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.
Fortier G, Pronost S, Miszczak F, Fortier C, Léon A, Richard E, Van Erck E, Thiry E, Lekeux P.During a case control study undertaken in 2006-2007, a screening and consensus polymerase chain reaction (PCR) was performed to evaluate the potential role of equid herpesviruses (EHV) in several occurrences of respiratory disorders in 661 horses. Of 785 bronchoalveolar or tracheal lavage fluid samples submitted for analysis, 20 were positive for EHV-5 DNA by sequential analysis of the consensus PCR product. Nineteen of those samples were confirmed using a specific EHV-5 PCR. No particular changes in cytological profile could be associated with the detection of EHV-5 in contrast to suggestions...
Díaz S, Echeverría MG, It V, Posik DM, Rogberg-Muñoz A, Pena NL, Peral-García P, Vega-Pla JL, Giovambattista G.The polymorphism of equine lymphocyte antigen (ELA) class II DRA gene had been detected by polymerase chain reaction-single-strand conformational polymorphism (PCR-SSCP) and reference strand-mediated conformation analysis. These methodologies allowed to identify 11 ELA-DRA exon 2 sequences, three of which are widely distributed among domestic horse breeds. Herein, we describe the development of a pyrosequencing-based method applicable to ELA-DRA typing, by screening samples from eight different horse breeds previously typed by PCR-SSCP. This sequence-based method would be useful in high-throug...
Perkins GA, Goodman LB, Dubovi EJ, Kim SG, Osterrieder N.Early identification of inhalation-transmitted equine herpesvirus type 1 (EHV-1) infections has been facilitated by the availability of a number of real-time quantitative PCR (qPCR) tests. A direct comparison between nasal swab qPCR and traditional virus isolation (VI) requires a method for normalizing the qPCR samples and controlling for PCR inhibitors present in some clinical samples. Objective: To quantify EHV-1 shedding in viral swabs using an internal control and to compare fast qPCR to VI for the detection of EHV-1 in nasal swabs from horses. Methods: Fifteen horses experimentally infect...
Frazer ML.Lawsonia intracellularis is an emerging equine pathogen that is a cause of equine proliferative enteropathy (EPE). Objective: To describe the signalment, month of presentation, common clinical signs, clinicopathologic values, diagnostic tests used, antimicrobial use, and survival status in horses affected with EPE; to evaluate how affected horses sold at public auction as yearlings; and to determine results of fecal polymerase chain reaction (PCR) and serum immunoperoxidase monolayer assay (IPMA) results in age matched, clinically normal herdmates. Methods: The study group was 57 horses treate...
Steel CM.The most important application for synovial fluid (SF) analysis in the horse is in the diagnosis of synovial sepsis. Misdiagnosis of synovial sepsis is costly, and SF analysis makes correct diagnosis more likely, although far from certain. The precision of diagnosis may be increased with polymerase chain reaction analysis for detection of bacterial DNA in SF and with assays for various enzymes and cytokines. These tests are currently not widely available, however, and routine SF analysis remains of prime importance in diagnosis.
Koblischke P, Kindahl H, Budik S, Aurich J, Palm F, Walter I, Kolodziejek J, Nowotny N, Hoppen HO, Aurich C.We tested the hypothesis that subclinical endometritis occurs after embryo transfer (ET) in the horse. Recipient mares were treated with meclofenamic acid (M) or flunixin meglumin (F) after ET or were left untreated (n=9 per group). Embryos were re-collected 4 days after transfer. Endometrial biopsies were taken for histology and analysis of cyclooxygenase-2 (COX-2) by immunohistochemistry and for PCR. Bacteriological swabs were collected from the uterus and lavage fluid of donor and recipient mares. Progesterone and prostaglandin F(2alpha) release was analysed in recipient mares after ET. Fou...
Zoonoses and public healthJuly 22, 2008
Volume 55, Issue 6 291-298 doi: 10.1111/j.1863-2378.2008.01136.x
Zhang W, Shen Q, Mou J, Gong G, Yang Z, Cui L, Zhu J, Ju G, Hua X.Hepatitis E virus (HEV) is a zoonotic pathogen of which several species of animal were reported as reservoirs. Antibodies to HEV and HEV RNA have been detected in some Chinese population and swine groups but few other domestic animals. In this study, to investigate the HEV prevalence, we tested sera from 788 pigs, 100 cows, 50 goats, 49 horses, 101 pet dogs, 105 chickens, 47 duck and 45 pigeons in eastern China for anti-HEV immunoglobulin G (IgG). We also tested 50% of the swine sera, all of sera from the other domestic animals and 13 Shanghai human sera which were positive for anti-HEV immuno...
