Prevalence and characterization of Salmonella serovars isolated from oysters served raw in restaurants

To determine if Salmonella-contaminated oysters are reaching consumer tables, a survey of raw oysters served in eight Tucson restaurants was performed from October 2007 to September 2008. Salmonella spp. were isolated during 7 of the 8 months surveyed and were present in 1.2% of 2,281 oysters tested. This observed prevalence is lower than that seen in a previous study in which U.S. market oysters were purchased from producers at bays where oysters are harvested. To test whether the process of refrigerating oysters in restaurants for several days reduces Salmonella levels, oysters were artificially infected with Salmonella and kept at 4°C for up to 13 days. Direct plate counts of oyster homogenate showed that Salmonella levels within oysters did not decrease during refrigeration. Six different serovars of Salmonella enterica were found in the restaurant oysters, indicating multiple incidences of Salmonella contamination of U.S. oyster stocks. Of the 28 contaminated oysters, 12 (43%) contained a strain of S. enterica serovar Newport that matched by pulsed-field gel electrophoresis a serovar Newport strain seen predominantly in the study of bay oysters performed in 2002. The repeated occurrence of this strain in oyster surveys is concerning, since the strain was resistant to seven antimicrobials tested and thus presents a possible health risk to consumers of raw oysters.     

Depuration and Relaying: A Review on Potential Removal of Norovirus from Oysters

Pollution of coastal waters can result in contamination of bivalve shellfish with human enteric viruses, including norovirus (NoV), and oysters are commonly implicated in outbreaks. Depuration is a postharvest treatment involving placement of shellfish in tanks of clean seawater to reduce contaminant levels; this review focuses on the efficacy of depuration in reducing NoV in oysters. There have been many NoV outbreaks from depurated oysters containing around 10³ genome copies/g oyster tissue, far exceeding the median infectious dose (ID50). Half of the published NoV reduction experiments showed no decrease in NoV during depuration, and in the remaining studies it took between 9 and 45.5 d for a 1‐log reduction—significantly longer than commercial depuration time frames. Surrogate viruses are more rapidly depurated than NoV; the mean number of days to reduce NoV by 1 log is 19, and 7.5 d for surrogates. Thus, surrogates do not appear to be suitable for assessing virological safety of depurated oysters; data on reduction of NoV infectivity during depuration would assist evaluations on surrogate viruses and the impact of methods used. The longer persistence of NoV highlights its special relationship with oysters, which involves the binding of NoV to histo‐blood group‐like ligands in various tissues. Given the persistence of NoV and on‐going outbreaks, depuration as currently performed appears ineffective in guaranteeing virologically safe oysters. Conversely, relaying oysters for 4 wk is more successful, with low NoV concentrations and no illnesses associated with products. The ineffectiveness of depuration emphasizes the need for coastal water quality to be improved to ensure oysters are safe to eat.     

福建省养殖环节牡蛎中诺如病毒污染状况分析

目的 了解福建省养殖环节牡蛎中诺如病毒(Norovirus)污染状况及基因分型,进一步定量分析诺如病毒污染水平,为食品中诺如病毒污染监测及安全风险评估提供数据支持。方法 2017年8月—2018年9月,于福建省某养殖基地采集牡蛎样品共244份,采用实时荧光定量聚合酶链式反应(PCR)法对样品中诺如病毒进行检测,确定其污染情况及基因分型。结果 211份有效样品中诺如病毒阳性率为30.33%(64/211),GI和GII组诺如病毒均有检出。GI组诺如病毒阳性率为16.59%(35/211),GII组阳性率为24.17%(51/211),GI和GII组同时为阳性的比例为10.43%(22/211)。诺如病毒含量在1.05×10²～1.68×10⁴基因拷贝/g(消化腺)之间。结论 福建省养殖环节牡蛎样品中存在诺如病毒污染,冬季阳性率较高,提示有关部门需在冬季采取更多针对性的风险管理干预措施,以降低食源性诺如病毒感染风险。     

Ice immersion as a postharvest treatment of oysters for the reduction of Vibrio vulnificus

