A Review of the Biology of <Emphasis Type="Italic">Gambusia affinis</Emphasis> and <Emphasis Type="Italic">G. holbrooki</Emphasis>

Eastern and Western Gambusia (i.e., Gambusia holbrooki and G. affinis, respectively) are considered together here because these two fish species are very closely related, similar in appearance, similar in biology and often confused. Widely divergent attitudes have developed with regard to these fish with some viewing them as being highly beneficial to humans through controlling mosquitoes and the diseases they harbor, and others expressing concern about the negative impacts that these fish may have on other species with which they interact. Because of the widespread distribution, high levels of abundance, ease of capture and captive maintenance, and divergent attitudes, a very large and diffuse literature has developed with regard to these species. In fact, few fish species have been studied as much as or more than these two species combined. There has, however, been no comprehensive review of their biology published to date. As it is not possible to provide a comprehensive review of Gambusia biology in one reasonably sized document, I provide here a review of aspects of their biology at the level of species and individual. In another review I focused instead on the levels of population and species communities and consider the impacts that these fish have on mosquitoes and other organisms (Pyke, unpublished). As would be expected of such widespread and abundant species, Gambusia affinis and G. holbrooki are clearly very tolerant, adaptable and variable in their biology, at both an individual and population level. Both individuals and populations can tolerate, and often thrive within, a wide range of conditions and the abilities of individuals to do this are enhanced if they have time to acclimate to any changes. Populations can adapt through genetic or evolutionary changes in response to conditions that vary in space or time, and there is significant genetic variation within and between populations.     

Sex Differences in Mate Preference Between Two Hybridizing Species of Poeciliid Fish

When hybridization between two species in secondary contact is costly, natural selection should favor pre‐mating isolation barriers. The invasive Gambusia affinis has been introduced to habitats of a closely related species, the endangered Gambusia nobilis. Although other Gambusia species readily hybridize in secondary contact, previous studies in this system found low abundance of hybrids in sympatry. To examine whether hybridization is limited by behavioral pre‐mating isolation that may have evolved in allopatry, I examined each species’ mating preferences using individuals from allopatric populations in male and female visual/olfactory association preference tests as well as open mating tests with and without male–male competition. Gambusia affinis and G. nobilis males had significant association preference for conspecific females in visual/olfactory tests. Only G. nobilis females had statistically significant preference for conspecific males. In open mating tests, males of both species had lower chase times overall when in competition, but there was no difference in number of copulation attempts. Males of both species had higher copulatory success rates with conspecific females when in competition, suggesting females may exert some control over copulation success of males. These results suggest that there are differences in mating preferences between these species. This mate choice may act as a pre‐mating isolating barrier to reduce hybridization in sympatry, a proposed threat to the endangered G. nobilis.     

HYBRID ZONE DYNAMICS ARE INFLUENCED BY GENOTYPE-SPECIFIC VARIATION IN LIFE-HISTORY TRAITS: EXPERIMENTAL EVIDENCE FROM HYBRIDIZING GAMBUSIA SPECIES

Results from two experiments are presented that contrast differences in life-history traits and population dynamics between two species of live bearing fishes (Gambusia affinis and G. holbrooki) that hybridize across portions of the southeastern United States. Progeny from parental holbrooki and holbrooki-affinis F₁ crosses exhibited larger lengths at birth, at 15 days, and matured earlier, and at larger size than did progeny from parental affinis and affinis-holbrooki F₁ crosses. Comparisons of experimental populations of affinis, holbrooki, and mixed (affinis + holbrooki) species composition followed over two years revealed that affinis populations consistently exhibited smaller population size, lower carrying capacity, lower recruitment, and larger over-winter mortality than did holbrooki or mixed populations. Evidence for density-dependent reductions in fecundity and concomitant increases in juvenile mortality rates were observed in all populations, but were most pronounced for affinis populations. Genotype-specific differences in life-history traits appear to confer differential advantage to offspring of parental holbrooki origin and F, progeny of holbrooki maternal parentage given the resource availability and the age structure and densities experienced during these experiments. Results have direct implications regarding the rate and direction of evolution within hybrid zones formed by these two species.     

