Effect of transferring lignocellulose-degrading bacteria from termite to rumen fluid of sheep on in vitro gas production,fermentation parameters,microbial populations and enzyme activity

The digestive tract of termite(Microcerotermes diversus) contains a variety of lignocellulose-degrading bacteria with exocellulases enzyme activity, not found in the rumen, which could potentially improve fiber degradation in the rumen. The objectives of the current study were to determine the effect of inoculation of rumen fluid(RF) with three species of bacteria isolated from termite digestive tract, Bacillus licheniformis, Ochrobactrum intermedium, and Microbacterium paludicola, on in vitro gas production(IVGP), fermentation parameters, nutrient disappearance, microbial populations, and hydrolytic enzyme activities with fibrous wheat straw(WS) and date leaf(DL) as incubation substrate. Inoculation of RF with either of three termite bacteria increased(P0.05) ammonia-N concentration compared with the control group(free of termite gut bacteria). Termite bacteria inoculation had no effect(P0.05) on gas production characteristics, dry matter, organic matter and neutral detergent fiber disappearance, pH, and concentration and composition of volatile fatty acids. Population of proteolytic bacteria and protozoa, but not cellulolytic bacteria, were increased(P0.05) when RF was inoculated with termite bacteria with both WS and DL substrates. Inoculation of RF with termite bacteria increased protease activity, while activities of carboxymethyl-cellulase, microcrystalline-cellulase, α-amylase and filter paper degrading activity remained unchanged(P0.05). Overall, the results of this study indicated that transferring lignocellulose-degrading bacteria, isolated from digestive tract of termite, to rumen liquid increased protozoa and proteolytic bacteria population and consequently increased protease activity and ammonia-N concentration in vitro, however, no effect on fermentation and fiber degradation parameters were detected. These results suggest that the termite bacteria might be rapidly lysed by the rumen microbes before beneficial effects on the rumen fermentation process could occur.     

植物内生肠杆菌对狗牙根耐盐性的调控研究

以从海滨雀稗中分离得到的内生肠杆菌为材料，采用种子萌发与盆栽试验两种方式研究植物内生细菌对狗牙根耐盐性的影响。通过对狗牙根种子和幼苗接种单一内生细菌和混合内生细菌，并测定种子发芽率、苗长、根长、生物量等生长指标与叶绿素含量、相对含水量、相对电导率和叶片中钠离子、钾离子的含量综合评价植物内生细菌对狗牙根耐盐能力的调控。结果表明，在150 mmol·L^-1盐胁迫下处理14 d后，接种混合菌液B3014比接种单一菌液B30更能提升狗牙根种子的萌发率、对盐胁迫下胚根及胚芽长度的增长也较显著（P<0.05）。在成坪期，与空白对照相比，接种内生细菌可以提升200 mmol·L^-1盐胁迫下狗牙根的成坪质量，增加地上、地下生物量的积累、苗长与根长生长量，并提升植株叶绿素含量与叶片相对含水量，降低细胞膜破损程度。在接种内生细菌处理中，接种混合菌液B3014可以使狗牙根在盐胁迫下受到的损伤显著低于（P<0.05）接种单一菌液B30。与此同时，接种混合菌液B3014对狗牙根叶片中钠离子含量降低程度高于接种单一菌液B30，同样，接种混合菌液B3014也能更好地积累盐胁迫下狗牙根叶片内钾离子含量，增加盐胁迫下狗牙根的耐盐能力。因此，接种混合菌液B3014对盐胁迫下狗牙根种子的萌发和成坪后的耐盐能力的提升高于接种单一菌液B30。探讨植物内生细菌对狗牙根耐盐能力的调控，为提高草坪草在盐碱环境中的应用提供了新思路。     

