PCR-DGGE分析甘薯酸浆自然发酵过程中细菌多样性

以不同发酵阶段的甘薯酸浆为研究对象,采用变性梯度凝胶电泳技术（denaturing gradient gelelectrophoresis,DGGE）分析甘薯酸浆发酵过程中细菌的多样性。提取样品总DNA,聚合酶链式反应（polymerasechain reaction,PCR）扩增16S rDNA的高变区序列,再通过变性梯度凝胶电泳得到特异性条带并对其进行序列分析及比对。结果表明：甘薯酸浆自然发酵过程中,主要以厚壁菌门（Firmicutes）和变形菌门（Proteobacteria）的明串珠菌属（Leuconostoc）、不动杆菌属（Acinetobacter）、Reyranella属、泛菌属（Pantoea）、克罗诺杆菌属（Cronobacter）以及螺菌属（Spiriiium）的细菌为主;此外,还有拟杆菌门中黄杆菌属（Flavobacterium）以及蓝细菌中的色球藻属（Chroococci）的细菌参与其中。相似性分析结果表明,自然发酵甘薯酸浆中的菌群结构有很大相似性,其中最大相似度为88.5%,最小相似度为71.4%。     

Microbial Diversity of Type I Sourdoughs Prepared and Back‐Slopped with Wholemeal and Refined Soft (Triticum aestivum) Wheat Flours

The fermentation of type I sourdough was studied for 20 d with daily back‐slopping under laboratory and artisan bakery conditions using 1 wholemeal and 2 refined soft wheat (Triticum aestivum) flours. The sourdough bacterial and yeast diversity and dynamics were investigated by plate counting and a combination of culture‐dependent and culture‐independent PCR‐DGGE approach. The pH, total titrable acidity, and concentration of key organic acids (phytic, lactic, and acetic) were measured. Three flours differed for both chemical and rheological properties. A microbial succession was observed, with the atypical sourdough species detected at day 0 (i.e. Lactococcus lactis and Leuconostoc holzapfelii/citreum group for bacteria and Candida silvae and Wickerhamomyces anomalus for yeasts) being progressively replaced by taxa more adapted to the sourdough ecosystem (Lactobacillus brevis, Lactobacillus alimentarius/paralimentarius, Saccharomyces cerevisiae). In mature sourdoughs, a notably different species composition was observed. As sourdoughs propagated with the same flour at laboratory and artisan bakery level were compared, the influence of both the substrate and the propagation environment on microbial diversity was assumed.     

Selection of metabolic pathways for continuous hydrogen production under thermophilic and mesophilic temperature conditions in anaerobic fluidized bed reactors

This study evaluated the influence of hydraulic retention time (HRT) on hydrogen (H₂) production in anaerobic fluidized bed reactors at mesophilic (30 °C, AFBR-M) and thermophilic (55 °C, AFBR-T) temperatures. Reactors were fed sucrose-based synthetic wastewater (5000 mg chemical oxygen demand·L^?1) in the HRT of 8, 6, 4, 2, or 1 h. H₂ production rate increased from 67.8 ± 14.8 to 194.9 ± 57.0 ml H₂·h^?1 L^?1 (AFBR-T) and from 72.0 ± 10.0 to 344.4 ± 74.0 mL H₂·h^?1·l^?1 (AFBR-M) when HRT decreased from 8 to 1 h. Maximum H₂ yields for AFBR-T and AFBR-M were 1.93 ± 0.21 and 2.68 ± 0.48 mol H₂·mol^?1 sucrose, respectively. The main metabolites were acetic acid (31.3%–41.5%) and butyric acid (10.2%–20.7%) (AFBR-M) and acetate (20.1%–39.3%) and ethanol (14.3%–29.9%) (AFBR-T). Denaturing gradient gel electrophoresis profiles revealed selective enrichment of microbial populations responsible for H₂ production by the aceto-butyric route (AFBR-M) and ethanol-type fermentation (AFBR-T).     

变性梯度凝胶电泳技术及其在食品微生物研究中的应用

食品中的很多微生物无法分离培养,传统的微生物研究方法不仅费时费力,而且会造成偏差。变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)是一种不依赖于纯培养的分子技术,因其快速、灵敏、分析全面等特点已被广泛应用于微生物生态学的研究中。本文介绍了DGGE 的发展历程、原理、操作步骤及其在食品微生物研究中的进展,评价了该技术的优势和局限性,展望了其在食品领域内的应用前景。     

PCR-DGGE分析吉林市传统发酵豆酱中细菌的多样性

该研究以吉林市农家传统发酵豆酱为试材,利用PCR-DGGE技术分析豆酱不同发酵阶段细菌的生物多样性及变化规律。结果表明,豆酱发酵过程中细菌菌群种类较为丰富,主要有戊糖片球菌（Pediococcus pentosaceu）、乳明串珠菌（Leuconostoc lactis）、葡萄球菌属（Staphylococcus sp.）、屎肠球菌（Enterococcus faecium）、解淀粉芽孢杆菌（Bacillus amyloliquefaciens）、弗氏柠檬酸杆菌（Citrobacter freundii）、霍氏肠杆菌（Enterobacter hormaechei）、产酸克雷伯氏菌（Klebsiella oxytoca）、不可培养杆菌（uncultured bacterium）。其中乳酸菌是发酵过程中的优势菌,对豆酱的风味起着主要作用。传统豆酱发酵过程中菌群有一定的亲缘性,菌落结构变化较小,为传统发酵豆酱的进一步开发和研究提供理论基础。     

