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.
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 ...
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....
Abdul-Latif SAK, Yousif AA.Strangles is a highly contagious equine respiratory disease caused by . It is a globally significant pathogen and one of the most common infectious agents in horses. In Iraq, no sequencing data on this pathogen are available, and only two molecular studies have been published to date. This study provides preliminary insights into strain diversity and provides a foundation for future large-scale investigations. Unassigned: This study aimed to investigate the molecular characteristics, identify gene alleles, and perform a phylogenetic analysis of isolates from horses in Baghdad, Iraq. Unassign...
Stefaniuk-Szmukier M, Bieniek A, Ropka-Molik K, Bellone RR.Congenital stationary night blindness (CSNB) has been connected to the leopard complex spotting phenotype (LP) in various horse breeds. CSNB associated with LP is thought to be caused by a 1378 bp insertion in TRPM1, with homozygotes being nightblind and having few to no spots of pigment in their white patterned area. This study aimed to assess the prevalence of CSNB alleles in tarant-colored horses in Poland through a three-primer system for an allele-specific Polymerase Chain Reaction (PCR). The TRPM1 gene insertion was genotyped in 221 horses belonging to Małopolska, Felin and Shetland Pon...
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 ...
Diekmann I, Krücken J, Kuzmina TA, Bredtmann CM, Louro M, Kharchenko VA, Tzelos T, Matthews JB, Madeira de Carvalho LM, von Samson-Himmelstjerna G.Morphologically, 64 strongylid species have been described in equines. Co-infections are common, with up to 29 species reported in a single horse. Morphological identification of these species is time consuming and requires expert knowledge due to their similar appearance. Therefore, non-invasive identification methods are needed. DNA barcoding offers a rapid and reliable tool for species identification and the discovery of cryptic species for these most common parasitic nematodes of equines. In total, 269 COI and 312 ITS-2 sequences from 27 equine Strongylidae species, including sequences fro...
Petano-Duque JM, Urueña-Martinez E, Cabezas-Callejas LL, Perilla-Amaya J, Rueda-García V, Rondón-Barragán IS, Lopera-Vásquez R.EHV-1 is one of the most prevalent viral pathogens in horses; however, its prevalence is not well defined by cross-reactions of serological tests. Thus, this study aims to determine EHV-1/-4 prevalence in Colombian creole horses from Ibagué, Tolima, using molecular techniques, as well as to determine possible risk factors associated with viral infection. A cross-sectional study was carried out in 110 equines blood, serum, and semen samples from Ibagué, Tolima. Antibodies against EHV-1/-4 were determined through indirect ELISA. EHV-1 was detected by amplifying the glycoprotein H gene through ...
Pusterla N, Barnum S, Lawton K, Craig B, James K.Equid alphaherpesvirus 4 (EqAHV4; Orthoherpesviridae, Varicellovirus equidalpha4; equine rhinopneumonitis virus) has seldom been associated with complications such as abortion and myeloencephalopathy, given the low tendency of this virus to induce viremia. We investigated the frequency of EqAHV4 viremia in horses with fever and respiratory signs. Case selection included all equids with EqAHV4 quantitative real-time PCR (qPCR)-positive nasal secretions (defined as EqAHV4 qPCR-positive cases) submitted to a diagnostic laboratory. Controls consisted of each case submitted before and after each Eq...
Kabir A, Kelley WG, Glover C, Erol E, Helmy YA.Salmonella is a foodborne pathogen that poses a significant threat to global public health. It affects several animal species, including horses. Salmonella infections in horses can be either asymptomatic or cause severe clinical illness. Infections caused by Salmonella are presently controlled with antibiotics. Due to the formation of biofilms and the emergence of antimicrobial resistance, the treatment has become more complicated. Our study focused on investigating the prevalence of Salmonella enterica in necropsied horses, assessing the capability for biofilm formation, and motility, determi...
Nemoto M, Kawanishi N, Kambayashi Y, Bannai H, Yamanaka T, Tsujimura K.Equine influenza virus (EIV) can be transmitted by inhalation of aerosolized droplets, direct contact, and contaminated fomites. However, to our knowledge, there are no reports of the recovery of EIV from the air surrounding infected horses. Here, we evaluated whether EIV can be recovered from the air in the stalls of experimentally infected horses by using an air sampler. Furthermore, we examined whether rapid molecular test kits with reaction times of less than 30 min can detect EIV from air samples for potential field application. Two horses kept in individual stalls were experimentally i...
