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Topic:Piroplasmosis

Piroplasmosis is a tick-borne disease affecting horses, caused by the protozoan parasites Theileria equi and Babesia caballi. These parasites invade and multiply within the red blood cells of the host, leading to hemolytic anemia. Clinical signs of piroplasmosis in horses may include fever, lethargy, jaundice, and anemia, although some horses may remain asymptomatic carriers. Diagnosis is typically confirmed through blood tests, including serological assays and polymerase chain reaction (PCR) techniques. Piroplasmosis can impact equine health and performance, and it poses challenges for international horse movement due to quarantine regulations. This page compiles peer-reviewed research studies and scholarly articles that explore the epidemiology, pathogenesis, diagnostic methods, and management strategies for equine piroplasmosis.
Prevalence of the causative agents of equine piroplasmosis in the South West of The Netherlands and the identification of two autochthonous clinical Theileria equi infections.
Veterinary journal (London, England : 1997)    January 21, 2012   Volume 193, Issue 2 381-385 doi: 10.1016/j.tvjl.2011.12.014
Butler CM, Sloet van Oldruitenborgh-Oosterbaan MM, Stout TA, van der Kolk JH, Wollenberg Lv, Nielen M, Jongejan F, Werners AH, Houwers DJ.Equine piroplasmosis (EP) has not been considered indigenous in The Netherlands. However, following the detection of an apparently indigenous subclinical Babesia caballi infection in a horse on Schouwen-Duiveland (an island in the Zeeland Province), a survey was undertaken between May and September 2010 to assess the prevalence of the causative agents of EP in the South-West of The Netherlands. Blood samples from 300 randomly selected horses were tested for specific antibodies against Theileria equi and B. caballi using an indirect fluorescence antibody test (IFAT), and for parasite DNA using ...
Demographic and environmental risk factors for infection by Theileria equi in 590 horses in Israel.
Veterinary parasitology    January 17, 2012   Volume 187, Issue 3-4 558-562 doi: 10.1016/j.vetpar.2012.01.018
Steinman A, Zimmerman T, Klement E, Lensky IM, Berlin D, Gottlieb Y, Baneth G.The prevalence of Theileria equi infection as well as the environmental and demographic risk factors for infection was studied in 590 healthy horses from 46 farms in Israel. The prevalence of T. equi DNA was assessed using a polymerase chain reaction for a segment of the Theileria 18S rRNA gene. The overall prevalence was 26.4% (156/590). There was a significant geographical variation in the prevalence of T. equi infection, ranging from 9.3% (25/270) in the central lowlands to 81.7% (49/60) in the Golan Heights. The prevalence of T. equi infection was found to be significantly associated with ...
Quest for the piroplasms in camels: identification of Theileria equi and Babesia caballi in Jordanian dromedaries by PCR.
Veterinary parasitology    November 29, 2011   Volume 186, Issue 3-4 456-460 doi: 10.1016/j.vetpar.2011.11.070
Qablan MA, Sloboda M, Jirků M, Oborník M, Dwairi S, Amr ZS, Hořín P, Lukeš J, Modrý D.DNA of two species of piroplasmids was detected in dromedaries during a survey of blood protozoans in Jordan between 2007 and 2009. Ten clinically healthy camels (10%) originating from three Jordanian districts were found, using a PCR assay, to harbor Theileria or Babesia species in their blood and no mix infection was determined. Analysis of the partial 18S rRNA gene sequences of these parasites allowed their unambiguous identification as equine piroplasmids Babesia caballi (n=6) and Theileria equi (n=4). In case of latter species, a novel genotype was found in horses. This first molecular-ba...
Equine piroplasmosis associated with Amblyomma cajennense Ticks, Texas, USA.
Emerging infectious diseases    October 18, 2011   Volume 17, Issue 10 1903-1905 doi: 10.3201/eid1710.101182
Scoles GA, Hutcheson HJ, Schlater JL, Hennager SG, Pelzel AM, Knowles DP.We report an outbreak of equine piroplasmosis in southern Texas, USA, in 2009. Infection prevalence reached 100% in some areas (292 infected horses). Amblyomma cajennense was the predominant tick and experimentally transmitted Theileria equi to an uninfected horse. We suggest that transmission by this tick species played a role in this outbreak.
Factors associated to Theileria equi in equids of two microregions from Rio de Janeiro, Brazil. Dos Santos TM, Roier EC, Santos HA, Pires MS, Vilela JA, Moraes LM, Almeida FQ, Baldani CD, Machado RZ, Massard CL.Serum samples from 714 equids of Itaguaí and Serrana microregions, Rio de Janeiro, southeastern Brazil, were examined by indirect fluorescent antibody test (titer 1:80) for Theileria equi. The prevalence in the microregions and factors associated with seropositivity were evaluated and the prevalence ratio (PR) calculated. The overall prevalence of T. equi infection was 81.09% (n = 579), with higher prevalence (p < 0.05) in the Itaguaí (85.43%) when compared to Serrana microregion (76.92%). The geographic area, altitude, farming condition and area of origin of equids were associated (p <...