Ataseven VS, Dağalp SB, Güzel M, Başaran Z, Tan MT, Geraghty B.In this report we examined the presence of specific antibodies against equine herpesvirus type 1 (EHV-1), and equine herpesvirus type 4 (EHV-4) in several equidae, including mules, donkeys, horses. The presence of EHV-1 and EHV-4 in respiratory diseases of equids, and ability of multiplex nested polymerase chain reaction (PCR) screening in simultaneous diagnosis of horses acutely infected by EHV-1 and EHV-4 were also investigated. Sera from 504 horses, mules and donkeys sampled were tested for the presence of EHV-1 and EHV-4 specific antibodies. Blood samples taken from 21 symptomatic horses a...
Salim BO, Hassan SM, Bakheit MA, Alhassan A, Igarashi I, Karanis P, Abdelrahman MB.The purpose of this study was to estimate the prevalence of equine piroplasmosis in Sudan. The presence of antibodies against Babesia caballi and Theileria equi was determined in serum samples obtained from 158 horses raised in different locations in Sudan by enzyme-linked immunosorbent assay (ELISA). The B. caballi 48-kDa and the T. equi EMA-2 purified recombinant proteins were used as antigens in the ELISA test. Results showed that seven (4.4%) were positive for B. caballi and 80 (63.5%) were positive for T. equi. Polymerase chain reaction (PCR) assays have been applied using primers targeti...
Davidson A, Traub-Dargatz JL, Magnuson R, Hill A, Irwin V, Newton R, Waller A, Smith K, Callan RJ, Meehan M, Owen P, Salman M.Previously published studies have neither used nor reported the results of an indirect enzyme-linked immunosorbent assay (iELISA) to measure serologic responses in natural outbreaks of strangles. The concept of using serologic responses to identify persistent carriers of Streptococcus equi has been proposed but not scientifically evaluated. The specific aims of the current study were to determine the duration and level of truncated fibrinogen-binding protein-specific (SeM allele 1) antibody production in ponies involved in a natural outbreak of strangles and to determine if test results from t...
Menozzi A, Pozzoli C, Poli E, Dacasto M, Giantin M, Lopparelli RM, Passeri B, Zullian C, Gobbetti T, Bertini S.We investigated the effects of nonselective cyclooxygenase (COX) inhibitors (indomethacin and flunixin meglumine) and selective COX-1 (SC-560) or COX-2 (celecoxib, DUP-398 and NS-697) inhibitors on horse small bowel motility in vitro. At this purpose, samples of equine ileum were put in isolated organ baths for the motility experiments. Nonselective COX inhibitors were devoid of major effects on motility, except for an inhibition of tonic contraction shown by flunixin meglumine. SC-560, selective COX-1 inhibitor, was devoid of significant effects on ileal motility. Selective COX-2 inhibitors r...
Moyaert H, Haesebrouck F, Dewulf J, Ducatelle R, Pasmans F.Faecal samples of sixty-six 3-day- to 6-month-old foals were screened for Helicobacter equorum DNA by means of a PCR amplifying a 1074 bp fragment of the 23S rRNA gene with primers specific for this enterohepatic Helicobacter species. H. equorum DNA was demonstrated in faeces from 28.6% of the less than 1-month-old foals, while 67.8% of foals from 1 to 6 months of age tested positive. In a previous study, H. equorum was demonstrated in faeces of 0.8-7.9% of adult horses. These results indicate that the prevalence of H. equorum in horses differs with the age of the investigated horse population...
Turton JF, Baklan H, Siu LK, Kaufmann ME, Pitt TL.A multiplex PCR using targets within the serotype-specific region of the capsular polysaccharide synthesis gene cluster of serotypes K1, K2 and K5 was evaluated using the 77 reference serotype strains of Klebsiella, and a panel of clinical isolates subjected previously to conventional serotyping. The PCR was highly specific for these serotypes, which are those most associated with virulence in humans and horses. PCR confirmed that isolates of the K5 serotype had cross-reacted with antiserum for other serotypes, particularly for K7. K5 isolates received by our laboratory were almost exclusively...
Agüero M, Gómez-Tejedor C, Angeles Cubillo M, Rubio C, Romero E, Jiménez-Clavero A.African horse sickness is an arthropod-borne disease of the equine included in the World Organization for Animal Health (OIE) list with important economic consequences for horse trade. The disease is caused by African horse sickness virus (AHSV; family Reoviridae, genus Orbivirus), which is transmitted by Culicoides midges. It is endemic in sub-Saharan Africa, spreading occasionally outside this area where the occurrence of Culicoides vectors allows virus transmission. Currently, only conventional (gel-based) reverse transcription polymerase chain reaction (RT-PCR) protocols are available for ...