Diversity analysis and quantification of lactic acid bacteria in traditionally fermented yaks’ milk products from Tibet

Fermented yaks’ milk is a characteristic fermented dairy product with a rich microflora (especially lactic acid bacteria [LAB]), that has been produced by Tibetan people in China throughout history. Systematic analysis of the LAB composition of fermented yaks’ milk from central Tibet is essential. In the present study, 38 samples of fermented yaks’ milk were collected from three regions in central Tibet. From these, a total of 211 isolates of LAB were generated using traditional pure culture methods. To accurately analyze their biodiversity, 16S rRNA gene sequence analysis and denaturing gradient gel electrophoresis (DGGE) were used. The isolates were identified as belonging to six genera and 22 species/subspecies. Among them, Lactobacillus species predominated (117 isolates) accounting for 55.45% of all isolates. Overall, L. delbrueckii subsp. bulgaricus (60 isolates; 28.43% of the total) was the most commonly isolated species of LAB from fermented yaks’ milk from central Tibet. Seven Lactobacillus species and the Bifidobacterium genus were confirmed by quantitative PCR (q-PCR). The q-PCR results showed that the abundance of these groups in fermented yaks’ milk from central Tibet ranged from 1.12 ± 0.36 log cfu/ml (for L. sakei from Ningzhong) to 7.77 ± 0.84 log cfu/ml (for L. helvetivus from Ningzhong). Lactobacillus acidophilus, L. delbrueckii subsp. bulgaricus, L. fermentum, L. helveticus varied in concentration depending on the region. The results indicated that traditional fermented yaks’ milk from different regions of central Tibet have complex compositions of LAB species. The diversity of LAB might be related to geographical origin.     

The bacterial community structure during bioleaching of a low-grade nickel sulphide ore in stirred-tank reactors at different combinations of temperature and pH

The focus of this study is to characterize the bacterial community structure present during stirred-tank bioleaching of a low-grade nickel sulphide ore at different temperatures (5 to 45 °C) and pH levels (3 and 5). This is a continuation of previous work, which was designed to assess the technical feasibility of applying elevated-pH bioleaching to this same ore from Manitoba, Canada. A combination of classical microbiological and molecular biological techniques has been used to identify and enumerate the members of the bacterial consortia over the course of the five-week experiments. DGGE analysis revealed the presence of at least 16 distinct 16S rRNA gene sequences, 14 of which are closely related to existing GenBank sequences. Two sequences were detected that are not closely related to existing GenBank sequences and may possibly be from novel species. Thirteen sequences are related to gene sequences of genera that have previously been detected in bioleaching environments (Acidithiobacillus, Leptospirillum, Sulfobacillus, Acidiphilium, Ferrimicrobium, and Acidimicrobium). Members from the genus Acidithiobacillus were dominant at all temperatures except 45 °C, at which Sulfobacillus spp. were dominant. Many of the acidithiobacilli are most closely related to strains of Acidithiobacillus ferrooxidans, and different strains were dominant under different experimental conditions, indicating considerable phenotypic heterogeneity within the species.     

Study of the Lactobacillus sakei protective effect towards spoilage bacteria in vacuum packed cooked ham analyzed by PCR-DGGE

The effectiveness of Lactobacillus sakei B-2 inoculated as a protective culture on the inhibition of spoilage bacteria on sliced vacuum packed cooked ham was investigated by using culture-dependent and -independent approaches. Total microbial DNA was directly extracted from both control and treatment samples, and subjected to a nested PCR protocol, PCR–DGGE analysis was used to identify and monitor the dynamic changes in the microbial population, followed by partial 16S rDNA sequencing. The DGGE profile demonstrated that the protective culture effectively suppressed growth of predominant spoilage bacteria L. sakei, Lactobacillus curvatus and Leuconostoc mesenteroides in cooked ham during storage at 4 °C, however, growth of uncultured Leuconostoc was not inhibited. The shelf-life of this product inoculated with L. sakei B-2, at levels of 5.91 ± 0.04 log₁₀ CFU g⁻¹ was 35 days, compared to 15 days of control samples, when the ham was stored at 4 °C.     

Microbiological characterisation of Robiola di Roccaverano cheese using PCR-DGGE

The aim of this study was the microbiological characterisation of a typical Italian cheese “Robiola di Roccaverano” with Protected Designation of Origin (PDO). Fresh and ripened robiola samples were collected from four artisanal and one industrial producer. Artisanal producers used raw goat's milk and natural fermentation, whilst the industrial producer used mixed cow–goat's milk and selected starters. The microbial communities were monitored during different seasons and ripening times with PCR–DGGE and culture-dependent methods.The cluster analysis showed that the DGGE bacterial patterns were related to the different manufacturing and climatic conditions, revealing the occurrence of species associated to Robiola di Roccaverano. The DGGE profiles of yeasts were affected by ripening in summer season.Moreover, the results obtained allowed the identification of microbial species such as Lactococcus lactis subsp. lactis, Geotricum spp. and Kluyveromyces lactis that are related to the production of this typical cheese.     

PLFA法和DGGE法分析制曲过程微生物群落变化

同时采用PLFA(Phospholipid fatty acid)谱图分析法和DGGE(Denaturing gradient gel electrophoresis)法分析大曲制曲过程中微生物群落的变化.结果表明:真菌类微生物生物量呈现先下降后上升趋势,发酵第7天各种微生物含量达到极限值,随后呈下降的趋势.通过贮存期...