艾纳香内生真菌抗细菌和炭疽菌的活性研究

为获得丰富的艾纳香内生真菌资源,筛选出抗性生防菌株,用于活性次生代谢产物的发现,通过水琼脂法对海南产艾纳香进行内生真菌分离,通过ITS序列分析,对其进行鉴定和分析。采用滤纸片法评价内生真菌发酵产物对细菌的抑制活性,采用平板对峙培养法评价艾纳香内生真菌对植物炭疽菌的拮抗作用。本研究共分离鉴定出191株内生真菌,分属于27属,45种,其中优势菌属为Diaporthe、Paraphoma、Ectophoma、Penicillium、Talaromyces、Hypoxylon、Phomopsis、Colletotrichum和Fusarium。抗细菌活性结果显示,Clonostachys、Fusarium和Diapothe属活性较强。抗炭疽病菌试验显示,Talaromyces、Ectophoma和Penicillium属活性较强。本研究初步探讨了海南产艾纳香内生真菌种类的多样性,并首次对其进行了抗病原菌活性筛查,获得的活性菌株可作为微生物源菌剂开发的重要材料。     

Effect of multi-enzymes in combination with a direct-fed microbial on performance and welfare parameters in broilers under commercial production settings

The objective of this study was to determine the response of broilers to the combination of multi-enzymes and direct-fed microbial (DFM) under commercial production settings. A total of 7,000 1-day-old male broilers (Ross 308) were distributed over 10 pens (700 broilers/pen). Two dietary treatments were tested using complete randomized design, including a control diet and a test diet with addition of multi-enzymes (xylanase, amylase, and protease (XAP)] and DFM (a combination of spores from 3 strains ofBacillus amyloliquefaciens). Pelleted diets were offered ad libitum in 3 phases and water was freely available. During starter and grower phases (0 to 21 d), the enzyme and DFM combination resulted in improved FE (P < 0.05). During the finisher phase, higher feed intake and BW gain (P < 0.05) were observed for the test group. Overall, there were significantly higher feed intake, BW gain, and lower water-to-feed ratio in test group compared to the control group. This was related to improved (P < 0.05) modified production efficiency factor which was calculated based on final BW, survival rate, feeding period, and mortality-weight-corrected FCR. The test group had improved litter quality and a reduced foot-pad lesion score compared to the control. In addition, there was a tendency (P < 0.1) of reducingClostridium perfringens population in cecal digesta and higher lactic acid content in the ileal digesta, when expressed on an as-is basis, in the test group. In this study, we demonstrated that using a multi-enzymes and DFM combination in the diet for broilers can result in improved FE in starter/grower phases and animal welfare parameters, and lead to improved production efficiency under commercial settings.     

豆秸、花生秧和青贮玉米秸间的组合效应研究

本试验旨在研究豆秸、花生秧和青贮玉米秸间的组合效应。试验将豆秸、花生秧和青贮玉米秸分别以0∶100、20∶80、40∶60、60∶40、80∶20、100∶0比例进行两两组合,利用体外瘤胃发酵技术,分析不同组合对产气量及产气参数、pH、氨态氮、菌体蛋白的影响,计算各组合的单项组合效应值和综合组合效应值,进而筛选各饲料组合的适宜比例。结果表明：豆秸-花生秧、豆秸-青贮玉米秸、花生秧-青贮玉米秸组合在产气参数上差异显著(P<0.05),豆秸-花生秧和豆秸-青贮玉米秸均以20∶80时各产气参数最优,花生秧-青贮玉米秸以60∶40时各产气参数为最优;各组合对体外瘤胃液pH影响差异不显著(P>0.05);不同组合的氨态氮浓度差异显著(P<0.05),在39~64 mg/dL变化;豆秸-花生秧和花生秧-青贮玉米秸的菌体蛋白浓度随花生秧比例增多而增多,豆秸-青贮玉米秸的菌体蛋白浓度在两者比例为20∶80时最高。以多项组合效应评定指数评定各组合效应,豆秸与花生秧、青贮玉米秸均以20∶80的比例较为适宜,花生秧与青贮玉米秸比例以60∶40时较为适宜,综合组合效应指数均达到最大。     

祁连山不同植被类型土壤微生物群落多样性差异

土壤微生物是土壤生态系统的重要组成部分,在生态系统物质循环和能量转化中占有重要的地位。以祁连山中段不同海拔的4种植物群落,即垫状植被(cushion plants,CP)、高寒草甸(alpine meadow,AM)、沼泽化草甸(swamp meadow,SM)和高寒灌丛(alpine shrub,AS)作为实验样地,采用BIOLOG技术,探讨了土壤微生物群落多样性在不用海拔梯度形成的植被条件下的变化特征。结果表明,反映微生物活性的平均颜色变化率(average well color development,AWCD)的大小顺序为:沼泽化草甸高寒草甸高寒灌丛垫状植被,沼泽化草甸土壤微生物群落代谢活性最高;高寒草甸和沼泽化草甸土壤微生物群落碳源利用模式相似,土壤微生物群落的Shannon-Wiener物种丰富度指数(H)与土壤有机碳和全氮显著相关(P0.05),Simpson优势度指数(D)和Mc Intosh指数(U)与土壤全氮显著相关(P0.05);冗余分析表明,土壤有机碳、p H和全氮可能是土壤微生物利用碳源的控制因子。研究结果为进一步探讨高寒草甸与土壤微生物之间的关系奠定了基础。     

