石油降解菌筛选及应用研究

从石油污染土壤中分离到一株新的石油降解菌,对该石油降解菌进行了鉴定,经16SrDNA测序鉴定并结合生理生化实验结果确定其为阴沟肠杆菌。该菌可以将石油作为唯一碳源进行生长代谢,实验结果表明:在菌株投加量为1%时降解效果最佳,降解率可达到26.41%。     

大米草中耐盐乳酸菌的分离鉴定及对大米草青贮品质的影响

本试验旨在研究大米草（Spartina anglica Hubb.）中耐盐乳酸菌的分离鉴定及对大米草青贮品质的影响，以期为大米草青贮利用提供技术支持和理论依据。试验采用传统培养法筛选出耐盐浓度为10%的乳酸菌菌株，通过生理生化及16S rDNA序列分析法综合鉴定筛选的菌株，并将其添加到大米草原料中进行青贮效果观察。结果表明：分离获得的42株乳酸菌菌株，耐盐溶度最高为10%的4株乳酸菌均为同型发酵的植物乳杆菌（Lactobacillus plantarum），命名为C1、C2、C3和C4。4株乳酸菌均能提高大米草的青贮发酵品质，其中C4显著提高青贮料的可溶性碳水化合物和粗脂肪含量（P<0.05），显著降低青贮料的氨态氮含量（P<0.05）；另外，C4处理组的干物质含量和乳酸含量显著高于其他3个菌株处理组（P<0.05），中性洗涤纤维、酸性洗涤纤维、半纤维素和丁酸含量显著（P<0.05）低于其他3个菌株处理组，青贮效果最佳。     

Long-term fertilization effects on active ammonia oxidizers in an acidic upland soil in China

The effects of long-term fertilization of acidic soils on ammonia-oxidizing archaea (AOA) and bacteria (AOB) communities and its ecological implications remain poorly understood. We chose an acidic upland soil site under long-term (27-year) fertilization to investigate ammonia oxidizer communities under four different regimes: mineral N fertilizer (N), mineral NPK fertilizer (NPK), organic manure (OM) and an unfertilized control (CK). Soil net nitrification rates were significantly higher in OM soils than in CK, N or NPK soils. Quantitative analysis of the distribution of amoA genes by DNA-based stable isotope probing revealed that AOA dominate in CK, N and NPK soils, while AOB dominate in OM soils. Denaturing gradient gel electrophoresis and clone library analyses of amoA genes revealed that Group 1.1a-associated AOA (also referred to as Nitrosotalea) were the most dominant active AOA population (>92%), while Nitrosospira Cluster 3 and Cluster 9 were predominant among active AOB communities. The functional diversity of active ammonia oxidizers in acidic soils is affected by long-term fertilization practices, and the responses of active ammonia oxidizers to mineral fertilizer and organic manure are clearly different. Our results provide strong evidence that AOA are more highly adapted to growth at low pH and low substrate availability than AOB, and they suggest that the niche differentiation and metabolic diversity of ammonia oxidizers in acidic soils are more complex than previously thought.     

Investigation of Interspecies Interactions within Marine Micromonosporaceae Using an Improved Co-Culture Approach

With respect to bacterial natural products, a significant outcome of the genomic era was that the biosynthetic potential in many microorganisms surpassed the number of compounds isolated under standard laboratory growth conditions, particularly among certain members in the phylum Actinobacteria. Our group, as well as others, investigated interspecies interactions, via co-culture, as a technique to coax bacteria to produce novel natural products. While co-culture provides new opportunities, challenges exist and questions surrounding these methods remain unanswered. In marine bacteria, for example, how prevalent are interspecies interactions and how commonly do interactions result in novel natural products? In an attempt to begin to answer basic questions surrounding co-culture of marine microorganisms, we have tested both antibiotic activity-based and LC/MS-based methods to evaluate Micromonosporaceae secondary metabolite production in co-culture. Overall, our investigation of 65 Micromonosporaceae led to the identification of 12 Micromonosporaceae across three genera that produced unique metabolites in co-culture. Our results suggest that interspecies interactions were prevalent between marine Micromonosporaceae and marine mycolic acid-containing bacteria. Furthermore, our approach highlights a sensitive and rapid method for investigating interspecies interactions in search of novel antibiotics, secondary metabolites, and genes.     

