Paratuberculosis, a notifiable disease in Austria--current status, compulsory measures and first experiences

Paratuberculosis (Johne's disease) is one of the most important diseases in ruminants today. Its contribution is worldwide and the disease is causing severe financial losses among cattle producers in some countries [Hasanova, L., Pavlik, I., 2006. Economic impact of paratuberculosis in dairy cattle herds: a review. Vet. Med.-Czech. 51, 193–211]. Paratuberculosis is untreatable; diagnosis limited to the early stages of the infection and control of the disease is difficult. The prevalence of serologically positive Austrian cattle farms rose significantly to 19.0% during the past years [Baumgartner, W., Damoser, J., Khol, J.L., 2005. Comparison of two studies concerning the prevalence of bovine paratuberculosis (Johne's disease) in Austrian cattle in the years 1995–1997 and 2002/2003 (Article in German with extended English summary). Vet. Med. Austria/Wien. Tierärztl. Mschr. 92, 274–277]. Based on these findings clinical paratuberculosis in ruminants was declared a notifiable disease in Austria in April 2006.

A survey of the current situation in Austria, the most important parts of the new compulsory measures and their practical implementation and impacts are presented in this short communication.     

An Outbreak of Respiratory Disease Complex in Sheep in Central Ethiopia

An investigation was carried out into an outbreak of respiratory disease complex (RDC) in 3641 Menz and Awassi×Menz cross sheep in Central Ethiopia between 1998 and 1999 by clinical, serological, microbiological, post-mortem and histopathological examinations. The monthly incidence of RDC varied from 2.8% to 4.0% and the prevalence was as high as 17%. The case fatality rate was 18%, despite culling of sick sheep. Over 76% of the morbidity occurred in adults, followed by 19% among weaners. Similarly, 62% of the mortality was in adults. However, 27% of the mortality occurred in lambs despite the low morbidity in the group. Significant breed and age differences were seen in the morbidity and mortality rates (p<0.05). Clinical signs, gross and microscopic lesions and serological and bacteriological examinations showed an interplay of several causes of the RDC, including pestes des petits ruminants (PPR) (72.3%, serologically confirmed), lung worms, maedi-visna, bacterial bronchopneumonia (staphylococcal and streptococcal), enzootic pneumonia and some fungal infections. Cold temperatures, which may be as low as –8.5°C at night, are major predisposing factors along with managemental stresses. Vaccination of animals with a homologous PPR vaccine appeared to decrease dramatically the occurrence of the disease, showing that PPR played an important role in the outbreak. Several of the pathogens do not appear to be individually capable of causing the respiratory disease. Appropriate strategies for the prevention of RDC are suggested.     

Detection of Antibodies Against Peste des Petits Ruminants Virus in Sera of Cattle,Camels, Sheep and Goats in Sudan

Detection of antibodies against peste des petits ruminants virus in sera of cattle, camels, sheep and goats in Sudan.     

Use of Akaike information criteria for model selection and inference. An application to assess prevention of gastrointestinal parasitism and respiratory mortality of Guinean goats in Kolda,Senegal

A field experiment was carried out in Kolda (southern Senegal) from July 1986 to July 1988. Its goals were to: (1) describe the patterns of mortality of female Guinean goats by age, season and year; (2) assess preventive measures against respiratory diseases and gastrointestinal parasitism in reducing mortality; and (3) estimate the overall impact of these measures on survival to 1 year of age. Preventive measures for respiratory disease included vaccination against peste des petits ruminants (PPR) and pneumonic pasteurellosis (Pasteurella multocida types A and D). Control of gastrointestinal parasites was by deworming does with morantel (7.5 mg kg⁻¹, three times during the rainy season). The effects of vaccines and deworming were tested in a randomised factorial field experiment with villages being the experimental units. A total of 19 villages, 113 goat herds and 1458 goats were included in the study.

Generalised linear models of survival for five cohorts of goats (defined by five different birth seasons) used a binomial assumption for the response distribution and a complementary log–log link. Explanatory variables included age, season, year, vaccination, deworming and their interactions. A complex a priori model was built on the basis of previous epidemiological knowledge; a purposively selected set of simpler models was compared to this full model by the Akaike information criterion (AIC) and derived statistics. Inference on 1-year survival and treatment effects accounted for model-selection uncertainty. It was carried out with a bootstrap procedure and used information from the whole set of selected models.

Large variations in mortality by year and season were observed but no regular seasonal pattern was apparent. Mortality probabilities of kids in dewormed groups decreased quickly after birth, but remained elevated up to 9 months of age in the non-dewormed groups. Deworming lowered the risk of mortality. Vaccination alone was not protective (except during an observed outbreak of PPR).     

一步法实时定量RT-PCR检测小反刍兽疫病毒方法的建立

建立了检测小反刍兽疫病毒的快速、特异的基于Taqman的一步法实时定量RT-PCR方法。通过对GenBank已公布的小反刍兽疫病毒N基因序列进行序列比较分析,设计了一对特异性引物和一条Taqman探针。利用这对引物和探针对来自不同疫源地的小反刍兽疫病毒RNA样本进行检测,都获得了特异性扩增,但是,不与牛瘟病毒、海豚麻疹病毒等其他种的麻疹病毒属病毒发生交叉反应。本方法具有高度灵敏性,最低可检测到810拷贝的RNA模板。在模板浓度为8.1×102到8.1×108拷贝范围内,都呈现良好的线性关系,而且无论是组内和或组间重复的变异系数都很低。     

