Dermo Disease Research Articles

Understanding Crassostrea virginica tolerance of Perkinsus marinus through global gene expression analysis.

Disease tolerance, a host's ability to limit damage from a given parasite burden, is quantified by the relationship between pathogen load and host survival or reproduction. Dermo disease, caused by the protozoan parasite P. marinus, negatively impacts survival in both wild and cultured eastern oyster (C. virginica) populations. Resistance to P. marinus has been the focus of previous studies, but tolerance also has important consequences for disease management in cultured and wild populations. In this study we measured dermo tolerance and evaluated global expression patterns of two sensitive and two tolerant eastern oyster families experimentally challenged with distinct doses of P. marinus (0, 106, 107, and 108 parasite spores per gram wet weight, n = 3-5 individuals per family per dose). Weighted Gene Correlation Network Analysis (WGCNA) identified several modules correlated with increasing parasite dose/infection intensity, as well as phenotype. Modules positively correlated with dose included transcripts and enriched GO terms related to hemocyte activation and cell cycle activity. Additionally, these modules included G-protein coupled receptor, toll-like receptor, and tumor necrosis factor pathways, which are important for immune effector molecule and apoptosis activation. Increased metabolic activity was also positively correlated with treatment. The module negatively correlated with infection intensity was enriched with GO terms associated with normal cellular activity and growth, indicating a trade-off with increased immune response. The module positively correlated with the tolerant phenotype was enriched for transcripts associated with "programmed cell death" and contained a large number of tripartite motif-containing proteins. Differential expression analysis was also performed on the 108 dosed group using the most sensitive family as the comparison reference. Results were consistent with the network analysis, but signals for "programmed cell death" and serine protease inhibitors were stronger in one tolerant family than the other, suggesting that there are multiple avenues for disease tolerance. These results provide new insight for defining dermo response traits and have important implications for applying selective breeding for disease management.

Perkinsus marinus suppresses in vitro eastern oyster apoptosis via IAP-dependent and caspase-independent pathways involving TNFR, NF-kB, and oxidative pathway crosstalk

The protozoan parasite Perkinsus marinus causes Dermo disease in eastern oysters, Crassostrea virginica, and can suppress apoptosis of infected hemocytes using incompletely understood mechanisms. This study challenged hemocytes in vitro with P. marinus for 1 h in the presence or absence of caspase inhibitor Z-VAD-FMK or Inhibitor of Apoptosis protein (IAP) inhibitor GDC-0152. Hemocytes exposure to P. marinus significantly reduced granulocyte apoptosis, and pre-incubation with Z-VAD-FMK did not affect P. marinus-induced apoptosis suppression. Hemocyte pre-incubation with GDC-0152 prior to P. marinus challenge further reduced apoptosis of granulocytes with engulfed parasite, but not mitochondrial permeabilization. This suggests P. marinus-induced apoptosis suppression may be caspase-independent, affect an IAP-involved pathway, and occur downstream of mitochondrial permeabilization. P. marinus challenge stimulated hemocyte differential expression of oxidation-reduction, TNFR, and NF-kB pathways. WGCNA analysis of P. marinus expression in response to hemocyte exposure revealed correlated protease, kinase, and hydrolase expression that could contribute to P. marinus-induced apoptosis suppression.

A rapid phenotype change in the pathogen Perkinsus marinus was associated with a historically significant marine disease emergence in the eastern oyster

The protozoan parasite Perkinsus marinus, which causes dermo disease in Crassostrea virginica, is one of the most ecologically important and economically destructive marine pathogens. The rapid and persistent intensification of dermo in the USA in the 1980s has long been enigmatic. Attributed originally to the effects of multi-year drought, climatic factors fail to fully explain the geographic extent of dermo’s intensification or the persistence of its intensified activity. Here we show that emergence of a unique, hypervirulent P. marinus phenotype was associated with the increase in prevalence and intensity of this disease and associated mortality. Retrospective histopathology of 8355 archival oysters from 1960 to 2018 spanning Chesapeake Bay, South Carolina, and New Jersey revealed that a new parasite phenotype emerged between 1983 and 1990, concurrent with major historical dermo disease outbreaks. Phenotypic changes included a shortening of the parasite’s life cycle and a tropism shift from deeper connective tissues to digestive epithelia. The changes are likely adaptive with regard to the reduced oyster abundance and longevity faced by P. marinus after rapid establishment of exotic pathogen Haplosporidium nelsoni in 1959. Our findings, we hypothesize, illustrate a novel ecosystem response to a marine parasite invasion: an increase in virulence in a native parasite.

