应用LAMP方法检测动物源性食品中沙门菌

目的运用两种方法对动物源性食品中沙门菌进行检测,找出一种更加快速、敏感、特异、简便的检验方法。方法采集60份市售的禽、畜等生肉及60份奶制品,分别采用LAMP方法和国家标准方法GB/T4789.4—2008对沙门菌进行分离与鉴定。结果 120份采集样品中,LAMP方法检出2例,阳性率1.67%;国家标准方法检出1例,阳性率0.83%。LAMP方法的符合率为99.2%,敏感性为100%,特异性为99.2%。结论 LAMP检测法快速、特异、简便,该方法与国标法的阳性率差异无显著性。     

2010年江苏省肉鸡沙门菌污染专项监测分析

目的了解肉鸡养殖和屠宰过程中沙门菌的污染状况,为确定从生产到销售各环节沙门菌分布和疾病可能传染源、制定公共卫生措施并评价其有效性提供科学依据。方法按照《全国食源性致病菌监测工作手册》中专项监测的采样和实验室检测要求,2010年共采集活体肉鸡肛拭样本210份,胴体样本204份,监测沙门菌污染情况。结果肛拭样本检出沙门菌23份,检出率10.95%;胴体样本检出沙门菌71份,检出率34.80%。两类样本的血清学分型构成比不同(χ2=15.7,P<0.001),肛拭样本检出的23株沙门菌中有22株印第安纳沙门菌,胴体样本中检出的71株沙门菌中有35株为印第安纳沙门菌,22株为奥尔巴尼沙门菌。结论肉鸡活体肛拭和胴体样本沙门菌检出率差异有统计学意义(χ2=33.5,P<0.001),胴体高于活体检出率,活体和胴体的沙门菌来源不同,胴体可能存在交叉污染。     

肉猪养殖和屠宰环节沙门菌污染状况监测分析

目的 了解开封市肉猪养殖屠宰加工过程中沙门菌的污染状况.方法 采集不同来源的养殖场猪混合粪便、成年猪肛拭子、胴体涂抹物、肠系膜淋巴结样本,共187份,按照GB 4789.4-2010进行沙门菌的分离和鉴定.结果 猪混合粪便沙门菌检出率为4.5％,肛拭子沙门菌的检出率为16.4％,胴体涂抹物沙门菌检出率为34.5％,肠系膜淋巴结沙门菌检出率为27.3％.不同季节的检出率差异有统计学意义(P＜0.05).阳性分离株分布于沙门菌的5个群,共14个血清型,以阿贡纳沙门菌和德尔卑沙门菌为主.结论 屠宰环节是沙门菌交叉污染的关键点,应采取综合措施,有效控制猪肉沙门菌污染.     

两种选择性培养基HE、CAS检测沙门菌效果的比较

目的 比较两种沙门菌选择性培养基HE和CAS在食物样本检测中的敏感性、准确性和时效性.方法 观察典型沙门菌和非沙门菌在两种选择培养基上的生长状况和菌落形态.通过人工染菌实验,比较两种选择性培养基分离沙门菌的检测效果.采集16份市售整鸡样本,进一步比较两种培养基沙门菌的阳性检出率.结果 沙门菌在HE平板上为绿色菌落,大多有黑心;在CAS平板上为紫色菌落.含菌量为10 CFU/25 g鸡肉样本,经SC增菌液增菌培养8h,可用CAS检出沙门菌;而采用HE则需增菌18 h才可检出.16份整鸡样本,增菌8,12和18 h,采用CAS的阳性检出率分别为43.75%、31.25%和37.5%,相应时间下采用HE的阳性检出率分别为18.75%,25%和18.75%,8份新鲜整鸡样本,检出4份阳性;8份冷冻整鸡样本,检出3份阳性.结论 CAS选择性平板可抑制大多杂菌生长,沙门菌在其上菌落形态较易鉴别.CAS分离沙门菌的检出时限和检测灵敏度都明显优于HE,建议在食物样本检测沙门菌时应首选CAS.     

