捻转血矛线虫离体培养的研究

本实验研究了:(1)三种人工合成培养基:M199、NCTC109、F_(12);(2)小牛脱纤维蛋白血(以下简称小牛脱纤血)、Filde’s试剂,Bovine hemin;(3)pH值等几种因素对离体培养中捻转血矛线虫(Haemonchou contortus)L_3发育至L_4的影响。结果表明:(1)在相同条件下,NCTC109和M199培养系中的L_3可发育至L_4晚期,F_(12)中L_3只发育至L_4早期;三种培养系中L_3发育至L_4的虫体数占存活虫体数的90％以上,以NCTC109中的发育率为最高,达100％。(2)小牛脱纤血、Filde’s试剂和Bovine hemin对L_3发育至L_4均有促进作用,虫体的存活率明显提高,晚四期幼虫的存活时间延长;小牛脱纤血和Filde’s试剂可使L_3至晚L_4的时间缩短。(3)pH 5.8～6.2时,L_3发育至L_4;速度明显减慢,发育率不到50％;pH 6.6～6.8时,L_3发育至L_4的同步性及发育率明显提高,以pH6.6为最佳。     

Fatal meningitis in a calf caused by Mannheimia varigena

Mannheimia varigena was identified as the etiologic agent of meningitis in a young Belgian White Blue heifer calf. Species identification of the bacterium was done by phenotyping and molecularly confirmed by tDNA-PCR. Standard bacteriological examination might fail to differentiate species belonging to the genus Mannheimia.     

Nuclear receptor and target gene mRNA abundance in duodenum and colon of dogs with chronic enteropathies

Nuclear receptors (NR), such as constitutive androstane receptor (CAR), pregnane X receptor (PXR) and peroxisome proliferator-associated receptors alpha and gamma (PPAR, PPARγ) are mediators of inflammation and may be involved in inflammatory bowel disease (IBD) and food responsive diarrhea (FRD) of dogs. The present study compared mRNA abundance of NR and NR target genes [multi drug-resistance gene-1 (MDR1), multiple drug-resistance-associated proteins (MRD2, MRD3), cytochrome P450 (CYP3A12), phenol-sulfating phenol sulfotransferase (SULT1A1) and glutathione-S-transferase (GST A3-3)] in biopsies obtained from duodenum and colon of dogs with IBD and FRD and healthy control dogs (CON; n = 7 per group). Upon first presentation of dogs, mRNA levels of PPAR, PPARγ, CAR, PXR and RXR in duodenum as well as PPARγ, CAR, PXR and RXR in colon were not different among groups (P > 0.10). Although mRNA abundance of PPAR in colon of dogs with FRD was similar in both IBD and CON (P > 0.10), PPAR mRNA abundance was higher in IBD than CON (P < 0.05). Levels of mRNA of MDR1 in duodenum were higher in FRD than IBD (P < 0.05) or CON (P < 0.001). Compared with CON, abundances of mRNA for MRP2, CYP3A12 and SULT1A1 were higher in both FRD and IBD than CON (P < 0.05). Differences in mRNA levels of PPAR and MRP2 in colon and MDR1, MRP2, CYP3A12 and SULT1A1 in duodenum may be indicative for enteropathy in FRD and (or) IBD dogs relative to healthy dogs. More importantly, increased expression of MDR1 in FRD relative to IBD in duodenum may be a useful diagnostic marker to distinguish dogs with FRD from dogs with IBD.     

河南省鲁山县规模养猪场仔猪腹泻调查报告

通过对5家养猪场仔猪腹泻调查,了解鲁山县仔猪腹泻流行现状,以便为今后更好地做好防控工作提供参考依据.     

牛病毒性腹泻病毒与宿主细胞的相互作用研究

自1946年Olafson等首次在美国纽约州发现牛病毒性腹泻病毒(BVDV)以来,该病毒在世界各地广泛流行和传播,尤其对一些畜牧业发达国家的奶业和牛肉业造成了巨大损失.了解牛病毒性腹泻病的致病机理,研制出更加安全可靠有效的疫苗,对控制该病的发生和蔓延尤为重要.对BVDV的病原学、生物学特性以及对宿主细胞的相互作用等进行了阐述,以期为BVDV的预防及治疗奠定基础.     

