<span id="ChDivSummary" name="ChDivSummary" class="abstract-text">为了解牦牛BoLA-I基因mRNA组织表达谱,并探索可能靶定BoLA-I基因的miRNAs,利用RT-PCR技术对牦牛BoLA-I基因mRNA在牦牛肝、脾、肺、肾、肌肉、卵巢、小肠、下颌淋巴结、大肠和肠系膜淋巴结等10种组织的表达谱进行分析,再通过Targetscan和miRBase软件来预测可能靶定BoLA-I基因的miRNAs,检测其在10种牦牛组织中的表达情况。结果表明:BoLA-I基因mRNA在检测的10种牦牛组织中均有表达,尤其在免疫器官组织中广泛表达,在牦牛的肠系膜淋巴结组织表达水平极显著高于小肠和大肠组织（P&lt;0.01）;预测到的可能靶定牦牛BoLA-I基因的miRNA共有28个,从中选择了6个进行组织表达谱分析,发现bta-miR-759、bta-miR-142-5p、bta-miR-665、bta-miR-2322-5p、bta-miR-199a-3p与牦牛BoLA-I基因共表达;除卵巢外,bta-miR-219-5p在其他组织中均有表达,且在肝脏组织中的表达水平极显著高于其他组织（P&lt;0.01）。bta-miR-142-5p在肠系膜淋巴结中表达量极显著高于其他组织（P&lt;0.01）。由此推测,牦牛BoLA-I基因及其预测的miRNA可能在牦牛组织中起重要作用。</span>

旨在探究牦牛Nramp1基因mRNA及可能靶定Nramp1的miRNAs组织表达谱.本研究利用RT-PCR技术对牦牛Nramp1基因的mRNA组织表达谱进行分析,同时运用TargentScan和miRBase软件预测牦牛可能靶定Nramp1基因的miRNAs,并利用加PloyA尾法RT-PCR技术分析miRNAs在肝、脾、肺、肾、后腿骨骼肌、卵巢、小肠、颌下淋巴结、大肠、肠系淋巴结10种组织中的相对表达量.试验结果显示,Nramp1基因mRNA在所检测的10种组织中均有表达,其中在脾、颌下淋巴结及肺组织中的表达量极显著高于肝、肾、大肠、肠系淋巴结组织(P＜0.01),显著高于小肠组织表达量(P＜0.05).预测到可能靶定牦牛Nramp1基因的miRNAs共有201个,从中选择6个进行表达谱分析,发现bta-miR-106a、bta-miR-20b、bta-miR-17-5p在牦牛免疫组织脾、颌下淋巴结、肠系淋巴结中, bta-miR-93、bta-miR-106b、bta-miR-20a在牦牛免疫组织颌下淋巴结、肠系淋巴结中均与Nramp1基因mRNA共表达,但是bta-miR-93、bta-miR-20a、bta-miR-106a、bta-miR-17-5p和bta-miR-20b在颌下淋巴结和肠系淋巴结间的表达量无显著差异(P＞0.05),而bta-miR-106a、bta-miR-20b在颌下淋巴结、肠系淋巴结组织中的表达量极显著高于脾组织表达量(P＜0.01),bta-miR-106b在颌下淋巴结组织中的表达量极显著高于肠系淋巴结组织表达量(P＜0.01).Nramp1基因mRNA在牦牛体内广泛表达说明其可能具有广泛免疫调节作用;bta-miR-93、btamiR-20b、bta-miR-106a、bta-miR-106b、bta-miR-20a和bta-miR-17-5p与Nramp1基因mRNA在免疫组织中的共表达表明二者可能具有靶向调控关系参与免疫调控作用,但二者是否存在真实的靶向调控以及其调控机制有待于深入研究.

