我国典型湿热地区变质烟用香精中腐败微生物的多样性分析

【背景】烟用香精是卷烟生产中的重要辅料,有些香精,特别是剩余的加料香精在运输和储存等过程中可能污染微生物,引起腐败变质发生。【目的】分析变质烟用香精中腐败微生物的群落多样性,确定引起加料香精变质的主要腐败微生物。【方法】采用稀释平板法,对我国典型湿热地区云南、福建和广西三地变质加料香精样品中的污染细菌、霉菌或酵母进行分离,分析其16S rRNA、ITS或26S rRNA基因序列,并通过反证试验确定引起变质的主要腐败微生物。【结果】所考察3个典型湿热地区变质加料香精存在不同程度的细菌、霉菌或酵母污染,其中细菌污染最为普遍。分离的76株污染细菌分布在2门4纲5目10科15属,细菌的群落多样性比较丰富,其中雷尔氏菌属(Ralstonia)、柠檬酸杆菌属(Citrobacter)、肠杆菌属(Enterobacter)及芽孢杆菌属(Bacillus)是优势菌群;9株霉菌分布在子囊菌门(Ascomycota)的3纲6属,每个属分离到1-2株霉菌;42株酵母分布在子囊菌门的6个属,其中毕赤酵母属(Pichia)、细枝钩酵母菌属(Wickerhamomyces)及酵母菌属(Saccharomyces)是优势菌群。云南和福建两地样品的细菌菌群组成明显多于广西,广西样品的霉菌菌群组成以及福建样品的酵母菌群组成均分别多于其余两地。反证试验结果显示,柠檬酸杆菌属、肠杆菌属、芽孢杆菌属、雷尔氏菌属和毕赤酵母属是引起烟用加料香精变质的主要腐败微生物。【结论】变质加料香精中污染微生物菌群组成及腐败微生物确定的研究将有助于我国湿热地区香精香料中污染微生物的控制和腐败变质的预防。     

石油降解菌的分离鉴定及石油污染土壤的细菌多样性

从石油污染的土壤中分离筛选到28株石油降解菌,经鉴定分别为短杆菌属、假单胞菌属、邻单胞菌属和微球菌属;对4个石油不同程度污染的土壤样品中嗜油微生物分布状况进行分析,发现污染严重的土壤样品中嗜油菌的数量相对较多;用聚合酶链式反应(PCR)、变性梯度凝胶电泳(DGGE)和切胶测序相结合的方法对4个土壤样品中的细菌多样性进行分析,结果显示在受污染的土壤中,My cobacterium和B acillus在污染程度较低的样品中分布的较为集中,F lavobacterium和A zosp ira在污染程度较高的样品中丰度较高。属于B eta p roteobacterium类群的细菌在受污染的土壤中占有优势,同时还有一些不可培养的菌群存在。气质联用(GC-M S)分析结果表明石油污染程度及污染物中芳香烃类的含量对细菌多样性有着显著影响。在石油污染程度高,芳香烃类含量高的样品中细菌的多样性相对较低。     

氧化塘中细菌种群组成动态

对武汉地区氧化塘中的细菌数量、细菌种类、种群组成、优势种及群落多样性进行了初步研究。从氧化塘中分离的细菌经鉴定属于12个属,主要有假单胞菌属(Pseudomonas)、棒杆菌属(corynebacterium)、无色杆菌属(Achromobacter)和微杆菌属(Microbacterium),其它菌属是芽孢杆菌属(Bacillus)、色杆菌属(Chromobacterium)、短杆菌属(Brevibacterium)、葡萄球菌属(Staphylococcus)、微球菌属(Micrococcus)、黄杆菌属(Flavobacterium)、埃希氏菌属(Escherichia)和链球菌属(Streptococcus)。在5个小塘中,细菌数量、种群组成和群落多样性指数都有不同,这与各塘的污染程度有关。由此,多样性指数可用于监测氧化塘的净化效果。     

养殖大黄鱼细菌菌群分离鉴定与分析*

对新鲜大黄鱼感官、化学、微生物品质和细菌菌群组成进行定性和定量分析。结果表明,细菌总数为5．51±0．25log10cfu／g,挥发性盐基氮为7．84±2．25mg／100g。分离获得279株细菌,84．2％是革兰氏阴性菌,少量革兰氏阳性菌被检出(6．1％)。优势菌群是肠杆菌科(14．7％)、气单胞菌属(12．5％)、不动杆菌属(11．5％)、摩氏杆菌属(11．1％),并出现了一定比例的假单胞菌属、嗜麦芽窄食单胞菌和其他细菌。肠杆菌科出现较多,表明大黄鱼细菌污染主要来自养殖海区,受非原有菌污染较重,应引起重视。     

