Survival and growth of triploid eastern oysters, Crassostrea virginica, produced from wild diploids collected from low-salinity areas.

Triploid Eastern oysters have been reported to suffer greater mortalities than diploids when exposed to low-salinity (<5) conditions in the U.S. Gulf of Mexico and Atlantic estuaries. As such, the effect of broodstock parentage was investigated on the low-salinity tolerance of triploid progeny produced by mating diploid females (collected from three Louisiana estuaries differing in salinity regimes) with male tetraploids at two hatcheries. Diploid crosses were also produced using the wild broodstocks to verify expected differences in low-salinity tolerance among diploid progeny and between ploidy levels. All progeny were deployed at low and moderate-salinity (averages of 9.3 and 19.4) field sites to monitor monthly growth and mortality. Sex ratio, gametogenic stage, gonad-to-body ratio, condition index, and Perkinsus marinus infection were also measured periodically at both field sites Although high triploid mortality at the low-salinity site prevented complete analysis, results indicated that diploid parentage had little effect on triploid survival at low salinity. Broodstock parentage affected diploid mortality and growth, although results did not match with predictions made based on historical salinity at broodstock collection sites. Ploidy level had the largest effect on triploid survival and growth followed by the hatchery site where the oysters were produced.

Induced defenses as a management tool: Shaping individuals to their environment.

Many prey species can adjust morphology to reduce predation risk in response to predator cues. Enhancing prey defenses using predator cues may improve survival of cultivated species and enhance species restoration efforts, but assessment of such benefits at industrially relevant scales is needed. We examined how raising a model foundation species, oysters (Crassostrea virginica), under commercial hatchery conditions with cues from two common predator species can improve survival across a variety of predator regimes and environmental conditions. Oysters responded to predators by growing stronger shells than controls, but had subtle variations in shell characteristics depending on the predator species. Predator-induced changes significantly increased oyster survival up to 600% and survivorship was maximized when cue source was matched with local predator regime. Overall, our findings demonstrate the utility of using predator cues to enhance the survival of target species across landscapes and highlight the opportunity to employ nontoxic methods to control pest-based mortality.

Tolerance of northern Gulf of Mexico eastern oysters to chronic warming at extreme salinities.

The eastern oyster, Crassostrea virginica, provides critical ecosystem services and supports valuable fishery and aquaculture industries in northern Gulf of Mexico (nGoM) subtropical estuaries where it is grown subtidally. Its upper critical thermal limit is not well defined, especially when combined with extreme salinities. The cumulative mortalities of the progenies of wild C. virginica from four nGoM estuaries differing in mean annual salinity, acclimated to low (4.0), moderate (20.0), and high (36.0) salinities at 28.9 °C (84 °F) and exposed to increasing target temperatures of 33.3 °C (92 °F), 35.6 °C (96 °F) or 37.8 °C (100 °F), were measured over a three-week period. Oysters of all stocks were the most sensitive to increasing temperatures at low salinity, dying quicker (i.e., lower median lethal time, LT50) than at the moderate and high salinities and resulting in high cumulative mortalities at all target temperatures. Oysters of all stocks at moderate salinity died the slowest with high cumulative mortalities only at the two highest temperatures. The F1 oysters from the more southern and hypersaline Upper Laguna Madre estuary were generally more tolerant to prolonged higher temperatures (higher LT50) than stocks originating from lower salinity estuaries, most notably at the highest salinity. Using the measured temperatures oysters were exposed to, 3-day median lethal Celsius degrees (LD50) were estimated for each stock at each salinity. The lowest 3-day LD50 (35.1-36.0 °C) for all stocks was calculated at a salinity of 4.0, while the highest 3-day LD50 (40.1-44.0 °C) was calculated at a salinity of 20.0.

Eliciting expert judgment to inform management of diverse oyster resources for multiple ecosystem services.

