多黏菌素类抗生素耐药性编码基因检测用质粒标准样品研制

目的 研制适合多黏菌素类抗生素耐药性编码基因检测用质粒DNA标准样品。方法 由National Center for Biotechnology Information（NCBI）数据库检索得到mcr-1、mcr-3、mcr-5基因参考序列,构建重组质粒和重组菌株;将重组菌传代至15代检测目标基因的遗传稳定性。提取重组质粒,真空干燥,制得标准样品。将标准样品水化、连续10倍梯度稀释,测定聚合酶链式反应（PCR）和实时荧光PCR（RT-qPCR）扩增目标基因的检出限。随机抽取质粒DNA标准样品,采用PCR法和紫外分光光度计法检验样品的均匀性及其在4 ℃、37 ℃、-20 ℃的贮存稳定性。结果 成功获得多黏菌素类抗生素耐药机制编码基因mcr-1、mcr-3、mcr-5基因片段,重组菌15代传代中目的基因遗传稳定。mcr-1、mcr-3、mcr-5标准样品PCR和RT-qPCR最低检出限分别为1.67×10⁴ 拷贝数/μL和1.67 拷贝数/μL、1.31×10⁶ 拷贝数/μL和13.1 拷贝数/μL、1.55×10⁵ 拷贝数/μL和1.55 拷贝数/μL。T检验表明,各基因随机抽取的12管标准样品质量间的F值均小于F_临界值,且PCR均可检出,均匀性符合要求。标准样品在4 ℃贮存90 d、37 ℃贮存14 d、-20 ℃贮存360 d,定性、定量检验结果稳定、无极显著性差异。结论 本实验制备的mcr-1、mcr-3、mcr-5质粒DNA标准样品目的基因遗传稳定,均匀性和贮存稳定性良好。本研究结果可用于多黏菌素类抗生素耐药机制的快速预测和相关基因检测的质控样品。     

食品检测用肠道侵袭性大肠埃希氏菌标准物质的研制

目的 为满足肠道侵袭性大肠埃希氏菌(enteroinvasive Escherichia coli, EIEC)准确检测的实验室质量控制和能力验证需求,研制具有我国自主知识产权、稳定性良好且具有清晰基因组信息背景的即用型EIEC标准物质。方法 利用二代高通量测序技术对EIEC(CMCC 44840)进行全基因组测序,明确CMCC 44840的种属、血清型、多位点序列分型(MLST)和毒力基因;采用冷冻干燥技术制备活菌含量为10³ CFU的EIEC冻干样品;参照CNAS-GL017-2018进行均匀性检验,并采用单因素方差分析对结果进行统计分析;将样品分别于-20 ℃、4 ℃、25 ℃和37 ℃条件下保藏,对其储藏稳定性和运输性进行评价;利用5种食品基质样本进行标准物质的使用效果验证,同时组织3家实验室进行协同标定。结果 CMCC 44840基因组大小为4.96 Mb,GC含量为50.7%,编码基因5 424个,种属鉴定结果为大肠埃希氏菌,(Escherichia coli)血清预测结果为O28ac:H7,MLST为ST311型,携带ipaH、virB、virF等毒力基因;制备的EIEC冻干样品均匀性检验结果F=1.79,符合标准物质均匀性要求;冻干样品在25 ℃和4 ℃下保藏7 d,-20 ℃和4 ℃下保藏60 d,菌含量均保持在10³CFU水平,在37 ℃下第3 d菌含量出现下降,低于10³CFU水平;添加EIEC冻干样品的20件不同食品基质样品经增菌后均检出EIEC;3家协同标定实验室测定EIEC冻干样品菌含量均为10³ CFU,且实验室间无显著性差异(F=0.59)。结论 本研究所制备的EIEC标准物质所用菌株具有清晰的基因组序列信息,均匀性和稳定性均符合要求,适用性良好,能够满足食品检测实验室的质量控制和能力验证的需求。     