Sharma NK, Singh P, Saha B, Bhardwaj A, Iquebal MA, Pal Y, Nayan V, Jaiswal S, Giri SK, Legha RA, Bhattacharya TK, Kumar D, Rai A.Copy number variations (CNVs) have become widely acknowledged as a significant source of genomic variability and phenotypic variance. To understand the genetic variants in horses, CNVs from six Indian horse breeds, namely, Manipuri, Zanskari, Bhutia, Spiti, Kathiawari and Marwari were discovered using Axiom™ Equine Genotyping Array. These breeds differed in agro-climatic adaptation with distinct phenotypic characters. A total of 2668 autosomal CNVs and 381 CNV regions (CNVRs) were identified with PennCNV tool. DeepCNV was employed to re-validate to get 883 autosomal CNVs, of which 9.06% were...
Cabete A, Xufre Â, Padre L, Bettencourt E, Nunes T, Gomes J.Equine piroplasmosis (EP) is a tick-borne disease of equids caused by Theileria equi, Theileria haneyi, and Babesia caballi. EP is endemic in most tropical and subtropical regions worldwide, and there is a likelihood that it is also endemic in Portugal. This retrospective study aimed to determine the seroprevalence, prevalence, and potential risk factors of EP in our country over the past five years. A total of 3063 diagnostic test records were analysed. Results from the competitive enzyme-linked immunosorbent assay (cELISA) revealed a seroprevalence of 32.7 % and 15.7 % for T. equi and B....
Nassar AFC, Chiebao DP, Fava CD, Miyashiro S, Castro V, Ogata RA, Yamamora JM, Monteiro CAS, Monteiro EJB.Glanders is a zoonotic disease of equids caused by the bacterium Burkholderia mallei, responsible for considerable economic loss. This study aimed to describe the clinical manifestations, pathological findings, and also bacteriological and molecular methods for agent detection in naturally infected animals (16 adult horses and one fetus) detected by serological survey from three glanders outbreaks. Of the 16 horses, 6 (37.5%) did not show clinical signs. After necropsy,samples of organs, lymph nodes, lesions and secretions were collectedfor histopathology, bacterial isolation, and PCR. The cli...
Axt CW, Springer A, von Luckner J, Naucke TJ, Müller E, Strube C, Schäfer I.Equine piroplasmosis (EP) is caused by (.) and (.) and is transmitted by hard ticks. Predominantly, the Mediterranean region is known as being endemic for both pathogens in Europe. However, autochthonous infections in central European countries such as Germany can no longer be ruled out due to individual case reports in horses without any stays abroad as well as the geographical expansion of the habitats of different tick species. The case reports presented underline the risk of infection for horses travelling to endemic areas and in horses imported from such areas to non-endemic countries...
Solbach V, Grabatin M, Zablotski Y, Fux R, Zerbe H, Witte TS.Contagious Equine Metritis (CEM) caused by the bacterium Taylorella equigenitalis (T. equigenitalis), is a venereal infection of equids which is of international concern to the equine breeding industry. A recent study showed a high prevalence of T. equigenitalis in Icelandic stallions when compared to stallions of other breeds also using for natural breeding. Consequently, the objectives of the present study were to investigate the prevalence of T. equigenitalis in Icelandic mares and geldings and to determine factors associated with a T. equigenitalis-positive qPCR result. In total, 361 Icela...
Charles A, Kerckhove HV, De Maré L, Cassart D, Ficheroulle J, Pouyade GR, Tosi I.A Standardbred racehorse was presented for exercise intolerance, weight loss, pyrexia and facial deformity. Radiography and ultrasonography revealed periostitis and regional soft tissue swelling of maxillary bones. Computed tomography excluded any dental or sinus origin of these abnormalities. Further deformities on distal limbs and skin lesions appeared during hospitalization. Radiography identified bilateral periostitis and soft tissue swelling in the distal radius and metatarsal bones, as observed in the head, suggestive of hypertrophic osteopathy (HO). Skin biopsies revealed granulomatous ...
Paulino PG, Amaral FB, de Oliveira RT, de Andrade SG, Rabello CA, Meirelles N, de Souza Santana M, Galdino KCP, Jacob JCF, Peckle M, Massard CL....Equine piroplasmosis, a tick-borne disease caused by hemoparasites of the Babesia and Theileria genera, has significant clinical and economic impacts worldwide. This study aims to characterize the heat shock protein 70 kDa (hsp70) gene sequences of Theileria equi from naturally infected horses across the five geographic regions in Brazil, and to analyze the phylogenetic relationships between T. equi and other parasites within the order Piroplasmida. Primers specific to T. equi were designed through in silico analysis of hsp70 gene sequences available in GenBank. Forty samples of equine whole ...