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.     

Empirical evaluation of cytonuclear models incorporating genetic drift and tests for neutrality of mtDNA variants: data from experimental Gambusia hybrid zones

Statistical tests of genetic drift and of the neutrality of mtDNA are presented using empirical time‐series data on multi‐generational changes in cytonuclear disequilibria within replicated experimental hybrid populations of two species of live‐bearing Poeciliid fishes (Gambusia holbrooki and G.affinis) which were monitored over a period of two years (three generations). Cytonuclear disequilibria D and D (which measure departures from random associations of cytoplasmic and nuclear genotypes) over the three generations of the experiment were non‐zero for all replicate populations. For each of five nuclear loci, the observed measures of D and D were highly concordant between replicates during each generation. Significant departures from expectations were observed after one and two generations. A statistical measure of goodness of fit of observed changes in cytonuclear disequilibria (and implicitly of the neutrality of the mtDNA markers) was calculated for each nuclear locus. When the results for the replicates were combined into an overall test of neutrality, the fit to the random union of zygotes (RUZ) model was rejected for four of the five nuclear loci (P < 0.05). A simple genetic drift model does not explain the temporal changes in composite cytonuclear genotypic frequencies. Frequencies of parental G. holbrooki mitochondrial alleles and nuclear genotypes exceeded expected values during most time periods, implying some selective advantage of offspring produced by G. holbrooki females. Expansion of cytonuclear models to explicitly address questions of genetic drift and neutrality have general relevance to studies of natural populations. This revised version was published online in July 2006 with corrections to the Cover Date.     

Gambusia quadruncus (Cyprinodontiformes: Poeciliidae): a new species of mosquitofish from east‐central México

Gambusia quadruncus n. sp., the llanos mosquitofish, is described from east‐central México. The region inhabited by the species represents a hotspot of diversity of Gambusia, and G. quadruncus sometimes coexists with at least three congeners. The species differs from its closest relative, Gambusia affinis, in several characteristics with plausible effects on reproductive isolation, e.g. body size, body and fin morphology, male genital morphology (distal tip of gonopodium) and female anal spot morphology (colouration near the urogenital sinus). Moreover, combined analysis of mitochondrial and nuclear gene sequence data (c. 2158 total base pairs) indicates reciprocal monophyly of G. quadruncus and its sister species G. affinis, with levels of genetic divergence suggesting the two species diverged from one another over a million years ago. The origin of G. quadruncus may reflect a vicariant event associated with Pliocene orogenesis in the Tamaulipas Arch and a frontal section of the Sierra Madre Oriental (Lleran Mesas). Gambusia quadruncus inhabits a variety of freshwater habitats across several river drainages, with its range spanning at least 350 km from north to south, covering over 25 000 km². A key to aid identification of the species is provided.     

Diversity and fine-scale spatial genetic structure of Cyperus papyrus populations in Lake Naivasha (Kenya) using microsatellite markers

Species introduced outside their native range to serve anthropogenic purposes may have unintended consequences on native organisms. Mosquitofish (Gambusia affinis and Gambusia holbrooki) have been introduced throughout the world to control larval mosquito populations in aquatic environments, but they have also been implicated in the decline of native fish, amphibian, and aquatic invertebrate populations. We investigated the roles of introduced western mosquitofish (G. affinis), and two naturally colonizing predators (crayfish and dragonfly naiads) in inflicting tail injuries observed on ranid frog tadpoles in experimental constructed wetlands. We also examined the influence of vegetation in reducing tail injury severity. We found that mosquitofish significantly increased the prevalence and severity of tail injuries, but crayfish and dragonfly naiads contributed much less to increased injury levels. Furthermore, increased vegetation cover did not significantly attenuate tail injuries. However, after chemical removal of Gambusia, injury prevalence and severity was significantly reduced. Although previous investigations have concluded that mosquitofish prey upon the eggs and larvae of some amphibians, our results illustrate that these fish can cause substantial but apparently sub-lethal injuries to large numbers of larval amphibians in a wetland. Further investigations are needed to determine if these injuries impede the fitness of victims and lead to population reductions.     