Equine babesiosis: seroprevalence, risk factors and comparison of different diagnostic methods in Jordan.
Transboundary and emerging diseases    July 19, 2011   Volume 59, Issue 1 72-78 doi: 10.1111/j.1865-1682.2011.01244.x
Abutarbush SM, Alqawasmeh DM, Mukbel RM, Al-Majali AM.The purposes of this study were to estimate the seroprevalence and distribution of horse piroplasmosis, to evaluate risk factors associated with the occurrence of the disease and to compare the different diagnostic methods used for this disease. A total of 253 clinically normal horses were sampled, and a collection form was completed for each horse from five of six different climatic zones of Jordan. The sixth zone was not sampled because it did not include horse population. Competitive enzyme-linked immunosorbent assay (cELISA) revealed 37 horses (14.6%) positive for Theileria equi, and none ...
Identification of the piroplasms isolated from horses with clinical piroplasmosis in Poland.
Wiadomosci parazytologiczne    June 4, 2011   Volume 57, Issue 1 21-26 
Adaszek Ł, Górna M, Krzysiak M, Adaszek M, Garbal M, Winiarczyk S.The study was aimed at determining the cause of the diseases in three horses exhibiting symptoms of fever, ataxia, mucus membrane paleness, haematuria and thrombocytopenia. The PCR technique revealed the presence in the blood of 18S RNA Babesia/Theileria spp. genetic material. DNA amplification using primers RLB F2 and RLB R2 produced 430 bp size products. The sequences of these PCR products demonstrated a 95.6-97.5% similarity with the sequence of the fragment of 18S RNA Babesia equi, gene number DQ287951 in the GenBank. The treatment utilizing the subcutaneous application of the imidocarb re...
A survey for piroplasmids in horses and Bactrian camels in North-Eastern Mongolia.
Veterinary parasitology    March 12, 2011   Volume 179, Issue 1-3 246-249 doi: 10.1016/j.vetpar.2011.01.064
Sloboda M, Jirků M, Lukešová D, Qablan M, Batsukh Z, Fiala I, Hořín P, Modrý D, Lukeš J.Equine piroplasmosis caused by Babesia caballi and Theileria equi is widespread in Asia. The presence of these haemozoans in Mongolia was previously confirmed in domestic as well as in reintroduced Przewalski horses in which they cause significant pathology. The data on occurrence of piroplasms from Bactrian camels in Asia is lacking. A total of 192 horses, 70 Bactrian camels, and additional 16 shepherd dogs from the Hentiy province were included in our study. No clinical signs typical for piroplasmid infection were observed during the field survey. Microscopic examination revealed the presenc...
Seroprevalence of equine piroplasms in the Republic of Korea.
Veterinary parasitology    March 1, 2011   Volume 179, Issue 1-3 224-226 doi: 10.1016/j.vetpar.2011.02.020
Seo MG, Yun SH, Choi SK, Cho GJ, Park YS, Kwon OD, Cho KH, Kim TH, Jeong KS, Park SJ, Kwon YS, Kwak D.Equine piroplasms include two tick-borne protozoan parasites, Babesia caballi and Theileria equi. Although no clinical equine piroplasmosis has been reported in the Republic of Korea, the possible existence of the disease has been proposed due to a nationwide distribution of the vector ticks. To determine if the antibodies against B. caballi and T. equi were present, 184 sera of horses (Equus caballus) raised in the Republic of Korea from 2007 to 2010 were assessed using cELISA kits. Two (1.1%) out of 184 sera were positive for T. equi, but none were seropositive for B. caballi. Both samples t...
Prevalence of Theileria equi and Babesia caballi infection in horses from northern Italy.
Vector borne and zoonotic diseases (Larchmont, N.Y.)    January 23, 2011   Volume 11, Issue 7 955-956 doi: 10.1089/vbz.2010.0193
Grandi G, Molinari G, Tittarelli M, Sassera D, Kramer LH.Babesia caballi and Theileria equi are the causative agents of equine piroplasmosis. In this epidemiological study, 294 horses reared in a rural area of northern Italy were studied. During January 2008-January 2009, blood samples were taken for serology (indirect fluorescent antibody test) and for polymerase chain reaction (PCR). Data on the geographical area, sex, and age were collected for statistical analysis of risk factors associated with infection. A seroprevalence of 8.5% was found: 8.2% of the animals were positive for anti-T. equi antibodies and 0.3% for anti-B. caballi antibodies. No...
A case of transplacental transmission of Theileria equi in a foal in Trinidad.