This study presents the most comprehensive set of ecosystem service provision estimates for diverse oyster-based resources to date. We use expert elicitation methods to derive estimates of five ecosystem services provided by oysters: oyster harvest (as indicated by oyster density), improved water quality (net nitrogen assimilation), shoreline protection (net erosion), and other fish habitat (blue crab and red drum density). Distributions are estimated for three distinct resources: on-bottom production, off-bottom farms, and non-harvested restoration/conservation efforts, under twelve distinct scenarios according to varying environmental conditions (eutrophication, sedimentation, and salinity regimes). Our expert-derived estimates of ecosystem services provide useful comparisons across oyster resources of both expected ecosystem service delivery levels and the amount of variation in those levels. These estimates bridge an information gap regarding relative performance of diverse oyster resources along multiple dimensions and should serve as a useful guide for resource managers facing competing interests.

Effective reduction of Vibrio vulnificus in the Eastern oyster (Crassostrea virginica) using high salinity depuration.

Depuration under different salinities was used to reduce the human pathogen Vibrio vulnificus from Eastern oysters (Crassostrea virginica). Individual recirculating systems were used to test the efficacy of depuration at three salinities (15, 25, and 35 psu) in four independent trials during a 14 day period. Initial loads of V. vulnificus were higher than 10,000 MPN/g of oyster meat in all trials. Data showed that 25 and 35 psu treatments were more efficient in reducing V. vulnificus numbers than 15 psu with an overall reduction of >3 logs. A significant decrease in MPN/g was observed as early as day 6 and further reductions were observed at day 10, while longer depurations did not improve efficacy. Only the highest salinity (35 psu) was capable of reducing V. vulnificus numbers to the FDA recommended level of <30 MPN/g in two of the four trials. Oysters survived well in the depuration systems with minimal mortality (<1%) but their condition index (meat quality and yield) decreased during the 14 day period in all treatments. The data presented in this study suggests that high salinity depuration is a promising method to reduce V. vulnificus in oysters.

Breast cancer detection with upstream data fusion, machine learning, and automated registration: initial results.

Purpose: To develop an artificial intelligence algorithm for the detection of breast cancer by combining upstream data fusion (UDF), machine learning (ML), and automated registration, using digital breast tomosynthesis (DBT) and breast ultrasound (US). Approach: Our retrospective study included examinations from 875 women obtained between April 2013 and January 2019. Included patients had a DBT mammogram, breast US, and biopsy proven breast lesion. Images were annotated by a breast imaging radiologist. An AI algorithm was developed based on ML for image candidate detections and UDF for fused detections. After exclusions, images from 150 patients were evaluated. Ninety-five cases were used for training and validation of ML. Fifty-five cases were included in the UDF test set. UDF performance was evaluated with a free-response receiver operating characteristic (FROC) curve. Results: Forty percent of cases evaluated with UDF (22/55) yielded true ML detections in all three images (craniocaudal DBT, mediolateral oblique DBT, and US). Of these, 20/22 (90.9%) produced a UDF fused detection that contained and classified the lesion correctly. FROC analysis for these cases showed 90% sensitivity at 0.3 false positives per case. In contrast, ML yielded an average of 8.0 false alarms per case. Conclusions: An AI algorithm combining UDF, ML, and automated registration was developed and applied to test cases, showing that UDF can yield fused detections and decrease false alarms when applied to breast cancer detection. Improvement of ML detection is needed to realize the full benefit of UDF.

Assimilation of oil-derived elements by oysters due to the Deepwater Horizon Oil Spill.

During and after the Deepwater Horizon Oil Spill (DWHOS), oysters (Crassostrea virginica) were exposed to oil and susceptible to incidental consumption of surface and subsurface oil materials. We determined the contribution of oil materials from the DWHOS to diet of oysters by comparing carbon (C) and nitrogen (N) stable isotope ratios in oyster shell to ratios in suspended particulate matter (SPM) and in fresh and weathered oil. Average δ(13)C and δ(15)N values in oyster shell (-21 ± 1‰ and 9-11‰, respectively) were consistent with consumption of naturally available SPM as opposed to values in oil (-27 ± 0.2‰, 1.6 ± 0.4‰). Stable isotope ratios in oyster adductor muscle were similar to shell for δ(15)N but not δ(13)C, suggesting either a recent shift in diet composition or differential assimilation of C between tissue types. We found no evidence of assimilation of oil-derived C and N and, therefore, no evidence of an oyster-based conduit to higher trophic levels. Trace elements in shell were inconclusive to corroborate oil exposure. These findings are not an indication that oysters were not exposed to oil; rather they imply oysters either did not consume oil-derived materials or consumed too little to be detectable compared to natural diet.

