Variation at enzyme coding loci and correlates of fitness in rainbow trout: a cohort analysis

An association between heterozygosity at six to nine enzyme coding loci and correlates of fitness (survival, age at first maturation) was tested in two cohorts of rainbow trout Oncorhynchus mykiss reared in the laboratory. A subsample offish from the first cohort was reared together for 3 years. There, fish sampled as embryos, juveniles and adults did not differ significantly in multilocus heterozygosity (MLH, numbers of heterozygous loci per fish). However, fish that died during a period of high natural mortality had significantly greater MLH than those sampled before as hatched embryos and later as adults. The number of homozygotes and heterozygotes at three loci differed significantly among different aged fish but in different directions. Allele counts at two of these loci also differed among samples. In a second cohort, the heterozygosity of fish that died during the period of high natural mortality was compared with that of healthy fish collected at the same time; no significant differences in MLH or allele frequencies were detected. Taken together, the lower MLH of the natural mortalities as well as single locus effects observed in the first cohort might be explained by the differential survival of fish with particular genotypes leading to changes in heterozygosity of the entire cohort over time rather than heterozygosity per se . Age of first maturation and heterozygosity (MLH and single locus) were not associated in males but were marginally so in females. Time of ovulation within the spawning season was not associated with the MLH of females.     

Genetic studies of laboratory reared mussels, Mytilus edulis: heterozygote deficiencies, heterozygosity and growth

Offspring derived from several single pair matings and a mass mating of Mytilus edulis were cultured up to the juvenile stage in the laboratory. Samples were collected from the cultures at an early post- settlement ('spat') stage and again at the juvenile stage and were scored at up to nine enzyme loci using cellulose acetate and starch gel electrophoresis. An overall significant deficiency of heterozygotes was evident at the juvenile stage but not at the spat stage. Selection against heterozygotes (underdominance) is proposed as the cause.
For each mating, shell lengths of juvenile mussels were measured and individuals were classed according to the number of loci at which they were heterozygous. No significant correlations were evident between heterozygosity class and either growth (shell length), or coefficient of variation of shell length. It is argued that these results provide support for the 'associative overdominance' rather than the 'overdominance' hypothesis as an explanation for heterozygosity/growth correlations in marine bivalves.
These data also show that although heterozygosity and growth co-occur in most data sets from natural populations, they may have distinctly differing causes.     

Effect of allozyme heterozygosity on basal and induced levels of heat shock protein (Hsp70), in juvenile Concholepas concholepas (Mollusca)

Organisms cope physiologically with extreme temperature by producing heat shock proteins (HSPs). Expression of Hsp70 enhances thermal tolerance and represents a key strategy for ectotherms to tolerate elevated temperature in nature. Synthesis of these proteins, together with other physiological responses to elevated temperatures, increases energy demands. A positive association between multiple and single locus heterozygosity (MLH and SLH, respectively) and individual fitness has been widely demonstrated. In molluscs, MLH can decrease routine metabolic rates and improve energetic status. Juvenile Concholepas concholepas live in the intertidal zone and are constantly exposed to temperature fluctuations. Thus, these young individuals are exposed both to thermal risks and the large metabolic costs required to cope with thermal stress. We evaluated the effects of allozyme MLH and SLH on basal (control animals) and induced (stressed animals) levels of the Hsp70 in juveniles C. concholepas. Juveniles (n = 400) were acclimated at 16 °C for 2 weeks; then 100 animals were exposed to 24 °C (stress) and 100 were kept at 16 °C (control) for 2 and 7 days. The variability of 20 loci was analyzed by starch gel electrophoresis. For SLH effects we used 7 polymorphic loci. We quantified expression of Hsp70 by Western blot analyses. Hsp70 expression increased markedly (~ 90%) with temperature. We found a positive association between MLH and basal and induced levels of Hsp70 in the 2-day exposure experiment. Regardless of temperature, Hsp70 levels increased with MLH (r² = 0.7 and 0.9, for basal and induced levels, respectively) reaching maximal levels in juveniles with intermediate and high MLH levels (2 and 3 loci), and decreasing slightly (but not significantly) in juveniles with highest MLH (≥ 4 heterozygous loci). However, after 7 days of exposure to thermal stress, less heterozygous juveniles attained the same levels of Hsp70 than more heterozygous juveniles. Given the faster increment of Hsp70 in C. concholepas juveniles with intermediate-high levels of MLH, these individuals could be less affected by thermal stress in the intertidal zone. We found an association between specific loci genotype and higher Hsp70 levels (basal or induced). In comparison to homozygous juveniles, heterozygous juveniles for several loci showed higher Hsp70. However, these associations were not for the same loci in juveniles exposed to high temperature for 2 and 7 days. This suggests genotypic variation at some allozyme loci could be more important in the period of initial response to high temperature and others can be more important in the response to the chronic temperature stress.     

