Physiology 101B:l-7.
Verheyen, E., R. Blust, and W. Decleir. 1994. Metabolic rate, hypoxia tolerance and aquatic
surface respiration of some lacustrine and riverine African fishes (Pisces:
Cichlidae). Comparative Biochemistry and Physiology 107A:403-411.
Vemberg, W.B. 1983. Responses to estuarine stress. Pages 43-63 in B.H. Ketchum,
editor. Ecosystems of the world. Estuaries and enclosed seas. Elsevier Scientific
Publishing Company, Amsterdam. 500 p.
Von Oertzen, J.A. 1984. Influence of steady-state and fluctuating salinities on the oxygen
consumption and activity of some brackish water shrimps and fishes. Journal of
Experimental Marine Biology and Ecology 80:29-46.
Wakeman, J.M., and D.E. Wohlschlag. 1983. Time course of osmotic adaptation with
respect to blood serum osmolality and oxygen uptake in the euryhaline teleost,
Sciaenops ocellatus (red drum). Contributions in Marine Science 26:165-177.
Warlcn, S.M. 1964. Some aspects of the life history of Cyprinodon variegatus
Lacep5del803, in southern Delaware. Masters Thesis. University of Delaware,
Newark, DE. 44 p.
Weber, R.E. 1981. Intraspccific adaptation of hemoglobin function in fish to oxygen
availability. Pages 87-102 in A.D.F. Addink and N. Spronk, editors. Exogenous
and endogenous influences on metabolic and neural control, Volume 1. Pergamon
Press. Oxford.
Weber, R.E. 1990. Functional significance and structural basis of multiple hemoglobins
with special reference to cctothermic vertebrates. Pages 58-75 in J.P Truchot and
B. Lahlau, editors. Animal nutrition and transport processes, Volume 2. Transport,
respiration and excretion: comparative and environmental aspects. Kargcr, Basel.
188 p.
Weber, R.E., G. Lykkeboe, and K. Johansen. 1976. Physiological properties of cel
haemoglobin: hypoxic acclimation, phosphate effects and multiplicity. Journal of
Experimental Biology 64:75-88.
Wells, N.A. 1935. The influence of temperature upon the respiratory metabolism of the
pacific killifish, Fundulus parvipinnis. Physiological Zoology 8:195-227.

122
Wells, R.M.G., M.D. Ashby, S.J. Duncan, and J.A. MacDonald. 1980. Comparative study
of the erythrocytes and haemoglobins in nototheniid fishes from Antarctica. Journal
of Fish Biology 17:517-527.
Wells, R.M.G., G.C. Grigg, L.A. Beard, and G. Summers. 1989. Hypoxic responses in a
fish from a stable environment: blood oxygen transport in the antarctic fish
Pagothenia borchgrevinki. Journal of Experimental Biology 141:97-111.
Wells, R.M.G., and R.E. Weber. 1991. Is there an optimal haematocrit for rainbow trout,
Oncorhynchus mykiss (Walbaum)? An interpretation of recent data based on blood
viscosity measurements. Journal of Fish Biology 38:53-65.
Wheatly, M.G. 1988. Integrated responses to salinity fluctuation. American Zoologist
28:65-77.
Wilde, G.R., and A.A. Echelle. 1992. Genetic status of Pecos pupfish populations after
establishment of a hybrid swarm involving an introduced congener. Transactions of
the American Fisheries Society 121:277-286.
Winberg, G.G. 1956. Rate of metabolism and food requirements of fishes. Translation
Series No. 194. Fisheries Research Board of Canada Biological Station. Nanaimo,
British Columbia.
Winer, B.J., D.R. Brown, and K.M. Michels. 1991. Statistical principles in experimental
design. McGraw-Hill. New York, NY. 1057 p.
Woo, N.Y.S., and R.S.S. Wu. 1982. Metabolic and osmoregulatory changes in response to
reduced salinities in the red grouper, Epinephalus akaara (Temminck and Schlegel),
and the black sea bream, Mylio macrocephalus (Basilewsky). Journal of
Experimental Marine Biology and Ecology 65:139-161.
Wood, C.M., and W.S. Marshall. 1994. Ion balance, acid-base regulation, and chloride cell
function in the common killifish, Fundulus heteroclitus—a common euryhaline
estuarine teleost. Estuaries 17:34-52.
Wright, R.A., L.B. Crowder, and T.H. Martin. 1993. The effects of predation on the
survival and size-distribution of estuarine fishes: an experimental approach.
Environmental Biology of Fishes 36:291-300.
Yeager, D.P., and G.R. Ultsch. 1989. Physiological regulation and conformation: A BASIC
program for the determination of critical points. Physiological Zoology 62:888-907.
Yoshikawa, J.S.M., S.D. McCormick, G. Young, and H.A. Bern. 1993. Effects of salinity
on chloride cells and Na+, K+ -ATPasc activity in the teleost Gillichthys mirabilis.
Comparative Biochemistry and Physiology 105A:311-317.
Zadunaisky, J. 1984. The chloride cell: the active transport of chloride and the paracellular
pathways. Pages 129-176 in W.S. Hoar and D.J. Randall, editors. Fish physiology,
Volume 10B. Academic Press, London. 416 p.

BIOGRAPHICAL SKETCH
Dennis Charles Haney was bom March 18, 1962, in Inglewood, California to
Charles and Jeanne Haney. He grew up in Southern California with his parents and older
brother Scott, graduating from Chatsworth High School in 1979. Dennis began his career in
the biological sciences as an undergraduate at the University of California, San Diego
(UCSD). While at UCSD Dennis had his first true exposure to the joys (and pitfalls) of
research and teaching. He graduated in 1983 with a B.A. in biology (specialization in animal
physiology), and moved on to the graduate program at Oregon State University (OSU).
Dennis spent two years at OSU, where he was introduced to the study of fish physiology.
For his master's thesis he examined the physiological and hematological effects of
erythrocytic necrosis virus on chum salmon (Oncorhynchus keta). Dennis completed his
M.S. in biological science in 1985, following which he moved across the country to begin a
doctoral program at the University of Florida (UF). Since 1990 Dennis has simultaneously
worked on his dissertation and for the Department of the Interior as a Biological
Technician. Dennis completed his dissertation and graduated from UF in 1995.
123

I certify that I have read this study and that in my opinion it conforms to acceptable
standards of scholarly presentation and is fully adequate, in scope and quality, as a
dissertation for the degree of Doctor of Philosophy.
Frank‘Ñofcllíe, Chair
Professor of Zoology
I certify that I have read this study and that in my opinion it conforms to acceptable
standards of scholarly presentation and is fully adequate, in scope and quality, as a
dissertation for the degree of Doctor of Philosophy.
\
(by Lili 'white'
â– Harvey Lili
Professor o
'white
Zoology
I certify that I have read this study and that in my opin on it conforms to acceptable
standards of scholarly presentation and is fully adequate, in scope and quality, as a
dissertation for the degree of Doctor of Philosophy.
Brían^MxíÍíab
Professor of Zoology
I certify that I have read this study and that in my opinion it conforms to acceptable
standards of scholarly presentation and is fully adequate, in scope and quality, as a
dissertation for the degree of Doctor of Philosophy.
StephenWalsh
Courtesy Assistant Professor of Zoology
I certify that I have read this study and that in my oj
standards of scholarly presentation and is fully adequate, y
dissertation for the degree of Doctor of Philosophy.
nion it conforms to acceptable
scope and quality, as a
jSi/L
inneth Sulak
íourtesy Associate Professor of
Fisheries and Aquatic Sciences

I certify that I have read this study and that in my opinion it conforms to acceptable
standards of scholarly presentation and is fully adequate, in scope and quality, as a
dissertation for the degree of Doctor of Philosophy.
Tnomas Crisman
Professor of Environmental Engineering
Sciences
This dissertation was submitted to the Graduate Faculty of the Department of
Zoology in the College of Liberal Arts and Sciences and to the Graduate School and was
accepted as partial fulfillment of the requirements for the degree of Doctor of Philosophy.
December, 1995
Dean, Graduate School

17QH
Ja / W
i S j5
UNIVERSITY OF FLORIDA
3 1262 08554 5498



hUtf
UNIVERSITY OF FLORIDA
3 1262 08554 5498



PAGE 1

$ e 3&UI&" 5(63216(6 0$'( %< 7+( 6$/7 0$56+ 7(/(267 &<3512'21 9$5,(*$786 $7+(5,120253+$ &<35,12'217,'$(f 72 /,)( ,1 $ 9$5,$%/( 6$/,1,7< (19,5210(17 %\ '(11,6 &+$5/(6 +$1(< $ ',66(57$7,21 35(6(17(' 72 7+( *5$'8$7( 6&+22/ 2)7+( 81,9(56,7< 2) )/25,'$ ,1 3$57,$/ )8/),//0(17 2) 7+( 5(48,5(0(176 )25 7+( '(*5(( 2) '2&725 2) 3+,/2623+< 81,9(56,7< 2) )/25,'$

PAGE 2

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f DQG /RX 6RPPD ZHUH DOO LQYDOXDEOH LQ KHOSLQJ PH WR PDLQWDLQ JRRG PHQWDO KHDOWK RYHU WKH \HDUV :H VSHQW PDQ\ KRXUV WU\LQJ WR ILJXUH RXW OLIH WKH XQLYHUVH DQG HYHU\WKLQJ :H DOVR KDG ORWV RI IXQ ORRNLQJ IRU XQVXVSHFWLQJ ILVK DQG KHUSV /LVD *UHJRU\ KDV EHHQ D IULHQG FRXOG DOZD\V FRXQW RQ QR PDWWHU ZKDW -RKQ %LQHOOR ZDV P\ WUXVWHG ILHOG FRPSDQLRQ :H VSHQW PDQ\ GD\V WUDPSLQJ WKURXJK WKH VDOW PDUVK DQG RWKHU ERGLHV RI

PAGE 3

ZDWHUf LQ VHDUFK RI QXPHURXV ILVKHV LQFOXGLQJ WKH ZLO\ SXSILVK VDLOILQ PROO\ )ORULGD IODJILVK DQG IDW VOHHSHU 7KDQNV DOVR JR WR /HR 1LFR $GHOH +HQVOH\ 3DP )XOOHU 3DXOD &XVKLQJ 3DWULFLD +DUULVRQ /LDQQD -DUHFNL %HFN\ 7KRPSVRQ 9LQFH 'H0DUFR .HYLQ %DOGZLQ (OOHQ %XUURXJKV &KULV .DUGLVK 0DUN +RVWHWOHU 'RXJ :HDYHU )UDQN -RUGDQ -RKQ $QGHUVRQ &DURO %LQHOOR DQG HYHU\RQH HOVH IURP =RRORJ\ )RU WKH SDVW \HDUV KDYH ZRUNHG ZLWK PDQ\ SHRSOH DW WKH 'HSDUWPHQW RI WKH ,QWHULRUnV *DLQHVYLOOH ODERUDWRU\ SUHVHQWO\ WKH 1DWLRQDO %LRORJLFDO 6HUYLFH IRUPHUO\ 86 )LVK DQG :LOGOLIH 6HUYLFHf :KLOH ZRUNLQJ IXOOWLPH IRU WKH SDVW IHZ \HDUV FHUWDLQO\ VORZHG GRZQ P\ SURJUHVV WRZDUGV ILQLVKLQJ P\ 3K' WKH H[WUD WLPH ZDV ZRUWK LW PRVWO\f IRUPHG PDQ\ QHZ IULHQGVKLSV DQG OHDUQHG ORWV RI WKLQJV ZRXOGQnW KDYH RWKHUZLVH -LP :LOOLDPV VKRZHG PH WKDW IUHVKZDWHU FODPV ZHUH DFWXDOO\ NLQG RI QHDW 1RHO %XUNKHDG KHOSHG UHPLQG PH WKDW ILVK ZHUH VWLOO PXFK FRROHU /HV 3DUNHU -D\QH %ULP%R[ DQG KDG JUHDW IXQ GLYLQJ LQ ]HUR YLVLELOLW\ ZDWHU +RZDUG -HONV *DU\ +LOO $QQ )RVWHU 7LQD
PAGE 4

7$%/( 2) &217(176 /,67 2) 7$%/(6 YL /,67 2) ),*85(6 Y $%675$&7 [ &+$37(56 ,1752'8&7,21 ([SHULPHQWDO $QLPDO 4XHVWLRQV 6WXG\ 6LWH ,1)/8(1&( 2) (19,5210(17$/ 6$/,1,7< 21 5287,1( 0(7$%2/,& 5$7( $1' &5,7,&$/ 2;<*(1 7(16,21 2) &<35,12'21 9$5,(*$786 ,QWURGXFWLRQ 0HWKRGV 5HVXOWV 'LVFXVVLRQ ,1)/8(1&( 2) 6,08/$7(' 7,'$/ &+$1*(6 ,1 $0%,(17 6$/,1,7< 21 5287,1( 0(7$%2/,& 5$7( ,1 &<35,12'21 9$5,(*$786 ,QWURGXFWLRQ 0HWKRGV 5HVXOWV 'LVFXVVLRQ ,1)/8(1&( 2) $ )/8&78$7,1* 6$/,1,7< 5(*,0( 21 26025(*8/$7,21 ,1 &<35,12'21 9$5,(*$786 ,QWURGXFWLRQ 0HWKRGV 5HVXOWV 'LVFXVVLRQ ,1)/8(1&( 2) (19,5210(17$/ 6$/,1,7< 21 %/22' 2;<*(1 /(9(/6 2) &<35,12'21 9$5,(*$786 ,QWURGXFWLRQ 0HWKRGV ,9

PAGE 5

5HVXOWV 'LVFXVVLRQ 6800$5< $1' &21&/86,216 $33(1',&(6 &5,7,&$/ 2;<*(1 7(16,21 ),*85(6 ),(/' 0($685(0(176 /,7(5$785( &,7(' %,2*5$3+,&$/ 6.(7&+ Y

PAGE 6

/,67 2) 7$%/(6 7DEOH SDJH 3K\ORJHQHWLF FODVVLILFDWLRQ RI WKH F\SULQRGRQWLIRUP ILVKHV PRGLILHG IURP 3DUHQWL f 5HODWLRQVKLSV RI URXWLQH PHWDEROLVP 505f FULWLFDO R[\JHQ WHQVLRQ 3Ff DQG VORSH LQ WKH FRQIRUPDWLRQ UHJLRQ DW D VHULHV RI DPELHQW VDOLQLWLHV 9DOXHV DUH JLYHQ DV PHDQV s VH 0HDVXUHPHQWV RI R[\JHQ FRQFHQWUDWLRQ PJ /fOf VDOLQLW\ SSWf DQG WHPSHUDWXUH r&f WDNHQ DW IRXU VLWHV LQ WKH &HGDU .H\ DUHD IURP -XQH WKURXJK -XQH VHH WH[W IRU GHWDLOV RQ ORFDWLRQ RI VLWHVf 9DOXHV DUH JLYHQ DV PHDQV VH LQ SDUHQWKHVHVf VDPSOH VL]H $FFOLPDWLRQ DQG ILQDO VDOLQLWLHV XVHG LQ VLPXODWHG WLGDO FKDQJH VWXG\ 0HDQ URXWLQH PHWDEROLVP PJ &E KnOf EHIRUH DFFOLPDWLRQ VDOLQLW\f DQG IROORZLQJ ILQDO VDOLQLW\f D VLPXODWHG WLGDO FKDQJH 9DOXHV DUH JLYHQ DV PHDQV s VH *URXSV H[KLELWLQJ D VLJQLILFDQW FKDQJH LQ PHWDEROLVP DUH LQGLFDWHG ZLWK DQ DVWHULVN 6DOLQLW\ WULDOV XVHG LQ F\FOLFDO VDOLQLW\ VWXG\ 7KH JURXS PDLQWDLQHG DW SSW ZDV VSOLW LQWR WKUHH JURXSV IROORZLQJ F\FOH GD\ f JURXSV &T & DQG 4 VHH WH[W IRU GHWDLOVf 5HVXOWV RI VDOLQLW\ IOXFWXDWLRQV H[SHULPHQW 9DOXHV LQ WKH WRS URZ RI HDFK FHOO UHSUHVHQW KHPDWRFULW PHDVXUHPHQWV b HU\WKURF\WHVf YDOXHV LQ ERWWRP URZ RI HDFK FHOO UHSUHVHQW SODVPD RVPRODOLW\ PHDVXUHPHQWV P2VP NJnOf 6DPSOH VL]HV DUH Q IRU HDFK FHOO $OO YDOXHV DUH H[SUHVVHG DV PHDQV s VH 6HH WH[W IRU H[SODQDWLRQ RI JURXS DEEUHYLDWLRQV +HPDWRFULW +FWf KHPRJORELQ FRQFHQWUDWLRQ >+E@f HU\WKURF\WH FRXQW 5%&f PHDQ FRUSXVFXODU KHPRJORELQ 0&+f PHDQ FRUSXVFXODU YROXPH 0&9f DQG PHDQ FRUSXVFXODU KHPRJORELQ FRQFHQWUDWLRQ 0&+&f DV D IXQFWLRQ RI VDOLQLW\ IRU &\SULQRGRQ YDULHJDWXV $OO YDOXHV DUH H[SUHVVHG DV PHDQV s VH 6HH WH[W IRU IXUWKHU GHWDLOV RQ EORRG LQGLFHV YL

PAGE 7

/,67 2) ),*85(6 )LJXUH SDJH 0HDQ DGMXVWHG URXWLQH PHWDEROLF UDWHV 505f RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YDULHJDWXV PHWDEROLF UDWHV ZHUH PDVVDGMXVWHG XVLQJ DQ DQDO\VLV RI FRYDULDQFH EDUV LQGLFDWH s VH QXPHULFDO YDOXHV DERYH WKH SRLQWV LQ WKH ILJXUH LQGLFDWH VDPSOH VL]HV DW HDFK VDOLQLW\f 5HODWLRQVKLS EHWZHHQ PHDQ DGMXVWHG URXWLQH PHWDEROLF UDWHV 505f DQG PHDQ SODVPD RVPRODOLW\ RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YDULHJDWXV PHWDEROLF UDWHV ZHUH PDVVDGMXVWHG XVLQJ DQ DQDO\VLV RI FRYDULDQFH EDUV LQGLFDWH s VH SODVPD RVPRODOLW\ GDWD IURP 1RUGOLH f 0HDQ FULWLFDO R[\JHQ WHQVLRQ 3Ff PHDVXUHPHQWV RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YDULHJDWXV EDUV LQGLFDWH s VH QXPHULFDO YDOXHV DERYH WKH SRLQWV LQ WKH ILJXUH LQGLFDWH VDPSOH VL]HV DW HDFK VDOLQLW\f 5HODWLRQVKLS EHWZHHQ PHDQ DGMXVWHG URXWLQH PHWDEROLF UDWHV 505f DQG FULWLFDO R[\JHQ WHQVLRQV 3Ff RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YDULHJDWXV PHWDEROLF UDWHV ZHUH PDVVDGMXVWHG XVLQJ DQ DQDO\VLV RI FRYDULDQFH EDUV LQGLFDWH s VHf 5HODWLRQVKLS EHWZHHQ PHDQ FULWLFDO R[\JHQ WHQVLRQV 3Ff DQG PHDQ SODVPD RVPRODOLW\ RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YDULHJDWXV EDUV LQGLFDWH s VH SODVPD RVPRODOLW\ GDWD IURP 1RUGOLH tf 5HODWLRQVKLS EHWZHHQ PHDQ DGMXVWHG URXWLQH PHWDEROLF UDWHV 505f DQG PHDQ SODVPD RVPRODOLW\ RYHU D UDQJH RI VDOLQLWLHV LQ $GLQLD [HQLFD PHWDEROLF UDWHV ZHUH PDVVDGMXVWHG XVLQJ DQ DQDO\VLV RI FRYDULDQFH EDUV LQGLFDWH s VH QXPHULFDO YDOXHV DERYH WKH SRLQWV LQ WKH ILJXUH LQGLFDWH VDPSOH VL]HV DW HDFK VDOLQLW\f 6FKHPDWLF GLDJUDP RI UHVSLURPHWU\ DSSDUDWXV XVHG IRU URXWLQH PHWDEROLVP H[SHULPHQWV 6HH WH[W IRU GHWDLOHG GHVFULSWLRQ RI V\VWHP 5HVXOWV RI PHWDEROLF WULDOV ZKHUH VDOLQLW\ ZDV LQFUHDVHG RYHU WKH FRXUVH RI WKH WULDO %DUV UHSUHVHQW JURXSV OLVWHG LQ 7DEOH IRU ZKLFK ILQDO VDOLQLW\ ZDV JUHDWHU WKDQ LQLWLDO VDOLQLW\ 7KH KHLJKW RI HDFK EDU VLJQLILHV WKH PDJQLWXGH RI WKH VDOLQLW\ FKDQJH IRU HDFK PHWDEROLF WULDO DQG WKH DVWHULVN LQGLFDWHV DW ZKLFK RI WKH VDOLQLWLHV IRU HDFK PHWDEROLF WULDOf WKH URXWLQH PHWDEROLF UDWH 505f ZDV KLJKHVW 7KH [ D[LV KDV QR VFDOH DQG VHUYHV RQO\ WR YLVXDOO\ VHSDUDWH JURXSV YLL

PAGE 8

5HVXOWV RI PHWDEROLF WULDOV ZKHUH VDOLQLW\ ZDV GHFUHDVHG RYHU WKH FRXUVH RI WKH WULDO %DUV UHSUHVHQW JURXSV OLVWHG LQ 7DEOH IRU ZKLFK ILQDO VDOLQLW\ ZDV OHVV WKDQ LQLWLDO VDOLQLW\ 7KH KHLJKW RI HDFK EDU VLJQLILHV WKH PDJQLWXGH RI WKH VDOLQLW\ FKDQJH IRU HDFK PHWDEROLF WULDO DQG WKH DVWHULVN LQGLFDWHV DW ZKLFK RI WKH VDOLQLWLHV IRU HDFK PHWDEROLF WULDOf WKH URXWLQH PHWDEROLF UDWH 505f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f FRXQW RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YDULHJDWXV EDUV LQGLFDWH s VH QXPHULFDO YDOXHV DERYH WKH SRLQWV LQ WKH ILJXUH LQGLFDWH VDPSOH VL]HV DW HDFK VDOLQLW\f 0HDQ KHPRJORELQ FRQFHQWUDWLRQ >+E@f RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YDULHJDWXV EDUV LQGLFDWH s VH QXPHULFDO YDOXHV DERYH WKH SRLQWV LQ WKH ILJXUH LQGLFDWH VDPSOH VL]HV DW HDFK VDOLQLW\f 0HDQ KHPDWRFULW +FWf RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YDULHJDWXV EDUV LQGLFDWH s VH QXPHULFDO YDOXHV DERYH WKH SRLQWV LQ WKH ILJXUH LQGLFDWH VDPSOH VL]HV DW HDFK VDOLQLW\f $ 3ORW LQGLFDWLQJ WKH FDOFXODWLRQ RI WKH FULWLFDO R[\JHQ WHQVLRQ 3Ff IRU DQ LQGLYLGXDO &\SULQRGRQ YDULHJDWXV LQ ZDWHU DW SSW $ 3ORW LQGLFDWLQJ WKH FDOFXODWLRQ RI WKH FULWLFDO R[\JHQ WHQVLRQ 3Ff IRU DQ LQGLYLGXDO &\SULQRGRQ YDULHJDWXV LQ ZDWHU DW SSW $ 3ORW LQGLFDWLQJ WKH FDOFXODWLRQ RI WKH FULWLFDO R[\JHQ WHQVLRQ 3Ff IRU DQ LQGLYLGXDO &\SULQRGRQ YDULHJDWXV LQ ZDWHU DW SSW $ *HQHUDOL]HG FULWLFDO R[\JHQ WHQVLRQ 3Ff SORWV DW HDFK VDOLQLW\ XVHG LQ WKH 3F H[SHULPHQWV 3ORWV ZHUH SURGXFHG E\ XVLQJ WKH PHDQ 3F PHDQ URXWLQH PHWDEROLF UDWH 505f DQG PHDQ VORSH LQ WKH FRQIRUPDWLRQ UHJLRQ IRU HDFK VDOLQLW\ JURXS YLLL

PAGE 9

$ 2[\JHQ FRQFHQWUDWLRQ PJ /Of VDOLQLW\ SSWf DQG WHPSHUDWXUH r&f DW IRXU VLWHV LQ WKH &HGDU .H\ DUHD WDNHQ EHWZHHQ -XQH DQG -XQH 9DOXHV DUH JLYHQ DV PHDQV EDUV LQGLFDWH s VH Df 0HDVXUHPHQWV WDNHQ RQ WKH ERWWRP DW VLWH Ef PHDVXUHPHQWV WDNHQ RQ WKH VXUIDFH DW VLWH Ff 0HDVXUHPHQWV WDNHQ RQ WKH ERWWRP DW VLWH Gf PHDVXUHPHQWV WDNHQ RQ WKH VXUIDFH DW VLWH Hf 0HDVXUHPHQWV WDNHQ RQ WKH ERWWRP DW VLWH PHDVXUHPHQWV WDNHQ RQ WKH VXUIDFH DW VLWH Jf 0HDVXUHPHQWV WDNHQ RQ WKH VXUIDFH DW VLWH ,;

PAGE 10

$EVWUDFW RI 'LVVHUWDWLRQ 3UHVHQWHG WR WKH *UDGXDWH 6FKRRO RI WKH 8QLYHUVLW\ RI )ORULGD LQ 3DUWLDO )XOILOOPHQW RI WKH 5HTXLUHPHQWV IRU WKH 'HJUHH RI 'RFWRU RI 3KLORVRSK\ 5(63216(6 0$'( %< 7+( 6$/7 0$56+ 7(/(267 &<35,12'21 9$5,(*$786 $7+(5,120253+$ &<3512'217'$(f 72 /,)( ,1 $ 9$5,$%/( 6$/,1,7< (19,5210(17 %\ '(11,6 &+$5/(6 +$1(< 'HFHPEHU &KDLUPDQ )UDQN 1RUGOLH 0DMRU 'HSDUWPHQW =RRORJ\ &\SULQRGRQ YDULHJDWXV D FRPPRQ FRDVWDO UHVLGHQW RI WKH ZHVWHUQ $WODQWLF 2FHDQ DQG *XOI RI 0H[LFR OLYHV LQ DPELHQW VDOLQLWLHV UDQJLQJ IURP IUHVKZDWHU WR SSW )LVK XVHG LQ WKLV VWXG\ ZHUH REWDLQHG IURP D *XOI RI 0H[LFR VDOW PDUVK QHDU &HGDU .H\ )ORULGD ,Q D VWHDG\VWDWH H[SHULPHQW URXWLQH PHWDEROLF UDWH 505f DQG FULWLFDO R[\JHQ WHQVLRQ 3Ff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

PAGE 11

VDOLQLW\ ,QGLYLGXDOV RI & YDULHJDWXV UHVSRQGHG WR IOX[HV DW VDOLQLW\ H[WUHPHV E\ UHGXFLQJ JHQHUDO DFWLYLW\ DQG HQHUJ\ H[SHQGLWXUHVf§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f DQG ORZHVW SSWf VDOLQLWLHV WHVWHG 2[\JHQ FDUU\LQJ FDSDFLW\ ZDV KLJKHVW LQ WKH JURXS DFFOLPDWHG WR SSW (U\WKURF\WH FRXQW ZDV WKH PRVW FRQVLVWHQW LQGLFDWRU RI WKH LQIOXHQFH RI VDOLQLW\ RQ EORRG R[\JHQ ZLWK KHPDWRFULW WKH OHDVW FRQVLVWHQW PHDVXUH [L

PAGE 12

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f )DFWRUV WKDW PD\ LQIOXHQFH VDOLQLW\ LQ VDOW PDUVKHV LQFOXGH SUHFLSLWDWLRQ ZLQG IUHTXHQF\ DQG H[WHQW RI WLGHV VKRUHOLQH KHLJKW DQG FRDVWDO WRSRJUDSK\ 5HPDQH DQG 6FKOLHSHU :KHDWO\ f 'XH WR WKH KDUVKQHVV RI WKH SK\VLFDO HQYLURQPHQW VDOW PDUVKHV WHQG WR KDYH ORZ VSHFLHV GLYHUVLW\ EXW RIWHQ KLJK DEXQGDQFH RI VHOHFWHG VSHFLHV 0F&,XVN\ 'XQVRQ DQG 7UDYLV f :KHUHDV YDULDEOH VDOLQLW\ KDELWDWV DUH DOVR FKDUDFWHUL]HG E\ IOXFWXDWLRQV LQ RWKHU HQYLURQPHQWDO IDFWRUV VXFK DV WHPSHUDWXUH GLVVROYHG R[\JHQ DQG S+ 9HPEHUJ :KHDWO\ f WKH GLVWULEXWLRQ DQG DEXQGDQFH RI ILVKHV LQ WKHVH KDELWDWV LV ODUJHO\ GHWHUPLQHG E\ VDOLQLW\ 0F&,XVN\ 'DYHQSRUW DQG 6D\HU *LOO DQG 3RWWHU f 1RW VXUSULVLQJO\ DQLPDOV UHVSRQG WR IOXFWXDWLRQV LQ VDOLQLW\ LQ FRPSOH[ ZD\V 6DOLQLW\ DIIHFWV RVPRUHJXODWLRQ YHQWLODWLRQ PHWDEROLVP DFLGEDVH EDODQFH JURZWK UHSURGXFWLRQ GHYHORSPHQW DQG RWKHU ELRORJLFDO SURFHVVHV :KHDWO\ f 6RPH RI WKH SULPDU\ UHVSRQVHV RI WHOFRVWV WR FKDQJHV LQ VDOLQLW\ DUH UHYLHZHG KHUH

