三亚市售海白中副溶血性弧菌半定量风险评估

对海白中副溶血性弧菌(Vibrio parahaemolyticus,简称Vp)做半定量风险评估,包括危害识别、污染检测、居民膳食调查及Risk ranger半定量风险评估。膳食调查采用食物频率法,以问卷的方式得到三亚居民食用海白的比例为80%及食用频率为1月1次;污染检测主要3月、4月份期间从三亚第一市场和荔枝沟市场取样,采用GB/T 4789.7-2008法对样品进行检测,检出率为100%并得到两个市场的平均污染量分别为1.65 log10MPN/g,2.63log10MPN/g;最终用Risk ranger得到三亚市售海白的副溶血性弧菌膳食风险为高风险(风险指数为51),三亚海白消费者感染副溶血性弧菌胃肠炎的概率为2.96×10-6(人·d)-1;每年消费污染了副溶血性弧菌的海白所引发的病例数约为740.5人。     

淄博市市售生食贝类中副溶血性弧菌污染水平调查

为了解山东省淄博市市售生食贝类中副溶血性弧菌的污染状况和毒力基因的携带情况,分别于2019年8、9、10月从不同区县的零售市场采集牡蛎和毛蚶样品共40份,运用副溶血性弧菌MPN定量法和多重荧光MPN-PCR法对副溶血性弧菌的污染状况和毒力基因的携带情况进行调查。结果显示,副溶血性弧菌(tlh基因)检出率为47.50%(19/40),其中污染量≥110MPN/g的样品占26.00%(5/19);毒力基因(tdh、trh)阳性15份,携带率37.50%(15/40),其中牡蛎携带率为45.00%(9/20),毛蚶携带率为30.00%(6/20)。故得出结论,淄博市市售牡蛎和毛蚶中副溶血性弧菌的污染水平较高,存在一定的食品安全风险。     

2008—2010年上海市夏秋季市售海产品中副溶血性弧菌污染监测结果分析

目的 了解上海市夏秋季节市售海产品中副溶血性弧菌的污染水平和特征.方法 2008-2010年5-10月,采用GB/T 4789.7-2008《食品卫生微生物学检验副溶血性孤菌检验》方法,对上海市批发市场、集贸市场、卖场超市和餐饮单位等污染物监测点的市售海产品进行副溶血性弧菌的定性和定量检测.结果 共监测市售海产品941件,副溶血性弧菌总体检出率为13.2％,不同种类、不同监测月份和不同采样地点的海产品,其副溶血性弧菌检出率和样品几何平均浓度总体上差异有统计学意义(P＜0.05).其中,海产虾类副溶血性弧菌检出率(25.0％)和样品几何平均浓度(5.0 MPN/g)显著高于其他类海产品(P＜0.05);8月份海产品副溶血性弧菌检出率(27.4％)和样品几何平均浓度(3.3 MPN/g)显著高于其他监测月份(P＜0.05);集贸市场副溶血性弧菌检出率(28.5％)和批发市场样品几何平均浓度(3.9 MPN/g)显著高于其他采样地点(P＜0.05).结论 上海市市售海产品中副溶血性孤菌的污聚率较高,应进一步开展海产品中副溶血性弧菌的风险监测和评估,并针对副溶血性弧菌污染的高风险环节开展监管.     

生食动物性水产品中副溶血性弧菌和创伤弧菌污染状况分析

目的 了解生食动物性水产品中副溶血性弧菌和创伤弧菌污染状况。方法 采用随机抽样方法,在我国13个地区的餐饮店、零售店和批发市场采集生食动物性水产品,共计2 980份,对样品进行副溶血性弧菌和创伤弧菌检测。结果 生食动物性水产品中副溶血性弧菌检出率为14.7%(437/2 980),污染水平＞100 MPN/g的样品比例为2.9%(83/2 909);创伤弧菌检出率为3.5%(104/2 980)。采样于批发市场的样品中副溶血性弧菌检出率、污染水平＞100 MPN/g的样品比例和创伤弧菌检出率均高于餐饮店和零售店。第三季度副溶血性弧菌检出率、污染水平＞100 MPN/g的样品比例和创伤弧菌检出率最高。造成污染的主要原因包括原产地污染,储存不当及加工过程交叉污染。结论 生食动物性水产品中存在副溶血性弧菌和创伤弧菌的污染,其健康风险应引起关注,本次污染状况分析可为标准制修订提供理论依据。     

