猪肉屠宰加工过程中弯曲菌流行病学调查与分析

目的 调查分析猪肉屠宰加工过程中主要环节弯曲菌的污染现状。方法 选择两家猪肉屠宰加工企业,分别采集宰前、脱毛、取内脏、胴体修饰和预冷前后等环节样品,应用选择性CCDA平板直接计数法对弯曲菌进行定量检测。结果 150份猪肉样品检测出22份弯曲菌阳性样品,阳性率14.67%;环境样品30份,检出12份阳性样品,阳性率40.00%。所检出的弯曲菌均为结肠弯曲菌,未见其它种属弯曲菌。所监测的6个环节均存在不同程度的弯曲菌污染,阳性率呈现先升高后降低的变化趋势。其中取内脏环节弯曲菌阳性率最高,为17.50%,阳性样品弯曲菌携带量为177.14±296.73CFU/100cm2;而环境样品中取内脏环节的地面样品阳性率最高,为100%,弯曲菌携带量为588.00±307.66CFU/100cm2。结论 猪肉屠宰加工过程中存在弯曲菌污染,以取内脏环节阳性率最高,预冷后阳性率明显降低;而环境样品的高污染率对猪肉屠宰加工过程中的交叉污染构成了很大威胁。     

黑龙江省肉鸡养殖和屠宰加工环节中沙门菌调查

目的了解黑龙江省肉鸡养殖和屠宰加工环节中沙门菌污染状况。方法根据国家食品安全风险监测专项《肉鸡养殖及屠宰加工环节沙门氏菌专项监测工作手册》规定的操作程序,并参考国家标准GB 4789.4—2010《食品安全国家标准食品微生物学检验沙门氏菌检验》对黑龙江省4个地市3 766份肉鸡样品进行检验。结果 4大环节共检测样品3 766份,检出阳性样品339份,检出率为9.00%,孵化、养殖、屠宰到配送分销4个环节均检出沙门菌,检出率分别为2.82%(22/781)、2.14%(11/515)、13.84%(220/1 590)、9.77%(86/880)。其中,屠宰环节的8种样品中预冷池水样品沙门菌检出率最高,为35.42%(34/96)。检出的沙门菌分属于9个血清型,以肠炎沙门菌和印地安纳沙门菌的比例最高。结论黑龙江省肉鸡屠宰和配送分销环节中沙门菌污染较严重。屠宰环节阳性率最高,是重要的污染环节,其中预冷池水中沙门菌检出率最高,是屠宰过程中肉鸡污染沙门菌的主要环节。     

肉鸡加工过程中沙门氏菌的污染监测及分析

目的对肉鸡屠宰加工的不同环节进行沙门氏菌监测,了解各个环节沙门氏菌的污染状况,分析肉鸡加工过程中沙门氏菌的污染来源及关键环节。方法选择河南省某大型肉鸡屠宰加工企业作为监测点,在肉鸡屠宰前、褪毛后、过预冷池后、被分割后和包装入库等环节及预冷池水、分割刀具案板和工人手等处分别采集样本,共计1756份,进行沙门氏菌检测及其血清型鉴定。结果于1756份样本中检出沙门氏菌阳性样本700份,检出率为39.9%。其中,褪毛后整禽的沙门氏菌检出率最高,达63.9%;其次为过预冷池后的整禽胴体,检出率为57.3%。屠宰加工环节中检出的沙门氏菌中肠炎沙门氏菌最多,占71.7%,其次为印第安纳沙门氏菌,占20.8%。结论肉鸡在养殖期间存在沙门氏菌污染,且在屠宰加工环节存在交叉污染,应针对相关环节进行重点控制和工艺改进。     