食品检验用解脂耶氏酵母菌标准物质制备研究

目的 为满足实验室酵母菌日常检验质量控制需求及实验室间比对,制备解脂耶氏酵母菌检验用标准物质。方法 通过生化、基质辅助激光解吸电离飞行时间质谱及ITS位点测序对研究用菌株进行菌种鉴定确认后,刮取适宜菌苔于保护剂中混匀后冻干,制备10_⁴ CFU/样品浓度的标准物质。参照CNAS-GL003等标准要求进行均匀性检验后,采用单因素方差分析对结果进行评价。将样品放于25 ℃和37 ℃及-20 ℃、4 ℃条件下,分别进行运输稳定性检验及储存稳定性检验。依据国家标准GB 4789.15,将研制的本标准物质加入7类食品基质共20件样品中进行检验,验证真实食品样品中适用性。组织3家实验室,对本标准物质进行协作标定。结果CMCC98025经生化鉴定为解脂耶氏酵母菌,准确度为93%;MALDI-TOF MS鉴定结果为解脂耶氏酵母,分数为2.012;ITS位点测序结果与NCBI Genbank中已有序列比对,匹配最优结果为解脂耶氏酵母Yarrowia lipolytica(Accession number ：CP061015.1;Query Cover：95%;Ident：100%)。均匀性测试结果符合正态分布,通过单因子方差分析计算F_0.05(19,20)=2.137,F值为1.697,F＜F_0.05,符合均匀性要求。25 ℃及37 ℃培养箱中放置7天,标准物质中菌含量仍保持在10_⁴ CFU/样品,可保持稳定。标准物质于4 ℃储存28天及-20 ℃储存90天,菌含量为10_⁴ CFU/样品,复苏率均在91%以上,说明4 ℃放置较短时间及-20 ℃放置较长时间样品稳定。7类食品样品基质中加入本标准物质后进行检验,均可检出,回收率为81.3%。经三家实验室协作标定,本标准物质平均浓度为2.0～3.0×10_⁴ CFU/样品。结论 本研究制备的解脂耶氏酵母菌标准物质均匀性、稳定性、真实食品样品中应用验证及协作标定结果均符合要求,可应用于食品中解脂耶氏酵母菌定性检验,今后可作为实验室日常检验工作的阳性对照质控样品,也可作为实验室间比对样品发放,以保障日常检验工作结果可靠性,进一步提升检验机构人员的检验水平。     

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。     

广西单核细胞增生李斯特菌的流行率、毒力特征和分子分型研究

目的 比较广西不同种类食品中单核细胞增生李斯特菌（以下简称单增李斯特菌）的流行情况、毒力特征和分子分型特征，更好地了解单增李斯特菌的遗传特点及其传播的潜力。方法 对来自2002—2020年的210株单增李斯特菌进行全基因组测序（WGS），分析其序列分型（ST）、克隆群（CC）型及毒力基因。结果 2002—2020年广西53.8%单增李斯特菌分离株来源于肉与肉制品，59.0%分离株来自于谱系Ⅱ，优势ST型为ST8和ST9。79.4%菌株携带LIPI-1基因（hly、prfA、plcA、plcB、mpl、actA），83.7%菌株携带LIPI-2基因（inlC、inlF、inlJ、inlK）。17.6%菌株携带LIPI-3基因（llsA、llsB、llsD、llsG、llsH、llsP、llsX、llsY）。结论 肉与肉制品是广西单增李斯特菌检出率最高的食品类型，分子分型结果证实了单增李斯特菌种群具有高度的遗传多样性，WGS的高分辨力可用于监测食源性单增李斯特菌病的暴发。毒力基因的分布表明广西单增李斯特菌存在不同程度毒力基因的缺失，应警惕高致病性的菌株引起的食源性暴发事件。     

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。     

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。     

可生食蔬菜中单核细胞增生李斯特菌生存特征研究

目的 探究可生食蔬菜品种、温度、接种部位对单核细胞增生李斯特菌（以下简称单增李斯特菌）存活的影响,为可生食蔬菜中单增李斯特菌的风险评估和关键控制措施提供理论依据。方法 以冻干定量单增李斯特菌为菌株来源,以彩椒、洋葱、黄瓜、圣女果和生菜5种可生食蔬菜的表面和切面为单增李斯特菌的接种点,在4 ℃、25 °C条件下培养7 d,定期监测每份样本中的单增李斯特菌的菌量,对其生长情况进行分析。结果 单增李斯特菌冻干菌种不同瓶间菌量均匀（F=1.923,P<0.05）,-20 ℃储存28 d后的复苏率为93.3%±4.2%。在4 ℃条件下,除了彩椒表面、黄瓜切面、生菜表面和生菜切面外,单增李斯特菌在其他蔬菜上放置7 d后均未见显著生长（δ<0.5 log₁₀ CFU/mL）。在25 ℃条件下,单增李斯特菌在彩椒、洋葱、圣女果、生菜以及黄瓜切面上均呈现为支持生长［δ为（1.16±0.35）~（2.68±0.18）log₁₀ CFU/mL］。单增李斯特菌在黄瓜切面、生菜表面和切面放置7 d后,菌量仍持续增长,在生菜的表面和切面生长趋势和浓度基本一致。结论 单增李斯特菌在可生食蔬菜上的存活能力与蔬菜种类、表面与切面、储存温度等条件密切相关,温度的控制对降低其在可生食蔬菜中的风险至关重要。生菜和切后的黄瓜作为单增李斯特菌高风险食品,应引起风险评估的重视。     

