BALANCING SELECTION IN A DESERT STREAM-DWELLING FISH,POECILIOPSIS MONACHA

The desert stream-dwelling fish Poeciliopsis monacha is exposed to extreme seasonal and spatial variation in physical stresses. We examined four diallelic gene loci (Ldh-1, Idh-2, Pgd, and Ck-A) in P. monacha and tested whether genotypes were associated with differential survival of individuals exposed to acute stress. For each locus, the common allele was associated with higher survival during heat and hypoxic stress, whereas the alternate allele was associated with higher survival during cold stress. In most cases, survival of heterozygotes was intermediate and they exhibited less variance in survival than corresponding homozygotes. Identification of substantial linkage disequilibrium in these fish confounds our ability to discern whether the allozymes are the direct targets of selection, or if they just mark chromosomal regions that contain the true modifiers of survival. Nevertheless, the present results clearly identified balancing processes that can serve to stabilize genetic polymorphism in this species.     

Heat induced liver cell proliferation in the livebearing fish Poeciliopsis

Synopsis Liver regeneration is induced by heat stress in the small viviparous fish, Poeciliopsis. Acute exposure to sublethal temperatures, one to two degrees below their killing temperature, damages tissue and initiates liver cell proliferation in P. lucida, P. monacha, and P. monacha-lucida hybrid clones, SYN-4 and SYN-5. Regeneration of liver cells began within 1–2 days following heat stress and proceeded over 5 days. Peak cell proliferation occurred 2–3 days after treatment in fish of all four genotypes. Cell proliferation was induced in the two all-female clones, SYN-4 and SYN-5, by exposure to 40.5° C for 60 minutes. This treatment imposed mortalities of 17.9% and 16.7%, respectively, whereas reduction of the temperature to 39.5°C and reduction of the time to 30 minutes resulted in no mortalities without significantly lowering the level of cell proliferation (p > 0.05). Liver cell proliferation induced by both heat treatments was significantly higher (p < 0.05) in the SYN-5 hybrids than in SYN-4. The induction of liver cell proliferation with sublethal temperature exposures is discussed as it may relate to chemical carcinogenesis in both feral and laboratory fish. Acute heat exposure may be used experimentally in fish as an independent stimulus for liver cell proliferation in carcinogenesis studies. In poikilothermic animals-heat exposure offers an alternative to surgical removal of approximately two-thirds of the liver, the method most frequently used in rodents to study the process of liver regeneration.     

Measuring Ucrit and endurance: equipment choice influences estimates of fish swimming performance

This study compared the critical swimming speed (U_crit) and endurance performance of three Australian freshwater fish species in different swim‐test apparatus. Estimates of U_crit measured in a large recirculating flume were greater for all species compared with estimates from a smaller model of the same recirculating flume. Large differences were also observed for estimates of endurance swimming performance between these recirculating flumes and a free‐surface swim tunnel. Differences in estimates of performance may be attributable to variation in flow conditions within different types of swim chambers. Variation in estimates of swimming performance between different types of flumes complicates the application of laboratory‐based measures to the design of fish passage infrastructure.     

Linkage conservation of homologous esterase loci in fish (Cyprinodontoidei: Poeciliidae)

Homologies among esterase isozymes in fish in the poeciliid genera Poeciliopsis and Xiphophorus are proposed. Esterase homologies are based on their tissue distributions and inhibition and substrate properties. The five esterases include two carboxylesterases, one eserine sulfate-sensitive esterase, and two esterases resistant to inhibition, one of which reacts only with acetate esters. Linkage studies in Poeciliopsis monacha indicate that the loci encoding the two carboxylesterases are linked to each other and to the locus for eye-specific lactate dehydrogenase. Comparisons of the linkage reported here with earlier studies in Xiphophorus suggest that there is a large region of linkage homology in the genetic maps of Poeciliopsis and Xiphophorus.This work was supported by grants from the National Science Foundation (DEB76-19285) to R. C. Vrijenhoek and the Charles and Johanna Busch Fund to R. C. Vrijenhoek and N. H. Hart. J. F. L. and P. J. P. were supported by U.S. Public Health Service Genetics Training Grant GM07129-04.     

