梅干菜和腌制雪菜细菌菌群多样性分析

为探讨传统腌制发酵蔬菜制品的菌群多样性,采用Illumina Miseq高通量测序技术,对宁波地区梅干菜半成品、成品和腌制雪菜成品进行分析。结果表明,梅干菜半成品、成品和腌制雪菜成品中相对丰度大于1.0%的菌属分别有8、13、5属,其中乳杆菌属（Lactobacillus）在3种产品中相对丰度均较高,分别为31.70%、9.51%、78.08%;假交替单胞菌（Pseudoalteromonas）仅在梅干菜半成品和腌制雪菜成品中检出,相对丰度为4.60%和1.81%;梅干菜半成品的主要优势菌属还有嗜盐单胞菌属（Halomonas）和梭状芽孢杆菌属（Clostridium IV）;梅干菜成品的主要优势菌属还有慢生芽孢杆菌属（Lentibacillus）、交替芽胞杆菌属（Alteribacillus）、嗜盐球菌属（Halococcus）、海单胞菌属（Oceanimonas）、枝芽孢菌属（Virgibacillus）、丛毛单胞菌属（Comamonas）、钠白菌属（Natrialba）和代尔夫特菌属（Delftia）等,相对丰度分别为11.85%、11.41%、9.64%、9.46%、8.86%、6.25%、5.21%和5.13%;其余优势菌属相对丰度在1.23%~2.60%之间。乳杆菌属是发酵蔬菜的主要优势菌,腌制方式、腌制阶段的蔬菜产品菌群多样性差异较大,干法腌制比泡制蔬菜产品的菌群结构和多样性更为复杂。本研究结果为传统腌制蔬菜微生物资源的挖掘及其工业化利用提供了理论参考。     

窖泥变质前后细菌群落差异分析

通过提取窖泥细菌总DNA,扩增16S r DNA构建基因文库,采用高通量测序的方法,对窖泥变质前后细菌群落进行分析。结果表明,窖泥变质前后细菌群落存在较大差异,共检测出16个门、174个种属。选取优势种属进行比较,窖泥变质前优势菌属为:动球菌属(22.63%)、芽孢杆菌属(12.95%)、紫单胞菌科(12.45%)、芽孢八叠球菌属(10.60%)等;窖泥变质后优势菌属为:芽孢八叠球菌属(61.34%)、嗜冷杆菌属(10.42%)、未分类菌属(5.45%)、子单胞菌属(5.02%)等。本研究初步揭示了窖泥变质前后细菌群落种类和数量的差异,为窖泥变质研究提供理论基础。     

基于变性梯度凝胶电泳和MiSeq高通量测序技术分析恩施地区腊肉的细菌多样性

以采集自湖北省恩施地区的5个腊肉样品为研究对象,采用聚合酶链式反应-变性梯度凝胶电泳(polymerase?chain?reaction-denatured?gradient?gel?electrophoresis,PCR-DGGE)与MiSeq高通量测序技术相结合的方法对其中所含微生物的多样性进行解析。PCR-DGGE结果表明,腊肉样品中的细菌主要为葡萄球菌属(Staphylococcus)、嗜冷杆菌属(Psychrobacter)、乳酸杆菌属(Lactobacillus)和假交替单胞菌属(Pseudoalteromonas);MiSeq高通量测序结果表明,腊肉中的细菌门主要为硬壁菌门(Firmicutes)和变形菌门(Proteobacteria),其平均相对含量分别为54.05%和44.28%,细菌属主要为葡萄球菌属(Staphylococcus)、嗜冷杆菌属(Psychrobacter)、假交替单胞菌属(Pseudoalteromonas)、环丝菌属(Brochothrix)、科贝特氏菌属(Cobetia)和不动细菌属(Acinetobacter),其平均相对含量分别为40.18%、24.02%、9.37%、8.53%、4.71%和2.31%;在分类操作单元(operational taxonomic units,OTU)水平上,发现252个核心OTU,累计平均相对含量高达74.22%。由此可见,PCR-DGGE和MiSeq高通量测序的结果综合分析得出恩施地区腊肉中的细菌主要为葡萄球菌属(Staphylococcus)、嗜冷杆菌属(Psychrobacter)和假交替单胞菌属(Pseudoalteromonas)。     

