带属绦虫线粒体基因组全序列生物信息学分析

通过利用生物生物信息学方法分析带属绦虫线粒体DNA(mtDNA)碱基组成、基因结构、密码子利用、基因变异及其在分子系统进化研究中的应用价值等,以期为带属虫种线粒体功能基因组学研究、分子分类学研究及其疾病诊断提供重要依据。从NCBI GenBank中下载已测序的6种带属绦虫线粒体基因组(mt genome)全序列,以细粒棘球绦虫mtDNA序列作为参考序列(外群),用ClustalX软件(1.83)(http://www2.ebi.ac.uk/clustal w/)进行多重序列比对,用DNAStar软件进行序列差异性分析,用MEGA4软件的邻接法(Neighbor-joining method)构建进化树,核苷酸进化距离算法用最大似然法(Maxi mumlikelihood method),氨基酸进化距离算法用泊松校正法(Poissoncorrection method),进化树检验用自展法(Bootstrap test)。tRNA和2个非编码区RNA二级结构预测分别使用软件ARWEN和RNAstructure Version 4.6。结果显示,带属绦虫mtDNA为双链闭环分子,为13.3~13.7 kb;4种碱基成分中,T含量最高(超过47%),C含量最低(低于9%);共有36个按照相同顺序排列的编码基因,其中蛋白质编码基因有12个,tRNA编码基因有22个(其中编码丝氨酸-tRNA和亮氨酸-tRNA的基因有2个,其余18种tRNA分子各有1个),rRNA编码基因有2个(包括1个大亚基和1个小亚基)。基因组结构紧凑,基因间存在重叠区域,如nad4L和nad4基因;除广泛使用ATG作为起始密码子编码蛋氨酸,部分基因如多头绦虫nad6基因用GTG作为蛋白质翻译的起始密码子。线粒体tRNA长度为58~76 nt,二级结构多数呈典型的三叶草结构,少数呈D-loop结构;2个线粒体rRNA亚基基因rrnL和rrnS被trnC基因分隔开;线粒体基因组有2个大的非编码区,1个长非编码区(LNR)和1个短非编码区(SNR);线粒体基因组中蛋白编码基因之间的差异为5.7%~28.9%,其中cox1和nad4L基因比较保守,而nad5和nad6则变异较大,进化较快。结果表明,根据线粒体基因组序列绘制的带属绦虫分子进化树表明,亚洲带绦虫(T.asiatica)和牛带绦虫(T.saginata)间的亲缘关系最近,成为姊妹种,而多头绦虫(T.multiceps)与前两者的亲缘关系比猪带绦虫(T.solium)与前两者的关系更近,但与虫种的生物学特征存在一定差异。     

The detection and measurement of stray electrical leakages on dairy farms: effects on performance

Sources of stray electrical leakage from Power Supply Authority alternating current (AC), fence energiser pulses and randomly generated pulses and spikes on 55 dairy farms in the Waikato area were identified between December 1986 and March 1988. The electrical measurements were made using a specifically designed voltmeter able to detect voltages between 0.1V and 1500V, from single voltage spikes of two microseconds or greater duration from direct current (DC) as well as 50 Hz AC. Ninety-five sources of stray voltage were identified, and 53 per cent of properties had more than one source of >0.5V. The major source was from electric fence energisers. Rotary platform parlours were among the commonest sources of random or transient voltage spikes. Leakage of AC into one or other of the components of the milk transport system such as vat, plate cooler, milk lift pump and milk line was common. Owners acknowledged the improvements in milk production, reproductive performance and growth rate of calves after reduction of the exposure of dairy cattle to stray electrical leakage. A representative summary of five case records helps define the range of improvements that may possibly be achieved.     

Histomorphometric analysis of articular cartilage, zone of calcified cartilage, and subchondral bone plate in femoral heads from clinically normal dogs and dogs with moderate or severe osteoarthritis

