猪猝死症4种重要病原多重连接探针扩增鉴别检测技术的建立与应用

为了快速检测引起猪猝死的病原,本研究针对临床引起猪猝死症的4种疾病病原(猪尼帕病毒、非洲猪瘟病毒、猪产气荚膜梭菌和猪胸膜肺炎放线杆菌)分别设计特异性探针,建立了同时检测这4种病原的多重连接探针扩增技术(MLPA)。对扩增产物进行毛细管电泳分析,结果显示该方法,探针之间没有交叉反应,特异性强。对不同病原核酸的最低检测限可以达到140拷贝μ/L～370拷贝μ/L,敏感性较高。应用该MLPA方法对129份临床样品进行检测,并与国家相关标准进行比较。结果显示,两种方法的符合率为100%,且该方法检测猪产气荚膜梭菌和猪胸膜肺炎放线杆菌的特异性和敏感性均达到100%,Kappa值均为1。该方法的检测结果与国家或行业标准PCR的检测结果一致性较高,且克服了后者无法同时进行多重检测的缺点,最多可以同时检测45种病原,对于复杂症候群的快速和高通量检测具有重要临床意义。该方法可推广到其它多重病原体的检测中,显示出广阔的应用前景。     

上海市崇明区生猪屠宰行业及监管现状调查

为了解辖区内生猪屠宰行业现状,进一步完善生猪屠宰监督管理,上海市对现存的2家生猪定点屠宰企业及检疫队伍开展了调查研究。结果显示:本区生猪定点屠宰的猪源主要为崇明本地猪,屠宰检疫合格率达99.99%,斥品检出率低于0.5%的警戒线,产品主要销往上海市;屠宰检疫队伍由10名官方兽医和14名签约兽医组成,官方兽医具有年轻化、高学历等特征,签约兽医年龄阶层分布合理,具有从业经验丰富等特征;企业"瘦肉精"抽检率高于3%的最低标准,非洲猪瘟自检达到"批批检、全覆盖"的要求。调研中发现存在生猪产品供需不平衡,屠宰检疫队伍综合水平不高,屠宰企业自检费用高等问题,这提示应发展生猪产加销全产业链,开展检疫人员分类培训并改善待遇,积极争取政策资金支持,推动生猪屠宰监管的高质量发展。     

基于ISSR标记初选格鲁桑类型桑树核心种质

核心种质可以用最少份数的种质资源代表该物种及生态类型的遗传多样性。以收集自我国黄土高原的73份格鲁桑类型桑树种质资源为材料,利用15个ISSR引物共扩增出129条带,其中多样性条带为115条,多态性比率为89.15%。根据ISSR分子标记聚类结果和树型图,利用逐步聚类随机取样法和属性约简启发式算法对73份种质资源进行核心种质构建研究,2种方法分别初选出21份和16份格鲁桑类型桑树核心种质,核心种质样本的比率分别为28.77%、21.92%。利用统计软件SPSS 13.0,结合分子标记多态性位点数、多态性位点百分率、观测等位基因数、有效等位基因数、Nei's遗传多样性指数和Shannon's信息指数对2组核心种质的遗传多样性进行评价和t检验,结果显示:采用2种方法初选出的21份和16份格鲁桑核心种质都能很好地保存初始群体的遗传多样性和结构,但采用属性约简启发式算法所得16份核心种质的代表性要优于采用逐步聚类随机取样法所得的21份核心种质。最终确定以属性约简启发式算法初选的16份样本作为格鲁桑类型桑树核心种质。     

猪圆环病毒与胸膜肺炎放线杆菌混合感染的诊治

2019年3月,大庆某规模化猪场个别猪出现发育不良、咳喘、消瘦、皮肤苍白和呼吸困难等症状,随后出现死亡。为明确发病原因,降低损失,对送检病死猪进行临床症状观察、剖检及实验室检测,并对病原菌进行药物敏感性试验。检测结果为致病性胸膜肺炎放线杆菌和猪圆环病毒混合感染所致,分离的胸膜肺炎放线杆菌对阿莫西林和头孢唑高度敏感。本研究为养殖场饲养猪的死亡病因分析和防治提供了参考。     

