O型口蹄疫病毒P1-2A3C基因在家蚕杆状病毒表达系统中的表达

口蹄疫是一种严重危害畜牧业生产的烈性传染病。为了促进O型口蹄疫病毒(FMDV)基因工程活载体疫苗的研制,选取O型FMDV编码序列中的衣壳蛋白前体P1-2A基因和蛋白酶3C基因,插入家蚕杆状病毒转移载体pVL1393中,构建重组载体pVL-P1-2A3C,并与线性化病毒Bm-BacPAK6 DNA共转染家蚕BmN细胞,获得重组病毒Bm-P1-2A3C。将重组病毒感染家蚕5龄幼虫,以双抗体夹心ELISA法和间接血凝方法检测血淋巴中的表达产物:目的蛋白在感染病毒后120 h的蚕血淋巴中表达量最高,抗原表达呈阳性的最大稀释倍数为1∶128。结果显示O型FMDV的P1-2A3C基因已在家蚕体内获得表达。     

圆环病毒攻击下生物素添加水平对仔猪细胞免疫及生产性能的影响

将28d断奶、圆环病毒(PVC2)抗体阴性的(杜×长×大)三元杂交仔猪54头,平均分为6组,每组9头,单笼饲养。试验第1天全部猪口鼻接种圆环病毒,在玉米-豆粕型基础日粮中分别添加生物素0、0.10、0.20、0.30、0.40mg/kg和0.50mg/kg,以确定PCV2攻击下,仔猪饲粮生物素添加水平对细胞免疫及生产性能的影响。结果表明:饲粮中添加生物素0.30mg/kg,仔猪外周血E总花环形成率和T淋巴细胞转化率较高,并可促进外周淋巴器官维持正常形态、提高胸腺生长指数;添加0.50mg/kg提高生产性能的效果最佳。由本试验可得出,添加生物素可促进仔猪淋巴器官生长和细胞免疫反应并能提高生产性能,以添加0.30mg/kg的效果较好。     

Evaluation of a new strategy to modulate skeletal development in racehorses by imposing track-based exercise during growth: the effects on 2- and 3-year-old racing careers

REASON FOR PERFORMING STUDY: No data exist on the effects of conditioning exercise at foal age on workload and subsequent clinical injury rate during their 2- and 3-year-old racing careers. OBJECTIVES: To investigate the effects of subjecting TB foals to conditioning exercise prior to the start of race training on: the workload required to reach a level of fitness sufficient to compete; and the prevalence of orthopaedic injury during the first 2 seasons of their racing careers. METHODS: Twenty 18-month-old TBs, 12 subjected to conditioning exercise at foal age (CONDEX) and 8 exercised spontaneously at pasture only (PASTEX) were trained and entered in competitive events. Workload was quantified using the cumulative workload index (the product of average velocity and distance at a specific gait) and the animals were monitored clinically and radiographically for signs of musculoskeletal disorders. RESULTS: Workload to reach the desired fitness level was similar for CONDEX and PASTEX. CONDEX performed more prerace training sessions as 2-year-olds (P<0.05). The incidence of orthopaedic injuries was low in both groups and there were no differences in the occurrence of orthopaedic ailments. PASTEX animals tended to show signs of musculoskeletal disorders earlier than CONDEX animals. This time difference was significant for metacarpophalangeal joint pain on flexion, reduced carpal flexion and hindlimb lameness (P<0.05). CONCLUSIONS: Subjecting TB foals to conditioning exercise early in life does not have adverse effects on racing careers at ages 2 and 3 years, and does not influence the workload needed to reach a fitness level that is sufficient for racing. POTENTIAL RELEVANCE: The lack of negative effects and the indications of some positive effects of early conditioning exercise in the Thoroughbred encourage further large-scale comparative studies.     

Changes in peripheral blood leukocyte subpopulations in piglets co-infected experimentally with porcine reproductive and respiratory syndrome virus and porcine circovirus type 2

Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) are pathogens, which can significantly affect the swine industry worldwide. Field surveys suggest that simultaneous PRRSV and PCV2 infection is common in pigs. The objective of this study was to measure the changes in peripheral blood leukocyte subpopulations in piglets co-infected experimentally with PRRSV and PCV2, in order to analyze the synergistic influence of co-infection on the immune system. Changes in peripheral blood leukocyte subpopulations were systematically measured by flow cytometry (FCM). The levels of antibodies to PRRSV and PCV2 were detected by indirect Enzyme-Linked ImmunoSorbent Assay (ELISA) and the indirect fluorescent antibody test (IFA), respectively. Serum viral loads were measured using real-time PCR. The results showed that piglets co-infected with PRRSV and PCV2 exhibited slower generation and lower levels of antibodies to PRRSV and PCV2, and increased amounts and a prolonged presence of both PRRSV and PCV2 in serum, in comparison to the piglets infected with either virus alone. The major finding in our study was that the total and differential leukocyte counts, including white blood cells (WBCs), monocytes, granulocytes and lymphocytes (T, B and NK cells, as well as T-cell subpopulations), dramatically decreased early during co-infection with PRRSV and PCV2 for about two weeks, in contrast with animals singly infected with either PRRSV or PCV2. These results suggest that PRRSV and PCV2 co-infection results in a synergistic decrease in immune cells in the peripheral blood of piglets. These data contribute to the understanding of the immunosuppressive effects resulting from PRRSV and PCV2 co-infection in pigs.     

