2016—2021年无锡市副溶血性弧菌毒力基因和耐药性分析及分子分型研究

目的 分析2016—2021年无锡市不同来源副溶血性弧菌的毒力基因、耐药性和分子分型结果。方法 采用多重荧光PCR、微量肉汤稀释法、脉冲场凝胶电泳（PFGE）分别对204株分离自无锡市各类监测样本中的副溶血性弧菌进行tlh/tdh/trh毒力基因检测、耐药试验和分子分型。数据比较采用χ²检验。结果 204株菌tlh基因携带率为100%（204/204），tdh基因携带率为82.35%（168/204），trh基因携带率为2.45%（5/204），食品及环境分离株与病患分离株tdh基因携带率差异具有统计学意义（P<0.001）。菌株对头孢唑啉耐药率最高达96.08%（196/204），对3种及以上抗菌药物的耐药率为2.94%（6/204），食品及环境分离株与病患分离株对氨苄西林、四环素、磺胺甲??唑/甲氧苄啶、环丙沙星耐药率差异具统计学意义（P<0.05）；204株副溶血性弧菌经过聚类分析，分为123个PFGE带型，相似度49.1%~100.0%，按85%的相似度聚类可分为18个带型簇。结论 无锡市副溶血性弧菌病患分离株大部分携带tdh基因；菌株对头孢唑啉耐药率最高；PFGE型别呈多态性，优势带型不明显。     

2021年贵州省食源性疾病主动监测分离沙门菌耐药及分型特征

目的 了解2021年贵州省食源性疾病主动监测分离沙门菌的血清型、耐药和分子分型特征。方法 对全省2021年食源性疾病主动监测腹泻病例中分离的164株沙门菌采用玻片凝集法进行血清学分型，采用微量肉汤稀释法测定菌株对14种抗生素的最小抑菌浓度（MIC）值，采用脉冲场凝胶电泳（PFGE）进行分子分型。结果 164株沙门菌可分为25种血清型，优势血清型为鼠伤寒沙门菌（76，46.34%）、肠炎沙门菌（25，15.24%）和爪哇安纳沙门菌（15，9.15%）。164株沙门菌耐药率为100%，多重耐药率达86.59%；其中对氨苄西林、四环素和萘啶酸耐药率较高，分别为95.12%（156/164）、78.05%（128/164）和63.41%（104/164）。72株鼠伤寒沙门菌PFGE聚类分析后共分为58种指纹图谱，24株肠炎沙门菌有12种指纹图谱，15株爪哇安纳沙门菌有3种指纹图谱。结论 贵州省腹泻患者沙门菌血清型种类较多，多重耐药现象严重，PFGE指纹图谱表现出遗传多样性。应加强对沙门菌的耐药监测，尤其是优势血清型鼠伤寒沙门菌的临床用药。     

预包装熟肉制品生产加工过程沙门菌污染状况及病原学特征研究

目的 了解预包装熟肉制品生产加工过程各环节中沙门菌污染状况,并分析分离株的分子特征及耐药性。方法 按照国家《熟肉制品（预包装）生产加工过程监测工作手册》采样要求,2015—2017年在德州某预包装熟肉制品厂采集环境样品和熟肉样品共460份,依据现行有效的GB 4789.4—2016进行沙门菌分离鉴定;用血清凝集法对沙门菌进行血清分型;用脉冲场凝胶电泳（PFGE）和多位点序列分型（MLST）法对沙门菌进行分子分型分析;采用微量肉汤稀释法对15种抗生素进行耐药性检测。结果 460份样品中沙门菌检出率为5.65%（26/460）,2016年沙门菌的检出率最高（7.65%,14/183）,不同年份沙门菌检出率差异无统计学意义（χ²=2.82,P>0.05）;中间产品中沙门菌的检出率最高,不同样品属性沙门菌检出率差异有统计学意义（χ²=64.16,P<0.05）;仅在生制品加工车间检出沙门菌,不同车间沙门菌检出率差异有统计学意义（χ²=78.08,P<0.05）。26株沙门菌共分为6个血清型,肠炎沙门菌最多,占53.85%（14/26）。26株沙门菌经PFGE分型后获得12种带型,以S4型为主;经MLST分型共获得5种ST型,ST11为优势型别。26株分离株中有22株对不同的抗生素有耐药性,耐药率最高的是氨苄西林,为73.08%（19/26）,多重耐药率（耐3种及以上抗生素）为73.08%。结论 熟肉制品加工过程中沙门菌污染主要集中在加工过程的原辅料和中间产品环节,产品蒸煮后污染状况可被有效控制。     

