短芒大麦草青贮微生物特性研究及优良乳酸菌筛选

以短芒大麦草为研究对象,利用传统培养法从叶围和青贮发酵体系中分离出乳酸菌、大肠杆菌、好氧细菌、酵母菌和霉菌,并计数;结合细菌形态学、生理生化特征及16S rDNA序列分析鉴定分离出的乳酸菌菌株;通过研究乳酸菌的生长曲线、产酸特性及耐酸性,筛选优质乳酸菌。以期探明短芒大麦草叶围及青贮发酵体系中微生物菌群特性及青贮料中乳酸菌多样性,筛选出具有促发酵效果的乳酸菌菌株,为有益微生物饲料研发奠定基础。试验结果表明,短芒大麦草经青贮发酵后各微生物菌群数量发生不同程度变化,乳酸菌数量由0 cfu/g FM增加到4.00×10⁸ cfu/g FM,酵母菌数量由8.50×10⁵ cfu/g FM增加到1.02×10⁸ cfu/g FM,而好氧细菌、大肠杆菌和霉菌数量变化不明显;从短芒大麦草青贮发酵体系分离得到4株乳酸菌,经鉴定Lx36为Lactobacillus pentosus,Lx37为Lactobacillus brevis,Lx53为Pediococcus pentosaceus,Lx54为Lactobacillus parabuchneri;筛选得到1株益于青贮的乳酸菌株Lx36,约在20 h后进入稳定生长期,OD值达到4.21,且发酵12 h的pH仅为4.08,并可以在pH=3.0环境条件下生长。综上所述,青贮发酵是体系中各种微生物相互作用的过程,微生物菌群的数量及变化直接影响青贮饲料发酵品质。短芒大麦草青贮饲料中乳酸菌种类较丰富,筛选得到的戊糖乳杆菌繁殖速度快、产酸能力强同时表现出了较强的耐酸性,具有潜在的生产应用价值,适宜用作促发酵的青贮添加剂菌种。     

重组基因工程乳酸菌的研究进展

乳酸菌作为人和动物体内的正常菌群,具有诸多益生功能.随着分子生物学研究的不断深入,重组基因工程乳酸菌研究取得了较快进展.在构建表达外源功能性基因的重组基因工程乳酸菌、表达保护性抗原蛋白质用以递呈黏膜免疫抗原的过程中,乳酸菌质粒载体起到了至关重要的作用.电转化技术的应用,简化了操作过程,推动了重组基因工程乳酸菌的研究与应用.     

微生态制剂在海参养殖中的应用

本文就微生态制剂在海参养殖中的应用及应用中应注意的问题作一综述。     

添加乳酸菌和纤维素酶对苜蓿青贮品质的影响

初花期收获的苜蓿(Medicago sativa)经过0、8、32 h的晾晒,添加乳酸菌和纤维素酶进行青贮。结果表明,苜蓿凋萎（干物质含量为38.45%）青贮可以使青贮料的氨态氮含量显著降低,并保存有更多的粗蛋白质,生成更多的乳酸。苜蓿青贮时添加乳酸菌和纤维素酶能明显改善苜蓿青贮料的发酵品质,即降低青贮料的氨态氮含量和保存更多的粗蛋白质以及生成更多的乳酸。晾晒8 h的苜蓿添加乳酸菌（106 cfu/g）和纤维素酶（0.05 g/kg）的青贮效果最好;而晾晒32 h的苜蓿直接青贮的效果最好。     

银杏内生细菌防治辣椒疫病研究

从健康银杏植株叶片中分离到108 株内生细菌,通过平板测试从中筛选获得35 株对辣椒疫霉菌Phytophthora capsici具有拮抗作用的菌株.温室盆栽试验证明,菌株 Zy44、Fy11、Hy7、Hy14 和 Zy25 对辣椒疫病防效显著,其中Zy44 和 Fy11 发酵液在辣椒疫霉菌接种 20 d 后防效分别为 62%和 52.33%;经形态观察、生理生化特征测试、16S rDNA序列和gyrB基因序列分析,鉴定这两个菌株为解淀粉芽胞杆菌Bacillus amyloliquefaciens;其发酵滤液及酸沉淀获得的脂肽类抑菌物质对盆栽辣椒苗疫病的防治试验结果表明,菌株 Zy44 的防效高于 Fy11,且脂肽类物质的防效高于发酵滤液;显微镜观察发现菌株 Zy44的脂肽类抑菌物质可导致辣椒疫霉菌菌丝体膨大, 分枝增多, 成簇状,并显著抑制孢子囊的形成.     

