Putative chemical inhibition of development by conspecifics in mosquitofish, Gambusia affinis

The objective of this study was to test for a chemical influence of adult Gambusia affinis on the development and maturation of juveniles. Sixty juveniles (<1 day old) were reared (from a total of 157 trials) individually up to 115d in 3liter containers that circulated water bathing either (1) two adult males, (2) two adult females, (3) an adult male and an adult female, or (4) conspecific-free tank water. Randomly chosen juveniles were sampled over time, sacrificed, sectioned, and measurements were taken of standard length and gonadal parameters such as gonad size, number of spermatogenic cysts, or number of oocytes. Measurements were regressed as functions of time, and analyzed for significant differences by ANCOVA. A total of 32 males and 28 females were examined. Males showed no significant differences in any of the parameters, but females showed reduced growth rate and smaller ovary size when reared in water bathing two adult females. Mortality rate in these young fish was high, but Chi-square analysis showed that mortality rate was significantly higher when they were raised in water bathing at least one adult female. Females are known to influence female development in other vertebrates; our findings suggest this may also be true for a fish, and that the interaction may be through chemical communication.     

Muscle esterases of the mosquitofish, Gambusia affinis.

1. Muscle esterases of Gambusia affinis were separable into 14 electrophoretic bands that formed five groups according to differential physical and chemical properties. 2. Nine phenotypic patterns were observed among the populations sampled, based on variation within the EST-3 group of carboxylesterases. 3. A model for the molecular and genetic bases of the EST-3 polymorphism was proposed on the basis of phenotypic patterns and physicochemical characterization. 4. The isozymes of EST-3 are inferred to be dimeric molecules on the basis of electrophoretic patterns observed following incubation of the EST-3 enzymes with neuraminidase. 5. The binding of sialic acid is believed to be an epigenetic mechanism which operates to maintain the structural integrity of these enzymes.     

Sex differentiation and pubertal development of gonads in the viviparous mosquitofish, Gambusia affinis

The ontogenetic development of gonads from embryo to adult was observed histologically in the viviparous teleost, Gambusia affinis. Primordial germ cells (PGCs) appeared in the subendodermal space of the embryo 14 days before birth, and then transferred to the dorsal mesentery to form paired genital ridges 12 days before birth. The PGCs proliferated in the genital ridge, forming gonadal primordia 10 days before birth. All gonadal primordia differentiated to the ovary containing oocytes 2 days before birth, but then redifferentiated to the ovary and testis just after birth. This indicates that the mosquitofish is a juvenile hermaphroditic species. The characteristics of gonadal sex differentiation just after birth were enlargement of the oocytes in females, and invasion of somatic cells from the hilar region to an inner portion of the gonad in males. The paired ovary fused at the basal area 5 days after birth, then on the ventral and dorsal portions, developing into a single ovary 10 days after birth. During this time a single ovarian cavity was formed on the dorsal portion of the ovary. The paired testes fused only at the basal area and became a single testis having two main lobes 10 days after birth. The oocytes gradually developed and began vitellogenesis 100 days after birth, but did not reach maturation until 110 days after birth. Spermatogenic cells formed cysts at 20 days, began meiosis at 70 days, and matured to form sperm balls 90 days after birth. The male fish sexually matured earlier than the female.     

Environmental regulation of annual reproductive cycle in the mosquitofish, Gambusia affinis

