辽宁省2018年食源性金黄色葡萄球菌脉冲场凝胶电泳分子分型及毒力基因的研究

目的 了解辽宁省食源性金黄色葡萄球菌的肠毒素分布情况及基因分型特征。方法 采用脉冲场凝胶电泳(pulsed field gel electrophoresis, PFGE)和肠毒素分型方法在不同角度和层次对2018年辽宁省内分离出的18株食源性金黄色葡萄球菌进行肠毒素检测与PFGE同源性分析。结果 PCR方法及酶联免疫吸附测定(enzyme-linked immunosorbent assay, ELISA)方法对肠毒素结果显示, 18株菌分别存在SEB、SEC、SED、SEH 4种肠毒素, PFGE聚类分析显示, 18株金黄色葡萄球菌相似系数在60.3%~100%之间。结论 金黄色葡萄球菌均可以用PFGE和PCR进行分型, 都具有较好地分型能力, 辨识度高, 对菌株有很好的溯源性。     

金黄色葡萄球菌分型方法的建立及应用

建立食源性金黄色葡萄球菌的分型与溯源方法。方法 以血浆凝固酶为目标基因,通过PCR方法对食源性金黄色葡萄球菌进行鉴定,同时借助全自动微生物基因指纹鉴定系统(RiboPrinter)对其进行分型和类聚状况分析。结果 PCR方法可特异性鉴定金黄色葡萄球菌,RiboPrinter方法可将金黄色葡萄球菌分为6个亚型,各亚型类聚状况平均分布。结论 上述方法的建立可准确地将金黄色葡萄球菌进行鉴定和分型,分析类聚分布与同源性关系,为食源性致病菌的快速鉴定与分型溯源提供技术手段。     

金黄色葡萄球菌食品分离株肠毒素基因分布及分型研究

目的 对2006-2009年上海市Staphylococcus aureus(S.aureus)食品分离株进行肠毒素基因检测和基因分型研究,以了解肠毒素基因的分布规律及S.aureus的流行特点.方法 利用PCR方法检测食品中金黄色葡萄球菌肠毒素基因,包括5种传统肠毒素基因(SEA-SEE)和4种新型肠毒素基因(SEG-SEJ);利用脉冲场凝胶电泳法对49株食品分离株进行基因分型.结果 本研究发现49株食品分离株中有19株含有肠毒素基因,16株含有传统肠毒素SEA和SEC,且SEC占93.8％,并检测到新型肠毒素SEG、SEI、SEJ和SEH.PFGE法基因分型显示5株菌不能被分型,其余44株可分为28个基因型,表现为基因型的多样性,且分离自不同时间的菌株具有相同的带型.结论 应加强食品中S.aureus的监测分析,为其引起的食物中毒的预防和控制提供科学依据.     

2005—2012年宁波市食源性金黄色葡萄球菌肠毒素基因和多位点序列分型研究

目的了解宁波市食源性金黄色葡萄球菌(S.aureus)肠毒素基因分布和分子分型特征。方法收集2005—2012年宁波市食品中金黄色葡萄球菌菌株,利用聚合酶链式反应(PCR)检测肠毒素A、B、C和D基因(sea,seb,sec和sed),对肠毒素基因阳性菌株利用多位点序列分型(MLST)进行分子分型。结果 2005—2012年共分离菌株190株,肠毒素基因阳性菌株为41株,阳性率为21.58%。4种肠毒素基因阳性率分别为7.37%(14/190)、5.26%(10/190)、8.95%(17/190)和5.79%(11/190),其中13株菌株具有两个及两个以上肠毒素基因。41株菌株可分为12个序列型(ST),以ST5、ST6、ST188和ST1为主,共占75.61%(31/41),在进化树上主要形成4个分支。结论 2005—2012年宁波市食品中金黄色葡萄球菌携带肠毒素基因频率较高,需加强监测。肠毒素基因阳性菌株与中国其他地区食品株在分子分型上存在较大差异。     

