Diagnosing feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV) infection: an update for clinicians

With the commercial release in Australia in 2004 of a vaccine against feline immunodeficiency virus (FIV; Fel‐O‐Vax FIV®), the landscape for FIV diagnostics shifted substantially. Point‐of‐care (PoC) antibody detection kits, which had been the mainstay for diagnosing FIV infection since the early 1990s, were no longer considered accurate to use in FIV‐vaccinated cats, because of the production of vaccine‐induced antibodies that were considered indistinguishable from those produced in natural FIV infections. Consequently, attention shifted to alternative diagnostic methods such as nucleic acid detection. However, over the past 5 years we have published a series of studies emphasising that FIV PoC test kits vary in their methodology, resulting in differing accuracy in FIV‐vaccinated cats. Importantly, we demonstrated that two commercially available FIV antibody test kits (Witness? and Anigen Rapid?) were able to accurately distinguish between FIV‐vaccinated and FIV‐infected cats, concluding that testing with either kit offers an alternative to PCR testing. This review summarises pertinent findings from our work published in a variety of peer‐reviewed research journals to inform veterinarians (particularly veterinarians in Australia, New Zealand and Japan, where the FIV vaccine is currently commercially available) about how the approach to the diagnosis of FIV infection has shifted. Included in this review is our work investigating the performance of three commercially available FIV PoC test kits in FIV‐vaccinated cats and our recommendations for the diagnosis of FIV infection; the effect of primary FIV vaccination (three FIV vaccines, 4 weeks apart) on PoC test kit performance; our recommendations regarding annual testing of FIV‐vaccinated cats to detect ‘vaccine breakthroughs’; and the potential off‐label use of saliva for the diagnosis of FIV infection using some FIV PoC test kits. We also investigated the accuracy of the same three brands of test kits for feline leukaemia virus (FeLV) diagnosis, using both blood and saliva as diagnostic specimens. Based on these results, we discuss our recommendations for confirmatory testing when veterinarians are presented with a positive FeLV PoC test kit result. Finally, we conclude with our results from the largest and most recent FIV and FeLV seroprevalence study conducted in Australia to date.     

肉用杂交犊牛小肠上皮细胞的分离培养和鉴定

本试验旨在建立肉用杂交犊牛小肠上皮细胞分离培养和鉴定的方法,为研究犊牛小肠上皮细胞营养吸收、免疫调控及肠道屏障功能提供原代细胞培养模型。选用新生未吮乳的肉用杂交犊牛的空肠组织,应用胶原酶Ⅰ和中性酶联合消化法对小肠上皮细胞进行分离,应用相差消化法和相差贴壁法对犊牛小肠上皮细胞进行纯化。通过细胞形态学观察、生长曲线、免疫荧光及染色体核型分析等方法来鉴定细胞。结果表明:1)应用胶原酶Ⅰ和中性酶联合消化法得到的犊牛小肠上皮细胞生长状况良好,经纯化后得到90%以上纯度的犊牛小肠上皮细胞,并稳定传代至10代以上; 2)犊牛小肠上皮细胞的生长曲线为"S"形,符合细胞增殖规律; 3)免疫荧光鉴定犊牛小肠上皮细胞特异性表达角蛋白13和绒毛蛋白; 4)透射电镜观察发现犊牛小肠上皮细胞边缘有微绒毛结构,细胞与细胞之间的紧密连接结构清晰可见; 5)染色体核型分析显示细胞内含有60条染色体,形态正常,呈二倍体核型。综上所述,应用胶原酶Ⅰ和中性酶联合消化及相差贴壁纯化成功得到犊牛小肠上皮细胞,为研究犊牛小肠上皮细胞营养物质吸收、免疫调控和肠道屏障功能提供了细胞素材。     