Phetkarl T, Fungwithaya P, Lewchalermvong K, Sontigun N.The global prevalence of gastrointestinal (GI) and blood parasite infections in horses is a significant concern due to their substantial impact on morbidity, mortality, and economic losses in the horse industry. In Thailand, limited research has been conducted on these parasites in horse populations, and data from southern Thailand are lacking. Consequently, this study aimed to estimate the prevalence of GI and blood parasites in horses in Nakhon Si Thammarat province, Thailand. Unassigned: In total, 79 fecal and blood samples were collected from horses across 11 farms in Nakhon Si Thammarat p...
Parkinson NJ, Ward A, Malbon AJ, Reardon RJM, Kelly PG.Equine sarcoids are common non-metastasising skin tumours in horses, associated with bovine papillomavirus (BPV) infection. Six subtypes are recognised (occult, verrucose, nodular, fibroblastic, mixed and malevolent lesions), with variable clinical behaviour. The pathophysiology underlying varying tumour phenotype is poorly understood, and previous data on associations with viral load have been conflicting. To better understand this clinical variation, we investigated associations between tumour subtype and viral load, viral early protein gene expression, and expression of 10 host genes by qua...
de Oliveira UV, Varjão JL, de Jesus Deiró AG, Maciel BM, Silva FL, Pinheiro AM, Gondim LFP, Munhoz AD.The aim of this study was to isolate from equids destined for slaughter in a Brazilian slaughterhouse. A total of 354 equids were analyzed, with blood samples collected from all the animals and samples of masseter muscle and brain tissue collected from 319 animals. A serological test was conducted to detect equids with specific antibodies for . Molecular detection of by nested PCR was performed on the tissue samples collected. Tissue samples were tested by murine bioassay in an attempt to isolate either the parasite or the parasite DNA. Real-time PCR was performed on the brain samples from 1...
Cornell TR, Fye BL, Nyassi E, Ceesay F, Jallow M, Langendonk RF, Wootton DG, Pinchbeck G, Scantlebury CE.Exposure rates to species, the causative agent of equine epizootic lymphangitis (EL), are unknown amongst working equids in The Gambia. The primary aims of this study were to estimate anti- antibody seroprevalence in the equid population in rural The Gambia and to explore risk factors for seropositivity. Unassigned: A nationwide cross-sectional study was conducted (February-July 2022), representing baseline measurements of a longitudinal cohort study. Horses ( 463) and donkeys ( = 92) without EL signs were recruited in 18 study sites. Following informed owner consent, equid clinical and m...
Lawrence KE, Gedye K, Carvalho L, Wang B, Fermin LM, Pomroy WE.To determine whether evidence for infection with (Ikeda) could be identified in samples of commercial red deer , horses, and working farm dogs in New Zealand. Unassigned: Blood samples were collected during October and November 2019 from a convenience sample of red deer (n = 57) at slaughter. Equine blood samples (n = 50) were convenience-sampled from those submitted to a veterinary pathology laboratory for routine testing in January 2020. Blood samples, collected for a previous study from a convenience sample of Huntaway dogs (n = 115) from rural regions throughout the North and ...
Ramsay L, Eberhardt C, Schoster A.Reports of leptospirosis in horses are limited. Objective: To describe the clinical and diagnostic findings of acute systemic leptospirosis in horses. Methods: Eleven client-owned horses presented to an equine hospital because of acute onset of disease between 2015 and 2023. Methods: Retrospective case series. Horses diagnosed with leptospirosis by 1 or more of urine PCR, serologic microscopic agglutination test (MAT), and histopathology. Results: Common clinical signs included lethargy (10), anorexia (10), fever (9), tachypnea (9), abnormal lung sounds (9), and epistaxis (6). Acute kidney inj...
Bekele D, Dessalegn B, Tadesse B, Abey SL.Strangles, caused by Streptococcus equi subspecies equi, is a highly infectious disease of equines causing major health issues and financial losses. The aim of the study was to detect the presence of the SeM gene in Streptococcus equi isolated from equine suspected of having strangles. A cross-sectional study design was conducted from July to December 2022 in five districts of the central Gondar zone, Ethiopia. One-hundred sixty swab samples were taken from animals that had been clinically suspected. The SeM gene was detected using polymerase chain reaction, and the antimicrobial susceptibilit...
