山西河曲与内蒙古五原酸粥细菌类群结构及其基因功能的比较分析

该研究采用MiSeq高通量测序技术解析了河曲地区和五原地区酸粥的细菌群落结构差异，并探讨了其优势细菌属间的相关关系，最后对其细菌的基因功能进行了预测。结果发现，五原酸粥中的细菌丰度与多样性均显著高于河曲酸粥；所有酸粥样本共含有2个优势细菌门，分别为厚壁菌门(Firmicutes, 75.85%)和变形菌门(Proteobacteria, 23.31%);含有12个优势细菌属，分别为乳酸杆菌属(Lactobacillus,41.95%)、醋杆菌属(Acetobacter,14.10%)、芽孢杆菌属(Bacillus,6.72%)、梭菌属(Clostridium,6.69%)和戊糖片球菌(Pediococcus,5.36%)等，乳酸菌累计相对含量为59.19%;两个地区酸粥中Acetobacter和魏斯氏菌(Weissella)的含量存在显著差异。相关性分析显示，除Lactobacillus和肠球菌属(Enterococcus)间为显著负相关关系外，Clostridium与Weissella,乳球菌属(Lactococcus)与Acetobacter和Enterococcus以及气单胞菌属...     

液熏罗非鱼片加工过程中微生物群落的PCR-DGGE 分析

为弄清液熏罗非鱼片加工过程中微生物污染的源头,以进一步防控产品的微生物污染提供依据。应用基于细菌16S rDNA 的PCR-DGGE(PCR-denaturing gradient gel electrophoresis,PCR- 变性梯度凝胶电泳)技术分析液熏罗非鱼片主要加工关键环节的微生物群落结构,提取样品中的细菌总DNA,对细菌的16S rDNA 的V6～V8 区段进行PCR 扩增后,进行变性梯度凝胶电泳(DGGE),对DGGE 图谱进行微生物多样性分析,对主要条带进行序列分析并构建系统发生树。结果表明：10 个条带所代表的优势种很可能来源于以下几个属：巨型球菌属(Macrococus)、微球菌属(Micrococus)、肠道细菌属(Enterobacter)、假单胞菌属(Pseudomonas)、弧菌属(Vibrio)、突柄杆菌属(Prosthecobacter)、布特菌属(Buttiauxella),其中肠道细菌属、微球菌属、假单胞菌属和弧菌属细菌都具有使产品腐败的潜能。本研究表明：液熏罗非鱼片中呈现微生物多样性,PCR-DGGE 技术可用于研究液熏罗非鱼片加工过程中的微生物群落结构及变化,且具有一定的优越性。     

自然发酵腐乳中细菌多样性评价

以腐乳为研究对象,利用变性梯度凝胶电泳(polymerase chain reaction-denaturing gradient gel electrophoresis,PCR-DGGE)技术与Miseq高通量测序技术相结合的手段解析及评价细菌多样性,同时利用传统纯培养的方法对乳酸菌进行分离、纯化与鉴定。结果显示:腐乳中主要微生物为变形菌门(Proteobacteria,80.45%),其中绿脓杆菌属、不动杆菌属、鞘氨醇杆菌属、布丘氏菌属和草螺菌属细菌为优势细菌属。同时通过PCR-DGGE和纯培养发现腐乳中鉴定出7株乳酸菌,其中2株为戊糖片球菌(Pediococcus pentosaceus),1株为乳酸片球菌(Pediococcus acidilactici),4株为屎肠球菌(Enterococcus faecium)。由此可知腐乳中蕴含着丰富的微生物资源,同时屎肠球菌(Enterococcus faecium)为优势乳酸菌。     

