金黄色葡萄球菌食品分离株肠毒素基因分布及分型研究

目的 对2006-2009年上海市Staphylococcus aureus(S.aureus)食品分离株进行肠毒素基因检测和基因分型研究,以了解肠毒素基因的分布规律及S.aureus的流行特点.方法 利用PCR方法检测食品中金黄色葡萄球菌肠毒素基因,包括5种传统肠毒素基因(SEA-SEE)和4种新型肠毒素基因(SEG-SEJ);利用脉冲场凝胶电泳法对49株食品分离株进行基因分型.结果 本研究发现49株食品分离株中有19株含有肠毒素基因,16株含有传统肠毒素SEA和SEC,且SEC占93.8％,并检测到新型肠毒素SEG、SEI、SEJ和SEH.PFGE法基因分型显示5株菌不能被分型,其余44株可分为28个基因型,表现为基因型的多样性,且分离自不同时间的菌株具有相同的带型.结论 应加强食品中S.aureus的监测分析,为其引起的食物中毒的预防和控制提供科学依据.     

鄂西北地区食源性金黄色葡萄球菌污染及耐药性和毒力基因分析

目的 了解鄂西北地区食源性金黄色葡萄球菌的检出情况、耐药性以及毒力基因分布特征。方法 从湖北省襄阳市、十堰市、随州市三地共采集303份食物样品进行金黄色葡萄球菌筛查,通过聚合酶链式反应(PCR)法进行耐甲氧西林金黄色葡萄球菌(methicillin-resistant Staphylococcus aureus,MRSA)的鉴定和毒力基因检测,并用K-B纸片扩散法进行耐药性测定。结果 303份样品中共有41份检出金黄色葡萄球菌,阳性率为13.53%,其中生肉制品检出率最高(23.91%,22/92)。在产肠毒素菌株中以携带产肠毒素基因sea的菌株最多,占87.80%(36/41)。携带表皮剥脱素基因eta和中毒性休克综合征毒素基因tst的菌株分别占97.56%(40/41)和7.32%(3/41)。同时携带3种及以上肠毒素基因的菌株占17.07%(7/41)。同时含有eta和tst的菌株占2.44%(1/41)。药敏结果显示,分离菌株对青霉素、四环素、红霉素和多西环素的耐药率分别为78.05%(32/41)、43.90%(18/41)、31.71%(13/41)和21.95%(9/41),对其他8种抗菌药物的耐药率均<20%。mecA基因检测表明,19.51%(8/41)的菌株为MRSA,且7株MRSA菌株来源于生肉制品和未加调料的素卤菜。结论 鄂西北地区食源性金黄色葡萄球菌具有较高的检出率,而且产毒株较多,并且存在多重耐药现象,相关部门需加强食品安全监测以控制该菌株的流行与扩散。     

Nitric Oxide Complex of Iron(II) Myoglobin Converted from Metmyoglobin by Staphylococcus xylosus

With Staphylococcus xylosus FAX-1, metmyoglobin in MRS broth (pH 5.8) was found to undergo conversion to hexacoordinate nitric oxide (NO) complex of Fe(II) myoglobin. When the pH of the MRS culture containing myoglobin changed from 5.8 to 4.0, it affected the conversion from hexacoordinate to pentacoordinate NO complex of Fe(II) myoglobin. This conversion process was reversible. Salami without nitrite or nitrate addition was prepared by inoculating S. xylosus FAX-1, and pentacoordinate NO complex of Fe(II) myoglobin (nitrosylmyoglobin formed in cured meat) was formed in the salami.     