PAGE 13

$YRLGDQFH LV WKH ILUVW OLQH RI GHIHQVH WR YDULDEOH VDOLQLW\ FRQGLWLRQV ,I EHKDYLRUDO UHVSRQVHV GR QRW VXIILFLHQWO\ PLQLPL]H H[SRVXUH WR YDULDEOH VDOLQLWLHV DTXDWLF RUJDQLVPV PXVW UHO\ RQ SK\VLRORJLFDO DQG ELRFKHPLFDO UHVSRQVHV WR WROHUDWH HQYLURQPHQWDO FKDQJHV %HLWLQJHU DQG 0F&DXOH\ f 7KLV PD\ EH E\ SDVVLYH WROHUDQFH RVPRFRQIRUPLQJf RU DFWLYH RVPRUHJXODWLRQ 7UXFKRW 0F&OXVN\ f 2QO\ ILVKHV WKDW DUH FDSDEOH RI RVPRUHJXODWLRQ FDQ WROHUDWH ZLGH FKDQJHV LQ VDOLQLW\ DQG WKHVH HXU\KDOLQH WHOHRVWV ZLOO EH WKH IRFXV RI WKH UHVW RI WKLV UHYLHZ 7KH EDVLF SDWWHUQV RI RVPRUHJXODWLRQ LQ ILVKHV KDYH EHHQ H[WHQVLYHO\ UHYLHZHG LQ UHFHQW \HDUV 3DUYDWKHVZDUDUDR (GG\ (YDQV =DGXQDLVN\ .DPDN\ )RVNHWW 3LVDP DQG 5DPERXUJ 9HQWUHOOD HW DO (YDQV 0F&RUPLFN :RRG DQG 0DUVKDOO f (XU\KDOLQH WHOHRVWV UHJXODWH WKHLU EORRG RVPRODOLW\ DW DERXW RQHWKLUG WKH FRQFHQWUDWLRQ RI VHDZDWHU SSWf DQG WKXV IDFH VHYHUH RVPRWLF SUREOHPV ZKHWKHU LQ IUHVKZDWHU SSWf RU VHDZDWHU %RG\ IOXLGV RI D WHOHRVW LQ IUHVKZDWHU DUH K\SHURVPRWLF WR WKH H[WHUQDO HQYLURQPHQW ZKHUHDV LQ VHDZDWHU WKH\ DUH K\SRRVPRWLF 7KXV HXU\KDOLQH ILVK SRVVHVV PHFKDQLVPV IRU RVPRUHJXODWLQJ LQ ERWK K\SHURVPRWLF DQG K\SRRVPRWLF FRQGLWLRQV 7HOHRVWV LQ VHDZDWHU DUH VXVFHSWLEOH WR D ORVV RI ERG\ ZDWHU WR WKH H[WHUQDO HQYLURQPHQW DQG EDODQFH ZDWHU ORVV E\ DFWLYHO\ GULQNLQJ ODUJH DPRXQWV RI VHDZDWHU +RZHYHU ERWK ZDWHU DQG VDOWV DUH DEVRUEHG WRJHWKHU DFURVV WKH JXW ,QJHVWHG H[FHVV VDOWV DUH DFWLYHO\ H[FUHWHG GLYDOHQW LRQV PRVWO\ LQ XULQH DQG IHFHV DQG PRQRYDOHQW LRQV E\ WKH JLOOV $FWLYH H[FUHWLRQ RI VDOWV E\ WKH JLOOV WDNHV SODFH YLD FKORULGH FHOOV =DGXQDLVN\ .DPDN\ )RVNHWW 3LVDP DQG 5DPERXUJ f &KORULGH FHOOV LQ VFDZDWFU DFFOLPDWFG ILVK DUH ORFDWHG DW WKH EDVH RI WKH VHFRQGDU\ JLOO ODPHOODH DQG DUH ODUJH FROXPQDUVKDSHG FHOOV XVXDOO\ H[WHQGLQJ IURP WKH EDVDO HSLWKHOLXP WR WKH H[WHUQDO HQYLURQPHQW 7KH\ DUF FKDUDFWHUL]HG E\ QXPHURXV PLWRFKRQGULD IRU WKLV UHDVRQ WKH\ DUF RIWHQ UHIHUUHG WR DV PLWRFKRQGULD ULFK FHOOVf DQG DQ H[WHQVLYH WXEXODU UHWLFXOXP FRQWLQXRXV

PAGE 14

ZLWK WKH EDVRODWHUDO PHPEUDQH 3LVDP DQG 5DPERXUJ f 7KH WUDQVSRUW HQ]\PH 1D.$73DVH LV UHVWULFWHG WR WKLV WXEXODU V\VWHP DQG WR WKH EDVRODWHUDO PHPEUDQH 7KLV LRQ SXPS FUHDWHV D ODUJH 1D JUDGLHQW ORZ LQ WKH FKORULGH FHOO F\WRSODVPf ZKLFK GULYHV D 1D&O FRWUDQVSRUWHU E\ ZKLFK &On HQWHUV WKH FHOO 7KH &O DFFXPXODWHV VXIILFLHQWO\ LQ WKH FHOO VXFK WKDW LW LV DEOH WR H[LW WR WKH H[WHUQDO HQYLURQPHQW DFURVV WKH DSLFDO PHPEUDQH ZLWK 1D IROORZLQJ SDVVLYHO\ GRZQ LWV HOHFWURFKHPLFDO JUDGLHQW EHWZHHQ DGMDFHQW FKORULGH FHOOV (YDQV 0F&RUPLFN :RRG DQG 0DUVKDOO f ,Q IUHVKZDWHU WHOHRVWV FRQWLQXRXVO\ IDFH DQ HIIOX[ RI VDOWV DQG DQ LQIOX[ RI ZDWHU +HQFH WKHLU RVPRUHJXODWRU\ UHVSRQVH LV WR DFWLYHO\ WUDQVSRUW VDOWV IURP WKH H[WHUQDO HQYLURQPHQW YLD WKH JLOOV WR DYRLG GULQNLQJ ZDWHU DQG WR H[FUHWH FRSLRXV DPRXQWV RI GLOXWH XULQH 2XU SUHVHQW XQGHUVWDQGLQJ RI WKH PHFKDQLVPV LQYROYHG LQ WKH XSWDNH RI 1D DQG &On IURP WKH HQYLURQPHQW LQ IUHVKZDWHU WHOHRVWV LV VRPHZKDW XQFOHDU DQG LQFRPSOHWH ,W DSSHDUV WKDW &O LV DFWLYHO\ WDNHQ XS ZLWK 1D XSWDNH EHLQJ D SDVVLYH SURFHVV :KHWKHU WKLV WDNHV SODFH YLD D IUHVKZDWHUW\SH FKORULGH FHOO RU WKURXJK LQWHUDFWLRQV ZLWK DFLGEDVH EDODQFH LV XQFOHDU DV HYLGHQFH H[LVWV IRU ERWK :RRG DQG 0DUVKDOO f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f 7KH KRUPRQH PRVW LQYROYHG ZLWK RVPRUHJXODWLRQ XQGHU IUHVKZDWHU FRQGLWLRQV LV SURODFWLQ ZLWK DUJLQLQH YDVRWRFLQ XURWHQVLQ ,, FDWHFKRODPLQHV DQG DWULDO

PAGE 15

QDWULXUHWLF SHSWLGH DOVR SOD\LQJ UROHV +LUDQR HW DO %HUQ DQG 0DGVHQ 7DNHL f &RQWURO RI RVPRUHJXODWLRQ LQ VHDZDWHU LV ODUJHO\ YLD WKH HIIHFWV RI FRUWLVRO DQG JURZWK KRUPRQH WRJHWKHU ZLWK WK\URLG KRUPRQHV DQJLRWHQVLQ ,, YDVRDFWLYH LQWHVWLQDO SHSWLGH DWULDO QDWULXUHWLF SHSWLGH DQG XURWHQVLQ %DOPHQW HWDO %HUQ DQG 0DGVHQ 7DNHL f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f 2QH UHDVRQ IRU D ODFN RI DJUHHPHQW DPRQJ VWXGLHV LV WKDW PDQ\ KDYH IRFXVHG RQ WKH HQHUJHWLF FRVWV DVVRFLDWHG ZLWK RVPRUHJXODWLRQ ZKLOH LJQRULQJ RWKHU UHODWHG SK\VLRORJLFDO DQG EHKDYLRUDO SURFHVVHV )RU H[DPSOH PDQ\ VWXGLHV KDYH XVHG PHDVXUHG GLIIHUHQFHV LQ PHWDEROLVP DW GLIIHUHQW VDOLQLWLHV WR UHSUHVHQW GLIIHUHQFHV LQ RVPRUHJXODWRU\ FRVWV 7KH UDWLRQDOH LV WKDW RVPRUHJXODWLRQ LV DQ HQHUJHWLFDOO\ FRVWO\ HYHQW ZLWK WKH HQHUJHWLF FRVWV EHLQJ UHODWHG WR WKH RVPRWLF JUDGLHQW EHWZHHQ WKH ILVK DQG LWV HQYLURQPHQW HJ 0DGDQ 0RKDQ 5DR 0XLU DQG 1LLPL )XUVSDQ HW DO =DGXQDLVN\ f +RZHYHU WKLV LV SUREDEO\ DQ RYHUVLPSOLILFDWLRQ VLQFH RWKHU IDFWRUV VXFK DV DFWLYLW\ IRRG LQWDNH DQG SHUPHDELOLW\ FKDQJHV PD\ DOVR EH LQIOXHQFHG E\ VDOLQLW\ DQG LQ WXUQ DIIHFW PHWDEROLVP 6ZDQVRQ f )RU WKLV UHDVRQ PHWDEROLF PHDVXUHPHQWV UHSUHVHQW WKH RYHUDOO FRVWV DVVRFLDWHG ZLWK OLYLQJ LQ D SDUWLFXODU VDOLQLW\ HQYLURQPHQW ZLWK FRPSDULVRQV DPRQJ VDOLQLWLHV UHIOHFWLQJ WKHVH WRWDO FRVWV QRW VLPSO\ WKH FRVW RI RVPRUHJXODWLRQ

PAGE 16

6HYHUDO DWWHPSWV KDYH EHHQ PDGH WR FDWHJRUL]H WKH SDWWHUQV RI PHWDEROLF UHVSRQVHV RI WHOHRVWV WR DOWHUHG VDOLQLWLHV .LQQH 5HPDQH DQG 6FKOLHSHU 1RUGOLH 0RUJDQ DQG ,ZDPD f 2QH FRPPRQO\ REVHUYHG SDWWHUQ LV WKDW WKH ORZHU UDWHV RI PHWDEROLVP LQ UHVSRQVH WR VDOLQLW\ DUH DVVRFLDWHG ZLWK HQYLURQPHQWV IRU ZKLFK WKH VSHFLHV DQG OLIH VWDJHf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f 9DULDWLRQV LQ R[\JHQ FDQ GLFWDWH WKH GLVWULEXWLRQ RI VRPH VSHFLHV LQ DTXDWLF HFRV\VWHPV %RXWLOLHU f &KDQJLQJ HQYLURQPHQWDO VDOLQLW\ PD\ GLUHFWO\ LQIOXHQFH UHVSLUDWRU\ IXQFWLRQ E\ DIIHFWLQJ ERWK WKH R[\JHQ VROXELOLW\ LQ ZDWHU SXPSHG RYHU WKH JLOOV DQG WKH VROXELOLW\ RI JDVHV GLVVROYHG LQ SODVPD &KDQJHV LQ WKH LRQLF FRPSRVLWLRQ RI WKH ERG\ IOXLGV FRXOG DOVR LQWHUDFW ZLWK R[\JHQ WR LQIOXHQFH WROHUDQFH WR YDULDEOH VDOLQLW\ FRQGLWLRQV 7UXFKRW f ([SRVXUH RI ILVK WR UHGXFHG R[\JHQ WHQVLRQV LQLWLDWHV SK\VLRORJLFDO UHVSRQVHV PRVWO\ GLUHFWHG DW LQFUHDVLQJ WKH DPRXQW RI R[\JHQ DYDLODEOH WR WKH WLVVXHV %RXWLOLHU HW DO f 7KH WUDQVIHU RI R[\JHQ IURP WKH HQYLURQPHQW WR WKH WLVVXHV FDQ EH FKDUDFWHUL]HG DV D VHULHV RI SURFHVVHV *LOO YHQWLODWLRQ LV WKH ILUVW VWHS IROORZHG LQ WXUQ E\ EUDQFKLDO GLIIXVLRQ EORRG R[\JHQ WUDQVSRUW DQG GLIIXVLRQ LQWR WKH WLVVXHV WKH ODVW RI ZKLFK GXH WR D ODFN RI DYDLODEOH LQIRUPDWLRQ ZLOO QRW EH GLVFXVVHG IXUWKHUf 3HUU\ DQG 0F'RQDOG f *LOO YHQWLODWLRQ LV D IXQFWLRQ RI WKH IUHTXHQF\ DQG GHSWK RI EUHDWKLQJ DQG LV QRUPDOO\ LQFUHDVHG ZKHQ ORZ R[\JHQ WHQVLRQV DUH HQFRXQWHUHG :KLOH YHQWLODWLRQ GRFV QRW

PAGE 17

DSSHDU WR EH GLUHFWO\ OLPLWLQJ WR WKH XSWDNH RI R[\JHQ VXEVWDQWLDO FRVW LV LQYROYHG ZLWK LQFUHDVHG YHQWLODWRU\ SXPSLQJ ZKLFK XOWLPDWHO\ PHDQV WKDW DQ\ DGGLWLRQDO R[\JHQ DFTXLUHG LV XVHG WR IXHO WKH YHQWLODWRU\ DSSDUDWXV LWVHOI %RXWLOLHU HW DO 0F0DKRQ &DPHURQ 3HUU\ DQG 0F'RQDOG f 7ZR SURFHVVHV DUH ODUJHO\ DYDLODEOH WR LQFUHDVH EUDQFKLDO R[\JHQ GLIIXVLRQ LQFUHDVHV LQ IXQFWLRQDO JLOO VXUIDFH DUHD DQG LQFUHDVHV LQ WKH PHDQ ZDWHU WR EORRG R[\JHQ SDUWLDO SUHVVXUH JUDGLHQW 7KLV WUDGHRII LV SDUWLFXODUO\ LPSRUWDQW LQ UHJDUG WR VDOLQLW\ DV ILVK LQ ZDWHUV RI ORZ R[\JHQ WHQVLRQ PXVW EDODQFH WKH DGYDQWDJH RI PD[LPL]LQJ EUDQFKLDO R[\JHQ GLIIXVLRQ ZLWK D GLVDGYDQWDJH LQ RVPRUHJXODWLRQ GXH WR WKH DFFRPSDQ\LQJ LQFUHDVHV LQ LRQ DQG ZDWHU H[FKDQJH 3HUU\ DQG 0F'RQDOG f ,QFUHDVLQJ EORRG JDV WUDQVSRUW LV OLNHO\ WKH SULPDU\ URXWH XVHG E\ PRVW ILVK WR LQFUHDVH WKH DPRXQW RI R[\JHQ GHOLYHUHG WR WKH WLVVXHV 2[\JHQ WUDQVSRUW E\ WKH EORRG LQ WHOHRVWV GHSHQGV RQ WKH UHVSLUDWRU\ SLJPHQW KHPRJORELQ %ORRG R[\JHQ WUDQVSRUW LV QRUPDOO\ LQFUHDVHG E\ LQFUHDVLQJ WKH FRQFHQWUDWLRQ RI KHPRJORELQ LQFUHDVLQJ WKH QXPEHU RI HU\WKURF\WHV LQ FLUFXODWLRQ DQGRU E\ DGMXVWLQJ WKH DIILQLW\ RI KHPRJORELQ IRU R[\JHQ 'DYLV :HOOV HW DO -HQVHQ HW DO 3HUU\ DQG 0F'RQDOG f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f %HORZ WKH 3F PHWDEROLVP LV GHSHQGHQW XSRQ R[\JHQ WHQVLRQ DQG GHFUHDVHV OLQHDUO\ ZLWK GHFUHDVHV LQ R[\JHQ 7KH 3F ZDV SUREDEO\ ILUVW GRFXPHQWHG E\ +DOO f EXW ZDV

PAGE 18

QRW FRQVLGHUHG DQ LPSRUWDQW LQGH[ IRU ILVKHV XQWLO IRUPDOL]HG E\ )U\ f +RZHYHU WKLV LV DQ H[WUHPHO\ LPSRUWDQW YDULDEOH UHODWLQJ WR KDELWDW VHOHFWLRQ DQG RYHUDOO PHWDEROLF SDWWHUQV RI ILVKHV DQG FDOFXODWLRQV RI WKH 3F KDYH VLQFH EHHQ PDGH IRU D QXPEHU RI VSHFLHV XQGHU D YDULHW\ RI HQYLURQPHQWDO FRQGLWLRQV ([SHULPHQWDO $QLPDO 7KH VXEMHFW RI WKLV VWXG\ ZDV WKH VKHHSVKHDG PLQQRZ &\SULQRGRQ YDULHJDWXV &\SULQRGRQ YDULHJDWXV LV D PHPEHU RI WKH IDPLO\ &\SULQRGRQWLGDH D ODUJH DQG GLYHUVH IDPLO\ FRQWDLQLQJ RYHU VSHFLHV LQ JHQHUD 3DUHQWL 3DUNHU DQG .RPILHOG VHH 7DEOH f 0HPEHUV RI WKLV IDPLO\ DUH IRXQG LQ IUHVK EUDFNLVK DQG VDOW ZDWHU DQG GLVWULEXWHG SDQWURSLFDOO\ DV ZHOO DV WKURXJKRXW 1RUWK $PHULFD &\SULQRGRQ YDULHJDWXV LV WKH W\SH VSHFLHV IRU ERWK WKH JHQXV &\SULQRGRQ DQG WKH IDPLO\ &\SULQRGRQWLGDH 7KH JHQXV &\SULQRGRQ FRPSULVHV D JURXS RI DSSUR[LPDWHO\ VSHFLHV RI VPDOO RYLSDURXV ILVKHV FRPPRQO\ NQRZQ DV SXSILVKHV 7KH\ H[KLELW UHPDUNDEOH WROHUDQFH WR KDUVK HQYLURQPHQWDO FRQGLWLRQV 0LOOHU f RFFXUULQJ WKURXJKRXW 1RUWK DQG &HQWUDO $PHULFD WKH &DULEEHDQ 6HD DQG 9HQH]XHOD 'DUOLQJ 7XUQHU DQG /LX 3DUHQWL 'XJJLQV HW DO %DUQV DQG :RKOJHPXWK f 7KH JHQXV LV FKDUDFWHUL]HG E\ OLPLWHG JHQHWLF GLYHUJHQFH DQG IHZ XQLTXH DOOHOHV HYHQ LQ PRUSKRORJLFDOO\ GLVWLQFW VSHFLHV .RGULF%URZQ f 0XFK RI WKH UHVHDUFK RQ WKH JHQXV KDV IRFXVHG RQ VSHFLHV LQKDELWLQJ 1RUWK $PHULFDQ GHVHUWV ZKHUH WKH ODUJHVW FRQFHQWUDWLRQ RI VSHFLHV RFFXUV 0RVW RI WKHVH GHVHUW VSHFLHV KDYH VPDOO DOORSDWULF GLVWULEXWLRQV 0LOOHU 'XJJLQV HW DO f 0HPEHUV RI WKLV JHQXV DOVR RFFXU LQ WKH FRDVWDO EUDFNLVK DQG PDULQH ZDWHUV RI HDVWHUQ 1RUWK $PHULFD ZKHUH & YDULHJDWXV LV WKH GRPLQDQW SXSILVK VSHFLHV &\SULQRGRQ YDULHJDWXV LV WKH RQO\ SXSILVK VSHFLHV KDYLQJ DQ H[WHQVLYH JHRJUDSKLF UDQJH ,W LV IRXQG DORQJ WKH $WODQWLF FRDVW IURP 0DVVDFKXVHWWV WR WKH )ORULGD .H\V DQG WKURXJKRXW WKH *XOI RI 0H[LFR $ GLVMXQFW SRSXODWLRQ RFFXUV DORQJ WKH
PAGE 19

7DEOH 3K\ORJHQHWLF FODVVLILFDWLRQ RI WKH F\SULQRGRQWLIRUP ILVKHV PRGLILHG IURP 3DUHQWL f 2UGHU &\SULQRGRQWLIRUPHV 6XERUGHU $SORFKHLORLGHL 6XERUGHU &\SULQRGRQWRLGHL 6HFWLRQ )DPLO\ 3URIXQGXOLGDH 6HFWLRQ 'LYLVLRQ )DPLO\ )XQGXOLGDH 'LYLVLRQ 6XSHUIDPLO\ 3RHFLOLRLGHD )DPLO\ $QDEOHSLGDH )DPLO\ 3RHFLOLLGDH 6XSHUIDPLO\ &\SULQRGRQWRLGFD )DPLO\ *RRGHLGDH )DPLO\ &\SULQRGRQWLGDH 6XEIDPLO\ &\SULQRGRQWLQDH 7ULEH 2UHVWLLQL *HQXV 2UHVWLDV *HQXV .RVVZLJLFKWK\V *HQXV $SKDQLXV 7ULEH &\SULQRGRQWLQL *HQXV &\SULQRGRQ *HQXV 0HJXSVLORQ *HQXV -RUGDQHOOD *HQXV )ORULGLFKWK\V *HQXV &XDODF

PAGE 20

%DKDPDV :HVW ,QGLHV DQG &XED 'XJJLQV HW DO f &\SULQRGRQ YDULHJDWXV KDV DOVR EHHQ LQWURGXFHG LQWR VHYHUDO DUHDV ZKHUH WKH\ KDYH QHJDWLYHO\ LPSDFWHG QDWLYH SXSILVK VSHFLHV (FKHOOH DQG (FKHOOH (FKHOOH DQG &RQQRU .RGULF%URZQ :LOGH DQG (FKHOOH f *HQHWLF YDULDELOLW\ ZLWKLQ & YDULHJDWXV LV DV ODUJH DV WKH DPRXQW RI JHQHWLF GLYHUJHQFH ZLWKLQ WKH HQWLUH JHQXV &\SULQRGRQ ZLWK PRVW RI WKLV JHQHWLF GLYHUJHQFH RFFXUULQJ LQ SRSXODWLRQV QRUWK RI &DSH +DWWHUDVf§YHU\ OLWWOH JHQHWLF GLYHUJHQFH LV GLVSOD\HG DPRQJ VRXWKHUQ SRSXODWLRQV 'DUOLQJ 6FKZDUW] HW DO f &\SULQRGRQ YDULHJDWXV LV D QXPHULFDOO\ GRPLQDQW DQG HFRORJLFDOO\ LPSRUWDQW VSHFLHV WKURXJKRXW PRVW RI LWV UDQJH HVSHFLDOO\ LQ VDOW PDUVK DQG HVWXDULQH ZDWHUV .LOE\ 6LPSVRQ DQG *XQWHU 5HO\HD 1DXJKWRQ DQG 6DORPDQ 6XEUDKPDQ\DP DQG &RXOWDV 6WRXW 1HOVRQ 5RVV DQG 'RKHUW\ f 'XH WR LWV LPSRUWDQFH & YDULHJDWXV KDV EHHQ DQ LPSRUWDQW UHVHDUFK DQLPDO LQ GLYHUVH GLVFLSOLQHV 7KHVH LQFOXGH LQYHVWLJDWLRQV RQ WKH VSHFLHV EHKDYLRU ,W]NRZLW] ,W]NRZLW] 0HWWHH DQG %HFNKDP ,W]NRZLW] 'Z\HU DQG %FXOLJ f HFRORJ\ 'ROO DQG %DVW 0DUWLQ 0DUWLQ $EOH +DUULQJWRQ DQG +DUULQJWRQ )\IH 6KLSOH\ $YLOD HWDO :ULJKW HW DO f HYROXWLRQ (OGHU DQG 7XUQHU f OLIH KLVWRU\ :DUOFQ 'F 9ODPLQJ HW DO %HUU\ $EOH (FKHOOH DQG (FKHOOH f SK\VLRORJ\ 0DUWLQ .DUQDN\ HW DO 6XEUDKPDQ\DP 1RUGOLH %DUWRQ DQG %DUWRQ 1RUGOLH 3HWHUVRQ DQG *LOPRUH 1RUGOLH DQG :DOVK 3HWHUVRQ 3QFH HW DO 1RUGOLH HW DO 'XQVRQ HW DO f DQG UHSURGXFWLRQ 5DQH\ HWDO :DUOHQ %HUU\ .RGULF%URZQ &RQRYHU DQG 'F0RQG f 7KH VSHFLHV KDV DOVR EHHQ XVHG LQ YROXPLQRXV WR[LFRORJ\ H[SHULPHQWV EHFDXVH RI LWV H[WUHPH KDUGLQHVV HJ 6FKLPQLHO DQG +DQVHQ +DZNLQV HW DO %DWWDORUD HW DO /LQWRQ f &\SULQRGRQ YDULHJDWXV KDV EHHQ FDOOHG WKH WRXJKHVW ILVK LQ 1RUWK $PHULFD *XQWHU f GXH WR LWV H[WUHPH WROHUDQFH RI KDUVK HQYLURQPHQWDO FRQGLWLRQV ,W LV IRXQG LQ ZDWHUV UDQJLQJ IURP IUHVKZDWHU $JHU -RKQVRQ .XVKODQ f WR VDOLQLWLHV RI

PAGE 21

SSW 6LPSVRQ DQG *XQWHU f DOWKRXJK LW W\SLFDOO\ LQKDELWV EUDFNLVK ZDWHU DQG FRDVWDO VDOW PDUVKHV &\SULQRGRQ YDULHJDWXV LV WROHUDQW RI WHPSHUDWXUHV UDQJLQJ IURP DERXW r& %HUU\ f WR r& 6WUDZQ DQG 'XQQ f DQG R[\JHQ OHYHOV DSSURDFKLQJ DQR[LD 2GXP DQG &DOGZHOO f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f ZLOO XVH WKH WHUP DGDSWDWLRQ LQ LWV EURDGHVW VHQVH GHILQLQJ LW DV D PRGLILFDWLRQ RI WKH FKDUDFWHULVWLFV RI DQ RUJDQLVP WKDW IDFLOLWDWH DQ HQKDQFHG DELOLW\ WR VXUYLYH DQG UHSURGXFH LQ D SDUWLFXODU HQYLURQPHQW ,Q WKLV ZD\ UHFRJQL]H WKDW DGDSWDWLRQV LQYROYH ERWK JHQHWLF DQG SK\VLRORJLFDO SKHQRW\SLFf FRPSRQHQWV ZKLOH QRW DWWHPSWLQJ WR VHSDUDWH WKHVH FRPSRQHQWV IURP RQH DQRWKHU 7KH WHUP DFFOLPDWLRQ ZLOO EH XVHG DV GHILQHG E\ 3URVVHU f ZKHUH FRPSHQVDWRU\ FKDQJHV DUH PHDVXUHG IROORZLQJ FKDQJHV LQ VLQJOH HQYLURQPHQWDO YDULDEOHV 4XHVWLRQV 7KLV VWXG\ ZDV GHVLJQHG WR H[DPLQH VRPH RI WKH FRVWV WR & YDULHJDWXV DVVRFLDWHG ZLWK OLYLQJ LQ YDULDEOH VDOLQLW\ HQYLURQPHQWV 6SHFLILFDOO\ DVNHG WKH IROORZLQJ TXHVWLRQV f :KDW DUH WKH PHWDEROLF FRVWV DVVRFLDWHG ZLWK GLIIHUHQW DPELHQW VDOLQLWLHV" f +RZ GRHV VDOLQLW\ LQIOXHQFH WKH HQHUJHWLF UHVSRQVH DW ORZ R[\JHQ WHQVLRQV" f :KDW LV WKH RVPRUHJXODWRU\ UHVSRQVH WR YDULDEOH VDOLQLW\ HQYLURQPHQWV" f +RZ GRFV VDOLQLW\ LQIOXHQFH EORRG R[\JHQ OHYHOV" 7KHVH TXHVWLRQV ZHUH WHVWHG E\ WKH IROORZLQJ PHDVXUHPHQW RI PHWDEROLVP LQ & YDULHJDWXV IXOO\ DFFOLPDWHG WR D ZLGH UDQJH RI H[SHULPHQWDO VDOLQLWLHV PHDVXUHPHQW RI WKH FULWLFDO R[\JHQ WHQVLRQ LQ & YDULHJDWXV IXOO\ DFFOLPDWHG WR WKH VDPH

PAGE 22

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f 7KLV UHVXOWV LQ D ZLGH LQWHUWLGDO ]RQH DORQJ WKH FRDVW )XUWKHUPRUH WKH FRDVWDO SK\VLRJUDSK\ LV H[WUHPHO\ GLYHUVH GXH LQ ODUJH SDUW WR LUUHJXODULWLHV LQ WKH VKRUH OLQH RI WKH PDLQODQG WR WKH SUHVHQFH RI QXPHURXV LVODQGV DQG R\VWHU EDUV LQ WKH WLGDO DUHD DQG WR WKH PD]H RI LQWHUWLGDO DQG VXEWLGDO FUHHNV DQG FKDQQHOV .LOE\ f 1R VLJQLILFDQW VHGLPHQW VRXUFHV DUH IRXQG LQ WKLV DUHD DQG WLGHV RFFXU RQ D VHPLGLXUQDO EDVLV 7KH GRPLQDQW HPHUJHQW YHJHWDWLRQ LQ WKH DUHD LV 6SDUWLQD DOWHUQLIORUD ZLWK WKH VDOW PDUVKHV GRPLQDWHG E\ -XQFXV URHPHULDQXV )LVK FRPPXQLWLHV RI WKH -XQFXV PDUVK DUH GRPLQDWHG E\ DWKHULQLIRUPV ZLWK & YDULHJDWXV )XQGXOXV VLPLOLV DQG 3RHFLOLD ODWLSLQQD PDNLQJ XS b RI WKH FDWFK WKURXJKRXW PRVW RI WKH \HDU .LOE\ 6LPSVRQ DQG *XQWHU 6WRXW SHUV REVf