乳酸菌肉品发酵剂的发酵特性研究

对从湘西泡菜水中分离出的5株植物乳杆菌(Lactobacillus planterum),1株肠系膜明串珠菌(Leuconostocmesenteroides),1株戊糖片球菌(Pediococcuspentosaceus),进行耐盐和亚硝酸盐能力、生长和产酸能力、硝酸盐还原和氨基酸脱羧能力的研究以及一系列生理生化实验来验证其在肉品发酵上的可用性。结果表明:植物乳杆菌(Lactobacillus planterum-A)和戊糖片球菌(Pediococcuspentosaceus-6)对食盐和亚硝酸盐均具有较好的耐受性,对蛋白质和脂肪不具有明显的分解作用,不具有硝酸盐还原酶和氨基酸脱羧酶活性,对大肠杆菌(Escherichia coli),沙门氏菌(Salmonellas spp.)等实验菌株有较好的抑制作用;70℃可致死。符合作为肉品发酵剂的要求,可作为下步实验的出发菌株。     

自然发酵风干肠中乳酸菌的分离与鉴定

以自然发酵风干肠为研究对象,分析了细菌总数和乳酸菌菌数的变化情况,结果表明,乳酸菌为风干肠发酵过程中的优势菌群.通过对风干肠中乳酸菌的分离鉴定,共分离出戊糖片球菌(Pediococcus pentosaceus)、乳酸乳球菌乳酸亚种(Lactococcus lactis subsp lactis)、短乳杆菌(Lactobacillus brevis)、弯曲乳杆菌(Lactobacillus curvatus)和发酵乳杆菌(Lactobacilus fermentum)5株乳酸菌.24h产酸速率测定结果表明,弯曲乳杆菌＞短乳杆菌＞乳酸乳球菌乳酸亚种＞戊糖片球菌＞发酵乳杆菌.     

原料乳中嗜冷菌的分离鉴定及Nisin,EDTA对其抑制作用

嗜冷菌是引起低温食品多种致害的主要原因之一.选用LA培养基从低温贮藏原料乳中分离出14株具有典型特性的嗜冷菌,并时的嗜冷菌进行了分离鉴定.通过形态学及生理生化特性的研究,对筛选的14株嗜冷菌进行了鉴定.一共得到9种嗜冷菌的菌种,分别是:气单胞菌属(Aeromonas)3株、黄杆菌属(Flavobacterium)1株、巴斯德氏菌属(Pasteurella)2株、节杆菌属(Arthrobacter)3株、短杆菌属(Brevibacterium)1株、片球菌属(Pediococcus)1株、乳球菌属(Lactococcus)1株,明串珠菌属(Trichococcus)1株,微球菌属(Micrococcus)1株.同时研究了不同浓度的Nisin,EDTA协同对牛乳中嗜冷菌的抑制作用,结果表明在牛乳中加入40ms/ks～100mg/kg的Nisin和等量的EDTA,对嗜冷菌有较好的抑制作用.     

新疆地区驴乳源优良乳酸菌发酵剂的筛选及菌株益生特性

从新疆哈密地区的15个驴乳样品中分离筛选出38株疑似乳酸菌,其中7株菌具有明显凝乳特性.经16S rRNA基因测序鉴定为乳酸片球菌(Pediococcus acidilactici)HL12-21、戊糖片球菌(Pediococcus pentosaceus)HL29-5、蒙氏肠球菌(Enterococcus mundt...     

发酵香肠中分离纯化的三株乳酸菌产酸特性研究

为了研究从发酵香肠中分离纯化的3株乳酸菌粪肠球菌(Enterococcus faecalis)、戊糖片球菌(Pediococcus pentosaceus)和肠膜明串珠菌(Leuconostoc mesenteroides)的产酸性能,对这3株菌在不同pH、温度、NaCl、NaNO2条件下的生长情况及产酸情况进行了测定。研究结果显示:这3株菌中,肠膜明串珠菌和戊糖片球菌的生长特性较好;粪肠球菌的生长特性虽不如肠膜明串珠菌和戊糖片球菌,但产酸能力最好,戊糖片球菌耐盐性最好、肠膜明串珠菌耐亚硝酸盐的特性最好;在不同温度和pH条件的测试中,肠膜明串珠菌的生长能力最好,粪肠球菌次之。这3株乳酸菌在发酵肉制品中均产生乳酸。总之,这3株菌均具有用于发酵制备乳酸的能力。     