Cytonuclear genetic architecture in mosquitofish populations and the possible roles of introgressive hybridization

Spatial genetic structure in populations of mosquitofish ( Gambusia ) sampled throughout the south-eastern United States was characterized using mitochondrial (mt) DNA and allozyme markers. Both sets of data revealed a pronounced genetic discontinuity (along a broad path extending from south-eastern Mississippi to north-eastern Georgia) that corresponds to a recently recognized distinction between the nominal forms G. affinis to the west and G. holbrooki to the east. However, several populations from the general contact region exhibited unusual allelic associations in high frequency, suggestive of evolutionary processes within a zone of introgressive hybridization. These involve: (i) cytonuclear profiles representing combinations of nuclear and mitochondrial genotypes that tended to be more nearly species-specific and concordant elsewhere; and (ii) significant nuclear gametic disequilibria, perhaps attributable to positive assortative mating and/or differential fitnesses of homospecific vs. recombinant genotypes. However, outside this suspected hybrid region, 'heterospecific' genetic markers also appeared in low frequency, thus complicating interpretations. These discordant alleles on a broader geographic scale may reflect: (a) the retention of polymorphisms from an ancestral gene pool; (b) occasional evolutionary convergence (especially with respect to electrophoretic mobility of allozyme alleles); (c) the 'footprints' of a moving hybrid zone; or (d) differential introgressive penetrance across the current hybrid region.     

Dispersal Behavior, Boldness, and the Link to Invasiveness: A Comparison of Four Gambusia Species

Dispersal is a key element of a species' invasiveness. Although considerable work has addressed how dispersal influences the pattern of spatial spread of invading organisms, few studies investigate whether invasive species are in fact better dispersers than either the species they displace or less successful invaders. Recent work suggests that variation in dispersal may be due to variation in an underlying behavioral trait, boldness. Our study examined the link between dispersal, boldness, and invasiveness by comparing the dispersal characteristics and refuge use of two invasive Gambusia species to two congeners in experimental streams. The streams consisted of a series of pools (no flow) connected to a flowing channel. For each species, small groups of females were released at the middle pool, and their movement and activity were recorded over a 1-h period. We found invasive Gambusia to be more likely to disperse out of the introductory pool, to disperse sooner, to travel a greater distance in the artificial streams, and thus to exhibit greater dispersal tendencies than their close relatives. Among the invasives, Gambusia affinis had a greater dispersal tendency than G. holbrooki. We suspect this result indicates variation in the contribution of dispersal to the relative invasiveness of these species. Certain dispersal measures were correlated to time spent out of refuge, although invasive Gambusia and their relatives did not differ in the predicted manner. These results argue for the greater incorporation of experimental approaches and analyses of behavioral mechanisms in the study of invasive species.     

Microsatellite assessment of multiple paternity in natural populations of a live-bearing fish,Gambusia holbrooki

Three polymorphic microsatellite loci were isolated and employed to examine paternity patterns in two natural populations of live-bearing mosquitofish, Gambusia holbrooki. Each locus displayed four to five alleles per population in samples of presumably unrelated adults. Nearly 900 embryos from a total of 50 pregnant females were assayed individually, and paternal alleles in each embryo were identified. Counts of paternal alleles, Mendelian segregation patterns, multilocus allelic associations and genetic relatedness coefficients were employed to estimate the minimum and effective numbers of fathers per brood. At least 90% of the assayed broods were shown to have been fathered by multiple males, a figure substantially higher than previous estimates based on less polymorphic genetic loci. However, the genetic data yield a face-value estimate of only about 2.2 fathers per brood, a number that seems perhaps surprisingly low based on frequencies of attempted copulations by males. Both biological and sampling factors that might bias mean sire counts downward are considered. Although higher sire counts per brood might be obtained from loci with even greater numbers of alleles, little statistical room remains for higher frequency estimates of multiple paternity in Gambusia.     

