禽腺病毒血清4型感染鸡组织中NLRP3基因转录水平的实时荧光定量PCR分析

旨在分析禽腺病毒血清4型（FAdV-4）感染鸡组织中NLRP3基因的转录水平,本研究设计鸡NLRP3特异性引物,利用RT-PCR扩增NLRP3基因180 bp片段并克隆至pMD-18T载体,制备重组质粒pMD-18T-NLRP3。以pMD-18T-NLRP3质粒作为标准品进行荧光定量PCR并建立标准曲线。通过反应条件优化,成功建立了检测NLRP3基因的实时荧光定量PCR方法,并利用该方法对致病性FAdV-4感染鸡组织中NLRP3基因的转录水平进行了分析。结果显示,所设计的NLRP3引物可特异性扩增鸡NLRP3基因,建立的实时荧光定量PCR对鸡NLRP3标准质粒的扩增曲线良好,标准品的拷贝数与Cq值呈现良好的线性关系。与对照组相比,NLRP3分子在FAdV-4感染鸡肝和脾中的转录水平极显著高于对照组（P<0.001）,在盲肠扁桃体和法氏囊的表达显著高于对照组（P<0.01）。本研究所建立的鸡NLRP3基因SYBR Green Ⅰ实时荧光定量PCR可以检测FAdV-4感染鸡不同组织中NLRP3的转录水平;致病性FAdV-4感染所造成的组织炎症损伤与NLRP3分子密切相关。     

Molecular characterisation of Theileria equi and risk factors associated with the occurrence of theileriosis in horses of Punjab (Pakistan)

Theileria equi (T. equi) is an obligate intra- and extra-erythrocytic parasite that causes equine theileriosis (ET) in equids. Equine theileriosis is considered a notifiable disease of global significance, a major constraint to the international movement of horses, and endemic in many countries. This disease may be difficult to diagnose, as it can produce variable and nonspecific clinical signs. A cross-sectional study was designed for the molecular characterisation of T. equi and to investigate the associated risk factors of ET accompanied by its consequences on haematological and sero-biochemical parameters. A convenience sampling of 500 blood samples were collected from ET suspect horses from January to December 2017. PCR was performed on all blood samples targeting the 18S rRNA gene of T. equi followed by sequencing; 9% animals tested positive with confirmed sequences. The isolates of this study showed high homology with Cuban, Russian and Brazilian isolates of T. equi (accession numbers KY111762.2 , MG551915.1 and KY952237.1 , respectively). Based on multivariate analysis, the principal risk factors consisted of absence of dogs on the premises and presence of tick infestation. The haemato-biochemical parameters showed a decrease in granulocytes and erythrocytes, and an increase in lymphocytes, monocytes, mean corpuscular volume, mean corpuscular haemoglobin, mean platelet volume, glucose, phosphorus and aspartate aminotransferase in positive horses. This is the first study which identified ET in Punjab (Pakistan) using molecular techniques and risk factors together with the haemato-biochemical variations in horses.     

我国羊隐孢子虫感染情况及危险因素

隐孢子虫是可以感染小反刍动物的肠道原生生物,具有潜在的公共卫生学问题。隐孢子虫可引起人和动物的腹泻和肠道疾病,严重情况下会出现死亡。被感染的动物可能是人畜共患型隐孢子虫的宿主,可引起公共卫生风险、农场利润减少和动物福利等问题。隐孢子虫病已被报道是新生牛、羊等反刍动物腹泻和死亡的一个重要原因,认为是新生羔羊腹泻的第二大诱因,仅次于轮状病毒。羊隐孢子虫病在世界范围内都有发现,不同程度地威胁着人和动物的身体健康,基于此,现就隐孢子虫对羊感染的危险因素和常用检测方法,以及我国羊感染的现状进行综述,为其防控提供参考。     

AANAT基因在滩羊不同繁殖时期卵巢中的转录差异及DNA甲基化程度分析

试验旨在检测5-羟色胺-N-乙酰基转移酶（AANAT）基因在绵羊休情季节和繁殖季节（卵泡期和黄体期）卵巢组织中的转录差异,并分析转录差异是否由DNA甲基化修饰程度改变所导致。试验采用自然环境条件和饲养管理一致,且体重差异在0.5 kg范围内的空怀母滩羊作为试验动物,采集其休情期、卵泡期和黄体期（每个时期3只）的卵巢组织,采用SYBR染料法进行实时荧光定量PCR检测AANAT基因在滩羊不同繁殖时期卵巢组织中的转录水平。随后针对转录水平有差异的两个时期（休情期和卵泡期）的样本,利用MethPrimer 2.0在线软件预测AANAT基因启动子区和第一外显子区的CpG岛;用重亚硫酸盐测序法（BSP法）检测AANAT基因启动子区及第一外显子区的甲基化程度。试验结果显示,滩羊休情期卵巢组织中AANAT基因转录水平显著低于卵泡期的AANAT基因转录水平（P<0.05）,休情期与黄体期滩羊卵巢组织中AANAT基因的转录水平差异不显著（P>0.05）。滩羊卵巢组织中AANAT基因启动子区上存在着一个长度为173 bp的CpG岛,第一外显子区存在着一个长度为118 bp的CG岛。然而,两个甲基化岛区内的单个CpG位点甲基化程度在滩羊休情期和卵泡期之间均不存在显著差异,暗示AANAT基因的表达受甲基化修饰外的因素调控。本研究结果可为进一步探讨AANAT基因在季节性发情和卵泡成熟中的功能提供参考资料。     