UHT杀菌乳中耐热菌的分离与鉴定

采用2种不同的细菌富集方式对5种不同品牌UHT（Ultra High Temperature）杀菌牛乳中残留的细菌进行了分离，得到18株细菌，并对它们进行了耐热性和细菌学鉴定试验。结果表明，18株细菌全部耐热，初步将其归为3个属。其中，片球菌属10株，均为戊糖片球菌，肠球菌属3株，1株类鸟肠球菌，2株粪肠球菌；微球菌属5株，1株嗜冷微球菌，4株变异微球菌。虽然这些耐热菌均为非致病菌，但是遇到适宜条件，这些耐热菌会生长繁殖，导致乳产品在贮藏和流通过程中发生变质。     

阿尤恩地区自然发酵牛乳中乳酸菌分离鉴定及多样性研究

为了分离、保藏自然发酵乳中乳酸菌菌种,丰富自然发酵乳中乳酸菌多样性信息。本文采用传统的纯培养分离方法和宏基因组16S rRNA基因测序技术对阿尤恩地区自然发酵牛乳的乳酸菌多样性进行研究。纯培养结果表明:5份自然发酵牛乳中共分离出111株乳酸菌,鉴定为5个属10个种,其中Lactococcus lactis,占总分离株的45.95%;宏基因组测序结果发现,5份样品中的细菌归属于8个门、66个属和131个种,其中优势菌门为Firmicutes(69.63%),优势菌属为Lactococcus(31.80%),优势菌种同样为Lactococcus lactis(22.95%),另外还检测到含量较高的Streptococcus thermophilus(17.12%)、Lactobacillus helveticus(14.44%)等菌种。通过分析发现,Lactococcus lactis为阿尤恩地区5份自然发酵牛乳样品的优势菌种,并且自然发酵牛乳样品中细菌种类丰富,样品间细菌组成存在相似性和差异性。     

Changes in the composition of the bacterial flora on tray-packaged pork during chilled storage analyzed by PCR-DGGE and real-time PCR

In this study, a polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) was used to investigate the changes in the composition of the bacterial population of tray-packaged pork during chilled storage. Relative quantitative real-time PCR was further used to evaluate the predominant spoilage bacteria obtained from DGGE analysis for their relative amount to the total bacteria in meat samples. DGGE analysis of the V3 and V6-V8 regions of the 16S rRNA gene showed that Pseudomonas were the predominant bacterial species at the end of the monitoring period. Real-time PCR expressed as the ΔΔC(T) method showed that the average 2(-ΔΔC)(T) values increased continually during the storage period from less than 0.001 at day 0 to 4.438 at the end of the monitoring, which indicated that the proportions of Pseudomonas within the total bacteria in meat samples increased. Both methods confirmed that Pseudomonas was the predominant spoilage bacteria. Practical Application: This study uses new techniques to identify bacteria in fresh retail pork and to follow changes in the bacterial population during 12 d refrigerated storage. Pseudomonads were found to increase with storage time, becoming the dominant flora after 12 d.     

不同贮存期高温大曲中乳酸菌的多样性及其耐受性分析

该研究对不同贮存期高温大曲中乳酸菌的数量、群落结构与多样性的变化规律进行分析。 结果表明,高温大曲贮存过程中,乳酸 菌的数量由完成制曲时（0 d）的1.7×104 CFU/g增长至30 d时的1.4×105 CFU/g,最终贮存至120 d时乳酸菌数量达到最大值（7.6×105 CFU/g）。 通过高通量测序,从大曲中共鉴定出5个属,分别为乳杆菌属（Lactobacillus）、乳球菌属（Lactococcus）、肠球菌属（Enterococcus）、片球 菌属（Pediococcus）、双歧杆菌属（Bifidobacterium）,其中格氏乳球菌（Lactococcus garvieae）在各贮存期30 d、60 d和120 d大曲乳酸菌中 占比最多,分别为66.31%、47.75%和47.18%。 通过富集培养和平板分离,从高温大曲中共分离、鉴定出18株乳酸菌,对其中6株乳酸的 生长曲线、耐温性、耐酸性和耐乙醇能力进行了测定,其中菌株pH4-3能耐受47 ℃高温,菌株JD-1能耐受酒精度8%vol,并且二者均耐 酸（pH 3.5）。     