Veterinary parasitology    October 20, 2010   Volume 175, Issue 3-4 363-366 doi: 10.1016/j.vetpar.2010.10.019
Georges KC, Ezeokoli CD, Sparagano O, Pargass I, Campbell M, D'Abadie R, Yabsley MJ.Equine piroplasmosis due to Theileria equi and Babesia caballi is endemic in Trinidad. A case of equine piroplasmosis due to T. equi was diagnosed in a thoroughbred foal at 10h post-partum. A high parasitaemia (63%) of piroplasms was observed in a Wright-Giemsa(®) stained thin blood smear from the foal. In addition, the 18S rRNA gene for Babesia/Theileria was amplified from DNA extracted from the blood of the foal and the mare. Amplified products were subjected to a reverse line blot hybridization assay (RLB), which confirmed the presence of T. equi DNA in the foal. The mare was negative by R...
Genetic diversity of piroplasms in plains zebra (Equus quagga burchellii) and Cape mountain zebra (Equus zebra zebra) in South Africa.
Veterinary parasitology    August 21, 2010   Volume 174, Issue 1-2 145-149 doi: 10.1016/j.vetpar.2010.08.014
Bhoora R, Buss P, Guthrie AJ, Penzhorn BL, Collins NE.Seventy EDTA blood samples collected from plains zebra (Equus quagga burchellii) and Cape mountain zebra (Equus zebra zebra) were screened for the presence of piroplasm parasite DNA using quantitative T. equi-specific and B. caballi-specific TaqMan real-time PCR (qPCR) tests. T. equi parasite DNA was detected in 60 samples, 19 of which were also positive for B. caballi. Approximately 1480bp of the piroplasm 18S rRNA gene was amplified and sequenced from 17 samples, while the V4 hypervariable region of the 18S rRNA gene was amplified, cloned and sequenced from 31 samples. BLASTN analysis reveal...
Geographic distribution modeling and spatial cluster analysis for equine piroplasms in Greece.
Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases    July 1, 2010   Volume 10, Issue 7 1013-1018 doi: 10.1016/j.meegid.2010.06.014
Kouam MK, Masuoka PM, Kantzoura V, Theodoropoulos G.Maximum entropy ecological niche modeling and spatial scan statistic were utilized to predict the geographic range and to investigate clusters of infections for equine piroplasms in Greece, using the Maxent and SaTScan programs, respectively. The eastern half of the country represented the culminating area with high probabilities (p>0.75) of presence of equine piroplasms and encompassed most regions with high concentration of equid host populations. The most important environmental factor that contributed to the ecological niche modeling was land cover followed by temperature. Significant c...
Genetic diversity of equine piroplasms in Greece with a note on speciation within Theileria genotypes (T. equi and T. equi-like).
Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases    June 19, 2010   Volume 10, Issue 7 963-968 doi: 10.1016/j.meegid.2010.06.008
Kouam MK, Kantzoura V, Masuoka PM, Gajadhar AA, Theodoropoulos G.Equine piroplasms in Greece were studied using the reverse line blot hybridization (RLB) assay. Three genotypes consisting of two Theileria (T. equi and T. equi-like) and one Babesia (B. caballi-like) were identified. Of 787 samples tested, 371 (47.14%) hybridised to catchall probe (probe specifically designed to capture any piroplasm species present in a sample), 346 (43.96%) to T. equi probe, 364 (46.25%) to T. equi-like probe, 0 (0%) to B. caballi probe and 3 (0.38%) to B. caballi-like probe. Seven samples gave faint signals with the catchall probe only, indicating the presence of known or ...
Sequence heterogeneity in the equi merozoite antigen gene (ema-1) of Theileria equi and development of an ema-1-specific TaqMan MGB assay for the detection of T. equi.
Veterinary parasitology    April 28, 2010   Volume 172, Issue 1-2 33-45 doi: 10.1016/j.vetpar.2010.04.025
Bhoora R, Quan M, Matjila PT, Zweygarth E, Guthrie AJ, Collins NE.Although a quantitative real-time PCR assay (qPCR) assay for the detection of Theileria equi has been developed and evaluated, it is possible that additional, as yet undetected 18S rRNA gene sequence variants may exist. A qPCR assay targeting a different gene, used in conjunction with the T. equi 18S rRNA qPCR assay, could assist in the detection of all T. equi genotypes in field samples. A T. equi ema-1-specific qPCR (Ueti et al., 2003) was tested on 107 South African field samples, 90 of which tested positive for T. equi antibody using the immuno-fluorescent antibody test (IFAT). The qPCR as...
Seroprevalence of Babesia caballi and Theileria equi in the Swiss horse population.
