• TABLE OF CONTENTS
HIDE
 Title Page
 Certifications
 Copyright
 Acknowledgement
 Table of Contents
 List of Tables
 List of Figures
 Abstract
 Introduction
 Methods and materials
 Results
 Discussion
 Conclusion
 Appendix A: Site records for mussel...
 Appendix B: Site records for fish...
 Reference
 Biographical sketch






Group Title: Reproductive biology of Elliptoideus sloatianus, Lampsilis subangulata, and Pleurobema pyriforme (Bivalvia: Unionidae)
Title: Reproductive biology of Elliptoideus sloatianus, Lampsilis subangulata, and Pleurobema pyriforme (Bivalvia Unionidae)
CITATION PAGE IMAGE ZOOMABLE
Full Citation
STANDARD VIEW MARC VIEW
Permanent Link: http://ufdc.ufl.edu/UF00073813/00001
 Material Information
Title: Reproductive biology of Elliptoideus sloatianus, Lampsilis subangulata, and Pleurobema pyriforme (Bivalvia Unionidae)
Physical Description: ix, 72 leaves : ill. ; 29 cm.
Language: English
Creator: O'Brien, Christine A., 1968-
Publication Date: 1997
 Subjects
Subject: Mussels -- Physiology   ( lcsh )
Unionoida -- Physiology   ( lcsh )
Mussels -- Reproduction   ( lcsh )
Unionoida -- Reproduction   ( lcsh )
Wildlife Ecology and Conservation thesis, M.S   ( lcsh )
Dissertations, Academic -- Wildlife Ecology and Conservation -- UF   ( lcsh )
Genre: bibliography   ( marcgt )
non-fiction   ( marcgt )
 Notes
Thesis: Thesis (M.S.)--University of Florida, 1997.
Bibliography: Includes bibliographical references (leaves 65-69).
Statement of Responsibility: by Christine A. O'Brien.
General Note: Typescript.
General Note: Vita.
Funding: This collection includes items related to Florida’s environments, ecosystems, and species. It includes the subcollections of Florida Cooperative Fish and Wildlife Research Unit project documents, the Sea Grant technical series, the Florida Geological Survey series, the Coastal Engineering Department series, the Howard T. Odum Center for Wetland technical reports, and other entities devoted to the study and preservation of Florida's natural resources.
 Record Information
Bibliographic ID: UF00073813
Volume ID: VID00001
Source Institution: University of Florida
Holding Location: University of Florida
Rights Management: All rights reserved, Board of Trustees of the University of Florida
Resource Identifier: aleph - 002266980
oclc - 37210669
notis - ALL9967

Table of Contents
    Title Page
        Title page
    Certifications
        i
    Copyright
        ii
    Acknowledgement
        iii
    Table of Contents
        iv
        v
    List of Tables
        vi
    List of Figures
        vii
    Abstract
        viii
        ix
    Introduction
        Page 1
        Historical background
            Page 1
            Page 2
        Freshwater mussel biology
            Page 3
            Page 4
            Page 5
            Page 6
            Page 7
            Page 8
            Page 9
            Page 10
            Page 11
    Methods and materials
        Page 12
        Study site
            Page 12
            Page 13
            Page 14
        Reproductive biology
            Page 15
            Page 16
            Page 17
            Page 18
            Page 19
            Page 20
            Page 21
            Page 22
        Development of immunity
            Page 23
        Glochidia reaction to nonindigenous fish
            Page 23
        Glochidial morphology
            Page 24
    Results
        Page 25
        Reproductive biology
            Page 25
            Page 26
            Page 27
            Page 28
            Page 29
            Page 30
            Page 31
            Page 32
            Page 33
            Page 34
            Page 35
            Page 36
            Page 37
        Development of immunity
            Page 38
            Page 39
            Page 40
        Glochidia reaction to nonindigenous fish
            Page 41
        Glochidia morphology
            Page 41
            Page 42
            Page 43
            Page 44
            Page 45
            Page 46
            Page 47
            Page 48
            Page 49
    Discussion
        Page 50
        Reproductive biology
            Page 50
            Page 51
            Page 52
            Page 53
        Development of immunity
            Page 54
        Glochidia reaction to nonindigenous fish
            Page 55
        Glochidia morphology
            Page 56
        Conservation issues
            Page 57
            Page 58
            Page 59
    Conclusion
        Page 60
        Page 61
        Page 62
    Appendix A: Site records for mussel collection sites
        Page 63
        Page 64
    Appendix B: Site records for fish collection sites
        Page 65
        Page 66
    Reference
        Page 67
        Page 68
        Page 69
        Page 70
        Page 71
    Biographical sketch
        Page 72
Full Text













REPRODUCTIVE BIOLOGY OF ELLIPTOIDEUS SLOATIANUS, LAMPSILIS
SUBANGULATA, MEDIONIDUS PENICILLATUS, AND PLEUROBEMA
PYRIFORME
(BIVALVIA: UNIONIDAE)














By

Christine A. O'Brien


A THESIS PRESENTED TO THE GRADUATE SCHOOL
OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT
OF THE REQUIREMENTS FOR THE DEGREE OF
MASTER OF SCIENCE

UNIVERSITY OF FLORIDA


1997









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 thesis for the degree of Master of science.


/Georg/ Tanner, Chair
Associate Professor of
Wildlife Ecology and
Conservation


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 thesis for the degree of Master of Science.


Oi--
/J es Williams, Cochair
Adunct Professor of
ldlife Ecology and
Conservation


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 thesis for the degree of Master of Science.


Loukas Arvanitis
Professor of Forest
Resources and Conservation


This thesis was submitted to the Graduate Faculty of
the College of Agriculture and to the Graduate School and
was accepted as partial fulfillment of the requirements for
the degree of Master of Science.

May, 1997 ____
Dea College of
Agriculture


Dean, Graduate School



























Copyright 1997

by

Christine A. O'Brien















ACKNOWLEDGMENTS


The funding for this project was provided by the U. S.

Fish and Wildlife Service in Jacksonville, Florida. I would

like to thank Jayne Brim Box for introducing me to the

fascinating world of freshwater mussels. Jayne also

provided me with much needed guidance throughout my thesis

work, both in the field and laboratory. Jayne was always

willing to answer any questions I had about my work. I

would like to thank Shane Ruessler for his enthusiastic

assistance in both the field and laboratory. A special

thanks to Noel Burkhead and Howard Jelks for providing me

with information about fish biology and editorial comments.

I would also like to thank my committee members for

supporting my efforts during my work towards my Master of

Science degree.

Finally, I would like to thank my family for providing

me with love and support throughout my studies.


iii










TABLE OF CONTENTS


ACKNOWLEDGMENTS . .

LIST OF TABLES . .

LIST OF FIGURES . .


iii


. . vi

. . vii


ABSTRACT


viii


1 INTRODUCTION . . . .

Historical Background . .
Freshwater Mussel Biology . .

2 METHODS AND MATERIALS . . .

Study Site . . . .
Target Mussel Species . . .
Reproductive Biology . . .
Development of Immunity . .
Glochidia Reaction to Nonindigenous Fish
Glochidia Morphology . . .

3 Results . . . . .

Reproductive Biology . . .
Development of Immunity . .
Glochidia Reaction to Nonindigenous Fish
Glochidia morphology . . .


3
. . 3

. . 12


. . 25


S 25
S 38
S 41
S 41


4 Discussion


. 50


Reproductive Biology . . .
Development of Immunity . .
Glochidia Reaction to Nonindigenous Fish
Glochidia Morphology . . .
Conservation Issues . . .

5 Conclusion . . . .

APPENDICES

A-1 SITE RECORDS FOR MUSSEL COLLECTION SITES

B-l SITE RECORDS FOR FISH COLLECTION SITES


. . 65













LIST OF REFERENCES

BIOGRAPHICAL SKETCH


. 72















LIST OF TABLES


Tables page

2-1. List of fish names . . .. .. 19

3-1. Months when the mussels were found gravid . 26

3-2. Host fish and non-host fish for E. sloatianus 27

3-3. Percent fish producing E. sloatianus juveniles .29

3-4. Dates when superconglutinantes are released .30

3-5. Host fish and non-host fish for L. subangulata .32

3-6. Percent fish producing L. subangulata juveniles .33

3-7. Host fish and non-host fish for M. penicillatus .35

3-8. Percent fish producing M. penicillatus juveniles .37

3-9. Host fish and non-host fish for P. pyriforme .39

3-10. Percent fish producing P. pyriforme juveniles 40

3-11. Number of L. subangulata after two infestations .42

3-12. Nonindigenous fish reaction to glochidia . 43
















LIST OF FIGURES


Figures page

1-1. Freshwater mussel life cycle . . 5

1-2. Superconglutinate . . . . 7

3-1. Elliptoideus sloatianus glochidia morphology 44

3-2. Lampsilis subangulata glochidia morphology .46

3-3. Medionidus penicillatus glochidia morphology .47

3-4. Pleurobema pyriforme glochidia morphology . 48


vii















Abstract of Thesis Presented to the Graduate School
of the University of Florida in Partial Fulfillment of the
Requirements for the Degree of Master of Science


REPRODUCTIVE BIOLOGY OF ELLIPTOIDEUS SLOATIANUS, LAMPSILIS
SUBANGULATA, MEDIONIDUS PENICILLATUS, AND PLEUROBEMA
PYRIFORME
(BIVALVIA: UNIONIDAE)

By

Christine A. O'Brien

May 1997

Chairperson: Dr. George Tanner
Major Department: Wildlife Ecology and Conservation

A study on the reproductive biology of Elliptoideus

sloatianus, Lampsilis subangulata, Medionidus penicillatus,

and Pleurobama pyriforme was conducted from May 1995 to

January 1997. The mussels used in the study were collected

in the Apalachicola, Chattahoochee, Flint, and Ochlockonee

river drainages in Alabama, Georgia, and Florida. Periods of

gravidity were determined by inspecting female mussels

monthly; host fish were determined by laboratory infections.

A test was conducted to determine if host fish developed an

immunity to glochidia after multiple infestations occurred.

An experiment was also conducted to determine if unionid

glochidia will transform on a nonindigenous fish. Finally,

the glochidia from each mussel species were photographed

viii









using a scanning electron microscope. Primary host fish

were not identified for Elliptoideus sloatianus.

Elliptoideus sloatianus is a tachytictic breeder. Lampsilis

subangulata is a bradytictic breeder. Primary host fish

were identified as Micropterus punctulatus and M. salmoides.

Medionidus penicillatus is a bradytictic breeder. Primary

host fish were identified as Etheostoma edwini and Percina

nigrofasciata. Pleurobama pyriforme is a tachytictic

breeder. The primary host fish was identified as

Pteronotropis hypselopterus. After infesting M. salmoides

twice with L. subangulata glochidia, a t-test indicated that

there was a significant reduction in the number of juveniles

produced by M. salmoides after the second infection.