2019—2022年聊城市食品和病例分离单增李斯特菌基于全基因组测序的分子特征及耐药性分析

目的 基于全基因组测序（WGS）比较分析2019—2022年聊城市单核细胞增生李斯特菌（Lm）食品和病例分离株基因组特征、毒力性、耐药性以及遗传多样性。方法 对聊城市33株市售食品和临床病例中Lm分离株开展抗生素敏感性试验和WGS。利用MGAP对WGS数据进行拼接组装,对组装基因组进行基因预测和功能注释、MLST,制作cg MLST最小生成树图,并与美国国家生物信息中心（NCBI）上获取的18株国内外Lm构建wg-SNP进化树。结果 33株Lm分离株的基因组大小为2.89～3.41 Mb,CG含量为37.81%～37.97%,可分为6个ST型（ST9、ST121、ST8、ST87、ST155、ST101）,分别属于6个克隆复合群（CC9、CC121、CC8、CC87、CC155、CC101）;分离株均携带fosX和mprF耐药基因,此外还携带lplA1、prsA2等其他18个毒力基因,有不同程度的毒力基因缺失情况。2株菌对四环素耐药,1株菌对林可霉素耐药。均携带毒力岛LIPI-1和LIPI-2,未检测到毒力岛LIPI-3和LIPI-4。wg-SNPs、cgMLST和基于单拷贝核心蛋白序列的系统发育树遗传进化分析显示,33株Lm分子分型呈现高度多样性,病例来源菌株与食品分离株亲缘关系密切,食品分离株与国外暴发分离株在进化关系上密切相关。结论 山东省聊城市食品和病例中分离的单增李斯特菌均携带毒力基因,具有一定的潜在致病能力,耐药情况尚不严重。分子型别呈现出多样性,食品来源菌株和病例分离株具有较近的亲缘关系,提示市售食品有食源性感染的潜在风险。     

温州市食品中小肠结肠炎耶尔森氏菌分布及分子分型研究

目的 了解温州市食品中小肠结肠炎耶尔森氏菌的分子分型及分布特征。方法 4 ℃增菌后用选择性培养基对食品中的小肠结肠炎耶尔森氏菌进行分离鉴定,分析阳性菌株的生物型、血清型、毒力基因型、耐药性和脉冲场凝胶电泳（PFGE）分子型别。结果 采集6类食品,共676份样品,其中68份样品检出69株小肠结肠炎耶尔森氏菌,检出率为10.1%（68/676）。调理肉制品检出率最高（20.5%,9/44）,其次为速冻食品（17.2%,11/64）。95.7%（66/69）的分离株为生物1A型,生物血清型以1A/O∶5（29.0%）为主,其次为1A/O∶8（14.5%）;88.4%（61/69）的菌株仅携带ystB基因,检出1株4/O∶3型菌株携带毒力基因ail、ystA、yadA、virF。分离株对14种抗菌药物的敏感率达到94%以上。32株血清已分型菌株,分为29种PFGE带型。结论 温州市食品中存在一定程度的小肠结肠炎耶尔森氏菌污染,且检出致病性小肠结肠炎耶尔森菌,菌株耐药率处于较低水平,分子分型提示菌株呈高度遗传多态性。     

2021年西安市食源性腹泻患者致泻大肠埃希菌耐药性及分子分型

目的 了解2021年西安市食源性腹泻患者致泻大肠埃希菌（DEC）的感染状况、致病型分布、耐药性及分子分型特征。方法 收集西安市5家哨点医院腹泻患者粪便标本,进行DEC分离和PCR毒力基因分型鉴定,采用微量肉汤稀释法和脉冲场凝胶电泳（PFGE）对分离株进行药敏试验和分子分型,结果采用SPSS 19.0软件进行统计学分析。结果 389份粪便标本检出DEC阳性40份,阳性检出率为10.28%（40/389）。共分离到41株DEC,以肠产毒性大肠埃希菌（ETEC）和肠集聚性大肠埃希菌（EAEC）为主,分别占41.46%（17/41）和39.02%（16/41）。ETEC以estIa/estIb基因型为主（70.59%,12/17）,EAEC以astA/pic基因型为主（87.50%,14/16）。40株菌（97.56%,40/41）对至少1种抗生素耐药,对氨苄西林、四环素、头孢噻肟和萘啶酸耐药率均超过50%,多重耐药率为56.10%（23/41）。41株DEC聚类分析得到40种带型,相似度为62.0%~100.0%。结论 2021年西安市食源性腹泻患者中DEC检出率较高,主要致病型为ETEC和EAEC,PFGE带型较为分散,菌株耐药及多重耐药现象比较严重。     