Growth rate correlates to individual heterozygosity in the European eel, Anguilla anguilla L

Heterozygosity-fitness correlations (HFCs) have been reported in populations of many species. We provide evidence for a positive correlation between genetic variability and growth rate at 12 allozyme loci in a catadromous marine fish species, the European eel (Anguilla anguilla L.). More heterozygous individuals show a significantly higher length and weight increase and an above average condition index in comparison with more homozygous individuals. To a lesser extent, six microsatellite loci show a similar pattern, with positive but not significant correlations between heterozygosity and growth rate. The HFCs observed could be explained by an effect of either direct allozyme over-dominance or associative overdominance. Selection affecting some of the allozyme loci would explain the greater strength of the HFCs found at allozymes in comparison with microsatellites and the lack of correlation between MLH at allozymes and MLH at microsatellites. Associative overdominance (where allozyme loci are merely acting as neutral markers of closely linked fitness loci) might provide an explanation for the HFCs if we consider that allozyme loci have a higher chance than microsatellites to be in linkage disequilibrium with fitness loci.     

HETEROZYGOSITY‐FITNESS CORRELATIONS IN RAINBOW TROUT: EFFECTS OF ALLOZYME LOCI OR ASSOCIATIVE OVERDOMINANCE?

Previous studies with rainbow trout (Oncorhynchus mykiss) have shown that increased heterozygosity at allozyme loci is correlated with several phenotypic traits associated with fitness. We expected to find a similar effect of heterozygosity at other nuclear loci if these associations are due to loci in linkage disequilibrium with the allozyme loci (i.e., associative overdominance), rather than the allozymes themselves. We examined the association between multiple locus heterozygosity and condition factor at 10 allozyme and 10 microsatellite loci. Individuals that were more heterozygous at allozyme loci had significantly greater condition factor in two hatchery cohorts of rainbow trout (1996 P = 0.006; 1997 P < 0.001). In contrast, there was no evidence at microsatellite loci that increased heterozygosity was associated with greater condition factor. Our results suggest that the observed relationship between heterozygosity at allozyme loci and condition factor in rainbow trout appears to be due to the allozyme loci themselves, rather than associative overdominance. We cannot, however, rule out that differences in the mutation process between allozymes and microsatellites may be responsible for these observations. Regardless of the underlying mechanism, these results support the view that allozymes and microsatellites are differentially affected by natural selection.     

Allozyme Diversity among Different Size Cohorts of Spat of Mytilus trossulus (Bivalvia,Mytilidae) Dwelling together in Amursky Bay,Sea of Japan

Allozyme variability in 16 polymorphic loci in two size cohorts of large (30.0 ± 6.3 mm) and small (9.8 ± 2.3 mm) spat of the mussel Mytilus trossulus (Gould) coinhabiting the intertidal zone in Brazhnikov Bay (Amursky Bay, Sea of Japan) was studied 1988–1989. It was determined that the size cohorts compared were highly similar genetically; Nei's minimum distances were 0.014 and 0.016 in 1988 and in 1989. However, statistically significant differences in allele frequencies were found at 2 loci, both in 1988 and in 1989. The loci differing by allele frequencies in 1988 did not coincide with those in 1989. Significant differences by heterozygosity between the cohorts were found only at the Idg-1 and Est-3 loci, 1988; and at the Aat-1, 6-Fgd, Est-3 loci, 1989; but the average heterozygosity was not significantly different in the cohorts for both years. A deficiency of heterozygotes (Dg) was observed almost at all loci in both cohorts. The mean Dg values were significantly higher in the cohort of small size individuals. Since the environmental conditions for both cohorts were similar, the allozyme differences between them were not a result of natural selection. A possible cause of the allozyme and size differences among the mussel cohorts is the gap in the settling time of larvae, which entails the formation of two size cohorts of mussels. Such a gap was caused by protraction of spawning period of M. trossulus and by the formation of segregate larval pools from various mussel populations differing in allele frequencies.     