马铃薯与玉米复合种植对土壤化感物质及土壤细菌群落结构的影响

为探究马铃薯与玉米复合种植对化感物质积累与细菌群落结构的影响,分析轮作、间作缓解连作障碍的机制,本研究以马铃薯连作、玉米连作、马铃薯||玉米间作、马铃薯-玉米轮作第8年的土壤为对象,利用GC-MS测定土壤中化感物质含量,并采用Illumina Miseq高通量测序技术对土壤细菌16Sr DNA V4-V5区域进行测序,分析土壤中细菌多样性和群落结构的变化,并对化感物质和优势菌属进行相关性分析。结果表明:玉米连作和马铃薯连作会导致化感物质的积累,玉米连作土壤积累了更多的油酸、亚油酸、花生酸、木焦油酸等脂肪酸,马铃薯连作土壤积累了更多的硬脂醇、二十烷醇等脂肪醇类物质。轮作降低了大部分化感物质的积累,间作降低的化感物质种类相对轮作较少。不同种植方式下土壤细菌群落结构发生了显著变化,相对于连作,间作和轮作Ace指数和Chao指数显著升高。在门水平上,轮作土壤放线菌丰度显著高于马铃薯连作土壤,间作土壤拟杆菌门丰度显著低于玉米连作土壤,两种连作土壤中酸杆菌门丰度都较轮作显著升高。在属水平上,一些有益细菌如节杆菌属、溶杆菌属等在复合种植土壤中相对丰度更高。通过相关性分析发现土微菌属、小梨形菌属与脂肪醇类物质呈显著正相关,黄杆菌属、溶杆菌属、微杆菌属等与脂肪酸类物质呈显著负相关。马铃薯与玉米复合种植降低了化感物质在土壤中的积累,从而抑制了土壤细菌丰度的降低,提高了有益菌属丰度,消减了连作障碍。     

低聚半乳糖干预缓解肠道炎症的研究进展

肠道炎症已成为我国社会健康的难题和挑战，其发病率在我国迅速增长。肠道炎症发病原因复杂，目前尚缺乏有效的缓解药物，因此加强肠道炎症有效缓解物质的研发至关重要。低聚半乳糖（galactooligosaccharide，GOS）是一种食疗益生性较优的乳源功能性低聚糖，能够有效促进肠道内益生菌的增殖，改变肠道菌群结构，刺激免疫应答，进而改善肠黏膜屏障功能，缓解肠道炎症。本文综述近年来国内外有关肠道炎症及GOS干预缓解肠道炎症作用的研究进展，并对其应用前景进行展望，为此领域的研究提供科学依据。     

规模化蛋鸡场病原菌溯源与生物安全防控研究

细菌病严重威胁蛋鸡健康,以抗生素为主导的细菌病传统防控方式会导致病原菌耐药性严重、鸡蛋药物残留,影响蛋品安全。要保证蛋鸡产蛋期不用抗菌药物而又能生产出"无菌、无抗"的鸡蛋,是国内外技术难题。本团队以规模化蛋鸡场全封闭鸡舍为研究对象,基于PFGE、MLST、全基因组测序等技术,探明了蛋鸡病原菌的种类及基因组特征;对病原菌的耐药性研究表明,蛋鸡病原菌多重耐药严重,依靠药物防控的方式亟待转变;对规模化蛋鸡场细菌溯源研究结果表明,鸡舍生物媒介(鼠、苍蝇等)和非生物媒介(空气、饮水、饲料、鸡粪等)是蛋鸡病原菌的重要传染来源和传播途径;对规模化蛋鸡场生物安全量化评价结果表明,外部和内部生物安全权重高达92.2%。根据溯源和生物安全权重分析结果,提出了在规模化全封闭蛋鸡舍将蛋鸡细菌病从药物防控转变为生物安全防控的新策略;创新了规模化蛋鸡场细菌病防控的系统技术,实现蛋鸡规模养殖整个产蛋期不用抗生素,为生产"无菌、无抗"安全鸡蛋提供技术支撑。     

