育肥牛场致病性大肠埃希菌及沙门菌的分离鉴定

采用常规肠道杆菌分离技术,对青海省湟中县某育肥牛场的60份和互助县某育肥牛场的14份粪样及环境样品进行致病性大肠埃希菌和沙门菌的分离与鉴定,结果显示湟中县某育肥牛场致病性大肠埃希菌检出率为6.7％,沙门菌检出率为15.0％;互助县某育肥牛场粪样致病性大肠埃希菌检出率为21.4％,沙门菌检出率为7.1％.药敏试验结果显示...     

银黑狐空肠弯曲菌和致病病性大肠埃希氏菌带菌状况的研究

对２２６只银黑狐粪便拭子进行空肠弯曲菌检查，鞭检出带菌狐１４只，其中成年公狐，成年母狐，母仔狐和公仔狐分别检出３，８，２和１只，检出率依次为７．５％，６．６％，５．０％和４．０％平均检出率为６．２％。对１８９只银黑狐粪例进行致病性大肠埃工菌检查，检出带菌狐３１只，其中成年公狐、丰年母狐、母仔狐（５１只）和公仔狐（３只），分别检出４，１８，８和１只，检出率依闪为３０．８％，１４．８％，１５．７％和３     

西宁野生动物园灵长类动物粪便中小肠结肠炎耶尔森氏菌的检测

应用细菌常规病原分离鉴定技术对西宁市野生动物园无菌采集的部分灵长类动物粪便进行小肠结肠炎耶尔森氏菌株的带菌率及其耐药率进行了检测.结果显示,在34份样品中共检出5株小肠结肠耶尔森氏菌,检出率为14.7％ (5/34);药敏试验结果表明,5株分离菌株对36种药敏纸片的平均耐药率为24.44％;致病性试验表明,4株分离菌对小鼠具有致死性效应,1株分离菌对小鼠有一定致病性.     

鸡致病性大肠杆菌菌毛分型的初步研究

以鸡致病性大肠杆菌F1菌毛特异性单抗1F4－1、2C3、3H3和F11菌毛特异性单抗FA1、FB11作为检测试剂，将111株已知O抗原的鸡致病性大肠杆菌经MD液体培养基连续传代培养后，通过直接玻板凝集法对各菌毛进行初步分型。结果发现F1、F11菌毛与O78、O1及O2三种优势O抗原型菌株之间存在较为明显的相关性，即致病性大肠杆菌主要集中在上。F1、F11菌毛在这三种O抗原型上的总检出率分别为95．6％、75．4％及73．3％。另外，在所检测的111株鸡致病性大肠杆菌中，只表达F1菌毛的大肠杆菌占菌杆总数的33．3％。只表达F11菌毛的大肠杆菌占菌株总数的8．1％，两者都表达的占菌株总数的36％，F1、F11菌毛的总检出率为78．3％。     

奶牛隐性乳腺炎病原菌的分离鉴定

用兰州乳房炎试验(LMT)对来自西安市某个牛场的76份奶样作隐性乳房炎检验，并对乳样进行细菌分离鉴定。结果表明，该牛场的隐性乳房炎检出率为33％，阳性乳样中细菌检出率为93．4％。阴性乳样中病原菌检出率为28％，在检出的94株细菌中，共有5种菌22株是与乳房炎有关的病原菌。其中葡萄球菌和链球菌占病原菌总数的90％以上。是引起该场奶牛隐性乳房炎的主要病原菌。     

鸵鸟大肠埃希氏菌的分离鉴定

从某鸵鸟养殖场发病的幼鸵鸟肝脏和脾脏分离到一种细菌，通过培养特性，菌体形态，菌落形态，染色特性，生化试验等一系列的系统鉴定，确定为大肠埃希氏菌，动物致病性试验表明该菌对小白鼠和雏鸡有较强的致病性，药敏试验证明该菌对常见的抗生素不敏感，经血清型鉴定该菌血清型为O1。     

用生乳增菌法检测乳中葡萄球菌

用生乳增菌法和常规法检查６５份乳样中的葡萄球菌。生乳增菌法检出２１株，葡萄球菌检出率为３２．３１％，常规法检出８株，检出率１２．３１％，两种方法对葡萄球菌的检出率差异极显著（Ｐ〈０．０１）。     