Displacement,velocity preference,and substrate use of three native California stream fishes in simulated pulsed flows

We studied whether juvenile fishes were able to maintain swimming speed and position during simulated river pulsed flows in a laboratory flume. We used a glass flume (15.24 × 0.6 m) with river-rock substrate to determine the longitudinal displacement, movement distances and frequencies, velocity selection, and substrate use of juvenile (SL range: 6.1 ± 0.2 cm) hardhead Mylopharodon conocephalus (n = 13), rainbow trout Oncorhynchus mykiss (n = 11), and Sacramento sucker Catostomus occidentalis (n = 12) during a 100-min flow pulse, as velocity changed from slow to medium, fast, medium, and slow. Fish were capable of maintaining swimming speed and position up to the maximum flume velocity of 0.46 m·s⁻¹, except for one hardhead that impinged on the rear fish screen. Fish swam faster in the flume during the medium and fast intervals than the slow intervals, but fish speeds were similar among the medium and faster intervals, when some fish took cover behind the rock substrate. In comparison with a Brett-type swim-tunnel, fish showed less increase in mean swimming speed as the flume velocity increased. Fish in the flume were able to use the rock substrate as hydraulic cover, decreasing the encountered water velocity, and, presumably, conserving energy.     

The influence of flume length and group size on swimming performance in shortnose sturgeon Acipenser brevirostrum

The main objectives of this study were to determine optimal methodologies to assess the general swimming performance of juvenile shortnose sturgeon Acipenser brevirostrum. Swimming densities (group v. individual swimming) and flume length (2 v. 1 m) were altered to verify if any of those variables affected performance (i.e. time to fatigue) during critical swimming (U(crit)) and endurance tests. Results for both U(crit) and endurance swimming were not significantly different between fish swum in groups of five or fish swum individually. The U(crit) values, however, were c. 22% higher for fish swum in a longer flume. Although swimming fish in groups did not improve swimming performance, group swimming lowered the variance of the data. Results also reveal that juvenile A. brevirostrum may not possess an ability to swim at high speeds (i.e. burst phase) for long periods.     

Effect of thermo-velocity barriers on fish: influence of water temperature,flow velocity and body size on the volitional swimming capacity of northern straight-mouth nase (Pseudochondrostoma duriense)

Water temperature and flow velocity directly affect the fish swimming capacity, and thus, both variables influence the fish passage through river barriers. Nonetheless, their effects are usually disregarded in fishway engineering and management. This study aims to evaluate the volitional swimming capacity of the northern straight-mouth nase (Pseudochondrostoma duriense), considering the possible effects of water temperature, flow velocity and body size. For this, the maximum distance, swim speed and fatigue time (FT) were studied in an outdoor open-channel flume in the Duero River (Burgos, Spain) against three nominal velocities (1.5, 2.5 and 3 m s⁻¹) and temperatures (5.5, 13.5 and 18.5°C), also including the changes between swimming modes (prolonged and sprint). Results showed that a nase of 20.8 cm mean fork length can develop a median swim speed that exceeds 20.7 BL s⁻¹ (4.31 m s⁻¹) during a median time of 3.4 s in sprint mode, or 12.2 BL s⁻¹ (2.55 m s⁻¹) for 23.7 s in prolonged mode under the warmest scenario. During prolonged swimming mode, fish were able to reach further distances in warmer water conditions for all situations, due to a greater swimming speed and FT, whereas during sprint mode, warmer conditions increased the swim speed maintaining the FT. In conclusion, the studied temperature range and flow velocity range influence fish swimming performance, endurance and distance travelled, although with some differences depending on the swimming mode. The provided information goes a step forward in the definition of real fish swimming capacities, and in turn, will contribute to establish clear passage criteria for thermo-velocity barriers, allowing the calculation of the proportion of fish able to pass a barrier under different working scenarios, as well designing of the optimized solutions to improve the fish passage through river barriers.     

