Assessment of Infection Prevalence and Intensity of Disease-Causing Parasitic Protozoans Perkinsus marinus and Haplosporidium nelsoni in Georgia Oysters.

Eastern oysters, Crassostrea virginica, are ecologically and economically important coastal species which provide a commercially valuable food product while also improving water quality through filtration, protecting shorelines, and providing habitat. The protozoan parasites Perkinsus marinus and Haplosporidium nesloni commonly infect oysters along the United States Atlantic and Gulf coasts and have been linked to poor oyster health and mass mortality events. In this study, wild oysters were collected from multiple reefs within four tidal creeks along the coast of Georgia to investigate P. marinus and H. nelsoni prevalence and intensity, their potential impact on oyster health, and identify possible drivers of the parasites. A second study occurred on four sites on Sapelo Island, Georgia, with continuous water quality monitoring data to further elucidate potential drivers. Oyster density and condition index, a proxy for health, were measured, and parasites were quantified using a TaqMan probe based quantitative real-time PCR within gill tissue. Real-time PCR showed that 86% of oysters tested were infected by one or both parasites in the coast-wide survey, and 93% of oysters from Sapelo Island were also infected by one or both parasites. Prevalence and infection intensity for both P. marinus and H. nelsoni varied across sites. Overall impacts on oysters were complex-intensity was not linked to oyster metrics in the coastwide study, but oyster condition was negatively correlated with P. marinus prevalence in the Sapelo Island study. Several relationships between both parasites and water quality parameters were identified, providing valuable information about potential drivers that should be investigated further.

Strong male-biased operational sex ratio in a breeding population of loggerhead turtles (Caretta caretta) inferred by paternal genotype reconstruction analysis.

Characterization of a species mating systems is fundamental for understanding the natural history and evolution of that species. Polyandry can result in the multiple paternity of progeny arrays. The only previous study of the loggerhead turtle (Caretta caretta) in the USA showed that within the large peninsular Florida subpopulation, multiple paternity occurs in approximately 30% of clutches. Our study tested clutches from the smaller northern subpopulation for the presence of multiple paternal contributions. We examined mothers and up to 20 offspring from 19.5% of clutches laid across three nesting seasons (2008-2010) on the small nesting beach on Wassaw Island, Georgia, USA. We found that 75% of clutches sampled had multiple fathers with an average of 2.65 fathers per nest (1-7 fathers found). The average number of fathers per clutch varied among years and increased with female size. There was no relationship between number of fathers and hatching success. Finally, we found 195 individual paternal genotypes and determined that each male contributed to no more than a single clutch over the 3-year sampling period. Together these results suggest that the operational sex ratio is male-biased at this site.

Interpopulation hybridization results in widespread viability selection across the genome in Tigriopus californicus.

BACKGROUND: Genetic interactions within hybrids influence their overall fitness. Understanding the details of these interactions can improve our understanding of speciation. One experimental approach is to investigate deviations from Mendelian expectations (segregation distortion) in the inheritance of mapped genetic markers. In this study, we used the copepod Tigriopus californicus, a species which exhibits high genetic divergence between populations and a general pattern of reduced fitness in F2 interpopulation hybrids. Previous studies have implicated both nuclear-cytoplasmic and nuclear-nuclear interactions in causing this fitness reduction. We identified and mapped population-diagnostic single nucleotide polymorphisms (SNPs) and used these to examine segregation distortion across the genome within F2 hybrids. RESULTS: We generated a linkage map which included 45 newly elucidated SNPs and 8 population-diagnostic microsatellites used in previous studies. The map, the first available for the Copepoda, was estimated to cover 75% of the genome and included markers on all 12 T. californicus chromosomes. We observed little segregation distortion in newly hatched F2 hybrid larvae (fewer than 10% of markers at p < 0.05), but strikingly higher distortion in F2 hybrid adult males (45% of markers at p < 0.05). Hence, segregation distortion was primarily caused by selection against particular genetic combinations which acted between hatching and maturity. Distorted markers were not distributed randomly across the genome but clustered on particular chromosomes. In contrast to other studies in this species we found little evidence for cytonuclear coadaptation. Instead, different linkage groups exhibited markedly different patterns of distortion, which appear to have been influenced by nuclear-nuclear epistatic interactions and may also reflect genetic load carried within the parental lines. CONCLUSION: Adult male F2 hybrids between two populations of T. californius exhibit dramatic segregation distortion across the genome. Distorted loci are clustered within specific linkage groups, and the direction of distortion differs between chromosomes. This segregation distortion is due to selection acting between hatching and adulthood.

Tracing hybrid incompatibilities to single amino acid substitutions.

Deleterious interactions among genes cause reductions in fitness of interpopulation hybrids (hybrid breakdown). Identifying genes involved in hybrid breakdown has proven difficult, and few studies have addressed the molecular basis of this widespread phenomenon. Because proper function of the mitochondrial electron transport system (ETS) requires a coadapted set of nuclear and mitochondrial gene products, ETS genes present an attractive system for studying the evolution of coadapted gene complexes within isolated populations and the loss of fitness in interpopulation hybrids. Here we show the effects of single amino acid substitutions in cytochrome c (CYC) on its functional interaction with another ETS protein, cytochrome c oxidase (COX) in the intertidal copepod Tigriopus californicus. The individual and pairwise consequences of three naturally occurring amino acid substitutions in CYC are examined by site-directed mutagenesis and found to differentially effect the rates of CYC oxidation by COX variants from different source populations. In one case, we show that interpopulation hybrid breakdown in COX activity can be attributed to a single naturally occurring amino acid substitution in CYC.

Evolution, biogeography, and the utility of mitochondrial 16s and COI genes in phylogenetic analysis of the crab genus Austinixa (Decapoda: Pinnotheridae).

This study used molecular data (mitochondrial 16s and COI) for the first time to explore evolutionary relationships among species of the pinnotherid crab genus Austinixa. Low levels of phylogenetic signal were detected for COI. High levels of phylogenetic signal were detected for 16s, indicating it is a more useful marker for inferring species level phylogenies in Austinixa. Phylogeographic patterns among species of Austinixa are consistent with allopatric speciation due to numerous climatic and oceanographic fluctuations during the last 5-6 my. In addition, all but two species have been derived since the closure of the Isthmus of Panama, a pattern consistent with hypotheses that the marine biota of the Caribbean and southeastern North America underwent a pulse of biotic turnover within the last 2-3 my. Austinixa aidae and Austinixa hardyi had identical 16s sequences, and differed by only 2 bp in COI, raising questions about the validity of A. hardyi as a distinct species.

Molecular and quantitative trait variation within and among populations of the intertidal copepod Tigriopus californicus.

While molecular and quantitative trait variation may be theoretically correlated, empirical studies using both approaches frequently reveal discordant patterns, and these discrepancies can contribute to our understanding of evolutionary processes. Here, we assessed genetic variation in six populations of the copepod Tigriopus californicus. Molecular variation was estimated using five polymorphic microsatellite loci, and quantitative variation was measured using 22-life history and morphometric characters. Within populations, no correlation was found between the levels of molecular variation (heterozygosity) and quantitative variation (heritability). Between populations, quantitative subdivision (Q(ST)) was correlated with molecular subdivision when measured as F(ST) but not when measured as R(ST). Unlike most taxa studied to date, the overall level of molecular subdivision exceeded the level of quantitative subdivision (F(ST) = 0.80, R(ST) = 0.89, Q(ST) = 0.30). Factors that could contribute to this pattern include stabilizing or fluctuating selection on quantitative traits or accelerated rates of molecular evolution.