Poecilia reticulata, a nonindigenous fish, successfully

transformed glochidia from all four mussel species.

However, the number of Poecilia retuculata producing

juvenile mussels varied for each species of mussel.















CHAPTER 1
INTRODUCTION


Historical Background



Historically, North America was known for its rich

mussel fauna with about 300 species recognized (Turgeon et

al. 1988). Since the early 1900s, 6% of the mussel fauna

has become extinct and 21% are federally listed as

endangered or threatened (Neves 1992; Williams et al. 1992).

The reasons for this decline are not well understood.

However, loss of suitable habitat (caused by increased

siltation and dredging), introductions of nonindigenous

species, pollution, and commercial exploitation are thought

to have a negative impact on mussel populations (Ortmann

1909; Fuller 1974; Williams et al. 1992). In addition,

mussels are obligate parasites on fish, and recent declines

in the North American fish fauna may also have negative

effects on mussel populations (Allen and Flecker 1993). In

fact, aquatic species of North America have an imperilment

rate three to eight times greater than the imperilment rate

of avian and mammalian fauna (Master 1990).

In 1994, seven species of mussels from the eastern Gulf

of Mexico drainage were proposed for federal listing as an

endangered or threatened species under the Endangered















CHAPTER 1
INTRODUCTION


Historical Background



Historically, North America was known for its rich

mussel fauna with about 300 species recognized (Turgeon et

al. 1988). Since the early 1900s, 6% of the mussel fauna

has become extinct and 21% are federally listed as

endangered or threatened (Neves 1992; Williams et al. 1992).

The reasons for this decline are not well understood.

However, loss of suitable habitat (caused by increased

siltation and dredging), introductions of nonindigenous

species, pollution, and commercial exploitation are thought

to have a negative impact on mussel populations (Ortmann

1909; Fuller 1974; Williams et al. 1992). In addition,

mussels are obligate parasites on fish, and recent declines

in the North American fish fauna may also have negative

effects on mussel populations (Allen and Flecker 1993). In

fact, aquatic species of North America have an imperilment

rate three to eight times greater than the imperilment rate

of avian and mammalian fauna (Master 1990).

In 1994, seven species of mussels from the eastern Gulf

of Mexico drainage were proposed for federal listing as an

endangered or threatened species under the Endangered









2

Species Act of 1973. The seven species included the fat

threeridge (Amblema neislerii), Chipola slabshell (Elliptio

chipolaensis), purple bankclimber (Elliptoideus sloatianus),

shinyrayed pocketbook (Lampsilis subangulata), Gulf

moccasinshell (Medionidus penicillatus), Ochlockonee

moccasinshell (Medionidus simpsonianus), and oval pigtoe

(Pleurobema pyriforme)(Federal Register 1994).

Historically, mussel shell was used to manufacture

buttons. In 1898 over 50 companies were in production along

the Mississippi River, producing over 30 million buttons in

only a few years. The high demand for shell products in the

late 1800s reduced mussel populations and prompted

conservation efforts to be implemented (Jones 1950). In

1894, the United States Bureau of Fisheries encouraged

studies in the artificial propagation of mussels to help

alleviate the high demand on natural mussel beds (Coker

1919). Despite these early efforts to conserve mussel

populations, their numbers continued to decline.

After the decline of the button industry, mussel

harvesting was greatly reduced for many years. However, in

the 1950s mussel harvesting increased as the pearl industry

began to grow. Today, mussel shell is used to seed pearl

oysters in China and Japan. In 1991, the pearl industry

encouraged the collection of 9,000 tons of mussel shell in

North America. In 1992 the harvest dropped to only 4,500

tons (Williams et al. 1992).









3

Freshwater Mussel Biology



Freshwater mussels are an important component of the

aquatic ecosystem. Freshwater mussels remove toxins, silt,

and other organic material from the water column, and

provide food for wildlife (Stansbery and Stein 1971;

Williams et al. 1992). Freshwater mussels are also good

indicators of water quality because they are long lived,

widely distributed, sessile organisms which are sensitive to

pollution (Stansbery and Stein 1971).

Freshwater mussels are unique among bivalves in that

most require a host fish to complete their life cycle.

Unlike male and female marine bivalves which release sperm

and eggs into the water column, fertilization of freshwater

mussels takes place within broodchambers in the gills of the

female (Jirka and Neves 1992). A broodchamber filled with

eggs and or mature glochidia is usually identified by its

swollen appearance (Coker 1919). The female mussel

carries the fertilized eggs in the gills until they develop

into a parasitic stage the called glochidia stage.

Mussels have been categorized into two groups based on

their embryo incubation period (Lefevre and Curtis 1912;

Waller and Holland-Bartels 1988). The first group,

tachytictic (short-term) breeders, fertilize eggs in the

spring and release glochidia in the fall. The second group,

bradytictic (long-term) breeders, fertilize eggs in late









4

summer and release glochidia in the following spring or

early summer. When fully developed, the female mussels

release the glochidia into the water column where they must

come into contact with the proper host fish (Figure 1-1)

Once attached to the host fish, the glochidia metamorphose

and drop to the substrate to become free living juveniles.

The mussel/fish relationship is often species-specific

(Lefevre and Curtis 1912). Only certain species of fish can

serve as suitable hosts for a particular mussel species. In

some mussels (i.e., Strophitus sp. and Uterbackia sp.)

glochidia do not require a host fish for transformation

(Lefevre and Curtis 1912).

The attachment of glochidia to a proper host fish is

often difficult. Female mussels increase the probability of

successful glochidia/host fish interactions by producing

thousands of glochidia (Jansen 1990). Female mussels can

(depending on size and species) produce anywhere from 2,000

(Kondo 1984) to as many as 3,000,000 glochidia (Coker et al.

1921). Some mussel species attempt to increase the chance

of glochidia contacting a proper host fish by releasing

glochidia into the water column when light sensitive spots

on the mantle are stimulated by the shadow of a passing fish

(Kraemer 1970; Jansen 1990). Other mussels lure host fish

by extending brightly colored portions of their mantles to

mimic prey (Coker et al. 1921; Kraemer 1970).





















Glochidia attach to
gill filaments or fins


Sperm


- Juvenile mussel


Glochidia


Female Male


Figure 1-1. Freshwater mussel life cycle.









6

The use of lures to increase a mussel's chance of

infesting a host fish is not uncommon. In fact, several

mussel species in the subfamily Lampsiline have developed

lures, mainly in the form of modified mantle flaps in the

shape of prey (Kraemer 1970). The lures displayed by the

female mussels disillusions the host fish by mimicing a

consumable meal. When the host fish attempts to consume a

mimic lure, the female mussel releases glochidia into the

water column where attachment to the host fish can occur.

Perhaps one of the most amazing forms of a fish mimic is

that of a superconglutinate (Haag et al. 1995). A

superconglutinate holds the entire reproductive efforts for

the year in a package that mimics a small fish. The

superconglutinate consists of a mucus strand attached to a

modified conglutinate (Figure 1-2).

A conglutinate is a packet of glochidia, or eggs, that

takes the shape of water tubes of the gills in female

mussels. The size and shape of a conglutinate depends on

the species of mussel. The modified conglutinate is

comprised of single conglutinates that have been fused

together by the mucus strand. Most mussel species release

single conglutinates rather than fusing the conglutinates

(Coker et al. 1921). The length of the modified

conglutinate segment of the superconglutinate ranges from

three to five centimeters. The modified conglutinate not

only resembles a small fish in size and shape but in













































C



0)
C
0

Cs)



C,)


=I
-5,
0
0
V













U,

U)
C.


04
r0

c,








.,
Cl






0





4-J









4-
COr







*- C
*^
*-1
CO 4









8

coloration as well. The modified conglutinate has

pigmentation in the form of a lateral band and eye spot.

During the production of the superconglutinate the water

currents move the fish mimic in motions that are similar to

a small fish.

The mimic fish structure (modified conglutinate) is

attached to a mucus thread which can be produced to lengths

up to one meter. After the complete production of a

superconglutinate, which can take about four hours, the

female mussel discontinues the production of mucus and the

superconglutinate breaks free from her excurrent siphon.

After detaching from the female, the superconglutinate

drifts passively in the stream current. The fate of the

superconglutinate now depends on the chance that the current

will wrap the superconglutinate around a rock, branch, or

any structure in the stream where it will continue to mimic

prey for a piscivorous host fish. When a fish attempts to

consume the superconglutinate thousands of the glochidia

within the modified conglutinate are released and may attach

to the fish and later fall off as juvenile mussels.

Glochidia failing to come into contact with a suitable

host will drift through the water column, surviving for only

a few days (Jansen 1990). When the glochidia come in

contact with a suitable host, attachment to the gill and/or

fin region will occur and the glochidium will become fully

encysted within 24 to 36 hours (Coker et al. 1921). The









9

encysting stage of the glochidia on the fish is a fish's

response to the puncture caused by the clamping of a

glochidium. Glochidia attaching to a non-host fish will

slough off soon after attachment occurs. Properly encysted

glochidia will metamorphose, with little change in size

(Coker et al. 1921), into free-living juveniles, displaying

abductor muscles, gill buds, and a ciliated foot with

protractor and retractor muscles (Karna and Millemann 1978).

The time required for complete metamorphosis of

glochidia is dependent in part on temperature and mussel

species (Jones 1950). One species of mussel, Quadrula

intermedia, takes 24 to 29 days to metamorphose in water at

190 C (Yeager and Saylor 1989), while Lampsilis radiata

takes up to 98 days at 150 C for metamorphosis to occur

(Tedla and Fernando 1969). Another mussel, Simpsoniconcha

ambigua, produces glochidia that metamorphose over the cold

winter months and requires up to seven months to complete

transformation (Howard 1951).

The reproductive biology of many freshwater mussels

remains incomplete (Jansen 1990). For example, host fish

for only 25% of the 300 mussel species in North America have

been identified (Watters 1992). Unfortunately, fewer than

25% of the host fish have been identified for the mussels of

the Gulf of Mexico drainages.

To enhance conservation efforts, fish hosts must be

identified for all mussel species with an emphasis on those









10

species proposed for endangered and threatened status.

There are two methods used to identify host fish for

freshwater mussels. The first method identifies host fish

via laboratory infestations. In the laboratory, known

species of glochidia are infected on known fish species.

The fish are then held until transformed gi chidia drop from

the fish. The second method identifies host fish via

identifying encysted glochidia, to the species level, on

fish collected from streams. A fish species that produced

juvenile mussels in the laboratory and was also found with

encysted glochidia, from the same mussel species, in a

stream is a confirmed primary host fish (Trdan and Hoeh

1982; Neves et al. 1989).