Mendoza FJ, Pérez-Écija A, Kappmeyer LS, Suarez CE, Bastos RG.Equine piroplasmosis (EP) is a global tick-borne disease of equids caused by the intraerythrocytic apicomplexan parasites and , and the more recently discovered . These parasites can be transmitted by several tick species, including , , and , but iatrogenic and vertical transmission are also common. Clinical signs of EP include poor performance, fever, icterus, abortions, among others, and peracute or acute forms of infection are associated with high mortality in non-endemic areas. EP is a reportable disease and represents an important barrier for the international trade of horses and other e...
Bermukhametov Z, Suleimanova K, Tomaruk O, Baimenov B, Shevchenko P, Batyrbekov A, Mikniene Z, Onur Girişgin A, Rychshanova R.A total of 396 samples were taken from the hearts, oesophagi, and diaphragms of 132 horses slaughtered at slaughterhouses in 2023 for subsequent examination. Methods: The histological method revealed pathomorphological changes in the muscle tissue. The molecular method identified the pathogen species. Results: Histological examination revealed thick-walled cysts with internal septa and numerous bradyzoites, and mononuclear inflammatory cells with pericyst infiltrates. Microcyst samples were amplified by polymerase chain reaction. Molecular genetic analysis allowed for the identification of 18 ...
Martínez-Sáez L, Pala S, Marín-García PJ, Llobat L.Toxoplasmosis is one of the most common parasitic zoonoses and represents a significant health risk for humans, especially for immunodeficient patients. The main transmission route is by oral uptake of oocysts and consumption of undercooked meat of infected animals. Different species have been evaluated as possible reservoirs of the parasite, but few studies have been carried out to examine the role of horses in transmission of the disease. Given the proximity of these animals to humans and the widespread consumption of their meat in many countries, including the Mediterranean basin, it is imp...
Stasiak K, Dunowska M, Rola J.Equine rhinitis A (ERAV) and B (ERBV) viruses are respiratory pathogens with worldwide distribution. The current study aimed to determine the frequency of infection of ERAV and ERBV among horses and foals at Polish national studs, and to determine genetic variability within the viruses obtained. Virus-specific quantitative RT-PCR assays targeting a 5' untranslated region were used to screen nasal swabs collected from 621 horses at 16 national horse studs from throughout Poland, including 553 healthy horses and 68 horses with respiratory disease. A partial DNA polymerase gene was amplified and ...
Lacante SA, Jiang C, Mustamir AA, Mizuno T, Bi X, Syafruddin D, Tokoro M., a human intestinal protozoan parasite of the diplomonad group, has been overlooked because of its commensal features; therefore, molecular studies on this parasite are limited. To address this gap, we designed a molecular screening protocol using polymerase chain reaction (PCR) and DNA sequencing targeting the 18S small subunit ribosomal RNA gene and applied this screening method to the molecular epidemiological analysis of spp. in humans and various livestock. We validated our methodology using stool samples collected from 215 humans and 270 animal hosts (buffaloes, pigs, dogs, goats, hors...
Rar V, Marchenko V, Suntsova O, Epikhina T, Tikunov A, Meltsov I, Fedorets V, Igolkina Y, Kozlova I, Tikunova N.Equine piroplasmosis (EP) is a global worldwide infection, which can lead to the death of animals. Despite the causative agents of EP being well studied, there are no data on the distribution and genetic characteristics of EP agents in any region of Russia. In this study, blood samples from 750 horses from Novosibirsk province, Irkutsk province, and Altai region of Russian Siberia were examined for the presence of EP agents. Theileria equi and Babesia caballi were detected in all examined regions, with mean prevalence rates of 60.4% and 7.2%, respectively. The identified pathogens were genetic...
Hamond C, Martins G, Loureiro AP, Pestana C, Lawson-Ferreira R, Medeiros MA, Lilenbaum W.The aim of the present study was to consider the wide usage of urinary PCR as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock. A total of 512 adult animals (300 cattle, 138 horses, 59 goats and 15 pigs), from herds/flocks with reproductive problems in Rio de Janeiro, Brazil was studied by serology and urinary PCR. From the 512 serum samples tested, 223 (43.5 %) were seroreactive (cattle: 45.6 %, horses: 41.3 %, goats: 34%and pigs: 60 %). PCR detected leptospiral DNA in 32.4 % (cattle: 21.6 %, horses: 36.2 %, goats: 77.4 % and pigs: 33.3 %. To our knowledge t...