Response of endangered Poeciliopsis occidentalis sonoriensis in the Río Yaqui drainage,Arizona, to introduced Gambusia affinis

Synopsis Potential coexistence of the native Yaqui topminnow, Poeciliopsis occidentalis sonoriensis, with introduced mosquitofish, Gambusia affinis, was examined in spring pools and streams in San Bernardino National Wildlife Refuge, southeastern Arizona. Poeciliopsis never exceeded 12% of total poeciliid numbers in a spring pool where Gambusia was present. Body size, fecundity and reproductive effort of Poeciliopsis were significantly higher in this pool than in similar spring pools where Gambusia was absent. Where Poeciliopsis and Gambusia were syntopic in a stream, numbers of Poeciliopsis declined over 1.7 years, until none were collected. Poeciliopsis then increased to > 60% of total poeciliid numbers following flash flooding. Persistence of Poeciliopsis with Gambusia in the spring pool appeared to be a result of compensatory increase in reproductive output, while in the stream it was associated with recurrent flash flooding and a uniform temperature springhead which provided refuge.     

The establishment of the North-American mosquito fish,Gambasia holbrooki,in Syrian inland waters)

Abstract

The North-American mosquito fish, Gambusia holbrooki (Girard, 1859), came to Syria in 1929 but only recently spread over large parts of the country. Between 1977 and 1979 Gambusia was studied in various water bodies throughout Syria. The conditions For its successful establishment and its effect on populations of the indigenous cyprinodont fish Aphanius mento (Heckel, 1843) were surveyed. Out of 31 localities where Gambusia and/or Aphanius occurred 17 were inhabited by both species, 11 only by Aphanius and three only by Gambusia. A comparison of these localities indicated that Gambusia is better adapted to high salinity and polluted water whereas Aphanius tolerates stronger currents. Possible negative effects of Gambusia on the indigenous aquatic fauna of Syria are briefly discussed.     

Effects of maternal and embryo characteristics on post-fertilization provisioning in fishes of the genus <Emphasis Type="Italic">Gambusia</Emphasis>

Maternal provisioning of embryos in Gambusia (Poeciliidae) entails both production of large, yolky eggs and mother-to-embryo transfer of nutrients, the latter of which is readily quantified using injection of radiolabeled nutrients. We assayed patterns of nutrient transfer in broods of 26 Gambusia geiseri and 23 Gambusia affinis females, using injection of tritiated leucine. We examined maternal and embryo characteristics affecting the instantaneous rate of transfer and characterized the pattern of transfer to individual embryos within broods. Maternal (female size and condition) and brood characteristics (mean embryo size, developmental stage, brood size) did not predict the mean level of nutrient transfer to embryos in a brood for either species. Within broods, individual provisioning of embryos was not related to developmental stage, but was related to embryo mass in G. affinis with nutrient transfer higher to larger embryos. In addition, overall within-brood variation in nutrient transfer, measured as coefficient of variation in embryo radioactivity, was higher in G. affinis than in G. geiseri.     

An invasive fish species within its native range: community effects and population dynamics of Gambusia affinis in the central United States

1. Management of invasive species benefits from detailed information on the biology of the invaders, both from where they have already invaded, and from within their areas of origin. Western mosquitofish, Gambusia affinis, is a widely invasive and destructive freshwater fish. However, within its native range, G. affinis co‐exists with many other fish species in a wide variety of habitats without obvious harm. 2. In this study, we used data on fish communities within the native range of G. affinis at 154 sites across a broad spatial scale to examine the effects of G. affinis on species richness and diversity of residual (species other than G. affinis) fish assemblages. We further used data based on annual samples at eight fixed river sites over 18 summers to examine temporal population dynamics of G. affinis and to test factors associated with population fluctuations. 3. Higher residual species richness occurred in the presence of G. affinis, but residual diversity did not differ. We found an inverse relationship between relative abundance of G. affinis and residual species richness (although effect size was extremely small), but no effect on residual diversity. 4. Gambusia affinis populations fluctuated markedly across summers at all eight fixed sites, but population sizes at a site over time were not autocorrelated. However, population fluctuations were highly correlated among sites across all years, suggesting that regional factors influenced population size. Regional abundance of G. affinis did not correlate with drought, rainfall or winter temperature, but varied with spring temperature. We suggest earlier onset of reproduction in warmer springs resulted in larger summer populations. 5. Overall, within its native range, G. affinis does not appear to impact negatively on the assemblages in which it occurs, possibly due to fluctuations in its density. These findings suggest that introduced Gambusia populations, and those of other invasive species, warrant careful monitoring over long periods of time where they have invaded. Long‐term monitoring of new populations can establish if they are prone to ‘boom and bust’ dynamics, in which case the invader may be less a threat than sometimes assumed. Population information from long‐term studies, either in their native ranges or at invaded sites, can thus help to form the basis of prudent, cost‐effective management strategies for invasive organisms.     