黔育1号菊苣与紫花苜蓿、鸭茅间作栽培效果研究

依据贵州省松桃县的气候特点和牧草种植需要,用黔育1号菊苣与北林202紫花苜蓿、黔草4号鸭茅分别进行间作试验,从产量、营养品质、生产效益等方面对间作复合群体与3种牧草单播进行比较分析,探讨黔育1号菊苣与不同牧草间作种植模式的增产效益。结果表明,在贵州松桃地区,黔育1号菊苣+北林202紫花苜蓿间作,年均鲜草产量可达87.27 t/hm²,比紫花苜蓿单播产量增加36.17%,生产效益提高20.77%,粗蛋白含量达26.83%。该研究结果可为松桃县种草养畜产业的发展提供技术支撑。     

不同水平的植物乳杆菌代谢产物对肉鸡生长性能、粪微生物菌群及绒毛形态的影响

本试验在日粮中添加不同水平的植物乳杆菌代谢产物,研究其对肉鸡生长性能、粪微生物菌群、挥发性脂肪酸含量及小肠绒毛形态的影响。选择健康、体重一致的1日龄AA肉鸡576只,采用单因素试验设计,随机分为6组,每组4个重复,每个重复24只鸡,对照组饲喂基础日粮+80 mg/kg新霉素,处理组分别在基础日粮中添加0.1%、0.2%、0.3%、0.4和0.5%不同水平的植物乳杆菌代谢产物。试验共进行42 d。结果显示:0.4%植物乳杆菌代谢产物组较对照组、0.1%、0.3%和0.5%植物乳杆菌代谢产物组显著提高了42 d肉鸡末重、总增重和平均日增重(P <0.05),0.1%和0.3%植物乳杆菌代谢产物组较0.4%植物乳杆菌代谢产物组显著提高了肉鸡饲料转化率(P <0.05)。对照组较各植物乳杆菌代谢产物组显著提高了粪中大肠杆菌含量(P <0.05),与0.4%植物乳杆菌代谢产物组相比,对照组、0.1%、0.2%和0.5植物乳杆菌代谢产物组显著降低了粪中乙酸含量(P <0.05),0.1%植物乳杆菌代谢产物组较0.3%、0.4%和0.5%植物乳杆菌代谢产物组显著提高了粪中丁酸含量(P <0.05)。0.4%植物乳杆菌代谢产物组总挥发性脂肪酸含量最高,显著高于对照组(P <0.05)。0.4%植物乳杆菌代谢产物组十二指肠和空肠绒毛高度显著高于其他各组(P <0.05),0.2%和0.3%植物乳杆菌代谢产物组十二指肠绒毛高度显著高于对照组、0.1%和0.5%植物乳杆菌代谢产物组(P <0.05),0.2%植物乳杆菌代谢产物组空肠绒毛高度显著高于对照组、0.1%和0.5%植物乳杆菌代谢产物组(P <0.05)。0.1%植物乳杆菌代谢产物组回肠绒毛高度最高(P <0.05)。对照组和0.1%组植物乳杆菌代谢产物十二指肠和空肠隐窝深度显著高于0.4%和0.5%植物乳杆菌代谢产物组(P <0.05),但对照组回肠隐窝深度最低(P <0.05)。结论 :日粮添加植物乳杆菌代谢产物可以改善肉鸡生长性能,降低粪中大肠杆菌含量,提高小肠绒毛高度和粪中挥发性脂肪酸含量,其中日粮添加0.2%和0.4%植物乳杆菌代谢产物对肉鸡增重、饲料转化率和绒毛高度的影响最佳。     

Baculovirus virions displaying infectious bursal disease virus VP2 protein protect chickens against infectious bursal disease virus infection

Infectious bursal disease (IBD) is an acute and contagious viral infection of young chickens caused by IBD virus (IBDV). The VP2 protein of IBDV is the only antigen for inducing neutralizing antibodies and protective immunity in the natural host. In the current study, we have succeeded in construction of one recombinant baculovirus BacSC-VP2 expressing His6-tagged VP2 with the baculovirus envelope protein gp64 transmembrane domain (TM) and cytoplasmic domain (CTD). The His6-tagged recombinant VP2 was expressed and anchored on the plasma membrane of Sf-9 cells, as examined by western blot and confocal microscopy. Immunogold electron microscopy demonstrated that the VP2 protein of IBDV was successfully displayed on the viral surface. Vaccination of chickens with the VP2-pseudotyped baculovirus vaccine (BacSC-VP2) elicited significantly higher levels of VP2-specific enzyme-linked immunosorbent assay antibodies and neutralizing antibodies than the control groups. IBDV-specific proliferation of lymphocytes was observed in chickens immunized with the recombinant BacSC-VP2. An in vivo challenge study of the recombinant baculovirus BacSC-VP2 showed effective protection against a very virulent (vv) IBDV infection in chickens. In addition, mortality and gross and histopathological findings in the bursa demonstrated the efficacy of the vaccine in reducing virulence of the disease. These results indicate that the recombinant baculovirus BacSC-VP2 can be a potential vaccine against IBDV infections.     