PAGE 23

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
PAGE 24

2QH JHQHUDO SDWWHUQ RI ILVKHV WKDW LQKDELW YDULDEOH VDOLQLW\ HQYLURQPHQWV LV D VWDEOH PHWDEROLF UDWH RYHU D UDQJH RI VDOLQLWLHV ZLWK WKH UDQJH RI VDOLQLWLHV PRVW FRPPRQO\ WHVWHG EHWZHHQ IUHVKZDWHU SSWf DQG VHDZDWHU SSWf 0RUJDQ DQG KYDPD f 3K\VLRORJLFDO VWDELOLW\ HQDEOHV VXFK ILVK WR KDYH D HXU\KDOLQH H[LVWHQFH WKDW LV XQIHWWHUHG E\ ODUJH PHWDEROLF FRVWV DVVRFLDWHG ZLWK DGMXVWPHQW WR VDOLQLW\ FKDQJH +RZHYHU WKHUH DUH UHODWLYHO\ IHZ VWXGLHV RI WKH PHWDEROLF UHVSRQVH RI HXU\KDOLQH ILVKHV RYHU DQ HYHQ ZLGHU UDQJH RI VDOLQLWLHV WKDW WKH\ HQFRXQWHU LQ WKHLU QDWXUDO KDELWDWV )LVK PHWDEROLVP LV DOVR VWURQJO\ LQIOXHQFHG E\ WKH SDUWLDO SUHVVXUH RI R[\JHQ 32f 0HWDEROLVP RI ILVKHV LV LQGHSHQGHQW RI 32 DV WKH\ PDLQWDLQ D FRQVWDQW PHWDEROLF UDWH RYHU D UDQJH RI 32 H[WHQGLQJ GRZQZDUG IURP KLJK DWPRVSKHULF OHYHOV WR VRPH ORZHU OHYHO GHILQHG DV WKH FULWLFDO R[\JHQ WHQVLRQ 3Ff %HORZ WKH 3F FRQIRUPDWLRQ UHJLRQf PHWDEROLVP GHSHQGV RQ R[\JHQ WHQVLRQ DQG GHFUHDVHV OLQHDUO\ ZLWK GHFUHDVHV LQ R[\JHQ )U\ f 2[\JHQ FRQVXPSWLRQ GHFOLQHV DW WKH 3F EHFDXVH WKH JDV H[FKDQJH V\VWHP FDQ QR ORQJHU VXSSO\ ERWK WKH H[WUD GHPDQGV RI WKH UHVSLUDWRU\ V\VWHP DQG WKH R[\JHQ GHPDQGV RI WKH WLVVXHV +XJKHV f 6LQFH 3F LV D XVHIXO PHWDEROLF SDUDPHWHU FDOFXODWLRQV RI WKH 3F KDYH EHHQ PDGH IRU D QXPEHU RI ILVKHV XQGHU D YDULHW\ RI HQYLURQPHQWDO FRQGLWLRQV HJ +DOO 7DQJ 8OWVFK HW DO 2WW HW DO 6XEUDKPDQ\DP 'RQQHOO\ DQG 7RUUHV 5DQWLQ HW DO 1RQQRWWH HW DO f +RZHYHU WKH LQIOXHQFH RI DPELHQW VDOLQLW\ RQ WKH 3F KDV QRW EHHQ VSHFLILFDOO\ VWXGLHG 7KLV LV VXUSULVLQJ VLQFH VDOLQLW\ DQG R[\JHQ DUH LPSRUWDQW DELRWLF IDFWRUV WKDW PD\ DFW V\QHUJLVWLFDOO\ LQ DIIHFWLQJ PHWDEROLVP 7KLV VWXG\ H[DPLQHG WKH LQIOXHQFH RI D ZLGH UDQJH RI HQYLURQPHQWDO VDOLQLWLHV RQ URXWLQH PHWDEROLF UDWH 505f DQG 3F LQ & YDULHJDWXV K\SRWKHVL]HG WKDW ERWK 505 DQG 3F ZRXOG EH XQDIIHFWHG E\ DOWHUDWLRQV LQ VDOLQLW\ RYHU WKH UDQJH RI VDOLQLWLHV FRPPRQO\ HQFRXQWHUHG LQ QDWXUDO KDELWDWV RI & YDULHJDWXV 6DOLQLWLHV RXWVLGH WKLV UDQJH EXW ZLWKLQ WKH UDQJH NQRZQ WR EH WROHUDWHG ZHUH SUHGLFWHG WR FDXVH LQFUHDVHV LQ ERWK SDUDPHWHUV

PAGE 25

0HWKRGV )LVK XVHG LQ WKLV VWXG\ ZHUH REWDLQHG E\ VHLQLQJ FDQDOV DQG GLWFKHV LQ WKH VDOW PDUVK QHDU &HGDU .H\ )ORULGD *XOI RI 0H[LFRf 6SHFLPHQV ZHUH WUDQVSRUWHG EDFN WR WKH ODERUDWRU\ LQ / FDUER\V FRQWDLQLQJ ZDWHU IURP WKH FROOHFWLRQ VLWH 8SRQ DUULYDO DW WKH ODERUDWRU\ LQGLYLGXDOV ZHUH KHOG RYHUQLJKW LQ WKLV ZDWHU ZLWK FRQVWDQW DHUDWLRQ 7KH IROORZLQJ GD\ ILVK ZHUH WUDQVIHUUHG LQWR KROGLQJ WDQNV WR / DTXDULDf PDLQWDLQHG DW WKH VDOLQLW\ DW ZKLFK ILVK ZHUH FDSWXUHG DQG WUHDWHG SURSK\ODFWLFDOO\ IRU GD\V LQ D PJ /nO VROXWLRQ RI $FULILDYLQHp )ROORZLQJ WUHDWPHQW JURXSV RI DSSUR[LPDWHO\ ILVK ZHUH SODFHG LQWR H[SHULPHQWDO / DTXDULDf WDQNV FRQWDLQLQJ ZDWHU DW D VDOLQLW\ ZLWKLQ SSW RI WKDW LQ ZKLFK WKH\ ZHUH FROOHFWHG %RWK KROGLQJ DQG H[SHULPHQWDO WDQNV ZHUH HTXLSSHG ZLWK XQGHUJUDYHO ILOWUDWLRQ DQG FRQVWDQW DHUDWLRQ DQG ZHUH PDLQWDLQHG LQ URRPV RQ D OLJKWUGDUN F\FOH )LVK ZHUH IHG 7HWUDPLQp IODNH IRRG RQFH HDFK GD\ $OO H[SHULPHQWDO DTXDULD ZHUH ORFDWHG LQ D FRQVWDQW WHPSHUDWXUH HQYLURQPHQW URRP WKDW PDLQWDLQHG DTXDULD DW s r& ([SHULPHQWDO DTXDULD ZHUH XVHG WR DFFOLPDWH ILVK WR VDOLQLWLHV UDQJLQJ IURP SSW WR SSW WR P2VP NJrOf 7KH LQLWLDO DFFOLPDWLRQ SHULRG ZDV GD\V DIWHU ZKLFK ILVK ZHUH HLWKHU XVHG LQ D PHWDEROLF WULDO RU ZHUH WUDQVIHUUHG WR WKH QH[W KLJKHU RU ORZHU VDOLQLW\ LQ WKH VHULHV 6DOLQLW\ FKDQJHV ZHUH LQ VWHSV RI SSW ZLWK VPDOOHU LQFUHPHQWV XVHG WR DFFOLPDWH ILVK WR SSW 7KLV SURFHGXUH ZDV UHSHDWHG XQWLO GHWHUPLQDWLRQV KDG EHHQ PDGH DW DOO H[SHULPHQWDO VDOLQLWLHV :DWHU XVHG LQ WKH IUHVKZDWHU DFFOLPDWLRQ ZDV REWDLQHG IURP ZHOOV LQ $ODFKXD DQG /HY\ FRXQWLHV )ORULGD PHDQ FRQGXFWLYLW\ S6 FPfOf ([SHULPHQWDO VDOLQLWLHV JUHDWHU WKDQ IUHVKZDWHU EXW OHVV WKDQ IXOO VHDZDWHU ZHUH SUHSDUHG E\ GLOXWLQJ ILOWHUHG $WODQWLF 2FHDQ VHDZDWHU REWDLQHG IURP WKH & 9 :KLWQH\ /DERUDWRU\ RI WKH 8QLYHUVLW\ RI )ORULGD 0DULQFODQG )ORULGDf ZLWK DSSURSULDWH TXDQWLWLHV RI GHLRQL]HG ZDWHU 6DOLQLWLHV JUHDWHU WKDQ SSW ZHUH SURGXFHG E\ VXSSOHPHQWLQJ VHDZDWHU ZLWK DSSURSULDWH

PAGE 26

DPRXQWV RI V\QWKHWLF VHD VDOW ,QVWDQW 2FHDQpf 6DOLQLWLHV ZHUH PRQLWRUHG GDLO\ ZLWK DQ $2p WHPSHUDWXUHFRPSHQVDWHG UHIUDFWRPHWHU DQG DGMXVWHG DV QHFHVVDU\ 0HWDEROLF GHWHUPLQDWLRQV ZHUH FDUULHG RXW LQ VHDOHG RSDTXH IODVNV UDQJLQJ LQ YROXPH IURP WR / ZLWK WKH YROXPH VHOHFWHG EDVHG RQ WKH VL]H RI WKH ILVK EHLQJ WHVWHG 5DWH RI R[\JHQ FRQVXPSWLRQ ZDV XVHG WR PHDVXUH PHWDEROLVP ZLWK IODVNV EHLQJ XVHG DV FORVHG UHVSLURPHWHUV 0HDVXUHV RI PHWDEROLVP ZHUH FRQVLGHUHG WR EH URXWLQH PHWDEROLF UDWHV DV DQLPDOV ZHUH VHTXHVWHUHG LQ VXFK D ZD\ DV WR PLQLPL]H EXW QRW HOLPLQDWH DFWLYLW\ :LQEHUJ )U\ f ([SHULPHQWV ZHUH SHUIRUPHG ZLWK SRVWDEVRUSWLYH ILVK LQ D UHVWLQJ VWDWH EXW ILVK ZHUH XQFRQVWUDLQHG DQG FDSDEOH RI VSRQWDQHRXV PRWRU DFWLYLW\ :LQEHUJ )U\ f )RU D UHODWLYHO\ LQDFWLYH VSHFLHV VXFK DV & YDULHJDWXV FRPSDUHG WR DFWLYHO\ VZLPPLQJ VDOPRQLGV IRU H[DPSOHf 505 LV SHUKDSV WKH PRVW DSSURSULDWH PHDVXUH RI PHWDEROLVP DV LW PRUH FORVHO\ UHIOHFWV QRUPDO DFWLYLW\ SDWWHUQV RI WKH ILVK WKDQ GRHV HLWKHU DFWLYH RU VWDQGDUG PHWDEROLF UDWH (DFK IODVN ZDV VHDOHG ZLWK D UXEEHU VWRSSHU WKURXJK ZKLFK WZR K\SRGHUPLF QHHGOHV 1R f ZHUH LQVHUWHG (DFK QHHGOH ZDV ILWWHG RQ WKH LQVLGH ZLWK FDWKHWHU WXELQJ RQH ZLWK D SLHFH ORQJ HQRXJK WR UHDFK WKH ERWWRP RI WKH IODVN WKH RWKHU KDOI WKLV OHQJWK $ PO SODVWLF V\ULQJH ILOOHG ZLWK ZDWHU DW WKH VDOLQLW\ RI WKH H[SHULPHQWDO DTXDULXP ZDV LQVHUWHG LQWR WKH QHHGOH ILWWHG ZLWK WKH VKRUW SLHFH RI FDWKHWHU WXELQJ ZKLOH DQ HPSW\ PO V\ULQJH ZDV LQVHUWHG LQWR WKH RWKHU QHHGOH $ PO ZDWHU VDPSOH ZDV GUDZQ LQWR WKH HPSW\ V\ULQJH IRU HDFK GHWHUPLQDWLRQ RI 3&! $V HDFK ZDWHU VDPSOH ZDV ZLWKGUDZQ DQ HTXDO YROXPH RI ZDWHU ZDV LQMHFWHG IURP WKH ILOOHG V\ULQJH LQWR WKH IODVN 7KLV PHWKRG RI VDPSOLQJ DORQJ ZLWK PLQRU PRYHPHQWV E\ WKH ILVK HIIHFWLYHO\ VWLUUHG WKH ZDWHU LQ WKH IODVN 1RUGOLH SHUV FRPPf 'HWHUPLQDWLRQV RI 32 ZHUH PDGH ZLWK D 5DGLRPHWHUp R[\JHQ HOHFWURGH FRQQHFWHG WR D 5DGLRPHWHUp 3+0 DFLGEDVH DQDO\]HU 0HDVXUHPHQWV RI WKH UDWH RI UHGXFWLRQ LQ 32 ZHUH PDGH DW K LQWHUYDOV DQG FRQWLQXHG XQWLO ILVK KDG GHSOHWHG WKH R[\JHQ OHYHO WR DSSUR[LPDWHO\ PP +J JHQHUDOO\ WR Kf )ROORZLQJ WKH ILQDO 32 GHWHUPLQDWLRQ HDFK ILVK ZfDV UHPRYHG IURP LWV IODVN GDPS

PAGE 27

GULHG DQG ZHLJKHG WR WKH QHDUHVW J $OO PHWDEROLF GHWHUPLQDWLRQV ZHUH PDGH EHWZHHQ DQG KRXUV DQG ILVK ZHUH QRW UHXVHG LQ RWKHU PHWDEROLF WULDOV 7R HQVXUH WKDW ILVK ZHUH SRVWDEVRUSWLYH DW WKH WLPH RI WHVWLQJ IRRG ZDV ZLWKKHOG IURP H[SHULPHQWDO DTXDULD IRU K SULRU WR EHJLQQLQJ D PHWDEROLF UHDGLQJ 5HVSLURPHWHUV ZHUH ILOOHG ZLWK ZDWHU DW WKH VDOLQLW\ RI WKH H[SHULPHQWDO DTXDULD DQG SODFHG LQ D ZDWHU EDWK PDLQWDLQHG DW s r& 7KH HQWLUH PHWDEROLF DSSDUDWXV ZDV ORFDWHG LQ D VPDOO VHPL GDUNHQHG URRP LQ ZKLFK QR RWKHU DFWLYLW\ WRRN SODFH ,Q RUGHU WR DOORZ WLPH IRU WKH ILVK WR DGMXVW WR WKH UHVSLURPHWHUV LQGLYLGXDOV ZHUH SODFHG LQWR WKH IODVNV ZLWK FRQVWDQW DHUDWLRQf WR K EHIRUH EHJLQQLQJ D WULDO $W WKH EHJLQQLQJ RI WKH PHWDEROLF WULDO DHUDWRUV ZHUH UHPRYHG DQG WKH IODVN ZDV VHDOHG &DOFXODWLRQ RI R[\JHQ VDWXUDWLRQ YDOXHV IRU WKH H[SHULPHQWDO FRQGLWLRQV WDNLQJ LQWR DFFRXQW VDOLQLW\ WHPSHUDWXUH UHODWLYH KXPLGLW\ DQG EDURPHWULF SUHVVXUHf ZHUH PDGH XVLQJ WKH HTXDWLRQV RI 7UXHVGDOH HW DO DQG ERWK 505 PJ KfOf DQG 3F PP +Jf ZHUH FDOFXODWHG IRU HDFK ILVK 'DWD XVHG IRU FDOFXODWLRQ RI PHWDEROLF UDWHV ZHUH OLPLWHG WR YDOXHV REWDLQHG ZKLOH WKH 32R LQ WKH UHVSLURPHWHU ZDV JUHDWHU WKDQ PP +J LQ RUGHU WR HQVXUH WKDW WKHVH FDOFXODWLRQV ZHUH PDGH DW R[\JHQ WHQVLRQV ZHOO DERYH WKH 3F $OO GDWD ZHUH XVHG IRU FDOFXODWLRQ RI WKH 3F 'HWHUPLQDWLRQ RI WKH 3F ZDV PDGH XVLQJ D %$6,& SURJUDP WR FDOFXODWH WKH FULWLFDO SRLQW
PAGE 28

GHVFULELQJ WKH UHODWLRQVKLS EHWZHHQ PDVV DQG PHWDEROLVP IRU & YDULHJDWXV 1RUGOLH HW DO f 05 N: ZDV XVHG WR FRUUHFW R[\JHQ FRQVXPSWLRQ UDWHV 9DOXHV ZHUH FRUUHFWHG IROORZLQJ WKH UHODWLRQVKLS 05F :Dfrf05Rf ZKHUH 05F LV WKH PDVV FRUUHFWHG R[\JHQ FRQVXPSWLRQ : LV WKH REVHUYHG PDVV DQG 05R LV WKH REVHUYHG R[\JHQ FRQVXPSWLRQ DW PDVV :4 8OWVFK HW DO &HFK f 6WDWLVWLFDO DQDO\VHV IROORZ SURFHGXUHV RXWOLQHG LQ :LQHU HW DO f DQG 6RNDO DQG 5RKOI f $OO VWDWLVWLFDO DQDO\VHV ZHUH RQH ZD\ WHVWV XVLQJ WKH 7XNH\.UDPHU SRVW KRF FRPSDULVRQ S f DQG YDOXHV DUH JLYHQ WKURXJKRXW DV PHDQV s VWDQGDUG HUURU RI WKH PHDQ VHf )LHOG 0HDVXUHPHQWV 6DOW PDUVK KDELWDWV DUH ZLGHO\ FRQVLGHUHG WR H[SHULHQFH XQSUHGLFWDEOH DQG IOXFWXDWLQJ DELRWLF FRQGLWLRQV +RZHYHU DFWXDO SK\VLFRFKHPLFDO PHDVXUHPHQWV DUH LQIUHTXHQWO\ UHSRUWHG 7R DGGUHVV WKLV LVVXH ILHOG PHDVXUHPHQWV ZHUH PDGH DW IRXU VLWHV LQ WKH &HGDU .H\ DUHD RYHU D RQH \HDU SHULRG :KHQHYHU SRVVLEOH PHDVXUHPHQWV DW HDFK VLWH ZHUH WDNHQ ERWK DW WKH VXUIDFH DQG RQ WKH ERWWRP JHQHUDOO\ P GHHSf 'LVVROYHG R[\JHQ VDOLQLW\ DQG WHPSHUDWXUH ZHUH PHDVXUHG RQH WR WKUHH WLPHV HDFK PRQWK EHWZHHQ K DQG K IRU D WRWDO RI GDWHV EHWZHHQ -XQH DQG -XQH 6LWHV DQG ZHUH ORFDWHG GHHS LQ WKH VDOW PDUVK ZKHUH & YDULHJDWXV ZDV URXWLQHO\ FROOHFWHG 7KHVH VLWHV ZHUH ORFDWHG LQ FORVH SUR[LPLW\ WR RQH DQRWKHU P DSDUWf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

PAGE 29

H[SHULHQFHG E\ VDOW PDUVK LQKDELWDQWV DQG JLYH DQ LQGLFDWLRQ RI VRPH RI WKH YDULDELOLW\ LQ WKH PHDVXUHG SK\VLFRFKHPLFDO SDUDPHWHUV 5HVXOWV 5RXWLQH 0HWDEROLVP 0HDQ 505 ZDV FDOFXODWHG IRU HDFK ILVK DQG RUJDQL]HG E\ VDOLQLW\ JURXSV 0HDQ YDOXHV IRU XQDGMXVWHG DQG DGMXVWHG 505 IURP $1&29$f DUH JLYHQ LQ 7DEOH DQG DGMXVWHG 505 DUH SORWWHG DJDLQVW DPELHQW VDOLQLW\ LQ )LJXUH ,Q WKH UDQJH RI DPELHQW VDOLQLWLHV EHWZHHQ IUHVKZDWHU DQG SSW DGMXVWHG 505 YDOXHV ZHUH KLJKHVW DW SSW DQG SSW EHLQJ VOLJKWO\ ORZHU DQG URXJKO\ HTXLYDOHQW DW WKH RWKHU PHDVXUHG VDOLQLWLHV LQ WKLV UDQJH $W VDOLQLWLHV JUHDWHU WKDQ SSW WKHUH ZDV D SURJUHVVLYH GHFOLQH LQ DGMXVWHG 505 2YHUDOO DGMXVWHG 505 UDQJHG IURP D PD[LPXP RI PJ KfO DW SSW WR D ORZ RI PJ KnO DW SSW UHSUHVHQWLQJ D b GHFOLQH 7KLV GHFOLQH FRUUHVSRQGV ZLWK D GHFUHDVHG DELOLW\ WR UHJXODWH SODVPD RVPRODOLW\ DW HOHYDWHG VDOLQLWLHV )LJXUH SODVPD RVPRODOLW\ GDWD DUH IURP 1RUGOLH f $ PXOWLSOH OLQHDU UHJUHVVLRQ DQDO\VLV ZDV XVHG WR JHQHUDWH D SUHGLFWLYH PRGHO IRU GHVFULELQJ WKH HIIHFWV RI ERWK VDOLQLW\ DQG PDVV RQ PHWDEROLVP ,Q WKLV PRGHO /RJ ERG\ PDVV /RJ : LQ Jf DQG VDOLQLW\ 6 LQ SSWf ZHUH XVHG DV LQGHSHQGHQW YDULDEOHV DQG /RJ PDVVLQGHSHQGHQW 505 /RJ 05 LQ PJ 2 KnOf DV WKH GHSHQGHQW YDULDEOH 7KH HTXDWLRQ WKDW EHVW GHVFULEHG WKLV UHODWLRQVKLS LV /RJ 05 /RJ :f 6f )RO 3 f 7KLV PRGHO GHVFULEHG b RI DOO YDULDELOLW\ DERXW WKH PHDQ DQG WKH UDQGRP GLVWULEXWLRQ RI WKH UHVLGXDOV VXJJHVWV DQ DEVHQFH RI VLJQLILFDQW UHODWLRQVKLSV WKDW PLJKW KDYH ELDVHG WKH DQDO\VLV

PAGE 30

7DEOH 5HODWLRQVKLSV RI URXWLQH PHWDEROLVP 505f FULWLFDO R[\JHQ WHQVLRQ 3Ff DQG VORSH LQ WKH FRQIRUPDWLRQ UHJLRQ DW D VHULHV RI DPELHQW VDOLQLWLHV 9DOXHV DUF JLYHQ DV PHDQV s VH $PELHQW 6DOLQLW\ 332 Q 0HDQ %RG\ 0DVV Jf 8QDGMXVWHG 505 PJ &! KnOf $GMXVWHG 505 PJ Knf &ULWLFDO 2[\JHQ 7HQVLRQ PP +Jf 6ORSH LQ &RQIRUPDWLRQ 5HJLRQ s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s

PAGE 31

)LJXUH 0HDQ DGMXVWHG URXWLQH PHWDEROLF UDWHV 505f RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YFLULHJDWXV PHWDEROLF UDWHV ZHUH PDVV DGMXVWHG XVLQJ DQ DQDO\VLV RI FRYDULDQFH EDUV LQGLFDWH s VH QXPHULFDO YDOXHV DERYH WKH SRLQWV LQ WKH ILJXUH LQGLFDWH VDPSOH VL]H DW HDFK VDOLQLW\f

PAGE 32

6DOLQLW\ SSWf $GMXVWHG 5RXWLQH 0HWDEROLF 5DWH PJ 2[\JHQ KU f SRRS I U Lr E? ER A Lr ?=

PAGE 33

)LJXUH 5HODWLRQVKLS EHWZHHQ PHDQ DGMXVWHG URXWLQH PHWDEROLF UDWHV 505f DQG PHDQ SODVPD RVPRODOLW\ RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YDULHJDWXV PHWDEROLF UDWHV ZHUH PDVVDGMXVWHG XVLQJ DQ DQDO\VLV RI FRYDULDQFH EDUV LQGLFDWH s VH SODVPD RVPRODOLW\ GDWD IURP 1RUGOLH f

PAGE 34

$GMXVWHG 5RXWLQH 0HWDEROLF 5DWH PJ 2[\JHQKUf 3ODVPD 2VPRODOLW\ P2VPNJf

PAGE 35

&ULWLFDO 2[\JHQ 7HQVLRQ 0HDVXUHPHQWV RI R[\JHQ FRQVXPSWLRQ PJ KnOf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f VLPLODU WR WKH SDWWHUQ H[KLELWHG E\ WKH 505 GDWD 0HDQ 3F YDOXHV LQFUHDVHG DW VDOLQLWLHV JUHDWHU WKDQ SSW ZLWK WKH KLJKHVW OHYHOV UHFRUGHG DW VDOLQLWLHV SSW DQG KLJKHU 3F YDOXHV UDQJHG IURP D ORZ RI PP +J DW D VDOLQLW\ RI SSW WR D KLJK RI PP +J DW SSW UHSUHVHQWLQJ D b LQFUHDVH 7KH ULVH LQ PHDQ 3F YDOXHV FRUUHVSRQGV ZHOO ZLWK D GHFUHDVHG DELOLW\ WR UHJXODWH SODVPD RVPRODOLW\ DJDLQ VLPLODU WR WKH 505 SDWWHUQ )LJXUH SODVPD RVPRODOLW\ GDWD DUH IURP 1RUGOLH f %HORZ WKH 3F PHWDEROLVP GHSHQGV RQ WKH R[\JHQ WHQVLRQ DQG GHFUHDVHV DV WKH 32 GHFUHDVHV 0DWKHPDWLFDOO\ WKH VORSH RI WKH UHVXOWLQJ OLQH LQ WKLV FRQIRUPDWLRQ UHJLRQ GHSHQGV RQ WKUHH IDFWRUV WKH 3F WKH 505 DW WKH 3F DQG WKH OHWKDO 3 $ FRPSDULVRQ RI WKH VORSHV LQ WKH FRQIRUPDWLRQ UHJLRQ UHYHDOV WKDW GHVSLWH WKH FKDQJHV QRWHG DERYH LQ 505 DQG 3F YDOXHV WKH 32 DW ZKLFK WKH ILVK FDQ QR ORQJHU VXUYLYH XQGHU H[SHULPHQWDO FRQGLWLRQVf LV HVVHQWLDOO\ HTXLYDOHQW IRU DOO VDOLQLWLHV WHVWHG 7KLV LV UHIOHFWHG LQ WKH LQFUHDVLQJO\ VKDOORZ VORSHV VHHQ DW VDOLQLWLHV JUHDWHU WKDQ SSW 7DEOH f )LHOG 0HDVXUHPHQWV 7KH ILHOG PHDVXUHPHQWV RI R[\JHQ FRQFHQWUDWLRQ VDOLQLW\ DQG WHPSHUDWXUH UHYHDOHG YHU\ KLJK YDULDELOLW\ RI WKHVH SK\VLFRFKHPLFDO SDUDPHWHUV RYHU WKH FRXUVH RI WKH VDPSOLQJ

PAGE 36

)LJXUH 0HDQ FULWLFDO R[\JHQ WHQVLRQ 3Ff PHDVXUHPHQWV RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YDULHJFLWXV EDUV LQGLFDWH s VH QXPHULFDO YDOXHV DERYH WKH SRLQWV LQ WKH ILJXUH LQGLFDWH VDPSOH VL]H DW HDFK VDOLQLW\f

PAGE 37

6DOLQLW\ SSWf &ULWLFDO 2[\JHQ 7HQVLRQ PP +Jf ‘X WQ R? RR YR R R R R R R F

PAGE 38

)LJXUH 5HODWLRQVKLS EHWZHHQ PHDQ DGMXVWHG URXWLQH PHWDEROLF UDWHV 505f DQG FULWLFDO R[\JHQ WHQVLRQV 3Ff RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YDULHJDWXV PHWDEROLF UDWHV ZHUH PDVVDGMXVWHG XVLQJ DQ DQDO\VLV RI FRYDULDQFH EDUV LQGLFDWH s VHf

PAGE 39

$GMXVWHG 5RXWLQH 0HWDEROLF 5DWH PJ 2[\JHQKUf 6DOLQLW\ SSWf &ULWLFDO 2[\JHQ 7HQVLRQ PP +Jf

PAGE 40

)LJXUH 5HODWLRQVKLS EHWZHHQ PHDQ FULWLFDO R[\JHQ WHQVLRQV 3Ff DQG PHDQ SODVPD RVPRODOLW\ RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YDULHJDWXV EDUV LQGLFDWH s VH SODVPD RVPRODOLW\ GDWD IURP 1RUGOLH f

PAGE 41

&ULWLFDO 2[\JHQ 7HQVLRQ PP +Jf &ULWLFDO 2[\JHQ 7HQVLRQ 3ODVPD 2VPRODOLW\ ? n } L ‘ U 6DOLQLW\ SSWf 3ODVPD 2VPRODOLW\ P2VPNJf

