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.
Gilkerson J, Jorm LR, Love DN, Lawrence GL, Whalley JM.Equid herpesvirus-4 (EHV-4) was detected in nasal swabs taken from foals using a PCR based test and this information used to study the epidemiology of EHV-4 disease on three Australian Thoroughbred stud farms in NSW in 1992. There was a very high level of agreement (kappa value of 0.84) between the PCR results and virus isolation using cell culture techniques. There was a strong seasonal distribution of EHV-4 shedding. Twenty-five of 26 positive samples were collected in January and March with the remaining positive sample collected in February. Foals with clinical signs of upper respiratory t...
Kim CH, Casey JW.The distribution and replicative status of equine infectious anemia virus (EIAV) DNA in the tissues of a well-characterized inapparent carrier horse were established by using the PCR technique. The EIAV pol region could be amplified in all of the tissues tested, including the cerebellum and periventricular tissue, at concentrations approximately 10(5)-fold less than in the same tissue from an acutely infected horse. Further analysis of the EIAV genome, with primer pairs diagnostic for sequential stages of reverse transcription, suggests that EIAV DNA in the brain, liver, and lymph nodes was in...
Bleumink-Pluym NM, Werdler ME, Houwers DJ, Parlevliet JM, Colenbrander B, van der Zeijst BA.A PCR for the detection of Taylorella equigenitalis, the causative agent of contagious equine metritis, was developed and evaluated. A genus-specific primer-probe set was derived from the 16S ribosomal DNA sequences. The PCR was specific and amplified a 585-bp product from all 64 available T. equigenitalis isolates. This PCR product hybridized with a specific probe in a dot spot assay. A variety of microorganisms from the genital tracts of horses or with a close phylogenetic relationship to T. equigenitalis did not yield a visible PCR product and were all negative in the dot spot hybridization...
Lawrence GL, Gilkerson J, Love DN, Sabine M, Whalley JM.Sets of primers were designed which enabled specific amplification of homologous regions of the glycoprotein C and gene 76 genetic loci of equine herpesviruses 1 and 4 (EHV-1 and EHV-4). The resultant virus-specific polymerase chain reaction (PCR) products arising from each loci could be discriminated easily on the basis of size on an agarose gel, allowing rapid differentiation of the two equine herpesviruses. Specificity of the amplifications were confirmed by Southern hybridization and restriction endonuclease digestion. The PCR test was applied to nasal swab samples from weanling foals and ...
Wöhrl BM, Howard KJ, Jacques PS, Le Grice SF.A comparative study of recombinant 51- and 66-kDa subunits comprising equine infectious anemia virus reverse transcriptase (EIAV RT) is reported. Both polypeptides sedimented as stable homodimers (molecular mass, 102 and 132 kDa, respectively) when analyzed by rate sedimentation through glycerol gradients. Consistent with their dimer composition, each preparation displayed considerable levels of both RNA- and DNA-dependent DNA polymerase activity on different homopolymeric template/primer combinations. However, a detailed analysis of the polymerization products indicated qualitative difference...
Edington N, Welch HM, Griffiths L.Equid herpesviruses 1 or 4 (EHV-1 or -4) were isolated by cocultivation from 60% of 40 horses examined at slaughter. The lymph nodes draining the respiratory tract were the most common source of virus. EHV-1 or EHV-4 was never isolated from the trigeminal ganglia (SLG). The polymerase chain reaction (PCR) detected virus in 87.5% of bronchial lymph nodes and a similar level in the trigeminal ganglia that were examined. By both assays approximately one third of the positive animals harboured both viruses. Equid herpesvirus 2 (EHV-2) was isolated from all but one of the horses and from > 75% o...
St-Laurent G, Morin G, Archambault D.A reverse transcription (RT)-PCR assay was developed for the detection of equine arteritis virus (EAV) in cell culture supernatant and in horse semen. Four different sets of oligonucleotide primers complementary to sequences located in the 3' end of the polymerase gene (open reading frame [ORF] 1b) and to sequences representing the entire ORFs 3, 4, and 7, which encode for nonstructural (ORFs 3 and 4) or viral nucleocapsid (ORF 7) proteins, were compared for their abilities to amplify the targeted EAV sequences by the RT-PCR procedure. The sensitivities of the RT-PCR for amplification of EAV s...
