Polymerase Chain Reaction (PCR) is a molecular biology technique used to amplify specific DNA sequences, allowing for detailed genetic analysis in horses. This method enables the detection and quantification of genetic material, facilitating research in areas such as genetic disorders, infectious diseases, and population genetics in equine species. PCR applications in horses include identifying pathogens, verifying parentage, and studying genetic variations. The technique's sensitivity and specificity make it a valuable tool in equine veterinary diagnostics and research. This page compiles peer-reviewed research studies and scholarly articles that explore the applications, methodologies, and advancements of PCR in equine science.
Cohen ND, Kahn SK, Bordin AI, Gonzales GM, da Silveira BP, Bray JM, Legere RM, Ramirez-Cortez SC.Intragastric administration of virulent Rhodococcus equi protects foals against subsequent experimental intrabronchial (IB) infection, but it is unknown whether R. equi naturally ingested by foals contributes to their susceptibility to pneumonia. Objective: Fecal concentration of virulent R. equi before IB infection with R. equi is positively associated with protection from pneumonia in foals. Methods: Twenty-one university-owned foals. Methods: Samples were collected from experimental studies. Five foals were gavaged with live, virulent R. equi (LVRE) at age 2 and 4 days; the remaining 16 f...
Equine piroplasmosis is a disease of equids, caused by tick-borne apicomplexan protozoan pathogens Babesia caballi and Theileria equi, which, according to the World Organisation for Animal Health (OIE), can be diagnosed by enzyme-linked immunosorbent assay (ELISA), immunofluorescent antibody test (IFAT) and polymerase chain reaction (PCR). The present study was conducted to evaluate and compare the assays available for the diagnosis of equine piroplasmosis. Data employed were obtained from 1300 blood samples collected between 2012-2014 from asymptomatic and symptomatic equines (horses and donk...
Tong P, Duan R, Palidan N, Deng H, Duan L, Ren M, Song X, Jia C, Tian S, Yang E, Kuang L, Xie J.EHV-1 is one of the most serious viral pathogens that frequently cause abortion in horses around the world. However, so far, relatively little information is available on EHV-1 infections as they occur in China. In January 2021, during an abortion storm which occurred in Yili horses at the Chinese State Studs of Zhaosu (North Xinjiang, China), 43 out of 800 pregnant mares aborted. Results: PCR detection revealed the presence of EHV-1 in all samples as the possible cause of all abortions, although EHV-4, EHV-2 and EHV-5 were also found to circulate in the aborted fetuses. Furthermore, the parti...
Hong JH, Oh CS, Kim S, Kang IU, Shin DH.To understand the domestication and spread of horses in history, genetic information is essential. However, mitogenetic traits of ancient or medieval horses have yet to be comprehensively revealed, especially for East Asia. This study thus set out to reveal the maternal lineage of skeletal horse remains retrieved from a 15th century archaeological site (Gongpyeongdong) at Old Seoul City in South Korea. Methods: We extracted DNA from the femur of Equus caballus (SNU-A001) from Joseon period Gongpyeongdong site. Mitochondrial (mt) DNA (HRS 15128-16116) of E. caballus was amplified by polymerase ...
Falcão MVD, Laroucau K, Vorimore F, Deshayes T, Santana VLA, Silva KPC, do Nascimento SA, de Castro RS, Araújo FR, Mota RA.Glanders is an infectious zoonosis caused by Burkholderia (B.) mallei that mainly affects equids. The objective of this work was to provide additional knowledge on the diversity of the strains circulating in Brazil. Six Burkholderia mallei isolates obtained during necropsies of glanderous horses between 2014 and 2017 in two different states (Pernambuco and Alagoas) were analyzed by polymerase chain reaction-high-resolution melting (PCR-HRM). While four strains (9902 RSC, BM_campo 1, BM_campo 3 and UFAL2) clustered in the L3B2 branch, which already includes the Brazilian 16-2438_BM#8 strain, tw...
Cabral LDR, Sato AP, de Sousa RS, Rossa AP, Sanches AWD, Bortoleto CT, Locatelli Dittrich R.The closely related apicomplexa protozoa, Toxoplasma gondii, Neospora spp., and Sarcocystis neurona, have all been associated with neurological and reproductive diseases in horses. However, there is limited data regarding the presence of these three parasites in equine placental tissues and amniotic fluid. The aim of the present report was to investigate the presence of the DNA of T. gondii, Neospora spp. and S. neurona in placentas and amniotic fluid in mares. Anti-T. gondii, anti- S. neurona and anti- Neospora spp. antibody titers were evaluated in 31 mares in the final third of pregnancy by...
Jaramillo-Morales C, Gomez DE, Renaud D, Arroyo LG.The objectives of this study were to determine the prevalence of S. equi in a horse population in Colombia, to determine the risk factors associated with its detection in the guttural pouches and to report the antimicrobial susceptibility of the isolates. Fifteen farms and 137 horses >6 months of age were enrolled. Sampling was randomly, stratified and proportional to the population size of each farm. The guttural pouch (GP) was swabbed via endoscopic guidance and culture was performed. DNA extraction and conventional PCR were performed in colonies compatibles with S. equi, the PCR products we...
