Salinity-related fitness differences amongst GPI genotypes in the mosquitofish Gambusia holbrooki (Poeciliidae: Teleostei)

Salinity-induced mortality was examined in the mosquitofish Gambusia holbrooki to determine if natural selection associated with the tidal inundation of lower reach populations could potentially maintain glucose-6-phosphate isomerase ( GPI -2; E.C. 5.3.1.9) allele frequency variation observed at field sites. In vitro experiments established if survival was independent of GPI -2 genotype, and/or standard body length (S.L.). Bioassay results at 17%₀ were consistent with lower survival of GPI -2*88/88 homozygote genotypes, GPI -2*100/100 genotypes in juvenile size classes, and greater survival of GPI -2*109/100 genotypes; 100% mortality was observed at salinities of 30%₀. In the early stages of the 30%₀ bioassay, juvenile fish carrying each GPI -2 allele suffered similar, high rates of mortality. Following 19 weeks exposure to 4–8%₀ NaCl the frequency of GPI -2*88 alleles decreased in adult G. holbrooki (> 17 mm SL). Salinity-induced mortality was found to be a selective influence capable of creating and maintaining GPI -2 allele frequency differences among G. holbrooki subpopulations, and/or between sex/size cohorts within subpopulations, but only at intermediate salinities. Whether saltwater inundation at field sites maintains inter-population diversity and produces local adaptation will be determined by the age structure of the population during periods of exposure, fluctuations in salinity as determined by the relationship between rainfall and tidal regime, and the level of immigration from non-affected sites.     

Sexual selection for small size in male mosquitofish (Gambusia holbrooki)

In many poeciliid fishes, large males which court females coexist with small males which sneak-copulate. It is unclear whether these two tactics represent two evolutionarily stable strategies or if sneaking is a conditional strategy adopted by small, unattractive males. We studied the success of sneaky copulation by looking for sperm in the gonoduct of females after they were kept for 48 h with a male. A logistic regression analysis showed that the probability of a female being inseminated increased with female length and decreased with male length. The length of the male relative to that of the female was the best predictor of success. This result was confirmed using virgin females, thereby excluding any possible confounding effect due to the release of sperm from previous copulations. Sperm counts suggested that large males do not compensate for their reduced copulatory success by releasing larger sperm numbers. Behavioural data indicate that the advantages to small males are twofold: they have a greater chance to approach females from behind without being detected, and manoeuvre better when inserting the gonopodium into the female''s gonoduct. The selective advantage of small size might explain male dwarfism in poeciliids. Our results also suggest that small males adopting the sneaky tactic may be as successful as large males adopting courtship, and that alternative mating strategies may be maintained by negative density-dependent selection.     

Behavioral constraints for the spread of the eastern mosquitofish, Gambusia holbrooki (Poeciliidae)

Eastern mosquitofish (Gambusia holbrooki) are native to the southeastern United States but notoriously invasive elsewhere, and are aggressive predators in ecosystems they inhabit. Information on dispersal behavior is needed to better understand mosquitofish spread upon introduction and potential means to mitigate that spread. We experimentally tested the effects of shallow water depths (3–24 mm) and obstacles (leaf litter) on mosquitofish dispersal behavior, plus a range of conditions relevant to field situations. Mosquitofish dispersed significantly faster in deeper water (p < 0.001) but some dispersed in only 3 mm water depth (i.e., one-half average body depth). Wetland and upland leaf litter at natural densities strongly interfered with mosquitofish dispersal behavior. Based on our results, introduced mosquitofish spread rapidly given unimpeded dispersal corridors (e.g., mowed ditches), and may do so at rates >800 m/day. Also, consistent lack of sexual dimorphism in dispersal behavior indicates that mosquitofish spread is not strongly dependent on female poeciliid reproductive biology. Our results support designation of mosquitofish as highly invasive and suggest that barriers to mosquitofish spread must obstruct dispersal pathways as shallow as 3 mm depth.     