Hastie PM, Mitchell K, Murray JA.There is a need to further our understanding of the role that the equine hindgut ecosystem plays in digestive processes and diseases. The aim of the present study was to utilise the real-time PCR technique to determine the abundance of candidate cellulolytic (Ruminococcus flavefaciens; Fibrobacter succinogenes) and non-cellulolytic (Streptococcus bovis) bacteria in lumen contents from the caecum, ventral and dorsal colon, and rectum of healthy horses (n 14). Total DNA was extracted from frozen and lyophilised lumen contents, and PCR primers and Taqman probes were designed based on 16S rDNA seq...
Budik S, Walter I, Tschulenk W, Helmreich M, Deichsel K, Pittner F, Aurich C.Expansion of the equine conceptus can be divided into blastocoel and yolk sac phases. The endodermal layer transforming the blastocoel into the yolk sac is completed around day 8 of pregnancy. From that time, the size of the spherical conceptus increases tremendously due mainly to the accumulation of fluid rather than cell multiplication. In this study, we have investigated the abundance and localisation of Na(+)/K(+)-ATPases and aquaporins (AQP) in the equine conceptus on days 8, 10, 12, 14 and 16 by multiplex reverse transcriptase PCR, Western blot and immunohistochemistry. During conceptus ...
Marenzoni ML, Passamonti F, Cappelli K, Veronesi F, Capomaccio S, Supplizi AV, Valente C, Autorino G, Coletti M.Fifteen unweaned thoroughbred foals, born on a stud farm to vaccinated mares, were clinically monitored during their first six months of life and repeatedly tested for equine herpesvirus type 1 (EHV-1) and equine herpesvirus type 4 (EHV-4). Nasopharyngeal swabs and blood samples were collected and screened respectively by PCR and seroneutralisation to detect the presence of the virus, explore its role as a possible cause of respiratory disease, and to assess the efficiency of the pcr for the diagnosis of this disease. The foals were divided into three groups on the basis of their clinical sign...
Wong DM, Belgrave RL, Williams KJ, Del Piero F, Alcott CJ, Bolin SR, Marr CM, Nolen-Walston R, Myers RK, Wilkins PA.5 horses were evaluated because of decreased appetite, weight loss, fever, cough, tachypnea, and respiratory distress. Results: Tachycardia, tachypnea, increased respiratory effort, lethargy, fever, poor body condition, and nasal discharge were detected in various combinations on initial physical examination. Evaluation of the lower portion of the respiratory tract via radiography and ultrasonography revealed a severe nodular interstitial pattern. Histologic examination of lung tissue revealed interstitial expansion of alveolar parenchyma with collagen, intraluminal accumulation of neutrophils...
Torfason EG, Thorsteinsdóttir L, Torsteinsdóttir S, Svansson V.The horse population in Iceland is a special breed, isolated from other horses for at least 1000 years. This provides an exceptional opportunity to investigate old and new pathogens in an inbred herd with few infectious diseases. We have developed a high sensitivity semi-nested PCR to study equid gammaherpesviruses 2 and 5 (EHV-2 and 5) in Iceland. The first PCR is group specific, the second type-specific, targeting a 113bp sequence in the glyB gene. DNA isolated from white blood cells and 18 different organs was tested for the presence of EHV-2 and 5. This was done in adult horses and foals, ...
Jones SL, Maggi R, Shuler J, Alward A, Breitschwerdt EB.Bartonella spp. are emerging zoonotic agents that have been found in a wide variety of domestic animals and wildlife and cause a number of clinical syndromes. Bartonella sp. infection has been identified in a growing number of animal species, including cats, rodents, porpoises, and canids, but has not been reported in horses. Objective: To document the presence of Bartonella sp. in the blood of horses. Methods: One horse with chronic arthropathy and 1 horse with presumptive vasculitis. Methods: Blood samples were tested for the presence of Bartonella sp. by a combination of multiplex real-time...
Lu Z, Branscum AJ, Shuck KM, Zhang J, Dubovi EJ, Timoney PJ, Balasuriya UB.Two previously developed TaqMan fluorogenic probe-based 1-tube real-time reverse transcription polymerase chain reaction (real-time RT-PCR) assays (T1 and T2) were compared and validated for the detection of Equine arteritis virus (EAV) nucleic acid in equine semen and tissue culture fluid (TCF). The specificity and sensitivity of these 2 molecular-based assays were compared to traditional virus isolation (VI) in cell culture. The T1 real-time RT-PCR had a higher sensitivity (93.4%) than the T2 real-time RT-PCR (42.6%) for detection of EAV RNA in semen. However, the T1 real-time RT-PCR was les...