Addis A, Canciani E, Campagnol M, Colombo M, Frigerio C, Recupero D, Dellavia C, Morroni M.Different xenogeneic inorganic bone substitutes are currently used as bone grafting materials in oral and maxillo-facial surgery. The aim of the present study was to determine the physicochemical properties and the in vivo performance of an anorganic equine bone (AEB) substitute. AEB is manufactured by applying a process involving heating at >300 °C with the aim of removing all the antigens and the organic components. AEB was structurally characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), X-ray fluorescence (XRF), and Fourier-transformed infrared (FT-IR) spectros...
Tozaki T, Ohnuma A, Kikuchi M, Ishige T, Kakoi H, Hirota KI, Kusano K, Nagata SI.Processed pseudogenes, also known as retrocopy genes, are copies of messenger RNAs that have been reverse transcribed into DNA and inserted into the genome. In this study, we identified 62 processed pseudogene candidates as intron-less genes from whole-genome sequencing (WGS) data of Thoroughbred horses using delly structural variation software. The 62 processed pseudogene candidates were confirmed by PCR amplification of intron-less products. A total of 11 processed pseudogenes were confirmed in the genome of all 23 analysed horses, whereas three processed pseudogenes with structures of ATP11...
Maniego J, Pesko B, Hincks P, Taylor P, Stewart G, Proudman C, Scarth J, Ryder E.The misuse of gene therapy by the introduction of transgenes via plasmid or viral vectors as a doping agent is an increasing concern in human and animal sports, not only in consideration to fair competition but also in potential detrimental effects to welfare. Doping events can be detected by polymerase chain reaction (PCR) amplification of a transgene-specific region of DNA. Quantitative real-time PCR (qPCR) is particularly suited to confirmatory investigations where precise limits of detection can be calculated. To fully validate a qPCR experiment, it is highly desirable to confirm the ident...
Yoon J, Park T, Kim A, Song H, Park BJ, Ahn HS, Go HJ, Kim DH, Lee JB, Park SY, Song CS, Lee SW, Choi IS.Equine parvovirus-hepatitis (EqPV-H) and equine hepacivirus (EqHV) are etiologically associated with Theiler's disease (TD), causing fulminant equine hepatitis, but the transmission route and co-infection effect remain unclear. We determined EqPV-H and EqHV prevalence and coinfection rate in 160 serum and 114 faecal samples using nested polymerase chain reaction. Amino acid and nucleotide analyses were performed and phylogenetic trees were constructed. By measuring liver-specific parameters (AST, GGT, TBIL and A/G ratio), hepatopathological changes in viremia status were compared. We found a h...
Tozaki T, Ohnuma A, Kikuchi M, Ishige T, Kakoi H, Hirota KI, Kusano K, Nagata SI.One method of gene doping in horseracing is administering of exogenous genetic materials, known as transgenes. Several polymerase chain reaction (PCR)-based methods have been developed for detecting transgenes with high sensitivity and specificity. However, novel designs for reference materials (RMs) and/or positive template controls (PTCs) are necessary for simultaneous analysis of multiple transgene targets. In this study, we designed and developed a novel RM for simultaneously detecting multiple targets via microfluidic quantitative PCR (MFQPCR). Twelve equine genes were selected as targets...
Abdullah HHAM, Aboelsoued D, Farag TK, Abdel-Shafy S, Abdel Megeed KN, Parola P, Raoult D, Mediannikov O.Equine vector-borne diseases (EVBDs) are emerging and re-emerging diseases, and most of them are zoonotic. This study aimed to investigate EVBDs in equines and associated arthropods (ticks and flies) from Egypt using molecular analyses, in addition to a preliminary characterization of associated ticks and flies by the matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) and molecular techniques. In this study, 335 blood samples were obtained from equines that appeared to be in good health (320 horses and 15 donkeys) in Cairo and Beni Suef provinces, Egypt. From the same anima...
Temesgen T, Getachew Y, Negussie H.Equine herpesvirus (EHV) infections have major economic, health, and welfare impacts on equids. This study was performed in three selected zones of central Ethiopia with the objectives of detecting EHV-1, -2, and -5 in horses and donkeys with suggestive signs of respiratory tract disease and to assess epidemiological risk factors associated with infections. Methods: A total of 58 nasopharyngeal swab samples were collected from donkeys and horses showing clinical signs of respiratory disease. Polymerase chain reaction (PCR) was used to detect EHV-1, -2, and -5. Evaluation of the associated risk...