PAGE 42

SHULRG 7DEOH f )RU DOO VLWHV FRPELQHG UDQJHV RI GLVVROYHG R[\JHQ VDOLQLW\ DQG WHPSHUDWXUH RYHU WKH FRXUVH RI WKH \HDU ZHUH PJ /nO SSW DQG r& UHVSHFWLYHO\ (YHQ KLJKHU VDOLQLWLHV SSWf ZHUH HQFRXQWHUHG RFFDVLRQDOO\ LQ WKH VDOW PDUVK RXWVLGH WKH VDPSOLQJ SHULRG 2WKHU DUHDV RI WKH VDOW PDUVK SUREDEO\ HQFRXQWHU HYHQ JUHDWHU H[WUHPHV RI WKHVH YDULDEOHV &\SULQRGRQ YDULHJDWXV ZDV VHHQ RQ DOO GDWHV ZKHQ SK\VLFRFKHPLFDO PHDVXUHPHQWV ZHUH PDGH ,Q DGGLWLRQ WR KLJK RYHUDOO YDULDELOLW\ WKHUH ZHUH ODUJH GLIIHUHQFHV DPRQJ VRPH VLWHV LQ FORVH SUR[LPLW\ WR RQH DQRWKHU )RU H[DPSOH PHDQ R[\JHQ FRQFHQWUDWLRQ PHDVXUHPHQWV WDNHQ GXULQJ WKH )DOO DW VLWHV RQH WZR DQG WKUHH ZHUH PJ /f PJ / DQG PJ /nO UHVSHFWLYHO\ $OO WKUHH SK\VLFRFKHPLFDO SDUDPHWHUV DOVR VWURQJO\ YDULHG WHPSRUDOO\ DPRQJ VDPSOLQJ GDWHV 7KHVH GDWD SURYLGH JRRG HYLGHQFH WKDW WKH &HGDU .H\ VDOW PDUVK LV DQ H[WUHPHO\ YDULDEOH KDELWDW ZLWK UHVSHFW WR WKHVH SK\VLFRFKHPLFDO SDUDPHWHUV 'LVFXVVLRQ 7KH IDPLO\ &\SULQRGRQWLGDH LV D GLYHUVH JURXS RI ILVKHV ZLWK PDQ\ VSHFLHV WKDW WROHUDWH H[WUHPH HQYLURQPHQWDO FRQGLWLRQV /RZH HW DO /RWDQ DQG 6NDGKDXJH 1DLPDQ HW DO 6WXHQNHO DQG +LOO\DUG &KXQJ f &\SULQRGRQ YDULHJDWXV LV SHUKDSV WKH PRVW SK\VLRORJLFDOO\ WROHUDQW PHPEHU RI WKH IDPLO\ ,W KDV EHHQ FDOOHG WKH WRXJKHVW ILVK LQ 1RUWK $PHULFD *XQWHU f GXH WR LWV H[WUHPH WROHUDQFH RI VHYHUH DELRWLF FRQGLWLRQV 7KH VSHFLHV LV IRXQG LQ ZDWHUV UDQJLQJ LQ VDOLQLW\ IURP IUHVKZDWHU $JHU f WR SSW 6LPSVRQ DQG *XQWHU f DQG FDQ UHSURGXFH LQ ZDWHUV DV KLJK DV SSW 0DUWLQ f 7KH\ DUH NQRZQ WR WROHUDWH WHPSHUDWXUHV UDQJLQJ IURP DERXW r& %HUU\ f WR WHPSHUDWXUHV JUHDWHU WKDQ r& 6WUDZQ DQG 'XQQ f DQG WR WROHUDWH QHDU DQR[LF FRQGLWLRQV 2GXP DQG &DOGZHOO f 7KXV WKLV VSHFLHV LV DQ H[FHHGLQJO\ XVHIXO H[SHULPHQWDO VXEMHFW IRU H[DPLQLQJ KRZ WFOFRVWV UHVSRQG WR KDUVK HQYLURQPHQWDO FRQGLWLRQV

PAGE 43

7DEOH 0HDVXUHPHQWV RI R[\JHQ FRQFHQWUDWLRQ PJ /nOf VDOLQLW\ SSWf DQG WHPSHUDWXUH r&f WDNHQ DW IRXU VLWHV LQ WKH &HGDU .H\ DUHD IURP -XQH WKURXJK -XQH VHH WH[W IRU GHWDLOV RQ ORFDWLRQ RI VLWHVf 9DOXHV DUH JLYHQ DV PHDQV VH LQ SDUHQWKHVHVf VDPSOH VL]H 2[\JHQ PJ /Of 6DOLQLW\ SSWf 7HPSHUDWXUH r&f /RFDWLRQ 'HSWK 6SULQJ 6XPPHU )DOO :LQWHU 6SULQJ 6XPPHU )DOO :LQWH U 6SULQJ 6XPPHU )DOO :LQWHU 6LWH %RWWRP f f f f f f f f f f f f Q Q Q Q Q Q Q Q Q Q Q Q 6XUIDFH f f f f f f f f f f f f Q Q Q Q Q Q Q Q Q Q Q Q 6LWH %RWWRP f f f f f f f f f f f f Q Q Q Q Q Q Q Q Q Q Q Q 6XUIDFH f f f f f f f f f f f f Q Q Q Q Q Q Q Q Q Q Q Q 6LWH %RWWRP f f f f f f f f f f f f Q Q Q Q Q Q Q Q Q Q Q Q f§ f§ f§ 6XUIDFH f f f f§ f f f f§ f f f f§ Q Q Q f§ Q Q Q f§ Q Q Q f§ 6LWH 0LGGOH f f f f f f f f f f f f Q Q Q Q Q Q Q Q Q Q Q Q

PAGE 44

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f DV FDWHJRUL]HG E\ ERWK 1RUGOLH f DQG 0RUJDQ DQG ,ZDPD f $ VLPLODU SDWWHUQ FKDUDFWHUL]HV WKH UHODWHG VDOW PDUVK UHVLGHQW $GLQLD [HQLFD '& +DQH\ XQSXEOLVKHG GDWDf LQ ZKLFK 505 ZDV PRUH YDULDEOH EXW UHODWLYHO\ FRQVWDQW RYHU VDOLQLWLHV IURP SSW WR SSW )LJXUH f &\SULQRGRQ YDULHJDWXV H[KLELWV D GHFOLQH LQ 505 RQO\ DW VDOLQLWLHV H[FHHGLQJ SSW 7KLV UHVXOW ZDV VRPHZKDW XQH[SHFWHG EXW LV QHDUO\ LGHQWLFDO WR WKH SDWWHUQ IRXQG E\ 1RUGOLH HW DO f LQ D VWXG\ WKDW DOVR H[DPLQHG PHWDEROLVP RI & YDULHJDWXV RYHU D ZLGH UDQJH RI DPELHQW VDOLQLWLHV 1RUGOLH HW DO f FRQFOXGHG WKDW WKH GHSUHVVLRQ LQ PHWDEROLVP DW KLJK VDOLQLWLHV LV SUREDEO\ UHODWHG WR SHUPHDELOLW\ FKDQJHV RI WKH JLOO PHPEUDQH DQGRU LQWHJXPHQW 7KH SRLQW DW ZKLFK PHWDEROLVP LV UHGXFHG FRUUHVSRQGV ZHOO ZLWK D GLPLQLVKHG DELOLW\ RI & YDULHJDWXV WR RVPRUHJXODWH HIILFLHQWO\ ,I RVPRWLF SHUPHDELOLW\ RI WKH JLOOV LV UHGXFHG DW KLJK VDOLQLWLHV WR KHOS RIIVHW LRQLF LQIOX[ DQG RVPRWLF HIIOX[ WKH SRWHQWLDO IRU R[\JHQ XSWDNH PD\ EH UHGXFHG DV ZHOO .ULVWHQVHQ DQG 6NDGKDXJH 6NDGKDXJH 'DYHQSRUW DQG 6D\FU f (YLGHQFH IRU WKLV K\SRWKHVLV FRPHV IURP VHYHUDO UHFHQW VWXGLHV ,Q WKH ILUVW RI WKHVH .XOW] DQG 2QNHQ f IRXQG WKDW RYHUDOO LQ YLWUR SHUPHDELOLW\ RI WKH RSHUFXODU PHPEUDQH RI WKH FLFKOLG 2UHRFKURPLV PRVVDPELFXV ZDV UHGXFHG LQ K\SFUVDOLQH PHGLD ZLWK D VLPXOWDQHRXV UHGXFWLRQ LQ SDVVLYH LRQ IOX[HV 0RUH GLUHFW HYLGHQFH FRPHV IURP VWXGLHV E\ %LQGRQ HW DO Df DQG %LQGRQ HW DO Ef RQ WKH UDLQERZ WURXW 2QFRUK\QFKXV P\NLVV ,Q WKHVH VWXGLHV WKH DXWKRUV GHPRQVWUDWHG WKDW LPSDLUPHQW RI UHVSLUDWRU\ JDV WUDQVIHU FRLQFLGHG ZLWK FKORULGH FHOO SUROLIHUDWLRQ LQGXFHG E\ DQ RVPRUHJXODWRU\ FKDOOHQJH

PAGE 45

)LJXUH 5HODWLRQVKLS EHWZHHQ PHDQ DGMXVWHG URXWLQH PHWDEROLF UDWHV 505f DQG PHDQ SODVPD RVPRODOLW\ RYHU D UDQJH RI VDOLQLWLHV LQ $GLQLD [HQLFD PHWDEROLF UDWHV ZHUH PDVVDGMXVWHG XVLQJ DQ DQDO\VLV RI FRYDULDQFH EDUV LQGLFDWH s VH QXPHULFDO YDOXHV DERYH WKH SRLQWV LQ WKH ILJXUH LQGLFDWH VDPSOH VL]H DW HDFK VDOLQLW\f

PAGE 46

$GMXVWHG 5RXWLQH 0HWDEROLF 5DWH 0J 2[\JHQKUf 6DOLQLW\ SSWf 3ODVPD 2VPRODOLW\ P2VP.Jf

PAGE 47

&KDQJHV LQ DFWLYLW\ PD\ DOVR DFFRXQW IRU D GHSUHVVLRQ LQ PHWDEROLVP ,Q D VWXG\ RQ WKH PLONILVK &KDQRV FKDQRV 6ZDQVRQ f VKRZHG WKDW D GHSUHVVLRQ LQ PHWDEROLVP DW DQ HOHYDWHG VDOLQLW\ SSWf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f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f H[DPLQHG R[\JHQ FRQVXPSWLRQ RI 7LODSLD PRVVDPELFD 2 PRVVDPELFXVf DW SSW SSW DQG SSW DQG IRXQG QR HIIHFW RI VDOLQLW\ RQ WKH 3F RI VPDOO Jf ILVK DW HLWKHU r& r& RU r& 3F DSSUR[LPDWHO\ PP +Jf 6DOLQLW\ GLG DIIHFW 3F RQ ODUJHU Jf LQGLYLGXDOV DW r& ZKHUH WKH 3F GRXEOHG IURP DSSUR[LPDWHO\ PP +J WR PP +J LQ ILVK DFFOLPDWHG WR SSW YHUVXV SSW RU SSW +RZHYHU 3F FDOFXODWLRQV PDGH E\ -RE f ZHUH H[WUHPHO\ URXJK DSSUR[LPDWLRQV DQG DUH GLIILFXOW WR FRPSDUH ZLWK YDOXHV LQ WKLV VWXG\

PAGE 48

6XEUDKPDQ\DP f H[DPLQHG WKH LQIOXHQFH RI R[\JHQ WHQVLRQ RQ WKH PHWDEROLF UDWH RI VHYHUDO VDOW PDUVK ILVKHV LQFOXGLQJ & YDULHJDWXV ZLWK DOO PHDVXUHPHQWV PDGH DW r& DQG VDOLQLWLHV RI SSW $OO VSHFLHV WHVWHG LQ 6XEUDKPDQ\DP f & YDULHJDWXV 3RHFLOLD ODWLSLQQD /DJRGRQ UKRPERLGHV /HLRVWRUQXV [DQWKXUXV )XQGXOXV JUDQ£LV DQG ) VLPLOLVf ZHUH R[\JHQ FRQIRUPHUV DW R[\JHQ WHQVLRQV RI PP +J DQG ORZHU D VRPHZKDW GLIIHUHQW UHVSRQVH WKDQ VHHQ LQ WKLV VWXG\ 7KLV GLIIHUHQFH PD\ EH GXH WR WKH VPDOO VDPSOH VL]HV DQG UHODWLYHO\ IHZ PHDVXUHPHQWV RI R[\JHQ FRQVXPSWLRQ PDGH E\ 6XEUDKPDQ\DP f ,Q D VWXG\ RQ WKH JROGHQ PXOOHW /L]D DXUDWD 6KXVPLQ f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f WR WR PP +J LQ *RELXV FRELWXV %ULGJHV f DQG +HOFRJUDPPD PHGLXP ,QQHV DQG :HOOV 3HOVWHU HW DO 4XLQQ DQG 6FKQHLGHU f $ QXPEHU RI $IULFDQ FLFKOLGV HJ 2UHRFKURPLV QLORWLFXV &LFKODVRPD XURSKOKDODPXV (UHOPRGXV F\DQRVWLFXV 'LPLGLRFKURPLV FRPSUHVVLFHSVf KDYH HLWKHU VOLJKWO\ ORZHU RU URXJKO\ HTXLYDOHQW 3F YDOXHV WR WKH DSSUR[LPDWH YDOXH PP +Jf GLVSOD\HG E\ & YDULHJDWXV LQ WKLV VWXG\ DW PRVW VDOLQLWLHV 5RVV DQG 5RVV %HFNHU DQG )LVKHOVRQ 3DODFLRV DQG 5RVV 9HUKF\FQ FW DO f 'RQQHOO\ DQG 7RUUHV f IRXQG 3F YDOXHV UDQJLQJ IURP WR PP +J IRU D QXPEHU RI PLGZDWFU ILVKHV IURP WKH HDVWHUQ *XOI RI 0H[LFR DJDLQ YDOXHV QHDU RU VOLJKWO\ ORZHU WKDQ WKRVH RI & YDULHJDWXV 7KXV & YDULHJDWXV KDV 3F YDOXHV WKDW DUF IDLUO\ FORVH WR WKRVH RI HFRORJLFDOO\ DQG HYROXWLRQDULO\ GLYHUVH WFOFRVWV

PAGE 49

7KH YDULDWLRQV LQ 505 DQG 3F DV D IXQFWLRQ RI HQYLURQPHQWDO VDOLQLW\ REVHUYHG LQ WKLV VWXG\ VXJJHVW WKDW & YDULHJDWXV UHVSRQGV WR KLJK VDOLQLWLHV E\ UHGXFLQJ HQHUJ\ H[SHQGLWXUHV 2EVHUYDWLRQV E\ P\VHOI DQG RWKHUV LQGLFDWH WKDW K\SHUVDOLQH FRQGLWLRQV DUH HQFRXQWHUHG LQIUHTXHQWO\ DQG OLNHO\ ODVW IRU VKRUW GD\Vf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f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

PAGE 50

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f &RPSDUDWLYHO\ IHZHU VWXGLHV KDYH EHHQ FRQVLGHUHG WKH LQIOXHQFH RI VDOLQLW\ RQ PHWDEROLF SDWWHUQV RI ILVKHV HJ .LQQH 0DGDQ 0RKRQ 5DR 1RUGOLH HW DO 6ZDQVRQ f ,Q WKHVH VWXGLHV YDULRXV PHWDEROLF UHVSRQVHV WR VDOLQLW\ KDYH EHHQ UHSRUWHG (XU\KDOLQH ILVK H[KLELW RQH RI WKH IHZ FRQVLVWHQW SDWWHUQV QDPHO\ WKDW PHWDEROLVP UHPDLQV UHODWLYHO\ XQDIIHFWHG RYHU WKH UDQJH RI VDOLQLWLHV QRUPDOO\ HQFRXQWHUHG 1RUGOLH 0RUJDQ DQG ,ZDPD f 0RVW VWXGLHV WKDW KDYH H[DPLQHG WKH LQIOXHQFH RI VDOLQLW\ RQ ILVK HQHUJHWLFV KDYH HPSOR\HG PHDVXUHPHQWV IURP ILVK PDLQWDLQHG DW FRQVWDQW VDOLQLWLHV 'DWD IURP WKHVH H[SHULPHQWV DUH YHU\ XVHIXO IRU XQGHUVWDQGLQJ WKH RYHUDOO PHWDEROLF SDWWHUQV H[KLELWHG E\ WKH VSHFLHV WHVWHG DV ILVK ZHUH XVXDOO\ IXOO\ DFFOLPDWHG WR HDFK H[SHULPHQWDO VDOLQLW\ SULRU WR WHVWLQJ +RZHYHU WKHVH H[SHULPHQWV SURYLGH OHVV LQIRUPDWLRQ DERXW PRUH HFRORJLFDOO\

PAGE 51

UHOHYDQW UHVSRQVHV DV ILVK LQ WKHLU QDWLYH KDELWDWV DUH RIWHQ VXEMHFW WR UDSLG DQG GUDPDWLF IOXFWXDWLRQV LQ VDOLQLW\ 6WXGLHV RI WKH LQIOXHQFH RI VDOLQLW\ IOXFWXDWLRQV RQ PHWDEROLVP RI ILVKHV KDYH XVXDOO\ IRFXVHG RQ VSHFLHV WKDW JHQHUDOO\ H[SHULHQFH PD[LPDO IOXFWXDWLRQV LQ VDOLQLW\ RQO\ EHWZHHQ IUHVKZDWHU SSWf DQG VHDZDWHU SSWf HJ 'DYHQSRUW DQG 9DKO 9RQ 2HUW]HQ 0RVHU DQG *HUU\ 6KXVPLQ 0RVHU DQG 0LOOHU f +RZHYHU VRPH VDOW PDUVK WHOHRVWV UHJXODUO\ HQFRXQWHU VDOLQLWLHV RXWVLGH WKLV UDQJH 7KLV VWXG\ H[DPLQHG WKH LQIOXHQFH RI VDOLQLW\ IOX[HV RQ URXWLQH PHWDEROLF UDWH 505f RI WKH VDOW PDUVK WHOHRVW &\SULQRGRQ YDULHJDWXV D VSHFLHV WKDW UHJXODUO\ HQFRXQWHUV VDOLQLWLHV JUHDWHU WKDQ SSW )LVK XVHG LQ WKLV VWXG\ ZHUH IXOO\ DFFOLPDWHG WR D VHULHV RI VDOLQLWLHV UDQJLQJ IURP SSW WR SSW IROORZHG E\ H[SRVXUH WR D VLPXODWHG WLGDO FKDQJH LQ VDOLQLW\ 7KH PDJQLWXGH UDWH DQG GLUHFWLRQ RI VDOLQLW\ FKDQJHV PD\ EH LPSRUWDQW GHWHUPLQDQWV RI KRZ VDOLQLW\ IOXFWXDWLRQV DIIHFW PHWDEROLVP ,Q WKLV VWXG\ WKH GLUHFWLRQ RI WKH VDOLQLW\ FKDQJH LQ FRQMXQFWLRQ ZLWK WKH DPELHQW DFFOLPDWLRQf VDOLQLW\ ZDV WKH SULPDU\ IRFXV 7KH PDJQLWXGH RI WKH VDOLQLW\ FKDQJHV ZHUH VHOHFWHG WR VLPXODWH H[WUHPHV NQRZQ WR RFFXU LQ WKH KDELWDW IURP ZKLFK ILVK ZHUH FROOHFWHG DQG WKH UDWH ZDV FKRVHQ WR VLPXODWH D QRUPDO GLXUQDO WLGDO F\FOH SUHGLFWHG WKDW & YDULHJDWXV ZRXOG VKRZ PLQLPDO FKDQJHV LQ 505 ZKHQ WKH VDOLQLW\ ZDV FKDQJHG ZLWKLQ WKH UDQJH FRPPRQO\ HQFRXQWHUHG E\ WKH VSHFLHV 6DOLQLW\ FKDQJHV RXWVLGH WKLV UDQJH ZHUH K\SRWKHVL]HG WR OHDG WR GHSUHVVLRQV LQ PHWDEROLVP 0HWKRGV &ROOHFWLRQV RI ILVK WUDQVSRUWDWLRQ EDFN WR WKH ODERUDWRU\ DQG JHQHUDO ODERUDWRU\ SURFHGXUHV ZHUH SHUIRUPHG DV GHVFULEHG SUHYLRXVO\ )LVK ZHUH ILUVW VHTXHQWLDOO\ DFFOLPDWHG WR D VHULHV RI VDOLQLWLHV UDQJLQJ IURP SSW WR SSW WR P2VP NJnOf +RZHYHU XQOLNH WKH SUHYLRXV H[SHULPHQWV WKHVH SURFHGXUHV ZHUH GHVLJQHG WR PHDVXUH 505 EHIRUH

PAGE 52

DQG IROORZLQJ VLPXODWHG WLGDO FKDQJHV LQ VDOLQLW\ ,QLWLDO VDOLQLWLHV DFFOLPDWLRQf DQG VDOLQLWLHV IROORZLQJ WKH VLPXODWHG WLGDO FKDQJH ILQDOf DUH VKRZQ LQ 7DEOH 0HWDEROLF GHWHUPLQDWLRQV DQG FKDQJHV LQ VDOLQLW\ ZHUH FDUULHG RXW LQ D IORZWKURXJK UHVSLURPHWHU ZLWK DQ HIIHFWLYH YROXPH RI / )LJXUH f 7KH UHVSLURPHWHU ZDV LPPHUVHG LQ D WKHUPRUHJXODWHG UHVHUYRLU GXULQJ PHWDEROLF WULDOV WKDW VHUYHG WR PDLQWDLQ D FRQVWDQW WHPSHUDWXUH ZLWKLQ WKH UHVSLURPHWHU $ VXEPHUVLEOH SXPS PDLQWDLQHG ZLWKLQ D VHFRQG WKHUPRUHJXODWHG UHVHUYRLU /f ZDV FRQQHFWHG WR WKH LQOHW RI WKH UHVSLURPHWHU 7KLV UHVHUYRLU ZDV YLJRURXVO\ DHUDWHG DQG ZDV XVHG WR VXSSO\ ZDWHU WR WKH UHVSLURPHWHU 7KH UHVSLURPHWHU RXWOHW HPSWLHG EDFN LQWR WKLV PDLQ UHVHUYRLU $ WKLUG UHVHUYRLU /f VHUY LQJ DV D VDOLQLW\ VRXUFH ZDV FRQQHFWHG WR WKH PDLQ UHVHUYRLU YLD D SHULVWDOWLF SXPS 'XH WR OLPLWDWLRQV RI WKH DYDLODEOH HTXLSPHQW WKH V\VWHP ZDV FORVHG GXULQJ R[\JHQ SDUWLDO SUHVVXUH 32f GHWHUPLQDWLRQV 7ZR SRUWV ZLWKLQ WKH UHVSLURPHWHU ZHUH XVHG WR VDPSOH ZDWHU IRU GHWHUPLQDWLRQ RI 3 $ PO SODVWLF V\ULQJH ILOOHG ZLWK ZDWHU DW WKH H[SHULPHQWDO VDOLQLW\ ZDV LQVHUWHG LQWR WKH ILUVW SRUW LQOHW VLGHf ZKLOH DQ HPSW\ PO V\ULQJH ZDV ILWWHG LQWR WKH VHFRQG SRUW RXWOHW VLGHf $V D PO ZDWHU VDPSOH ZDV GUDZQ LQWR WKH HPSW\ V\ULQJH DQ HTXDO YROXPH RI ZDWHU ZDV LQMHFWHG IURP WKH ILOOHG V\ULQJH LQWR WKH UHVSLURPHWHU 'HWHUPLQDWLRQV RI 32 ZHUH PDGH ZLWK D 5DGLRPHWHUp R[\JHQ HOHFWURGH FRQQHFWHG WR D 5DGLRPHWHUp 3+0 DFLGEDVH DQDO\]HU 7R EHJLQ D PHWDEROLF WULDO WKH PDLQ UHVHUYRLU DQG UHVSLURPHWHU ZHUH ILOOHG ZLWK ZDWHU DW WKH DFFOLPDWLRQ VDOLQLW\ )LVK ZHUH SODFHG LQWR WKH UHVSLURPHWHU K SULRU WR WKH EHJLQQLQJ RI D PHWDEROLF GHWHUPLQDWLRQ 7KLV DOORZHG ILVK VXIILFLHQW WLPH WR DGMXVW WR WKH H[SHULPHQWDO VHWXS 7KH UHVSLURPHWHU ZDV WKHQ VHDOHG DQG LPPHUVHG LQ WKH WKHUPRUHJXODWLRQ UHVHUYRLU DQG WKH VXEPHUVLEOH SXPS ZDV WXUQHG RQ DW D IORZ UDWH RI / PLQnr 7KH IROORZLQJ PRUQLQJ WKH SXPS ZDV WXUQHG RII WKH V\VWHP ZDV FORVHG DQG 32 PHDVXUHPHQWV EHJXQ 0HDVXUHPHQWV RI WKH UDWH RI UHGXFWLRQ LQ 32 ZHUH PDGH DW WR K LQWHUYDOV DQG FRQWLQXHG XQWLO ILVK KDG GHSOHWHG WKH R[\JHQ OHYHO WR DSSUR[LPDWHO\ PP +J JHQHUDOO\ WR Kf )ROORZLQJ WKH ILQDO 32 GHWHUPLQDWLRQ WKH V\VWHP ZDV

PAGE 53

7DEOH $FFOLPDWLRQ DQG ILQDO VDOLQLWLHV XVHG LQ VLPXODWHG WLGDO FKDQJH VWXG\ $FFOLPDWLRQ 6DOLQLW\ )LQDO 6DOLQLW\ r 2SSW SSW FRQWUROf SSW SSW SSW FRQWUROf SSW SSW SSW SSW FRQWUROf SSW SSW 2SSW SSW SSW FRQWUROf SSW SSW SSW SSW SSW SSW FRQWUROf SSW SSW SSW SSW FRQWUROf SSW SSW SSW SSW SSW FRQWUROf SSW SSW SSW FRQWUROf UHRSHQHG DQG WKH VXEPHUVLEOH SXPS WXUQHG RQ $W WKLV WLPH HLWKHU ZHOO ZDWHU SSWf RU D VDOLQH VROXWLRQ RI YDULDEOH FRQFHQWUDWLRQ ZDV SODFHG LQWR WKH VDOLQLW\ UHVHUYRLU 7KH VDOW FRQFHQWUDWLRQ LQ WKLV UHVHUYRLU ZDV GHWHUPLQHG VR WKDW LWV DGGLWLRQ WR WKH PDLQ UHVHUYRLU ZRXOG FKDQJH WKH VDOLQLW\ RI WKH PDLQ UHVHUYRLU DQG UHVSLURPHWHU WR WKH GHVLUHG ILQDO VDOLQLW\

PAGE 54

)LJXUH 6FKHPDWLF GLDJUDP RI UHVSLURPHWU\ DSSDUDWXV XVHG IRU URXWLQH PHWDEROLVP H[SHULPHQWV 6HH WH[W IRU GHWDLOHG GHVFULSWLRQ RI V\VWHP

PAGE 56

7KH YROXPHV RI ERWK UHVHUYRLUV DQG WKH UDWH RI SXPSLQJ E\ WKH SHULVWDOWLF SXPS ZHUH DGMXVWHG VR WKDW WKH VDOLQLW\ ZRXOG FKDQJH DW D XQLIRUP UDWH RYHU D K SHULRG 7KH SHULVWDOWLF SXPS ZDV SOXJJHG LQWR DQ DXWRPDWLF WLPHU VR WKDW VDOLQLW\ FKDQJHV WRRN SODFH EHWZHHQ K WR K 2Q WKH VHFRQG PRUQLQJ WKH V\VWHP ZDV UHFORVHG DQG 32 PHDVXUHPHQWV ZHUH PDGH DW WKH QHZ ILQDOf VDOLQLW\ 32 PHDVXUHPHQWV DW WKH ILQDO VDOLQLW\ ZHUH WKXV PDGH K IROORZLQJ FRPSOHWLRQ RI WKH VDOLQLW\ IOXFWXDWLRQ D SHULRG VXIILFLHQWO\ ORQJ WR EH SDVW WKH VWUHVVIXO WUDQVLWLRQ SHULRG 9RQ 2HUW]HQ f )ROORZLQJ WKH ILQDO 3 GHWHUPLQDWLRQ WKH ILVK ZDV UHPRYHG IURP WKH UHVSLURPHWHU GDPSGULHG DQG ZHLJKHG WR WKH QHDUHVW J $OO PHWDEROLF GHWHUPLQDWLRQV ZHUH PDGH EHWZHHQ K DQG K DQG ILVK ZHUH QRW UHn XVHG LQ RWKHU PHWDEROLF WULDOV )LVK ZHUH WKXV KHOG ZLWKLQ WKH UHVSLURPHWHU IRU D WRWDO RI WR K LQFOXGLQJ DGMXVWPHQW SHULRGf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f RI DOO LQGLYLGXDOV XVHG LQ WKLV VWXG\ $GMXVWHG 505 YDOXHV ZHUH FDOFXODWHG DV SUHYLRXVO\ GHVFULEHG $ UHSHDWHG PHDVXUHV DQDO\VLV RI YDULDQFH ZDV XVHG WR FRPSDUH DGMXVWHG 505nV EHWZHHQ DFFOLPDWLRQ DQG ILQDO VDOLQLWLHV 6WDWLVWLFDO DQDO\VHV IROORZ SURFHGXUHV RXWOLQHG LQ :LQHU HWDO f DQG 6RNDO DQG 5RKOI f $OO DQDO\VHV RI YDULDQFH ZHUH RQH ZD\ WHVWV XVLQJ WKH 7XNH\.UDPFU SRVW KRF FRPSDULVRQ S f

