2009—2012年上海市副溶血性弧菌血清分型研究

目的 了解2009-2012年期间上海市副溶血性弧菌临床菌株和水产品分离株的血清型分布.方法 采用血清玻片凝集试验对622株临床菌株和176株水产品分离株进行血清分型.结果 622株副溶血性弧菌临床菌株可分为27个血清型,其中以O3∶ K6型为主,约占61.7％.176株副溶血性弧菌水产品分离株可分为9个O群,其中O1、O2、O3、O4和O5群为主要血清型,约占72.2％.水产品分离株与临床菌株的血清型差异较大,但在水产品中检测到2株临床菌株的优势血清型O3∶ K6.结论 2009-2012年期间上海市副溶血性孤菌临床菌株和水产品分离株的血清型均呈现高度多样性,O3∶ K6型是引起食物中毒的优势血清型,而水产品分离株未见优势血清型.临床菌株优势血清型O3∶K6在水产品中的出现,提示上海市水产品存在潜在的致病危险,需要加强监控,为疾病的预防和控制提供依据.     

151株副溶血性弧菌的分子分型研究

目的 了解辽宁省副溶血性弧菌的分布规律及流行趋势，为辽宁省副溶血性弧菌引起的食源性疾病的溯源和预警提供基础。方法 对2019年辽宁省151株VP分离株进行血清分型、PFGE、REP-PCR和ERIC-PCR分子分型和聚类分析，评价3种方法间的关联性，同时探讨菌株间的亲缘关系。结果 124株分离株可共分为18个血清型，27株分离株未能分型，血清群以O3群、O1群和O2群为主。PFGE的分辨力（DI）为0.97,REP-PCR的DI为0.95,ERIC-PCR的DI为0.93。血清型O3群菌株与O1群菌株分子型相似度高。结论 2019年辽宁省临床VP分离株流行血清型仍为O3:K6，食品VP分离株流行血清群仍为O2。PFGE、REP-PCR和ERIC-PCR 3种分型方法结果一致，且PFGE分型方法的分辨力和再现性均优于其他两种方法，研究结果可以为副溶血性弧菌所引起食源性疾病的溯源提供新的技术手段。     

2020年辽宁省副溶血性弧菌PFGE分型、血清型分布和耐药谱分析

目的 了解辽宁省副溶血性弧菌（VP）的流行特征,为食源性疾病的防控提供可靠的技术支持,提高对食物中毒应急事件的处理能力。方法 对2020年从辽宁省内14个市内食源性疾病病人和食品中分离的VP进行PFGE分子分型、血清学分型、药物敏感试验并分析。结果 44株VP（食源性疾病分离株22株,食品分离株22株）共分为15个血清型,9个血清群,有8株不能进行K分型,不可分型率为18.2%。食源性疾病分离株分为4个血清型、3个血清群,主要血清群为O3,其次为O1;食品分离株分为11个血清型、6个血清群,主要血清群为O2,其次为O1和O3。44株VP对15种抗生素的药敏结果显示,头孢唑啉耐药率高达68.2%,其次是氨苄西林22.7%、红霉素13.6%、阿奇霉素4.5%、头孢西丁2.3%、头孢噻肟2.3%。菌株主要对β-内酰胺类和大环内酯类抗生素耐药,耐药性逐年增强,一些食源性疾病分离株出现多重耐药,且多重耐药株数量逐年增加。食源性疾病分离株较食品分离株耐药率高,血清型O3∶K6为主要流行菌株,且耐药率高,PFGE分子分型按相似度90%以上,可分为5种PFGE优势带型。食源性疾病分离株相似度较高,可聚类成3个组。而食品分离株相似度较低,除了两株菌相似度大于90%外,其余菌株间相似度均在85%以下。结论 O3与O1血清型菌株之间有高度同源性,提示O3与O1血清型菌株密切相关。食品分离株同源性较低,提示食品株间关联性较差。辽宁省2020年VP分离株耐药情况不容乐观,耐药趋势还需进一步关注。     