The identification of glochidia to the species level

can be difficult. In some cases streams have few species of

mussels, and the size and shape of the glochidia are

diagnostic (Neves et al. 1989). However, streams that

support a diversity of mussel species, such as those in the

southeastern United States, can make glochidia

identification to the species level almost impossible

without the aid of scanning electron microscopy (Burch 1975;

Hoggarth 1988; Waller et al. 1988).

Once the host fish have been identified for a mussel

species, more information can be focused on the status of

the host fish population. For example, a mussel species may

be forced to utilize the same host fish repeatedly in









11

streams where there are low numbers of host fish available.

Researchers have discovered that some fish develop an

immunity to glochidia when infected repeatedly (Coker et al.

1921; Karna and Millemann 1978; Luo 1993). Since most

mussels are obligate parasites on fish, it is important that

conservation efforts are focused on both mussel and fish

populations, so that all phases of the life cycle are

protected.

The five main objectives of this study are as follows:

1) determine when Elliptoideus sloatianus, Lampsilis

subangulata, Medionidus penicillatus, and Pleurobema

pyriforme produce mature glochidia, 2) identify the primary

host fish for the four target mussel species, 3) test for

the development of an immune response when a fish is

infected twice with the same species of glochidia, 4) test

whether a nonindigenous fish has an immunity to glochidia of

the target mussel species, and 5) describe the glochidia of

each of the target mussel species using the scanning

electron microscopy (SEM).















CHAPTER 2
MATERIALS AND METHODS

Study Site



The Apalachicola, Chattahoochee, Flint, and Ochlockonee

river drainages cover approximately 41,979 km2 of the

southeastern United States, making it one of the largest

river drainage systems in the eastern Gulf Coastal Plain

(Leitman et al. 1983). The majority of mussel species occur

in the low gradient streams of the Coastal Plain and

Piedmont regions. Most streams in the Coastal Plain region

typically have substrate of coarse sand, silty sand, and

gravel. However, the substrata of the Apalachicola and

Ochlockonee rivers consists of a coarser substrate of

cobble, boulders, and bedrock. The Apalachicola,

Chattahoochee, Flint, and Ochlockonee River basin has a rich

aquatic fauna consisting of 30 crayfish species, 20 species

of aquatic snails, 122 fish species, and 32 endemic mussel

species (Butler 1989; Couch et al. 1996).



Target Mussel Species



Elliptoideus sloatianus is a large mussel with a

maximum length of 203 mm and is usually found in sand, muddy

12















CHAPTER 2
MATERIALS AND METHODS

Study Site



The Apalachicola, Chattahoochee, Flint, and Ochlockonee

river drainages cover approximately 41,979 km2 of the

southeastern United States, making it one of the largest

river drainage systems in the eastern Gulf Coastal Plain

(Leitman et al. 1983). The majority of mussel species occur

in the low gradient streams of the Coastal Plain and

Piedmont regions. Most streams in the Coastal Plain region

typically have substrate of coarse sand, silty sand, and

gravel. However, the substrata of the Apalachicola and

Ochlockonee rivers consists of a coarser substrate of

cobble, boulders, and bedrock. The Apalachicola,

Chattahoochee, Flint, and Ochlockonee River basin has a rich

aquatic fauna consisting of 30 crayfish species, 20 species

of aquatic snails, 122 fish species, and 32 endemic mussel

species (Butler 1989; Couch et al. 1996).



Target Mussel Species



Elliptoideus sloatianus is a large mussel with a

maximum length of 203 mm and is usually found in sand, muddy

12









13

sand, and cobble in moderate current (Heard 1975).

Elliptoideus sloatianus is most commonly found in deeper

sections of flowing streams and does not tolerate impounded

waters (Williams and Butler 1994).

Elliptoideus sloatianus was historically found in the

mainstem of the Apalachicola, Chattahoochee, Chipola, Flint,

and Ochlockonee rivers. Currently, E. sloatianus exist only

in the lower unimpounded mainstem of the Flint River and

several areas along the Ochlockonee River (Williams and

Butler 1994).

Lampsilis subangulata, the shinyrayed pocketbook, is a

medium-sized mussel with a maximum length of 85 mm and is

found in sandy or silty sand substrates in moderate to slow

currents. The shell is smooth and yellow in color with dark

green rays (Clench and Turner 1956). Lampsilis subangulata

is not tolerant of stream impoundments.

Historically, Lampsilis subangulata was found in the

Flint River, the mainstem of the Chattahoochee River

(including six of its tributaries), throughout the

Apalachicola River tributaries, Chipola River, and several

sites in the Ochlockonee River system (Williams and Butler

1994).

Today, the distribution of Lampsilis subangulata has

declined to one-third of its original range. Currently,

populations occur in Uchee Creek (tributary of Chattahoochee

River), a few sites in the Flint River system, Chipola








14

River, and a few sites in the upper part of the Ochlockonee

River (Williams and Butler 1994).

Medionidus penicillatus, the Gulf moccasinshell, is a

small mussel with a maximum length of approximately 53 mm.

M. penicillatus is found in sand and gravel areas with slow

to moderate currents (Heard 1975). Medionidus penicillatus

has not been found to survive in impounded river sections.

Historically, Medionidus penicillatus was found in

tributaries and main channel of the Chattachoochee and Flint

rivers, and tributaries of the Apalachicola River (Clench

and Turner 1956). The overall distribution of M.

penicillatus has declined by 75% of its original range

(Williams and Butler 1994).

Pleurobema pyriforme, the oval pigtoe, is a medium-

sized mussel with maximum lengths of 56 mm (Clench and

Turner 1956). Pleurobema pyriforme is usually found in

substrate ranging from sand to sand/gravel usually in the

center of stream channels. Pleurobema pyriforme does not

tolerate impoundments (Williams and Butler 1994).

Pleurobema pyriforme was once found in the mainstems of

the Flint, Chattachoochee, and Apalachicola River systems

(Clench and Turner 1956). Recent surveys indicate the

current distribution of P. pyriforme encompasses only two-

thirds of its historical distribution with no current

populations documented in Alabama (Williams and Butler

1994).









15

Reproductive Biology



Mussels were collected monthly and inspected for mature

glochidia starting in May 1995 and ending in April 1997.

Female mussels found with fully developed glochidia were

considered gravid. Female mussels found with eggs or

developing eggs will be mentioned.

Some monthly collections were not conducted due to

adverse field conditions. Mussels were collected from four

streams that were known to have existing populations of

Elliptoideus sloatianus, Lampsilis subangulata, Medionidus

penicillatus, and Pleurobema pyriforme. All of the streams

from which the mussels were collected occurred in the

Apalachicola, Chattachoochee, Flint, and Ochlockonee river

drainages. The mussels were collected by hand, using

snorkeling and scuba gear.

Elliptoideus sloatianus were collected at two sites

located on the Ochlockonee River and one site on the

Apalachicola River. Elliptoideus sloatianus were collected

from September 1995 to April 1997. Lampsilis subangulata

were collected from the following three creeks in southwest

Georgia: Chickasawhatchee, Cooleewahee, and Kinchafoonee.

L. subangulata were inspected for glochidia from May 1995 to

July 1996. Medionidus penicillatus and Pleurobema pyriforme

were collected from two streams, Chickasawhatchee and

Kinchafoonee, in southwest Georgia from February 1996 to

January 1997 (Appendix A-i).









16

Mussels were opened about 0.5 cm gills were and

inspected gills for the presence of glochidia. When gravid

mussels were found, they were carefully put into plastic

bags and placed into coolers with enough ice to maintain the

stream temperature during the transport to the U. S.

Geological Surveyy (USGS) laboratory. Mussels were

separated by species into separate four-liter jars and

aerated until the glochidia were collected.

The mussel species used in this experiment are proposed

for protection under the Endangered Species Act. Procedures

for removing the glochidia did not result in sacrificing the

mussel. When glochidia were collected from the mussels,

three subsamples were set aside. The first subsample was

used to test for viability by observing a snapping response

when salt crystals were added to the glochidia (Zale and

Neves 1982). The second subsample of glochidia was used to

determine the longevity of the glochidia once they had been

released. The third subsample of glochidia was preserved in

70% ethanol and later used for scanning electron microscopy.

The remain glochidia were used to preform laboratory

infestations.

The mechanisms for releasing glochidia vary slightly

from species to species, therefore, the techniques used to

collect the glochidia from each mussel species was slightly

different. For example, the gills of Elliptoideus

sloatianus are characterized by reduced swelling that is









17

typically observed in most freshwater mussels with

developing eggs or mature glochidia. As a result, it was

difficult to determine which individuals were gravid under

field conditions, therefore, 10 to 15 individuals were

brought to the laboratory for closer inspection.

Once at the laboratory Elliptoideus sloatianus were

held in plastic tubs on the bottom of large holding tanks

(diameter 3 m; depth 1 m). The mussels were then observed

daily until glochidia were released. When conglutinates

were found on the bottom of the tubs, a subsample was taken

and inspected under the microscope. When fully developed

glochidia were found a viability test was performed as

outlined in Zale and Neves (1982).

Gravid female Lampsilis subangulata were identified by

swollen outer gills with dark pigmentation. When gravid

females were found they were brought back to the laboratory

and placed into a current tank. The current tank contained

substrate, water temperature, and photoperiod similar to the

stream from which the mussels were collected. Lampsilis

subangulata expel glochidia via superconglutinate which made

collecting glochidia easy. Once the females were placed in

the tank, superconglutinates were released within a month.

The search for superconglutinates was conducted in the

stream while collecting female Lampsilis subangulata. The

temperature and number of superconglutinates found were

recorded for each trip.









18

Gravid Medionidus penicillatus were identified in the

field by swollen outer gills. Unlike the other mussels, M.

penicillatus did not abort their glochidia in the laboratory

setting. Glochidia were successfully collected from an

individual mussel by making a small incision in the gill and

washing the glochidia with a squirt bottle into a small

petri dish. Each mussel that underwent this glochidial

removal technique was observed for two to four weeks after

the procedure was performed to determine mussel

survivorship.

Pleurobema pyriforme were the most difficult of the

four mussel species to collect viable glochidia from.

Female P. pyriforme were highly sensitive to handling, and

as a result expelled their glochidia prematurely. In order

to maximize the collection of viable glochidia, female

mussels were not inspected in the field. A collection of 12

to 18 individuals were transported to the laboratory and

placed into three different four-liter holding containers.

The bottoms of the containers were siphoned through a 105 im

mesh sieve and inspected for viable glochidia daily.

Fifteen species of fish representing six families were

used in the host fish identification experiment (Table 2-1).

Fish used in the experiment were purchased from fish

hatcheries or collected in streams located in the

Apalachicola, Chattahoochee, Flint, and Ochlockonee river

drainages. Fish collections were conducted as needed from












Table 2-1. The scientific and common names of the
fish used in the host fish identification experiment.