发酵黄芪对断奶仔猪日增重、腹泻率及免疫能力的影响

将发酵黄芪添加于断奶仔猪基础日粮中,与基础日粮及抗生素作比较,研究其对断奶仔猪日增重、腹泻及免疫功能的影响。结果表明,与基础日粮组相比,发酵黄芪能显著提高仔猪日增重、降低腹泻率及免疫能力;与抗生素组相比,发酵黄芪能显著降低仔猪腹泻率,提高免疫能力。为生产新型功能饲料提供良好途径,为中药饲料添加剂替代抗生素研究奠定基础。     

猪源牛病毒性腹泻病毒套式PCR检测方法的建立

根据GenBank中BVDV-1、BVDV-2、CSFV及新出现瘟病毒的基因序列,设计2对特异引物,建立了能鉴别诊断BVDV与CSFV的Nested-PCR检测方法;确定其方法的特异性、敏感性、稳定性。建立的Nested-PCR能从BVDV标准毒株、猪源BVDV毒株中扩增198bp特异性片段,对猪瘟病毒、猪繁殖与呼吸综合征病毒检测结果为阴性。结果表明,该方法具有较好特异性;其敏感性可检测到0.195fg RNA模板量;经重复性试验表明该方法具有良好稳定性。初步应用检测表明,猪BVDV阳性率较高,达36.0%;牛血清及其制品、猪瘟活疫苗BVDV污染严重。本试验建立的Nested-PCR具有敏感、特异和稳定等特点,可用于临床诊断和流行病学调查。     

1株猪源牛病毒性腹泻病毒的分离与鉴定

从BVDV阳性仔猪病料中分离病毒,为开展猪源BVDV病原学研究奠定基础。将处理后BVDV阳性仔猪组织样品接种MDBK细胞,分离到1株猪源BVDV,命名为SD0803株。通过细胞培养、直接免疫荧光、5′-UTR与Npro PCR扩增、电镜观察、TCID50测定及对其分子进化特征加以分析。结果表明,该毒株在MDBK细胞上盲传至13代未出现细胞病变。在直接免疫荧光试验中呈阳性荧光信号。PCR扩增分别获得5′-UTR与Npro预期大小DNA片段。电镜观察,病毒粒子略呈圆形,有囊膜,直径约50nm。病毒滴度为10-6.5 TCID50.0.2 mL-1。SD0803 5′-UTR、Npro序列进化分析显示,该分离株属于BVDV-1,与已知BVDV-1各亚型之间同源性较低,单独成一分支。结果表明,成功分离鉴定1株猪源BVDV SD0803,该毒株为非致病变型BVDV-1,极有可能为BVDV-1新的亚型。     

猪流行性腹泻病毒ORF3和M基因的克隆与序列分析

为了解福建省猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)毒株ORF3和M基因变异情况,本研究从4个不同规模猪场发生仔猪“顽固性腹泻”疾病的病料中扩增到4株PEDV的ORF3和M基因,并进行了序列分析.结果表明:4株PEDV ORF3基因均含有675个碱基,编码224个氨基酸,与GenBank中登录的代表性PEDV核苷酸的同源性为89.7％～100.0％,氨基酸的同源性为94.7％～99.6％,其中FJPT毒株核苷酸的同源性与CV777 truncated毒株最低,仅为89.7％,FJNP毒株核苷酸的同源性与中国北方流行的CH ZKFG 11毒株最高,达100.0％.M基因含有681个碱基,编码226个氨基酸,与GenBank中登录的代表性PEDV核苷酸的同源性为94.8％～100.0％,氨基酸的同源性为94.8％～99.6％,其中FJPT毒株核苷酸的同源性与日本D89752毒株最低,仅为94.8％,FJNP毒株核苷酸的同源性与韩国CPF193、泰国毒株最高,达100.0％.4株PEDV毒株均与国内外流行强毒株的亲缘关系较近,与attenuate DR13弱毒株的亲缘关系较远.