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。     

菌株传代及葡萄糖浓度对回复突变实验的影响

目的 探讨菌株传代培养、不同葡萄糖浓度对回复突变实验的影响。方法 实验菌株选择鼠伤寒沙门氏菌TA型菌株, 分别为TA97、TA98、TA100和TA102菌株, 分别使用原代、传代一次、传代二次菌株进行自发回变实验检测。选取TA97菌株分别在标准底层培养基和低糖顶层培养基进行突变的检测。 结果 4种菌原代组、F1组、F2组的自发回变菌落数差异有统计学意义(P<0.05), 且TA97和TA98的F1组、F2组, TA100的F2组不符合国标GB 15193.4-2014《食品安全国家标准 细菌回复突变试验》对回变菌落数的要求; 标准底层培养基与低糖培养基TA97回复突变数有统计学差异(P<0.05), 但均达到国家标准要求。 结论 菌株的传代及葡萄糖浓度对回复突变实验均有影响。     

Degradation of the adsorbent Tenax TA by nitrogen oxides,ozone, hydrogen peroxide,OH radical,and limonene oxidation products

The degradation of the adsorbent Tenax TA was studied qualitatively by sampling oxidants common in indoor air followed by thermal desorption and gas chromatography. A total of 25 degradation products were identified. Several degradation products not reported previously were identified: 9 for nitrogen dioxide; 11 for ozone; 2 for hydrogen peroxide; 12 for hydroxyl radical; 1 for ozone-limonene mixtures, but none for nitrogen oxide. Whereas ozone shows a complex degradation of the adsorbent, hydrogen peroxide and limonene--ozone mixtures show few products. Nitrogen dioxide and the hydroxyl radical behave almost identically and produce 2,6-diphenyl-p-benzoquinone as the major degradation product. Reactant specific degradation products were identified for ozone (11) and nitrogen dioxide (1).     

TA废水处理技术研究

碱减量废水的主要成分是对苯二甲酸(TA)及其钠盐，以生化法和铁碳曝气法对TA废水进行处理研究，得出CODcr=2000mg/L是徽生物降解TA的抑制浓度；pH值为8～9时活性污泥处理TA废水效果最好；铁碳法处理TA废水35h，其CODcr的去除率达到了80％以上。     

Identification and characterization of Clostridium paraputrificum M-21, a chitinolytic, mesophilic and hydrogen-producing bacterium

A strictly anaerobic, mesophilic and chitinolytic bacterial strain, M-21, was isolated from a soil sample collected from Mie University campus and identified as Clostridium paraputrificum based on morphological and physiological characteristics, and 16S rRNA sequence analysis. C. paraputrificum M-21 utilized chitin and N-acetyl- -glucosamine (GlcNAc), a constituent monosaccharide of chitin, to produce a large amount of gas along with acetic acid and propionic acid as major fermentation products. Hydrogen and carbon dioxide accounted for 65% and 35% of the gas evolved, respectively. The conditions for 1 l batch culture of C. paraputrificum, including pH of the medium, incubation temperature and agitation speed, were optimized for hydrogen production with GlcNAc as the carbon source. The bacterium grew rapidly on GlcNAc with a doubling time of around 30 min, and produced hydrogen gas with a yield of 1.9 mol H₂/mol GlcNAc under the following cultivation conditions: initial medium pH of 6.5, incubation temperature of 45°C, agitation speed of 250 rpm, and working volume of 50% of the fermentor. The dry cell weight harvested from this culture was 2.0 g/l.     