广州市白云口岸航空食品中金黄色葡萄球菌凝固酶基因分型及肠毒素基因研究

目的对2013—2015年从广州市白云口岸航空食品中分离的金黄色葡萄球菌进行基因分型研究,为食源性金黄色葡萄球菌分子溯源提供基础数据。方法以血浆凝固酶和肠毒素为目标基因,采用聚合酶链式反应(PCR)方法对9株金黄色葡萄球菌进行基因分型,其中6株为航空食品分离株,1株为配餐车间大门手拭分离株,2株为标准菌株。肠毒素基因检测包括5种传统肠毒素基因(sea、seb、sec、sed、see)和6种新型肠毒素基因(ser、seg、seh、sei、sej、sep)。结果 6株航空食品分离株的血浆凝固酶基因扩增分型结果为2个PCR型,酶切后得3种亚型;肠毒素基因检测结果显示有2株航空食品分离株含有肠毒素基因,检出率为33.3%(2/6),检出的基因为2种传统肠毒素基因(sec、sed)和4种新型肠毒素基因(ser、seg、sei、sej),均同时携带2种以上肠毒素基因。结论血浆凝固酶基因扩增分型结果显示,不同时间、不同采集地点存在相同的基因型,提示金黄色葡萄球菌存在交叉污染的可能性;航空食品分离株共检出6种肠毒素基因,提示金黄色葡萄球菌基因型多样性,应加强其他新型肠毒素基因检测。     

近红外光谱建模法快速测定桔子精油中柠檬烯的含量

为了解日常检测分离株产肠毒素和耐药情况,探讨食源性金黄色葡萄球菌产毒素的类型及分布状况,研究其耐药特性,对本实验室2016年～2017年日常食品检测中的分离株,分别用全自动荧光酶联免疫法检测肠毒素总量和酶联免疫吸附法对肠毒素SEA～SEE进行分型,并用全自动微生物鉴定药敏分析系统进行药敏试验。结果表明,370份食品中分离到的29株金黄色葡萄球菌,产肠毒素的有16株,阳性率为55.2%,其中食物中毒分离株5株都为肠毒素阳性。产2种及以上肠毒素的菌株为12株,占41.4%。A型～E型常见肠毒素都有检出,其中产SEE的菌株最多,有12株,占41.4%,产SEA的菌株次之,为11株。29株食源性金黄色葡萄球菌对庆大霉素、妥布霉素、青霉素、安苄西林、苯唑西林、阿莫西林-克拉维酸、复方新诺名、克林霉素、红霉素、利福平、四环素均有不同程度的耐药,并出现多重耐药性,其中对青霉素和安苄西林的耐药率最高,均为82.8%,其次为红霉素44.8%。该研究的食品监测中分离到的金黄色葡萄球菌其产肠毒素率较高,主要的类型为SEE和SEA。而且分离得到的食源性金黄色葡萄球菌存在不同程度的耐药性和多重耐药现象,建议从各个环节加强监测,降低因耐药菌带来的食品安全风险。     

葡萄球菌肠毒素基因分型PCR检测技术的研究

通过生物信息学分析,设计了不同基因型肠毒素检测引物;应用含溶菌酶的碱裂解法提取葡萄球菌DNA及梯度PCR方法提高了肠毒素检出率和特异性.从国内28株葡萄球菌分离株中共检测出14种不同型的肠毒素基因(sea-see、tsst-1、seg-sel、sek-sel、sen-seo、seq-ser和seu),未检测到sei和sem基因.毒素基因携带率为16.67%,同时携带4种及以上毒素基因的菌株有11株,占39.29%,其中sek、seq和sea毒素基因在所研究菌株中分布最广,分别占到15.50%、14.30%和10.70%.结果表明不同葡萄球菌菌株间毒素型的关联度不同,sek与seq,seb和sed,sec、sei、sen和seu基因型呈现密切相关,各型肠毒素的分布还与菌株分离来源有着密切联系.所建立的PCR方法可用于金黄色葡萄球菌肠毒素基因分型和分布的研究.     

食品基质中不同金黄色葡萄球菌肠毒素基因型表达差异研究

目的 比较研究食品基质中不同金黄色葡萄球菌肠毒素(Staphylococcal enterotoxins ,SEs)基因型的蛋白表达差异,为预防金黄色葡萄球菌食物中毒提供参考依据。方法 采用特异聚合酶链反应方法(polymerase chain reaction,PCR)对食品中分离的金黄色葡萄球菌菌株进行肠毒素基因型检测;选择sea、seb、sec、sed等基因型阳性菌株,分别接种于胰酪大豆胨液体培养基(trypticase soy broth,TSB)、牛奶、鸡肉中,按照国家标准GB 4789.10—2016酶联免疫吸附试验法(enzyme linked immunosorbent assay,ELISA)定量检测TSB培养基、牛奶和鲜鸡肉中不同金黄色葡萄球菌肠毒素基因型的蛋白表达量。结果 30株金黄色葡萄球菌中检测到14株肠毒素基因型阳性菌株,所占比例为46.67%,其中sea基因携带率最高(16.67%),而seb、sec、sed、seh则各占6.67%。SEA 在TSB、牛奶、鸡肉3种基质中的平均表达量为7.37 ng/mL,高于SEB、SEC、SED;不同基质环境对肠毒素的表达具有一定影响,如SEA、SEB、SEC、SED在TSB中的表达水平最高,平均表达量为9.04 ng/mL,牛奶次之,鸡肉最低。结论 肠毒素基因型的表达与菌株自身的调控及环境作用密切相关,本研究对肠毒素的产生机制进行初步了解,有助于进一步降低食物中毒风险。     