济南市零售鸡肉中沙门菌污染状况及可移动黏菌素耐药基因分析

目的 了解2020—2021年济南市零售鸡肉中沙门菌的污染状况,并探究可移动黏菌素耐药基因（mcr）的携带情况。方法 2020年12月至2021年11月在济南市采集零售鸡肉样品260份,依据GB 4789.4—2016《食品安全国家标准食品微生物学检验沙门氏菌检验》对样品中的沙门菌进行分离鉴定,通过聚合酶链式反应（PCR）对分离株进行血清型鉴定和mcr基因筛选,使用微量肉汤法对mcr基因阳性株开展抗生素敏感性试验。结果 260份零售鸡肉样品中共检出阳性样品61份,污染率为23.46%(61/260),秋季污染率最高可达53.33%(32/60);共分离出沙门菌103株,56株为肠炎沙门菌,占比54.37%(56/103)。2株不同产地鸡翅样品来源的印第安纳沙门菌分离株检测出mcr-1基因,阳性率为1.94%(2/103)。2株mcr-1基因阳性印第安纳沙门菌分离株均为多重耐药株,其中1株可对碳青霉烯类和多黏菌素类在内的全部12类测试抗生素同时耐药。结论 2020—2021年济南市零售鸡肉中的沙门菌污染较为严重,秋季采集样品的污染率较高,肠炎沙门菌为优势血清型,检出携带mcr-1基因;同...     

应用SE-μPADs法检测库尔勒市动物源食品中沙门菌的效果评价

目的:为了快速检测动物性食品中沙门菌的污染。方法:应用国家标准方法和SE-μPADs快速检测方法,对库尔勒市场及屠宰场中的380份样品进行沙门菌检测的评价。结果:国家标准方法中选择性培养基的初筛率为20.00%(76/380),从初筛菌株中分离鉴定出6株沙门菌;SE-μPADs快速检测方法的初筛率为6.58%(25/380),最终分离鉴定出10株沙门菌。SE-μPADs快速检测方法与国家标准方法阳性符合率为100%。库尔勒市场及屠宰场中共检测出10株沙门菌,总检出率为2.63%,而肉样污染率最高,胴体拭子次之,污染率分别达3.66%、3.31%。结论:SE-μPADs快速检测方法的初步应用效果较好,适合推广应用,且库尔勒市应加强对屠宰场及市场上肉品中沙门菌的污染监测。     

新疆零售牛肉中沙门菌毒力基因的检测

目的了解新疆库尔勒市零售牛肉沙门菌污染状况和沙门菌分离株毒力基因携带状况。方法参考食品安全国家标准GB 4789.4-2016的方法,采用培养基培养和沙门菌生化鉴定试剂盒对零售牛肉中的沙门菌进行分离鉴定,并对沙门菌分离株10种毒力基因(inv A、hil A、ssa Q、mgt C、sii D、sop B、spv B、spv C、spv D、spv R)进行PCR检测。结果 317份牛肉样品共检出沙门菌阳性样品28份,分离鉴定到阳性沙门菌28株,污染率为8.8%(28/317);除ssa Q、spv C毒力基因外,其余8种毒力基因均有检出。毒力基因inv A和spv R具有高度的稳定性,携带率均为100%(28/28);毒力基因sii D、spv B和spv D的携带率也较高,分别达78.6%(22/28)、75.0%(21/28)和75.0%(21/28)。结论新疆库尔勒市零售牛肉沙门菌污染较为严重,应采取措施加强对新疆库尔勒市牛肉生产各环节的防控与食品卫生监督。     

