Evolution of the elaborate male intromittent organ of Xiphophorus fishes

Internally fertilizing animals show a remarkable diversity in male genital morphology that is associated with sexual selection, and these traits are thought to be evolving particularly rapidly. Male fish in some internally fertilizing species have “gonopodia,” highly modified anal fins that are putatively important for sexual selection. However, our understanding of the evolution of genital diversity remains incomplete. Contrary to the prediction that male genital traits evolve more rapidly than other traits, here we show that gonopodial traits and other nongonopodial traits exhibit similar evolutionary rates of trait change and also follow similar evolutionary models in an iconic genus of poeciliid fish (Xiphophorus spp.). Furthermore, we find that both mating and nonmating natural selection mechanisms are unlikely to be driving the diverse Xiphophorus gonopodial morphology. Putative holdfast features of the male genital organ do not appear to be influenced by water flow, a candidate selective force in aquatic habitats. Additionally, interspecific divergence in gonopodial morphology is not significantly higher between sympatric species, than between allopatric species, suggesting that male genitals have not undergone reproductive character displacement. Slower rates of evolution in gonopodial traits compared with a subset of putatively sexually selected nongenital traits suggest that different selection mechanisms may be acting on the different trait types. Further investigations of this elaborate trait are imperative to determine whether it is ultimately an important driver of speciation.     

Strong inbreeding depression in male mating behaviour in a poeciliid fish

The magnitude of inbreeding depression is often larger in traits closely related to fitness, such as survival and fecundity, compared to morphological traits. Reproductive behaviour is also closely associated with fitness, and therefore expected to show strong inbreeding depression. Despite this, little is known about how reproductive behaviour is affected by inbreeding. Here we show that one generation of full‐sib mating results in a decrease in male reproductive performance in the least killifish (Heterandria formosa). Inbred males performed less gonopodial thrusts and thrust attempts than outbred males (δ = 0.38). We show that this behaviour is closely linked with fitness as gonopodial performance correlates with paternity success. Other traits that show inbreeding depression are offspring viability (δ = 0.06) and maturation time of males (δ = 0.19) and females (δ = 0.14). Outbred matings produced a female biased sex ratio whereas inbred matings produced an even sex ratio.     

Within and Between Species Variation in Male Mating Behaviors in the Mexican Sailfin Mollies Poecilia velifera and P. petenensis

Polymorphism in male morphology is often correlated with the expression of alternate behavioral tactics. This relationship between behavioral and morphological polymorphisms, however, is less well understood. We characterized male mating behaviors and morphological variation within and between Poecilia velifera and P. petenensis to understand mating signal evolution in the sailfin molly lineage. In addition, we examined whether differences between these species in the size range of mature males and the strength of allometry between dorsal fin size and body length could explain the variation observed in their expression of different mating behaviors. We determined each male's mating behavior profile by observing the behavior of a single male in the presence of a receptive female. We found that P. velifera showed evidence of an alternate male mating strategy, with small males generally performing only gonopodial thrusts (forced insemination attempts) towards receptive females, while large males performed courtship displays as well as gonopodial thrusts. Males of P. petenensis performed similar rates of courtship displays and gonopodial thrusts regardless of body length. Little variation existed among populations of P. velifera in mating behaviors, while males from different populations of P. petenensis showed population‐specific average rates of each mating behavior. Population differences in P. petenensis may be driven, in part, by its occurrence in more variable habitats than those occupied by P. velifera. Variation among individuals in the mating repertoire of P. velifera, but not P. petenensis, suggests that the greater range of variation in male size at maturity, as well as considerably stronger allometry between dorsal fin size and body length, may explain why males of P. velifera show the greatest degree of expression of alternate male mating behaviors when compared to other sailfin species. These results also suggest an important role of morphological polymorphisms in predicting the expression of alternate male mating behaviors.     

