PCR-DGGE分析鲜切莲藕冷藏过程中的菌相变化

为明确鲜切莲藕的微生物腐败机制,基于PCR-DGGE技术分析其冷藏过程中的菌相变化。鲜切莲藕样品于4℃冷藏,每间隔3 d取出,PCR扩增其16S r DNA的V3高变区,将产物进行变性梯度凝胶电泳(DGGE),并对优势条带进行回收和测序。结果表明,冷藏过程中鲜切莲藕的菌相结构发生明显变化,冷藏6~9 d为其腐败的关键时间点,其中假单胞菌属、伯氏致病杆菌、克雷伯氏肺炎杆菌为主要腐败菌,尤其是假单胞菌属在冷藏后期最易增殖,逐渐成为优势腐败菌,可作为鲜切莲藕产品腐败评价或快速检测的重要指示菌。     

冷鲜鸡胸肉中2种优势腐败菌的分离鉴定

以冷鲜鸡胸肉为对象,研究在4℃冷藏条件下腐败菌菌相变化,并对其优势腐败菌进行分离鉴定。4℃下冷藏开始时,冷鲜鸡胸肉中假单胞菌属(Pseudomonas)和环丝菌属(Brochothrix)菌落总数之和仅占细菌总数的24%。随着冷藏时间的延长,两菌属迅速增长繁殖,于第2天成为优势腐败菌群,并于第(2~8)天持续占据优势。对优势菌落进一步分离纯化得到两株优势腐败菌CM1、CM2。通过传统微生物生理生化鉴定和Biolog GenⅢ表型测试对2菌株进行初步鉴定,并进一步结合16S r DNA序列分析建立系统发育树,确定CM1菌株与Pseudomonas deceptionensis、CM2菌株与Brochothrix thermosphacta亲缘关系最近。该研究结果可为冷鲜鸡胸肉的贮藏保鲜及抑菌研究提供理论依据。     

冷藏期间鲜切莲藕的微生物多样性变化规律分析

为探究鲜切莲藕在冷藏期间微生物多样性的变化规律，明确鲜切莲藕冷藏期间的主要微生物种类及其生长规律，以4℃贮藏的鲜切莲藕为研究对象，分别对贮藏0、3、6、9、12 d的鲜切莲藕中真菌ITS和细菌16S rDNA序列进行高通量测序分析。结果表明，冷藏期间鲜切莲藕中微生物，共鉴定出真菌2个门、19个属，细菌2个门、5个属。在不同冷藏阶段，鲜切莲藕中的微生物存在差异。在属水平上，冷藏0 d时优势真菌为陶氏菌属，3～12 d冷藏阶段的优势真菌为长垣菌属，且长垣菌属在冷藏12 d时极显著富集；冷藏0～9 d优势细菌为不动杆菌属，而冷藏12 d时优势细菌为假单胞菌属，且假单胞菌属在冷藏12 d时极显著富集。通过研究初步明确了冷藏期间鲜切莲藕的微生物菌群结构和变化规律，为鲜切莲藕保鲜技术和质量控制研究提供参考。     

鲜切莲藕腐败的荧光检测方法

针对传统微生物卫生指标法检测果蔬产品腐败的局限性,研究利用腐败指示菌的荧光检测表征鲜切莲藕样品腐败的方法。鲜切莲藕样品于4 ℃冷藏,每间隔3 d取出检测其微生物菌落总数和指示菌自发荧光检出的相关性,分析指示菌荧光强度与微生物数量的关系并优化荧光检测条件。结果显示:鲜切莲藕冷藏第6天菌落总数为6.4×104 CFU/g,达到腐败标准,而自冷藏第6天开始,莲藕悬液培养后均能检测到荧光,表明荧光检出时间与样品腐败时间点一致;激光共聚焦荧光检测显示样品悬液的荧光强度随其冷藏时间的延长而增强,随其稀释度的增加而降低,表明荧光强度与微生物数量间呈正相关;最终确定该荧光检测方法的最优条件为样品悬液稀释10倍、接种量5 μL,对随机10个样品进行检测验证,准确率达90%,操作简单、便捷可行。     

鲜肉粽货架期致腐微生物研究

对影响鲜肉粽货架期的主要微生物进行菌相分析研究,结果表明在低于25℃的储藏条件下,影响鲜肉粽货架期的优势腐败菌是葡萄球菌和芽孢杆菌。鲜肉粽在货架存放0～2d,微生物未检出;存放3d时,细菌总数呈上升趋势,主要优势菌是葡萄球菌;存放5d时,芽孢杆菌占优势地位,霉菌增长缓慢;在37℃存放条件下,优势腐败菌为霉菌。通过对优势腐败微生物的分离,综合菌落形态观察结果,生理生化试验和16S rDNA测序,经鉴定优势腐败细菌为表皮葡萄球菌(S.epidermidis)、枯草芽孢杆菌(Bacillus subtilis)和短小芽孢杆菌(Bacillus pumilus)。结合霉菌菌落观察结果和培养特性,初步确定霉菌为曲霉和青霉。     

