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.
Borchers K, Ebert M, Fetsch A, Hammond T, Sterner-Kock A.Equine herpes virus 2 (EHV-2), a gamma(2)-herpesvirus, is common in horses of all ages. Its role as a primary pathogen is unclear but there is an association between EHV-2, respiratory disease and keratoconjunctivitis. The purpose of this study was to gain more information on the prevalence of EHV-2 DNA in conjunctival swabs from horses with and without ocular disease and to define the anatomical site and cell type harbouring viral genome or antigen. By polymerase chain reaction (PCR) 22 out of 77 (28.6%) ocular swabs of clinically healthy and only 4 out of 48 (8.3%) samples from diseased hors...
Journal of medical entomologyAugust 9, 2006
Volume 43, Issue 4 762-773 doi: 10.1603/0022-2585(2006)43[762:DACOBB]2.0.CO;2
Morshed MG, Scott JD, Fernando K, Geddes G, McNabb A, Mak S, Durden LA.The blacklegged tick, Ixodes scapularis Say (Acari: Ixodidae), has a wide geographical distribution in Ontario, Canada, with a detected range extending at least as far north as the 50th parallel. Our data of 591 adult I. scapularis submissions collected from domestic animals (canines, felines, and equines) and humans during a 10-yr period (1993-2002) discloses a monthly questing activity in Ontario that peaks in May and October. The Lyme disease spirochete Borrelia burgdorferi Johnson, Schmidt, Hyde, Steigerwalt & Brenner was detected in 12.9% of I. scapularis adults collected from domesti...
Takai S, Zhuang D, Huo XW, Madarame H, Gao MH, Tan ZT, Gao SC, Yan LJ, Guo CM, Zhou XF, Hatori F, Sasaki Y, Kakuda T, Tsubaki S.Little is known about the distribution of Rhodococcus equi in the soil environment of native horses in China. One hundred and eight soil samples were collected from native-horse farms in the Hulun Beier grasslands of eastern Mongolia, the Xilin Goler grasslands of southern Mongolia, and Tongliao City in Inner Mongolia, China. The isolation rates of R. equi from soil samples from the Hulun Beier and Xilin Goler grasslands ranged from 25.9% to 30.0%. In contrast, isolation rates from soil samples from Tongliao City were as high as 82.3% and the mean number of R. equi in soil samples from Tonglia...
Allen GP.To evaluate a technique for identifying horses latently infected with neuropathogenic strains of equine herpesvirus-1 (EHV-1). Methods: 36 adult mares, 24 of which were experimentally infected as weanlings with neuropathogenic or nonneuropathogenic EHV-1. Methods: Mandibular lymph node (MLN) tissue was obtained from each horse via biopsy during general anesthesia. Purified DNA from MLNs was tested for EHV-1 DNA by use of a magnetic bead, sequencecapture, nested PCR assay. For MLNs that contained EHV-1 DNA, the 256-bp DNA fragments amplified via sequence-capture nested PCR were sequenced to det...
Li JL, Shi YF, Bu RQ, Mang L.Restriction endonucleases, namely BamH I, Taq I, Hae III, Rsa I and Hinc II, were used to analyze the polymorphism of partial mtDNA Cytb gene sequences from 256 horses 6 types (Thoroughbred, Sanhe, Wuzhumuqin, Xinihe, Wushen and Pony) including the imported breed, cultivated breed and local breed. The products of endonuclease digestion were run on 8% non-denaturing polyacrylamide gel electrophoresis and detected by silver staining. Results indicated BamH I and Taq I polymorphism. In all 7 restriction patterns were defected that could be sorted into 3 haplotypes, of which haplotypes I and III w...
Sledge DG, Miller DL, Styer EL, Hydrick HA, Baldwin CA.Granulomatous dermatitis in horses has been linked to many etiologies, including various parasites, fungi, and bacteria. Idiopathic forms of granulomatous inflammation-producing diseases, some of which are localized to the skin, also have been reported in horses. Herein we describe a case of recurrent equine granulomatous skin disease characterized by intranuclear viral inclusions within macrophages and giant cells. The histologic changes were primarily noted in the deep dermis and included multifocal to coalescing areas of necrosis marked by histiocytic cell infiltration and presence of giant...
Hornyák A, Bakonyi T, Kulik M, Kecskeméti S, Rusvai M.The occurrence of two important pathogens, equine herpesvirus 1 (EHV1) and equine arteritis virus (EAV) causing abortions, perinatal foal mortality and respiratory disease, was investigated by polymerase chain reaction (PCR) and virus isolation to demonstrate the presence of abortigenic viruses in samples from 248 horse fetuses in Hungary. We found 26 EHV1- and 4 EAV-positive aborted or prematurely born foals from 16 and 4 outbreaks, respectively, proving that despite the widely applied vaccination, EHV1 is a far more important cause of abortions in the studs than EAV. We compared the virus co...