Stone-Marschat M, Carville A, Skowronek A, Laegreid WW.Reverse transcription-PCR (RT-PCR) was used to detect African horse sickness virus (AHSV). A single primer pair which amplified a 423-bp fragment of the S8 gene which encodes the NS2 protein of AHSV was identified. Amplification of this fragment from all nine serotypes of AHSV was achieved with these primers. Between 10(1) and 10(2) copies of AHSV genomic double-stranded RNA could be detected by RT-PCR followed by agarose gel electrophoresis and ethidium bromide staining. Application of RT-PCR to blood samples from AHSV-infected horses resulted in earlier detection of viremia than virus isolat...
Zimmermann W, Dürrwald R, Ludwig H.Borna disease virus in naturally infected horses, a donkey and sheep was detected for the first time by amplification of viral RNA using PCR. In contrast to a control group of healthy horses, brain tissue was positive by this assay in all animals with neurological symptoms. The use of a second round of PCR with nested primers following Southern hybridization confirmed the specificity and increased the sensitivity of the test. Comparison with conventional methods recommends this technique for monitoring of BDV infections at a molecular level.
Pilling A, Davison AJ, Telford EA, Meredith DM.Glycoprotein 45 is a major envelope glycoprotein of equine herpesvirus type 1. The gene encoding this protein is located between map units 0.615 and 0.636 on the virus genome and evidence has suggested that it is encoded by gene 52, one of four genes within this region. Using PCR we have amplified gene 52 and subsequently cloned it into a mammalian expression vector under the control of the human cytomegalovirus immediate early gene promoter. The gene was expressed in COS-7 cells and its product was detected by immunofluorescence and Western blotting. The results indicate that glycoprotein 45 ...
Marklund S, Ellegren H, Eriksson S, Sandberg K, Andersson L.Ten (TG)n positive clones, isolated from an equine genomic library and sequenced, contained 12-19 uninterrupted TG repeats. Primers for polymerase chain reaction (PCR) were synthesized and nine of these (TG)n loci (HTG7-15) were successfully amplified and utilized in this study together with five previously reported equine microsatellite loci (HTG2-6). The PCR products were analysed by polyacrylamide gel electrophoresis followed by automated laser fluorescence detection or autoradiography. All microsatellites showed polymorphism and stable Mendelian inheritance. Differences in microsatellite v...
Zientara S, Sailleau C, Moulay S, Cruciere C.A single tube reverse transcription-polymerase chain reaction (RT-PCR) method for detection of African horse sickness virus (AHSV) in splenic tissues from infected horses is described. Double stranded RNA was extracted from infected organs of horses and used to produce complementary DNA (cDNA) with the two primers selected for the PCR. The 1179 bp amplified product (the segment 7 which encodes for VP 7), detected by electrophoresis on agarose gel and ethidium bromide staining, was hydrolysed with eight restriction endonucleases for characterization of the AHSV. The sensitivity of this method i...
Donofrio JC, Coonrod JD, Chambers TM.Influenza A is a common respiratory infection of horses, and rapid diagnosis is important for its detection and control. Sensitive detection of influenza currently requires viral culture and is not always feasible. The polymerase chain reaction (PCR) was used to detect DNA produced by reverse transcription of equine influenza in stored nasal secretions, vaccines, and allantoic fluids. Primers directed at a target of 212 bp on conserved segment 7 (matrix gene) of human influenza A/Bangkok/1/79(H3N2) produced amplification products of appropriate size with influenza A/Equine/Prague/1/56 (H7N7), ...
Vandergrifft EV, Horohov DW.We have cloned equine IL-2 cDNA in vitro using the polymerase chain reaction (PCR) and primers based on the human IL-2 sequence. The cloned product appears to contain the entire coding region for equine IL-2 based on homology with other known sequences. When expressed in COS cells, the recombinant product augmented the proliferative response of equine peripheral blood mononuclear cells to concanavalin A, however, it failed to support the continued proliferation of murine CTLL-2 cells. Specific substitutions in those regions associated with p55 and p75 binding appear to account for this species...
Vodkin MH, McLaughlin GL, Day JF, Shope RE, Novak RJ.Eastern equine encephalomyelitis virus (EEEV) has been a low-frequency, but serious human and veterinary health problem. Increased frequency of this mosquito-borne virus is anticipated as wetlands are maintained and re-established. Control of EEEV has depended on mosquito abatement in response to increasing frequency of EEEV in the environment. A coupled reverse transcription/polymerase chain reaction assay was designed to rapidly, sensitively, and specifically detect EEEV RNA. The assay successfully detected the viral RNA in a single-blind study of a set of field samples composed of either po...