屠宰过程黄羽肉鸡弯曲菌的定量风险评估

目的 检测屠宰过程黄羽肉鸡弯曲菌阳性率和污染水平,建立屠宰过程黄羽肉鸡弯曲菌的定量风险评估模型,探明其污染的关键风险点。方法 用传统分离鉴定方法和最近似数法（most probable numbers,MPN）定量法对屠宰不同环节弯曲菌阳性率和污染水平进行检测,接着运用描述函数对数据进行拟合,并采用@Risk软件进行重复性检验和敏感性分析。结果 弯曲菌阳性率和污染水平以活鸡肛拭环节最高（53.60％,1.8300 logMPN/m2）,其次为打毛和开膛环节（27.50％,0.5000 logMPN/m2和18.10％,0.0270 logMPN/m2）。根据函数拟合结果建立弯曲菌污染定量风险评估模型,蒙特卡洛（Monte Carlo）模拟分析显示肛拭环节弯曲菌阳性率和污染水平最高(52.92%,1.8896 logMPN),其次是打毛和开膛环节(27.46%,0.4680 logMPN和14.74%,0.4910 logMPN),各个模拟环节数值与分离鉴定结果误差较小（P<0.05）,表明模型具有良好的准确性;敏感性分析获得黄羽肉鸡屠宰过程弯曲菌污染的关键风险点为打毛和开膛环节,相关系数分别为0.77和-0.39,高于其他环节。结论 本研究以屠宰过程黄羽肉鸡弯曲菌为研究对象进行风险评估,成功建立弯曲菌暴露评估模型,且模型具有较好的准确性,同时揭示了屠宰过程弯曲菌防控的关键风险点,为黄羽肉鸡弯曲菌的防控和风险评估奠定基础。     

2012—2013年成都市生鸡肉中弯曲菌调查分析

了解成都市零售环节鸡肉中的弯曲菌定量污染水平以及其随季节变化的趋势,为食品安全风险评估提供依据。方法 选择成都市及周边的大型超市和农贸市场作为采样点,采取未分割的新鲜鸡冷藏鸡胴体。用涂布平板法计数,用多重PCR进行鉴定。结果 采集255份样品,阳性数202份,检出率为79.22%;弯曲菌污染量＞500 cfu/g的样品占32.55%(83/255);农贸市场样品的阳性率高于超市样品;夏季阳性率高于其他季节;共分离到空肠弯曲菌300株、结肠弯曲菌393株和胎儿弯曲菌11株。结论 四川省零售环节鸡肉中弯曲菌的污染严重。     

肉鸡屠宰加工中减菌处理前后细菌菌相分析

基于16S rDNA V6～V8可变区的聚合酶链式反应-变性梯度凝胶电泳（polymerase chain reaction-denaturinggradient gel electrophoresis,PCR-DGGE）技术分析肉鸡屠宰加工过程中减菌处理前后胴体或产品细菌多样性。在预冷环节前采用50 ℃、1.5%乳酸溶液对肉鸡胴体冲淋15 s进行减菌处理,采集屠宰加工环节中减菌处理前后的胴体或分割产品表面样品,提取样品中的细菌总DNA,通过16S rDNA V6～V8可变区的PCR扩增,变性梯度凝胶电泳,对PCR扩增片段割胶回收、克隆测序分析减菌前后细菌菌相变化。结果表明,减菌前,胴体清洗环节DGGE条带的数量最多、亮度最强,细菌污染最严重,其次是分割环节,而预冷环节细菌种类及数量最少,污染程度最低;减菌后,各屠宰加工环节细菌种类与数量较减菌前均有所减少,其中胴体清洗环节与分割环节细菌的种类与数量减少量最多,预冷环节细菌的种类及数量最少,不同屠宰加工环节细菌种类并不完全一致;乳杆菌属细菌在整个肉鸡屠宰加工过程中均有出现,与肠杆菌科和假单胞菌属细菌为肉鸡屠宰加工过程中的优势腐败菌。     

