Equine gastric ulcer syndrome in adult donkeys: Investigation on prevalence,anatomical distribution,and severity

The study was performed on 39 live donkeys that underwent gastroscopic examination. The lesions were recorded in accordance with the European College of Equine Internal Medicine Consensus Statement guidelines. The presence of Gasterophilus sp. larvae was also recorded. Larvae were collected and identified to species level. Fisher's exact test was used to compare different prevalence values for sex, age, and anatomical distribution of lesions. Gastric lesions were present in 20/39 (51.3% [35.6–67%]) donkeys; 19/39 (48.7% [95% confidence interval = 33–64.4%]) were affected only by equine squamous gastric disease (ESGD), while 1/39 (2.6% [0–7.5%]) showed both ESGD and equine glandular gastric disease (EGGD), thus 95% of positive donkeys showed lesions located in the nonglandular mucosa. The ESGD grade was 0/4 (48.7% [33–64.4%]) in 19/39, 1/4 (12.8% [2.3–23.3%]) in 5/39, 2/4 (25.6% [11.9–39.5%]) in 10/39, 3/4 in 4/39 (10.3% [0.7–19.8%]) and 4/4 in 1/39 (2.6% [0–7.5%]) donkeys, respectively. The EGGD lesion was a mild depression in the ventral glandular fundus. ESGD was primary in all the donkeys included and lesions were located around the cardia and along the lesser curvature. Gasterophilus sp. larvae were present in all animals and were identified as third‐stage larvae of Gasterophilus intestinalis. No animals showed clinical signs of equine gastric ulcer syndrome (EGUS). No significant differences relating to sex, age or breed were found in the prevalence of EGUS in this study, while the proportion of donkeys affected by ESGD was statistically higher than those affected by EGGD. To the best of our knowledge, this is the first report on the gastroscopic evaluation of EGUS in live donkeys. Our results show a higher prevalence of EGUS in live donkeys than values previously reported by other authors in donkeys that were dead or had been subjected to euthanasia. The detection of third‐stage G. intestinalis larvae was not unexpected since these can be found in the stomach of equids throughout the year, and G. intestinalis has been reported as the most common Gasterophilus sp. in Italy.     

Phenylbutazone induces equine glandular gastric disease without decreasing prostaglandin E2 concentrations

In equids, phenylbutazone at high doses induces gastric disease, primarily in the glandular portion of the stomach. However, the mechanism of nonsteroidal anti‐inflammatory drug (NSAID)‐induced gastric disease in horses has yet to be determined. While phenylbutazone‐associated ulceration is often attributed to a decrease in basal gastric prostaglandins, this has not been demonstrated in the horse. Twelve horses were randomly assigned to treatment (n  = 6; 4.4 mg/kg phenylbutazone PO in 20 ml molasses q 12 hr for 7 days) or placebo (n  = 6; 20 ml molasses PO q 12 hr for 7 days) groups. Before treatment and 3 and 7 days after initiation of treatment, gastroscopy was performed and glandular gastric biopsies were collected and frozen at ?80°C. Glandular disease was assessed on a scale of 0–4. Prostaglandin E₂ concentrations in biopsies were measured using a commercially available enzyme‐linked immunosorbent assay. All phenylbutazone‐treated horses developed grade ≥2 glandular disease. Prostaglandin concentrations increased over time (p  = .0017), but there was no effect of treatment (p  = .49). These findings indicate that despite induction of glandular disease grade ≥2, phenylbutazone did not decrease basal glandular gastric prostaglandin E₂ concentration.     