Scientific Name


CYPRINIDAE
Notropis harper
Notropis petersoni
Notropis texanus
Opsopoeodus emiliae
Pteronotropis hypselopterus


CATOSTOMIDAE
Moxostoma robustum


ICTALURIDAE
Noturus leptacanthus


POECILIIDAE
Gambusia holbrooki
Poecilia reticulata


CENTRARCHIDAE
Lepomis auritus
Lepomis macrochirus
Micropterus punctulatus
Micropterus salmoides


PERCIDAE
Etheostoma edwini
Percina nigrofasciata


Common Name


redeye chub
coastal shiner
weed shiner
pugnose minnow
sailfin shiner



smallfin redhorse



speckled madtom



eastern mosquitofish
guppy



redbreast sunfish
bluegill
spotted bass
largemouth bass



brown darter
blackbanded darter


Scientific nomenclature follows that of Robins
et.al. 1991.









20

April 1995 to July 1996 in Tired Creek, Gad'sden County,

Florida; Jones Creek, Worth County, Georgia; and Bear Creek,

Grady County, Florida (Appendix B-1). Fish were collected

via electroshocking and dip netting from streams with known

low densities of mussels in order to avoid using fish with

prior or existing glochidial attachments (Yeager and Neves

1986). Aerated coolers were used to hold the fish during

transportation to the laboratory. Fish purchased from the

fish farms and collected in the streams were separated by

species and placed into 12 40.0-liter flow through holding

tanks at the USGS laboratory. The fish were held in the

tanks for one to two weeks before the experiments were

conducted allowing the fish to acclimate and any previously

attached glochidia to drop from the fish (Zale and Neves

1982; Luo 1993).

After collecting viable glochidia from a single mussel

species, one to two thousand glochidia were placed into a

500-ml beaker. Larger fish species (Lepomis macrochirus and

Micropterus salmoides) were placed into a 1.0-liter beaker

with 750 ml of water with three to four thousand glochidia.

To avoid overcrowding and reduce stress, no more than six

fish of the same species were placed into the beaker with

the glochidia mixture. The glochidia were suspended by two

to three aeration hoses. Each fish was exposed to the

glochidia mixture for 30 minutes. During this time it was

assumed that each fish had equal opportunity for glochidia

attachment.









21

After 30 minutes the fish were taken from the glochidia

mixture and placed in separate 4.0-liter containers.

Replicates were achieved by holding each fish in a separate

4.0-liter container. Each container held equal volumes of

water, an aeration hose, lid, and a screen on the bottom of

the jar. The screen was used to discourage the fish from

consuming the juvenile mussels after they transformed. The

4.0-liter containers were then placed into a large flow-

through bath to maintain an equal temperature range (20.50 C

to 23.50 C) for all of the fish in the experiment.

Each container was then siphoned through a 6.35-mm tube

and into a 105-um nylon mesh sieve every third day starting

the third day after infestations. When juvenile mussels

were found, the tanks were siphoned daily until no juveniles

were collected for three consecutive days. If no juveniles

were collected one month after the fish were infected with

glochidia, they were inspected under a microscope for

encysted glochidia. If no encysted glochidia were found,

the fish species was deemed a non-host for that mussel

species. If encysted glochidia were found on a live fish,

the fish was returned to the jar and reinspected each week

until no encysted glochidia were found. Fish that died

before the experiment was finished were inspected for

encysted glochidia under a microscope. When encysted

glochidia were found on a dead fish and more than half of

the fish in the experiment died the experiment was run

again.








22

Water temperature, number of sloughed glochidia,

number of transformed glochidia, and dates of glochidia

transformation were recorded for each fish. Transformed

glochidia (juveniles) were defined as free-living glochidia

possessing gill buds, a ciliated foot complete with

protractor and retractor muscles, and a pair of abductor

muscles (Karna and Millemann 1978).

Past experiments (Coker et al. 1921; Zale and Neves

1982; Neves et al. 1991) held fish of the same species in a

single container. This experiment was designed to identify

the number of juvenile mussels produced by each fish

individually. Variations in the number of fish that produce

juvenile mussels and the number of juveniles produced by

each fish can give an indication of the effectiveness of

each fish species in successfully transforming juvenile

mussels for that particular mussel species. A strong host

fish species will have produced juvenile mussels from each

fish used in the experiment.

A primary host fish was determined by the following

criteria: 1) more than 30% of the fish must produce juvenile

mussels, 2) the fish must also have a native distribution

similar to the mussel, 3) and the fish must have similar

microhabitat requirements as the mussel. A fish species

producing juvenile mussel(s) but failing to meet all four

criteria was identified as a satellite host fish. Satellite

host fish are fish that produce juvenile mussels but are not

considered a reliable host fish for the mussel species.









23

Development of Immunity



Development of immunity was tested by infesting the

same fish twice with the same species of glochidia and

recording the number of juveniles produced after each

infestation. The immunity experiment was conducted by

infesting Micropterus salmoides with Lampsilis subangulata

glochidia. M. salmoides were infected with L. subangulata

glochidia because the glochidia were easily obtained and the

glochidia transformed in a short enough time to allow for a

second infestation.

Micropterus salmoides were infested by pipetting

several hundred glochidia into both branchial chambers.

Each M. salmoides was placed into separate containers and

the number of juveniles produced were recorded for each

fish.

Glochidia Reaction to Nonindigenous Fish



Poecilia retuclata were exposed to glochidia from each

of the four target mussel species to test whether

nonindigenous fish have a natural immunity to freshwater

mussels. Poecilia retuclata were infected using the same

methods utilized for the native fishes described in the

primary host identification section.









23

Development of Immunity



Development of immunity was tested by infesting the

same fish twice with the same species of glochidia and

recording the number of juveniles produced after each

infestation. The immunity experiment was conducted by

infesting Micropterus salmoides with Lampsilis subangulata

glochidia. M. salmoides were infected with L. subangulata

glochidia because the glochidia were easily obtained and the

glochidia transformed in a short enough time to allow for a

second infestation.

Micropterus salmoides were infested by pipetting

several hundred glochidia into both branchial chambers.

Each M. salmoides was placed into separate containers and

the number of juveniles produced were recorded for each

fish.

Glochidia Reaction to Nonindigenous Fish



Poecilia retuclata were exposed to glochidia from each

of the four target mussel species to test whether

nonindigenous fish have a natural immunity to freshwater

mussels. Poecilia retuclata were infected using the same

methods utilized for the native fishes described in the

primary host identification section.









24

Glochidia Morphology



Glochidia used for SEM were preserved in 70% ethanol

(Waller et al. 1988b). The preserved specimens were placed

on double-sided carbon adhesive tabs that were positioned

onto mounting pegs. After the specimens were dry (about 15

minutes), three small smudges of graphite cement were placed

at the edge of the carbon tape and the mounting peg. The

mounted glochidia were then coated with 200 angstroms of

gold-pallatium and examined under an ETEC Autoscan scanning

electron microscope. These methods were described by

Hoggarth (1988) and Kennedy et al. (1991). Glochidia shell

surface, lateral side, and micropoints were photographed for

each of the mussel species. Measurements of the glochidia

were made by measuring ten glochidia under a light

microscope.















CHAPTER 3
RESULTS



Reproductive Biology



Mature glochidia were collected from Elliptoideus

sloatianus late February through mid April (1996 and 1997).

This species is a confirmed tachytictic breeder (Table 3-1).

Gravid individuals were collected from the stream with water

temperatures ranging from 8.00 C to 15.0' C. Mature

glochidia were collected from E. sloatianus individuals as

small as 120 mm (anterior edge to posterior edge). Eggs

were released in January 1996 as rigid white conglutinates.

The conglutinates were 10 to 15 mm long and 1.5 mm wide.

Viable glochidia were released in late February 1996 in

the form of white conglutinates that easily disintegrated

when disturbed. Elliptoideus sloatianus glochidia remained

viable for three days after they were released by the female

mussel.

Three potential host fish were identified from nine

species of fish infested with Elliptoideus sloatianus

glochidia: Gambusia holbrooki, Poecilia reticulata, and

Percina nigrofasciata (Table 3-2). Transformation of the

glochidia required 17 to 21 days on Gambusia holbrooki, 16

25















CHAPTER 3
RESULTS



Reproductive Biology



Mature glochidia were collected from Elliptoideus

sloatianus late February through mid April (1996 and 1997).

This species is a confirmed tachytictic breeder (Table 3-1).

Gravid individuals were collected from the stream with water

temperatures ranging from 8.00 C to 15.0' C. Mature

glochidia were collected from E. sloatianus individuals as

small as 120 mm (anterior edge to posterior edge). Eggs

were released in January 1996 as rigid white conglutinates.

The conglutinates were 10 to 15 mm long and 1.5 mm wide.

Viable glochidia were released in late February 1996 in

the form of white conglutinates that easily disintegrated

when disturbed. Elliptoideus sloatianus glochidia remained

viable for three days after they were released by the female

mussel.

Three potential host fish were identified from nine

species of fish infested with Elliptoideus sloatianus

glochidia: Gambusia holbrooki, Poecilia reticulata, and

Percina nigrofasciata (Table 3-2). Transformation of the

glochidia required 17 to 21 days on Gambusia holbrooki, 16

25
































>



4-
0)








-Q
3





























0








Cl)


41







I 0u
0r-





















O
C)


04
4-4




















0)
0)








r-Q


CO
r-







CC


k
ai x x x x


cn cn
r2 1 -3


CC *




> H

1 *H


I H



ao


), U















0
" 4-
i 0V0





S 4-J

















q (D -C
4-1
qC



C0 0


V) 134
4-J








COII -







4J
0





mC) f




4-) -r




II *-
C;
cn 0


















I I I


LnP
u~) ~


O' ->


LO o
CO D 0

LOD, o"
I I I


n in CMN
,_1


I I I


0-0 O
r- 1-


4-4 U)
C
0 (V
i-4




Z -H








ar




S--


,- Ln
Ln m
I I
m LO
-4t2

c O,1


a)
-,D
0














-4 >-







u >

0



U -)
4- C1









0) M

a 1






Uoa
O4-





S4-1 0.
) a)





0 0
3D
a) '
U0


04


U)



a),


( MU
^ -H E



Cu-









28

to 21 days on P. reticulata, and 20 days when encysted on

P. nigrofasciata in an average water temperature of 20.5 C

30 C.

Even though Percina nigrofasciata occur in the same

microhabitat as Elliptoideus sloatianus, P. nigrofasciata

were not considered a primary host fish because only two

glochidia transformed from one P. nigrofasciata (Table 3-3).

A glochidia transformation rate of 16% may not be sufficient

enough to support a wild population of E. sloatianus.

Gravid Lampsilis subangulata were found throughout the

year and are a confirmed bradytictic breeder (Table 3-1).