玉米秸秆青贮、羊草、燕麦草与精饲料组合效应的研究

本试验旨在利用体外产气法探究混合粗饲料(玉米秸秆青贮+羊草+燕麦草)与精饲料间的最优组合效应。试验采用单因素试验设计进行组合效应筛选,混合粗饲料玉米秸秆青贮∶羊草∶燕麦草为20∶30∶50,与精饲料以100∶0、80∶20、60∶40、50∶50、40∶60、20∶80以及0∶100的比例进行组合,每个组合3个重复。采用体外产气法分析累积产气量和不同组合比例时pH、干物质降解率(DMD)及微生物蛋白(MCP)、氨态氮(NH3-N)、挥发性脂肪酸(VFA)含量变化,计算各组合的单项组合效应指数和多项组合效应指数。结果表明:1)混合粗饲料和精饲料的不同组合比例对产气量均有显著影响(P<0.05),其中48 h产气量的单项组合效应指数在80∶20组最大。2)混合粗饲料和精饲料的不同组合比例对pH影响显著(P<0.05),且0∶100组最低。3)混合粗饲料和精饲料的不同组合比例对DMD影响显著(P<0.05),DMD的单项组合效应指数在20∶80组最大。4)NH3-N含量受混合粗饲料和精饲料的不同组合比例影响显著(P<0.05),以50∶50组NH3-N含量的单项组合效应指数最大。5)混合粗饲料和精饲料的不同组合比例对M CP含量具有显著影响(P<0.05),MCP含量的单项组合效应指数最高值出现在50∶50组。6)混合粗饲料和精饲料的不同组合比例对乙酸、丙酸、丁酸含量均有显著影响(P<0.05)。根据多项组合效应指数得出玉米秸秆青贮∶羊草∶燕麦草∶精饲料最优组合比例为10∶15∶25∶50。     

氮磷肥配施对西北羊茅开花期叶片光合特性日变化的影响

基于植物对N、P营养元素的需求,试验设计2个施氮量(0、60 kg·hm^-2)和3个施磷量(0、60、90 kg·hm^-2)配施,测定开花期叶片在不同光合有效辐射下的气体交换参数和叶绿素荧光参数,探讨西北羊茅对氮、磷添加和光合有效辐射的响应。结果表明,光合有效辐射呈先增后减的日动态变化,下午13:00时高达2000 μmol·m^-2·s^-1,且对西北羊茅光合特性的影响大于施肥措施。净光合速率随光合有效辐射增强先增后降,其值在800~1000 μmol·m^-2·s^-1时最大;气孔导度、气孔限制值、蒸腾速率和水分利用效率与净光合速率表现一致,胞间CO₂浓度表现相反。氮肥和磷肥配施后气孔导度、气孔限制值和蒸腾速率均增大,胞间CO₂浓度则减小,表明施肥主要通过调节气孔导度,以促进叶片与环境间交换水分和CO₂,从而增大净光合速率。随光合有效辐射增强,PSⅡ有效光化学效率、PSⅡ实际光化学效率和光化学猝灭系数显著降低,非光化学猝灭系数显著增加,电子传递速率先增加后降低,但均在400~800 μmol·m^-2·s^-1和大于1200 μmol·m^-2·s^-1后快速变化。氮肥和磷肥配施显著提高PSⅡ有效光化学效率、PSⅡ实际光化学效率、光化学猝灭系数和电子传递速率,N₆₀P₉₀处理还提高PSⅡ最大光化学效率和非光化学猝灭系数。在光能分配方面,为减轻强光对光合器官的破坏,随光合有效辐射增强光化学反应比例降低,天线热耗散和非光化学反应耗散比例增加。施肥主要通过非光化学反应比例减小补偿于光化学反应比例,从而提高光能利用率。综合以上结果,从西北羊茅叶片的光合特性来看,氮磷肥配施通过调节气孔导度,以促进叶片与环境间水分和CO₂的交换和提高PSⅡ反应中心活性,非光化学反应比例减小补偿于光化学反应比例,进而增加叶片净光合速率。     