Palmer JE.E. risticii, the cause of classic Potomac horse fever, is now known to produce two disease syndromes: EEC and EEA. The pathogen appears to commonly infect horses based on seroepidemiologic studies; however, the method of transmission remains unknown. The most common clinical disease is EEC, commonly called Potomac horse fever, which presents a wide spectrum of clinical signs. Diagnosis is currently dependent on serology, which frequently does not lead to a definitive diagnosis and at best results in a retrospective diagnosis. A new diagnostic approach, polymerase chain reaction, may offer a ra...
Greeve J, Altkemper I, Dieterich JH, Greten H, Windler E.Two different isoproteins are encoded by the apolipoprotein (apo) B gene, apoB-48 and apoB-100. ApoB-48, core component of intestinally derived chylomicrons, has an accelerated plasma turnover as compared with the full-length protein apoB-100. A posttranscriptional modification of the apoB mRNA by conversion of cytidine into uridine at nucleotide position 6666 changes the genomically encoded glutamine codon CAA at amino acid residue 2153 into a translational stop codon UAA. This mRNA editing explains the formation of the truncated isoform apoB-48. In the present investigation editing of apoB m...
Räsänen LA, Karvonen U, Pösö AR.In situ hybridization was used to localize xanthine dehydrogenase (XDH) mRNA in horse skeletal muscle. Capillary endothelial cells were found to express XDH, but muscle cells did not give any signal. The digoxigenin-labelled probe was produced by PCR with primers based on the cDNA sequence of mouse XDH and horse lung cDNAs. A 4.3 kb mRNA was detected in a Northern blot.
Szalai G, Bailey E, Gerber H, Lazary S.The genetic diversity at the ELA DQ beta locus was investigated using polymerase chain reaction and DNA sequencing. Based upon serological methods 16 class II homozygous animals were selected and their genomic DNA was used. A DQ beta gene from an equine cDNA library was also sequenced. Our methodology and the similarity between the genomic and the cDNA sequences suggest that the studied locus is expressed on equine lymphocytes. In the predicted amino acid sequence the most extensive variation is located at residues 56-60. The pattern of these five amino acids is strongly correlated to the sero...
Rimstad E, Evensen O.Paraffin-embedded organ samples from 28 aborted fetuses and three foals, partly archival and partly sampled in 1991, were examined by polymerase chain reaction (PCR) and immunohistochemistry for the presence of DNA and antigens, respectively, specific for equine herpesvirus 1 (EHV-1). Virologic examination had been performed on 23 of the aborted fetuses. DNA fragments specific for EHV-1 were identified by PCR, and EHV-1 antigens were identified in situ by immunohistochemistry, with an agreement between the methods of 94% (kappa = 0.85). Compared with virus isolation, PCR agreement was 87% (kap...
Beisel CE, Edwards JF, Dunn LL, Rice NR.Transcription of pathogenic equine infectious anemia virus (EIAV) in an acutely infected horse was examined by using the polymerase chain reaction and nucleotide sequencing. Four spliced transcripts were identified in liver tissue, in contrast to the multiplicity of alternatively spliced messages reported for in vitro-propagated human immunodeficiency virus, simian immunodeficiency virus, and, to a lesser extent, EIAV. Nucleotide sequence analysis demonstrated that three of these mRNAs encode known viral proteins: the envelope precursor, the product of the S2 open reading frame, and the regula...
Otten N, von Tscharner C, Lazary S, Antczak DF, Gerber H.Nucleotide sequences of bovine papillomavirus (BPV) DNA amplified by the polymerase chain reaction (PCR) from samples of equine sarcoid skin tumours were determined. All naturally occurring sarcoids (n = 58 tumours from 32 horses and 2 donkeys) contained BPV-DNA. All but 3 of the genome fragments belonged to the BPV type 1 strain (BPV-1); the remaining were BPV type 2. Similar results were obtained with cutaneous bovine papillomas used as controls (n = 20). One of the horses, carrying 2 sarcoids, was particularly interesting; one tumour contained BPV-1 DNA whilst the other sarcoid yielded BPV-...
Binns MM, Daly JM, Chirnside ED, Mumford JA, Wood JM, Richards CM, Daniels RS.The haemagglutinin (HA) gene from the equine influenza H3N8 isolate Suffolk/89 has been cloned by reverse transcription and polymerase chain reaction amplification. The nucleotide sequence of the HA gene was determined from two independently cloned copies of the gene and was found to be most closely related to recent American isolates supporting the idea that most isolates of equine H3N8 are evolving as a single lineage. When the predicted amino acid sequence of the Suffolk/89 HA was examined, changes had taken place in at least four of the major antigenic sites, A, B, C, and D when compared t...