养殖大黄鱼细菌茵群分离鉴定与分析

对新鲜大黄鱼感官、化学、微生物品质和细菌菌群组成进行定性和定量分析。结果表明,细菌总数为5．51±0．25log10cfu／g,挥发性盐基氮为7．84±2．25mg／100g。分离获得279株细菌,84．2％是革兰氏阴性菌,少量革兰氏阳性菌被检出(6．1％)。优势菌群是肠杆菌科(14．7％)、气单胞菌属(12．5％)、不动杆菌属(11．5％)、摩氏杆菌属(11．1％),并出现了一定比例的假单胞菌属、嗜麦芽窄食单胞菌和其他细菌。肠杆菌科出现较多,表明大黄鱼细菌污染主要来自养殖海区,受非原有菌污染较重,应引起重视。     

两种不同生境中国沙棘种子内生细菌的多样性

植物内生菌广泛分布于植物的各种组织及器官中,对植物的生长表现出各种作用,而植物种子中的内生菌对植物的作用也越来越受到人们的关注。以榆中县和秦安县两种不同生境的中国沙棘种子为材料,分析中国沙棘种子内生细菌多样性,以期探究生境对种子内生菌多样性的影响,并为进一步研究种子内生菌与沙棘的相互作用提供依据。研究利用纯培养方法和高通量测序方法分别进行中国沙棘种子内生细菌多样性分析。对纯培养分离得到的内生细菌,利用16S rRNA基因序列分析法结合形态学特征进行内生细菌的鉴定;对高通量测序得到的数据进行基于OTUs(Operational Taxonomic units,可操作分类单元)的物种注释分析。通过纯培养方法从榆中县中国沙棘种子中分离得到4株内生细菌,分属于芽孢杆菌属(Bacillus)、葡萄球菌属(Staphylococcus)和假单胞菌属(Pseudomonas);秦安县中国沙棘种子中分离得到5株内生细菌,分属于芽孢杆菌属(Bacillus)、葡萄球菌属(Staphylococcus)和不动杆菌属(Acinetobacter)。采用高通量测序方法检测到榆中县中国沙棘种子内生细菌分属于7个门、68个属,秦安县中国沙棘种子内生细菌分属于5个门、30个属。在门分类水平,榆中县中国沙棘种子内生细菌的主要优势门类是蓝藻门(Cyanobacteria)和变形菌门(Proteobacteria),相对丰度分别为95.62%和2.03%;秦安县中国沙棘种子的主要优势门类是变形菌门(Proteobacteria)和蓝藻门(Cyanobacteria),相对丰度分别为91.68%和8.06%。在属分类水平,榆中县中国沙棘种子内生细菌的优势菌属为蓝藻细菌属(Cyanobacteria),相对丰度为95.09%;秦安县中国沙棘种子内生细菌的优势菌属为寡养单胞菌属(Stenotrophomonas),相对丰度为85.60%。榆中县和秦安县两种不同生境中国沙棘种子内生细菌有着丰富的多样性,且内生细菌的多样性和丰富度存在明显差异,表现为榆中县中国沙棘种子内生细菌的多样性和丰富度均高于秦安县中国沙棘种子内生细菌。     

微繁殖系统中出现的细菌污染

植物组织培养研究者和微繁殖者常常通过肉眼观察来确定培养物是否有污染物.但是,Neo Plants Ltd公司和诺丁汉大学农业学校的C.Leifert及其同事发现,大多数被污染的植物组织培养物看起来都是健康的,尽管其繁殖速度减慢.Leifert等研究了从落新妇、假升麻、墨西哥桔、黄栌、翠雀、扶郎花、黄花菜、玉簪、鸢尾、报春花和唐松草等属植物上分离的198种细菌污染物,发现90%属于芽孢杆菌属、肠杆菌属、微球菌属、葡萄球菌属和乳杆菌属.通过比较从新鲜外植体和培养了一年多的植株上分离的微生物以及考虑污染物的自然生境,可以发现污染的来源. Leifert等进行的研究表明,1/3～1/2的污染是由于操作引起的1/4～1/2的污     