The effect of substratum type on aspects of swimming performance and behaviour in shortnose sturgeon Acipenser brevirostrum

The swimming performance and associated swimming behaviour (i.e. substratum‐skimming, station‐holding and free swimming) were assessed in shortnose sturgeon Acipenser brevirostrum during critical swimming and endurance swimming tests over a rough and a smooth substratum. It was hypothesized that the addition of a rough substratum in the swimming flume may provide a surface for the A. brevirostrum to grip and offer an energetic advantage. Substratum type did not affect the critical swimming performance, but A. brevirostrum consistently performed more bottom behaviours (i.e. substratum‐skimming and station‐holding) while on a smooth substratum. Acipenser brevirostrum had little contact with the rough substratum until the velocity was >1 body length s^?1. Endurance swimming time was significantly lower for A. brevirostrum over the rough bottom at the highest velocity (30 cm s^?1) which may be attributed to the observed increase in free swimming and decrease in bottom behaviours. During endurance swimming, the rough substratum was mainly used at intermediate velocities, suggesting that there may be a stability cost associated with being in contact with the rough substratum at certain velocities.     

Quantitative evidence that both Hsc70 and Hsp70 contribute to thermal adaptation in hybrids of the livebearing fishes Poeciliopsis

The 70-kilodalton heat shock protein family is composed of both environmentally inducible (Hsp) and constitutively expressed (Hsc) family members. While the role of the constitutively expressed stress proteins in thermotolerance is largely unknown, de novo expression stress proteins in response to elevated temperatures has been associated with increased thermotolerance in many cell lines, developing embryos and adult organisms. Distinct, hemiclonal hybrids between the livebearing fish species Poeciliopsis monacha and P. lucida varied in their abilities to survive temperature stress, with survival being greatest when rates of temperature increase to 40°C were slowest and when P. monacha genomes were combined with a sympatric P. lucida genome. Quantification of Hsp70 under heat shock conditions and Hsc70 under normal physiological conditions indicated that variation in survival among hemiclones was best explained by the combined effects of these two proteins. Similar complex interactions between maternal and paternal genomes and rate of temperature increase were found to underline patterns of survival, Hsp70 accumulation and Hsc70 abundance. These data suggest that the relationship between Hsps and thermotolerance is more intricate than previously thought and that Hsps contribute to thermal adaptation in these fishes through genetic interactions specific to particular environments.     

Elevated incubation temperature improves later-life swimming endurance in juvenile Chinook salmon,Oncorhynchus tshawytscha

The effect of incubation and rearing temperature on muscle development and swimming endurance under a high-intensity swimming test was investigated in juvenile Chinook salmon (Oncorhynchus tshawytscha) in a hatchery experiment. After controlling for the effects of fork length (L_F) and parental identity, times to fatigue of fish were higher when fish were incubated or reared at warmer temperatures. Significant differences among combinations of pre- and post-emergence temperatures conformed to 15–15°C > 15–9°C > 9–9°C > 7–9°C > 7–7°C in 2011 when swimming tests were conducted at 300 accumulated temperature units post-emergence and 15–9°C > (7–9°C = 7–7°C) in 2012 when swimming tests were conducted at an L_F of c. 40 mm. The combination of pre- and post-emergence temperatures also affected the number and size of muscle fibres, with differences among temperature treatments in mean fibre cross-sectional area persisting after controlling for L_F and parental effects. Nonetheless, neither fibre number nor fibre size accounted for significant variation in swimming endurance. Thus, thermal carryover effects on swimming endurance were not mediated by thermal imprinting of muscle structure. This is the first study to test how temperature, body size and muscle structure interact to affect swimming endurance during early development in salmon.     

Fish swimming stride by stride: speed limits and endurance

Summary Steadily swimming fish show a species-specific stride length and tail tip amplitude. These are constant over the entire speed range if expressed as a fraction of the body length. The speed of a fish equals the stride length times the tail beat frequency. We describe how maximum tail beat frequencies, and hence maximum swimming speeds, are related to temperature and body length.Maximum sustained swimming speeds, endurance during swimming at higher speeds, and maximum burst velocities of 27 species are compared. The rate of decline of endurance with increasing speed is either gradual or steep, with only a few cases in between Steady swimmers show the steepest decline.The published effects of temperature on endurance are not consistent.The effect of body size on the endurance curve could be investigated for two species. The maximum sustained speed decreases with increasing length, and the slope of the endurance curves steepens with increasing length with the same factor in both species. The maximum burst speed is 10 Ls^-1 on average.     