Smith KC, Whitwell KE, Binns MM, Dolby CA, Hannant D, Mumford JA.From 1988 to 1991, 51 pregnant pony mares were challenged intranasally or by aerosol with an isolate of EHV-1 (AB4) originally recovered from a quadriplegic mare. This resulted in 32 abortions, occurring from 9 to 29 days after infection. In 14 of the early abortions (Days 9-14), EHV-1 was not demonstrated in the foetal tissues by virus isolation or immunostaining despite no other non-viral cause for the abortion being evident. Application of the polymerase chain reaction to foetal tissues from 9 of these cases also proved negative. One of the 14 mares was destroyed immediately after abortion,...
Kim CH, Casey JW.Equine infectious anemia virus (EIAV) is a lentivirus that infects and persists in the monocyte/macrophage populations of blood and tissues. We employed polymerase chain reaction to investigate the distribution and the level of genome variability of EIAV DNA in different tissues of a horse infected with a highly virulent variant of the Wyoming strain of the virus. Long terminal repeat, gag, and pol primer pairs were used to direct the amplification of EIAV DNA from the peripheral blood mononuclear cells and from cells, presumably the macrophage subtypes, of the kidney, spleen, liver, lymph nod...
Sherman GB, Wolfe MW, Farmerie TA, Clay CM, Threadgill DS, Sharp DC, Nilson JH.Equine (e) CG and LH beta-subunits have identical amino acid sequences, including an extended carboxyl-terminal peptide (CTP). This suggests that unlike the corresponding human genes, the beta-subunits of eCG and eLH may be encoded by a single gene and share a common proximal promotor region. To explore this, we isolated and characterized the eLH/CG beta gene(s). Data from Southern analyses suggest that the eCG beta and eLH beta subunits are products of the same single copy gene (eLH/CG beta). Overlapping fragments of the eLH/CG beta gene and cDNA were amplified from equine genomic DNA and pit...
Welch HM, Bridges CG, Lyon AM, Griffiths L, Edington N.The polymerase chain reaction (PCR) and co-cultivation were used to identify the lymphoreticular system as the site of latency of equid herpesvirus I (EHV-1). Primers for PCR were designed from aligned nucleotide sequences of the glycoprotein gB genes to amplify the same region of both the EHV-1 and EHV-4 genomes. Subsequent restriction digests using specific enzymes distinguished the amplified fragments of the EHV-1 genome from those of the EHV-4 genome. Ten weeks following an experimental infection of five ponies with EHV-1, latent virus was detected by PCR and recovered by co-cultivation, p...
Hardt M, Teifke JP, Weiss E.Formalin-fixed and Paraplast-embedded tissue samples of 42 aborted equine fetuses were examined by polymerase chain reaction for the presence of equine herpesvirus DNA. The used set of primers was located in the glycoprotein 13 open reading frame and allowed the amplification of both EHV 1 und EHV 4. By cleaving pattern analysis after Hinf I digestion EHV 1 could be distinguished from EHV 4. In 9 of the cases investigated EHV 1-DNA was detected. This finding is in absolute context with the results of the virological investigations.
Fernandes TA, Paulino PG, Dos Santos Juliano D, Rabello CA, de Oliveira NVB, de Souza Santana M, Peckle M, Massard CL, da Costa Angelo I, Jacob JCF....Equine piroplasmosis (EP) is a reportable disease according to the World Organization for Animal Health (WOAH), caused by Babesia caballi, Theileria equi and T. haneyi. This syndrome is prevalent in tropical and subtropical regions, including various areas in Brazilian. This study examines EP in the Distrito Federal, Brazil, focusing on prevalence, epidemiological characteristics, and circulating genotypes. Epidemiological data and whole blood samples were collected from horses in the Distrito Federal. DNA was analyzed using qPCR for Theileria sp. and B. caballi, followed by cPCR for 18S rDNA ...
Sun Y, Yu YT, Castillo XO, Anderson R, Wang M, Sun Q, Tallmadge R, Sams K, Reboul G, Zehr J, Brown J, Wang X, Marra N, Stanhope B, Grenier J....Fever of unknown origin (FUO) without a respiratory component is a frequent clinical presentation in horses. Multiple pathogens, both tick-borne and enteric, can be involved as etiologic agents. An additional potential mechanism is intestinal barrier dysfunction. This case-control study aimed to detect and associate microbial taxa in blood with disease state. Areas known for a high prevalence of tick-borne diseases in humans were chosen to survey horses with FUO, which was defined as fever of 101.5°F or higher with no signs of respiratory illness or other recognisable diseases. Blood samples ...