鸡肉生产中分离金黄色葡萄球菌的基因分型与耐药性分析

为探究鸡肉制品在加工过程中金黄色葡萄球菌的肠毒素基因分布、分子分型和耐药性情况,本研究对某鸡肉制品加工厂的生产加工过程进行取样,通过DNA提取、PCR扩增nuc基因对金黄色葡萄球菌进行分离鉴定,然后分析肠毒素基因分布情况,对获得带有毒素基因的菌株进行多位点序列分型（MLST）和耐药性研究。结果表明,260份样本中分离到38株金黄色葡萄球菌（其中携带肠毒素基因的菌株有24株）,检出率为63.16%;共检测到7种毒素基因型,其中sed携带率最高（60.53%）,依次为seg（26.32%）、see（21.05%）、sei（18.42%）、sec（10.53%）、sea（7.89%）、seh（5.26%）,携带两种及以上肠毒素基因的菌株有14株（36.84%）。24株携带毒素基因的菌株MLST分型都为ST型,分为3种ST分型,16株为ST7序列型（66.67%）;5株为ST5序列型（20.83%）;3株为ST464序列型（12.5%）。耐药性分析实验结果表明,抑菌效果最好的抗生素是万古霉素,38株菌株都对其敏感（100%）;绝大多数菌株都对抗生素有耐药性,依次为青霉素G（86.84%）、环丙沙星（60.53%）、克林霉素（55.26%）、四环素（52.63%）;对3-6种抗生素耐药的金黄色葡萄球菌菌株分别占18.42%、15.79%、23.68%、7.89%。研究结果表明鸡肉产品中存在金黄色葡萄球菌污染的情况,并且其中存在携带多种类型肠毒素基因的菌株,也存在具有多重耐药性的菌株。     

食物中毒事件中致病菌肠毒素检测和分子分型研究

目的 对一起食物中毒事件样本进行检测,查明食物中毒原因,并对分离菌株进行相关性分析。方法 参照国家标准方法GB4789.10-2016,对采集样本进行细菌学检验。应用酶联免疫吸附法(ELISA)检测葡萄球菌肠毒素。通过脉冲场凝胶电泳(PFGE)进行分子分型,分析菌株之间的相关性。结果 食物中毒事件中5名患者洗胃液、食物、分离菌株,均检测出E型葡萄球菌肠毒素。3株分离菌株PFGE分子分型提示来源不同克隆株,除了产生E型葡萄球菌肠毒素外,还有B、C、D型。结论 该起食物中毒由不同PFGE型别的产毒金黄色葡萄球菌污染食物引起,菌株产葡萄球菌肠毒素的型别不完全相同。金黄色葡萄球菌引起的食物中毒应关注多污染源及菌株肠毒素基因表达情况。     

Reliable identification of grapevine rootstock varieties using RAPD PCR on woody samples

Since grapevine ( Vitis spp .) rootstock material is being traded increasingly as disbudded woody material a lack of distinctive morphological features on such material necessitates an alternative and reliable means of identification. Methods described here were developed for rapid and efficient extraction of DNA from woody samples rich in phenolic compounds and polysaccharides, and for subsequent identification of varieties by RAPD PCR. Using these methods, and with the application of only one selected RAPD primer, we were able to differentiate sixteen rootstock varieties, including the seven varieties most commonly used in Germany. Problems commonly encountered with reproducibility of RAPD patterns were avoided by choosing primers with a dinucleotide sequence and a high G/C content that allowed a rather high annealing temperature of 45°C. Methods described here should also be useful for other horticultural crops, especially those with woody tissues rich in phenolic compounds and polysaccharides.     

Aroma characterization of various apricot varieties using headspace–solid phase microextraction combined with gas chromatography–mass spectrometry and gas chromatography–olfactometry

The characterization of the aromatic profile of several apricot cultivars with molecular tracers in order to obtain objective data concerning the aromatic quality of this fruit was undertaken using headspace–solid phase microextraction (HS–SPME). Six apricot cultivars were selected according to their organoleptic characteristics: Iranien, Orangered, Goldrich, Hargrand, Rouge du Roussillon and A4025. The aromatic intensity of these varieties measured by HS–SPME–Olfactometry were defined and classified according to the presence and the intensity of grassy, fruity and apricot like notes. In the six varieties, 23 common volatile compounds were identified by HS–SPME–GC–MS. Finally, 10 compounds, ethyl acetate, hexyl acetate, limonene, β-cyclocitral, γ-decalactone, 6-methyl-5-hepten-2-one, linalool, β-ionone, menthone and (E)-hexen-2-al were recognized by HS–SPME–GC–O as responsible of the aromatic notes involved in apricot aroma and considered as molecular tracers of apricot aromatic quality which could be utilized to discriminate apricot varieties.     

