市售活鸡和腹泻患者中非伤寒沙门菌分子特征和耐药性研究

分析市售活鸡及腹泻患者中非伤寒沙门菌的分子特征的相似性及耐药性,预防沙门菌的感染流行。方法 收集2012年1月—2013年10月从1 054份腹泻患者粪便样品及440份农贸市场的活鸡肛拭子中分离出的非伤寒沙门菌株93株,进行血清分型,对常见的非伤寒沙门菌采用脉冲场凝胶电泳(PFGE)进行分子分型、Bionumerisc v4.0进行聚类分析、纸片扩散法(K-B法)进行药物敏感性试验。结果 1 054份腹泻样品中共检出非伤寒沙门菌88株,分离率为8.35%,分为25种血清型,肠炎沙门菌居多。其中0～2岁的婴幼儿检出率较高,占52.27%(46/88)。PFGE结果:16株肠炎沙门菌分成14个PFGE型、12株斯坦利沙门菌分成11个PFGE型、14株鼠伤寒沙门菌分成14个PFGE型、6株德尔卑沙门菌分成5个PFGE型;对菌株间的相似性进行比较发现两个100%同源的肠炎沙门菌PFGE型,其他菌株的分子特征的相似性不大,鸡源菌株与人源菌株的分子特征相似性不大;对12种抗菌药物的耐药率在50%以下,不同的血清型耐药率各不相同,耐药较严重的为德尔卑沙门菌。440份活体鸡肛拭子分离出的非伤寒沙门菌5株,分离率为1.14%,共分得4种血清型,其中姆班达卡沙门菌病人中未检出。结论 该地区由非伤寒沙门菌引起腹泻的感染率高,尤以婴幼儿多见,患者与鸡未发现有同一克隆株,对常用抗生素有很好的敏感性。     

腹泻患儿粪便中沙门菌的血清型、药敏分析及脉冲场凝胶电泳分子分型研究

目的 研究2019年北京市朝阳区0-10岁腹泻患儿粪便中分离出的沙门菌的血清型、PFGE分子分型研究及耐药特点。方法 对分离自腹泻患儿病例的47株沙门菌进行血清分型,采用微量肉汤稀释法进行27种抗生素的药敏实验;采用PFGE脉冲场凝胶电泳进行指纹图谱分型研究。结果 47株沙门分为9种血清型,优势血清型两种,分别为肠炎沙门菌23株占48.94%,鼠伤寒沙门菌14株占29.79%。47株沙门菌对磺胺异恶唑耐药率(57.45%)最高,其次为氨苄西林(48.94%)和链霉素(48.94%)。23株肠炎沙门菌可分成8个PFGE指纹图谱,15株鼠伤寒沙门菌可分成14个PFGE指纹图谱。结论 北京市朝阳区0-10岁儿童食源性沙门菌血清主要为肠炎沙门菌和鼠伤寒沙门菌,各血清型的耐药性有所不同,PFGE指纹图谱呈多样性。     

江西省食源性沙门菌血清分型及脉冲场凝胶电泳指纹图谱研究

研究江西省食源性沙门菌的血清型,用脉冲场凝胶电泳(PFGE)方法对优势血清型进行分子分型,建立江西省食源性沙门菌PFGE指纹图谱资料库,为食源性疾病溯源提供数据。方法 对136株食源性沙门菌进行血清分型,对其中优势血清型德尔卑沙门菌菌株进行PFGE分型,获得分子分型指纹图谱,运用Bionumerics v6.6软件进行聚类分析。结果 136株沙门菌分离株血清群以B群为主,优势血清型主要是德尔卑沙门菌(49.26%),67株德尔卑沙门菌可分为41个型别的PFGE指纹图谱(相似值100%的为同一PFGE型别),主要集中在3个型别中。结论 江西省食源性沙门菌血清型分布呈多样性,优势血清型是德尔卑沙门菌,生肉制品是其主要的来源,同种血清型别的PFGE指纹图谱无相关性。     

