Pack wounds of donkeys and mules in the Northern High Atlas and lowlands of Morocco

Reasons for performing study: Wounds are one of the primary welfare concerns of working equids and are often related to harness or load‐bearing packs. Objectives: To identify and quantify factors associated with the presence of pack wounds in donkeys and mules working in the wood trade in the Midelt (Northern High Atlas) region of Morocco, and to compare wounds of these donkeys and mules to those working in other trades. Methods: The study population comprised equids working in the Midelt region involved in the wood trade. Comparison populations were comprised of equids working in other trades, in the same region and in Khemmiset region (lowlands of Central Morocco). Wounds were numbered and scored according to their severity. Questionnaires were used to gather data from owners, and objective data regarding the animal, berdaa (back‐pack) and load were collected. Data were analysed using multilevel uniand multivariable regression models to allow for clustering of wound sites within an equid. Results: Analysis showed an overall pack wound prevalence of 54% (n = 80/147). The most common site was the withers. Equids working in the wood trade were at increased risk of suffering pack wounds. Equids in the lowland study population were at increased risk of developing pack wounds, and donkeys were at increased risk of suffering pack wounds compared to mules both overall and within the wood trade. Other factors affecting the prevalence of wounds included the cleanliness of the berdaa and the frequency and distances of work imposed on the equid. Conclusion: Pack wounds in donkeys and mules are common in the 2 study regions of Morocco and appear to be associated with work factors including the trade the equids work in, the frequency and distances of work imposed and cleanliness of the pack. Donkeys are more susceptible than mules to developing pack wounds. Potential relevance: These results allow recommendations to be made to owners of equids that may decrease the prevalence of wounds and, therefore, improve welfare and working capability of these equids. Research is required to identify reasons for regional difference in prevalence of pack wounds in equids not working in the wood trade.     

Equine infectious anemia in mules: virus isolation and pathogenicity studies

There appears to be a lack of information concerning responses of mules to natural infection or experimental inoculation with equine infectious anemia virus (EIAV). In the present study EIAV was isolated from mules, for the first time, and its pathogenicity in naturally infected and experimentally inoculated animals was investigated. Two naturally infected (A and B) and three EIAV free mules (C, D and E) were used for this purpose. Mule A developed clinical signs, whereas mule B remained asymptomatic until the end of the study. Mules C and D were each inoculated with 10ml of blood from mule A and developed signs of the disease; they were euthanatized or died at day 22 and 25 post-inoculation, respectively. Mule E served as a negative control. The virus was isolated from the plasma samples of mules with clinical signs of the disease (A, C and D), but not from the asymptomatic mule B. Both proviral DNA and viral RNA were amplified from blood and tissues of the infected animals by nested polymerase chain reaction (nPCR). Antibodies were not detected in the two experimentally infected mules until their natural death or euthanasia. Clinicopathological and laboratory findings showed that, in mules, EIAV produced clinical signs similar to those observed in horses and ponies. Nested PCR proved to be a rapid, sensitive and specific diagnostic method for the detection of EIAV, regardless of the disease stage.     

Common helminth infections of donkeys and their control in temperate regions

Roundworms and flatworms that affect donkeys can cause disease. All common helminth parasites that affect horses also infect donkeys, so animals that co‐graze can act as a source of infection for either species. Of the gastrointestinal nematodes, those belonging to the cyathostomin (small strongyle) group are the most problematic in UK donkeys. Most grazing animals are exposed to these parasites and some animals will be infected all of their lives. Control is threatened by anthelmintic resistance: resistance to all 3 available anthelmintic classes has now been recorded in UK donkeys. The lungworm, Dictyocaulus arnfieldi, is also problematical, particularly when donkeys co‐graze with horses. Mature horses are not permissive hosts to the full life cycle of this parasite, but develop clinical signs on infection. In contrast, donkeys are permissive hosts without displaying overt clinical signs and act as a source of infection to co‐grazing horses. Donkeys are also susceptible to the fluke, Fasciola hepatica. This flatworm can be transmitted, via snails and the environment, from ruminants. As with cyathostomins, anthelmintic resistance is increasing in fluke populations in the UK. A number of the anthelmintic products available for horses do not have a licence for use in donkeys, and this complicates the design of parasite control programmes. As no new equine anthelmintic classes appear to be near market, it is important that the efficacy of currently effective drugs is maintained. It is important that strategies are used that attempt to preserve anthelmintic efficacy. These strategies should be based on the concept that the proportion of worms in a population not exposed to anthelmintic at each treatment act as a source of ‘refugia’. The latter is an important factor in the rate at which resistance develops. Thus, it is imperative that parasite control programmes take into account the need to balance therapy to control helminth‐associated disease with the requirement to preserve anthelmintic effectiveness.     