上海市水产品副溶血性弧菌的污染情况及毒力基因分布

副溶血性弧菌是引起沿海城市食源性疾病爆发的主要致病菌之一。为了调查2015年7月—2016年6月上海市内水产品中副溶血性弧菌的污染情况及毒力菌株的分布,根据GB/T 4789. 2013《食品卫生微生物学检验副溶血性弧菌检验》方法结合聚合酶链式反应及环介导等温扩增技术,从206份水产样品中分离出201株疑似副溶血性弧菌,挑取疑似菌株接种至科玛嘉弧菌显色培养基并获取最大可能数,提取菌株基因组DNA对其进行毒力基因鉴定。从上海芦潮港码头收集71份样品中,有59份样品中分离出副溶血性弧菌,污染量为88.36 MPN/hg。上海市水产市场采样副溶血性弧菌污染率高达91.11%,污染量为1 361.76 MPN/hg。其中,tdh、trh阳性菌株污染率分别为1.6%、1.0%,均分离自虾中。研究表明,上海市售水产品中虾类是总副溶血性弧菌及致病性副溶血性弧菌污染率最高的水产品种类。水产品市场相对于自然环境而言具有更高的污染率和污染量,水产品捕捞后的一系列操作增加了副溶血性弧菌的污染。这些结果为上海市食源性疾病的防控提供了依据。     

南宁市对虾中副溶血性弧菌的污染情况调查及对虾消费情况分析

目的:了解南宁市市售对虾中副溶血性弧菌的污染状况及对虾消费情况,为建立副溶血性弧菌风险评估及安全预警提供科学依据。方法:从南宁市各大超市、海鲜市场、菜市场和酒店采集鲜活对虾,根据GB 4789.7-2013标准进行对虾中副溶血性弧菌的分离鉴定;采用问卷调查的形式了解南宁市居民消费对虾的情况。结果:发现91份对虾样品中,检测出副溶血性弧菌的样品数为50份,总检出率为54.9%。对虾生食的比例平均值为5.57%,居民平均消费对虾的数量为13只/天。     

北京市水产品污染与感染病例中副溶血性弧菌血清型和毒力基因型的比较研究

目的对北京市夏季市售水产品污染与感染病例中副溶血性弧菌血清型和毒力基因型进行比较研究,为评估食品安全风险监测的目的与意义提供思路,为北京市水产品副溶血性弧菌污染与临床感染病例的关联性研究提供技术支持。方法对采集的水产品样品和哨点医院腹泻患者粪便样本进行副溶血性弧菌的分离鉴定,采用血清玻片凝集法对分离出的副溶血性弧菌进行血清分型,PCR方法检测菌株的tlh、tdh、trh基因。结果 2014年7~9月共采集水产品样品164份,检出副溶血性弧菌80份,总污染率为48.78%;其中淡水产品污染率为38.78%(19/49),平均菌量浓度为66.63 MPN/g;海水产品污染率为53.04%(61/115),平均菌量浓度为38.14 MPN/g。80株副溶血性弧菌分属于9个血清群,其中O2群28株,占35.00%,O1群11株,占13.75%,O5群10株,占12.50%。80株菌tlh基因均为阳性,只有1株菌携带tdh毒力基因,所有菌株trh毒力基因均为阴性。哨点医院腹泻病人粪便样本中分离鉴定副溶血性弧菌21株,血清型O3∶K6占61.90%(13/21),O4:K8占28.57%(6/21);毒力基因型tdh(+)/trh(-)占95.24%(20/21),tdh(-)/trh(-)占4.76%(1/21)。结论来源于食品样品的副溶血性弧菌绝大部分不具备致病性,而导致消费者腹泻的副溶血性弧菌绝大部分携带致病性毒力基因,表明目前的食品安全风险监测结果不能作为评估副溶血弧菌导致的食源性疾病暴发和散发的依据。     

