Androgen and prolactin manipulation do not induce changes in immunocompetence measures in a fish with male parental care.

Prolactin and 11-ketotestosterone (11-KT) are important reproductive hormones in fishes, which may also influence immunocompetence. The immunocompetence handicap hypothesis states that higher androgen concentrations that support secondary sex traits are traded off against a decrease in immune system function. To test the relationships between these hormones and immunocompetence, we experimentally manipulated 11-ketotestosterone and prolactin in the freshwater fish, bluegill (Lepomis macrochirus) during parental care using implants that contained either 11-KT, prolactin, or an inert control. We vaccinated individuals to stimulate the acquired immune response, then measured immunocompetence as the number of granulocytes, lymphocytes and monocytes, and the expression of interleukin 8 in each sample. We did not observe any significant differences in the immune measures among the hormone treatments. Our results indicate that in bluegill, there is no trade-off between androgens or prolactin and immunocompetence.

Temperature-dependent plasticity mediates heart morphology and thermal performance of cardiac function in juvenile Atlantic salmon (Salmo salar).

In many fishes, upper thermal tolerance is thought to be limited in part by the heart's ability to meet increased oxygen demands during periods of high temperature. Temperature-dependent plasticity within the cardiovascular system may help fish cope with the thermal stress imposed by increasing water temperatures. In this study, we examined plasticity in heart morphology and function in juvenile Atlantic salmon (Salmo salar) reared under control (+0°C) or elevated (+4°C) temperatures. Using non-invasive Doppler echocardiography, we measured the effect of acute warming on maximum heart rate, stroke distance and derived cardiac output. A 4°C increase in average developmental temperature resulted in a >5°C increase in the Arrhenius breakpoint temperature for maximum heart rate and enabled the hearts of these fish to continue beating rhythmically to temperatures approximately 2°C higher than for control fish. However, these differences in thermal performance were not associated with plasticity in maximum cardiovascular capacity, as peak measures of heart rate, stroke distance and derived cardiac output did not differ between temperature treatments. Histological analysis of the heart revealed that while ventricular roundness and relative ventricle size did not differ between treatments, the proportion of compact myocardium in the ventricular wall was significantly greater in fish raised at elevated temperatures. Our findings contribute to the growing understanding of how the thermal environment can affect phenotypes later in life and identify a morphological strategy that may help fishes cope with acute thermal stress.

Effect of isotocin on shoaling behaviour of the Guppy (Poecilia reticulata).

Pro-social effects of oxytocin and its homologues are well-documented in birds and mammals. However, in fishes, the effect of isotocin, the homologue of oxytocin, on social behaviour is less clear. Studies in fishes have generally shown no effect of isotocin on social behaviours or even an anti-social effect. In our study, we measured association preference for conspecifics in 92 adult guppies (46 females and 46 males), half of which were injected with isotocin and the other half with an isotocin antagonist. We found that individuals injected with isotocin spent 29% more time associating with conspecifics than individuals injected with an isotocin antagonist. The effect of isotocin on association time did not differ between males and females. Our study provides some of the first evidence of a pro-social effects of isotocin in a fish and suggests that in fishes, isotocin may have a homologous role to oxytocin, at least in promoting shoaling behaviour.

A test of the effects of androgens on immunity: No relationship between 11-ketotestosterone and immune performance in bluegill (Lepomis macrochirus).

The immunosuppressive effects of androgens are a key component of the immunocompetence handicap hypothesis (ICHH). Here, we use bluegill sunfish (Lepomis macrochirus) to test two predictions arising from this hypothesis: (1) natural circulating concentrations of the androgen 11-ketotestosterone (11-KT) will be negatively related with measures of immunity, and (2) immune stimulation will lower circulating 11-KT concentration. We found no evidence for a relationship between natural circulating 11-KT concentration and measures of immunity (lymphocyte and granulocyte counts, respiratory burst, cytokine mRNA levels), and an immune stimulation with Vibrio vaccine did not affect circulating 11-KT concentration. We also performed a meta-analysis of immune stimulation studies to help interpret our results, and report evidence suggesting that immune stimulation has weaker effects on androgen levels in fishes compared to other vertebrates. These results suggest that the ICHH may not apply to all vertebrates, although it remains premature to state what factors account for the weaker evidence in fishes that androgens are immunosuppressive.

Evolution of kin recognition mechanisms in a fish.