Predation and the persistence of melanic male mosquitofish (Gambusia holbrooki)

The empirical reasons for the persistent rarity of a genotype are typically complex and tedious to identify, particularly in nature. Yet rare morphs occur in a substantial fraction of phenotypic polymorphisms. A colour polymorphism has persisted for decades in the eastern mosquitofish, yet why this is so remains obscure. Here, I report the results of (1) intensive sampling at 45 natural sites to obtain the frequency distribution of the melanic (black) mosquitofish morph in Florida, (2) predation trials, conducted independently in mesocosms, with three different predatory species and (3) two mark-recapture studies, conducted in nature. This work (1) documents the rarity of melanic mosquitofish in nature, (2) demonstrates that melanic males experience a selective advantage over silver males in the presence of predators, (3) indicates no difference in the colour morphs' survival at a natural site essentially devoid of these predators, although suggesting a higher rate of recapture for melanic males at a site rife with predators. Overall, selective predation appears to contribute to the persistence of the melanic morph, despite its rarity in nature.     

Life history of Gambusia holbrooki (Pisces, Poeciliidae) in the Ebro delta (NE Iberian peninsula)

We studied age, growth and reproduction of an introduced poeciliid, Gambusia holbrooki (Agassiz, 1859), in the Ebro delta marshes. Maximum ages observed were 2⁺ in females and 0⁺ in males. Growth commenced in March and continued until September. Females reached greater sizes, had a higher rate of growth and lived longer than males. The largest sizes found were 63 mm versus 32 mm . Multiple spawning occurred throughout the reproductive cycle (from May to September). The average brood size per female reached its maximum in July in the parental cohort (86 embryos/) and in August in the offspring cohort (29 embryos/). After reproduction the parental cohort decreased drastically. The relationship between fecundity (F) and total length (mm) was calculated for each month of the reproductive period. Sex-ratio deviated significantly from unity.Viviparity, several broods and large larvae provide G. holbrooki with several advantages over oviparous species. The capacity to adjust its life history to a particular environment could explain the life history differences between those in the Ebro delta and other populations.     

The testicular cycle of Gambusia affinis holbrooki (Teleostei: Poeciliidae)

Fifteen male mosquito fish ( Gambusia affinis holbrooki ) were collected in 1989 on the 15th of each month to perform a quantitative histologic study of the annual testicular cycle including a calculation of the gonadosomatic index, testicular volume, and the total volume per testis occupied by each germ cell type. The cycle comprises two periods: spermatogenesis and quiescence. The spermatogenic period begins in April with the development of primary spermatogonia into secondary spermatogonia, spermatocytes and round spermatids. In May, the first spermatogenic wave is completed and the testicular volume begins to increase up to June when the maximum testicular volume and gonadosomatic index are reached. Germ cell proliferation with successive spermatogenetic waves continues until August. In September germ cell proliferation ceases and neither secondary spermatogonia nor spermatocytes are observed. However, spermiogenesis continues until October. In November, spermiogenesis has stopped and the testis enters the quiescent period up to April. During this period only primary spermatogonia and spermatozoa are present in the testis. In addition, a few spermatids whose spermiogenesis was arrested in November are observed. Testicular release of spermatozoa is continuous during the entire spermatogenesis period. The spermatozoa formed at the end of this period (September-October) remain in the testis during the quiescent period and are released at the beginning of the next spermatogenesis period in April. Developed Leydig cells appear all year long in the testicular interstitium, mainly around both efferent ducts and the testicular tubule sections showing S₄ spermatids.     

Characteristics of dispersal in the eastern mosquitofish Gambusia holbrooki

The demographic and genetic characteristics of dispersing Gambusia holbrooki were examined relative to changes in stream flow velocity, flow direction, and rapid saltwater inundation. Dispersal of all G. holbrooki sex classes decreased as flow velocity increased, dispersal for a given flow velocity being greater in female, than male and juvenile fish respectively. Juveniles and males dispersed at low (9 mm s⁻¹) but not high (109 mm s ¹) flow. Females dispersed at both flow rates. Dispersing adults were not larger than non-dispersing adults of the same sex. Direction of dispersal was sex and size dependent, but independent of flow velocity. Larger females (>22 mm) moved preferentially downstream, while smaller females (17–22 mm) remained stationary or moved in the upstream direction. Males dispersed equally in both directions. No relationship was found between dispersal characteristics and the presence of high salt concentrations. Movement patterns at high flow were independent of glucose-6-phosphate isomerase ( GPI-2* ) alleles carried by dispersing individuals. Significant non-random dispersal of sex or size cohorts, and the interaction of dispersal with flow velocity have significant implications for the maintenance of genetic diversity in stream ecosystems, particularly where localized selection or genetic drift may alter the genetic constitution of specific sex or size cohorts. The applicability of these results to field populations is discussed.     