Donnelly BP, Nixon AJ, Haupt JL, Dahlgren LA.To characterize the nucleotide sequence of equine platelet-derived growth factor (PDGF)-A and -B and analyze temporal expression of these genes in equine tendon after induced tendinitis injury. Animals-18 mature horses. Methods: Genes for equine PDGF-A and -B were reverse transcribed and sequenced from synovial tissue mRNA obtained from a 3-year-old horse. Collagenase-induced lesions were created in the tensile region of the superficial digital flexor tendon in 14 horses; 3 horses served as uninjured control animals. Tendons were harvested and total RNA was isolated from experimental horses 1,...
Duquesne F, Pronost S, Laugier C, Petry S.A direct-PCR assay was developed for the rapid detection of Taylorella equigenitalis, a Gram-negative bacterium responsible for contagious equine metritis (CEM) in Equidae. The bacteria may be detected in equine genital swabs without need for a preliminary step of DNA extraction or bacterial isolation. Specificity was determined with 125 isolates of T. equigenitalis, 24 isolates of Taylorella asinigenitalis, five commensal bacteria of the genital tract and a facultative intracellular pathogen of foals found in large concentration in soil. Our PCR is specific and amplified a 413-bp 16S ribosoma...
Traversa D, Iorio R, Capelli G, Paoletti B, Bartolini R, Otranto D, Giangaspero A.Gastric habronemosis of horses caused by Habronema microstoma and Habronema muscae (Nematoda, Spirurida) is characterized by catarrhal gastritis, diarrhoea, progressive weight loss and ulcers. Despite its importance in the equine industry and in clinical practice, knowledge of the epidemiology of this infection is still incomplete as diagnosis in live animals is challenging. A two-step semi-nested PCR assay using ribosomal DNA (rDNA) markers has recently been used for the molecular diagnosis in vivo of gastric habronemosis based on the detection of H. microstoma and/or H. muscae DNA in equine ...
Sato T, Sato G, Shoji Y, Itou T, Sakai T.After RNA extraction from horsehair shafts and roots, the mRNAs of beta-actin, muscle-type phosphofructokinase, and transforming growth factor-beta1 were detected by reverse transcription polymerase chain reaction assay. Low amounts of RNA were present in the horsehair. These specific mRNA transcripts were readily detected when more than three hair roots were used. However, detection of the mRNA transcripts was difficult in the hair shaft. These findings indicate that the small amounts of residual RNA in horsehair roots can be utilized as samples for molecular biological analysis.
Kim YK, Noh KB, Han CS, Moon JY, Yoon DK, Song KJ, Kim DJ, Kubera M, Maes M, Song JW.Borna disease virus (BDV) predominantly infects horses and sheep, causing a broad range of behavioural disorders. It is controversial whether BDV infects humans and causes psychiatric disorders. Objective: We searched for BDV-derived nucleic acids in blood of race horses and jockeys riding the horses. Methods: We assayed for the BDV genome in RNA extracted from peripheral blood mononuclear cells (PBMC) of 39 race horses and 48 jockeys. Two polymerase chain reaction protocols [one-tube reverse transcription-polymerase chain reaction (RT-PCR) and two-step RT-PCR] were used to assay BDV p24 and p...
Morrell JM, Geraghty RM.A method of removing equine arteritis virus (EAV) from equine semen used for artificial insemination is urgently needed. Recent medical studies suggest that a double semen processing technique of density gradient centrifugation followed by a 'swim-up' can provide virus-free sperm preparations for assisted reproduction. Objective: To investigate the use of the double semen processing technique to obtain virus-free sperm preparations from stallion semen containing EAV. Methods: Aliquots of an ejaculate from an uninfected stallion were spiked with virus and processed by the double processing tech...
Vázquez-Morón S, Avellón A, Echevarría JE.The Lyssavirus genus includes seven species or genotypes named 1-7. Rabies genotypes correlate with geographical distribution and specific hosts. Co-circulation of different lyssaviruses, imported cases, and the presence of unknown viruses, such as Aravan, Khujand, Irkut and West Caucasian Bat Virus, make it necessary to use generic methods able to detect all lyssaviruses. Primer sequences were chosen from conserved regions in all genotypes in order to optimise a generic RT-PCR. Serial dilutions of 12 RNA extracts from all seven Lyssavirus genotypes were examined to compare the sensitivity of ...