Both selection and phylogenetic history can influence the evolution of phenotypic traits. Here we used recently characterized variation in kin recognition mechanisms among six guppy populations to explore the phylogenetic history of this trait. Guppies can use two different kin recognition mechanisms: either phenotype matching, in which individuals are identified based on comparison with a recognition template, or familiarity, in which individuals are remembered based on previous interactions. Across the six populations, we identified four transitions in recognition mechanism: phenotype matching evolved once and was subsequently lost in a single population, whereas familiarity evolved twice. Based on a molecular clock, these transitions occurred among populations that had diverged on a timescale of hundreds of thousands of years, which is two orders of magnitude faster than previously documented transitions in recognition mechanisms. A randomization test provided no evidence that recognition mechanisms were constrained by phylogeny, suggesting that recognition mechanisms have the capacity to evolve rapidly, although the specific selection pressures that may be contributing to variation in recognition mechanisms across populations remain unknown.

Reproductive success in wild and hatchery male coho salmon.

Salmon produced by hatcheries have lower fitness in the wild than naturally produced salmon, but the factors underlying this difference remain an active area of research. We used genetic parentage analysis of alevins produced by experimentally mixed groups of wild and hatchery coho salmon (Oncorhynchus kisutch) to quantify male paternity in spawning hierarchies. We identify factors influencing paternity and revise previously published behavioural estimates of reproductive success for wild and hatchery males. We observed a strong effect of hierarchy size and hierarchy position on paternity: in two-male hierarchies, the first male sired 63% (±29%; s.d.) of the alevins and the second male 37% (±29%); in three-male hierarchies, the first male sired 64% (±26%), the second male 24% (±20%) and the third male 12% (±10%). As previously documented, hatchery males hold inferior positions in spawning hierarchies, but we also discovered that hatchery males had only 55-84% the paternity of wild males when occupying the same position within a spawning hierarchy. This paternity difference may result from inferior performance of hatchery males during sperm competition, female mate choice for wild males, or differential offspring survival. Regardless of its cause, the combination of inferior hierarchical position and inferior success at a position resulted in hatchery males having only half (51%) the reproductive success of wild males.

Complete genome analysis of a frog virus 3 (FV3) isolate and sequence comparison with isolates of differing levels of virulence.

BACKGROUND: Frog virus 3 (FV3) is the type species of the genus Ranavirus, and in the past few decades, FV3 infections have resulted in considerable morbidity and mortality in a range of wild and cultivated amphibian species in the Americas, Europe, and Asia. The reasons for the pathogenicity of FV3 are not well understood. FINDINGS: We investigated three FV3 isolates designated SSME, wt-FV3, and aza-Cr, and reported that our wt-FV3 and aza-Cr strains showed similar levels of virulence, while SSME was the least virulent in an in vivo study with Lithiobates pipiens tadpoles. Using 454 GS-FLX sequencing technology, we sequenced SSME and compared it to the published wt-FV3 genome. SSME had multiple amino acid deletions in ORFs 49/50L, 65L, 66L, and 87L, which may explain its reduced virulence. We also investigated repeat regions and found that repeat copy number differed between isolates, with only one group of 3 isolates and 1 pair of isolates being identical at all 3 locations. CONCLUSIONS: In this study we have shown that genetic variability is present between closely related FV3 isolates, both in terms of deletions/insertions, and even more so at select repeat locations. These genomic areas with deletions/insertions may represent regions that affect virulence, and therefore require investigation. Furthermore, we have identified repeat regions that may prove useful in future phylogeographical tracking and identification of ranaviral strains across different environmental regions.

Alternative male reproductive tactics drive asymmetrical hybridization between sunfishes (Lepomis spp.).

The potential role of alternative reproductive tactics in circumventing premating isolating mechanisms and driving hybridization between species has long been recognized, but to date there is little empirical support from natural systems. Hybridization occurs between bluegill (Lepomis macrochirus) and pumpkinseed sunfish (Lepomis gibbosus) and it is known to be asymmetrical (male bluegill × female pumpkinseed). Here, we test whether this pattern is driven by a recognition failure by pumpkinseed females or by an alternative cuckolder reproductive tactic in bluegill males. Using genetic parentage data, we found that bluegill cuckolders fathered 24.9% of the larvae in bluegill nests, but no evidence that pumpkinseed females spawned in bluegill nests. Pumpkinseed cuckolders fathered 8.7% of the larvae in pumpkinseed nests, whereas bluegill cuckolders fathered 13.6% of the larvae in those nests. Bluegill cuckolders thus frequently spawn in pumpkinseed nests and are responsible for the asymmetrical hybridization between the species. We discuss the evolutionary consequences of interactions between bluegill and pumpkinseed and the role of alternative reproductive tactics in adaptation and introgression.

A genetic test of sexual size dimorphism in pre-emergent chinook salmon.