北极海洋沉积物中锰细菌的分离与系统发育

对中国第二次北极科学考察采集的北极海洋沉积物中的锰细菌进行了筛选、分离和系统发育分析。根据其在筛选平板上菌落的形态学特征,分别从站位P11和S11采集的沉积物中分离到了21株和19株锰细菌。系统发育分析表明,两个站位的锰细菌群落组成有着明显的差别。站位P11分离的可培养锰细菌主要由细菌域（Bacteria）中变形杆菌门的γ-变形杆菌纲（γ-Proteobacteria）和放线菌纲（Actinobacteria）组成,二者分别占86％和14％;γ-变形杆菌纲主要包括嗜冷杆菌属（Psychrobacter）、希瓦氏菌属（Shewanella）、假交替单胞菌属（Pseudoaheromonas）、不动杆菌属（Acinetobacter）、海杆菌属（Marinobacter）,其中以嗜冷杆菌属为主,其比例可达67％。从站位S11分离到的可培养锰细菌主要包括细菌域中变形杆菌门的α-变形杆菌纲（α-Proteobacteria）和γ-变形杆菌纲以及拟杆菌门（Bacteroides）中的黄杆菌纲（Flavobaeteria）;γ-变形杆菌纲主要包括希瓦氏菌属、海单胞菌属（Marinomonas）和交替单胞菌属（Aheromonas）,α-变形杆菌纲主要为鞘氨醇单胞菌属（Sphingomonas）。实验菌株均对Mn^2＋有着较强的抗性,其中以菌株Marinomonas sp．S11-S-4耐受性最高。     

处理石油化工废水的活性污泥中优势微生物群系及其降解效能的研究

处理石油化工废水的活性污泥中微生物以细菌为主体,霉菌、酵母菌数量较少。分离到167株细菌,主要群系为不动细菌属,假单胞菌属,产碱杆菌属、微球菌属、棒状杆菌属、芽孢杆菌属等、其中大部分菌株对苯酚、苯乙烯、丙酮、甲醇具有降解效能,尤其对苯酚、苯乙烯降解能力较强,菌株数量也较多。     

对叶榕榕果内生细菌分离鉴定及抑菌活性的研究

【目的】为植物-内生细菌的生态关系研究, 以及植物内生细菌资源的利用提供一定的依据。【方法】采用微生物学传统分离培养的方法从对叶榕果实中分离到内生细菌54株, 通过限制性酶切分析(ARDRA)共有16个操作分类单元(Operational taxonomic units, OTUs), 对16株代表菌株16S rDNA序列系统发育分析。【结果】16个菌株都找到了与其相似性最高的菌株, 相似性达到95%?100%。其中6株为芽孢杆菌Bacillus属, 为对叶榕果实内生细菌优势菌属; 3株为Staphylococcus属, 2株为Pseudomonas属, 1株为Serrata属, 1株为非培养细菌的同源菌, 1株为Kocuria属, 1株为Delftia属, 还有1株为Acinetobacter属。【结论】这16株内生菌在系统发育树中明显聚为两大支; 在参与抑菌试验的14株内生菌中, 有13株对受试菌有不同程度的拮抗作用。尤其是其中的芽孢菌属的Swx15和不动杆菌属的Swx25菌株, 抑菌作用较强, 且有较广的抑菌谱性。     

Genetic diversity and involvement in bread spoilage of Bacillus strains isolated from flour and ropy bread

AIMS: To study Bacillus contamination of wheat flour and ropy bread, to analyse genetic diversity of isolated strains and to evaluate the ability of these strains to produce ropy bread. METHODS AND RESULTS: Classical and molecular methods [16S rDNA sequencing and random amplified polymorphic DNA (RAPD)-PCR] were used to identify and type-isolated strains. The predominant species isolated were Bacillus subtilis and B. licheniformis. RAPD analysis demonstrated that the same sample may harbor different strains. Ten of 15 strains of B. subtilis and four of six strains of B. licheniformis were able to cause rope spoilage of the laboratory-baked bread. CONCLUSION: RAPD typing can be useful in the tracking of Bacillus strains during bakery processing and in the understanding of the role of different Bacillus strains in the rope spoilage of bread. SIGNIFICANCE AND IMPACT OF THE STUDY: The results indicate the variability of Bacillus strains isolated from flour and responsible for rope spoilage of bread.     

Survival and growth of Bacillus cereus in bread

Bread doughs were artificially inoculated with spores of six Bacillus cereus strains at different inoculum levels and counts of survivors in bread determined during storage at 27.5 degrees C. No B. cereus were isolated from the centre crumb of 400 g loaves when the dough contained less than 10(4) spores/g whereas with 800 g loaves survival occurred with doughs containing 5.0 X 10(3) spores/g. With all strains there was a period of at least 24 h before multiplication took place in the bread. The inclusion in dough of 0.2% of calcium propionate, based on flour, effectively delayed germination and subsequent multiplication of B. cereus spores. It is concluded that the risk of food poisoning due to the presence of B. cereus in bread is minimal.     