Loftin KC, Diallo AA, Herbert MW, Phaltankar PG, Yuan C, Grefe N, Flemming A, Foley K, Williams J, Fisher SL, Elberfeld M, Constantine J, Burcham M....To investigate the occurrence of West Nile virus (WNV) and Eastern equine encephalitis virus (EEE) in southeastern Virginia, the Bureau of Laboratories at the Norfolk Department of Public Health (NDPH) analyzed mosquito pools and the sera of sentinel chickens from the southeastern Virginia area each year from 2000 to 2004. Mosquito pool supernatants were screened for the presence of viral RNA by conventional reverse transcription polymerase chain reaction (RT-PCR) and Taqman RT-PCR with the i-Cycler. Mosquito pools were also tested for virus activity by Vero cell culture. The primary enzootic ...
Bogaert L, Van Poucke M, De Baere C, Peelman L, Gasthuys F, Martens A.Real-time quantitative PCR can be a very powerful and accurate technique to examine gene transcription patterns in different biological conditions. One of the critical steps in comparing transcription profiles is accurate normalisation. In most of the studies published on real-time PCR in horses, normalisation occurred against only one reference gene, usually GAPDH or ACTB, without validation of its expression stability. This might result in unreliable conclusions, because it has been demonstrated that the expression levels of so called "housekeeping genes" may vary considerably in different t...
Mittelholzer C, Stadejek T, Johansson I, Baule C, Ciabatti I, Hannant D, Paton D, Autorino GL, Nowotny N, Belák S.To determine a conclusive phylogeny, equine arteritis viruses from Italy, Austria, Hungary, Sweden, South Africa and other parts of the world were analysed by reverse-transcription polymerase chain reaction amplification and direct sequencing. The nucleotide sequences corresponding to the variable part of the large glycoprotein GP5, specified by open reading frame 5, were compared and added to a previously published phylogenetic tree in which a clear division between 'European' and 'American' type viruses had been established. Adding the sequences determined in this study and new sequences ret...
Léon A, Pronost S, Tapprest J, Foucher N, Blanchard B, André-Fontaine G, Laugier C, Fortier G, Leclercq R.Studies were carried out to determine the cause of death in a prematurely born Thoroughbred foal that died 24 hours after birth. Necropsy revealed gross lesions suggestive of septicemia. A commercial Leptospira polymerase chain reaction (PCR) assay designed to specifically amplify the hemolysis-associated protein 1 (hap1) gene present only in pathogenic Leptospira strains detected the presence of Leptospira DNA in various tissues of the foal. Histologic examination of lung, liver, kidney, and myocardium revealed numerous spirochetes in Warthin-Starry-stained tissue sections. Results of PCR ana...
Makrai L, Kira K, Kono A, Sasaki Y, Kakuda T, Tsubaki S, Fodor L, Varga J, Taka S.The plasmid profiles of virulent Rhodococcus equi strains isolated on three horse-breeding farms located in different parts of Hungary were investigated. From 49 soil samples collected on the three farms, 490 R. equi isolates (10 from each sample) were obtained and tested for the presence of 15- to 17-kDa antigens (VapA) by immunoblotting and PCR. Ninety-eight VapA-positive isolates were detected from 30 of the 49 culture-positive samples with a prevalence ranging from 13.1% to 23.2%. Of the 98 virulent isolates, 70 contained an 85-kb type I plasmid, 13 contained an 87-kb type I plasmid, and 1...
White SD, Vandenabeele SI, Drazenovich NL, Foley JE.Malassezia-type yeasts previously have been observed on cytologic examination of the intermammary region of mares that presented with tail-head pruritus; topical antiyeast treatment resolved the pruritus. Further, Malassezia dermatitis has been observed in horses in intertriginous areas such as the udder and prepuce; the species of yeast was not confirmed. It is not known whether healthy mares or male horses can be carriers of this yeast in these body areas. Objective: Malassezia spp. are present in the intermammary region in healthy mares and the preputial fossa in healthy geldings. Methods: ...
Perrocheau M, Boutreux V, Chadi S, Mata X, Decaunes P, Raudsepp T, Durkin K, Incarnato D, Iannuzzi L, Lear TL, Hirota K, Hasegawa T, Zhu B, de Jong P....A medium-density map of the horse genome (Equus caballus) was constructed using genes evenly distributed over the human genome. Three hundred and twenty-three exonic primer pairs were used to screen the INRA and the CHORI-241 equine BAC libraries by polymerase chain reaction and by filter hybridization respectively. Two hundred and thirty-seven BACs containing equine gene orthologues, confirmed by sequencing, were isolated. The BACs were localized to horse chromosomes by fluorescent in situ hybridization (FISH). Overall, 165 genes were assigned to the equine genomic map by radiation hybrid (RH...