不同补水方式下砂壤土渗滤系统对硝态氮去除效果

在水资源短缺的北京地区利用再生水回补城市河湖,一方面对于水资源的可持续利用有着十分重要的作用,另一方面也可能带来地下水环境的潜在污染风险.该文采用100 cm砂壤土柱模拟(河湖岸底)土地渗滤系统,设置定水头淹水、交替淹水落干、定流速补水和侧向补水4种不同再生水回补方式,研究再生水中硝态氮(NO3-N)在土地渗滤系统中的去除效果和迁移转化规律.结果表明,当水力负荷在0.25~2.65 cm/d范围内时,渗滤系统对NO3-N的去除率随着水力负荷的增大而减小;侧向补水方式下渗滤系统对NO3-N的去除效果最优,平均去除率高达96.1%.在定水头淹水和侧向补水方式下,系统对NO3-N的去除主要发生在土柱的上部,而交替淹水落干和定流速补水条件下,土柱中下部对NO3-N也有一定的去除作用.渗滤系统对NO3-N的去除主要取决于系统内部微生物的分布情况,土层中的反硝化细菌数量越大,该土层对NO3-N的去除率就越高.当水温在15~32℃范围内变化时,定水头淹水和交替淹水落干补水方式下,系统对NO3-N的去除率与温度分别呈指数和幂函数关系.该研究表明土地渗滤系统可实现再生水的进一步净化处理,可为再生水安全回补河湖提供参考.     

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。     

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。     

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。     

Effects of Lactic Acid Bacteria and Fructooligosaccharides on Growth Performance,Nutrient Digestibility and Serum Biochemical Indices of Arctic Foxes (Alopex lagopus) during Early Growth Period北大核心CSCD

This experiment was conducted to study the effects of dietary lactic acid bacteria (LAB) and fructooligosaccharides (FOS) on growth performance, nutrient digestibility and serum biochemical indices of Arctic foxes (Alopex lagopus) during early growth period. Sixty male post-litter Arctic foxes with the average body weight of (1.16±0.15) kg and at the age of (45±3) days were randomly assigned to 4 groups with 3 replicates per group and 5 foxes per replicate. The foxes were fed the following diets, respectively: basal diet (control group), basal diet+3×109 CFU / kg LAB (LAB group), basal diet+5 g / kg FOS (FOS group) and basal diet+3×109 CFU / kg LAB+5 g / kg FOS (LAB+FOS group). The experiment lasted for 30 days. The results showed as follows: 1) on the days 1 to 15, the diarrhea rate of foxes in the LAB group and LAB+FOS group was significantly lower than that in the control group and FOS group (P<0.05). On the days 16 to 30, the average daily gain of foxes in the LAB+FOS group was significantly higher than that in the control group and FOS group (P<0.05), but had no significant difference with LAB group (P>0.05); the diarrhea rate of foxes in the LAB+FOS group was significantly lower than that in the control group and FOS group (P<0.05), but had no significant difference with LAB group (P>0.05); compared with the FOS group, the feed to gain ratio of foxes in the LAB+FOS group was significantly decreased (P<0.05). 2) The dry matter digestibility of foxes in the LAB + FOS group was significantly higher than that in the control group and FOS group (P < 0.05), but had no significant difference with LAB group (P>0.05). There were no significant differences in nitrogen intake, fecal nitrogen, urine nitrogen, nitrogen deposition, net protein utilization and protein biological value among all groups (P>0.05). 3) Serum total protein (TP) content of foxes in the LAB+FOS group was significantly higher than that in the FOS group (P<0.05), but had no significant difference with control group and LAB group (P>0.05). Serum albumin (ALB) content of foxes in the LAB group was significantly higher than that in the control group and FOS group (P<0.05), but had no significant difference with LAB+ FOS group (P>0.05). Serum urea nitrogen (UN) content of foxes in the LAB+FOS group was significantly decreased compared with the control group (P < 0. 05). In conclusion, the combined supplementation of 3 × 109 CFU / kg LAB and 5 g / kg FOS in the diet is beneficial to reduce diarrhea rate, improve dry matter digestibility, improve body immune level, and promote the growth of Arctic foxes during early growth period. © 2023 Chinese Journal of Urology. All rights reserved.     

新疆核桃腐烂病拮抗菌的筛选及鉴定

为给核桃促生菌群的构建提供菌株材料,通过对新疆核桃拮抗菌的分离纯化,筛选出对核桃腐烂病病原菌具有拮抗作用且能在其根际稳定定殖的优质高效菌株。从阿克苏、和田、喀什等新疆核桃主产区的根际土壤中分离出拮抗菌30株,采用对峙试验法对其拮抗能力进行测定,选出拮抗菌14株。经耐利福平诱导后剩余7株。将其进行大田定殖试验后,最终筛选出7株拮抗菌。经16S r DNA鉴定,这7株拮抗菌属于3个属,分别为假单胞菌属Pseudomonas、芽孢杆菌属Bacillus、链霉菌属Streptomyces,其中假单胞菌属为优势种属,且拮抗效果及稳定性最好。