鱼嗜水气单胞菌的分离鉴定

从陕西省甘养殖场送检的病鱼中分离到1株革兰氏阴性杆菌，培养特征观察笔生化特性分析结果证实为嗜水气单胞菌；小鼠和金鱼致病性试验证实分离菌具有致病性；药敏试验结果表明该菌对链毒素、万古霉素等高度敏感，对羧苄青霉素、氟哌酸等不敏感。用该菌制成灭活免疫金鱼保护力可达到80％以上。     

保定地区羊源致病性粪肠球菌的分离鉴定及其毒力性与耐药性分析

为探究保定地区羊源致病性粪肠球菌的致病性与耐药程度，采集30只同时发生肺炎与腹泻的病羊肝脏、肺脏等内脏器官，进行病理学观察及病原体的分离培养、溶血性鉴定、16S rRNA测序、致病性试验、药敏试验、毒力基因与耐药基因的检测。结果显示，共分离到17株致病性粪肠球菌，分离率为56.67%(17/30)。在这些菌株中，检测出6种毒力基因，其中ace基因的检出率最高，检出率为100.00%(17/17)。分离菌对阿奇霉素、红霉素、庆大霉素、克林霉素4种药物的耐药率较高，为100.00%(17/17);对氨苄西林、阿莫西林、青霉素G等3种β-内酰胺类药物的耐药率较低，分别为11.76%(2/17),11.76%(2/17)和0.00%(0/17)。耐药基因中，大环内酯类ermB基因检出率为100.00%(17/17),未检出β-内酰胺类基因TEM。本研究为羊源致病性粪肠球菌的诊断、治疗和预防提供了依据。     

合肥地区类志贺邻单胞菌的动物贮存宿主调查

为探讨合肥地区类志贺邻单胞菌的动物贮存宿主，采用二次增菌分离培养法，对１５种２２８份标本进行检测，结果１２处３６份样品检出此菌，总阳性检出率为１５．７９％，鱼类、观赏水禽、家鸭、猪和清摘工，阳性检出率分别为２３．２９％、１７．８６％、１３．０４％、６．６７％和１０％，表明该菌具有广泛动物贮存宿主，尤其是淡水鱼类，同时对３６株阳性菌的血清分群，其中１７株可分群，分群率为４７．２２％，共分布于７个血清型，人源与动物源共有的血清群是Ｏ１９     

苏州大型超市小水产品中副溶血性弧菌的污染调查及耐药性分析

目的了解副溶血性弧菌(Vibrio parahaemolyticus)在苏州市大型超市小水产品中的污染状况及耐药性,为建立副溶血性弧菌食物中毒预警系统提供科学依据。方法定期从苏州一大型超市抽取小水产品,根据GB/T4789.7-2008标准进行副溶血性弧菌的分离培养和鉴定,应用K-B法进行药敏实验。结果在144份样品中检出副溶血性弧菌56株,检出率38.9%。所有分离菌株对先锋必素、诺氟沙星和环丙沙星敏感;部分菌株对氨苄西林、阿莫西林、复方新诺明和头孢噻吩等具有较强的耐药性,耐药率分别为55.4%、42.9%、35.7%和32.1%;有9株菌株出现了多重耐药。结论苏州大型超市小水产品中副溶血性弧菌污染严重,应加强副溶血性弧菌食源性疾病预警,同时加强水产品抗生素使用的管理,防止其耐药菌株的传播。     

副溶血性弧菌耐药基因的研究进展

副溶血性弧菌为水产品主要致病菌之一,水产业和农业中抗生素的过量使用导致副溶血性弧菌对所推荐使用的抗生素产生了抗性,并且在副溶血性弧菌中已经发现了质粒和整合子等基因移动元件,更增加了与其他细菌发生耐药基因相互交换和重组的几率。对副溶血性弧菌的耐药研究现状、耐药机制和耐药基因的水平转移进行综述。     