Avian anaesthesia related mortality and the associated risk factors in a UK zoological collection

ObjectiveTo analyse avian anaesthesia-related mortality in a UK zoological collection over a 5-year period and identify risk factors for mortality.Study designRetrospective cohort study.AnimalsA total of 135 individual birds across 37 species, anaesthetized during 206 events in a UK zoological collection between 1 January 2014 and 30 June 2019 (inclusive).MethodsAnaesthesia records were reviewed and variables such as age, body condition, weight, sex, duration of anaesthesia and health status were collated. Anaesthesia-related mortality was defined as those deaths occurring during anaesthesia and up to 7 days postanaesthesia. Outcome was analysed using multivariable conditional logistic regression. Overall mortality was defined and included birds euthanised during anaesthesia for non-anaesthesia related reasons. Data were summarised as median (range). A value of p < 0.05 was considered significant. Relative risks and 95% confidence intervals (95% CI) were calculated for the association between risk factors and anaesthetic death where a statistically significant difference was found.ResultsThe overall mortality rate was 10.19% (95% CI = 6.06–14.3%), while anaesthesia-related mortality was 3.88% (95% CI = 1.69–7.51%). Birds with an abnormal health status had a 15.53-fold (95% CI = 1.95–123.63) increased risk of death compared with those with a normal health status. The duration of anaesthesia was also a statistically significant risk factor (p = 0.021) in the univariable analysis, but not when combined with health status. No other variables were associated with anaesthesia-related mortality.Conclusions and clinical relevanceAbnormal health status and longer anaesthetic procedures were associated with a significantly increased risk of anaesthesia-related death in this population of birds. It is recommended that anaesthetic duration is minimized, and pre-existing diseases are diagnosed where possible prior to general anaesthesia of birds. Anaesthetizing healthy birds was associated with a low risk of mortality.     

有关土地测绘质量问题和处理研究

文章就土地测绘的内涵解析作为切入点,浅要论述了土地测绘质量问题的影响因素,并就土地测绘质量问题影响因素的处理提出了一些浅薄的建议。以期促进我国土地测绘质量的提升。     

地温空调井抽回灌效果影响关键因子识别

为了探析引起地温空调井抽回灌效果的主要制约因素,通过单相关系数分析、主成分绝对得分的源解析模型、拟合度检验和多元线性回归的方法分析影响单井回灌效率的影响因素。结果表明:井径、滤水管孔隙率及抽回灌井数比与单井回灌效率的相关性和拟合度较好,故将井径、滤水管孔隙率及抽回灌井数比作为影响单井回灌效率的关键因子,同时分析了单井回灌效率与关键因子的之间的拟合方程及多元线性回归方程,得到了单井回灌效率与关键因子之间的定量关系。     

怀孕期高原鼢鼠生物学特征与环境因子的关系分析

选择甘肃天祝县高原鼢鼠(Mospalax baileyi)2个地理位置不同的栖息地为研究区,以高原鼢鼠体重、胎数、妊娠率和种群密度作为其生物学特征指标,以2011~2013年归一化植被指数(NDVI)、年均降水量和年均温度为环境因子,分析二者的相关关系。结果表明:2个研究区域NDVI存在显著性差异(P0.05);除了2011年和2013年雌性体重存在差异外,2个区域高原鼢鼠体重、妊娠率、平均胎数和种群密度均无显著性差异(P0.05);雌雄个体重和平均胎数与NDVI呈负相关关系,妊娠率和种群密度与NDVI呈正相关关系;雄性个体重、胎数和种群密度与年均温呈负相关关系,雌性个体重和妊娠率与年均温呈正相关关系;雄性个体重、妊娠率、胎数和种群密度与年均降水量呈正相关关系,雌性个体重与年均降水量呈负相关关系,但高原鼢鼠各生物学指标与环境因子相关性均不显著(P0.05)。说明,高原鼢鼠体重、妊娠率、胎数和种群密度的变化与栖息地环境因子没有直接关系,生物学特征受环境因子的影响较小。     

Apparent seroprevalence,isolation and identification of risk factors for brucellosis among dairy cattle in Goa,India

Brucellosis is a highly contagious zoonotic infection affecting livestock and human beings. The disease has been reported worldwide except in few countries where it has been eradicated. The prevalence of brucellosis among cattle from 11 farms having a history of abortions was studied. A total of 481 samples comprising of blood, milk, vaginal swabs, vaginal discharges, placental tissues and fetal tissues were collected from 296 animals. Clinical samples were processed for the isolation of Brucella. Serum samples (n = 296) were tested by Rose Bengal Plate Test (RBPT) and indirect ELISA. A total of 90 (30.40%) and 123 (41.55%) samples were positive by RBPT and indirect ELISA, respectively. Also 27.02% samples were positive by both the tests. Brucella isolates (n = 8) were recovered from clinical samples using Brucella selective media. All the isolates demonstrated PCR amplification for the bcsp31 and IS711 genes. Amplification of Brucella abortus specific primer was demonstrated by all the isolates in AMOS PCR indicating isolates to be of either B. abortus biotype 1, 2 or 4. Risk factors for transmission of brucellosis among cattle population were studied by field surveys. It was observed that lack of awareness about brucellosis (OR = 8.739, P = 0.138) and inadequate floor space (OR = 0.278, P = 0.128) were crucial risk factors for transmission of bovine brucellosis.