Williams KJ, Maes R, Del Piero F, Lim A, Wise A, Bolin DC, Caswell J, Jackson C, Robinson NE, Derksen F, Scott MA, Uhal BD, Li X, Youssef SA, Bolin SR.Pulmonary fibrosis and interstitial lung disease are poorly understood in horses; the causes of such conditions are rarely identified. Equine herpesvirus 5 (EHV-5) is a gamma-herpesvirus of horses that has not been associated with disease in horses. Pathologic and virologic findings from 24 horses with progressive nodular fibrotic lung disease associated with EHV-5 infection are described and compared with 23 age-matched control animals. Gross lesions consisted of multiple nodules of fibrosis throughout the lungs. Histologically, there was marked interstitial fibrosis, often with preservation ...
Hijjawi N, Mukbel R, Yang R, Ryan U.Little is known about the epidemiology of Cryptosporidium in Jordan and to date, only one genotyping study has been conducted on Cryptosporidium isolates from Jordanian children. In the present study, a total of 284 faecal samples from Jordanian cattle, sheep, goats and chicken and 48 human faecal samples were screened for the presence of Cryptosporidium using an 18S quantitative PCR (qPCR) and a C. parvum/C. hominis specific qPCR at a lectin locus. Of these, 37 of 284 animal faecal samples were positive by qPCR at the 18S locus giving an overall prevalence of 11.6%. The point prevalence of Cr...
Loncaric I, Künzel F, Licka T, Simhofer H, Spergser J, Rosengarten R.The aim of this study was to investigate the antimicrobial resistance, resistance gene patterns and genetic relatedness of a collection of Austrian methicillin-resistant Staphylococcus aureus (MRSA) isolates from companion animals and horses. A total of 89 non-repetitive MRSA isolates collected during routine veterinary microbiological examinations from April 2004 to the end of 2012, and one isolate from 2013 were used for this study. The presence of mecA and other resistance genes was confirmed by PCR. Isolates were genotyped by spa typing, two multiple-locus variable-number tandem repeat ana...
Johansson KE, Pettersson B, Uhlén M, Gunnarsson A, Malmqvist M, Olsson E.Seven Swedish isolates of Ehrlichia species from the blood of four dogs and three horses with clinical granulocytic ehrlichiosis, were identified by direct solid phase DNA sequencing of polymerase chain reaction (PCR) products from the 16S rRNA gene. The amplified DNA fragments were produced with primers complementary to the universal regions, U1, U2, U5 and U8 of the 16S rRNA molecule. Identical sequences were obtained from all seven isolates. This nucleotide sequence was similar to the sequences deposited in GenBank for Ehrlichia phagocytophila and E equi. The sequence of the Swedish ehrlich...
Torina A, Vicente J, Alongi A, Scimeca S, Turlá R, Nicosia S, Di Marco V, Caracappa S, de la Fuente J.The objective of this study was to characterize the observed prevalence of tick-borne pathogens (TBP) in domestic animals in Sicily, Italy during 2003-2005. Serological (competitive ELISA and indirect immunofluorescence antibody, n = 3299) and DNA tests (polymerase chain reaction and reverse line blot, n = 2565) were conducted on horse, donkey, cattle, sheep, goat, pig and dog samples. Pathogens analysed included Anaplasma, Ehrlichia, Rickettsia, Babesia and Theileria species, and Coxiella burnetii. The most prevalent TBP were Anaplasma and Babesia species. The results reported herein suggeste...
Perkins GA, Goodman LB, Tsujimura K, Van de Walle GR, Kim SG, Dubovi EJ, Osterrieder N.A single nucleotide polymorphism in the equine herpesvirus 1 (EHV-1) DNA polymerase gene (ORF30 A(2254) to G) has been associated with clinical signs of equine herpes myeloencephalopathy (EHM). The purpose of our study was to determine the odds ratio for this genetic marker and EHM using a panel of field isolates from North America collected over the past twenty-three years. EHV-1 isolates cultured at the Cornell University Animal Health Diagnostic Laboratory from 1984 to 2007 were retrieved along with their clinical histories. DNA was extracted from these EHV-1 cultures and allelic discrimina...
Angelini P, Tamba M, Finarelli AC, Bellini R, Albieri A, Bonilauri P, Cavrini F, Dottori M, Gaibani P, Martini E, Mattivi A, Pierro AM, Rugna G....Following a large West Nile virus (WNV) epidemic in northeastern Italy in 2008, human and animal surveillance activities were implemented in Emilia Romagna. Human surveillance was performed by serology or genome detection on blood and cerebrospinal fluid for all suspected cases suffering from acute meningoencephalitis in the regional territory. Animal surveillance consisted of passive and active surveillance of horses and active surveillance of wild birds and mosquitoes. Between 15 June and 31 October 2009, nine of 78 possible cases of West Nile neuroinvasive disease were confirmed (three fata...