Rüegg SR, Heinzmann D, Barbour AD, Torgerson PR.For the evaluation of the epidemiology of Theileria equi and Babesia caballi in a herd of 510 horses in SW Mongolia, several mathematical models of the transmission dynamics were constructed. Because the field data contain information on the presence of the parasite (determined by PCR) and the presence of antibodies (determined by IFAT), the models cater for maternal protection with antibodies, susceptible animals, infected animals and animals which have eliminated the parasite and also allow for age-dependent infection in susceptible animals. Maximum likelihood estimation procedures were used...
Pinchbeck GL, Morrison LJ, Tait A, Langford J, Meehan L, Jallow S, Jallow J, Jallow A, Christley RM.The Gambia has an increasing population of equidae largely used for agriculture and transportation. A review of cases at The Gambian Horse and Donkey Trust (GHDT) indicated that a common reason for presentation is a poorly defined medical condition often attributed to trypanosomosis. There are few reports describing the prevalence or the range of clinical signs associated with infection with different species of trypanosomes in horses and donkeys, but given the importance of these animals, the role of trypanosomosis requires investigation. Results: In total 241 animals from the Central River D...
Brooks SA, Lear TL, Adelson DL, Bailey E.Tobiano is a white spotting pattern in horses caused by a dominant gene, Tobiano(TO). Here, we report TO associated with a large paracentric chromosome inversion on horse chromosome 3. DNA sequences flanking the inversion were identified and a PCR test was developed to detect the inversion. The inversion was only found in horses with the tobiano pattern, including horses with diverse genetic backgrounds, which indicated a common genetic origin thousands of years ago. The inversion does not interrupt any annotated genes, but begins approximately 100 kb downstream of the KIT gene. This inversion...
Theegarten D, Sachse K, Mentrup B, Fey K, Hotzel H, Anhenn O.Recurrent airway obstruction (RAO) in horses is a naturally occurring dust-induced disease mainly characterized by bronchiolitis which shows histological and pathophysiological similarities to human chronic obstructive pulmonary disease (COPD). In human COPD previous investigations indicated an association with Chlamydophila psittaci infection. The present study was designed (1) to clarify a possible role of this infectious agent in RAO and (2) to investigate the suitability of this equine disorder as a model for human COPD. Methods: Clinico-pathological parameters of a total of 45 horses (25 ...
Traversa D, Fichi G, Campigli M, Rondolotti A, Iorio R, Proudman CJ, Pellegrini D, Perrucci S.Anoplocephala perfoliata (Cestoda, Cyclophyllidea), the commonest intestinal tapeworm of horses, can cause colic, intussusceptions, ileal impactions and intestinal perforations. Common diagnostic techniques for A. perfoliata infection, i.e. coprology and serology, show inherent limitations in terms of sensitivity and specificity and new approaches are thus required. Hence, the present study compared the reliability of coprological, serological (i.e. ELISA) and molecular (i.e. nested PCR) methods in detecting A. perfoliata infection in naturally infected horses and in horses treated with a comb...
Varcasia A, Garippa G, Pipia AP, Scala A, Brianti E, Giannetto S, Battelli G, Poglayen G, Micagni G.Between March 2003 and February 2007, the livers and the lungs of 2,231 horses from various Italian regions were examined for cystic echinococcosis presence at the time of slaughter. Hydatid cysts were found in six horses, namely four from Sardinia, one from Sicily, and one from Tuscany. The location, number, morphology, and fertility of the cysts found were determined. DNA was extracted from the germinal layers and protoscoleces of the fertile cysts and polymerase chain reactions (PCR) were performed in order to strain type DNA isolates for reduced nicotinamide adenine dinucleotide dehydrogen...
Bell SA, Pusterla N, Balasuriya UB, Mapes SM, Nyberg NL, MacLachlan NJ.Equids are commonly infected by herpesviruses, but isolation of herpesviruses from mules has apparently not been previously reported. Furthermore, the genomic relationships among the various equid herpesviruses are poorly characterized. We describe the isolation and preliminary characterization of a mule gammaherpesvirus tentatively identified as asinine herpesvirus-2 (AHV-2; also designated equid herpesvirus-7 (EHV-7)) from the nasal secretions (NS) of a healthy mule in northern California. The virus was initially identified by transmission electron microscopic examination of lysates of cell ...
Ball BA, Sabeur K, Allen WR.Sperm-mediated gene transfer has been reported as a method for production of transgenic animals in a variety of species, and this technique represents a possible method for production of transgenic equids. Objective: To evaluate the uptake of exogenous DNA (enhanced green fluorescent protein; pEGFP) by equine spermatozoa and to assess the ability of transfected spermatozoa to introduce this transgene into early equine embryos. Methods: To evaluate incorporation of pEGFP into equine spermatozoa, washed spermatozoa were incubated with 32P-pEGFP, with or without lipofection. Spermatozoa were also...