Survival and transcriptomic responses to different Perkinsus marinus exposure methods in an Eastern oyster family

The parasite Perkinsus marinus, which causes Dermo disease, affects Eastern oyster (Crassostrea virginica) survival and is responsible for substantial losses to aquaculture production. Controlled laboratory challenges enable definition and measurement of disease response phenotypes as well as characterization of gene expression patterns during the infection process. The method of parasite exposure and severity of infection likely affect response measures and exploring variation among them can enhance our understanding of this host-parasite interaction. Here we compared response traits and global expression patterns over time for a selectively bred Eastern oyster family exposed to P. marinus via either suspension feeding or direct injection to elucidate mechanisms determining survival. All exposed oysters (n = 56 per group) received a dose of 5 × 106 cultured P. marinus cells g−1 wet tissue weight but oysters in the injected group had higher initial parasite load and significantly reduced survival probability compared to the fed group. Using RNAseq, we identified differentially expressed transcripts between fed and control animals 6 h after exposure and between fed and injected groups at 36 h, 7 d, and 28 d post exposure. Immediately after infection, caspase-8 and transcripts associated with mediating cell damage from reactive oxygen species were upregulated in the fed group compared to unchallenged controls. The comparison between fed and injected groups early in the infection process revealed increased innate immune activity and overexpression of transcripts involved in proinflammatory cytokine pathways, including toll-like receptor 4 and TNF receptor-associated factor 6-like protein, in the injected group. Later in the infection process, the fed group expressed transcripts associated with growth, specifically microtubule-based processes, at a higher level than the injected group. This study demonstrates that dosing method influences initial parasite load, survival, and transcriptomic response to Dermo disease and highlights the importance of considering exposure method when interpreting interactions between P. marinus and the Eastern oyster.

A Dual Omics Approach to Evaluate Transcriptional and Metabolic Responses During Lipid Deprivation in an Oyster Parasite, Perkinsus marinus.

Perkinsus marinus, a protozoan and the causative agent of perkinsosis (dermo disease) is a prevalent parasite found within the eastern oyster (Crassostrea virginica). In this study, we explore metabolic processes of P. marinus cells under lipid-depleted medium conditions to elucidate the interchanging flux of lipid and carbohydrate metabolism. Although P. marinus can synthesize their own lipids from available nutrients, they display a slower growth in medium not supplemented with lipids as opposed to medium with lipids. Under these conditions, using transcriptomics, we surprisingly observed evidence of stimulated lipid degradation through increased transcription of two core β-oxidation pathway enzymes. Simultaneously, phospholipid biosynthetic pathways were downregulated. Metabolomic analysis supported the transcriptomic results. Most fatty acids were decreased in lipid-deplete medium as opposed to lipid-replete medium, and available glucose was fermented to lactate. A significant increase in the cholesterol derivative zymosterol further supported a downregulation of membrane synthesis under the experimental conditions. A robust tricarboxylic acid (TCA) cycle was apparent by enhanced citrate synthase transcription, and a simultaneous reduction in branched chain amino acids. It is concluded that although P. marinus has the capacity for synthesizing its own lipids, it can respond to lipid deprivation in medium by oxidizing readily available stores, and likely transitioning into a resting stage.

Environmental factors drive the release of Perkinsus marinus from infected oysters.

Since the discovery of Perkinsus marinus as the cause of dermo disease in Crassostrea virginica, salinity and temperature have been identified as the main environmental drivers of parasite prevalence. However, little is known about how these variables affect the movement of the parasite from host to water column. In order to elucidate how environmental factors can influence the abundance of this parasite in the water column, we conducted a series of experiments testing the effects of time of day, temperature and salinity on the release of P. marinus cells from infected oysters. We found that P. marinus cells were released on a diurnal cycle, with most cells released during the hottest and brightest period of the day (12:00-18:00). Temperature also had a strong and immediate effect on the number of cells released, but salinity did not, only influencing the intensity of infection over the course of several months. Taken together, our results demonstrate that (1) the number of parasites in the water column fluctuates according to a diurnal cycle, (2) temperature and salinity act on different timescales to influence parasite abundance, and (3) live infected oysters may substantially contribute to the abundance of transmissive parasites in the water column under particular environmental conditions.

