食品和水中大肠埃希氏菌验证方法的比较

目的探讨IMViC检测实验培养基在大肠菌群验证实验中的适用性,优选相对快速确证为大肠菌群的培养基。方法比较IMViC检测实验培养基与国标法中验证实验所用培养基测定大肠埃希氏菌以及食品、生活饮用水、天然饮用矿泉水中大肠菌群不同时段的结果,对比所有样本确证为大肠菌群所需时间。结果测定大肠埃希氏菌以及食品、生活饮用水、天然饮用矿泉水中大肠菌群时, IMViC检测实验培养基6 h测定结果与国标法最终结果均无显著性差异(P0.05)。结论 IMViC检测实验培养基适用于大肠菌群验证试验,可根据实验室检测需求选择适用于大肠菌群快速检测实验的培养基。     

Taqman探针荧光PCR法检测食品中的大肠埃希氏菌O145

针对大肠埃希氏菌O145的O抗原基因簇的wck D基因的特异性序列设计引物和Taqman探针,建立检测大肠埃希氏菌O145的荧光PCR方法,对其灵敏度、特异性进行验证,并将其用于食品样品的检测。结果表明,本研究中的方法可实现对大肠埃希氏菌O145的特异性扩增,其它27株非O145大肠埃希氏菌和20株非大肠埃希氏菌细菌的菌株均无扩增;检测的灵敏度可达165拷贝/反应;339份食品样品EC肉汤增菌后用本荧光PCR法进行检测,检出大肠埃希氏菌O145阳性31份,阳性率为9.1%。实验结果表明,本研究成功建立了可用于食品中大肠埃希氏菌O145的Taqman探针荧光PCR方法,食品样品采用EC肉汤增菌24 h、热裂解法提取核酸,增菌后检测所需时间由至少3 d~7 d缩短为仅需2 h~3 h,食品检测全过程仅需约28 h,经证实本方法特异性强、操作简便,为食品中大肠埃希氏菌O145提供了一种的快速检测手段。     

酶底物法快速检测食品中的大肠埃希氏菌

目的将水质检验的酶底物法用于食品检验,探索酶底物法是否能快速检测食品中的大肠埃希氏菌。方法测定食品样品,比较酶底物法与传统鉴定法结果的一致性。结果传统方法检出24份阳性和8份阴性,酶底物法检出25份阳性和7份阴性。实验结果进行X~2检验,结果无显著性差异(X~2=0.087,P0.05)。32份样品MPN值结果一致,8份样品MPN值有差异,其中7份样品酶底物法的MPN值大于传统法。结论酶底物法可用于食品中大肠埃希氏菌的快速检测,具有快捷简便、特异性高的优势。     

侧向流动型重组酶聚合酶扩增技术检测食品中大肠埃希氏菌O157

研究内容基于侧向流动型重组酶聚合酶扩增(Lateral flow-based recombinase polymerase amplification,LF-RPA)技术原理,建立大肠埃希氏菌O157(Escherichia coli O157)的LF-RPA快速检测方法。应用该方法对3株大肠埃希氏菌O157和27株非大肠埃希氏菌O157进行检测,结果表明该方法具有良好的特异性。分别以10倍浓度梯度稀释插入靶基因序列的质粒pUC57-rfbE和大肠埃希氏菌O157菌液DNA作为模板进行LF-RPA检测。结果表明LF-RPA方法可检出1.0×10²拷贝/μL的pUC57-rfbE质粒和1.2×10⁴ cfu/mL的大肠埃希氏菌O157菌液。以大肠埃希氏菌O157污染食品样品,增菌4 h后,大肠埃希氏菌O157初始浓度为1.2×10¹ cfu/mL的食品增菌液可以被LF-RPA检出阳性信号。该研究建立的方法可快速、准确、灵敏地检测食品中大肠埃希氏菌O157,无需专业设备,在食品生产企业和基层检测机构具有一定的应用前景。     