环介导等温扩增法快速检测感染性腹泻病例标本中的空肠弯曲菌

目的 建立环介导等温扩增法(loop-mediated isothermal amplification, LAMP)快速检测感染性腹泻病例标本中的空肠弯曲菌。方法 收集2017年苏州市辖区3家医院的感染性腹泻患者的粪便样品326件, 用LAMP法对空肠弯曲菌的DNA进行检测, 在DNA扩增试剂盒提供的反应体系加入引物和模板进行空肠弯曲菌的LAMP反应, 测定LAMP方法检测空肠弯曲菌菌液的灵敏度; 同时用滤膜法进行空肠弯曲菌的分离培养, 参照GB 4789.9-2014《食品安全国家标准 食品微生物学检验 空肠弯曲菌检验》分离鉴定空肠弯曲菌。结果 LAMP法检测空肠弯曲菌菌液的灵敏度是2.2 CFU/μL; 326件腹泻患者粪便中16件空肠弯曲菌LAMP检测为阳性, 阳性率为4.91%。共分离培养出14株空肠弯曲菌, 阳性率为4.29%。结论 针对感染性腹泻患者的粪便样本的空肠弯曲菌检测项目, 可以采用LAMP方法进行等温扩增初筛, 初筛阳性的样品再有针对性地进行后续的传统培养。     

黄羽肉鸡屠宰过程中胴体表面腐败菌的变化

为了分析黄羽肉鸡屠宰加工环节的肉鸡胴体污染菌的菌群组成,本试验利用平板倾注法及Illumina MiSeq高通量测序高效地测定黄羽肉鸡屠宰过程中加工环境和胴体表面腐败菌的多样性。结果表明,净膛、预冷及分级是黄羽肉鸡菌落总数增长的主要污染来源工序,分级秤、分级车间工人手套、预冷槽及净膛工人手套为以上三个工序中污染来源接触面,且分级秤及分级车间工人手套所污染菌群是黄羽鸡胴体菌群的主要来源,使黄羽鸡胴体表面菌落总数及假单胞菌增长率高达24.13%、41.27%,经过分级秤及分级车间工人手后黄羽鸡菌落总数显著（P<0.05）增加至4.63 lg(CFU/g)。打毛后（DM）、净膛后（JH）、净膛消毒后（CH）黄羽鸡胴体优势菌主要为链球菌属（Streptococcus）,大肠杆菌属（Escherichia）和气单胞菌属（Aeromonas）。黄羽鸡经预冷槽预冷后气单胞菌属丰度大幅增加,链球菌属次之。经过分级秤分级后黄羽鸡胴体菌群中不动杆菌属（Acinetobacter）为主要优势菌,巨球菌属（Macrococcus）次之。添加次氯酸电解水减菌后,黄羽肉鸡胴体表明气单胞菌属和链球菌属丰度大幅...     

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。     

肉鸡屠宰环节金黄色葡萄球菌污染定量评估研究

目的解析某大型肉鸡屠宰场中金黄色葡萄球菌的风险和关键防控点,为肉鸡屠宰过程金黄色葡萄球菌污染的科学防控提供指导。方法通过某大型肉鸡屠宰场金黄色葡萄球菌的监测和调研,应用@RISK 7软件建立金黄色葡萄球菌污染的定量评估模型,对肉鸡屠宰四个环节(烫洗煺毛、净膛、清洗预冷和分割传送)进行定量评估。结果初步探明了鸡肉中金黄色葡萄球菌在屠宰加工过程中的消长变化规律,清洗预冷环节和分割传送环节是金黄色葡萄球菌污染的主要风险环节,预冷池水中金黄色葡萄球菌浓度和工人手部带菌程度是肉鸡屠宰加工过程中金黄色葡萄球菌的关键风险点(相关系数分别为0.62和0.50)。结论对肉鸡屠宰环节的金黄色葡萄球菌进行关键控制点识别和精准采取控制措施,可有效保障终端鸡肉产品的卫生安全。     

Incidence of Salmonella, Campylobacter jejuni, Campylobacter coli, and Listeria monocytogenes in poultry carcasses and different types of poultry products for sale on the Belgian retail market.