Effects of 17alpha-ethynylestradiol on sexual development of male western mosquitofish (Gambusia affinis)

Juvenile male western mosquitofish (Gambusia affinis) were exposed to different concentrations of 17alpha-ethynyl estradiol (EE2) in the diet during the period of sexual maturation. A clear inhibiting influence of EE2 on sexual development was apparent. The proportion of males in each treatment group that failed to complete gonopodial development during the 150-day observation period increased significantly with EE2 concentration. There were significant nonlinear trends toward shorter gonopodia in groups exposed to higher EE2 concentrations. Vitellogenin (VTG) was detectable in the blood of all fish exposed to 1.0 or more micro/g food and the concentration increased dramatically with increasing EE2 exposure. A significant negative association was seen between EE2 concentration and spermatophore counts. This study has demonstrated deleterious effects of EE2 exposure on sexual maturation and several indirect measures of reproductive fitness. It supports the biological relevance of vitellogenin in the blood and reduced gonopodium length as biomarkers for estrogen exposure and endocrine disruption in mosquitofish.     

Individual Variation in Male Size and Behavioral Repertoire in the Sailfin Molly Poecilia latipinna

Variation among individuals in the expression of behaviors and associations of behaviors in different contexts can lead to the maintenance of behavioral polymorphisms. Individual variation in morphology is often associated with behavioral polymorphism, yet the degree to which morphology predicts behavioral phenotype is not well understood. We measured individual variation in size and behaviors in the sailfin molly, Poecilia latipinna, by comparing the behavior of individual males of different sizes across four different contexts (mating, exploratory tendency, sociability, and predator inspection). We also investigated the degree to which male size, a fixed genetic trait, influenced the expression of each behavior and associations between behaviors. We found that male mollies show strong associations between certain behaviors and only some of these are predicted by male size. For example, size has a strong influence on the courtship‐boldness axis with larger males showing higher rates of courtship displays and being bolder in predator inspection. A positive association was found between exploratory tendency, sociability, and gonopodial thrusting rates, yet the expression of these behaviors was independent of male size. Thus, sailfin mollies, like many fish species, show associations of behaviors that contribute to differences among males in personality type. The fixed genetic effect of male size at maturity influences courtship and boldness, but individual variation in exploratory tendency, sociability, and sneak copulation attempts through gonopodial thrusts is independent of male size. Such variation among males in behavioral associations within and between different contexts may slow the rate at which populations of P. latipinna can diverge in individual behaviors.     

Der Anteil des “Coolidge-Effektes” an der isolationsbedingten Zunahme sexueller Verhaltensweisen von Poecilia sphenops*)

Changing the Stimulus ♀ induced in ♂♂ of the poeciliid fish Poecilia sphenops a strong increase of copulation attempts, nipping and gonopodial swinging (“Coolidge effect”). There was a similar increase of sexual activities after social isolation of 23 hours and reintroducing the same ♀ like before. But in experiments it could be demonstrated that the effect of social deprivation can only partly be deduced from the Coolidge effect and seems to be also the result of sexual inactivity during the isolation period.     

Gonopodial system review and a new fish record of Poeciliopsis infans (Cyprinodontiformes: Poeciliidae) for Lake Patzcuaro,Michoacan, central Mexico

Since 1997, Poeciliopsis infans Woolman 1894 has been recognized as a new inhabitant of Lake Patzcuaro, Michoacan in Central Mexico. Between February 1997 and October 1998, nine fish samples were collected at Lake Patzcuaro. Morphometric and meristic counts were conducted on a random selection of 40 organisms of both sexes of Poeciliopsis infans. Males of these viviparous fish posses a modified anal fin called gonopodium. The characteristic hemal spine on the 18th caudal vertebra for this species is described herein and the bony components of the gonopodial structure and suspensoria that together comprise the gonopodial system, which is important for taxonomic studies at various levels of classification were reviewed. Poeciliopsis infans displays a high degree of sexual dimorphism in body shape and anal fin anatomy with the most conspicuous difference observed in anal fin height, which averages 40% of SI in males and 17% in females. Comparisons between male and female anal fins are described herein as well as the possible impacts of this species on Lake Patzcuaro fish fauna.     