FLAT REACTION NORMS AND “FROZEN” PHENOTYPIC VARIATION IN CLONAL SNAILS (POTAMOPYRGUS ANTIPODARUM)

The Frozen Niche-Variation hypothesis (FNV) suggests that clones randomly sample and “freeze” the genotypes of their ancestral sexual populations. Hence, each clone expresses only a fraction of the total niche-use variation observed in the sexual population, which may lead to selection for ecological specialization and coexistence of clones. A generalized form of the FNV model suggests that the same is true for life-history (as well as other) traits that have important fitness consequences, but do not relate directly to niche use. We refer to the general form of the model as the Frozen Phenotypic Variation (FPV) model. A mixed population of sexual and parthenogenetic snails (Potamopyrgus antipodarum) in a New Zealand lake allowed us to examine the phenotypic variation expressed by coexisting clones in two benthic habitats, and to compare that variation to the sexual population. Three clones were found primarily in an aquatic macrophyte zone composed of Isoetes kirkii (1.5–3.0 m deep), and three additional clones were found in a deeper macrophyte zone composed of Elodea canadensis (4.0–6.0 m deep). These clones showed significant variation between habitats, which mirrored that observed in the sexual population. Specifically, clones and sexuals from the deeper habitat matured at a larger size and had larger broods. There was also significant among-clone variation within habitats; and as expected under the FPV model, the within-clone coefficients of variation for size at maturity were low in both habitats when compared to the sexual population. In addition, we found four clones that were common in both macrophyte zones. The reaction norms of these clones were flat across habitats, suggesting little phenotypic plasticity for morphology or life-history traits. Flat reaction norms, high among-clone variation, and low coefficients of variation (relative to the sexual population) are in accordance with the FPV model for the origin of clonal lineages. We also measured the prevalence of infection by trematode larvae to determine whether clones are inherently more or less infectable, or whether they are freezing phenotypic variation for resistance from the sexual population. We did this in the deep habitats of the lake where recycling of the parasite by the vertebrate host is unlikely, thereby reducing the complications raised by frequency-dependent responses of parasites to host genotypes. We found no indication that clones are either more or less infectable than the resident sexual population. Taken together, our results suggest that phenotypic variation for both life-history traits and resistance to parasites is frozen by clones from the local sexual population.     

Swimming performance and invasion potential of the round goby

European round gobies (Neogobius melanostomus) are displacing several important native North American fish species. Controlling their invasion is contingent on understanding their swimming inclination and potential. We assessed goby swimming inclination by recording activity in a 2 m flume over a ~24 h period, and swimming potential using a critical swimming (U _crit) test, as well as burst tests in still and flowing water. When given the choice to move, gobies covered as much as 14 m/h, with a slight bias towards nocturnal activity and an overall upstream preference. When confined and coerced to perform a U _crit test, they burst-and-held to achieve 35.5 ± 1.1 cm/s. Thirty minutes following U _crit, they were able to burst-and-coast in a sprint test to almost twice this speed. In still water, they exhibited startle bursts of up to 163 cm/s. We provide a swimming endurance model that indicates flow rates would need to be >125 cm/s to prevent upstream movement, and free of refuge areas in which to recover. The current study shows that the round goby is a surprisingly powerful swimmer with the capacity to continue its invasion should hydrologic control be absent.     

The swimming endurance of threespine sticklebacks, Gasterosteus aculeatus L., from the Afon Rheidol, Wales

The endurance of threespine sticklebacks, Gasterosteus aculeatus , swimming with pectoral fin locomotion at 20° C in a laboratory flume was measured. Each trial lasted a maximum of 480 min. At a speed of 4 body lengths per sec (L s⁻¹) all fish were still swimming at the end of the trial, but endurance decreased at higher speeds. At speeds of 5 or 6 L s⁻¹ (20–30 cm s⁻¹) a few fish still maintained labriform locomotion for the 480 min. However, at a speed of 7 L s⁻¹ all fish furled their pectoral fins and used body and caudal fin propulsion but fatigued rapidly. During sustained swimming, fish could cover distances of 6 km or more. No significant differences between males and females were found.     