Automated Registration for Dual-View X-Ray Mammography Using Convolutional Neural Networks.

OBJECTIVE: Automated registration algorithms for a pair of 2D X-ray mammographic images taken from two standard imaging angles, namely, the craniocaudal (CC) and the mediolateral oblique (MLO) views, are developed. METHODS: A fully convolutional neural network, a type of convolutional neural network (CNN), is employed to generate a pixel-level deformation field, which provides a mapping between masses in the two views. Novel distance-based regularization is employed, which contributes significantly to the performance. RESULTS: The developed techniques are tested using real 2D mammographic images, slices from real 3D mammographic images, and synthetic mammographic images. Architectural variations of the neural network are investigated and the performance is characterized from various aspects including image resolution, breast density, lesion size, lesion subtlety, and lesion Breast Imaging-Reporting and Data System (BI-RADS) category. Our network outperformed the state-of-the-art CNN-based and non-CNN-based registration techniques, and showed robust performance across various tissue/lesion characteristics. CONCLUSION: The proposed methods provide a useful automated tool for co-locating lesions between the CC and MLO views even in challenging cases. SIGNIFICANCE: Our methods can aid clinicians to establish lesion correspondence quickly and accurately in the dual-view X-ray mammography, improving diagnostic capability.

Effect of Gear Type on Vibrio spp. Levels in Farm-Raised Oysters (Crassostrea virginica) after Routine Handling and Resubmersion.

ABSTRACT: During routine handling, cultured oysters are removed from the water and exposed to elevated temperatures, causing growth of Vibrio vulnificus and Vibrio parahaemolyticus within them. Farmers can resubmerse oysters in the water, allowing elevated Vibrio spp. levels to return to ambient levels within the oysters. Previous resubmersion research is limited to one aquaculture gear type during studies performed from June to September. This study aims to expand knowledge about the recovery times needed for elevated Vibrio levels in handled oysters from two common gear types (the adjustable longline system and the OysterGro system) during early and midsummer periods. Oysters held in both gear types were subjected to being tumbled and refrigerated or desiccated and then resubmersed into water in May and July 2018 and 2019. Vibrio spp. levels were measured before and after the treatments, and 3, 7, and 14 days after resubmersion, and were compared with levels in submersed oysters. All samples were tested for V. vulnificus, total V. parahaemolyticus, and pathogenic V. parahaemolyticus (tdh+/trh+). Water temperatures in May were significantly lower (∼5°C; P ≤ 0.009) than in July, corresponding to lower V. vulnificus levels (-0.67 log MPN/g) and higher tdh+/trh+ levels (+0.56 to 0.63 log MPN/g) in control oysters. The average Vibrio spp. levels in control oysters from each trial did not differ between the gear types (P ≥ 0.08). Elevated V. vulnificus levels recovered to ambient levels after 7 days in May and 3 days in July, regardless of gear or handling. For V. parahaemolyticus, the desiccated oysters required 14 days to recover in May and 7 days in July, whereas the tumbled and refrigerated oysters required 14 days or more in both months. This study had limited replication in each month, but the data suggest that the resubmersion times differ between the gear types, treatment types, and months. Future studies with more replications are needed to determine whether these trends continue.

Sperm Repository for a Breeding Program of the Eastern Oyster Crassostrea virginica: Sample Collection, Processing, Cryopreservation, and Data Management Plan.