Polymorphism and pair formation in the mosquitofish Gambusia holbrooki (Pisces: Poeciliidae)

Synopsis Pair formation was studied in a polymorphic population of Gambusia holbrooki in a brackish drainage canal in Southern Florida. All females and the majority of males were gray colored and only 5% of the males were black spotted. A small fraction of the population formed pairs, composed of a female and a male that swam synchronously and close together for several minutes. During pairing the male often achieved gonopodial contact with the female. Black spotted males occurred in pairs ten fold more frequently than expected under random pairing. The major behavioral mechanism accounting for the disproportionate occurrence of spotted males in pairs is, probably, the preference of females for these males.     

Seasonal Abundance and Reproduction of the Fourwing Flyingfish, Hirundichthys affinis, off Barbados

The seasonal abundance and reproductive cycle of the flyingfish Hirundichthys affinis was studied near Barbados. H. affinis shows large seasonal variation in abundance. They are common between December and June and scarce from July to November. The population size structure of H. affinis was determined by fishing with four different mesh sizes throughout one year. There appear to be two cohorts of H. affinis present in May and June, one of immature fish with a mean size of 19.8cm FL, and one of mature fish with a mean size of 21.7cm FL. From September to April there is only a single cohort of maturing fish with a mean size that increases from 20.4cm FL in September to 22.0cm FL in April. H. affinis spawns throughout the fishing-season (December–June) and shows a peak in spawning activity from March to June. Immature fish are present in Barbados waters from July to November but in very low abundance. Surface water temperature, wind speed and swell height are the only environmental factors with which catch rate of H. affinis showed significant correlation. Catch rate appears to be negatively correlated with temperature, whereas for wind speed and swell height there are optima. Three hypotheses for the seasonal variation in abundance of H. affinis were examined. The hypothesis that this species remains dispersed near Barbados during the off-season was rejected. The other hypotheses were refined. Both post-spawning mortality, resulting in an interval of low abundance between cohorts (non-overlapping generations), and migration of fish away from Barbados at the end of the fishing season, remain possible explanations for the observed seasonal variation in abundance of this species.     

Retinoic Acid Is Involved in the Metamorphosis of the Anal Fin into an Intromittent Organ,the Gonopodium,in the Green Swordtail (Xiphophorus hellerii)

In poeciliid fish the male anal fin has been transformed into a gonopodium, an intromittent organ required for internal fertilization. Elevated testosterone levels induce metamorphosis of a subset of anal fin rays to grow and form the specialized terminal structures of the gonopodium. The molecular mechanisms underlying these processes are largely unknown. Here, we investigated whether retinoic acid (RA) signaling is involved in gonopodium development in the swordtail Xiphophorus hellerii. We showed that aldh1a2, a RA synthesizing enzyme, and the RA receptors, rar-ga and rar-gb, are expressed in anal fins during metamorphosis. aldh1a2 expression is regulated by testosterone in a concentration-dependent manner and is up-regulated in both hormone-induced and naturally developing gonopodia. Androgen receptor (ar), a putative regulator of gonopodial development, is co-expressed with aldh1a2 and the RA receptors in gonopodial rays. Importantly, experimental increase of RA signaling promoted growth of the gonopodium and increased the number of new segments. Based on gene expression analyses and pharmacological manipulation of gonopodium development, we show that the RA signaling pathway is activated in response to androgen signaling and promotes fin ray growth and development during the metamorphosis of the anal fin into the gonopodium.     

Males,but not females,contribute to sexual isolation between two sympatric species of <Emphasis Type="Italic">Gambusia</Emphasis>

Reproductive isolation restricts genetic exchange between species. Various pre- and post-mating barriers, such as behavior, physiology and gametic incompatibility, have been shown to evolve in sympatry. In certain scenarios, isolation can be asymmetrical, where species differentially prefer conspecifics. We examined sexual isolation via conspecific mate preference between Gambusia affinis and G. geiseri in both sexes. To investigate male contribution to sexual isolation, we compared the number of mating attempts (gonopodial thrusts) directed at either a conspecific or a heterospecific female, in both species. We also examined sperm priming and expenditure in males in the presence of conspecific or heterospecific females. We then measured female preference for either a conspecific or heterospecific male, in both species. We found that males of both species preferred to mate with conspecific females, but showed no difference in sperm production or expenditure between conspecific and heterospecific females. Females of both species did not prefer conspecific over heterospecific males. Our results suggest that sexual isolation might be mediated by male mate choice in this system and not female choice, suggesting that there is asymmetrical reproductive isolation between the sexes in G. affinis and G. geiseri, but symmetrical species isolation.     