Mechanisms of sexual selection operating on body size in the mosquitofish (Gambusia holbrooki)

Collections in two natural populations of the mosquitofish,Gambusia holbrooki, were made to determine whether the sexesassociated nonrandomly based on body size, and laboratory experimentswere performed to identify mechanisms that could generate theobserved field patterns of association. In both populationslarger females were associated with larger males. However, thesize distributions of males caught with and without femaleswere not different. Both gravid females and females that hadreleased broods within the previous 24 h spent more time nearlarger males in laboratory choice experiments. In contrast,males did not discriminate between females on the basis of sizeor reproductive status in laboratory choice experiments, andmales did not attempt relatively more copulations with largerfemales when one male was allowed to freely interact with twofemales. Body size differences and aggressive interactions betweenmales did influence the access of males to females, with largermales attempting more copulations with females when two maleswere allowed to freely interact with one female: aggressiveinteractions between males were frequent when males differedby < 1.0 mm standard length but were rare when males differedby>5.0 mm standard length. These results indicate that femalechoice for larger males could contribute to generating the nonrandomassociations between the sexes that were observed in these twopopulations and that interactions between males in the vicinityof females may additionally increase the mating advantages oflarger males.     

Sex differences in compensatory and catch-up growth in the mosquitofish Gambusia holbrooki

In many taxa, temporary nutritional shortage early in development can favour compensatory strategies that include elevated growth (‘compensatory growth’) and/or extension of the usual period of development (‘catch-up growth’) once conditions improve. The net gains from each strategy depend on the extent to which larger body size increases fitness relative to associated costs (e.g. long-term effects on adult performance, or a greater risk of juvenile mortality). These costs and benefits are likely to differ between the sexes due to sex-specific selection. We documented the responses of male and female mosquitofish (Gambusia holbrooki) to 3 weeks of low food availability (7–28 days old) that restricted subsequent growth and morphology compared to control fish continuously reared on a high food diet (N = 635 fish total). Neither sex elevated their growth rate immediately after being returned to a normal diet compared to control fish. When measured over the entire period until maturation, however, females showed compensatory growth. Males did not. Both sexes also exhibited catch-up growth but the delay until maturation was significantly longer for males. Despite early growth restriction, both sexes eventually matured at almost the same size as control fish, although males had a significantly smaller gonopodium (a sexually selected trait) than that of control males. Reasons for these sex differences are discussed.     

Life history and parasites of the invasive mosquitofish (Gambusia holbrooki) along a latitudinal gradient

The eastern mosquitofish (Gambusia holbrooki) is among the most invasive fish worldwide and yet, while very abundant in most Mediterranean countries, it is unable to tolerate the colder winters of northern and central Europe. Understanding the effects of latitude on its life-history traits is essential to predict the potential for its invasion of central Europe in current scenarios of climate change. We studied the variation of life-history traits and parasite load in the eastern mosquitofish along a latitudinal gradient from southern France to southern Spain, sampling mosquitofish populations in eight Mediterranean river mouths ranging 5° in latitude. Southern mosquitofish populations displayed higher catch rates, allocated more energy to reproduction (gonadosomatic index and gonadal weight after accounting for fish size) and had a lower condition (total weight and eviscerated weight after accounting for fish size) than in northern populations. Despite variability among populations, size-at-maturity (L₅₀) significantly varied with latitude and northern individuals matured at smaller size (lower L₅₀). Parasite prevalence ranged from 0.0 to 26.7% but parasite richness was very low; all the parasites identified were larvae of pleurocercoid cestodes belonging to the order Pseudophyllidea. The abundance of mosquitofish parasites decreased with latitude and the presence and number of parasites infecting the mosquitofish had a significant negative effect on fish condition. The significant effects of latitude on the catch rates, life history and parasites of mosquitofish highlight the importance of latitudinal studies of invasive species to understand the interactive mechanisms of climate change and biological invasions.     