Sex differences in early development may play an important role in the expression of sexual size dimorphism at the adult stage. To test whether sexual size dimorphism is present in pre-emergent chinook salmon (Oncorhynchus tshawytscha), alevins were reared at two temperatures (10 °C and 15 °C) and sexed using the OtY1 marker on the Y-chromosome. Linear mixed models were used to test for sex differences in alevin size within families while controlling for the random effects of sire and dam nested within sire. Males and females did not differ in weight at 10 °C but males were heavier than females at 15 °C. Sex accounted for 2% of the within-family variance in weight. In addition, at 15°C, the relationship between weight and sex was greater in families with larger eggs. Whereas male-biased sexual size dimorphism was present at the juvenile stage, female-biased sexual size dimorphism was present at sexual maturity. Males were also younger than females at sexual maturity. A head start on growth by males may underlie their earlier maturation at a smaller size, thus leading to female-biased SSD at the adult stage.

Genetic and ecological assessment of population rehabilitation: walleye in Lake Superior.

Sustainable management of exploited species is an ongoing challenge, particularly where populations have collapsed or been depleted by overharvest and habitat alteration. The walleye (Sander vitreus) population in Lake Superior's Black Bay historically supported more than 90% of the commercial walleye harvest from the entire lake, but collapsed in 1968 and has still not recovered despite long-term closure of the fishery. In an effort to rehabilitate this population, hatchery-origin walleye from exogenous sources were released into Black Bay between 2003 and 2005. We used individual-based analysis of genetic data collected between 2007 and 2010 to examine the contributions of different wild sources and hatchery stocking events to the contemporary walleye population in Black Bay. We found that 75% of the walleye in Black Bay originated from above- and below-barrier native populations in the Black Sturgeon River. The hatchery stocking events differed considerably in their effectiveness: the 2003 release of fry had no measurable contribution, whereas the 2004 and 2005 releases of fingerlings contributed 71% and 45% of the fish in their respective age classes. Hatchery and wild fish were similar in size, but hatchery fish rarely utilized the river habitat where Black Bay walleye historically spawned, and there was little genetic evidence of interbreeding or natural recruitment of stocked fish. Overall, our results suggest that restoring habitat connectivity to facilitate wild recruitment has greater potential than further exogenous stocking to contribute to the recovery of walleye in this system.

Genetic evidence for multiple invasions of the eastern subterranean termite into Canada.

Social insects are among the world's most successful species at invading new environments. Their characteristic division of labor can influence their capacity to colonize new habitats, often with negative ecological or economic impact. The social Hymenoptera (i.e., ants, bees, and wasps), are well studied in this regard, but much less is known about the invasive biology of termites (Isoptera). In this study we use province-wide sampling and a population genetic analysis to infer the minimum number of eastern subterranean termite [Reticulitermes flavipes (Kollar)] introductions into Ontario (Canada). Structure analysis of multilocus microsatellite genotypes grouped the 30 collection points into K = 3 genetic clusters, suggesting as many three independent introductions into southern Ontario. Levels of genetic diversity were higher in termites from the Pelee region than in termites from Toronto and other Ontario cities, suggesting that these Pelee termite populations are potentially older and native to Ontario. A single origin scenario, in which all populations stem from a single source, therefore is not supported by the genetic data. Instead, our analysis suggests multiple independent introductions of this highly social, subterranean termite into Ontario, where the species is now well established as a structural pest of urban habitats.

Sexual conflict inhibits female mate choice for major histocompatibility complex dissimilarity in Chinook salmon.

In many species females prefer major histocompatibility complex (MHC) dissimilar mates, which may improve offspring resistance to pathogens. However, sexual conflict may interfere with female preference when males attempt to mate with all females, regardless of compatibility. Here we used semi-natural spawning channels to examine how mating behaviour and genetic similarity at the MHC class II peptide binding region affected parentage patterns in Chinook salmon (Oncorhynchus tshawytscha). We found that females directed aggression at more MHC-similar males than expected by chance, providing a possible mechanism of female MHC choice in salmon. Males also directed aggression towards MHC-similar females, which was consistent with males harassing unreceptive mates. Males' aggression was positively correlated with their reproductive success, and it appeared to overcome female aversion to mating with MHC-similar males, as females who were the target of high levels of male aggression had lower than expected MHC divergence in their offspring. Indeed, offspring MHC divergence was highest when the sex ratio was female-biased and male harassment was likely to be less intense. These data suggest that male harassment can reduce female effectiveness in selecting MHC-compatible mates, and sexual conflict can thus have an indirect cost to females.