3M PetrifilmTM环境李斯特菌测试片的比较实验

3M Petrifilm TM环境李斯特菌测试片法是一种检测工厂环境中李斯特菌属的快速检测方法,主要针对食品生产车间中食品接触表面(传送带、切片机、托盘等)采样,非食品接触表面(地板、桌面、排水口等)采样和地下水的检测,是企业切实加强对单核增生李斯特菌控制的有力手段。本实验用3M Petrifilm TM环境李斯特     

3M PetrifiLm~(TM)环境李斯特菌测试片的比较实验

<正>3M Petrifilm TM环境李斯特菌测试片法是一种检测工厂环境中李斯特菌属的快速检测方法,主要针对食品生产车间中食品接触表面(传送带、切片机、托盘等)采样,非食品接触表面(地板、桌面、排水口等)采样和地下水的检测,是企业切实加强对单核增生李斯特菌控制的有力手段。本实验用3M Petrifilm TM环境李斯特     

单增李斯特菌便捷检测试纸的制备及检测效果

目的：研究一种快速、便捷检测食品中单增李斯特菌的方法,拟对本实验室制备的单增李斯特菌快速检测试纸（Listeria monocytogenes-fast test paper,LM-FTP）（专利受理号：201510274726.9）进行检测效果评价。方法：采集肉样、乳样、水样和蔬菜,用LM-FTP进行检测,并与市售单增李斯特菌检测试纸片（3M PetrifilmTM）进行比对实验。结果：LM-FTP的检测特异性为100%,检测限为2.1×104 CFU/mL。增菌培养到报告结果只需5.5 h。LM-FTP和3M PetrifilmTM检测都可对样品完成检测,对农贸市场畜产品及蔬菜检测,二者符合率为75%,对超市同类食品检测,二者符合率为62.5%;对超市熟食检测时,二者符合率为93.75%。结论：与市售3M PetrifilmTM检测方法相比,LM-FTP检测试纸制备简单,操作便捷,较3M PetrifilmTM的24 h报告结果更节省时间,但在检测灵敏度方面有一定差距,尚需进一步研究改进。     

食品中菌落总数检测方法的比较研究

本文比较了大肠埃希氏菌、金黄色葡萄球菌和蜡样芽孢杆菌3种标准菌株分别在平板计数琼脂、3M菌落总数测试片和MicroFast~?AC菌落总数测试片3种培养基中的菌落形态,并采用这3种方法对6类共60批次食品样品进行菌落总数测定。结果表明,3M菌落总数测试片在菌落辨识度上有明显优势,更适用于复杂基质样品的检测;3M菌落总数测试片和MicroFast~?AC菌落总数测试片测得的结果与平板计数法测得的结果无显著差异,菌落总数测试片法操作简单,节省空间,但成本较高,食品检测机构和相关企业可根据实际工作情况选择检测方法。     

沙门氏菌快速测试片在食品检测中的初步应用研究

目的应用3M沙门氏菌SALX测试片法检测食品中的沙门氏菌。方法对67批自然样品,以及使用3个沙门氏菌标准菌株进行人工污染的20批样品,同时采用3M沙门氏菌SALX测试片法与国家标准方法GB 4789.4-2010进行检测并比较,评价3M SALX沙门氏菌测试片法检测沙门氏菌的检测性能。结果 3M沙门氏菌SALX测试片法与国标法符合率达95.5%,各有1例假阳性和假阴性。结论 3M沙门氏菌SALX测试片法操作相对简便,稳定性好,与国标法有较高的符合率,作为一种新的沙门氏菌检测方法,仍需进一步积累检测数据。     

3M PetrifilmTM快速菌落总数测试片的评价

在食品菌落总数的检测中,评估3M PetrifilmTM快速菌落总数测试片的准确性和适用性.以国家标准GB 4789.2—2016为参考方法,测试5类15种具有代表性的食品,评估测试片方法的线性和准确度;同时评估测试片的耐变性和批间变异;并开展实验室间的协同实验.结果表明,3M PetrifilmTM快速菌落总数测试片...     

