应用核酸斑点杂交法检测鹅细小病毒(GPV)

利用鹅细小病毒 (GPV)核酸探针 ,设计并建立了核酸斑点杂交法 ,用以检测 GPV核酸。该方法特异性与敏感性均较好 ,其敏感度可达 3pg。应用该方法对 4例病变典型的小鹅瘟患鹅组织器官进行检测 ,结果表明 ,患鹅脏器中 ,小肠、心、肝、胰、脾组织含毒量较高 ,其中小肠组织含毒量最高 ,而脑组织中含毒量较低。应用琼脂扩散试验验证了核酸斑点杂交法的检测结果。     

Has the optimum time for faecal nematode egg count reduction testing in sheep in New Zealand changed?

Abstract

AIM: To determine what, if any, changes have taken place in the optimum time, for undertaking faecal egg count reduction tests (FECRT) in sheep in New Zealand.

METHODS: A comparison was made between the numbers and types of nematode genera adequately represented for testing purposes (faecal nematode egg count (FEC) of >50 epg) in initial FECRT case submissions to a veterinary laboratory in New Zealand, during two 4-year periods, in 1992–1995 and 2006–2009.

RESULTS: Although there were some minor differences between them, the seasonal patterns of occurrence remained basically the same for all parasite genera in both datasets, with their individual peak periods of representation occurring during February to May in all instances. Not surprisingly, this period of maximum seasonal occurrence for each parasite genus also coincided with those months of the year when the greatest numbers of worm genera were adequately represented for faecal nematode egg count reduction (FECR) testing.

CONCLUSIONS: The results of this study indicate that the optimum time for conducting FECRT in sheep in New Zealand continues to be during the late summer-autumn months of February to May. However, they also reaffirmed that even during this optimal period there are still likely to be many occasions when relatively few nematode genera may be sufficiently well represented for satisfactory FECR testing. Accordingly, veterinary practitioners ought to be aware that, in order to obtain a complete picture of the resistance status of all worm genera on a particular property, such testing may need to be carried out on more than one occasion.     

Associations among albuminuria, C-reactive protein concentrations, survival predictor index scores, and survival in 78 critically ill dogs

Background: Microalbuminuria and C‐reactive protein (CRP) are predictors of morbidity and survival in critically ill human patients. Hypothesis/Objectives: To evaluate results of microalbuminuria assays (untimed single‐sample urine albumin concentration [U‐ALB] and the urine albumin : creatinine ratio [UACR]), serum CRP, and survival predictor index (SPI2) scores as predictors of survival in critically ill dogs. Animals: Seventy‐eight dogs admitted to intensive care units at University of Tennessee (UT) and Colorado State University (CSU). Methods: Prospective observational study. Critically ill dogs were eligible for enrollment, unless euthanized because of financial constraints. Samples were collected within 3 hours of admission. Spearman's rank‐correlation coefficients were determined for U‐ALB, UACR, CRP, and SPI2. U‐ALB, UACR, CRP, and SPI2 were assessed for associations with 7‐ and 30‐day survival by Mann‐Whitney U‐tests and receiver operating characteristic (ROC) curves. P‐values < .0125 were considered significant. Results: UT (n = 49) and CSU (n = 29) patients did not differ significantly. Forty percent (31/78) of dogs died. SPI2 was inversely correlated with U‐ALB (r_s=?0.39, P < .001) and UACR (r_s=?0.41, P < .001). CRP was not correlated with SPI2 (P= .019), U‐ALB (P > .1), or UACR (P > .1). U‐ALB and UACR had very high correlation (r_s= 0.95, P < .001). SPI2, U‐ALB, and UACR differed significantly for survivors and nonsurvivors. SPI2, U‐ALB, and UACR had areas under the ROC curve (AUC) from 0.68 to 0.74 for survival prediction. Conclusions and Clinical Importance: Albuminuria and SPI2, but not CRP, are associated with survival in critically ill dogs. Suboptimal AUCs limit the value of microalbuminuria testing for clinical risk assessment. Additional studies are necessary to determine the usefulness of microalbuminuria testing in patient risk stratification for prospective research.     

一株鸡新城疫病毒的分离鉴定及其油乳剂灭活苗免疫效果评价

从河南某种鸡场发病鸡群中分离出1株毒株,经HA、HI和动物回归试验,确定分离的病毒为NDV,命名为HNYD株。经过测定,HNYD株鸡胚半数致死量（ELD50）为10^90/0．1mL;鸡胚最小致死量的平均致死时间（MDT）为53．8h;对1日龄SPF雏鸡脑内致病指数（ICPI）为1．86,提示HNYD株为强毒力株。血清交叉反应试验显示HNYD株和La Sota株存在一定的抗原性差异。用HNYD株油乳剂灭活苗对HNYD株和La Sota株的保护率分别为90％和100％,明显优于La Sota株油乳剂灭活苗,提示HNYD株制备的疫苗对流行毒株和经典毒株的攻击均能很好的保护。     

龙岩地区部分猪场弓形虫病的血清学调查

通过间接血凝(IHA)试验对采自龙岩地区12个猪场的2055份猪血清进行弓形虫病的抗体检测,结果表明猪弓形虫病在龙岩地区呈散发性流行。样品阳性率为5.99%,各猪场之间的抗体阳性率有一定差异,最高为16.22%,最低为0。在母猪群中随着胎次的增加,弓形虫病抗体阳性率也升高,由1～2胎的6.72%上升到5胎以上的12.18%;育肥猪群总体弓形虫病抗体阳性率较低,由哺乳仔猪的10.94%缓慢降至育肥猪的1.62%,呈现出随日龄增加弓形虫病抗体阳性率逐渐降低的趋势。     