From January 1997 to May 1998, 772 samples of poultry carcasses and poultry products for sale on the retail market in Belgium were analyzed for the presence of Salmonella spp., Salmonella Enteritidis, Campylobacter jejuni, C. coli, and Listeria monocytogenes per 100 cm2 or 25 g. Poultry samples were contaminated with Salmonella (36.5%), C. jejuni and C. coli (28.5%), and L. monocytogenes (38.2%). In about 12.3% of the poultry samples, the L. monocytogenes contamination level exceeded 1 CFU per g or cm2. Significant differences in pathogen contamination rates of poultry products were noticed between the poultry products originating from Belgian, French, and U.K. abattoirs. Poultry products derived from broiler chickens running free in pine woods until slaughtering age (12 to 13 weeks) had a significantly (P < 0.05) lower contamination rate of Salmonella than poultry products from enclosed broilers slaughtered at the age of 6 to 8 weeks. A significantly (P < 0.05) lower pathogen contamination rate was noted for Salmonella, C. jejuni, and C. coli for poultry cuts without skin compared to poultry cuts with skin on. An increase in pathogen contamination rate was noticed during cutting and further processing. To diminish C. jejuni, C. coli, Salmonella, and L. monocytogenes contamination rates, hygienic rules of slaughter and meat processing must be rigorously observed. At the moment, zero tolerance for these pathogens is not feasible, and there is a need to establish criteria allowing these pathogens to be present at reasonable levels in the examined poultry samples.     

Contamination of meat with Campylobacter jejuni in Saitama, Japan.

To determine the source of food contamination with Campylobacter jejuni, we investigated retail meat, a chicken processing plant and a broiler farm. C. jejuni was found in domestic retailed poultry (45.8%) and imported poultry (3.7%), but not in beef or pork. In the poultry processing plant, there is significant contamination with C. jejuni in chicken carcasses, equipment and workers' hands. This contamination increases during the defeathering and evisceration processes. RAPD analysis shows that contamination with C. jejuni is of intestinal origin. In a broiler farm, C. jejuni was first isolated from a faecal sample of broiler chicken after the 20th day of age. Two weeks later, all birds in this farm became C. jejuni positive. RAPD analysis indicated that C. jejuni spread rapidly from one broiler flock to the other flocks on the farm.     

陕西省某活鸡屠宰场肉鸡相关样品中沙门菌定量检测及污染分析

目的了解肉鸡屠宰加工中不同时间和环节沙门菌的污染情况,分析污染关键点。方法 2016年11月至2017年11月从陕西省某活鸡屠宰场不同环节定期采集活鸡肛拭子标本、整鸡胴体和鸡肉样品,使用最大可能数(MPN)法对沙门菌进行定量检测,同时分离菌株;采用聚合酶链式反应(PCR)技术对沙门菌进行鉴定,同时结合血清凝集技术对沙门菌鉴定结果进行确认。结果采集的284份样品中有67份检出沙门菌,检出率为23. 6%,平均MPN值为0. 051 6 MPN/g。2017年7月采集的样品沙门菌污染最为严重,检出率为37. 8%(14/37),平均MPN值为0. 064 7 MPN/g; 2016年11月检出率最低,为13. 9%(5/36),平均MPN值为0. 043 6 MPN/g。不同屠宰环节中,浸烫褪毛后整鸡胴体样品中沙门菌检出率最高(43. 3%,26/60),平均MPN值为0. 060 5 MPN/g;分割后冷冻前鸡胸脯肉样品中沙门菌检出率最低(18. 3%,11/60),平均MPN值为0. 036 8 MPN/g,略高于储存配送过程整鸡胴体/鸡胸脯肉样品中沙门菌的污染水平(0. 035 8 MPN/g)。结论活鸡屠宰过程沙门菌的检出率与MPN值具有较强的季节性,在不同屠宰加工环节存在纵向和交叉污染,应对活鸡屠宰加工过程沙门菌污染严重的环节进行重点控制。     

Prevalence of Listeria monocytogenes in broilers at the abattoir, processing plant, and retail level.