Tests of potential functional barriers for laminated multilayer food packages. Part I: Low molecular weight permeants

The advent of the functional barrier concept in food packaging has brought with it a requirement for fast tests of permeation through potential barrier materials. In such tests it would be convenient for both foodstuffs and materials below the functional barrier (sub-barrier materials) to be represented by standard simulants. By means of inverse gas chromatography, liquid paraffin spiked with appropriate permeants was considered as a potential simulant of sub-barrier materials based on polypropylene (PP) or similar polyolefins. Experiments were performed to characterize the kinetics of the permeation of low molecular weight model permeants (octene, toluene and isopropanol) from liquid paraffin, through a surrogate potential functional barrier (25 μm-thick oriented PP) into the food simulants olive oil and 3% (w/v) acetic acid. These permeation results were interpreted in terms of three permeation kinetic models regarding the solubility of a particular model permeant in the post-barrier medium (i.e. the food simulant). The results obtained justify the development and evaluation of liquid sub-barrier simulants that would allow flexible yet rigorous testing of new laminated multilayer packaging materials.     

Analysis of benzo[a]pyrene in spiked fatty foods by second derivative synchronous spectrofluorimetry after microwave-assisted treatment of samples

A simple, rapid and inexpensive method has been developed for the determination of benzo[a     

The development of reference materials for paralytic shellfish poisoning toxins in lyophilized mussel. II: Certification study

This paper describes the second part of a project undertaken to develop certified mussel reference materials for paralytic shellfish poisoning toxins. In the first part two interlaboratory studies were undertaken to investigate the performance of the analytical methodology for several PSP toxins, in particular saxitoxin and decarbamoyl-saxitoxin in lyophilized mussels, and to set criteria for the acceptance of results to be applied during the certification exercise. Fifteen laboratories participated in this certification study and were asked to measure saxitoxin and decarbamoyl-saxitoxin in rehydrated lyophilized mussel material and in a saxitoxin-enriched mussel material. The participants were allowed to use a method of their choice but with an extraction procedure to be strictly followed. The study included extra experiments to verify the detection limits for both saxitoxin and decarbamoyl-saxitoxin. Most participants (13 of 15) were able to meet all the criteria set for the certification study. Results for saxitoxin.2HCl yielded a certified mass fraction of <0.07 mg/kg in the rehydrated lyophilized mussels. Results obtained for decarbamoyl-saxitoxin.2HCl yielded a certified mass fraction of 1.59+/-0.20 mg/kg. The results for saxitoxin.2HCl in enriched blank mussel yielded a certified mass fraction of 0.48 +/- 0.06 mg/kg. These certified reference materials for paralytic shellfish poisoning toxins in lyophilized mussel material are the first available for laboratories to test their method for accuracy and performance.     

Announcing a new international peer-reviewed journal:Food Science and Human Wellness now accepting manuscript submission in English

<正>We are pleased to announce the launch of a new international peer-reviewed journal-Food Science and Human Wellness,ISSN 2213-4530,which is an open access journal,produced and hosted by Elsevier B.V.on behalf of Beijing Academy of Food Sciences.Food Science and Human Wellness is an international peer-reviewed English journal that provides a forum for the dissemination of the     

Chinese CTP Market

According to Printing and Printing Equipment Industries Association of China（PEIAC）＇s statistics to the plate manufucturer in China, in 2013, the actual offset plate production has reached 346 million square meters in China. Among them, the CTP production volume was 245 million square meters, up by 11% than that of last year; the total sales of the CTP plate was 239 million square meters, up by 13%.     

Preparatory Work of Print China 2015 is Going Smoothly

正Held at Guangdong Modern International Exhibition Center,Print China 2015 will cover 7exhibition halls,besides the original Hall No.3,4,5,6,7,the newly built F zone of Hall 3 will be used too.The total area will be140,000 square meters.Hall 3:Offset and large printing equipment,package printing equipment,post press     

INFOPRINT 2014 was Successfully Held in Dongguan

Sponsored by Printing and Printing Equipment Industries Association of China （PEIAC） and organized by Print China magazine, the Seventeenth Beijing International Printing Information Conference （INFOPRINT 2014） was successfully held on 11th Dec. 2014 at Dongguan Exhibition International Hotel.