测试片法在食品菌落总数检测中的应用研究

目的 探究菌落总数测试片法检测食品中菌落总数的可行性.方法 用菌落总数测试片法和国家标准方法同时对制备的菌悬液、自然污染食品样品和人工污染食品样品的菌落总数进行检测,采用t-检验分析2种方法检测结果的差异,采用Pearson相关分析确定2种检测方法之间的相关性.结果 菌悬液和自然污染食品样品菌落总数测试片法的变异系数为...     

散装熟肉制品中单增李斯特菌污染状况分析

目的分析散装熟肉制品中单增李斯特菌污染状况。方法根据GB 4789.30-2016《单核细胞增生李斯特氏菌检验》规定的方法对散装熟肉制品的加工用原料、生产环境、各加工环节中产品以及不同销售环境下产品中的单增李斯特菌进行定性和定量检测。结果原料肉的总体带菌率达到21%;生产环境中第三区(远离食品接触面的区域)检出率为2%,其余区域未检出;各加工环节中产品单增李斯特菌检测结果均小于10CFU/g;不同销售环境下产品中单增李斯特菌检测结果小于10 CFU/g的比例为93%,处于10～50 CFU/g之间的样本占比6%,大于50 CFU/g的占比为1%。结论原料散装熟肉制品中单增李斯特菌污染的主要来源,蒸煮等加工环节能有效地杀灭单增李斯特菌。销售环境也关系到散装熟肉制品单增李斯特菌的污染程度,对于未包装的产品,专卖店优于农产品市场。     

工厂中环境和海产品加工前后的微生物学检验

检测了2个海产品加工厂的环境及加工前后海产品样品的菌落总数(个/cm2,个/g或个/mL)、李斯特氏菌和单核细胞增生李氏菌数(每100cm2,100g或100mL样品的MPN数),其结果如下:2个工厂对应样品的菌落总数及其分布规律相接近,但李斯特氏菌和单核细胞增生李氏菌的出现率和数量却相差很大;单核细胞增生李氏菌数都低于或远低于李斯特氏菌,且单核细胞增生李氏菌与李斯特氏菌的数量和分布呈正相关;加工后海产品中的李斯特氏菌和单核细胞增生李氏菌可能来自原料和加工过程;环境中李斯特氏菌和单核细胞增生李氏菌主要来自原料.     

预包装熟肉制品加工环境李斯特菌监控方案的构建

单核细胞增生李斯特氏菌作为一种食源性致病菌,该菌可存在于各类食品中,尤其是即食类食品,也可在食品加工环境包括生产设备、用具、地板和排水沟等中长期生存。虽然发病率相对于其他食源性致病菌较低,但危害较大。因此,对加工环境中的单核细胞增生李斯特氏菌的污染控制尤为重要。加工环境中李斯特菌监控作为一种预警和验证手段,可以在终产品检出单核细胞增生李斯特氏菌之前做好预防措施。本文汇总分析了多个加工肉制品企业环境监测的经验,构建出肉制品加工过程环境中李斯特菌监控基础方案,包括较为详细的区域划分、采样分区、采样及检测方法、预防措施和纠正措施等,形成适合自身企业的环境李斯特菌监控方案,为肉制品加工行业卫生规范细化提供一定参考依据。     

Comparison of 3M Petrifilm environmental Listeria plates against standard enrichment methods for the detection of Listeria monocytogenes of epidemiological significance from environmental surfaces

ABSTRACT:  Environmental monitoring using sensitive methods for detection and elimination of harborage sites of Listeria monocytogenes is key to the control of this organism. The 3M™ Petrifilm™ Environmental Listeria (EL) Plate—a no enrichment method—was compared with the USDA/FSIS, modified USDA/FSIS (mUSDA), and ISO methods for detection/recovery of L. monocytogenes on 4 environmental surfaces (brick, dairy board, stainless steel, and epoxy resin). The efficacy of 3 sampling devices including the Microbial-Vac system^®, environmental sponge, and 3M Quick swab in recovering epidemiologically significant strains of uninjured and sublethally injured L. monocytogenes from environmental surfaces was evaluated. Environmental surfaces were inoculated with Listeria to obtain final cell densities of approximately 10 to 100 CFU/100 cm². The surfaces were then sampled and processed. For all methods, percent recovery (% samples where Listeria was detected) was significantly higher ( P < 0.05) for uninjured cells (75% to 100%) compared to injured cells (58.9% to 81.1%). The Petrifilm EL Plate method efficiently recovered both low level and injured Listeria populations from environmental test surfaces when used in conjunction with environmental sponge and the 3M Quick swab sampling. The mUSDA method was superior to all other methods for recovering both uninjured (100% recovery) and injured L. monocytogenes (80.8% to 81.1% recovery). Sponges and swabs were equally effective in recovering uninjured and injured Listeria and were significantly different ( P < 0.05) from the Microbial-Vac system. The findings indicate that both mUSDA and Petrifilm EL Plate methods can be used for the detection of potentially injured Listeria on food processing environmental surfaces.     