The Eastern oyster Crassostrea virginica (Family Ostreidae) is one of the most important fishery and aquaculture species in the U.S. and is a keystone species for coastal reefs. A breeding program was initiated in 2019 to support the fast-growing aquaculture industry culturing this species in the Gulf of Mexico. Oysters from 17 wild populations in embayment along the U.S. Gulf of Mexico coast from southwest Florida to the Matagorda Bay, Texas were used as broodstock for the program to maximize genetic diversity in the base population. A sperm repository of the broodstock was established to support the breeding project. The goal of this study was to demonstrate the sperm sample collection, processing, cryopreservation, and the data management plan involved in the establishment of a sperm germplasm repository of base populations. The supporting objectives were to: (1) develop a data management plan for the sperm repository; (2) streamline the procedure for sample collection, processing, and cryopreservation; (3) incorporate sperm quality analysis into the procedure, and (4) archive the cryopreserved samples as a repository for future use in the breeding program. This sperm repository included a total of 102 male oysters from the 17 collection sites (six oysters per site). A data management plan was developed with six categories, including sample collection, phenotype, fresh sperm, genotype, cryopreservation, and post-thaw sperm, as guide for data collection. Sperm collection was accomplished by strip spawn, and fresh sperm production, motility, and fertility were recorded for quality analysis. Cryopreserved sperm samples were sorted, labelled, archived, and stored in liquid nitrogen for future use. Post-thaw motility (1-30%) and plasm membrane integrity (15.34-70.36%) were recorded as post-thaw quality parameters. Overall, this study demonstrated a streamlined procedure of oyster sperm collection, processing, and cryopreservation for establishing a sperm repository that can serve as a template for construction of oyster germplasm repositories for breeding programs.

Effects of Farm Location on Vibrio parahaemolyticus and Vibrio vulnificus Levels in Oysters after Desiccation and Resubmersion in the Northern Gulf of Mexico.

ABSTRACT: Desiccation is a routine farming practice used in off-bottom oyster aquaculture to reduce biofouling organisms and improve shell quality. This practice can increase Vibrio parahaemolyticus and Vibrio vulnificus levels, leading to increased risk of illness for raw oyster consumers. Previous resubmersion studies were performed in geographic proximity to one another, so to better understand the broader applicability of resubmersion, the next step was to perform concurrent studies in multiple geographic locations within a region. This study evaluated the effect of variations in geographic location on the recovery time needed for elevated vibrio levels to return to ambient levels in desiccated oysters after resubmersion at Gulf Coast farms. Two trials were performed between May and August 2019 at sites spanning ∼100 km: three in Alabama and one in Florida. Oysters were deployed in OysterGro cages at each location, 2 weeks before each trial, and then either were desiccated for 24 h or remained submersed as controls. Triplicate samples were taken before and immediately following the desiccation period, as well as 7 and 14 days after resubmersion. Total and pathogenic V. parahaemolyticus and V. vulnificus levels were determined using most-probable-number (MPN) real-time PCR. Vibrio levels increased by 0.23 to 3.50 log MPN/g after desiccation. Recovery times varied among geographic locations by trial and Vibrio spp., with all vibrio counts recovering to levels not significantly higher than those in control oysters within 7 to 14 days of resubmersion (P ≥ 0.06). These results suggest a 14-day resubmersion period of cultured oysters allowed vibrio levels, elevated because of routine handling, to return to ambient levels at all farm sites studied.

Regulation of sperm motility in Eastern oyster (Crassostrea virginica) spawning naturally in seawater with low salinity.

Oyster aquaculture is expanding worldwide, where many farms rely on seed produced by artificial spawning. As sperm motility and velocity are key determinants for fertilization success, understanding the regulation of sperm motility and identifying optimal environmental conditions can increase fertility and seed production. In the present study, we investigated the physiological mechanisms regulating sperm motility in Eastern oyster, Crassostrea virginica. Sperm motility was activated in ambient seawater with salinity 4-32 PSU with highest motility and velocity observed at 12-24 PSU. In artificial seawater (ASW) with salinity of 20 PSU, sperm motility was activated at pH 6.5-10.5 with the highest motility and velocity recorded at pH 7.5-10.0. Sperm motility was inhibited or totally suppressed in Na+, K+, Ca2+, and Mg2+-free ASW at 20 PSU. Applications of K+ (500 µM glybenclamide and 10-50 mM 4-aminopyridine), Ca2+ (1-50 µM mibefradil and 10-200 µM verapamil), or Na+ (0.2-2.0 mM amiloride) channel blockers into ASW at 20 PSU inhibited or suppressed sperm motility and velocity. Chelating extracellular Ca2+ ions by 3.0 and 3.5 mM EGTA resulted in a significant reduction and full suppression of sperm motility by 4 to 6 min post-activation. These results suggest that extracellular K+, Ca2+, and Na+ ions are involved in regulation of ionic-dependent sperm motility in Eastern oyster. A comparison with other bivalve species typically spawning at higher salinities or in full-strength seawater shows that ionic regulation of sperm motility is physiologically conserved in bivalves. Elucidating sperm regulation in C. virginica has implications to develop artificial reproduction, sperm short-term storage, or cryopreservation protocols, and to better predict how changes in the ocean will impact oyster spawning dynamics.