Enzyme heterozygosity and growth of rainbow trout reared at two rations

The relationship between heterozygosity at nine enzyme loci and both size and growth in rainbow trout ( Salmo gairdneri ) reared at either low (LRT) or high (HRT) ration for 84 days is reported. The experimental fish were progeny produced from a pooled mating of 25 female and 25 male rainbow trout captured in spawning condition at the Ganaraska River, Ontario. There is a significant negative regression between the number of heterozygous loci per fish and size (fork length and weight) among fish at the beginning of the experiment. Heterozygotes at two loci, Sod and Mdh3,4 , were significantly smaller than homozygotes. A significant negative regression between multilocus heterozygosity and size was detectable among the fish reared at HRT for 84 days but not at LRT. Ration appeared to affect the strength of the association between heterozygosity and growth because at HRT homozygotes at the majority of loci grew faster than heterozygotes, but the reverse was true at LRT.     

META-ANALYSES OF THE ASSOCIATION BETWEEN MULTILOCUS HETEROZYGOSITY AND FITNESS

Meta-analyses of published correlation coefficients between multilocus heterozygosity (MLH) and two fitness surrogates, growth rate and fluctuating asymmetry, suggested that the strength of these correlations are generally weak. A variety of plants and animals was included in the meta-analyses. A statistically homogeneous group of MLH–growth rate correlation coefficients that included both plants and animals yielded a common correlation of r_z = 0.133. A common correlation of r_z = –0.170 was estimated for correlations between MLH and fluctuating asymmetry in three species of salmonid fishes. These results suggest that selection, including overdominance, has at most a weak effect at allozyme loci and cast some doubt on the widely held notion that heterozygosity and individual fitness are strongly correlated.     

Allozyme and RFLP Heterozygosities as Correlates of Growth Rate in the Scallop Placopecten Magellanicus: A Test of the Associative Overdominance Hypothesis

Several studies have reported positive correlations between the degree of enzyme heterozygosity and fitness-related traits. Notable among these are the correlations between heterozygosity and growth rate in marine bivalves. Whether the correlation is the result of intrinsic functional differences between enzyme variants at the electrophoretic loci scored or arises from non-random genotypic associations between these loci and others segregating for deleterious recessive genes (the associative overdominance hypothesis) is a matter of continuing debate. A prediction of the associative overdominance hypothesis, not shared by explanations that treat the enzyme loci as causative agents of the correlation, is that the correlation is not specific to the type of genetic marker used. We have tested this prediction by scoring heterozygosity at single locus nuclear restriction fragment length polymorphisms (RFLPs) in a cohort of juvenile scallops (Placopecten magellanicus) in which growth rate was known to be positively correlated with an individual's degree of allozyme heterozygosity. A total of 222 individuals were scored for their genotypes at seven allozyme loci, two nonspecific protein loci of unknown function and eight nuclear RFLPs detected by anonymous cDNA probes. In contrast to the enzyme loci, no correlation was observed between growth rate and the degree of heterozygosity at the DNA markers. Furthermore, there was no relationship between the magnitude of heterozygote deficiency at a locus and its effect on the correlation. The differences observed between the effects of allozyme and RFLP heterozygosity on growth rate provide evidence against the associative overdominance hypothesis, but a strong case against this explanation must await corroboration from similar studies in different species.     