The environment and products from two broiler abattoirs and processing plants and raw broiler pieces at the retail level were sampled for Listeria monocytogenes in order to evaluate the contamination level of the broiler carcasses and products. Sampling started in the slaughtering process and finished with raw broiler meat or ready-to-eat cooked product. Sampling sites positive for L. monocytogenes at the broiler abattoir were the air chiller, the skin-removing machine, and the conveyor belt leading to the packaging area. The L monocytogenes contamination rate varied from 1 to 19% between the two plants studied. Furthermore, 62% (38 of 61) of the raw broiler pieces, bought from retail stores, were positive for L. monocytogenes. Altogether, 136 L. monocytogenes isolates were obtained for serotyping and pulsed-field gel electrophoresis (PFGE) characterization performed with two rare-cutting enzymes (ApaI and AscI). Altogether three serotypes (1/2a, 1/2c, and 4b) and 14 different PFGE types were obtained using information provided from both ApaI and AscI patterns for discrimination basis. The two broiler abattoirs studied did not share the same PFGE types. However, the same PFGE types found in the raw broiler pieces at the retail level were also found in the broiler abattoirs where the broilers had been slaughtered.     

ENUMERATION OF AEROMONAS FOR VERIFICATION OF THE HYGIENIC ADEQUACY OF SWINE CARCASS DRESSING PROCESSES1

Populations of Aeromonas spp. and aerobic bacteria from dehairing equipment and from carcasses passing through different processing steps in a swine slaughtering plant were evaluated to identify the hygienic risks of each operation. Sponge samples were taken from the scraper flails in dehairing machines and the surface of the shackling table at pre‐ and post‐operation times, with 20 samples from each location being collected at each time. Increased post‐operation levels of Aeromonas spp. indicated a buildup and possible spread of these bacteria to carcasses. The belly skins of 40 dehaired carcasses were each sampled at five points along the process line which were after the shackling, after the final singeing, after the final polishing, after the final wash and after chilling. The levels of microbial contaminants on carcasses varied at each processing step. The heaviest contamination of carcasses with Aeromonas (1.88 log CFU/cm²) and aerobic bacteria (2.66 log CFU/cm²) occurred after shackling. Counts were reduced at other steps as a result of singeing, washing and chilling operations. However, singed carcasses were recontaminated with Aeromonas and aerobic bacteria during the polishing operation. Aeromonas hydrophila were the most prominent motile aeromonads (74.1%) recovered at the plant. The findings for Aeromonas spp. were similar to those for aerobic bacteria (r²= 0.9995) which suggested that Aeromonas spp. are appropriate indicators for assessing carcass dressing processes.     

Prevalence of Campylobacter, Salmonella, and Escherichia coli on the external packaging of raw meat

During September and October 2002, 3,662 prepackaged raw meat samples were collected to evaluate the extent and nature of microbiological contamination on external surfaces of the packaging, which could potentially cross-contaminate ready-to-eat foods during and after purchase. Salmonella was detected on two (<1%) samples of external packaging (both from raw chicken), and Campylobacter was detected on 41 (1.1%) samples of external packaging. The external packaging of game fowl exhibited the highest Campylobacter contamination (3.6%), followed by raw chicken (3.0%), lamb (1.6%), turkey (0.8%), pork (0.2%), and beef (0.1%); Campylobacter jejuni and Campylobacter coli accounted for 59% (24 of 41) and 24% (10 of 41) of the contaminating Campylobacter species, respectively. C. coli isolates from the external packaging were more multiresistant to antimicrobial drugs, including quinolones such as ciprofloxacin, than was C. jejuni. Escherichia coli (an indicator of fecal contamination) was isolated from the external packaging on 4% of the raw meat samples at levels of 40 to 10(5) CFU per swab. The external packaging of raw meats is a vehicle for potential cross-contamination by Campylobacter, Salmonella, and E. coli in retail premises and consumers' homes. The external surface of heat-sealed packaging was less frequently contaminated with Campylobacter and E. coli compared with other types of packaging (e.g., overwrapping, bag, and tie tape) (P < 0.0001 to 0.01). In addition, external packaging of raw meats was contaminated less frequently with Campylobacter and E. coli when packaging was intact, packaging and display areas were visually clean, display temperatures were below 8 degrees C, and hazard analysis systems were in place.     

