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.
Potgieter FT, de Waal DT, Posnett ES.The transmission and prevalence of Babesia equi and B. caballi are being studied. Rhipicephalus evertsi mimeticus an ixodid tick from Namibia was identified as a new vector of B. equi, however, R. turanicus, previously reported to be a vector, failed to transmit both B. equi and B. caballi in the laboratory. The accurate diagnosis of B. caballi is being investigated because the nature of its low level parasitaemia does not allow easy detection in thin blood smears, routinely used for diagnosis, by clinicians. Consequently its role as a pathogen remains obscure. The importance of identifying in...
Sharma PC, Cullinane AA, Onions DE, Nicolson L.The polymerase chain reaction (PCR) is a sensitive technique used to detect DNA of viral pathogens. We have applied the technique to the detection of Equid herpesviruses-1 and -4 (EHV-1 and EHV-4) DNA within nasopharyngeal swab samples from horses. Ninety-eight samples from suspected field cases and in-contact horses were analysed. The assays were conducted blind and later decoded and compared with virus isolation data. Our results indicate that PCR is a sensitive and rapid technique for the diagnosis of EHV-1 and EHV-4 infection.
Ellegren H, Johansson M, Sandberg K, Andersson L.We have isolated equine microsatellites by screening a genomic library with (TG)n and (TC)n probes. TG microsatellites were found to be more abundant than TC repeats, with an estimated frequency of one per 100,000bp. Sequence analysis of eight TG-positive clones revealed varying structures of the repeat regions; perfect stretches of TG repeats, imperfect stretches of TG repeats and compound regions of TG and TC repeats. Five loci were analysed by PCR and showed extensive polymorphism; three to seven alleles and heterozygosities of 0.40-0.76 were observed when screening 20-30 unrelated individu...
Su XZ, Morris DD, McGraw RA.We report the molecular cloning and nucleotide sequence of the equine gene encoding tumor necrosis factor alpha. The 2610-bp genomic sequence was derived from three overlapping polymerase chain reaction products.
Biswas B, Mukherjee D, Mattingly-Napier BL, Dutta SK.Genomic amplification by the polymerase chain reaction (PCR) was used to identify a unique genomic sequence of Ehrlichia risticii directly in DNA isolated from peripheral-blood buffy coat cells of E. risticii-infected horses (Potomac horse fever) and from infected cell cultures. A specific primer pair, selected from a cloned, species-specific, 1-kb DNA fragment of the E. risticii genome as a template, was used for the amplification of the target DNA of 247 bp. The optimal number of 40 PCR cycles, determined by analyzing an amplification profile obtained with a constant Taq polymerase concentra...
Alexandersen S, Carpenter S.The polymerase chain reaction was used to amplify and clone parts of the envelope gene and overlapping S3 open reading frame, thought to encode rev, of the virulent in vivo-derived Th-1 isolate of equine infectious anemia virus (EIAV). The results indicated that EIAV consists of a heterogeneous mixture of genotypes present at the first febrile cycle after initial infection. We showed that the Th-1 isolate apparently contains nondefective genotypes as well as types which have transmembrane protein truncations or are rev deficient. Furthermore, we could confirm the presence of a hypervariable re...
Teifke JP, Weiss E.Unfixed and formalin-fixed frozen sections and paraffin-sections of histopathologically confirmed sarcoids of 20 horses were studied in the PCR. The used set of primers was located in the E5 open reading frame fitting both to bovine papillomavirus 1 (BPV-1) and BPV-2. Independent of the quality of the used tissues BPV-DNA was detected in all 20 sarcoids. By cleaving with restriction endonuclease Bst XI it was shown that the DNA-sequences amplified by PCR were identical with that of BPV-1. The results support the general view that BPV play an important role in equine sarcoids.