北京市通州区食源性疾病监测病例中副溶血弧菌的病原特征分析

目的 了解通州区2015—2021年食源性疾病监测病例中副溶血弧菌的脉冲场凝胶电泳（PFGE）优势带型、耐药情况和毒力基因携带情况,为副溶血弧菌感染防控和治疗提供参考数据。方法 对2015—2021年通州区定点监测医院门诊腹泻病例的粪便样本进行副溶血弧菌分离培养,对其进行血清分型、毒力基因检测、PFGE聚类分析及药敏试验。结果 2 828份粪便标本中检出副溶血弧菌100株,检出率为3.54%,每年7~9月是检出高峰月份。不同年份副溶血弧菌检出率差异具有统计学意义（χ²=53.94,P<0.001）。100株副溶血弧菌中有一株未携带tlh基因,89.00%的菌株携带致病性毒力基因tdh。副溶血弧菌优势血清型为O3∶K6（66/100）,其次是O4∶K8（9/100）。98株菌副溶血弧菌（2株降解）PFGE分型结果显示副溶血性弧菌有39个PFGE带型,命名为V1-V39,条带相似度在79.6%~100%之间,基因分布呈高度多态性,V22和V25是通州区副溶血弧菌的优势带型。菌株对头孢唑林的耐药率最高（32.00%）,其次是氨苄西林（14.00%）和多黏菌素E（13.00%）,对四环素类、氯霉素类、氨基糖苷类、碳青霉烯类四类药物100%敏感。结论 通州区2015—2021年食源性疾病监测病例中检出的副溶血弧菌主要是O3∶K6型tdh⁺- trh^-菌株,对头孢唑林,氨苄西林和多黏菌素E耐药。PFGE图谱主要流行菌株带型相似度在93.1%以上,存在暴发风险,食品安全相关部门需做好副溶血弧菌监测及暴发预警工作,预防食源性疾病暴发。     

一起空肠弯曲菌引起食源性疾病事件的病原学研究及溯源分析

目的 对一起食源性疾病事件进行病原菌检测，了解病原菌毒力基因携带情况并进行溯源分析。方法 对事件采集的样本经FilmArray多重PCR系统进行快速初筛，同时进行细菌分离培养鉴定。使用PCR检测技术对分离菌株进行毒力基因检测，采用16S rRNA基因序列分析与PFGE分型方法对分离菌株进行同源性分析。结果 2份患者肛拭子样本和4份食堂厨工肛拭子样本检出空肠弯曲菌，检出菌株均携带flaA、cadF、imaA、cdtA、cdtB、cdtC等毒力基因。16S rRNA基因序列分析表明6株分离菌株均为空肠弯曲菌，1株菌株与其他5株菌株分子发育距离稍远。6株菌株经PFGE分型可分为3种带型，3株菌和2株菌分别呈现同一带型，2种带型相似性为52.2%；另1株菌为另一带型，与其他菌株带型相似性仅为26.7%。结论 实验室结果表明这是一起由不同克隆株的空肠弯曲菌感染引起的食源性疾病事件。     

鸡源大肠埃希氏菌mcr-1基因携带及分析

目的 了解在农业农村部禁止使用多黏菌素作为动物促生长使用后四川部分地区鸡源大肠埃希氏菌（E.coli）mcr-1基因的携带情况,为制定进一步防控措施提供依据。方法 采集四川部分地区市场售卖点肉鸡直肠拭子,用含有多黏菌素（终浓度4 μg/mL）的EC肉汤增菌接种含多黏菌素（终浓度4 μg/mL）的麦康凯平板,挑取可疑菌落,采用PCR方法鉴定菌株并检测mcr-1基因;微量肉汤稀释法测定mcr-1基因阳性菌株对临床常见抗菌药物耐药情况。脉冲场凝胶电泳（PFGE）对mcr-1基因阳性菌株进行同源分析。耐药基因质粒结合实验验证mcr-1基因传播途径。结果 从70份肉鸡样本中的13份检出mcr-1基因阳性大肠埃希氏菌,检出率18.57%（13/70）,对实验的13种抗生素,除13株mcr-1阳性菌株对头孢西丁有12株敏感以外,对其他抗生素都表现出不同程度的耐药,其中四环素和甲氧苄啶/磺胺甲恶唑耐药率最高,达到了100%（13/13）;其次是氨苄西林和氯霉素,耐药率为84.62%（11/13）。PFGE显示13株mcr-1阳性大肠埃希氏菌分属13个不同的型别;质粒结合实验显示mcr-1基因能够通过质粒传播。结论 mcr-1基因在鸡大肠内大肠杆菌中检测率比较高,且鸡大肠中mcr-1阳性大肠埃希氏菌的耐药情况比较严重。     