Effect of chemical dehairing on the prevalence of Escherichia coli O157:H7 and the levels of aerobic bacteria and enterobacteriaceae on carcasses in a commercial beef processing plant

The objective of this experiment was to test the hypothesis that cleaning cattle hides by removing hair and extraneous matter before hide removal would result in improved microbiological quality of carcasses in commercial beef processing plants. To test this hypothesis, we examined the effect of chemical dehairing of cattle hides on the prevalence of Escherichia coli O157:H7 and the levels of aerobic bacteria and Enterobacteriaceae on carcasses. Samples from 240 control (conventionally processed) and 240 treated (chemically dehaired before hide removal) hides (immediately after stunning but before treatment) and preevisceration carcasses (immediately after hide removal) were obtained from four visits to a commercial beef processing plant. Total aerobic plate counts (APC) and Enterobacteriaceae counts (EBC) were not (P > 0.05) different between cattle designated for chemical dehairing (8.1 and 5.9 log CFU/100 cm2 for APC and EBC, respectively) and cattle designated for conventional processing (8.0 and 5.7 log CFU/100 cm2 for APC and EBC, respectively). However, E. coli O157:H7 hide prevalence was higher (P < 0.05) for the control group than for the treated group (67% versus 88%). In contrast to hides, the bacterial levels were lower (P < 0.05) on the treated (3.5 and 1.4 log CFU/100 cm2 for APC and EBC) than the control (5.5 and 3.2 log CFU/100 cm2 for APC and EBC) preevisceration carcasses. Prevalence of E. coli O157:H7 was lower (P > 0.05) on treated than on control preevisceration carcasses (1% versus 50%). These data indicate that chemical dehairing of cattle hides is an effective intervention to reduce the incidence of hide-to-carcass contamination with pathogens. The data also imply that any effective hide intervention process incorporated into beef processing procedures would significantly reduce carcass contamination by E. coli O157:H7.     

Occurrence of Listeria monocytogenes in freshwater fish farms and fish-smoking plants

Three Swiss fish farms, farming rainbow trout (Oncorhynchus mykiss), and their affiliated smoking plants were analyzed for the presence of Listeria spp. 590 samples were collected from the farming environment (raceway water, sludge), faecal content and skin of the fish, fish during processing, and the processing environment.Listeria spp. were found at prevalences of 2·3% in plant A, 31·6% in plant B (mainly L. monocytogenes), and 13·8% in plant C (mainly L. innocua). This high contamination rate in plant B may be explained by the following facts: (i) farm B uses river water flowing through agricultural land; (ii) plant B rears fish in earth ponds instead of concrete ponds or raceways; (iii) fish from farm B had not been denied feed prior to slaughter; and (iv) total lack of regular mechanical and chemical cleaning in the fish farm B and processing plant B.In all three plants samples taken after smoking but before packaging did not contain Listeria spp., although in plant B and C the raw fish was contaminated. Hygienic defaults during packaging can lead to contaminated ready-to-eat products, detected in plant B (L. monocytogenes) and plant C (L. innocua) with one sample each. To minimize a possible health hazard to the consumer, it is of great importance to prevent postprocessing contamination of smoked fish.Finally, means of preventing Listeria contamination during farming, slaughtering, processing and storage are suggested.