Vibrio vulnificus produces serious illnesses that are commonly associated with shellfish consumption, particularly raw oysters. Ingestion can result in fatal septicemia in susceptible individuals with hepatitis, cirrhosis, immune dysfunction, diabetes, or hemochromatosis (metabolic iron overload). Therefore, postharvest treatments to reduce vibrio levels in oysters have been recommended. In this study, rapid chilling by immersion of unwashed whole oysters in ice for 3 h was assessed as a postharvest treatment for reduction of V. vulnificus. Treated oysters were subsequently refrigerated at 45 degrees F (7.2 degrees C), whereas control oysters were not iced but were maintained at 45 degrees F throughout the study. Homogenized meats were monitored for total heterotrophic aerobic bacteria, V. vulnificus, and fecal coliform content before and after treatment over a 2-week period. V. vulnificus was enumerated by DNA probe hybridization of colonies from standard plate counts on nonselective medium, and recovery was compared for several media. Loss of plating efficiency was observed on standard selective and differential media compared with nonselective agars. Numbers of V. vulnificus generally declined in treated samples compared with controls; however, increases in total heterotrophic bacteria and fecal coliforms were also observed in treated samples at some time points. This study does not support the use of ice immersion as a postharvest method because of the relatively small declines in V. vulnificus numbers and the possibility of concomitant increases in fecal coliform and total bacterial contamination.     

季节性自回归差分移动平均模型在牡蛎中诺如病毒检出率预测上的应用

目的 基于季节性自回归差分移动平均(ARIMA)模型分析并预测上海市售牡蛎中诺如病毒(NoV)的检出率,为水产品中NoV的污染规律提供参考。方法 2016年6月—2019年11月,从上海芦潮港海鲜市场定期采购牡蛎样品共531只,通过巢式聚合酶链式反应(Nest-PCR),对其进行了NoV检测,按季度分析检出率。采用季节性ARIMA模型对牡蛎中NoV的检出率数据拟合建立模型,经过数据平稳化、模型选择和拟合、模型诊断得到最优模型,并运用最优模型对未来四个季度牡蛎中NoV的检出率进行预测。结果 拟合出的季节性ARIMA(0,1,1)(0,1,0)₄为最优模型,赤池信息量准则的修正值(AICc)最小(58.70),残差经Ljung-Box检验为白噪声序列。模型拟合牡蛎中NoV的阳性率趋势与实际检出率趋势基本吻合,平均绝对误差(MAE)为4.85,平均绝对百分比误差(MAPE)为30.25。用最优模型预测的未来四个季度牡蛎中NoV阳性检出率分别为31.89%、12.80%、9.47%、6.14%。结论 季节性ARIMA模型(0,1,1)(0,1,0)₄能较好地拟合牡蛎中NoV的阳性检出率趋势,对NoV污染的牡蛎等水产品的风险评估及NoV流行的防控具有一定的意义。     

Inactivation of a calicivirus and enterovirus in shellfish by high pressure

This study demonstrates the potential of high pressure (HP) processing to reduce viral contamination in shellfish. Bovine enterovirus, which is structurally similar to hepatitis A virus, was more pressure-resistant than feline calicivirus, a surrogate for norovirus. Both viruses were more pressure-resistant when treated in “naturally” contaminated mussels and oysters, compared to seawater and culture medium, suggesting that the medium can have a significant protective effect against HP treatment. Treatment at pressures of 250 MPa showed only a limited inactivation of either virus in shellfish, suggesting that relatively mild HP treatments (approximately 260 MPa) currently used for the commercial processing of oysters, principally to assist the shucking process, may be insufficient to ensure the safety of shellfish for human consumption, particularly in relation to human pathogenic viruses.Industrial relevanceHigh pressure (HP) processing is increasingly being used in the commercial processing of oysters, principally to assist the opening or shucking of oysters. Little is known about the effect of HP treatment on pathogens in shellfish, particularly human enteric viruses, which are the predominant cause of shellfish-borne disease. This article demonstrates the inactivation of surrogate animal viruses, as potential models for noroviruses and hepatitis A virus, by HP processing and compares the efficacy of inactivation when viruses are treated in culture medium, seawater and shellfish. The potential of HP processing to reduce viral contamination in shellfish is discussed in light of these findings.     