咸阳市市售食品中单核细胞增生李斯特菌基因组特征及耐药和致病基因分析

目的 采用全基因组测序技术对从咸阳市市售食品中分离的64株单核细胞增生李斯特菌（单增李斯特菌）的基因组特征,耐药和致病性基因进行分析。方法 收集咸阳市市售食品中分离的64株单增李斯特菌,利用微量肉汤法进行药敏测定,同时进行全基因组测序,原始序列经拼接后利用生物信息学软件进行基因组注释、系统发育树构建及基因组特征和遗传原件分析。结果 64株分离株对于氨苄西林、青霉素、美罗培南、复方新诺明、万古霉素5种抗生素结果均为敏感。其中2株分离株对2种抗生素产生抗性,分别为四环素和红霉素。全基因组测序分析表明,64株分离株分属3个谱系,分为15个克隆群（CC）,以谱系Ⅰ和谱系Ⅱ为主;2株耐药株基因型与表型一致,耐药基因上下游遗传环境分析表明,这些基因的可能来源为猪丹毒杆菌、肠球菌和外源质粒。所有分离株均携带致病基因岛LIPI-1和LIPI-2,部分谱系Ⅰ菌株携带LIPI-3或LIPI-4,具有潜在致病风险。携带质粒和抗性相关基因的主要为谱系Ⅱ菌株。inlA基因提前终止突变均发生在谱系Ⅱ,可能降低菌株毒力。结论 咸阳市市售食品中单增李斯特菌基因组结构相对稳定,菌株存在获得性耐药,且因携带更多毒力基因而产生潜在的高致病性菌株。谱系Ⅰ和谱系Ⅱ菌株在毒力基因、抗性相关基因和质粒携带方面均具有差异,显示不同CC型菌株的毒力和环境适应性存在差异,为咸阳市单增李斯特菌的监测和防控提供了参考数据。     

Frequency of DLK1 c.639C>T polymorphism and the analysis of MEG3/DLK1/PEG11 cluster expression in muscle of swine raised in Poland

DLK1 — (Drosophila like element 1) is a paternally expressed gene, associated with the callipyge phenotype in sheep. In a present study we designed a new real-time PCR alleleic discrimination assay for genotyping of a silent C/T mutation (c.639C>T) in DLK1 gene in swine. The DLK1 c.639C>T mutation was highly polymorphic in all breeds analyzed and C allele was predominant in Landrace and Duroc while T allele was more frequent in Pietrain and Pu?awska breed. Moreover, we analyzed mRNA expression of DLK1 and adjacent genes — MEG3 and PEG11 in muscles of swines of different breeds raised in Poland. We did not observe significantly different expression of DLK1, MEG3 or PEG11 mRNA in any of analyzed breeds. We also attempted to assess the effect of DLK1 (c.639C>T) on the expression of genes in callipyge locus but did not find significant differences between animals with alternate genotypes (C/C and T/T homozygotes).     

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.     

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.     

Arsenic content of some edible mushroom species

The arsenic contents of 162 fruit body samples of 37 common edible mushroom taxa were analyzed. The samples were gathered from different habitats of Hungary (mainly from mountains) between 1984 and 1999. The arsenic content of the samples was measured by the inductively coupled plasma spectrometry method. Very low [lower than 0.05 mg/kg dry matter (DM)] concentrations were found in the samples of 13 taxa, while higher (or very high) contents were quantified in other common taxa (the highest arsenic content was recorded in the fruit body of Laccaria amethysthea at 146.9 mg/kg DM). The species of eight genera (Agaricus, Calvatia, Collybia, Laccaria, Langermannia, Lepista, Lycoperdon, Macrolepiota) belong to the so-called accumulating taxa, and this tendency is evident on all habitats. This arsenic accumulation capability is found in two orders of Basidiomycetes (Agaricales and Gasteromycetales), which is to say this phenomenon occurs in the families Agaricaceae, Tricholomataceae and Gasteromycetaceae. The accumulating taxa found all have a saprotrophic type of nutrition; arsenic accumulation is not detectable in xilophagous or in mycorrhizal species. The consumption of the accumulating species found has only a low toxicological risk for three reasons: the consumed fresh fruit bodies contain about a tenfold lower arsenic level than the dried ones, the majority of arsenic occurs not in poisonous inorganic, but in less dangerous (or not poisonous) organic forms, and the frequency of consumption is low.     