Nugent J, Birch-Machin I, Smith KC, Mumford JA, Swann Z, Newton JR, Bowden RJ, Allen GP, Davis-Poynter N.Equid herpesvirus 1 (EHV-1) can cause a wide spectrum of diseases ranging from inapparent respiratory infection to the induction of abortion and, in extreme cases, neurological disease resulting in paralysis and ultimately death. It has been suggested that distinct strains of EHV-1 that differ in pathogenic capacity circulate in the field. In order to investigate this hypothesis, it was necessary to identify genetic markers that allow subgroups of related strains to be identified. We have determined all of the genetic differences between a neuropathogenic strain (Ab4) and a nonneuropathogenic ...
Cardoso LD, Freitas RN, Mafra CL, Neves CV, Figueira FC, Labruna MB, Gennari SM, Walker DH, Galvão MA.The present study was intended to characterize Rickettsia spp. circulating in arthropod vectors in Caratinga, Minas Gerais, Brazil, by PCR and to investigate the presence of antibodies against the spotted fever Rickettsiae group (SFRG) in dogs and horses. 2,610 arthropods were collected and taxonomically identified. DNA samples obtained from these vectors were submitted to PCR and cycle-sequenced. Ctenocephalides and Amblyomma cajennense showed sequences presenting 100.0% homology with R. felis. A sequence obtained from Rhipicephalus sanguineus showed 99.0% homology with R. felis, and a sequen...
Foote CE, Love DN, Gilkerson JR, Wellington JE, Whalley JM.A silent cycle of equine herpesvirus 1 infection was described following epidemiological studies of unvaccinated mares and foals on a Hunter Valley stud farm. Following the introduction of routine vaccination with an inactivated whole virus equine herpesvirus 1 (EHV-1) and equine herpesvirus 4 (EHV-4) vaccine in 1997, a subsequent study identified excretion of EHV-1 and EHV-4 in nasal swab samples tested by PCR from vaccinated mares and their unweaned, unvaccinated foals. The current sero-epidemiological investigation of vaccinated mares and their young foals found serological evidence of EHV-...
Pusterla N, Madigan JE, Leutenegger CM.The focus of rapid diagnosis of infectious disease of horses in the last decade has shifted from the conventional laboratory techniques of antigen detection, microscopy, and culture to molecular diagnosis of infectious agents. Equine practitioners must be able to interpret the use, limitations, and results of molecular diagnostic techniques, as they are increasingly integrated into routine microbiology laboratory protocols. Polymerase chain reaction (PCR) is the best-known and most successfully implemented diagnostic molecular technology to date. It can detect slow-growing, difficult-to-cultiv...
Scholz HC, Joseph M, Tomaso H, Al Dahouk S, Witte A, Kinne J, Hagen RM, Wernery R, Wernery U, Neubauer H.A polymerase chain reaction (PCR) assay targeting the flagellin P (fliP)-I S407A genomic region of Burkholderia mallei was developed for the specific detection of this organism in pure cultures and clinical samples from a recent outbreak of equine glanders. Primers deduced from the known fliP-IS407A sequence of B. mallei American Type Culture Collection (ATCC) 23344(T) allowed the specific amplification of a 989-bp fragment from each of the 20 B. mallei strains investigated, whereas other closely related organisms tested negative. The detection limit of the assay was 10 fg for purified DNA of ...
Hodgkinson JE.The future implementation of improved and sustainable control strategies for the major equine parasites will be dependent on a greater insight into their basic biology, pathogenicity and epidemiology together with an enhanced ability for accurate diagnosis. This paper will provide a review of the current molecular methods under development for the detection of equine parasites and their application to current scientific questions. In particular, the strongyles are recognised as important pathogens of horses and recent advances made in the study of this parasitic group at the single species lev...
Coles GC, Jackson F, Pomroy WE, Prichard RK, von Samson-Himmelstjerna G, Silvestre A, Taylor MA, Vercruysse J.Before revised World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines on the detection of anthelmintic resistance can be produced, validation of modified and new methods is required in laboratories in different parts of the world. There is a great need for improved methods of detection of anthelmintic resistance particularly for the detection of macrocyclic lactone resistance and for the detection of resistant nematodes in cattle. Therefore, revised and new methods are provided here for the detection of anthelmintic resistance in nematodes of ruminants, horses and ...
Wuersch K, Huessy D, Koch C, Oevermann A.Proliferative enteropathy (PE) caused by the obligate intracellular bacterium Lawsonia intracellularis is a disease of high economic impact in swine worldwide. In most other species the disease occurs as a sporadic infection. This paper reports a PE caused by L. intracellularis in a 9-month-old Pura Raza Española filly with a history of profuse diarrhoea. Pathological lesions consisted of a severe proliferative enteritis associated with argyrophilic bacteria in the apical cytoplasm of proliferating crypt epithelium. Characteristic PCR products confirmed the presumptive diagnosis of L. intrace...
von Samson-Himmelstjerna G.Conventional and real time polymerase chain reaction-based tests have been developed for the diagnosis of anthelmintic resistance (AR) in populations of several small and large ruminant as well as horse gastro-intestinal nematode species. To date, molecular markers that correlate well with AR are available only for the detection of benzimidazole resistance. Recently, however, a single nucleotide polymorphism was found in vitro to be of functional relevance for reduced drug efficacy to macrocylic lactones. The focus of the present review, therefore, is the molecular mechanism of action of these...