Beta-naphthoflavone inhibits the induction of hepatic oestrogen-dependent proteins by 17alpha-ethynylestradiol in mosquitofish (Gambusia holbrooki)

The interactive effects of an aryl hydrocarbon receptor (AhR) agonist and of a xenoestrogen on biomarker responses were studied in the liver of male mosquitofish (Gambusia holbrooki). Hepatic 7-ethoxyresorufin O-deethylase (EROD) enzymatic activity was measured as a biomarker of exposure to the model AhR agonist beta-naphthoflavone (bNF). Hepatic proteins indicating the exposure of males to the synthetic oestrogen 17alpha-ethynylestradiol (EE2) were monitored by Western blot analysis using immunoserum prepared for this study. After a semi-static exposure only to waterborne EE2, Western blot analysis of liver homogenate revealed the induction of two protein bands (a double band at 205 kDa and a single band at 125 kDa). The interaction between bNF and EE2 was investigated by analysing, on the one hand, EROD activity and, on the other hand, immunoreactivity corresponding to the two oestrogen-dependent protein bands in the liver of fish exposed to different concentrations of bNF for 2 days, then to the same concentrations of bNF plus 0.1 µg l^-1 EE2 for 5 days. EE2 changed neither the basal activity of EROD nor its rate of induction with 1.0 and 4.0 µg l^-1 bNF. On the other hand, the induction of oestrogen-dependent proteins with 0.1 µg l^-1 EE2 was inhibited by exposure to 4.0 µg l^-1 bNF. These results together with literature data suggest that field monitoring of xenoestrogen contamination through the analysis of oestrogen-dependent protein in male fish as a biomarker should take into account the possible negative interference of AhR agonists.     

Melanic body colour and aggressive mating behaviour are correlated traits in male mosquitofish (Gambusia holbrooki)

Correlated traits are important from an evolutionary perspective as natural selection acting on one trait may indirectly affect other traits. Further, the response to selection can be constrained or hastened as a result of correlations. Because mating behaviour and body colour can dramatically affect fitness, a correlation between them can have important fitness ramifications. In this work, melanic (black) male mosquitofishes (Gambusia holbrooki) with temperature-sensitive body-colour expression are bred in captivity. Half of the sons of each melanic sire are reared at 19 degrees C (and express a black body colour) and half are reared at 31 degrees C (and express a silver body colour). The two colour morphs are placed in the same social setting and monitored for behavioural differences. Mating behaviour and colour are correlated traits. Mating behaviour differs markedly between the two phenotypes, despite high genetic relatedness. Melanic (black) phenotypes are more aggressive towards females, chasing them and attempting more matings than their silver siblings. Females avoid melanic-male mating attempts more than silver-male mating attempts. When males with temperature-sensitive colour expression are melanic and aggressive, they probably experience a very different selective regime in nature from when they are silver and less aggressive. Under some conditions (e.g. predation), melanic coloration and/or aggression is advantageous compared with silver coloration and/or less aggressive behaviour. However, under different conditions (e.g. high-frequency melanism), melanism and/or aggression appears to be disadvantageous and melanic males have reduced survival and reproduction. Selective advantages to each morph under different conditions may enable the long-term persistence of this temperature-sensitive genotype.     

Consistency of leadership in shoals of mosquitofish (Gambusia holbrooki) in novel and in familiar environments

In social animal groups, an individual's spatial position is a major determinant of both predation risk and foraging rewards. Additionally, the occupation of positions in the front of moving groups is generally assumed to correlate with the initiation of group movements. However, whether some individuals are predisposed to consistently occupy certain positions and, in some instances, to consistently lead groups over time is as yet unresolved in many species. Using the mosquitofish (Gambusia holbrooki), we examined the consistency of individuals' spatial positions within a moving group over successive trials. We found that certain individuals consistently occupied front positions in moving groups and also that it was typically these individuals that initiated group decisions. The number of individuals involved in leading the group varied according to the amount of information held by group members, with a greater number of changes in leadership in a novel compared to a relatively familiar environment. Finally, our results show that the occupation of lead positions in moving groups was not explained by characteristics such as dominance, size or sex, suggesting that certain individuals are predisposed to leadership roles. This suggests that being a leader or a follower may to some extent be an intrinsic property of the individual.     