Kinoshita Y, Kakoi H, Ishige T, Yamanaka T, Niwa H, Uchida-Fujii E, Nukada T, Ueno T.Taylorella equigenitalis causes contagious equine metritis. Here we compared seven nucleic acid amplification tests for T. equigenitalis to select a rapid and reliable diagnostic method. The 95% detection limits of each assay varied greatly: real-time PCR had the lowest detection limit (0.77 fg/reaction); those of some of the conventional PCRs (cPCRs) were >100 fg/reaction. In experimentally infected samples, real-time PCR and semi-nested PCR showed the highest positive numbers (33 out of 42 samples), but two of the cPCRs detected only 2 and 7 positive results. Our results indicate that the us...
Rahmani SS, Malekifard F, Tavassoli M.Neospora caninum is an Apicomplexan parasite that can cause enormous economic losses due to abortions in cattle. The present study investigated the role of Neospora spp. infection in equine abortion in Iranian donkeys using molecular and phylogenetic analyses. Twenty-nine-aborted fetuses and 29 blood samples from their dams were collected from six different regions in the West and Northwest of Iran. They were tested for N. caninum by PCR at the Nc5 locus, followed by sequencing of five of the PCR products. The overall molecular prevalence was 34.5% in blood samples and the prevalence by DNA de...
Risso A, Campos G, Garcia H, Zerpa H.Equine piroplasmosis (EP) is a tick-borne infectious disease highly prevalent in tropical and subtropical regions, such as Venezuela. EP affects wild and domestic equids leading to several clinical presentations, from asymptomatic to severely affected animals. In this study, thirty-three (33) sport horses under regular training activities and from endemic regions of north-central Venezuela were submitted to an observational survey, case-control, to describe the presence of clinical signs and natural EP infections. A conventional PCR assay targeting the SSU rRNA gene revealed EP etiologic agent...
Suganuma K, Acosta TJ, Valinotti MFR, Sanchez AR, Mossaad E, Elata A, Inoue N.Despite the epidemic situation of animal trypanosomosis caused by Trypanosoma evansi, Trypanosoma equiperdum and Trypanosoma vivax in South American countries, there are no reports for the prevalence of animal trypanosomes in Paraguay. In this study, 408 blood samples were obtained from apparently healthy horses from sixteen departments of Paraguay, for routine medical check-up from August to September 2019, and a polymerase chain reaction (PCR)-based cross-sectional study was carried out to identify trypanosome prevalence. The prevalence of Trypanozoon (T. evansi and T. equiperdum) and T. viv...
Rodríguez N, Whitfield-Cargile CM, Chamoun-Emanuelli AM, Hildreth E, Jordan W, Coleman MC.The nasopharyngeal bacterial and fungal microbiota of normal horses and those with nasopharyngeal cicatrix syndrome (NCS) are unknown. Objective: To describe the microbiota from nasopharyngeal washes of healthy horses and of horses acutely affected with NCS. Methods: Twenty-six horses acutely affected with NCS horses and 14 unaffected horses. Methods: Prospective, observational cohort study. Horses were recruited by investigators through personal communications in central Texas. Bacterial (16s RNA) and fungal (internal transcribed spacer) microbiota from nasopharyngeal washes were evaluated. P...
Peano A, Arnoldi S, Čmoková A, Hubka V.This article reports the first verified cases of infection by Trichophyton bullosum in Africa since the description of the fungus, isolated in 1933 from the coat of horses in Tunisia and Mali. We found the fungus in cutaneous samples obtained from donkeys suffering from severe dermatitis with areas of alopecia and scaling in the surroundings of Cairo (Egypt). Fungal elements (arthroconidia and hyphae) were seen at the microscopy of material collected by skin scraping and digested in NaOH. Fungal colonies grown on various culture media were identified through PCR and sequencing of the ITS rDNA ...
Elsawy BSM, Nassar AM, Alzan HF, Bhoora RV, Ozubek S, Mahmoud MS, Kandil OM, Mahdy OA.Equine Piroplasmosis (EP) is an infectious disease caused by the hemoprotozoan parasites , , and the recently identified species . Hereby, we used a multiplex PCR (mPCR) targeting the 18S rRNA gene of and for the simultaneous detection of EP in Egyptian equids and examined the presence of infections in Egypt. Blood samples from 155 equids (79 horses and 76 donkeys) collected from different governorates of Egypt were examined by mPCR and PCR targeting . The mPCR method revealed a prevalence of of 20.3% in horses and of 13.1% in donkeys and a prevalence of . of 1.2% in horses. . was not de...
Otzdorff C, Beckmann J, Goehring LS.(1) Background: Equine arteritis virus (EAV) infection causes reproductive losses and systemic vasculitis in susceptible equidae. The intact male becomes the virus' reservoir upon EAV infection, as it causes a chronic-persistent infection of the accessory sex glands. Infected semen is the main source of virus transmission. (2) Here, we describe acute EAV infection and spread in a stallion population after introduction of new members to the group. (3) Conclusions: acute clinical signs, acute phase detection of antigen via (PCR) nasal swabs or (EDTA) blood, and seroconversion support the idea of...
