• An assessment of RT-qPCR accuracy in monitoring infectious norovirus in oyster farms

      Hunt, K.; Butler, F.; Doré, B; Keaveney, S. (University College Dublin. School of Biosystems Engineering, 2014)
      Wastewater contamination causes Norovirus (NoV) to accumulate in commercial shellfish, which is monitored using RT-qPCR. RT-qPCR does not distinguish infectious copies from non-infectious, so it is not ideal for risk assessment. Additionally, sites being sampled are assumed to be spatially homogenous, but this has not been shown. To test homogeneity, and the standard sample size of ten, a single site was intensively sampled during the 2013- 2014 winter period. Analysis of results is ongoing. Also during the winter season, in three additional sites, the ratio of infectious to noninfectious virus copies detected with RT-qPCR was modelled using an FRNA bacteriophage surrogate. Analysis of results is ongoing.
    • Management of health risks associated with oysters harvested from a norovirus contaminated area, Ireland, February–March 2010

      Doré, B.; Keaveney, S.; Flannery, J.; Rajko-Nenow, P. (European Centre for Disease Prevention and Control, 2010)
      Oysters from a harvesting area responsible for outbreaks of gastroenteritis were relaid at a clean seawater site and subsequently depurated in tanks of purified seawater at elevated temperatures. This combined treatment reduced norovirus levels to those detected prior to the outbreak. On the basis of norovirus monitoring the sale of treated oysters was permitted although the harvest area remained closed for direct sale of oysters. No reports of illness have been associated with the consumption of treated oysters.
    • Norovirus genotypes present in oysters and in effluent from a wastewater treatment plant during the seasonal peak of infections in Ireland in 2010

      Rajow-Nenow, P.; Waters, A.; Keaveney, S.; Flannery, J.; Tuite, G.; Coughlan, S.; O’Flaherty, V.; Doré, W. (American Society for Microbiology, 2013)
      We determined norovirus (NoV) concentrations in effluent from a wastewater treatment plant and in oysters during the peak period of laboratory-confirmed cases of NoV infection in Ireland in 2010 (January to March). Weekly samples of influent, secondary treated effluent, and oysters were analyzed using real-time quantitative reverse transcription-PCR for NoV genogroup I (GI) and genogroup II (GII). The mean concentration of NoV GII (5.87 104 genome copies 100 ml 1) in influent wastewater was significantly higher than the mean concentration of NoV GI (1.40 104 genome copies 100 ml 1). The highest concentration of NoV GII (2.20 105 genome copies 100 ml 1) was detected in influent wastewater during week 6. Over the study period, a total of 931 laboratory-confirmed cases of NoV GII infection were recorded, with the peak (n 171) occurring in week 7. In comparison, 16 cases of NoV GI-associated illness were reported during the study period. In addition, the NoV capsid N/S domain was molecularly characterized for selected samples. Multiple genotypes of NoV GI (GI.1, GI.4, GI.5, GI.6, and GI.7) and GII (GII.3, GII.4, GII.6, GII.7, GII.12, GII.13, and GII.17), as well as 4 putative recombinant strains, were detected in the environmental samples. The NoV GII.4 variant 2010 was detected in wastewater and oyster samples and was the dominant strain detected in NoV outbreaks at that time. This study demonstrates the diversity of NoV genotypes present in wastewater during a period of high rates of NoV infection in the community and highlights the potential for the environmental spread of multiple NoV genotypes.
    • Preliminary results from a survey of oyster production areas in Ireland for norovirus