Trade-off between steady and unsteady swimming underlies predator-driven divergence in Gambusia affinis

Differences in predation intensity experienced by organisms can lead to divergent natural selection, driving evolutionary change. Western mosquitofish (Gambusia affinis) exhibit larger caudal regions and higher burst-swimming capabilities when coexisting with higher densities of predatory fish. It is hypothesized that a trade-off between steady (constant-speed cruising; important for acquiring resources) and unsteady (rapid bursts and turns; important for escaping predators) locomotion, combined with divergent selection on locomotor performance (favouring steady swimming in high-competition scenarios of low-predation environments, but unsteady swimming in high-predation localities) has caused such phenotypic divergence. Here, I found that morphological differences had a strong genetic basis, and low-predation fish required less hydromechanical power during steady swimming, leading to increased endurance. I further found individual-level support for cause-and-effect relationships between morphology, swimming kinematics and endurance. Results indicate that mosquitofish populations inhabiting low-predation environments have evolved increased steady-swimming abilities via stiffer bodies, larger anterior body/head regions, smaller caudal regions and greater three-dimensional streamlining.     

Facing warm temperatures during migration: cardiac mRNA responses of two adult Oncorhynchus nerka populations to warming and swimming challenges

The main findings of the current study were that exposing adult sockeye salmon Onchorhynchus nerka to a warm temperature that they regularly encounter during their river migration induced a heat shock response at an mRNA level, and this response was exacerbated with forced swimming. Similar to the heat shock response, increased immune defence‐related responses were also observed after warm temperature treatment and with a swimming challenge in two different populations (Chilko and Nechako), but with some important differences. Microarray analyses revealed that 347 genes were differentially expressed between the cold (12–13° C) and warm (18–19° C) treated fish, with stress response (GO:0006950) and response to fungus (GO:0009620) elevated with warm treatment, while expression for genes involved in oxidative phosphorylation (GO:0006119) and electron transport chain (GO:0022900) elevated for cold‐treated fish. Analysis of single genes with real‐time quantitative PCR revealed that temperature had the most significant effect on mRNA expression levels, with swimming and population having secondary influences. Warm temperature treatment for the Chilko population induced expression of heat shock protein (hsp) 90α, hsp90β and hsp30 as well as interferon‐inducible protein. The Nechako population, which is known to have a narrower thermal tolerance window than the Chilko population, showed even more pronounced stress responses to the warm treatment and there was significant interaction between population and temperature treatment for hsp90β expression. Moreover, significant interactions were noted between temperature treatment and swimming challenge for hsp90α and hsp30, and while swimming challenge alone increased expression of these hsps, the expression levels were significantly elevated in warm‐treated fish swum to exhaustion. In conclusion, it seems that adult O. nerka currently encounter conditions that induce several cellular defence mechanisms during their once‐in‐the‐lifetime migration. As river temperatures continue to increase, it remains to be seen whether or not these cellular defences provide sufficient protection for all O. nerka populations.     

Exposure to a common urban pollutant affects the survival and swimming behaviour of creek chub (Semotilus atromaculatus)

Anthropogenic effects on the aquatic environment are ever present and ever increasing and while a plethora of aquatic contaminants are known to affect fishes, one ubiquitous and increasingly prevalent world-wide urban runoff pollutant is frequently disregarded, and that is pet waste. While dog waste has been identified as a significant factor contributing to bacteria and nutrient loading within receiving waters and the associated water quality changes are known to affect fishes, the impact of uncollected dog faeces on urban fish populations has never been directly investigated. In this study we exposed creek chub (Semotilus atromaculatus), a widespread tolerant stream minnow, to various realistic concentrations of dog waste as simulated urban park runoff testing both fresh and dried dog faeces in both stagnant and aerated water for 96 h to investigate the impact on fish survival and behaviour. Creek chub percentage mortality increased significantly relative to controls and across an exposure gradient and was likely caused by anoxic conditions. Survivors were initially smaller while those that died were initially larger and presented with abnormal abdominal subdermal lesions post-exposure. Additional indicators of physiological stress included significantly increased rates of aquatic surface respiration and changes in flume test derived swimming motivation metrics with increased exposure concentrations. Both mortality and behavioural responses were alleviated by aeration. Furthermore, trials with fresh and dried faeces differed only in time-to-death and swimming metrics where results from dried trials were similar to those from aerated experiments. Results demonstrated the impact that the global dog waste management problem can have on aquatic communities with effects on creek chub likely to be more severe for less pollution-tolerant species and also likely to be exacerbated under future scenarios that consider climate change and increased urbanization.     