2006年东阳市食品致病菌污染状况监测分析

目的对东阳市食品中6种致病菌的分布状况进行监测,确定污染致病菌的高危食品,为食物中毒监测提供科学依据。方法依据国标方法和2006年中国疾病预防控制中心食品污染菌调查工作手册,采用进口显色培养基、生物梅里埃-ATB鉴定系统,对采集的食品样本分别进行沙门菌、单核细胞增生性李斯特菌、EHEC O157：H7、弯曲菌、金黄色葡萄球菌和副溶血性弧菌的分离和生化鉴定。结果检测生肉（牛、羊、鸡、猪）、熟肉、非生食水产品、生食水产品、生食蔬菜、蔬菜色拉、鲜榨果汁共7类样品143份,检出致病菌38株（其中有2份生猪肉同时检出2株沙门菌）,总检出率25.2％。其中分离到副溶血性弧菌6株,金黄色葡萄球菌12株,沙门菌17株,单核细胞增生性李斯特菌3株。EHEC O157：H7和弯曲菌未栓出。7类食品中致病菌阳性率最高的为非生食水产品（70．0％）,其次为蔬菜色拉（50.0%）、生肉（39.5％）、熟肉制品（15．2％）,生食蔬菜和生食水产品中均未检出致病菌。结论东阳市居民主要消费食品存在食源性致病菌污染,其中非生食水产品和生肉是主要被污染食品。主要污染致病菌是副溶血性孤菌、金黄色葡萄球菌和沙门菌。     

扬州市市售鸡肉中弯曲菌定性定量流行病学分析

目的调查分析超市和农贸市场零售鸡肉中弯曲菌定量污染水平,为弯曲菌污染的定量风险评估、危害分析和防控提供数据。方法选择扬州市内各大型超市和农贸市场作为采样点,采取未分割的鸡胴体,在3 h内送到实验室开展检测。运用选择性CCDA平板直接计数法对弯曲菌进行定量检测,涂布平板法计数,用PCR进行鉴定。结果采集的40只整鸡,检测出阳性样品19份,平均阳性率为47.5%,平均带菌量为229.2 CFU/g。其中超市的8份阳性整鸡中,平均带菌量为28.3 CFU/g;农贸市场的11份阳性整鸡中,平均带菌量为313.6CFU/g。共分离出56株弯曲菌,其中空肠弯曲菌29株,结肠弯曲菌23株,其他弯曲菌4株。结论定性及定量结果显示,零售鸡中存在弯曲菌污染,农贸市场的零售鸡肉比超市的零售鸡肉污染严重,需要加强防控措施。     

六类食品中致病性细菌检测

为了解昌平区细菌对食品污染的特点，2004年采集6类305件食品，按照GB／T4789－2003食品微生物检验标准进行沙门菌、金黄色葡萄球菌、副溶血性弧菌、单核细胞增生性李斯特菌、0157：H7大肠杆菌5种致病菌检测。菌株鉴定应用API细菌鉴定系统，肠毒素检测用minVIDAS测试系统，单核细胞增生性李斯特菌毒力基因测定采用聚合酶链反应（PCR）。金黄色葡萄球菌检出率为16．07％，其中生肉、生牛奶、生食水产品检出率比较高，分别为25％、16．98％、27．27％；散装熟肉制品、乳制品也有检出，分别为4.29％、5％。在检出的49株金黄色葡萄球菌菌株中有31株产生肠毒素，占63．26％，其中生、熟肉类肠毒素阳性25株；从奶类、生食水产品中各检出3株。单核细胞增生性李斯特菌检出34株，检出率为11．15％，其中生肉检出率高达20％，其次为散装熟肉制品（8．57％）；另外从淡水鱼中检出4株单核细胞增生性李斯特菌。28株单核细胞增生性李斯特菌毒力基因（hly、iap、prfA）检测全部阳性。生肉中检出一株肠炎沙门菌。未检出O157：H7大肠杆菌、副溶血性弧菌。昌平地区金黄色葡萄球菌和单核细胞增生李斯特菌对食物的污染比较严重，生食品与加工后食品的检出率差异有显著性。     

Prevalence and antimicrobial susceptibility of Salmonella isolated from a variety of raw meat sausages in Gaborone (Botswana) retail stores