On 19 May 1995, an individual L. subangulata was discovered

producing a superconglutinate in Cooleewahee Creek, Baker

County, Georgia. This was the first time L. subangulata had

been documented producing a superconglutinate. Lampsilis

subangulata were observed releasing superconglutinates when

water temperatures warmed to about 22.50 C.

Superconglutinates were collected from late May (1995 and

1996) to early July (1995 and 1996) when water temperatures

were between 22.00 C and 23.5 C (Table 3-4). The glochidia

remained viable for about four days after the

superconglutinate was released.

Host fish were identified by infecting 10 species of

fish with Lampsilis subangulata glochidia. Potential host

fish were identified as Gambusia holbrooki, Poecilia

reticulata, Lepomis macrochirus, Micropterus punctulatus,




















O3 T -3 -O
W U '-4 +
C 0
!> ; =I -f m C0
S- O 1 t
-n OOm *
4 0 w m)
o4 U- (1 'I o o)






4-4 -4 ()
u m -a U
r C +1 +1 +1
cr ,,o

EU Z -U 0 0


















S0 ) r -
Co.. co >















00
c;4







co I :
rtl -


























01) ) )0
'4-.dQ C)-.
6 O m 13
4u -0r o ( r


C0 0 co 3 3




























0 0) *0 OS
*C 0

0) 1)
to a













-, I ) -) Q
MCU $ co 00



0 4a-C t7 >
(U 1 *4


r0 4) q 3O

>O *| +jO O






) I ) 0 3























Table 3-4. Dates and temperatures when Lampsilis
subangulata were found releasing superconglutinates
in Cooleewahee Creek, GA.

Number of
Date Superconglutinates Temperature
Found

19 May 1995 4 23.0C
1 June 1995 2 23.0C
22 June 1995* 1 23.0C
23 June 1995* 2 23.0C
16 May 1996 0 20.0C
29 May 1996 5 20.0C
31 May 1996 24 20.0C
6 June 1996 5 23.5C
15 July 1996 1 23.0C
16 July 1996* 1 23.0C
17 July 1996* 1 23.0C

* indicates superconglutinates released in the
laboratory.











and M. salmoides (Table 3-5). Transformation occurred in 13

to 14 days for G. holbrooki, 15 to 17 days for P.

reticulata, 13 days for L. macrochirus, 11 to 15 days for M.

punctulatus, and 12 to 16 days for M. salmoides. The

average water temperature during this transformation period

was 22.50 C 2.5' C.

Only one Lepomis macrochirus out of ten L. macrochirus

tested in the experiment was able to produce one juvenile

Lampsilis subangulata (Table 3-6). The production of a

juvenile mussel from only 10% of the fish is not sufficient

to support the existence of a mussel population. When M.

punctulatus and M. salmoides were exposed to L. subangulata

glochidia 100% of the fish produced juvenile mussel (Table

3-6). Micropterus punctulatus and Micropterus salmoides are

both considered effective host fish for Lampsilis

subangulata. However, the results of a t-test indicated the

M. salmoides was significantly more effective at

transforming L. subangulata glochidia when compared to the

M. punctulatus (df=30; t=-2.49; P=0.019). During the

treatment it was assumed that each fish has equal

opportunity of attaching glochidia and that a point of

saturation was reached.

Gravid Medionidus penicillatus were found at

Chickasawhatchee and Kinchafoonee creeks during the months

of March (1996), April (1996), September (1995), and

November (1996) and is therefore considered a bradytictic















0
O


Ord


CcO
0 m3

4J






Q-4 C
rd 0











(Z








3 3







CD)




C4
r.)










0) H




U)
CO
















COl











.rl


' CD U- C'
P- I Cl ( I


C\' CD r-I -
-i '-I C'\


CO








u co a
000
0 0 *







Ql Q )
CO o C) -1
4- 0 3


0 0C CO0




S So 0
0 O k ^-
04 04 00
ci)C ) *'>-1- *-Ij


mn Ln












0D (Y L 00


C'I C'(i
Q0 m
I I

ZT C'-


mC C'


I I I













1H OM HD
m1 L
~--I CO


Co


u





"u




co
4-






4J 0
4-)
a,


C)


oC 0
O 3

-q c
i )
-14--)1



oa ca
- 4-


a
0 0
C O


I I
I























(4





0
-H

U












0
c-i
7dl)
OH d


0) C

C(1






















o *' 4-)
4 M





















r -l -I
0 c
" H






ci)






DO 0


GR ci
OU
























a4 C)
S-i r-)
C10





















*r-1

Dr C;


0

U r-H +1


0 V m


u
SU



C) -


r-1 -H















ui)-
,- --t


> 0\

-m



DH



00
-J-















r0












H
)H

CO




















0



Cl
H


r-1 -H r\


"4i

0







H 'I



0 5
O 0









34

breeder (Table 3-1). Gravid M. penicillatus were found when

water temperatures were between 11.0' C and 13.0 C.

Medionidus penicillatus were observed at Chickasawhatchee

Creek in March (1996) and April (1996) completely out of the

substrate, laying on their umbos with their white glochidia-

filled gills pushed to the edge of the shell, flapping their

thickened dark mantles (Brim Box, personal communication)

The mantle-flapping behavior could possibly be a mechanism

to attract the proper host fish. This behavior was also

observed on subsequent trips to collect gravid individuals.

Host fish for Medionidus penicillatus were determined by

infesting seven species of fish with thier glochidia.

Glochidia attached to every fish with which they were

exposed. However, Gambusia holbrooki, Poecilia reticulata,

Etheostoma edwini, and Percina nigrofasciata were the only

fish species to produce juvenile mussels (Table 3-7).

Transformation occurred on G. holbrooki in 19 to 21 days, on

P. reticulata in 30 to 32 days, on E. edwini in 30 to 37

days, and on P. nigrofasciata in 29 to 33 days in water held

to an average temperature of 21.50 C. The glochidia

remained viable for only two days after they were released

by the female mussels.

When Etheostoma edwini and Percina nigrofasciata were

exposed to M. penicillatus glochidia, 100% of the E. edwini

and P. nigrofasciata tested transformed M. penicillatus
















0
O
4-}
O

M 0


n4l









0 (1)
Cd









O3
04























4-4 .-
0 )
























)a
303




Cd



N









00r
=3

















-r-1









0U


-r 0l
@lr


mm
I I

M cld


m U


0 ["--

'3 C


c"- .


O c
3 0







0

0
o 0






0 u
E <
ti 0


I I
- cD


Lfn r-


CO
0)


3 *-I

MS c
cm v
on c
SH 0



0 0V




Cj 0
0 C
a J
0 '


O +
*- 0


0

*-1

1 0


{D 0





O

OC
0 I
0 H
0(3
4-) 0D


I I
I I


LO 0

I I
,n c-.-
M r-^









36

glochidia (Table 3-8). Etheostoma edwini and P.

nigrofasciata also occur in the same microhabitat as the

mussel.

Etheostoma edwini and Percina nigrofasciata are both

considered effective host fish for M. penicillatus.

However, the results of a t-test indicate that E. edwini is

significantly more effective at transforming juvenile M.

penicillatus glochidia when compared to P. nigrofasciata

(df=12; t=-6.40; P=0.00003). During thetreatment it was

assumed that each fish had an equal opportunity of attaching

glochidia and that a point of saturation was reached.

Gravid Pleurobema pyriforme were collected at

Chickasawhatchee and Kinchafoonee creeks during the months

of March, April, May, June, and July (1995 and 1996) and are

therefore considered to be tachytictic breeders (Appendix A-

1; Table 3-1). Viable glochidia were collected from P.

pyriforme in water temperatures ranging from 13.0' C in the

spring, to 25.00 C in late summer. The glochidia were found

to remain viable for three days after being released by the

female.

Female Pleurobema pyriforme usually aborted

conglutinates after several days in captivity.

Conglutinates contained glochidia in several stages of

development; eggs, developing eggs, and glochidia. Low

numbers of viable glochidia made the fish infestation

process difficult.


















0 0 C4

- 0 U 0 +u
c- U1-


c; O 4c 0 co a o1
( O CO r CD C in (
*m l UE 0 0 in cu
CL CO U
4-4 w 0 +1 +1 +1 +1

*- 3 -I *- 0 0 c o
i0 ) i n o rn
S -*H (U 0) ) *
,Q > a oo a LO
3 0 -n C






0
11 33







>/0 -I0 0
Z3) 0 -4
' I *-! C 1)







(C ) CL )
OT O 3- C







co t3 00






4-4 -i -W '-I r r1
0O -4 0

( 0 -m g




3 0)4- *H 4
M 4-4 00
-H m





(1) 0 C> C
aO U) 4 M -

01 13
SC



SU) C)0 -H

kt 1 3 ti 0
m ro H 0 3

E -U 0 a 0 0
*- U-J U 4 .
--1 r





00 0 Q 5 --0 +
I D H- rl H 0


co H i H ~ 4 i 43 0
- CO tO c Q O C .r
0 0 D 0 O oc o


r-i co C.-QC 0 U 0
00 c c C H 'D o
-HO 3 *O O U









38

Yokley (1972) also observed the development of glochidia

in multi-stage conglutinates by Pleurobema cordatum. The

production of conglutinates with glochidia in different

developmental stages is thought to be an indication that the

mussel releases a few glochidia at a time instead of the

entire conglutinate (Yokley 1972).

Host fish for Pleurobema pyriforme were identified by

infesting 10 species of fish with P. pyriforme glochidia.

Only the Pteronotropis hypselopterus, Gambusia holbrooki,

and Poecilia reticulata transformed glochidia into juvenile

mussels (Table 3-9). Transformation occurred in 20 to 25

days when attached to the P. hypselopterus, 18 to 21 days

when the glochidia attached to the G. holbrooki, and 19 days

when attached to P. reticulata. The average water

temperature during the transformation period was 23.00 C

3.00 C.

Even though 46% of the Pteronotropis hypselopterus

tested produced juveniles and the number of juveniles

produced by each fish varied, P. hypselopterus are

considered primary host fish for Plerobema pyriforme (Table

3-10).