Immunoadjuvant activities of a recombinant chicken IL-12 in chickens vaccinated with Newcastle disease virus recombinant HN protein

Recombinant fowlpox virus (rFPV/HN) expressing Newcastle disease virus (NDV) HN gene and rFPV/HN/chIL-12 co-expressing chicken IL-12 (chIL-12) and HN (rHN/chIL-12) genes have been characterized. rHN/chIL-12 or rchIL-12, expressed by our previous construct rFPV/chIL-12, co-administered with rHN was assessed for adjuvant activities of chIL-12. Chickens were vaccinated with various amounts of rHN/chIL-12 mixed with mineral oil (MO), intramuscularly. Levels of hemagglutination-inhibition (HI) antibody production depended on the concentration of the injected rHN or rHN/chIL-12. The lower HI antibody titers were obtained in chicken groups rHN/chIL-12/7-rHN/chIL-12/9, receiving 60ng rHN/8ng chIL-12 with MO, 30ng rHN/4ng chIL-12 with MO or 15ng rHN/2ng chIL-12 with MO, respectively, compared to those in chicken groups rHN/7-rHN/9, receiving rHN with MO alone. However, chickens in group rHN/chIL-12/7 or rHN/chIL-12/8 and rHN with MO alone showed the same effective protection. Chicken group rHN/chIL-12/9 was even more protective than that in group rHN/9. When rchIL-12 was co-injected with 15ng rHN plus MO, chickens produced low levels of HI antibody titers; while higher levels of IFN-γ production and an effective protection rate (83%) were obtained. On the other hand, low levels of IFN-γ production and low protection response (50%) were obtained in chickens injected with rHN with MO alone. Taken together, when the concentration of rHN decreased to certain levels, rchIL-12 reduced HI antibody production. The increase in the induction of IFN-γ production might suggest the enhancement of the cell-mediated immunity which conferred the protection from the NDV challenge.     

饲粮中添加牛至精油对绵羊复胃发育、消化酶活性及瘤胃微生物区系的影响

本试验旨在研究饲粮中添加牛至精油对绵羊复胃发育、消化酶活性及瘤胃微生物区系的影响。选取18只体重相近[(20.30±1.27) kg]、体况良好的3月龄萨福克×小尾寒羊F1代公羔,随机分为3组(每组6只):对照组(CON组)饲喂基础饲粮,牛至精油组饲喂在基础饲粮基础上分别添加4(EO4组)和7 g/d(EO7组)牛至精油的试验饲粮,预试期10 d,正试期72 d。结果显示:1)绵羊瘤胃容积随饲粮中牛至精油添加量的增加显著增大(P<0.05),与CON组相比,EO7组瘤胃容积指数提高了2. 09%(P <0. 05),皱胃容积指数降低了1. 61%(P <0. 05)。2)与CON组相比,EO4组瘤胃中纤维素酶和胃蛋白酶活性显著升高(P<0.05),皱胃中纤维素酶和β-葡萄糖苷酶活性显著升高(P<0.05); EO7组瘤胃、网胃中纤维素酶、β-葡萄糖苷酶、α-淀粉酶活性显著升高(P<0.05),瓣胃中纤维素酶和皱胃中胃蛋白酶活性显著升高(P<0.05)。3)饲粮添加4和7 g/d牛至精油均显著降低了瘤胃中原虫数量(P<0.05);与CON组相比,EO4组瘤胃中黄色瘤胃球菌数量显著升高(P<0.05); EO7组瘤胃中真菌、黄色瘤胃球菌、白色瘤胃球菌和产琥珀酸丝状杆菌数量均显著升高(P<0.05)。综上所述,饲粮中添加牛至精油能促进瘤胃发育及瘤胃中真菌和纤维分解菌的增殖,抑制原虫的生长,提高瘤胃消化酶活性,且以7 g/d的添加量效果较好。     

过表达NR4A1促进山羊皮下脂肪细胞分化

旨在克隆山羊N R4A1基因的CDS区序列,明确其组织和细胞表达模式,以及探究过表达N R4A1基因对山羊皮下脂肪细胞分化的影响.本试验利用双酶切法构建山羊过表达载体pcDNA3.1-NR4A1.以1周岁简州大耳羊(n=5)为试验动物.利用RT-PCR方法和实时荧光定量PCR(real-time quantitativ...