The objective of the study was to provide baseline data on the prevalence and antimicrobial susceptibility of Salmonella in different types of raw meat sausages directly accessible to the consumers in Gaborone, Botswana. A total of 300 raw sausages comprising 79 beef, 78 pork, 72 chicken, and 71 mutton samples were concurrently analyzed for the presence of Salmonella using a conventional culture method and a validated PCR method. The PCR assay results were in full concordance with those of the conventional culture method for the detection of Salmonella. Sixty-five (21.7%) of 300 samples were positive for Salmonella by both the conventional culture method and PCR assay. Even though more chicken samples contained Salmonella than did any other sausage type, the difference in the presence of Salmonella among the four sausages types was not significant. Eleven serotypes were identified, and Salmonella enterica subsp. salamae II was most prevalent in all the sausage types. Beef sausages generally had higher mesophilic bacterial counts than did the other three sausage types. However, higher microbial counts were not reflective of the presence of salmonellae. Susceptibility of the Salmonella enterica serotypes to 20 antimicrobial agents was determined, and Salmonella Muenchen was resistant to the widest array of agents and was mostly isolated from chicken sausages. Regardless of the meat of origin, all 65 Salmonella isolates were resistant to at least four antimicrobial agents: amikacin, gentamicin, cefuroxime, and tombramycin. This resistance profile group was the most common in all four sausage types, comprising 90% of all Salmonella isolates from beef, 71% from pork, 63% from mutton, and 35% from chicken. These results suggest that raw sausages pose a risk of transmitting multidrug-resistant Salmonella isolates to consumers.     

Campylobacter and Salmonella in raw red meats in the United Kingdom: prevalence, characterization and antimicrobial resistance pattern, 2003-2005

The prevalence of Campylobacter and Salmonella was assessed in 3959 raw red meats in the UK during 2003-2005. Meats were more frequently contaminated with Campylobacter (7.2%) than with Salmonella (2.4%). Lamb and other meats (e.g. mutton, rabbit) exhibited the highest contamination from Campylobacter (12.6% and 19.8%, respectively), compared with pork (6.3%) and beef (4.9%). Pork however had the highest contamination from Salmonella (3.9%), followed by lamb (2.0%), other meats (2.0%) and beef (1.3%). Offal samples (36.6%) were more frequently contaminated with Campylobacter or Salmonella than muscle tissue (7.0%). C. jejuni predominated in all meat types. C. coli isolates were more likely to exhibit antimicrobial drug resistance, including quinolones, than C. jejuni. Salmonella typhimurium was the most frequent Salmonella serotype isolated from meats; S. typhimurium DT104/104b isolates exhibited higher rates of multiple drug resistance than other serotypes. The findings reinforce the importance of adequate cooking of meat and good hygiene to avoid cross-contamination.     

Detection of Pork in Processed Meat by an Enzyme-linked Immunosorbent Assay Using Antiswine Antisera

An antiserum to autoclaved porcine muscle extract was raised in sheep and tested for the detection of pork in mixtures of pork in mutton (P/ S) or beef (P/B) heated at 70, 100, and 120°C for 30 min using a competitive enzyme-linked immunosorbent assay (ELISA) method. The results indicated that the antiserum detected low percentages of pork (1%) mixed in beef and in mutton (0.5%) even when the meat mixtures were heated at 70, 100 or 120°C for 30 min, which correspond to heat treatments of commercially processed meat products. Regression analysis showed a high positive correlation (r > 0.99) between absorbance (OD) values and different percentages of experimental or commercially processed P/B meat mixtures. It was concluded that the combination of anti-porcine sera raised in sheep and the ELISA method can be used for the detection of low percentages of pork in processed meats.     

Predicting pathogen growth during short-term temperature abuse of raw pork, beef, and poultry products: use of an isothermal-based predictive tool