稳定表达山羊SLAM受体的BHK-21细胞系的建立及应用

信号淋巴激活分子(SLAM)又称为CD150,是小反刍兽疫病毒(PPRV)和犬瘟热病毒(CDV)等麻疹病毒属病毒感染淋巴细胞的主要受体,在病毒侵入细胞中发挥重要作用。为建立稳定表达山羊SLAM(g SLAM)的真核细胞系,本研究将人工合成的g SLAM基因克隆至真核表达质粒p IRESpuro3中,并在该基因的3'端引入Flag标签序列作为分子标记,构建了重组质粒p IRES3-g SLAM。将该重组质粒转染BHK-21细胞,经嘌呤霉素加压筛选及采用表达绿色荧光蛋白(GFP)的重组PPRV病毒(r PPRV/GFP)感染鉴定后,筛选到稳定表达g SLAM基因的细胞系。r PPRV/GFP感染和western blot鉴定表明,无论是否有嘌呤霉素压力的存在,该细胞系在传代至第20代,仍能稳定表达g SLAM蛋白。由于g SLAM氨基酸序列与犬的同源性较高,以表达GFP重组CDV强毒株(r CDV/GFP)感染该细胞系,病毒可以感染且能形成明显的细胞病变,表明该细胞系可用于CDV强毒分离和致弱机制等相关研究。     

Methods to determine resistance to anthelmintics when continuing larval development occurs

The in vivo faecal egg count reduction test (FECRT) is the most commonly used test to detect anthelmintic resistance (AR) in gastrointestinal nematodes (GIN) of ruminants in pasture based systems. However, there are several variations on the method, some more appropriate than others in specific circumstances. While in some cases labour and time can be saved by just collecting post-drench faecal worm egg counts (FEC) of treatment groups with controls, or pre- and post-drench FEC of a treatment group with no controls, there are circumstances when pre- and post-drench FEC of an untreated control group as well as from the treatment groups are necessary. Computer simulation techniques were used to determine the most appropriate of several methods for calculating AR when there is continuing larval development during the testing period, as often occurs when anthelmintic treatments against genera of GIN with high biotic potential or high re-infection rates, such as Haemonchus contortus of sheep and Cooperia punctata of cattle, are less than 100% efficacious. Three field FECRT experimental designs were investigated: (I) post-drench FEC of treatment and controls groups, (II) pre- and post-drench FEC of a treatment group only and (III) pre- and post-drench FEC of treatment and control groups.To investigate the performance of methods of indicating AR for each of these designs, simulated animal FEC were generated from negative binominal distributions with subsequent sampling from the binomial distributions to account for drench effect, with varying parameters for worm burden, larval development and drench resistance. Calculations of percent reductions and confidence limits were based on those of the Standing Committee for Agriculture (SCA) guidelines. For the two field methods with pre-drench FEC, confidence limits were also determined from cumulative inverse Beta distributions of FEC, for eggs per gram (epg) and the number of eggs counted at detection levels of 50 and 25. Two rules for determining AR: (1) %reduction (%R) < 95% and lower confidence limit <90%; and (2) upper confidence limit <95%, were also assessed. For each combination of worm burden, larval development and drench resistance parameters, 1000 simulations were run to determine the number of times the theoretical percent reduction fell within the estimated confidence limits and the number of times resistance would have been declared.When continuing larval development occurs during the testing period of the FECRT, the simulations showed AR should be calculated from pre- and post-drench worm egg counts of an untreated control group as well as from the treatment group. If the widely used resistance rule 1 is used to assess resistance, rule 2 should also be applied, especially when %R is in the range 90 to 95% and resistance is suspected.     

新疆小反刍兽疫疫情诊断

2013年12月新疆伊犁州霍城县发生不明山羊疫情,根据临床症状和剖检变化怀疑为小反刍兽疫感染。对3只病死山羊病料、8只患病山羊分泌物棉拭子样品和6只患病山羊血清样品分别进行病原学和血清学检测。利用竞争ELISA试剂盒对6份血清样本进行抗体检测,结果全部为阳性。利用抗原捕获ELISA试剂盒,在11只病羊样品中都检测到小反刍兽疫抗原。利用能特异性检测小反刍兽疫病毒的荧光定量RT-PCR方法,在11只病羊样品中检测到小反刍兽疫病毒核酸。利用特异引物进行PPRV N基因片段RT-PCR反应,从11只病羊样品中检测到PPRV核酸。针对2号样本病原核酸N基因和F基因片段进行序列同源性比较,结果该毒株与西藏流行株序列片段相似性分别为96.5%和97.5%。遗传进化分析,该病原属于谱系4,与巴基斯坦等国流行毒株遗传关系最近。     

苜蓿植株再生及PPRV-F基因转化条件的研究

以苜蓿品种甘农1号下胚轴为外植体,研究不同培养基对苜蓿愈伤组织、体细胞胚形成的影响,通过体细胞胚发生途径建立再生体系,在此基础上,摸索农杆菌介导小反刍兽疫病毒F基因的转化条件。结果表明:所选培养基愈伤组织诱导率均为100%,UM+0.1mg/L NAA+0.5mg/L KT为诱导体细胞胚的最佳培养基。农杆菌介导的苜蓿遗传转化条件为:下胚轴预培养3d,农杆菌菌液侵染15min,然后在培养基上铺一层灭菌滤纸共培养3d后清洗,选择压为卡那霉素(Km)75mg/L,抑菌浓度为头孢霉素(Cef)300mg/L。本研究为制备防治小反刍兽疫转基因植物疫苗奠定了基础。