2020~2021年广州市售牡蛎中GII型诺如病毒污染情况调查

滤食性牡蛎是食源性诺如病毒传播的重要食品媒介。为了解广州市售牡蛎中的诺如病毒污染水平与遗传多样性特点,合理评估消费风险,本研究于2020年6月至2021年5月期间,每月从当地水产市场随机采集牡蛎样本,采用实验室前期建立的蛋白酶K处理偶联聚乙二醇沉淀小体系法,包括荧光定量RT-PCR和巢式RT-PCR技术检测贝类中病毒的污染量以及基因型分布特点。结果共检测牡蛎110只,GII型诺如病毒阳性检出率为52.7%（58/110）,病毒污染含量范围为1.56×103~1.09×106 copies/g（消化腺）。其中,春夏季节（3~8月）牡蛎中诺如病毒的阳性率为35.7%（20/56）,低于在秋冬季节（9~2月）的阳性率70.4%（38/54）;但不同季节中检出的病毒含量无显著差异,分别为春季（2.69±1.46）×105 copies/g（消化腺）,夏季（1.97±2.16）×105 copies/g（消化腺）,秋季（6.91±6.16）×104 copies/g（消化腺）,冬季为（4.83±2.99）×104 copies/g（消化腺）。部分阳性样本测序分析后显示,除1份为GII.17基因型外,其余均为GII.4基因型（n=13）,与当地的临床流行基因型呈现一致性。本研究显示广州市售牡蛎中仍存在较高的诺如病毒污染水平,需要进一步加强病毒防控工作,尤其提醒消费者在食用牡蛎时需加工充分。     

Cryptosporidium and Giardia in commercial and non-commercial oysters (Crassostrea gigas) and water from the Oosterschelde, The Netherlands

The intestinal parasites Cryptosporidium and Giardia cause gastro-enteritis in humans and can be transmitted via contaminated water. Oysters are filter feeders that have been demonstrated to accumulate pathogens such as Salmonella, Vibrio, norovirus and Cryptosporidium from contaminated water and cause foodborne infections. Oysters are economically important shellfish that are generally consumed raw. Commercial and non-commercial oysters (Crassostrea gigas) and oyster culture water from the Oosterschelde, The Netherlands, were examined for the presence of Cryptosporidium oocysts and Giardia cysts. Nine of 133 (6.7%) oysters from two non-commercial harvesting sites contained Cryptosporidium, Giardia or both. Six of 46 (13.0%) commercial oysters harboured Cryptosporidium or Giardia in their intestines. Data on the viability of (oo)cysts recovered from Oosterschelde oysters were not obtained, however viable (oo)cysts were detected in surface waters that enter the Oosterschelde oyster harvesting areas. The detection of Cryptosporidium and Giardia in oysters destined for human consumption has implications for public health only when human pathogenic (oo)cysts that have preserved infectivity during their stay in a marine environment are present. Our data suggest that consumption of raw oysters from the Oosterschelde may occasionally lead to cases of gastro-intestinal illness.     

北京市市售带壳牡蛎致病性弧菌污染状况调查

目的了解北京市市售带壳牡蛎致病性弧菌污染状况。方法 2014年2~11月每月在某水产品批发市场的摊位抽样200只带壳牡蛎,共80份样品(其中腮和肠样品分别为40份)。用常规培养方法检测牡蛎腮和肠(含便)中致病性弧菌,对副溶血性弧菌进行血清学分型,荧光定量PCR检测副溶血性弧菌毒力基因tdh、trh和tlh。结果 80份牡蛎样品中,致病性弧菌阳性样品检出率为62.50%(50/80),副溶血性弧菌阳性菌株检出率为33.75%(27/80),溶藻弧菌阳性菌株检出率为31.25%(25/80);各牡蛎腮和肠样品中,致病性弧菌阳性检出率为67.50%(27/40)和57.50%(23/40);27株副溶血性弧菌共9种血清型;毒力基因检测结果表示,tlh均为阳性,tdh和trh均为阴性。结论北京市市售带壳牡蛎中致病性弧菌污染严重,以副溶血性弧菌和溶藻弧菌检出为主。     

Vibrio vulnificus and Vibrio parahaemolyticus in U.S. retail shell oysters: a national survey from June 1998 to July 1999.