Al-Ghamdi GM, Kapur V, Ames TR, Timoney JF, Love DN, Mellencamp MA.To determine whether repetitive sequence-based polymerase chain reaction (rep-PCR) could be used to differentiate Streptococcus equi isolates, to examine S equi isolates from throughout the world, and to determine whether a horse had > 1 subtype of S equi during an outbreak of disease. Methods: An initial group of 32 S equi isolates, 63 S equi isolates from various geographic areas, and 17 S equi isolates obtained during outbreaks of disease. Methods: An aliquot of S equi genomic DNA was amplified, using enterobacterial repetitive intergenic consensus primers. Gel electrophoresis was perfor...
Takai S, Zhuang D, Huo XW, Madarame H, Gao MH, Tan ZT, Gao SC, Yan LJ, Guo CM, Zhou XF, Hatori F, Sasaki Y, Kakuda T, Tsubaki S.Little is known about the distribution of Rhodococcus equi in the soil environment of native horses in China. One hundred and eight soil samples were collected from native-horse farms in the Hulun Beier grasslands of eastern Mongolia, the Xilin Goler grasslands of southern Mongolia, and Tongliao City in Inner Mongolia, China. The isolation rates of R. equi from soil samples from the Hulun Beier and Xilin Goler grasslands ranged from 25.9% to 30.0%. In contrast, isolation rates from soil samples from Tongliao City were as high as 82.3% and the mean number of R. equi in soil samples from Tonglia...
Zarski LM, High EA, Nelli RK, Bolin SR, Williams KJ, Hussey G.Equine herpesvirus 5 (EHV-5) infection is associated with pulmonary fibrosis in horses, but further studies on EHV-5 persistence in equine cells are needed to fully understand viral and host contributions to disease pathogenesis. Our aim was to develop a quantitative PCR (qPCR) assay to measure EHV-5 viral copy number in equine cell cultures, blood lymphocytes, and nasal swabs of horses. Furthermore, we used a recently developed equine primary respiratory cell culture system to study EHV-5 pathogenesis at the respiratory tract. PCR primers and a probe were designed to target gene E11 of the EH...
Albert R, Vásárhelyi G, Bodó G, Kenyeres A, Wolf E, Papp T, Terdik T, Módis L, Felszeghy S.One of the most promising applications for the restoration of small or moderately sized focal articular lesions is mosaicplasty (MP). Although recurrent hemarthrosis is a rare complication after MP, recently, various strategies have been designed to find an effective filling material to prevent postoperative bleeding from the donor site. The porous biodegradable polymer Polyactive (PA; a polyethylene glycol terephthalate - polybutylene terephthalate copolymer) represents a promising solution in this respect. A histological evaluation of the longterm PA-filled donor sites obtained from 10 exper...
Velineni S, Timoney JF.Streptococcus zooepidemicus causes opportunist respiratory and other infections in the horse. Capsule expression is highly variable and known to affect resistance to phagocytosis. Most clinical isolates producing small, dry colonies at 37°C produce mucoid colonies at temperatures below 35°C. Objective: The aim was to understand the molecular basis of increased capsule expression by equine isolates of S. zooepidemicus at temperatures lower than 35°C. Methods: Cross-sectional observational study. Methods: Capsule production by groups of equine S. zooepidemicus strains was determined at 23, ...
Eggleston-Stott ML, Delvalle A, Dileanis S, Wictum E, Bowling AT.The equine dinucleotide microsatellite HMS7 is part of a microsatellite panel utilized in a parentage verification programme at the Veterinary Genetics Laboratory (Davis, California, USA). Apparent non-Mendelian inheritance was noted when a Quarter Horse mare was excluded as the parent of two offspring based on analysis of the HMS7 locus. The mare's DNA type qualified her as a parent of the offspring at an additional 20 microsatellite loci. The three animals appeared homozygous for HMS7 with each possessing an allele different from that of the other two animals. Polymerase chain reaction prime...
Annandale A, Stroehle RM, Schulman ML, Sibeko-Matjila KP, Fosgate GT, Handler J, Vemming DC, Clift SJ.Persistent breeding-induced endometritis (PBIE) or delayed uterine clearance (DUC) are major causes of mare subfertility. Oxytocin and its receptor are thought to play significant roles in the pathogenesis of DUC but the specific roles of oxytocin receptor (OR) distribution and gene expression remain undefined. In this study both OR distribution and gene expression in the endometrium, myometrium and cervix during both luteal and non-luteal phases in non-pregnant mares (n = 27) of differing age (young: 2-9 years, n = 17; old: > 10 years, n = 10) and endometrial biopsy score were ...