基于高通量测序分析虾夷扇贝柱菌群结构及腐败优势菌

为揭示虾夷扇贝柱的菌群结构及腐败优势菌,通过对宏基因组DNA进行高通量测序,分析虾夷扇贝柱新鲜和腐败样品的菌群结构,同时对可培养细菌菌群基因组DNA进行高通量测序,分析新鲜和腐败样品可培养细菌菌群的结构特征。结果表明,对宏基因组测序后获得的新鲜虾夷扇贝柱中细菌菌群多样性丰富,其中相对丰度较高的有Candidatus kuenenia（10.73%）、弧菌属（Vibrio,8.15%）、别弧菌属（Aliivibrio,7.38%）、芽孢杆菌属（Bacillus,7.20%）和未分类菌（unclassified,16.08%）;4?℃的腐败优势菌为发光杆菌属（Photobacterium,46.91%）、别弧菌属（36.82%）和假交替单胞菌属（Pseudoalteromonas,11.01%）,15?℃的腐败优势菌为发光杆菌属（46.97%）和别弧菌属（43.33%）,25?℃的腐败优势菌为发光杆菌属（41.94%）、别弧菌属（23.10%）、梭杆菌属（Fusobacterium,18.61%）和乳酸菌属（Lactobacillus,10.60%）。对可培养细菌菌群基因组测序后发现,新鲜样品可培养细菌菌群多样性大大降低,大部分为假交替单胞菌属（89.46%）,在25?℃腐败后,可培养的腐败优势菌为弧菌属（50.98%）、希瓦氏菌属（Shewanella,17.43%）和乳酸菌属（10.68%）。本研究揭示了虾夷扇贝柱新鲜及腐败样品的菌群结构特征及腐败优势菌,为进一步研究微生物对虾夷扇贝柱品质劣化的作用机制提供一定参考。     

基于Illumina MiSeq高通量测序分析清香型白酒酒糟微生物群落多样性

采用Illumina MiSeq高通量测序技术对清香型白酒酒糟中微生物多样性进行研究。结果表明,酒糟样品中细菌菌群的丰富度与多样性均大于真菌,细菌菌群归属于10个门、18个纲、31个目、66个科、86个属、124个种,真菌菌群归属于5个门、11个纲、20个目、33个科、49个属、66个种。优势细菌门（相对丰度≥1.0%）为变形菌门（Proteobacteria）、放线菌门（Actinobacteria）、厚壁菌门（Firmicutes）、拟杆菌门（Bacteroidetes）,优势细菌属为根瘤菌属（Rhizobium）、拉乌尔菌属（Raoultella）、谷氨酸杆菌属（Glutamicibacter）、红球菌属（Rhodococcus）、短波单胞菌属（Brevundimonas）、不动杆菌属（Acinetobacter）、假单孢菌属（Pseudomonas）、嗜冷杆菌属（Psychrobacter）、Devosia、嗜盐单胞菌属（Halomonas）、未分类-鼠杆菌属（unclassified-Muribaculaceae）、乳杆菌属（Lactobacillus）;优势真菌门为子囊菌门（Ascomycota）、毛霉门（Mucoromycota）及担子菌门（Basidiomycota）,优势真菌属为孢圆酵母属（Torulaspora）、伊萨酵母属（Issatchenkia）、横梗霉属（Lichtheimia）、根霉属（Rhizopus）、曲霉属（Aspergillus）、酵母属（Saccharomyces）。     

河套地区中温大曲细菌菌群多样性解析及其风味品质评价

为解析内蒙古河套地区中温大曲细菌类群及风味品质,该研究采用Illumina MiSeq高通量测序技术对采集自该地区的5份中温大曲细菌菌群结构及功能进行解析,同时采用电子鼻技术检测大曲风味品质,并通过纯培养技术对大曲中可培养芽孢杆菌（Bacillus）进行分离鉴定。结果表明,大曲中优势细菌属包括高温放线菌属（Thermoactinomyces）、乳杆菌属（Lactobacillus）、葡萄球菌属（Staphylococcus）、糖多孢菌属（Saccharopolyspora）、魏斯氏菌属（Weissella）、水杆菌属（Aquabacterium）、芽孢杆菌属（Bacillus）、嗜糖假单胞菌属（Pelomonas）、明串珠球菌属（Leuconostoc）、片球菌属（Pediococcus）和青枯菌属（Ralstonia）,累积平均相对含量为89.31%。其中Bacillus与有机硫化物、萜类物质、氮氧化物和芳香类物质之间存在显著正相关性（P<0.05）,Saccharopolyspora与有机硫化物呈显著正相关（P<0.05）。基因功能预测显示,大曲细菌菌群基因在氨基酸运输...     

