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。     

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。     

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。     

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。     

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.     

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。     

短期净养系统对带壳牡蛎品质的改善作用

为解决牡蛎肉质洁净度难以保证的问题,设计了一种一体式短期净养系统作为牡蛎进一步加工前的暂养技术,包括净养池、过滤装置、蛋白泡沫分离装置、紫外杀菌装置及制冷装置等。分别针对复原海水盐度(0.00、0.85、1.70、2.55、3.40 g/100 mL)、水体循环、贝水质量比(1∶400～1∶22.5)3个因素对带壳牡蛎品质的影响进行单因素试验。结果表明:当复原海水盐度为2.55 g/100 mL,同时进行伴有过滤、蛋白泡沫分离、通氧等装置的水体循环时,净养24 h后带壳牡蛎存活率为98.3%,肉质盐度为2.55 g/100 mL,洁净度最高;此外,贝水质量比为1∶30时,牡蛎存活率和肉质洁净度能够得到保证,水体菌落总数上升较贝水质量比为1∶22.5减慢72.6%;进一步将短期净养系统与置于装有蓄冷冰袋的普通泡沫箱暂存相比,带壳牡蛎的存活期延长87.5%,贮藏5 d时菌落总数降低约0.4(lg(CFU/g)),说明该短期净养系统能有效延长带壳牡蛎进一步加工前的暂存保质期。     

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.     

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。     

净化后的褶牡蛎细菌菌相分析

采集浙江省宁海县西店养殖的褶牡蛎去壳取肉,经过臭氧水杀菌净化处理后,对牡蛎体内细菌菌落总数、大肠菌群和细菌菌群组成进行了分析研究。牡蛎体内需氧平板菌落数为4.3×103cfu/g,大肠菌群数为157MPN/100g。从牡蛎体内共分离出细菌52株,通过鉴定,96.2%是革兰阴性杆菌,3.8%是革兰阳性杆菌,本研究没有分离到球菌。优势菌为弧菌属(Vibrio)28株,其次是黄杆菌属(Flavobocterium)6株、假单胞菌属(Pseudomonas)5株、无色菌属(Achromobacter)5株、气单胞菌属(Aeromonas)3株,此外还有肠杆菌科(Enterobacteriaceae)2株、杆状菌属(Bacillus)2株,另有1株细菌在传代中失去了生长能力。     

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.     

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。     

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.     

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.     

Oyster-to-oyster variability in levels of Vibrio parahaemolyticus

This study examined the variability in the levels of total and pathogenic Vibrio parahaemolyticus in individual oysters. Twenty oysters were collected on three occasions (in June, July, and September 2001) from a site near Mobile Bay, Ala. Ten of these oysters were tested immediately, and 10 were tested after 24 h of storage at 26 degrees C. Levels of total and pathogenic V. parahaemolyticus were determined by alkaline phosphatase-labeled DNA probe procedures targeting the thermolabile hemolysin and thermostable direct hemolysin genes, respectively. Similar V. parahaemolyticus levels (200 to 2,000 CFU/g) were found in nearly 90% of the oysters (for all sampling occasions) prior to storage. The log-transformed densities (means +/- standard deviations) of V. parahaemolyticus in oysters immediately after harvest were 2.90 +/- 0.91, 2.88 +/- 0.36, and 2.47 +/- 0.26 log10 CFU/g for June, July, and September, respectively. After storage for 24 h at 26 degrees C, the mean V. parahaemolyticus densities increased approximately 13- to 26-fold. Before storage, pathogenic V. parahaemolyticus was detected in 40% (10 to 20 CFU/g) of the oysters collected in June and July but was not detected in any oysters collected in September. After storage, pathogenic V. parahaemolyticus was detected in some oysters at levels of > 100 CFU/g. These data should aid in the development of sampling protocols for oyster monitoring programs and in the determination of exposure distributions associated with raw oyster consumption.