Divergence in salinity tolerance of northern Gulf of Mexico eastern oysters under field and laboratory exposure.

The eastern oyster, Crassostrea virginica, is a foundation species within US Gulf of Mexico (GoM) estuaries that has experienced substantial population declines. As changes from management and climate are expected to continue to impact estuarine salinity, understanding how local oyster populations might respond and identifying populations with adaptations to more extreme changes in salinity could inform resource management, including restoration and aquaculture programs. Wild oysters were collected from four estuarine sites from Texas [Packery Channel (PC): 35.5, annual mean salinity, Aransas Bay (AB): 23.0] and Louisiana [Calcasieu Lake (CL): 16.2, Vermilion Bay (VB): 7.4] and spawned. The progeny were compared in field and laboratory studies under different salinity regimes. For the field study, F1 oysters were deployed at low (6.4) and intermediate (16.5) salinity sites in Alabama. Growth and mortality were measured monthly. Condition index and Perkinsus marinus infection intensity were measured quarterly. For the laboratory studies, mortality was recorded in F1 oysters that were exposed to salinities of 2.0, 4.0, 20.0/22.0, 38.0 and 44.0 with and without acclimation. The results of the field study and laboratory study with acclimation indicated that PC oysters are adapted to high-salinity conditions and do not tolerate very low salinities. The AB stock had the highest plasticity as it performed as well as the PC stock at high salinities and as well as Louisiana stocks at the lowest salinity. Louisiana stocks did not perform as well as the Texas stocks at high salinities. Results from the laboratory studies without salinity acclimation showed that all F1 stocks experiencing rapid mortality at low salinities when 3-month oysters collected at a salinity of 24 were used and at both low and high salinities when 7-month oysters collected at a salinity of 14.5 were used.

Effect of Ploidy on Vibrio parahaemolyticus and Vibrio vulnificus Levels in Cultured Oysters.

Vibrio parahaemolyticus and V. vulnificus are naturally occuring human pathogenic bacteria commonly found in estuarine environments where oysters are cultured. The use of triploid oysters has increased, due to their rapid growth rate and that they maintain a high quality throughout the year. Previous work suggested levels of Vibrio spp. may be lower in triploid oysters, as compared to diploids. Therfore, this study aimed to determine if there is a difference in the abundances of V. parahaemolyticus and V. vulnificus between half-sibling diploid and triploid oysters. In four trials, 100 individual oysters (either iced or temperature abused) were analyzed for V. parahaemolyticus and V. vulnificus using direct plating followed by colony hybridization. Mean levels of V. parahaemolyticus in iced and abused diploid oysters were 3.55 and 4.21 log CFU/g, respectively. Mean levels in iced and abused triploid oysters were 3.49 and 4.27 log CFU/g, respectively. Mean levels of V. vulnificus in iced and abused diploid oysters were 3.53 and 4.56 log CFU/g, respectively. Mean levels in iced and abused triploid oysters were 3.54 and 4.55 log CFU/g, respectively. The differences in Vibrio spp. abundances between diploid and triploid oysters was not significant (p>0.05). However, the differences across treatments were significant (p<0.05), with the exception of V. parahaemolyticus levels in trial 3 (p=0.83). Variation between individual oysters was also observed, with 12 of 808 measurements being outside of the 95 th percentile. This phenomenon of occasional statistical outliers ("hot" or "cold" oysters) has been previously described and supports the appropriateness of composite sampling to account for inherent animal variability. In summary, the data indicate that abundances of V. parahaemolyticus and V. vulnificus are not dependent on the ploidy of cultured oysters, but vary with the type of handling.

Effects of tumbling, refrigeration and subsequent resubmersion on the abundance of Vibrio vulnificus and Vibrio parahaemolyticus in cultured oysters (Crassostrea virginica).