Kawanishi N, Kinoshita Y, Reedy SE, Garvey M, Kambayashi Y, Bannai H, Tsujimura K, Yamanaka T, Cullinane A, Chambers TM, Nemoto M.Equine influenza virus (EIV) is one of the most important pathogens causing respiratory signs in equids. Rapid antigen detection (RAD) kits are useful for point-of-care testing because they are user-friendly and provide fast results. Although sensitive and broad-reactive RAD kits are needed for controlling horse movement, no RAD kits specified for EIV are available. Objective: This study evaluated the usefulness of seven RAD kits originally developed for human influenza and available in Japan during 2023-2024 for EIV antigen detection. Methods: Experimental assay comparison. Methods: The detec...
Peris MP, Serrano M, Romero A, García M, Halaihel N, Castillo JA, Gracia MJ.Equine piroplasmosis (EP), a tick-borne disease caused by Babesia caballi and Theileria equi, is of significant concern due to its impact on the international horse trade. According to standards established by the World Organisation for Animal Health (WOAH), horses imported from EP-endemic regions must have a certificate confirming negative serological and molecular test results. In EP-free countries, only seronegative horses are allowed to enter. This entails economic losses for endemic regions such as Spain. Our study aimed to assess the prevalence of B. caballi and T. equi in horses from no...
Pusterla N, Lawton K, Barnum S, Magdesian KG.In recent years, the use of non-invasive host and environmental samples for the detection and monitoring of equine respiratory pathogens has shown promise and a high overall agreement with the gold standard of nasal secretions. The present study looked at comparing nose wipes, stall sponges, and air samples with nasal swabs collected from 27 horses involved in an equine influenza (EI) outbreak. The outbreak involved 5 clinical, 6 subclinical, and 16 uninfected horses. Samples sets were collected at the onset of the index case and retested every 2-3 days thereafter until all horses tested qPCR-...
Polo MC, Huby FD, Uehlinger FD, Rubin JE.Antimicrobial resistance is a serious threat in human and veterinary medicine. Among the most problematic resistant organisms are the extended-spectrum β-lactamase (ESBL)-producing Enterobacterales, which are resistant to the 3rd-generation cephalosporins. The purpose of this study was to determine the frequency of colonization of horses admitted to the Western College of Veterinary Medicine with resistant Escherichia coli. Unassigned: Rectal swabs were collected from 60 horses admitted between November 2021 and March 2022. Swabs were selectively cultured for E. coli, which was identified usi...
Hosseinzadeh S, Nofouzi K, Hasanzadeh F, Esmaeili S, Ayen E.Q fever is a frequently occurring illness that is induced by the bacterium ) that can infect humans and various animals. It targets the macrophage cells in the tissues, and circulating monocytes. Unassigned: This study was conducted between 2022 and 2023 in the West Azerbaijan and Ardabil provinces of northwestern Iran to examine the presence infection of . Specimens were obtained by swabbing from 140 mares (70 from each province) and 20 stallions (10 from each province) which were apparently healthy, and their DNA was analyzed using quantitative PCR assay detecting the element of the bacteri...
Pinn-Woodcock TL, Aprea MS, Lejeune M, Tomlinson JE.A common diagnostic approach for febrile horses is to test for a panel of potential pathogens. Panels are curated by expert opinion and vary between diagnostic laboratories. Objective: To report the results of a newly developed equine fever diagnostic panel (EFDP) between 2019 and 2023 and evaluate the frequency of positive results. Methods: Retrospective descriptive study. Methods: The EFDP requires submission of whole blood, nasal swab, and faecal samples, and includes PCR tests for 12 pathogens that can present as fever without localising signs of illness or infection. Submission metadata a...
B B, G M, L G, G A, B B, T F, A G, D B, A K, G T, G S, A B, M F, L R.Equine penile tumors are common in horses and are often related to infection with equine papillomavirus type 2 (EcPV2). This study investigated the immune cell infiltrate (ICI) of these tumors in horses, focusing on the role of EcPV2. Using multiplex immunohistochemistry (mIHC) for CD3, CD20, and IBA-1 and immunohistochemistry (IHC) for FoxP3, 27 horses with papillomas (5/27), in situ carcinomas (CISs) (3/27), and squamous cell carcinomas (SCCs) (19/27) were evaluated. Eighteen cases tested positive for EcPV2 by either or both in situ hybridization (ISH) and polymerase chain reaction (PCR) (18...