Survival and growth of Bacillus cereus in bread

Bread doughs were artificially inoculated with spores of six Bacillus cereus strains at different inoculum levels and counts of survivors in bread determined during storage at 27·5°C. No B. cereus were isolated from the centre crumb of 400 g loaves when the dough contained less than 10⁴ spores/g whereas with 800 g loaves survival occurred with doughs containing 5·0 times 10³ spores/g. With all strains there was a period of at least 24 h before multiplication took place in the bread. The inclusion in dough of 0·2% of calcium propionate, based on flour, effectively delayed germination and subsequent multiplication of B. cereus spores. It is concluded that the risk of food poisoning due to the presence of B. cereus in bread is minimal.     

Rope-producing strains of Bacillus spp. from wheat bread and strategy for their control by lactic acid bacteria

Two types of white wheat bread (high- and low-type loaves) were investigated for rope spoilage. Thirty of the 56 breads tested developed rope spoilage within 5 days; the high-type loaves were affected by rope spoilage more than the low-type loaves. Sixty-one Bacillus strains were isolated from ropy breads and were characterized on the basis of their phenotypic and genotypic traits. All of the isolates were identified as Bacillus subtilis by biochemical tests, but molecular assays (randomly amplified polymorphic DNA PCR assay, denaturing gradient gel electrophoresis analysis, and sequencing of the V3 region of 16S ribosomal DNA) revealed greater Bacillus species variety in ropy breads. In fact, besides strains of B. subtilis, Bacillus licheniformis, Bacillus cereus, and isolates of Bacillus clausii and Bacillus firmus were also identified. All of the ropy Bacillus isolates exhibited amylase activity, whereas only 32.4% of these isolates were able to produce ropiness in bread slices after treatment at 96 degrees C for 10 min. Strains of lactic acid bacteria previously isolated from sourdough were first selected for antirope activity on bread slices and then used as starters for bread-making experiments. Prevention of growth of approximately 10(4) rope-producing B. subtilis G1 spores per cm(2) on bread slices for more than 15 days was observed when heat-treated cultures of Lactobacillus plantarum E5 and Leuconostoc mesenteroides A27 were added. Growth of B. subtilis G1 occurred after 7 days in breads started with Saccharomyces cerevisiae T22, L. plantarum E5, and L. mesenteroides A27.     

Treatment of bran containing bread by baking enzymes; effect on the growth of probiotic bacteria on soluble dietary fiber extract in vitro

Different ways of treating bran by baking enzymes prior to dough making and the baking process were used to increase the amount of water-soluble dietary fiber (DF) in wheat bread with added bran. Soluble DF was extracted from the bread with water and separated from the digestible material with gastrointestinal tract enzymes and by solvent precipitation. The baking enzyme mixtures tested (xylanase and glucanase/cellulase, with and without lipase) increased the amounts of soluble arabinoxylan and protein resistant to digestion. The isolated fiber was used as a growth substrate for 11 probiotic and intestinal Bifidobacterium strains, for commensal strains of Bacteroides fragilis and Escherichia coli, and for potential intestinal pathogenic strains of E. coli O157:H7, Salmonella typhimurium, and Clostridium perfringens. Fermentation analyses indicated that the tested strains had varying capacity to grow in the presence of the extracted fiber. Of the tested probiotic strains B. longum species generally showed the highest ability to utilize the fiber extracts, although the potential pathogens tested also showed an ability to grow on these fiber extracts. In sum, the enzymes used to improve the baking process for high-fiber bread can also be used to produce in situ soluble fiber material, which in turn can exert prebiotic effects on certain potentially beneficial microbes.     

In vitro growth and acid production by mutans streptococci on traditional African foods

The growth rate and production of acids by mutans streptococci (MS) are influenced by their ability to ferment different dietary carbohydrates. This suggests that the nutrient environment in the oral cavity affects bacterial virulence. The aim of this study was to investigate the effect of maize, samp and brown bread on the growth and acidogenicity of this species. Six laboratory references and five clinical strains isolated from the dental plaque of South African black and 'colored' (historical race classification) children were studied in batch culture on maize, samp (coarsely ground maize), brown bread and compared against a 3% sucrose control. The doubling time of bacterial strains was prolonged in maize (1.9-17.5 h) and samp (2.4-18.4 h), and the number of cell divisions was low. Staple foods accounted for 25% (F=5.98; P=0.0007) and MS strains 30.78% (F=2.84; P=0.009) of the total variance. The fermentation of samp and maize showed the least drop in pH of the culture medium, ranging between 0.54 and 1.06 and 0.69 and 2.28 pH units respectively, with variation between strains most significant in maize (F=33.62; P<0.0001). The total mean concentration of acids produced was highest in bread (25.13 mM/mL) and samp (17.00 mM/mL) which was comparable to Brain Heart Infusion broth (16.49 mM/mL) and a basal synthetic medium (17.96 mM/mL) containing 3% sucrose, but the yield of lactate, acetate and formate was low during the fermentation of samp (0.50 mM/mL), BHI+3% sucrose (4.12 mM/mL) and brown bread (0.06 mM/mL) respectively. Results indicated that maize and samp do not optimally support the growth or acid production by MS, and the varying response of test strains demonstrates the strain variability of this species to different carbohydrate sources in the diet.     