含EGFP基因的马疱疹病毒1型新疆株gE基因缺失株的构建

为筛选马鼻肺炎（equine rhinopneumonitis,ER）基因缺失减毒活疫苗的候选毒株,本研究参考GenBank中EHV-1（登录号：KF644579.1）目的基因序列设计同源臂引物,以本地区流行株XJ2015株DNA为模板PCR扩增gE基因同源臂gEH1、gEH1,以EGFP表达盒（CMV+EGFP+polyA）为标记基因,酶切后依次连接至载体pUC-19,成功构建重组质粒pUC-gEH1H2-EGFP。将XJ2015基因组与质粒pUC-gEH1H2-EGFP共转染至RK-13细胞进行同源重组,以EGFP为标记进行gE基因缺失毒株的筛选及纯化,并测定重组毒株效价。结果显示：经5轮荧光噬斑纯化、PCR及测序鉴定,成功获取一株携带EGFP基因的重组毒株XJ2015-△gE-EGFP,且重组毒株效价（10^7.1TCID₅₀/0.1 mL）较原毒株（10^8.8TCID₅₀/0.1 mL）下降约10^1.7TCID₅₀/0.1 mL。采用同源重组技术成功构建了1株马疱疹病毒1型流行株gE基因缺失突变株,为未来筛选马鼻肺炎基因缺失弱毒疫苗奠定了基础。     

Comparative endoscopic evaluation of normal and ulcerated gastric mucosae in Thoroughbred foals

To contribute to early diagnosis and treatment of gastric ulcer of foals, we examined the gastric mucosa of healthy and affected foals using an endoscope. In healthy foals, the characteristic changes in the development of the squamous mucosa were seen mainly in the squamous mucosa, and maturation of the squamous mucosa in the greater curvature (GC-S) occurred more slowly than that of the squamous mucosa in the lesser curvature (LC-S). Epithelial desquamation in the LC-S and GC-S was observed between 6 and 90 days but was not observed in the LC-S at about 60 days, whereas it was observed in the GC-S until 90 days old. These findings suggest that there is a difference in the development of the gastric mucosa by region and that desquamation continues over a term longer than studies have reported in the past. In the affected foals, the minimum age at which gastric ulcer was observed was 4 days old. Gastric ulcers formed predominantly in the squamous mucosa (LC-S and GC-S) of foals with an immature mucosa before the weaning period, and the peak incidence occurred between 61 and 90 days old. The differences in the ulceration sites were considered to depend on the difference in the development (maturation) stage of the squamous mucosa. The grading score of the gastric ulcer increased with the growth of the affected foals. The gastric ulcer might be enhanced greatly by stress in the weaning period.     

MicroRNA fingerprints in serum and whole blood of sarcoid‐affected horses as potential non‐invasive diagnostic biomarkers

Serum and whole blood microRNA (miRNA) fingerprints have been proposed as a new class of non‐invasive human cancer biomarkers. In this study, we compared equine sarcoid (ES) disease‐specific serum and whole blood miRNA fingerprints and correlated them to miRNA expression in sarcoid tissue. After high throughput sequencing, miRNA differential expression analysis between six ES‐affected and five control horses was carried out in serum and whole blood using a DESeq algorithm, accounting for the influence of hemolysis and the white blood cell count. Target gene, pathway prediction and enrichment analyses were conducted using TarBase, mirPath and GeneCodis. After exclusion of 4 hemolyzed out of a total of 11 serum samples, 9 miRNAs were found to be differentially expressed in serum of ES vs control horses. In whole blood, all 11 samples showed normal white blood cell counts and 19 miRNAs were found to be differentially expressed. A total of 2/9 serum and 7/19 whole blood differentially expressed miRNAs were also highly expressed at the tissue level and their predicted target genes were associated with cancer pathways. Serum and whole blood miRNA expression allowed discrimination between ES and control horses and merits further validation in a larger study cohort. The use of whole blood might be superior because it has higher miRNA content and is less influenced by pre‐analytical variables compared to serum. Concurrent dysregulation of single miRNAs in tissue and blood suggests a possible biological function of circulating miRNAs.     