江苏省食源性副溶血性弧菌毒力基因和同源性分析研究

目的了解江苏省不同来源的副溶血性弧菌菌株的毒力基因携带情况以及分子遗传特征。方法对江苏省食源性疾病监测病人粪便、疾病暴发和食品监测分离到的副溶血性弧菌进行毒力基因和PFGE分子分型分析。结果暴发病人分离株92.3%携带毒力基因,监测病人分离株96.3%携带毒力基因。暴发食品分离菌株30%携带毒力基因,食品监测分离株不携带毒力基因。101株副溶血性弧菌经PFGE分子分型后,共形成43个带型模式(P1～P43),其中P9带型为最主要的带型模式;按相似度85%为判读标准可分为24个带型簇(cluster),G带型簇为最主要的带型簇(n=55)。结论病人分离株携带毒力基因的比例远高于食品分离株的比例,暴发病人分离株与疾病监测病人分离株携带毒力基因的比例无明显差异。PFGE结果显示我省副溶血性弧菌主要为G带型簇,以P9为主要带型模式,此带型模式为我省副溶血性弧菌的优势带型,在我省存在持续的传播。     

宁波地区食品中致病菌监测与流行株分析

目的了解宁波地区食品中致病菌检出情况和菌株的耐药性,发现其流行优势菌。方法致病菌检测采用直接分离与增菌分离相结合的方法;细菌鉴定采用生化筛检和API等方法;细菌分型采用诊断血清和PFGE基因分型;药敏试验采用K-B法,耐药基因检测采用PCR法。结果 6 812份食品样品中检出目标菌7类12种,共2 331份,检出率为34.22%,致病性弧菌检出数最高,其次为沙门菌和致病性气单胞菌。副溶血性弧菌与其他致病菌差异有统计学意义(P0.01),分离出10个血清群和29个PFGE型,其中O6、O5血清群和PFGE 1型是副溶血性弧菌的主要优势流行型。检出的致病菌对大多数抗生素敏感,其中3株气单胞菌为带aacc耐药基因的多重耐药菌。结论宁波地区食品中致病菌种类较多,易引起食源性疾病;各类致病菌均有流行优势株,副溶血性弧菌是最主要的流行优势株;血清分型和PFGE型能发现优势菌,但均有一定的局限性。     

食品来源副溶血性弧菌毒力基因分布及分子分型研究

分析食品中分离获得的副溶血性弧菌主要毒力基因的分布特征、血清型及分子分型特点,为食源性副溶血性弧菌感染暴发的早期预警和疫情溯源提供实验室数据。提取40株副溶血性弧菌基因组,分别用PCR检测VcrD1、Vp1680、VopP和VcrD2基因,荧光PCR检测tdh、trh和tlh基因。对40株菌株进行血清型分型并采用脉冲场凝胶电泳（PFGE）对其进行分子分型。利用BioNumerics软件对图谱进行聚类分析,建立PFGE分子分型数据库。40株菌株均未检测到tdh、trh、Vop P和VcrD2基因,tlh、VcrD1和Vp1680基因阳性检测率为100%。40株菌株中含有16种血清型,其中9株未分型,主要血清型为O₂∶K₂₈和O₅∶K₁₇,血清型分布呈多样性。40株菌株共获得39个不同的PFGE带型,PFGE呈现遗传多样性,无优势带型。本实验食品环境中分离的副溶血性弧菌未发现毒力因子,副溶血性弧菌血清型呈分散趋势且菌株之间亲缘关系较远。      

北京市人源性单核细胞增生李斯特菌耐药特征及分子分型研究

目的研究北京市人源性单核细胞增生李斯特菌的血清学分型、耐药及分子特征。方法对2013—2015年北京市李斯特菌病专项监测中分离的32株单核细胞增生李斯特菌进行PCR法血清学分型和PFGE分型,并采用CLSI和EUCAST推荐的微量肉汤稀释法检测菌株对12种抗生素的敏感性。结果 32株单核细胞增生李斯特菌共分为4种血清型,其中位居前2位的血清型是1/2b和1/2a,分别为17株和12株,4b血清型2株,1/2c血清型1株。32株菌经Asc I酶切共分为23种PFGE带型,分为4个簇,相同血清型李斯特菌的PFGE带型聚集成簇,两种分型方法一致性为93.75%(30/32)。32株单核细胞增生李斯特菌均对青霉素、氨苄西林、复方新诺明及红霉素敏感,大部分菌株对其他8种抗生素的MIC值较小,但有少数菌株对苯唑西林和四环素的MIC值较大,高达32、64μg/ml。结论北京市人源性单核细胞增生李斯特菌血清型以1/2b和1/2a为主,PFGE图谱与血清学分型存在一定相关性,耐药率普遍较低,但需加强耐药趋势监测。     