Routine handling of oysters is a common industry practice for off-bottom oyster aquaculture, which aims to produce a high-quality oyster. These practices expose oysters to elevated temperatures and interrupt filter feeding, which can increase Vibrio vulnificus and V. parahaemolyticus levels within the oyster. The resubmersion of oysters after exposure to conditions where the time-temperature controls are exceeded is as an effective mitigation strategy to allow elevated levels of Vibrio spp. to "recover", or return to ambient levels, prior to harvest. Previous work examined the effect of desiccation on recovery times; the objective of this study was to evaluate the effect of additional handling treatments [tumbled and refrigerated (TR), tumbled and not refrigerated (TNR), not tumbled and refrigerated (NTR), and not tumbled and not refrigerated (NTNR)] on the time needed for V. vulnificus, total V. parahaemolyticus, and pathogenic V. parahaemolyticus (tdh+/trh+) to recover in oysters. A set of non-treated (control) oysters remained submerged throughout the study to determine the ambient Vibrio spp. (inclusive of genotypes) levels within oysters. Vibrio spp. levels were measured immediately before (pre) and after (post) the treatments, and 1, 2, 4, 7, 10, and 14 days after resubmersion using a three-tube MPN real-time PCR method. The non-refrigerated oysters (TNR, NTNR) had Vibrio spp. levels 1.54 to 2.10 log MPN/g higher than the pre-treatment levels, while the Vibrio spp. levels in refrigerated oysters were not significantly higher than pre-treatment levels. After resubmersion, Vibrio spp. levels increased by 0.84 to 1.78 log MPN/g in the refrigerated oysters (TR, NTR). Vibrio spp. levels in oysters returned to ambient after 1-7 days of resubmersion, depending on the handling treatment and the Vibrio spp. These results provide data on handling treatments not previously reported and further support the seven-day resubmersion requirement for farmers in Alabama using the adjustable longline system.

Towards CNN-Based Registration of Craniocaudal and Mediolateral Oblique 2-D X-ray Mammographic Images.

We investigate methodologies for the automated registration of pairs of 2-D X-ray mammographic images, taken from the two standard mammographic angles. We present two exploratory techniques, based on Convolutional Neural Networks, to examine their potential for co-registration of findings on the two standard mammographic views. To test algorithm performance, our analysis uses a synthetic, surrogate data set for performing controlled experiments, as well as real 2-D X-ray mammogram imagery. The preliminary results are promising, and provide insights into how the proposed techniques may support multi-view X-ray mammography image registration currently and as technology evolves in the future.

Effects of Desiccation Practices of Cultured Atlantic Oysters (Crassostrea virginica) on Vibrio spp. in Portersville Bay, Alabama, USA.

The expansion of off-bottom aquaculture to the Gulf of Mexico has raised public health concerns for human health officials. High temperatures in the Gulf of Mexico are associated with high levels of Vibrio parahaemolyticus and Vibrio vulnificus. Routine desiccation practices associated with off-bottom aquaculture expose oysters to ambient air, allowing Vibrio spp. to proliferate in the closed oyster. Currently, there is limited research on the length of time needed for Vibrio spp. levels in desiccated oysters to return to background levels, defined as the levels found in oysters that remain continually submersed and not exposed to ambient air. This study determined the time needed to return V. parahaemolyticus, V. vulnificus, and Vibrio cholerae levels to background levels in oysters exposed to the following desiccation practices: 3-h freshwater dip followed by 24-h ambient air exposure, 27-h ambient air exposure, and control. All oysters were submerged at least 2 weeks prior to the beginning of each trial, with the control samples remaining submerged for the duration of each trial. Vibrio spp. levels were enumerated from samples collected on days 0, 1, 2, 3, 7, 10, and 14 after resubmersion using a three-tube most-probable-number enrichment followed by BAX PCR. V. cholerae levels were frequently (92%) below the limit of detection at all times, so they were not statistically analyzed. V. parahaemolyticus and V. vulnificus levels in the 27-h ambient air exposure and the 3-h freshwater dip followed by 24-h ambient air exposure samples were significantly elevated compared with background samples. In most cases, the Vibrio spp. levels in oysters in both desiccation treatments remained elevated compared with background levels until 2 or 3 days post-resubmersion. However, there was one trial in which the Vibrio spp. levels did not return to background levels until day 7. The results of this study provide scientific support that oyster farmers should be required to implement a minimum 7-day resubmersion regimen. This length of time allowed the Vibrio spp. levels to become not significantly different across all treatments.