茶多酚对泥蚶中副溶血性弧菌的抑制效果

为了降低食用泥蚶引起的副溶血性弧菌食源性疾病的风险,研究了天然抑菌剂茶多酚对泥蚶中副溶血性弧菌的抑制作用并做出感官评价。副溶血性弧菌的定量采用三管最大可能值法(MPN法)。研究结果表明,茶多酚对副溶血性弧菌的最低抑制浓度(MIC)为0.5 g/L;带菌泥蚶经0.5 g/L的茶多酚处理后于0℃,5℃和10℃下贮藏60h,副溶血性弧菌数量分别下降了2.75 Log10 MPN/g,2.59 Log10 MPN/g和2.15 Log10 MPN/g,杀菌率均在99%以上。经感官评定小组评定,茶多酚浓度为0.5 g/L时,对泥蚶的色泽、气味、口感等感官特性不产生负面影响。茶多酚是一种理想的天然抑菌剂。     

湛江两近海域文蛤和翡翠贻贝副溶血性弧菌污染情况分析

目的:对湛江东海岛和南三岛两近海域文蛤和翡翠贻贝副溶血性弧菌的污染进行检测和评价。方法:在5～11月,采用SN0173-92出口食品副溶血性弧菌检验方法,分别对文蛤和翡翠贻贝样品进行副溶血性弧菌生化反应初筛和MPN值测定。结果:每克文蛤样品中副溶血性弧菌的MPN值为<3～93,其中在6～7月副溶血性弧菌MPN值最高,为93。每克翡翠贻贝样品中副溶血性弧菌的MPN值<3～23,其中在6～7月副溶血性弧菌MPN值最高,为23。结论:湛江东海岛和南三岛附近海域每克文蛤和翡翠贻贝样品中副溶血性弧菌MPN最高值≤100,未超过相关国际标准。     

2003年中国部分沿海地区零售海产品中副溶血性弧菌污染状况的主动监测

国家食源性疾病监测网发现，我国近年来副溶血性弧菌中毒呈显上升趋势。为进一步了解零售海产品中副溶血性弧菌(VP)的污染情况，2003年9-12月在我国沿海4个省份(浙江、江苏、广东、福建)进行监测，试样分别从水产品批发市场、零售市场和饭店采集，共采集海产品236份，其中甲壳类69份、贝类116份、鱼类51份。采用Vitek鉴定系统和最可能数(MPN)法进行副溶血性弧菌的定性和定量分析。结果显示，38．6％的海产品检出VP，浙江省试样的VP阳性率最高。甲壳类、贝类和鱼类试样VP阳性率分别为49．3％、37．9％和25.5％；阳性试样几何平均分布浓度依次为171．4、76．9和50．7MPN／100g。监测结果表明，我国零售海产品中副溶血性弧菌的污染率较高，必须持续地进行食品中VP的主动监测和污染控制。     

温暖月份零售带壳牡蛎中副溶血性弧菌的定量研究

为了解温暖月份零售带壳牡蛎中副溶血性弧菌 (VP)的污染情况 ,2 0 0 3年 4～ 8月在福建省福州和厦门两地共收集带壳牡蛎 113份 ,样品分别来自水产品批发市场 (18% ) ,零售市场 (4 6 % )和饭店 (36 % )。采用Vitek鉴定系统和最可能数法进行VP的定量分析。结果显示 ,带壳牡蛎中VP密度的几何均数为 6 0MPN 10 0g ,4 1 6 %的样品VP密度低于 30MPN 10 0g的最低检出限 ,仅厦门2个样品菌量超过 2 4 0 0 0MPN 10 0g。两个地区、不同采样点和不同月份之间样品VP密度的几何均数差别均有统计学意义 (P <0 0 1)。厦门样品污染菌量高于福州 ;批发市场样品菌量最高 ;5月份样品菌量最高 ,为 14 9MPN 10 0g ,而 6～ 8月样品菌量约为 4 0MPN 10 0g。零售环节带壳牡蛎VP的检出率较高。未来应加强对生食海产品中VP污染状况的监测。     