A computer-based tool (available at: www.wisc.edu/foodsafety/meatresearch) was developed for predicting pathogen growth in raw pork, beef, and poultry meat. The tool, THERM (temperature history evaluation for raw meats), predicts the growth of pathogens in pork and beef (Escherichia coli O157:H7, Salmonella serovars, and Staphylococcus aureus) and on poultry (Salmonella serovars and S. aureus) during short-term temperature abuse. The model was developed as follows: 25-g samples of raw ground pork, beef, and turkey were inoculated with a five-strain cocktail of the target pathogen(s) and held at isothermal temperatures from 10 to 43.3 degrees C. Log CFU per sample data were obtained for each pathogen and used to determine lag-phase duration (LPD) and growth rate (GR) by DMFit software. The LPD and GR were used to develop the THERM predictive tool, into which chronological time and temperature data for raw meat processing and storage are entered. The THERM tool then predicts a delta log CFU value for the desired pathogen-product combination. The accuracy of THERM was tested in 20 different inoculation experiments that involved multiple products (coarse-ground beef, skinless chicken breast meat, turkey scapula meat, and ground turkey) and temperature-abuse scenarios. With the time-temperature data from each experiment, THERM accurately predicted the pathogen growth and no growth (with growth defined as delta log CFU > 0.3) in 67, 85, and 95% of the experiments with E. coli 0157:H7, Salmonella serovars, and S. aureus, respectively, and yielded fail-safe predictions in the remaining experiments. We conclude that THERM is a useful tool for qualitatively predicting pathogen behavior (growth and no growth) in raw meats. Potential applications include evaluating process deviations and critical limits under the HACCP (hazard analysis critical control point) system.     

Application of a multiplex PCR for the simultaneous detection of Escherichia coli O157:H7, Salmonella and Shigella in raw and ready-to-eat meat products

Escherichia coli O157:H7, Salmonella and Shigella might contaminate similar types of meat products and cause deadly diseases in humans. Traditional microbiological analyses to detect these pathogens are labor-intensive and time-consuming. The objective of this study was to apply a multiplex PCR for simultaneous detection of the pathogenic bacteria in certain raw and ready-to-eat meat matrices. The tested samples had aerobic plate counts ranging from non-detectable, in chicken nuggets and salami, to 8.36log(10)CFU/g in ground pork. The pH of homogenates spanned from 6.86, in ground beef, to 7.17 in salami. Following a 24-h enrichment, the multiplex PCR assay could concurrently detect the three pathogens at 0.2log(10)CFU/g in ground beef, roast beef, beef frankfurters, chicken nuggets, salami and turkey ham, and 1.2log(10)CFU/g in ground pork. This multiplex PCR offers an efficient microbiological tool for presumptive detection of E. coli O157:H7, Salmonella and Shigella in meat.     

PCR方法快速鉴别食品中肉的种类

本文针对食品中肉成分种类鉴别开发了一种快速灵敏的PCR检测方法,可检测食品中是否存在猪肉、牛肉、羊肉以及鸡肉等成分。采用微波助提法提取样品中DNA,简化了前处理步骤,可在短时间内完成从多种不同类型肉与肉制品中提取肉成分DNA。为了评价方法的可靠性与灵敏度,猪肉以及掺入了不同比例浓度猪肉成分的食品样品采用本方法进行了核酸提取与PCR分析。检测结果表明,方法可检测出低至含有0.5%浓度的猪肉成分的混合样品。随机抽取50份不同类型的市面食品样品,检测出5份食品含有猪肉成分,7份食品中含有牛肉成分,5份食品中含有羊肉成分。该样品前处理方法、DNA提取方法以及PCR检测方法可广泛应用于食品中肉成分种类的检测鉴别。     

Utilization of chickpea flour in sausages

'English' type fresh skinless sausages were prepared in which some of the meat (mutton, pork or beef) was replaced on a protein to protein basis by chickpea flour. The acceptability of mutton sausages containing chickpea flour was not affected at levels of substitution up to 40%, whereas pork and beef sausages were significantly less acceptable at substitution levels above 30%. In all the sausages incorporation of chickpea flour led to increased cooking losses and softer textures. Incorporation of chickpea flour caused discoloration of the raw sausages which became more prominent during storage at 0°C.     