2012—2014年无锡市不同来源副溶血性弧菌的病原学特征分析

目的了解近年来无锡市不同来源的副溶血性弧菌的毒力基因携带情况、血清型和耐药性。方法对于分别来源于食源性疾病事件、哨点医院监测标本、外环境监测样品的92株副溶血性弧菌分离株,采用PCR进行毒力基因鉴定,玻片凝集试验进行血清分群,纸片法进行耐药试验。结果 92株副溶血性弧菌中,食源性疾病事件、哨点医院、外环境监测这三种来源的分离株中携带tdh基因的菌株分别占85.7%(12/14)、88.6%(39/44)和14.7%(5/34);食源性疾病事件分离株和哨点医院分离株血清群以O3为主,分别占比85.7%(12/14)和65.9%(29/44),外环境监测分离株无优势血清群;食源性疾病事件、哨点医院、外环境监测这三种来源的分离株中耐氨苄西林的菌株分别占35.7%(5/14)、65.9%(29/44)和11.8%(4/34)。不同来源的菌株对氯霉素、氨苄西林/舒巴坦、庆大霉素、亚胺硫霉素都表现为100.0%敏感。结论 2012—2014年无锡市从食源性疾病事件、哨点医院监测标本和外环境监测样品分离出的副溶血性弧菌在毒力基因、血清群、耐药情况上均有较大差异。建议结合耐药试验结果,在无锡市临床治疗副溶血性弧菌感染病例时,抗生素使用首选氯霉素、氨苄西林/舒巴坦、庆大霉素、亚胺硫霉素。     

食品来源副溶血性弧菌毒力基因分布及分子分型研究

分析食品中分离获得的副溶血性弧菌主要毒力基因的分布特征、血清型及分子分型特点,为食源性副溶血性弧菌感染暴发的早期预警和疫情溯源提供实验室数据。提取40株副溶血性弧菌基因组,分别用PCR检测VcrD1、Vp1680、VopP和VcrD2基因,荧光PCR检测tdh、trh和tlh基因。对40株菌株进行血清型分型并采用脉冲场凝胶电泳(PFGE)对其进行分子分型。利用BioNumerics软件对图谱进行聚类分析,建立PFGE分子分型数据库。40株菌株均未检测到tdh、trh、Vop P和VcrD2基因,tlh、VcrD1和Vp1680基因阳性检测率为100%。40株菌株中含有16种血清型,其中9株未分型,主要血清型为O_2∶K_(28)和O_5∶K_(17),血清型分布呈多样性。40株菌株共获得39个不同的PFGE带型,PFGE呈现遗传多样性,无优势带型。本实验食品环境中分离的副溶血性弧菌未发现毒力因子,副溶血性弧菌血清型呈分散趋势且菌株之间亲缘关系较远。     

一起由副溶血性弧菌引起的聚集性腹泻事件的病原特征分析

目的 对2021年湖州市南浔区一起聚集性腹泻事件分离的副溶血性弧菌进行病原特征分析，为及时采取控制措施提供依据。方法 通过询问患者、医院核查等方法搜索到疑似病例59例，对采集的29份厨师、服务员和病例的肛拭子标本以及18份留存餐食进行病原学检测，采用血清学鉴定、脉冲场凝胶电泳（PFGE）、多重荧光聚合酶链式反应和微量肉汤稀释法分别对副溶血性弧菌分离株进行血清分型、分子分型、tlh/tdh/trh毒力基因检测和药敏试验。结果 从3份厨师和6例病例的肛拭子标本中分离到副溶血性弧菌，检出率为15.25%；分离株血清型均为O10:K4。所有菌株tlh、tdh基因均为阳性，trh基因均为阴性。9株分离株仅对氨苄西林耐药，对其他12种抗生素均敏感。经PFGE聚类分析，菌株间相似系数为100.0%，厨师分离株和病例分离株带型一致。结论 这是一起由O10:K4副溶血性弧菌引起的聚集性腹泻事件，分离株均携带tdh毒力基因，分离株之间的同源性高。     