中国马传染性贫血病毒DLV株前病毒gp90基因在马体内的变异分析

分别以EIAV辽宁马强毒株（EIAVliao）前病毒DNA和马传染性贫血驴白细胞弱毒疫苗株（EIAVDLV）前病毒DNA为模板,从免疫接种后不同时期马血清中利用nested—PCR技术,扩增了约1．4kb的gp90基因。将其克隆后进行了测序,测序结果表明,免疫EIAVDLV后的第1时期至第5时期,与EIAV—liao比较核苷酸序列平均差异率分别为3．2956％、3．1456oA、3．36％、3．1856％和3．6456％。EIAVliao有20个N-连接糖基化位点,EIAVDLV平均是17．2个,其中有11个N-连接糖基化位点是高度保守的。     

马传贫弱毒疫苗接种马(骡)免疫反应动力学研究

本试验对注射马传贫弱毒疫苗后的马(骡)50匹,分别进行了体温、血液学、血清抗体及血液中T淋巴细胞等项目的检测。在此基础上,探讨了注苗后马(骡)免疫反应及其动态规律。提出注苗马(骡)在T细胞恢复到正常水平前,仍有感染强毒的可能,因此对马(骡)仍需进行监控。     

马传贫驴白细胞弱毒疫苗株跨膜蛋白主要免疫决定区基因的克隆与表达

从感染驴白细胞的马传贫驴白细胞弱毒疫苗株前病毒DNA中克隆了编码跨膜蛋白主要免疫决定区(TMIR)的基因，并在大肠杆菌中进行了表达。所表达的融合蛋白有一部分是可溶的，其氨基端带有6个组氨酸的标签，因此可以用固定化金属离子亲和层析法在非变性条件下进行纯化。在间接酶联免疫吸附试验(ELISA)和免疫印迹试验中，重组的TMIR蛋白可与马传贫阳性血清样品发生反应，而与健康马血清无任何反应。这表明该重组蛋白具有良好的抗原性和特异性，可用于马传贫弱毒疫苗株在体内外复制、接种马体内免疫应答及马传贫诊断的研究。     

马传染性贫血病毒第19、26代驴胎皮肤细胞弱毒前病毒DNA全基因序列分析

不同代次马传染性贫血驴胎皮肤细胞弱毒(Fetal donkey dermal virus,FDDV)的免疫保护效果各不相同,只有第10～15代驴胎皮肤细胞弱毒具有良好的免疫保护效果,可作为疫苗使用,继续传代则疫苗的保护率下降。为确定有、无免疫保护效果的FDDV在基因水平上的差异,本实验对无免疫保护效果的第19、26代驴胎皮肤细胞弱毒前病毒DNA进行了全基因序列测定,并与已测序的疫苗毒株进行序列比较。第19代和第26代FDDV全基因核苷酸序列同源性高达99.5%,与疫苗毒株全基因核苷酸序列的同源性分别为96.9%、96.7%。LTR是EIAV在细胞传代中变异最显著的区域,第19、26代FDDV的LTR与疫苗毒的LTR同源性仅为89.6%、89.3%。马传贫病毒的gag基因高度保守,第19、26代FDDV与疫苗毒株的gag基因推导氨基酸序列仅有2个氨基酸不同。第19、26代FDDV与疫苗毒株的pol基因、env基因的推导氨基酸序列的同源性分别为98.9%、98.8%、93.7%、93.6%。由序列比较结果可以推断,第19代、第26代FDDV不具有免疫保护效果的主要原因可能是由于LTR和env基因的变异,导致病毒复制能力下降或免疫原性丧失,不能诱导机体产生良好的免疫反应。     

Serodiagnosis of equine infectious anemia by agar gel immunodiffusion and ELISA using a recombinant p26 viral protein expressed in Escherichia coli as antigen

