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Differential Allocation in the Flagfish, Jordanella floridae, in Response to Male Condition

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PAGE 1

DIFFERENTIAL ALLOCATION IN THE FLAGFISH, Jordanella floridae IN RESPONSE TO MALE CONDITION By MELISSA ANN NASUTI A THESIS PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLOR IDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE UNIVERSITY OF FLORIDA 2006

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Copyright 2006 by Melissa Ann Nasuti

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iii ACKNOWLEDGMENTS For advice on project design and thesis prep aration I would like to thank Colette St. Mary and Rebecca Kimball. I would also like to show appreciation to Craig Nasuti, Luis Bonachea, Jena, Chojnowski, Hope Klug, and Holly Kindsvater for aiding in the collection of the study species.

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iv TABLE OF CONTENTS page ACKNOWLEDGMENTS.................................................................................................iii LIST OF TABLES...............................................................................................................v LIST OF FIGURES...........................................................................................................vi ABSTRACT......................................................................................................................v ii INTRODUCTION...............................................................................................................1 METHODS........................................................................................................................ ..7 Study Species................................................................................................................7 Study Design.................................................................................................................8 Experimental Manipulation of Male Condition............................................................8 Female Mating Trials....................................................................................................8 Measures of Reproductive Allocation........................................................................10 Color Analysis of Males.............................................................................................11 Lipid Analysis of Males..............................................................................................12 Behavioral Observations of Males..............................................................................12 Statistical Analysis......................................................................................................13 RESULTS........................................................................................................................ ..14 Effects of Diet Manipulation on Ma le Body Size, Condition, Coloration & Behavior.................................................................................................................14 Measures of Reproductive Allocation & Male Food Treatment................................15 Measures of Reproductive Allocation & Male Traits.................................................17 Female Mate Choice...................................................................................................19 Condition-Dependent Expre ssion of Male Traits.......................................................21 DISCUSSION....................................................................................................................4 0 LIST OF REFERENCES...................................................................................................48 BIOGRAPHICAL SKETCH.............................................................................................55

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v LIST OF TABLES Table page 1 Description of nest and non-nest dir ected behaviors recorded for each male flagfish......................................................................................................................2 2 2 The effect of male food trea tment on morphological traits......................................23 3 The effect of male food treatment on measures of c ourtship prior to spawning......24 4 The effect of male food treatment on female reproductive allocation.....................25 5 Results of analyses addressing variation in average egg area and larval length in response to male morphologica l and behavioral traits.............................................26 6 Differrences between mated and unmated males in measures of male body size, fat reserves, condition f actor, and coloration...........................................................27 7 Results of condition-dependent expression analyses...............................................28

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vi LIST OF FIGURES Figure page 1 The relationship between fresh weight and standard length is shown for both high food and low food males..................................................................................29 2 The relationship between dry weight and standard length is shown for both high food and low food males..........................................................................................30 3 The relationship between fat weight a nd standard length is shown for both high food and low food males..........................................................................................31 4 Values of male condition are plotted against residuals from a univariate analysis of co-variance...........................................................................................................32 5 Average larval length (mm), calculated as the average total length of individual larvae within a clutch is plotted against the average hatch date for that clutch.......33 6 Values of male condition are plotted against residuals from a univariate analysis of co-variance...........................................................................................................34 7 Individual egg areas were measured w ithin a clutch and then averaged. These values are plotted against residuals from a univariate analysis of co-variance........35 8 Average proportion of total observation tim e each male spent at their nest( SE) for mated and unmated males prior to receiving eggs.............................................36 9 Average proportion of time spent at the nest fanning ( SE) for mated and unmated males prior to receiving eggs.....................................................................37 10 Frequency of chases ( SE) performe d by mated and unmated males during their recorded observation time prior to receiving eggs...................................................38 11 Relationship between fanning and fat rese rves. Transformed values are show.......39

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vii Abstract of Thesis Presen ted to the Graduate School of the University of Florida in Partial Fulfillment of the Requirements for the Degree of Master of Science DIFFERENTIAL ALLOCATION IN THE FLAGFISH, Jordanella floridae IN RESPONSE TO MALE CONDITION By Melissa Ann Nasuti May 2006 Chair: Colette St. Mary Cochair: Rebecca Kimball Major Department: Zoology The Differential Allocation Hypothesis argu es that females should invest more resources into reproduction when paired with high quality mates as a result of greater direct/and or indirect benefits received fr om reproduction. Evidence suggests that energy reserves often influence male character elabor ation and hence the bene fits received from mate choice. The current study examined the potential for diffe rential allocation by females in response to male condition in the flagfish, Jordanella floridae Male condition was manipulated through diet and was expected to affect measures of male body size, condition (weight/length3), fat reserves, color expressi on, and parental care behavior. Females interacted sequentia lly with two males from th e same or opposite feeding treatment during a two week mating trial. Egg number, egg size, egg energy, hatching success and larval length were recorded as measures of female reproductive allocation. High food males were found to be heavier a nd longer, suggesting th at males prioritize

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viii additional nutrients to growth rather than fa t stores. As expected, high food males showed greater expression of blue and yellow colora tion. Measures of egg number, egg size, egg energy and hatching success were not found to significantly vary with measures of male condition, coloration or behavior. However, males in low condition spent more time fanning, an indicator of parental care. Count er to expectations, male condition had a significant effect on larval leng th as males of a lower weight for a given length fathered larger larvae. This may suggest that females allocate more resources to eggs when mating with males that provide bette r parental care. These result s are consistent with the Differential Allocation Hypothesis suggesting th at female flagfish potentially manipulate reproductive investment in res ponse to mate attractiveness.

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1 INTRODUCTION One of the intriguing issues still facing e volutionary biologists today is to explain the evolution of elaborate sec ondary sexual traits that ofte n appear detrimental to the survival of an individual. Darwin (1871) proposed that such traits do not evolve under natural selection, but via sexua l selection. He argued that the cost of production and maintenance of these traits is offset by an increase in reproductiv e success gained through the acquisition of mates. Typically, males are viewed as the competitive sex and use these traits to compete over access to females or resources for mating. Females are seen as the “choosier sex” and use these traits to discriminate amongst males (Andersson 1994). Considerable evidence from studies of sexua l selection have shown that female mate choice is one of the processes which influences the evolutio n of male traits. Empirical support also suggests that females can furthe r influence trait evol ution by varying the amount of reproductive investment in their offs pring in response to male trait expression (Sheldon 2000). Studies in birds, amphibians, and more recen tly fish, indicate th at females are able to adjust the amount of parental effort (B urley 1988; Szentirmai et al. 2005), manipulate the total number (Petrie & Williams 1993; Reyer et al. 1999; Parker 2003; Edvardsson & Arnqvist 2005), size (Cunningham & Russell 2 000; Kolm 2001; Kolm & Olsson 2003), and sex ratio of eggs produced (Kempenaers et al. 1997; Sheldon et al. 1999), and control egg quality through adjustment of testos terone and immune factors (Gil et al. 1999; Saino et al. 2002a). Adjustments in reprodu ctive investment in response to mate attractiveness

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2 were first tested by Burley in 1986 by manipul ating the perceived attractiveness of a biparental care species of zebra finch, Taeniopygia guttata, with colored leg bands. Burley referred to this concept as the Diffe rential Allocation Hypothesis (DAH). The DAH is based on the premise that the at tractiveness of a mate influences the reproductive value of the breeding attempt with that mate (Sheldon 2000). Differences in male trait expression will lead to selection acting on females to invest more in current reproduction when the investment is balanced by a greater benefit. Attractive high quality mates may affect the payoffs received from reproduction by providing greater direct and/ or indirect benefits to the female and her offspring than unattractive low quality mates. However, greater investment in current reproduction comes at a cost by compromising the amount of available resources for futu re reproduction. Therefore, selection for differential allocation will depend on the magnitude of perceived benefits received from reproduction. Furthermore, the expected future reproductive lifespan of the female and the expected quality of future partners w ill also influence the role of differential allocation. Increased allocation is only expe cted when the current partner is more attractive than expected futu re partners (Sheldon 2000). While attempting to explain the evolution of female mating preferences, models of sexual selection have clearly described th e potential benefits fe males receive through mating. Indicator models propose that male tr aits evolve to “indi cate” or “honestly signal” some aspect of male phenotypic a nd/or genotypic quality (Zahavi 1975; KodricBrown & Brown 1984; Grafen 1990). Male traits become targets of female preference as a result of the correlation between the direct and/or indirect benefits a female receives and the expression of the male trait. Indire ct benefits, commonly referred to as “good

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3 genes” are expected to increase offspring performance through inhe ritance of paternal genes for viability (Norris 1993; Petrie 1994) sexual attractiveness (Gwinner & Schwabl 2005), and parasite resistance (Hamilton & Zuk 1982; Mller 1990). Alternatively, females may directly benefit from non-heritabl e resources that increase female survival or fecundity. Direct benefits include better parental ca re (Petrie 1983; Norris 1990), courtship feeding (Nisbet 1973; Wiggins & Mo rris 1986), nuptial gi fts (Thornhill 1976), or access to resources such as superior nest and oviposition sites (Holm 1973; Campanella & Wolf 1974). For male traits to “honestly” represent the fitness benefits gained by females, theory suggests that male traits must be physiologically costly either to produce or maintain (Andersson 1994). Indicator models of sexual selection, therefore predict the expression of the trait to be condition-depe ndent (Grafen 1990). That is, the degree of trait expression is positively correlated with physical condition. Ma les in better condition signal their quality through great er sexual trait size or more vigorous displays. Males in worse condition incur higher co sts that are associated w ith the production and/or maintenance of these traits and are unable to express the traits to the magnitude that males in better condition can. Evidence for honest advertisement has been found across taxa (Johnstone 1995). In practice, researchers have used measur es of energy reserves (Bolger & Connolly 1989), vigor (Kodric-Brown & Nicoletto 1993), and/or health (Mil inski & Bakker 1990) to estimate an individual’s condition. Em pirical support for the influence of condition measures on the expression of male traits, such as breeding coloration and courtship rates, has been shown in fish. Research of guppi es reveals that several of the traits which

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4 females use to discriminate amongst male s are condition-dependent and function to indicate viability (Nicoletto 1993). For example, male disp lay rate, amount of orange coloration, and ranks of ornamental complex ity were all found to be positively correlated with condition measures (Nicol etto 1993). Breeding coloration has also been shown to be positively correlated with condition measures in sticklebacks (Milinski & Bakker 1990) and in pupfish (Kodric-Brown & Nicoletto 1993). Furthermore, condition measures have been successful in explaining male variation in reproductive success as a result of the in fluence of male energy reserves on parental care. In the sand goby, Pomatoschistus minutus food-supplemented males with higher fat reserves spent more time at their nest car ing for their young. As a result, these males mated sooner and received more eggs than un-supplemented males (Lindstrm 1998). Diet manipulation in small mouth bass also resulted in an increase in reproductive success as a result of food-supplemented males providing longer periods of parental care after swim up of larvae (Ridgway & S huter 1994). Similar patterns of conditiondependent parental care are also seen in the bicolor damselfish, Stegastes partitus as females receive higher quality parental care from more vigorously courting males which is a reflection of male fat reserv es (Knapp & Kovach 1991; Knapp 1995). These experimental results suggest that condition-dependent expression of male secondary sexual traits is common in many fish systems. Furthermore, female preferences for condition-dependent traits ofte n lead to greater direct and/or indirect benefits. According to the DAH, selection s hould favor females who invest more with these mates.

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5 The objective of this study was to determine if female flagfish, differentially allocate reproductive resources in response to male condition. Previous evidence in this system suggests that females may discri minate amongst males based on condition. St. Mary et al. (2001) showed condi tion values of flagfish ma les with low spawning success to decline more drastically over the breed ing season than more successful males. Furthermore, evidence that females are ev aluating males based on traits associated with direct reproductive bene fits makes flagfish an attractive system for the study of differential allocation. Males provide exclus ive parental care of eggs by defending breeding territories from egg predators, oxygenating th eir developing eggs through fanning, and cleaning their nests of debris. Pare ntal care has been shown to be used in mate attraction as a positive relationship wa s found between the numbers of eggs a male received and initial fanning be havior (St. Mary et al. 2001). If the DAH holds, females should allocate resources to more vigorously displaying males who may be in better condition and are able to provi de better parental care. It is likely that parental ca re is a function of male energy reserves in this system. Courtship takes place at the nest with males continually chasing and displaying towards passing females. These activities confine males to their nests and may restrict foraging opportunities to short periods of time inte rspersed between courting females and aggressive interactions with neighboring males. Males ma y therefore be constrained by food availability. This may aff ect their ability to provide pa rental care and subsequently influence female resource allocation. Courtshi p displays and parent al care are known to be energetically costly in other fi sh species (Smith & Wootton 1999).

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6 In this experiment, male condition was e xperimentally manipulated through diet by assigning males to a high or low food treatm ent. This manipulation was expected to affect measures of body size, fat reserves, color expression, and c ourtship/parental care behaviors. Over a two week period, females in teracted sequentially with two males from equal or opposite feeding trea tments. To evaluate differential allocation the total number of eggs spawned with each male was recorded in addition to measures of hatching success, egg energy, egg size, and larval leng th. Under the DAH, females were expected to provide a larger number of eggs and/or hi gher quality eggs to males who have been maintained on high food rations than low food rations.

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7 METHODS Study Species Flagfish, Jordanella floridae are found in fresh, brackis h and salt water systems of central and southern Florida (Foster et al 1969). Flagfish inhabit areas of dense vegetation and live approximately one year (Boschung et al. 2000) Flagfish have a promiscuous mating system. Their breeding se ason extends from late March to early September (Boschung et al. 2000). Males de fend breeding territories from male competitors and egg predators. During courtship, males will repeatedly chase and circle the female. Spawning occurs on submerged vege tation or leaf litter within the breeding territory. During daylight hours, a female may spawn with a single male multiple times or distribute her eggs among males (Mertz & Barlow 1966). Females will lay one egg at a time, releasing approximately 20 or more eggs during a single spawning bout. Males continually accept eggs from multiple female s and provide parental care by fanning, guarding, and cleaning their e ggs (Mertz & Barlow 1966). Ma les continue to care for their eggs until hatching occurs within 45 days of spawning at 25C (Smith 1973). Flagfish show sexual dimorphism in both body size and coloration. Males are larger in size and are the more colorful se x. Males possess horizontal red-orange stripes on their dorsal and anal fins as well as al ong their side. A large dark spot surrounded by yellow is located on their mid side and bl ue iridescence appears on their operculum. During periods of courtship and nest guardi ng, males temporarily express dark coloration

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8 over their entire body. Females are silvery gray in color and are char acterized by a dark spot located at the tail end of their dorsal fin (Page & Burr 1991). Study Design This study was conducted at the University of Florida in Gainesville, Florida. Experimental research was carried out over one breeding season from March to August of 2005. Flagfish for this study were collected during the summer of 2005 from the Otter Creek/Waccasassa River drainage system in northwest central Florida. Sexes were housed separately in 152 liter holding tanks and maintained on a 14L:10D light cycle. The room was kept at a minimum of 27 C. Experimental Manipulation of Male Condition Males were randomly assigned to one of tw o food treatments. In the holding tanks, high food (HF) males were fed a diet of al gae tabs, commercial flake food, and frozen chironomid larvae in excess two times a day. Low food males (LF) received a diet of algae tabs and were fed once every other da y. Males were maintained on the diet for a period of 14 to 42 days prior to interacting wi th a given female. Females were fed in the same manner as high food males in order to maximize their potential for egg production. To control the diet of both sexes during female mating tr ials, males and females were separated by a clear acrylic partition and fed independently at the end of each day. Any remaining food was removed from the tank be fore the start of the mating trial the following morning using a fine mesh net. Female Mating Trials A single female and single male were placed in a 38 liter holding tank. A spawning mat was set in the center of the tank and an artificial plant ( Ludwigia species) was positioned in the back of the tank. The spawning mat was constructed from a square

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9 ceramic tile (10cm x 10cm) with green felt carpe t attached to the top. Male flagfish have been observed in previous research to r eadily accept these spawning mats as breeding territories and typically all e ggs are spawned on the mat. Clear monofilament fishing line was used to separate the spawning mat into 20 square quadrants to aid in the task of counting freshly spawned eggs. Over a ten day period, each female was gi ven the opportunity to interact with two different males for five days each. Each ma le-female pair interacted from 0830 to 1630 hours, for a total of eight hours every day. At the end of each day a perforated clear acrylic partition was placed in the center of the tank between the male and female to prevent spawning overnight. Clear perforated partitions were us ed to facilitate visual and chemical communication between the pair throughout the night. At the end of the first five days the male was removed and replaced with a new male from the opposite or same feeding treatment. Each female was randomly assigned to one of the following four treatments; 1) HF/HF 2) LF/LF 3) HF/LF and 4) LF/ HF. For every ten day period, each treatment was replicated four times. This de sign ensured that the treatments were evenly distributed throughout the breed ing season. Each male and female was used only once. Prior to each five day mating trial, perforat ed clear acrylic partitions were used to physically separate each male-female pair for a two day period. This period of acclimation gave each male a sufficient amount of time to establish his breeding territory over the spawning mat. Prior to interacting in the mating trial, the fish were weighed on an electronic balance to the nearest 0.1g and measured for st andard length to the nearest 0.1cm.