Schöniger S, Gräfe H, Wipplinger M, Schoon HA.In mares, placental diseases are a common cause of pregnancy failure and they can have an economic impact on the horse breeding industry. To our knowledge no published data on TLR expression in the equine placenta exist. This study examined the expression of TLR 2, 4 and 6 as transcript and protein in the placenta (chorioallantois) of 14 foals born alive. By PCR, all examined placental samples contained TLR 2, 4 and 6 transcripts. Using immunohistochemistry, trophoblasts and allantoic epithelium were immunopositive for TLR 2, 4 and 6 in all placental samples. The majority of placental samples ...
Davidson AJ, Edwards GB, Proudman CJ, Cripps PJ, Matthews JB.The aim of this study was to investigate cytokine expression patterns in the large intestinal mucosa of horses, particularly in diseases associated with inflammation. Many cases of equine colitis remain without a definitive diagnosis and survival rates are poor. In humans, colitis is associated with increased expression of pro-inflammatory cytokines. To examine if similar responses occur in horses, we investigated il -2, il -4, il -5, il -10, tnfalpha, ifngamma and tgfbeta messenger rna expression in large intestinal mucosa. Samples were obtained from animals with large intestinal disease (n=1...
Narváez SÁ, Fernández I, Patel NV, Sánchez S. is an important veterinary pathogen that takes the lives of many foals every year. With the emergence and spread of MDR to current antimicrobial treatment, new tools that can provide a fast and accurate diagnosis of the disease and antimicrobial resistance profile are needed. Here, we have developed and analytically validated a multiplex qPCR for the simultaneous detection of and related macrolide resistance genes in equine respiratory samples. The three sets of oligos designed in this study to identify housekeeping gene and macrolide resistance genes (46) and (51) showed high analytic se...
Hollingsworth SR, Pusterla N, Kass PH, Good KL, Brault SA, Maggs DJ.To determine the role of equine herpesvirus (EHV) in idiopathic keratoconjunctivitis in horses and to determine whether sample collection method affects detection of EHV DNA by quantitative polymerase chain reaction (qPCR). Methods: Twelve horses with idiopathic keratoconjunctivitis and six horses without signs of ophthalmic disease. Methods: Conjunctival swabs, corneal scrapings, and conjunctival biopsies were collected from 18 horses: 12 clinical cases with idiopathic keratoconjunctivitis and six euthanized controls. In horses with both eyes involved, the samples were taken from the eye judg...
Baczynska A, Fedder J, Schougaard H, Christiansen G.The reproduction rate of horses is one of the lowest within domestic livestock despite advances the veterinary medicine. Infertility in horses may be due mainly to the lack of suitable selection criteria in the breeding of horses. However, acquired infertility due to genital, bacterial infections may occur. Mycoplasmas have been implicated in genital disorders and infertility of many species including humans and horses. However, their role as commensals or pathogens of the genital tract of horses is still not determined. Bacteriological examinations made on the fossa glandis, urethra, penis an...
Imhasly A, Frey CF, Mathis A, Straub R, Gerber V.The protozoon parasite Cryptosporidium parvum is an important cause of diarrhea in farm animals, but it can also infect other animals and humans. In this case report, oocysts of Cryptosporidium spp. were microscopically detected by modified Ziehl-Neelsen staining in the feces of a 9 day old Arabian colt presented with yellowish, foul smelling, diarrhea and fever of 40 degrees C. PCR and sequencing of the isolate revealed C. parvum (bovine genotype). Hemato-chemical analysis of the foals blood revealed a marked hypogammaglobulinaemia (IgG 108mg/dl). The colt responded well to a supportive thera...
Vallone LV, Neaderland MH, Ledbetter EC, Dubielzig RR.To describe clinical, in vivo confocal microscopic, histopathologic, polymerase chain reaction assay for antigen receptor rearrangement (PARR) and immunohistochemical features of a horse with suspected malignant transformation of corneal B lymphocytes in immune-mediated keratitis (IMMK) and to also report surgical management and adjunctive treatment including topical and radiation therapies. Methods: A 15-year-old Oldenburg gelding was examined for presumptive progressive stromal IMMK of the right eye which became refractory to medical management. Corneal in vivo confocal microscopy revealed...