紫锥菊对传染性法氏囊疫苗免疫效果的影响

为了探讨紫锥菊作为免疫增强剂的免疫效果和机理,本试验用1日龄的罗斯308肉鸡作为试验材料并用传染性法氏囊疫苗进行免疫。紫锥菊提取物分3种剂量组(A组0.1 mL/只、B组0.5 mL/只、C组1 mL/只)在6日龄开始连续给雏鸡口服7 d,在13,20,27,34,41日龄采血检测外周血中法氏囊病毒抗体,同时测定外周血中IL-2和TNF-α的含量,计算肉鸡的生产性能。结果表明,紫锥菊提取物能明显提高鸡外周血中法氏囊病毒抗体滴度,以及细胞因子IL-2和TNF-α含量,并能改善肉鸡的生产性能。所用3个剂量组中,以B组的0.5 mL/只为最佳的用药剂量。说明紫锥菊提取物具有明显的免疫促进作用。     

不同剂量PMSG对夏季波杂山羊诱导发情效果的影响

采用炔诺酮阴道栓结合孕马血清促性腺激素和人绒毛膜促性腺激素的诱导发情方案对江淮地区波杂山羊进行夏季诱导发情处理,比较了不同剂量孕马血清促性腺激素诱导波杂山羊的发情效果,测定了处理羊外周血液雌二醇和孕酮浓度的变化规律。结果表明,每千克体重肌肉注射5IU和8IU孕马血清促性腺激素时波杂山羊的诱导发情率为100%,显著高于10、12、15IU/kg剂量组的72.22%、71.43%和77.78%,(P<0.05);不同剂量孕马血清促性腺激素诱导发情羊的受胎率差异不显著(P>0.05),孕马血清促性腺激素剂量为8IU/kg时发情羊的受胎率最高(68.75%);处理羊外周血液雌二醇浓度在发情日达到峰值(119.85pg/mL±1.63pg/mL);孕酮在植栓第10天浓度比较高(0.69ng/mL±0.18ng/mL),显著高于对照组的基础水平,撤栓后在发情配种时浓度降低,且低于对照组的基础水平。     

鸡毒支原体不同地区分离株对常用抗菌药物的敏感性试验

从广东、四川和北京等地疑似鸡毒支原体感染鸡中分离并利用种特异性基因（fMG-2）PCR方法鉴定了51株鸡毒支原体,并测定了这些分离株对常用抗菌药物的敏感性。结果表明,三个地区临床分离鸡毒支原体对泰乐菌素仍然保持较好的敏感性,可作为鸡毒支原体感染的首选药物。三个地区的鸡毒支原体对林可霉素和庆大霉素敏感性显著下降,广东、四川两地分离株对强力霉素也有明显耐受性,部分广东分离株对氟喹诺酮类药物敏感性显著下降。     

猪链球菌2型分离株对野生斑马鱼的致病性

以野生斑马鱼为模式动物,对6株mrp＋epf＋sly＋猪链球菌2型（SS2）分离株进行致病性比较。斑马鱼经腹腔接种不同稀释度的SS2分离株,连续观察5d,Kaplan-Meier生存曲线分析并统计半数致死量（LD50）。结果显示,6个分离株对斑马鱼的LD50在10^5cfu-10^6cfu之间,临床株与非临床株的毒力差异不显著（P〉0.05）。从攻毒后死亡的鱼腹腔和脑可分离到SS2,并伴有腹腔出血及肝、肠、脑部炎性病变。表明SS2可感染斑马鱼,为进一步研究SS2的体内感染机制及毒力因子功能等奠定了基础。     

犬新孢子虫NcSRS2基因原核表达质粒的构建

根据已发表的犬新孢子虫NcSRS2基因序列,设计1对含有EcoRⅠ和NotⅠ酶切位点的引物。以提取犬新孢子虫虫体基因组DNA为模板,应用PCR扩增获得NcSRS2 ORF基因片段,将此基因片段克隆到pMD18-T Simple载体上,用EcoRⅠ和NotⅠ双酶切该片段,回收得到含有两个酶切位点黏端的Nc-SRS2 ORF基因,将此基因片段克隆至相同酶切回收后的PGEM-4T-2原核表达载体中,获得重组质粒pGEX-NcSRS2,经PCR鉴定,限制性内切酶分析和克隆片段序列测定比较,证实了重组质粒的正确性。     

鸡传染性法氏囊病病毒RT-PCR检测方法的建立

根据GenBank中已经发表的传染性法氏囊病病毒（IBDV）VP2基因的高度保守序列,设计并合成了一对引物,Blast在线检索GenBank数据库,未发现其他相似序列。提取已鉴定的IBDV—QD株传代病毒尿囊液的总RNA,合成cDNA模板,优化RT-PCR反应条件,最终获得预期453bp的目的片段,IBDV扩增产物经测序结果证实与GenBank中已发表的致病力不同的IBDV毒株相应序列同源性达97．4％～99．9％,最终建立了RT—PCR检测方法。用已建立的RT—PCR方法对已知的8份临床IBDV阳性法氏囊病料进行检测,阳性率达100％,另外对2份鸡白血病病毒（ALDV）,2份鸡传染性喉气管炎病毒（Infectious laryngotracheitis virus,ILTV）,2份鸡传染性支气管炎病毒（IBV）阳性病料进行平行同条件检测,结果均为阴性。表明所建立的RT—PCR特异性好、敏感性高,可用于鸡传染性法氏囊病的临床初步诊断及流行病学调查。