Visser MB, Pollitt CC.Laminitis is known to involve deregulation of proteases and destruction of the lamellar basement membrane with the host inflammatory response also playing a role. Leukocyte infiltration has been well characterized in the black walnut model of laminitis induction, but not in carbohydrate induced models. Increased gene expression of multiple cytokines, including IL-6, has also been implicated in laminitis development. Using real time PCR, immunohistochemistry and zymography methods, we characterize leukocyte infiltration and IL-6 gene expression in oligofructose (OF) induced laminitis. As well, ...
Galosi CM, Vila Roza MV, Oliva GA, Pecoraro MR, Echeverría MG, Corva S, Etcheverrigaray ME.Equine herpesvirus 1 (EHV-1) is the causative agent of abortion, perinatal foal mortality, neurological and acute respiratory diseases in horses. Conventional laboratory diagnosis involving viral isolation from aborted foetuses is laborious and lengthy and requires processing of samples within 24 h of collection, which is problematic for samples that come from long distances. The aim of this study was to develop a polymerase chain reaction (PCR) assay useful in Argentina to detect DNA sequences of EHV-1 in different tissues from aborted equine foetuses with variable quality of preservation and...
Baldani CD, Nakaghi AC, Machado RZ.Blood and serum samples from 170 horses raised in the Jaboticabal microregion, São Paulo State, Brazil, were collected and tested by microscopic examination of blood smears, indirect fluorescent antibody test (IFAT) and nested polymerase chain reaction (nPCR) for Theileria equi infections. The association among the test results was verified by the McNemar test. During the examination of thin blood smears, parasites were detected in six (3.52%) horses. Anti-T. equi antibodies were detected in 100% sera samples, with titers ranging between 1:80 and 1:5120. The nPCR based on the T. equi merozoit...
Tarr CJ, Thompson PN, Guthrie AJ, Harper CK.The carrier prevalence of severe combined immunodeficiency (SCID), lavender foal syndrome (LFS) and cerebellar abiotrophy (CA) in Arabian foals in South Africa was determined in order to quantify the potential impact of these conditions locally. Furthermore, the carrier prevalence of SCID prior to and following the introduction of a genetic test was compared to evaluate the effect of testing in the population. Objective: To estimate the carrier prevalence of SCID, LFS and CA in registered purebred Arabians born in South Africa in the 2004/5 and 2009/10 foaling seasons and compare the changes i...
Szober CM, Hauck SM, Euler KN, Fröhlich KJ, Alge-Priglinger C, Ueffing M, Deeg CA.The purpose of this study was to characterize the cell surface proteome of native compared to cultured equine retinal pigment epithelium (RPE) cells. The RPE plays an essential role in visual function and represents the outer blood-retinal barrier. We are investigating immunopathomechanisms of equine recurrent uveitis, an autoimmune inflammatory disease in horses leading to breakdown of the outer blood-retinal barrier and influx of autoreactive T-cells into affected horses' vitrei. Cell surface proteins of native and cultured RPE cells from eye-healthy horses were captured by biotinylation, an...
Lee DH, Lee EB, Seo JP, Ko EJ.Despite vaccination, equine influenza virus (EIV) and equine herpesvirus (EHV) infections still cause highly contagious respiratory diseases in horses. Recently, concurrent vaccination with EIV and EHV was suggested as a new approach; however, there have been no reports of concurrent vaccination with recombinant canarypox EIV and inactivated EHV vaccines. In this study, we aimed to compare the EIV-specific immune responses induced by concurrent administrations of a recombinant canarypox EIV vaccine and an inactivated bivalent EHV vaccine with those induced by a single recombinant canarypox EIV...
Szeredi L, Hornyák A, Pálfi V, Molnár T, Glávits R, Dénes B.The occurrence of equine arteritis virus (EAV) induced equine abortions was studied with different laboratory methods during a 3-year period. Tissue samples from 96 aborted equine foetuses or newborn foals were collected from 57 farms located in different parts of Hungary. Virus isolation, polymerase chain reaction (PCR), immunohistochemistry and serology were used for the detection of EAV infection. The overall seroprevalence of EAV infection in mares was 65%. EAV induced abortion was diagnosed in eight (8.3%) cases from six (10.5%) herds. Abortion was sporadic in all herds except for one, wh...
Pavone S, Sforna M, Gialletti R, Prato S, Marenzoni ML, Mandara MT.A 7-year-old male trotter horse with a history of recurrent colic displayed clinical findings consistent with chronic intestinal pseudo-obstruction (CIP). At laparotomy, an impaction of the descending colon associated with marked atrophy of the right dorsal colon was found. The horse was humanely destroyed and tissues collected at necropsy examination revealed diffuse enteric ganglionitis comprising an infiltrate of CD3(+) T lymphocytes and plasma cells. At all levels of the intestinal tract the number of myenteric ganglia and of normal ganglion cells was decreased significantly. There were ch...