Beta-naphthoflavone inhibits the induction of hepatic oestrogen-dependent proteins by 17alpha-ethynylestradiol in mosquitofish (Gambusia holbrooki)

Quantification of metallothioneins (MTs) is classically associated with a cellular response to heavy metal contamination and is used in the monitoring of disturbed ecosystems. Despite the characterization of several MT genes in marine bivalves, only a few genetic studies have used MT genes as potential biomarkers of pollution. The aim of this study was to assess whether MT gene polymorphism could be used to monitor exposure of the Pacific oyster Crassostrea gigas to heavy metals and to develop specific genetic markers for population genetic studies in relation to environmental stress. The polymorphism of two exons of the C. gigas MT gene CgMT1 were studied using polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) in both field populations exposed to various metals concentrations and in experimentally exposed populations. High frequencies of two SSCP types in exons 2 and 3 of the CgMT1 gene have found to be significantly associated with tolerance to metals in experimental and field oyster populations. The use of MT1 gene polymorphism in C. gigas as in the present study should therefore be of high ecological relevance. In conclusion, the analysis of the types in these two CgMT1 gene exons, which can confer a greater tolerance to heavy metals, can constitute a good biomarker of effect of the presence of heavy metals in ecosystems.     

Reproductive biology of Gambusia affinis holbrooki Baird and Girard, Xiphophorus helleri (Gunther) and X. maculatus (Heckel) (Pisces; Poeciliidae) in Queensland, Australia

The reproductive biology of three introduced Poeciliidae, Gambusia affinis holbrooki Baird and Girard, Xiphophorus helleri (Günther) and X. maculatus (Heckel) was studied over a 14-month period in Brisbane, Queensland. Photoperiod and water temperature determined the length of the breeding season in G. affinis , which produced up to nine broods between August and March, with the peak of reproductive activity in October. Fecundity was linearly related to body weight in G. affinis (mean fecundity = 22.78 ± 1.02, range = 3–108 offspring per brood).
Xiphophorus helleri bred from August to May, producing eight to nine broods, with peak reproductive activity from October to December. Fecundity was curvilinearly related to body weight (mean fecundity = 60.15 ± 3.8, range = 7–183 offspring per brood. Xiphophorus maculatus bred between September and March, producing five to six broods during the study period. Fecundity was linearly related to standard length (s.l.) (mean fecundity = 27.27 ± 2.4, range = 3–92 offspring per brood). Although the reproductive strategy of the three species differs, all have the capacity to build up large populations in south-eastern Queensland. Their present distributions and relative abundance are discussed.     

Sexual selection,reproductive behavior,and speciation in the mbuna species flock of Lake Malawi (Pisces: Cichlidae)

Synopsis The cichlid species flocks of the African Great Lakes represent the most extreme case of adaptive radiation among vertebrates. Recently, attention has focused on the potential for sexual selection to drive or accelerate speciation in these fishes. Cichlids as a whole are social in nature and display complex behavior, particularly during courtship and spawning; however, the extent to which changes in species recognition cues may account for species diversity among haplochromine lineages has remained speculative. Our investigations have indicated that oral incubating haplochromines show a reduction in diversity and extent of courtship relative to substrate brooding cichlids, and apparently retain aspects of a primitive specific mate recognition system. Laboratory observations of courtship in the Malawian endemicPseudotropheus zebra suggest that organization of the spawning bout is loose, and lacking in any well defined stimulus-response chain. Interspecific comparisons of in situ courtship behavior among male mbuna, lithophilous haplochromines of Lake Malawi, revealed only one potential example of species-specific behavior, and indicated that mate choice occurred prior to the onset of intense courtship. Courtship display by male mbuna does not appear critical to species recognition and may represent an evolutionary relict. Alternatively, male courtship display may have an indirect effect on successful reproduction or may be maintained through pleiotropic effects. The mbuna display no evidence of behavioral innovation and show limited interspecific differentiation in behavioral expression. More generally, there is no evidence at present to suggest that epigamic sexual selection, acting on courtship behavior, has been a major mechanism in the diversification of the haplochromine species flocks.     