2016年河南省市售生禽肉中沙门菌血清分型和分子分型研究

目的 了解河南省市售生禽肉中沙门菌污染状况,并对分离株进行血清型和分子分型研究,为河南省食源性疾病溯源数据库提供基础数据。方法 沙门菌检测及血清分型、脉冲场凝胶电泳(PFGE)分子分型分析参照国家食品及食源性疾病监测网工作手册。结果 165份生禽肉样品中检出41株沙门菌,分属13个血清型,优势血清型是科瓦利斯沙门菌、肯塔基沙门菌、鼠伤寒沙门菌及达布沙门菌。经Xba I酶切,获得30种带型,每种带型包括1～5株菌株,相似度为47.5%～100%。部分不同血清型沙门菌PFGE型相似。结论 河南省市售生禽肉沙门菌污染严重,沙门菌血清型和基因型呈现多样化,基因型有一定的地区聚集性。     

两起肠炎沙门菌所致食物中毒的病原学研究及溯源分析

目的探讨两起同地区连续发生的肠炎沙门菌食物中毒分离株之间的分子流行病学关系。方法参照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》方法进行病原分离培养,对检出菌株进行表型鉴定;用VITEK-ⅡCompact全自动微生物分析系统检测菌株抗生素敏感性;采用脉冲场凝胶电泳(PFGE)技术对检出菌株进行分子分型和流行病学特征分析。结果共检出23株肠炎沙门菌,其中从25份患者肛拭样本中检出17株,8份从业人员肛拭样本中检出2株,8份留样食物中检出3株,厨具涂抹样本中检出1株。23株肠炎沙门菌血清抗原式均相同(9∶g,m∶-);生物学性状和药敏试验结果基本一致;PFGE图谱带型完全一致(相似度为100%),且与当地散发病例PFGE图谱带型相似。结论综合流行病学调查、病原学检测和分子分型结果,证实这两起食物中毒是由同一来源的肠炎沙门菌污染所引起。     

2017—2019年绍兴市沙门菌病人分离株的耐药性与分子分型分析

目的 了解绍兴市2017—2019年沙门菌病人分离株的分子分型特征及耐药情况。方法 收集245株分离自绍兴市食源性腹泻病例中的沙门菌,进行血清学分型,采用微量肉汤稀释法进行药敏检测,使用脉冲场凝胶电泳(PFGE)对分离株进行分子分型,利用BioNumerics V7.1软件进行聚类分析。结果 245株沙门菌可分为60种血清型,优势血清型为鼠伤寒沙门菌(32.24%)、肠炎沙门菌(10.20%)和伦敦沙门菌(6.94%)。225株分离株对25种抗生素存在不同程度的耐药,其中氨苄西林(77.14%)、四环素(73.88%)和链霉素(66.53%)的耐药率较高,且多重耐药率达76.33%。Xba Ⅰ酶切后的鼠伤寒沙门菌、肠炎沙门菌和伦敦沙门菌分别含48种、11种和12种不同PFGE指纹图谱。结论 绍兴市沙门菌血清型种类繁多,PFGE指纹图谱呈多样性,抗生素耐药呈现较集中且主要为氨苄西林-四环素-链霉素(AMP-TET-STR)。     

2016—2020年辽宁省沙门菌耐药性及分子分型研究

目的 分析辽宁省2016—2020年食源性疾病监测中沙门菌分离株的耐药情况及分子分型特征,为沙门菌引起感染性腹泻的防控、临床抗生素使用提供可靠科学依据。方法 对辽宁省2016—2020年临床腹泻病例分离的90株沙门菌进行血清学鉴定,应用脉冲场凝胶电泳（PFGE）进行分子分型,采用BioNumeries 7.6软件对菌株间的相似度进行聚类分析,采用微量肉汤稀释法进行药物敏感性检测。结果 90株沙门菌分为13种血清型,以肠炎沙门菌和鼠伤寒沙门菌为主。PFGE聚类分析得到54种带型,各带型间相似度为51.4%～100%,每种带型包含1～10株菌,同一血清型菌株的PFGE带型相似度较高,且存在多次聚集现象。药敏结果显示90株沙门菌呈现36种耐药谱,氨苄西林耐药率最高（66.7%,60/90）,其次为萘啶酸（62.2%,56/90）,头孢西丁全部敏感。多重耐药率达48.9%(44/90),其中鼠伤寒沙门菌和肠炎沙门菌多重耐药率分别为87.5%和45.8%。结论 辽宁省沙门菌引起腹泻病例呈散发态势,菌株多重耐药趋势明显,且耐药率较高,耐药谱广泛,应进一步加强分子溯源及耐药性监测。     