Chickens are considered to be potential reservoirs of Newcastle disease virus (NDV). In this study, six Newcastle disease virus strains were isolated and characterized in Tibetan chickens. The HN gene was sequenced, and phylogenetic relationship to reference strains was studied. The phylogenetic analysis demonstrated that these six isolated strains were closely related to NDV isolates of the reference strains GQ245823, KT002186, KU527561, KJ563939, AY225110, EU305607, KM056357, Y18898, GQ245832, AF077761 and lasota strain. Among them, EU305607, KJ563939 and KM056357 were isolated from India, while lasota strain came from attenuated vaccine widely used in China. Then, mean death time (MDT) and intracerebral pathogenicity index (ICPI) were used to estimate the pathogenicity of the isolates. Pathogenicity experiment showed HNH1 and HN17 to be virulent. Our results indicated that genetically diverse viruses circulate in Tibetan chickens, and based upon the phlogeographic analysis, we estimated the origin of ancestral viruses of the isolates and its sister strains located in India and China (lasota strain). It indicates the importance of continuous surveillance to enhance current understanding of the genetic evolution of the NDV strains.

为了探索新城疫病毒(NDV)西藏分离株的免疫效果,以便控制当地NDV的流行,保障禽类健康,试验采用灭活油乳疫苗的制作方法,对藏鸡新城疫病毒(NDV)弱毒株FX10进行了灭活油乳疫苗的研制,并将其免疫效果与商品灭活疫苗进行了比较.结果表明:自制灭活油乳疫苗的各项指标均达到疫苗要求,并能达到商品灭活疫苗的免疫效果.说明NDV弱毒株FX10可作为疫苗候选株.

Newcastle disease (ND) is one of the highly contagious disease responsible for devastating outbreaks and considerable economic losses to poultry industry in China. However, no information is available about NDV in Tibet region; the aim of this study was to determine the genetic characterization and evolutionary analysis of NDV in Tibetan chickens. Four NDVs were isolated from an outbreak in Tibetan chickens. The pathogenicity of the isolates was determined by mean death time (MDT), intracerebral pathogenicity index (ICPI) and amino acid sequencing (112 to 117) of F protein. While the genetic characterization of F gene was determined by sequencing the isolated NDVs and phylogenetic relationship was established with the help of reference strains. Pathogenicity experiments revealed that XZ-F10 and XZ-F20 strains were lentogenic pathotype, while the remaining XZ-F2-1 and XZ-F4-6 strains were velogenic pathotype. The deduced amino acid sequences of the cleavage site of the F protein confirmed our results. Phylogenic analysis of these strains indicated that both XZ-F2-1 and XZ-F4-6 belong to genotype VII. However, XZ-F10 and XZ-F20 strains were assigned to genotypes II. The present study highlights the need for continuous surveillance of NDV in Tibetan chickens; moreover, this study will provide a reference for the local government to make the strategies and control emerging NDV in Tibet.

以《兽医临床诊断学》实验课的现状与存在问题为出发点,在教师的教学自主权范围内,改革模仿式实验教学方式为学生为“主导”的实验教学方式,精心设计实验开设项目,将实验报告式考查改革为实验课程考试等,从而提高了《兽医临床诊断学》的实验教学质量.

Haemophilus parasuis is the causative agent of Glässer's disease, an important emerging infectious disease, but little is known of H. parasuis infection in Tibetan pigs in Tibet. The objective of the present investigation was to examine H. parasuis seroprevalence in Tibetan pigs in Tibet, China. Serum samples from 423 Tibetan pigs in Nyingchi, Tibet, China from April to December in 2010 were examined independently for the presence of antibodies against H. parasuis. A total of 147 (34.75%, 95% confidence interval [CI] 30.21–39.29) Tibetan pigs were tested positive for H. parasuis antibodies by the indirect hemagglutination assay (IHA) using a kit commercially available. 80 of 231 in Nyingchi (34.63%, 95% CI 28.50–40.77) and 67 of 192 in Mainling (34.89%, 95% CI 28.15–41.64) were tested positive, but the difference was not statistically significant (P > 0.05, χ2 = 0.003). The prevalence ranged from 19.72% (95% CI 10.46–28.97) to 75.00% (95% CI 32.57–100) varying in different age groups, with higher prevalence in breeding boars than in piglets, and the difference was statistically significant (P < 0.05). The prevalence of H. parasuis infection in males (45.03%, 95% CI 37.57–52.49) was significantly higher than that in the female (30%, 95% CI 22.41–37.59) pigs (P < 0.05, χ2 = 7.361). Gender of Tibetan pigs was the main risk factor associated with H. parasuis infection. The results of the present survey indicated a wide distribution of H. parasuis among Tibetan pigs in Tibet, China and further investigation should better assess circulation of H. parasuis in Tibetan pigs. To the best of our knowledge, the present study represents the first report of H. parasuis infection in Tibetan pigs in China.