Population development of the mosquitofish,Gambusia affinis,in rice fields

Synopsis Development of mosquitofish,Gambusia affinis, stocks in rice fields following stocking for mosquito control is poorly understood and highly variable. To characterize population development and explain observed variability, size distributions and total numbers of mosquitofish stocks were followed from stocking through the end of the summer rice season in several experimental rice paddies. Instead of the highly variable, logistic growth implied by previous studies in which only adult fish were sampled, we observed a consistent development of size structure between years, with some variability in specific demographic processes. Population development consists of: (1) an initial peak in numbers due to a pulse of recruitment, (2) a period of low recruitment and constant or declining numbers, and (3) a second peak caused by a second pulse of recruitment. Timing of the initial peak appears to depend on rice height. Differences in the rate of decline following the peak are apparently due to different mortality rates. The second peak in reproduction is probably due to a second parturition by stocked females, possibly followed by first reproduction by young born earlier. Since population development in rice fields following stocking in the spring is similar to development of natural populations in temperate regions in which there is no reproduction during the winter, results obtained here are relevant to natural as well as artificially stocked systems.     

Phenology of Dryopteris affinis ssp. affinis and Polystichum aculeatum: modeling relationships to the climatic variables in a Mediterranean area

The phenology of leaves and stages from immature sori to spore release were studied in natural populations of two perennial herbaceous ferns, Dryopteris affinis ssp. affinis and Polystichum aculeatum, for 2 years in Italy. The Generalized Additive Model for Location, Scale and Shape (GAMLSS) was used to predict a particular phenological event from the climatic variables. Both fern species are evergreen with leaf lifespan of 13–14 months in D. affinis ssp. affinis and 15–24 months in P. aculeatum. Their leaf production is seasonal with most flushed in spring. In D. affinis ssp. affinis the decaying of old leaves is sudden whereas in P. aculeatum it is gradual, and in both types the decaying is accelerated just as the new leaves emerge. Temperature is the best predictor for the development of sori and spore release. The end of spore release and leaf emergence are positively affected by temperature and negatively affected by rainfall and snow cover for P. aculeatum. An almost similar response to climatic factors of emergent and senescent leaves and their phenology supportsthe hypothesis that old leaves serve as nutrient storage organs for new leaf growth. Comparison the phenological patterns between the 2 years indicated that the time lapses between each phenological event were the same within seasons for D. affinis ssp. affinis but show some differences for P. aculeatum. We also hypothesize that endogenous factors may play an important role in the phenology of P. aculeatum.     

The effects of induced masculinization on reproductive and aggressive behaviors of the female mosquitofish,Gambusia affinis affinis

Synopsis FemaleGambusia affinis affinis were masculinized with the degraded products of 65% stigmastanol-30% B-sitosterol, a phytosterol. The masculinized females were paired with non-treated males, non-treated females, and other masculinized females. The pairs were analyzed for reproductive and aggressive behaviors exhibited. The behavioral patterns of these pairs were compared statistically to the behavior patterns of the following non-treated pairs: male with male, female with female, and male with female. The masculinized females behaved like males in that they followed, swung, and thrust their gonopodia at non-treated females and larger masculinized females. However, the male-like behavior of the masculinized females was not as intense as that of normal males. Also, the effect of masculinization on the behavior of the treated female was context-dependent as seen when placed with a male or a smaller masculinized female. Under these conditions the masculinized females behaved like typical females and exhibited no masculinized behaviors. The effects of anal spots and size differences are discussed as possible explanations for the variability in behaviors exhibited. The masculinized females displayed no change in aggressive behaviors.     