Pádua GT, Tavares MA, de Lima NJ, Paula WVF, Dos Santos GC, Neves LC, Bittencourt RBM, Paludo RLDR, Cardoso ERN, da Silva BBF, Pádua BR....This study sought to investigate the presence of anti- spp. antibodies in georeferenced serum samples from equids across all regions of the state of Goiás, while also presenting variables that indicate risk factors for the circulation of rickettsiae, and evaluating the presence of rickettsial DNA in ticks collected from equids and the surrounding environment in the municipalities of Uruaçu and Porangatu, located in the northern region of the state. A total of 1156 equid serum samples provided by the Goiás Agricultural Defense Agency (Agrodefesa) were analyzed for antibodies against 4 antig...
James K, Chappell DE, Craig B, Pariseau C, Wright C, van Harreveld P, Barnum S, Pusterla N.The purpose of this study was to determine any associations of EHV-2, EHV-5, and dual infection with EHV-2/-5 with demographic parameters, clinical signs, and coinfection with other common respiratory pathogens. Nasal swabs collected from 9737 horses were tested for EHV-2 and EHV-5, as well as EHV-1, EHV-4, EIV, , ERAV, and ERBV, by qPCR. Clinical signs and demographic parameters were recorded, and prevalence factors were evaluated for significance regarding EHV-2 and/or EHV-5 infection. Out of the 9737 horses in this study, 17.8% tested EHV-2-positive ( = 1731), 15.8% tested EHV-5-positive ( ...
Nikvand AA, Jalali SM, Bahrami S, Rahij Torfi H.Trypanosoma evansi (T. evansi) is a hemoprotozoan parasite affecting camels and equids, such as horses, mules, and donkeys, and is known to cause surra disease in these animals. Despite the worldwide distribution of T. evansi infections in equids, surra has not been reported in Arabian horses in Khuzestan Province for over 60 years. In September 2018, a 7-year-old Arabian mare was referred from a 10-horse farm in the suburbs of Ahvaz City. The mare presented with a history of weight loss, poor appetite, and proximity to a camel herd. Physical examination revealed a poor body condition score ...
Mikaiel T, Waller A, Foote A, Cardwell JM, Mitchell J, Priestnall SL.Streptococcus equi subspecies zooepidemicus (S. zooepidemicus) is a commensal opportunistic bacterium associated with outbreaks of equine respiratory disease alongside a diverse range of diseases in various species. The closely related Streptococcus equi subspecies equi (S. equi) is the causative agent of 'strangles', the most frequently diagnosed contagious equine disease. Despite differing clinical signs, the two subspecies share approximately 97 % DNA homology and respectively present serious equine health and welfare concerns. Currently there are few PCR assays targeting unique regions of...
Perkins GA, Wagner B, Rollins A, Sfraga H, Pearson E, Cercone M.To apply equine herpesvirus type 1 (EHV-1) antibody testing in nasal swabs and serum in nonclinical horses during a naturally occurring outbreak of (EHV-1). Previous experimental EHV-1 challenge studies showed stable serum anti-EHV-1 antibody concentrations paired with rapidly increasing nasal mucosal antibodies (mucAbs) prevent EHV-1 infection, viral shedding, and cell-associated viremia. From this, we hypothesized that EHV-1 antibody testing can confirm exposure in non-clinical horses during an outbreak. Unassigned: 2 horses with neurological signs from 1 farm were admitted to an equine hosp...
Duaso J, Perez-Ecija A, Navarro A, Martínez E, De Las Heras A, Mendoza FJ.Equine piroplasmosis (EP) is an endemic parasitic disease in southern European countries, such as Spain. Andalusia, the most southwestern region of Europe, is the community with the highest number of registered horses and farms in Spain and one of the main international exporters of Andalusian (Spanish Purebred) horses worldwide. Considering the current expansion of this disease and the possible effect of climate change on its prevalence, studying the EP prevalence in this region is compelling. Molecular (PCR) and serological methods (cELISA and IFAT) were used to study the true and apparent p...