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...
Laus F, Veronesi F, Passamonti F, Paggi E, Cerquetella M, Hyatt D, Tesei B, Fioretti DP.In order to investigate the prevalence of tick-borne diseases, equine piroplasmosis, equine granulocytic anaplasmosis and Lyme borreliosis in Central Italy, blood samples from 300 horses were analyzed for the presence of antibodies against Babesia caballi, Theileria equi, Anaplasma phagocytophilum and Borrelia burgdorferi using the IFAT. The blood samples were also subjected to PCR assays in order to detect pathogen DNA. A total of 78 (26.0%) and 123 (41.0%) horses were found to be seropositive for B. caballi and T. equi, respectively, while 41 (13. 4%) and 21 (7.0%) horses were, respectively,...
Marycz K, Weiss C, Śmieszek A, Kornicka K.Mesenchymal stem cells (MSCs) are frequently used in both human and veterinary medicine because their unique properties, such as modulating the immune response and differentiating into multiple lineages, make them a valuable tool in cell-based therapies. However, many studies have indicated the age-, lifestyle-, and disease-related deterioration of MSC regenerative characteristics. However, it still needs to be elucidated how the patient's health status affects the effectiveness of MSC differentiation. In the present study, we isolated mesenchymal stem cells from adipose tissue (adipose-derive...
Bogaert L, Van Poucke M, De Baere C, Peelman L, Gasthuys F, Martens A.Real-time quantitative PCR can be a very powerful and accurate technique to examine gene transcription patterns in different biological conditions. One of the critical steps in comparing transcription profiles is accurate normalisation. In most of the studies published on real-time PCR in horses, normalisation occurred against only one reference gene, usually GAPDH or ACTB, without validation of its expression stability. This might result in unreliable conclusions, because it has been demonstrated that the expression levels of so called "housekeeping genes" may vary considerably in different t...
Arroyo LG, Kruth SA, Willey BM, Staempfli HR, Low DE, Weese JS.Molecular typing of Clostridium difficile isolates from animals and humans may be useful for evaluation of the possibility for interspecies transmission. The objective of this study was to evaluate C. difficile isolates from domestic animals and humans using PCR ribotyping. Isolates were also tested using PCR for the presence of genes encoding toxins A and B. One hundred and thirty-three isolates of C. difficile from dogs (n = 92), horses (n = 21) and humans (n = 20), plus one each from a cat and a calf, were evaluated. Overall, 23 ribotypes were identified. Of these, nine were identified from...
Nikiforov TT, Rendle RB, Goelet P, Rogers YH, Kotewicz ML, Anderson S, Trainor GL, Knapp MR.A new method for typing single nucleotide polymorphisms in DNA is described. In this method, specific fragments of genomic DNA containing the polymorphic site(s) are first amplified by the polymerase chain reaction (PCR) using one regular and one phosphorothioate-modified primer. The double-stranded PCR product is rendered single-stranded by treatment with the enzyme T7 gene 6 exonuclease, and captured onto individual wells of a 96 well polystyrene plate by hybridization to an immobilized oligonucleotide primer. This primer is designed to hybridize to the single-stranded target DNA immediately...
Koehler K, Stechele M, Hetzel U, Domingo M, Schönian G, Zahner H, Burkhardt E.This report describes a case of cutaneous leishmaniosis in a horse in southern Germany. Diagnosis is based on histopathology, immunohistochemistry and electron microscopy. The protozoan was identified as Leishmania infantum via PCR and restriction fragment length polymorphism. The horse did not show specific Leishmania antibodies. The lesions healed completely within 6 months without any specific treatment. Since neither the infected horse nor its dam had ever left their rural area, autochthonous infection in Germany cannot be excluded. Factors possibly influencing the epidemiological situatio...
Sears WJ, Qvarnstrom Y, Dahlstrom E, Snook K, Kaluna L, Baláž V, Feckova B, Šlapeta J, Modry D, Jarvi S, Nutman TB.Angiostrongylus cantonensis (Ac), or the rat lungworm, is a major cause of eosinophilic meningitis. Humans are infected by ingesting the 3rd stage larvae from primary hosts, snails, and slugs, or paratenic hosts. The currently used molecular test is a qPCR assay targeting the ITS1 rDNA region (ITS1) of Ac. In silico design of a more sensitive qPCR assay was performed based on tandem repeats predicted to be the most abundant by the RepeatExplorer algorithm. Genomic DNA (gDNA) of Ac were used to determine the analytical sensitivity and specificity of the best primer/probe combination. This assay...
