Domperidone treatment attenuates stress-induced suppression of reproduction in viviparous mosquitofish Gambusia affinis

The aim of this study was to determine the effect of stress on reproduction and the possible involvement of dopaminergic systems in the reproductive stress response in the mosquitofish Gambusia affinis. Exposure of fish to aquaculture stressors (four 10 min episodes of stress, each corresponding to a different stressor such as handling, chasing, frequent netting and low water levels), for a period of 30 days caused reduction in the mean numbers of stage I–IV follicles associated with lower number of pregnant females and embryos in most of the developmental stages compared with experimental controls. Besides, increase in the intensity of labelling and the per cent area of tyrosine hydroxylase (TH; a rate-limiting enzyme in the biosynthetic pathway of catecholamines)- immunoreactive (ir) neurons was observed in the preoptic area (POA) and the nucleus preopticus (NPO) regions of the brain concomitant with reduction in the labelling of gonadotropin releasing hormone–immunoreactive (GnRH-ir) fibres in the proximal pars distalis (PPD) of the pituitary gland in stressed fish compared with experimental controls. Treatment of domperidone (DOM) caused an increase in the number of stage II and V follicles and promoted pregnancy rate concomitant with an increase in the number of embryos at various developmental stages compared with those of experimental controls. Similar treatment to stressed fish caused an increase in the number of stages I–V follicles compared with those in stress alone group. The GnRH fibres showed increased immunolabelling in stress + DOM treated fish compared with stress alone fish. On the other hand, TH-immunoreactivity in the POA and the NPO regions was reduced in stress + DOM treated fish compared with stress-alone group. These results suggest that stress inhibits follicular development and subsequent hatching success through the suppression of GnRH and that the inhibition appears to be mediated through dopamine, for the first time in a viviparous fish.     

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.     

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.     

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.     

Geographical and temporal variation of multiple paternity in invasive mosquitofish (Gambusia holbrooki,Gambusia affinis)

Multiple paternity (MP) increases offspring's genetic variability, which could be linked to invasive species' evolvability in novel distribution ranges. Shifts in MP can be adaptive, with greater MP in harsher/colder environments or towards the end of the reproductive season, but climate could also affect MP indirectly via its effect on reproductive life histories. We tested these hypotheses by genotyping N = 2,903 offspring from N = 306 broods of two closely related livebearing fishes, Gambusia holbrooki and Gambusia affinis. We sampled pregnant females across latitudinal gradients in their invasive ranges in Europe and China, and found more sires per brood and a greater reproductive skew towards northern sampling sites. Moreover, examining monthly sampling from two G. affinis populations, we found MP rates to vary across the reproductive season in a northern Chinese, but not in a southern Chinese population. While our results confirm an increase of MP in harsher/more unpredictable environments, path analysis indicated that, in both cases, the effects of climate are likely to be indirect, mediated by altered life histories. In both species, which rank amongst the 100 most invasive species worldwide, higher MP at the northern edge of their distribution probably increases their invasive potential and favours range expansions, especially in light of the predicted temperature increases due to global climate changes.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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 .