温州市食品中沙门菌污染状况及特征分析

目的了解温州市食品中沙门菌的污染状况,分析分离的沙门菌血清型分布、耐药性及脉冲场凝胶电泳(PFGE)分子分型特征。方法依据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》进行菌株分离鉴定及血清学分型,采用微量肉汤稀释法进行药敏试验,PFGE法进行分子分型。结果 6类食品2 039份样品中,37份样品检出沙门菌,检出率为1.8%,其中生禽肉和生畜肉检出率较高,分别为6.9%(20/290)和3.4%(10/290)。37株沙门菌分属16种血清型,居前三位分别为鼠伤寒沙门菌、德尔卑沙门菌和肠炎沙门菌。81.1%(30/37)的菌株对17种抗生素产生不同程度的耐药,呈现24种耐药谱,多重耐药率为56.8%(21/37)。PFGE图谱分为31种PFGE带型,呈多态性。结论沙门菌在温州市食品中存在一定的污染率,耐药情况形式严峻,PFGE图谱的聚集性与沙门菌的血清型有一定的联系,与耐药谱之间的关联性并不明确。     

2012—2018年广州市海珠区不同来源沙门菌分离株血清学分型及生化分析

目的 了解2012—2018年广州市海珠区沙门菌食品污染物监测分离株、食物中毒分离株、从业人员健康体检分离株的血清学分型及生化分析。方法 2012—2018年分离416株沙门菌,包括28株食物中毒菌株、5株食品污染物监测菌株、383株体检人群菌株,对其进行血清学分型、生化鉴定。结果 416株沙门菌鉴定出40个血清型,其中鼠伤寒沙门菌56株(13.5%),肠炎沙门菌46株(11.1%),德尔卑沙门菌41株(9.9%),其他血清型273株(65.6%)。沙门菌以A～F群血清型为主,同时检出多例罕见血清型。结论 广州市海珠区人群中沙门菌的血清型众多且分布广,鼠伤寒沙门菌、肠炎沙门菌、德尔卑沙门菌为海珠区优势沙门菌血清型；开展沙门菌血清型监测,对该菌引发的疾病有重要意义。     

云南省肠炎沙门菌及德尔卑沙门菌脉冲场凝胶电泳分子分型及耐药性研究

目的 了解云南省沙门菌患者中分离最多的肠炎沙门菌及食品中分离最多的德尔卑沙门菌脉冲场凝胶电泳(pulse field gel electrophoresis, PFGE)分子分型及耐药状况。方法 参照中国细菌性传染病分子分型实验室监测网络Pulse Net China的沙门菌PFGE分子分型方法进行分子分型。分析耐药板最低抑菌浓度(minimal inhibitory concentration, MIC)值, 根据美国临床实验室标准化协会(Clinical and Laboratory Standard Institute, CLSI)的相应标准获得S、I、R结果。结果 56株肠炎沙门菌呈11种PFGE带型, 有4种优势带型, 对氨苄舒的耐药率最高, 为57.14%, 对亚胺培南最敏感。27株德尔卑沙门菌呈25种PFGE带型, 对复合磺胺的耐药率最高, 为64.29%, 对亚胺培南最敏感。结论 肠炎沙门菌分子分型有明显的优势带型, 本地区肠炎沙门菌对氨苄舒的耐药率最高。德尔卑沙门菌分子分型呈多样分布, 复方磺胺是本地区德尔脾沙门菌最耐药抗生素。2种沙门菌都对亚胺培南最敏感。     

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.