PAGE 57

5HVXOWV &RPSDULVRQ RI 505nV DW DFFOLPDWLRQ DQG ILQDO VDOLQLWLHV UHYHDOHG VRPH LQWHUHVWLQJ SDWWHUQV 7DEOH f 7KHUH ZHUH QR VLJQLILFDQW GLIIHUHQFHV S f LQ 505 EHWZHHQ DFFOLPDWLRQ DQG ILQDO VDOLQLWLHV LQ DQ\ FRQWURO WULDOV Q HDFKf 6LPLODUO\ ZKHQ ERWK DFFOLPDWLRQ DQG ILQDO VDOLQLWLHV SSW SSW SSW SSW DQG SSWf ZHUH LQ D UDQJH WKDW LV W\SLFDOO\ HQFRXQWHUHG E\ & YDULHJDWXV WKHUH ZDV D VLJQLILFDQW FKDQJH LQ 505 LQ RQO\ RQH WULDO 7\SLFDO VDOLQLWLHV LQ WKH ZLOG UDQJH IURP SSW WKURXJK SSW ZLWK K\SHUVDOLQH FRQGLWLRQV SSWf RFFXUULQJ PXFK PRUH IUHTXHQWO\ WKDQ VDOLQLWLHV QHDU SSW SHUV REVf :KHQ WKH DFFOLPDWLRQ VDOLQLW\ LV KLJK SSW DQG SSWf DOO JURXSV H[KLELWHG VLJQLILFDQW HOHYDWLRQ RI 505 LQ WKH ORZHU PRUH W\SLFDO ILQDO VDOLQLW\ 7KH VDPH JHQHUDO SDWWHUQ ZDV VHHQ ZKHQ WKH ILQDO VDOLQLW\ ZDV KLJK ZKHUH ILVK LQ WZR RI WKUHH JURXSV KDG GHSUHVVHG 505 DW WKH KLJKHVW VDOLQLWLHV 505 ZDV GHSUHVVHG ZKHQ HLWKHU WKH DFFOLPDWLRQ RU ILQDO VDOLQLW\ ZDV SSW 7KH GLUHFWLRQ RI VDOLQLW\ FKDQJH VWURQJO\ LQIOXHQFHG WKH PHWDEROLF UHVSRQVH :KHQ VDOLQLW\ ZDV LQFUHDVHG RYHU WKH FRXUVH RI WKH WULDO )LJXUH f ILVK ZHUH RQO\ DIIHFWHG PHWDEROLFDOO\ DW WKH YHU\ KLJKHVW DQG ORZHVW VDOLQLWLHV ZKHUH PHWDEROLVP ZDV GHSUHVVHG :KHQ VDOLQLW\ ZDV GHFUHDVHG RYHU WKH FRXUVH RI WKH WULDO )LJXUH f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

PAGE 58

7DEOH 0HDQ URXWLQH PHWDEROLVP PJ &K KnOf EHIRUH DFFOLPDWLRQ VDOLQLW\f DQG IROORZLQJ ILQDO VDOLQLW\f D VLPXODWHG WLGDO FKDQJH 9DOXHV DUH JLYHQ DV PHDQV s VH *URXSV H[KLELWLQJ D VLJQLILFDQW FKDQJH LQ PHWDEROLVP DUH LQGLFDWHG ZLWK DQ DVWHULVN $FFOLPDWLRQ 6DOLQLW\ 332 $FFOLPDWLRQ 505 PJ KnOf )LQDO 6DOLQLW\ 334 )LQDO 505 PJ &! KnOf 3 YDOXH 5HVSRQVH 4SSW s SSW Q f s r ,QFUHDVH SSW s SSW Q f s 1R &KDQJH s SSW Q f s 1R &KDQJH SSW s SSW Q f s 1R &KDQJH SSW s SSW Q f s r 'HFUHDVH s SSW Q f s r 'HFUHDVH s SSW Q f s 1R &KDQJH s SSW Q f s r 'HFUHDVH SSW s SSW Q f s 1R &KDQJH s SSW Q f s 1R &KDQJH s SSW Q f s 1R &KDQJH SSW s SSW Q f s 1R &KDQJH s SSW Q f s r 'HFUHDVH SSW s SSW Q f s r ,QFUHDVH s SSW Q f s r ,QFUHDVH SSW s SSW Q f s r ,QFUHDVH ZDV RQO\ ZKHQ WKH DFFOLPDWLRQ RU ILQDO VDOLQLWLHV ZHUH RXWVLGH WKH UDQJH QRUPDOO\ HQFRXQWHUHG LF SSW SSW DQG SSWf WKDW FKDQJHV LQ PHWDEROLVP ZHUH IRXQG ,Q WKHVH FDVHV PHWDEROLVP ZDV GHSUHVVHG DW HDFK H[WUHPH VDOLQLW\ UHODWLYH WR WKH PHDVXUHPHQWV PDGH DW WKH PRUH W\SLFDO VDOLQLW\

PAGE 59

)LJXUH 5HVXOWV RI PHWDEROLF WULDOV ZKHUH VDOLQLW\ ZDV LQFUHDVHG RYHU WKH FRXUVH RI WKH WULDO %DUV UHSUHVHQW JURXSV OLVWHG LQ 7DEOH IRU ZKLFK ILQDO VDOLQLW\ ZDV JUHDWHU WKDQ LQLWLDO VDOLQLW\ 7KH KHLJKW RI HDFK EDU VLJQLILHV WKH PDJQLWXGH RI WKH VDOLQLW\ FKDQJH IRU HDFK PHWDEROLF WULDO DQG WKH DVWHULVN LQGLFDWHV DW ZKLFK RI WKH VDOLQLWLHV IRU HDFK PHWDEROLF WULDOf WKH URXWLQH PHWDEROLF UDWH 505f ZDV KLJKHVW 7KH [ D[LV KDV QR VFDOH DQG VHUYHV RQO\ WR YLVXDOO\ VHSDUDWH JURXSV

PAGE 60

6DOLQLW\ SSWf 505nV 1RW 6LJQLILFDQWO\ 'LIIHUHQW S f 505nV 6LJQLILFDQWO\ 'LIIHUHQW S f r r r 6DOLQLW\ ,QFUHDVLQJ

PAGE 61

)LJXUH 5HVXOWV RI PHWDEROLF WULDOV ZKHUH VDOLQLW\ ZDV GHFUHDVHG RYHU WKH FRXUVH RI WKH WULDO %DUV UHSUHVHQW JURXSV OLVWHG LQ 7DEOH IRU ZKLFK ILQDO VDOLQLW\ ZDV OHVV WKDQ LQLWLDO VDOLQLW\ 7KH KHLJKW RI HDFK EDU VLJQLILHV WKH PDJQLWXGH RI WKH VDOLQLW\ FKDQJH IRU HDFK PHWDEROLF WULDO DQG WKH DVWHULVN LQGLFDWHV DW ZKLFK RI WKH VDOLQLWLHV IRU HDFK PHWDEROLF WULDOf WKH URXWLQH PHWDEROLF UDWH 505f ZDV KLJKHVW 7KH [ D[LV KDV QR VFDOH DQG VHUYHV RQO\ WR YLVXDOO\ VHSDUDWH JURXSV

PAGE 62

6DOLQLW\ SSWf 505nV 1RW 6LJQLILFDQWO\ 'LIIHUHQW S f 505nV 6LJQLILFDQWO\ 'LIIHUHQW S f r r r 6DOLQLW\ 'HFUHDVLQJ

PAGE 63

7KHVH UHVXOWV DOORZ IRU OLPLWHG FRPSDULVRQ EHFDXVH VLPLODU VWXGLHV KDYH H[DPLQHG WKH LQIOXHQFH RI VDOLQLW\ RYHU D PXFK QDUURZHU UDQJH WKDQ H[DPLQHG LQ WKLV VWXG\ 7KH EHVW FRPSDULVRQ LV ZLWK D VWXG\ E\ :DNHPDQ DQG :RKOVFKODJ f RQ WKH UHG GUXP 6FLDHQRSV RFHOODWXV ,Q WKHLU VWXG\ WHVW DQLPDOV ZHUH DFFOLPDWHG WR D VHULHV RI VDOLQLWLHV EHWZHHQ SSW DQG SSW DQG WKHQ DEUXSWO\ WUDQVIHUUHG WR VDOLQLWLHV HLWKHU SSW KLJKHU RU ORZHU WKDQ WKH DFFOLPDWLRQ VDOLQLW\ $OWKRXJK :DNHPDQ DQG :RKOVFKODJ f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f SSW YV WR SSW LQ WKH SUHVHQW VWXG\f +RZHYHU WKHVH UHVXOWV FRXOG DOVR EH DWWULEXWDEOH WR GLIIHUHQW VL]HV DQG DFWLYLW\ SDWWHUQV RI H[SHULPHQWDO ILVK EHWZHHQ WKH WZR VWXGLHV ,Q D VLPLODU VWXG\ 6KXVKPLQ f VWXGLHG WKH G\QDPLFV RI R[\JHQ FRQVXPSWLRQ LQ MXYHQLOH JROGHQ PXOOHW /L]D DXUDWD IROORZLQJ DEUXSW WUDQVIHUV IURP SSW WR D VHULHV RI VDOLQLWLHV UDQJLQJ IURP SSW WR SSW $OO VDOLQLW\ FKDQJHV UHVXOWHG LQ LQFUHDVHG PHWDEROLF UDWHV IRU RQH WR WKUHH GD\V IROORZLQJ WKH WUDQVIHU ZLWK PHWDEROLVP JHQHUDOO\ UHWXUQLQJ WR QHDU QRUPDO OHYHOV ZLWKLQ WKUHH WR IRXU GD\V ,W LV QRW VXUSULVLQJ WKDW VDOLQLW\ FKDQJHV DUF PRUH VWUHVVIXO LQ WKH JROGHQ PXOOHW VLQFH WKH VSHFLHV KDV ORZHU VDOLQLW\ WROHUDQFH DQG RVPRUHJXODWRU\ DELOLW\ WKDQ & YDULHJDWXV %DUWRQ DQG %DUWRQ f H[DPLQHG PHWDEROLVP LQ MXYHQLOH Jf & YDULHJDWXV FROOHFWHG IURP LQODQG VDOLQH ODNHV RI 6DQ 6DOYDGRU ,VODQG 7KH\ PHDVXUHG PHWDEROLF UDWHV RI

PAGE 64

ILVK DW SSW DQG SSW IROORZLQJ DEUXSW WUDQVIHU IURP DQ DFFOLPDWLRQ VDOLQLW\ RI SSW 0HWDEROLF UDWHV LQ ILVK PHDVXUHG DW SSW ZHUH VLJQLILFDQWO\ KLJKHU WKDQ WKRVH PDGH DW WKH DFFOLPDWLRQ VDOLQLW\ 7KHVH UHVXOWV DUH VRPHZKDW GLIIHUHQW IURP WKRVH REWDLQHG LQ WKH SUHVHQW VWXG\ +RZHYHU PHDVXUHPHQWV E\ %DUWRQ DQG %DUWRQ f ZHUH PDGH RQO\ K IROORZLQJ DEUXSW WUDQVIHU WR WKH UHGXFHG VDOLQLW\ ZKHUHDV PHDVXUHPHQWV LQ WKH SUHVHQW VWXG\ ZHUH PDGH K IROORZLQJ D JUDGXDO FKDQJH LQ VDOLQLW\ )LVK LQ WKLV VWXG\ WKXV H[SHULHQFHG D OHVV VWUHVVIXO WUDQVLWLRQ WR WKH QHZ VDOLQLW\ $Q RQWRJHQHWLF HIIHFW PD\ DOVR DFFRXQW IRU WKH REVHUYHG GLIIHUHQFHV DV PDQ\ LQYHVWLJDWLRQV KDYH GHPRQVWUDWHG WKDW DGXOWV DQG MXYHQLOHV RI WKH VDPH VSHFLHV PD\ GLIIHU JUHDWO\ LQ SK\VLRORJLFDO FRPSHWHQFH HJ .LQQH 0DUWLQ 2LNDZD HW DO f 2WKHU UHOHYDQW FRPSDULVRQV FDQ EH PDGH ZLWK GDWD RQ MXYHQLOH FURDNHU 0LFURSRJRQLDV XQGXODWXV DQG MXYHQLOH VSRW /HLRVWRPXV [DQWKXUXV 0RVHU DQG *HUU\ 0RVHU DQG 0LOOHU f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nO YV RU SSW KfOf DQG & YDULHJDWXV OLNHO\ H[SHULHQFHV JUHDWHU IOXFWXDWLRQV LQ VDOLQLW\ WKDQ GR VSRW RU FURDNHU $FFOLPDWLRQ VWDWH LV WKH PRVW LPSRUWDQW PHDVXUHG IDFWRU LQIOXHQFLQJ WKH PHWDEROLF UHVSRQVH RI & YDULHJDWXV WR VLPXODWHG WLGDO FKDQJHV LQ VDOLQLW\ +RZHYHU GLUHFWLRQ RI WKH VDOLQLW\ FKDQJH DOVR LQIOXHQFHG PHWDEROLVP LQ & YDULHJDWXV /LNH WKH UHVXOWV REWDLQHG E\ 0RVHU DQG 0LOOHU f RQ MXYHQLOH / [DQWKXUXV LW DSSHDUV WKDW & YDULHJDWXV DGMXVWV WR LQFUHDVLQJ VDOLQLW\ PRUH HIIHFWLYHO\ WKDQ WR GHFUHDVLQJ VDOLQLW\ DV HYLGHQFHG E\ WKH

PAGE 65

UHVSRQVHV WR VDOLQLW\ FKDQJHV EHWZHHQ SSW DQG SSW DQG EHWZHHQ SSW DQG SSW 7KXV ZKHQ VDOLQLW\ LV LQFUHDVHG RYHU WKH FRXUVH RI WKH WULDO ILVK DUH RQO\ DIIHFWHG PHWDEROLFDOO\ DW WKH YHU\n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

PAGE 66

&+$37(5 ,1)/8(1&( 2) $ )/8&78$7,1* 6$/,1,7< 5(*,0( 21 26025(*8/$7,21 ,1 &<35,12'21 9$5,(*$786 ,QWURGXFWLRQ )HZ DUHDV LQ WKH ILHOG RI ILVK SK\VLRORJ\ KDYH UHFHLYHG DV PXFK DWWHQWLRQ DV WKH VWXG\ RI RVPRUHJXODWLRQ 7KH EDVLF SDWWHUQV RI RVPRUHJXODWLRQ DUH ZHOO XQGHUVWRRG DQG DUH UHYLHZHG H[WHQVLYHO\ E\ (YDQV f .DPDN\ f 9HQWUHOOD HW DO f (YDQV f 0F&RUPLFN f DQG :RRG DQG 0DUVKDOO f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f WR VHDZDWHU SSWf 6XFK VSHFLHV WHQG WR EH VPDOO DQG RI OLWWOH RU QR HFRQRPLF LPSRUWDQFH ZLWK D QRWDEOH H[FHSWLRQ EHLQJ WKH PLONILVK &KDQRV FKDQRV )HUUDULV HW DO 6ZDQVRQ f +RZHYHU WR XQGHUVWDQG RVPRUHJXODWLRQ SDWWHUQV LQ ILVKHV VWXGLHV QHHG WR HQFRPSDVV WKH UDQJH RI VDOLQLWLHV HQFRXQWHUHG E\ VSHFLHV LQ QDWXUH 2QH WRSLF WKDW KDV UHFHLYHG OLWWOH DWWHQWLRQ FRQFHUQV WKH HIIHFW RI IOXFWXDWLQJ VDOLQLWLHV RQ RVPRUHJXODWLRQ LQ WFOFRVWV 6DOLQLW\ LV D OLPLWLQJ IDFWRU LQ FHUWDLQ HVWXDULQH DQG VDOW PDUVK KDELWDWV DQG PD\ ZLGHO\ YDU\ GDLO\ DQG VHDVRQDOO\ 'HVSLWH WKLV PRVW LQYHVWLJDWLRQV RI RVPRUHJXODWLRQ LQ ILVKHV LQYROYH VWXGLHV DW FRQVWDQW VDOLQLWLHV :KLOH VXFK VWXGLHV SURYLGH

PAGE 67

LPSRUWDQW LQIRUPDWLRQ RQ RYHUDOO RVPRUHJXODWRU\ SDWWHUQV WKH\ SURYLGH OHVV LQIRUPDWLRQ DERXW PRUH HFRORJLFDOO\ UHOHYDQW UHVSRQVHV 7KH DLP RI WKH SUHVHQW LQYHVWLJDWLRQ ZDV WZRIROG )LUVW H[DPLQHG WKH DELOLW\ RI LQGLYLGXDOV RI WKH HXU\KDOLQF WHOHRVW &\SULQRGRQ YDULHJDWXV WR UHJXODWH SODVPD RVPRODOLW\ XQGHU WKH LQIOXHQFH RI D F\FOLQJ VDOLQLW\ UHJLPH 6HFRQG H[DPLQHG D K\SRWKHVLV SURSRVHG E\ *RROLVK DQG %XUWRQ f LQ D VWXG\ LQYROYLQJ WKH LQWHUWLGDO FRSHSRG 7LJULRSXV FDOLIRUQLFXV *RROLVK DQG %XUWRQ f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f 1DWLRQDO %LRORJLFDO 6HUY LFH *DLQHVYLOOH )ORULGD )LVK ZHUH KHOG RYHUQLJKW LQ WKH FRROHUV XVHG IRU WUDQVSRUWDWLRQ ZLWK DHUDWLRQf EHIRUH WUDQVIHU WR D P GLDPHWHU ILEHUJODVV KROGLQJ WDQN FRQWDLQLQJ ZDWHU DW SSW )LVK ZHUH WUHDWHG SURSK\ODFWLFDOO\ IRU GD\V LQ D PJ /nO VROXWLRQ RI $FULIODYLQFp )ROORZLQJ WUHDWPHQW DOO ILVK ZHUH WUDQVIHUUHG WR H[SHULPHQWDO DTXDULD SSW VDOLQLW\ P2VP NJnOf ORFDWHG ZLWKLQ D FRQVWDQW HQYLURQPHQW URRP PDLQWDLQHG DW s r& DQG RQ D

PAGE 68

K OLJKWGDUN F\FOH %RWK KROGLQJ DQG H[SHULPHQWDO DTXDULD ZHUH HTXLSSHG ZLWK VSRQJH ILOWHUV SURYLGLQJ FRQWLQXRXV DHUDWLRQ DQG ILVK ZHUH IHG IODNH IRRG RQFH HDFK GD\ ([SHULPHQWDO DTXDULD ZHUH XVHG WR VXEMHFW ILVK WR D F\FOLQJ VDOLQLW\ UHJLPH 6HYHQ VDOLQLW\ WULDOV ZHUH SHUIRUPHG 7DEOH f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f $ VXEVDPSOH RI ILVK ZDV UHPRYHG IRU WHVWLQJ IURP HDFK H[SHULPHQWDO DTXDULXP MXVW SULRU WR WKH ILUVW FKDQJH LQ VDOLQLW\ F\FOH f DQG DW WKH HQG RI F\FOHV GD\ f GD\ f DQG GD\ f $OO VXEVDPSOHV ZHUH WKXV WDNHQ IROORZLQJ D K SHULRG LQ EDQN DTXDULD SSWf )ROORZLQJ FRPSOHWLRQ RI WKH WK F\FOH ILVK UHPDLQLQJ LQ DOO GHFUHDVLQJ VDOLQLW\ JURXSV JURXSV 'L (E DQG 'f ZHUH WUDQVIHUUHG WR DTXDULD DW SSW DQG ILVK UHPDLQLQJ LQ DOO LQFUHDVLQJ VDOLQLW\ JURXSV JURXSV ,M ) DQG ,f ZHUH WUDQVIHUUHG WR DTXDULD DW SSW )LVK LQ WKH DTXDULD PDLQWDLQHG DW SSW ZHUH VSOLW LQWR WKUHH JURXSV DW WKDW WLPH RQH WKLUG RI WKH JURXS ZDV WUDQVIHUUHG WR SSW JURXS &Gf RQH WKLUG WUDQVIHUUHG WR SSW JURXS &f DQG WKH ILQDO WKLUG WUDQVIHUUHG WR SSW JURXS 4f )LVK UHPDLQHG LQ WKHVH VDOLQLWLHV IRU K DQG ZHUH WKHQ UHPRYHG IRU WHVWLQJ GD\ f 6DOLQLWLHV ZHUH FKHFNHG GDLO\ ZLWK D /HLFDp WHPSHUDWXUH FRPSHQVDWHG UHIUDFWRPHWHU DQG DGMXVWHG DV QHFHVVDU\ +HPDWRFULW +FWf DQG SODVPD RVPRODOLW\ ZHUH GHWHUPLQHG DW HDFK VDPSOLQJ LQWHUYDO )LVK ZHUH ILUVW FDUHIXOO\ QHWWHG IURP WKHLU H[SHULPHQWDO DTXDULD DQG EORWWHG GU\ %ORRG ZDV WDNHQ E\ VWHUQDO FDUGLDF SXQFWXUH XVLQJ KHSDULQL]HG PLFURKFPDWRFULW WXEHV GUDZQ WR D ILQH SRLQW DQG ILVK ZHUH ZHLJKHG DQG VWDQGDUG OHQJWK GHWHUPLQHG 7KH WXEHV ZHUH WKHQ

PAGE 69

7DEOH 6DOLQLW\ WULDOV XVHG LQ F\FOLFDO VDOLQLW\ VWXG\ 7KH JURXS PDLQWDLQHG DW SSW ZDV VSOLW LQWR WKUHH JURXSV IROORZLQJ F\FOH GD\ f JURXSV &T & DQG 4 VHH WH[W IRU GHWDLOVf *URXS 'LUHFWLRQ RI 6DOLQLW\ &KDQJH 6DOLQLW\ LQ %DQN 332 6DOLQLW\ LQ %DQN 332 'L 'HFUHDVLQJ 6DOLQLW\ Q f G 'HFUHDVLQJ 6DOLQLW\ Q f G 'HFUHDVLQJ 6DOLQLW\ Q f & &G DQG 4 1R &KDQJH Q f ,W ,QFUHDVLQJ 6DOLQLW\ Q f O ,QFUHDVLQJ 6DOLQLW\ Q f ,QFUHDVLQJ 6DOLQLW\ Q f FHQWULIXJHG LQ D PLFURKHPDWRFULW FHQWULIXJH IRU PLQXWHV WR VHSDUDWH SODVPD IURP FHOOV +HPDWRFULW ZDV UHDG XVLQJ D PLFURFDSLOODU\ UHDGHU EHIRUH SODVPD ZDV LVRODWHG IURP IRUPHG HOHPHQWV E\ VFRULQJ WKH WXEH ZLWK D ILOH DQG UHWDLQLQJ RQO\ WKH SRUWLRQ FRQWDLQLQJ SODVPD 3ODVPD RVPRODOLW\ P2VP NJff ZDV GHWHUPLQHG RQ SL VDPSOHV XVLQJ D :HVFRUp YDSRU SUHVVXUH RVPRPHWHU )LVK ZHUH XVHG ZLWKRXW UHJDUG WR VH[ DQG DOO EORRG VDPSOHV ZHUH WDNHQ EHWZHHQ K DQG K 6WDWLVWLFDO SURFHGXUHV IROORZHG :LQHU HW DO f DQG 6RNDO DQG 5RKOI f $OO VWDWLVWLFDO DQDO\VHV ZHUH RQH ZD\ WHVWV XVLQJ WKH 7XNH\ .UDPHU SRVW KRF FRPSDULVRQ S f 5HVXOWV $OO ILVK HQWHUHG LQWR WKH H[SHULPHQWDO SURFHGXUH VXUYLYHG WKH HQWLUH GXUDWLRQ RI WKH H[SHULPHQW )LVK LQ DOO JURXSV DWH QRUPDOO\ WKURXJKRXW WKH FRXUVH RI WKH H[SHULPHQW DQG QR GLVFHUQLEOH FKDQJHV LQ EHKDYLRU ZHUH REVHUYHG GXULQJ WKH H[SHULPHQWDO SURFHGXUH IRU DQ\

PAGE 70

JURXS )LVK DSSHDUHG WR KDYH QR GLIILFXOW\ LQ WROHUDWLQJ WKH LPSRVHG VDOLQLW\ IOXFWXDWLRQV HYHQ LQ JURXSV 'L DQG ZKLFK ZHUH VXEMHFWHG WR GDLO\ FKDQJHV LQ VDOLQLW\ RI SSW DQG SSW UHVSHFWLYHO\ 7KH LQIOXHQFH RI ERG\ PDVV RQ ERWK +FW DQG SODVPD RVPRODOLW\ ZDV HYDOXDWHG XVLQJ DQ DQDO\VLV RI FRYDULDQFH DQG IRU QR DQDO\VLV ZDV ERG\ PDVV D VLJQLILFDQW FRYDULDWH 6L]HV RI ILVK XVHG LQ WKLV VWXG\ UDQJHG IURP J WR J PP WR PP VWDQGDUG OHQJWK PHDQ s PPf ZLWK D PHDQ PDVV RI s J 6XEVDPSOHV RI JURXSV ZHUH QRW VLJQLILFDQWO\ GLIIHUHQW LQ PDVV +HPDWRFULW DQG SODVPD RVPRODOLW\ YDOXHV ZHUH FRPSDUHG ERWK EHWZHHQ VDOLQLW\ WULDOV IRU HDFK WLPH SHULRG VDPSOHG LH DOO JURXSV ZHUH FRPSDUHG WR RQH DQRWKHU RQ GD\V DQG f DQG ZLWKLQ D VLQJOH WULDO RYHU WKH WLPH FRXUVH RI WKH H[SHULPHQW *URXSV H[SHULHQFLQJ LQFUHDVHV LQ VDOLQLW\ JURXSV W\ ) DQG 4f ZHUH FRPSDUHG VHSDUDWHO\ IURP JURXSV H[SHULHQFLQJ GHFUHDVHV LQ VDOLQLW\ JURXSV 'L &W\ DQG &Gf ZLWK DOO FRPSDUHG WR WKH JURXS PDLQWDLQHG DW D FRQVWDQW VDOLQLW\ JURXS &f 'DWD IURP GD\V WKURXJK DUF GLVFXVVHG VHSDUDWHO\ IURP GD\ GDWD DV IROORZV 'D\ WKURXJK +HPDWRFULW YDOXHV VKRZHG QR REYLRXV WUHQGV HLWKHU EHWZHHQ VDOLQLW\ WULDOV RU ZLWKLQ DQ\ VLQJOH WULDO RYHU WKH WLPH FRXUVH RI WKH H[SHULPHQW 7DEOH f 7KHUH ZHUH QR VWDWLVWLFDOO\ VLJQLILFDQW GLIIHUHQFHV S f ZLWKLQ DQ\ JURXS EHWZHHQ GD\V DQG f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f )RU JURXSV & DQG ,M WKLV ZDV VWDWLVWLFDOO\ VLJQLILFDQW ZLWK

PAGE 71

7DEOH 5HVXOWV RI VDOLQLW\ IOXFWXDWLRQV H[SHULPHQW 9DOXHV LQ WKH WRS URZ RI HDFK FHOO UHSUHVHQW KHPDWRFULW PHDVXUHPHQWV b HU\WKURF\WHVf YDOXHV LQ ERWWRP URZ RI HDFK FHOO UHSUHVHQW SODVPD RVPRODOLW\ PHDVXUHPHQWV P2VP NJrE 6DPSOH VL]HV DUH Q IRU HDFK FHOO $OO YDOXHV DUH H[SUHVVHG DV PHDQV s VH 6HH WH[W IRU H[SODQDWLRQ RI JURXS DEEUHYLDWLRQV *5283 '$< G G &G & &O K E E s s s f§ s f§ s s s s s s s s s s s s s f§ s f§ s s s s s s s s s s s s s f§ s f§ s s s s s s s s s s s s s f§ s f§ s s s s s s s s s s s s s s s s s s s s s s s s s s s s

PAGE 72

)LJXUH 0HDQ SODVPD RVPRODOLW\ YDOXHV PHDVXUHG IRU JURXSV H[SHULHQFLQJ GHFUHDVHV LQ VDOLQLW\ GXULQJ WKH FRXUVH RI WKH H[SHULPHQW *URXS GHVLJQDWLRQV DUH DV IROORZV 'M VDOLQLW\ IOXFWXDWHG EHWZHHQ SSW DQG SSW VDOLQLW\ IOXFWXDWHG EHWZHHQ SSW DQG SSW VDOLQLW\ IOXFWXDWHG EHWZHHQ SSW DQG SSW &R VDOLQLW\ FRQVWDQW DW SSW IRU GD\V GHFUHDVHG WR SSW IROORZLQJ VXEVDPSOH RQ GD\ & VDOLQLW\ FRQVWDQW DW SSW 6DPSOH VL]HV DUH Q IRU HDFK JURXS 6HH WH[W IRU GHWDLOV RI H[SHULPHQWDO SURFHGXUH

PAGE 73

F

PAGE 74

)LJXUH 0HDQ SODVPD RVPRODOLW\ YDOXHV PHDVXUHG IRU JURXSV H[SHULHQFLQJ LQFUHDVHV LQ VDOLQLW\ GXULQJ WKH FRXUVH RI WKH H[SHULPHQW *URXS GHVLJQDWLRQV DUH DV IROORZV M VDOLQLW\ IOXFWXDWHG EHWZHHQ SSW DQG SSW VDOLQLW\ IOXFWXDWHG EHWZHHQ SSW DQG SSW VDOLQLW\ IOXFWXDWHG EHWZHHQ SSW DQG SSW 4 VDOLQLW\ FRQVWDQW DW SSW IRU GD\V LQFUHDVHG WR SSW IROORZLQJ VXEVDPSOH RQ GD\ & VDOLQLW\ FRQVWDQW DW SSW 6DPSOH VL]HV DUH Q IRU HDFK JURXS 6HH WH[W IRU GHWDLOV RI H[SHULPHQWDO SURFHGXUH

PAGE 75

3ODVPD 2VPRWLF &RQFHQWUDWLRQ P2VP OHJ WR WR B WR :

PAGE 76

WKH YDOXHV RQ GD\ VLJQLILFDQWO\ KLJKHU WKDQ YDOXHV IURP GD\ DQG GD\ S[f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f IROORZLQJ WKH ILQDO VDOLQLW\ IOXFWXDWLRQ +HPDWRFULW UHVXOWV VKRZHG QR FRQVLVWHQW WUHQGV RQ GD\ VDPSOHV 2QO\ RQH VLJQLILFDQW GLIIHUHQFH ZDV QRWHG ZLWK WKH +FW YDOXH IRU JURXS 4 EHLQJ VLJQLILFDQWO\ HOHYDWHG FRPSDUHG WR WKH YDOXH IRU JURXS & Sf $OO RWKHU FRPSDULVRQV VKRZHG QR VLJQLILFDQW GLIIHUHQFHV LQ +FW 7KHUH ZHUH VLJQLILFDQW GLIIHUHQFHV LQ SODVPD RVPRODOLW\ IRU GD\ PHDVXUHPHQWV +HUH JURXSV ZKLFK KDG H[SHULHQFHG JUHDWHU IOXFWXDWLRQV LQ VDOLQLW\ RYHU WKH FRXUVH RI WKH H[SHULPHQW VKRZHG D PXFK EHWWHU DELOLW\ WR RVPRUHJXODWH FRPSDUHG WR WKH FRQWURO JURXS RU JURXSV ZKLFK KDG XQGHUJRQH VPDOO VDOLQLW\ IOXFWXDWLRQV ,QFUHDVLQJ VDOLQLW\ VHHPHG WR KDYH D JUHDWHU LPSDFW RQ SODVPD RVPRODOLW\ WKDQ GLG GHFUHDVLQJ VDOLQLW\ )LJXUHV DQG f 3ODVPD RVPRODOLW\ LQ JURXSV 'M >! DQG DOO H[KLELWHG GHFOLQHV RQ GD\ UHODWLYH WR GD\ EXW WKHVH GLIIHUHQFHV ZHUH QRW VWDWLVWLFDOO\ VLJQLILFDQW 1R GLIIHUHQFHV LQ UHJXODWRU\ DELOLW\ DPRQJ WKHVH WKUHH JURXSV RU ZLWK WKH FRQWURO JURXS JURXS &f FRXOG EH