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10 Measures of Reproductive Allocation To assess differential allocation the followi ng five parameters were measured with each male 1) total number of eggs spawned w ith each male 2) average individual egg size per clutch 3) average egg energy content per cl utch 4) hatching success rate of larvae and 5) average larval length per clutch. Spawning mats were checked for eggs f our times a day at approximately 0900 hours, 1100 hours, 1300 hours and 1600 hours. At each time the total number of new eggs was recorded. Freshly spawned eggs were differentiated from older eggs by viewing the developmental stage under a dissecting sc ope and by noting the placement of eggs within the monofilament grid. After the first spawning event, up to 30 eggs were removed from the spawning mat. The remaining eggs were left on the mat to promote continued spawning. If the removal of 30 eggs reduced the total clutch size by more than fifty percent, eggs were removed on subsequent spawning events. Of these eggs, 15 were immediately frozen for energetic analysis. Th e remaining 15 eggs were then placed in a water filled plastic dish and digitally photographed under a dissecting scope. ImageProPlus (Version 4.5.1) software was used to measure the total area (mm2) of each individual egg within a clutc h. To reduce error, three area measurements were recorded and an average of these measurements was used in calculating the average egg area for the entire clutch. The eggs were then pl aced in a 100 ml water filled glass rearing chamber. Each rearing chamber was supplied w ith an air stone and maintained at room temperature of 25C. Hatching success was m easured as the proportion of fertilized eggs hatched by day seven. Once hatched, larvae were digitally photographed by the same methods used for the eggs. Three measurements of total length (mm) were taken for each

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11 larvae and an average of these measurements was used in calculating the average larval length for the entire clutch. To measure the average egg energy cont ent per clutch a dichromate oxidation method was used following the procedure of McEdward and Coulter (1987). Three eggs were randomly chosen from the 15 egg sample. Each egg was incubated for 15 minutes at 110 C in 1ml of 70% phosphoric acid and th en oxidized in 2 ml of 0.30% potassium dichromate for an additional 15 minutes and incubated at the same temperature. Samples were diluted with 3.5ml distilled wate r and energy was measured with a spectrophotometer (440nm) by comparison of th e sample to glucose standards ranging from 0 to 4 joules. An average of the three measurements was calculated to estimate the egg energy content per clutch. Color Analysis of Males To record variation in color expression between males, digital photographs of each male were taken with an Olympus C 2500L camera. To ensure the development of nuptial coloration, photographs we re taken at the end of the first day of each five day mating trial in order to allow males significan t interaction time with females. Each male was hand-netted and placed in a clear water fi lled acrylic box (9cm x 3cm x 6cm) within a standardized photo-box. The small box preven ted the movement of the fish, exposing the lateral side of the male to the camera. Each photo incl uded a red, yellow, and blue color standard. For each photograph we first sa mpled the three color standards recording the hue, saturation, and intensity of each. This allowed for the adjustment of variation in light intensity between images when record ing color measurements. Red was defined as having hue values ranging from 0 to 40 and saturation values ranging from 30 to 100; yellow was defined as hue values from 40 to 55 and saturation values from 70 to 100; and

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12 blue was defined as hue values from 100 to 160 and saturation values from 25 to 100. An image analysis software tool, SigmaScanPr o (Version 5.0.0), was used to measure the mean intensity of red, yellow and blue colo ration for each male. The total area of each color on the fish’s body was also measured a nd a proportion was calcula ted from the area of the entire fish. Lipid Analysis of Males A sample of males from each food treatment were randomly selected and euthenized in MS-222 at the end of the five day female mating trial. Males were frozen and then later dried in an oven at 60 C fo r 24 hours. Upon removal, the dried males were weighed on an electronic balance to the ne arest 0.0001g. To assess differences in male condition as a result of diet manipulation, lipids were extr acted with petroleum ether (Knapp 1995). Males were continually placed in ether until a constant dry weight was achieved. The difference in weight was then ca lculated and the percentage of fat weight/ dry weight was used as a m easure of condition in data an alyses. Fresh weight/length3 was also used as a second condition measure. Behavioral Observations of Males Males were videotaped usi ng a VHS recorder for ten minutes on days one and two of the five day mating trial between 1200 hours and 1600 hours. Flagfish are more active in the afternoon (personal observation). Th is provided an estimate of a male’s reproductive behavior. To evaluate parental ca re, males were also taped for an additional ten minutes in the presence of their first bout of eggs. Male behavior was analyzed using an event recorder program written for this pur pose. Male position was recorded as 1) at the nest or 2) away from the nest. Measured male behaviors include d 1) nest fanning 2) nest cleaning 3) following 4) chasing and 5) spawning (see Table 1 for a more detailed

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13 descriptions of these behavior s). Chasing and cleaning were recorded as counts since the duration of these behaviors is short. Du ration was recorded for fanning, following, and spawning. The percentage of total observation time spent at the nest, away from the nest, following, and spawning was calculated for each male. Frequencies for count behaviors were calculated as the number of times a behavior was performed divided by the total observation time. Fanning and cleaning are behavi ors that occur only while the male is at the nest. Therefore the percentage of time fanning and the frequency of cleaning were calculated from the total time spent at the nest rather than the total ten minute observation time. Statistical Analysis For all analyses, only data from females (a nd their mates) that spawned in at least one week of the two week ma ting trial was considered. Parametric statistical analyses were utilized when the variables met the re quirements of these methods. For all variables normality was tested using the one-sample Kolmogorov-Smirnov Test. If the variables did not meet normality requirements, the appropriate transformations were made. Variables expressed as proportions were arc sin square-root transformed. These included color proportions as well as the proportion of measured fat weight/dry weight (g). The numbers of eggs a female spawned were squa re-root transformed. Male and female fresh weights (g) and standard lengt hs (cm) were log transforme d. For general linear models, variables with p-values greater than 0.15 were removed in a st ep wise fashion until the pvalues that remained were less than this set criterion. Nonparametric statistics were used for behavior data since these variables did not satisfy normality assumptions when transformed. All statistical analyses were performed using SPSS Version 12.

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14 RESULTS Effects of Diet Manipula tion on Male Body Size, Cond ition, Coloration & Behavior Male food treatment was found to have a significant effect on male body size and coloration, though no differences were found for measures of male condition, fat reserves, or behavior. High food males on aver age were significantly heavier and longer than low food males (Table 2). However when controlling for length, condition did not differ. To evaluate the effect of food trea tment on male condition, a multivariate analysis of variance was performed on fresh weight, dry weight, and fat weight with male standard length as a covariate. All weight variables increased with standard length for both food groups as expected (F igures1-3 MANCOVA Fresh Weight F1, 124 = 997.61 P = 0.00; Dry Weight F1, 124 = 473.47. P = 0.00; Fat Weight F1, 124 = 98.78 P = 0.00). Male food treatment had no significant effect (MANCOVA F3, 122 = 1.33 P = 0.27 Wilk’s = 0.97). Furthermore, weight measures did not increase more sharply with body length for high food males than low food males (MANCOVA Food Treatment*Standard Length F3, 122 = 1.36 P = 0.26 Wilk’s = 0.97). Thus, while diet manipul ation did have a significant effect on the weight and standard length of high food males, their body composition remained stable. Comparisons of % fat weight/dry weight showed no significant differences in fat reserves between male food treatments (Table 2). High food males expressed a significantly larger proportion of blue coloration as well as more intense yellow coloration than low food males (Table 2). No differences

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15 were seen between male food treatments for the proportion of red and yellow coloration or the intensities of red a nd blue coloration (Table 2). Although high food males were expected to display more vigorously, no difference was found in the amount of time spent in courtship behavior between male food treatments (Table 3). Furthermore, the pr obability that high food males performed each measured behavior during courtship was not si gnificantly different from that of low food males (Table 3). Behavioral differences we re also expected post-spawning. Male food treatment did not influence the amount of time spent in each behavior once eggs were received (Kruskal-Wallis Test At Nest X2 = 0.78 P = 0.34; Fanning At Nest X2 = 1.41 P = 0.24; Chase Frequency X2 = 0.85 P = 0.34, Following X2 = 0.37 P = 0.54; Cleaning Frequency X2 = 0.11 P = 0.75). Measures of Reproductive Allocation & Male Food Treatment Females were expected to alter their re productive investment in response to male food treatment by allocating a larger number of eggs to high food males. These males were also expected to receive clutches with larger more energetic eggs, a higher hatching success rate and larger larvae. No such patterns were found. To determine female egg allocation pattern s with respect to male food treatment, a repeated measures analysis of variance was us ed to investigate statistical differences in egg numbers between weeks one and two of the mating trial. Two separate analyses were completed, one in which females were paired with males from the same food treatment (LF/LF & HF/HF) and a second in which females were matched with males from separate food treatments (LF/HF & HF/LF). Th e first analysis determined if females on average spawned more eggs when two hi gh food males were received. The second analysis determined if the order of male presentation affected female egg allocation.

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16 Females paired with high food males during both weeks one and two, did not allocate more eggs than females who rece ived only low food males. Female treatment had no significant effect on the average numbe r of eggs spawned over weeks one and two of the mating trial (LF/LF & HF/HF F1, 36 = 1.92 P = 0.18). Furthermore, the difference in eggs spawned between weeks one and two did not vary with female treatment. There was no significant interaction between week and female treatment when considering females matched with males from the same food tr eatment (LF/LF & HF/HF Week*Treatment F1, 36 = 0.00 P = 0.95). The order in which high food ma les were presented had no affect on patterns of egg allocation give n a non-significant interaction between week and treatment when considering females matched with males from separate food treatments (HF/LF & LF/HF Week*Treatment F1, 50 = 0.71 P = 0.40). Females were not limited in their ability to allocate eggs between weeks. That is females that spawn in week one are not more or less likely to sp awn in week two. Egg number did not differ between week one and two of the experiment across female treatments (LF/LF & HF/HF F1, 36 = 0.026 P = 0.872; HF/LF & LF/HF F1, 50 = 0.261 P = 0.612). Given the lack of signifi cant week effects, differences in egg allocation between high food and low food males were directly analyzed in a third repeated measures analysis. In this case, week was ignored, and differences between high food eggs and corresponding low food eggs were compared. Within females there was no significant difference in the number of eggs spawned with a high food versus a low food male ( F1, 50 = 0.15 P = 0.70). Hence, no apparent patterns we re seen in female spawning between male food treatments.

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17 Considering clutches from both weeks one and two, male treatment did not affect hatching success (Table 4). Fu rthermore, when considering only the HF/LF treatment, hatching success did not differ between ma le food treatments (Paired T-test t = -0.30, df = 7 P = 0.78). Low sample size prevented this same analysis with the LF/HF female treatment. Overall, hatching success rate was high for both male food treatments. Across all female treatments, there was a si gnificant difference in egg size between weeks for females that spawned both weeks of the mating trial (Paired T-test t = 2.99 df = 13 P = 0.01). Females spawned larger eggs in week one (1.15mm2 0.03 SE) compared to week two (1.08mm2 0.02 SE) suggesting that female s are limited in producing larger eggs as spawning continues. Given this resu lt, eggs produced during the second week of the mating trial were removed from further an alyses of egg size, egg energy and larval length if the female spawned both weeks. Ma le food treatment did not affect average individual egg size, average egg energy or average larval length (Table 4). Measures of Reproductive Allocation & Male Traits Additional analyses were uti lized to investigate the a ssociation between allocation variables and male morphological and/or behavioral traits. Va riation in egg number, size, and energy among females can not be attribut ed to morphological and/or behavioral aspects of the sire. Larval length shows an exception. The relationship between differences in e gg numbers between week one and two of the experiment with differences in male morphology and behavior between mates was evaluated. A linear relationship was expected, wi th females allocating more eggs to males with greater trait expression. Differences in egg numbers did not co-v ary with differences in male body size, condition, fat rese rves and coloration (ANCOVA Weight F1, 40 = 0.02 P = 0.90; Standard Length F1, 75 = 1.20 P = 0.28; Weight/Length3 F1,74 = 1.16 P = 0.29; %

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18 Fat F1,45 = 0.13 P = 0.72; % Blue F1,43 = 0.08 P = 0.77; Intensity Blue F1,76 = 1.37 P = 0.25; % Red F1,41 = 0.04 P = 0.84; Intensity Red F1,42 = 0.05 P = 0.83; % Yellow F1,73 = 0.02 P = 0.89; Intensity Yellow F1,77 = 2.20 P = 0.14). Nor were any correlations seen with variation in male behavior (At Nest Spearman’s = 0.02 P = 0.85; Fanning At Nest Spearman’s = 0.03 P =0.79; Cleaning Frequency Spearman’s = -0.02 P = 0.90; Chasing Frequency Spearman’s = 0.03 P = 0.80; Following Spearman’s = -0.08 P = 0.53). To determine if females respond to differences in male morphology when allocating eggs of varying size and energy, a un ivariate analysis of covariance was used. Measures of color and male and female condi tion were used as covariates with male treatment as a fixed factor. Male food trea tment did not affect egg size or energy as expected from previous results (ANCOVA Egg Size F1, 40 = 0.18 P = 0.68; Egg energy F1, 34 = 1.75 P = 0.20). Male and female condition a nd blue and red co loration were not found to co-vary with egg size or energy, nor were any significant correlations found between egg size and energy and measures of yellow coloration or aspects of male courtship behavior prior to spawning (Table 5). Male condition and red coloration did not co-vary with egg size however marginal p-va lues were found. Therefore, residuals from the ANCOVA model with proportion red as a covariate were regressed against male condition. The linear regre ssion was non-significant (R2 = 0.05 F1, 60 = 3.25 P = 0.08) however a negative relationship between the variables are seen (Figure 4). Similar analyses to those conducted on egg size and energy were ut ilized for larval length. Measures of average hatch date, color, and male and female condition were used as covariates with male treatment as a fixe d factor. Average hatch date had a significant

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19 positive influence on average larval length (ANCOVA F1,52 = 16.96 P = 0.00; Figure 5 Linear Regression R2 = 0.22 F1,79 = 22.63 P = 0.00). There was no effect of male food treatment (ANCOVA F1 48 = 0.16 P = 0.69) or female condition on average larval length (Table 5). However, male condition showed significant effects on average larval length (ANCOVA F1, 52 = 9.74, P = 0.00.) Residuals from an analysis of variance with average hatch date as a covariate and average larval length as the response were regressed against male condition. A negative relationship was found between average larval length and male condition (Figure 6 Linear Regression R2 = 0.15 F1,65 = 11.64 P = 0.00) similar to that found with egg size analyses These results suggest that males of a lower weight for a given length father larger larvae and received eggs of a greater size. Blue and red coloration were not found to co-vary with larval lengt h nor were any significant correlations found between the residuals from the above analysis and measures of yellow coloration or aspects of male courtship behavior prior to spawning (Table 5). When hatch date is taken into consider ation, egg area has a significant effect on larval length (ANCO VA Average Hatch Date F1,67 = 11.50 P = 0.00; Average Egg Area F1, 67 = 5.06 P = 0.03). Residuals from an analysis of variance of larval length with average hatch date as a covariate were regressed against average egg area. This regression demonstrates a positive relationshi p between average larval length and average egg area (Figure 7 Linear Regression R2 = 0.07 F1, 68 = 4.89 P = 0.03). Therefore, egg size is predictive of larval length. Female Mate Choice High food males were expected to obtain more mates given the prediction that these males would be in better condition. Fe males did not favor mating with one male food treatment over the other. In week one of the mating trial, high food males were

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20 successful in spawning 70% of the time in comparison to low food males with 60% During week one, females were not more likel y to mate with high food males than low food males (Chi-square X2 = 1.09 P > 0.1). The same results are found when considering week two. The probability of mating with hi gh food verses low food males in week two was not significantly different (Chi-square X2 = 0.29, v =1, P > 0.5). High food males had a spawning success rate of 73% while low f ood males showed similar success with 78%. On average, mated males were not signifi cantly heavier or longer than unmated males, nor were they in better condition, ha d greater fat reserves or expressed more coloration (Table 6). Results of behavior analyses do indi cate however, that female flagfish prefer to mate with males who exhi bit signs of parental care during courtship. Prior to receiving eggs, mated males on av erage spent more time defending and fanning their nests (Figures 8-9 Kruskal-Wallis Test At Nest X2 = 7.29 P = 0.01; Fanning At Nest X2 = 5.08 P = 0.02). In addition, mated males chased females more frequently (Figure 10 Kruskal-Wallis Test X2 = 7.19, P = 0.01) than unmated males. Furthermore, the probability that mated males performed these be haviors was greater than that of unmated males (Chi-Square At Nest X2 = 4.18 P < 0.05 ; Fanning At Nest X2 =12.18 P < 0.001; Chase Frequency X2 =7.59 P < 0.5). The proportion of time spent following a female and the frequency of cleaning events did not differ between groups (Kruskal-Wallis Test Following X2 = 0.73 P = 0.39; Cleaning Frequency X2 = 1.93 P = 0.16) nor did the probability with which these behaviors were performed (Chi-Square Following X2 =0.79 P > 0.05; Cleaning Frequency X2 =2.35 P > 0.5). These results are consistent with previous studies which demonstrate that fema le flagfish show preferences for increased parental care during courtship (St. Mary & Li ndstrm in prep; Hale & St. Mary in prep.).