Hassanien RT, Hussein HA, Abdelmegeed HK, Abdelwahed DA, Khattab OM, Ali MH, Habashi AR, Ibraheem EM, Shahein MA, Abohatab EM.Due to climatic changes, arthropod-borne viruses have become a global health concern. In Egypt, West Nile virus (WNV) was initially detected in humans in 1950 and then in 1951, 1954, 1968, and 1989. Although WNV infection has been recorded in numerous Middle Eastern countries, its prevalence among the equine population in Egypt is unknown. This study aimed to investigate the current situation of vector-borne WNV in Egypt, estimate its seroprevalence, and assess the associated risk factors. Unassigned: We screened 1100 sera samples and nasal swabs from the same equids, 156 mosquito pools, and 3...
Madrigal-Valencia TL, Saavedra-Montañez M, Pérez-Torres A, Hernández J, Segalés J, Hernández YD, Candanosa-Aranda IE, Pérez-Guiot A....Ovine gammaherpesvirus 2 (OvHV-2), a member of the genus Macavirus, causes sheep-associated malignant catarrhal fever (SA-MCF), a fatal lymphoproliferative disease affecting a wide variety of ungulates in addition to horses. This study described an outbreak of SA-MCF in Mexico and the identification of the OvHV-2 virus in primary rabbit testis cultures through the generation of intranuclear inclusion bodies, syncytia, immunofluorescence (IF), immunocytochemistry (ICC), immunohistochemistry (IHC), endpoint polymerase chain reaction (PCR), and partial sequencing of the ORF75 gene. The animals in...
Wilsher S, Omar H, Ismer A, Allen T, Wernery U, Joseph M, Mawhinney I, Florea L, Thurston L, Duquesne F, Petry S.Three horse mares inadvertently inseminated with semen from a Tayorella asinigenitalis-positive Jack donkey developed severe, purulent endometritis whereas two Jenny donkeys mated naturally to the same Jack donkey did not develop clinical signs of infection. Objective: To isolate and identify the causative agent. Methods: Case report. Methods: Endometrial swabs from the infected mares were cultured on selective and non-selective media under aerobic and microaerophilic conditions. Isolates were subjected to Gram staining, oxidase and catalase tests, the Monotayl Latex Agglutination test and PCR...
Venner M, Heyers P, Strutzberg-Minder K, Lorenz N, Verspohl J, Klug E.The goal of the present study was to investigate whether new PCR-methods would improve diagnostic of R. equi. In a first step, sensitivity and specificity of the PCR-methods in respect to the"gold standard" microbiological culture were determined. Secondly, sensitivity and specificity of both microbiological methods were evaluated in respect to the clinical diagnosis. The tracheobronchial secretions of 48 foals with pulmonary abscesses and of 37 healthy foals were evaluated by bacteriological culture as well as by four PCR-methods: aceA-, ideR-, vapA- and VP-PCR. In respect to the"gold standar...
Rizvi SM, Slater JD, Wolfinger U, Borchers K, Field HJ, Slade AJ.The distribution of equine herpesvirus 2 (EHV-2) DNA within neurological and lymphoid tissues from 12 EHV-2 seropositive Welsh mountain ponies was determined by PCR. The lymphoid sites sampled in this study were almost universally PCR positive, thus confirming the existing virus co-cultivation data which suggest that the lymph nodes draining the respiratory tract are the main reservoirs of EHV-2 DNA. In addition, EHV-2 DNA was also detected, albeit with lower frequency, within both the peripheral and central nervous systems (PNS and CNS) of these animals. Of the CNS sites sampled 11% were PCR-...
Sharav T, Konnai S, Ochirkhuu N, Ts EO, Mekata H, Sakoda Y, Umemura T, Murata S, Chultemdorj T, Ohashi K.The genetic characterization and actual prevalence of EIAV in Mongolian horse in the disease endemic region is currently unknown. Here, 11 of 776 horse serum samples from four Mongolian provinces tested positive on agar gel immunodiffusion test. Genomic DNA extracted from all seropositive samples was subjected to nested PCR assay. Among these, three samples tested positive with nested PCR assay and were identified by sequencing analysis based on long termination repeat and tat gene of the virus. Two of the three sequences were identical, with 94.0% identity with the third. These two independen...
Norton EM, Wooldridge AA, Stewart AJ, Cusimano L, Schwartz DD, Johnson CM, Boudreaux MK, Christopherson PW.Hemophilia A is an X-chromosome-linked disorder caused by a deficiency in factor VIII (FVIII). Although foals have been diagnosed with hemophilia A based on deficiency in FVIII activity, causative gene mutations have not been identified. The genomic DNA and cDNA encoding FVIII of a Tennesee Walking Horse colt affected with hemophilia A and the genomic DNA of his dam and a normal unrelated horse were analyzed with no splice site or coding sequence abnormalities identified in any of the horses. Polymerase chain reactions (PCR) were then performed on hepatic cDNA from the affected colt and an unr...