荧光型重组酶聚合酶扩增技术检测食品中大肠埃希氏菌O157

研究基于荧光型重组酶聚合酶扩增(exo-RPA)技术原理,建立了一种大肠埃希氏菌O157的快速检测方法。应用该方法对多种大肠埃希氏菌O157菌株和非大肠埃希氏菌O157菌株的检测结果表明,该方法的荧光探针和引物具有良好的特异性。对于梯度稀释的插入靶基因序列的质粒pUC57-rfbE和大肠埃希氏菌0157的检测,大肠埃希氏菌O157 exo-RPA检测灵敏度分别可以达到10~2 copies/μL和2.1×10~4 cfu/mL。对食品样品中大肠埃希氏菌0157的检测,exo-RPA方法显示出了良好的稳定性。并且食品样品中目标菌经4 h增菌后,该方法的灵敏性可以满足对食品样品中初始浓度为2.1×10~1 cfu/mL的目标菌的检测。因为大肠埃希氏菌exo-RPA方法配套设备简便、廉价,所以该方法可以在各级检测机构中推广,并适用于食品生产到消费各环节中大肠埃希氏菌0157的即时检测。     

实时荧光定量PCR法快速检测食品中大肠埃希菌

目的应用实时荧光定量PCR技术,结合3~5 h的前增菌处理,建立食品中大肠埃希菌的快速、灵敏、定量的检测方法。方法以大肠埃希菌(ATCC 25922)为参考菌株,对培养基和培养温度进行优化,选择最佳的前增菌培养条件。将不同浓度的参考菌株和样品分别接种前增菌液中培养3~5 h。采用Triton-X 100提取增菌后的DNA,实时荧光定量PCR扩增大肠埃希菌特异性片段。所得Ct与对应的原始(增菌前)参考菌株的浓度,建立标准曲线,计算样品中大肠埃希菌的数量。结果纯培养模式下,经过3、4和5 h的前增菌后,标准曲线具有很好的线性,r2分别为0.996、0.992和0.991,对应的检测限为136、14和1.4 cfu/100 ml;含杂菌培养模式下,NB和EC肉汤42.0℃增菌4 h后,建立的标准曲线r2分别为0.972和0.978。在不同食品中该方法的加标回收率为74.0%~174.0%。结论 3~5 h的前增菌实时荧光定量PCR方法可以快速、灵敏、定量地检测食品中活的大肠埃希菌。     

智舌检测出血性大肠埃希氏菌O157:H7和金黄色葡萄球菌

目的:探究一种快速检测出血性大肠埃希氏菌O157:H7、金黄色葡萄球菌的新方法。方法:基于多频大幅脉冲伏安法的智舌检测微生物生长过程中营养肉汤成分的综合信息,并结合主成分分析方法和簇类的独立软模式识别技术,对检测得到的数据进行分析。结果:在智舌的钯电极1 Hz频率段检测出血性大肠埃希氏菌O157:H7、金黄色葡萄球菌和1%葡萄糖营养肉汤区分效果较好,通过独立软模式识别技术能很好地判别、区分有菌生长的培养基和纯培养。结论:智舌能够有效检测、区分两种菌及培养基,有望成为检测出血性大肠埃希氏菌O157:H7、金黄色葡萄球菌的一种新技术。     

广州市蔬菜类凉拌菜中微生物污染状况调查及大肠埃希氏菌耐药现象研究

;目的 了解广州市蔬菜类凉拌菜的微生物污染状况及大肠埃希氏菌耐药情况,为食源性疾病的预防和控制提供科学依据。方法：从广州市的超市、集贸摊点采集300份凉拌菜,依据GB 4789-2016进行微生物检验,用VETIK 对检出的大肠埃希氏菌进行药敏试验,PCR法检测致泻性大肠埃希氏菌毒力基因。结果：300份样品中大肠菌群检出率为41.7%,大肠埃希氏菌检出率为6.7%,金黄色葡萄球菌检出率为2.0%,未检出沙门氏菌。药敏结果显示大肠埃希氏菌对头孢唑啉的抗性最高(40.0%)、其次为氨苄西林(30.0%),对其它受试抗生素不具有耐药性,且无多重耐药菌株。20株大肠埃希氏菌中检出3株致泻大肠埃希氏菌。结论：广州地区蔬菜类凉拌菜中存在较为严重的微生物污染,食源性大肠埃希氏菌具有一定的耐药现象,应加大卫生监督力度,保障食品安全。     