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21 The amount of time males spend fanning duri ng courtship provides females with an a priori expectation of the quality of parental care their offspring will receive. An analysis of covariance, shows a si gnificant influence of egg number ( F1,60 = 23.76 P = 0.000) and time spent fanning pre-spawning ( F1,60 = 9.20 P = 0.00) on fanning at the nest post-spawning. A regression of the residuals from an analysis of covariance with fanning post-spawning as the dependent variable and egg number as a covariate, indicates a significant relationship between fanning pre-spawning and fanning-post spawning (R2 = 0.13 F1,61 = 9.45 P = 0.00), suggesting that mates who fan during courtship are also likely to exhibit the be havior after spawning. Condition-Dependent Expre ssion of Male Traits Diet manipulation was expected to genera te differences in male condition. As a result, males were expected to show condition-dependent expression of potentially costly traits such as courtship be havior and coloration. Of the behaviors measured, fanning the nest and chasing the female possibly require the greatest am ount of energy from the male. Measures of male condition and fat reserves were unable to explain the variation in the frequency of chases a male performed duri ng courtship (Table 7). However, a negative relationship between fanning effort and fat reserves was found when considering only those males who spent time fanning their nests (Figure 11 Linear Regression % Fat Weight F 1, 37 = 3.96 P = 0.05). This relationship was not seen however when considering an additional measure of condition (Linear Regression Weight/Length3 F1, 51 = 0.29 P = 0.60). Measures of male condition and fat rese rves were not predictive of blue and red color expression, nor were they correlated with yellow color expression (Table 7)

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22 Table 1. Description of nest and non-nest di rected behaviors recorded for each male flagfish. Behavior Description Position At the Nest Located within 6cm of vertical distance above the nest with part of the body over the nest. Away From Nest Located away from the nest. Nest – Directed Behavior Fanning Body is angled downward toward nest while performing a rapid side-to-side movement. Cleaning A bite at the nest. Spawning Synchronous movement of fish towards nests. Fish are paired side by side in close proximity, releasing eggs as they make direct contact with the nest. Non – Nest Directed Behavior Following Following of female. Chasing Rapid swimming directed towards female

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23 Table 2.The effect of male food treatment on morphological traits. High Food Average ( SE) Low Food Average ( SE) Test Statistic P Value Measures of Body Size and Condition Fresh Weight (g) 1.97 (0.09) 1.69 (0.08) 5.727 0.02* Standard Length (cm) 3.64 (0.06) 3.46 (0.05) 4.685 0.03* % Fat Weight / Dry Weight 0.19 (0.01) 0.21 (0.01) 0.521 0.47* Measures of Coloration % Body Area Blue 8.56 (0.00) 5.06 (0.00) 6.40 0.01 % Body Area Red 0.04 (0.00) 0.05 (0.00) 0.28 0.60 % Body Area Yellow 0.00 (0.00) 0.00 (0.00) 0.74 0.39 † Intensity Blue 3.54 (0.14) 3.50 (0.16) 0.03 0.86 Intensity Red 2.36 (0.05) 2.29 (0.05) 0.43 0.51 Intensity Yellow 1.33 (0.10) 1.02 (0.11) 5.07 0.02 † *ANOVA (two-tailed) with F Test Statistic † KruskalWallis Test with X2 Test Statistic

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24 Table 3. The effect of male food treatment on measures of courtship prior to spawning. Statistical analyses were used to determine differences between high and low food males for 1) the % of observa tion time a behavior was performed (Kruskal-Wallis Test) 2) the frequency with which a behavior was performed (Kruskal-Wallis Test) and 3) the proba bility that a beha vior was performed (Chi-Square). Measures of Male Behavior Kruskal-Wallis Test Statistic P Value Chi-Square Test Statistic P – Value % Time At Nest 1.85 0.17 1.04 >0.1 Fanning At Nest 0.09 0.76 1.99 >0.1 Following 2.18 0.14 2.04 >0.1 Frequency Cleaning 1.85 0.17 0.06 > 0.5 Chasing 0.34 0.56 0.04 >0.5

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25 Table 4. The effect of male food treatmen t on female reproductive allocation. Measures include hatching success, average egg size, average larval length, and average egg energetic content. Measure of Reproductive Output High Food Average ( SE) Low Food Average ( SE) Test Statistic P Value % Hatched Larvae 86.91 (0.03) 88.90 (0.03) 0.12 0.72† Egg Area (mm2) 1.13 (0.02) 1.12 (0.01) 0.14 0.71* Larval Length (mm) 3.88 (0.06) 3.96 (0.05) 0.96 0.33* Egg Energy (J) 1.88 (0.12) 1.70 (0.12) 1.10 0.30* *ANOVA (two-tailed) with F Test Statistic † KruskalWallis Test with X2 Test Statistic

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26 Table 5. Results of analyses addressing variati on in average egg area a nd larval length in response to male morphologi cal and behavioral traits. Egg Area (mm) Test P Statistic Value Larval Length (mm) Test P Statistic Value Egg Energetics (J) Test P Statistic Value Measures of Condition Male Weight / Length3 3.14 0.08 9.74 0.00* 0.59 0.45 FemaleWeight / Length3 0.28 0.60 0.16 0.69* 2.97 0.09 Measures of Coloration % Body Area Blue 1.79 0.19 3.29 0.08* 0.29 0.59 % Body Area Red 3.28 0.08 0.91 0.34* 0.01 0.91 % Body Area Yellow 0.24 0.10 -0.14 0.31‡ -0.02 0.90‡ Intensity Blue 0.22 0.64 1.52 0.22* 1.50 0.23 Intensity Red 0.10 0.75 1.61 0.21* 0.44 0.51 Intensity Yellow 0.10 0.51 -0.23 0.08‡ -0.09 0.62‡ Measures of Behavior At Nest 0.03 0.81 0.02 0.87‡ -0.01 0.95‡ Fanning At Nest -0.25 0.16 -0.02 0.93‡ 0.02 0.95‡ Cleaning Frequency 0.19 0.30 -0.04 0.84‡ -0.03 0.92‡ ANCOVA with F Test Statistic ‡ Spearman’s Rank Nonparametric Correlation (t wo-tailed) with Correlation Coefficient

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27 Table 6. Differences between mated and unmat ed males in measures of male body size, fat reserves, condition factor, and coloration. Mated Average ( SE) Unmated Average ( SE) Test Statistic P Value Measures of Body Size and Condition Fresh Weight (g) 1.81 (0.08) 1.89 (0.11) 0.77 0.38* Standard Length (cm) 3.54 (0.05) 3.58 (0.07) 0.25 0.62* % Fat Weight / Dry Weight 0.19 (0.00) 0.21 (0.00) 1.87 0.17* Weight / Length3 0.04 (0.00) 0.04 (0.00) 1.78 0.18* Measures of Coloration % Body Area Blue 0.04 (0.02) 0.07 (0.03) 0.00 0.99* % Body Area Red 0.04 (0.01) 0.05 (0.02) 1.31 0.25† % Body Area Yellow 0.00 (0.00) 0.00 (0.00) 1.56 0.21* Intensity Blue 3.52 (0.12) 3.51 (0.20) 0.00 0.96* Intensity Red 2.32 (0.07) 2.34 (0.07) 0.06 0.81† Intensity Yellow 1.24 (0.14) 1.32 (0.14) 0.51 0.47* *ANOVA (two-tailed) with F Test Statistic † KruskalWallis Test with X2 Test Statistic

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28 Table 7. Results of condition-dependent expression analyses. Weight / Length3 Test Statistic P -Value % Fat Weight / Dry Weight Test Statistic P -Value Measures of Coloration % Body Area Blue 0.07 0.79¨ 0.08 0.78¨ % Body Area Red 3.01 0.08¨ 0.06 0.80¨ % Body Area Yellow -0.07 0.38‡ -0.01 0.90‡ Intensity Blue 0.15 0.70¨ 0.08 0.30¨ Intensity Red 0.15 0.70¨ 0.85 0.36¨ Intensity Yellow -0.07 0.37‡ -0.09 0.36‡ Measures of Behavior Fanning At Nest 0.29 0.60¨ 3.96 0.05¨ Chasing Frequency 0.80 0.38¨ 2.38 0.13¨ ‡ Spearman’s Rank Nonparametric Correlation (t wo-tailed) with Correlation Coefficient ¨ Linear Regression wi th F Test Statistic

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29 2.003.004.005.006.00 Standard Length (cm) 0.00 1.00 2.00 3.00 4.00 5.00 6.00Fresh Weight (g) High Food Low Food Figure 1. The relationship between fresh wei ght and standard length is shown for both high food and low food males.

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30 2.003.004.005.006.00 Standard Length (cm) 0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40Dry Weight (g) High Food Low Food Figure 2. The relationship between dry weight and standard length is shown for both high food and low food males.

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31 2.003.004.005.006.00 Standard Length (cm) 0.00 0.10 0.20 0.30 0.40 0.50Fat Weight (g) High Food Low Food Figure 3. The relationship between fat weight an d standard length is shown for both high food and low food males.

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32 Figure 4. Values of male condition are plotted against residuals from a univariate analysis of co-variance in which average egg ar ea was used as the dependent variable and % red coloration of th e sire as a covariate 0.03 0.04 0.04 0.04 0.05 -0.20 -0.10 0.00 0.10 0.20 R 2 = 0.05 Residuals for Average Egg Area (mm2)Male Condition (Fresh Wei ght (g) / Standard Length (cm)) 3

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33 Figure 5. Average larval length (m m), calculated as the average total length of individual larvae within a clutch is plotted agains t the average hatch date for that clutch. Average hatch date was calculated as th e sum of the # of individual larvae times their respective hatch dates di vided by the total number of hatched larvae. 3.00 3.50 4.00 4.50 5.00 5.50 6.00 Average Hatch Date (Days) 3.20 3.40 3.60 3.80 4.00 4.20 4.40 4.60 A verage Larval Length (mm)) R 2 = 0.22

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34 Figure 6. Values of male condition are plotted against residuals from a univariate analysis of co-variance in which average larval length was used as the dependent variable and average hatch date as a covariate. 0.03 0.04 0.04 0.04 0.05 -0.75 -0.50 -0.25 0.00 0.25 0.50 0.75 Residual for Aver age Larval Length (mm) R 2 = 0.15 Male Condition (Fresh Weight (g) / Standard Length (cm) ) 3

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35 Figure 7. Individual egg areas were measured within a clutch and then averaged. These values are plotted against residuals fro m a univariate analysis of covariance in which average larval length was used as the dependent variable and average hatch date as a covariate. 0.90 1.00 1.10 1.20 1.30 1.40 -0.75 -0.50 -0.25 0.00 0.25 0.50 0.75 Residuals for Average Larval Length (mm) R 2 = 0.07 Average Egg Area (mm 2 )

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36 Figure 8. Average proportion of total observati on time each male spent at their nest( SE) for mated and unmated males prior to receiving eggs. Mated Unmated 0.00 0.10 0.20 0.30 % of Observation Time At Nest

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37 Figure 9. Average proportion of time spent at the nest fanning ( SE) for mated and unmated males prior to receiving eggs. Mated Unmated 0.00 0.01 0.02 0.03 0.04 % of Time At Nest Fanning

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38 Figure 10. Frequency of chases ( SE) pe rformed by mated and unmated males during their recorded observation time prior to receiving eggs. Frequency of chases was calculated as the number of chas es performed divided by the total observation time. Mated Unmated 0.00 0.00 0.01 0.01 0.01 Frequency of Chases

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39 Figure 11. Relationship between fanning and fat reserves. Transformed values are show. 0.00 0.10 0.20 0.30 0.40 0.50 0.60 % Fat Weight (g) / Dry Weight (g) 0.10 0.20 0.30 0.40 0.50 0.60 0.70 % of Time At Nest Fanning R 2 = 0.10

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40 DISCUSSION The current study attempted to influe nce male condition through diet. This manipulation was expected to affect measures of body size as well as fat reserves. Male food treatment had a significan t effect on male weight and standard length, as also previously found when a comparable HF/LF diet was implemented (Klug & St. Mary 2005). However, diet manipulation did not in fluence measures of condition. High food males maintained their body composition, as we ight did not increase more sharply with body length. Additional nutrients were utilized for growth as opposed to accumulating in stored fat reserves. This may suggest that body size plays a valuable role in the flagfish mating system. Males may prioritize energy allocation to increased size with the aim of acquiring and defending breeding territories essential fo r reproductive success. In nature flagfish often aggregate and reproductive sites often occur in close pr oximity (R Hale personal observation). Hence, high density conditions are likely. Larger males will be more effective at excluding potential competitor s and thus able to spawn more without disruption, resulting in selection for males to invest in growth. Additionally, female preference for larger dominant males may fu rther enhance these selective pressures. In the current study, variati on in egg number among male s was not related to male size. Nor were mated males found to be of la rger size. However, these results may be influenced by the single male presentation. Fe male preferences in other species have been shown to change in response to their social environment (Kvarnemo et al. 1995; Forsgren

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41 et al. 1996; Kangas & Lindstrom 2001). St. Mary and Lindstrom (unpublished data) suggest a similar response in flagfish as male traits associated w ith competitive ability, such as body size and aggressiveness (freque ncy of chases towards males) became more important in mating and egg allocation decisi ons as the level of male-male competition increased from the absence of competition (s ingle male and single female) to moderate (two males and two females) and high (four males and one female) levels. Males were predicted to show conditiondependent expression of the traits potentially used in mate choice. Measures of color expression were expected to be positively correlated with a male’s physical c ondition. Of the three colors measured, red and yellow were specifically expected to be honest indicators of condition as these have been demonstrated to be carotenoid based in several fish systems (Kodric-Brown 1998). Since carotenoids cannot be synthesized, they must be acquired through the diet (Olson & Owens 1998). As sources are scarce in na ture, the intensity of carotenoid based coloration should reflect the content of a ma le’s diet and his overall foraging ability (Kodric-Brown 1989). In addition, carotenoid s function to support the immune system. Recruitment of carotenoids to non-recoverabl e forms in the dead tissue of scales may compromise a male’s overall health, incr easing his susceptibility to pathogens or parasites (Folstad & Karter 1992). Only male s in superior condition should be able to meet this expense. Carotenoid expression shoul d therefore provide a dependable basis for females to choose males in good condition (Nicoletto 1993). In the current study, measures of male condition were unable to explain the variation in color that existed between males. However, diet manipul ation was successful in influencing aspects of color expression. Diet influe nced the proportion of the body

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42 covered with blue coloration as well as the intensity of yellow coloration, as high food males showed greater expression in each cas e. While, reds, oranges, and yellows are generally thought to represent carotenoid expr ession, pale purples, greens and blues can be produced when carotenoids are bound to proteins (Olson & Owens 1998.) Algae are the primary water-based sources of carote noids for fish, in addition to aquatic invertebrates (Olson & Owens 1998). High f ood males were fed these sources in abundance, suggesting that blue pigments ma y be carotenoid based. Alternatively, blue coloration has been found to be mediated by the expansion an d contraction of melanophores in other fish systems (Kodric -Brown 1996). Measures of red coloration were not affected by diet manipulation, sugge sting that these pigments may originate from other sources. Red, yellow, and orange coloration are also produced by pteridine pigments derived from purines which are synthesized from carbohydr ates and proteins (Hurst 1980). Pteridines have been found to c ontribute to sexual colo ration in poeciliid fishes as orange spots of male guppies were found to contain red pteridine pigments in addition to carotenoids (Grether et al. 2001). In light of these results, it seems that color expression may well be diet dependent. However, it is not a simple reflection of fat reserves for instance. For example, previous research in flagfish has found a significant e ffect of condition (measur ed as the residuals from a regression of log weight to log length) on variance in male color expression in response to temperature and salinity treatmen ts (St. Mary et al. 2001), suggesting that environmental stresses play a role in colo r expression. Furthermore, Johnstone (1995) indicates that color expressi on is typically influenced by both measures of nutritional status and parasite load. Para sites have been found to impe de the uptake of carotenoids

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43 from the gut, limiting the development of co lor expression in fish (Milinski & Bakker 1990; Houde & Torrio 1992). This measure would be of interest to investigate in future research on male color e xpression in flagfish. Measures of parental care and courtship behavior were also expected to show condition-dependent expressi on. Courtship has often been shown to be expensive (Vehrencamp et al. 1989; Hglund et al. 1992) requiring considerable energy reserves to be maintained over time. Thus, a positive re lationship was expected between condition and the frequency and/or time spent performi ng energetically expensive behaviors such as fanning and chasing. No such relationship was found with regard to chasing. However, counter to our expectations fa nning was negatively correlated with fat reserves. This loss in condition could be due to energy expend iture lost through disp laying. Alternatively, this may suggest that males in poor conditi on increased their signaling effort at the expense of future mating as has been dem onstrated in sticklebacks (Candolin 1999). Furthermore, males in low condition may be able to adjust the amount of energy they invest into courtship depending on the social context. For example, Candolin (2000) demonstrated that male sticklebacks increased red coloration in the presence of females and the absence of males when the cost of cheating was low in terms of harassment by dominant males. Male condition may play a more important role in flagfish under different social contexts as male-male competition may ensure honest signaling as also suggested in sand gobies (S vensson & Forsgren 2003). Females were expected to allocate repr oductive resources in response to diet – dependent differences among males. Howeve r, high and low food males were not found to differ significantly in many of the traits measured. Regardless, females were expected

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44 to use variation in morphological characters among males in egg allocation decisions. No such patterns were found. The lack of signifi cant results regarding female egg allocation could be explained by the occurrence of filia l cannibalism as male flagfish are known to consume their eggs (Klug & St. Mary 2005). Ho wever, spawning mats were checked at a minimum of four times a day to prevent fili al cannibalism from influencing egg counts. In fish, egg size has been found to be posit ively associated with larval size and hence offspring survival (Ware 1975; Ma rsh 1986; Pepin 1991; Einum & Fleming 1999). This pattern is also supported from results of the current study where larval length increased with egg area. Thus egg size may be one valuable way in which females benefit from increased investment. Counter to our e xpectations, males of a lower weight for a given length fathered larger larvae. Although, the current st udy found no significant effect of male condition on e gg size, a negative pattern was apparent. These results suggest that females may allocate larger e ggs with more resources to males in poor condition. Granted, male condition had no influence on egg energy, differential investment can not be excluded as resource variation may exist that was not detectable by the general methods used in the current study. For exam ple, the direct transfer of proteins (hormones), immune factors, and mRNA to e ggs and larvae has been demonstrated in oviparous fishes (Heath & Blouw 1998) and in some cases has been correlated with increased larval size and survival (Ays on & Lam 1993). Yolk may be an additional resource to measure directly in future re search as initial yolk volume was found to be correlated with larval length at hatching in capelin, Mallotus villosus (Chambers et al. 1989). Furthermore, Kolm & Olsson (2003) pr opose that female Banggai cardinalfish,

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45 Pterapogon kauderni, re-direct resources into eggs extremely close to spawning, suggesting that females may have more cont rol over egg investment than previously implied. Initially, one may interpret these unexpected results as the female’s adaptive response to low quality males. Females mati ng with unattractive males have been found to increase maternal investment to enhance o ffspring health as a result of a decrease in viability associated with lo w quality fathers (Bluhm & Gowaty 2004). For example, in collared flycatchers, females have been found to compensate for low quality parental care of young sires by increasing yolk testoster one concentrations which boosts begging activity (Michl et al. 2004) and in barn swallows, females manipulated egg carotenoid levels in response to unattractive males whos e offspring may have greater exposure to parasites (Saino et al. 2002b). However in the current study, males with low fat reserves spent more time fanning their nests during courtship, suggesting that fema les allocate larger e ggs and hence larger larvae to those males who are likely to pr ovide better quality pare ntal care and hence possibly greater survivorship of offspring. The proportion of time spent fanning the nest during courtship and when eggs were receiv ed was highly correlated, suggesting that prespawning behavior indicates the quality of parental care a female may expect postspawning. Alternatively, paternal effects can not be ru led out as a source of these unexpected results. Genetic differences among males may have influenced larval length as has been demonstrated in other studies which contribute offspr ing size and growth rates to paternal effect s (Reynolds & Gross 1992).

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46 Consistent with the “good parent process” of sexual selection which argues that parental care functions as a cue in mate choice (Hoelzer 1989), female flagfish also responded to variation in fanning during cour tship in addition to nest defense when selecting mates. The direct benefits associat ed with parental care behaviors have been investigated in the flagfish as nest defens e has been shown to increase egg survivorship (Klug et al. 2005) and fanning is hypothesized to reduce fungal infecti on (St. Mary et al. 2001; 2004) in addition to aiding in the re placement of oxygen. Hence female flagfish select mates that provide better parental care and offer increased offspring survivorship as has been demonstrated in other fish sp ecies (Forsgren 1997; s tlund, & Ahnesj 1998) In summary, the DAH suggests that the at tractiveness of a mate influences the value of the breeding attempt. Females shoul d allocate more resources when mating with an attractive male as the potential for dire ct and/or indirect benefits received from reproduction are greater. Measures of attract iveness such as body size, condition, fat reserves, and color expression did not in fluence female mating and egg allocation decisions. However, male condition had a signif icant affect on larval length, suggesting that females produce eggs that produce larger larvae when mating with males in lower condition. Counter to our expectations, males with low energy reserves fanned more pre and post spawning suggesting that females re spond to variation in parental care by investing more in reproduction. DAH has important implications for the fi eld of sexual selection and should be studied further as it potentially poses a pr oblem for studies that support “good genes” models of sexual select ion. If maternal effect s have not been accounted for, evidence for increased survival of offspring fathered by attractive males may be attributable to

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47 differential female investment rather than paternal genetic effects. Evidence for differential allocation will improve our understa nding of all factors wh ich affect offspring fitness. Furthermore, if conditions offspr ing experience during development influences the expression of sexually sel ected characters in adulthood, differential allocation could have a strong influence on the direction of se xual selected traits within a given species (Qvarnstrom & Price 2001). Despite support for differential allocation in a variety of taxa, experimental tests of the hypothesis in fish is lacking beside s those of Kolm 2001, Kolm & Olsson 2003 and the current study. Given the prev alence of male pare ntal care in fishes (Gross & Sargent 1985) and the importance of perceived benefits in differential allocation theory, future research should aim to expand the number of fish species in which this topic is investigated.