Ekiri AB, Long MT, Hernandez JA.The main objective of this study was to assess the diagnostic performance of a real-time polymerase chain reaction (PCR) assay for the detection of Salmonella in fecal samples collected from hospitalized horses with or without signs of gastrointestinal (GI) tract disease. The PCR assay used primers and a probe that targeted the invA gene of Salmonella. Assuming a sensitivity of 100% and a specificity of 96.6%, and a disease prevalence of 2%, 5%, and 10-15% in study horses, the PCR assay had a high (100%) negative predictive value, and a positive predictive value that ranged from 37% in horses ...
Rahmani SS, Malekifard F, Tavassoli M.Neospora caninum is an Apicomplexan parasite that can cause enormous economic losses due to abortions in cattle. The present study investigated the role of Neospora spp. infection in equine abortion in Iranian donkeys using molecular and phylogenetic analyses. Twenty-nine-aborted fetuses and 29 blood samples from their dams were collected from six different regions in the West and Northwest of Iran. They were tested for N. caninum by PCR at the Nc5 locus, followed by sequencing of five of the PCR products. The overall molecular prevalence was 34.5% in blood samples and the prevalence by DNA de...
Silva FS, Lorenzett MP, Bianchi MV, Bastos HBA, Larentis GR, Paul LG, Snel GGM, Oliveira-Filho JP, Mattos RC, Sonne L.Although relatively uncommon in horses, infections caused by Mycobacterium spp. may affect the gastrointestinal tract. Mycobacterium branderi is a non-tuberculous Mycobacterium (NTM) that causes respiratory infections in man. Non-tuberculous mycobacteria may also affect horses; however, infection by M. branderi has not yet been reported in this species. This report describes the clinical, pathological, microbiological and molecular findings of M. branderi infection in a horse, causing granulomatous mesenteric lymphadenitis. A 17-year-old Thoroughbred stallion had a 3-month history of chronic d...
Aradaib IE.A nested reverse transcriptase (RT) polymerase chain reaction (RT-PCR), for rapid detection of African horse sickness virus (AHSV) double-stranded ribonucleic acid (dsRNA) in cell culture and tissue samples, was developed and evaluated. Using an outer pair of primers (P1 and P2), selected from genome segment three of AHSV serotype 6 (AHSV-6), the RT-PCR-based assay resulted in amplification of a 890 base pair (bp) primary PCR product. RNAs from the nine vaccine strains of AHSV, and a number of AHSV field isolates including the Central African isolates of AHSV-9 and AHSV-6, propagated in cell c...
Begg AP, Carrick J, Chicken C, Blishen A, Todhunter K, Eamens K, Jenkins C.This report describes the fetoplacental pathology of associated abortion, premature birth, and neonatal loss in 46 of 442 equine abortion investigations between 2015 and 2019. Seven abortions, 26 premature births, and 13 neonatal deaths with positive polymerase chain reaction (PCR) were evaluated. In 83% of cases (38/46), infection was considered as the primary cause of loss based on quantitative PCR (qPCR) confirmation, pathological findings, and exclusion of other causes, and was supported by immunolabeling in fetoplacental lesions. Lymphohistiocytic placentitis with vasculitis (36/38) af...
Di Bernardo G, Del Gaudio S, Galderisi U, Cipollaro M.Ancient DNA extracted from 2000 year-old equine bones was examined in order to amplify mitochondrial and nuclear DNA fragments. A specific equine satellite-type sequence representing 3.7%-11% of the entire equine genome, proved to be a suitable target to address the question of the presence of aDNA in ancient bones. The PCR strategy designed to investigate this specific target also allowed us to calculate the molecular weight of amplifiable DNA fragments. Sequencing of a 370 bp DNA fragment of mitochondrial control region allowed the comparison of ancient DNA sequences with those of modern hor...
Hensel M, Meason-Smith C, Plumlee QD, Myers AN, Coleman MC, Lawhon S, Rodrigues Hoffmann A, Rech RR.Pulmonary mycosis secondary to enterocolitis is an uncommon diagnosis in equine medicine, but is thought to result from mucosal compromise and translocation of enteric fungi. The aetiological agent associated with translocation is often identified based on fungal culture or hyphal features in histological sections. In order to understand better the aetiological agents involved, six horses diagnosed with Salmonella enteritis and concurrent pulmonary mycosis were identified retrospectively through a database search of veterinary teaching hospital records. Samples from these cases were subjected ...