真空包装冷却猪肉冷藏过程中菌相变化

应用传统微生物培养和PCR-DGGE方法研究真空包装冷却猪肉4℃贮藏过程中的菌相变化。细菌培养计数结果表明,乳酸菌生长迅速,在贮藏后期即超过了细菌总数值。DGGE结合16S rRNA基因序列分析结果表明,贮藏初期肉中初始菌相较复杂,贮藏末期主要是漫游球菌、肉食杆菌、乳杆菌、乳球菌和热死环丝菌成为优势腐败菌。     

冷却牦牛肉贮藏过程中优势菌的 PCR-变性梯度凝胶电泳分析

应用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术,研究托盘保鲜膜包装的冷却牦牛肉在4℃贮藏过程中的微生物多样性及其动态变化.直接从样品中提取细菌总的DNA,采用降落PCR扩增16S rDNA的V3可变区序列,再通过DGGE得到动态变化的指纹图谱,并对主要条带进行测序分析.结果表明:检测到的优势腐败菌为Pseudomonas sp.(假单胞菌)、Lactococcus sp.(乳球菌)、Acinetobacter sp.(不动杆菌)、Brochothrix thermosphacta(热死环丝菌)、Enterobacteriaceae bacterium(肠杆菌科细菌),此外还检测到Uncultured Citrobacter sp.(非培养的柠檬酸杆菌)和Staphylococcus sp.(葡萄球菌)     

PCR-DGGE法结合传统技术研究4℃冷链贮运条件下三文鱼菌相变化

为研究三文鱼在冷链贮运4 ℃条件下的细菌腐败机制,运用聚合酶链式反应-变性梯度凝胶电泳（polymerase chain reaction-denaturing gradient gel electrophoresis,PCR-DGGE）技术、传统鉴定技术以及PCR技术分析了4 ℃冷链贮运条件下的三文鱼中腐败菌菌相变化规律,并通过产量因子测得各优势菌株致腐能力,从而确定特定腐败菌。DGGE指纹图谱显示,贮藏期间微生物多样性降低,假单胞菌属和希瓦氏菌属条带亮度却逐渐提高。这表明这两个属的微生物在三文鱼冷藏期间逐渐成为优势菌。通过分离和鉴定贮藏末期腐败三文鱼的优势腐败菌,本实验得到5 株优势腐败菌,分别为麦芽糖肉食杆菌（Carnobacterium maltaromaticum LMA28）、丁香假单胞菌（Pseudomonas syringae pv. syringae B728a）、荧光假单胞菌（Pseudomonas fluorescens SBW25）、肉食杆菌（Carnobacterium sp. WN1359）和波罗的海希瓦氏菌（Shewanella baltica OS678）。将这5 株纯培养的腐败菌分别接种到无菌三文鱼中并冷藏一定时间后,各腐败菌的挥发性盐基氮（total volatile basic nitrogen,TVB-N）产量因子分别为1.26×10－7、1.25×10－7、1.36×10－7、1.08×10－7 mg TVB-N/CFU和1.03×10－7 mg TVB-N/CFU。这5 株腐败菌对三文鱼致腐败能力的顺序依次为荧光假单胞菌SBW25＞麦芽糖肉食杆菌LMA28＞丁香假单胞菌B728a＞肉食杆菌WN1359＞波罗的海希瓦氏菌OS678。     

Bacterial diversity dynamics of traditional Turkish Ezine Cheese as evaluated by PCR‐DGGE and SSCP analysis

The aim of this study was to evaluate the bacterial ecosystem of milk and Ezine cheese by PCR amplification of the V3 region of the bacterial 16S rRNA gene followed by denaturing gradient gel electrophoresis (DGGE) and by monitoring the bacterial diversity dynamics using PCR single‐strand conformation polymorphism (SSCP) analysis. PCR‐DGGE analysis revealed that 17 different bands and strains belonging to the Lactococcus lactis group and Streptococcus thermophilus were predominant during manufacturing and ripening. SSCP analysis revealed that the bacterial profiles of the two cheese samples were similar.     