Peptide nucleic acid fluorescence in situ hybridization for identification of Listeria genus, Listeria monocytogenes and Listeria ivanovii

A fluorescent in situ hybridization (FISH) method in conjunction with fluorescin-labeled peptide nucleic acid (PNA) probes (PNA-FISH) for detection of Listeria species was developed. In silico analysis showed that three PNA probes Lis-16S-1, Lm-16S-2 and Liv-16S-5 were suitable for specific identification of Listeria genus, Listeria monocytogenes and Listeria ivanovii, respectively. These probes were experimentally verified by their reactivity against 19 strains of six Listeria species (excluding newly described species Listeria marthii and Listeria rocourtiae) and eight other bacterial species. The PNA-FISH method was optimized as 30 min of hybridization with 0.2% Triton X-100 in the solution and used to identify 85 Listeria strains from individual putative Listeria colonies on PALCAM agar plates streaked from selectively enriched cultures of 780 food or food-related samples. Of the 85 Listeria strains, thirty-seven were identified as L. monocytogenes with the probe Lm-16S-2 and two as L. ivanovii with the probe Liv-16S-5 which was in agreement with the results obtained by the API LISTERIA method. Thus, the PNA-FISH protocol has the potential for identification of pathogenic Listeria spp. from food or food-related samples.     

Evaluation of the University of Minnesota Tri-plate and 3M Petrifilm for the isolation of Staphylococcus aureus and Streptococcus species from clinically mastitic milk samples

The primary objective was to compare microbiological results of the University of Minnesota Tri-plate and the 3M Petrifilm Staph Express (STX) Count Plate to standard culture techniques for identification of clinical mastitis caused by Staphylococcus aureus. The secondary objective was to evaluate the Tri-plate's ability to differentiate Streptococcus spp. from other gram-positive organisms. The tests were evaluated using clinically positive mastitic milk samples (n = 282) to determine their ability to diagnose the pathogens of interest. A Tri-plate was classified positive for Staph. aureus when at least 1 colony exhibiting β-hemolysis was present on the Factor media portion of the plate. When the plate was used in this manner and read by a trained laboratory technician, the sensitivity of the Tri-plate was 97.9% and the specificity was 81.8%. When the Tri-plate was evaluated by the laboratory technician for its ability to diagnose Streptococcus spp., both sensitivity and specificity of the test were very good (92.6 and 89.5%, respectively). Using the Petrifilm, samples were classified as positive for Staph. aureus if any red-violet colonies were present on the Petrifilm after an initial 24-h incubation. When used in this manner, the Petrifilm had a sensitivity of 97.4% and a specificity of 76.1%. Further evaluation of the Petrifilm was done using the STX disk, which was used to confirm the presence of Staph. aureus. When using the presence of 1 pink colony on the disk, the sensitivity of the Petrifilm was 92.1% and the specificity was 93.1%. Both the Tri-plate and the 3M STX Petrifilm successfully diagnosed Staph. aureus in clinical milk samples when used in a laboratory setting and the Tri-plate successfully differentiated Streptococcus spp. from other gram-positive organisms.     