Combined decongestive therapy including equine manual lymph drainage to assist management of chronic progressive lymphoedema in draught horses

Equine chronic progressive lymphoedema (CPL) is a disabling disorder of draught horse breeds. Combined decongestive therapy (CDT) is the treatment of choice for lymphoedema in man and has been adapted for use in horses. Equine CDT, which includes manual lymph drainage (MLD) and subsequent bandaging with short stretch bandages, was expected to improve the signs of CPL in draught horses because CPL resembles primary lymphoedema in man. Five affected horses ‐ Gypsy cob (n = 1), Clydesdale (n = 1), Shires (n = 3) ‐ were included. Lesions were documented pre‐ and post treatment. Percentage volume loss of the distal legs was calculated using the disc model. Initial plans for daily CDT had to be adapted; intermittent treatment of Chorioptes infections required alternating between CDT and MLD in 4/5 horses. Concurrent pyoderma (1/5 horses) was treated throughout the study. Development of unrelated lameness (hoof abscess) allowed limited CDT treatment only in one horse. Marked softening of previously firm tissue indicated the change from ‘brawny’ to pitting oedema in 2/5 horses. Fibrotic nodules and folds in the pasterns became markedly softened and smaller in 2/5 horses. Skin surface notably improved in all horses: hyperkeratosis decreased, erosions and ulcerations healed completely and crusts disappeared. After 2 weeks, a mean volume reduction of 11.25% was seen, ranging from 4.75–21.74% and quality of movement improved. This pilot study documents evidence that CDT assists management of CPL. Current CPL management is limited to palliative treatments of secondary infections. Whilst not a permanent treatment, CDT offers a promising tool to manage horses with CPL, improving their quality of life and potential usefulness. More extensive and prolonged studies with a larger number of horses are warranted to evaluate the full potential of CDT.     

Time-related Pathological Changes in Horses Experimentally Inoculated with Equine Influenza A Virus

To investigate the pathology of equine influenza, necropsy of 7 horses experimentally infected with equine influenza A virus (EIV) subtype H3N8 was conducted on post-infection days (PID) 2, 3, 7, and 14. Histopathologically, rhinitis or tracheitis including epithelial degeneration or necrosis with loss of ciliated epithelia and a reduction in goblet cell numbers, was observed in the respiratory tracts on PIDs 2 and 3. Epithelial hyperplasia or squamous metaplasia and suppurative bronchopneumonia with proliferation of type II pneumocytes were observed on PIDs 7 and 14. Viral antigen was detected immunohistochemically in the epithelia of the nasal mucosa, trachea, and bronchi on PIDs 2 and 3. The sodA gene of Streptococcus equi subsp. zooepidemicus, a suspected cause of suppurative bronchopneumonia, was detected in paraffin-embedded lung tissue sections, but only on PIDs 7 and 14. These findings suggest that damage caused to ciliated epithelia and goblet cells by EIV infection results in secondary bacterial bronchopneumonia due to a reduction in mucociliary clearance.     

Viruses associated with outbreaks of equine respiratory disease in New Zealand

AIM: To identify viruses associated with respiratory disease in young horses in New Zealand.

METHODS: Nasal swabs and blood samples were collected from 45 foals or horses from five separate outbreaks of respiratory disease that occurred in New Zealand in 1996, and from 37 yearlings at the time of the annual yearling sales in January that same year. Virus isolation from nasal swabs and peripheral blood leukocytes (PBL) was undertaken and serum samples were tested for antibodies against equine herpesviruses (EHV-1, EHV-2, EHV-4 and EHV-5), equine rhinitis-A virus (ERAV), equine rhinitis-B virus (ERBV), equine adenovirus 1 (EAdV-1), equine arteritis virus (EAV), reovirus 3 and parainfluenza virus type 3 (PIV3).

RESULTS: Viruses were isolated from 24/94 (26%) nasal swab samples and from 77/80 (96%) PBL samples collected from both healthy horses and horses showing clinical signs of respiratory disease. All isolates were identified as EHV-2, EHV-4, EHV-5 or untyped EHV. Of the horses and foals tested, 59/82 (72%) were positive for EHV-1 and/or EHV-4 serum neutralising (SN) antibody on at least one sampling occasion, 52/82 (63%) for EHV-1-specific antibody tested by enzyme-linked immunosorbent assay (ELISA), 10/80 (13%) for ERAV SN antibody, 60/80 (75%) for ERBV SN antibody, and 42/80 (53%) for haemagglutination inhibition (HI) antibody to EAdV-1. None of the 64 serum samples tested were positive for antibodies to EAV, reovirus 3 or PIV3. Evidence of infection with all viruses tested was detected in both healthy horses and in horses showing clinical signs of respiratory disease. Recent EHV-2 infection was associated with the development of signs of respiratory disease among yearlings [relative risk (RR)=2.67, 95% CI=1.59-4.47, p=0.017].