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48 LIST OF REFERENCES Andersson, M. 1994. Sexual Selection Princeton, New Jersey: Princeton University Press. Ayson, F.G., & Lam, T.J. 1993. Thyroxine injections of female rabbitfish ( Siganus guttatus ) broodstock; Changes in thyroid horm onal levels in plasma, eggs, and yolk-sac larvae, and its effect on larval growth and survival. Aquaculture 109, 8393. Bluhm, C.K., & Gowaty, P. 2004. Reproductive compensation for offspring viability deficits by female mallards, Anas platyrhynchos Animal Behaviour 68, 985-992. Burley, N. 1986. Sexual selection for aesthetic traits in species w ith biparental care. American Naturalist 127, 415-445. Burley, N. 1988. The differential-allocati on hypothesis: An experimental test. American Naturalist 132, 611-628. Bolger, T., & Connolly, P.L. 1989. The selectio n of suitable indices for the measurement and analysis of fish condition. Journal of Fish Biology 26, 171-182. Boschung, H.T. Jr., Williams, J.D., Gotshall, D.W., Caldwell, M.C., & Caldwell, D.K. 2000. The Audubon Society Field Guide to North American Fishes, Whales and Dolphins. Chanticleer, New York. Candolin, U. 1999. The relationship between si gnal quality and phys ical condition: is sexual signalling honest in the three-spined stickleback? Animal Behaviour 58, 1261-1267. Candolin, U. 2000. Male-male competition ensu res honest signaling of male parental ability in the three spined stickleback ( Gasterosteus aculeatus ). Behavioral Ecology and Sociobiology 49, 57-61. Campanella, P.J., & Wolf, L.L. 1974. Temporal leks as a mating system in a temperate zone dragonfly (Odonata: Anisoptera). I. Plathemis lydia (Drury). Behaviour 51, 49-87. Chambers, R.C., Leggett, W.C., & Brown, J.A. 1989. Egg size, female effects, and the correlations between early life-history tr aits of capelin, Mallotus villosus : An appraisal at the individual level. Fish Bulletin 87, 515-523.

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49 Cunningham, E.J.A., & Russell, A.F. 2000. Egg investment is influenced by male attractiveness in the mallard. Nature 404, 74-76. Darwin, C. 1871. The Descent of Man and Selection in Relation to Sex Murray, London. Einum, S., & Fleming, I.A. 1999. Maternal effects of egg size in brown trout ( Salmo trutta ): Norms of reaction to environmental quality. Proceedings of the Royal Society of London Series B 266, 2095-2100. Edvardsson, M., & Arnqvist, G. 2005. The eff ects of copulatory courtship on differential allocation in the red flour beetle, Tribolium castaneum Journal of Insect Behavior 18, 313-322. Folstad, I., & Karter, A. J. 1992. Parasites, bright males, and the immunocompetence handicap. American Naturalist 139, 603-622. Forsgren, E. 1997. Female sand gobies prefer good fathers over dominant males. Proceedings of the Royal Society of London Series B 264, 1283-1286. Forsgren, E., Kvarnemo, C., & Lindstrm, K. 1996. Mode of sexual selection determined by resource abundance in two sand goby populations. Evolution 50, 646-654. Foster, N.R., Cairns, J. Jr., & Kaesler, R.L. 1969. The flagfish, Jordanella floridae, as a laboratory animal for beha vioral bioassay studies. Proceedings of the Academy of Natural Sciences Philadelphia, 121, 129-152. Gil, D., Graves, J., Hazon, N., & Wells, A. 1999. Male attractiveness and differential testosterone investment in zebra finch eggs. Science 286, 126-128. Grafen, A. 1990. Biological signals as handicaps. Journal of Theoretical Biology 144, 517-546. Grether, G.F., Hudon, J., & Endler, J.A. 2001. Carotenoid scarcity, synthetic pteridine pigments and the evolution of sexual coloration in guppies ( Poecilia reticulata ). Proceedings of the Royal Society of London, Series B 268, 1245-1253. Gross, M.R., & Sargent, R.C. 1985. The evolut ion of male and fema le parental care in fishes. American Zoologist 25, 807-822. Gwinner, H. & Schwabl, H. 2005. Evidence for sexy sons in European starlings ( Sturnus vulgaris ). Behavioral Ecology and Sociobiology 58, 375-382. Hamilton,W.D., & Zuk, M. 1982. Heritable true fitness and bright birds: A role for parasites? Science 218, 384-386. Heath, D.D., & Blouw, D.M. 1998. Are maternal effects in fish adaptive or merely physiological side effects? In Maternal Effects as Adaptations (Mousseau, T.A., & Fox, C.W. eds) pp 178-201. New York: Oxford University Press.

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50 Hoelzer, G.A. 1989. The good parent process of sexual selection. Animal Behaviour 38, 1067-1078. Hglund, J., Kls, J.A., & Fiske, P. 1992. The costs of secondary sexual characters in the lekking great snipe. ( Gallinago media ). Behavioral Ecology and Sociobiology 30, 309-315. Holm, C.H. 1973. Breeding sex ratios, territoria lity, and reproductive success in the redwinged blackbird ( Agelaius phoeniceus ). Ecology 54, 356-365. Houde, A.E., & Torio, A.J. 1992. Effect of para sitic infection on male colour pattern and female choice in guppies. Behavioral Ecology 3, 346-351. Hurst, D.T. 1980. An Introduction to the Chemistry and Biochemistry of Pyrimidines, Purines and Pteridines New York: John Wiley. Johnstone, R.A. 1995. Sexual selection: Honest advertisement and the handicap principle: Reviewing the evidence. Biological Reviews 70, 165. Kangas, N., & Lindstrm, K. 2001. Male interac tions and female mate choice in the sand goby, Pomatoschistus minutus Animal Behaviour 61, 425-430. Kempenaers, B., Verheyen, G.R., & Dhondt, A. A. 1997. Extra-pair paternity in the blue tit ( Parus caeruleus ): Female choice, male charact eristics, and offspring quality. Behavioral Ecology 8, 481-492. Klug, H., & St.Mary, C.M. 2005. Reproductive fitn ess consequences of filial cannibalism in the flagfish, Jordanella floridae Animal Behaviour 70, 685-691. Klug, H., Chin, A., & St. Mary, C.M. 2005. The net effects of guarding on egg survivorship in the flagfish, Jordanella floridae Animal Behaviour 69, 661-668. Knapp, R.A. 1995. Influence of energy reserves on the expression of a secondary sexual trait in male bicolor damselfish, Stegastes partitus Bulletin of Marine Science 57, 672-681. Knapp, R.A., & Kovach, J.T. 1991. Courtship as an honest indicator of male parental quality in the bicolor damselfish, Stegastes partitus. Behavioral Ecology 2, 295300. Kodric-Brown, A. 1989. Dietary carotenoids and male mating success in the guppy; and environmental component to female choice. Behavioral Ecology and Sociobiology 25, 393-401. Kodric-Brown, A. 1996. Role of male-male competition and female choice in the development of breeding coloration in pupfish ( Cyprinodon pecosensis ). Behavioral Ecology 7, 431-437.

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51 Kodric-Brown, A. 1998. Sexual dichromatis m and temporary color changes in the reproduction of fishes. American Zoology 38, 70-81. Kodric-Brown, A., & Brown, J.J. 1984. Truth in advertising: The kinds of traits favored by sexual selection. American Naturalist 124, 309-323. Kodric-Brown, A. & Nicoletto, P.F. 1993. Th e relationship between physical condition and social status in pupfish, Cyprinodon pecosensis Animal Behaviour 46, 12341236. Kolm, N. 2001. Females produce larger eggs for large males in a paternal mouth brooding fish. Proceedings of the Royal Society of London Series B 268, 22292234. Kolm, N., & Olsson, J. 2003. Differential inve stment in the Banggai cardinalfish: Can females adjust egg size close to egg matura tion to match the attractiveness of a new partner? Journal of Fish Biology 63, 144-151. Kvarnemo, C., Forsgren, E., & Magnhagen, C. 1995. Effects of sex ratio on intraand inter sexual behavior in sand gobies. Animal Behaviour 50, 1455-1461. Lindstrm, K. 1998. Energetic constraint s on mating performance in the sand goby. Behavioral Ecology 9, 297-300. Marsh, E. 1986. Effects of egg size on offspr ing fitness and maternal fecundity in the orange-throat darter, Etheostoma spectabile (Pisces: Percidae). Copeia, 18-30. Mertz, J.C., & Barlow, G.W. 1966. On the reproductive behavior of Jordanella floridae (Pisces: Cyprinodontidae) with special reference to a quantitative analysis of parental fanning. Zeitschrift fr Tierpsychologie 23, 537-554. McEdward, L.R., & Coulter, L.K. 1987. E gg volume and energetic content are not correlated among sibling offspring of starfi sh: Implications for life-history theory. Evolution 41, 914-917. Michl, G., Trk, J., Pczely, P., Garamszegi, L.Z., & Schwabl, H. 2004. Female collared flycatchers adjust yolk testosterone to male age, but not to attractiveness. Behavioral Ecology 16, 383-388. Milinski, M., & Bakker, C.M. 1990. Female sticklebacks use male coloration in mate choice and hence avoid parasitized males. Nature 344, 330-333. Mller, A.P. 1990. Effects of a haemat ophagous mite on the barn swallow ( Hirundo Rustica ): A test of the Hamilton and Zuk hypothesis. Evolution 44, 771-784. Nicoletto, P.F. 1993. Female sexual response to condition-dependent ornaments in the guppy, Poecilia reticulata Animal Behaviour 46, 441-450.

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52 Nisbet, I.C.T. 1973. Courtship-feeding, egg-si ze, and breeding success in common terns. Nature 241, 141-142. Norris, K. 1990. Female choice and the quali ty of parental care in the great tit, Parus major Behavioral Ecology and Sociobiology 27, 275-281. Norris, K. 1993. Heritable variation in a plumag e indicator of viability in male great tits, Parus major Nature 362, 537-539. Olson, V.A., & Owens, I.P.F. 1998. Costly sexual signals: Are carote noids rare, risky or required? Trends in Evolution and Ecology 13, 510-514. stlund, S., & Ahnesj, I. 1998. Female fifteen-spi ned sticklebacks prefer better fathers. Animal Behaviour 56, 1177-1183. Page, L. M., & Burr, B.M. 1991. A Field Guide to Freshwater Fishes: North America North of Mexico. New York: Houghton Mifflin Company. Parker, T.H. 2003. Genetic benefits of mate choice separated from differential maternal investment in red junglefowl ( Gallus gallus ). Evolution 57, 2157-2165. Petrie, M. 1983. Female moorhens compete for small fat males. Science, 220, 413415. Petrie, M. 1994. Improved growth and surviv al of offspring of peacocks with more elaborate trains. Nature 371, 598-599. Petrie, M., & Williams, A. 1993. Peahens lay more eggs for peacocks with larger trains. Proceedings of the Royal Society of London Series B 251, 127-131. Pepin, P. 1991. Effects of temperature and size on development, mortality, and survival rates of the pelagic early lif e-history of marine fish. Canadian Journal of Fisheries and Aquatic Science 48, 503-518. Qvarnstrm, A., & Price, T.D. 2001. Maternal effects, paternal effects and sexual selection. Trends in Ecology and Evolution 16, 95-100. Reyer, H.U., Frei, G., & Som, C. 1999. Cryptic female choice: frogs reduce clutch size when amplexed by undesired males. Proceedings of the Royal Society of London Series B 266, 2101-2107. Reynolds, J.D., & Gross, M.R. 1992. Female ma te preference enhances offspring growth and reproduction in a fish, Poecilia reticulata Proceedings of the Royal Society of London Series B 250, 57-62. Ridgway, M.S., & Shuter, B.J. 1994. The eff ects of supplemental food on reproduction in parental male smallmouth bass. Environmental Biology of Fishes 39, 201-207.

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53 Saino, N., Paola Ferrari, R., Martinelli, R ., Romano, M., Rubolini, D., & Mller, A.P. 2002a. Early maternal effects mediat ed by immunity depend on sexual ornamentation of the male partner. Proceedings of the Royal Society of London Series B 269, 1005-1009. Saino, N., Bertacche, V., Paola Ferrari, R., Ma rtinelli, R., Pape Moller, A., & Stradi, R. 2002b. Carotenoid concentration in barn sw allow eggs is influenced by laying order, maternal infection and paternal ornamentation. Proceedings of the Royal Society of London Series B 269, 1729-1733. Sheldon, B.C., 2000. Differential allocation; Tests, mechanisms, and implications. Trends in Ecology and Evolution 15, 397-401. Sheldon, B.C., Andersson, S., Griffith, S.C., rnborg, J., & Sendecka, J. 1999. Ultraviolet colour variation in fluences blue tit sex ratios. Nature 874-877. Smith, W.E. 1973. A Cypriodontid fish, Jordanella floridae as a laboratory animal for rapid chronic bioassays. Journal of the Fisheries Research Board of Canada, 30, 329-330. Smith, C., & Wootton, R. J. 1999. Parental ener gy expenditure of th e male three-spined stickleback. Journal of Fish Biology 54, 1132-1136. St. Mary, C.M., Noureddine, C.G., & Lindstr m, K. 2001. Environmental effects on male reproductive success and parental care in the Florida flagfish Jordanella floridae Ethology 107, 1035-1052. St. Mary, C.M., Gordon, E., & Hale, R. E. 2004. Environmental effects on egg development and hatching success in Jordanella floridae a species with parental care. Journal of Fish Biology 65, 1-9. Svensson, O., & Forsgren, E. 2003. Male mating success in relation to food availability in the common goby. Journal of Fish Biology 62, 1217-1221. Szentirmai, I., Komdeur, J., & Szkely, T. 2005. What makes a nest-building male successful? Male behavior and fe male care in penduline tits. Behavioral Ecology 994-1000. Thornhill, R. 1976. Sexual selection and nuptial feeding behaviour in Bittacus apicalis (Insecta, Mecoptera). American Naturalist 110, 529-548. Vehrencamp, S.L., Bradbury, J.W., & Gibson, R.M. 1989. The energetic cost of display in male sage grouse. Animal Behaviour 38, 885-896. Ware, D.M. 1975. Relation between egg size, grow th, and natural mortality of larval fish. Journal of the Fisheries Research Board of Canada 32, 33-41.

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54 Wiggins, D.A., & Morris, R.D. 1986. Criteria for female choice of mates: Courtship feeding and parental ca re in the common tern. American Naturalist, 128, 126-129. Zahavi, A. 1975. Mate selectiona selection for a handicap. Journal of Theoretical Biology 53, 205-214.

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55 BIOGRAPHICAL SKETCH In June of 2003 I received a Bachelor of Science degree from Ohio State University. As an undergraduate I was able to focus on three major research projects in various disciplines of zoology. These projec ts occurred in the areas of neuroethology, pharmacology, aquatic ecology, and mating systems. I began at the Rothenbuhler Honey Bee Lab, assisting in research aimed at understanding the role of nitric oxide in the feeding behavi or of the adult sphinx moth, Manduca sexta In April of 2002, I presented this research at the 24th Annual Meeting for the Association of Chemoreception Sciences in Sarasota, Florida. In May of 2002, I continued to present this re search at several undergraduate research symposiums. As a result, I received the Arts and Sciences Award for Excellence in Scholarship and was given the opportunity to travel abroad in November of 2002 to attend the University of Sao Paolo’s Symposium of Underg raduate Research in Brazil. Following this experience, I worked as a gr aduate student assist ant at the Aquatic Ecology Lab. I gained invaluable field expe rience working in Ohio’s reservoirs, aiding in research that sought to determine the im pact of stocked Saugeye on the abundance of their primary prey, the Gizzard Shad. I became familiar with techniques used to collect larval and adult fish, in addition to methods used for ageing fish. Throughout my undergraduate years I voluntee red in the lab of Jerry Downhower. This research focused on the costs of comp romised female choice and the consequences of estrogen-like compounds on mating behavior in Japanese killifish, Oryzias latipes It

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56 was from this experience that I became in terested in the study of sexual selection. I decided to pursue my interests further by atte nding graduate school at the University of Florida and studying the mati ng behavior of flagfish, Jordanella floridae in the lab of Colette St. Mary. As an undergraduate I was grateful to be given the opportunity to participate as a research assistant in several laboratories. As a graduate student with my own research program I have extended this opportunity to students at the University of Florida by mentoring two undergraduates. Each student aide d in the collection of data. Furthermore, over the past eight semesters I have also interacted with the student body by teaching laboratory courses in introductory biology, e volution and ecology. I now hope to pursue a career in science education.


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Title: Differential Allocation in the Flagfish, Jordanella floridae, in Response to Male Condition
Physical Description: Mixed Material
Copyright Date: 2008

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Source Institution: University of Florida
Holding Location: University of Florida
Rights Management: All rights reserved by the source institution and holding location.
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DIFFERENTIAL ALLOCATION IN THE FLAGFISH, Jordanellafloridae, IN
RESPONSE TO MALE CONDITION
















By

MELISSA ANN NASUTI


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


2006

































Copyright 2006

by

Melissa Ann Nasuti















ACKNOWLEDGMENTS

For advice on project design and thesis preparation I would like to thank Colette St.

Mary and Rebecca Kimball. I would also like to show appreciation to Craig Nasuti, Luis

Bonachea, Jena, Chojnowski, Hope Klug, and Holly Kindsvater for aiding in the

collection of the study species.
