Community level impacts of invasive mosquitofish may exacerbate the impact to a threatened amphibian

Invasive fish threaten many native freshwater fauna. However, it can be difficult to determine how invasive fish impact animals with complex life cycles as interaction may be driven by either predation of aquatic larvae or avoidance of fish‐occupied waterbodies by the terrestrial adult stage. Mosquitofish (Gambusia spp.) are highly successful and aggressive invaders that negatively impact numerous aquatic fauna. One species potentially threatened by Gambusia holbrooki is the green and golden bell frog (Litoria aurea). However, G. holbrooki's role in this frog's decline was unclear due to declines driven by the chytrid fungal disease and the continued co‐existence of these fish and frogs in multiple locations. To clarify the extent to which Gambusia is impacting L. aurea, we conducted 3 years of field surveys across a deltaic wetland system in south‐east Australia. We measured the presence and abundance of aquatic taxa including G. holbrooki, and L. aurea frogs and tadpoles, along with habitat parameters at the landscape and microhabitat scale. Generalized linear models were used to explore patterns in the abundance and distributions of L. aurea and G. holbrooki. We found strong negative associations between G. holbrooki and tadpoles of most species, including L. aurea, but no apparent avoidance of G. holbrooki by adult frogs. Native invertebrate predators (Odonata and Coleoptera) were also absent from G. holbrooki‐occupied ponds. Due to the apparent naivety of adult frogs toward G. holbrooki, the separation of G. holbrooki and tadpoles, plus the abundance of alternative predators in G. holbrooki‐free ponds, we conclude that the impact of G. holbrooki on L. aurea recruitment is likely substantial and warrants management action.     

Genetic characterization of the invasive mosquitofish (Gambusia spp.) introduced to Europe: population structure and colonization routes

Biological invasions are considered one of the main anthropogenic factors that reduce the abundance of native species. Understanding the patterns of population structure and behavior of introduced species is important to determine invasion sources and pathways, in addition to improving the protective management of native species. Thus, we set out to advance our knowledge about the mosquitofish Gambusia spp., which is an invasive species that was introduced to southern Europe in 1921 to control mosquito populations. We assessed the genetic diversity and population structure of this species at 13 European locations, by screening variation at six microsatellite loci. We also evaluated six American samples (four of G. holbrooki and two of G. affinis) to identify the most likely source of the populations that established in Europe, and to determine whether G. affinis is also present. The results showed that, while there was evidence of recent bottleneck events in a few isolated locations, most introduced populations harbored a considerable amount of gene diversity, probably because of multiple introductions and secondary contacts. Populations displayed strong genetic differentiation that was mainly associated with geographical distance. At least two main routes of colonization of G. holbrooki seem to have occurred in Europe. The first, and more ancient colonization, was consistent with historical records, with the species invading the Iberian Peninsula. A second and more recent colonization probably occurred in Greece and, from there, France. The presence of G. affinis was not detected in any of the European samples.     

Does seasonal flooding give a native species an edge over a global invader?