副溶血弧菌SHJLA株的分离鉴定及致病性分析

副溶血性弧菌（Vibrio parahaemolyticus）是海洋环境中常见的食源性致病茵。本研究从对虾中分离出1株细菌SHJLA,在TCBS和弧菌显色培养基上分别显示典型的蓝绿色和紫红色的菌落,且其生理生化特性具有典型副溶血弧菌的特性。以SHJLA菌株的基因组为模板。检测副溶血弧菌种特异性基因（tlh、toxR、groEL）均为阳性,gyrB基因序列分析表明SHJLA与副溶血弧茵的亲缘关系最近,同源性达98％～100％;检测副溶血弧菌主要毒力相关基因tdh和T3SS2（VopC2和vcrD2）均为阳性。该菌的神奈川溶血实验为阳性,对小鼠的半数致死量（LD50）为4．8×10^8 cfu／mL。结合SHJLA菌株的形态、生理生化、种特异性基因的检测、gyrB基因序列分析、毒力相关基因的检测以及小鼠半数致死量的测定结果,确定SHJLA是一株携带毒力基因的副溶血弧菌。     

舟山沿海贝类产品中副溶血性弧菌的检测与毒力基因分析

为了解舟山沿海贝类产品中副溶血性弧菌污染情况,我们于2008年3个不同季节,从不同的农贸市场采集贝类标本,采用实时荧光定量PCR检测方法和常规培养方法,对舟山沿海贝类产品中的副溶血性弧菌进行了检测,同时对分离到的菌株进行血清学分型和毒力基因检测。结果60份贝类产品中,采用常规的分离培养方法,阳性率为83.33%,而采用实时荧光定量PCR检测方法,全部检出副溶血性弧菌。对分离到的50株副溶血性弧菌进行耐热溶血素(tdh)基因检测,结果4株tdh阳性。监测结果表明舟山贝类海产品中携带副溶血性弧菌情况比较严重,应引起足够的重视。     

Detection of Vibrio parahaemolyticus in Mediterranean mussels (Mytilus galloprovincialis) in Slovenia

The aim of this study was to determine the prevalence of Vibrio parahaemolyticus in shellfish samples harvested along the Slovenian coast. Shellfish samples of Mediterranean mussels (Mytilus galloprovincialis) were collected along the Slovenian coast at four locations (Se?a, Piran, Strunjan and Debeli Rti?) between 2006 and 2008. Samples were examined and analysed for the presence of V. parahaemolyticus by conventional and molecular methods. The presence of Vibrio in the samples was examined by conventional methods on plate grown bacterial cells before and after enrichment in alkaline saline peptone water (ASPW). PCR methods were used for the detection of V. parahaemolyticus-specific toxR and tlh genes and of the virulence-associated tdh and trh genes. Out of 168 samples examined, 24 were positive for toxR and tlh genes by PCR from enrichment broth. Five out of 62 (8.1%), 4 out of 32 (12.5%) and 15 out of 74 (20.2%) samples were positive in 2006, 2007 and 2008, respectively. Colonies of V. parahaemolyticus were isolated from only one sample positive for V. parahaemolyticus by PCR.     

Attachment of Vibrio parahaemolyticus strains to estuarine algae.

Attachment of Vibrio parahaemolyticus strains to estuarine microalgae was examined in artificial seawater by viable counts of the organism and direct counts of the bacterial cells after immunoperoxidase staining. Thermostable direct hemolysin (TDH)-producing and TDH-non-producing strains of V. parahaemolyticus were found to attach to five estuarine strains of Navicula (diatom alga) in similar levels. The level of the bacterial attachment depended on salinity and temperature of the water, in which the maximum attachment was observed in 15% artificial seawater at 25 degrees C, a typical condition of Hashizu estuary in Japan during summer months. The attachment was inhibited by pectinase digestion of the algal cells. These evidences confirmed the participation of the microalgae to the ecological cycle of V. parahaemolyticus at the estuary.     

荧光定量PCR法检测副溶血弧菌tdh基因的表达差异

以pvuA为内标基因,运用实时荧光定量PCR检测不同来源以及不同应激条件下副溶血弧菌热稳定直接溶血素基因tdh的表达量。pvuA和tdh基因的荧光定量PCR融解曲线分析表明,两者均为特异性扩增。尽管相同来源的不同菌株间tdh表达量存在显著差异,副溶血弧菌临床分离株的tdh mRNA平均表达量显著高于海产品分离株((57.2比13.8)。在pH4.0、0.5%和8%NaCl应激条件下,临床株ZJ2和海产品分离株FJ14A的tdh mRNA表达量显著高于对照组;另一海产品分离株KP34在8%NaCl条件下的表达量显著提高,而低pH应激时tdh mRNA的表达量显著降低。结果表明,不同副溶血弧菌分离株的tdh mRNA表达差异显著,临床分离株的tdh mRNA表达量总体上高于海产品分离株,副溶血弧菌在不同应激条件下主要表现为tdh mRNA表达上调。     