To clarify the environmental factors regulating the annual reproductive cycle of the female mosquitofish, Gambusia affinis, a viviparous teleost, histological changes of the ovary in natural population, and laboratory experiments were examined. The results, extending over two years, suggested that ovarian recrudescence is initiated by the rise in temperature during spring and that ovarian regression is caused by the shorter daylength during late summer. The first rearing experiments using four photoperiod-temperature groups to investigate the factors triggering the onset of reproduction revealed that females with regressing ovaries began reproduction with the rise of temperature regardless of the photoperiod during spring. The results of the second experiment using three different temperature groups indicated that vitellogenesis occurred at over 14 degrees C and pregnancy at over 18 degrees C. The third experiment with four photoperiod-temperature groups was arranged to investigate the factors in the cessation of reproduction. Sexually active females ceased vitellogenesis of the next clutch of oocytes due to the shorter daylength regardless of temperatures during late summer; however, temperature seemed to influence the rate of embryo development. The critical photoperiod is estimated at about 12.5 hr. In nature, it is supposed that vitellogenesis starts when the temperature rises to about 14 degrees C, and final maturation of oocytes occurs when the temperature reaches about 18 degrees C during spring. Then, vitellogenesis of the next clutch of oocytes ceases when the daylength becomes shorter than 12.5 hr during late summer; the last gestation proceeds at a rate dependent on the temperature, and finally reproduction ends by the last parturition. J. Exp. Zool. 286:204-211, 2000.     

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.     

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.     

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.     

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.     

Gonopodial morphogenesis in female mosquitofish,Gambusia affinis affinis,masculinized by exposure to degradation products from plant sterols

Synopsis Female mosquitofish,Gambusia affinis affinis, were masculinized by exposure to degradation products (presumably steroids) of the plant sterol, stigmastanol. Masculinization was indicated by the induction of a male-like gonopodium in each specimen. The morphogenetic stages of gonopodial development are discussed as are the anatomical specializations produced in the mature gonopodial tip.     

Respiratory metabolism of mosquitofish,Gambusia affinis: effects of temperature,dissolved oxygen,and sex difference

Synopsis Routine respiratory metabolic rates of mosquitofish (0.2 g live weight) were determined at 10, 15, 20, 25, 30, and 35°C at normoxic, hypoxic (= 40 torr P_O2), and extreme hypoxic (= 25 torr P_O2) conditions. Rates generally increased with increases in temperature (overall Q₁₀ = 2.11 at normoxia). Significant depressions (P<0.05) in rates were measured at 30 and 35°C at extreme hypoxic conditions. Males exhibited higher mortality at extreme hypoxia than females, especially at 35°C. Metabolic rates were significantly elevated at hyperoxic conditions (= 300 torr P_O2) at 25 and 30°C, but not at 35°C. Resting routine rates of less-confined fish were determined at 20, 25, and 30°C at normoxia and were significantly lower than the routine rates at the same temperatures. Behavioral experiments showed that aquatic surface respiration is initiated by mosquitofish at 20–65 torr P_O2 and is obligatory below 20 torr at 20° C. Overall, respiratory metabolic rates provide a quantitative metabolic basis from which predation rates of mosquitofish in various environments can be estimated.     

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.     

Tolerance of low oxygen stress in insecticide-resistant and susceptible populations of mosquitofish (Gambusia affinis)

The tolerance of low oxygen stress in insecticide-resistant and susceptible populations of mosquitofish ($\text{Gambusia}$$\text{affinis}$) was investigated. Dissolved oxygen was reduced chemically with formaldehyde and sodium sulfite treatment. Available oxygen was reduced physically through barriers, both by preventing oxygen exchange at the air-water interface and by preventing the fish from contacting the surface of the water. In all cases, the tolerance to low oxygen stress was reduced in the insecticide-resistant population compared to the susceptible population. The results indicate that although tolerance of pesticides in the resistant population has been dramatically increased by environmental selective pressures, tolerance to other stresses has not been concomitantly increased.     

Seasonal trends in body size of adult male mosquitofish,Gambusia affinis,with evidence for their social control

Synopsis Collections from a natural population of mosquitofish,Gambusia affinis, in southern Indiana showed that males born early in the breeding season mature rapidly at a small body size, while males born later in the breeding season delay maturation and achieve larger body sizes. A field experiment, involving removal of males from pond populations, was conducted to test the hypothesis that delayed maturation by late summer males is, at least in part, under social control. Mean total length (TL) of adult and maturing males in late summer in ponds from which large numbers of males had previously been removed was significantly lower than mean TL of males in control ponds. These data support the hypothesis of social control and support Sohn's competition-predation hypothesis for control of adult male body size inGambusia.     