Insecticidal effect of spinosad dust, in combination with diatomaceous earth, against two stored-grain beetle species

Laboratory bioassays were carried out to determine the efficacy of spinosad applied alone or combined with the diatomaceous earth (DE) SilicoSec against adult rice weevils, Sitophilus oryzae and confused flour beetles, Tribolium confusum. Efficacy was assessed on wheat and maize at three dosages of spinosad dust formulation (corresponding to 0.0625, 0.1875 and 0.625 ppm of active ingredient [AI] for S. oryzae and to 0.1875, 0.625 and 1.25 ppm of AI for T. confusum), alone or combined with SilicoSec at 150 ppm for S. oryzae and 250 ppm for T. confusum. The mortality of S. oryzae exposed for 14 d on wheat treated with spinosad ranged between 83% and 100%. Conversely, the mortality of S. oryzae on maize treated with DE or on maize treated with lower doses of spinosad dust did not exceed 19% and was only 59% on maize with the highest spinosad dust treatment. Generally, the presence of SilicoSec combined with spinosad did not significantly increase S. oryzae mortality compared with spinosad alone. For T. confusum, mortality on both commodities was lower than for S. oryzae. After 14 d of exposure on wheat, mortality was 14% at the highest dose of spinosad, but increased to 33% in the presence of DE. Similar results were also obtained for T. confusum exposed on treated maize, which indicated a joint action between spinosad and DE. In the case of S. oryzae, the inclusion of DE reduced progeny production in comparison with spinosad alone. Progeny production of T. confusum was relatively low in all treatments, compared to progeny production of S. oryzae. The results of the study show the potential of combination treatments of spinosad dust and DE, but efficacy varies with the target insect species and commodity.     

Nachweis von Clostridium botulinum Typ A, B, E und F mittels real-time-PCR

Clostridium botulinum is a Gram-positive, anaerobic, spore-performing bacterium with the ability to produce under certain conditions a protein with a characteristical neurotoxicity. Intoxications with C. botulinum toxin belong to the rare occuring food-poisonings; the mortality, however, is very high. C. botulinum produce seven different toxin types (type A to G), human intoxications are currently described to caused by toxin type A, B, E and F. C. botulinum is a strictly anaerobic growing bacterium, so the risk for the consumer’s health is mainly due to non-commercially produced food cans. A special form of botulism is the „infant botulism“. In contrast to the botulism of adults, where the disease is caused through toxin-containing food, spores of C. botulinum can sporulate and produce toxin in the intestines of an infant. The source of infant botulism can be honey, because it contains as a natural product C. botulinum spores. Because of the difficult and time-consuming cultural detection of C. botulinum, PCR methods to screen for the toxin genes A, B, E and F, which are relevant in the human medicine, have been used increasingly during the last years. In this presentation two real-time-PCR assays for C. botulinum, which can be applied in the routine laboratory, will be shown.

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Zusammenfassung: Clostridium botulinum z?hlt zu den anaeroben sporenbildenden Bakterien, die unter bestimmten Bedingungen in der Lage sind, sich in Lebensmitteln zu vermehren und ein Protein mit charakteristischer Neurotoxizit?t zu bilden. Intoxikationen mit Clostridium botulinum-Toxin geh?ren zu den seltenen lebensmittelassoziierten Intoxikationen; die Mortalit?t bei einer Erkrankung ist allerdings sehr hoch. C. botulinum produziert sieben unterschiedliche Toxintypen (Typ A-G), wobei für menschliche Erkrankungsf?lle bisher die Toxintypen A, B, E und F beschrieben sind. Da es sich bei den genannten Keimen um strikt anaerob wachsende Bakterien handelt, stellen vor allem nicht kommerziell hergestellte Konserven, wie z. B. Kesselkonserven, ein Risiko für den Verbraucher dar. Als besondere Form des Botulismus wird der so genannte „S?uglingsbotulismus“ beschrieben. Im Gegensatz zur der Erkrankung, die bei Erwachsenen auftritt und die durch die Aufnahme des bereits toxinhaltigen Lebensmittels verursacht wird, k?nnen Sporen von C. botulinum im Darm von S?uglingen auskeimen und dort Toxine bilden. Ursache für den S?uglingsbotulismus ist h?ufig Honig, der als Naturprodukt C. botulinum-Sporen enthalten kann. Da der kulturelle Nachweis von C. botulinum aufwendig und eine endgültige Differenzierung schwierig ist, wird im Bereich der Routinediagnostik seit einigen Jahren verst?rkt mit PCR-Nachweisverfahren gearbeitet, die ein schnelles Screening auf das Vorhandensein der vier in der Humanmedizin relevanten Toxingene A, B, E und F erm?glichen. In dieser Arbeit werden zwei real-time-PCR-Systeme vorgestellt, die im Bereich der Routinediagnostik einsetzbar sind.