PAGE 77

GLVFHUQHG +RZHYHU JURXS &T ZKLFK KDG EHHQ PDLQWDLQHG DW SSW IRU WKH ILUVW GD\V RI WKH H[SHULPHQW GLG VKRZ D VLJQLILFDQW GHFUHDVH LQ SODVPD RVPRODOLW\ RQ GD\ UHODWLYH WR YDOXHV PHDVXUHG RQ GD\V WKURXJK )XUWKHUPRUH WKLV JURXS KDG ORZHU RVPRODOLW\ YDOXHV RQ GD\ WKDQ DOO RWKHU JURXSV DOWKRXJK WKLV GLIIHUHQFH ZDV RQO\ VWDWLVWLFDOO\ VLJQLILFDQW ZKHQ FRPSDUHG WR JURXSV & DQG 0RUH VWULNLQJ GLIIHUHQFHV H[LVWHG IRU JURXSV K DQG 4 ,Q DOO FDVHV SODVPD RVPRODOLW\ ZDV HOHYDWHG RQ GD\ UHODWLYH WR PHDVXUHPHQWV WDNHQ IURP WKH VDPH JURXS RQ GD\ RU HDUOLHU *URXS ZKLFK KDG H[SHULHQFHG GDLO\ IOXFWXDWLRQV LQ VDOLQLW\ RI SSW VKRZHG WKH VPDOOHVW LQFUHDVH LQ RVPRODOLW\ YDOXHV EHLQJ VLJQLILFDQWO\ KLJKHU WKDQ PHDVXUHPHQWV IURP GD\ RQO\ *URXS 8 VKRZHG D VLPLODU UHVXOW ZLWK WKH SODVPD RVPRODOLW\ YDOXH IURP GD\ VLJQLILFDQWO\ KLJKHU WKDQ YDOXHV IURP GD\ DQG GD\ RQO\ *URXSV ,L DQG &_ VKRZHG WKH ODUJHVW FKDQJHV LQ SODVPD RVPRODOLW\ RQ GD\ %RWK JURXSV KDG RVPRODOLW\ YDOXHV WKDW ZHUH VLJQLILFDQWO\ KLJKHU WKDQ DOO SUHYLRXV PHDVXUHPHQWV WDNHQ IURP WKHLU UHVSHFWLYH JURXSV 7KH YDOXH IRU JURXS ,L ZDV HOHYDWHG FRPSDUHG WR JURXSV DQG & DOWKRXJK WKLV LQFUHDVH ZDV VWDWLVWLFDOO\ VLJQLILFDQWO\ RQO\ ZKHQ FRPSDUHG WR JURXS & *URXS &_ ZDV VLJQLILFDQWO\ HOHYDWHG ZKHQ FRPSDUHG WR DOO RWKHU JURXSV LQFOXGLQJ JURXS ,Wf 'LVFXVVLRQ 7KH DELOLW\ WR DGMXVW UDSLGO\ WR DOWHUHG VDOLQLWLHV ZRXOG EH DQ REYLRXV DGYDQWDJH WR VDOW PDUVK RUJDQLVPV 3K\VLRORJLFDO UHVSRQVHV RI HXU\KDOLQH ILVKHV H[SRVHG WR UDSLG FKDQJHV LQ VDOLQLW\ FDQ EH JURXSHG LQWR WZR SKDVHV +ROPHV DQG 'RQDOGVRQ f DQ DGDSWLYH SHULRG DQG D UHJXODWRU\ SHULRG 'XULQJ WKH DGDSWLYH SHULRG SODVPD RVPRODOLW\ YDULHV JUDGXDOO\ UHWXUQLQJ WR YDOXHV DSSURDFKLQJ RULJLQDO OHYHOV ,Q WKH UHJXODWRU\ SHULRG SODVPD RVPRODOLW\ LV PRUH ILQHO\ FRQWUROOHG DV WKH ILVK DGMXVWV WR WKH DOWHUHG VDOLQLW\ DQG UHDFKHV LRQLF KRPHRVWDVLV )LVKHV ZKLFK UHDFK WKH UHJXODWRU\ SHULRG TXLFNO\ LH KDYH VKRUW DGDSWLYH SHULRGVf VKRXOG EH EHVW DEOH WR WROHUDWH DOWHUDWLRQV LQ DPELHQW VDOLQLW\ $OWKRXJK WKH

PAGE 78

OHQJWK RI WKH DGDSWLYH SHULRG ZDV QRW PHDVXUHG LQ WKH SUHVHQW VWXG\ UHVXOWV QHYHUWKHOHVV VXJJHVW WKDW LW LV UHODWLYHO\ VKRUW IRU & YDULHJDWXV 1RUGOLH f VKRZHG WKDW & YDULHJDWXV ZDV DQ H[FHOOHQW RVPRWLF UHJXODWRU RYHU D VDOLQLW\ UDQJH IURP SSW WR SSW 3ODVPD RVPRODOLW\ YDOXHV FKDQJHG RQO\ VOLJKWO\ RYHU WKLV UDQJH RI VDOLQLWLHV LQ KLV VWXG\ YDU\LQJ E\ RQO\ P2VP NJnO +RZHYHU ILVK LQ KLV VWXG\ ZHUH IXOO\ DFFOLPDWHG WR HDFK H[SHULPHQWDO VDOLQLW\ SULRU WR WHVWLQJ 7KH SUHVHQW VWXG\ LQGLFDWHV WKDW & YDULHJDWXV LV DQ H[FHOOHQW UHJXODWRU RI SODVPD RVPRODOLW\ HYHQ ZKHQ ILVKHV DUH H[SRVHG WR ODUJH GDLO\ IOXFWXDWLRQV LQ VDOLQLW\ $OWKRXJK ILVK LQ DOO JURXSV LQ WKH SUHVHQW VWXG\ UHJXODWHG DW VOLJKWO\ KLJKHU OHYHOV WKDQ VHHQ E\ 1RUGOLH f SODVPD RVPRODOLW\ YDOXHV YDULHG VLPLODUO\ ZLWK GLIIHUHQFHV RI OHVV WKDQ P2VP NJn IRU DOO JURXSV RQ GD\V WKURXJK 7KH WUDQVIHU SURFHVV LWVHOI HOLFLWHG PXFK RI WKLV YDULDWLRQ ZLWK VOLJKW LQFUHDVHV LQ SODVPD RVPRODOLW\ VHHQ RYHU WKH WLPH FRXUVH RI WKH H[SHULPHQW $ VLPLODU WUHQG ZDV VHHQ LQ D VWXG\ E\ :RR DQG :X f RQ WKH UHG JURXSHU (SLQHSKHOXV DNDDUD DQG WKH EODFN VHD EUHDP 0\ OLR PDFURFHSKDOXV 7KH LQIOXHQFH RI D VLQJOH DOWHUDWLRQ LQ VDOLQLW\ RQ RVPRUHJXODWRU\ DELOLW\ KDV EHHQ VWXGLHG IRU D QXPEHU RI ILVKHV ZLWK FKDQJHV LQ SODVPD RVPRODOLW\ REVHUYHG LQ WKH SUHVHQW VWXG\ RI VLPLODU PDJQLWXGH WR WKRVH VHHQ LQ RWKHU FXU\KDOLQH ILVKHV :DNHPDQ DQG :RKOVFKODJ (QJHO FW DO )HUUDULV HW DO 0DQFHUD HW DO
PAGE 79

7KHVH UHVXOWV VXJJHVW WKDW WKHUH DUH QR ODUJH PRYHPHQWV RI ZDWHU EHWZHHQ WKH EORRG DQG WLVVXHV DV D UHVXOW RI IOXFWXDWLRQV LQ VDOLQLW\ RYHU WKH UDQJH WHVWHG 7KH ODFN RI UHVSRQVH LQ KHPDWRFULW LQ WKLV VWXG\ PD\ DOVR LQGLFDWH WKDW ZLWKLQ WKH UDQJH RI VDOLQLWLHV VWXGLHG & YDULHJDWXV LV VXEMHFW WR QRUPDO RU WROHUDQFH SK\VLRORJLFDO SURFHVVHV DV DQ LQFUHDVH LQ KHPDWRFULW YDOXHV ZRXOG EH H[SHFWHG LI & YDULHJDWXV ZHUH H[SRVHG WR FRQGLWLRQV OHDGLQJ WR UHVLVWDQFH SURFHVVHV 6ZLIW f ,W KDV EHHQ K\SRWKHVL]HG WKDW ILVKHV RFFXUULQJ LQ D KDELWDW ZKHUH VDOLQLW\ RIWHQ IOXFWXDWHV PD\ EH DEOH WR UHVSRQG PRUH TXLFNO\ DQG FRPSOHWHO\ WR DOWHUDWLRQV LQ VDOLQLW\ DQG WKDW WKH OLPLWV RI WROHUDQFH DUH IDUWKHU DSDUW LI VDOLQLW\ IOXFWXDWHV SHULRGLFDOO\ *XQWHU 6SDDUJDUHQ -RKQVWRQ DQG &KHYHULH f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

PAGE 80

H[SHULPHQWDO JURXSV WKHVH GLIIHUHQFHV ZHUH QRW ODUJH HQRXJK LQ PDJQLWXGH WR FDXVH REVHUYDEOH GLVWUHVV LQ WKH H[SHULPHQWDO DQLPDOV 7KHVH UHVXOWV LQGLFDWH WKDW & YDULHJDWXV LV ZHOO DGMXVWHG IRU OLIH LQ YDULDEOH VDOLQLW\ FRQGLWLRQV ,W ZRXOG EH LQWHUHVWLQJ WR FRPSDUH UHVXOWV IURP WKH SUHVHQW H[SHULPHQWV ZLWK VWXGLHV H[DPLQLQJ WKH LQIOXHQFH RI IOXFWXDWLQJ VDOLQLW\ RQ RVPRUHJXODWLRQ LQ ILVKHV WKDW FDQ WROHUDWH FKDQJLQJ VDOLQLW\ EXW WKDW H[SHULHQFH LQIUHTXHQW VDOLQLW\ YDULDWLRQV LQ WKHLU QDWLYH KDELWDW

PAGE 81

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f )XUWKHUPRUH ILVKHV LQ VDOLQH ZDWHU ZLWK ORZ R[\JHQ WHQVLRQ PXVW EDODQFH PD[LPL]LQJ EUDQFKLDO R[\JHQ GLIIXVLRQ ZLWK JUHDWHU RVPRUHJXODWRU\ GHPDQGV GXH WR WKH DFFRPSDQ\LQJ LQFUHDVHV LQ LRQ DQG ZDWHU H[FKDQJH 3HUU\ DQG 0F'RQDOG f $GGLWLRQDOO\ WKH R[\JHQ FRQWHQW RI PDQ\ DTXDWLF KDELWDWV LV VXEMHFW WR ODUJH QDWXUDO IOXFWXDWLRQV VR R[\JHQ LV D SRWHQWLDOO\ OLPLWLQJ IDFWRU E\ LWVHOI 'HMRXUV *UDKDP f 7KLV LV HVSHFLDOO\ WUXH LQ VKDOORZ ZDWHUV ZKHUH FKURQLF RU SHULRGLF K\SR[LD PD\ EH D FRPPRQ SKHQRPHQRQ *UDKDP f 6DOW PDUVK KDELWDWV DUH RIWHQ H[SRVHG WR K\SR[LF FRQGLWLRQV 5HQDXG 7RXOPRQG f &\SULQRGRQ YDULHJDWXV LV DQ H[WUHPHO\ FRPSHWHQW HXU\KDOLQH WFOHRV/ 2YHU D UDQJH RI VDOLQLWLHV IURP IUHVKZDWHU SSWf WR SSW YHU\ VPDOO FKDQJHV LQ PHWDEROLVP RFFXU 1RUGOLF HW DO WKLV VWXG\ &KDSWHU f %RWK R[\JHQ FRQVXPSWLRQ DQG FULWLFDO R[\JHQ WHQVLRQ 3Ff DUF HVVHQWLDOO\ XQDIIHFWHG E\ FKDQJHV LQ VDOLQLW\ RYHU WKLV UDQJH 6DOLQLWLHV DERYH WKLV UDQJH FDXVH PHWDEROLF DGMXVWPHQWV ZLWK LQFUHDVHV LQ 3F DQG GHFUHDVHV LQ PHWDEROLVP REVHUYHG WKLV VWXG\ &KDSWHU f 7KH PHFKDQLVPV LQYROYHG LQ PDLQWDLQLQJ

PAGE 82

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f H[DPLQHG KHPRJORELQ FRQFHQWUDWLRQ HU\WKURF\WH FRXQW DQG SDFNHG FHOO YROXPH KHPDWRFULWf RYHU D ZLGH UDQJH RI VDOLQLWLHV IRU LQGLYLGXDOV RI & YDULHJDWXV WR HYDOXDWH WKH LQIOXHQFH RI VDOLQLW\ RQ EORRG R[\JHQ OHYHOV LQ WKLV H[WUHPHO\ FXU\KDOLQH VSHFLHV 0HWKRGV &ROOHFWLRQV RI ILVK WUDQVSRUWDWLRQ EDFN WR WKH ODERUDWRU\ DQG JHQHUDO ODE SURFHGXUHV ZHUH SHUIRUPHG DV GHVFULEHG SUHYLRXVO\ 8VLQJ WKH VDPH SURWRFRO GHVFULEHG HDUOLHU ILVK ZHUH VHTXHQWLDOO\ DFFOLPDWHG WR D VHULHV RI VDOLQLWLHV UDQJLQJ IURP SSW WR SSW WR P2VP NJfOf $W WKH HQG RI WKH DFFOLPDWLRQ SHULRG ILVK ZHUH VDFULILFHG WR GHWHUPLQH KHPRJORELQ FRQFHQWUDWLRQ >+E@f KHPDWRFULW +FWf DQG HU\WKURF\WH 5%&f FRXQW %ORRG 6DPSOLQJ )LVK ZHUH ILUVW FDUHIXOO\ QHWWHG IURP WKHLU H[SHULPHQWDO DTXDULD DQG EORWWHG GU\ %ORRG ZDV WDNHQ E\ VWHUQDO FDUGLDF SXQFWXUH XVLQJ IUHVKO\ KHSDULQL]HG PLFURKFPDWRFULW WXEHV GUDZQ WR D ILQH SRLQW %ORRG IURP HDFK ILVK ZDV FROOHFWHG LQ WZR PLFURKFPDWRFULW

PAGE 83

WXEHV 2QFH WKH ILUVW WXEH ZDV ILOOHG ZLWK D YROXPH SL WKH EORRG ZDV LPPHGLDWHO\ GLVSHQVHG LQWR D VPDOO FHUDPLF FUXFLEOH (SSHQGRUIp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f DQG 6RNDO DQG 5RKOI f $OO VWDWLVWLFDO DQDO\VHV ZHUH RQH ZD\ WHVWV XVLQJ WKH 7XNH\.UDPHU SRVW KRF FRPSDULVRQ S f +HPRJORELQ $QDO\VLV +HPRJORELQ FRQFHQWUDWLRQ ZDV PHDVXUHG VSHFWURSKRWRPHWULFDOO\ RQ SL VDPSOHV XVLQJ WKH F\DQRPHWKHPRJORELQ PHWKRG %URZQ f $V UHFRPPHQGHG E\ ,QQHV DQG :HOOV f WKH F\DQRPHWKHPRJORELQ VROXWLRQV ZHUH FHQWULIXJHG IRU PLQ DW J SULRU WR FRORULPHWULF GHWHUPLQDWLRQ WR UHPRYH HU\WKURF\WH GHEULV 7KLV PHWKRG JLYHV UHOLDEOH UHVXOWV ZKHQ XVHG ZLWK ILVK EORRG %OD[KDOO %OD[KDOO DQG 'DLVOH\ &REXUQ ,QQHV DQG :HOOV f (U\WKURF\WH &RXQW (U\WKURF\WHV ZHUH FRXQWHG LPPHGLDWHO\ IROORZLQJ GLOXWLRQ LQ WHVW WXEHV FRQWDLQLQJ 1DWW DQG +HUULFNV VROXWLRQ &DPSEHOO DQG 0XUUX f 7KLV VROXWLRQ DFWV DV ERWK VWDLQ DQG GLOXWHQW DQG LV URXWLQHO\ XVHG IRU FRXQWLQJ HU\WKURF\WHV RI ILVK &DPSEHOO DQG 0XUUX f $ GLOXWLRQ ZDV XVHG DQG FHOOV ZHUH FRXQWHG LQ DQ LPSURYHG 1FXEDXHU KFPDWRF\WRPFWHU IROORZLQJ SUHFDXWLRQV RXWOLQHG LQ %URZQ f

PAGE 84

+HPDWRFULW +HPDWRFULW ZDV PHDVXUHG WR GHWHUPLQH WKH SDFNHG FHOO YROXPH RI WKH HU\WKURF\WHV FRQWDLQHG LQ WKH EORRG ,PPHGLDWHO\ DIWHU ILOOLQJ WKH VHFRQG PLFURKHPDWRFULW WXEH RQH HQG ZDV VHDOHG DQG WKH WXEH SODFHG LQWR D PLFURKHPDWRFULW FHQWULIXJH 7XEHV ZHUH WKHQ FHQWULIXJHG IRU PLQ WR VHSDUDWH SODVPD IURP IRUPHG HOHPHQWV 7KH KHPDWRFULW ZDV UHDG XVLQJ D PLFURFDSLOODU\ UHDGHU DQG H[SUHVVHG DV SHUFHQW HU\WKURF\WH )URP WKH WHVW YDOXHV REWDLQHG PHDQ FRUSXVFXODU YROXPH 0&9f PHDQ FRUSXVFXODU KHPRJORELQ 0&+f DQG PHDQ FRUSXVFXODU KHPRJORELQ FRQFHQWUDWLRQ 0&+&f ZHUH FDOFXODWHG IRU HDFK ILVK DV IROORZV %URZQ f +FPRO 0&+B>W:-f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f 9DOXHV REWDLQHG DW SSW DQG SSW ZHUH VLJQLILFDQWO\ KLJKHU WKDQ DOO RWKHU VDOLQLWLHV (U\WKURF\WH FRXQW ZDV QH[W KLJKHVW DW SSW DQG SSW EHLQJ VLJQLILFDQWO\ HOHYDWHG FRPSDUHG WR YDOXHV LQ ILVK DFFOLPDWHG WR DQG SSW )LVK DFFOLPDWHG WR VDOLQLWLHV IURP SSW WKURXJK SSW H[KLELWHG QR VLJQLILFDQW GLIIHUHQFHV LQ HU\WKURF\WH FRXQW 0HDVXUHPHQWV RI KHPRJORELQ FRQFHQWUDWLRQ H[KLELWHG D VLPLODU SDWWHUQ DOWKRXJK IHZHU VLJQLILFDQW GLIIHUHQFHV ZHUH QRWHG )LJXUH f 0HDQ YDOXHV RI ILVKHV DFFOLPDWHG WR SSW ZHUH KLJKHVW DQG VLJQLILFDQWO\ GLIIHUHQW IURP ILVK DFFOLPDWHG WR VDOLQLWLHV IURP SSW

PAGE 85

7DEOH +HPDWRFULW +FWf KHPRJORELQ FRQFHQWUDWLRQ >+E@f HU\WKURF\WH FRXQW 5%&f PHDQ FRUSXVFXODU KHPRJORELQ 0&+f PHDQ FRUSXVFXODU YROXPH 0&9f DQG PHDQ FRUSXVFXODU KHPRJORELQ FRQFHQWUDWLRQ 0&+&f DV D IXQFWLRQ RI VDOLQLW\ IRU &\SULQRGRQ YDULHJDWXV $OO YDOXHV DUH H[SUHVVHG DV PHDQV s VH 6HH WH[W IRU IXUWKHU GHWDLOV RQ EORRG LQGLFHV 6DOLQLW\ SSWf Q +FW bf >+E@ JGOOf 5%& [ PPrf 0&+ SJf 0&9 SPf 0&+& bf s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s s

PAGE 86

)LJXUH 0HDQ HU\WKURF\WH 5%&f FRXQW RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YDULHJDWXV EDUV LQGLFDWH s VH QXPHULFDO YDOXHV DERYH WKH SRLQWV LQ WKH ILJXUH LQGLFDWH VDPSOH VL]H DW HDFK VDOLQLW\f

PAGE 87

6DOLQLW\ SSWf 5HG %ORRG &HOO &RXQW [ O2APP R rf UR &2 R FQ r‘ FQ UR FQ &2 FQ /

PAGE 88

)LJXUH 0HDQ KHPRJORELQ FRQFHQWUDWLRQ >+E@f RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YFLULHJFLWXV EDUV LQGLFDWH s VH QXPHULFDO YDOXHV DERYH WKH SRLQWV LQ WKH ILJXUH LQGLFDWH VDPSOH VL]H DW HDFK VDOLQLW\f

PAGE 89

6DOLQLW\ SSWf +HPRJORELQ &RQFHQWUDWLRQ J GO f UR*RAWQDLVLRRFRR 

PAGE 90

WKURXJK SSW +HPRJORELQ FRQFHQWUDWLRQ ZDV DOVR HOHYDWHG LQ ILVK DFFOLPDWHG WR SSW SSW DQG SSW DOWKRXJK WKHVH YDOXHV ZHUH RQO\ VLJQLILFDQWO\ KLJKHU WKDQ WKH YDOXH RI ILVK DFFOLPDWHG WR SSW +HPDWRFULW PHDVXUHPHQWV VKRZHG OHVV GHSHQGHQFH XSRQ VDOLQLW\ )LJXUH f 0HDQ KHPDWRFULW ZDV KLJKHVW LQ SSW DQG ZDV VLJQLILFDQWO\ GLIIHUHQW WKDQ PHDQ YDOXHV RI ILVK DFFOLPDWHG WR DOO VDOLQLWLHV H[FHSW SSW DQG SSW 1R RWKHU VLJQLILFDQW GLIIHUHQFHV LQ KHPDWRFULW ZHUH QRWHG &DOFXODWHG HU\WKURF\WH LQGLFHV LQGLFDWHG D VOLJKWO\ GLIIHUHQW SDWWHUQ 7KH DYHUDJH FRQFHQWUDWLRQ RI KHPRJORELQ LQ WKH HU\WKURF\WH 0&+&f GLG QRW YDU\ VLJQLILFDQWO\ DPRQJ VDOLQLW\ DFFOLPDWLRQ JURXSV +RZHYHU ERWK WKH DYHUDJH ZHLJKW RI KHPRJORELQ LQ WKH HU\WKURF\WH 0&+f DQG WKH DYHUDJH YROXPH RI WKH HU\WKURF\WH 0&9f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f 7KH FDSDFLWDQFH FRHIILFLHQW UHIOHFWV WKH KHPRJORELQnV R[\JHQ WUDQVSRUWLQJ SURSHUWLHV DQG FDQ EH DGMXVWHG LQ ZKDW KDYH EHHQ WHUPHG nTXDOLWDWLYHn DQG nTXDQWLWDWLYHn ZD\V -HQVHQ f 5HJXODWLRQ RI KHPRJORELQR[\JHQ +E&+f DIILQLW\ UHSUHVHQWV WKH SULPDU\ PHWKRG IRU TXDOLWDWLYHO\ DOWHULQJ R[\JHQ FDUU\LQJ FDSDFLW\ ZLWK FRQWURO RI KHPRJORELQ FRQFHQWUDWLRQ WKH SULPDU\ TXDQWLWDWLYH PHFKDQLVP -HQVHQ f

PAGE 91

)LJXUH 0HDQ KHPDWRFULW +FWf RYHU D UDQJH RI VDOLQLWLHV LQ &\SULQRGRQ YDULHJDWXV EDUV LQGLFDWH s VH QXPHULFDO YDOXHV DERYH WKH SRLQWV LQ WKH ILJXUH LQGLFDWH VDPSOH VL]H DW HDFK VDOLQLW\f

PAGE 92

RR L f L L L r L M L L L L M L L L L L L L L L L L L L L L L L Y U L L L L U L L 6DOLQLW\ SSWf

PAGE 93

%ORRG R[\JHQ FDUU\LQJ FDSDFLW\ FDQ DOVR EH LQFUHDVHG TXDQWLWDWLYHO\ E\ UHOHDVH RI VWRUHG HU\WKURF\WHV E\ DFFHOHUDWLQJ PDWXUDWLRQ RI LPPDWXUH HU\WKURF\WHV DQGRU E\ SURGXFWLRQ RI QHZ HU\WKURF\WHV 0XUDG HW DO f UHOHDVH RI HU\WKURF\WHV IURP VWRUDJH RUJDQV HJ VSOHHQf DSSHDUV WR EH WKH PRVW OLNHO\ VFHQDULR 6RLYLR HW DO :HOOV HW DO f )LVK H[SRVHG WR ZDWHU RI FKDQJLQJ VDOLQLW\ ZRXOG EH H[SHFWHG WR H[SHULHQFH YDULDELOLW\ LQ WKHLU EORRG R[\JHQ FDSDFLWDQFH FRHIILFLHQW DQG EORRG R[\JHQ FDUU\LQJ FDSDFLW\ -HQVHQ HW DO f 4XDQWLWDWLYH PHFKDQLVPV IRU DGMXVWLQJ EORRG R[\JHQ FDUU\LQJ FDSDFLW\ ZHUH H[DPLQHG LQ WKLV VWXG\ )HZ VWXGLHV KDYH H[DPLQHG WKH LQIOXHQFH RI VDOLQLW\ RQ R[\JHQ FDUU\LQJ FDSDFLW\ RI ILVKHV *XHUQVH\ DQG 3ROXKRZLFK f H[DPLQHG WKH EORRG R[\JHQ FDSDFLW\ RI $PHULFDQ HHOV $QJXLOOD URVWUDWDf DFFOLPDWHG WR SSW SSW DQG SSW $V LQ & YDULHJDWXV KHPDWRFULW ZDV KLJKHVW LQ HHOV DFFOLPDWHG WR SSW +RZHYHU ZKLOH R[\JHQ FDSDFLW\ RI DFFOLPDWHG HHOV ZDV KLJKHU LQ SSW WKDQ SSW WKH KLJKHVW R[\JHQ FDSDFLW\ ZDV VHHQ LQ HHOV DFFOLPDWHG WR SSW ,Q D VLPLODU VWXG\ ZLWK WKH FLFKOLG 2UHRFKURPLV QLORWLFXV 6XQ HW DO f REVHUYHG D VLPLODU HIIHFW RI VDOLQLW\ RQ PHDVXUHV RI EORRG R[\JHQ ZLWK KHPRJORELQ FRQFHQWUDWLRQ VLJQLILFDQWO\ KLJKHU LQ SSW WKDQ LQ KLJKHU VDOLQLWLHV WR SSWf 2WKHU IDFWRUV PD\ DOVR FRQWULEXWH WR YDULDWLRQV LQ R[\JHQ FDUU\LQJ FDSDFLW\ RI ILVKHV +DOO DQG *UD\ f ZHUH DPRQJ WKH ILUVW WR QRWH WKDW WKHUH LV D JHQHUDO FRUUHODWLRQ EHWZHHQ WKH KDELWV RI ILVKHV DQG WKH KHPRJORELQ FRQFHQWUDWLRQ RI WKHLU EORRG 0RUH UHFHQW VWXGLHV KDYH VKRZQ WKDW WKLV JHQHUDOL]DWLRQ DOVR DSSOLHV WR HU\ WKURF\WH FRXQW DQG KHPDWRFULW HJ +DZV DQG *RRGQLJKW &REXP /DUVVRQ HW DO 3XWQDP DQG )UHHO f ,Q JHQHUDO KLJKO\ DFWLYH ILVKHV DQG WKRVH WKDW UHJXODUO\ HQFRXQWHU K\SR[LF FRQGLWLRQV KDYH HOHYDWHG EORRG R[\JHQ FDUU\LQJ FDSDFLW\ UHODWLYH WR RWKHU VSHFLHV 9DOXHV GHWHUPLQHG IRU & YDULHJDWXV LQ WKH SUHVHQW VWXG\ DUF FRPSDUDEOH WR RWKHU ILVKHV ZLWK VLPLODU DFWLYLW\ OHYHOV DQG LQGLFDWH WKDW & YDULHJDWXV GRFV QRW SRVVHVV H[FHSWLRQDOO\ KLJK R[\JHQ FDUU\LQJ