Ramina A, Dalla Valle L, De Mas S, Tisato E, Zuin A, Renier M, Cuteri V, Valente C, Cancellotti FM.The reverse transcriptase polymerase chain reaction (RT-PCR) assay was used to detect Equine Arteritis Virus (EAV) in the semen of 88 horses and 2 donkeys, with neutralising antibodies against EAV, on the basis of the amplification of a 279 bp long fragment located in the viral polymerase gene. The RT-PCR assay revealed the virus at 4 TCID50/ml in cell culture and showed a greater sensitivity (54.4%) than cell culture isolation (33.3%). Moreover, the two samples of donkey semen were found positive. The cDNAs obtained from 14 samples of horse and 2 of donkey semen were sequenced. Comparing the ...
Kalantari M, Sharifiyazdi H, Ghaemi M, Ghane M, Nazifi S.In all equids worldwide, Theileria equi and Babesia caballi are believed to be two important erythrocytic protozoa that cause equine piroplasmosis. In addition, it was recently discovered that Theileria haneyi is another potential equine piroplasmosis (EP) agent. Ixodid ticks are the major vectors of these parasites. Equine piroplasmosis is of international importance and affects enormously the equine industry. In this study, for the first time, molecular prevalence and genetic diversity of piroplasma parasites (T. equi and B. caballi) in horses from Fars province (south of Iran) were determin...
Stasiak K, Dunowska M, Rola J.Equid herpesvirus 2 (EHV-2) and 5 (EHV-5) are two γ-herpesviruses that are commonly detected from horses worldwide, based on several cross-sectional molecular surveys. Comparatively few studies examined the dynamics of γ-herpesvirus infection over time in a group of horses. The aim of the current study was to investigate the dynamics of EHV-2/5 infections among mares and their foals at three Polish national studs with different breeds of horses: Arabians, Thoroughbreds and Polish Konik horses. Nasal swabs were collected from each of 38 mare-foal pairs monthly for a period of 6 to 8 months. V...
Ataseven VS, Oğuzoğlu TÇ, Başaran-Karapınar Z, Bilge-Dağalp S.Equine adenovirus type 1 (EAdV-1) is a cause of repiratory tract infection in equids. In present study for the first time in Turkey, the prevalence of EAdV-1 in nasal swab samples obtained from horses showing respiratory symptoms was investigated by polymerase chain reaction (PCR), and molecular characterization of the hexon gene detected in the Turkish (TR) strain was performed. Overall, the prevalence of EAdV-1 was found low (1.4%) as indicated by a positive PCR reaction from the nasal swab extracts tested. Phylogenetic analysis based on the partial sequences of the hexon gene of a TR-EAdV-1...
Zakia LS, Olivo G, Basso RM, Mira J, Herman M, Araujo JP, Borges AS, Oliveira-Filho JP.Aural plaques are a dermatopathy associated with Equus caballus papillomavirus (EcPV). This disease affects horses of all ages, genders and breeds, and causes sensitivity of the ears. Objective: The aim of this study was to evaluate the clinical efficacy of 5% imiquimod cream for the treatment of aural plaques and to compare the PCR detection of EcPV 3, 4, 5 and 6 before and after treatment. Methods: Eight horses diagnosed with aural plaques (14 ears) were used. Three mares with unilateral aural plaques were used as untreated controls. Methods: Imiquimod cream was applied every 48 h until comp...
King SS, Jones KL, Mullenix BA, Heath DT.Dopamine (DA) blockade during anestrus or early spring transition can facilitate ovarian recrudescence and advance the timing of the first ovulation of the season. Some laboratories have reported variable results using DA antagonists to stimulate follicular growth during the mid-portion of the anestrual period. Differences in DA antagonist efficacy may be due to the FSH secretory status of the anestrous mare and the presence or absence of functional ovarian FSH receptors. We hypothesize that direct ovarian dopaminergic input can affect follicular growth through regulation of FSH receptor (FSHr...
El-Gameel SM, Al-Mokaddem AK, Salaeh NMK, Attia MM.Equine gastrointestinal tract is infected with () which is highly pathogenic parasite for its harmful effect on cranial mesenteric artery during its migration. So, this study was applied for identification of in donkeys ultramorphologically and molecularly. In addition to, detection of the pathological effect of larval stage of on the mesenteric arterial system using histopathology and immunohistochemistry. During the period from September to December; 2019, 60 male and 20 female donkeys at the Giza Zoo was postmortem examined. adults and larvae were collected from the large intestine and ...
Shin EK, Perryman LE, Meek K.To determine whether a recently developed test would correctly identify horses heterozygous for the severe combined immunodeficiency (SCID) trait. Methods: Case series. Methods: 17 healthy Arabian horses that had previously produced foals with SCID, 1 healthy Arabian foal whose dam and sire had produced foals with SCID, 4 foals with SCID, and 1 healthy non-Arabian foal. Methods: DNA was extracted from leukocytes or fibroblasts, amplified by means of polymerase chain reaction, and hybridized with probes specific for the normal and mutant alleles of the catalytic subunit of DNA-dependent protein...