广州市白云口岸航空食品中金黄色葡萄球菌凝固酶基因分型及肠毒素基因研究

目的对2013—2015年从广州市白云口岸航空食品中分离的金黄色葡萄球菌进行基因分型研究,为食源性金黄色葡萄球菌分子溯源提供基础数据。方法以血浆凝固酶和肠毒素为目标基因,采用聚合酶链式反应(PCR)方法对9株金黄色葡萄球菌进行基因分型,其中6株为航空食品分离株,1株为配餐车间大门手拭分离株,2株为标准菌株。肠毒素基因检测包括5种传统肠毒素基因(sea、seb、sec、sed、see)和6种新型肠毒素基因(ser、seg、seh、sei、sej、sep)。结果 6株航空食品分离株的血浆凝固酶基因扩增分型结果为2个PCR型,酶切后得3种亚型;肠毒素基因检测结果显示有2株航空食品分离株含有肠毒素基因,检出率为33.3%(2/6),检出的基因为2种传统肠毒素基因(sec、sed)和4种新型肠毒素基因(ser、seg、sei、sej),均同时携带2种以上肠毒素基因。结论血浆凝固酶基因扩增分型结果显示,不同时间、不同采集地点存在相同的基因型,提示金黄色葡萄球菌存在交叉污染的可能性;航空食品分离株共检出6种肠毒素基因,提示金黄色葡萄球菌基因型多样性,应加强其他新型肠毒素基因检测。     

葡萄球菌和乳酸菌对广式腊肠风味的影响

本文研究了葡萄球菌、乳酸杆菌单独发酵和混合发酵对广式腊肠生产过程中风味变化的影响,并考察了不同发酵时间对成品腊肠风味及相关理化指标的影响,旨在研究以微生物发酵技术提高广式腊肠的风味。结果显示,葡萄球菌(M1、M14)和乳酸杆菌(PED、STR)单独或混合发酵均能在一定程度上提高广式腊肠的腊香味,混合菌种比单一菌种发酵效果好。随着生产时间的推移腊肠香味逐渐显现,烘烤至成品后腊肠风味的感官分值最高。腊肠生产过程的pH值随菌种或配比的不同而有不同的变化,pH过低对腊肠风味有不利的影响。添加菌种处理组在不同的发酵天数所对应的广式腊肠成品的挥发性盐基氮、氨基态氮、酸价都比空白对照组的高,而过氧化值比空白对照组的低,证明所选用菌种的发酵对腊肠的蛋白质和脂肪的水解有一定的促进作用。研究结果表明当菌种的混合配比为葡萄球菌(M1∶M14=1∶1):乳酸杆菌(PED:STR=1∶1)=1∶100时,发酵2d后广式腊肠的风味最好。     

Phenotypic and Genotypic Antimicrobial Resistance Traits of Foodborne Staphylococcus aureus Isolates from Shanghai

Staphylococcus aureus is a recognized pathogen in humans, which causes nosocomial infections and food poisoning. The transmission of antibiotic resistant S. aureus (ARSA), especially methicillin‐resistant S. aureus, between food products and humans has become a serious problem. Hence, it is necessary to monitor S. aureus through the food supply chain. In this study, the disk diffusion method and epsilometer test were performed to determine the prevalence of ARSA in 78 foodborne isolates using 18 antibiotics. The highest resistance frequency was found for penicillin G (74.4%), followed by erythromycin (59.0%) and clindamycin (44.9%), whereas no vancomycin‐resistant isolates were found. The 78 isolates could be subtyped into 31 resistance profiles and 11 clusters based on their antimicrobial susceptibility. Furthermore, Polymerase chain reaction (PCR) screening for the presence of 13 genes conferring antibiotic resistance was conducted. The presence of resistance genes was relatively high: bla_TEM (80.8%), ermB (41.0%), grlA (38.5%), ermC (35.9%), and aac6’/aph2” (35.9%). The incidence of antibiotic resistance was significantly correlated to food types (p = 0.018), with isolates from meat and raw milk more resistant to antibiotics than those from frozen food and vegetables.     