Petrifilm--up-to-date tests for microbiological control of food products, raw materials and environmental objects

The principle of action, advantages and different types of Petrifilm as alternative test to the classical microbiological analysis are considered. Petrifilm is up-to-date high technology test systems for the rapid quantitative microbiological control in foodstuff of sanitary-significant and pathogenic microorganisms--yeast and moulds, coliforms, Enterobacteriaceae, Staphylococcus, Listeria spp. and Lactobacilli spp. Automatic testing of colonies on Petrifilm with use of the Petrifilm-Reader allows to reduce time for colony counting, to exclude errors of personnel and to document results of the analysis.     

A new rapid automated method for the detection of Listeria from environmental swabs and sponges

Many food and meat processors test environmental swabs and sponges to confirm the absence of Listeria spp. Spectral pattern changes in a liquid growth medium, resulting from esculin hydrolysis by Listeria in contaminated swabs and sponges, were automatically monitored by the BioSys instrument in a semifluid layer (SFL). The blackening of SFL in modified MOX broth resulted in sharply declining curves, which were easily detected by the instrument. The instrument detected all nine strains of Listeria monocytogenes tested. None of the gram negative organisms (Proteus, Escherichia coli, Pseudomonas, Citrobacter and Yersinia) were detected by the system, nor were most gram positive organisms, including Bacillus, Streptococcus, and Lactobacillus strains, Staphylococcus aureus, Enterococcus faecium and E. faecalis, which hydrolyze esculin, produced black colonies on PALCAM and Oxford media and were also detected in the system. A total of 122 sponges and swabs collected at food processing plants were evaluated by this method. Of these, 99 were negative, and 11 were positive. L. innocua was the dominant Listeria species in these environmental samples. Good correlation was obtained between numbers of Listeria and detection times of esculin hydrolysis: 1000 CFU/swab were detected in 10-13 h, whereas 1-10 CFU/swab were detected in less than 22 h. The total assay time was 26 h.     

Tracking of Listeria monocytogenes in smoked fish processing plants

Four smoked fish processing plants were used as a model system to characterize Listeria monocytogenes contamination patterns in ready-to-eat food production environments. Each of the four plants was sampled monthly for approximately 1 year. At each sampling, four to six raw fish and four to six finished product samples were collected from corresponding lots. In addition, 12 to 14 environmental sponge samples were collected several hours after the start of production at sites selected as being likely contamination sources. A total of 234 raw fish, 233 finished products, and 553 environmental samples were tested. Presumptive Listeria spp. were isolated from 16.7% of the raw fish samples, 9.0% of the finished product samples, and 27.3% of the environmental samples. L. monocytogenes was isolated from 3.8% of the raw fish samples (0 to 10%, depending on the plant), 1.3% of the finished product samples (0 to 3.3%), and 12.8% of the environmental samples (0 to 29.8%). Among the environmental samples, L. monocytogenes was found in 23.7% of the samples taken from drains, 4.8% of the samples taken from food contact surfaces, 10.4% of the samples taken from employee contact surfaces (aprons, hands, and door handles), and 12.3% of the samples taken from other nonfood contact surfaces. Listeria spp. were isolated from environmental samples in each of the four plants, whereas L. monocytogenes was not found in any of the environmental samples from one plant. Overall, the L. monocytogenes prevalence in the plant environment showed a statistically significant (P < 0.0001) positive relationship with the prevalence of this organism in finished product samples. Automated EcoRI ribotyping differentiated 15 ribotypes among the 83 L. monocytogenes isolates. For each of the three plants with L. monocytogenes-positive environmental samples, one or two ribotypes seemed to persist in the plant environment during the study period. In one plant, a specific L. monocytogenes ribotype represented 44% of the L. monocytogenes-positive environmental samples and was also responsible for one of the two finished product positives found in this plant. In another plant, a specific L. monocytogenes ribotype persisted in the raw fish handling area. However, this ribotype was never isolated from the finished product area in this plant, indicating that this operation has achieved effective separation of raw and finished product areas. Molecular subtyping methods can help identify plant-specific L. monocytogenes contamination routes and thus provide the knowledge needed to implement improved L. monocytogenes control strategies.     