Incorporation and utilization of multiple forms of vitellogenin and their derivative yolk proteins during vitellogenesis and embryonic development in the mosquitofish, Gambusia affinis

We previously demonstrated the presence of three forms of vitellogenin (Vg), two 600 kDa Vgs (600Vg; VgA and VgB) and a 400 kDa Vg (400Vg; phosvitinless Vg) in plasma from maturing female viviparous mosquitofish, Gambusia affinis. For further quantitative elucidation of the accumulation and utilization of the multiple Vg-derived yolk proteins, two sandwich enzyme-linked immunosorbent assays (ELISA) were developed using antisera against 600Vgs and a 400 kDa yolk protein (400Yp; derived from 400Vg), respectively. Contents of 560 kDa yolk protein (560Yp; lipovitellins derived from 600Vg) and 400Yp measured by the ELISAs increased in accordance with the growth of vitellogenic oocytes, keeping their proportional ratio (mol/mol) at about 4:1. A similar ratio obtained for plasma Vgs suggests that the proportional accumulation of the multiple Vg-derived yolk proteins is regulated by the hepatic synthesis and secretion of their precursor Vgs. When egg homogenate was analyzed by gel chromatography, three peaks, consisting of 560Yp, 400Yp and 28 kDa native beta'-component, were observed. The elution profile showed no change until embryos reached the early neurula stage, however, the relative height of the 560Yp peak as compared to the 400Yp one decreased after retinal pigmentation. Results from measurements of 560Yp and 400Yp at each embryonic stage supported the occurrence of unequal utilization of the two yolk proteins. The proportional ratios (mol/mol) of 560Yp content versus 400Yp content gradually decreased from 4.1 fold in early neurula embryo to 1.4 fold in larva just before parturition. The present study thus demonstrated unequal utilization of the multiple Vg-derived yolk proteins in developing embryos of mosquitofish.     

Effect of individual and group rearing on age and size at maturity of male mosquitofish, Gambusia affinis

Age and size at sexual maturity were significantly greater for male mosquitofish, Gambusia affinis Baird and Girard, reared in sib-groups than for their sibs reared individually. Age at sexual maturity averaged 43.3 and 62.1 days for individually- and group-reared males, respectively, a 43% difference; final length and weight at maturity were 7.7 and 34.2% greater, respectively, for group-reared males than for their individually-reared sibs. The results were consistent among 30 families that represented the progeny of 30 wild-caught females. The observed differences may be attributable to behavioural interactions affecting the neuroendocrine control of the maturation process, as suggested by previous studies of Xiphophorus .     

Deleterious effects by mosquitofish (Gambusia affinis) on the endangered fire salamander (Salamandra infraimmaculata)

Invasive fishes, via competition and predation, may be an important factor in endangering populations of amphibians and other aquatic fauna. The mosquitofish, Gambusia affinis , commonly stocked into amphibian breeding sites for mosquito control worldwide, has recently been found in some breeding sites of the endangered fire salamander Salamandra infraimmaculata in northern Israel. A comparison of Salamandra larvae in natural pools varying in Gambusia densities suggested that Gambusia negatively affects Salamandra by causing damage to its appendages, in particular, its tailfin (as manifested in a smaller tail:body ratio). Comparison of the same pool before and after mosquitofish introduction suggested strong negative effects on larval performance and number of emerging metamorphs. To explicitly test the hypothesis that Gambusia negatively affects Salamandra larval development, growth and survival, we conducted a replicated outdoor mesocosm experiment. In this experiment, we also tested how habitat structural complexity, in the form of radiating cords simulating artificial vegetation, might mediate damage caused by Gambusia . We used a two-by-two factorial design: the presence or absence of increased structural complexity crossed with the presence or absence of mosquitofish. After a short period, mosquitofish strongly reduced larval wet weight and survival, and increased body damage in the form of a reduced larval tail:body ratio and partially missing gills and limbs. No Salamandra larvae exposed to mosquitofish survived to metamorphosis in either habitat type. We conclude that Gambusia strongly and negatively affects Salamandra larvae and that the introduction of Gambusia into sites containing Salamandra is inconsistent with the goal of preserving this endangered urodele.