Factors Affecting Dermo Disease (Perkinsus marinus) in Eastern Oysters (Crassostrea virginica) in Galveston Bay, Texas

Factors Affecting Dermo Disease (Perkinsus marinus) in Eastern Oysters (Crassostrea virginica) in Galveston Bay, Texas

Variation in global transcriptomic response to Perkinsus marinus infection among eastern oyster families highlights potential mechanisms of disease resistance

Dermo disease, caused by the protozoan parasite Perkinsus marinus, negatively impacts wild and cultured Eastern oyster populations, yet our knowledge of the mechanistic bases for parasite pathogenicity and the Eastern oyster's response to it is limited. To better understand host responses to the parasite and identify molecular mechanisms underlying disease-resistance phenotypes, we experimentally challenged two families exhibiting divergent Dermo-resistance phenotypes with the parasite, generated global expression profiles using RNAseq and identified differentially expressed transcripts between control and challenged oysters from each family at multiple time points post-parasite injection. The susceptible and resistant families exhibited strikingly different transcriptomic responses to the parasite over a 28-day time period. The resistant family exhibited a strong, focused, early response to P. marinus infection, where many significantly upregulated transcripts were associated with the biological processes “regulation of proteolysis” and “oxidation-reduction process.” P. marinus virulence factors are mainly comprised of proteases that facilitate parasite invasion and weaken host humoral defenses, thus host upregulation of transcripts associated with negative regulation of proteolysis is consistent with a Dermo-resistant phenotype. In contrast, the susceptible family mounted a very weak, disorganized, initial response to the parasite. Few transcripts were differentially expressed between control and injected oysters, and no functional enrichment was detected among them. At the final 28 d time point 2450 differentially expressed transcripts were identified and were associated with either “G-protein coupled receptor activity” (upregulated) or “microtubule-based process” (downregulated). A handful of protease inhibitors were differentially expressed between control and injected susceptible oysters, but this function was not enriched in the susceptible data set. The differential expression patterns observed in this study provide valuable insight into the functional basis of Dermo resistance and suggest that the timing of expression is just as important as the transcripts being expressed.

Defining Dermo resistance phenotypes in an eastern oyster breeding population

Perkinsus marinus, the causative agent of Dermo disease, is responsible for mass mortalities and negatively impacts aquaculture production of the eastern oyster, Crassostrea virginica. Selective breeding is a viable option for Dermo disease management; however, fluctuations in natural selection pressure and environmental noise hinder accumulation of genetic gains acquired through field performance trials. The purpose of this study was to adapt and apply laboratory disease challenge methods to eastern oysters, better characterize resistance-specific traits and assess the potential for genetic variation in Dermo resistance among distinct families within a breeding population. Two challenge experiments were conducted, one in 2014 and the other in 2015. Significant treatment (control vs. challenged) and family effects on survival (measured as per cent survival and days to death) were detected in the 2014 challenge, while overall high survival precluded the detection of a significant family effect in the 2015 challenge. An alternate measure of resistance, parasite elimination rate, was also measured in the 2015 challenge, and this varied significantly among families. Thus, both survival and the change in parasite concentration in oyster tissues over time represent Dermo resistance phenotypes that can be measured accurately with the adapted laboratory disease challenge protocol described here. The obvious next step is to incorporate the challenge protocol in eastern oyster breeding programmes to assess whether well-defined, accurately measured, Dermo-resistant phenotypes result in enhanced genetic improvement for this commercially important trait.

Modeling the transmission of Perkinsus marinus in the Eastern oyster Crassostrea virginica

