1种食用菌细菌隐性污染栽培种的菌种分离研究

研究1种以木屑为主要配方的食用菌细菌隐性污染栽培种的菌种分离。稀释涂布平板法分析细菌总数。栽培种捣碎后悬于无菌水,分别以短时(5 min)涡漩振荡、摇床振荡(180 r/min)、低温静置(4℃) 3种方式处理;灭菌纱布过滤收集、无菌水淋洗木屑接种至PDA斜面,培养使其生长。以未经处理木屑为接种物作为对照。结果显示栽培种细菌数为15500 cfu/g;对照（30支）以及采用短时涡漩振荡处理所接30支斜面全部细菌污染;摇床振荡1、2、3、4天纯化率分别为20%、30%、46.67%及63.3%;低温静置1、2、3、4天纯化率分别为3.3%、10%、46.67%和60%。以木屑为主要配方的细菌隐性污染栽培种悬于无菌水经1~4天振荡或浸湿再分离的基内菌丝分离法可有效分离纯化菌种。     

牧区奶干渣组合式发酵剂响应面法优化

以脱脂牦牛乳为试验材料,发酵3.5 h滴定酸度为试验指标,在不同菌种最佳组合方式筛选的研究基础上,利用响应面Box-Behnken中心组合试验研究了4株筛自牧区、发酵性能优良的菌种:保加利亚乳杆菌(MGD1-3)、嗜热链球菌(MGB39-5)、嗜热链球菌(G81-1)、植物乳杆菌(BM5152)不同接种比例对牧区奶干渣发酵效果的影响。结果表明,优化得到最优组合式发酵剂为:MGD1-3体积分数3.06%、MGB39-5体积分数0.64%、G81-1体积分数0.55%、BM5152体积分数2.04%,滴定酸度为75.87°T。为了便于实际操作,确定最终体积接种比为:MGD1-3∶MGB39-5∶G81-1∶BM5152=30∶6∶5∶20,接种量体积分数为6.0%,该条件下进行3次平行试验,得到滴定酸度平均值为74.21°T;将接种优化后组合式发酵剂制成的奶干渣(试验组)与牧区采集奶干渣(对照组)进行感官与理化指标比较,结果表明该组合式发酵剂发酵性能优良。     

畜禽场排出空气的净化技术

详细介绍了当前国内外对畜禽场排出空气的主要净化技术和性能,对技术的设计原理、材料选择与净化效果进行了分析评价。     

高效脱色菌的特性及其在染化废水厌氧处理中的生物强化作用

从活性污泥等微生物源中分离、筛选获得10株高效脱色菌，经鉴定其属于Pseudomonas,Bacillus，Xanthomonas，Erwinia，Alealigenes，Plesiomonas。对艳红染料生产废水脱色试验表明最适温度30℃，pH6.6～8.0，湿细胞浓度20～30!g*L-1;静置培养（兼氧）脱色率高于振荡培养；外加NADH、NADPH、易降解有机物和有机废水可强化脱色菌的脱色性能。为实现脱色菌的资源化应用，试验将PseudomonasD18,ErwiniaD17,AlcaligenesD19andPlesiomonasD124株脱色菌制成混合培养物和固定化细胞，分别投加于处理染化废水的厌氧反应器，结果发现在相同COD负荷、水力负荷条件下，投加固定化细胞的反应器，其脱色率可提高5!%～10!%，出水苯胺浓度提高40!%～65!%。电镜观察发现脱色菌在胶团内外持留、增殖。厌氧反应器性能的改善主要通过功能性微生物生物量和相关基因库量的增加而实现。     

不同土壤中硅酸盐细菌生理生化特征及其解钾活性的研究

在以钾长石粉为唯一 K 源的硅酸盐细菌选择性培养基上,从我国部分省市土壤中筛选到 16 株硅酸盐细菌,以本实验室保藏 NBT 菌株为参照,对其生理生化特性、耐盐性、抗生素抗性、温度敏感性及释K 能力等生物学特性进行了测定。结果表明,17 株硅酸盐细菌菌体均为杆状,产生椭圆至圆形芽胞。其中 SB6、SB13 为短杆状,SB4、SB6、SB10、SB15 菌株是 G ,其余菌株是 G-。NH4 、NO3 为良好 N 源,且能在无 N -培养基上生长。菌株 SB13 和 NBT 解 K 能力较强,释放的 K 比接灭活菌对照分别增加 49.1 %和 45.3 %。菌株SB2、SB4、SB5、SB6 在 20 g/L NaCl 浓度的培养基上能生长,在温度为 10 ~ 40 范围内供试硅酸盐细菌能够良好生长。     