Soil sterilization effects on root growth and formation of rhizosheaths in wheat seedlings

Sterilized soils are frequently used in experiments related to soil biology. Soil sterilization is known to alter physicochemical characteristics of soil, plant growth and community structure of the newly developed bacterial population. However, little information exists regarding soil sterilization effects on belowground processes mediated through root–microbe–soil interactions, e.g., development of rhizosheaths which significantly promote the plant growth under stress environments. The present study was conducted to elucidate effects of soil sterilization on wheat root growth and formation of rhizosheaths in relation to chemical changes caused by soil sterilization and the proportion of expolysaccharide (EPS)-producers in bacterial population recolonizing the sterilized soils. Wheat plants were grown for two weeks under greenhouse conditions either in the unsterilized soil or in soils sterilized by autoclaving (121 °C, 1 h) or by gamma (γ)-irradiation (50 kGy). While soil sterilization had no effect on the release of macronutrients, both sterilization procedures significantly increased the electrical conductivity, water-soluble carbon and DTPA-extractable Mn. Seedlings grown in sterilized soils produced higher root biomass and the rhizosheath soil (RS) mass as compared to those grown in the unsterilized soil. Soil sterilization also increased the root length, surface area, volume and number of tips. In bulk soil, RS and on roots, the proportion of EPS-producers in the total bacterial population was higher in sterilized treatments than in the unsterilized. Amending the unsterilized soil with glucose-C increased the root biomass, whereas adding Mn II increased the RS mass. The results showed that soil sterilization by autoclaving or γ-irradiation increases the root growth and RS mass of wheat seedlings. The water-soluble C and DTPA-extractable Mn released upon sterilization, and the increased proportion of EPS-producers in the bacterial population recolonizing the sterilized soils were involved in the observed effects. The results may have implications in studies using autoclaved or γ-irradiated soils to investigate soil–plant–microbe interactions and signify the need to account for intrinsic stimulatory effects of soil sterilization.     

好氧反硝化细菌的筛选

采集江安河及府河淤泥样本,采用BTB培养基与N-(1-萘基)-乙二胺光度法筛选出20株具有反硝化能力的好氧菌株.选取其中5株反硝化能力较强的DM1、DM2、DM3、DM4和DM5菌株,进行NO3--N去除率测定,其48 h NO3--N去除率均达到了30％以上.其中DM1、DM2、DM3和DM5菌株氮去除率依次为43.9％、47.6％、47.9％和51.3％.对DM5菌株进行生长曲线测定,进行pH值和温度对反硝化速率影响测定,试验结果表明在pH 7.0～7.4,温度20～30℃时,DM5菌株反硝化效果较好.     

不同施肥方式对红壤蔬菜田氨氧化细菌和氨氧化古菌群落的影响

通过构建氨单加氧酶基因(amoA)克隆文库,研究在红壤蔬菜田上只施用磷钾化肥(PK)、只施氮磷钾化肥(NPK)、施用腐熟有机肥(DNPK)和施用新鲜有机肥(FNPK)等4种不同施肥处理的土壤氨氧化细菌(AOB)和氨氧化古菌(AOA)群落多样性及与土壤脲酶活性的相关性。结果表明:施加有机肥处理(DNPK和FNPK)的蔬菜田土壤的AOB文库和AOA文库OTU数量和Shannon指数高于只施用无机肥(NPK和PK)处理的蔬菜田土壤;DNPK和FNPK处理的土壤优势AOB菌群为多形亚硝化叶菌(Nitrosolobus multiformis),比例分别为88.5%和68.5%,NPK和PK处理的土壤优势AOB菌群为亚硝化单胞菌属(Nitrosospirasp.),比例分别为54.8%和65.5%;DNPK、FNPK、NPK和PK处理土壤优势AOA菌群均为阿伯丁土壤亚硝化细杆菌侯选种(CandidatusNitrosotalea devanaterra),比例分别为90.9%、84.4%、77.8%和45.2%;施加有机肥处理(DNPK和FNPK)的土壤脲酶活性和氨氧化微生物的多样性指数都高于只施用无机肥处理(NPK和PK);AOA群落多样性指数与土壤脲酶活性呈显著正相关,而AOB群落多样性与土壤脲酶活性相关性不显著。总体来看,有机肥比无机肥处理提高了AOA和AOB群落多样性,且AOA在红壤蔬菜田土壤氨氧化过程中起着更为重要的作用。     