Juvenile socio-sexual experience determines lifetime sperm expenditure and adult survival in a polygamous moth,Ephestia kuehniella

Male animals often adjust their sperm investment in response to sperm competition environment. To date, only a few studies have investigated how juvenile sociosexual settings affect sperm production before adulthood and sperm allocation during the first mating. Yet, it is unclear whether juvenile sociosexual experience (1) determines lifetime sperm production and allocation in any animal species; (2) alters the eupyrene : apyrene sperm ratio in lifetime ejaculates of any lepidopteran insects, and (3) influences lifetime ejaculation patterns, number of matings and adult longevity. Here we used a polygamous moth, Ephestia kuehniella, to address these questions. Upon male adult emergence from juveniles reared at different density and sex ratio, we paired each male with a virgin female daily until his death. We dissected each mated female to count the sperm transferred and recorded male longevity and lifetime number of matings. We demonstrate for the first time that males ejaculated significantly more eupyrenes and apyrenes in their lifetime after their young were exposed to juvenile rivals. Adult moths continued to produce eupyrene sperm, contradicting the previous predictions for lepidopterans. The eupyrene : apyrene ratio in the lifetime ejaculates remained unchanged in all treatments, suggesting that the sperm ratio is critical for reproductive success. Male juvenile exposure to other juveniles regardless of sex ratio caused significantly shorter adult longevity and faster decline in sperm ejaculation over successive matings. However, males from all treatments achieved similar number of matings in their lifetime. This study provides insight into adaptive resource allocation by males in response to juvenile sociosexual environment.     

Isozyme polymorphisms maintained by lethal loci in inbred strains of Aedes triseriatus

We analyzed the progeny of brother-sister matings of two inbred strains of the treehole mosquito, Aedes triseriatus, at several isozyme loci to determine which factors were maintaining heterozygosity at these loci after prolonged inbreeding. The triseriatus Kramer (TK) lines were polymorphic for the Odh and Hbd loci after 12 generations of full-sib mating, and the triseriatus Vero Beach (TV) lines were polymorphic for the Odh, Hk-4, Pgm, and Hbd loci after 14 generations of full-sib mating. Genetic analysis of 22 TK F13 matings and nine TV F15 matings showed that heterozygosity was enforced by lethal loci to which all the polymorphic isozyme loci were linked. The lethal loci formed a balanced lethal system. We determined the relative positions and map distances of lethals and enzyme loci.     

Correlations between fitness and heterozygosity at allozyme and microsatellite loci in the Atlantic salmon, Salmo salar L

The relationship between heterozygosity at genetic markers (six allozyme and eight microsatellite loci), and fluctuating asymmetry (FA), length and weight was investigated in two samples of Atlantic salmon (Salmo salar L.) with different timings of first active feeding (early (EA) and late (LA) salmon). This trait had previously been related to fitness. EA fish show smaller values of FA, are longer, heavier and are more heterozygous at allozyme loci than are conspecific LA fish. Also within both samples, heterozygosity at allozyme loci was inversely related to FA and was positively related to weight and length. However, no significant differences in microsatellite diversity (heterozygosity and mean d2 measurements) were observed between samples (EA vs LA). Furthermore, no association was observed between the variability at microsatellite loci and FA, weight or length within each sample. These results suggest that allozyme loci, in themselves, influence fitness components, rather than associations arising from associative overdominance.     

Context-Dependent Survival Differences among Electrophoretic Genotypes in Natural Populations of the Marine Bivalve Spisula Ovalis

We investigated the relationships between allozyme genotypes at nine polymorphic loci and survival in a natural population of the bivalve Spisula ovalis sampled on three occasions (1993, 1994, and 1995) in three different sites (2855 individuals analyzed). This species displays annual growth lines allowing identification of annual cohorts. Therefore we could avoid cohort mixing, a frequent bias in such studies, and evaluate the consistency of the observed effects across cohorts and sites. Significant viability differences were observed both among alleles and between heterozygotes and homozygotes at some loci. Multiple-locus heterozygosity was positively correlated with viability in the 1993-1994 period, but not in the 1994-1995 interval. The observed selective effects were significantly dependent on the cohort and the site considered. A bibliographic survey suggests that such variability is a common feature of studies analyzing heterozygosity-survival relationships. Two explanations are consistent with our results. First, allozyme genotypes may have direct effects on viability that interact with subtle environmental variation in a complex and unpredictable way. Second, allozyme genotypes may be transiently associated with other viability genes responsible for heterotic effects. In any case, the results militate against allozyme loci being themselves consistently overdominant for viability in natural populations.     