Marčeková P, Mad'ar M, Styková E, Kačírová J, Sondorová M, Mudroň P, Žert Z.Equine hoof canker and bovine digital dermatitis are infectious inflammatory diseases of the hooves with an unknown etiology. However, anaerobic spirochetes of the genus Treponema are considered to be potential etiological agents. The aim of this study was to find a suitable way to isolate DNA and to detect the presence of treponemal DNA in samples of equine hoof canker and bovine digital dermatitis. DNAzol®® Direct and column kits were used to isolate DNA from samples of equine hoof canker and bovine digital dermatitis. The presence of Treponema spp. was detected using PCR and Sanger sequen...
Takai S, Sudo M, Sakai M, Suzuki K, Sasaki Y, Kakuda T, Suzuki Y.Rhodococcus equi was isolated from the gastrointestinal contents of earthworms (family Megascolecidae) and their surrounding soil collected from pastures of two horse-breeding farms in Aomori Prefecture, outdoor pig pens, forest in Towada campus, orange groves and forest where wild boars (Sus scrofa) are established in Tanabe, Wakayama Prefecture. The number of R. equi in the lower gastrointestinal contents of 23 earthworms collected from our campus was significantly larger than that of the upper gastrointestinal content. The mean numbers of R. equi from the gastrointestinal contents of earthw...
Tozaki T, Ohnuma A, Hamilton NA, Kikuchi M, Ishige T, Kakoi H, Hirota KI, Kusano K, Nagata SI.Gene doping is prohibited for fair competition in human and horse sports. One style of gene doping is the administration of an exogeneous gene, called a transgene, to postnatal humans and horses. Although many transgene detection methods based on quantitative polymerase chain reaction (PCR), including real-time PCR and digital PCR, have been recently developed, it remains difficult to reliably detect low-copy transgenes. In this study, we developed and validated a nested digital PCR method to specifically detect low-copy transgenes. The nested digital PCR consists of (1) preamplification using...
Costa VMD, Cursino AE, Franco Luiz APM, Braz GF, Cavalcante PH, Souza CA, Simplício KMMG, Drumond BP, Lima MT, Teixeira BM, Kroon EG.Equine infectious anemia (EIA) is listed by the World Organization for Animal Health (OIE) as one of the equine diseases that must be notified. No effective treatment or vaccine is available. EIA control is based on segregation and euthanasia of positive equids. The disease is caused by the equine infectious anemia virus (EIAV), a member of the genus Lentivirus of the Retroviridae family. Despite the importance of this disease in equids, EIA has been poorly studied in donkeys (Equus asinus). We evaluate the sanitary conditions related to EIAV in donkeys from a shelter of abandoned animals capt...
Pradella GD, Duarte CA, Escobar TA, Zuravski L, Góss GC, Skupien JA, Lübeck I.The Leishmaniases are on the top of the global list of tropical neglected diseases. The number of infected dogs in South America is estimated in millions and correlated to disease cases in humans, especially in Brazil. Equines may get infected too and can play a role in the epidemiological chain. Thus, the aim of the present study was to evaluate risk and protective factors of leishmaniasis in rural areas of the western border region of Rio Grande do Sul state, Brazil by Leishmania spp. protozoa molecular detection and serological evaluation (ELISA) in equine and canine blood samples. This wor...
Savage VL, Christley R, Pinchbeck G, Morrison LJ, Hodgkinson J, Peachey LE.Trypanosomosis is a major cause of morbidity and mortality in working equids in The Gambia. Recently, a progressive, severe neurological syndrome characterised by a diffuse lymphoplasmacytic meningoencephalitis has been identified and associated with Trypanosoma brucei infection of the central nervous system. The pathogenesis of cerebral trypanosomosis is unclear and the clinical syndrome not well described. This observational cross-sectional study aimed to identify host and parasite related risk factors associated with the development of cerebral trypanosomosis and to describe the neurologica...
Munday JS, Gedye K, Daudt C, Chaves Da Silva F.Bovine papillomavirus (BPV) types 1 and 2 are recognized as the main cause of equine sarcoids. However, some studies report that up to a quarter of these tumors do not contain detectible BPV1 or BPV2 DNA. The absence of detectible BPV1 or BPV2 in these sarcoids suggests the possible involvement of other papillomavirus types. Currently, five are recognized to cause mesenchymal neoplasia after cross-species infection. In addition to BPV1 and BPV2, BPV13 has been associated with equine sarcoids in Brazil, BPV14 has been associated with feline sarcoids, and Ovis aries papillomavirus 2 caused a sa...