Longitudinal studies on Listeria in smoked fish plants: impact of intervention strategies on contamination patterns

Four ready-to-eat smoked fish plants were monitored for 2 years to study Listeria contamination patterns and the impact of plant-specific Listeria control strategies, including employee training and targeted sanitation procedures, on Listeria contamination patterns. Samples from the processing plant environment and from raw and finished product were collected monthly and tested for Listeria spp. and Listeria monocytogenes. Before implementation of intervention strategies, 19.2% of raw product samples (n = 276), 8.7% of finished product samples (n = 275), and 26.1% of environmental samples (n = 617) tested positive for Listeria spp. During and after implementation of Listeria control strategies, 19.0% of raw product samples (n = 242), 7.0% of finished product samples (n = 244), and 19.5% of environmental samples (n = 527) were positive for Listeria spp. In one of the four fish plants (plant 4), no environmental samples were positive for L. monocytogenes, and this plant was thus excluded from statistical analyses. Based on data pooled from plants 1, 2, and 3, environmental Listeria spp. prevalence was significantly lower (P < 0.05) for nonfood contact surfaces and the finished product area and for the overall core environmental samples after implementation of control strategies. Listeria prevalence for floor drains was similar before and after implementation of controls (49.6 and 54.2%, respectively). Regression analysis revealed a significant positive relationship (P < 0.05) between L. monocytogenes prevalence in the environment and in finished products before implementation of control strategies; however, this relationship was absolved by implementation of Listeria control strategies. Molecular subtyping (EcoRI ribotyping) revealed that specific L. monocytogenes ribotypes persisted in three processing plants over time. These persistent ribotypes were responsible for all six finished product contamination events detected in plant 1. Ribotype data also indicated that incoming raw material is only rarely a direct source of finished product contamination. While these data indicate that plant-specific Listeria control strategies can reduce cross-contamination and prevalence of Listeria spp. and L. monocytogenes in the plant environment, elimination of persistent L. monocytogenes strains remains a considerable challenge.     

Evaluation of the 3M Petrifilm Enterobacteriaceae Count plate method for the enumeration of Enterobacteriaceae in foods

Results of the 3M Petrifilm Enterobacteriaceae Count (EB) plate method were compared with those of the standard violet red bile glucose agar (VRBG) method for the detection and enumeration of Enterobacteriaceae. Studies involving 107 bacterial strains demonstrated that the Petrifilm EB plate method is as sensitive as and more selective than the VRBG method. Sixty of the 62 pure Enterobacteriaceae cultures were recovered by both methods. In addition, 38 of the 45 non-Enterobacteriaceae organisms did not grow on the Petrifilm EB plate, while 28 of the 45 non-Enterobacteriaceae organisms did not grow on the VRBG plate. Colony counts from 174 naturally contaminated and 120 artificially inoculated dairy and nondairy food samples showed that the Petrifilm EB plate method performed as well as or better than the standard VRBG method for the enumeration of Enterobacteriaceae.     

南京市某食品厂酱卤熟肉制品生产加工过程微生物污染状况分析

目的了解南京市某食品厂酱卤熟肉制品加工过程中微生物污染状况及分布情况。方法 2016—2017年在南京市某食品厂采集生产加工过程中食品样品120份(原辅料60份、中间产品28份、成品24份、终产品8份)、环境样品204份、空气样品58份和生产用水14份,共396份,按照GB 4789食品微生物学检验系列标准和GB/T 16294—2010《医药工业洁净室(区)沉降菌的测试方法》对卫生指示菌和主要食源性致病菌进行检测,并对沙门菌进行血清型鉴定。结果在食品样品和环境样品中,李斯特菌检出率为7.4%(24/324),金黄色葡萄球菌检出率为2.6%(5/196),沙门菌检出率为1.9%(6/324);其中李斯特菌有2株单核细胞增生李斯特菌,其他以格氏李斯特菌和伊氏李斯特菌为主;沙门菌经血清学分型共4种血清型,分别为肠炎沙门菌(2株)、鼠伤寒沙门菌(2株)、依桑吉沙门菌(1株)和亚利桑那沙门菌(1株)。在原辅料、中间产品以及人员、仪器设备、清洁工具中均有致病菌检出,但成品和终产品中未检出致病菌。结论酱卤熟肉制品生产过程原辅料和环境中均存在多种致病菌污染。