PAGE 94

FDSDFLW\ DW DQ\ VDOLQLW\ WHVWHG +DWWLQJK &REXP /DUVVRQ HW DO 3XWQDP DQG )UHHO 6PLW DQG +DWWLQJK 3HOVWHU HW DO Ef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f GXH WR D ODUJHU VXUIDFH DUHD SHU XQLW YROXPH ZKLFK PD\ DOORZ D IDVWHU UDWH RI JDV H[FKDQJH (QHUJHWLF FRQVWUDLQWV UHODWHG WR YLVFRVLW\ RI WKH EORRG PD\ DOVR SOD\ D UROH $ EDODQFH PXVW H[LVW EHWZHHQ WKH DGYDQWDJHV IRU R[\JHQ WUDQVSRUW IURP LQFUHDVHG KHPDWRFULW ZLWK D GLVDGYDQWDJH GXH WR LQFUHDVHG YLVFRVLW\ :HOOV DQG :HEHU f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f 6XUYLYDO LQ IUHVKZDWHU UHTXLUHV PDQ\ RI WKH VDPH UHVSRQVHV WKDW DUF QHFHVVDU\ DW H[WUHPHO\ KLJK VDOLQLWLHV ZLWK ERWK IUHVKZDWHU DQG K\SFUVDOLQF FRQGLWLRQV LPSRVLQJ GLIILFXOW RVPRUHJXODWRU\ SUREOHPV IRU & YDULHJDWXV ,Q ERWK VLWXDWLRQV SUROLIHUDWLRQ RI PLWRFKRQGULD ULFK FHOOV RQ JLOO FSLWKFOLD LV QHHGHG WR PDLQWDLQ LRQLF EDODQFH (YDQV (YDQV :RRG DQG 0DUVKDOO f +RZHYHU

PAGE 95

WKLV H[WHQGV WKH VXUIDFH LQYROYHG LQ LRQLF H[FKDQJH DW WKH H[SHQVH RI JLOO HSLWKHOLD LQYROYHG LQ JDV H[FKDQJH 5HFHQW VWXGLHV KDYH VKRZQ WKDW VXFK SUROLIHUDWLRQ RI PLWRFKRQGULD ULFK FHOOV GRHV LPSDLU UHVSLUDWRU\ JDV WUDQVIHU %LQGRQ HW DK D %LQGRQ HW DO Ef 0HFKDQLVPV WR LQFUHDVH R[\JHQ FDUU\LQJ FDSDFLW\ RI WKH EORRG ZRXOG EH H[SHFWHG XQGHU VXFK FRQGLWLRQV +RZHYHU IUHVKZDWHU FRQGLWLRQV GLIIHU VLJQLILFDQWO\ IURP K\SHUVDOLQH FRQGLWLRQV LQ VHYHUDO ZD\V 0RVW LPSRUWDQWO\ PHWDEROLVP LV UHGXFHG DW H[WUHPH K\SHUVDOLQLWLHV WKLV VWXG\ &KDSWHU f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f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f +RZHYHU DV R[\JHQ QHHGV ZRXOG EH H[SHFWHG WR ULVH DV VDOLQLW\ ZDV LQFUHDVHG RYHU WKLV UDQJH R[\JHQ FDUU\LQJ FDSDFLW\ ZDV H[SHFWHG WR LQFUHDVH DV ZHOO 6HYHUDO SRVVLELOLWLHV PD\ H[SODLQ ZK\ EORRG R[\JHQ FDUU\LQJ FDSDFLW\ GLG QRW LQFUHDVH RYHU WKLV UDQJH RI VDOLQLWLHV )LUVW LQFUHDVHV LQ VDOLQLW\ PD\ UHTXLUH OLWWOH FRPSHQVDWLRQ LQ R[\JHQ FDUU\LQJ FDSDFLW\ XQWLO H[WUHPH K\SFUVDOLQLWLFV DUF UHDFKHG +RZHYHU LW LV PRUH OLNHO\ WKDW WKH ODFN RI D UHVSRQVH PD\ KDYH EHHQ GXH WR WKH IDFW WKDW

PAGE 96

ODUJH YDULDWLRQV LQ VDOLQLW\ DORQH VHOGRP RFFXU XQGHU QDWXUDO FRQGLWLRQV $ ODUJHU UHVSRQVH ZRXOG EH H[SHFWHG LI VDOLQLW\ ZHUH YDULHG WRJHWKHU ZLWK R[\JHQ DQGRU WHPSHUDWXUH $QRWKHU SRVVLELOLW\ LV WKDW & YDULHJDWXV OLNH PDQ\ ILVK VSHFLHV PD\ XWLOL]H PXOWLSOH KHPRJORELQV :HEHU f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f DOWKRXJK WKLV KDV UDUHO\ EHHQ H[DPLQHG ZLWK UHVSHFW WR VDOLQLW\ +RZHYHU VWXGLHV RQ $QJXLOOD DQJXLOOD :HEHU HW DO f DQG 6DOPR VDODU 0£[LPH HW DO f GLG ILQG WKDW LQFUHDVHV LQ VDOLQLW\ EHWZHHQ IUHVKZDWHU DQG VHDZDWHU OHG WR LQFUHDVHV LQ +E&E DIILQLW\ $ KLJK DIILQLW\ KHPRJORELQ PROHFXOH PLJKW DOVR EH DGYDQWDJHRXV XQGHU K\SFUVDOLQH FRQGLWLRQV DOWKRXJK LW PD\ EH LQHIIHFWLYH GXULQJ DFWLYLW\ 0F0DKRQ f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f 6HFRQG WKH SULPDU\ PHFKDQLVP XVHG WR DOWHU +E&E DIILQLW\ UHVXOWV IURP VZHOOLQJ RI HU\WKURF\WHV -HQVHQ :HOOV DQG :HEHU -HQVHQ HW DO f 6ZHOOLQJ RI HU\WKURF\WHV LV QRUPDOO\ DFFRPSDQLHG E\ D ULVH LQ FHOO S+ DQG GHFUHDVHG UHG FHOO KHPRJORELQ 173 DQG $73 FRQFHQWUDWLRQV DOO RI ZKLFK VHUYH WR LQFUHDVH WKH R[\JHQ DIILQLW\ :HOOV DQG :HEHU f +RZHYHU DV VHHQ E\ WKH 0&+& DQG 0&9 YDOXHV GHWHUPLQHG LQ WKLV VWXG\ HU\ WKURF\WHV GLG QRW VZHOO LQ UHVSRQVH WR

PAGE 97

LQFUHDVHG VDOLQLW\ LQ & YDULHJDWXV 7KXV LI +E DIILQLW\ LV DOWHUHG ZLWK FKDQJHV LQ HQYLURQPHQWDO VDOLQLW\ LW PXVW EH FKDQJHG LQ VRPH RWKHU PDQQHU 7KLV VWXG\ FOHDUO\ LQGLFDWHV WKDW VDOLQLW\ GRHV LQIOXHQFH WKH R[\JHQ FDUU\LQJ FDSDFLW\ RI WKH EORRG RI & YDULHJDWXV 4XDQWLWDWLYH GLIIHUHQFHV LQ KHPRJORELQ FRQFHQWUDWLRQ KHPDWRFULW DQG HU\WKURF\WH FRXQW ZHUH QRWHG LQ UHVSRQVH WR FKDQJLQJ VDOLQLW\ $V GLVFXVVHG DERYH LW DOVR VHHPV OLNHO\ WKDW VDOLQLW\f PD\ LQIOXHQFH TXDOLWDWLYH FKDQJHV LQ EORRG R[\JHQ WUDQVSRUW LQ & YDULHJDWXV )XUWKHU UHVHDUFK LV QHHGHG WR EHWWHU XQGHUVWDQG WKH LQIOXHQFH RI VDOLQLW\ RQ EORRG R[\JHQ OHYHOV LQ HXU\KDOLQH WHOHRVWV

PAGE 98

&+$37(5 6800$5< $1' &21&/86,216 7KLV VWXG\ H[DPLQHG FRVWV DVVRFLDWHG ZLWK OLIH RI D WHOHRVW LQ D YDULDEOH VDOLQLW\ HQYLURQPHQW UHSUHVHQWHG KHUH E\ D VDOW PDUVK &\SULQRGRQ YDULHJDWXV ZDV XVHG WR H[DPLQH WKH LQIOXHQFH RI VDOLQLW\ RQ URXWLQH PHWDEROLF UDWH 505f FULWLFDO R[\JHQ WHQVLRQ 3Ff RVPRUHJXODWLRQ DQG EORRG R[\JHQ FDUU\LQJ FDSDFLW\ 5HVXOWV DUH VXPPDUL]HG EHORZ f )LHOG PHDVXUHPHQWV LQ WKH &HGDU .H\ VDOW PDUVK LQGLFDWHG WKDW WKLV KDELWDW XQGHUJRHV H[WHQVLYH YDULDWLRQ LQ VDOLQLW\ WHPSHUDWXUH DQG R[\JHQ f 505 ZDV UHODWLYHO\ FRQVWDQW RYHU D UDQJH RI VDOLQLWLHV IURP SSW WR SSW $W KLJKHU VDOLQLWLHV 505 EHJDQ WR GHFOLQH DQG ZDV VLJQLILFDQWO\ GHSUHVVHG XQGHU K\SFUVDOLQH FRQGLWLRQV f )ROORZLQJ VHTXHQWLDO DFFOLPDWLRQ WR H[SHULPHQWDO VDOLQLWLHV 3F ZDV XQDIIHFWHG E\ FKDQJHV LQ VDOLQLW\ EHWZHHQ SSW DQG SSW ZLWK 3F LQFUHDVLQJ DW KLJKHU VDOLQLWLHV f 5HGXFWLRQ LQ PHWDEROLVP DQG ULVH LQ 3F FRUUHVSRQGHG ZHOO ZLWK D UHGXFHG DELOLW\ RI & YDULHJDWXV WR UHJXODWH SODVPD RVPRODOLW\ HIILFLHQWO\ 2VPRWLF SHUPHDELOLW\ RI WKH JLOOV PD\ EH UHGXFHG DW KLJK VDOLQLWLHV WR RIIVHW RVPRWLF ORVVHV RU LRQLF JDLQV WRIURP WKH HQYLURQPHQW LQGLUHFWO\ UHGXFLQJ WKH SRWHQWLDO IRU R[\JHQ XSWDNH DV ZHOO f 9DULDWLRQV LQ 505 DQG 3F DV D IXQFWLRQ RI HQYLURQPHQWDO VDOLQLW\ REVHUYHG LQ WKLV VWXG\ VXJJHVW WKDW & YDULHJDWXV UHVSRQGV WR KLJK VDOLQLWLHV E\ UHGXFLQJ HQHUJ\ H[SHQGLWXUHV 7KHVH UHVSRQVHV HIIHFWLYHO\ LQFUHDVHV WKH WLPH & YDULHJDWXV FDQ WROHUDWH VXFK FRQGLWLRQV DOEHLW DW D FRVW RI D UHGXFWLRQ LQ HQHUJHWLF SURFHVVHV 7KLV VWUDWHJ\ ILWV WKH FRQFHSW RI VFRSH IRU VXUYLYDO DV GHVFULEHG E\ +RFKDFKND f f :KHQ & YDULHJDWXV ZDV H[SRVHG WR VLPXODWHG WLGDO FKDQJHV LQ VDOLQLW\ 505 ZDV XQDIIHFWHG LQ VDOLQLW\ WULDOV ZKHUH ERWK DFFOLPDWLRQ DQG ILQDO VDOLQLWLHV ZHUH LQ WKH UDQJH

PAGE 99

W\SLFDOO\ HQFRXQWHUHG E\ WKLV SRSXODWLRQ LQ LWV QDWLYH KDELWDW :KHUH WKH DFFOLPDWLRQ RU ILQDO VDOLQLWLHV ZHUH H[WUHPHO\ KLJK DQG SSWf RU H[WUHPHO\ ORZ SSWf 505 ZDV GHSUHVVHG f $FFOLPDWLRQ VWDWH ZDV WKH PRVW LPSRUWDQW IDFWRU GHWHUPLQLQJ WKH PHWDEROLF UHVSRQVH WR VLPXODWHG WLGDO FKDQJHV LQ VDOLQLW\ +RZHYHU GLUHFWLRQ RI WKH VDOLQLW\ FKDQJH DOVR LQIOXHQFHG PHWDEROLVP LQ & YDULHJDWXV ZLWK LQFUHDVLQJ VDOLQLW\ GHDOW ZLWK PRUH HIILFLHQWO\ WKDQ GHFUHDVLQJ VDOLQLW\ f 6LPXODWHG WLGDO H[SHULPHQWV FRUURERUDWH WKH K\SRWKHVLV WKDW & YDULHJDWXV WROHUDWHV H[WUHPHV LQ VDOLQLW\ E\ ORZHULQJ PHWDEROLVP DQG KHQFH GHFUHDVLQJ HQHUJ\ H[SHQGLWXUHV )ROORZLQJ DGYHUVH FRQGLWLRQV PHWDEROLVP UHWXUQV WR QRUPDO OHYHOV f &\SULQRGRQ YDULHJDWXV LV DQ H[FHOOHQW UHJXODWRU RI SODVPD RVPRODOLW\ HYHQ ZKHQ H[SRVHG WR ODUJH IOXFWXDWLRQV LQ VDOLQLW\ ZLWKLQ WKH UDQJH RI VDOLQLWLHV W\SLFDOO\ HQFRXQWHUHG 'DLO\ IOXFWXDWLRQV LQ VDOLQLW\ RI XS WR SSW HOLFLWHG QR VLJQLILFDQW GLIIHUHQFHV LQ RVPRUHJXODWRU\ DELOLW\ ZKHQ FRPSDUHG WR FRQWURO ILVK f 3ULRU H[SRVXUH WR IOXFWXDWLRQV LQ VDOLQLW\ GRHV LPSDUW DQ RVPRUHJXODWRU\ DGYDQWDJH )LVKHV SUHYLRXVO\ H[SRVHG WR ODUJH IOXFWXDWLRQV LQ VDOLQLW\ UHJXODWHG SODVPD RVPRODOLW\ EHWWHU WKDQ ILVKHV WKDW KDG SUHYLRXVO\ H[SHULHQFHG QR RU VPDOO FKDQJHV LQ VDOLQLW\ ,QFUHDVLQJ VDOLQLW\ KDG D JUHDWHU LPSDFW RQ UHJXODWLRQ RI SODVPD RVPRODOLW\ WKDQ GLG GHFUHDVHV LQ VDOLQLW\ f 6DOLQLW\ KDG D VLJQLILFDQW HIIHFW RQ EORRG R[\JHQ FDUU\LQJ FDSDFLW\ LQ & YDULHJDWXV DOWKRXJK GLIIHUHQFHV ZHUH RQO\ QRWHG DW WKH YHU\ KLJKHVW WR SSWf DQG ORZHVW SSWf VDOLQLWLHV WHVWHG 2[\JHQ FDUU\ LQJ FDSDFLW\ DQG DOO EORRG LQGLFHV ZHUH KLJKHVW LQ WKH JURXS DFFOLPDWHG WR SSW f & YDULHJDWXV H[KLELWHG OLWWOH FKDQJH LQ R[\JHQ FDUU\LQJ FDSDFLW\ RYHU WKH UDQJH RI VDOLQLWLHV EHWZHHQ SSW DQG SSW 3RVVLEOH UHDVRQV IRU WKLV LQFOXGH Df LQFUHDVHV LQ VDOLQLW\ PD\ UHTXLUH OLWWOH FRPSHQVDWLRQ LQ R[\JHQ FDUU\LQJ FDSDFLW\ Ef GHWHFWDEOH FKDQJHV PD\ RQO\ RFFXU ZKHQ VDOLQLW\ LV YDULHG LQ FRQMXQFWLRQ ZLWK YDULDWLRQV LQ R[\JHQ DQGRU

PAGE 100

WHPSHUDWXUH Ff & YDULHJDWXV PD\ XWLOL]H PXOWLSOH KHPRJORELQV DQGRU Gf WKH SULPDU\ PHFKDQLVP WR LQFUHDVH R[\JHQ FDUU\LQJ FDSDFLW\ PD\ LQVWHDG EH WKURXJK DGMXVWPHQW RI +E DIILQLW\ f (U\WKURF\WH FRXQW ZDV WKH PRVW FRQVLVWHQW DQG KHPDWRFULW WKH OHDVW FRQVLVWHQW PHDVXUH RI WKH LQIOXHQFH RI VDOLQLW\ RQ EORRG R[\JHQ OHYHO &RPSHWLWLRQ RU SUHGDWLRQ SUHVVXUH PD\ EH OHVV LQWHQVH LQ KDUVK IOXFWXDWLQJ HQYLURQPHQWV DQG WKDW FHUWDLQ VSHFLHV PD\ DYRLG WKHVH SUHVVXUHV E\ HYROXWLRQ RI ZLGH SK\VLFRFKHPLFDO WROHUDQFHV DQG WKH XVH RI VXFK HQYLURQPHQWV 0DWWKHZV DQG 6W\URQ f &\SULQRGRQ YDULHJDWXV VHHPV WR ILW WKLV PROG ZHOO DV WKLV VSHFLHV DSSHDUV WR EH D JHQHUDOLVW WKDW YHU\ VXFFHVVIXOO\ LQKDELWV KDUVK DQG YDULDEOH KDELWDWV ZKHUH LW GRHV QRW KDYH WR EH YHU\ HIILFLHQW WR FRPSHWH ZLWK RWKHU VSHFLHV RI ILVKHV 0DUWLQ %HUU\ f 7KLV DUJXPHQW PD\ H[SODLQ ZK\ & YDULHJDWXV GRHV QRW LQYDGH IUHVKZDWHU LQ PRUH ORFDOHV DQG ZK\ WKH\ DUH QRW YHU\ DEXQGDQW LQ PRVW IUHVKZDWHU V\VWHPV H[FHSW LQ VRXWK )ORULGD IUHVKZDWHUV ZKHUH WHPSHUDWXUH DQG GLVVROYHG R[\JHQ DUF YDULDEOH DQG H[WUHPH )XUWKHUPRUH LW LV EHOLHYHG WKDW & YDULHJDWXV KDV FRQTXHUHG LWV ZLGH JHRJUDSKLF UDQJH E\ SK\VLRORJLFDO IOH[LELOLW\ UDWKHU WKDQ E\ ORFDO DFFRPPRGDWLRQ LQ SK\VLRORJ\ RU OLIH KLVWRU\ %HUU\ f 7KLV K\SRWKHVLV UHPDLQ WR EH ULJRURXVO\ WHVWHG DOWKRXJK WKLV VWXG\ OHQGV IXUWKHU HYLGHQFH IRU WKH SK\VLRORJLFDO IOH[LELOLW\ RI & YDULHJDWXV ([DPLQDWLRQ RI WKH SK\VLRORJLFDO DELOLW\ RI RWKHU SRSXODWLRQV RI & YDULHJDWXV PD\ KHOS WR UHVROYH WKLV LVVXH 7KH VXFFHVV RI D ILVK VSHFLHV LQ D VWUHVVIXO HQYLURQPHQW PD\ GHSHQG SULPDULO\ XSRQ WKH SURSRUWLRQ RI WKH SRSXODWLRQ WKDW VXUYLYHV WKH DGYHUVH FRQGLWLRQV ZLWK HYHQ VKRUWn WHUP LQFUHDVHV LQ WROHUDQFH VLJQLILFDQW 0DWWKHZV DQG 6W\URQ f 7KLV K\SRWKHVLV DSSHDUV WR EH UHOHYDQW WR WKH UHVXOWV RI WKH SUHVHQW VWXG\ &\SULQRGRQ YDULHJDWXV LV ZHOO DGDSWHG WR D YDU\LQJ VDOLQLW\ HQYLURQPHQW ,WV PHWDEROLF UDWH 3F DQG DELOLW\ WR HIILFLHQWO\ RVPRUHJXODWF LV XQDIIHFWHG E\ FKDQJHV LQ VDOLQLW\ RYHU WKH W\SLFDO UDQJH HQFRXQWHUHG HYHQ ZKHQ VDOLQLW\ LV FKDQJHG YHU\ UDSLGO\ )XUWKHUPRUH & YDULHJDWXV DSSHDUV WR WROHUDWH H[WUHPHV LQ VDOLQLW\ E\ GHFUHDVLQJ HQHUJ\ H[SHQGLWXUHV ZDLWLQJ IRU FRQGLWLRQV WR LPSURYH

PAGE 101

DQG WKHQ UHVSRQGLQJ E\ LQFUHDVLQJ PHWDEROLVP EDFN WR QRUPDO OHYHOV 7KLV LV D SRWHQWLDOO\ DGDSWLYH UHVSRQVH IRU OLIH LQ D YDULDEOH VDOLQLW\ HQYLURQPHQW :KLOH WKLV VWXG\ DQVZHUV PDQ\ TXHVWLRQV UHODWLQJ WR WKH SK\VLRORJ\ RI VDOLQLW\ DGDSWDWLRQ LQ & YDULHJDWXV LW DOVR UDLVHV QHZ RQHV &KLHI DPRQJ WKHVH LV WKH TXHVWLRQ RI KRZ UHVSRQVHV GXH WR VDOLQLW\ ZRXOG EH LQIOXHQFHG E\ LQWHUDFWLRQV ZLWK FKDQJHV LQ WHPSHUDWXUH DQGRU GLVVROYHG R[\JHQ )RU H[DPSOH & YDULHJDWXV QRUPDOO\ H[SHULHQFHV KLJK VDOLQLWLHV WRJHWKHU ZLWK KLJK WHPSHUDWXUHV 7KH LQWHUDFWLRQ RI WKHVH WZR SDUDPHWHUV RQ RVPRUHJXODWLRQ PHWDEROLVP DQG R[\JHQ FDUU\LQJ FDSDFLW\ ZRXOG EH D QDWXUDO FRQWLQXDWLRQ RI WKH FXUUHQW VWXG\ $QRWKHU DUHD RI IXWXUH UHVHDUFK ZRXOG LQFOXGH DQ H[DPLQDWLRQ LQWR WKH SRVVLEOH DELOLW\ RI & YDULHJDWXV WR XWLOL]H DQDHURELF PHWDEROLVP XQGHU YDU\LQJ VDOLQLW\ UHJLPHV DV LW ZDV VXJJHVWHG E\ 6XEUDKPDQ\DP f WKDW VDOW PDUVK ILVKHV PD\ XWLOL]H DQDHURELF PHWDEROLVP PRUH WKDQ RWKHU JURXSV RI ILVKHV ,W KDV DOVR EHHQ SURSRVHG WKDW SHUPHDELOLW\ FKDQJHV RI WKH JLOO PD\ OHDG WR UHGXFWLRQV LQ PHWDEROLVP DW KLJK VDOLQLWLHV 1RUGOLH HW DO f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f WKDW ILVK WKDW DUH FDSDEOH RI ZLWKVWDQGLQJ VXGGHQ DQG IUHTXHQW VDOLQLW\ FKDQJHV DUH JHQHUDOO\ VSHFLDOL]HG PRUSKRORJLFDOO\ DQG SK\VLRORJLFDOO\ LQ ZD\V WKDW UHVWULFW WKHLU OLIHVW\OH VHHPV HUURQHRXV ZKHQ ILVKHV VXFK DV & YDULHJDWXV DUH FRQVLGHUHG :KLOH IHZ GDWD H[LVW RQ WKH UHVSRQVHV RI RWKHU ILVKHV WR ZLGH UDQJHV LQ VDOLQLW\ WKH SDWWHUQV VHHQ LQ & YDULHJDWXV PD\ UHSUHVHQW D JHQHUDO SDWWHUQ IRU ILVKHV LQKDELWLQJ YDULDEOH VDOLQLW\ HQYLURQPHQWV

PAGE 102

$33(1',; &5,7,&$/ 2;<*(1 7(16,21 ),*85(6 &ULWLFDO R[\JHQ WHQVLRQ 3Ff ILJXUHV ZHUH SORWWHG IRU HDFK ILVK XVHG LQ WKH VWXG\ GHVFULEHG LQ FKDSWHU 7KHVH SORWV FRQVLVWHG RI ZHLJKWDGMXVWHG R[\JHQ FRQVXPSWLRQ UDWHV PJ 2 KnOf SORWWHG DJDLQVW R[\JHQ WHQVLRQ PP +Jf GXULQJ WKH LQWHUYDO RYHU ZKLFK R[\JHQ FRQVXPSWLRQ ZDV FDOFXODWHG %HFDXVH ILVK ZHUH XVHG LQ WKLV VWXG\ LW LV LPSUDFWLFDO WR VKRZ HDFK ILJXUH )LJXUHV $ $ DQG $ DUH UHSUHVHQWDWLYH VDPSOHV RI WKHVH ILJXUHV *HQHUDOL]HG 3F ILJXUHV ZHUH SURGXFHG IRU HDFK VDOLQLW\ JURXS IURP WKH PHDQ 3F PHDQ URXWLQH PHWDEROLF UDWH 505f DQG PHDQ VORSH LQ WKH FRQIRUPDWLRQ UHJLRQ 7KHVH DUH VKRZQ LQ ILJXUH $

PAGE 103

)LJXUH $O 3ORW LQGLFDWLQJ WKH FDOFXODWLRQ RI WKH FULWLFDO R[\JHQ WHQVLRQ 3Ff IRU DQ LQGLYLGXDO &\SULQRGRQ YDULHJDWXV LQ ZDWHU DW SSW

PAGE 104

2[\JHQ 7HQVLRQ PP +Jf $GMXVWHG 5RXWLQH 0HWDEROLF 5DWH PJ 2[\JHQKUf R R R R R r R .! Mr LQ ER R H

PAGE 105

)LJXUH $ 3ORW LQGLFDWLQJ WKH FDOFXODWLRQ RI WKH FULWLFDO R[\JHQ WHQVLRQ 3Ff IRU DQ LQGLYLGXDO &\SULQRGRQ YFLULHJDWXV LQ ZDWHU DW SSW

PAGE 106

$GMXVWHG 5RXWLQH 0HWDEROLF 5DWH PJ 2[\JHQKUf ‘ W f§ ‘ If ‘‘ n U L } L , U ‘> ‘ } } U L 2[\JHQ 7HQVLRQ PP +Jf

PAGE 107

)LJXUH $ 3ORW LQGLFDWLQJ WKH FDOFXODWLRQ RI WKH FULWLFDO R[\JHQ WHQVLRQ 3Ff IRU DQ LQGLYLGXDO &\SULQRFORQ YFLULHJDWXV LQ ZDWHU DW ;f SSW

PAGE 108

$GMXVWHG 5RXWLQH 0HWDEROLF 5DWH PJ 2[\JHQKUf 3F PP +J 7 n n 7 7 , ) 7f§f§ , ‘ W 2[\JHQ 7HQVLRQ PP +Jf

PAGE 109

)LJXUH $ *HQHUDOL]HG FULWLFDO R[\JHQ WHQVLRQ 3Ff SORWV DW HDFK VDOLQLW\ XVHG LQ WKH 3F H[SHULPHQWV 3ORWV ZHUH SURGXFHG E\ XVLQJ WKH PHDQ 3F PHDQ URXWLQH PHWDEROLF UDWH 505f DQG PHDQ VORSH LQ WKH FRQIRUPDWLRQ UHJLRQ IRU HDFK VDOLQLW\ JURXS

PAGE 110

2[\JHQ 7HQVLRQ PP +Jf 2[\JHQ 7HQVLRQ PP +Jf 2[\JHQ &RQVXPSWLRQ 5DWH PJ 2[\JHQKUf RSSRRSSRAA} 1ZL£LEVERLREA1 2[\JHQ &RQVXPSWLRQ 5DWH PJ 2[\JHQKUf RRRSRRSR f§ 1ZALLQAF'EA1 2[\JHQ &RQVXPSWLRQ 5DWH PJ 2[\JHQKUf SSSSSSSSArArr .fZAZMf0FREEnrNf 2[\JHQ &RQVXPSWLRQ 5DWH PJ 2[\JHQKUf RRSRRSSRrAA .MZ9LPnVLFRLREA1

PAGE 111

2[\JHQ 7HQVLRQ PP +Jf 2[\JHQ 7HQVLRQ PP +Jf SRQXQXR' S?\ RMQ6\ 2[\JHQ &RQVXPSWLRQ 5DWH PJ 2[\JHQKUf r rr 1ZLOQ•OV•RLREA1 2[\JHQ &RQVXPSWLRQ 5DWH PJ 2[\JHQKUf }} r 1:$ZEO6•RLRE1 2[\JHQ &RQVXPSWLRQ 5DWH PJ 2[\JHQKUf 2 2 2 2 2 2 2 .f FR ,Q A ER 3 r r r LR E A 1 2[\JHQ &RQVXPSWLRQ 5DWH PJ 2[\JHQKUf }}} NLZLXLEL1LFRLRRA1

PAGE 112

SRQXQXRR f§ S?9 -Q6\ 2[\JHQ &RQVXPSWLRQ 5DWH PJ 2[\JHQKUf 22222222rrr .f/MWN/QF'1LERe!fr?f 2[\JHQ &RQVXPSWLRQ 5DWH PJ 2[\JHQKUf SRSRSRSSrrr .f 8f L FQ f fVL ER OR E UR 2[\JHQ &RQVXPSWLRQ 5DWH PJ 2[\JHQKUf SSSSSSSSArrr 1:A/QLMO1O&'FEn1 72,

PAGE 113

$33(1',; +(/' 0($685(0(176 )LHOG PHDVXUHPHQWV ZHUH WDNHQ LQ WKH &HGDU .H\ DUHD IURP -XQH WKURXJK -XQH 7KHVH PHDVXUHPHQWV ZHUH WDNHQ DW WZR GHSWKV ERWWRP DQG VXUIDFHf IRU HDFK RI IRXU VLWHV LQ WKH DUHD ZKHQHYHU SRVVLEOH 7KH YDULDEOHV PHDVXUHG ZHUH R[\JHQ FRQFHQWUDWLRQ PJ /nOf VDOLQLW\ SSWf DQG WHPSHUDWXUH r&f )LJXUH $ VKRZV WKH UHODWLRQVKLSV EHWZHHQ WKHVH WKUHH YDULDEOHV DW HDFK VLWH DQG GHSWK RYHU WKH FRXUVH RI WKH \HDU 2[\JHQ FRQFHQWUDWLRQ ZDV KLJKO\ LQYHUVHO\ FRUUHODWHG ZLWK FKDQJHV LQ ERWK VDOLQLW\ DQG WHPSHUDWXUH 6DOLQLW\ VHHPV WR EH PRVW KLJKO\ FRUUHODWHG ZLWK FKDQJLQJ R[\JHQ OHYHOV DOWKRXJK VDOLQLW\ DQG WHPSHUDWXUH DSSHDU WR EH OLQNHG DV ZHOO DOEHLW ZLWK PRUH VLWH WR VLWH YDULDWLRQ