Development of Immunity



Lampsilis subangulata juveniles were collected from each

Micropterus salmoides after the first and secondinfestations

with glochidia. Only one of the four Micropterus salmoides



















rU


0>1


O








0 H







Ua)









0 (

U-i








)- -4H





























C()
3
a -,





c
S-,--










-,-'-

*r-


QiO Q0


c-


-u
(Q




"r I f) co

"- 0 "'1 O 0
q CO ( Z
) -1 0C O




a)
S T) ( 0



+()J 4U) co ) CO C


LO
CM
I 1 I D

CN













CN











1 I I I
J )o 0n o0
M N LN Um (N


0 (N
i-4


U-~) Cfl
-4c
(Nm


I I
LO (N
Y in


a )

U3 rl
r-l 0


0
0
0C


19
6 0


I I
t I













40





r- D C
rl U +0
4 -+1 DN CD 0
> 0 0 0

O4 4 U Ln Ci Q
3 O O Co rf in N O

+ U4 +1 +1 +1
4- -4 ( .}
o c olo 0 oC






. .c-i ) .
5 O CO O


d -r-l > H (U




) U
JS 0

- I -



coo

HU) -'
:- Q H -HO







31 0

O -

0 V 7-





00
0 C> M ~










*o U KU m a-



H t) y -4 0c i
HQ O
H 4J



IC 0 Q 4
D (l ) H O






4- 3 41









u Z 0 Q>
QI U -1














Scito o 0
'0 (U (
cl 0 4 -U )
















3 -D4 -4 3 C)
aUU)
4- 4- OU 3


44 o C gD









41

produced more juvenile L. subangulata after the second

infestation (Table 3-11). A t-test indicated that there was

a significant reduction in the number of juvenile mussels

after the first infestation when compared to the number of

juveniles produced after the second infestation (df=6;

t=2.01; P=0.04).



Glochidia Reaction to Nonindigenous Fish



Poecilia reticulata successfully transformed glochidia

from Elliptoideus sloatianus, Lampsilis subangulata,

Medionidus penicillatus, and Pleurobema pyriforme. However,

the number of fish producing juveniles and the number of

juveniles produced by each P. reticulata was highly variable

(Table 3-12). None of the four mussel species had glochidia

transform on every P. reticulata infected in the experiment.



Glochidia Morphology



The shape of Elliptoideus sloatianus glochidia is sub-

elliptical (Figure 3-la). The height (dorsal to ventral) is

153 5 pm and the length (anterior to posterior) is 173 6

im. Hooks are lacking and pitting, small holes on the

surface of the shell, is minimal (Figure 3-1b). Shell

sculpturing is loose-looped and increases near the umbo

region. Micropoints, small extensions along the flange,









41

produced more juvenile L. subangulata after the second

infestation (Table 3-11). A t-test indicated that there was

a significant reduction in the number of juvenile mussels

after the first infestation when compared to the number of

juveniles produced after the second infestation (df=6;

t=2.01; P=0.04).



Glochidia Reaction to Nonindigenous Fish



Poecilia reticulata successfully transformed glochidia

from Elliptoideus sloatianus, Lampsilis subangulata,

Medionidus penicillatus, and Pleurobema pyriforme. However,

the number of fish producing juveniles and the number of

juveniles produced by each P. reticulata was highly variable

(Table 3-12). None of the four mussel species had glochidia

transform on every P. reticulata infected in the experiment.



Glochidia Morphology



The shape of Elliptoideus sloatianus glochidia is sub-

elliptical (Figure 3-la). The height (dorsal to ventral) is

153 5 pm and the length (anterior to posterior) is 173 6

im. Hooks are lacking and pitting, small holes on the

surface of the shell, is minimal (Figure 3-1b). Shell

sculpturing is loose-looped and increases near the umbo

region. Micropoints, small extensions along the flange,























Table 3-11. Number of juvenile Lampsilis subangulata
produced from each Micropterus salmoides after two
consecutive infestations.

M. salmoides Number of transformed
size mm juveniles
recovered after

1"' infestation 2nd infestation


45 35 19
51 43 14
58 22 33
64 30 5






















-H
.. -



0







-Iz- *-1.
l 0 C)
U0 0

CO









0 0) 0




-H -I f






-4
Q4 '-1Y
d 1 -
aWQ
300

















CnTi 0-J
* co






















o T) 0
0 CU













S.-i
OH r





















a4 > -Q


C14 -
-40 '-1
-'-1 -1J0

1 41 O













1 0 )
--1 U C










Cn a 'Ol

1 rQ


U *'- tl~








,iwQ 0

cr0 3
E-i C ^


-I O
0
4r -,-rI



0U +1
UO
= 4-4 (















OC
.-4

H CO)



(l)
04

















U) C
0

60




C-)
C l
















3o
M C


SO










m K3 ) ri
N~ ^r N^ 1



















































r- i


Figure 3-1. Glochidium of Elliptoideus sloatianus; a. shell
shape and surface; b. lateral view of valve; c. flange with
micropoints.











cover 15 percent of the flange, a flattened region at the

edge of the shell, region in complete vertical rows (Figure

3-ic).

The glochidia shape of Lampsilis subangulata is sub-

spatulate (Figure 3-2a). The height (dorsal to ventral) is

267 8 pm and the length (anterior to posterior) is 212 7

pm. Hooks are lacking and pitting is present, but few in

number (Figure 3-2b). Shell sculpturing is tight-looped;

and greatest near the umbo region. The micropoints cover

half of the flange region in complete vertical rows (Figure

3-2c).

Medionidus penicillatus glochidia is sub-spatulate

shaped (Figure 3-3a). The height (dorsal to ventral) of the

glochidia is 298 6 pm and the length (anterior to

posterior) is 239 7 pm. Hooks are lacking and pitting is

not apparent (Figure 3-3b). The valve surface is smooth.

Micropoints are blunt lanceolate in shape and decrease in

size from the proximal to distal on the flange (Figure 3-

3c). The micropoints are organized in complete vertical

rows and cover about 75 percent of the flange.

The shape of Pleurobema pyriforme glochidia is sub-

elliptical (Figure 3-4a). The height (dorsal to ventral) of

the glochidia is 172 3 pm and the width (anterior to

posterior) is 164 5 pm. Hooks are lacking and pitting is

apparent on the valve (Figure 3-4b). The surface of the

valve has a slight loose-looped texture near the umbo









46







A














































Figure 3-2. Glochidium of Lampsilis subangulata; a. shell
shape and surface; b. lateral view of valve; c. flange with
micropoints.




























































Figure 3-3. Glochidium of Medionidus penicillatus; a. shell
shape and surface; b. lateral view of valve; c. flange with
micropoints.









48













































Z -._. -











Figure 3-4. Glochidium of Pleurobema pyriforme; a. shell
shape and surface; b. lateral view of valve; c. flange with
micropoints.








49

region. The blunt lanceolate-shaped micropoints are

arranged in an unorganized fashion covering only a quarter

of the flange (Figure 3-4c).















CHAPTER 4
DISCUSSION

Reproductive Biology



A primary host fish for a freshwater mussel is a fish

species that can meet the reproductive requirements and

ensure the existence for a particular mussel species. The

first step of a primary host fish is the need to have

contact with the mussel species. The next requirement for a

primary host fish to meet the reproductive requirements of a

freshwater mussel is the ability to successively transform

glochidia. Freshwater mussels are virtually sessile

organisms. Therefore, the fish species must also frequent

areas with the freshwater mussel in order for the

mussel/fish interaction to occur.

Glochidia from Elliptoideus sloatianus, Lampsilis

subangulata, Medionidus penicillatus, and Pleurobema

pyriforme successfully transformed on Gambusia holbrooki and

Poecilia reticulata. However, G. holbrooki and P.

reticulata are not considered a primary host fish for any of

the target mussel species. Neither G. holbrooki nor the

nonindigenous P. reticulata occupies the same microhabitat

as any of the target mussels species. Gambusia holbrooki















CHAPTER 4
DISCUSSION

Reproductive Biology



A primary host fish for a freshwater mussel is a fish

species that can meet the reproductive requirements and

ensure the existence for a particular mussel species. The

first step of a primary host fish is the need to have

contact with the mussel species. The next requirement for a

primary host fish to meet the reproductive requirements of a

freshwater mussel is the ability to successively transform

glochidia. Freshwater mussels are virtually sessile

organisms. Therefore, the fish species must also frequent

areas with the freshwater mussel in order for the

mussel/fish interaction to occur.

Glochidia from Elliptoideus sloatianus, Lampsilis

subangulata, Medionidus penicillatus, and Pleurobema

pyriforme successfully transformed on Gambusia holbrooki and

Poecilia reticulata. However, G. holbrooki and P.

reticulata are not considered a primary host fish for any of

the target mussel species. Neither G. holbrooki nor the

nonindigenous P. reticulata occupies the same microhabitat

as any of the target mussels species. Gambusia holbrooki









51

and P. reticulata are therefore considered satellite host

fish.

The primary host fish for Elliptoideus sloatianus

remains unknown. Aside from Gambusia holbrooki and Poecilia

reticulata, Percina nigrofasciata was the only other fish

species to transform E. sloatianus glochidia.

The use of only 6 Percina nigrofasciata in the

experiment could have resulted in the low number of

transformed juvenile mussels. Future host fish studies for

Elliptoideus sloatianus should re-examine the potential of

P. nigrofasciata as a primary host fish. The transformation

of two glochidia by P. nigrofasciata may indicate that the

host fish belongs to the Percidae family. Future host fish

experiments should also test other darter species, for

example Etheostoma beani, E. edwini, E. swaini, and E.

parvipinne.

Lampsilis subangulata glochidia transformed on Gambusia

holbrooki, Poecilia reticulata, Lepomis macrochirus,

Micropterus punctulatus, and Micropterus salmoides.

However, only the M. punctulatus and the M. salmoides are

primary host fish. Even though L. macrochirus are found in

the same microhabitat as L. subangulata and were observed

consuming superconglutinates, L. macrochirus are not

considered a primary host fish. Lepomis macrochirus are

considered satellite host fish because they failed to

effectively transform L. subangulata glochidia.









52

The results of the t-test indicated that Micropterus

salmoides are more effective at transforming juvenile L.

subangulata than M. punctulatus. These results may be

misleading because the M. salmoides used in the experiment

were about twice the size as M. punctulatus (Table 3-5).

Lampsilis subangulata glochidia mainly attach to the gill

filaments of Micropterus sp.. Micropterus salmoides have

larger gill chambers compared to M. punctulatus, providing a

larger area for more glochidia to encyst. However, both are

effective at transforming L. subangulata glochidia to

support a population.

The identification of Micropterus punctulatus as a

primary host fish is of special interest because M.

punctulatus is not a native fish to the Apalachicola,

Chattahoochee, Flint, and Ochlockonee river drainages.

Micropterus punctulatus was introduced and is now well

established in the drainage. The ability for M. punctulatus

to act as a primary host fish for Lampsilis subangulata is

reassurance that the introduction of M. punctulatus will not

disrupt the reproductive cycle of L. subangulata.

Medionidus penicillatus glochidia transformed on

Gambusia holbrooki, Poecilia reticulata, Etheostoma edwini,

and Percina nigrofasciata. However, the results of the t-

test indicated that E. edwini are significantly more

effective at transforming M. penicillatus glochidia than P.

nigrofasciata (p=0.0003). These results were unusual since








53

E. edwini used in the experiment were smaller than P.

nigrofasciata (Table 3-7). One reason for these results

could be the glochidia's ability to transform on the gills

and fins of E.edwini. Unlike E. edwini, P. nigrofasciata

appeared to only transform glochidia on its gills. The

transformation of glochidia belonging to the subfamily

Lampsiline on fins is not a common event. However, in some

cases hookless glochidia have been observed encysting on

soft fish fins (Coker et al. 1921).