Real-time PCR von Enterobacter sakazakii in Säuglingsanfangsnahrung

Enterobacter sakazakii represents a rare opportunistic pathogen that causes a high mortality rate. Especially newborns are infected by contaminated infant formula. To reduce this risk, the EU directive 2073/2005 demands for the absence of the pathogen in 30 ×10 g infant formula. We describe a PCR-/real-time PCR that specifically detected 32 E. sakazakii strains among 21 species of Enterobacteriaceae as well as contaminated specimens among 30 different brands of infant formula. This real-time PCR, performed after a selective enrichment of E. sakazakii, reduces the working time by 1 to 3 days in comparison to the detection methods recommended by the US Food and Drug Administration (FDA) and ISO/DTS 22964 that is still under revision. In contrast to the latter procedure, we detected contaminated specimens of infant formula by the use of the procedure described in this report and the FDA method.

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Zusammenfassung: Enterobacter sakazakii repr?sentiert einen seltenen opportunistischen Erreger, der Infektionen mit hoher Mortalit?tsrate hervorruft. Insbesondere Neugeborene erkranken durch kontaminierte Anfangsnahrung. Zur Risikominimierung fordert die EU-Verordnung 2073/2005 die Abwesenheit des Erregers in 30 ×10 g S?uglingsanfangsnahrung. Ein PCR-/Real-Time PCR-Verfahren wird beschrieben, das zun?chst mit 32 E. sakazakii -St?mmen und weiteren 20 Arten der Enterobacteriaceen geprüft und anschlie?end zur Untersuchung 30 unterschiedlicher Marken von S?uglingsanfangsnahrung verwendet wurde. Nach der Selektivanreicherung führt das Real-Time PCR-Verfahren zu einer Ersparnis von 1 bis 3 Arbeitstagen beim Nachweis von E. sakazakii im Vergleich zum Verfahren der amerikanischen Food and Drug Administration (FDA) und dem noch in Bearbeitung befindlichen Verfahren ISO/DTS 22964. Im Gegensatz zur letztgenannten Untersuchungsmethode erfolgten mehrere Positivnachweise mit dem hier beschriebenen Verfahren und dem der FDA.

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Eingegangen: 18. Januar 2007     

Occurrence of foodborne pathogens in Irish farmhouse cheese

Food safety is a critical factor in the production of farmhouse cheese. In Ireland the varieties of farmhouse cheese produced reflect a much broader range than those produced commercially and some of these cheese varieties are associated with greater microbiological risk. These include cheese produced from unpasteurised milk and soft ripened cheese such as mould or smear-ripened cheeses which have high pH and relatively short ripening times. The aim of this study was to determine the microbiological quality of farmhouse cheeses in Ireland. Three hundred and fifty one cheese samples, from 15 cheese producers, were analysed for microbiological quality on a monthly basis throughout the year. The analyses included enumeration of Escherichia coli, Staphylococcus aureus and Listeria monocytogenes (using the relevant agars) and enrichment for L. monocytogenes. The cheeses selected were produced from ovine, caprine and bovine milk. Both unpasteurised and pasteurised milk cheeses were sampled and these included hard, semi-hard and soft cheeses, internal/external mould-ripened and smear-ripened cheeses and the cheeses represented different geographic regions. Of the cheeses tested, 94% were free of L. monocytogenes, all were within the EU limits for E. coli and only one cheese variety had S. aureus levels above the recommended numbers for the first 6 months of the year. Due to a modified production process the numbers were within the guidelines for the second six months. The results indicate that Irish farmhouse cheeses are of a high microbiological quality.     