Dermo disease caused by the protistan Perkinsus marinus in Eastern oysters Crassotrea virginica is an important source of mortality impacting oyster population dynamics resulting in substantial losses in fisheries and aquaculture. The rapid transmission and spread of the disease minimized the importance of transmission models and past models (proliferation-based models) assumed simple density-dependent transmission or rapid infection post-settlement. This approach is a good approximation only for low population densities. A transmission model was developed for P. marinus in Eastern oysters that accounts for the seasonal change in disease dynamics and density-dependent foraging (of suspended particles) interference among hosts. The model, verified and evaluated against field observations, incorporates parasite release to the water column from live and dead individuals, parasite consumption by living oysters, the diffusion of parasites in the water, body burden-based dose-dependent transmission, recruitment, and disease-caused mortality. The model returns a basic reproduction number R0 for Dermo much greater than unity (R0=90) in accordance with the current persistence and pandemic nature of this disease in oysters. No population density is obtained that is low enough to suppress R0 below 1 (i.e. disease extinction). R0 is also estimated for high oyster densities (>300 individuals m−2) and particularly for relatively large oysters (∼90mm), today rare but once common before generalized overfishing occurred on healthy oyster reefs. In this scenario, R0 drops below 1, indicating that high oyster density can limit disease invasion through foraging interference and depletion of parasites in the water column. High intensity recruitment events allow the oyster population to attain such densities and limit the development of epizootics. These results provide insight into the transition from past populations, where Dermo is inferred to have been limited in its impact, to the current persistent and pandemic nature of this disease. Further coupling of this model into metapopulation and hydrodynamic models could be a promising tool to support management decision-making for bivalve populations impacted by Dermo disease.

What Is Going on with Perkinsus marinus in the Gulf of Mexico?

The National Oceanic and Atmospheric Administration (NOAA) Status and Trends Mussel Watch Program sampled the largest oysters in nearly every major US coastal lagoon and estuary in the Gulf of Mexico once during the winter from 1986 to 2010. This contribution examines trends in the principal oyster disease in the Gulf of Mexico, Dermo, caused by Perkinsus marinus, and some related population dynamic characteristics for its host, Crassostrea virginica. During the 1986–2000 period, P. marinus prevalence and infection intensity and oyster population dynamics followed the El Nino-Southern Oscillation (ENSO) cycle, responding to variations in salinity caused by variations in rainfall and freshwater inflow. The ENSO signal in the oyster population effectively ceased circa 2002. Beginning around this time, wintertime P. marinus prevalence and weighted prevalence began a decadal decline, as did the length of the largest oysters and the fraction of these largest animals that were female. The trends in Dermo disease, oyster length, and oyster sex ratio are all consistent with the following hypothesis: increasing temperature during the 2000s resulted in an increase in P. marinus infection intensity sufficient to increase the mortality rate in late summer and fall in the larger animals. This simultaneously reduced Dermo prevalence and infection intensity in the winter at the time of sampling and also resulted in the decline in the length of the largest animals targeted by Mussel Watch. Coincident with the decline in length is the expected decline in the fraction female, such that the percent female in the largest animals dropped to ≤50 % throughout much of the Gulf of Mexico. The decline in length leading to fewer large animals reproducing and the loss of females are predicted to have reduced oyster population reproductive capacity substantially during the 2000s. The early 60 % of the Mussel Watch time series took place during a period of negative Atlantic Multidecadal Oscillation (AMO) indices. The AMO moved into positive territory circa 2000. A positive AMO index is consistent with observed warmer water temperatures, and increased water temperature is consistent with an increase in Dermo-induced mortality.

Epizootiology of Perkinsus marinus, parasite of the pleasure oyster Crassostrea corteziensis, in the Pacific coast of Mexico

The protozoan parasite Perkinsus marinus is the etiological agent of “dermo disease”. This pathogen is considered by the World Organization for Animal Health (OIE) as reportable due to the high mortalities that it produces in the eastern oyster Crassostrea virginica in the US. In 2006, this parasite was detected in the pleasure oyster Crassostrea corteziensis in Nayarit on the Pacific coast of Mexico, indicating a new host and an extension of its known distribution. Epizootiological data of P. marinus in the pleasure oyster are unknown. With the objective of determining the prevalence and intensity in relation with temperature and salinity throughout time, as well as for studying interactions of host size and sex with the parasite, a monthly sampling was carried out in two aquaculture sites of Nayarit from 2007 to 2014. A total of 7700 oysters were analyzed. In both localities, prevalence was low in winter (<6%) when temperature and salinity fluctuated around 24°C and 33, respectively; and the highest prevalence values occurred during summer (37%) when temperature and salinity were around 30°C and 20, respectively. Infection intensity increased in summer, but severe cases remained on average <10%. Larger oysters showed the highest prevalence and intensity, and higher prevalence were generally observed in females. No unusual mortalities directly related with P. marinus were observed.