Phytase-active yeasts from grain-based food and beer

Aims: To screen yeast strains isolated from grain‐based food and beer for phytase activity to identify high phytase‐active strains. Methods and Results: The screening of phytase‐positive strains was carried out at conditions optimal for leavening of bread dough (pH 5·5 and 30°C), in order to identify strains that could be used for the baking industry. Two growth‐based tests were used for the initial testing of phytase‐active strains. Tested strains belonged to six species: Saccharomyces cerevisiae, Saccharomyces pastorianus, Saccharomyces bayanus, Kazachstania exigua (former name Saccharomyces exiguus), Candida krusei (teleomorph Issachenkia orientalis) and Arxula adeninivorans. On the basis of initial testing results, 14 strains were selected for the further determination of extracellular and intracellular (cytoplasmic and/or cell‐wall bound) phytase activities. The most prominent strains for extracellular phytase production were found to be S. pastorianus KVL008 (a lager beer strain), followed by S. cerevisiae KVL015 (an ale beer strain) and C. krusei P2 (isolated from sorghum beer). Intracellular phytase activities were relatively low in all tested strains. Conclusions: Herein, for the first time, beer‐related strains of S. pastorianus and S. cerevisiae are reported as phytase‐positive strains. Significance and Impact of the Study: The high level of extracellular phytase activity by the strains mentioned previously suggests them to be strains for the production of wholemeal bread with high content of bioavailable minerals.     

Application of Bifidobacterium strains to the breadmaking process

The possible use of bifidobacterial strains from different origin (adult and infant humans, and chicken) as novel starter cultures for breadmaking was evaluated. Fermentative parameters of doughs (pH, volume, total titrable acidity [TTA], lactic and acetic acids production and rheofermentative parameters) and technological parameters of breads (specific volume, bread shape and crumb hardness) were analyzed. Human bifidobacterial strains could replace Lactobacillus strains, commercialized for breadmaking, as they yielded breads with similar characteristics but with the advantage of having softer crumbs. Important differences between the behavior of chicken bifidobacterial strains and human bifidobacterial strains were found when comparing bread TTA, bread shape and bread volume. Breads made with chicken strains showed significantly lower (p < 0.05) specific bread volume than those made with human strains, while showing similar values of TTA. The effects observed when using bifidobacterial strains from different origin as novel starter cultures for breadmaking seemed to depend on the strain and its origin.     

Characterization of the yeast population from traditional corn and rye bread doughs

M.J. ALMEIDA AND C.S. PAIS. 1996. Yeasts were isolated from a variety of home-made bread doughs and identified. A pure culture of Saccharomyces cerevisiae was found in 18% of the doughs. The same species predominated in 80% of the doughs examined whereas Issatchenkia orientalis, Pichia membranaefaciens and Torulaspora delbrueckii were present in about 40% of the samples. About one quarter of the isolates displayed killer activity, strains of P. anomala showing the broadest spectra. Two isolates of S. cerevisiae and three of T. delbrueckii gave biomass values in sucrose medium similar to or higher than those obtained with commercial compressed baker's yeast strains.     

Differentiation ofLactobacillus spp. isolated from Chinese sourdoughs by AFLP and classical methods

TwentyLactobacillus strains isolated from sourdoughs that were used as starters for Chinese steamed bread were characterised by amplified fragment length polymorphism (AFLP) and classical methods. Twenty strains were divided into five clusters with AFLP profiles using similarity coefficient 0.60 (Jaccard coefficient) as a cut-off point, while three bio-clusters were formed with a numerical taxonomy classification based on phenotypic features, at a similarity level of 0.80 (Jaccard coefficient) and an average linkage algorithm (UPGMA) of clustering. A good correlation was observed between the two methods. The results suggested that the AFLP method is a useful taxonomic tool for typing of LAB, and the combination of the two methods may lead to obtain an improved understanding of the relationship between the phenotype and genotype of theLactobacillus spp. from the sourdough.