O'Keefe JS, Murray A, Wilks CR, Moriarty KM.Unpurified DNA derived from cultures of equine fetal kidney cells infected with either equine herpesvirus type 1 or equine herpesvirus type 4 was amplified by the polymerase chain reaction using one pair of oligonucleotide primers. Restriction endonuclease digestion of the amplified segments with PvuII, followed by electrophoresis, revealed restriction fragment length polymorphisms which enabled the two virus types to be differentiated.
O'Rourke KI, Besola ML, McGuire TC.Proviral sequences in the peripheral blood mononuclear cells of 3 horses with acute equine infectious anemia virus were monitored using the polymerase chain reaction. Provirus was detected during the initial viremic episode in each horse and during each of 3 relapsing viremic cycles, although the appearance of provirus lagged behind the onset of viremia. Following each viremic episode, provirus levels in the peripheral monocytes decreased to less than 1 copy in 5 x 10(6) cells.
Whetter L, Archambault D, Perry S, Gazit A, Coggins L, Yaniv A, Clabough D, Dahlberg J, Fuller F, Tronick S.A full-length molecular clone of equine infectious anemia virus (EIAV) was isolated from a persistently infected canine fetal thymus cell line (Cf2Th). Upon transfection of equine dermis cells, the clone, designated CL22, yielded infectious EIAV particles (CL22-V) that replicated in vitro in both Cf2Th cells and an equine dermis cell strain. Horses infected with CL22-V developed an antibody response to viral proteins and possessed viral DNA in peripheral blood mononuclear cells, as determined by polymerase chain reaction assays. In addition, horses infected with CL22-V became persistently infe...
Ballagi-Pordány A, Klingeborn B, Flensburg J, Belák S.Primers and probes were selected from the gene encoding glycoprotein 13 (gp 13) of equine herpesvirus 1 (EHV-1). The polymerase chain reaction (PCR) was run on infected and noninfected cultured cells and on 63 specimens from 29 aborted equine fetuses. The results were evaluated by electrophoresis and dot-blot hybridization using an oligonucleotide probe labeled with biotin. In the infected samples electrophoresis showed a PCR product of about 280 base pairs. The dot-blot hybridization confirmed that this product contained EHV-1 DNA sequences. PCR took 4 h and hybridization another 14 h; the re...
Pomelova VG, Gaĭdamovich SIa, Demenev VA, Kadoshnikov IuP.A three-step concentration of Venezuelan equine encephalomyelitis (VEE) virus from tissue culture fluid was carried out in a two-phase system of polyethyleneglycol (PEG)--sodium dextran sulphate (SDS). The concentration method was based on the dependence of virus distribution coefficient upon NaCl content in the system which allowed alternating transfer of the virus from one phase of the system into the other. The infectious activity of the virus increased approximately 100-fold after the first step, 190-fold after the second, and 300-fold after the third step. The process of concentration was...
Skok MV, Denisiuk PV, Komissarenko SV.Glutaraldehyde treatment does not change the absorption of cytochrome c either in the visible or in UV spectra. It brings about the formation of dimers, trimers and high-polymeric forms of cytochrome c and shifts the pI of all cytochrome c isoelectric fractions to more acid pH. Polymerization also results in changes of kinetic parameters of cytochrome c benzidine reaction increasing its affinity to 3,3-diaminobenzidine with a simultaneous decrease in the effectiveness of H2O2 binding. These biochemical changes can be related to immunochemical differences of native and glutaraldehyde-treated cy...
The goal of this work was the development of suitable (real-time) RT-PCR techniques for fast and sensitive diagnosis of EAV and for molecular-epidemiological characterisation of viral strains, as an alternative to virus isolation. To this purpose two conventional RT-PCR methods and one real-time RT-PCR were adapted to detect the broadest possible spectrum of viral strains. Several dilutions with Bucyrus strain showed a 100-fold higher sensitivity of real-time RT-PCR and heminested RT-PCR compared to simple RT-PCR. Making use of 11 cell culture supernatants of different EAV isolates and 7 semen...