CONCLUSIONS: Of the equine respiratory viruses detected in horses in New Zealand during this study, EHV-2 was most likely to be associated with respiratory disease. However, factors other than viral infection are probably important in the development of clinical signs of disease.     

TRANSCUTANEOUS ULTRASONOGRAPHIC EVALUATION OF THE AIR-FILLED EQUINE STOMACH AND DUODENUM FOLLOWING GASTROSCOPY

Gastroscopy with air insufflation was performed in 10 ponies, after which a transcutaneous ultrasound examination of the stomach and duodenum was performed immediately and at 1, 2, and 4 h postgastroscopy, and 24 h after feeding. Stomach measurements included the dorsoventral and craniocaudal dimensions, as well as the stomach depth from the skin surface and stomach wall thickness at the different time periods. Gastric wall folding was observed in one pony, becoming most distinct 2–4 h postgastroscopy. An undulating stomach wall was noted in eight other ponies postgastroscopy. These observations appeared to be a response to the deflation of the stomach as the insufflated air was released gradually. Gas was detected in the duodenum after the gastroscopy. The parameters measured were noted to be useful to evaluate the extent of stomach distension due to air or feed. The ultrasonographic appearance of the stomach can, therefore, be altered by gastroscopy and this should be borne in mind when examining horses with suspected gastric disease.     

马疱疹病毒1型LAMP检测方法的建立

本试验旨在建立一种检测马疱疹病毒1型（EHV-1）的快速、灵敏、特异的环介导等温扩增技术（LAMP）,同时评价该方法的可靠性。根据马鼻肺炎糖蛋白B（gB）基因特异保守序列设计多对LAMP引物,利用LAMP Real Time Turbidimeter LA-320仪监测反应进程,进行引物筛选和反应条件的优化,建立能特异性扩增EHV-1 DNA的LAMP检测方法,并加入SYBR GreenⅠ通过肉眼判断结果。该方法在65 ℃恒温下作用50 min,使EHV-1 DNA获得了高效率的特异性扩增,与其他马易感病毒如马疱疹病毒4型（EHV-4）等无交叉反应;且具有极高的灵敏性,可检测到10^-4稀释的目标病毒,比普通PCR的灵敏度高10倍;反应结束后加入SYBR GreenⅠ肉眼观察的结果与LAMP Real Time Turbidimeter LA-320仪监测结果一致。通过将4份临床样品的LAMP检测结果与已得到验证的PCR结果进行比对,结果显示符合率为100%。本研究建立的LAMP检测方法具有快速、特异、灵敏、简单易操作且设备要求低等特点,具有实地检测EHV-1的前景。     

Pathology of gastritis and gastric ulceration in the horse. Part 1: Range of lesions present in 21 mature individuals

Reasons for performing study: Gastric ulceration is now widely recognised as an important disease in high performance horses. Little is known about gastric histopathology in healthy or diseased animals; a comprehensive assessment would enhance interpretation of gross findings through assessment of their accuracy and allow for identification of lesion variety and pathogenesis in different anatomical regions of the stomach. Objectives: To investigate the true extent and variety of gastric lesions in a mixed population of mature horses at post mortem. Methods: Stomachs were removed from a mixed population of 21 horses at post mortem. Mucosal abnormalities were recorded in photographic and written form. Representative samples from all gross lesions were taken for histopathology and processed routinely. Special stains including Gram, PAS and Warthin Starry, were used when appropriate. Pathological classification of lesion type using both gross and histological appearances was performed. Results: Classification of lesions within the squamous region included hyperkeratosis, punctate scars, diffuse erosions/ulcerations and margo injuria; and within the glandular region, hyperaemia, focal erosions and ulcerations. Glandular metaplasia was recognised for the first time in the equine stomach. No Helicobacter‐like organisms were detected in association with lesion development. Conclusions and potential relevance: This study used gross and histological examination to highlight the large variety of naturally occurring gastric lesions in a mixed population of horses. Analysis of the pathogenesis of lesion development is now possible. Further research regarding the range of pathology in larger, more diverse groups of horses is required.     