Kholik K, Sukri A, Priscilia Riwu KH, Ayu IW, Dewi IN.The transmission of () containing virulent genes from animals to humans and the environment poses significant public health challenges. This study aimed to detect the virulence factor of the () in isolated from the feces of apparently healthy horses in the island of Sumbawa, Indonesia. The study utilized 52 fecal samples from a total horse population of 283, calculated using the disease detection formula. Fresh feces were collected immediately after excretion and placed in buffered peptone water for subsequent analysis. The samples were then isolated on eosin methylene blue media and identi...
Kirmse L, Thieme K, Doherr MG, Eule JC.To evaluate different laboratory procedures for determining the etiologic diagnosis of equine recurrent uveitis regarding intraocular infection with Leptospira spp. and to establish a diagnostic guideline. Methods: Eighty horses with a history of ERU were ophthalmologically examined. Serum and aqueous humor were collected. Total protein, albumin level, and MAT against Leptospira spp. were evaluated on serum and aqueous humor. PCR for Leptospira spp., EHV-1 and -4 was performed on aqueous humor. Goldmann-Witmer coefficient (GWC) and C-value (CC) were calculated based on MAT. In 42 cases, an add...
Calero IM, McKenzie EC, Johns JL.This report describes the diagnosis and successful management of a yearling filly with Coombs-positive anemia, thrombocytopenia, and fungal pneumonia. Diagnostic procedures, including thoracic ultrasonography and radiography, respiratory pathogen PCR testing, and evaluation of tracheal wash and bronchoalveolar lavage samples established multi-pathogen lower respiratory tract infection including a fungal agent. Orally administered voriconazole was a key component of treatment in this case to successfully eliminate fungal infection, alongside therapies for managing hematologic disease. This case...
Cooper CJ, Arroyo LG, Hammermueller JD, Botts MM, Pearl DL, Wootton SK, Lillie BN.Equine herpesvirus-1 (EHV-1) is ubiquitous in the horse population, but prevalence estimates have ranged from 3 to 88% depending on the population and method of sampling. No prevalence studies have been carried out in Ontario, Canada. The objective of this study was to measure the prevalence of EHV-1 shedding in healthy broodmares in Ontario. A total of 381 mares from 42 farms in Ontario were sampled, including pregnant and barren broodmares. Samples were collected from the nose, vagina, and blood of each mare up to 6 times from December 2016 through October 2017 using a cross-sectional study ...
Ramírez-Agámez L, Castaneda C, Hernández-Avilés C, Grahn RA, Raudsepp T, Love CC.Two methods for preimplantation genetic testing (PGT) have been described for equine embryos: trophoblast cell biopsy (TCB) or blastocoele fluid aspiration (BFA). While TCB is widely applied for both in vivo- and in vitro-produced embryos, BFA has been mostly utilized for in vivo-produced embryos. Alternative methods for PGT, including analysis of cell-free DNA (CFD) in the medium where in vitro-produced embryos are cultured, have been reported in humans but not for equine embryos. In Experiment 1, in vivo- (n = 10) and in vitro-produced (n = 13) equine embryos were subjected to BFA, cultu...
Parashar R, Singla LD, Kaur P, Sharma SK.Relative association of haemato-biochemical findings with oxidative stress markers was evaluated between natural patent and latent infection of in horses to divulge the role of these parameters in the pathogenesis of illness due to non-availablity in literature. Blood samples were collected from 429 equines of 16 districts of the Punjab and samples positive by conventional microscopy (patent Group I; oll = 13), by polymerase chain reaction (PCR) (latent group II; = 38) and healthy control (group III, = 64) were compared for haematological-biochemical index and stress parameters....
Wickenden H, Lightbody KL, Peczak N, Stevens KB, Pollard D, Blake DP, Austin CJ, Matthews JB, Fox MT.Anoplocephala perfoliata is the most common equine tapeworm infection. This parasite is found at the small/large intestinal junction and has been associated with colic. The cestode has an indirect lifecycle involving oribatid mite intermediate hosts, though little is known of its epidemiology. This study aimed to monitor seasonal fluctuations in pasture oribatid mite numbers and the presence of Anoplocephala spp. DNA in mite samples collected from three equine premises in the UK. Exposure to infection in resident horses was assessed by measuring tapeworm-specific salivary antibodies. The data ...
Motta D, Pedrosa J, Lilenbaum W.Leptospirosis is a zoonotic disease caused by pathogenic bacteria of the genus Leptospira. A lesser-known form, equine genital leptospirosis (EGL), has been identified as a chronic and often silent infection involving the colonisation of the mare's genital tract. Despite its potential impact, EGL remains underdiagnosed and poorly understood, particularly in its association with reproductive inefficiency. This study showed the presence of Leptospira spp. DNA by lipL32-PCR in the genital tract of mares with a history of reproductive disturbances. Cervicovaginal mucus samples were collected from ...