Transcriptomic evaluation of the American oyster, Crassostrea virginica, deployed during the Deepwater Horizon oil spill: Evidence of an active hydrocarbon response pathway.

Estuarine organisms were impacted by the Deepwater Horizon oil spill which released ∼5 million barrels of crude oil into the Gulf of Mexico in the spring and summer of 2010. Crassostrea virginica, the American oyster, is a keystone species in these coastal estuaries and is routinely used for environmental monitoring purposes. However, very little is known about their cellular and molecular responses to hydrocarbon exposure. In response to the spill, a monitoring program was initiated by deploying hatchery-reared oysters at three sites along the Alabama and Mississippi coast (Grand Bay, MS, Fort Morgan, AL, and Orange Beach, AL). Oysters were deployed for 2-month periods at five different time points from May 2010 to May 2011. Gill and digestive gland tissues were harvested for gene expression analysis and determination of aliphatic and polycyclic aromatic hydrocarbon (PAH) concentrations. To facilitate identification of stress response genes that may be involved in the hydrocarbon response, a nearly complete transcriptome was assembled using Roche 454 and Illumina high-throughput sequencing from RNA samples obtained from the gill and digestive gland tissues of deployed oysters. This effort resulted in the assembly and annotation of 27,227 transcripts comprised of a large assortment of stress response genes, including members of the aryl hydrocarbon receptor (AHR) pathway, Phase I and II biotransformation enzymes, antioxidant enzymes and xenobiotic transporters. From this assembly several potential biomarkers of hydrocarbon exposure were chosen for expression profiling, including the AHR, two cytochrome P450 1A genes (CYP1A-like 1 and CYP1A-like 2), Cu/Zn superoxide dismutase (CuZnSOD), glutathione S-transferase theta (GST theta) and multidrug resistance protein 3 (MRP3). Higher expression levels of GST theta and MRP3 were observed in gill tissues from all three sites during the summer to early fall 2010 deployments. Linear regression analysis indicated a statistically significant relationship between total PAH levels in digestive gland tissue samples with CYP1A-like 2, CuZnSOD, GST theta and MRP3 induction. These observations provide evidence of a potentially conserved AHR pathway in invertebrates and yield new insight into the development of novel biomarkers for use in environmental monitoring activities.

Preliminary study of transplanting as a process for reducing levels of Vibrio vulnificus and Vibrio parahaemolyticus in shellstock oysters.

Increasingly strict standards for harvest of oysters for the raw, half-shell market (designated as "white tag") should increase the proportion of oysters not meeting these standards (designated as "green tag"). Transplanting of green tag oysters into highsalinity waters (>20 practical salinity units) was explored as a means of returning Vibrio vulnificus and Vibrio parahaemolyticus levels to levels present on initial harvest. In summer 2011, oysters originally harvested in Louisiana were transplanted on two separate occasions (n = 2) to two sites in Mississippi Sound, AL: Sandy Bay and Dauphin Island. Oysters were tested for V. vulnificus and V. parahaemolyticus densities (by using the U.S. Food and Drug Administration enrichment method) after 2, 7, and 14 days deployed, with baseline samples taken (i) at the time of original harvest and iced, (ii) from oysters refrigerated within 1 h of harvest at <45°F ([7.2°C] white tag) and, (iii) from oysters not refrigerated during the harvest trip (green tag) but refrigerated after an 8-h trip. White and green tag oysters were sampled ∼24 h on arrival in Bon Secour, AL, put on ice, and shipped for analysis. Among baseline samples, there were no significant differences in V. vulnificus and V. parahaemolyticus densities, although the densities in the green tag oysters tended to be highest. After transplanting, V. vulnificus densities were significantly highest on day 2, with no significant differences among any of the other days within a site. On day 2, Sandy Bay had significantly greater densities of V. vulnificus than the Dauphin Island site, but no other days differed from time zero. For Vibrio parahaemolyticus, densities were greatest on day 2 and lowest at time zero, but this did not differ significantly from abundance on day 14. Average survival was 83.4% (± 3.13 SD), with no differences between sites. These preliminary results indicate that high-salinity transplanting could be an effective method of converting green tag oysters to oysters suitable for "reharvest" as white tag oysters.