Repeat UCrit and endurance swimming in juvenile shortnose sturgeon (Acipenser brevirostrum)

Previous results show that juvenile shortnose sturgeon are steady swimmers and, compared with salmonids, generally have low critical swimming (UCrit) and endurance swimming capacities. Most studies on swimming capacities of sturgeon, and other fishes, include those where fish have only been swum once and the metrics of swimming performance are assessed (e.g., time swum, speed achieved). Under natural conditions, there are ample instances where fish undergo multiple swimming cycles when traversing fish ways, culverts and other sources of fast water flow. While some evidence exists for salmonids, the effects of repeat swimming are not well known for sturgeon. The current study consisted of two experiments. The first examined the UCrit of juvenile shortnose sturgeon following three consecutive swimming trials with a 30 min recovery period between subsequent tests. The second examined the endurance swimming capacities of juvenile shortnose sturgeon following three consecutive swimming trials with a 60 min recovery period between subsequent tests. Our findings indicate that (i) UCrit was consistent (~2 body lengths/s) among swimming trials; (ii) significant individual variation exists between individuals in the endurance swimming trials; and (iii) consistent results exist for individuals across swimming trials in both the UCrit and the endurance swimming tests. These results suggest that juvenile shortnose sturgeon have a high recovery capacity, and their behaviour and morphology likely reflect aspects of their swimming capacities.     

Comparative phylogeography of livebearing fishes in the genera Poeciliopsis and Poecilia (Poeciliidae: Cyprinodontiformes) in central Mexico

Aim To test the hypothesis that the vicariant event responsible for north–south divergences in two clades of the fish genus Poeciliopsis Regan was also responsible for north–south divergences in the fish Poecilia butleri Jordan. Location Central Mexico. Methods Parsimony, distance, maximum likelihood and Bayesian phylogenetic analyses of a mitochondrial gene. Molecular clock test and Bayesian analyses of divergence time. Results We report concordant phylogeographical patterns between two clades in the genus Poeciliopsis (i.e. the other formed by P. latidens Garman and P. fasciata Meek, and the other formed by P. presidionis Jordan and P. turneri Miller) and the clade of Poecilia butleri, with northern and southern individuals within each clade grouping into separate lineages. There is also evidence for slower substitution rates in Poecilia compared with Poeciliopsis. After taking into account these substitution rate discrepancies with Bayesian relaxed molecular clock analyses, north–south divergences in Poecilia butleri were equivalent to those reported for Poeciliopsis latidens‐fasciata and P. presidionis‐turneri. Main conclusions The same Plio‐Pleistocene vicariant event associated with geological activity of the Trans‐Mexican Volcanic Belt (TMVB) appears to have caused divergence in three different freshwater fish lineages. This study is an example of how comparative phylogeography can strengthen inferences about vicariant events in regions of high biological diversity and complex geological history such as the TMVB.     

Assignment of six enzyme loci to multipoint linkage groups in Fishes of the genusPoeciliopsis (Poeciliidae): Designation of linkage groups III–V

Three new linkage groups of enzyme loci are described usingPoeciliopsis monacha × P. viriosa-derived interspecific backcross hybrids. Comparison to known linkage groups of the confamilial genusXiphophorus shows homology betweenXiphophorus linkage group I andPoeciliopsis linkage group III,Xiphophorus linkage group II andPoeciliopsis linkage group I, andXiphophorus linkage group IV andPoeciliopsis linkage group IV. Comparison of the gene content of other fish, amphibians, and mammal syntenic groups suggests retention of plesiomorphic vertebrate gene arrangements in at least two poeciliid linkage groups. Expansion of thePoeciliopsis gene map should be of utility in the identification of tumor regulatory genes through demonstration of linkage to biochemical markers.This work was supported by NSF Grants BSR 19355 and BSR 16569 and NIH Grants CA 44303 and CA 39729 to R. S. Nairn and D.C.M. and NIEHS Grant EHS 1P50ES 0384801A1 to R.J.S.