Effect of hay type on cecal and fecal microbiome and fermentation parameters in horses

The effect of hay type on the microbiome of the equine gastrointestinal tract is relatively unexplored. Our objective was to characterize the cecal and fecal microbiome of mature horses consuming alfalfa or Smooth Bromegrass (brome) hay. Six cecally cannulated horses were used in a split-plot design run as a crossover in two periods. The whole plot treatment was ad libitum access to brome or alfalfa hay fed over two 21-d acclimation periods with subplots of sampling location (cecum and rectum) and sampling hour. Each acclimation period was followed by a 24-h collection period where cecal and fecal samples were collected every 3 h for analysis of pH and volatile fatty acids (VFA). Fecal and cecal samples were pooled and sent to a commercial lab (MR DNA, Shallowater, TX) for the amplification of the V4 region of the 16S rRNA gene and sequenced using Illumina HiSeq. The main effects of hay on VFA, pH, and taxonomic abundances were analyzed using the MIXED procedure of SAS 9.4 with fixed effects of hay, hour, location, period, and all possible interactions and random effect of horse. Alpha and beta diversities were analyzed using the R Dame package. Horses fed alfalfa had greater fecal than cecal pH (P ≤ 0.05), whereas horses fed brome had greater cecal than fecal pH (P ≤ 0.05). Regardless of hay type, total VFA concentrations were greater (P ≤ 0.05) in the cecum than in feces, and alfalfa resulted in greater (P ≤ 0.05) VFA concentrations than brome in both sampling locations. Alpha diversity was greater (P ≤ 0.05) in fecal compared with cecal samples. Microbial community structure within each sampling location and hay type differed from one another (P ≤ 0.05). Bacteroidetes were greater (P ≤ 0.05) in the cecum compared with the rectum, regardless of hay type. Firmicutes and Firmicutes:Bacteroidetes were greater (P ≤ 0.05) in the feces compared with cecal samples of alfalfa-fed horses. In all, fermentation parameters and bacterial abundances were impacted by hay type and sampling location in the hindgut.     

Isolation and Identification of Pathogenic Bacteria Causing Dairy Cow Mastitis in Liaoning and Drug Sensibility Test of Pathogenic E.coli

In order to understand the main bacterial pathogen species causing dairy cow mastitis in Liaoning, as well as the characteristics of drug sensitivity of the pathogenic E.coli,the milk samples from 75 dairy cows with clinical manifestations for mastitis in certain large-scale dairy farm in Liaoning were collected.The bacteria in milk were cultured and isolated with biochemical methods and in vitro drug sensitivity tests were processed with the isolated E.coli strains.The results showed that the main bacterial pathogen for dairy cow mastitis were E.coli（separation rate 58.7%),S.aureus（64.0%）and S.agalactiae（54.7%),and multiple infection including double and triple infection were identified.The drug sensitivity tests on the isolated E.coli indicated that the E.coli isolates were highly resistant to sulfonamides（resistance rate>85%）and chloramphenicol（resistance rate>30%),and they were relatively low resistant to ampicillin（9.5%),ciprofloxacin（9.5%),ceftiofur（7.1%）and ofloxacin（4.8%).The results was able to provide reliable theoretical basis for prevention and control of dairy cow mastitis in Liaoning area.