Evaluation of a high-throughput nucleic acid extraction method for the detection of Mycobacterium avium subsp. paratuberculosis in bovine fecal samples by PCR

Johne’s disease (paratuberculosis) is an economically important disease of cattle worldwide. The disease is caused by Mycobacterium avium subsp. paratuberculosis (MAP), a fastidious gram-positive bacterium. PCR is increasingly used in diagnostic laboratories for the detection of MAP in fecal samples given the rapid test turnaround time and sensitivity and specificity comparable to fecal culture. However, efficient extraction of DNA for sensitive detection of MAP by PCR is affected by the complex lipid-rich cell wall of MAP and the presence of PCR inhibitors in feces. We evaluated a high-throughput nucleic acid extraction method (MagMAX core nucleic acid purification kit with mechanical lysis module) in conjunction with an hspX gene PCR for the detection of MAP from bovine fecal samples, which resulted in correct identification of all negative (13 of 13) and positive (35 of 35) proficiency test samples obtained from the National Veterinary Services Laboratories. In addition, all 6 negative and 50 of 51 positive diagnostic specimens tested were categorized correctly.     

影响猪采食行为的肠道微生物种类鉴别

动物主要通过采食来获取能量和所需营养物质,探索肠道菌群对猪采食行为的影响并研究其潜在的作用机制,可以为后续通过调控肠道菌群来改善猪采食提供理论基础。以210头商业杜洛克猪为研究对象,使用自动喂料器记录包括平均日采食时间(ADET)、平均日采食次数(ADEV)、平均日采食量(ADFI)等采食行为指标。140日龄时于肛门处收集粪便样品,通过16S rRNA基因V3-V4区测序获得试验猪肠道微生物结构与组成概况,采用Two-part模型以及共丰度组(CAGs)鉴别与猪采食行为相关的肠道微生物种类。结果显示：1)表型间相关性分析结果表明,ADET分别与ADEV(r=0.41,P<0.05)、RFI(r=0.32,P<0.05)呈显著正相关,ADFI分别与RFI(r=0.56,P<0.05)、平均日增重(ADG)(r=0.63,P<0.05)呈显著正相关,另外,RFI与背膘厚呈显著正相关(r=0.16,P<0.05)。2)CAGs分析中,主要包括瘤胃球菌科(Ruminococcaceae)、拟杆菌目(Bacteroidales)以及颤螺菌属(Oscillospira...     

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。     

Structural changes in lignin of tropical woods during digestion by termite, <Emphasis Type="Italic">Cryptotermes brevis</Emphasis>

Wood samples of apitong (Dipterocarpus grandiflorua) and ilang-ilang (Ilang-Ilang C. dadloyi) and feces of termites [Cryptotermes brevis (Walker)] fed on these woods were collected from University of the Philippines, Los Baňos. Lignin of each sample was isolated by Björkman’s procedure. There was no significant difference in ¹H nuclear magnetic resonance (NMR) spectra or in the methoxyl content between Björkman lignins from original woods and termite feces. Differences were detected in the contents of aliphatic and unconjugated phenolic hydroxyl groups, suggesting minor structural changes of lignin during digestion by termites. In addition, the ratio of syringyl to guaiacyl nuclei of Björkman lignin from termite feces determined by ¹H NMR spectra was higher than those from the original woods. The molar ratio of syringyl to guaiacyl nuclei of termite feces was higher than those from the original woods as determined by alkaline nitrobenzene oxidation. These results suggest that the structural changes of lignin in the termite gut are due to the insignificant formation of C-C linkages in guaiacyl nuclei. It was concluded that there were minor changes in the structural features of lignin under mostly anaerobic conditions, in contrast to the significant changes that occur through biological modification under aerobic conditions.     

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。     

蚕食药用价值开发现状与展望

综述了目前蚕幼虫、蚕蛹、蚕蛾和蚕沙等蚕资源在食用和药用价值开发中的最新研究进展,分析了存在的问题,并对蚕资源的进一步开发进行探讨。     

磷在棕壤中淋溶迁移特征研究

采用室内土柱淋溶模拟试验，研究相当于一年降雨条件下磷肥在棕壤土垂直方向上的淋溶迁移特征及鸡粪、秸秆对磷素迁移转化的影响。结果表明：土壤施肥量与速效磷、迟效态磷增加量呈极显著正相关。相关系数分别是0．99141。，0．99774；水溶性磷、Al—P与土壤速效磷相关性最好，相关系数分别达0．972和0．926，2500mg／kg鸡粪与25mg／kg磷肥配施能降低磷淋溶率，并能极显著提高土壤中速效磷的增加量，相当于单施50mg／kg的磷肥；秸秆与磷肥配合施用在短期内对土壤磷素的有效性影响不大，但可以减少磷素向下层迁移。     

一种用于畜禽粪便发酵的装置

以"一种用于畜禽粪便发酵的装置"为对象,阐述其工作原理、结构组成和功能,并对其发酵特点、发酵过程、发酵效果评价及等进行综述,为畜禽粪便的处理与利用提供参考.     

重庆市畜禽粪便年排放量的估算研究

畜禽养殖产生的粪便已成为我国农村面源污染的主要来源，因而畜禽粪便的排放量也越来越引起人们的重视。通过对重庆市畜禽养殖业进行调查，结合国内外有关研究，确定重庆市畜禽粪便年排放量估算方法和各种估算参数。以2001年为基准，估算出重庆市主要畜禽(猪、牛、羊、马、兔、家禽等)年粪尿排放量和粪尿中对环境产生污染的主要物质量(有机质、氮、磷、钾)，以便对重庆市畜禽粪便污染作一个定量的认识。估算结果为重庆市2001年畜禽粪尿排放7421万t，其中有机质为806万t，氮42.5万t、磷21.48万t、钾44.98万t。该研究结果可为重庆市工农业生产布局和环境污染治理提供决策依据。     

青海地区野生狐狸棘球绦虫感染情况调查研究

野生狐狸是棘球绦虫的主要终末宿主,在棘球蚴病的传播中有重要作用。为了解青海省狐狸中棘球绦虫感染状况,我们用棘球绦虫粪抗原检测试剂盒,对收集于青海省称多县、贵南县、祁连县的野生狐狸粪便进行了检测。结果表明,称多县收集的129份狐狸粪便阳性15份,阳性率11.6%;祁连县收集的84份狐狸粪便阳性3份,阳性率3.6%;贵南县收集的47份狐狸粪便阳性0份。共收集的260份狐狸粪便中阳性18份,平均阳性率7.5%。青海省野生狐狸中有棘球绦虫感染,在棘球蚴防治中应加强野生终末宿主的防治。