3种方法检测食品中致泻大肠埃希氏菌检测能力验证结果分析

目的提高食品中致泻大肠埃希氏菌的检测能力,促进实验室检测能力的提高。方法参照GB4789.6-2016《食品微生物学致泻大肠埃希氏菌检验》方法进行检测,采用VITEK 2 Compact全自动细菌鉴定系统进行生化鉴定、血清学鉴定,对可疑菌落进行普通PCR确证试验。同时使用多重实时荧光PCR法以及基质辅助激光解析电离飞行时间质谱(matrix-assisted laser desorption ionizationtime-of-flight mass spectrometry,MALDI-TOF-MS)分析鉴定可疑菌落。结果国标法和多重实时荧光PCR法能准确鉴定出目标菌,MALDI-TOF-MS可以检测大肠埃希氏菌,但无法区分致泻大肠埃希氏菌和肠出血性大肠埃希氏菌。结论 3种方法各有优劣,同时使用,综合判断,能确保试验结果准确快速。     

傅立叶变换红外光谱技术对6种致病性大肠埃希氏菌快速分型的研究

目的建立肠致病性大肠埃希氏菌(Enteropathogenic E.coli,EPEC)、肠产毒性大肠埃希氏菌(Enterotoxigenic E.coli,ETEC)、肠侵袭性大肠埃希氏菌(Enteroinvasive E.coli,EIEC)、肠黏附性大肠埃希氏菌(Enteroadhesive E.coli,EAEC)、肠出血性大肠埃希氏菌(Enterohmorrhagic E.coli,EHEC)、弥散粘附性大肠埃希氏菌(Diffusely adherent E.coli,DAEC)6种致病性大肠埃希氏菌的傅立叶变换红外光谱(Fourier transform infrared spectroscopy,FT-IR)数据库及FT-IR分型方法。方法应用FT-IR技术对6种致病性大肠埃希氏菌进行指纹图谱数据采集并对其进行基线校正、归一化等处理,应用化学计量学方法对光谱数据进行分析。结果本研究建立了6种致病性大肠埃希氏菌的FT-IR光谱数据库,实现了对6种可疑目标大肠埃希氏菌鉴定;建立了主成分分析(principal component analysis,PCA)和分级聚类分析(hierarchical cluster analysis,HCA)2种模型,成功实现了对6种致病性大肠埃希氏菌的快速分型。结论 FT-IR分析方法快速、简便、易操作,结果重现性好,是一种鉴定6种致病性大肠埃希氏菌的有效方法。     

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.     

An internet compendium of analytical methods and spectroscopic information for monomers and additives used in food packaging plastics

An internet website (http://cpf.jrc.it/smt/) has been produced as a means of dissemination of methods of analysis and supporting spectroscopic information on monomers and additives used for food contact materials (principally packaging). The site which is aimed primarily at assisting food control laboratories in the European Union contains analytical information on monomers, starting substances and additives used in the manufacture of plastics materials. A searchable index is provided giving PM and CAS numbers for each of 255 substances. For each substance a data sheet gives regulatory information, chemical structures, physico-chemical information and background information on the use of the substance in particular plastics, and the food packaging applications. For monomers and starting substances (155 compounds) the infra-red and mass spectra are provided, and for additives (100 compounds); additionally proton NMR are available for about 50% of the entries. Where analytical methods have been developed for determining these substances as residual amounts in plastics or as trace amounts in food simulants these methods are also on the website. All information is provided in portable document file (PDF) format which means that high quality copies can be readily printed, using freely available Adobe Acrobat Reader software. The website will in future be maintained and up-dated by the European Commission's Joint Research Centre (JRC) as new substances are authorized for use by the European Commission (DG-ENTR formerly DGIII). Where analytical laboratories (food control or other) require reference substances these can be obtained free-ofcharge from a reference collection housed at the JRC and maintained in conjunction with this website compendium.     