      Keaveney, S.; Flannery, J.; Guilfoyle, F.; Doré, J. (2007)
      A survey of 18 oyster production areas in Ireland for norovirus (NoV) contamination was initiated in August 2006. The findings presented are the preliminary results from the first seven months of the survey. Prior to the survey commencing, a simple desk bask sanitary survey of each area was undertaken. This provided an assessment enabling each site to be ranked into 3 categories (low, medium and high) on the basis of the risk of NoV contamination. Samples were collected on a monthly basis and tested for the presence of NoV using semi-quantitative real-time PCR allowing relative quantitation of NoV levels. A correlation was observed between occurrence and levels of NoV detected and the risk categories ascribed to each production area. To date NoV was detected in 60.7, 30.0 and 2.5 percent of samples from the high, medium and low risk categorised areas, respectively. A strong seasonal bias towards increased winter contamination was observed with NoV detected in 15.5 and 50 % of samples in August and February, respectively. The preliminary results from this survey indicate that it may be possible to predict the relative risk of NoV contamination in a shellfish harvesting area. This in conjunction with targeted NoV monitoring using real-time PCR could aid the further development of risk management procedures in shellfisheries.
    • Reproductive biology and parasite (Perkinsus marinus) prevalence in the eastern oyster, Crassostrea virginica, within a Georgia tidal river

      O'Beirn, F.X.; Walker, R.L.; Jansen, M.L. (Elisha Mitchell Scientific Society, 1997)
      Recruitment, in 1992, of the eastern oyster, Crassostrea virginica, at one site (Flume Dock) within the Sapelo Island National Estuarine Research Reserve (SINERR) was lower than other sampling sites. Recruitment allied with gametogenesis and parasite (Perkinsus marinus) prevalence within the oysters was examined in 1993, in an attempt to explain the differences observed the previous year. Sampling occurred at three sites (Marsh Landing, Jack Hammock, and Flume Dock) along the Duplin River within the SINERR. Oyster recruitment in 1993 was reduced in all sites within the SINERR. No oyster recruitment was recorded at the Flume Dock site during any monthly collections or on collectors deployed for the duration of the sampling study. Oysters developed gametogenically and spawned at the Marsh Landing site two weeks prior to their occurrence at the Jack Hammock site. Gametogenic development in oysters at the Flume Dock site was retarded a further two weeks where the males appeared to spawn six weeks after the females, which could account for reduced recruitment levels at this site. Approximately, twice as many females as males occurred in all sites. Monthly sampling of prevalence and intensity of Perkinsus marinus (Dermo) revealed high levels of both parameters. Smaller oysters did display lower prevalences of Dermo than larger forms.
    • Toward design criteria in constructed oyster reefs: oyster recruitment as a function of substrate type and tidal height

      O'Beirn, F.X.; Luchenbach, M.W.; Nestlerode, J.A.; Coates, G.M. (National Shellfisheries Association, 2000)
      Restoration of degraded oyster reef habitat generally begins with the addition of substrate that serves as a reef base and site for oyster spat attachment. Remarkably, little is known about how substrate type and reef morphology affect the development of oyster populations on restored reefs. Three-dimensional, intertidal reefs were constructed near Fisherman's Island, Virginia: two reefs in 1995 using surfclam (Spisula solidissima) shell and six reefs in 1996 using surfclam shell, oyster shell, and stabilized coal ash. We have monitored oyster recruitment and growth quarterly at three tidal heights (intertidal, mean low water, and subtidal) on each reef type since their construction. Oyster recruitment in 1995 exceeded that observed in the two subsequent years. High initial densities on the 1995 reefs decreased and stabilized at a mean of 418 oyster/m2. Oyster settlement occurred on all reef types and tidal heights in 1996; however, postsettlement mortality on the surfclam shell and coal ash reefs exceeded that on the oyster shell reefs, which remained relatively constant throughout the year (mean = 935 oysters/m2). Field observations suggest that predation accounts for most of the observed mortality and that the clam shell and coal ash reefs, which have little interstitial space, suffer greater predation. Oyster abundance was consistently greatest higher in the intertidal zone on all reefs in each year studied. The patterns observed here lead to the preliminary conclusion that the provision of spatial refugia (both intertidal and interstitial) from predation is an essential feature of successful oyster reef restoration in this region. In addition, high levels of recruitment can provide a numerical refuge, whereby the oysters themselves will provide structure and increase the probability of an oyster population establishing successfully on the reef.