PAGE 114

)LJXUH $ 2[\JHQ FRQFHQWUDWLRQ PJ /nOf VDOLQLW\ SSWf DQG WHPSHUDWXUH r&f DW IRXU VLWHV LQ WKH &HGDU .H\ DUHD WDNHQ EHWZHHQ -XQH DQG -XQH 9DOXHV JLYHQ DV PHDQV EDUV LQGLFDWH s VH Df 0HDVXUHPHQWV WDNHQ RQ WKH ERWWRP DW VLWH Ef PHDVXUHPHQWV WDNHQ RQ WKH VXUIDFH DW VLWH Ff 0HDVXUHPHQWV WDNHQ RQ WKH ERWWRP DW VLWH Gf PHDVXUHPHQWV WDNHQ RQ WKH VXUIDFH DW VLWH Hf 0HDVXUHPHQWV WDNHQ RQ WKH ERWWRP DW VLWH PHDVXUHPHQWV WDNHQ RQ WKH VXUIDFH DW VLWH Jf 0HDVXUHPHQWV WDNHQ RQ WKH VXUIDFH DW VLWH

PAGE 115

3DUW D f§2[\JHQ PJf 7HPSHUDWXUH &f $ 6DOLQLW\ SSWf 3DUW E :LQWHU

PAGE 116

3DUW F 2[\JHQ PJf 7HPSHUDWXUH &f 6DOLQLW\ SSWf 3DUW G )LJXUH $ f§ FRQWLQXHG

PAGE 117

3DUW H 2[\JHQ PJf ff 7HPSHUDWXUH &f 6DOLQLW\ SSWf 3DUW I )LJXUH $ FRQWLQXHG

PAGE 118

3DUW J 2[\JHQ PJf 7HPSHUDWXUH &f 6DOLQLW\ SSWf )LJXUH $ FRQWLQXHG

PAGE 119

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f ELRUHJXODWRU RI PRVTXLWR ODUYDH RI $HGHV WDHQLRUK\QFKXV DQG &XOH[ EDKDPHQVLV LQ WKH ,VOH RI
PAGE 120

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f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f PHWDEROLF DQG UHVSLUDWRU\ DGDSWDWLRQV 5HVSLUDWLRQ 3K\VLRORJ\ %UHWW -5 DQG 7'' *URYHV 3K\VLRORJLFDO HQHUJHWLFV 3DJHV LQ :6 +RDU '5DQGDOO DQG -5 %UHWW HGLWRUV )LVK SK\VLRORJ\ 9ROXPH $FDGHPLF 3UHVV 1HZ
PAGE 121

&DPSEHOO 7 DQG ) 0XUUX $Q LQWURGXFWLRQ WR ILVK KHPDWRORJ\ &RPSHQGLXP RI &RQWLQXLQJ (GXFDWLRQ IRU WKH 3UDFWLFLQJ 9HWHULQDULDQ &HFK --U 5HVSLURPHWU\n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
PAGE 122

,OO 'RQQHOO\ DQG -7RUUHV 2[\JHQ FRQVXPSWLRQ RI PLGZDWHU ILVKHV DQG FUXVWDFHDQV IURP WKH HDVWHUQ *XOI RI 0H[LFR 0DULQH %LRORJ\ 'XJJLQV &) -U $$ .DUOLQ DQG .* 5HO\HD (OHFWURSKRUHWLF FRPSDULVRQ RI &\SULQRGRQ YDULHJDWXV /DFSHGH DQG &\SULQRGRQ KXEEVL &DUU ZLWK FRPPHQWV RQ WKH JHQXV &\SULQRGRQ $WKHULQLIRUPHV &\SULQRGRQWLGDHf 1RUWKHDVW *XOI 6FLHQFH 'XQVRQ :$ 3 )ULFDQR DQG :6DGLQVNL 9DULDWLRQ LQ WROHUDQFH WR DELRWLF VWUHVVHV DPRQJ V\PSDWULF VDOW PDUVK ILVK :HWODQGV 'XQVRQ :$ DQG 7UDYLV 3DWWHUQV LQ WKH HYROXWLRQ RI SK\VLRORJLFDO VSHFLDOL]DWLRQ LQ VDOWPDUVK DQLPDOV (VWXDULHV 'Z\HU 0 DQG $ %HXOLJ 6RFLDO H[SHULHQFH DQG WKH GHYHORSPHQW RI DJJUHVVLYH EHKDYLRU LQ WKH SXSILVK &\SULQRGRQ YDULHJDWXVf -RXUQDO RI &RPSDUDWLYH 3V\FKRORJ\ (FKHOOH $$ DQG 3&RQQRU 5DSLG JHRJUDSKLFDOO\ H[WHQVLYH JHQHWLF LQWURJUHVVLRQ DIWHU VHFRQGDU\ FRQWDFW EHWZHHQ WZR SXSILVK VSHFLHV &\SULQRGRQ &\SULQRGRQWLGDHf (YROXWLRQ (FKHOOH $$ DQG $) (FKHOOH (YROXWLRQDU\ UHODWLRQVKLSV DPRQJ LQODQG SXSILVKFV RI WKH &\SULQRGRQ YDULHJDWXV FRPSOH[ 3DJH LQ (3 3LVWHU HGLWRU 3URFHHGLQJV RI WKH 'HVHUW )LVKHV &RXQFLO 9ROXPH 'HVHUW )LVKHV &RXQFLO /DV 9HJDV 19 (FKHOOH $) DQG $$ (FKHOOH $VVHVVPHQW RI JHQHWLF LQWURJUHVVLRQ EHWZHHQ WZR SXSILVK VSHFLHV &\SULQRGRQ HOHJDQV DQG & YDULHJDWXV &\SULQRGRQWLGDHf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

PAGE 123

)HUUDULV 53 -0 $OPHQGUDV DQG $3 -D]XO &KDQJHV LQ SODVPD RVPRODOLW\ DQG FKORULGH FRQFHQWUDWLRQ GXULQJ DEUXSW WUDQVIHU RI PLONILVK &KDQRV FKDQRVf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
PAGE 124

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nV VFRSH IRU DFWLYLW\ 7UDQVDFWLRQV RI WKH $PHULFDQ )LVKHULHV 6RFLHW\ +ROPHV :1 DQG (0 'RQDOGVRQ 7KH ERG\ FRPSDUWPHQWV DQG WKH GLVWULEXWLRQ RI HOHFWURO\WHV 3DJHV LQ :6 +RDU DQG '5DQGDOO HGLWRUV )LVK SK\VLRORJ\ 9ROXPH $FDGHPLF 3UHVV 1HZ
PAGE 125

-RKDQVHQ DQG 5( :HEHU 2Q WKH DGDSWDELOLW\ RI KHPRJORELQ IXQFWLRQ WR HQYLURQPHQWDO FRQGLWLRQV 3DJHV LQ 3 6SHQFHU 'DYLHV HGLWRU 3HUVSHFWLYHV LQ H[SHULPHQWDO ELRORJ\ 9ROXPH =RRORJ\ 3HUJDPRQ 3UHVV 2[IRUG -RKQVRQ :( 0RUSKRORJLFDO YDULDWLRQ DQG ORFDO GLVWULEXWLRQ RI &\SULQRGRQ YDULHJDWXV LQ )ORULGD 3K' 'LVVHUWDWLRQ )ORULGD 7HFKQRORJLFDO 8QLYHUVLW\ 2UODQGR )/ S -RKQVWRQ &( DQG -& &KHYHULH &RPSDUDWLYH DQDO\VLV RI LRQRUHJXODWLRQ LQ UDLQERZ WURXW 6DOPR JDLUGQHULf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f DGDSWDWLRQV WR FKDQJLQJ HQYLURQPHQWV 3DJHV LQ 51DLPDQ DQG '/ 6ROW] HGLWRUV )LVKHV LQ 1RUWK $PHULFDQ GHVHUWV -RKQ :LOH\ DQG 6RQV 1HZ
PAGE 126

.OW] DQG + 2QNHQ /RQJWHUP DFFOLPDWLRQ RI WKH WHOHRVW 2UHRFKURPLV PRVVDPELFXV WR YDULRXV VDOLQLWLHV WZR GLIIHUHQW VWUDWHJLHV LQ PDVWHULQJ K\SHUWRQLF VWUHVV 0DULQH %LRORJ\ .XVKODQ -$ 3RSXODWLRQ IOXFWXDWLRQV RI HYHUJODGHV ILVKHV &RSHLD /DUVVRQ $ 0O -RKDQVVRQ6MREHFN DQG 5 )£QJH &RPSDUDWLYH VWXG\ RI VRPH KDHPDWRORJLFDO DQG ELRFKHPLFDO EORRG SDUDPHWHUV LQ ILVKHV IURP WKH 6NDJHUUDN -RXUQDO RI )LVK %LRORJ\ /HORXS DQG -0 /HEHO 7ULLRGRWK\URQLQH LV QHFHVVDU\ IRU WKH DFWLRQ RI JURZWK KRUPRQH LQ DFFOLPDWLRQ WR VHDZDWHU RI EURZQ WURXW 6DOPR WUXWWDf DQG UDLQERZ WURXW 2QFRUK\QFKXV UQ\NLVVf )LVK 3K\VLRORJ\ DQG %LRFKHPLVWU\ /LQWRQ 7. 6DOLQLW\ DQG WHPSHUDWXUH HIIHFWV RQ WKH FKURQLF WR[LFLW\ RI GLQLWURSKHQRO DQG QLWURSKHQRO WR VKHHSVKHDG PLQQRZV &\SULQRGRQ YDULHJDWXVf 0DVWHUV 7KHVLV 7KH 8QLYHUVLW\ RI :HVW )ORULGD 3HQVDFROD )/ S /RWDQ 5 DQG ( 6NDGKDXJH ,QWHVWLQDO VDOW DQG ZDWHU WUDQVSRUW LQ D HXU\KDOLQH WHOHRVW $SKDQLXV GLVSDU &\SULQRGRQWLGDHf &RPSDUDWLYH %LRFKHPLVWU\ DQG 3K\VLRORJ\ $ /RZH &+ '6 +LQGV DQG ($ +DOSHP ([SHULPHQWDO FDWDVWURSKLF VHOHFWLRQ DQG WROHUDQFHV WR ORZ R[\JHQ FRQFHQWUDWLRQ LQ QDWLYH $UL]RQD IUHVKZDWHU ILVKHV (FRORJ\ 0DGDQ 0RKDQ 5DR 2[\JHQ FRQVXPSWLRQ RI UDLQERZ WURXW 6DOPR JDLUGQHULf LQ UHODWLRQ WR DFWLYLW\ DQG VDOLQLW\ &DQDGLDQ -RXUQDO RI =RRORJ\ 0DGDQ 0RKRQ 5DR ,QIOXHQFH RI DFWLYLW\ DQG VDOLQLW\ RQ WKH ZHLJKWGHSHQGHQW R[\JHQ FRQVXPSWLRQ RI WKH UDLQERZ WURXW 6DOPR JDLUGQHUL 0DULQH %LRORJ\ 0DQFHUD -0 -0 3HUH])LJDUHV DQG 3 )HPDQGF]/LHEUH] 2VPRUHJXODWRU\ UHVSRQVHV WR DEUXSW VDOLQLW\ FKDQJHV LQ WKH HXU\KDOLQH JLOWKHDG VHD EUHDP 6SDUXV DXUDWD /f &RPSDUDWLYH %LRFKHPLVWU\ DQG 3K\VLRORJ\ $ 0DUWLQ )' ,QWUDVSHFLILF YDULDWLRQ LQ RVPRWLF DELOLWLHV RI &\SULQRGRQ YDULHJDWXV /DFFSFGH IURP WKH 7H[DV &RDVW (FRORJ\ 0DUWLQ )' )HHGLQJ KDELWV RI &\SULQRGRQ YDULHJDWXV &\SULQRGRQWLGDHf IURP WKH 7H[DV FRDVW 6RXWKZHVWHUQ 1DWXUDOLVW 0DUWLQ )' )DFWRUV LQIOXHQFLQJ ORFDO GLVWULEXWLRQ RI &\SULQRGRQ YDULHJDWXV 3LVFHV &\SULQRGRQWLGDHf 7UDQVDFWLRQV RI WKH $PHULFDQ )LVKHULHV 6RFLHW\ 0DWWKHZV :DQG -7 6W\URQ -U 7ROHUDQFH RI KHDGZDWHU YV PDLQVWUHDP ILVKHV IRU DEUXSW SK\VLFRFKHPLFDO FKDQJHV $PHULFDQ 0LGODQG 1DWXUDOLVW 0£[LPH 9 0 3F\UDXG:DLW]HQFJJFU &ODLUFDX[ DQG & 3F\UDXG (IIHFWV RI UDSLG WUDQVIHU IURP VHD ZDWHU WR IUHVK ZDWHU RQ UHVSLUDWRU\ YDULDEOHV EORRG DFLGEDVH VWDWXV DQG DIILQLW\ RI KDHPRJORELQ LQ $WODQWLF VDOPRQ 6DOPR VDODU /f -RXUQDO RI &RPSDUDWLYH 3K\VLRORJ\ %

PAGE 127

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f LQ WKH $PHULFDQ VRXWKZHVW 3DJHV LQ 51DLPDQ DQG '/ 6ROW] HGLWRUV )LVKHV LQ 1RUWK $PHULFDQ GHVHUWV -RKQ :LOH\ DQG 6RQV 1HZ
PAGE 128

1RUGOLH )* 7KH LQIOXHQFH RI HQYLURQPHQWDO VDOLQLW\ RQ UHVSLUDWRU\ R[\JHQ GHPDQGV LQ WKH HXU\KDOLQH WHOHRVW $PEDVVLV LQWHUUXSWD %OHHNHU &RPSDUDWLYH %LRFKHPLVWU\n DQG 3K\VLRORJ\ $ 1RUGOLH )* 2VPRWLF UHJXODWLRQ LQ WKH VKHHSVKHDG PLQQRZ &\SULQRGRQ YDULHJDWXV /DFSHGH -RXUQDO RI )LVK %LRORJ\ 1RUGOLH )* 3ODVPD RVPRWLF 1D DQG &On UHJXODWLRQ XQGHU HXU\KDOLQH FRQGLWLRQV LQ &\SULQRGRQ YDULHJDWXV /DFSHGH &RPSDUDWLYH %LRFKHPLVWU\ DQG 3KYVLRORJ\ $ 1RUGOLH )* :$ :DKO ,, %LQHOOR DQG '& +DQH\ %RG\ ZDWHU FRQWHQW LQ WKH VKHHSVKHDG PLQQRZ &\SULQRGRQ YDULHJDWXV /DFSHGH RYHU D ZLGH UDQJH RI DPELHQW VDOLQLWLHV -RXUQDO RI )LVK %LRORJ\ 1RUGOLH )* DQG 6:DOVK $GDSWLYH UDGLDWLRQ LQ RVPRWLF UHJXODWRU\ SDWWHUQV DPRQJ WKUHH VSHFLHV RI F\SULQRGRQWLGV 7HOHRVWHL $WKHULQRPRUSKDf 3K\VLRORJLFDO =RRORJ\ 1RUGOLH )* 6:DOVK '& +DQH\ DQG 7) 1RUGOLH 7KH LQIOXHQFH RI DPELHQW VDOLQLW\ RQ URXWLQH PHWDEROLVP LQ WKH WHOHRVW &\SULQRGRQ YDULHJDWXV /DFSGF -RXUQDO RI )LVK %LRORJ\ 2GXP +7 DQG '. &DOGZHOO )LVK UHVSLUDWLRQ LQ WKH QDWXUDO R[\JHQ JUDGLHQW RI DQ DQDHURELF VSULQJ LQ )ORULGD &RSHLD 2LNDZD 6 < ,WD]DZD DQG 0 *RWRK 2QWRJHQHWLF FKDQJH LQ WKH UHODWLRQVKLS EHWZHHQ PHWDEROLF UDWH DQG ERG\ PDVV LQ D VHD EUHDP 3DJUXV PDMRU 7HPPLQFN t 6FKOHJHOf -RXUQDO RI )LVK %LRORJ\ 2WW 0( 1 +HLVOHU DQG *5 8OWVFK $ UHHYDOXDWLRQ RI WKH UHODWLRQVKLS EHWZHHQ WHPSHUDWXUH DQG WKH FULWLFDO R[\JHQ WHQVLRQ LQ IUHVKZDWHU ILVKHV &RPSDUDWLYH %LRFKHPLVWU\ DQG 3K\VLRORJ\ $ 3DODFLRV &$0 DQG /* 5RVV 7KH HIIHFWV RI WHPSHUDWXUH ERG\ ZHLJKW DQG K\SR[LD RQ WKH R[\JHQ FRQVXPSWLRQ RI WKH 0H[LFDQ PRMDUUD &LFKODVRPD XURSKWKDOPXV *LLQWKHUf $TXDFXOWXUH DQG )LVKHULHV 0DQDJHPHQW 3DUHQWL /5 $ SK\ORJHQHWLF DQG ELRJHRJUDSKLF DQDO\VLV RI &\SULQRGRQWLIRUP ILVKHV 7HOHRVWHL $WKHULQRPRUSKDf %XOOHWLQ RI WKH $PHULFDQ 0XVHXP RI 1DWXUDO +LVWRU\ 3DUNHU $ DQG .RPILHOG 0ROHFXODU SHUVSHFWLYH RQ HYROXWLRQ DQG ]RRJHRJUDSK\ RI &\SULQRGRQWLG NLOOLILVKHV 7HOHRVWHL $WKHULQRPRUSKDf &RSHLD 3DUYDWKFVZDUDUDR 9 $GDSWDWLRQ WR RVPRWLF VWUHVV LQ ILVKHV ,QGLDQ %LRORJLVW 3FOVWFU % &5 %ULGJHV DQG 0. *ULFVKDEFU D 5HVSLUDWRU\ DGDSWDWLRQV RI WKH EXUURZLQJ PDULQH WHOHRVW /XQLSHQXV ODPSUHWDHIRUPLV :DOEDXPf ,, 0HWDEROLF DGDSWDWLRQV -RXUQDO RI ([SHULPHQWDO 0DULQH %LRORJ\ DQG (FRORJ\

PAGE 129

3HOVWHU % &5 %ULGJHV $& 7D\ORU 6 0RUULV DQG 5-$ $WNLQVRQ E 5HVSLUDWRUff DGDSWDWLRQV RI WKH EXUURZLQJ PDULQH WHOHRVW /XPSHQXV ODPSUHWDHIRUPLV :DOEDXPf 2W DQG &2f WUDQVSRUW DFLGEDVH EDODQFH D FRPSDULVRQ ZLWK &HSROD UXEHVFHQV / -RXUQDO RI ([SHULPHQWDO 0DULQH %LRORJ\ DQG (FRORJ\ 3HUU\ 6( DQG 0F'RQDOG *DV H[FKDQJH 3DJHV LQ '+ (YDQV HGLWRU 7KH SK\VLRORJ\ RI ILVKHV &5& 3UHVV %RFD 5DWRQ )/ S 3HWHUVRQ 06 +\SR[LDLQGXFHG SK\VLRORJLFDO FKDQJHV LQ WZR PDQJURYH VZDPS ILVKHV VKHHSVKHDG PLQQRZ &\SULQRGRQ YDULHJDWXV /DFSHGH DQG VDLOILQ PROO\ 3RHFLOLD ODWLSLQQD /HVXHXUf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
PAGE 130

5HQDXG 0/ $QQRWDWHG ELEOLRJUDSK\ RQ K\SR[LD DQG LWV HIIHFWV RQ PDULQH OLIH ZLWK HPSKDVLV RQ WKH *XOI RI 0H[LFR 86 'HSDUWPHQW RI &RPPHUFH 1DWLRQDO 2FHDQLF DQG $WPRVSKHULF $GPLQLVWUDWLRQ 1DWLRQDO 0DULQH )LVKHULHV 6HUYLFH 12$$ 7HFKQLFDO 5HSRUW 10)6 5RVV % DQG /* 5RVV 7KH R[\JHQ UHTXLUHPHQWV RI 2UHRFKURPLV QLORWLFXV XQGHU DGYHUVH FRQGLWLRQV 3DJHV LQ / )LVKHOVRQ DQG =
PAGE 131

6SDDUJDUHQ '+ $ VWXG\ RQ WKH DGDSWDWLRQ RI PDULQH RUJDQLVPV WR FKDQJLQJ VDOLQLWLHV ZLWK VSHFLDO UHIHUHQFH WR WKH VKRUH FUDE &DUFLQXV PDHQDV /f &RPSDUDWLYH %LRFKHPLVWU\ DQG 3K\VLRORJ\ $ 6WRXW -3 7KH HFRORJ\ RI LUUHJXODUO\ IORRGHG VDOW PDUVKHV RI WKH QRUWKHDVWHUQ *XOI RI 0H[LFR D FRPPXQLW\ SURILOH 8QLWHG 6WDWHV )LVK DQG :LOGOLIH 6HUYLFH %LRORJLFDO 5HSRUW f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

PAGE 132

7UXHVGDOH *$ $/ 'RZQLQJ DQG *) /RZGHQ 7KH VROXELOLW\ RI R[\JHQ LQ SXUH ZDWHU DQG VHD ZDWHU -RXUQDO RI $SSOLHG &KHPLVWU\ 7XUQHU %DQG 5. /LX ([WHQVLYH LQWHUVSHFLILF JHQHWLF FRPSDWDELOLW\ LQ WKH QHZ ZRUOG NLOOLILVK JHQXV &\SULQRGRQ &RSHLD 8OWVFK *5 + %RVFKXQJ DQG 05RVV 0HWDEROLVP FULWLFDO R[\JHQ WHQVLRQ DQG KDELWDW VHOHFWLRQ LQ GDUWHUV (WKHRVWRPDf (FRORJ\ 9HQWUHOOD 9 ) 7URPEHWWL $ 3DJOLDUDQL 7ULJDUL 0 3LULQL DQG $5 %RUJDWWL 6DOLQLW\ GHSHQGHQFH RI WKH RXDEDLQLQVHQVLWLYH 0JfGHSHQGHQW 1D $73DVH LQ JLOOV RI UDLQERZ WURXW 2QFRUK\QFKXV P\NLVV :DOEDXPf DGDSWHG WR IUHVK DQG EUDFNLVK ZDWHU &RPSDUDWLYH %LRFKHPLVWU\ DQG 3K\VLRORJ\ %O 9HUKH\HQ ( 5 %OXVW DQG : 'HFOHLU 0HWDEROLF UDWH K\SR[LD WROHUDQFH DQG DTXDWLF VXUIDFH UHVSLUDWLRQ RI VRPH ODFXVWULQH DQG ULYHULQH $IULFDQ ILVKHV 3LVFHV &LFKOLGDHf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f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

PAGE 133

:HOOV 50* 0' $VKE\ 6'XQFDQ DQG -$ 0DF'RQDOG &RPSDUDWLYH VWXG\ RI WKH HU\WKURF\WHV DQG KDHPRJORELQV LQ QRWRWKHQLLG ILVKHV IURP $QWDUFWLFD -RXUQDO RI )LVK %LRORJ\ :HOOV 50* *& *ULJJ /$ %HDUG DQG 6XPPHUV +\SR[LF UHVSRQVHV LQ D ILVK IURP D VWDEOH HQYLURQPHQW EORRG R[\JHQ WUDQVSRUW LQ WKH DQWDUFWLF ILVK 3DJRWKHQLD ERUFKJUHYLQNL -RXUQDO RI ([SHULPHQWDO %LRORJ\ :HOOV 50* DQG 5( :HEHU ,V WKHUH DQ RSWLPDO KDHPDWRFULW IRU UDLQERZ WURXW 2QFRUK\QFKXV P\NLVV :DOEDXPf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
PAGE 134

%,2*5$3+,&$/ 6.(7&+ 'HQQLV &KDUOHV +DQH\ ZDV ERP 0DUFK LQ ,QJOHZRRG &DOLIRUQLD WR &KDUOHV DQG -HDQQH +DQH\ +H JUHZ XS LQ 6RXWKHUQ &DOLIRUQLD ZLWK KLV SDUHQWV DQG ROGHU EURWKHU 6FRWW JUDGXDWLQJ IURP &KDWVZRUWK +LJK 6FKRRO LQ 'HQQLV EHJDQ KLV FDUHHU LQ WKH ELRORJLFDO VFLHQFHV DV DQ XQGHUJUDGXDWH DW WKH 8QLYHUVLW\ RI &DOLIRUQLD 6DQ 'LHJR 8&6'f :KLOH DW 8&6' 'HQQLV KDG KLV ILUVW WUXH H[SRVXUH WR WKH MR\V DQG SLWIDOOVf RI UHVHDUFK DQG WHDFKLQJ +H JUDGXDWHG LQ ZLWK D %$ LQ ELRORJ\ VSHFLDOL]DWLRQ LQ DQLPDO SK\VLRORJ\f DQG PRYHG RQ WR WKH JUDGXDWH SURJUDP DW 2UHJRQ 6WDWH 8QLYHUVLW\ 268f 'HQQLV VSHQW WZR \HDUV DW 268 ZKHUH KH ZDV LQWURGXFHG WR WKH VWXG\ RI ILVK SK\VLRORJ\ )RU KLV PDVWHUnV WKHVLV KH H[DPLQHG WKH SK\VLRORJLFDO DQG KHPDWRORJLFDO HIIHFWV RI HU\WKURF\WLF QHFURVLV YLUXV RQ FKXP VDOPRQ 2QFRUK\QFKXV NHWDf 'HQQLV FRPSOHWHG KLV 06 LQ ELRORJLFDO VFLHQFH LQ IROORZLQJ ZKLFK KH PRYHG DFURVV WKH FRXQWU\ WR EHJLQ D GRFWRUDO SURJUDP DW WKH 8QLYHUVLW\ RI )ORULGD 8)f 6LQFH 'HQQLV KDV VLPXOWDQHRXVO\ ZRUNHG RQ KLV GLVVHUWDWLRQ DQG IRU WKH 'HSDUWPHQW RI WKH ,QWHULRU DV D %LRORJLFDO 7HFKQLFLDQ 'HQQLV FRPSOHWHG KLV GLVVHUWDWLRQ DQG JUDGXDWHG IURP 8) LQ

PAGE 135

, FHUWLI\ WKDW KDYH UHDG WKLV VWXG\ DQG WKDW LQ P\ RSLQLRQ LW FRQIRUPV WR DFFHSWDEOH VWDQGDUGV RI VFKRODUO\ SUHVHQWDWLRQ DQG LV IXOO\ DGHTXDWH LQ VFRSH DQG TXDOLW\ DV D GLVVHUWDWLRQ IRU WKH GHJUHH RI 'RFWRU RI 3KLORVRSK\ )UDQNf“RIFOOH &KDLU 3URIHVVRU RI =RRORJ\ FHUWLI\ WKDW KDYH UHDG WKLV VWXG\ DQG WKDW LQ P\ RSLQLRQ LW FRQIRUPV WR DFFHSWDEOH VWDQGDUGV RI VFKRODUO\ SUHVHQWDWLRQ DQG LV IXOO\ DGHTXDWH LQ VFRSH DQG TXDOLW\ DV D GLVVHUWDWLRQ IRU WKH GHJUHH RI 'RFWRU RI 3KLORVRSK\ ? E\ /LOL nZKLWHn ‘+DUYH\ /LOL 3URIHVVRU R nZKLWH =RRORJ\ FHUWLI\ WKDW KDYH UHDG WKLV VWXG\ DQG WKDW LQ P\ RSLQ RQ LW FRQIRUPV WR DFFHSWDEOH VWDQGDUGV RI VFKRODUO\ SUHVHQWDWLRQ DQG LV IXOO\ DGHTXDWH LQ VFRSH DQG TXDOLW\ DV D GLVVHUWDWLRQ IRU WKH GHJUHH RI 'RFWRU RI 3KLORVRSK\ %UDQA0[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

PAGE 136

, FHUWLI\ WKDW KDYH UHDG WKLV VWXG\ DQG WKDW LQ P\ RSLQLRQ LW FRQIRUPV WR DFFHSWDEOH VWDQGDUGV RI VFKRODUO\ SUHVHQWDWLRQ DQG LV IXOO\ DGHTXDWH LQ VFRSH DQG TXDOLW\ DV D GLVVHUWDWLRQ IRU WKH GHJUHH RI 'RFWRU RI 3KLORVRSK\ 7QRPDV &ULVPDQ 3URIHVVRU RI (QYLURQPHQWDO (QJLQHHULQJ 6FLHQFHV 7KLV GLVVHUWDWLRQ ZDV VXEPLWWHG WR WKH *UDGXDWH )DFXOW\ RI WKH 'HSDUWPHQW RI =RRORJ\ LQ WKH &ROOHJH RI /LEHUDO $UWV DQG 6FLHQFHV DQG WR WKH *UDGXDWH 6FKRRO DQG ZDV DFFHSWHG DV SDUWLDO IXOILOOPHQW RI WKH UHTXLUHPHQWV IRU WKH GHJUHH RI 'RFWRU RI 3KLORVRSK\ 'HFHPEHU 'HDQ *UDGXDWH 6FKRRO

PAGE 137

K8WI 81,9(56,7< 2) )/25,'$


xml version 1.0 encoding UTF-8
REPORT xmlns http:www.fcla.edudlsmddaitss xmlns:xsi http:www.w3.org2001XMLSchema-instance xsi:schemaLocation http:www.fcla.edudlsmddaitssdaitssReport.xsd
INGEST IEID ELIYC40N1_1XLF0E INGEST_TIME 2012-02-20T23:02:44Z PACKAGE AA00009024_00001
AGREEMENT_INFO ACCOUNT UF PROJECT UFDC
FILES