TABLE OF CONTENTS

page

A C K N O W L E D G M E N T S ......... .................................................................................... iii

L IST O F T A B L E S .............................................................................................. v

LIST O F FIG U R E S .... .............................. ....................... ........ .. ............... vi

ABSTRACT ........ .............. ............. .. ...... .......... .......... vii

INTRODUCTION ............................... .................... .............

M E T H O D S ............................................................................ . 7

Stu dy Sp ecies ................................................ 7
Study Design...................................................... ..... ............... ...............
Experimental Manipulation of Male Condition........................................................8
Fem ale M ating Trials ............................................................ ... ... ....8
M measures of Reproductive Allocation .................................. ............ .................. 10
Color A analysis of M ales ......................................... ...... .. .... .. .............. .. 11
Lipid A analysis of M ales .................. ........................... .................... 12
B ehavioral Observations of M ales..................................... .......................... ......... 12
Statistical A nalysis................................................... 13

R E S U L T S ................................................................................14

Effects of Diet Manipulation on Male Body Size, Condition, Coloration &
B behavior .................. .......... .. ......... ..... .... ..............................14
Measures of Reproductive Allocation & Male Food Treatment .............................15
Measures of Reproductive Allocation & Male Traits..............................................17
Female M ate Choice ............................. ....................... ... ............... 19
Condition-Dependent Expression of M ale Traits...................................................21

D ISC U SSIO N ............... .................................... ...........................40

LIST OF REFEREN CE S ........................................ ........................... ............... 48

B IO G R A PH IC A L SK E TCH ..................................................................... ..................55

















LIST OF TABLES


Table p

1 Description of nest and non-nest directed behaviors recorded for each male
fla g fish ............................... ......... ...... ..................... ................ 2 2

2 The effect of male food treatment on morphological traits.................................23

3 The effect of male food treatment on measures of courtship prior to spawning......24

4 The effect of male food treatment on female reproductive allocation.. ..................25

5 Results of analyses addressing variation in average egg area and larval length in
response to male morphological and behavioral traits.........................................26

6 Differrences between mated and unmated males in measures of male body size,
fat reserves, condition factor, and coloration ................................. ............... 27

7 Results of condition-dependent expression analyses. ............................................28















LIST OF FIGURES


Figure page

1 The relationship between fresh weight and standard length is shown for both
high food and low food m ales. ........................................................................... 29

2 The relationship between dry weight and standard length is shown for both high
food and low food m ales. ........................................ ................................. 30

3 The relationship between fat weight and standard length is shown for both high
food and low food m ales. ...... ........................... ........................................31

4 Values of male condition are plotted against residuals from a univariate analysis
of co-variance. ....................................................................... 32

5 Average larval length (mm), calculated as the average total length of individual
larvae within a clutch is plotted against the average hatch date for that clutch.......33

6 Values of male condition are plotted against residuals from a univariate analysis
of co-variance. ....................................................................... 34

7 Individual egg areas were measured within a clutch and then averaged. These
values are plotted against residuals from a univariate analysis of co-variance........ 35

8 Average proportion of total observation time each male spent at their nest( + SE)
for mated and unmated males prior to receiving eggs. .........................................36

9 Average proportion of time spent at the nest fanning ( SE) for mated and
unmated males prior to receiving eggs .... ........... ........................................ 37

10 Frequency of chases ( SE) performed by mated and unmated males during their
recorded observation time prior to receiving eggs.. ........................................... 38

11 Relationship between fanning and fat reserves. Transformed values are show.......39















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

DIFFERENTIAL ALLOCATION IN THE FLAGFISH, Jordanellafloridae, IN
RESPONSE TO MALE CONDITION


By

Melissa Ann Nasuti

May 2006

Chair: Colette St. Mary
Cochair: Rebecca Kimball
Major Department: Zoology

The Differential Allocation Hypothesis argues that females should invest more

resources into reproduction when paired with high quality mates as a result of greater

direct/and or indirect benefits received from reproduction. Evidence suggests that energy

reserves often influence male character elaboration and hence the benefits received from

mate choice. The current study examined the potential for differential allocation by

females in response to male condition in the flagfish, Jordanellafloridae. Male condition

was manipulated through diet and was expected to affect measures of male body size,

condition (weight/length3), fat reserves, color expression, and parental care behavior.

Females interacted sequentially with two males from the same or opposite feeding

treatment during a two week mating trial. Egg number, egg size, egg energy, hatching

success and larval length were recorded as measures of female reproductive allocation.

High food males were found to be heavier and longer, suggesting that males prioritize









additional nutrients to growth rather than fat stores. As expected, high food males showed

greater expression of blue and yellow coloration. Measures of egg number, egg size, egg

energy and hatching success were not found to significantly vary with measures of male

condition, coloration or behavior. However, males in low condition spent more time

fanning, an indicator of parental care. Counter to expectations, male condition had a

significant effect on larval length as males of a lower weight for a given length fathered

larger larvae. This may suggest that females allocate more resources to eggs when mating

with males that provide better parental care. These results are consistent with the

Differential Allocation Hypothesis suggesting that female flagfish potentially manipulate

reproductive investment in response to mate attractiveness.















INTRODUCTION

One of the intriguing issues still facing evolutionary biologists today is to explain

the evolution of elaborate secondary sexual traits that often appear detrimental to the

survival of an individual. Darwin (1871) proposed that such traits do not evolve under

natural selection, but via sexual selection. He argued that the cost of production and

maintenance of these traits is offset by an increase in reproductive success gained through

the acquisition of mates. Typically, males are viewed as the competitive sex and use these

traits to compete over access to females or resources for mating. Females are seen as the

"choosier sex" and use these traits to discriminate amongst males (Andersson 1994).

Considerable evidence from studies of sexual selection have shown that female mate

choice is one of the processes which influences the evolution of male traits. Empirical

support also suggests that females can further influence trait evolution by varying the

amount of reproductive investment in their offspring in response to male trait expression

(Sheldon 2000).

Studies in birds, amphibians, and more recently fish, indicate that females are able

to adjust the amount of parental effort (Burley 1988; Szentirmai et al. 2005), manipulate

the total number (Petrie & Williams 1993; Reyer et al. 1999; Parker 2003; Edvardsson &

Arnqvist 2005), size (Cunningham & Russell 2000; Kolm 2001; Kolm & Olsson 2003),

and sex ratio of eggs produced (Kempenaers et al. 1997; Sheldon et al. 1999), and control

egg quality through adjustment of testosterone and immune factors (Gil et al. 1999; Saino

et al. 2002a). Adjustments in reproductive investment in response to mate attractiveness









were first tested by Burley in 1986 by manipulating the perceived attractiveness of a bi-

parental care species of zebra finch, Taeniopygia guttata, with colored leg bands. Burley

referred to this concept as the Differential Allocation Hypothesis (DAH).

The DAH is based on the premise that the attractiveness of a mate influences the

reproductive value of the breeding attempt with that mate (Sheldon 2000). Differences in

male trait expression will lead to selection acting on females to invest more in current

reproduction when the investment is balanced by a greater benefit. Attractive high quality

mates may affect the payoffs received from reproduction by providing greater direct and/

or indirect benefits to the female and her offspring than unattractive low quality mates.

However, greater investment in current reproduction comes at a cost by compromising

the amount of available resources for future reproduction. Therefore, selection for

differential allocation will depend on the magnitude of perceived benefits received from

reproduction. Furthermore, the expected future reproductive lifespan of the female and

the expected quality of future partners will also influence the role of differential

allocation. Increased allocation is only expected when the current partner is more

attractive than expected future partners (Sheldon 2000).

While attempting to explain the evolution of female mating preferences, models of

sexual selection have clearly described the potential benefits females receive through

mating. Indicator models propose that male traits evolve to "indicate" or "honestly

signal" some aspect of male phenotypic and/or genotypic quality (Zahavi 1975; Kodric-

Brown & Brown 1984; Grafen 1990). Male traits become targets of female preference as

a result of the correlation between the direct and/or indirect benefits a female receives

and the expression of the male trait. Indirect benefits, commonly referred to as "good









genes" are expected to increase offspring performance through inheritance of paternal

genes for viability (Norris 1993; Petrie 1994), sexual attractiveness (Gwinner & Schwabl

2005), and parasite resistance (Hamilton & Zuk 1982; Moller 1990). Alternatively,

females may directly benefit from non-heritable resources that increase female survival

or fecundity. Direct benefits include better parental care (Petrie 1983; Norris 1990),

courtship feeding (Nisbet 1973; Wiggins & Morris 1986), nuptial gifts (Thornhill 1976),

or access to resources such as superior nest and oviposition sites (Holm 1973;

Campanella & Wolf 1974).

For male traits to "honestly" represent the fitness benefits gained by females,

theory suggests that male traits must be physiologically costly either to produce or

maintain (Andersson 1994). Indicator models of sexual selection, therefore predict the

expression of the trait to be condition-dependent (Grafen 1990). That is, the degree of

trait expression is positively correlated with physical condition. Males in better condition

signal their quality through greater sexual trait size or more vigorous displays. Males in

worse condition incur higher costs that are associated with the production and/or

maintenance of these traits and are unable to express the traits to the magnitude that

males in better condition can. Evidence for honest advertisement has been found across

taxa (Johnstone 1995).

In practice, researchers have used measures of energy reserves (Bolger & Connolly

1989), vigor (Kodric-Brown & Nicoletto 1993), and/or health (Milinski & Bakker 1990)

to estimate an individual's condition. Empirical support for the influence of condition

measures on the expression of male traits, such as breeding coloration and courtship

rates, has been shown in fish. Research of guppies reveals that several of the traits which









females use to discriminate amongst males are condition-dependent and function to

indicate viability (Nicoletto 1993). For example, male display rate, amount of orange

coloration, and ranks of ornamental complexity were all found to be positively correlated

with condition measures (Nicoletto 1993). Breeding coloration has also been shown to be

positively correlated with condition measures in sticklebacks (Milinski & Bakker 1990)

and in pupfish (Kodric-Brown & Nicoletto 1993).

Furthermore, condition measures have been successful in explaining male variation

in reproductive success as a result of the influence of male energy reserves on parental

care. In the sand goby, Pomatoschistus minutus, food-supplemented males with higher fat

reserves spent more time at their nest caring for their young. As a result, these males

mated sooner and received more eggs than un-supplemented males (Lindstrom 1998).

Diet manipulation in small mouth bass also resulted in an increase in reproductive

success as a result of food-supplemented males providing longer periods of parental care

after swim up of larvae (Ridgway & Shuter 1994). Similar patterns of condition-

dependent parental care are also seen in the bicolor damselfish, Stegastespartitus, as

females receive higher quality parental care from more vigorously courting males which

is a reflection of male fat reserves (Knapp & Kovach 1991; Knapp 1995).

These experimental results suggest that condition-dependent expression of male

secondary sexual traits is common in many fish systems. Furthermore, female

preferences for condition-dependent traits often lead to greater direct and/or indirect

benefits. According to the DAH, selection should favor females who invest more with

these mates.









The objective of this study was to determine if female flagfish, differentially

allocate reproductive resources in response to male condition. Previous evidence in this

system suggests that females may discriminate amongst males based on condition. St.

Mary et al. (2001) showed condition values of flagfish males with low spawning success

to decline more drastically over the breeding season than more successful males.

Furthermore, evidence that females are evaluating males based on traits associated

with direct reproductive benefits makes flagfish an attractive system for the study of

differential allocation. Males provide exclusive parental care of eggs by defending

breeding territories from egg predators, oxygenating their developing eggs through

fanning, and cleaning their nests of debris. Parental care has been shown to be used in

mate attraction as a positive relationship was found between the numbers of eggs a male

received and initial fanning behavior (St. Mary et al. 2001). If the DAH holds, females

should allocate resources to more vigorously displaying males who may be in better

condition and are able to provide better parental care.

It is likely that parental care is a function of male energy reserves in this system.

Courtship takes place at the nest with males continually chasing and displaying towards

passing females. These activities confine males to their nests and may restrict foraging

opportunities to short periods of time interspersed between courting females and

aggressive interactions with neighboring males. Males may therefore be constrained by

food availability. This may affect their ability to provide parental care and subsequently

influence female resource allocation. Courtship displays and parental care are known to

be energetically costly in other fish species (Smith & Wootton 1999).









In this experiment, male condition was experimentally manipulated through diet by

assigning males to a high or low food treatment. This manipulation was expected to

affect measures of body size, fat reserves, color expression, and courtship/parental care

behaviors. Over a two week period, females interacted sequentially with two males from

equal or opposite feeding treatments. To evaluate differential allocation the total number

of eggs spawned with each male was recorded in addition to measures of hatching

success, egg energy, egg size, and larval length. Under the DAH, females were expected

to provide a larger number of eggs and/or higher quality eggs to males who have been

maintained on high food rations than low food rations.















METHODS

Study Species

Flagfish, Jordanellafloridae, are found in fresh, brackish and salt water systems of

central and southern Florida (Foster et al. 1969). Flagfish inhabit areas of dense

vegetation and live approximately one year (Boschung et al. 2000). Flagfish have a

promiscuous mating system. Their breeding season extends from late March to early

September (Boschung et al. 2000). Males defend breeding territories from male

competitors and egg predators. During courtship, males will repeatedly chase and circle

the female. Spawning occurs on submerged vegetation or leaf litter within the breeding

territory. During daylight hours, a female may spawn with a single male multiple times or

distribute her eggs among males (Mertz & Barlow 1966). Females will lay one egg at a

time, releasing approximately 20 or more eggs during a single spawning bout. Males

continually accept eggs from multiple females and provide parental care by fanning,

guarding, and cleaning their eggs (Mertz & Barlow 1966). Males continue to care for

their eggs until hatching occurs within 4-5 days of spawning at 250C (Smith 1973).

Flagfish show sexual dimorphism in both body size and coloration. Males are

larger in size and are the more colorful sex. Males possess horizontal red-orange stripes

on their dorsal and anal fins as well as along their side. A large dark spot surrounded by

yellow is located on their mid side and blue iridescence appears on their operculum.

During periods of courtship and nest guarding, males temporarily express dark coloration









over their entire body. Females are silvery gray in color and are characterized by a dark

spot located at the tail end of their dorsal fin (Page & Burr 1991).

Study Design

This study was conducted at the University of Florida in Gainesville, Florida.

Experimental research was carried out over one breeding season from March to August

of 2005. Flagfish for this study were collected during the summer of 2005 from the Otter

Creek/Waccasassa River drainage system in northwest central Florida. Sexes were

housed separately in 152 liter holding tanks and maintained on a 14L:10D light cycle.

The room was kept at a minimum of 270 C.

Experimental Manipulation of Male Condition

Males were randomly assigned to one of two food treatments. In the holding tanks,

high food (HF) males were fed a diet of algae tabs, commercial flake food, and frozen

chironomid larvae in excess two times a day. Low food males (LF) received a diet of

algae tabs and were fed once every other day. Males were maintained on the diet for a

period of 14 to 42 days prior to interacting with a given female. Females were fed in the

same manner as high food males in order to maximize their potential for egg production.

To control the diet of both sexes during female mating trials, males and females were

separated by a clear acrylic partition and fed independently at the end of each day. Any

remaining food was removed from the tank before the start of the mating trial the

following morning using a fine mesh net.

Female Mating Trials

A single female and single male were placed in a 38 liter holding tank. A spawning

mat was set in the center of the tank and an artificial plant (Ludwigia species) was

positioned in the back of the tank. The spawning mat was constructed from a square









ceramic tile (10cm x 10cm) with green felt carpet attached to the top. Male flagfish have

been observed in previous research to readily accept these spawning mats as breeding

territories and typically all eggs are spawned on the mat. Clear monofilament fishing line

was used to separate the spawning mat into 20 square quadrants to aid in the task of

counting freshly spawned eggs.

Over a ten day period, each female was given the opportunity to interact with two

different males for five days each. Each male-female pair interacted from 0830 to 1630

hours, for a total of eight hours every day. At the end of each day a perforated clear

acrylic partition was placed in the center of the tank between the male and female to

prevent spawning overnight. Clear perforated partitions were used to facilitate visual and

chemical communication between the pair throughout the night. At the end of the first

five days the male was removed and replaced with a new male from the opposite or same

feeding treatment. Each female was randomly assigned to one of the following four

treatments; 1) HF/HF 2) LF/LF 3) HF/LF and 4) LF/ HF. For every ten day period, each

treatment was replicated four times. This design ensured that the treatments were evenly

distributed throughout the breeding season. Each male and female was used only once.

Prior to each five day mating trial, perforated clear acrylic partitions were used to

physically separate each male-female pair for a two day period. This period of

acclimation gave each male a sufficient amount of time to establish his breeding territory

over the spawning mat. Prior to interacting in the mating trial, the fish were weighed on

an electronic balance to the nearest 0. g and measured for standard length to the nearest

0.lcm.









Measures of Reproductive Allocation

To assess differential allocation the following five parameters were measured with

each male 1) total number of eggs spawned with each male 2) average individual egg size

per clutch 3) average egg energy content per clutch 4) hatching success rate of larvae and

5) average larval length per clutch.

Spawning mats were checked for eggs four times a day at approximately 0900

hours, 1100 hours, 1300 hours and 1600 hours. At each time the total number of new

eggs was recorded. Freshly spawned eggs were differentiated from older eggs by viewing

the developmental stage under a dissecting scope and by noting the placement of eggs

within the monofilament grid. After the first spawning event, up to 30 eggs were removed

from the spawning mat. The remaining eggs were left on the mat to promote continued

spawning. If the removal of 30 eggs reduced the total clutch size by more than fifty

percent, eggs were removed on subsequent spawning events. Of these eggs, 15 were

immediately frozen for energetic analysis. The remaining 15 eggs were then placed in a

water filled plastic dish and digitally photographed under a dissecting scope.

ImageProPlus (Version 4.5.1) software was used to measure the total area (mm2) of each

individual egg within a clutch. To reduce error, three area measurements were recorded

and an average of these measurements was used in calculating the average egg area for

the entire clutch. The eggs were then placed in a 100 ml water filled glass rearing

chamber. Each rearing chamber was supplied with an air stone and maintained at room

temperature of 250C. Hatching success was measured as the proportion of fertilized eggs

hatched by day seven. Once hatched, larvae were digitally photographed by the same

methods used for the eggs. Three measurements of total length (mm) were taken for each









larvae and an average of these measurements was used in calculating the average larval

length for the entire clutch.

To measure the average egg energy content per clutch a dichromate oxidation

method was used following the procedure of McEdward and Coulter (1987). Three eggs

were randomly chosen from the 15 egg sample. Each egg was incubated for 15 minutes at

1100 C in lml of 70% phosphoric acid and then oxidized in 2 ml of 0.30% potassium

dichromate for an additional 15 minutes and incubated at the same temperature. Samples

were diluted with 3.5ml distilled water and energy was measured with a

spectrophotometer (440nm) by comparison of the sample to glucose standards ranging

from 0 to 4 joules. An average of the three measurements was calculated to estimate the

egg energy content per clutch.