Kamiński S, Bejda J, Lewczuk D.The method for identifying the causative mutation for Warmblood Fragile Foal Syndrome (WFFS) involved PCR amplification of a 259-base pair fragment of the PLOD1 gene and its digestion with the restriction enzyme Aci I was developed, allowing for the clear detection of WFFS carriers. Eight WFFS carriers were detected among 308 warmblood horses kept in different farms across Poland, giving an overall frequency of 2.59%, which indicates a rather low frequency of the causative mutation for WFFS in Poland. Further research should be conducted on a larger number of horses, particularly those breeds ...
Gao W, Liu M, Nurdaly K, Caidan D, Sun Y, Duan J, Zhao J, Gong X, Zhou J, Zhang Y, Chen Q.Equine bacterial abortion presents substantial economic and One Health challenges; however, comprehensive epidemiological data from China are limited. This study sought to ascertain the overall prevalence of key pathogens-namely, spp., , , and spp.-in equine populations in northwestern China. In this study, we aimed to further elucidate the characteristics of co-infections, profile antimicrobial resistance genes, and identify associated risk factors. Conducted as a cross-sectional analysis across four provinces, we collected 508 blood samples and 24 abortion tissue samples from 15 farms. Pat...
Sfraga H, Demeter EA, Pinn-Woodcock T, Guarino C, Young R, Cronk B, Cercone M.To investigate the presence of subclinical cranial nuchal bursitis and characterize its histopathologic features and association with Borrelia burgdorferi. Unassigned: This was a prospective descriptive cadaver study on a convenience population of horses in a B burgdorferi-endemic region (15 horses: 5 geldings and 10 mares of various breeds; 4 to 29 years old). Horses without history or clinical signs of cranial nuchal bursitis underwent euthanasia and tissue donation. Cranial nuchal bursa, synovial fluid, and nuchal ligament were collected postmortem. The bursa and ligament were evaluated via...
Penzhorn L, Crafford JE, Guthrie AJ.African horse sickness (AHS) is the only equine disease for which the World Organisation for Animal Health (WOAH) gives official disease-free status, given that it poses a major threat to the equine industry. The disease is caused by AHS virus (AHSV; family , taxon species ), which is endemic in sub-Saharan Africa. Reverse-transcription quantitative real-time PCR (RT-qPCR) is a rapid, sensitive detection method used in the diagnosis of AHS and the certification of animals as negative for AHSV for the purpose of movement. Genetic variability of AHSV may influence the accuracy of RT-qPCR detecti...
Dorrego A, Olvera-Maneu S, Jose-Cunilleras E, Gago P, Raez A, Rivera B, Oporto A, Gonzalez S, Cruz-Lopez F.The forest fly ) is an obligate haematophagous dipteran insect (order Diptera) that primarily infests horses and may contribute to the circulation of vector-borne pathogens. This study aimed to investigate the presence of , s.l., , and , important vector-borne pathogens of equids, in forest flies collected from horses in endemic areas of Spain. A total of 170 forest flies were collected from 39 equids across four geographical regions in Spain (Segovia, Madrid, Toledo, and Menorca) and blood samples were collected from 27 of these horses. All flies were morphologically and molecularly identifi...
Toda J, Miyasaka J, Osako H, Murata K, Yunus M, Amalia R, Soe BK, Sato H.Food poisoning caused by consuming raw horsemeat contaminated with Sarcocystis is a significant public health concern. Two morphotypes of sarcocysts in horsemeat, characterized by upright and folded villar protrusions, are typically identified as Sarcocystis fayeri and S. bertrami, respectively. However, recent molecular studies focusing on the ribosomal RNA gene (rDNA) and mitochondrial cytochrome c oxidase subunit I gene (cox1) have indicated a conspecific relationship between these two morphotypes using a limited number of specimens. To explore further genetic diversity in equid sarcocysts,...