放牧对草地土壤微生物的影响

土壤质量是草地生态系统生产力可持续发展的关键所在。健康的土壤必定拥有丰富的物种多样性、活跃的生命体、高效的内部养分循环及较好的抗干扰能力。土壤微生物作为土壤系统中各种生物进程的主要推动者,其作用不可小觑。因此,我们可以利用土壤微生物的数量、群落结构和多样性等特征监测土壤质量。本文综述了放牧对草地土壤微生物数量、生物量、群落结构和多样性的影响。指出通过了解土壤微生物与放牧行为间的关系,如何评估土壤质量,选取适宜的放牧梯度,完善草地管理模式,并介绍了土壤微生物群落结构和多样性研究方法的利弊。解决两个关键问题：1）土壤微生物的各种特征是如何响应放牧行为的。2）如何通过这些指标评价土壤质量,然后提出合理的放牧模式,改善和维护草地的生产力。     

不同代谢葡萄糖水平对绵羊人工瘤胃发酵特性、微生物蛋白质浓度和产气参数的影响

本试验旨在通过人工瘤胃体外培养,研究不同代谢葡萄糖水平下的绵羊瘤胃发酵特性、微生物蛋白质浓度和产气参数。采用单因子试验设计,共设计4个代谢葡萄糖水平[125(A)、138(B)、153(C)、168 g/kg(D)]。体外试验所用瘤胃液采自4只安装有永久性瘤胃瘘管的绵羊。分别于培养0、2、4、6、8、12、24 h采集2 mL培养液用于分析。结果表明:1)8~24 h培养液pH随着代谢葡萄糖水平的提高而出现显著或极显著下降(P0.05或P0.01);氨氮浓度在2 h时D组显著高于A组(P0.05),而在6 h时A组显著高于其他3组(P0.05)。2)D组6h培养液细菌蛋白浓度显著高于A组(P0.05);随着代谢葡萄糖水平的提高,培养液丙酸、丁酸、总挥发性脂肪酸的浓度总体呈升高趋势,乙酸/丙酸呈下降趋势。3)随着代谢葡萄糖水平的提高,理论最大产气量极显著降低(P0.01),达1/2理论最大产气量的时间极显著缩短(P0.01);潜在产气量无显著变化(P0.05),24 h产气量和产气速率常数分别显著或极显著下降和上升(P0.05或P0.01)。结果提示,提高代谢葡萄糖水平,可以提高丙酸、总挥发性脂肪酸的浓度,同时可为绵羊提供较多的生糖前体物质。     

Distribution and Diversity of Salmonella Strains in Shipments of Hatchling Poultry,United States, 2013

Multistate outbreaks of salmonellosis associated with live poultry contact have been occurring with increasing frequency. In 2013, multistate outbreaks of salmonellosis were traced back to exposure to live poultry, some of which were purchased at a national chain of farm stores (Farm store chain Y). This study was conducted at 36 stores of Farm store chain Y and was concurrent with the timing of exposure for the human outbreaks of salmonellosis in 2013. We used environmental swabs of arriving shipment boxes of hatchling poultry and shipment tracking information to examine the distribution, diversity and anti‐microbial resistance of non‐typhoidal Salmonella (NTS) across farm stores and hatcheries. Isolates recovered from shipment boxes underwent serotyping, anti‐microbial resistance (AMR) testing and pulsed‐field gel electrophoresis (PFGE). Postal service tracking codes from the shipment boxes were used to determine the hatchery of origin. The PFGE patterns were compared with the PFGE patterns of NTS causing outbreaks of salmonellosis in 2013. A total of 219 hatchling boxes from 36 stores in 13 states were swabbed between 15 March 2013 and 18 April 2013. NTS were recovered from 59 (27%) of 219 hatchling boxes. Recovery was not significantly associated with species of hatchlings, number of birds in the shipment box, or the presence of dead, injured or sick birds. Four of the 23 PFGE patterns and 23 of 50 isolates were indistinguishable from strains causing human outbreaks in 2013. For serotypes associated with human illnesses, PFGE patterns most frequently recovered from shipment boxes were also more frequent causes of human illness. Boxes positive for the same PFGE pattern most frequently originated from the same mail‐order hatchery. Only one of 59 isolates was resistant to anti‐microbials used to treat Salmonella infections in people. This study provides critical information to address recurrent human outbreaks of salmonellosis associated with mail‐order hatchling poultry.     