Color Analysis of Males

To record variation in color expression between males, digital photographs of each

male were taken with an Olympus C 2500L camera. To ensure the development of

nuptial coloration, photographs were taken at the end of the first day of each five day

mating trial in order to allow males significant interaction time with females. Each male

was hand-netted and placed in a clear water filled acrylic box (9cm x 3cm x 6cm) within

a standardized photo-box. The small box prevented the movement of the fish, exposing

the lateral side of the male to the camera. Each photo included a red, yellow, and blue

color standard. For each photograph we first sampled the three color standards recording

the hue, saturation, and intensity of each. This allowed for the adjustment of variation in

light intensity between images when recording color measurements. Red was defined as

having hue values ranging from 0 to 40 and saturation values ranging from 30 to 100;

yellow was defined as hue values from 40 to 55 and saturation values from 70 to 100; and









blue was defined as hue values from 100 to 160 and saturation values from 25 to 100. An

image analysis software tool, SigmaScanPro (Version 5.0.0), was used to measure the

mean intensity of red, yellow and blue coloration for each male. The total area of each

color on the fish's body was also measured and a proportion was calculated from the area

of the entire fish.

Lipid Analysis of Males

A sample of males from each food treatment were randomly selected and

euthenized in MS-222 at the end of the five day female mating trial. Males were frozen

and then later dried in an oven at 600 C for 24 hours. Upon removal, the dried males were

weighed on an electronic balance to the nearest 0.0001g. To assess differences in male

condition as a result of diet manipulation, lipids were extracted with petroleum ether

(Knapp 1995). Males were continually placed in ether until a constant dry weight was

achieved. The difference in weight was then calculated and the percentage of fat weight/

dry weight was used as a measure of condition in data analyses. Fresh weight/length3 was

also used as a second condition measure.

Behavioral Observations of Males

Males were videotaped using a VHS recorder for ten minutes on days one and two

of the five day mating trial between 1200 hours and 1600 hours. Flagfish are more active

in the afternoon (personal observation). This provided an estimate of a male's

reproductive behavior. To evaluate parental care, males were also taped for an additional

ten minutes in the presence of their first bout of eggs. Male behavior was analyzed using

an event recorder program written for this purpose. Male position was recorded as 1) at

the nest or 2) away from the nest. Measured male behaviors included 1) nest fanning 2)

nest cleaning 3) following 4) chasing and 5) spawning (see Table 1 for a more detailed









descriptions of these behaviors). Chasing and cleaning were recorded as counts since the

duration of these behaviors is short. Duration was recorded for fanning, following, and

spawning. The percentage of total observation time spent at the nest, away from the nest,

following, and spawning was calculated for each male. Frequencies for count behaviors

were calculated as the number of times a behavior was performed divided by the total

observation time. Fanning and cleaning are behaviors that occur only while the male is at

the nest. Therefore the percentage of time fanning and the frequency of cleaning were

calculated from the total time spent at the nest rather than the total ten minute observation

time.

Statistical Analysis

For all analyses, only data from females (and their mates) that spawned in at least

one week of the two week mating trial was considered. Parametric statistical analyses

were utilized when the variables met the requirements of these methods. For all variables

normality was tested using the one-sample Kolmogorov-Smirnov Test. If the variables

did not meet normality requirements, the appropriate transformations were made.

Variables expressed as proportions were arc sin square-root transformed. These included

color proportions as well as the proportion of measured fat weight/dry weight (g). The

numbers of eggs a female spawned were square-root transformed. Male and female fresh

weights (g) and standard lengths (cm) were log transformed. For general linear models,

variables with p-values greater than 0.15 were removed in a step wise fashion until the p-

values that remained were less than this set criterion. Nonparametric statistics were used

for behavior data since these variables did not satisfy normality assumptions when

transformed. All statistical analyses were performed using SPSS Version 12.















RESULTS

Effects of Diet Manipulation on Male Body Size, Condition, Coloration & Behavior

Male food treatment was found to have a significant effect on male body size and

coloration, though no differences were found for measures of male condition, fat

reserves, or behavior. High food males on average were significantly heavier and longer

than low food males (Table 2). However when controlling for length, condition did not

differ. To evaluate the effect of food treatment on male condition, a multivariate analysis

of variance was performed on fresh weight, dry weight, and fat weight with male

standard length as a covariate. All weight variables increased with standard length for

both food groups as expected (Figuresl-3 MANCOVA Fresh Weight F1, 124 = 997.61 P =

0.00; Dry Weight F1, 124 = 473.47. P = 0.00; Fat Weight F1, 124 = 98.78 P = 0.00). Male

food treatment had no significant effect (MANCOVA F3, 122 = 1.33 P = 0.27 Wilk's k =

0.97). Furthermore, weight measures did not increase more sharply with body length for

high food males than low food males (MANCOVA Food Treatment* Standard Length F3,

122 = 1.36 P = 0.26 Wilk's X = 0.97). Thus, while diet manipulation did have a significant

effect on the weight and standard length of high food males, their body composition

remained stable. Comparisons of % fat weight/dry weight showed no significant

differences in fat reserves between male food treatments (Table 2).

High food males expressed a significantly larger proportion of blue coloration as

well as more intense yellow coloration than low food males (Table 2). No differences









were seen between male food treatments for the proportion of red and yellow coloration

or the intensities of red and blue coloration (Table 2).

Although high food males were expected to display more vigorously, no difference

was found in the amount of time spent in courtship behavior between male food

treatments (Table 3). Furthermore, the probability that high food males performed each

measured behavior during courtship was not significantly different from that of low food

males (Table 3). Behavioral differences were also expected post-spawning. Male food

treatment did not influence the amount of time spent in each behavior once eggs were

received (Kruskal-Wallis Test At Nest X2 = 0.78 P = 0.34; Fanning At Nest X2 = 1.41 P =

0.24; Chase Frequency X2 = 0.85 P = 0.34, Following X2 = 0.37 P = 0.54; Cleaning

Frequency X2 = 0.11 P = 0.75).

Measures of Reproductive Allocation & Male Food Treatment

Females were expected to alter their reproductive investment in response to male

food treatment by allocating a larger number of eggs to high food males. These males

were also expected to receive clutches with larger more energetic eggs, a higher hatching

success rate and larger larvae. No such patterns were found.

To determine female egg allocation patterns with respect to male food treatment, a

repeated measures analysis of variance was used to investigate statistical differences in

egg numbers between weeks one and two of the mating trial. Two separate analyses were

completed, one in which females were paired with males from the same food treatment

(LF/LF & HF/HF) and a second in which females were matched with males from

separate food treatments (LF/HF & HF/LF). The first analysis determined if females on

average spawned more eggs when two high food males were received. The second

analysis determined if the order of male presentation affected female egg allocation.









Females paired with high food males during both weeks one and two, did not

allocate more eggs than females who received only low food males. Female treatment

had no significant effect on the average number of eggs spawned over weeks one and two

of the mating trial (LF/LF & HF/HF F1, 36 = 1.92 P = 0.18). Furthermore, the difference in

eggs spawned between weeks one and two did not vary with female treatment. There was

no significant interaction between week and female treatment when considering females

matched with males from the same food treatment (LF/LF & HF/HF Week*Treatment F1,

36 = 0.00 P = 0.95). The order in which high food males were presented had no affect on

patterns of egg allocation given a non-significant interaction between week and treatment

when considering females matched with males from separate food treatments (HF/LF &

LF/HF Week*Treatment F1, 50= 0.71 P = 0.40).

Females were not limited in their ability to allocate eggs between weeks. That is

females that spawn in week one are not more or less likely to spawn in week two. Egg

number did not differ between week one and two of the experiment across female

treatments (LF/LF & HF/HF F1, 36 = 0.026 P = 0.872; HF/LF & LF/HF F1, 50 = 0.261 P =

0.612). Given the lack of significant week effects, differences in egg allocation between

high food and low food males were directly analyzed in a third repeated measures

analysis. In this case, week was ignored, and differences between high food eggs and

corresponding low food eggs were compared. Within females there was no significant

difference in the number of eggs spawned with a high food versus a low food male (F1, 50

= 0.15 P = 0.70). Hence, no apparent patterns were seen in female spawning between

male food treatments.









Considering clutches from both weeks one and two, male treatment did not affect

hatching success (Table 4). Furthermore, when considering only the HF/LF treatment,

hatching success did not differ between male food treatments (Paired T-test t = -0.30, df=

7 P = 0.78). Low sample size prevented this same analysis with the LF/HF female

treatment. Overall, hatching success rate was high for both male food treatments.

Across all female treatments, there was a significant difference in egg size between

weeks for females that spawned both weeks of the mating trial (Paired T-test t = 2.99 df=

13 P = 0.01). Females spawned larger eggs in week one (1.15mm2 0.03 SE) compared

to week two (1.08mm2 0.02 SE) suggesting that females are limited in producing larger

eggs as spawning continues. Given this result, eggs produced during the second week of

the mating trial were removed from further analyses of egg size, egg energy and larval

length if the female spawned both weeks. Male food treatment did not affect average

individual egg size, average egg energy or average larval length (Table 4).

Measures of Reproductive Allocation & Male Traits

Additional analyses were utilized to investigate the association between allocation

variables and male morphological and/or behavioral traits. Variation in egg number, size,

and energy among females can not be attributed to morphological and/or behavioral

aspects of the sire. Larval length shows an exception.

The relationship between differences in egg numbers between week one and two of

the experiment with differences in male morphology and behavior between mates was

evaluated. A linear relationship was expected, with females allocating more eggs to males

with greater trait expression. Differences in egg numbers did not co-vary with differences

in male body size, condition, fat reserves and coloration (ANCOVA Weight F1, 40= 0.02

P = 0.90; Standard Length F1, 75 = 1.20 P = 0.28; Weight/Length3 F1,74 = 1.16 P = 0.29; %









Fat F1,45 = 0.13 P = 0.72; % Blue F1,43 = 0.08 P = 0.77; Intensity Blue F1,76 = 1.37 P =

0.25; % Red F1,41 = 0.04 P = 0.84; Intensity Red F1,42 = 0.05 P = 0.83; % Yellow F1,73

0.02 P = 0.89; Intensity Yellow F1,77= 2.20 P = 0.14). Nor were any correlations seen

with variation in male behavior (At Nest Spearman's p = 0.02 P = 0.85; Fanning At Nest

Spearman's p = 0.03 P =0.79; Cleaning Frequency Spearman's p = -0.02 P = 0.90;

Chasing Frequency Spearman's p = 0.03 P = 0.80; Following Spearman's p = -0.08 P =

0.53).

To determine if females respond to differences in male morphology when

allocating eggs of varying size and energy, a univariate analysis of covariance was used.

Measures of color and male and female condition were used as covariates with male

treatment as a fixed factor. Male food treatment did not affect egg size or energy as

expected from previous results (ANCOVA Egg Size F1, 40 = 0.18 P = 0.68; Egg energy

F1, 34 = 1.75 P = 0.20). Male and female condition and blue and red coloration were not

found to co-vary with egg size or energy, nor were any significant correlations found

between egg size and energy and measures of yellow coloration or aspects of male

courtship behavior prior to spawning (Table 5). Male condition and red coloration did not

co-vary with egg size however marginal p-values were found. Therefore, residuals from

the ANCOVA model with proportion red as a covariate were regressed against male

condition. The linear regression was non-significant (R2 = 0.05 F1, 60 = 3.25 P = 0.08)

however a negative relationship between the variables are seen (Figure 4).

Similar analyses to those conducted on egg size and energy were utilized for larval

length. Measures of average hatch date, color, and male and female condition were used

as covariates with male treatment as a fixed factor. Average hatch date had a significant









positive influence on average larval length (ANCOVA F1,52 = 16.96 P = 0.00; Figure 5

Linear Regression R2 = 0.22 F1,79 = 22.63 P = 0.00). There was no effect of male food

treatment (ANCOVA F1, 48 = 0.16 P = 0.69) or female condition on average larval length

(Table 5). However, male condition showed significant effects on average larval length

(ANCOVA F1, 52 = 9.74, P = 0.00.) Residuals from an analysis of variance with average

hatch date as a covariate and average larval length as the response were regressed against

male condition. A negative relationship was found between average larval length and

male condition (Figure 6 Linear Regression R2 = 0.15 F1,65 = 11.64 P = 0.00) similar to

that found with egg size analyses. These results suggest that males of a lower weight for a

given length father larger larvae and received eggs of a greater size. Blue and red

coloration were not found to co-vary with larval length nor were any significant

correlations found between the residuals from the above analysis and measures of yellow

coloration or aspects of male courtship behavior prior to spawning (Table 5).

When hatch date is taken into consideration, egg area has a significant effect on

larval length (ANCOVA Average Hatch Date F1,67 = 11.50 P = 0.00; Average Egg Area

F1, 67= 5.06 P = 0.03). Residuals from an analysis of variance of larval length with

average hatch date as a covariate were regressed against average egg area. This

regression demonstrates a positive relationship between average larval length and average

egg area (Figure 7 Linear Regression R2 = 0.07 F1, 68 = 4.89 P = 0.03). Therefore, egg

size is predictive of larval length.

Female Mate Choice

High food males were expected to obtain more mates given the prediction that

these males would be in better condition. Females did not favor mating with one male

food treatment over the other. In week one of the mating trial, high food males were









successful in spawning 70% of the time in comparison to low food males with 60%

During week one, females were not more likely to mate with high food males than low

food males (Chi-square X2 = 1.09 P > 0.1). The same results are found when considering

week two. The probability of mating with high food verses low food males in week two

was not significantly different (Chi-square X2 0.29, v =1, P > 0.5). High food males had

a spawning success rate of 73% while low food males showed similar success with 78%.

On average, mated males were not significantly heavier or longer than unmated

males, nor were they in better condition, had greater fat reserves or expressed more

coloration (Table 6). Results of behavior analyses do indicate however, that female

flagfish prefer to mate with males who exhibit signs of parental care during courtship.

Prior to receiving eggs, mated males on average spent more time defending and fanning

their nests (Figures 8-9 Kruskal-Wallis Test At Nest X2 = 7.29 P = 0.01; Fanning At Nest

X = 5.08 P = 0.02). In addition, mated males chased females more frequently (Figure 10

Kruskal-Wallis Test X2 = 7.19, P = 0.01) than unmated males. Furthermore, the

probability that mated males performed these behaviors was greater than that of unmated

males (Chi-Square At Nest X2 = 4.18 P < 0.05; Fanning At Nest X2 =12.18 P < 0.001;

Chase Frequency X2 =7.59 P < 0.5). The proportion of time spent following a female and

the frequency of cleaning events did not differ between groups (Kruskal-Wallis Test

Following X2 = 0.73 P = 0.39; Cleaning Frequency X2 = 1.93 P = 0.16) nor did the

probability with which these behaviors were performed (Chi-Square Following X2 =0.79

P > 0.05; Cleaning Frequency X2 =2.35 P > 0.5). These results are consistent with

previous studies which demonstrate that female flagfish show preferences for increased

parental care during courtship (St. Mary & Lindstrom in prep; Hale & St. Mary in prep.).









The amount of time males spend fanning during courtship provides females with an

a priori expectation of the quality of parental care their offspring will receive. An

analysis of covariance, shows a significant influence of egg number (F1,60 = 23.76 P =

0.000) and time spent fanning pre-spawning (F1,60 = 9.20 P = 0.00) on fanning at the nest

post-spawning. A regression of the residuals from an analysis of covariance with fanning

post-spawning as the dependent variable and egg number as a covariate, indicates a

significant relationship between fanning pre-spawning and fanning-post spawning (R2

0.13 F1,61 = 9.45 P = 0.00), suggesting that mates who fan during courtship are also

likely to exhibit the behavior after spawning.

Condition-Dependent Expression of Male Traits

Diet manipulation was expected to generate differences in male condition. As a

result, males were expected to show condition-dependent expression of potentially costly

traits such as courtship behavior and coloration. Of the behaviors measured, fanning the

nest and chasing the female possibly require the greatest amount of energy from the male.

Measures of male condition and fat reserves were unable to explain the variation in the

frequency of chases a male performed during courtship (Table 7). However, a negative

relationship between fanning effort and fat reserves was found when considering only

those males who spent time fanning their nests (Figure 11 Linear Regression % Fat

Weight F 1, 37 = 3.96 P = 0.05). This relationship was not seen however when considering

an additional measure of condition (Linear Regression Weight/Length3 F1, 51 = 0.29 P =

0.60). Measures of male condition and fat reserves were not predictive of blue and red

color expression, nor were they correlated with yellow color expression (Table 7)









Table 1. Description of nest and non-nest directed behaviors recorded for each male
flagfish.


Behavior


Position

At the Nest


Away From Nest


Description


Located within 6cm of vertical
distance above the nest with part
of the body over the nest.

Located away from the nest.


Nest Directed Behavior


Fanning


Body is angled downward toward
nest while performing a rapid
side-to-side movement.


Cleaning

Spawning


A bite at the nest.


Synchronous movement of fish
towards nests. Fish are paired side
by side in close proximity, releasing
eggs as they make direct contact
with the nest.


Non Nest Directed Behavior


Following

Chasing


Following of female.


Rapid swimming directed towards
female









Table 2.The effect of male food treatment on morphological traits.

High Food Low Food Test P-
Average ( SE) Average ( SE) Statistic Value


Measures of Body Size
and Condition


Fresh Weight (g) 1.97 (0.09) 1.69 (0.08) 5.727 0.02*

Standard Length (cm) 3.64 (0.06) 3.46 (0.05) 4.685 0.03*

% Fat Weight / Dry 0.19 (0.01) 0.21 (0.01) 0.521 0.47*
Weight

Measures of
Coloration


% Body Area Blue 8.56 (0.00) 5.06 (0.00) 6.40 0.01 *

% Body Area Red 0.04 (0.00) 0.05 (0.00) 0.28 0.60 *

% Body Area Yellow 0.00 (0.00) 0.00 (0.00) 0.74 0.39 t

Intensity Blue 3.54 (0.14) 3.50 (0.16) 0.03 0.86*

Intensity Red 2.36 (0.05) 2.29 (0.05) 0.43 0.51 *

Intensity Yellow 1.33 (0.10) 1.02 (0.11) 5.07 0.02 t

*ANOVA (two-tailed) with F Test Statistic
t Kruskal- Wallis Test with X2 Test Statistic









Table 3. The effect of male food treatment on measures of courtship prior to spawning.
Statistical analyses were used to determine differences between high and low
food males for 1) the % of observation time a behavior was performed
(Kruskal-Wallis Test) 2) the frequency with which a behavior was performed
(Kruskal-Wallis Test) and 3) the probability that a behavior was performed
(Chi-Square).