OBJECTIVE: To quantify and compare the microscopic changes in articular cartilage (AC), zone of calcified cartilage (ZCC), and subchondral bone plate in femoral heads from clinically normal dogs and dogs with moderate or severe osteoarthritis. SAMPLE POPULATION: Femoral heads from clinically normal dogs (n = 16) and dogs with moderate (24) or severe (14) osteoarthritis. PROCEDURES: Femoral heads were allocated to 3 categories (normal, moderate, or severe osteoarthritis) on the basis of radiographic findings, macroscopic findings, and histologic grade determined by use of a modified Mankin scale. Equally spaced 2-mm sections were cut in each femoral head in a coronal or transverse plane. Thickness of the AC, ZCC, and subchondral bone plate was recorded. RESULTS: Mean thickness of AC was significantly greater in samples with moderate and severe osteoarthritis than those considered normal. Mean thickness of the ZCC was significantly greater in samples with moderate and severe osteoarthritis than those considered normal. Mean thickness of the subchondral bone plate in samples with severe osteoarthritis was significantly greater than those with moderate osteoarthritis and those considered normal. A significant decrease in AC thickness was detected in the proximomedial area of femoral heads with severe osteoarthritis, compared with those considered normal. CONCLUSIONS AND CLINICAL RELEVANCE: A cause and effect association between thickening of subchondral structures and thinning and loss of the overlying AC was not detected. Changes in AC were associated with changes in the subchondral bone plate, which is compatible with the theory of adaptation in response to altered load distribution.     

论畜禽产品安全与养殖业的可持续发展

“国以民为本,民以食为天,食以安为先”,食品安全关系到人民身体健康和生命安全,关系到社会稳定和经济发展,关系到对外贸易和国际形象,是具有战略意义的大事。走可持续发展道路,是世界各国政府,特别是生产力发达的国家在新形势下发展本国经济所选择的一个基本原则和发展模式。安全无污染的畜禽产品是健康养殖的重要体规,     

规模种猪场仔猪黄白痢综合防治措施

仔猪黄白痢是由致病性大肠杆菌引起的传染病。根据我县一个规模种猪场的统计,１９９７年１月至１９９８年５月,在死亡仔猪中,因黄白痢引起死亡的占５２％,１９９８年６月份开始对这个种猪场采取综合防治措施,到１９９８年底,共产仔猪３６５０头,２８～３０日龄断奶,成活仔猪３３７２头,成活率达９２３８％,黄白痢发病率为２８％,因黄白痢而死亡的仔猪基本没有发生,成活率比以前有了明显提高,现将综合防治措施报告如下：１　生产母猪的管理和预防接种　生产母猪应加强饲养管理,保证母猪膘情适宜。母猪分娩后立即肌肉注射青霉…     

不同添加剂对晴隆白刺花青贮品质和营养价值的影响

试验通过添加不同种类的青贮剂研究其对晴隆白刺花青贮品质和营养价值的影响.试验设直接青贮、添加乳酸菌（1％FM）、纤维素酶（0.1％FM）、乳酸菌＋纤维素酶（1％ FM ＋0.1％ FM）、甲酸（6 mL/kg FM）、蔗糖（3％FM）及乳酸菌（1％FM）＋纤维素酶（0.1％FM）＋乳酸菌（1％FM）＋纤维素酶（0.1％FM）＋甲酸（6 mL/kg FM）＋蔗糖（3％ FM）7个处理.试验结果表明：青贮过程中pH值总体呈下降趋势,添加剂处理显著低于直接青贮,其中蔗糖的效果最为明显（pH值3.97）,乳酸菌＋纤维素酶＋甲酸＋蔗糖处理也低（pH值4.04）,品质较好;直接青贮的pH值相对较高（4.73）;青贮后晴隆白刺花的粗蛋白（CP）略有下降,乳酸菌＋纤维素酶＋甲酸＋蔗糖降幅最大,达11.18％,但CP含量仍较高,为15.41％,其他各处理CP均有所上升;单独青贮,晴隆白刺花的酸性洗涤纤维（ADF）虽略有增加,但粗纤维（CF）和中性洗涤纤维（NDF）均下降,且NDF降幅较大,达20.15％,而粗脂肪（EE）、无氮浸出物（NFE）含量相对增加,改善了晴隆白刺花的营养;各处理单宁含量均有所下降,其中甲酸处理的降幅最大,达47.83％,乳酸菌＋纤维素酶处理降幅相对较小.     