Swine influenza A virus (SIV) is zoonotic pathogen that can be acquired by food-borne transmission because food animals, for example pigs, are recognized as a reservoir. The objectives of this study were to determine the seroprevalence of anti-SIV (H1N1 and H3N2) in Tibetan pigs in Tibet Nationality Autonomous Region, China, a region with cold weather and high altitude. A total of 421 serum samples were randomly collected from Tibetan pigs in Tibet and were evaluated for antibodies against SIV using enzyme-linked immunosorbent assay. Overall, 52 % (219/421) of the animals was positive for H1N1, 16.9 % (71/421) positive for H3N2, and 8.8 % (37/421) positive for both H1N1 and H3N2. The results of the present survey indicated that SIV is highly prevalent among Tibetan pigs in Tibet, China. The results of the present investigation have implications for the ongoing control of SIV infection in Tibetan pigs in Tibet, China and elsewhere.

由于牛病毒性腹泻病毒与猪瘟病毒之间存在共同抗原和交叉免疫,选择A、B两组各15头牛病毒性腹泻病毒检测为阴性的牦牛,分别注射牛病毒性腹泻疫苗和猪瘟疫苗,在不同时间段(第7、14、30、60、90、180天)采集两组牦牛血清,应用酶联免疫吸附试验检测抗体,结果显示两种疫苗都可以使牦牛机体产生抗体,并且牛病毒性腹泻病毒疫苗比猪瘟疫苗整体抗体水平高.

The objective of the present investigation was to determine the seroprevalence of anti-HEV in Tibetan pigs in Tibet Nationality Autonomous Region,China.A total of 421 serum samples were randomly collected from Tibetan pigs in Tibet and were evaluated for antibodies against HEV using enzyme-linked immunosorbent assay(ELISA).204(48.46%) of 421 samples were positive for HEV antibody,and breeding sows had the highest HEV seroprevalence(75%),followed by fattening pigs(60.58%),and piglets had the lowest HEV seroprevalence(31.88%).Generally,there was no statistical difference between male(52.63%) and female(47.89%) Tibetan pigs for the positive rate of anti-HEV(P0.05).Interestingly,the comparison between Nyingchi(53.10%) county and Mainling(43.08%) county showed that the positive rates of anti-HEV in these two regions had significant difference(P=0.04).The results of the present investigation have implications for the ongoing control of HEV infection in Tibetan pigs in Tibet and elsewhere,China.

Chlamydia abortus is an important pathogen in a wide range of animals, associated with abortion in sheep. In this survey, 455 blood samples, collected from Tibetan sheep in Tibet, were examined for antibodies against C. abortus by the indirect hemagglutination (IHA) test. 95 of 455 (20.9%) sheep were seropositive for C. abortus antibodies at the cut-off of 1:16. The seropositivity of C. abortus infection in different age groups varied from 15.2% to 40%, although the differences were no statistically significant (P>0.05). The seroprevalence of C. abortus infection in male sheep (19.4%) was slightly lower than that in females (21.3%), and the difference was not statistically significant (P>0.05). This is the first report of C. abortus seroprevalence in Tibetan sheep in Tibet, China, which has important implications for public health and the local economy.

1 State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, PR China. 2 College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi Province 712100, PR China. 3 College of Animal Science, Tibet College of Agriculture and Animal Husbandry, Nyingchi, Tibet Autonomous Region 860000, PR China.

The seroprevalence of Chlamydiaceae infection in Tibetan pigs in Tibet, China, was examined by indirect hemagglutination assay (IHA), between April, 2010, and December, 2010. A total of 71 of 427 serum samples (16.63%, 95% confidence interval [CI] 15.31-17.95] were positive for Chlamydiaceae antibodies. Forty Chlamydiaceae seropositives from 232 samples were recorded in sera from Nyingchi (17.24%, 95% CI 15.40-19.08) and 31 positives were recorded in 195 serum samples from Mainling (15.90%, 95% CI 14.02-17.78). The investigation showed that the prevalence in female animals was 17.61% (95% CI 15.22-20.00), and in male animals it was 12.72% (95% CI 11.07-14.37). The prevalence ranged from 0% to 20.61% (95% CI 17.81-23.48) among different age groups, with a higher prevalence in growing pigs (p<0.01). The results indicated that Chlamydiaceae infection was widespread in Tibetan pigs in Tibet, China, which is of public health concern in this region of the world. To our knowledge, this is the first report of Chlamydiaceae seroprevalence in Tibetan pigs in Tibet, China.