Contamination of Salmonella in retail meats and shrimps in the Mekong Delta, Vietnam

From March 2000 to September 2001, 608 samples of retail meat (136 pork, 70 beef, 202 chicken, and 200 ducks) and 110 samples of retail shrimp from six provinces of the Mekong Delta in Vietnam were collected individually and examined for the prevalence of Salmonella. Of the 718 samples examined, 243 (33.8%) were Salmonella positive. Salmonella was isolated from 69.9% of the pork samples, 48.6% of the beef samples, 21.0% of the chicken meat samples, 22.3% of the duck meat samples, and 24.5% of the shrimp samples. From 261 Salmonella isolates, 24 different serovars were identified. The predominant serovars of the isolates were Salmonella Derby, Salmonella Weltevreden, and Salmonella London in pork; Salmonella Weltevreden, Salmonella London, and Salmonella Dessau in beef; Salmonella Emek, Salmonella Typhimurium, and Salmonella Dessau in chicken meat; Salmonella Lexington, Salmonella Derby, and Salmonella Dessau in duck meat; and Salmonella Weltevreden, Salmonella Tennessee, and Salmonella Dessau in shrimps. Salmonella Bovismorbificans, Salmonella Derby, Salmonella Dessau, and Salmonella Weltevreden were the most common serovars in all the samples examined. These results indicate a high rate of contamination by Salmonella in retail meats and shrimps in the Mekong Delta, Vietnam.     

2013年苏州地区肉及其制品掺假情况调查

了解苏州地区肉及其制品的掺假情况,通过对肉类种源与标签明示肉源进行比对,鉴别摻假食品,为加强食品标签管理提供依据。方法 运用自建的动物源性食品种源判定Taqman实时荧光PCR检测体系对苏州地区的肉及其制品进行种源判定,与标签明示肉源进行比对,鉴别摻假食品。结果 本次调查共检验涉及32个生产单位的90份样品,总不符合率为25.6%(23/90)。检测的44份牛肉及其制品中有12份与标签不符,8份用猪肉部分替代牛肉,1份以鸭肉部分代替牛肉进行销售;此外有3份不含有牛肉成分,存在猪、鸡、鸭源性肉类之外的肉类成分。共检测羊肉及其制品16份,有2份用鸭肉代替羊肉出售,3份羊肉样品中掺入了部分猪成分,其中1份样品还存在单个样品掺杂两种外源肉类的现象(猪源性和鸭源性)。检测猪肉及其制品19份,其中2份样品含有标签未注明的鸡肉成分。在所检测的11份混合肉类样品中有4份成分与标签不符,主要是以廉价的鸡肉取代/部分取代相对高价的牛肉和猪肉。结论 肉制品掺假情况明显,用猪肉、鸭肉部分代替牛肉和羊肉仍是主要的掺假手段,牛肉掺假样品主要是熟制牛肉制品,而火锅食用羊肉卷样品则是羊肉掺假高危品,开展肉制品摻假检测对规范肉制品市场具有积极意义。此外,3份未知种源成分的牛肉样品提示在现有检测基础上还需扩大检测范围,防患于未然。     

Belgian surveillance plans to assess changes in Salmonella prevalence in meat at different production stages

From 1997 to 1999, the prevalence of Salmonella was assessed at different stages through the pork, poultry, and beef meat production chains. Different dilutions of the initial sample suspension were analyzed to provide a semiquantitative evaluation of Salmonella contamination and to determine the most representative dilution necessary to detect a reduction in prevalence. An average of 300 samples for each type of meat were analyzed. According to Fisher's exact test, the dilution to be used to detect a reduction in prevalence was chosen based on an initial prevalence of 20 to 26%. Based on this introductory study, a new sampling plan representative of the nationwide Belgian meat production process was used from 2000 through to 2003. This study confirmed the consistently high rate and level of contamination of poultry meat: broiler and layer carcasses were the most contaminated samples followed by broiler fillets and poultry meat preparations. A constant and significant decrease in Salmonella prevalence was observed for pork carcasses, trimmings, and minced meat and for beef minced meat. Less than 3% of beef carcasses and trimming samples were positive for Salmonella. The Belgian plan, as utilized from 2000 to 2003, was suitable for monitoring of zoonoses because the sampling plan was representative of nationwide production processes, covered all periods of the year, and was executed by trained samplers and the analyses were carried out by recognized laboratories using an identical analytical method.