Comparison of the complement fixation test and competitive ELISA for serodiagnosis of Anaplasma marginale infection in experimentally infected steers

OBJECTIVE: To compare sensitivity of a complement fixation (CF) test and competitive ELISA (cELISA) for detection of Anaplasma marginale in experimentally infected steers. ANIMALS: 40 crossbred (Angus-Simmental) steers. PROCEDURES: Steers were inoculated with 2.6 x 10(9) A marginale-infected erythrocytes (day 0). Blood samples were collected on days 9, 13, 20, 28, 34, 41, 61, 96, 126, and 156 days after inoculation. The percentage of parasitized erythrocytes (PPE) was determined by microscopic examination of stained blood films, and sera were evaluated with the CF test and cELISA by use of USDA-approved methods. Sensitivity and agreement (kappa statistic) between the 2 methods were determined. Persistent infections were confirmed by inoculation of blood obtained from infected steers into susceptible, splenectomized calves. RESULTS: 9 days after inoculation, sensitivity of the cELISA was 47.5%, whereas the CF test failed to identify seropositive steers. After day 13, sensitivity of the cELISA and CF test was 100% and 20%, respectively. During peak parasitemia (day 20), sensitivity of the cELISA and CF test was 100%. Thereafter, sensitivity of the CF test fluctuated between 7.5% and 37.5%, whereas sensitivity of the cELISA remained at 100%. Overall sensitivity of the cELISA and CF test was 94.8% and 26.5%, respectively (kappa statistic, 0.039). CONCLUSIONS AND CLINICAL RELEVANCE: The cELISA had superior sensitivity for serologic detection of A marginale.The CF test and cELISA each had a high percentage of false-negative results during the prepatent period. These findings are relevant for export certification and anaplasmosis prevention or eradication programs.     

Use of computed tomography to evaluate pathologic changes in the lungs of calves with experimentally induced respiratory tract disease

OBJECTIVE: To optimize methods for the use of computed tomography (CT) to assess pathologic changes in the lungs of calves and to determine the effect of treatment on lung consolidation. ANIMALS: 10 male Holstein calves. PROCEDURES: Calves were anesthetized to facilitate CT imaging of the thorax. After initial images were obtained, pneumonia was induced in the calves by inoculation through a bronchoscope. Two calves were used in a preliminary study to refine the inoculation dose and optimize CT images. Four calves were administered florfenicol and 4 calves were untreated control animals. Serial images were obtained 24, 48, and 72 hours after inoculation. After final images were obtained, calves were euthanized, and lung consolidation was estimated by use of lung surface area scoring and water displacement. These estimates were compared with estimated lung consolidation obtained by use of CT. RESULTS: Calves had rapid disease progression. Percentage of lung consolidation was not significantly different between treatment groups for any of the estimation methods. Results of an ANOVA of the 3 assessment methods indicated significant differences among methods. Estimates of the percentage of lung consolidation obtained by use of surface area scoring and CT correlated well, whereas water displacement estimates correlated poorly with other methods of consolidation estimation. CONCLUSIONS AND CLINICAL RELEVANCE: Because of the correlation with other methods for estimation of lung consolidation, CT has the potential to be used to monitor disease progression in calves with experimentally induced respiratory tract disease.     

绵羊包虫病的ELISA诊断试验

在屠宰场通过肝脏观察采集健康的和疑似感染棘球蚴的绵羊血清通过ELISA试剂盒诊断．对ELISA试剂盒的准确性和敏感性进行了评估．发现越是感染后期它的敏感性越高，而在活体中对细粒棘球蚴或多头棘球蚴正确的免疫诊断却比较困难。     

Analytical and Clinical Validation of a Time-resolved Immunofluorometric Assay (TR-IFMA) for Canine C-reactive Protein in Serum

A time-resolved immunofluorometric assay (TR-IFMA) was developed for the determination of C-reactive protein (CRP) in canine serum. CRP was isolated from canine acute-phase serum by affinity chromatography on agarose coupled with phosphorylethanolamine. This isolated dog CRP was used as standard to calibrate the assay. Intra-assay and inter-assay coefficients of variation were in the ranges 5.3–7.1% and 4.8–13.3%, respectively. Accuracy, evaluated by adding 2 and 10 μg/ml of CRP to serum samples, provided recoveries of 99.9% and 106.8%. High correlation was found between CRP measurements by TR-IFMA and a by commercial enzyme-linked immunosorbent assay (R² = 0.98). The limit of detection for the TR-IFMA method was 0.000067 μg/ml and the measurement of CRP in serial dilutions of acute-phase dog sera generated curves with the same slope as the one constructed with purified CRP. The TR-IFMA provides a precise, accurate and highly sensitive assay for CRP determination in dog samples. CRP levels in dogs with different diseases ranged between 10.2 and 210.7 μg/ml and were significantly higher than those observed in healthy dogs (< 7.1 μmg/ml).