地方、培育、引入品种资源的保存与发展的研究进展

作者就20世纪 80年代以来我国多次畜禽资源调查的结果作了详细的介绍,报道了576个品种在近30年来的变迁情况。除了常见的马、驴、黄牛、水牛、大额牛、牦牛、绵羊、山羊、猪、鸡、鸭、鹅、特禽的品种以外,尚有7种,如骆驼、兔、梅花鹿、马鹿、水貂、貉、蜂等动物。其中地方品种426个,培育品种73个,引入品种77个。已有19个地方品种灭绝,37个受严重威胁。作者回顾了20世纪末开始应用遗传检测手段,包括个体、细胞、蛋白质和分子水平4个层次的研究进展。证明多层次综合研究才能反映品种的全貌。建议资源调查中要着重选育强度带来的影响,品种调查提纲必须周密,要以品种特征和外貌为主,与遗传分析相结合,才可能提出可行的保种和开发方案。作者论证了配子和胚胎超低温保存、DNA保存的前景,并指出与企业合作进行保种的动向,国家应长期重点加以支持。     

An evaluation of the Abaxis VSPro for the measurement of equine plasma fibrinogen concentrations

Reasons for performing study: Accurate measurement of plasma fibrinogen concentrations is an important tool for assessment of horses with inflammatory diseases. Objectives: To determine the precision and accuracy of a benchtop instrument using both fresh and frozen equine plasma by comparing the plasma fibrinogen concentration measured by a benchtop instrument to 2 separate laboratory standard methods (ACL 100 and STA Compact) for fibrinogen measurement. Methods: Accuracy and precision of the VSPro was evaluated using both human fibrinogen standards and samples from horses. Fifty frozen samples from horses with gastrointestinal disease had the fibrinogen concentration measured using the ACL 1000 and the VSPro. Fifty fresh samples were collected from hospitalised horses and fibrinogen concentration was measured using the STA Compact coagulation machine and the VSPro. Correlations for measurements were performed, as well as Bland‐Altman analysis. Results: Coefficients of variability for the VSPro ranged from 7% to 15%. The VSPro fibrinogen values were well correlated to both the ACL 1000 (r = 0.94, P<0.001) and the STA Compact measurements (r = 0.926, P<0.001). Bland‐Altman analysis showed a mean bias of ‐0.83 g/l (95% confidence interval ‐2.03–0.324 g/l) for the ACL 1000 and a mean bias of ‐0.024 g/l (95% confidence interval ‐1.434–1.386 g/l) for the STA Compact. Conclusions: The VSPro appears to have adequate accuracy and precision for clinical measurement of plasma fibrinogen concentrations. Potential relevance: The VSPro provides a measurement of equine plasma fibrinogen concentration using a benchtop instrument with a rapid test time that has comparable accuracy to the fibrinogen concentration obtained from reference laboratories.     

入境肉种鸡检出衣原体阳性及其处理

目的随着加入世界贸易组织,我国海关检疫压力增大,为了维护国家主权和国际声誉,加强出入境检验检疫势在必行。衣原体病是一种重要的人畜共患病,也给养禽业带来巨大的经济损失,必须给予足够的重视,鹦鹉热衣原体对多种动物宿主有广泛的寄生性引起不同动物的多症候群疾病,是世界动物卫生组织动物名录疾病。方法对入境的一批肉种鸡进行抽样检验,按照中华人民共和国出入境检验检疫行业标准用间接血凝检测鹦鹉热衣原体病,做出试验报告。结果经检测出现阳性,肉鸡携带有衣原体病毒。结论根据中华人民共和国进出境动植物检疫法第二章进境检疫第十六条检出二类传染病、寄生虫病的动物,退回或者扑杀,同群其他动物在隔离场或者其他指定地点隔离观察。     

PRRSV重组N蛋白表达、纯化及间接ELISA检测试剂盒的研制

采用RT-PCR扩增出大小409bp猪繁殖与呼吸综合征病毒核衣壳蛋白基因（PRRSV N gene）,将该基因克隆到表达载体pGEX-KG,构建重组表达载体pGEX-KG-N,转化表达菌株BL21（DM3）诱导表达,经SDS-PAGE和Western blot分析表明,重组N蛋白获得了高效表达,融合蛋白相对分子质量为41 000,并具有良好的免疫学活性。扩大诱导培养,收集菌体,提取包涵体,用GST-Protein Purtification Kit亲和层析纯化目的蛋白,SDS-PAGE电泳检测纯度达到90%以上,可满足诊断抗原质量要求。用纯化PRRSV重组核衣壳蛋白GST-N为包被抗原,建立检测PRRSV血清的间接ELISA诊断方法,并组装ELISA试剂盒。优化后抗原最适包被质量浓度为2.5mg/L;血清最适稀释度为1∶100;对血清样本检测临界值为0.224;与美国IDEXX公司的HerdChek ELISA试剂盒符合率为92%,特异性为95%,敏感性为90%;与猪细小病毒、猪伪狂犬病病毒、猪乙型脑炎病毒、猪圆环病毒、猪瘟病毒等常见猪繁殖障碍病标准阳性血清无交叉反应。ELISA试剂盒批内和批间变异系数分别为3.44%～6.34%和5.04%～7.64%。置4℃条件保存12个月,试剂盒稳定性无明显改变。该试剂盒对350份临床疑似血清检出率为88.6%。研制的ELISA试剂盒为PRRSV临床血清抗体检测及流行病学调查提供了技术手段。