多重实时荧光定量PCR快速检测婴幼儿奶粉中沙门氏菌、克罗诺杆菌和金黄色葡萄球菌

目的建立多重实时荧光定量PCR(multiplex quantitative real-time PCR,multiplex qPCR)快速检测奶粉中金黄色葡萄球菌、沙门氏菌和克罗诺杆菌3种常见致病菌的方法。方法筛选目标菌株的特异性引物与探针,优化反应体系,建立稳定的多重q PCR反应体系。通过阳性菌株加标的方式验证体系的特异性,并确定人工污染奶粉的检出限。结果各对引物探针对目标菌株均能扩增,多重实时荧光PCR未发现交叉反应,对17株非目标菌进行检测均未检出,人工污染奶粉中克罗诺杆菌和沙门氏菌的检出限均为10~3 CFU/mL,金黄色葡萄球菌的检出限为10~4 CFU/mL。结论本研究方法可实现婴幼儿奶粉样品中3种致病菌qPCR高效率检测。     

Systematic survey on the prevalence of genes coding for staphylococcal enterotoxins SElM, SElO, and SElN

Staphylococcus aureus remains a leading cause of food-poisoning with substantial impact on public health. Using a multiplex polymerase chain reaction-DNA enzyme immunoassay (PCR-DEIA), we studied the presence of genes encoding staphylococcal enterotoxin-like (SEl) superantigens sem, sen, and seo, associated with the enterotoxin gene cluster (egc), in 429 clinical Staphylococcus aureus isolates. 294 (68.5%) isolates tested positive for at least one of the three SEl genes. In contrast to the fixed gene combination seg/sei also located on egc, a substantial number of isolates (n = 108) were found to bear only one or two of the genes encoding SElM, SElN, and SElO. Regarding the origin of the S. aureus isolates, a significant difference (P = 0.022) was found for the possession of seo (61.2% of blood isolates versus 42.9% of nasal strains). Also sem (not significantly) was found more common in blood isolates (52.1% versus 40.5%). The survey of the newly described SEl genes sem-seo supports the concept that most clinical S. aureus isolates harbor subsets of pyrogenic toxin superantigens. The potential contribution of seo and sem to the pathogenic potential of S. aureus has to be further evaluated.     

四川香肠中产香葡萄球菌的分离与鉴定

从四川自然发酵香肠中共分离得到242株微球菌和葡萄球菌,采用快速半定量平板方法从中筛选出了硝酸盐还原酶活性较大的株菌80株,并对这些菌株进行发酵基本实验和产香基本要求实验,筛选得到了2株过氧化氢酶阳性葡萄球菌S15和S25。通过GC-MS分析发现,在实验条件下只有S25具有代谢亮氨酸生成3-甲基丁醛的能力。生化鉴定以及菌株的16S rDNA测序结果表明,S25为腐生葡萄球菌。     

2016~2019年吉林省食源性金黄色葡萄球菌监测数据分析

目的 探究2016~2019年吉林省食源性金黄色葡菌球菌污染现状。方法 采集2016~2019年吉林省9个地(市)级的餐厅、超市和零售市场等地点食品样本,共计2824件,根据GB 4789.10-2016《食品安全国家标准 食品微生物学检验 金黄色葡萄球菌检验》进行测定,采用率、构成比和X2检验的方法对检出率进行统计描述和分析。结果 2016~2019共监测样本2824份,总检出率2.02%;9个地区中白城市总检出率最高（4.59%）,其次为四平市（3.82%）,白山市总检出率最低（0.51%）;13类食品中有8类食品检出金黄色葡萄球菌,分别为焙烤及油炸类食品、餐饮食品、调味品、豆制品、冷冻饮品、肉及肉制品、乳与乳制品和速冻米面食品,其中速冻米面食品中金黄色葡萄球菌检出率最高（9.00%）;餐饮服务环节阳性检出率为2.45%（22/898）,流通环节阳性检出率为1.87%（36/1928）;CFU(colony forming units,平板计数法)法测得金黄色葡萄球菌检出结果的中位数[四分位数间距]为5.29[2.65~7.94] CFU/g(mL),MPN(Most Probable Number,最大可能数法)法测得金黄色葡萄球菌检出结果的中位数[四分位数间距]为1.53[0.76~5.29]MPN/g(mL)。结论 2016~2019年吉林省各地市食品中金黄色葡萄球菌污染情况普遍存在,其中白城市污染相对严重,吉林省所有食品类别中,速冻米面制品污染相对严重,应给予高度重视,餐饮服务环节和流通环节还需加强监管。