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。     

上海市市售带壳牡蛎微生物污染状况调查

目的了解上海市市售带壳牡蛎的微生物污染状况。方法2005年4月至2006年3月在上海市水产品批发市场每月2次采集牡蛎样品,共抽取了24份。按照GB/T4789—2003《食品卫生微生物学检验》中检验规程进行微生物检测。结果所有抽检样品均不合格,污染最严重的是大肠菌群,所检样品均超标,最高污染量达1·4×106MPN/g;其次是菌落总数,超标率达41·7%,最高污染量达2·6×107CFU/g;副溶血性弧菌污染也比较严重,检出率达29·2%,污染量在3·0×102~7·4×102MPN/100g;单核细胞增生李斯特菌未检出。结论上海市市售带壳牡蛎的微生物污染状况不容乐观,已对消费者健康构成了潜在威胁。     

陕西省某活鸡屠宰场肉鸡相关样品中沙门菌定量检测及污染分析

目的了解肉鸡屠宰加工中不同时间和环节沙门菌的污染情况,分析污染关键点。方法 2016年11月至2017年11月从陕西省某活鸡屠宰场不同环节定期采集活鸡肛拭子标本、整鸡胴体和鸡肉样品,使用最大可能数(MPN)法对沙门菌进行定量检测,同时分离菌株;采用聚合酶链式反应(PCR)技术对沙门菌进行鉴定,同时结合血清凝集技术对沙门菌鉴定结果进行确认。结果采集的284份样品中有67份检出沙门菌,检出率为23. 6%,平均MPN值为0. 051 6 MPN/g。2017年7月采集的样品沙门菌污染最为严重,检出率为37. 8%(14/37),平均MPN值为0. 064 7 MPN/g; 2016年11月检出率最低,为13. 9%(5/36),平均MPN值为0. 043 6 MPN/g。不同屠宰环节中,浸烫褪毛后整鸡胴体样品中沙门菌检出率最高(43. 3%,26/60),平均MPN值为0. 060 5 MPN/g;分割后冷冻前鸡胸脯肉样品中沙门菌检出率最低(18. 3%,11/60),平均MPN值为0. 036 8 MPN/g,略高于储存配送过程整鸡胴体/鸡胸脯肉样品中沙门菌的污染水平(0. 035 8 MPN/g)。结论活鸡屠宰过程沙门菌的检出率与MPN值具有较强的季节性,在不同屠宰加工环节存在纵向和交叉污染,应对活鸡屠宰加工过程沙门菌污染严重的环节进行重点控制。     

福建省零售生食牡蛎中副溶血性弧菌的定量危险性评估

为模拟生牡蛎消费引起副溶血性弧菌(VP)疾病的危险性,在福建省开展了定量微生物危险性评估。评估遵循广泛认可的危险性评估程序,涉及危害识别、危害特征描述、暴露评估和危险性特征描述。结合暴露评估模块的结果与贝塔-泊松剂量反应模型,推测由消费VP污染的生牡蛎导致疾病发生的危险性分别是6.9×10-7(冬)、1.7×10-5(春)、5.9×10-5(夏)、4.6×10-6(秋)。敏感性分析结果表明,零售期间牡蛎的未冷藏时间、零售带壳牡蛎体VP密度的对数值、冷却持续时间和气温等因素,与疾病发生的危险性显著相关。采取缩短零售期间牡蛎的未冷藏时间、快速冷却、微热处理和冷冻贮存等控制措施,能够明显降低疾病发生的人数。该研究为我国制定减少VP对公众健康影响的政策提供了理论依据。     

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。     

东南沿海地区零售海产品中创伤弧菌的监测

目的了解我国东南沿海地区零售海产品中创伤弧菌的污染状况。方法采用3%氯化钠碱性蛋白胨水增菌,改良纤维二糖-多粘菌素B-多粘菌素E(mCPC)琼脂和纤维二糖-多粘菌素E(CC)琼脂分离海产品试样中的创伤弧菌,可疑菌落的鉴定采用生化试验或PCR法。采用最可能数(MPN)法和PCR法检测牡蛎试样的创伤弧菌污染水平。结果天然污染海产品试样创伤弧菌的检出率为19.8%(20/101)。牡蛎试样中45%(55/122)创伤弧菌密度低于3MPN/g的最低检出限。牡蛎中创伤弧菌的污染水平存在季节性差异。结论未来应加强对我国海产品中创伤弧菌污染状况的监测,尤其是在温暖的季节。     

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。     

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。     

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。     

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.     