低温下鲜切莲藕菌相分析及货架期评价

采用不同选择性培养基分析真空包装鲜切莲藕的菌相组成,并采用菌落形态、显微形态观察及生理生化方法予以鉴定;建立Gompertz模型揭示莲藕主要腐败微生物在4℃恒温贮藏过程中的变化规律;基于感官分析,确定低温条件下鲜切莲藕货架期的微生物限量。结果发现,真空包装鲜切莲藕中主要菌相为乳酸菌属、肠杆菌科、假单胞菌属、酵母菌属,实验鉴定出乳酸菌属、肠杆菌科、假单胞菌属、酵母菌属各2株;经菌落形态、显微形态观察及生理生化鉴定,均符合相关科属特性。整个贮藏过程中,乳酸菌、肠杆菌、假单胞菌和酵母菌的数量变化趋势基本一致,其中乳酸菌延滞期最短,假单胞菌最大比生长率最高,酵母菌数量优势相对较大。该莲藕产品的货架期为10d左右,所对应的微生物数量级约107CFU/g。     

冷却猪肉中优势腐败菌致腐能力研究

为明确冷却猪肉中优势腐败菌对其腐败的作用程度,本研究监测冷却猪肉冷藏过程中菌落总数和优势腐败菌的变化情况,并接种假单胞菌、气单胞菌、肠杆菌和热杀索丝菌于冷却猪肉中,分组检测尸胺含量在冷却猪肉冷藏过程中的变化。实验表明,优势腐败菌在冷却猪肉冷藏第5d后显著上升(p<0.05),与尸胺含量变化趋势相似,同时接种多种腐败菌与尸胺含量极显著相关(p<0.01),表明优势腐败菌的协同效应对冷却猪肉腐败具有重要作用。     

冷藏大口黑鲈鱼片品质变化规律及优势腐败菌致腐能力分析

本研究的目的为分析大口黑鲈鱼片冷藏过程中品质的变化规律和确定贮藏过程中的优势腐败菌及其致腐能力。以新鲜鲈鱼为原料,通过测定鲈鱼肉的质构特性、菌落总数、挥发性盐基氮(Total volatile basic nitrogen, TVB-N)和K值等指标,对冷藏过程中鱼片的品质和优势腐败菌及其致腐能力进行了分析,并通过16S rRNA序列鉴定分析各腐败菌的种属。质构特性、菌落总数、TVB-N 总挥发性盐基氮(total volatile basic nitrogen, TVB-N)、K值等品质相关指标的测定结果表明,随着贮藏时间的延长,鱼片的弹力、咀嚼性和胶着性均呈缓慢下降的趋势;菌落总数逐渐增长;TVB-N在贮藏前7 d变化不大,在第7 d后急速上升;K值在第7 d时超过了60%,达到了初期腐败。采用选择性培养基获得鲈鱼腐败期的特定菌落,结合菌落形态观察、16S rDNA与革兰氏染色技术,确定所属菌株属种,并评价各菌株的致腐能力。筛选出T2、C1和C2 三3株菌为4℃贮藏鲈鱼片的疑似优势腐败菌。16S rDNA鉴定这3株菌分别为希瓦氏菌、假单胞菌和普罗维登斯菌。根据接种这3三株腐败菌的大口黑鲈鱼片的TVB-N产量因子(YTVB-N/cfu)的评价结果,确定这三3株菌的致腐能力为假单胞菌>希瓦氏菌>普罗维登斯菌。假单胞菌为4℃贮藏的鲈鱼片腐败的优势菌。     

应用表型和基因型分析28℃鲜湿米粉中主要优势菌

为研究鲜湿米粉中的优势菌,本文采用传统微生物培养方法和16S r DNA技术对28℃条件下鲜湿米粉的腐败菌进行分离鉴定,并了解鲜湿米粉贮藏期间的优势菌群与主要优势菌。结果表明,鲜湿米粉在28℃条件下,1 d后其菌落总数超过限量要求,2 d后感官出现明显腐败变质;经表型鉴定表明,其优势菌群包括肠杆菌科、乳酸菌、芽孢杆菌属,经基因型鉴定表明,肠杆菌属是导致鲜湿米粉腐败变质的主要优势菌。随着贮藏时间的延长,肠杆菌科占所有菌相的51.3%,是造成鲜湿米粉腐败变质的主要优势菌。     

冷藏中国对虾优势腐败菌的分离鉴定及致腐能力分析

为研究中国对虾在4℃的优势腐败菌及其致腐能力,通过培养基法筛选获得单一菌株,对冷藏中国对虾的优势腐败菌以16S rDNA技术结合细菌生理生化试验鉴定中国对虾的优势腐败菌。将各腐败菌接种于无菌中国对虾中,分析中国对虾的TVB-N、pH、菌落总数等指标,对各菌株进行致腐能力的测定。结果表明,分离的4株中国对虾腐败菌,分别为荧光假单胞菌(Pseudomonas fluorescens)、威斯康星米勒菌(Moellerella wisconsensis)、地衣芽孢杆菌(Bacillus licheniformis)和腐败希瓦菌(Shewanella putrefaciens);致腐能力由高至低依次为荧光假单胞菌>腐败希瓦菌>威斯康星米勒菌>地衣芽孢杆菌;确定在4℃冷藏下荧光假单胞菌(Pseudomonas fluorescens)是中国对虾的优势腐败菌。     

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.     

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.     

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.     

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