Toxoplasma gondii infection is widely prevalent in humans and animals, including pigs throughout the world. In this study, the seroprevalence of T. gondii infection in Tibetan pigs in China was investigated for the first time. A total of 427 serum samples were collected from Tibetan pigs in Nyingchi prefecture, Tibet, between April and December 2010, and were assayed for antibodies to T. gondii using the modified agglutination test (MAT). Ninety-seven (22.72%) pigs were found to be positive with MAT titers of 1:25 or higher. Slaughter pigs had the highest seroprevalence, compared with seroprevalence in fattening pigs, growing pigs, or piglets, although the difference was not statistically significant (p≥0.05). The results of the present survey indicate that T. gondii is highly prevalent in Tibetan pigs in Tibet, which poses a significant public health concern in this unique region of the world.

This paper outlined the control of yak major infectious diseases and parasitic diseases, and their prevention and drug therapy, meanwhile, this paper also pointed out that the current lack of control of yak major infectious diseases and parasitic diseases, the development direction for yak disease control was also advised.

The objective of the present investigation was to examine the seroprevalence of Actinobacillus Pleuropneumoniae (APP) in Tibetan pigs in Tibet, China by using an Indirect Hemagglutination Assay (IHA). Serum samples were collected from 332 Tibetan pigs in. Nyingchi, Tibet, China between April and December 2010 and were examined for APP antibodies using a commercially available IHA kit A total of 185 (55.72%) Tibetan pigs were examined seropositive for APP antibodies. The prevalence ranged from 42.68-71.11%, varying in different age groups except breeding boars and sows with higher prevalence in piglets and lower prevalence in slaughter pigs but the difference was not statistically significant (p>0.05). The investigation also showed that the prevalence of APP in female animals was 54.70 and 59.72% in male animals. The results indicated a widespread exposure to APP among Tibetan pigs in. Tibet, Chin. a. This is the first report documenting the infection of APP in. Tibetan pigs in. Tibet, China.

主要对西藏林芝地区某牛场不明原因牛猝死进行研究.通过病死牛病料组织触片镜检和分离培养皆未发现细菌；乳胶凝集试验检查血清为伪狂犬病病毒(PRV)抗体阳性；以脑组织制备DNA模板进行PCR,扩增到特异性的PRV DNA片段；对健康成年兔接种病牛脑、肺等病料,表现典型的伪狂犬病症状.综合流行病学、细菌学、血清学、分子生物学和动物试验结果,确诊为牛伪狂犬病.

取疑似新城疫病毒(NDV)感染的病料进行9日龄鸡胚接种,结果分离到一株病毒。该病毒具有血凝性,血凝活性能被新城疫阳性血清所抑制。减蛋综合征(EDS)阳性血清,传染性支气管炎阳性血清、禽流感H5、H9单因子血清进行血凝抑制(HI)交叉试验发现,该病毒不能抑制血凝活性,动物回归试验出现与新城疫病毒感染鸡相一致的症状,可确认该病为新城疫病毒感染。根据NDV毒力测定标准,MDT>90 h,ICPI为0.38,IVPI值为0,可判定为新城疫弱毒株。

This paper outlined yak major parasitic diseases,its clinical symptoms and drug therapy,meanwhile,this paper also pointed out that the current lack of parasitic diseases control of yak and drug development direction for yak disease parasite control.

取疑似新城疫病毒(NDV)感染的病料进行9日龄鸡胚接种,结果分离到一株病毒.该病毒具有血凝性,血凝活性能被新城疫阳性血清所抑制.减蛋综合征(EDS)阳性血清,传染性支气管炎阳性血清、禽流感H5、H9单因子血清进行血凝抑制(HI)交叉试验发现,该病毒不能抑制血凝活性,动物回归试验出现与新城疫病毒感染鸡相一致的症状,可确认该病为新城疫病毒感染.根据NDV毒力测定标准,MDT＞90 h,ICPI为0.38,IVPI值为0,可判定为新城疫弱毒株.