We used a p26 recombinant protein (p26r) from equine infectious-anemia virus (EIAV) expressed in Escherichia coli as antigen to standardize an agar-gel immunodiffusion (AGIDp26r) test and an indirect ELISA (ELISAp26r) for the detection of antibodies against EIAV in 720 equine sera from Brazil. We evaluated the tests's relative diagnostic sensitivities (relSe) and relative diagnostic specificities (relSp) against a commercial AGID kit (Idexx™, USA). We used three sera panels: panel A—196 AGID-negative sera from an AIE non-endemic controlled area; panel B—194 AGID-negative sera from an AIE endemic area and panel C—330 AGID-positive sera from an AIE endemic area. ELISAp26r cut-off value was defined with TG-ROC using sera from panels A and C. AGIDp26r showed an agreement of 100% with the commercial kit. When applied to sera from panels A and C, ELISAp26r showed an agreement of 100% with the kit, but, although relSe was 100% for panel C, the ELISAp26r had relSp of 93.3%.     

爱滋病毒抗原(P_(24))检测马传染性贫血病毒的研究

本研究通过基因工程技术生产了爱滋病毒的Ｐ２４抗原,并且运用于马传贫病毒的血清学检查,与传统方法所制备的马传贫抗原对照检测军马２５２４匹以及本实验室保留的各种马传贫阳性血清２２４份。结果显示马传贫血清能够免疫沉淀爱滋病毒（ＨＩＶ）的Ｐ２４,表明它们之间有着共同的抗原决定簇,且Ｐ２４检测法检测率高,检测周期仅３～４ｈ,明显优于传统法,可用于马传贫的快迅检测。     

A comparative study of xylazine-induced mechanical hypoalgesia in donkeys and horses

ObjectiveTo compare the effects of xylazine on mechanical nociceptive thresholds in donkeys and horses.Study designRandomized, controlled, crossover, Latin-square, operator-blinded design.AnimalsSix 3.1 ± 0.89 year old standard donkeys weighing 145.0 ± 30.5 kg and six 9.6 ± 4.4 year old Thoroughbred horses weighing 456.0 ± 69.0 kg.MethodsEach animal received one of four doses of xylazine (0.5, 0.7, 0.9, and 1.1 mg kg^?1), or acepromazine (0.05 mg kg^?1) or saline solution (0.9%) intravenously and mechanical nociceptive thresholds were assessed over 90 minutes. The areas under the threshold change versus time curve values for 60 minutes (AUC_0-60) post-drug administration were used to compare the effect of treatment. A 1-week interval was allowed between successive trials on each animal.ResultsAll doses of xylazine, but not acepromazine or saline, increased mechanical thresholds for up to 60 minutes. Xylazine-induced hypoalgesia was dose-dependent and corresponding AUC_0-60 values for each treatment were not significantly different between donkeys and horses (p≥ 0.0697).ConclusionThe hypoalgesic effects of xylazine at four different doses were not different between donkeys and horses.Clinical relevanceXylazine induced a similar degree of mechanical hypoalgesia in donkeys and horses suggesting that similar doses are needed for both species with regard to analgesia.     

马传染性贫血病毒疫苗株S2基因不同变异位点逆向突变感染性克隆的体外复制特性分析

为研究EIAV弱毒疫苗株(EIAV_(FCCV15))S2基因发生的稳定性突变对疫苗株特性的作用以及S2基因的功能,以EIAVFDDVl5感染性克隆质粒pFDDV3-8为模板,根据疫苗研制过程中S2基因发生的4个主要稳定性变异位点,构建及拯救出S2基因不同差异位点逆向突变为强毒株相应氨基酸的6株感染性克隆衍生毒株.经实时定量PCR、逆转录酶活性和western blot等检测表明,疫苗株S2基因4个稳定性变异位点全部逆向突变的感染性克隆衍生毒、,pFDDVS2rl-3-4-5在体外培养靶细胞中的复制水平低于亲本疫苗株感染性克隆衍生病毒和其他组合的逆向突变的感染性克隆衍生病毒,提示ELAV疫苗株S2蛋白4个稳定突变位点的综合作用可能是决定疫苗株和强毒株特性差异的因素之一.以上结果为体内感染S2基因逆向突变感染性克隆衍生病毒,进一步揭示S2基因在ELAV弱毒疫苗致弱过程中的作用提供了依据.     