Effects of air-exposure gradients on spatial infection patterns of Perkinsus marinus in the eastern oyster Crassostrea virginica.

Spatial distributions of species can be shaped by factors such as parasites, mortality, and reproduction, all of which may be influenced by differences in physical factors along environmental gradients. In nearshore tidal waters, an elevational gradient in aerial exposure during low tide can shape the spatial distributions of benthic marine organisms. The eastern oyster Crassostrea virginica is an ecologically and economically important species that can dominate both subtidal and intertidal habitats along the east coast of the USA. Our goal was to determine whether prevalence and intensity of Perkinsus marinus (the causative agent of Dermo disease) infections vary along intertidal to subtidal gradients during summer. We used (1) field experiments conducted at 4 sites in the Chesapeake Bay and a Virginia coastal bay, (2) a controlled air-exposure experiment, and (3) field surveys from 7 sites ranging from Maine to North Carolina to test for effects of tidal exposure on infection. Results from our field surveys suggested that high intertidal oysters tend to have higher infection prevalence than subtidal oysters, but there was no effect on infection intensity. Field experiments rarely yielded significant effects of tidal exposure on infection prevalence and intensity. Overall, our study shows that exposure to air may not be a strong driver of infection patterns in this host-parasite system.

Anthropogenic impacts on an oyster metapopulation: Pathogen introduction, climate change and responses to natural selection

Abstract Humans rely on marine ecosystems for a variety of services but often impact these ecosystems directly or indirectly limiting their capacity to provide such services. One growing impact is the emergence of marine disease. We present results from a unique case study examining how oysters, a dominant organism in many coastal bays and estuaries that is often harvested for food, have responded to pathogens influenced by human activities, namely the introduction of novel pathogens. Climate change has enabled a northward spread and establishment of Dermo disease in oysters along the eastern seaboard of North America and human activities inadvertently introduced MSX disease along this same coast. Oysters in Delaware Bay have responded differently to each pathogen, and uniquely to MSX disease by developing a highly resistant baywide population not documented in any other bay. Offspring were produced using parents collected from low or high disease (MSX and Dermo) regions of Delaware Bay and exposed in a common garden experiment along with a naïve population from Maine. Results indicated widespread resistance to MSX disease, but not to Dermo disease, across Delaware Bay. One striking result was the demonstration of resilience in the population through its capacity to spread, presumably through larval transport, resistance to MSX disease into portions of the population that have experienced little to no MSX disease pressure themselves. Related studies indicated that larval transport mechanisms allowed widespread dispersal such that the entire metapopulation could acquire a high level of resistance over time if disease resistance is sufficiently heritable. The findings have implications for restoration, management and recovery of diseased populations. Namely, that if left to their own devices, natural selection may find a solution that enables populations to recover from introduced pathogens.

Effects of co-varying diel-cycling hypoxia and pH on disease susceptibility in the eastern oyster Crassostrea virginica

MEPS Marine Ecology Progress Series Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections MEPS 538:169-183 (2015) - DOI: https://doi.org/10.3354/meps11479 Effects of co-varying diel-cycling hypoxia and pH on disease susceptibility in the eastern oyster Crassostrea virginica Andrew G. Keppel1,2,*, Denise L. Breitburg1, Gary H. Wikfors3, Rebecca B. Burrell1, Virginia M. Clark1,2 1Smithsonian Environmental Research Center, Edgewater, MD 21037, USA 2Marine Estuarine and Environmental Sciences, University of Maryland, College Park, MD 20742, USA 3NOAA Fisheries Service, Northeast Fisheries Science Center, Milford, CT 06460, USA *Corresponding author: keppela@si.edu ABSTRACT: Diel-cycling hypoxia co-occurs with diel-cycling pH in shallow waters that are typically considered as refuge from deep-water hypoxia and are, therefore, targeted for restoration. These areas also tend to be heavily impaired by eutrophication from nutrient over-enrichment which increases the occurrence and severity of hypoxia and pH cycles. We used laboratory experiments to investigate the effects of diel-cycling dissolved oxygen (DO) and co-varying diel-cycling pH on infections of Perkinsus spp. and hemocyte activity in the eastern oyster Crassostrea virginica. Perkinsus marinus is the protistan parasite that causes Dermo disease in oysters. Severe diel-cycling DO increased the acquisition and progression of Perkinsus infections during exposure, and had a legacy effect the next year. Diel-cycling pH did not significantly affect infection dynamics either on its own or in combination with diel-cycling DO. Diel-cycling DO and pH both individually and in conjunction stimulated hemocyte activity, although this stimulated activity may not be effective at preventing Perkinsus infection. The magnitude of cycling conditions is an important consideration when choosing restoration sites, as severe cycling may hinder the re-establishment of oysters by creating areas that serve as reservoirs for parasites that can infect nearby populations. KEY WORDS: Hypoxia · pH · Diel cycle · Multiple stressors · Disease · Oyster Full text in pdf format Erratum PreviousNextCite this article as: Keppel AG, Breitburg DL, Wikfors GH, Burrell RB, Clark VM (2015) Effects of co-varying diel-cycling hypoxia and pH on disease susceptibility in the eastern oyster Crassostrea virginica. Mar Ecol Prog Ser 538:169-183. https://doi.org/10.3354/meps11479 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in MEPS Vol. 538. Online publication date: October 28, 2015 Print ISSN: 0171-8630; Online ISSN: 1616-1599 Copyright © 2015 Inter-Research.