From June 1998 to July 1999, 370 lots of oysters in the shell were sampled at 275 different establishments (71%, restaurants or oyster bars; 27%, retail seafood markets: and 2%, wholesale seafood markets) in coastal and inland markets throughout the United States. The oysters were harvested from the Gulf (49%). Pacific (14%), Mid-Atlantic (18%), and North Atlantic (11%) Coasts of the United States and from Canada (8%). Densities of Vibrio vulnificus and Vibrio parahaemolyticus were determined using a modification of the most probable number (MPN) techniques described in the Food and Drug Administration's Bacteriological Analytical Manual. DNA probes and enzyme immunoassay were used to identify suspect isolates and to determine the presence of the thermostable direct hemolysin gene associated with pathogenicity of V. parahaemolyticus. Densities of both V. vulnifcus and V. parahaemolyticus in market oysters from all harvest regions followed a seasonal distribution, with highest densities in the summer. Highest densities of both organisms were observed in oysters harvested from the Gulf Coast, where densities often exceeded 10,000 MPN/g. The majority (78%) of lots harvested in the North Atlantic, Pacific, and Canadian Coasts had V. vulnificus densities below the detectable level of 0.2 MPN/g; none exceeded 100 MPN/g. V. parahaemolyticus densities were greater than those of V. vulnificus in lots from these same areas, with some lots exceeding 1,000 MPN/g for V. parahaemolyticus. Some lots from the Mid-Atlantic states exceeded 10,000 MPN/g for both V. vulnificus and V. parahaemolyicus. Overall, there was a significant correlation between V. vulificus and V. parahaemolyticus densities (r = 0.72, n = 202, P < 0.0001), but neither density correlated with salinity. Storage time significantly affected the V. vulnificus (10% decrease per day) and V. parahaemolyticus (7% decrease per day) densities in market oysters. The thermostable direct hemolysin gene associated with V parahaemolyticus virulence was detected in 9 of 3,429 (0.3%) V. parahaemolyticus cultures and in 8 of 198 (4.0%) lots of oysters. These data can be used to estimate the exposure of raw oyster consumers to V. vulnificus and V. parahaemolyticus.     

Effects of electrolyzed oxidizing water treatment on reducing Vibrio parahaemolyticus and Vibrio vulnificus in raw oysters

Contamination of Vibrio parahaemolyticus and Vibrio vulnificus in oysters is a food safety concern. This study investigated effects of electrolyzed oxidizing (EO) water treatment on reducing V. parahaemolyticus and V. vulnificus in laboratory-contaminated oysters. EO water exhibited strong antibacterial activity against V. parahaemolyticus and V. vulnificus in pure cultures. Populations of V. parahaemolyticus (8.74 x 10(7) CFU/ml) and V. vulnificus (8.69 x 10(7) CFU/ml) decreased quickly in EO water containing 0.5% NaCl to nondetectable levels (> 6.6 log reductions) within 15 s. Freshly harvested Pacific oysters were inoculated with a five-strain cocktail of V. parahaemolyticus or V. vulnificus at levels of 10(4) and 10(6) most probable number (MPN)/g and treated with EO water (chlorine, 30 ppm; pH 2.82; oxidation-reduction potential, 1131 mV) containing 1% NaCl at room temperature. Reductions of V. parahaemolyticus and V. vulnificus in oysters were determined at 0 (before treatment), 2, 4, 6, and 8 h of treatment. Holding oysters inoculated with V. parahaemolyticus or V. vulnificus in the EO water containing 1% NaCl for 4 to 6 h resulted in significant (P < 0.05) reductions of V. parahaemolyticus and V. vulnificus by 1.13 and 1.05 log MPN/g, respectively. Extended exposure (> 12 h) of oysters in EO water containing high levels of chlorine (> 30 ppm) was found to be detrimental to oysters. EO water could be used as a postharvest treatment to reduce Vibrio contamination in oysters. However, treatment should be limited to 4 to 6 h to avoid death of oysters. Further studies are needed to determine effects of EO water treatment on sensory characteristics of oysters.     