Influence of tissue culture conditions on apogamy in Dryopteris affinis sp. affinis

The naturally-occurring apogamy of some ferns can be modified by culture conditions and growth regulators. Gametophytes of the apogamic fern Dryopteris affinis sp. affinis L., were cultured on Murashige and Skoog (MS) basal medium. Changes in concentration of MS medium components, sucrose, agar and different pH values were tested. The addition of benzyladenine (4.43 M) and naphthalene acetic acid (0.53 M) enhanced sporophyte proliferation on the gametophytes. After one month in culture, the gametophytes formed callus with a high morphogenic capacity. Culture of calli on medium without growth regulators yielded about 10,000 sporophytes per 1 g fresh weight of callus. This pattern of differentiation slowed with time to a point where only gametophyte regeneration was observed.Abbreviations BA benzyladenine - 2,4-d 2,4-dichlorophenoxyacetic acid - F.W. fresh weight - MS Murashige & Skoog medium - NAA 1-naphthalene acetic acid - SE standard error     

Rapid responses to stress in Eurytemora affinis

E. affinis can adjust to temperature stress in a matter of hours. Adaptation is greater in a varying temperature than in a constant temperature, consistent with the estuarine habitat of this calanoid. The species has the capacity to adjust both in the short-term as individuals and also genetically over a number of generations. The adjustments have been examined at several levels of organization. In whole copepods the time an individual becomes comatose when exposed to a 32 °C temperature and increasing by 1/2 °C at 5 min. intervals, has been used as a repeatable assay and gives a good prediction of survival at 30 °C, the ecological limit of the species in Chesapeake bay, USA. At the molecular and cellular levels, two adaptive mechanisms which have been observed in temperature stressed copepods are the synthesis of novel proteins and phase changes in plasma membrane lipids. Both of these mechanisms have potential for further understanding the adaptation of Eurytemora to variable temperatures. They may also have application as indicators of sublethal stress.     

Allometric relationship between oxygen consumption and body weight of mosquitofish,Gambusia affinis

Synopsis The allometric relationship between body size and oxygen consumption of Gambusia affinis at 28° C was determined under controlled experimental conditions, using a manometric respirometer. The allometric exponent (b-value) was 0.64 ± 0.02 S.E. (n = 51). Oxygen consumption was not influenced by any time-related factors during the 17 days of measurements. Variance between replicated oxygen consumption trials on individual fish was negligible. Specific oxygen consumption rates of several G. affinis at 28° C that were determined by using a sealed vessel and O₂ electrode respirometry method were similar to the rates measured by the manometric respirometry method in similar-sized G. affinis.     

Impacts of mosquitofish (Gambusia affinis) predation on plankton communities

An investigation of the effects of mosquitofish (Gambusia affinis) predation was conducted in 12 experimental ponds in southern California over a period of 10 months.Gambusia essentially eliminatedDaphnia pulex andCeriodaphnia sp. populations, reducedDiaptomus pallidus andKeratella quadrata populations, had little impact onCyclops vernalis, and caused large increases inK. cochlearis, Polyarthra sp.,Synchaeta sp., andTrichocerca spp. populations and in total phytoplankton.Gambusia caused a decrease in the PIE (probability of interspecific encounter) of the planktonic crustaceans and an increase in the PIE of the planktonic rotifers. Hemiptera, such as neustonicMicrovelia sp. and nektonicBuenoa sp. andNotonecta sp., andHyla regilla tadpoles were absent from fish ponds but sometimes abundant in control ponds.Gambusia caused higher pH and oxygen levels, presumably via its effect on the phytoplankton. The impact ofGambusia on the pond ecosystems was less in winter, when fish numbers and feeding rates were low, than in summer. Results of other fish-plankton studies are summarized in tabular form. A model is proposed to account for variation in the calanoid/cyclopoid ratio; evidence is summarized suggesting that in general calanoids are more susceptible to predation by predaceous zooplankters while cyclopoids are more susceptible to fish predation. Some parallels are drawn between the effects ofGambusia predation and those of insecticide treatments.