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.     

吉林省肉及肉制品食源性致病菌检测与分析

目的 了解吉林省肉及肉制品食源性致病菌污染情况,分析主要的危险因素,为防控食源性疾病提供科学依据。方法 按照随机采样原则和食源性致病菌监测样本采样要求,对2011-2019年吉林省9个市的肉及肉制品采集5683株样品,依据GB/T 4789的标准方法,对沙门氏菌、金黄色葡萄球菌、单核细胞增生李斯特氏菌进行检测。结果 共检出阳性食源性致病菌314株, 总体检出率为5.53%, 其中沙门氏菌、金黄色葡萄球菌、单核细胞增生李斯特氏菌检出率依次为1.47%、5.51%、9.41%。在不同地点采集的标本中,农贸市场、酒店和超市检出率较高,集体食堂和学校周围小商铺均未检出。结论 吉林省各市不同场所的肉及肉制品存在不同程度的食源性致病菌污染,食品安全监管部门应根据污染程度实施有针对性的食品安全监督管理。     

Alliinase-independent Inhibition of Staphylococcus aureus B33 by Heated Garlic

ABSTRACT: Heated (121 °C) garlic extract in which alliinase was inactivated before crushing exhibited complete bacteriostatic activity at 15% against Staphylococcus aureus. Garlic heated for 45 min showed the highest antibacterial activity and the relative peak areas of 4-heptenal, methyl allyl disulfide, diallyl disulfide, 2-vinyl-4H-1,3-dithiin, and diallyl trisulfide (DATS) were highest at 45 min of heating. Other than 4-heptenal, all these compounds are known to possess different degrees of antibacterial activity. DATS was thought to be the primary antibacterial compound in heated garlic extract. It was tentatively concluded that antibacterial compounds were formed from alliin unreacted by alliinase by marked heating. Diallyl compounds with more than three sulfur atoms and the oxidized form of allicin were not detected.     

合肥市即食肉类食品中金黄色葡萄球菌污染情况及耐药和基因型分布

目的调查合肥市即食肉类食品中金黄色葡萄球菌的污染情况,并对分离株进行耐药性分析及多位点序列分型研究。方法 2017年10月—2018年10月自合肥市区范围内的超市和开放型菜市场共采集744份即食型肉类食品样品。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》中的方法对样品进行金黄色葡萄球菌定性和定量分析,采用K-B纸片扩散法检测分离株的耐药性,采用多位点序列分型法对分离株进行ST分型,使用MEGA 7.0绘制聚类树状图。结果样品中金黄色葡萄球菌检出率为4.2%(31/744),其中卤肉制品检出率最高为8.1%(15/185),其次为烤肉制品(6.3%,11/174),定量分析显示超过90%的样品金黄色葡萄球菌污染水平在0.3～10 MPN/g范围内。耐药性分析结果显示80.6%(25/31)的菌株对青霉素G耐药,40%以上的菌株对链霉素、红霉素和卡那霉素耐药;多重耐药菌株占比为61.3%(19/31)。分型分析共获得8个ST型,分别为ST6、ST7、ST15、ST59、ST72、ST188、ST398和ST2196型,其中以ST188和ST72型为主。结论合肥市卤肉和烤肉制品中存在金黄色葡萄球菌污染,且多重耐药菌株比例较高,对消费者的健康有潜在威胁,有必要持续监测污染率、菌株耐药和基因型流行趋势,完善合肥市食源性金黄色葡萄球菌感染的风险评估。     

食源性微生物MRSA及其检测方法在食品安全中的研究进展

食品安全在世界卫生组织的定义是指所有食品中有毒、有害物质对人体健康影响的公共卫生问题。2008年"三聚氰胺"事件后,在近几年的全国两会上,食品安全问题连续成为焦点话题;而同期全国人大常委会通过的《食品安全法》,更显示了食品安全的严重与令人担忧,其监测与控制显得迫在眉睫。作为典型的食源性微生物,金葡菌尤其是耐甲氧西林金黄色葡萄球菌(Methicillin-resistant Staphylococcus aureus,MRSA)其新型的耐药特点显示了细菌耐药性的发展和进化可能是由于大量抗生素在畜牧业中的滥用而造成的。由于抗菌药物的研制周期跟不上耐药性出现的速度,使得细菌治疗面临越来越严峻的考验,甚至出现无药可用的情况,严重威胁人类健康。本文结合食品安全抗生素滥用问题与最近出现的超级细菌耐药问题,根据在相关领域多年的研究数据,对常见的食源性微生物MRSA及其检测方法在食品安全中的研究进展进行综述。     

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.     

紫甘薯花色苷色素抑制金黄色葡萄球菌作用初探

本实验研究紫甘薯花色苷色素对金黄色葡萄球菌的抑制作用.结果表明,紫甘薯花色苷色素对金黄色葡萄球菌抑制的最低浓度为200μg/ml.金黄色葡萄球菌生长曲线和透射电镜观察表明,紫甘薯花色苷的抑菌作用可能是通过增强细胞膜的通透性,使细胞异常生长,抑制对数生长期的细胞分裂,使细胞质稀薄、解体,导致细胞死亡.SDS-PAGE分析表明,紫甘薯花色苷对大肠杆菌和金黄色葡萄球菌蛋白表达影响不明显,未见特征性条带的消失,仅对部分蛋白质合成量有影响.     

Activity of Silver Nanoparticles against Staphylococcus spp.

Staphylococcus epidermidis is a bacterium that is part of the human microbiota. It is most abundant on the skin, in the respiratory system and in the human digestive tract. Also, Staphylococcus aureus contributes to human infections and has a high mortality rate. Both of these bacterial species produce biofilm, a pathogenic factor increasing their resistance to antibiotics. For this reason, we are looking for new substances that can neutralize bacterial cells. One of the best-known substances with such effects are silver nanoparticles. They exhibited antibacterial and antibiofilm formation activity that depended on their size, shape and the concentration used. In this review, we presented the data related to the use of silver nanoparticles in counteracting bacterial growth and biofilm formation published in scientific papers between 2017 and 2021. Based on the review of experimental results, the properties of nanoparticles prompt the expansion of research on their activity.