6 个不同品种荔枝果肉内生细菌群落多样性的高通量测序分析

【目的】内生菌在植物生长发育中发挥重要作用,分析不同品种荔枝果肉内生细菌群落结构及多样性,挖掘和利用潜在荔枝内生菌种资源,为进一步研究荔枝与内生菌互作关系提供参考依据。【方法】以 6 个不同品种荔枝的果肉组织为材料,PCR 扩增 16S rDNA 的 V3~V4 区,利用 MiSeq 高通量测序技术分析果肉组织内生细菌在门、科、属和种分类水平上的群落组成及优势菌群,结合 Alpha 多样性分析评估内生细菌群落多样性。【结果】对 6 个品种荔枝果肉内生细菌 16S rDNA 的 V3~V4 区进行扩增,共获得 2 139 086 条序列,其平均长度为 376 bp。不同品种荔枝果肉内生细菌群落在相对丰度和多样性上无显著差异。在门分类水平,变形菌门为主要优势细菌门类,其次为放线菌门。在科、属和种分类水平上,不同品种荔枝内生菌群落结构相似,但相对丰度占比不同。差异菌群分析结果显示,假单胞菌属（Pseudomonas）、果胶杆菌属（Pectobacterium）、苍白杆菌属（Ochrobactrum）和迪基氏菌属（Dickeya) 在优良品种‘鹅蛋荔’‘糯米糍’和‘观音绿’中显著富集,根瘤菌属（Bradyrhizobium）和变形菌属（Snodgrassella）只在‘鹅蛋荔’中富集。根据菌群相对丰度进行聚类分析,‘观音绿’‘糯米糍’和‘鹅蛋荔’的相似性更高。Alpha 多样性分析结果表明,6 种荔枝果肉内生菌群仅在丰度和多样性上存在差异,但未达到显著水平。【结论】该研究首次分析了 6 个不同品种荔枝果肉内生细菌的群落结构,并在优良荔枝品种中发现特有的微 生物类群富集现象,可为后续提升荔枝品质、开发利用内生菌提供理论依据和重要参考。     

不同粒度猫尾草对羔羊体外发酵特性和微生物数量的影响

通过Rusitec-s体外发酵系统研究不同粒度猫尾草对羔羊体外发酵参数与微生物数量的影响,选取6种不同粒度(1.00、2.36、3.35、4.75、8.00和12.50 mm)的猫尾草作为试验饲粮的纤维来源,按照猫尾草的粒度将试验分为6个处理组,每个处理4个重复,利用Rusitec-s型人工瘤胃模拟装置进行48 h体外发酵,测定养分降解率、发酵参数及发酵液中6种微生物菌群的拷贝数。结果表明：与其他处理组相比,2.36 mm组显著提高了体外中性洗涤纤维降解率(IVNDFD)、丁酸、异丁酸、戊酸、异戊酸摩尔比和溶纤维丁酸弧菌(B. f)数量(P<0.05),且显著降低总产甲烷菌(TMe)数量(P<0.05)。总菌(TB)与氨态氮(NH3-N)浓度、乙酸摩尔比呈正相关(R²=0.84,P=0.038;R²=0.88,P=0.021);B. f数量与丙酸摩尔比呈正相关(R²=0.82,P=0.045),且与pH呈负相关(R²=-0.98,P=0.001);TMe数量与体外粗蛋白质降解率(IVCP...     

哺乳期幼驼与成年母驼粪便菌群多样性分析

【目的】探究准噶尔双峰驼哺乳期幼驼和成年母驼粪便菌群结构组成及多样性变化,为幼驼肠道健康发育提供重要依据。【方法】选取6~7岁体况相近的健康成年母驼12峰,以及出生日期相近（3月龄）的哺乳期母幼驼12峰,母驼与幼驼在相同饲养环境下饲养。采用直肠取粪法采集粪样,用于粪便内容物细菌16S rDNA的V3-V4区测序,并对测序数据进行相应分析。【结果】母驼粪便菌群Chao1和Shannon指数均显著高于幼驼（P<0.05）;在门水平上,母驼和幼驼粪便中位于前十的菌均为厚壁菌门、拟杆菌门、疣微菌门、变形菌门、软壁菌门、螺旋体门、TM7、放线菌门、蓝藻菌门和纤维杆菌门,其中幼驼粪便中变形菌门和软壁菌门丰度均极显著高于母驼（P<0.01）;在科水平上,母驼与幼驼粪便中位于前十的菌均为瘤胃球菌科、消化链球菌科、毛螺菌科、梭菌科、克里斯滕森菌科、理研菌科、疣微菌科、拟杆菌科、艰难杆菌科和普雷沃氏菌科,其中母驼粪便中消化链球菌科和梭菌科的丰度均极显著高于幼驼（P<0.01）,而幼驼粪便中毛螺菌科的丰度极显著高于母驼（P<0.01）;在属水平上,母驼与幼驼粪便中位于前十的菌属均为梭菌属、颤螺旋菌属、瘤胃球菌属、梭菌科-梭菌属、阿克曼菌属、5-7 N15、CF231、苏黎世杆菌属、梭菌科-SMB53和密螺旋体菌属,其中母驼粪便中的梭菌属、梭菌科-梭菌属和苏黎世杆菌属的丰度均极显著高于幼驼（P<0.01）。【结论】成年母驼粪便菌群多样性显著高于哺乳期幼驼,且稳定性较好;母驼和幼驼的粪便在门、科、属水平下的主导菌属种类均一致,但是丰度均存在差异。