福建省带壳牡蛎中副溶血性弧菌的市场调查

为了解零售带壳牡蛎中副溶血性弧菌(VP)的污染情况，2003年4月～2004年3月每月在福建省福州和厦门两地收集带壳牡蛎，样品共252份，分别来自水产品批发市场(11％)、零售市场(50％)和饭店(39％)。采用Vitek鉴定系统和最可能数(MPN)法进行VP的定性和定量分析。结果显示，带壳牡蛎VP几何平均密度为46MPN，100g，46％的试样VP密度低于30MPN/100g的最低检出限，仅厦门2个试样菌量超过104MPIN/100g。两个地区、不同采样点和不同季节之间试样vP平均密度差别均有显性。厦门试样菌量高于福州；批发市场试样菌量最高；春季试样菌量(93MPN/100g)高于其它季节(约为40MPN/100g)。研究结果可以用于估计生食牡蛎人群VP的暴露量。     

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.     

上海市生食三文鱼中副溶血性弧菌污染的风险分析

为了解上海市生食水产品的安全状况,为食品安全分析和监管提供理论依据,研究上海市生食三文鱼为代表的生食水产品中副溶血性弧菌的污染情况,对上海市3 个批发市场进行监测分析,在2013年基于GB/T 4789.7-2008《食品卫生微生物学检验：副溶血性弧菌检验》方法共检测了90 个样品,副溶血性弧菌检出率为20.00%,其中致病性副溶血性弧菌（tdh＋）的检出率为1.10%,使用MEGA软件根据16S rDNA序列开展了菌株多样性分析;根据调查的副溶血性弧菌在生食三文鱼中的污染情况,使用@Risk 5.5 软件分析了生食三文鱼可能引发副溶血性弧菌食物中毒的风险,经Beta-Poisson剂量反应模型、蒙特卡洛模拟等分析后表明,生食三文鱼患病的概率为2.02×10－6,即每百万人有2 人发病;根据上海市三文鱼的平均消费总量,上海市居民每年生食三文鱼因副溶血性弧菌污染可能的患病人数为111 人。上海市居民生食三文鱼由副溶血性弧菌污染引起疾病的风险较小,控制副溶血性弧菌的污染量和每次三文鱼的消费量是减少风险的关键。     

Levels of Vibrio parahaemolyticus and thermostable direct hemolysin gene-positive organisms in retail seafood determined by the most probable number-polymerase chain reaction (MPN-PCR) method

The incidence and levels of Vibrio parahaemolyticus and thermostable direct hemolysin gene (tdh)-positive organisms in retail seafood were determined. The most probable number-polymerase chain reaction (MPN-PCR) method using a PCR procedure targeting the species-specific thermolabile hemolysin gene (tlh) and tdh was used to determine the levels of V. parahaemolyticus and tdh-positive organisms, respectively. In seafood for raw consumption, V. parahaemolyticus was found in four (13.3%) of 30 fish samples, 11 (55.0%) of 20 crustacean samples, and 29 (96.7%) of 30 mollusc samples. Levels of V. parahaemolyticus were below 10(4) MPN/100 g in all fish and crustacean samples tested. However, they were above 10(4) MPN/100 g in 11 (36.7%) of the 30 mollusc samples. In all seafood for raw consumption, the level of tdh-positive organisms was below the limit of detection (< 30 MPN/100 g). In seafood for cooking, V. parahaemolyticus was found in 15 (75.0%) of 20 fish samples, nine (45.0%) of 20 crustacean sample, and 20 (100%) of 20 mollusc samples. Levels of V. parahaemolyticus were above 10(4) MPN/100 g in only three (15.0%) and one (5.0%) of the 20 fish and 20 crustacean samples, respectively. However, they were above 10(4) MPN/100 g in 18 (90.0%) of the 20 mollusc samples. In seven (35.0%) of the 20 mollusc samples, tdh-positive organisms were found and their levels ranged from 3.6x10 to 1.1 x 103 MPN/100 g. From four of seven tdhpositive samples, tdh-positive V. parahaemolyticus was isolated.     