Yoon J, Park T, Kim A, Park J, Park BJ, Ahn HS, Go HJ, Kim DH, Jung S, Seo Y, Lee JB, Park SY, Song CS, Lee SW, Choi IS.Equine parvovirus-hepatitis (EqPV-H) is a newly identified etiologic agent of Theiler's disease (TD). We present a case of EqPV-H-related fulminant hepatitis in a 14-year-old thoroughbred mare in Korea. The mare had acute hepatopathy and gastrointestinal symptoms, with abnormal liver-related blood parameters. The horse was born in the USA and imported to Korea in 2017, with no history of administration of equine biological products after entry into Korea. The horse was diagnosed with EqPV-H-associated hepatitis after abdominal ultrasonography, laparotomy, and nested polymerase chain reaction (...
Butler CM, Sloet van Oldruitenborgh-Oosterbaan MM, Stout TA, van der Kolk JH, Wollenberg Lv, Nielen M, Jongejan F, Werners AH, Houwers DJ.Equine piroplasmosis (EP) has not been considered indigenous in The Netherlands. However, following the detection of an apparently indigenous subclinical Babesia caballi infection in a horse on Schouwen-Duiveland (an island in the Zeeland Province), a survey was undertaken between May and September 2010 to assess the prevalence of the causative agents of EP in the South-West of The Netherlands. Blood samples from 300 randomly selected horses were tested for specific antibodies against Theileria equi and B. caballi using an indirect fluorescence antibody test (IFAT), and for parasite DNA using ...
Traversa D, Giangaspero A, Galli P, Paoletti B, Otranto D, Gasser RB.Gastric or cutaneous habronemosis caused by Habronema microstoma Creplin, 1849 and Habronema muscae Carter, 1865 is a parasitic disease of equids transmitted by muscid flies. There is a paucity of information on the epidemiology of this disease, which is mainly due to limitations with diagnosis in the live animal and with the identification of the parasites in the intermediate hosts. To overcome such limitations, a molecular approach, based on the use of genetic markers in the second internal transcribed spacer (ITS-2) of ribosomal DNA, was established for the two species of Habronema. Charact...
Carossino M, Lee PY, Nam B, Skillman A, Shuck KM, Timoney PJ, Tsai YL, Ma LJ, Chang HF, Wang HT, Balasuriya UB.Equine arteritis virus (EAV) is the causative agent of equine viral arteritis (EVA), a respiratory and reproductive disease of horses. Most importantly, EAV induces abortion in pregnant mares and can establish persistent infection in up to 10-70% of the infected stallions, which will continue to shed the virus in their semen. The objective of this study was to develop and evaluate a reverse transcription insulated isothermal polymerase chain reaction (RT-iiPCR) for the detection of EAV in semen and tissue samples. The newly developed assay had a limit of detection of 10 RNA copies and a 10-fol...
Tozaki T, Gamo S, Takasu M, Kikuchi M, Kakoi H, Hirota KI, Kusano K, Nagata SI.Doping control is an important and indispensable aspect of fair horse racing; genetic doping has been recently included to this. In this study, we aimed to develop a detection method of gene doping. A plasmid cloned with human erythropoietin gene (p.hEPO, 250 μg/head) was intramuscularly injected into a microminipig. Subsequently, p.hEPO was extracted from 1 mL of plasma and detected by droplet digital polymerase chain reaction. Results: The results confirmed that the maximum amount of plasmid was detected at 15 min after administration and the majority of the plasmid was degraded in the b...
Cohen ND, Martin LJ, Simpson RB, Wallis DE, Neibergs HL.To compare the sensitivity of polymerase chain reaction (PCR) with microbiological culture for detecting salmonellae in equine fecal samples and equine environmental swab specimens. Methods: Samples and specimens were tested by PCR and microbiological culture. Methods: A fecal sample from each of 152 horses admitted consecutively to the clinic for evaluation by the outpatient service, 282 fecal samples from 110 hospitalized horses that had been submitted to the clinical microbiology laboratory, and 313 environmental swab specimens were examined. Methods: Each sample and specimen in the study w...
Peat FJ, Colbath AC, Bentsen LM, Goodrich LR, King MR.This study aimed to examine the influence of neodymium-doped yttrium aluminum garnet (Nd:YAG) laser irradiation on equine bone marrow-derived mesenchymal stem cell (MSC) viability, proliferation, and cytokine expression in vitro. Background: Photobiomodulation of cells using monochromatic light is a technique designed to influence cellular processes. Previous studies have shown dose-dependent effects of low-level laser irradiation on cell proliferation and cytokine expression in a range of cell types and species. Evidence for the influence of 1064 nm wavelength near-infrared irradiation on M...
Abedi V, Razmi G, Seifi H, Naghibi A.Equine piroplasmosis is a hemoprotozoan tick-borne disease with worldwide distribution that is caused by Theileria equi and Babesia caballi. However, the geographical distribution of equine piroplasmosis in Iran is unknown. The aim of the current study was to determine the causative agents and vector ticks of equine piroplasmosis in horses in the North Khorasan Province. In the year 2011, 100 horses were randomly selected from 14 villages. Blood samples and ixodid ticks were collected and examined using microscopical, molecular, and serological methods. Theileria equi infection was microscopic...