Fat cycling in the mosquitofish (Gambusia affinis): fat storage as a reproductive adaptation

Summary We argue, based on reviewed literature covering reptiles, amphibians, birds, and fish, that fat storage may represent a life history adaptation because it enables an organism to shift in time when resources are allocated to reproduction. We applied these arguments to fat and population cycles in three populations of the mosquito fish, Gambusia affinis. For males, there appeared to be a constant size at maturation during the reproductive season. Mature males became scarce late in the summer. At the same time, immature males delayed maturity and attained much larger sizes; they matured in large numbers in the fall. The amount of stored fat tended to be equal for immature and mature males at all times except in the late summer. In the August samples, when mature males were relatively rare, they also had the lowest level of fat reserves. It appears that the older generation of mature males did not store fat and did not overwinter. At the same time, immature males registered a two to three fold increase in fat reserves. These differences in fat content between mature and immature males disappeared by September, probably because of the recruitment of a new generation of mature males. The reserves were gradually utilized during the winter. Females reproduced from the late spring through mid- to late-summer. They stopped reproducing in the late summer, when there was ample time to produce an additional litter of young. There was an inverse relationship between resources devoted to reproduction and fat reserves. As reproductive allotment decreased in the late summer, fat reserves increased. The magnitude of the change in fat reserves was similar to that displayed by males. The reserves were depleted over the winter. Significant reserves remained at the beginning of the reproductive season the following spring. Reproducing females utilized the remaining reserves significantly more rapidly than non-reproducing females. An analysis of resource availability revealed an overall decrease in food availability in the late summer, coincident with the increase in fat reserves. These cycles are therefore not attributable to changes in resource availability. They instead indicate a change in how resources are allocated by the fish. The trends in the data indicate that fat reserves are used to shift investment in reproduction from the late summer to the following spring. In males, deferring maturity, rather than maturing in August, allows them to store the necessary reserves to survive the winter so that they can mate the following spring. In females, a subset of the fat reserves is intended for producing the first clutch of eggs the following spring. The female pattern corresponds to those reported for a diversity of organisms. The possible advantages of shifting reproductive effort from the fall to the following spring include higher fecundity and higher offspring fitness. The limitations of the methodology and potential directions for future research are discussed.     

Characterization and use of the total head soluble cholinesterases from mosquitofish (Gambusia holbrooki) for screening of anticholinesterase activity

Inhibition of cholinesterases (ChE) has been widely used as an environmental biomarker of exposure to organophosphates (OP) and carbamate (CB) pesticides. Different ChE isoforms may be present in the same tissue and may present distinct sensitivities towards environmental contaminants. The present work characterises the soluble ChE present in mosquitofish (Gambusia holbrooki) total head homogenates, through the use of different substrates and selective inhibitors of cholinesterasic activity. Furthermore, the effects of sodium dodecylsulphate (SDS) on the enzymatic activity were investigated, both in vivo and in vitro. These results showed that acetylcholinesterase (AChE) seemed to be the predominant form present in head homogenates of G. holbrooki, despite the inhibition by tetraisopropylpyrophosphoramide (iso-OMPA) found at high concentrations. SDS was responsible for in vitro, but not in vivo, inhibitory effects. The in vitro AChE inhibitory effects of SDS was partially prevented by the use of increasing amounts of ethanol, suggesting that the inhibition was induced by an emulsion effect, which may explain the lack of effect in vivo.     

Female and male visually based mate preferences are consistent with reproductive isolation between populations of the Lake Malawi endemic Labeotropheus fuelleborni

Sexual selection via female mate choice is thought to have played a key role in the speciation of haplochromine cichlids, but a dominant role for visual signals in such processes has lately been called into question. In addition, the possible role of male mating preferences in haplochromine speciation has been little studied. We studied patterns of both female and male mate choice, based exclusively on visual signals, in order to evaluate potential reproductive isolation between two populations of the Lake Malawi haplochromine Labeotropheus fuelleborni. In the first experiment, females were allowed to choose between two males, one from the same population and the other allopatric with respect to the female. Females in this experiment responded more frequently to males from their own population. Similarly, the males in these trials displayed more frequently when presented with females of their own population. In the second experiment, a female was allowed to choose between two males, either both from her own population or both allopatric. In these trials, both males and females from the Katale population interacted significantly more frequently in settings in which all three individuals were from the same population (same-population trios), and those from the Chipoka population showed a similar trend. Thus, patterns in both male and female courtship behavior suggest that visual signals contribute to at least incipient reproductive isolation between populations of L. fuelleborni [Current Zoology 56 (1): 65-72 2010].