腐乳中细菌群落结构与生物胺的相关性分析

通过高通量测序技术分析不同生物胺含量腐乳样品的细菌群落结构,测定相关环境因子,在此基础上探究菌群结构与生物胺含量之间的相关性。结果表明,腐乳中的优势菌门为厚壁菌门（Firmicutes）、变形菌门（Proteobacteria）、放线菌门（Actinobacteria）和拟杆菌门（Bacteroidetes）,四者的相对丰度之和在各样品中均达到91%以上;优势菌属包括芽孢杆菌属（Bacillus）、魏斯氏菌属（Weissella）、四联球菌属（Tetragenococcus）和假单胞菌属（Pseudomonas）;高低生物胺含量腐乳的组间主要差异菌属为芽孢杆菌属、魏斯氏菌属、假单胞菌属和赖氨酸芽孢杆菌属（Lysinibacillus）。相关性研究表明,总生物胺和总酸是影响细菌群落组成的主要因素（P<0.05）;生物胺含量主要与氨基酸态氮和pH等环境因子相关,腐乳样品的氨基酸态氮含量范围为0.62～0.86 g/100 g,pH变化范围在5.62～6.37之间;魏斯氏菌属、芽孢杆菌属和赖氨酸芽孢杆菌属丰度的增加与生物胺具有显著相关性;不动杆菌属（Acinetobacter）、丛毛单胞菌属（Comamonas）和四联球菌属与生物胺呈现负相关性。研究结果阐明了腐乳微生物群落结构与生物胺的关联性,为了解生物胺形成机制,有效控制生物胺和优化腐乳生产工艺提供了理论依据。     

酒饼叶对豉香型白酒酒饼细菌多样性影响研究

该文以广东省豉香型白酒的酒饼为研究对象,采用Illumina MiSeq高通量测序平台对不同比例酒饼叶生产的豉香型白酒酒饼中的细菌16S rDNA V3-V4区进行测序,分析豉香型白酒酒饼细菌菌群的多样性。结果表明,酒饼中细菌微生物群落构成比较稳定,90%以上分布在芽孢杆菌纲中,分别属于乳杆菌属、片球菌属和魏斯氏菌属等(相对丰度≥1%),不同比例酒饼叶对于豉香型白酒酒饼的细菌群落丰富度和多样性的影响各不相同;1%桂叶和8%假鹰爪叶生产的酒饼能够很好地提升其细菌菌群的丰富度和多样性;添加酒饼叶对乳杆菌属的相对丰度有抑制作用,桂叶和假鹰爪叶混合生产的酒饼可以大大提高魏斯氏菌属的相对丰度而降低片球菌属的丰度。该研究为优化豉香型白酒酒饼的生产发酵工艺和丰富酒曲微生物菌群数据库提供了理论依据。     