Miglinci L, Reicher P, Nell B, Koch M, Jindra C, Brandt S.Squamous cell carcinoma (SCC) seriously compromises the health and welfare of affected horses. Although robust evidence points to equine papillomavirus type 2 (EcPV2) causing genital lesions, the etiopathogenesis of equine SCC is still poorly understood. We screened a series of SCCs from the head-and-neck (HN), (peri-)ocular and genital region, and site-matched controls for the presence of EcPV2-5 and herpesvirus DNA using type-specific EcPV PCR, and consensus nested herpesvirus PCR followed by sequencing. EcPV2 DNA was detected in 45.5% of HN lesions, 8.3% of (peri-)ocular SCCs, and 100% of g...
Bogaert L, Heerden MV, Cock HE, Martens A, Chiers K.Ten equine skin tumors that had been classified as schwannomas on routine histological examination were analyzed by polymerase chain reaction for bovine papillomavirus DNA. All 10 were positive for bovine papillomavirus 1 or 2, and all 10 were immunohistochemically negative for S-100 protein and strongly positive for vimentin. Nine tumors were moderately positive for laminin and 8, for smooth muscle actin. Five tumors were variably and weakly positive for type IV collagen. The lack of S-100 protein expression made Schwann cells an unlikely cell of origin, as opposed to peripheral nerve sheath ...
Kim SH, Lee JS, Lee JH, Kim YJ, Choi JG, Lee SK, Kim HJ, Yang SJ, Park T, Lee SK, Kang HE, Jeoung HY, Park JY.Venereal diseases caused by bacteria are important to the equine industry due to economic losses caused by decline of conception rate in breeding horses. Therefore, identification of infected animals as well as the implementation of appropriate managerial procedures based on accurate diagnosis is critical. In this study, two types of multiplex real-time polymerase chain reaction with high sensitivity and specificity were developed for the simultaneous detection and differentiation of five commonly associated bacterial pathogens of venereal diseases in horses, consisting of Taylorella equigenit...
Papapetrou MA, Arroyo LG, Meister TL, Baird JD, Steinmann E, Lillie BN.Equine parvovirus-hepatitis (EqPV-H) was first reported from the serum and liver tissue of a horse diagnosed with Theiler's disease in the United States in 2018. Theiler's disease, also known as equine serum hepatitis, is a severe hepatitis with fulminant hepatic necrosis. The disease has most frequently been reported following the administration of equine-origin biological products; however, it has also been reported in in-contact horses with no prior biologic administration. EqPV-H has been detected in clinically healthy horses in North America (USA, Canada), Europe (Germany, Austria, Sloven...
Romo-Sáenz CI, Tamez-Guerra P, Olivas-Holguin A, Ramos-Zayas Y, Obregón-Macías N, González-Ochoa G, Zavala-Díaz de la Serna FJ....Equine infectious anemia (EIA) is a highly infectious disease in members of the 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 immu...
Zahoor J, Kashif M, Nasir A, Bakhsh M, Qamar MF, Sikandar A, Rehman A.Trypanosomiasis is one of the severe pathogenic infections, caused by several Trypanosoma species, affecting both animals and humans, causing substantial economic losses and severe illness. The objective of this study was to determine the molecular diagnosis and the risk factors associated with trypanosomiasis in District Jhang, Punjab, Pakistan. For this purpose, blood samples were randomly collected from 200 horses. A predesigned questionnaire was used to collect data on risk factors before the sample collection. The microscopy examination through Giemsa staining, formol gel test and PCR tec...
Coffman EA, Abd-Eldaim M, Craig LE.A pregnant 18-year-old Quarterhorse mare presented with fever, anorexia, tachycardia, tachypnea, and gastrointestinal hypermotility at day 68 of gestation. Potomac horse fever was diagnosed based on polymerase chain reaction (PCR) analysis of whole blood and a high antibody titer to Neorickettsia risticii. The mare made a rapid clinical recovery following antibiotic therapy, but aborted 98 days later. Necropsy on the aborted fetus revealed lymphohistiocytic colitis, lymphadenitis, myocarditis, and hepatitis. The placenta was grossly and histologically normal. Formalin-fixed lymph node, thymus,...
Pusterla N, Barnum SM, Byrne BA.Polymerase chain reaction (PCR)-based detection assays for Streptococcus equi subspecies equi often overestimate the prevalence of samples containing viable organisms. The objective of this study was to determine if viability could be determined using genome quantitation and detection of messenger RNA (mRNA) transcripts for the SeM gene of S. equi in pre- and post-cultured samples. Nasal secretions collected from 42 horses with suspected strangles were tested by culture and by quantitative PCR (qPCR) before and 24 hours after a culture step. Viable S. equi was determined based on the detecti...