The influence of site and season on the gut and pallial fluid microbial communities of the eastern oyster, Crassostrea virginica (Bivalvia, Ostreidae): community-level physiological profiling and genetic structure

The microbial communities and overall health of the eastern oyster, Crassostrea virginica, have long been topics of interest due to the fundamental economic and ecological roles this species maintains. A broad scale characterization of the oyster microbiome over spatial and seasonal scales, however, has never been carried out. The primary goal of this study was to examine the factors mediating microbial communities of the gut and pallial fluid of C. virginica at three sites within the Long Island Sound estuary, with a focus on both genetic structure (T-RFLP) and physiological profiling (EcoPlates) of the microbiome. Results indicated that the genetic structure of microbial communities of oysters was minimally separated across sites, but was influenced by season. Although the microbial community structure was similar, the number of carbon sources utilized by these communities (richness) varied across site, season, and anatomical location within the host. Parameters including oyster condition index, Dermo disease, and ambient water temperature were measured to assess their influence on the oyster microbiome. Only water temperature was found to have a significant relationship with microbial community structure and richness. Results suggest that a core microbiome may exist within the eastern oyster, specifically for those populations that are not genetically distinct.

Physiological and pathological changes in the eastern oyster Crassostrea virginica infested with the trematode Bucephalus sp. and exposed to the toxic dinoflagellate Alexandrium fundyense

Effects of experimental exposure to Alexandrium fundyense, a Paralytic Shellfish Toxin (PST) producer known to affect bivalve physiological condition, upon eastern oysters, Crassostrea virginica with a variable natural infestation of the digenetic trematode Bucephalus sp. were determined.After a three-week exposure to cultured A. fundyense or to a control algal treatment with a non-toxic dinoflagellate, adult oysters were assessed for a suite of variables: histopathological condition, hematological variables (total and differential hemocyte counts, morphology), hemocyte functions (Reactive Oxygen Species (ROS) production and mitochondrial membrane potential), and expression in gills of genes involved in immune responses and cellular protection (MnSOD, CAT, GPX, MT-IV, galectin CvGal) or suspected to be (Dominin, Segon).By comparing individual oysters infested heavily with Bucephalus sp. and uninfested individuals, we found altered gonad and digestive gland tissue and an inflammatory response (increased hemocyte concentration in circulating hemolymph and hemocyte infiltrations in tissues) associated with trematode infestation. Exposure to A. fundyense led to a higher weighted prevalence of infection by the protozoan parasite Perkinsus marinus, responsible for Dermo disease. Additionally, exposure to A. fundyense in trematode-infested oysters was associated with the highest prevalence of P. marinus infection. These observations suggest that the development of P. marinus infection was advanced by A. fundyense exposure, and that, in trematode-infested oysters, P. marinus risk of infection was higher when exposed to A. fundyense. These effects were associated with suppression of the inflammatory response to trematode infestation by A. fundyense exposure. Additionally, the combination of trematode infestation and A. fundyense exposure caused degeneration of adductor muscle fibers, suggesting alteration of valve movements and catch state, which could increase susceptibility to predation. Altogether, these results suggest that exposure of trematode-infested oysters to A. fundyense can lead to overall physiological weakness that decrease oyster defense mechanisms.