Chlorohydrins of bisphenol A diglycidyl ether (BADGE) and of bisphenol F diglycidyl ether (BFDGE) in canned foods and ready-to-drink coffees from the Japanese market

BADGE.2HCl and BFDGE.2HCl were determined in 28 samples of ready-to-drink canned coffee and 18 samples of canned vegetables (10 corn, 5 tomatoes and 3 others), all from the Japanese market. HPLC was used as the principal analytical method and GCMS for confirmation of relevant LC fractions. BADGE.2HCl was found to be present in one canned coffee and five samples of corn, BFDGE.2HCl in four samples of canned tomatoes and in one canned corn. No sample was found which exceeded the 1mg/kg limit of the EU for the BADGE chlorohydrins. However the highest concentration was found for the sum of BFDGE.2HCl and BFDGE.HCl.H₂O at a level of 1.5mg/kg. A Beilstein test confirmed that all cans containing foods contaminated with BADGE.2HCl or BFDGE.2HCl had at lest one part coated with a PVC organosol.     

European priorities for research to support legislation in the area of food contact materials and articles

A strong science base is required to underpin the planning and decision-making process involved in determining future European community legislation on materials and articles in contact with food. Significant progress has been made in the past 5 years in European funded work in this area, with many developments contributing to a much better understanding of the migration process, and better and simpler approaches to food control. In this paper this progress is reviewed against previously identified work-areas (identified in 1994) and conclusions are reached about future requirements for R&D to support legislation on food contact materials and articles over the next 5 or so years.     

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.     

Textural Properties of Cold-set Gels Induced from Heat-denatured Whey Protein Isolates

A 9% whey protein (WP) isolate solution at pH 7.0 was heat-denatured at 80°C for 30 min. Size-exclusion HPLC showed that native WP formed soluble aggregates after heat-treatment. Additions of CaCl2 (10–40 mM), NaCl (50–400 mM) or glucono-delta-lactone (GDL, 0.4–2.0%, w/v) or hydrolysis by a protease from Bacillus licheniformis caused gelation of the denatured solution at 45°C. Textural parameters, hardness, adhesiveness, and cohesiveness of the gels so formed changed markedly with concentration of added salts or pH by added GDL. Maximum gel hardness occurred at 200 mM NaCl or pH 4.7. Increasing CaCl2 concentration continuously increased gel hardness. Generally, GDL-induced gels were harder than salt-induced gels, and much harder than the protease-induced gel.     

Influence des ions sur le pouvoir hydratant de l'ure e: e tude sur peau de porc ex vivo

This study deals with the influence of ions (NaCl and MgSO4) in a W/O emulsion containing 10% urea. Moisturization kinetics are assessed by corneometry on pig skin ex vivo. The formula's influence on urea penetration is measured by infrared spectrometry with an ATR device and the stripping method. Corneometry and spectroscopy were chosen to record simultaneously the hydratation levels and urea localization into superficial cell layers. Urea crystallization after evaporation of emulsions and aqueous solutions is described. Results show that urea does not hydrate nor penetrate when applied to the skin through an aqueous gel. In a W/O emulsion, sodium chloride increases the ability of urea to moisturize without improving penetration. In vitro urea crystallization is disturbed by sodium chloride or magnesium sulphate for solutions and emulsions. This stabilization by ions is correlated with good moisturization values. The stabilization of urea in the solute state provided by ions increases its water epidermal binding capacity without enhancing penetration.     

Occurrence of bisphenol-F-diglycidyl ether (BFDGE) in fish canned in oil

The levels of bisphenol-F-diglycidyl ether (BFDGE) were quantified as part of a European survey on the migration of residues of epoxy resins into oil from canned fish. The contents of BFDGE in cans, lids and fish collected from all 15 Member States of the European Union and Switzerland were analysed in 382 samples. Cans and lids were separately extracted with acetonitrile. The extraction from fish was carried out with hexane followed by re-extraction with acetonitrile. The analysis was performed by reverse phase HPL C with fluorescence detection. BFDGE could be detected in 12% of the fish, 24% of the cans and 18% of the lids. Only 3% of the fish contained BFDGE in concentrations considerably above 1mg/kg. In addition to the presented data, a comparison was made with the levels of BADGE (bisphenol-A-diglycidyl ether)analysed in the same products in the context of a previous study.