亮斑扁角水虻形态观察及诱产卵菌的筛选

【目的】 研究引进亮斑扁角水虻在室内饲喂条件下的生活史、成虫种群雌雄比例变化规律,对其卵表及幼虫肠道中诱成虫产卵菌进行筛选和鉴定,为亮斑扁角水虻在本地区的规模养殖和综合利用提供理论依据。【方法】 引进亮斑扁角水虻,在室内光照条件下饲喂4代,用直接观察法记录亮斑扁角水虻在不同饲喂条件下的生活史和生活习性,显微拍照记录水虻各龄期的形态特征;用稀释涂布平板及平板划线的方法,对亮斑扁角水虻卵携带菌及其幼虫肠道菌进行分离纯化,筛选可诱导产卵的菌株,并鉴定。【结果】 与内地相比,亮斑扁角水虻在新疆饲养周期稍有延长,但其发育完好,能够在本地区进行世代饲养;观察并记录了其卵、幼虫、蛹、成虫及其其生殖器形态;筛选到对亮斑扁角水虻雌性成虫有诱导产卵效果的两株菌e1、Y3,其发酵液处理组平均产卵量分别为对照组的6.26和7.40倍,两株菌皆为革兰氏阳性菌,16S RNA测序初步鉴定e1为暹罗芽孢秆菌,Y3为节杆菌属,且Y3为1种潜在的新菌。【结论】 引进亮斑扁角水虻在新疆能够周年循环养殖,具有规模生产及综合开发利用潜力。     

棉花苗期根腐类病害生防细菌的筛选与鉴定

【目的】从新疆棉田中筛选对棉花苗期根腐类病害具有较强拮抗作用的生防细菌,为棉花苗期根腐类病害生物防治奠定基础。【方法】采用平板对峙法,测定实验室保存的多株生防细菌的抑菌能力,通过田间试验,验证其防治棉花苗期根腐类病害的效果。对生防细菌的16S rDNA基因序列与gyrA基因序列分别测序,通过BLAST在线比对,利用MEGA 7.0构建系统发育树,确定该生防菌株的分类地位。【结果】生防细菌TWt34、HWt34对棉花苗期根腐类病害(立枯病和红腐病)防治效果分别为76.74%和78.43%;在平板对峙对立枯丝核菌(Rhizoctonia solani) 和拟轮枝镰孢霉(Fusarium verticilliodes)的抑菌宽度分别为8.29和6.60 mm,具有较强拮抗作用,对尖孢镰刀菌(Fusarium oxysporum)和大丽轮枝菌(Verticillium dahliae)的抑菌宽度分别为7.70 和13.15 mm,也具有明显抑制作用。16S rDNA与gyrA基因序列鉴定表明生防菌菌株TWt34、HWt34同属于莫海威芽孢杆菌(Bacillus mojavensis)。【结论】筛选出的2株莫海威芽孢杆菌TWt34、HWt34对棉花4种主要病害的病原菌具有较强的抑制作用,能够有效防治2种棉花苗期根腐类病害(立枯病和红腐病)。     

Enhancing rotifer Brachionus calyciflorus population growth with commercial probiotics

The effect of a commercial probiotic (NanoCrusta, Altacrusta, Mexico City, Mexico) on the growth performance of Brachionus calyciflorus Pallas, 1766, was evaluated. In a first approach, probiotics were supplied in four densities (2.0 × 10³, 1.1 × 10⁵, 2.1 × 10⁵ and 2.1 × 10⁶ cells/ml), alone and in combination with Chlorella vulgaris (1 × 10⁶ cells/ml). The test rotifer did not grow on the probiotic alone. However, when probiotics + C. vulgaris were added, the maximum densities (D_max; ind/ml) and population growth rates (r) observed were higher. In the second experiment, probiotics were supplied at five higher densities (2.1 × 10⁶, 4.2 × 10⁶, 8.5 × 10⁶, 1.7 × 10⁷ and 3.4 × 10⁷ cells/ml) with C. vulgaris and a control treatment with only C. vulgaris (probiotic‐free). Treatments supplied with probiotics between 2.1 × 10⁶ and 1.7 × 10⁷ cells/ml showed significantly higher D_max and r than the control treatment. The results showed a positive effect of probiotic bacteria when supplied with C. vulgaris. The best outcome showed a D_max 2.16 times and an r 1.63 times higher than the density of the control treatment. Growth rates were higher in the treatments with probiotics compared to the control. We conclude that application of NanoCrusta is feasible to improve B. calyciflorus production, but the effects need to be tested in larger scales.     