Fortier G, van Erck E, Fortier C, Richard E, Pottier D, Pronost S, Miszczak F, Thiry E, Lekeux P.The objectives of this study were to estimate the prevalence and the potential role of equine herpesviruses (EHVs) detection in both bronchoalveolar lavage (BAL) and tracheal wash (TW). The population included a control group (CTL; 37 TW and 25 BAL) and a pathological group (PAT; 259 TW and 387 BAL), including horses either suffering from respiratory diseases including syndrome of tracheal inflammation, inflammatory airway disease, recurrent airway obstruction, or submitted to respiratory investigation because of exercise intolerance or poor performance. Each respiratory liquid was submitted t...
Hastie PM, Mitchell K, Murray JA.There is a need to further our understanding of the role that the equine hindgut ecosystem plays in digestive processes and diseases. The aim of the present study was to utilise the real-time PCR technique to determine the abundance of candidate cellulolytic (Ruminococcus flavefaciens; Fibrobacter succinogenes) and non-cellulolytic (Streptococcus bovis) bacteria in lumen contents from the caecum, ventral and dorsal colon, and rectum of healthy horses (n 14). Total DNA was extracted from frozen and lyophilised lumen contents, and PCR primers and Taqman probes were designed based on 16S rDNA seq...
Elhaig MM, Sallam NH.In Egypt, although the Trypanosoma evansi has been reported frequently among domestic animals, there is no published data on T. evansi in horses. Therefore, this study aimed to assess the prevalence and characterization of T. evansi in three governorates by examining blood samples from 40 local camels, 35 imported camels, 25 horses and 10 donkeys by PCR targeting the sequences of TBR and RoTat 1.2VSG. The overall prevalence of T. evansi was 54.5% and 21.8% by TBR PCR and RoTat 1.2VSG PCR, respectively. The TBR PCR detected T. evansi in 60% and 71.4%, respectively, of local and imported camels ...
Grandi G, Molinari G, Tittarelli M, Sassera D, Kramer LH.Babesia caballi and Theileria equi are the causative agents of equine piroplasmosis. In this epidemiological study, 294 horses reared in a rural area of northern Italy were studied. During January 2008-January 2009, blood samples were taken for serology (indirect fluorescent antibody test) and for polymerase chain reaction (PCR). Data on the geographical area, sex, and age were collected for statistical analysis of risk factors associated with infection. A seroprevalence of 8.5% was found: 8.2% of the animals were positive for anti-T. equi antibodies and 0.3% for anti-B. caballi antibodies. No...
Fehr JE, Trotter GW, Oxford JT, Hart DA.To determine relative amounts of mRNA expression of aggrecan, type-II collagen, matrix metalloproteinase (MMP) 1, and MMP3 in articular cartilage and synovial membrane samples from healthy equine joints and joints with osteoarthritis (OA) and to compare results of Northern blot hybridization with results of a reverse transcriptase-polymerase chain reaction (RT-PCR) assay. Methods: Articular cartilage samples from 8 pairs of joints (1 with OA and 1 healthy) from 6 horses and synovial membrane samples from 6 pairs of joints from 5 horses. Methods: RNA was extracted from samples by use of a modif...
Liu A, Zhang J, Zhao J, Zhao W, Wang R, Zhang L.Horses interact with humans in a wide variety of sport competitions and non-competitive recreational pursuits as well as in working activities. Cryptosporidium spp are one of the most important zoonotic pathogens causing diarrhea of humans and animals. The reports of Cryptosporidium in horses and the findings of zoonotic Cryptosporidium species/genotypes show a necessity to carry out molecular identification of Cryptosporidium in horses, especially in diarrheic ones. The aim of the present study was to understand Cryptosporidium infection and species/genotypes in diarrheic horses, and to trace...
Vo AT, van Duijkeren E, Fluit AC, Gaastra W.To investigate the genotypic resistance of integron-carrying Salmonella Typhimurium isolates from horses and their genetic relationship. Methods: Sixty-one Salmonella isolates were screened for the presence of class 1 integrons by PCR. The gene cassettes of integron-positive isolates were detected by PCR, restriction fragment length polymorphism typing, and sequencing. The potential for the transfer of resistance determinants was investigated by conjugation experiments. The presence of Salmonella genomic island 1 (SGI1) or its variants was studied by PCR and nucleotide sequencing. PFGE was use...
Bera BC, Virmani N, Kumar N, Anand T, Pavulraj S, Rash A, Elton D, Rash N, Bhatia S, Sood R, Singh RK, Tripathi BN.Equine influenza is a major health problem of equines worldwide. The polymerase genes of influenza virus have key roles in virus replication, transcription, transmission between hosts and pathogenesis. Hence, the comprehensive genetic and codon usage bias of polymerase genes of equine influenza virus (EIV) were analyzed to elucidate the genetic and evolutionary relationships in a novel perspective. Results: The group - specific consensus amino acid substitutions were identified in all polymerase genes of EIVs that led to divergence of EIVs into various clades. The consistent amino acid changes...