Davoudi N, Behbahani M, Mohabatkar H, Dini G, Bakhshesh M.Equine herpesvirus type 1 (EHV-1) is a globally prevalent equine pathogen responsible for severe respiratory, neurological, and reproductive disorders. Accurate and ultrasensitive detection of EHV-1 is critical for timely disease management. In this study, we report the development of the first G-quadruplex-forming aptamer specifically designed for EHV-1 detection. The aptamer was generated using an in silico approach, and its G-quadruplex conformation was confirmed using circular dichroism (CD) spectroscopy and crystal violet fluorescence assays. Binding affinity and specificity were assessed...
Tramboo SR, Shahardar RA, Allaie IM, Bulbul KH.Benzimidazole (BZ) anthelmintics have been used indiscriminately in equids to control nematode infections throughout the world including India and has led to the development of BZ resistance. In order to determine the current status of BZ resistance in equids of Kashmir against intestinal strongyles (IS), the present study was conducted using faecal egg count reduction test (FECRT) and Allele specific PCR (AS-PCR). The study was conducted on ponies from three major tourist destinations of Kashmir viz; Gulmarg, Pahalgam and Sonamarg in accordance with the WAAVP guidelines. The animals which wer...
Morales J, Ruano MJ, Tena-Tomás C, van Schalkwyk A, Loundras EA, Valero-Lorenzo M, López-Herranz A, Romito M, Batten C, Villalba R, Agüero M.African horse sickness (AHS) is a disease affecting equids caused by the AHS virus (AHSV). The World Organisation for Animal Health (WOAH) includes AHS as a notifiable disease and, upon detection within the European Union, immediate control and eradication measures are mandated. Thus, validated diagnostic methods for rapid AHSV detection are essential. The Agüero 2008 and Guthrie 2013 rRT-PCR methods have been widely validated for detection of any AHSV strain and are included as reference rRT-PCRs in the WOAH manual. However, the WOAH Reference Laboratory for AHS in the Republic of South Afri...
van Rijn PA, Wernery U, Feddema AJ, Maris-Veldhuis MA, Joseph S, van Gennip RGP.African Horse Sickness (AHS) is a devastating vector-borne viral disease of equids with a mortality up to 95 % in naïve domestic horses. The causative African horse sickness virus (AHSV) is a distinct species of the genus Orbivirus of the family Sedoreoviridae, consisting of nine serotypes showing limited cross protection. AHSV is transmitted by Culicoides biting midges. Outbreaks cause huge economic losses in developing African countries. AHS has become a serious threat for countries outside Africa, since endemic Culicoides species in moderate climates appear competent vectors of the closel...
Snedden K, Frye E, Conklin R, Aprea M, Rishniw M, Lejeune M, Goodrich E.To analyze the results and metadata of canine, feline, and equine respiratory PCR panel assays performed at the New York State Animal Health Diagnostic Center and inform veterinary diagnostic sample submission. Unassigned: This retrospective study reviewed laboratory data from routine sample submissions to the Animal Health Diagnostic Center for canine, feline, and equine respiratory PCR panels from January 1 through December 31, 2023. Associations were compared between variables using χ2 tests of independence or Fisher exact tests. Unassigned: A total of 1,902 canine, feline, and equine resp...
Molazadeh S, Tukmechi A, Hadian M, Dalir-Naghadeh B.This study aimed to determine the prevalence and phylogenetic analysis of Ehrlichia spp. in horses and dogs in Iran. Blood samples were collected from 400 animals, including 200 horses and 200 dogs, from five different provinces in Iran. Polymerase chain reaction (PCR) was used to detect Ehrlichia spp. based on amplification of the 16S rRNA gene. The semi-nested PCR method was used to amplify the dsb, TRP36, and gltA genes. The results showed that 4.5 % of the samples (3 % horses and 6 % dogs) were positive for Ehrlichia sp. The highest prevalence was observed in Kerman and Khuzestan, while th...