Formation of cereulide and enterotoxins by Bacillus cereus in fermented African locust beans

Afitin, iru and sonru are three spontaneously fermented African locust bean Benin condiments. The fermentation processes are exothermic, with temperatures mostly being above 40 °C. A total of 19 predominant Bacillus cereus isolates from afitin, iru and sonru, were investigated. The enterotoxin genes nhe (A, B, C) were present in all 19 isolates, the hbl (A, C, D) in one (afitin), and the cytK gene in three isolates (afitin). Levels of cytotoxicity to Vero cells and NheA production in BHI-broth was within the range of known diarrheal outbreak strains. Autoclaved cooked African locust beans inoculated with emetic (cereulide producing) B. cereus Ba18H2/RIF supported growth at 25, 30 and 40 °C with highly different maximum cereulide productions of 6 ± 5, 97 ± 3 and 0.04 ± 0.02 μg/g beans, respectively (48 h). For non-autoclaved cooked beans inoculated with 2, 4 and 6 log₁₀B. cereus Ba18H2/RIF spores/g beans, cereulide production was 5 ± 4, 64 ± 8 and 69 ± 34 μg/g beans, respectively at 24 h, while it was 70 ± 43, 92 ± 53 and 99 ± 31 μg/g at 48 h of fermentation at 30 °C. Even though high toxin levels were observed, to date there are no known reports on diarrhea or vomiting due to the consumption or afitin, iru and sonru in Benin, which also according to the present study is likely to be expected from the low levels of cereulide produced at 40 °C.     

佳木斯市肠炎沙门菌的耐药特征及同源性分析

目的 了解佳木斯市优势血清型肠炎沙门菌的耐药状况及进化关系,为临床治疗提供科学依据。方法 对来自食品和腹泻患者的沙门菌进行血清学分型,采用微量肉汤稀释法对分离的肠炎沙门菌进行16种抗生素药敏试验,脉冲场凝胶电泳分析菌株之间的进化关系。结果 在108株沙门菌中,检出肠炎沙门菌55株（50.93%）;49株肠炎沙门菌有耐药性,耐药率为89.09%,其中33株菌（67.35%）为多重耐药,对抗生素萘啶酸和氨苄西林的耐药率较高,分别为81.82%和60.00%;55株肠炎沙门菌同源性为37%~100%。结论 佳木斯地区肠炎沙门菌多重耐药率高,菌株之间亲缘关系和耐药性存在密切关联性,来自食品和腹泻患者的肠炎沙门菌有交叉感染现象,依据本地区肠炎沙门菌耐药特征,制定有针对性诊疗方案和预防控制措施,对保障公共卫生健康发展具有重要意义。     

Cloning and in vitro and in vivo expression of plant glutathione S-transferase zeta class genes

The cDNAs encoding the zeta class of glutathione S-transferases (GSTs) (GSTZs), which are multifunctional enzymes, were cloned from Arabidopsis thaliana L. and three lines of Brassica napus L. by RT PCR, and named AtGSTZ for A. thaliana ecotype Columbia gll, and BnGSTZ-A, BnGSTZ-B and BnGSTZ-C for B. napus cv. Shan 2A, Shan 213 and Ken C1, respectively. Sequence analysis revealed that the sequences of Shan 2A and Shan 2B were identical, and Ken C1 was different only in 3.8% of the sequence. Comparison of these sequences with published sequences in GeneBank showed that the sequences of the Brassica species were unpublished. The cDNAs were then inserted into two vectors for in vivo expression in Escherichia coli and in vitro expression in a wheat-germ cell-free protein synthesis system. All GSTZs were well expressed both in vivo and in vitro as an enzymatically active form, showing that all of them had both dichloroacetic acid dechlorinating and maleylacetone isomerase activities.     