Microsatellite Genotypes Reveal Some Long-Distance Gene Flow inPerkinsus marinus, a Major Pathogen of the Eastern Oyster,Crassostrea virginica(Gmelin)

ABSTRACT As the agent of Dermo disease, Perkinsus marinus causes significant mortality and reduced fecundity its eastern oyster host, Crassostrea virginica. Passive dispersal of P. marinus between hosts subjects parasite movements to control by water currents in estuarine systems, potentially limiting connectivity among parasite populations in different estuaries. Given recent evidence for sexual reproduction in P. marinus, estimates of gene flow among locations may provide insights into this parasite's epidemiology. In this study, 1,082 wild oysters were collected from 2002 to 2008 at 15 geographical locations encompassing 4,800 km of the Gulf of Mexico and Atlantic coasts of the United States. Of these, 742 oysters (68.9% prevalence) were determined to be infected by P. marinus. Among infected oysters, 374 were determined to be infected by a single parasite genotype based on amplification of seven microsatellite loci, and therefore were amenable to analysis. Allele frequencies differed significantly amo...

Hybridization between previously isolated ancestors may explain the persistence of exactly two ancient lineages in the genome of the oyster parasite Perkinsus marinus

Theory predicts that neutral genetic variation accumulates within populations to a level determined by gains through mutation and losses by genetic drift. This balance results in a characteristic distribution of allelic variation with the maximum allelic difference determined by effective population size. Here, we report a striking departure from these expectations in the form of allelic dimorphism, observed at the majority of seven loci examined in Perkinsus marinus, an important oyster parasite that causes Dermo disease. DNA sequences were collected from five loci flanking microsatellite repeats and two loci coding for superoxide dismutase enzymes that may mediate the parasite’s interaction with its host. Based on 474 sequences, sampled across 5000km of the eastern United States coastline, no more than two alleles were observed at each locus (discounting singletons). Depending on the locus, the common allele ranged in overall frequency from 72% to 92%. At each locus the two alleles differed substantially (3.8% sequence difference, on average), and the among-locus variance in divergences was not sufficient to reject a simultaneous origin for all dimorphisms using approximate Bayesian methods. Dimorphic alleles were estimated to have diverged from a common ancestral allele at least 0.9 million years ago. Across these seven loci, only five other alleles were ever observed, always as singletons and differing from the dimorphic alleles by no more than two nucleotides. Free recombination could potentially have shuffled these dimorphisms into as many as 243 multilocus combinations, but the existence of only ten combinations among all samples strongly supports low recombination frequencies and is consistent with the observed absence of intragenic recombination. We consider several demographic and evolutionary hypotheses to explain these patterns. Few can be conclusively rejected with the present data, but we advance a recent hybridization of ancient divergent lineages scenario as the most parsimonious.

Hemocytes and Plasma of the Eastern Oyster (Crassostrea virginica) Display a Diverse Repertoire of Sulfated and Blood Group A-modified N-Glycans*

The eastern oyster (Crassostrea virginica) has become a useful model system for glycan-dependent host-parasite interactions due to the hijacking of the oyster galectin CvGal1 for host entry by the protozoan parasite Perkinsus marinus, the causative agent of Dermo disease. In this study, we examined the N-glycans of both the hemocytes, which via CvGal1 are the target of the parasite, and the plasma of the oyster. In combination with HPLC fractionation, exoglycosidase digestion, and fragmentation of the glycans, mass spectrometry revealed that the major N-glycans of plasma are simple hybrid structures, sometimes methylated and core α1,6-fucosylated, with terminal β1,3-linked galactose; a remarkable high degree of sulfation of such glycans was observed. Hemocytes express a larger range of glycans, including core-difucosylated paucimannosidic forms, whereas bi- and triantennary glycans were found in both sources, including structures carrying sulfated and methylated variants of the histo-blood group A epitope. The primary features of the oyster whole hemocyte N-glycome were also found in dominin, the major plasma glycoprotein, which had also been identified as a CvGal1 glycoprotein ligand associated with hemocytes. The occurrence of terminal blood group moieties on oyster dominin and on hemocyte surfaces can account in part for their affinity for the endogenous CvGal1.