基于PEDV ORF3基因的强弱毒株鉴别方法的建立及初步应用

为了建立一种快速鉴别PEDV强毒株和弱毒株的RT-PCR方法,针对PEDV ORF3基因设计一对引物,优化其反应体系和退火温度,并测试其特异性和敏感性。结果显示,在退火温度55.2℃,上、下游引物各0.5μL时PCR扩增效果最理想。特异性试验结果显示本方法对9种常见猪病病原的核酸模板均不产生扩增条带;敏感性试验显示该方法的检测下限为T-ORF3_IS 272拷贝/μL和T-ORF3_VC 14.4拷贝/μL。将本试验建立的RT-PCR法与已建立的双重RT-PCR方法进行比较,结果显示两种方法符合率为99.2%,且本法的实际检出率更高。对120份临床样本检测结果显示,PEDV强弱毒株的阳性率分别为87.18%和84.21%,其中9份样本中同时检出2种PEDV毒株,但并未对检测方法的准确性造成影响。建立了一种基于ORF3基因的RT-PCR检测法,该方法特异性强、灵敏度高,可准确区分PEDV野毒株和弱毒株。     

广东省首例非洲猪瘟疫情的诊断与疫点消毒效果评价

2018年12月17日,广东省珠海市某定点屠宰场发生该省首起非洲猪瘟疫情。诊断中发现,因病程不同,感染猪的临床症状和病理变化存在一定的差异,并非全部表现典型症状。疫情发生后,珠海市防控非洲猪瘟等动物疫病应急指挥部立即严格遵照《广东省非洲猪瘟突发疫情应急处置预案》的规定,采取了扑杀、消毒、发布封锁令,划定疫点、疫区和受威胁区等处置措施;封锁期内,按照《非洲猪瘟防治技术规范(试行)》规定,并参考联合国粮食及农业组织(FAO)技术标准,对疫点进行消毒,同时采集疫点内的环境样品,检测非洲猪瘟病毒(ASFV)核酸,评估消毒效果,为解除封锁提供依据。本文为全国非洲猪瘟疫情排查、临床诊断和处置提供了参考。     

整合素αvβ3在人工受精奶牛受孕前后表达规律研究

本试验通过对10头荷斯坦奶牛在受孕前后的整合素αvβ3表达变化规律与奶牛早期妊娠关系的研究,筛选早期妊娠检测标志性因子,从而提高奶牛的繁殖效率。试验表明,选取的10头荷斯坦奶牛中有7头受孕3头未受孕,检测发现,10、13、16、19d,αv基因和β3基因转录及整合素αvβ3表达水平在受孕奶牛血液中的表达量均高于未孕奶牛,特别是输精后第16d受孕奶牛的表达量显著高于未孕奶牛。因此,αvβ3检测可将早孕检测的时间窗口提前到人工输精后第16d。     

桔园主要害虫选择性药剂的系统筛选及其组合技术效益的研究

取40余种国内外农药,分别对柑桔害螨、蚜虫、蚧类和潜叶蛾等四大类主要害虫,采用多边测定、系统筛选的方法,选出尼索朗、螨死净、倍乐霸、托尔克、克螨特、卡死克、机油乳剂及叶蝉散等一批高效选择性或部分选择性药剂。大田试验结果表明,停用有机磷或有机磷和拟除虫菊酯类药剂,采用选择性药剂组合技术的实验区比常规区全年减少病虫防治费用14.91%-42.14%;同时,害虫天敌明显增加。据不完全统计,1988-1990年浙江桔区选择性药剂和部分选择性药剂的应用面积超过20余万公顷次;仅尼索朗一种药剂的使用面积就近11万公顷次,节约防治费用超过5000万元。研究结果阐明,在桔园内停止或控制使用对生态环境有严重破坏作用的有机磷和拟除虫菊酯类药剂是完全可行的,该项技术具有明显的经济、生态和社会效益。     

Diagnosis and treatment of biaxial keratomas in the left fore foot of a 15‐year‐old Clydesdale mare

A 15‐year‐old Clydesdale mare presented for further diagnostics and treatment of waxing and waning lameness and recurrent subsolar abscesses. Radiographs and computed tomography revealed biaxial masses extending from the hoof capsule, causing bone resorption of the distal phalanx. Surgery was performed to remove the masses and post operative care included regional limb perfusions, systemic antibiotics and therapeutic shoeing. Histopathology was consistent with the diagnosis of keratoma for each of the masses; this is the first case of confirmed biaxial keratomas. Two months after surgery the horse is sound at the walk and is expected to return to full function within the next year.