Under laboratory conditions Etheostoma edwini is the

superior primary host fish but these conclusions may not

hold true in the wild. Other parameters must be considered

before E. edwini can be considered a more effective primary

host fish than P. nigrofasciata. For example, E. edwini may

not be as common as P. nigrofasciata in the wild. If E.

edwini is not common in the field, then it may not be a very

reliable primary host for M. penicillatus.

Etheostoma edwini and Percina nigrofasciata transformed

Medionidus penicillatus glochidia effectively. Even though

the number of juvenile mussels produced varied, 100% of E.

edwini and P. nigrofasciata tested in the experiment

produced juvenile mussels (Table 3-8). The production of

juvenile M. penicillatus by E. edwini and P. nigrofasciata

could support a M. penicillatus population. Under

laboratory conditions E. edwini and P. nigrofasciata are

considered primary host fish for M. penicillatus.









54

Under laboratory conditions Pteronotrcpis hypselopterus

was identified as a primary host fish for Pleurobema

pyriforme. The transformation of glochidia on only 46% of

the Pteronotropis hypselopterus tested in this experiment

may be a common occurrence in the field. Pteronotropis

hypselopterus are common fish that travel in large schools.

A mussel species relying on a fish species that is small in

size and occurs in schools such as P. hypselopterus may not

require a 100% glochidia transformation rate to support the

reproduction of a mussel population. For example, a mussel

species that relies on a host fish species that tends to be

less common and does not occur in schools, like Micropterus

salmoides, must attempt to encyst a large number of

glochidia on as many fish it comes in contact with. These

explanations are only assumptions and require further

studies.



Development of Immunity



The development of immunity was observed when five

Micropterus salmoides were infected with Lampsilis

subangulata glochidia twice. The development of an immune

response has been studied since Coker et al. (1921). Myers

and Millemann (1977) reported a decrease in the ability of

salmonids to produce juvenile Margaritifera margaritifera

after they were infected three times with glochidia. Luo









55

(1993) also reported similar results after darters were

infected three times with glochidia from Medionidus

conradicus. The development of an immunity against

glochidia could be of concern in streams with endangered

mussels and low fish densities. Mussels, such as L.

subangulata that rely on a small number of host fish species

to complete their life cycle, may be at risk of lowered

reproductive capabilities in areas with low numbers of

available host fish. When a species of mussel relies on the

same fish repeatedly for glochidia transformation, the fish

could become more resistant to glochidial attachment. The

length of time that the immune response is effective remains

unknown.



Glochidia Reaction to Nonindigenous Fish



There is a growing concern regarding the introduction of

nonindigenous fish species and how they may affect the

reproductive cycle of North American mussels (Unionids).

The preliminary results of this study indicate that Poecilia

reticulata, a nonindigenous species, could act as a

satellite host fish for the four mussel species tested.

However, P. reticulata is an unreliable satellite host fish.

The number of P. reticulata producing juveniles and the

number of juvenile mussels produced by each P. reticulata

varied for each of the mussel species (Table 3-12). There









56

is speculation that P. reticulata could act as a universal

host fish because this fish species evolved where Unionids

were absent. However, more research is nedded with more

mussel species to confirm these results.

Another nonindigenous fish species that was introduced

into the Apalachicola, Chattahoochee, Flint, and Ochlockonee

river drainages is Micropterus punctulatus. Fortunately, M.

punctulatus was identified as a primary host fish for

Lampsilis subangulata. Micropterus punctulatus appears to

be a comparable primary host fish to M. salmoides, a native

fish in the Apalachicola, Chattahoochee, Flint, and

Ochlockonee river drainages.

These results may indicate that the introduction of

nonindigenous fish into areas where there are native mussels

may not interfere with the freshwater mussel reproductive

cycle. However, more studies must be conducted to confirm

these preliminary results.



Glochidia Morphology



The photos taken of the glochidia from each of the four

mussel species did provide enough information to distinguish

each species. Elliptoideus sloatianus glochidia and

Pleurobema pyriforme glochidia are sub-elliptical in shape

(Figures 3-la and 3-4a, respectively However, E.

sloatianus glochidia have sculpting near the umbo and the









57

shell is more compressed than P. pyriforme glochidia.

Lampsilis subangulata and Medionidus penicillatus glochidia

have similar sub-spatulate shell shapes (Figure 3-2a and 3-

3a). However, one feature that can distinguish the two

species is the highly sculptured umbo region of L.

subangulata glochidia. The umbo region of M. penicillatus

glochidia lacks sculpturing.

Unfortunately, there is not enough information to

distinguish the glochidia of the target mussel species from

any of the other mussel species that occur in the

Apalachicola, Chattahoochee, Flint, and Ochlockonee river

drainages. Elliptoideus sloatianus and Pleurobema pyriforme

have glochidia shapes that might be confused with glochidia

from the genus Elliptio. The glochidia shapes of Lampsilis

subangulata and Medionidus penicillatus are similar to most

of the Lampsiline glochidia in the Apalachicola,

Chattahoochee, Flint, and Ochlockonee river drainages. More

research is needed before the mussel species from the

Apalachicola, Chattahoochee, Flint, and Ochlockonee river

drainages can be identified to the glochidia level.



Conservation Issues



Based on these results, Elliptoideus sloatianus,

Lampsilis subangulata, Medionidus penicillatus, and

Pleurobema pyriforme rely on reproductive mechanisms (lures,









58

i.e., a superconglutinate or mantle flapping) or host fish

that are sensitive to habitat degradation. Lampsilis

subangulata and M. penicillatus rely on lures to visually

attract a host fish. Increased suspended sediments in the

water column reduce visibility thereby rendering host fish-

attracting lures ineffective (Haag et al. 1995). If host

fish are unable to see the lure the interaction between the

mussel and host fish is reduced. As a result, the

reproduction capabilities of these freshwater mussels could

be inhibited.

Mussel species that rely on water currents as a

mechanism to complete their reproductive cycle could be

negatively affected by the construction of a reservoir.

Elliptoideus sloatianus have small, light glochidia that

easily move in the water current. Production of glochidia

easily distributed by water currents could be a mechanism to

carry glochidia to a host fish. Lampsilis subangulata also

relies on water current to effectively display a

superconglutinate to lure the primary host fish.

Reproduction for E. sloatianus and L. subangulata would be

greatly reduced in areas where water currents have been

drastically slowed or damned.

The most important part of the freshwater mussel's

reproductive cycle is the presence of a primary host fish

population. If the host fish population is absent or in low

densities, reproduction for that particular mussel species









59

will be impossible (Weiss and Layzer 1995). Angermeier

(1995) found that fish species belonging to the darter or

minnow groups are more vulnerable to extinction based on

their physiographic range. Mussel species like Medionidus

penicillatus and Pleurobema pyriforme, that rely on fishes

from groups which are more vulnerable to extinction, could

share the increased risk of extinction.









CHAPTER 5
Conclusion



Elliptoideus sloatianus is a tachytictic breeder and is

gravid from late February to April. The primary host fish

for E. sloatianus remains unknown. Satellite host fish were

identified in the laboratory as Gambusia holbrooki and

Poecilia reticulata.

Lampsilis subangulata is a bradytictic breeder and is

gravid throughout the year. Lampsilis subangulata releases

superconglutinates from late May to early July when water

temperatures are between 20.00 C and 23.5 C. The primary

host fish for L. subangulata were identified in the

laboratory as Micropterus punctulatus and Micropterus

salmoides. Satellite host fish were identified in the

laboratory as Gambusia holbrooki, Poecilia reticulata, and

Lepomis macrochirus.

Medionidus penicillatus is a bradytictic breeder and is

gravid September to May. The primary host fish for M.

penicillatus were identified in the laboratory as Etheostoma

edwini and Percina nigrofasciata. Satellite host fish were

identified in the laboratory as Gambusia holbrooki and

Poecilia reticulata.

Pleurobema pyriforme is a tachytictic breeder and is

gravid March to July. The primary host fish was identified

in the laboratory as Pteronotropis hypselopterus. The









61

satellite host fish were identified as Gambusia holbrcoki

and Poecilia reticulata.

Host fish identification for endangered and threatened

mussels is an important part of a recovery plan. By

understanding the specific reproductive requirements of

freshwater mussels (required host fish and mechanisms for

the attachment of glochidia to a host fish), streams with

low densities of host fish or alterations which interfere

with the mussel's ability to attract a host fish can be

identified as streams at risk of losing the mussel

populations. When disturbed streams have been identified

conservation efforts can then be implemented in order to

save the mussel populations as well as the fish populations.

The close association between mussels and fish is just

another example of how ecosystems are interconnected;

conservation efforts should focus on protecting the entire

ecosystem and avoid focusing on a single species.

Many questions surrounding the biology of freshwater

mussels, especially the four proposed mussels in this

experiment, remain unanswered. The host fish for

Elliptoideus sloatianus remains unknown. However, there is

speculation that a fish belonging to the Percidae family may

be a host fish.

The primary host fish for Lampsilis subangulata,

Medionidus penicillatus, and Pleurobema pyriforme need to be

verified in the wild. For example, the host fish identified









62

in this experiment were determined in the laboratory. The

bodies and gills of fish collected, during periods of

glochidia release from streams where the target mussels

occur, must be inspected for encysted glochidia. The

identification of the mussel species to the glochidia level

may be difficult with standard microscopes, but SEM pictures

as used this research, could provide vital information in

identifying the glochidia to the species level. If a

primary host fish were identified in the laboratory and

found to have encysted glochidia belonging to the same

mussel species, that fish would be a confirmed primary host

fish for that mussel species.

The identification of host fish is an important part of

a recovery plan. However, there is also a need to determine

whether existing populations of Elliptoideus sloatianus,

Lampsilis subangulata, Medionidus penicillatus, and

Pleurobema pyriforme are viable. There has been recent

controversy surrounding the sampling techniques for

recruitment of a mussel population. Designing a technique

or method for sampling freshwater mussel recruitment could

be used to test whether conservation efforts or recovery

plans are working for a target mussel species.

















APPENDIX A
SITE RECORDS FOR MUSSEL COLLECTIONS


The following is a list of sites where each mussel species

was collected for this project. The county, state, road

crossing, and drainage is presented for each location site.



