Seroprevalence of Fasciolosis in Equines of the Black Sea Region,Turkey

This study determined the seroprevalence of fasciolosis in equines (n = 140) by enzyme-linked immunosorbent assay and Western blot. Blood samples were collected from different locations of the Black Sea Region of Turkey. The overall seroprevalence of fasciolosis in equines was 19.3%: 18% of horses (15), 22.6% of donkeys (7), and 19.2% of mules (5). The seroprevalence rates across provinces were as follows: 25% (5/20) in Samsun, 21.9% (7/32) in Tokat, 31.4% (11/35) in Amasya, 7.4% (2/27) in Ordu, and 7.7% (2/26) in Sinop. Three protein bands with molecular weights of 13, 30-40, and 40-65 kDa were determined in antigen clusters in sera from Fasciola spp-infected equine animals.     

Molecular detection of equine infectious anemia virus in clinically normal, seronegative horses in an endemic area of Mexico

Equine infectious anemia (EIA) is a highly infectious disease in members of the Equidae family, caused by equine infectious anemia virus (EIAV). The disease severity ranges from subclinical to acute or chronic, and causes significant economic losses in the equine industry worldwide. Serologic tests for detection of EIAV infection have some concerns given the prolonged seroconversion time. Therefore, molecular methods are needed to improve surveillance programs for this disease. We attempted detection of EIAV in 6 clinical and 42 non-clinical horses in Nuevo Leon State, Mexico, using the agar gel immunodiffusion (AGID) test for antibody detection, and nested and hemi-nested PCR for detection of proviral DNA. We found that 6 of 6, 5 of 6, and 6 of 6 clinical horses were positive by AGID, nested PCR, and hemi-nested PCR, respectively, whereas 0 of 42, 1 of 42, and 9 of 42 non-clinical horses were positive by these tests, respectively. BLAST analysis of the 203-bp 5′-LTR/tat segment of PCR product revealed 83–93% identity with EIAV isolates in GenBank and reference strains from other countries. By phylogenetic analysis, our Mexican samples were grouped in a different clade than other sequences reported worldwide, indicating that the LRT/tat region represents an important target for the detection of non-clinical horses.     

马传染性贫血病毒弱毒疫苗S2基因在体内变异分析

为研究马传染性贫血病毒(EIAV)驴白细胞弱毒疫苗EIAVDLV121的S2基因在马体内的变异规律,本研究选用4匹成年马,其中2匹(#1、#2)接种EIAVDLV121,另外2匹作为对照.免疫后监测马体温、血小板含量以及病毒载量结果显示,免疫马未出现马传染性贫血体征.通过RT-PCR方法检测病毒S2基因在感染马体内不同时期的基因序列,结果显示,免疫马体内EIAVDLV121 S2蛋白的突变主要发生在氨基酸第17位、22位、39位、41位、51位和55位.另外,#1马免疫后70 d以及#2马免疫后第14 d和第28 d检测疫苗毒S2蛋白序列与EIAVDLV121亲缘关系较近,而#1马免疫后第42 d、第112 d和第140 d的疫苗毒S2蛋白序列与EIAVDLV121的致病性亲本强毒株EIAVDN40亲缘关系最近.本研究结果有助于对EIAV及EIAV疫苗株在马体内感染进程的研究.     

马传染性贫血病毒自然感染马LTR序列分析

我国从20世纪70年代开始使用EIAV弱毒疫苗后,在全国范围内基本控制了马传贫的发生,我们从现地少数EIAV琼扩阳性马外周血中扩增并克隆了二株EIAV前病毒5'LTR序列,并与国内外毒株5'LTR序列进行了比较分析,发现中国EIAV LTR特有的特异性细胞转录因子结合基序,同时发现PEA2位点不是强毒特有的基序,在弱毒中亦发现该基序.     

Investigating the opinions of donkey owners and veterinary surgeons towards pain and analgesia in donkeys

The aim of this questionnaire‐based study was to identify which analgesics are currently prescribed to donkeys in clinical practice and to collect opinions from veterinary surgeons and donkey owners on the importance of different pain‐related behaviours in donkeys. Questionnaires were completed by 143 veterinary surgeons and 93 donkey owners. Phenylbutazone, flunixin, detomidine and butorphanol were reported as the most frequently used analgesics in donkeys. Most veterinary surgeons reported administering these drugs at the horse dose rate and dosing interval, which may be inappropriate in the donkey. Four pain‐related behaviours were identified as very important by veterinary surgeons and owners: dullness for colic‐associated pain, keeping the foot lifted, and lameness for foot and limb pain and inability to chew properly for head and dental pain. This is a more limited behavioural repertoire than has recently been identified to be associated with pain in the donkey and suggests that the respondents were reporting perceived wisdom as to what people think are the behavioural signs associated with pain or were not interpreting pain‐related behaviours correctly.