Epidemiology of Mycobacterium bovis infection in feral ferrets (Mustela furo) in New Zealand: I. Pathology and diagnosis

Necropsies of 228 ferrets captured from eight areas in the North and South Islands provided material for an investigation into the epidemiology of tuberculosis in feral ferrets. Mycobacterial culture of pooled lymph nodes (retropharyngeal, respiratory and jejunal) identified the prevalence of infection to be much higher than that estimated from gross lesions only. Seventy-three of the 228 animals examined (32%) were diagnosed as tuberculous. Fifty-three culture-positive ferrets and 18 seemingly uninfected animals were subjected to detailed histopathological examination. The outcomes of these investigations, including the characteristics of the disease, distribution of lesions and aids to diagnosis, are presented.

Of the feral carnivores found in New Zealand, the disease persists at high prevalence only in ferrets, and is probably maintained principally by ingestion of tuberculous carrion. The course of the disease may be prolonged in some ferrets, but tuberculosis eventually causes the death of many infected animals. Microscopic hepatic granulomas may be considered pathognomonic of the disease, and have potential to be used as a rapid diagnostic tool in ferrets with no gross lesions.     

马传染性贫血病毒驴白细胞弱毒疫苗株囊膜基因的优化及其DNA疫苗体外瞬时表达研究

马传染性贫血病毒(Equine infectious anemia virus,EIAV)的密码子使用频率与哺乳动物间存在着明显差异。为此,对马传染性贫血病毒驴白细胞弱毒疫苗株(DLA-EIAV)囊膜全长基因按照哺乳动物优势密码子的使用原则进行了重新设计和合成,并以此为基础通过重叠延伸PCR、限制酶切等方法得到结合型和分泌型囊膜基因,将其插入含有鸡beta-actin/兔beta-globin复合启动子(AG)的高效表达载体pCAGGS中,构建了EIAV驴白细胞弱毒疫苗株结合型和分泌型囊膜基因的DNA疫苗质粒pCAGGS-opti-bou-env、pCAGGS- opti-sec-env。将构建的质粒纯化后分别转染293T细胞,以间接免疫荧光和Western blot方法检测转染48 h后细胞及上清中囊膜蛋白的表达。结果显示,两种表达质粒均可正确表达EIAV囊膜蛋白,与相对应未优化的表达载体pCAGGS-wt-bou-env和pCAGGS-wt-sec—env相比,密码子优化的基因体外瞬时表达水平有极为显著的提高,而且蛋白表达部位也与预期的结果符合。这一结果为EIAV囊膜蛋白的单抗制备、表位鉴定、免疫试验、新疫苗的开发等奠定了基础。     

山东省部分养殖场点马属动物主要病毒疫病调查

为了解山东省马属动物主要病毒疫病流行现状,采集青岛市2个马场以及济南、聊城和德州等驴养殖密集地市4个驴场的血清和全血样品各694份,分别进行马传染性贫血、H3N8亚型马流感血清抗体检测和非洲马瘟病原检测。结果显示:马传染性贫血、非洲马瘟检测全部为阴性,H3N8亚型马流感群体阳性率和样品阳性率分别为100%(6/6)和81.38%(520/639)。结果表明:马传染性贫血和非洲马瘟在这些养殖场点已得到很好的预防,但鉴于国外严峻的疫情形势,需要继续加强监测,防止病原传入;H3N8亚型马流感流行较为普遍,感染率较高,需要重点加强防控。本调查为山东省马属动物疫病防控提供了数据参考。