4J 4-J




C U


tfl




CZ3
.H
vU)
-4 -


uE


0r1
O


.-i

Sc

0m


0


u (

U?


cr
r- 4-4
-Hi 0

0
cI)
O
0





0
u U)


O E
* >-




O
U1













C)


*r-H
O 3





4-} H
0


Le





O
U(























APPENDIX B
SITE RECORDS FOR FISH COLLECTIONS


The following is a list of sites where each fish

species was collected for this project. The county, state,

road crossing, and drainage is presented for each location

site.






















T)
4-1


U)
w,



L o


(3 (3


"u






O r
St
0


*H -0

0

44
O
oU






O--
0.,.i


O 4-

o




o.4
0

cU






SD
4r-


(3 Ct


C)


3
-u


'-4 "1
Ct) -l4-


> -c: (1)


al fc


uC E





0


-,

0-4















REFERENCES


Allen, J. D. and A. S. Flecker. 1993. Biodiversity
conservation in running waters. BioScience 43(1)32-43.

Angermeier, P. L. 1995. Ecological attributes of extinction-
prone species: loss of freshwater fishes of Virginia.
Conservation Biology 9: 143-158.

Burch, J. B. 1975. Freshwater Unionacean Clams (Mollusca:
Pelecypoda) North America. Biota of Freshwater
Ecosystems, Identification Manual No. 11, EPA,
Washington D.C.

Butler, R. S. 1989. Distributional records for freshwater
mussels (Bivalvia: Unionidae) in Florida and south
Alabama, with zoogeographic and taxonimic notes.
Walkerana 3(10): 239-261.

Clench, W. J. and R. D. Turner. 1956. Freshwater mollusks of
Alabama, Georgia, and Florida from the Escambia to the
Suwannee River. Bull. Fla. State Museum 1(3):97-237.

Coker, R. E. 1919. Fresh-water mussels and mussel
industries of the United States. Bulletin of the U.S.
Bureau of Fisheries 36: 11-89.

Coker, R. E., A. F. Shira, H. W. Clark, and A. D. Howard.
1921. Natural history and propagation of freshwater
mussels. Bulletin of the U.S. Bureau of Fisheries 37:
75-181.

Couch, C. A., E. H. Hopkins, and P. S. Hardy. 1996.
Influences of environmental settings on aquatic
ecosystems in the Apalachicola-Chattachoochee-Flint
River Basin. U.S. Geological Survey. National Water-
Quality Assessment Program. Water Resources
Investigations Report 95-4278.

Federal Register. 1994. Endangered and threatened wildlife
and plants; Proposed endangered status for five
freshwater mussels and proposed threatened status for
two freshwater mussels from eastern Gulf slope drainage
of Alabama, Florida, and Georgia. Volume 59, No. 148.











Fuller, S. L. H. 1974. Clams and mussels (Mollusca:
Bivalvia), pp. 215-273. In C.W. Hart J. and S. L. H.
Fuller (eds.). Pollution ecology of freshwater
invertebrates. Academic Press, New York.

Haag, W. R., R. S. Butler, and P.D. Hartfield. 1995. An
extraordinary reproductive strategy in freshwater
bivalves: prey mimicry to facilitate larval dispersal.
Freshwater Biology. 34: 471-476.

Heard, W. H. 1975. Determination of the endangered status
of freshwater clams of the Gulf and southeastern states.
Terminal Report for the Office of Endangered Species,
Bureau of Sport Fisheries and Wildlife, U.S. Department
of the Interior. Florida State University, Tallahassee,
Florida. Contract 14-16-000-8905. 31 pp.

Hoggarth, M. A. 1988. The use of glochidia in the
systematics of the Unionidae (Mollusca: Bivalvia). Ph.D.
thesis, Ohio State Univ., Columbia, Ohio. 340 pp.

Howard, A. D. 1951. A river mussel parasitic on a
salamander. Natural History Miscellanea 77: 1-5.

Jansen, W. A. 1990. Seasonal prevalence, intensity of
infestation, and distribution of glochidia of Anodonata
grandis simpsoniana Lea on yellow perch, Perca
flavescens. Canadian Journal of Zoology 69: 964-971.

Jirka, K. J. and Neves, R. J. 1992. Reproductive biology of
four species of freshwater mussels (Mollusca: Unionidae)
in the New River, Virginia and West Virginia. Journal
of Freshwater Ecology 7: 35-44.

Jones, R. 0. 1950. Propagation of fresh-water mussels.
Progressive Fish-Culturist 1: 13-25.

Karna, D. W. and R. E. Millemann. 1978. Glochidiosis of
salmonid fishes. III. Comparative susceptibility to
natural infection with Margaritifera margaritifera (L.)
(Pelycyopda: Margaritanidae) and associated
histopathology. Journal of Parastology 64(3): 528-537.

Kennedy, V. S., S. C. Fuller, and R. A. Lutz. 1991. Shell
Hinge Development of Young Corbicula fluminea (Muller)
(Bivalvia: Corbiculoidea). American Malacological
Bulletin 8: 107-111.

Kondo, T. 1984. Hosts of the larvae of Moncetia lavigeriana
(Bivalvia: Mutelidae) in Lake Tanganyika. Japanese
Journal of Malacology 43:347-352.











Kraemer L. R. 1970. The mantle flap in three species of
Lampsilis (Pelecypoda: Unionidae). Malacologia 10: 225-
282.

Lefevre, G. and W. C. Curtis. 1912. Studies on the
reproduction and artificial propagation of freshwater
mussels. Bulletin of the U.S. Bureau of Fisheries 30:
105-201.

Leitman, H. M., Sohm, J. E. and Franklin, M. A. 1983.
Wetland hydrology and tree distribution of the
Apalachicola River flood plain, Florida. U.S.
Geological Survey water-supply paper 2196-A.

Luo, M. 1993. Host fishes of four species of freshwater
mussels and development of an immune response.
Unpublished Master's Thesis. Tenn. Tech. U. Cookville
19.

Master, L. 1990. The imperiled status of North America
aquatic animals. Biodiversity Network News 3:1-2, 6-7.

Myers, T. R. and R. E. Millemann. 1977. Glochidiosis of
salmonid fishes. I. Comparative susceptibility to
experimental infection with Margaritifera Margaritifera
(L.) (Pelecypoda: Margaritanidae). The Journal of
Parasitology 63:728-733.

Neves, Richard J. 1992. A state of the Unionids. In
Proceedings of The Conservation and Management of
Freshwater Mussels meetings, St. Louis, MO. Oct 12-14.

Ortmann, A. E. 1909. The breeding season of unionidae in
Pennsylvania. The Nautilus 22(10): 99-103.

Robins, R. C., R. M. Bailey, C. E. Bond, J. R. Brooker, E.
A. Lachner, R. N. Lea, and W. B. Scott. 1991. Common and
scientific names of fishes from the United States and
Canada. American Fisheries Society Special Publication
20.

Stansbery, D. H. and C. B. Stein. 1971. Stream
channelization and the preservation of biological
variability: Why naiades (pearly freshwater mussels)
should be preserved. Stream channelization (Part 4).
Hearings before a subcommittee of the Committee on
Government Operations, House of Representatives, Ninety-
second Congress, first session, June 14 :2177-2179.

Tedla, S. and C. G. Fernando. 1969. Observations on the
glochidia of Lampsilis radiata (Gmelin) infesting yellow
perch, Perca flavescens (Mitchell) in the Bay of Quinte,
Lake Ontario. Canadian Journal of Zoology 47:705-712.











Trdan, R. J. and W. R. Hoeh. 1982. Eurytopic host use by two
congeneric species of freshwater mussel (Pelecypoda:
Unionidae: Anodonta). The American Midland Naturalist
108(2) 381-388.

Turgeon, D. D., A. E. Bogan, E. V. Coan, W. K. Emerson, W.
G. Lyons, W. L. Pratt, C. F. E. Roper, A. Scheltema, F.
G. Thompson, and J. D. Williams. 1988. Common and
scientific names of aquatic invetebrates from the United
States and Canada: Mollusks. American Fisheries Society
Special Publication 16.

Waller, D. L. and L. E. Holland-Bartels. 1988. Fish hosts
for glochidia of the endangered freshwater mussel
Lampsilis higginsi Lea (Bivalvia:Unionidae).
Malacological Review 21: 119-122.

Waller, D. L., L. E. Holland-Bartels, and L. G. Mitchell.
1988. Morphology of glochidia of Lampsilis higginsi
(Bivalvia: Unionidae) compared with three related
species. American Malacological Bulletin 6(1): 39-43.

Watters, G. T. 1992. Unionids, fishes, and the species-
area curve. Journal of Biogeography 19: 481-490.

Weiss, J. L. and Layzer J. B. 1995. Infections of glochidia
on fishes in the Barren River, Kentucky. American
Malacological Bulletin 11: 153-159.

Williams, J. D. and R. S. Butler. 1994. Class bivalvia, pp.
53-128. M. Deyup and R. Franz (eds.). In Rare and
endangered biota of Florida. Vol. iv Invetebrates.
University Press of Florida, Gainesville, FL.

Williams, J. D., M. L. Warren, Jr., K. S. Cummings, J. L.
Harris, and R. J. Neves. 1992. Conservation status of
freshwater mussels of the United States and Canada.
Fisheries 18(9): 6-22.

Yeager, B. L. and R. J. Neves. 1986. Reproductive cycle and
fish hosts of the rabbit's foot mussel, Quadrula
cylindrica strigillata (Mollusca:Unionidae) in the upper
Tennessee River drainage. American Midland Naturalist
116(2): 329-340.

Yeager, B. L. and C. F. Saylor. 1989. Fish hosts for
glochidia of Epioblasma brevidens, E. capsaeformis, E.
triquetra and the endangered Quadrula intermedia
(Pelecypoda: Unionidae) from the upper Tennessee River
drainage. Unpublished report. Tennessee Valley
Authority, Norris, Tennessee. 22.









71

Yokley, P. Jr. 1972. Life history of Pleurobema cordatum
(Rafinesque 1820) (Bivalvia: Unionacea). Malacologia 11:
351-364.

Zale, A. V. and R. J. Neves. 1982. Fish hosts for four
species of lampsiline mussels (Mollusca: Unionidae) in
Big Moccasin Creek, Virginia. Canadian Journal of
Zoology 60: 2535-2542.






















BIOGRAPHICAL SKETCH


I was born in Gainesville, Florida on April 15, 1968.

In 1986 I graduated from P.K. Yonge Laboratory School. In

May 1993 I received a B.S.F.R.C degree in natural resource

conservation from the School of Forest Resources and

Conservation at the University of Florida. After completing

an internship with the U.S. Forest Service Fisheries

Department in Washington state, I entered the University of

Florida once again in pursuit of a master's degree in

wildlife ecology and conservation.




University of Florida Home Page
© 2004 - 2010 University of Florida George A. Smathers Libraries.
All rights reserved.

Acceptable Use, Copyright, and Disclaimer Statement
Last updated October 10, 2010 - - mvs