Brum MC, Anjos BL, Nogueira CE, Amaral LA, Weiblen R, Flores EF.An outbreak of severe cutaneous disease associated with an orthopoxvirus in horses in southern Brazil is described. Fourteen Crioulo mares and foals from a husbandry farm developed papules, and vesicles progressing to proliferative and exudative lesions on the muzzle, external nares, and external and internal lips. The vesicles eroded, and the proliferative lesions eventually bled and progressed to moist crusts and scars. The clinical signs lasted approximately 6-12 days, after which the animals progressively recovered. Direct electron microscopy of skin biopsies revealed brick-shaped, 250-300...
Pagamjav O, Sakata T, Matsumura T, Yamaguchi T, Fukushi H.Equine herpesvirus 1 (EHV-1) is a pathogen causing rhinopneumonia in young horses, abortion in mares, and myeloencephalitis in adult horses. Two types, EHV-1 P and EHV-1 B, have recently been dominant among 16 electropherotypes. EHV-1 P and EHV-1 B viruses were compared by long and accurate polymerase chain reaction (LA-PCR) and restriction fragment length polymorphism (RFLP) analysis. Differences in restriction sites were found to be focused in ORF64, which encodes the infected cell protein 4 (ICP4), and downstream of the ICP4 gene. The 3 ' -end and downstream of ICP4 gene of EHV-1 B were fou...
Léon A, Fortier G, Fortier C, Freymuth F, Tapprest J, Leclercq R, Pronost S.The major role of EHV-1 in equine abortion is widely reported in the literature but the contribution of EHV-2, EHV-3, EHV-4 or EHV-5 remains less well documented. The objective of this study is to evaluate the contribution of these five different EHVs to equine abortion in a variety of biological tissues using a consensus polymerase chain reaction (PCR). The test was validated for specificity and sensitivity in horses before screening specimens from 407 foetuses, stillbirths and premature foals collected over a 2.5-year interval. Positive results obtained with this assay were compared to other...
Lei R, Wang X, Zhang D, Liu Y, Chen Q, Jiang N.Equine piroplasmosis (EP) is a severe disease of horses caused by the tick-borne protozoa Theileria equi (T. equi) and Babesia caballi (B. caballi). Infectious carriers are not always symptomatic, meaning there is a risk to non-enzootic areas. Regulatory tests for EP include sero-epidemiological methods for equine babesiosis, but these lack specificity due to cross-reactivity with other Babesia species. In this study, we present a real-time quantitative recombinase polymerase amplification (qRPA) method for fast simultaneous detection of both T. equi and B. caballi. In this method, primers and...
Arroyo LG, Staempfli H, Weese JS.Clostridium difficile is an important cause of diarrhea in horses, causing sporadic and epidemic disease of varying severity. This study evaluated the molecular characteristics of 48 C. difficile isolates recovered from diarrheic horses admitted to a veterinary hospital by using PCR-ribotyping and toxin gene profile. Additionally, feces were tested for the presence of C. difficile toxin A/B via enzyme immunosorbant assay (EIA) in 38 horses. The toxin genes tcdA, tcdB and cdtB were present in 27 (56.25%), 35 (72.91%) and 2 (4.1%) strains, respectively. Eight isolates (16.6%) were A(-)B(+) varia...
Sloboda M, Jirků M, Lukešová D, Qablan M, Batsukh Z, Fiala I, Hořín P, Modrý D, Lukeš J.Equine piroplasmosis caused by Babesia caballi and Theileria equi is widespread in Asia. The presence of these haemozoans in Mongolia was previously confirmed in domestic as well as in reintroduced Przewalski horses in which they cause significant pathology. The data on occurrence of piroplasms from Bactrian camels in Asia is lacking. A total of 192 horses, 70 Bactrian camels, and additional 16 shepherd dogs from the Hentiy province were included in our study. No clinical signs typical for piroplasmid infection were observed during the field survey. Microscopic examination revealed the presenc...
Burton AJ, Nydam DV, Dearen TK, Mitchell K, Bowman DD, Xiao L.To date, little is known about the prevalence, genotypes and zoonotic potential of Cryptosporidium spp. affecting horses, especially in North America. A cross-sectional study was conducted in New York, USA between February 25th and May 1st 2009. Fecal samples were collected from three hundred and forty nine 1-10-week-old foals and their dams on 14 different broodmare farms. All fecal samples were screened for Cryptosporidium spp. using a direct immunofluorescence assay (DFA). DNA extraction and PCR-RFLP analysis of the small-subunit (SSU) rRNA gene were performed on all the foal samples. PCR-p...