Comparison of the most odour-active volatiles in different hop varieties by application of a comparative aroma extract dilution analysis

Application of a comparative aroma extract dilution analysis on the volatiles isolated from five different hops (Hallertau Perle, Hallertau Hersbrucker Spät, Slowenian Golding, Hallertau Smaragd, US Cascade) revealed linalool and myrcene with the highest Flavour Dilution (FD)-factors in all varieties, followed by 2-isopropyl-3-methoxypyrazine, 3-methylbutanoic acid and geraniol. Some odourants, however, showed high FD-factors only in certain varieties, for example, (5Z)-octa-1,5-dien-3-one and germacrene B in Hersbrucker Spät, (3E,5Z)-undeca-1,3,5-triene in Hersbrucker Spät and Cascade and nonanal in Cascade. The overall odour profile of the Cascade sample clearly differed from the other varietes, and was dominated by a black currant like odour note. The identification experiments revealed 4-methyl-4-sulfanylpentan-2-one, so far unknown as hop constituent, as key contributor to this odour. In addition, an odour-active undecatetraene was present, in particular, in Perle and Cascade. Synthesis and structural assignment of the four stereoisomers of (3E)-undeca-1,3,5,9-tetraene allowed the identification of the fresh, pineapple-like smelling compound as (3E,5Z,9E)-undeca-1,3,5,9-tetraene. Among the four isomers synthesised, this compound showed by far the lowest odour threshold of 0.01 ng/L in air.     

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.     

Thermal inactivation of Bacillus cereus and Clostridium perfringens vegetative cells and spores in pork luncheon roll

The aim of this study was to design a thermal treatment(s) for pork luncheon roll, which would destroy Bacillus cereus and Clostridium perfringens vegetative cells and spores. B. cereus and C. perfringens vegetative and spore cocktails were used to inoculate luncheon meat. Samples were subjected to different temperatures and removal times. The decimal-reduction times (D-values) were calculated by linear regression analysis (D = -1/slope of a plot of log surviving cells versus time). The log(10) of the resulting D-values were plotted against their corresponding temperatures to calculate (-1/slope of the curve) the thermal resistance (z-values) of each cocktail. The D-values for vegetative cells ranged from 1 min (60 degrees C) to 33.2 min (50 degrees C) for B. cereus and from 0.9 min (65 degrees C) to 16.3 min (55 degrees C) for C. perfringens. The D-values for B. cereus spores ranged from 2.0 min (95 degrees C) to 32.1 min (85 degrees C) and from 2.2 min (100 degrees C) to 34.2 min (90 degrees C) for C. perfringens. The z-values were calculated to be 6.6 and 8.5 degrees C for B. cereus vegetative and spores, respectively, and 7.8 and 8.4 degrees C for C. perfringens vegetative cells and spores, respectively. The D-values of B. cereus and C. perfringens suggest that a mild cook of 70 degrees C for 12s and 1.3 min would achieve a 6 log reduction of B. cereus and C. perfringens vegetative cells, respectively. The equivalent reduction of B. cereus and C. perfringens spores would require the pork luncheon meat to be heated for 36 s at 105 and 110 degrees C, respectively. The results of this study provide the thermal inactivation data necessary to design a cooking protocol for pork luncheon roll that would inactivate B. cereus and C. perfringens vegetative cells and spores. The data may also be used in future risk assessment studies.     

Diversity of lactic acid bacteria in suan-tsai and fu-tsai, traditional fermented mustard products of Taiwan

Fu-tsai and suan-tsai are spontaneously fermented mustard products traditionally prepared by the Hakka tribe of Taiwan. We chose 5 different processing stages of these products for analysis of the microbial community of lactic acid bacteria (LAB) by 16S rRNA gene sequencing. From 500 LAB isolates we identified 119 representative strains belonging to 5 genera and 18 species, including Enterococcus (1 species), Lactobacillus (11 species), Leuconostoc (3 species), Pediococcus (1 species), and Weissella (2 species). The LAB composition of mustard fermented for 3 days, known as the Mu sample, was the most diverse, with 11 different LAB species being isolated. We used sequence analysis of the 16S rRNA gene to identify the LAB strains and analysis of the dnaA, pheS, and rpoA genes to identify 13 LAB strains for which identification by 16S rRNA gene sequences was not possible. These 13 strains were found to belong to 5 validated known species: Lactobacillus farciminis, Leuconostoc mesenteroides, Leuconostoc pseudomesenteroides, Weissella cibaria, and Weissella paramesenteroides, and 5 possibly novel Lactobacillus species. These results revealed that there is a high level of diversity in LAB at the different stages of fermentation in the production of suan-tsai and fu-tsai.