Variation in the effects of larval history on juvenile performance of a temperate reef fish

For organisms with complex life cycles, the transition between life stages can act as a significant demographic and selective bottleneck. Variation in developmental and growth rates among individuals present in one stage (e.g. larvae), due to initial differences in parental input and/or environmental conditions experienced, can propagate to future stages (e.g. juveniles), and such ‘carry‐over effects’ can shape fitness and phenotypic distributions within a population. However, variation in the strength of carry‐over effects between life stages and the intensity of selective mortality acting on intrinsic variation, and how these might be mediated by environmental variability in natural systems, is poorly known. Here, we evaluate variation in the strength to which larval growth histories can mediate juvenile performance (growth and survival), for a reef fish (Forsterygion lapillum) common to rocky reefs of New Zealand. We used otoliths to reconstruct demographic histories of recently settled fish that were sampled across cohorts, sites and microhabitats. We quantified sources of variation in the strength of carry‐over effects and selective mortality that operate on larval growth histories. We found overall that individuals that grew fast as larvae tended to experience proportional growth advantages as juveniles. However, the strength of growth advantages being maintained into the juvenile period varied among cohorts, sites and microhabitats. Specifically, a stronger growth advantage was found on some microhabitats (e.g. mixed stands of macroalgae) relative to others (e.g. monocultures of Carpophyllum maschalocarpum) for some cohorts and sites only. For other cohorts and sites, the degree of coupling between larval and juvenile growth rates was either indistinguishable between microhabitats or else not evident. Similarly, the intensity of growth‐based selective mortality varied among cohorts, sites and microhabitats: for the cohort and site where carry‐over effects differed between microhabitats, we also observed difference in the intensity to which fish with rapid larval growth rates were favoured. Overall, our results highlight how this spatial and temporal patchiness in extrinsic factors can interact with intrinsic variation of recruiting individuals to have a major influence on the resulting distribution of juveniles and their phenotypic traits.     

The impact of larval predators and competitors on the morphology and fitness of juvenile treefrogs

Studies of phenotypic plasticity typically focus on traits in single ontogenetic stages. However, plastic responses can be induced in multiple ontogenetic stages and traits induced early in ontogeny may have lasting effects. We examined how gray treefrog larvae altered their morphology in four different larval environments and whether different larval environments affected the survival, growth, development, and morphology of juvenile frogs at metamorphosis. We then reared these juveniles in terrestrial environments under high and low intraspecific competition to determine whether the initial differences in traits at metamorphosis affected subsequent survival and growth, whether the initial phenotypic differences converged over time, and whether competition in the terrestrial environment induced further phenotypic changes. Larval and juvenile environments both affected treefrog traits. Larval predators induced relatively deep tail fins and short bodies, but there was no impact on larval development. In contrast, larval competitors induced relatively short tails and long bodies, reduced larval growth, and slowed larval development. At metamorphosis, larval predators had no effect on juvenile growth or relative morphology while larval competitors produced juveniles that were smaller and possessed relatively shorter limbs and shorter bodies. After 1 month of terrestrial competition among the juvenile frogs, the initial differences in juvenile morphology did not converge. There were no differences in growth due to larval treatment but there were differences in survival. Individuals that experienced low competition as tadpoles experienced near perfect survival as juvenile frogs but individuals that experienced high competition as tadpoles suffered an 18% decrease in survival as juvenile frogs. There were also morphological responses to juvenile competition, but these changes appear to be due, at least in part, to allometric effects. Collectively, these results demonstrate that larval environments can have profound impacts on the traits and fitness of organisms later in ontogeny.     

Half-sib mating structures

All ways in which all matings in a population can be between half-sibs under a generalization of regular systems of inbreeding are characterized for both finite and infinite populations. A model of random half-sib mating is developed and analyzed, and the asymptotic configuration of populations subject to it is described. The classical model of half-sib mating which ensues from the standard definition of regular systems of inbreeding is only one of many ways a population can propagate by half-sib mating, and a wide range of genetic identity is possible dependent on which half-sib mating structure governs a population.     