Jelocnik M, Hall C, Dennis S, Mitchell K, Blishen A, Mashkour N, Anstey SI, Jenkins C, Jeffers K, El-Hage C, McMillan D, Gilkerson J.Infectious diseases significantly impact equine health and welfare, causing illness and death, and loss of productivity globally. One such disease is 'strangles', a highly contagious upper respiratory condition in horses caused by Streptococcus equi subspecies equi (SEE). Diagnostic methods for this pathogen include sensitive molecular assays and less reliable bacterial isolation and biochemical testing. However, the presence of closely related streptococci, such as Streptococcus equi subspecies zooepidemicus (SZOO), may confound diagnosis. Rapid assays for SEE are crucial for outbreak control...
Kambayashi Y, Bannai H, Nemoto M, Kawanishi N, Niwa H, Tsujimura K.With the revision of the World Organisation for Animal Health (WOAH) Terrestrial Manual on equine rhinopneumonitis in 2024, 3 recommended qPCR primer-probe sets were added for the detection of equid alphaherpesvirus 1 (EqAHV1; formerly equine herpesvirus 1 [EHV1]; family , taxon species ), also known as equine abortion virus. We compared the sensitivity and specificity of the 3 qPCR primer-probe sets to determine the most reliable set. Sets gB1H and gB1P, which target the glycoprotein B () gene of EqAHV1, detected all 10 copies and even lower copy numbers. In contrast, set gC1 (ISO 17025-accre...
Jamshidpour R, Nabavi R, Moadab H, Rezaie F, Chale AC, Sargison N.Resistance to benzimidazole (BZ) by cyathostomin nematodes has become a major threat to equine health around the world. We aimed to evaluate the efficacy of BZ drugs against small strongyle nematodes in horses of western Iran using coprological and molecular examination. Unassigned: Faecal egg count reduction tests were performed on 398 horses kept in 16 stables in western Iran (Kermanshah Province), to detect benzimidazole resistance in small strongyle nematodes. Allele-specific PCR was used to identify the F200Y (TAC/TTC) SNP in the beta-tubulin gene codon in cyathostomin larvae. Unassigned:...
Zanilabdin M, Ilgekbayeva G, Otarbayev B, Nissanova R, Mussayeva G, Takai S, Suzuki Y, Kakuda T, Kurman S, Kassymov Y, Valiyeva B. is a facultative intracellular pathogen causing bronchopneumonia in foals; data from Central Asia are limited. We conducted a cross-sectional serological and molecular survey in horses from three regions of Kazakhstan (Kyzylorda, Almaty, Akmola). Unassigned: Sera from 312 animals (272 adults, 40 foals) on 20 farms were tested by indirect ELISA. Selected clinical samples underwent culture, PCR, and 16S rRNA sequencing. Unassigned: Overall seroprevalence was 8.3% (26/312; 95% CI 5.8-11.9). Positivity among foals was 25.0% (10/40; 95% CI 14.2-40.2) versus 5.9% (16/272; 95% CI 3.7-9.3) in adults,...
McLachlan AD, Woolford L.Chlamydia psittaci was detected by real-time PCR in the lung, liver and kidney of an equine foetus that had aborted in South Australia in August 2023. The corresponding microscopic lesions included lymphocytic and histiocytic chorionitis, necrosis of placental villi associated with bacteria in the cytoplasm of trophoblastic epithelial cells, and multiple microgranulomas in the liver. Equine chlamydial abortion had not been diagnosed previously in South Australia. Eight days after examining the foetus and placenta, the veterinary pathologist developed fever and subsequently was admitted to hosp...
Alsultan A, Karim SM, Al-Saadi M, Alsallami D, Ben Said M, Belkahia H.Equine piroplasmosis (EP), caused by the intracellular protozoa Theileria equi, Babesia caballi, and Theileria haneyi, represents a major health and economic threat to the equine industry worldwide. Existing diagnostic methods, including PCR, serology, and microscopy, are constrained by their dependence on specialized equipment, lengthy protocols, and the requirement for skilled personnel. Objective: This study aimed to develop a rapid, accurate, and field-deployable molecular diagnostic assay for T. equi. Methods: A nucleic acid-based diagnostic platform combining recombinase polymerase ampli...