Measures of
Male Behavior


Kruskal-Wallis


Test Statistic P- Value


Chi-Square

Test Statistic P- Value


% Time

At Nest
Fanning At Nest
Following


Frequency

Cleaning
Chasing


1.85
0.09
2.18


1.85
0.34


0.17
0.76
0.14


0.17
0.56


1.04
1.99
2.04


0.06
0.04


>0.1
>0.1
>0.1


> 0.5
>0.5









Table 4. The effect of male food treatment on female reproductive allocation. Measures
include hatching success, average egg size, average larval length, and average
egg energetic content.


Measure of
Reproductive Output


% Hatched Larvae

Egg Area (mm2)

Larval Length (mm)

Egg Energy (J)


High Food
Average ( SE)


86.91 (0.03)

1.13 (0.02)

3.88 (0.06)

1.88 (0.12)


Low Food
Average ( SE)


88.90 (0.03)

1.12 (0.01)

3.96 (0.05)

1.70 (0.12)


*ANOVA (two-tailed) with F Test Statistic
t Kruskal- Wallis Test with X2 Test Statistic


Test
Statistic


0.12

0.14

0.96

1.10


P- Value



0.72t

0.71*

0.33*

0.30*









Table 5. Results of analyses addressing variation in average egg area and larval length in
response to male morphological and behavioral traits.

Egg Larval Egg
Area (mm) Length (mm) Energetics (J)

Test P Test P Test P
Statistic Value Statistic Value Statistic Value


Measures of
Condition

Male Weight/ 3.14
Length3

FemaleWeight / 0.28
Length3

Measures of
Coloration

% Body Area Blue 1.79

% Body Area Red 3.28

% Body Area 0.24
Yellow

Intensity Blue 0.22

Intensity Red 0.10

Intensity Yellow 0.10

Measures of
Behavior

At Nest 0.03

Fanning At Nest -0.25

Cleaning 0.19
Frequency
* ANCOVA with F Test Statistic
8 Spearman's Rank Nonparametric


0.08


0.60






0.19

0.08

0.10


0.64

0.75

0.51




0.81

0.16

0.30


9.74


0.16






3.29

0.91

-0.14


1.52

1.61

-0.23




0.02

-0.02

-0.04


0.00* 0.59 0.45


0.69* 2.97 0.09


0.08*

0.34*

0.314


0.22*

0.21*

0.081





0.87

0.935

0.84t


0.29

0.01

-0.02


1.50

0.44

-0.09




-0.01

0.02

-0.03


0.59

0.91

0.901


0.23

0.51

0.621





0.951

0.951

0.921


Correlation (two-tailed) with Correlation Coefficient









Table 6. Differences between mated and unmated males in measures of male body size,
fat reserves, condition factor, and coloration.

Mated Unmated Test P- Value
Average ( SE) Average ( SE) Statistic


Measures of Body Size
and Condition


Fresh Weight (g) 1.81 (0.08) 1.89 (0.11) 0.77 0.38*

Standard Length (cm) 3.54 (0.05) 3.58 (0.07) 0.25 0.62*

% Fat Weight / Dry 1.87 0.17*
Weight 0.19 (0.00) 0.21 (0.00)

Weight/ Length3 0.04 (0.00) 0.04 (0.00) 1.78 0.18*

Measures of
Coloration


% Body Area Blue 0.04 (0.02) 0.07 (0.03) 0.00 0.99*

% Body Area Red 0.04 (0.01) 0.05 (0.02) 1.31 0.25t

% Body Area Yellow 0.00 (0.00) 0.00 (0.00) 1.56 0.21*

Intensity Blue 3.52 (0.12) 3.51 (0.20) 0.00 0.96*

Intensity Red 2.32 (0.07) 2.34 (0.07) 0.06 0.81t

Intensity Yellow 1.24 (0.14) 1.32 (0.14) 0.51 0.47*

*ANOVA (two-tailed) with F Test Statistic
t Kruskal- Wallis Test with X2 Test Statistic









Table 7. Results of condition-dependent expression analyses.

Weight / Length3 % Fat Weight / Dry Weight

Test Statistic P-Value Test Statistic P-Value


Measures of Coloration


% Body Area Blue 0.07 0.79 0.08 0.78

% Body Area Red 3.01 0.08 0.06 0.80

% Body Area Yellow -0.07 0.38 -0.01 0.90O

Intensity Blue 0.15 0.70 0.08 0.30

Intensity Red 0.15 0.70 0.85 0.36

Intensity Yellow -0.07 0.37 -0.09 0.36

Measures of Behavior


Fanning At Nest 0.29 0.60 3.96 0.05

Chasing Frequency 0.80 0.38 2.38 0.13

% Spearman's Rank Nonparametric Correlation (two-tailed) with Correlation Coefficient
"Linear Regression with F Test Statistic











O High Food
A Low Food


6.00-



5.00-



^4.00-

300
0)


2.00-
-C

2.00-



1.00-



0.00-


I I I I I
2.00 3.00 4.00 5.00 6.00
Standard Length (cm)


Figure 1. The relationship between fresh weight and standard length is shown for both
high food and low food males.


0
A O
O
O
8R












0
0


O
A

o O
AO


0 High Food
A Low Food


I I I I I
2.00 3.00 4.00 5.00 6.00
Standard Length (cm)

Figure 2. The relationship between dry weight and standard length is shown for both high
food and low food males.


1.40-


1.20-


1.00-

0)
S0.80-
03)
-


:0.60-
-


0.40-


0.20-


0.00-












0 High Food
A Low Food


0.50-




0.40-




20.30-
-
0)


2 0.20-




0.10-




0.00-


0
A A




Mt A'- AO 0


I I I I I
2.00 3.00 4.00 5.00 6.00
Standard Length (cm)

Figure 3. The relationship between fat weight and standard length is shown for both high
food and low food males.


0 0
A











0.20-
-


E
S0.10-



LU
c)




0
S0.00-

0)


-0.10-

ry



-0.20-


R2 = 0.05


0
00


0 0


0 0 0 -
0
0% 0 00 0


0 @ 00 0
0 0 0
0 0o o
O0
O
0o


I
0.03


I
0.04


I
0.04


I
0.04


I
0.05


Male Condition (Fresh Weight (g) / Standard Length (cm))3


Figure 4. Values of male condition are plotted against residuals from a univariate analysis
of co-variance in which average egg area was used as the dependent variable
and % red coloration of the sire as a covariate .














4.60-


4.40-


4.20-


4.00-


3.80-


3.60-


3.40-


3.20-


R2 = 0.22


0




o 0o
O,


0
0


0 o
O
80
0


I I I I I I I
3.00 3.50 4.00 4.50 5.00 5.50 6.00

Average Hatch Date (Days)


Figure 5. Average larval length (mm), calculated as the average total length of individual
larvae within a clutch is plotted against the average hatch date for that clutch.
Average hatch date was calculated as the sum of the # of individual larvae
times their respective hatch dates divided by the total number of hatched
larvae.











0.75- R2 = 0.15

O O
E 0.50-
E 0 0
O

O OO


0 o
> o o
-, 0 0 0







a, o o8; o
c,)
(D 0.25- o 0
-FU








(S0 0 0
00




-0.50- o
(D 0
> o 0 0O0
< -0.25- o


'0 0
-0.50-



-0.75 0


0.03 0.04 0.04 0.04 0.05

Male Condition (Fresh Weight (g) / Standard Length (cm) )3

Figure 6. Values of male condition are plotted against residuals from a univariate analysis
of co-variance in which average larval length was used as the dependent
variable and average hatch date as a covariate.











0.75-


0.50-



0.25-



0.00-



-0.25-



-0.50-


-0.75-


I
0.90


R2= 0.07


0
0 0


000
0
O




0 000
oo




0 0 0
o' o o


0
o


I
1.00


O o


C 0
9 0
0(9


I
1.10


I
1.20


I
1.30


I
1.40


Average Egg Area (mm2)
Figure 7. Individual egg areas were measured within a clutch and then averaged. These
values are plotted against residuals from a univariate analysis of co- variance
in which average larval length was used as the dependent variable and average
hatch date as a covariate.



















(D 0.30
z





0.20



O
0
0

o 0.10





0.00
Mated Unmated


Figure 8. Average proportion of total observation time each male spent at their nest(
SE) for mated and unmated males prior to receiving eggs.















0.04
0)


U-
0.03




E
iF 0.02
4-
0



0.01




0.00
Mated Unmated


Figure 9. Average proportion of time spent at the nest fanning (+ SE) for mated and
unmated males prior to receiving eggs.













0.01



0.01 -
















Mated Unmated

Figure 10. Frequency of chases ( SE) performed by mated and unmated males during
their recorded observation time prior to receiving eggs. Frequency of chases
was calculated as the number of chases performed divided by the total
observation time.0.01
u.







0.00




Mated Unmated

Figure 10. Frequency of chases (+ SE) performed by mated and unmated males during
their recorded observation time prior to receiving eggs. Frequency of chases
was calculated as the number of chases performed divided by the total
observation time.












0.70- R2 = 0.10
0

0

0.60- 8

)m 0

S0.50- 0 o o
uo ^ o o o o
LL

00
S0.40 o
< o
E o
4- o o 0 o
- 0.30-

0 0 0

0.20-
o o

o @
0.10- o


0.00 0.10 0.20 0.30 0.40 0.50 0.60

% Fat Weight (g) / Dry Weight (g)


Figure 11. Relationship between fanning and fat reserves. Transformed values are show.















DISCUSSION

The current study attempted to influence male condition through diet. This

manipulation was expected to affect measures of body size as well as fat reserves. Male

food treatment had a significant effect on male weight and standard length, as also

previously found when a comparable HF/LF diet was implemented (Klug & St. Mary

2005). However, diet manipulation did not influence measures of condition. High food

males maintained their body composition, as weight did not increase more sharply with

body length. Additional nutrients were utilized for growth as opposed to accumulating in

stored fat reserves. This may suggest that body size plays a valuable role in the flagfish

mating system.

Males may prioritize energy allocation to increased size with the aim of acquiring

and defending breeding territories essential for reproductive success. In nature flagfish

often aggregate and reproductive sites often occur in close proximity (R Hale personal

observation). Hence, high density conditions are likely. Larger males will be more

effective at excluding potential competitors and thus able to spawn more without

disruption, resulting in selection for males to invest in growth. Additionally, female

preference for larger dominant males may further enhance these selective pressures.

In the current study, variation in egg number among males was not related to male

size. Nor were mated males found to be of larger size. However, these results may be

influenced by the single male presentation. Female preferences in other species have been

shown to change in response to their social environment (Kvarnemo et al. 1995; Forsgren









et al. 1996; Kangas & Lindstrom 2001). St. Mary and Lindstrom (unpublished data)

suggest a similar response in flagfish as male traits associated with competitive ability,

such as body size and aggressiveness (frequency of chases towards males) became more

important in mating and egg allocation decisions as the level of male-male competition

increased from the absence of competition (single male and single female) to moderate

(two males and two females) and high (four males and one female) levels.

Males were predicted to show condition-dependent expression of the traits

potentially used in mate choice. Measures of color expression were expected to be

positively correlated with a male's physical condition. Of the three colors measured, red

and yellow were specifically expected to be honest indicators of condition as these have

been demonstrated to be carotenoid based in several fish systems (Kodric-Brown 1998).

Since carotenoids cannot be synthesized, they must be acquired through the diet (Olson &

Owens 1998). As sources are scarce in nature, the intensity of carotenoid based

coloration should reflect the content of a male's diet and his overall foraging ability

(Kodric-Brown 1989). In addition, carotenoids function to support the immune system.

Recruitment of carotenoids to non-recoverable forms in the dead tissue of scales may

compromise a male's overall health, increasing his susceptibility to pathogens or

parasites (Folstad & Karter 1992). Only males in superior condition should be able to

meet this expense. Carotenoid expression should therefore provide a dependable basis for

females to choose males in good condition (Nicoletto 1993).

In the current study, measures of male condition were unable to explain the

variation in color that existed between males. However, diet manipulation was successful

in influencing aspects of color expression. Diet influenced the proportion of the body









covered with blue coloration as well as the intensity of yellow coloration, as high food

males showed greater expression in each case. While, reds, oranges, and yellows are

generally thought to represent carotenoid expression, pale purples, greens and blues can

be produced when carotenoids are bound to proteins (Olson & Owens 1998.) Algae are

the primary water-based sources of carotenoids for fish, in addition to aquatic

invertebrates (Olson & Owens 1998). High food males were fed these sources in

abundance, suggesting that blue pigments may be carotenoid based. Alternatively, blue

coloration has been found to be mediated by the expansion and contraction of

melanophores in other fish systems (Kodric-Brown 1996). Measures of red coloration

were not affected by diet manipulation, suggesting that these pigments may originate

from other sources. Red, yellow, and orange coloration are also produced by pteridine

pigments derived from purines which are synthesized from carbohydrates and proteins

(Hurst 1980). Pteridines have been found to contribute to sexual coloration in poeciliid

fishes as orange spots of male guppies were found to contain red pteridine pigments in

addition to carotenoids (Grether et al. 2001).

In light of these results, it seems that color expression may well be diet dependent.

However, it is not a simple reflection of fat reserves for instance. For example, previous

research in flagfish has found a significant effect of condition (measured as the residuals

from a regression of log weight to log length) on variance in male color expression in

response to temperature and salinity treatments (St. Mary et al. 2001), suggesting that

environmental stresses play a role in color expression. Furthermore, Johnstone (1995)

indicates that color expression is typically influenced by both measures of nutritional

status and parasite load. Parasites have been found to impede the uptake of carotenoids









from the gut, limiting the development of color expression in fish (Milinski & Bakker

1990; Houde & Torrio 1992). This measure would be of interest to investigate in future

research on male color expression in flagfish.

Measures of parental care and courtship behavior were also expected to show

condition-dependent expression. Courtship has often been shown to be expensive

(Vehrencamp et al. 1989; Hoglund et al. 1992) requiring considerable energy reserves to

be maintained over time. Thus, a positive relationship was expected between condition

and the frequency and/or time spent performing energetically expensive behaviors such

as fanning and chasing. No such relationship was found with regard to chasing. However,

counter to our expectations fanning was negatively correlated with fat reserves. This loss

in condition could be due to energy expenditure lost through displaying. Alternatively,

this may suggest that males in poor condition increased their signaling effort at the

expense of future mating as has been demonstrated in sticklebacks (Candolin 1999).

Furthermore, males in low condition may be able to adjust the amount of energy

they invest into courtship depending on the social context. For example, Candolin (2000)

demonstrated that male sticklebacks increased red coloration in the presence of females

and the absence of males when the cost of cheating was low in terms of harassment by

dominant males. Male condition may play a more important role in flagfish under

different social contexts as male-male competition may ensure honest signaling as also

suggested in sand gobies (Svensson & Forsgren 2003).

Females were expected to allocate reproductive resources in response to diet -

dependent differences among males. However, high and low food males were not found

to differ significantly in many of the traits measured. Regardless, females were expected









to use variation in morphological characters among males in egg allocation decisions. No

such patterns were found. The lack of significant results regarding female egg allocation

could be explained by the occurrence of filial cannibalism as male flagfish are known to

consume their eggs (Klug & St. Mary 2005). However, spawning mats were checked at a

minimum of four times a day to prevent filial cannibalism from influencing egg counts.

In fish, egg size has been found to be positively associated with larval size and

hence offspring survival (Ware 1975; Marsh 1986; Pepin 1991; Einum & Fleming 1999).

This pattern is also supported from results of the current study where larval length

increased with egg area. Thus egg size may be one valuable way in which females benefit

from increased investment. Counter to our expectations, males of a lower weight for a

given length fathered larger larvae. Although, the current study found no significant

effect of male condition on egg size, a negative pattern was apparent. These results

suggest that females may allocate larger eggs with more resources to males in poor

condition.

Granted, male condition had no influence on egg energy, differential investment

can not be excluded as resource variation may exist that was not detectable by the general

methods used in the current study. For example, the direct transfer of proteins

(hormones), immune factors, and mRNA to eggs and larvae has been demonstrated in

oviparous fishes (Heath & Blouw 1998) and in some cases has been correlated with

increased larval size and survival (Ayson & Lam 1993). Yolk may be an additional

resource to measure directly in future research as initial yolk volume was found to be

correlated with larval length at hatching in capelin, Mallotus villosus (Chambers et al.

1989). Furthermore, Kolm & Olsson (2003) propose that female Banggai cardinalfish,









Pterapogon kauderni, re-direct resources into eggs extremely close to spawning,

suggesting that females may have more control over egg investment than previously

implied.

Initially, one may interpret these unexpected results as the female's adaptive

response to low quality males. Females mating with unattractive males have been found

to increase maternal investment to enhance offspring health as a result of a decrease in

viability associated with low quality fathers (Bluhm & Gowaty 2004). For example, in

collared flycatchers, females have been found to compensate for low quality parental care

of young sires by increasing yolk testosterone concentrations which boosts begging

activity (Michl et al. 2004) and in barn swallows, females manipulated egg carotenoid

levels in response to unattractive males whose offspring may have greater exposure to

parasites (Saino et al. 2002b).

However in the current study, males with low fat reserves spent more time fanning

their nests during courtship, suggesting that females allocate larger eggs and hence larger

larvae to those males who are likely to provide better quality parental care and hence

possibly greater survivorship of offspring. The proportion of time spent fanning the nest

during courtship and when eggs were received was highly correlated, suggesting that pre-

spawning behavior indicates the quality of parental care a female may expect post-

spawning. Alternatively, paternal effects can not be ruled out as a source of these

unexpected results. Genetic differences among males may have influenced larval length

as has been demonstrated in other studies which contribute offspring size and growth

rates to paternal effects (Reynolds & Gross 1992).









Consistent with the "good parent process" of sexual selection which argues that

parental care functions as a cue in mate choice (Hoelzer 1989), female flagfish also

responded to variation in fanning during courtship in addition to nest defense when

selecting mates. The direct benefits associated with parental care behaviors have been

investigated in the flagfish as nest defense has been shown to increase egg survivorship

(Klug et al. 2005) and fanning is hypothesized to reduce fungal infection (St. Mary et al.