7株解有机磷细菌的分离和鉴定

从土壤中分离筛选出7株解有机磷的微生物。对这7株解磷细菌进行了形态、生理生化性状测定及16SrDNA序列分析(GenBankaccessionNo:S2,AY651922;S3,AY661923;X1,AY651925;Y1,AY651924;H1,AY663435;H2,AY663436andHe,AY663436)。其中S2、S3、X1和He属于假单胞菌属(Pseudomonas),Y1属于芽孢杆菌属(Bacillus),H1属于不动杆菌属(Acinetobacter),H2属于寡养单胞菌属(Stenotrophomonas)。进一步通过G C含量和DNA-DNA杂交研究,结果表明,S2、S3和X1为产碱假单胞菌(Pseudomonasalcaligenes),Y1为蜡状芽孢杆菌(Bacilluscereus)。     

ATP发光技术测定辐照前脱水蔬菜和调味品的含菌量

用ATP发光技术测定脱水蔬菜和调味品的初始含菌量,发现脱水蔬菜和调味品中细菌ATP的生物发光强度与其初始含菌量之间显著相关,从样品的制备到获得ATP发光强度结果的检测过程仅需1～2h。该技术可用于辐照灭菌前的微生物检测。     

活性光合细菌对改善水产养殖体系水质状况的作用初探

在鱼虾育苗、养殖，池塘淡水养鱼以及观赏鱼养殖中，使用以牛肉水提取液为主要培养基培养的活菌数大于50亿／mL的活性光合细菌，测定了使用前后水的pH值、COD值、NH3-N、亚硝酸盐、溶解氧等指标，结果表明：与市场出售的光合细菌(活菌数为25亿／mL)相比，活菌数大于50亿／mL活性光合细菌能明显改善水产养殖体系的水质状况。     

Survival of bacterial DNA and culturable bacteria in archived soils from the Rothamsted Broadbalk experiment

Dried soil samples from many sources have been stored in archives world-wide over the years, but there has been little research on their value for studying microbial populations. Samples collected since 1843 from the Broadbalk field experiment on crop nutrition at Rothamsted have been used to document changes in the structure and composition of soils as agricultural practices evolve, also offering an invaluable record of environmental changes from the pre- to post-industrial era in the UK. To date, the microbial communities of these soils have not been studied, in part due to the well-documented drop in bacterial culturability in dried soils. However, modern molecular methods based on PCR amplification of DNA extracted directly from soil do not require bacterial cells to be viable or intact and may allow investigations into the legacy of bacteria that were present at the time of sample collection.

In a preliminary study, to establish if dried soils can provide a historical record of bacterial communities, samples from the Broadbalk soil archive dating back to 1868 were investigated and plots treated with either farmyard manure (FYM) or inorganic fertilizer (NPK) were compared. As anticipated, the processes of air-drying and milling greatly reduced bacterial viability whilst DNA yields declined less and may be preserved by desiccation. A higher proportion of culturable bacteria survived the archiving process in the FYM soil, possibly protected by the increased soil organic matter. The majority of surviving bacteria were firmicutes, whether collected in 2003 or in 1914, but a wide range of genera was detected in DNA extracted from the samples using PCR and DGGE of 16S rRNA genes. Analysis of DGGE band profiles indicated that the two plots maintained divergent populations. Sequence analysis of bands excised from DGGE gels, from a sample collected in 1914, revealed DNA from - and β-proteobacteria as well as firmicutes. PCR using primers specific for ammonia oxidizing bacteria showed similar band profiles across the two treatments in recently collected samples, however older samples from the NPK plot showed greater divergence. Primers specific for the genus Pseudomonas were designed and used in real-time quantitative PCR to indicate that archived soil collected in 1868 contained 10-fold less pseudomonad DNA than fresh soil, representing around 10⁵ genomes g⁻¹ soil. Prior to milling, dramatically less pseudomonad DNA was extracted from recently collected air-dried soil from the NPK compared to the FYM plot; otherwise, the two plots followed similar trends. Overall bacterial abundance, diversity and survival during the archiving process differed in the two soils, possibly due to differences in clay and soil organic matter content. Nevertheless, the results demonstrate that air-dried soils can protect microbial DNA for more than 150 years and offer an invaluable resource for future research.     

高产水稻土细菌多样性的培养法与非培养法比较研究

利用细菌的通用引物扩增江西余江县高产水稻土红壤细菌总DNA和平板培养细菌混合总DNA的16S rDNA基因片段,在此基础上分别建立两种16S rDNA文库(文库a和文库b)。从两个文库中各随机挑选100个克隆,扩增出阳性克隆中的插入片段后选用HhaⅠ和RsaⅠ两种四碱基酶进行ARDRA(amplified rDNA restriction analysis)分析。统计比较分析发现,文库a的Shannon-Wienner指数、Simpson指数、丰富度、均一度分别为4.432、0.987、18.885和0.973,均高于文库b中相应的多样性参数(分别为2.271、0.758、5.736和0.501),即平板培养方法所展现的细菌群落结构多样性低于土壤中原始的多样性。结果表明,传统培养方法存在着很大的局限性,必须结合新的分子生物学技术手段才能更全面完善地认识土壤微生物群落结构多样性,以期充分利用其中丰富的微生物资源。