副溶血弧菌海产品分离株及临床分离株的多位点序列分型

选取毒力调控基因toxRS和看家基因gyrB、recA作为靶基因,对浙江沿海地区40株副溶血弧菌海产品分离株与8株临床分离株进行多位点序列分型。toxRS的多态性位点比例(10.2%)虽低于gyrB(12.0%)与recA(25.4%),但与gyrB均可分辨出最多的序列型(38),具有最强的分辨力(0.986)。3个基因串联后可分出44个序列型,分辨力达0.994。副溶血弧菌分离株呈现出较大的多样性。各地的海产品分离株分布于A群、B群,而临床分离株则主要集中于A群;C群仅包括1个临床分离株。其中临床分离株C2、C5、C7与海产品分离株F24属于同一个序列型,由此可推测该序列型在地域上分布较为广泛并可引起人发生胃肠炎等。因而副溶血弧菌所引起的对公共卫生的潜在风险性不容忽视。     

北海近岸养殖海域3种弧菌的分离鉴定及耐药性分析

对北海近岸养殖海域的3种弧菌(Vibrio)进行了分离鉴定,并对鉴定的弧菌进行耐药性分析。通过从北海近岸养殖海域随机采集水样,利用TCBS培养基对所采水样进行弧菌的分离和纯化,采用PCR方法和序列分析对弧菌进行鉴定,并对鉴定出的3种弧菌进行了常用抗菌药物耐药性的检测。共分离获得67株疑似弧菌菌株,经鉴定,19株为副溶血性弧菌(Vibrio parahaemolyticus),4株为霍乱弧菌(Vibrio cholera),4株为河流弧菌(Vibrio fluvialis)。耐药性分析研究表明,19株副溶血性弧菌总体显示对10种常用抗菌药物具有耐药性,其中100%的菌株对四环素、卡那霉素和红霉素表现耐药,94.7%的菌株对氨苄西林耐药、84.1%的菌株对左氧氟沙星和78.9%的菌株对环丙沙星耐药;4株霍乱弧菌和4株河流弧菌对10种抗菌药物的耐药性均较强,但敏感度低,两者对复方新诺明均表现为中介敏感。本研究提示,北海近岸养殖海域中弧菌种类较多,其中以副溶血性弧菌为主,大多对常用抗菌药物具有耐药性,对北海近岸海水养殖疾病防治具有一定的指导意义。     

Multiplex PCR and RPLA Identification of Staphylococcus aureus enterotoxigenic strains from bulk tank milk

Enterotoxigenic Staphylococcus aureus in raw milk poses a potential health hazard to consumers, and the identification of such strains should be used as part of a risk analysis of milk and milk products. The primary purpose of this study was to investigate the occurrence of enterotoxigenic S. aureus strains in raw milk supplied for dairy processing in the Czech Republic. A further aim was to compare the production of staphylococcal enterotoxins (SEs) with the presence of the corresponding genes. This was undertaken using multiplex polymerase chain reaction (PCR) and reversed passive latex agglutination (RPLA). Out of 440 bulk tank milk samples from 298 dairy herds, 70 proved positive for S. aureus (15.9%). Staphylococcal enterotoxin genes (ses) were detected in 39 (55.7%) isolates. The genes most commonly detected were sei (38.6%), seg (31.4%) and sea (27.1%). Genes seb, seh, sed, sej and sec were observed in 10%, 4.3%, 2.9%, 2.9% and 1.4% of strains respectively. Genes see and sel did not occur. The most frequently detected genotypes were seg, sei at 11.4%; sea at 10.0%; and sea, seg, sei at 8.6%. Toxin production was observed in nine (12.9%) S. aureus isolates. Seven strains were detected as SEB- (10%) and two as SED- (2.9%) producing. A relatively high number (32%) of discrepancies between the results with multiplex PCR and RPLA assays was obtained, particularly on account of SEA. Nineteen strains were sea positive by PCR but SEA negative by RPLA, and one strain was sec positive and SEC negative. The results of both methods were identical concerning SEB and SED. It was concluded that detection of ses by PCR was a useful additional tool to support identification of enterotoxigenic strains.