2001; 2004) in addition to aiding in the replacement of oxygen. Hence female flagfish

select mates that provide better parental care and offer increased offspring survivorship as

has been demonstrated in other fish species (Forsgren 1997; Ostlund, & Ahnesjo 1998)

In summary, the DAH suggests that the attractiveness of a mate influences the

value of the breeding attempt. Females should allocate more resources when mating with

an attractive male as the potential for direct and/or indirect benefits received from

reproduction are greater. Measures of attractiveness such as body size, condition, fat

reserves, and color expression did not influence female mating and egg allocation

decisions. However, male condition had a significant affect on larval length, suggesting

that females produce eggs that produce larger larvae when mating with males in lower

condition. Counter to our expectations, males with low energy reserves fanned more pre

and post spawning suggesting that females respond to variation in parental care by

investing more in reproduction.

DAH has important implications for the field of sexual selection and should be

studied further as it potentially poses a problem for studies that support "good genes"

models of sexual selection. If maternal effects have not been accounted for, evidence for

increased survival of offspring fathered by attractive males may be attributable to









differential female investment rather than paternal genetic effects. Evidence for

differential allocation will improve our understanding of all factors which affect offspring

fitness. Furthermore, if conditions offspring experience during development influences

the expression of sexually selected characters in adulthood, differential allocation could

have a strong influence on the direction of sexual selected traits within a given species

(Qvarnstrom & Price 2001).

Despite support for differential allocation in a variety of taxa, experimental tests

of the hypothesis in fish is lacking besides those of Kolm 2001, Kolm & Olsson 2003 and

the current study. Given the prevalence of male parental care in fishes (Gross & Sargent

1985) and the importance of perceived benefits in differential allocation theory, future

research should aim to expand the number offish species in which this topic is

investigated.
















LIST OF REFERENCES


Andersson, M. 1994. Sexual Selection. Princeton, New Jersey: Princeton University
Press.

Ayson, F.G., & Lam, T.J. 1993. Thyroxine injections of female rabbitfish (Siganus
guttatus) broodstock; Changes in thyroid hormonal levels in plasma, eggs, and
yolk-sac larvae, and its effect on larval growth and survival. Aquaculture, 109, 83-
93.

Bluhm, C.K., & Gowaty, P. 2004. Reproductive compensation for offspring viability
deficits by female mallards, Anasplatyrhynchos. Animal Behaviour, 68, 985-992.

Burley, N. 1986. Sexual selection for aesthetic traits in species with biparental care.
American Naturalist, 127, 415-445.

Burley, N. 1988. The differential-allocation hypothesis: An experimental test. American
Naturalist, 132, 611-628.

Bolger, T., & Connolly, P.L. 1989. The selection of suitable indices for the measurement
and analysis of fish condition. Journal ofFish Biology, 26, 171-182.

Boschung, H.T. Jr., Williams, J.D., Gotshall, D.W., Caldwell, M.C., & Caldwell, D.K.
2000. The Audubon Society Field Guide to North American Fishes, Whales and
Dolphins. Chanticleer, New York.

Candolin, U. 1999. The relationship between signal quality and physical condition: is
sexual signalling honest in the three-spined stickleback? Animal Behaviour, 58,
1261-1267.

Candolin, U. 2000. Male-male competition ensures honest signaling of male parental
ability in the three spined stickleback (Gasterosteus aculeatus). Behavioral
Ecology and Sociobiology, 49, 57-61.

Campanella, P.J., & Wolf, L.L. 1974. Temporal leks as a mating system in a temperate
zone dragonfly (Odonata: Anisoptera). I. Plathemis lydia (Drury). Behaviour, 51,
49-87.

Chambers, R.C., Leggett, W.C., & Brown, J.A. 1989. Egg size, female effects, and the
correlations between early life-history traits of capelin, Mallotus villosus: An
appraisal at the individual level. Fish Bulletin, 87, 515-523.









Cunningham, E.J.A., & Russell, A.F. 2000. Egg investment is influenced by male
attractiveness in the mallard. Nature, 404, 74-76.

Darwin, C. 1871. The Descent ofMan and Selection in Relation to Sex. Murray, London.

Einum, S., & Fleming, I.A. 1999. Maternal effects of egg size in brown trout (Salmo
trutta): Norms of reaction to environmental quality. Proceedings of the Royal
Society of London Series B, 266, 2095-2100.

Edvardsson, M., & Arnqvist, G. 2005. The effects of copulatory courtship on differential
allocation in the red flour beetle, Tribolium castaneum. Journal ofInsect Behavior,
18, 313-322.

Folstad, I., & Karter, A. J. 1992. Parasites, bright males, and the immunocompetence
handicap. American Naturalist, 139, 603-622.

Forsgren, E. 1997. Female sand gobies prefer good fathers over dominant males.
Proceedings of the Royal Society of London Series B, 264, 1283-1286.

Forsgren, E., Kvarnemo, C., & Lindstrom, K. 1996. Mode of sexual selection determined
by resource abundance in two sand goby populations. Evolution, 50, 646-654.

Foster, N.R., Cairns, J. Jr., & Kaesler, R.L. 1969. The flagfish, Jordanellafloridae, as a
laboratory animal for behavioral bioassay studies. Proceedings of the Academy of
Natural Sciences Philadelphia, 121, 129-152.

Gil, D., Graves, J., Hazon, N., & Wells, A. 1999. Male attractiveness and differential
testosterone investment in zebra finch eggs. Science, 286, 126-128.

Grafen, A. 1990. Biological signals as handicaps. Journal of Theoretical Biology, 144,
517-546.

Grether, G.F., Hudon, J., & Endler, J.A. 2001. Carotenoid scarcity, synthetic pteridine
pigments and the evolution of sexual coloration in guppies (Poecilia reticulata).
Proceedings of the Royal Society ofLondon, Series B, 268, 1245-1253.

Gross, M.R., & Sargent, R.C. 1985. The evolution of male and female parental care in
fishes. American Zoologist, 25, 807-822.

Gwinner, H. & Schwabl, H. 2005. Evidence for sexy sons in European starlings (Sturnus
vulgaris). Behavioral Ecology and Sociobiology, 58, 375-382.

Hamilton,W.D., & Zuk, M. 1982. Heritable true fitness and bright birds: A role for
parasites? Science, 218, 384-386.

Heath, D.D., & Blouw, D.M. 1998. Are maternal effects in fish adaptive or merely
physiological side effects? In Maternal Effects as Adaptations (Mousseau, T.A., &
Fox, C.W. eds) pp 178-201. New York: Oxford University Press.






50


Hoelzer, G.A. 1989. The good parent process of sexual selection. AnimalBehaviour, 38,
1067-1078.

Hoglund, J., Kdlds, J.A., & Fiske, P. 1992. The costs of secondary sexual characters in
the lekking great snipe. (Gallinago media). Behavioral Ecology and Sociobiology,
30, 309-315.

Holm, C.H. 1973. Breeding sex ratios, territoriality, and reproductive success in the red-
winged blackbird (Agelaiusphoeniceus). Ecology, 54, 356-365.

Houde, A.E., & Torio, A.J. 1992. Effect of parasitic infection on male colour pattern and
female choice in guppies. Behavioral Ecology, 3, 346-351.

Hurst, D.T. 1980. An Introduction to the Chemistry and Biochemistry ofPyrimidines,
Purines and Pteridines. New York: John Wiley.

Johnstone, R.A. 1995. Sexual selection: Honest advertisement and the handicap principle:
Reviewing the evidence. Biological Reviews, 70, 1- 65.

Kangas, N., & Lindstrom, K. 2001. Male interactions and female mate choice in the sand
goby, Pomatoschistus minutus. Animal Behaviour, 61, 425-430.

Kempenaers, B., Verheyen, G.R., & Dhondt, A.A. 1997. Extra-pair paternity in the blue
tit (Parus caeruleus): Female choice, male characteristics, and offspring quality.
Behavioral Ecology, 8, 481-492.

Klug, H., & St.Mary, C.M. 2005. Reproductive fitness consequences of filial cannibalism
in the flagfish, Jordanellafloridae. Animal Behaviour, 70, 685-691.

Klug, H., Chin, A., & St. Mary, C.M. 2005. The net effects of guarding on egg
survivorship in the flagfish, Jordanellafloridae. Animal Behaviour, 69, 661-668.

Knapp, R.A. 1995. Influence of energy reserves on the expression of a secondary sexual
trait in male bicolor damselfish, Stegastes partitus. Bulletin ofMarine Science, 57,
672-681.

Knapp, R.A., & Kovach, J.T. 1991. Courtship as an honest indicator of male parental
quality in the bicolor damselfish, Stegastespartitus. Behavioral Ecology, 2, 295-
300.

Kodric-Brown, A. 1989. Dietary carotenoids and male mating success in the guppy; and
environmental component to female choice. Behavioral Ecology and Sociobiology
25, 393-401.

Kodric-Brown, A. 1996. Role of male-male competition and female choice in the
development of breeding coloration in pupfish (Cyprinodonpecosensis).
Behavioral Ecology, 7, 431-437.









Kodric-Brown, A. 1998. Sexual dichromatism and temporary color changes in the
reproduction of fishes. American Zoology, 38, 70-81.

Kodric-Brown, A., & Brown, J.J. 1984. Truth in advertising: The kinds of traits favored
by sexual selection. American Naturalist, 124, 309-323.

Kodric-Brown, A. & Nicoletto, P.F. 1993. The relationship between physical condition
and social status in pupfish, Cyprinodonpecosensis. Animal Behaviour, 46, 1234-
1236.

Kolm, N. 2001. Females produce larger eggs for large males in a paternal mouth
brooding fish. Proceedings of the Royal Society of London Series B, 268, 2229-
2234.

Kolm, N., & Olsson, J. 2003. Differential investment in the Banggai cardinalfish: Can
females adjust egg size close to egg maturation to match the attractiveness of a new
partner? Journal ofFish Biology, 63, 144-151.

Kvarnemo, C., Forsgren, E., & Magnhagen, C. 1995. Effects of sex ratio on intra- and
inter sexual behavior in sand gobies. Animal Behaviour, 50, 1455-1461.

Lindstrom, K. 1998. Energetic constraints on mating performance in the sand goby.
Behavioral Ecology, 9, 297-300.

Marsh, E. 1986. Effects of egg size on offspring fitness and maternal fecundity in the
orange-throat darter, Eihevi,\ t,uai spectabile (Pisces: Percidae). Copeia, 18-30.

Mertz, J.C., & Barlow, G.W. 1966. On the reproductive behavior of Jordanellafloridae
(Pisces: Cyprinodontidae) with special reference to a quantitative analysis of
parental fanning. Zeitschrift fir Tierpsychologie. 23, 537-554.

McEdward, L.R., & Coulter, L.K. 1987. Egg volume and energetic content are not
correlated among sibling offspring of starfish: Implications for life-history theory.
Evolution, 41, 914-917.

Michl, G., Toirk, J., Peczely, P., Garamszegi, L.Z., & Schwabl, H. 2004. Female collared
flycatchers adjust yolk testosterone to male age, but not to attractiveness.
Behavioral Ecology, 16, 383-388.

Milinski, M., & Bakker, C.M. 1990. Female sticklebacks use male coloration in mate
choice and hence avoid parasitized males. Nature, 344, 330-333.

Moller, A.P. 1990. Effects of a haematophagous mite on the barn swallow (Hirundo
Rustica): A test of the Hamilton and Zuk hypothesis. Evolution, 44, 771-784.

Nicoletto, P.F. 1993. Female sexual response to condition-dependent ornaments in the
guppy, Poecilia reticulata. Animal Behaviour, 46, 441-450.






52


Nisbet, I.C.T. 1973. Courtship-feeding, egg-size, and breeding success in common terns.
Nature, 241, 141-142.

Norris, K. 1990. Female choice and the quality of parental care in the great tit, Parus
major. Behavioral Ecology and Sociobiology, 27, 275-281.

Norris, K. 1993. Heritable variation in a plumage indicator of viability in male great tits,
Parus major. Nature, 362, 537-539.

Olson, V.A., & Owens, I.P.F. 1998. Costly sexual signals: Are carotenoids rare, risky or
required? Trends in Evolution and Ecology, 13, 510-514.

Ostlund, S., & Ahnesjo, I. 1998. Female fifteen-spined sticklebacks prefer better fathers.
Animal Behaviour, 56, 1177-1183.

Page, L. M., & Burr, B.M. 1991. A Field Guide to Freshwater Fishes: North America
North of Mexico. New York: Houghton Mifflin Company.

Parker, T.H. 2003. Genetic benefits of mate choice separated from differential maternal
investment in red junglefowl (Gallus gallus). Evolution, 57, 2157-2165.

Petrie, M. 1983. Female moorhens compete for small fat males. Science, 220, 413- 415.

Petrie, M. 1994. Improved growth and survival of offspring of peacocks with more
elaborate trains. Nature, 371, 598-599.

Petrie, M., & Williams, A. 1993. Peahens lay more eggs for peacocks with larger trains.
Proceedings of the Royal Society of London Series B, 251, 127-131.

Pepin, P. 1991. Effects of temperature and size on development, mortality, and survival
rates of the pelagic early life-history of marine fish. Canadian Journal ofFisheries
andAquatic Science, 48, 503-518.

Qvarnstrom, A., & Price, T.D. 2001. Maternal effects, paternal effects and sexual
selection. Trends in Ecology and Evolution, 16, 95-100.

Reyer, H.U., Frei, G., & Som, C. 1999. Cryptic female choice: frogs reduce clutch size
when amplexed by undesired males. Proceedings of the Royal Society of London
Series B, 266, 2101-2107.

Reynolds, J.D., & Gross, M.R. 1992. Female mate preference enhances offspring growth
and reproduction in a fish, Poecilia reticulata. Proceedings of the Royal Society of
London Series B, 250, 57-62.

Ridgway, M.S., & Shuter, B.J. 1994. The effects of supplemental food on reproduction in
parental male smallmouth bass. Environmental Biology ofFishes, 39, 201-207.









Saino, N., Paola Ferrari, R., Martinelli, R., Romano, M., Rubolini, D., & Moller, A.P.
2002a. Early maternal effects mediated by immunity depend on sexual
ornamentation of the male partner. Proceedings of the Royal Society of London
Series B, 269, 1005-1009.

Saino, N., Bertacche, V., Paola Ferrari, R., Martinelli, R., Pape Moller, A., & Stradi, R.
2002b. Carotenoid concentration in barn swallow eggs is influenced by laying
order, maternal infection and paternal ornamentation. Proceedings of the Royal
Society of London Series B, 269, 1729-1733.

Sheldon, B.C., 2000. Differential allocation; Tests, mechanisms, and implications. Trends
in Ecology and Evolution, 15, 397-401.

Sheldon, B.C., Andersson, S., Griffith, S.C., Ornborg, J., & Sendecka, J. 1999.
Ultraviolet colour variation influences blue tit sex ratios. Nature, 874-877.

Smith, W.E. 1973. A Cypriodontid fish, Jordanellafloridae, as a laboratory animal for
rapid chronic bioassays. Journal of the Fisheries Research Board of Canada, 30,
329-330.

Smith, C., & Wootton, R. J. 1999. Parental energy expenditure of the male three-spined
stickleback. Journal ofFish Biology, 54, 1132-1136.

St. Mary, C.M., Noureddine, C.G., & Lindstrom, K. 2001. Environmental effects on male
reproductive success and parental care in the Florida flagfish Jordanellafloridae.
Ethology, 107, 1035-1052.

St. Mary, C.M., Gordon, E., & Hale, R.E. 2004. Environmental effects on egg
development and hatching success in Jordanellafloridae, a species with parental
care. Journal ofFish Biology, 65, 1-9.

Svensson, O., & Forsgren, E. 2003. Male mating success in relation to food availability in
the common goby. Journal ofFish Biology, 62, 1217-1221.

Szentirmai, I., Komdeur, J., & Szekely, T. 2005. What makes a nest-building male
successful? Male behavior and female care in penduline tits. Behavioral Ecology,
994-1000.

Thomhill, R. 1976. Sexual selection and nuptial feeding behaviour in Bittacus apicalis
(Insecta, Mecoptera). American Naturalist, 110, 529-548.

Vehrencamp, S.L., Bradbury, J.W., & Gibson, R.M. 1989. The energetic cost of display
in male sage grouse. Animal Behaviour, 38, 885-896.

Ware, D.M. 1975. Relation between egg size, growth, and natural mortality of larval fish.
Journal of the Fisheries Research Board of Canada, 32, 33-41.






54


Wiggins, D.A., & Morris, R.D. 1986. Criteria for female choice of mates: Courtship
feeding and parental care in the common tern. American Naturalist, 128, 126-129.

Zahavi, A. 1975. Mate selection- a selection for a handicap. Journal of Theoretical
Biology, 53, 205-214.















BIOGRAPHICAL SKETCH

In June of 2003 I received a Bachelor of Science degree from Ohio State

University. As an undergraduate I was able to focus on three major research projects in

various disciplines of zoology. These projects occurred in the areas of neuroethology,

pharmacology, aquatic ecology, and mating systems.

I began at the Rothenbuhler Honey Bee Lab, assisting in research aimed at

understanding the role of nitric oxide in the feeding behavior of the adult sphinx moth,

Manduca sexta. In April of 2002, I presented this research at the 24th Annual Meeting for

the Association of Chemoreception Sciences in Sarasota, Florida. In May of 2002, I

continued to present this research at several undergraduate research symposiums. As a

result, I received the Arts and Sciences Award for Excellence in Scholarship and was

given the opportunity to travel abroad in November of 2002 to attend the University of

Sao Paolo's Symposium of Undergraduate Research in Brazil.

Following this experience, I worked as a graduate student assistant at the Aquatic

Ecology Lab. I gained invaluable field experience working in Ohio's reservoirs, aiding

in research that sought to determine the impact of stocked Saugeye on the abundance of

their primary prey, the Gizzard Shad. I became familiar with techniques used to collect

larval and adult fish, in addition to methods used for ageing fish.

Throughout my undergraduate years I volunteered in the lab of Jerry Downhower.

This research focused on the costs of compromised female choice and the consequences

of estrogen-like compounds on mating behavior in Japanese killifish, Oryzias latipes. It









was from this experience that I became interested in the study of sexual selection. I

decided to pursue my interests further by attending graduate school at the University of

Florida and studying the mating behavior of flagfish, Jordanellafloridae, in the lab of

Colette St. Mary.

As an undergraduate I was grateful to be given the opportunity to participate as a

research assistant in several laboratories. As a graduate student with my own research

program I have extended this opportunity to students at the University of Florida by

mentoring two undergraduates. Each student aided in the collection of data. Furthermore,

over the past eight semesters I have also interacted with the student body by teaching

laboratory courses in introductory biology, evolution and ecology. I now hope to pursue

a career in science education.