Occurrence and density of vibrio parahaemolyticus in live edible crustaceans from markets in China

Vibrio parahaemolyticus is a marine bacterium causing foodborne disease. Occurrence of the bacterium was investigated in six species of edible crustaceans available from markets in mainland China. The bacterium was detected in 22 of 45 whole-body, shell, and feces samples, including mitten crabs, which are supposed to be produced in freshwater ponds. The mean densities ranged from 2.8 log CFU/g in mitten crabs (Eriocheir sinensis) to 5.1 CFU/g in giant tiger prawns (Penaeus monodon). In hemolymph and muscle samples collected axenically, V. parahaemolyticus was detected in all of the prawns at a mean density of 2.6 log most probable number (MPN)/g, in two of five striped stone crabs (Charybdis feriatus) at a mean density of 1.1 log MPN/ml, and two of five mangrove mud crabs (Scylla serrata) at a mean density of 1.3 log MPN/ml. When six mitten crabs were collected from two freshwater ponds in China and were examined, V. parahaemolyticus was not detected. It seemed that cross-contamination occurred among live crustaceans at the markets. The results suggest that proper handling, storage, and cooking of these crustaceans will be necessary to lessen the risk of foodborne illness from V. parahaemolyticus.     

Bactericidal effects of wine on Vibrio parahaemolyticus in oysters

The bactericidal effects of wines on Vibrio parahaemolyticus in oysters were studied to evaluate potential inactivation of V. parahaemolyticus in contaminated oysters by wine consumption. Shucked whole oyster and oyster meat homogenate were inoculated with V. parahaemolyticus and mixed with red or white wine. Survivals of V. parahaemolyticus in inoculated oysters were determined at 7 and 25 degrees C. Populations of V. parahaemolyticus in inoculated whole oysters (5.52 log most probable number [MPN] per g) decreased slightly to 4.90 log MPN/g (a 0.62-log reduction) after 24 h at 7 degrees C but increased to 7.37 log MPN/g over the same period at 25 degrees C. However, the populations in wine-treated whole oysters decreased by >1.7 and >1.9 log MPN/g after 24 h at 7 and 25 degrees C, respectively. Both red and white wines were more effective in inactivating V. parahaemolyticus in oyster meat homogenate than in whole oyster. Populations of V. parahaemolyticus in oyster meat homogenate (7.8 x 10(3) MPN/g) decreased rapidly to nondetectable levels (< 3 MPN/g) after 30 min of mixing with wine at 25 degrees C (a 3.89-log MPN/g reduction). These results suggest that chewing oysters before swallowing when eating raw oysters may result in greater inactivation of V. parahaemolyticus if wine is consumed. More studies are needed to determine the bactericidal effects of wine on V. parahaemolyticus in the complicated stomach environment.     

Evaluation of most-probable-number-PCR method with internal amplification control for the counting of total and pathogenic Vibrio parahaemolyticus in frozen shrimps

The most-probable-number (MPN) method is often time-consuming for the isolation, detection, and quantification of Vibrio parahaemolyticus from natural sources. MPN counting of V. parahaemolyticus bacteria usually involves the isolation of typical V. parahaemolyticus colonies on selective medium, with subsequent confirmation by biochemical identification. In this study, we evaluated the use of a PCR on MPN enrichment cultures (MPN-PCR) for the direct detection of total and pathogenic V. parahaemolyticus cells in frozen shrimp. This reaction targeted the R72H, tdh, and trh sequences. An internal amplification control was added to the samples before R72H amplification. There was an excellent correlation between the results of the two methods for artificially inoculated and natural shrimp samples. Of 36 natural samples, 28 tested positive for the presence of V. parahaemolyticus, with an MPN value of 2 × 10(-1) to 9.2 × 10(1) per g. No pathogenic V. parahaemolyticus cells were detected. The test had a detection limit of one V. parahaemolyticus organism per g and was completed within two working days. These results support the use of the combination of PCR with MPN for the detection of total or potentially pathogenic V. parahaemolyticus cells in frozen shrimp.