2022年上海市生禽肉和调理肉制品中主要食源性致病菌监测

目的 分析上海市市售生禽肉和调理肉制品中沙门菌、单核细胞增生李斯特菌及小肠结肠炎耶尔森氏菌等食源性致病菌的污染情况。方法 2022年1～8月,从上海市农贸市场、超市、餐饮店等环节采集样品348份,其中生禽肉240份,调理肉制品108份。按照食品安全国家标准分别对样品中的沙门菌、单核细胞增生李斯特菌和小肠结肠炎耶尔森氏菌的检测。采用VITEK2全自动微生物鉴定仪对可疑菌株进行生化鉴定,并对沙门菌分离株进行血清学分型。结果 生禽肉中沙门菌、单核细胞增生李斯特菌和小肠结肠炎耶尔森氏菌的检出率分别为28.33%(68/240)、5.00%(12/240)和0.83%(2/240);调理肉制品中沙门菌、单核细胞增生李斯特菌和小肠结肠炎耶尔森氏菌的检出率分别为5.56%(6/108)、28.70%(31/108)和1.85%(2/108);血清学分型结果显示,生禽肉中68株沙门菌分布于14种不同血清型,其中科瓦利斯沙门菌（35.29%）、鼠伤寒沙门菌（16.18%）和肠炎沙门菌（13.24%）占比较高,调理肉制品中6株沙门菌分布于3种不同血清型,分别为肠炎沙门菌（66.67%）、肯塔基沙门菌（16...     

Non-nutritive functional agents in rattan-shoots, a food consumed by native people in the Philippines

The tender shoots of Calamus ornatus, one of the food items consumed by the native people, Kanawan Aytas, in the Bataan region of the Philippines, have not been studied before. A bioassay-guided investigation of its methanolic extract afforded non-nutritive functional agents (NFAs), steroidal saponins 1–3, along with its aglycone (4). The NFAs 1–4 inhibited cyclooxygenase enzymes, COX-1 and -2, by 47%, 43%, 33%, and 53% and 71%, 75%, 78%, and 73%, respectively, at 28.2, 24.2, 21.2 and 60.4 μM. Treatment of breast (MCF-7), CNS (SF-268), lung (NCI-H460), colon (HCT-116) and gastric (AGS) cancer cell lines with the extract at 100 μg/ml reduced cell proliferation. Similarly, the pure NFAs 2 and 3 reduced the cell viability of breast, CNS, lung, colon and gastric cancer cell lines by 37.5%, 22.4%, 53.3%, 58.2%, 40.3% and 29.8%, 21.3%, 45.6%, 37.1%, 25.0%, respectively, at 24.2 and 21.2 μM. The 50% reduction in cell viability (IC₅₀) concentrations of 2 and 3 against these cancer cell lines were 8.8, 6.1, 7.5, 23.8, 12.1 and 3.8, 7.1, 3.3, 14.3, 12.1 μM, respectively. This is the first report on the isolation of steroidal saponins from C. ornatus shoots and their antiinflammatory and tumor cell proliferation inhibitory activities. Therefore, our results suggest that the Kanawan Aytas may yield health benefits from rattan-shoots in their diet.     

Assessment of the microbiological safety of salad vegetables and sauces from kebab take-away restaurants in the United Kingdom

The purpose of this study was to establish the microbiological safety of salad vegetables and sauces served in kebab take-away restaurants. Comparison with published microbiological guidelines revealed that 4.7% of 1213 salad vegetable samples were of unsatisfactory microbiological quality due to Escherichia coli and/or Staphylococcus aureus levels at ≥10² cfu g⁻¹. Another 0.3% of salad samples were of unacceptable quality due to S. aureus at ≥10⁴ cfu g⁻¹ (2 samples) or the presence of Salmonella Kentucky (1 sample). Cucumber was the most contaminated salad vegetable with regards to unsatisfactory levels of E. coli (6.0%) or S. aureus (4.5%). Five percent of 1208 sauce samples were of unsatisfactory microbiological quality due to E. coli, S. aureus at ≥10² cfu g⁻¹ and/or Bacillus cereus and other Bacillus spp. at ≥10⁴ cfu g⁻¹. A further 0.6% of sauce samples were of unacceptable quality due to Bacillus spp. (Bacillus subtilis, Bacillus pumilus, Bacillus licheniformis) at ≥10⁵ cfu g⁻¹ or the presence of Salmonella Agbeni (1 sample). More samples of chilli sauce (8.7%) were of unsatisfactory or unacceptable microbiological quality than any other sauce types. The results emphasize the need for good hygiene practices in kebab take-away restaurants handling these types of ready-to-eat products.