Alhassan A, Thekisoe OM, Yokoyama N, Inoue N, Motloang MY, Mbati PA, Yin H, Katayama Y, Anzai T, Sugimoto C, Igarashi I.Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid method whereby DNA is amplified with high specificity, efficiency, and rapidity under isothermal conditions using a set of four specifically designed primers and a DNA polymerase with strand displacement activity. In this study, we used LAMP primer sets designed from EMA-1 and Bc 48 genes for detection of Theileria equi and Babesia caballi infections, respectively. These primer sets specifically amplified DNA of the respective parasites. Both primer sets amplified T. equi and B. caballi up to 10(-6) dilution of 10-fold seria...
Wang L, Raidal SL, Pizzirani A, Wilcox GE.A nested multiplex PCR was developed as a rapid (<12h), sensitive test for the simultaneous identification of equine herpesviruses (EHV1, EHV4, EHV2 and EHV5) in clinical samples from horses. Peripheral blood and nasal swab (NS) samples from 205 weanling Thoroughbred foals on 6 different studs over 3 consecutive seasons and from 92 adult horses without clinical signs of respiratory disease were examined using direct multiplex PCR of clinical samples (direct PCR) and conventional cell culture with differentiation of EHV in cell cultures by multiplex PCR. Multiplex PCR proved a sensitive and ...
Birhanu H, Fikru R, Said M, Kidane W, Gebrehiwot T, Hagos A, Alemu T, Dawit T, Berkvens D, Goddeeris BM, Büscher P.African animal trypanosomosis, transmitted cyclically by tsetse flies or mechanically by other biting flies, causes serious inflictions to livestock health. This study investigates the extent of non-tsetse transmitted animal trypanosomosis (NTTAT) by Trypanosoma (T.) evansi and T. vivax in domestic animals in the tsetse-free regions of Northern Ethiopia, Afar and Tigray. Methods: A cross sectional study was conducted on 754 dromedary camels, 493 cattle, 264 goats, 181 sheep, 84 donkeys, 25 horses and 10 mules. The microhaematocrit centrifugation technique was used as parasitological test. Plas...
Kim CM, Blanco LB, Alhassan A, Iseki H, Yokoyama N, Xuan X, Igarashi I.We developed a TaqMan real-time polymerase chain reaction (PCR) assay for the quantitative detection of Theileria equi from the in vitro-cultured parasite and field blood samples collected from horses living in Ghana and Brazil. The detection limit for the assay was determined to be 1.5 parasites/microl per sample, and the quantitative capacity was demonstrated using the in vitro-cultured parasite. For field applications, the real-time PCR assay was compared to a previously established nested PCR assay used as the gold standard for the real-time PCR assay. Of 65 field blood samples, 46 samples...
Tomlinson JE, Kapoor A, Kumar A, Tennant BC, Laverack MA, Beard L, Delph K, Davis E, Schott Ii H, Lascola K, Holbrook TC, Johnson P, Taylor SD....Three flaviviruses (equine pegivirus [EPgV]; Theiler's disease-associated virus [TDAV]; non-primate hepacivirus [NPHV]) and equine parvovirus (EqPV-H) are present in equine blood products; the TDAV, NPHV, and EqPV-H have been suggested as potential causes of serum hepatitis. Objective: To determine the prevalence of these viruses in horses with equine serum hepatitis. Methods: Eighteen horses diagnosed with serum hepatitis, enrolled from US referral hospitals. Methods: In the prospective case study, liver, serum, or both samples were tested for EPgV, TDAV, NPHV, and EqPV-H by PCR. Results: Bot...
Qi M, Wang R, Wang H, Jian F, Li J, Zhao J, Dong H, Zhu H, Ning C, Zhang L.In present study, 262 fecal specimens were collected from 12 groups of grazing horses in the Xinjiang Uyghur Autonomous Region of China. The specimens were subjected to PCR and sequencing analyses of the ribosomal internal transcribed spacer (ITS). The overall prevalence of E. bieneusi in horses was 30.9% (81/262). No significant differences in prevalence were observed between horses of different ages or sexes. Nineteen genotypes were identified: 15 known genotypes (BEB6, CHG19, CM6, CM7, CM8, CS-1, CS-4, D, EpbA, EbpC, G, horse1, horse2, O, and Peru8) and four new genotypes (XJH1-XJH4). Six ...