Effects of beneficial bacteria on biomass,photosynthetic parameters and cell composition of the microalga Muriellopsis sp. adapted to grow in seawater

The present study evaluated the effects of five bacterial isolates from the microalga Muriellopsis sp. on the microalgal biomass concentration (g/L), specific growth rate (K), photosynthetic quantum yield (QY), non‐photochemical quenching (NPQ), maximum relative electron transport rates (rETR max) and levels of nutritional components such as carbohydrates, lipids, proteins and carotenoids in Muriellopsis sp. cultivated in seawater under indoor and outdoor conditions. The bacterial strains were Microbacterium sp. Dom 1; Pseudomonas sp. Dom2; Microbacterium sp. Dom 3; Rhizobium sp. Dom 4; and Dietzia sp. Dom 5, as well as the Pseudoalteromonas sp. (SLP1), a bacterium known to promote the growth of microalgae. At the laboratory level, inoculation of the strains SLP1, Dom 1, Dom 3 and Dom 5 resulted in higher specific growth rates and biomass of the microalga. Subsequently, the best results were obtained with a combination of the selected bacterial strains (Dom1, Dom 3, Dom5 and SLP1) under outdoor culture conditions improved the biomass, proteins, carbohydrates and total carotenoids in 22.12%, 48.28%, 19,25% and 48.27%, respectively, compared to cultures without the incorporation of the selected bacteria (control), while no effects were observed on the photosynthetic parameters. Thus, it was demonstrated that positive associations of the selected bacterial strains played an important role in the production of Muriellopsis sp. acclimatized to grow in seawater. This is of crucial importance, especially in desert areas, where solar radiation is high and freshwater is a limited resource.     

规模化畜禽养殖场粪便中多重耐药菌分离鉴定及其耐药特征

为了解禽畜粪便中多重耐药菌的污染特征,本研究对鸡粪、牛粪、猪粪和有机肥这4种不同样品中多重耐药菌进行了计数分析,并对不同来源的多重耐药菌株进行了分离和纯化,进一步开展了基于16S rDNA序列比对的分子生物学鉴定以确定其种属地位,以及基于药敏试验的耐药性特征分析。结果表明:不同养殖动物粪便中四环素、恩诺沙星、磺胺甲恶唑和泰乐菌素4种抗生素多重耐药菌的绝对数量和相对数量排序均为鸡粪>猪粪>牛粪,粪源有机肥中可培养的多重耐药菌的绝对数量和相对数量在堆肥后均有所下降。通过对耐药菌株鉴定和分类学分析,发现禽畜粪便中多耐药菌的菌门集中分布在Proteobacteria、Firmicutes和Actinobacteria,多重耐药菌的优势菌属为Escherichia、Corynebacterium、Kurthia;而有机肥样品中多重耐药菌的优势菌属为Staphylococcus、Glutamicibacter。菌株的药敏试验表明,3类动物粪污中的多重耐药菌都对红霉素、四环素有着较高的耐药率,均高于80%,而对阿米卡星这种抗生素表现出较好的敏感性,其耐药率低于30%。通过对Ⅰ、Ⅱ类整合子基因盒的扩增,发现PCR产物的大小从0.8 kb到1.8 kb不等,基因盒ad A2、dfr A17、dfr A1及sat2在养殖场粪污中检出率均较高。