Geographical,temporal, and species variation of the polyether toxins,azaspiracids, in shellfish

Azaspiracid Poisoning (AZP) is a new toxic syndrome that has caused human intoxications throughout Europe following the consumption of mussels (Mytilus edulis), harvested in Ireland. Shellfish intoxication is a consequence of toxin-bearing microalgae in the shellfish food chain, and these studies demonstrated a wide geographic distribution of toxic mussels along the entire western coastal region of Ireland. The first identification of azaspiracids in other bivalve mollusks including oysters (Crassostrea gigas), scallops (Pecten maximus), clams (Tapes phillipinarium), and cockles (Cardium edule) is reported. Importantly, oysters were the only shellfish that accumulated azaspiracids at levels that were comparable with mussels. The highest levels of total azaspiracids (microg/g) recorded to-date were mussels (4.2), oysters (2.45), scallops (0.40), cockles (0.20), and clams (0.61). An examination of the temporal variation of azaspiracid contamination of mussels in a major shellfish production area revealed that, although maximum toxin levels were recorded during the late summer period, significant intoxications were observed at periods when marine dinoflagellate populations were low. Although human intoxications have so far only been associated with mussel consumption, the discovery of significant azaspiracid accumulation in other bivalve mollusks could pose a threat to human health.     

Detection of norovirus RNA in bivalve molluscs by using bacteria-culture-employed method (A3T method)

Norovirus (NV) RNA has rarely been detected in foods despite the use of highly sensitive methods such as RT-PCR and real-time RT-PCR. In the modified method (A3T method) reported previously, a bacterial culture process was introduced into the standard protocol for NV detection to remove some inhibitor(s) present in food ingredients. To confirm the efficiency of the A3T method and to examine NV contamination in bivalve molluscs, we tried to detect NV RNA in bivalve molluscs on the market and in oyster samples associated with foodborne outbreaks by using the standard method and the A3T method. NV RNAs were detected in 20 samples (18.0%) of 111 bivalve molluscs, including oysters, on the market by use of the A3T method, while only one sample (0.9%) was positive according to the standard method. NV RNA was also detected in 10 of 35 oyster samples related to foodborne outbreaks by the A3T method. Those results show that the A3T method is suitable for the detection of NV in bivalve molluscs in general laboratories.     

Fatty acids,volatile compounds and colour changes in high-pressure-treated oysters (Crassostrea gigas)

Proximal composition, colour and levels of fatty acids and volatile compounds in oysters following high-pressure (HP) treatment at 260, 500 or 800 MPa for 3, 5 or 5 min, respectively, were investigated and compared to untreated oysters. HP-treated oysters had significantly (P < 0.05) higher pH than untreated oysters. HP treatment also modified the gross composition of oyster tissue, the moisture content of HP-treated oyster being higher than that of untreated oyster. HP treatment at 260 MPa had less negative effects on oyster tissue colour (Hunter L-, a- and b-values) than treatment at higher pressures. HP treatment of oysters resulted in no significant changes in the fatty acid profile compared to untreated oysters. HP treatment of oysters, however, changed the level of volatile components when compared to the headspace of fresh oysters; HP-treated samples had higher concentrations of dimethyl sulfide, 1-penten-3-one, phenol and 1,2,4-trimethylbenzene relative to untreated samples. On the other hand, HP-treated oysters had lower concentrations of 1-penten-3-ol, 2,3-pentanedione, (E,E,Z)-1,3,5-octatriene and 1,3-octadiene than untreated samples.Industrial relevanceHigh-pressure (HP) treatment is being increasingly employed for commercial processing of oysters. While the profiles of fatty acids and volatiles of fresh oysters have been extensively studied, no studies reporting the effects of HP on these have been reported. Compared to fresh oysters, HP treatment did not change significantly the fatty acid profile; however, differences in the headspace volatile profile of HP-treated oysters were noticed.     

Comparative Toxicity and Paralytic Shellfish Poisoning Toxin Profiles in the Mussel Mytilus galloprovincialis and the Oyster Crassostrea gigas Collected from a Mediterranean Lagoon in Tunisia: A Food Safety Concern

Edible shellfish Mytilus galloprovincialis and Crassostrea gigas have been investigated for the paralytic shellfish poisons using mouse bioassay and high performance liquid chromatography with fluorescence detection. Paralytic shellfish poisons toxins were detected in mussels and oysters from September 2007 to May 2008. The level of paralytic shellfish poisons toxins in mussels reached the maximum in November with 832.9 μg saxitoxin-eq/100 g tissue. In oysters, toxins were detected with a maximum of 11.2 μg saxitoxin-eq/100 g tissue. The toxin high performance liquid chromatography profiles in mussels and oysters revealed the dominance of gonyautoxin 5 and N-sulfocarbamoyl-gonyautoxin-2 and -3 (C1-2), whereas GTX1-4, saxitoxin, and neosaxitoxin were found at low amounts. Overall, levels of paralytic shellfish poisons toxins were 20–70 times greater in mussels than in oysters. This is the first report on the qualitative and quantitative paralytic shellfish poisons content of M. galloprovincialis and C. gigas from a shellfish farming lagoon in Tunisia.     