Diversity of the Predominant Spoilage Bacteria in Water-Boiled Salted Duck during Storage

Abstract: The spoilage microbiota in water-boiled salted duck during storage at 4 °C was determined using culture-dependent and independent methods. Analysis of the denaturing gradient gel electrophoresis (DGGE) patterns of PCR amplicons targeting the V3 region of the 16S rDNA and sequencing of the bands allowed profiling of the microbiota present in the duck. Community DNA extracts were prepared directly from water-boiled salted duck and from culturable bacterial fractions harvested from both MRS and PCA media. The spoilage bacteria mainly consisted of Staphylococcus saprophyticus, Macrococcus caseolyticus, Weissella, Halomonas sp. or Cobetia sp., and Exiguobacterium sp. based on sequencing and homology search of the DGGE bands. It appeared that both the bacterial counts and diversity increased during storage time. By plating method, bacterial counts in MRS agar increased from 10⁴ to 10⁸ CFU/g from day 1 to 10, while total bacterial counts in PCA agar reached 10⁹ CFU/g after 10 d. Total of 14 strains isolated from PCA and MRS agar were identified as M. caseolyticus (2), S. saprophyticus (7), S. sciuri (1), W. paramesenteroides (2), and W. confusa (2) by 16S rDNA sequencing. The identification of the spoilage-related microbiota is helpful to better understand the bacteria ecology in water-boiled salted duck and may lead to the discovery of appropriate preservation strategies.     

冷藏大菱鲆细菌组成变化和优势腐败菌

对大菱鲆(Scophthalmus maximus)在0、3、7、10℃冷藏过程中的细菌菌相进行定性和定量分析。研究发现新鲜大菱鲆细菌菌相比较复杂,分离得到114株细菌,其中革兰氏阴性菌和革兰氏阳性菌分别占68.4%和21.1%,优势菌为肠杆菌、侵肺巴斯德菌、拟态弧菌、假单胞菌属和腐败希瓦氏菌,比例分别为14.9%、13.2%、12.3%、10.5%和8.8%。冷藏过程中,细菌菌相逐渐单一,腐败希瓦氏菌和假单胞菌增长显著,0、3、7、10℃冷藏至较好品质期终点时,腐败希瓦氏菌比例分别为42.2%、39.0%、57.1%和56.3%,平均比例为50.2%;其次是假单胞菌,比例分别为28.0%、23.7%、25.5%和31.0%,平均比例为27.2%。0、3、7、10℃冷藏至货架期终点时,腐败希瓦氏菌比例分别为53.9%、50.0%、57.4%和47.6%,平均比例为52.3%;假单胞菌属比例分别为25.5%、25.0%、41.0%和15.9%,平均比例为26.5%。由此可得出大菱鲆在0～10℃冷藏过程中的优势腐败菌是腐败希瓦氏菌,其次是假单胞菌。     

The microbial community in a 2,4-dinitrophenol-digesting reactor as revealed by 16S rDNA gene analysis

The microbial community of a 2,4-dinitrophenol-digesting reactor was investigated using different molecular biological techniques based on 16S rDNA gene sequences. A PCR-denaturing gradient gel electrophoresis (DGGE) analysis of the bacterial community in the reactor showed that one strong and five minor bands were observed in the DGGE profile. The results of excising and sequencing DGGE bands suggested that members of Rhodococcus, Nocardioides, and Nitrospira species were present in the reactor. Partial sequencing of cloned 16S rDNAs revealed diversity among the six main divisions--the alpha, delta subclasses of Proteobacteria, Nitrospira, Cytophagal Flexibacter/Bacteroides, Verrucomicrobia, and Actinobacteria--in the reactor. Two cloned sequence types were not closely affiliated with any described bacterial divisions. The isolation and phylogenetic analysis of 2,4-DNP-degrading bacteria from the reactor revealed that isolated strains were classified into two types of bacteria having different 16S rDNA sequences. One of these strain types was identified as a relative of Rhodococcus koreensis, and the other was identified as a relative of Nocardioides simplex FJ21-A.