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Eingegangen: 19. Januar 2007     

Effect of lactate-enhancement, modified atmosphere packaging, and muscle source on the internal cooked colour of beef steaks

Earlier studies on lactate-mediated colour stability in beef did not address the possible influence on cooked colour. Our objective was to examine the effect of lactate-enhancement, muscle source, and modified atmosphere packaging (MAP) on the internal cooked colour of beef steaks. Longissimus lumborum (LL) and Psoas major (PM) muscles from 16 (n = 16) beef carcasses (USDA Select) were randomly assigned to 4 enhancement treatments (non-injected control, distilled water-enhanced control, 1.25% and 2.5% lactate), and fabricated into 2.54-cm steaks. Steaks were individually packaged in either vacuum (VP), high-oxygen MAP (HIOX; 80% O₂ + 20% CO₂), or carbon monoxide MAP (CO; 0.4% CO + 19.6% CO₂ + 80% N₂), and stored for 0, 5, or 9 days at 1 °C. At the end of storage, surface and internal colour (visual and instrumental) was measured on raw steaks. Steaks were cooked to an internal temperature of 71 °C, and internal cooked colour (visual and instrumental) was evaluated. Lactate-enhancement at 2.5% level resulted in darker (P < 0.05) cooked interiors than other treatments. Interior cooked redness decreased (P < 0.05) during storage for steaks in VP and HIOX, whereas it was stable for steaks in CO. Our findings indicated that the beef industry could utilise a combination of lactate-enhancement and CO MAP to minimise premature browning in whole-muscle beef steaks.     

Characteristics of Rhodococcus pyridinovorans PYJ-1 for the biodegradation of benzene, toluene, m-xylene (BTX), and their mixtures

The degradation rates of benzene, toluene, and m-xylene (BTX) by Rhodococcus pyridinovorans PYJ-1, were highest at 30, 50, and 25 mg/l, respectively. The degradation rate was highest for toluene (0.106 mg/mg-protein x h) followed by benzene and m-xylene at the optimum pH and temperature of 7 and 32 degrees C, respectively. For BTX mixtures, toluene was the preferred substrate, but degradation of each BTX was competitively inhibited by other BTX compounds. The degradation rate of each component of in the BTX mixture decreased by 57-89% depending on the concentration (1-5 mg/l) of the component compared with that of the component as a single substrate.     

Behavior of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Typhimurium in teewurst, a raw spreadable sausage

The fate of Listeria monocytogenes, Salmonella Typhimurium, or Escherichia coli O157:H7 were separately monitored both in and on teewurst, a traditional raw and spreadable sausage of Germanic origin. Multi-strain cocktails of each pathogen (ca. 5.0 log CFU/g) were used to separately inoculate teewurst that was subsequently stored at 1.5, 4, 10, and 21 °C. When inoculated into commercially-prepared batter just prior to stuffing, in general, the higher the storage temperature, the greater the lethality. Depending on the storage temperature, pathogen levels in the batter decreased by 2.3 to 3.4, ca. 3.8, and 2.2 to 3.6 log CFU/g for E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively, during storage for 30 days. When inoculated onto both the top and bottom faces of sliced commercially-prepared finished product, the results for all four temperatures showed a decrease of 0.9 to 1.4, 1.4 to 1.8, and 2.2 to 3.0 log CFU/g for E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively, over the course of 21 days. With the possible exceptions for salt and carbohydrate levels, chemical analyses of teewurst purchased from five commercial manufacturers revealed only subtle differences in proximate composition for this product type. Our data establish that teewurst does not provide a favourable environment for the survival of E. coli O157:H7, S. Typhimurium, or L. monocytogenes inoculated either into or onto the product.     

Biosynthesis of PP-V, a monascorubramine homologue, by Penicillium sp. AZ

The biosynthetic pathway of PP-V, a new monascorubramine homologue, was elucidated by ¹³C-labeling studies. The [1-¹³C] of acetate was incorporated into 2-, 3a-, 4a-, 6-, 8-, 9-, 11-, 13-, 15-, 17-, and 19-Cs of PP-V, and the [2-¹³C], into 3-, 4-, 5-, 8a-, 9a-, 10-, 12-, 14-, 16-, 18-, and 20-Cs. These incorporation patterns coincide with those reported in the biosynthesis of a Monascus azaphilone pigment, monascorubrin.