Silva-Ramos CR, Niño Rodríguez JA, Gil-Mora J, Betancourt-Ruiz P, Martínez-Díaz HC, Forero-Becerra E, Matiz-González JM, Bolaños E, Olaya-M LA....Babesia species are tick-borne protozoan parasites which affect several animal species. Babesia spp. infections are significantly important for veterinary medicine, affecting a wide range of domestic animal species such as dogs, cattle, and horses. In Colombia, studies of Babesia spp. infections in domestic animals are scarce. Thus, the aim of the present study was to explore the circulation of these parasites among domestic canines, bovines and equines from the department of Cauca. Methods: Between August and November, 2017, active domestic animal sampling of cattle was performed in eight rur...
Sousa JA, Miranda LM, Coutinho DJB, Costa TF, Costa SP, Freitas ÚS, Costa FB, Machado RZ, Nogueira RMS, Costa APD.The hemoprotozoan Trypanosoma evansi is a parasite that infects mammals, causing an infection known as trypanosomiasis. There is no report of T. evansi in horses in the State of Maranhão, highlighting the need to assess exposure and infection by the parasite and generate data for its monitoring. The objectives of this study were to identify T. evansi in blood samples from horses, investigate its occurrence in horses in this region, and analyze the associated risk factors. Three hundred blood samples were collected for parasitological (blood smear), serological (indirect enzyme-linked immunoso...
Charbonnel A, Lavoie JP, Juette T, St-Sauveur VG, Denis S, Gagnon CA, Leclère M.The control of equine respiratory infections is a biosecurity challenge. Respiratory viruses are often rapidly detected using quantitative polymerase chain reaction (qPCR) on nasal swabs. In the past, some laboratories developed handmade techniques to increase the amount of nasal secretions collected, without comparing them with nasal swabs when qPCR replaced the use of viral culture. The objectives of this study were to compare nasal swabs and handmade foam cubes for i) the detection of a common equine herpesvirus (EHV-5) by qPCR, and ii) their tolerability. Forty-five polyester swabs and foa...
Olguin-Perglione C, Politzki R, Alvarez I, Ruiz V.The novel Equine Parvovirus-Hepatitis (EqPV-H) was first identified in the serum and liver of a horse that died of equine serum hepatitis, also known as Theiler's disease. Several reports in recent years strongly suggest that EqPV-H is the etiologic agent of Theiler's disease. Brazil is the only South American country where infection with this virus has been reported. This study investigated the presence of EqPV-H DNA in horse serum pools (n=51), commercial horse serum batches (n=5) and individual serum samples from donor horses (n=175) from Argentina. All serum samples were analyzed by quanti...
Lale D, Dirks EE, Preining I, Lyrakis M, Gömer A, Steinmann E, Cavalleri JV, Ramsauer AS.Equine parvovirus hepatitis (EqPV-H) can cause Theiler's disease and subclinical hepatitis in horses. Objective: Assess the frequency of subclinical EqPV-H infection in hospitalized horses and to study viral transmission by investigating potential shedding routes. Methods: One hundred sixteen equids, that presented to the University Equine Hospital of the University of Veterinary Medicine Vienna between February 2021 and March 2022, for causes other than hepatopathy. Methods: In this cross-sectional study, samples (serum, feces, nasal, and buccal swabs) of hospitalized horses were collected. S...
Cutarelli A, Buonavoglia A, Fusco G, Pellicanò R, Napoletano M, Brandt S, Roperto S.Sarcoids are benign and locally aggressive skin lesions that commonly affect horses and other equid species. Sarcoids are generally considered to be caused by bovine delta-papillomaviruses (δPVs) types 1 and 2 (BPV1 and BPV2, respectively). Moreover, while bovine δPV types 13 and 14 (BPV13 and BPV14, respectively) are also suspected to induce sarcoids, information regarding this possibility and the occurrence of multiple bovine δPV infections in sarcoids is scarce. This study aimed, for the first time, to assess BPV1, BPV2, BPV13, and BPV14 infections and co-infections in equine sarcoid sam...