1. The factors that promote coexistence of global invaders and native species are poorly understood. The role of abiotic factors in determining the dominance of invasive species is also an area of increasing interest. Gambusia holbrooki (Eastern Mosquitofish) is an important global invader, displacing endemic fish and frogs on four continents. However, Gambusia co‐occurs with a similar‐sized fish (Hypseleotris spp.) in dynamic floodplain wetland pools of south‐eastern Australia. 2. We examined the relative abundance of Gambusia and Hypseleotris in regularly flooded and irregularly flooded pools to determine whether seasonal hydrological disturbance may be advantageous to the native species. We aimed to determine whether Gambusia and Hypseleotris populations respond differently to regular seasonal flooding and whether this could reduce the ability of Gambusia to dominate numerically the native species. We tested this by sampling fish bi‐monthly in 15 floodplain pools over 2.5 years. 3. We found that the relative abundance of the two species differed between regularly and irregularly flooded pools, while both pool types persisted (over a period of a year). Hypseleotris were numerically dominant in regularly flooded pools from spring through to autumn, following a major spring flood. In contrast, Gambusia were more abundant than Hypseleotris in irregularly flooded pools from summer until pools dried completely. 4. Due to the higher reproductive response of Hypseleotris to a spring flood, and the timing of its breeding season, we suggest that Hypseleotris may have a reproductive advantage over Gambusia during productive post‐flood spring/summer conditions. While the abundances of both species were usually within a similar range, variation in hydrologic habitats and the inter‐play between life‐history traits and hydrologic disturbance may nonetheless give the native fish an edge.     

Biochemical genetics of mosquitofish III. Incidence and significance of multiple insemination

Evidence for multiple insemination in the mosquitofish, Gambusia affinis, was detected by statistical analysis of the enzyme phenotypes in mother-offspring combinations for three loci. The probability of nondetection of multiple insemination events for diallelic loci with two or more males mating with a female was presented. The frequency of multiple insemination for G. affinis is at least 56% but is probably nearer to 100%. The high incidence of multiple insemination together with sperm storage and delayed fertilization could have important effects on allele frequencies through time and space. These effects are due to the increase in effective population size and overlap among generations.     

No diapause prolongation response of <Emphasis Type="Italic">Daphnia</Emphasis> in the presence of planktivorous mosquitofish (<Emphasis Type="Italic">Gambusia holbrooki</Emphasis>)

Recent research suggests that planktonic organisms in resting stages can perceive predators chemically and delay hatching to evade predation. Using a co-occurring predator–prey pair (Gambusia holbrooki Girard and Daphnia curvirostris Eylmann) from a Mediterranean floodplain wetland, this study tested for such predator-avoidance behaviour of Daphnia. Results show that hatching patterns of Daphnia were not reduced by the presence of different biomass levels of Gambusia. This could be due to the fact that the density of Gambusia in the wetland is high from late spring to late autumn, suggesting that delayed hatchlings would face increased mortality through consumptive predation.     

Behavioural interactions between the introduced plague minnow Gambusia holbrooki and the vulnerable native Australian ornate rainbowfish Rhadinocentrus ornatus,under experimental conditions

The impact of the plague minnow Gambusia holbrooki on the ornate rainbowfish Rhadinocentrus ornatus was investigated by comparing the behavioural response and microhabitat preferences of populations of R. ornatus collected from locations that are sympatric and allopatric with G. holbrooki. Rhadinocentrus ornatus populations from sympatric areas exhibited a significantly higher frequency of intraspecific chases, spent significantly more time at an intermediate depth and were nipped significantly less often by G. holbrooki compared to the allopatric R. ornatus populations. The frequency of intraspecific chases by all R. ornatus populations were greatest immediately following G. holbrooki exposure and increased further with repeated exposure. Activity levels were also significantly higher in the presence of G. holbrooki. Gambusia holbrooki and the allopatric R. ornatus populations showed very similar microhabitat preferences, whereas the preferences for the sympatric R. ornatus populations have shifted to facilitate cohabitation with G. holbrooki. The results suggest that sympatric populations of R. ornatus have evolved or developed behavioural responses to G. holbrooki through niche and character shifts. The implications are discussed in relation to the conservation management of R. ornatus and other threatened species.