基于高通量测序分析不同保鲜冰处理对鲈鱼菌群组成与代谢功能的影响

目的：分析不同保鲜冰处理下鲈鱼微生物菌群变化及其与品质指标的相关性。方法：分别采用流化冰（slurry ice,SI）、酸性电解水冰（acidic electrolyzed water ice,AEWI）和碎冰（crushed ice,CI）对鲈鱼进行处理,测定鲈鱼的总挥发性盐基氮（total volatile basic nitrogen,TVB-N）含量与K值,并利用高通量测序技术对样品菌群结构和代谢功能进行比较分析。结果：SI和AEWI对鲈鱼均有一定的保鲜作用,其中SI抑制鲈鱼蛋白质和核苷酸降解速度的效果最显著。在冰藏前期（0～3 d）,AEWI与CI组样品中芽孢杆菌属（Bacillus）、乳球菌属（Lactococcus）、大洋芽孢杆菌属（Oceanobacillus）相对丰度明显高于SI组。冰藏中、后期（12～21 d）,SI组样品中假单胞菌属（Pseudomonas）、希瓦氏菌属（Shewanella）、摩替亚氏菌属（Moritella）、埃希氏杆菌属（Escherichia）和别弧菌属（Aliivibrio）相对丰度明显增加,AEWI组样品假单胞菌属（Pseudomonas）和不动杆菌属（Acinetobacter）相对丰度所占比例逐渐增大,CI组中不动杆菌属在21 d时达27.77%,成为造成鲈鱼贮藏中、后期腐败的主要菌属。主成分分析表明,冰藏方式及贮藏时间对鲈鱼菌群结构组成有较大影响。鲈鱼菌群中与TVB-N含量、K值呈正相关的有假单胞菌属、不动杆菌属、嗜冷杆菌属（Psychrobacter）和希瓦氏菌属,它们可能是鲈鱼冰藏期间的优势菌。CI和AEWI组鲈鱼细菌中参与氨基酸、脂质和碳水化合物代谢的相关基因相对丰度高于同一时期的SI组,这在一定程度上从细菌代谢水平解释了SI能延缓鲈鱼腐败的原因,为后期进一步从微生物抑制机制上优化鲈鱼贮藏保鲜技术提供了理论参考。     

基于Illumina Miseq技术比较不同地区传统发酵大豆制品细菌多样性

为深入挖掘和利用传统发酵大豆制品微生物菌种资源,采用Illumina Miseq高通量测序技术,分析了不同地区5类传统发酵大豆制品（纳豆、豆豉、大酱、霉豆渣、天贝）的细菌多样性。结果表明,5类产品菌落总数都在7.0~10.1 lg (CFU/g)之间。5类产品中共鉴定出相对丰度大于1%的细菌58个属,其中相对丰度大于10%的主要优势菌属有杆菌属（Bacillus）、枝芽孢菌属（Virgibacillus）、变形杆菌属（Proteus）、依格纳季氏菌属（Ignatzschineria）、四联球菌属（Tetragenococcus）、假单胞菌属（Pseudomonas）、不动杆菌属（Acinetobacter）、解脲芽孢杆菌属（Ureibacillus）、泛菌属（Pantoea）和片球菌属（Pediococcus）。主成分分析和聚类分析发现,纳豆1、纳豆2和豆豉1菌群结构相似,霉豆渣1和霉豆渣2菌群结构相似,霉豆渣3和大酱1菌群结构相似,天贝1与其他4类产品菌群结构差异较大。本研究结果为传统发酵大豆制品微生物资源的挖掘及其工业化利用提供了理论基础。     

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.     

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.     

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.     

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.     

Contribution of European research to risk analysis

The European Commission's, Quality of Life Research Programme, Key Action 1—Health, Food & Nutrition is mission-oriented and aims, amongst other things, at providing a healthy, safe and high-quality food supply leading to reinforced consumer confidence in the safety of European food. Its objectives also include the enhancing of the competitiveness of the European food supply. Key Action 1 is currently supporting a number of different types of European collaborative projects in the area of risk analysis. The objectives of these projects range from the development and validation of prevention strategies including the reduction of consumers risks; development and validation of new modelling approaches; harmonization of risk assessment principles, methodologies, and terminology; standardization of methods and systems used for the safety evaluation of transgenic food; providing of tools for the evaluation of human viral contamination of shellfish and quality control; new methodologies for assessing the potential of unintended effects of genetically modified (genetically modified) foods; development of a risk assessment model for Cryptosporidium parvum related to the food and water industries; to the development of a communication platform for genetically modified organism, producers, retailers, regulatory authorities and consumer groups to improve safety assessment procedures, risk management strategies and risk communication; development and validation of new methods for safety testing of transgenic food; evaluation of the safety and efficacy of iron supplementation in pregnant women; evaluation of the potential cancer-preventing activity of pro- and pre-biotic ('synbiotic') combinations in human volunteers. An overview of these projects is presented here.