Bermúdez CS, Zaldívar AY, Spolidorio MG, Moraes-Filho J, Miranda RJ, Caballero CM, Mendoza Y, Labruna MB.The present research evaluated the presence of Rickettsia spp. on ectoparasites of horses and dogs (using PCR techniques), and their sera (using immunofluorescence assay) in El Valle de Antón town in Panama. A total of 20 horses and 20 dogs were sampled, finding four species of ectoparasites on dogs (the ticks Rhipicephalus sanguineus, Amblyomma ovale, Amblyomma oblongoguttatum, and the flea Ctenocephalides felis), and two tick species on horses (Amblyomma cajennense and Dermacentor nitens). DNA of Rickettsia amblyommii was found in pools of A. cajennense, D. nitens, and R. sanguineus, while ...
Nielsen MK, Peterson DS, Monrad J, Thamsborg SM, Olsen SN, Kaplan RM.Strongylus vulgaris is an important strongyle nematode with high pathogenic potential infecting horses world-wide. Several decades of intensive anthelmintic use has virtually eliminated clinical disease caused by S. vulgaris, but has also caused high levels of anthelmintic resistance in equine small strongyle (cyathostomin) nematodes. Recommendations aimed at limiting the development of anthelmintic resistance by reducing treatment intensity raises a simultaneous demand for reliable and accurate diagnostic tools for detecting important parasitic pathogens. Presently, the only means available t...
Hulínská D, Langrová K, Pejcoch M, Pavlásek I.The aim of this study was to detect Anaplasma phagocytophilum in wild and domesticated animals and to identify the phylogenetic relationships of different strains of this bacterium. We adapted six published conventional methods targeting 16S fragments for real-time polymerase chain reaction. Initial screening of samples from 419 animals found 37 Anaplasma positives, later confirmed with several different primers and a TaqMan probe. We also performed DNA quantification and melting curve analysis. The nucleic acid of Anaplasma sp. was detected in a higher percentage of cases in members of the de...
Campbell AJ, Gasser RB, Chilton NB.In the current study, molecular techniques were evaluated for the species identification of individual strongyle eggs. Adult worms of Strongylus edentatus, S. equinus and S. vulgaris were collected at necropsy from horses from Australia and the U.S.A. Genomic DNA was isolated and a ribosomal transcribed spacer (ITS-2) amplified and sequenced using polymerase chain reaction (PCR) techniques. The length of the ITS-2 sequence of S. edentatus, S. equinus and S. vulgaris ranged between 217 and 235 nucleotides. Extensive sequence analysis demonstrated a low degree (0-0.9%) of intraspecific variation...
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...
Diallo IS, Hewitson G, Wright L, Rodwell BJ, Corney BG.Equid herpesvirus 1 (EHV1) is a major disease of equids worldwide causing considerable losses to the horse industry. A variety of techniques, including PCR have been used to diagnose EHV1. Some of these PCRs were used in combination with other techniques such as restriction enzyme analysis (REA) or hybridisation, making them cumbersome for routine diagnostic testing and increasing the chances of cross-contamination. Furthermore, they involve the use of suspected carcinogens such as ethidium bromide and ultraviolet light. In this paper, we describe a real-time PCR, which uses minor groove-bindi...
Bashiruddin JB, Cammà C, Rebêlo E.Babesia equi and Babesia caballi are tick-borne haemoparasites that may cause babesiosis of Equidae. In southern Europe B. equi is enzootic and infections may occur asymptomatically and more frequently than those due to B. caballi. Complement fixation test (CFT) is the official serological test for the diagnosis of equine babesiosis, but it has low sensitivity during early and latent stages of the disease. With the aim of developing more sensitive and rapid direct diagnostic alternatives, PCR systems that amplified DNA targets of 664 or 659 bp regions of the 16S rRNA genes were designed and de...
Fenger CK, Granstrom DE, Langemeier JL, Stamper S, Donahue JM, Patterson JS, Gajadhar AA, Marteniuk JV, Xiaomin Z, Dubey JP.Sarcocystis neurona is an apicomplexan that causes equine protozoal myeloencephalitis (EPM) in North and South America. Horses appear to be an aberrant host, because the merozoites continually divide in the central nervous system, without encysting. The natural host species has not previously been identified. The small subunit ribosomal RNA (SSURNA) gene of S. neurona was compared to those of Sarcocystis muris, Sarcocystis cruzi, Toxoplasma gondii, and Cryptosporidium parvum to identify a unique region suitable for a species-specific amplification primer. The S. neurona SSURNA primer was used ...