Behavior and incidence of Vibro parahaemolyticus in Sydney rock oysters (Crassostrea commercialis)

Vibrio parahaemolyticus grew poorly or not at all during storage of unopened Sydney rock oysters (Crassostrea commercialis) at 15 and 30°C for 2 and 7 days. Although V. parahaemolyticus counts often increased at 30°C, counts above 10⁴/g were not observed. Escherichia coli counts did not usually change substantially under these conditions. V. parahaemolyticus grew more readily during storage of unopened oysters under more severe conditions, with counts approaching or exceeding 10⁶/g after continuous or intermittent storage at 37°C. Opened oysters provided a much more favourable environment than unopened oysters for growth of V. parahaemolyticus. Growth occurred at 15, 30 and 37°C, with counts > 10⁶/g after overnight storage at 30 or 37°C. A survey of 30 market samples of oysters was conducted. Sixteen samples of unopened oysters were collected at the wholesale level and 14 samples of refrigerated opened oysters were purchased from retailers. V. parahaemolyticus was present in all samples of unopened oysters (range 0.4/g?2.3 × 10⁴/g) and in 13 samples of opened oysters (range 4.3/g? > 1.1 × 10³/g).     

Managing the risk of Vibrio parahaemolyticus infections associated with oyster consumption: A review

Vibrio parahaemolyticus is a Gram‐negative bacterium that is naturally present in the marine environment. Oysters, which are water filter feeders, may accumulate this pathogen in their soft tissues, thus increasing the risk of V. parahaemolyticus infection among people who consume oysters. In this review, factors affecting V. parahaemolyticus accumulation in oysters, the route of the pathogen from primary production to consumption, and the potential effects of climate change were discussed. In addition, intervention strategies for reducing accumulation of V. parahaemolyticus in oysters were presented. A literature review revealed the following information relevant to the present study: (a) managing the safety of oysters (for human consumption) from primary production to consumption remains a challenge, (b) there are multiple factors that influence the concentration of V. parahaemolyticus in oysters from primary production to consumption, (c) climate change could possibly affect the safety of oysters, both directly and indirectly, placing public health at risk, (d) many intervention strategies have been developed to control and/or reduce the concentration of V. parahaemolyticus in oysters to acceptable levels, but most of them are mainly focused on the downstream steps of the oyster supply chain, and (c) although available regulation and/or guidelines governing the safety of oyster consumption are mostly available in developed countries, limited food safety information is available in developing countries. The information provided in this review may serve as an early warning for managing the future effects of climate change on the safety of oyster consumption.     

High pressure processing of shellfish: A review of microbiological and other quality aspects

Many commercially important shellfish are filter feeders and, as a consequence, concentrate microbes from the surrounding waters. Shellfish may be relayed or depurated to reduce the level of microbial contamination, but the efficiency of these purification practices, particularly in relation to viruses and indigenous marine bacteria, is questionable. Therefore additional processing is necessary to ensure the safety of shellfish for human consumption. In recent years high pressure (HP) processing has been investigated as an alternative method for food preservation. HP technology allows inactivation of microorganisms while maintaining sensory and nutritional properties of foods. Currently, HP processing has several commercial food applications, including oysters. As well as enhancing safety and extending shelf-life, HP treatment has the additional advantage of shucking or opening shellfish, making this technology particularly beneficial to the shellfish processing industry and consumers alike.Industrial relevanceHigh pressure (HP) processing is increasingly being used in the commercial processing of oysters, due to its minimal effects on sensory and nutritional quality, the opening or shucking of oysters during treatment, and the reduction of levels of Vibrio vulnificus, a pathogen of concern particularly in the US. However, little is known of the efficacy of HP treatment in reducing other pathogens in shellfish such as human enteric viruses, which are the predominant cause of shellfish-borne disease. This article reviews the inactivation of microorganisms of importance to shellfish, particularly viruses, the commercial HP processing of oysters and the advantages of HP technology as they pertain to the seafood industry.