Clausen PH, Chuluun S, Sodnomdarjaa R, Greiner M, Noeckler K, Staak C, Zessin KH, Schein E.From May to July 2000, a cross-sectional study was conducted to estimate the prevalence of Trypanosoma equiperdum in the horse population of the central province (Tuv aimag) of Mongolia. On average, four herds were selected from each of the 29 aimag subdivisions (119 herds). From each herd, 10 horses were sampled in proportion to sex and age categories in the respective herds (1190 horses). Sera from 1122 horses were analysed for T. equiperdum antibodies using two serological assays, the complement fixation test (CFT) and the enzyme-linked immunosorbent assay (ELISA). The crude estimate of the...
Balasuriya UB, Crossley BM, Timoney PJ.Equid herpesvirus 1 (EHV-1) is one of the most economically important equine viral pathogens. Its clinical manifestations in horses vary from acute upper respiratory tract disease, abortion, or neonatal death, to neurological disease termed equine herpesviral myeloencephalopathy, which may lead to paralysis and a fatal outcome. Successful identification of EHV-1 infection in horses depends on a variety of factors such as suitable case selection with emphasis on timing of sample collection, selection of appropriate sample(s) based on the clinical manifestations, application of relevant diagnost...
Nyaoke AC, Navarro MA, Beingesser J, Uzal FA.A 14-y-old bay Quarter Horse gelding was presented with progressive neurologic signs, elevated rectal temperature, and icterus for 3 d prior to death. Postmortem examination revealed icterus, large amounts of serosanguineous fluid in the abdominal cavity, widespread petechiae and ecchymoses in several organs, and a large, pale, and well-demarcated focus of necrosis in the liver. Histologically, there was coagulative necrosis surrounded by a rim of inflammatory cells and large numbers of gram-positive rods, which were identified as Clostridium novyi by immunohistochemistry. Liver samples tested...
Laroucau K, Aaziz R, Vorimore F, Varghese K, Deshayes T, Bertin C, Delannoy S, Sami AM, Al Batel M, El Shorbagy M, Almutawaa KAW, Alanezi SJ....Glanders is a contagious zoonotic disease caused by Burkholderia mallei. Following the detection of glanders positive horses using the OIE complement fixation test, the tissues of two horses were analysed by PCR. While PCR systems targeting the Burkholderia pseudomallei complex gave positive signals, the species-specific PCR systems targeting B. mallei (fliP-IS407A) and B. pseudomallei (orf11)-the OIE recommended targets-resulted in negative signals. However, the presence of B. mallei in these tissues was confirmed with a recently described B. mallei-specific real-time PCR system and genot...
Lu Z, Branscum AJ, Shuck KM, Zhang J, Dubovi EJ, Timoney PJ, Balasuriya UB.Two previously developed TaqMan fluorogenic probe-based 1-tube real-time reverse transcription polymerase chain reaction (real-time RT-PCR) assays (T1 and T2) were compared and validated for the detection of Equine arteritis virus (EAV) nucleic acid in equine semen and tissue culture fluid (TCF). The specificity and sensitivity of these 2 molecular-based assays were compared to traditional virus isolation (VI) in cell culture. The T1 real-time RT-PCR had a higher sensitivity (93.4%) than the T2 real-time RT-PCR (42.6%) for detection of EAV RNA in semen. However, the T1 real-time RT-PCR was les...
Gallusová M, Qablan MA, D'Amico G, Oborník M, Petrželková KJ, Mihalca AD, Modrý D.Rural areas of Romania, particularly the localities covering Danube Delta, are still not sufficiently explored in terms of epidemiological aspects, despite the large density of domestic animals living in close contact with people and natural environment of the Danube Delta Biosphere Reserve. Between 2010 and 2012, a survey on equine piroplasmids species was performed in this area, using a total of 178 horses, 15 donkeys and 177 dogs from 18 localities inside and outside the Danube Delta. None of the sampled hosts showed any clinical symptoms typical for equine piroplasmoses. A 25.4% overall pr...
Berlin D, Loeb E, Baneth G.Trypanosomiasis caused by Trypanosoma evansi ("Surra") is mainly a wasting disease affecting equids, camels and cattle as well as other domestic and wild animal species. In horses, infection may cause severe neurological abnormalities; however, the clinical progression, pathogenesis and molecular ante-mortem detection of this form of the disease have not been described in detail. A mare with progressive ataxia, head tilt, nystagmus and cranial nerve deficits submitted to treatment was diagnosed with central nervous system trypanosomiasis following the detection of a Trypanosoma tryposmastigote...
Hedberg-Alm Y, Penell J, Riihimäki M, Osterman-Lind E, Nielsen MK, Tydén E.All grazing horses are exposed to intestinal parasites, which have the potential to cause gastrointestinal disease. In Sweden, there is a concern about an increase in parasite-related equine gastrointestinal disease, in particular , since the implementation of prescription-only anthelmintics approximately 10 years ago. In a prospective case-control study, parasitological status, using fecal analyses for strongyle egg counts, the presence of eggs and Polymerase chain reaction (PCR) as well as serology for , were compared between horses presenting with or without gastrointestinal disease at a ...