Social common mole-rats enhance outbreeding via extra-pair mating

Females in many species engage in matings with males that are not their social mates. These matings are predicted to increase offspring heterozygosity and fitness, and thereby prevent the deleterious effects of inbreeding. We tested this hypothesis in a cooperative breeding mammal, the common mole-rat Cryptomys hottentotus hottentotus. Laboratory-based studies suggested a system of strict social monogamy, while recent molecular studies indicate extensive extra-pair paternity despite colonies being founded by an outbred pair. Our data show that extra-pair and within-colony breeding males differed significantly in relatedness to breeding females, suggesting that females may gain genetic benefits from breeding with non-resident males. Extra-colony male mating success was not based on heterozygosity criteria at microsatellite loci; however, litters sired by extra-colony males exhibited increased heterozygosity. While we do not have the data that refute a relationship between individual levels of inbreeding (Hs) and fitness, we propose that a combination of both male and female factors most likely explain the adaptive significance of extra-pair mating whereby common mole-rats maximize offspring fitness by detecting genetic compatibility with extra-pair mates at other key loci, but it is not known which sex controls these matings.     

Temperature influences selective mortality during the early life stages of a coral reef fish

For organisms with complex life cycles, processes occurring at the interface between life stages can disproportionately impact survival and population dynamics. Temperature is an important factor influencing growth in poikilotherms, and growth-related processes are frequently correlated with survival. We examined the influence of water temperature on growth-related early life history traits (ELHTs) and differential mortality during the transition from larval to early juvenile stage in sixteen monthly cohorts of bicolor damselfish Stegastes partitus, sampled on reefs of the upper Florida Keys, USA over 6 years. Otolith analysis of settlers and juveniles coupled with environmental data revealed that mean near-reef water temperature explained a significant proportion of variation in pelagic larval duration (PLD), early larval growth, size-at-settlement, and growth during early juvenile life. Among all cohorts, surviving juveniles were consistently larger at settlement, but grew more slowly during the first 6 d post-settlement. For the other ELHTs, selective mortality varied seasonally: during winter and spring months, survivors exhibited faster larval growth and shorter PLDs, whereas during warmer summer months, selection on PLD reversed and selection on larval growth became non-linear. Our results demonstrate that temperature not only shapes growth-related traits, but can also influence the direction and intensity of selective mortality.     

Characterization of microsatellite loci for the detection of temporal genetic shifts within a single cohort of the brown demoiselle,Neopomacentrus filamentosus

Neopomacentrus filamentosus, a common damselfish on the Indo–Australian archipelago, undergoes significant shifts in size and mitochondrial genetic structure upon larval settlement and metamorphosis to juvenile stages. We characterized five polymorphic microsatellite loci in order to study temporal genetic shifts within a single generation of N. filamentosus sampled first as larval settlers then again as demersal juvenile recruits. All loci were extremely polymorphic and exhibited high levels of heterozygosity. While all loci from the larval samples conformed to Hardy – Weinberg expectations, significant heterozygote deficiencies were seen in two loci in the juvenile samples, likely due to extreme size‐selective mortality imposed post‐settlement.     

Attempted use of an uncommon bay scallop color morph for tracking the contribution of restoration efforts to population recovery

In our ongoing bay scallop restoration efforts in eastern Long Island, NY, United States, we have planted millions of hatchery‐reared juveniles to serve as broodstock when they mature. These plantings have driven the resurgence of larval recruitment, benthic population sizes, and commercial fishery landings over a 12‐year period. In this article, we detail an attempt to track the contribution of our restoration efforts to population rebuilding using planted “skunks,” an uncommon color morph, by looking for increased skunk frequencies in subsequent wild cohorts. Although we saw higher skunk frequencies among large juveniles in natural populations in the first 3 years after extensive skunk plantings, supporting use of uncommon color morphs as a passive tracer, this conclusion was not supported when examined over a 12‐year period, as no strong correlations were seen between skunk frequencies at different life stages (adult broodstock, small and large F1 juveniles, F1 adults) for respective cohorts. Potential reasons for poor correlation between skunk frequencies and restoration efforts include lower than expected production of skunk offspring, dilution of out‐planted contribution to growing natural populations, interannual variability in skunk frequencies that may have obscured the expected skunk signal, and/or differentially higher mortality of skunks at postset and larger juvenile stages. In the latter case, skunks experienced higher overwinter mortality, most likely due to predation, in 9 of 11 years. This led us to suspend skunk plantings after 3 years. Nevertheless, commercial fishermen perceived skunks to be “our” scallops, helping raise the profile of and support for our restoration efforts.