Populations of Vibrio parahaemolyticus in retail oysters from Florida using two methods

Vibrio parahaemolyticus is a naturally occurring estuarine bacterium that is often associated with gastroenteritis in humans following consumption of raw molluscan shellfish. A number of studies have investigated the environmental distribution of V. parahaemolyticus, but little is known about the levels of this organism during distribution of oysters or at the point of consumption. Duplicate samples of shellstock oysters were collected monthly (September 1997 to May 1998) from the same four restaurants and three wholesale seafood markets in the Gainesville, Fla. area and analyzed for total V. parahaemolyticus densities using two methods: a standard MPN method (BAM-MPN) and a new direct plating procedure (direct-VPAP). Both methods employed an alkaline phosphatase-labeled DNA probe (VPAP) targeting the species-specific thermolabile hemolysin (tlh) gene to confirm suspect colonies as V. parahaemolyticus. The highest monthly geometric mean V. parahaemolyticus density was observed in October of 1997 (approximately 3,000/g) with similarly high values during September and November of 1997. From December 1997 to May 1998 mean densities were generally less than 100/g, falling to approximately 10/g in February and March. A strong correlation (r = 0.78) between the direct-VPAP and BAM-MPN methods for determining V. parahaemolyticus densities in market-level oysters was observed. The direct-VPAP method was more rapid and precise while the BAM-MPN was more sensitive and may better recover stressed cells. The utilization of the VPAP probe for identification of V. parahaemolyticus sharply reduced the labor for either method compared to biochemical identification techniques used in earlier V. parahaemolyticus surveys.     

Occurrence of Vibrio parahaemolyticus in Two Oregon Oyster-growing Bays

ABSTRACT: Occurrence of Vibrio parahaemolyticus in 2 Oregon oyster-growing areas (Yaquina andTillamook Bays) was studied from November 2002 to October 2003. Vibrio parahaemolyticus was detected in 15.0% of oyster, 20.0% of seawater, and 47.5% of sediment samples with very low levels of pathogenic strains being detected in oysters (≤3.6 most probable number [MPN] /g). The densities of total and pathogenic V. parahaemolyticus were higher in sediment (≤1100 and ≤43 MPN/g) than in seawater (≤15 and ≤3.6 MPN/100 mL) or oyster (≤43 and ≤3.6 MPN/ g). Densities of V. parahaemolyticus in both bays were positively correlated to water temperatures ( P < 0.01), with higher densities in samples being detected in summer, especially July and August. There was no correlation between the densities of V. parahaemolyticus and water salinity or the densities of V. parahaemolyticus and bacterial populations in seawater. Freshly harvested oysters should be kept at refrigeration temperatures to prevent rapid growth of pathogenic V. parahaemolyticus in contaminated oysters.     

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.     

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.     

上海市零售梭子蟹中副溶血性弧菌的污染状况及风险评估

目的 研究上海市场零售梭子蟹中副溶血性弧菌的污染状况,并对副溶血性弧菌可能引发的公共卫生风险进行初步评估.方法 采用最可能数(MPN)法检测上海和宁波市售梭子蟹中副溶血性弧菌的带菌量,采用膳食回顾性调查方法对上海居民梭子蟹消费量进行调查,采用Risk Ranger软件对梭子蟹中副溶血性孤菌进行丰定量风险评估.结果 上海和宁波市零售梭子蟹中副溶血性弧菌阳性率分别为60.5%和13.3%,阳性试样平均菌量分别为0.75 MPN/g和5.50 MPN/g.膳食调查结果显示,上海居民梭子蟹人均日摄入量为2.95 g/d.风险评估结果显示,风险评分为36分,梭子蟹食用者每人每天的发病概率为8.22×10-9,每年上海地区预期发生42.5例副溶血性弧菌感染病例.通过改变评估模型中某些风险的选择,如采取良好的控制措施,风险可降低至原来的1/10.结论 上海市售梭子蟹副溶血性弧菌污染状况较为严重,可能对人群健康造成较高的风险,未来有必要进一步加强监测.     

Growth and survival of Vibrio parahaemolyticus in postharvest American oysters

Oysters at the retail stage of distribution generally contain greater densities of Vibrio parahaemolyticus than do oysters at harvest. The objective of this study was to determine the effects of postharvest storage at 26 and 3 degrees C on the growth and survival of naturally occurring V. parahaemolyticus in shellstock American oysters (Crassostrea virginica). Oysters were collected monthly from May 1998 through April 1999 from Mobile Bay, Alabama, and their V. parahaemolyticus densities were determined after 0, 5, 10, and 24 h of postharvest storage at 26 degrees C. After 24 h of storage at 26 degrees C, oysters were transferred to a refrigerator at 3 degrees C and analyzed 14 to 17 days later. V. parahaemolyticus numbers were determined by a direct plating method involving an alkaline-phosphatase-labeled DNA probe that targets the species-specific thermolabile hemolysin gene (tlh-AP) to identify suspect isolates. From April to December, when water temperatures at harvest were >20 degrees C, the geometric mean harvest density of V. parahaemolyticus was 130 CFU/g. When water temperatures were <20 degrees C, the geometric mean harvest density was 15 CFU/g. After harvest, V. parahaemolyticus multiplied rapidly in live oysters held at 26 degrees C, showing a 50-fold increase (1.7 log CFU/g) at 10 h and a 790-fold increase (2.9 log CFU/g) at 24 h (April through December). Average V. parahaemolyticus numbers showed a sixfold decrease (0.8 log CFU/g) after approximately 14 days of refrigeration. These results indicate that V. parahaemolyticus can grow rapidly in unrefrigerated oysters.