Alinovi CA, Ward MP, Couëtil LL, Wu CC.To assess methods of detecting environmental contamination with Salmonella organisms and evaluate a cleaning and disinfection protocol for horse stalls in a veterinary teaching hospital. Methods: Original study. Methods: 37 horses with diarrhea likely to be caused by Salmonella infection and their stall environments. Methods: Fecal samples were collected from horses daily during hospitalization; samples were obtained from stall sites after cleaning and application of disinfectants. Fecal and environmental samples were cultured for Salmonella spp and tested via polymerase chain reaction (PCR) a...
Díaz S, Giovambattista G, Peral-García P.Sequence information was obtained on the variation of the ELA-DRB upstream regulatory region (URR) after polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) cloning and sequencing of approximately 220 bp upstream of the first exon of horse DRB genes. The sequence of the proximal URR of equine DRB is composed of highly conserved sequence motifs, showing the presence of the W, X, Y, CAAT and TATA conserved boxes of major histocompatibility complex (MHC) class II promoters. Five different polymorphic horse DRB promoter sequences were detected in five horse breeds. The res...
Andrews ES, Turell MJ.Emerging and re-emerging arboviruses continue to be a threat to global public health, and viral surveillance of mosquito populations is critical for mosquito control operations. Due to the tropical climate of many of the affected areas, it may be difficult to maintain a cold chain as the samples travel from collection sites to laboratories for testing. We determined how suboptimal holding temperatures affected the ability to detect viruses in pools of mosquitoes. Adult female Aedes albopictus and Ae. taeniorhynchus individuals were inoculated with chikungunya virus or Venezuelan equine encepha...
Dzieciatkowski T, Przybylski M, Cymerys J, Turowska A, Chmielewska A, Tucholska A, Banbura MW.Equine herpesvirus type 1 (EHV-1) is one of the major viral agents causing diseases in horses common worldwide. A variety of techniques, including PCR, have been used to diagnose EHV-1 infections. In this paper, an attempt of real-time PCR has been described, which uses specific fluorochrome-labeled TaqMan probes for detection of viral DNA. This method does not require post-amplification manipulations, thereby reducing the risk of cross-contamination. The assay was sensitive enough to detect EHV-1 sequences in different clinical samples, as well in mice neuronal cell cultures. The technique wa...
Takai S, Vigo G, Ikushima H, Higuchi T, Hagiwara S, Hashikura S, Sasaki Y, Tsubaki S, Anzai T, Kamada M.Polymerase chain reaction (PCR)-based assays were developed to detect virulent Rhodococcus equi in transtracheal aspirate samples from sick foals showing respiratory signs. An oligonucleotide primer pair from the sequence of the virulence-associated 15- to 17-kDa antigen gene of the virulence plasmid in virulent R. equi was used to amplify a 564 bp region by PCR, and the result was confirmed by Southern blot hybridization. No positive reaction was seen in DNA from 13 different microorganisms typically found in the respiratory tract. In tracheal aspirates seeded with virulent R. equi, a visible...
Koekemoer JJ, Dijk AA.Since protection against African horsesickness (AHS) is serotype-specific, rapid serotyping of AHSV is crucial to identify the correct vaccine serotype for efficient control of the spread of AHS outbreaks, especially when they occur in non-endemic regions. This paper describes the first one-day serotyping procedure that requires only a single RT-PCR and hybridization and which can identify multiple serotypes in mixed infections in one assay. The same region of genome segment 2 of all nine AHSV serotypes is amplified in a single RT-PCR. A universal primer set, designed to amplify the 5'-termina...
Tomczuk K, Hirzmann J, Köhler K, Szczepaniak K, Studzinska M, Demkowska-Kutrzepa M, Roczeń-Karczmarz M, Bauer C.A cross-sectional survey was performed to obtain first information on the prevalence of Echinococcus multilocularis infection in slaughter horses from central, eastern and southern Poland, a country with a highly endemic occurrence of this tapeworm in red foxes. White tough nodular lesions being 3-10 mm in size, sharply demarcated and spherically or irregularly shaped were found in 54 of 365 livers examined. Histologically, focal granulomatous necrotizing inflammations and sometimes PAS-positive acellular lamellar structures being characteristic of the E. multilocularis metacestode stage were...
Mahlobo SI, Zishiri OT.Ticks are medically important and significant vectors of diseases affecting livestock, humans, and companion animals than any other arthropod vectors. In the absence of information on the relationship of tick species and piroplasms parasites in Lesotho, the current study was aimed at detecting piroplasms parasites of economic importance from ticks of domestic animals. A total of 322 pooled tick DNA samples were subjected to PCR screening for the presence of piroplasms. The overall infection rate of piroplasms was 7% with Babesia bigemina at 3.4% (11/322), B. bovis 0.3% (1/322), B. ovis 2.8% (9...