Parasitology international    February 21, 2010   Volume 59, Issue 3 313-317 doi: 10.1016/j.parint.2010.02.005
Sigg L, Gerber V, Gottstein B, Doherr MG, Frey CF.In Switzerland, the prevalence and incidence of equine piroplasma parasite (EPP) infections are unknown. In order to obtain a first insight into the prevalence, a representative sample of 689 sera of horses from Switzerland was serologically tested for the presence of antibodies directed against T. equi and B. caballi using the Indirect Fluorescence Antibody Test (IFAT). A total of 50 (7.3%) horses were seropositive for EPP: overall, the seroprevalence of T. equi was significantly higher than that of B. caballi (p=0.002). The seropositivities in indigenous horses (animals bred and raised in Sw...
Molecular detection of Theileria equi and Babesia caballi in the bone marrow of asymptomatic horses.
Veterinary parasitology    February 4, 2010   Volume 170, Issue 1-2 182-184 doi: 10.1016/j.vetpar.2010.01.043
Pitel PH, Pronost S, Scrive T, Léon A, Richard E, Fortier G.Equine piroplasmosis is a tick-borne disease, the aetiological agents of which are either Theileria equi or Babesia caballi parasites. Piroplasmosis is commonly encountered in acute or sub-acute clinical forms although clinically recovered horses may remain asymptomatic but infected for several years. The clinical detection of such apparently healthy carrier horses (that serve as a host for subsequent infecting ticks), remains a worldwide challenge for controlling the spread of the disease. The aim of the present paper is to report on the detection of both T. equi and B. caballi by PCR in the ...
Seroprevalence of equine piroplasms and host-related factors associated with infection in Greece.
Veterinary parasitology    January 20, 2010   Volume 169, Issue 3-4 273-278 doi: 10.1016/j.vetpar.2010.01.011
Kouam MK, Kantzoura V, Gajadhar AA, Theis JH, Papadopoulos E, Theodoropoulos G.Serum samples were collected from a total of 544 equids that included 524 horses, 13 mules, and 7 ponies from various regions of mainland Greece and were examined by competitive-inhibition ELISA (cELISA) to evaluate the level of exposure of Greek equids to Theileria (Babesia) equi and/or Babesia caballi, the causative agents of piroplasmosis. Association between seropositivity and host-related factors of species, gender, age, origin, activity and location were investigated. The overall seroprevalence was 11.6% (9.1-14.6%) with 95% confidence limit. The seroprevalence for T. equi and B. caballi...
Prevalence of equine Piroplasmosis and its association with tick infestation in the State of São Paulo, Brazil. Kerber CE, Labruna MB, Ferreira F, De Waal DT, Knowles DP, Gennari SM.Serum samples were collected from 582 horses from 40 stud farms in the State of São Paulo and tick (Acari: Ixodidae) infestations were evaluated on them. Serum samples were subjected to the complement fixation test (CFT) and a competitive inhibition ELISA (cELISA) for Babesia caballi and Theileria equi. Logistic regression analyses were performed to construct multivariate models that could explain the dependent variable (horses positive for B. caballi or T. equi) as a function of the independent variables (presence or abundance of each one of the tick species found on the farms). A higher ove...
Nucleotide sequence heterogeneity in the small subunit ribosomal RNA gene within Theileria equi from horses in Sudan.
Parasitology research    December 2, 2009   Volume 106, Issue 2 493-498 doi: 10.1007/s00436-009-1691-7
Salim B, Bakheit MA, Kamau J, Nakamura I, Sugimoto C.This is a molecular epidemiological investigation on Theileria equi, a causative agent of equine piroplasmosis. Blood samples were collected from 127 horses from different geographical locations in Sudan. The small subunit of rRNA gene (18S; ~1,600 bp) was amplified from 20 positive field samples and subsequently subjected to direct sequencing and analysis to reveal possible strain differences and the presence of a novel species or genotypes. Sequences were compared with published sequences mainly from South African and Spanish isolates. Eleven distinct T. equi sequences within 18S rRNA gene w...
A comparative study of serological tests and PCR for the diagnosis of equine piroplasmosis.
Parasitology research    November 6, 2009   Volume 106, Issue 3 709-713 doi: 10.1007/s00436-009-1669-5
Jaffer O, Abdishakur F, Hakimuddin F, Riya A, Wernery U, Schuster RK.A total of 105 serum samples from endurance horses from different stables in Dubai were examined for the presence of antibodies against Theileria equi and Babesia caballi using immunofluorescence antibody test (IFAT) and competitive enzyme-linked immunosorbent assay (cELISA). A TaqMan real-time polymerase chain reaction (PCR) was used to detect DNA of piroplasms in specimens of clotted blood or EDTA blood samples of the same animals. Out of the 105 serum samples, the IFAT detected antibodies against T. equi in 35 (33.3%) cases while the cELISA gave 34 (32.4%) positive results. Eleven (10.5%) o...
Immunogenicity and antigenicity of the recombinant EMA-1 protein of Theileria equi expressed in the yeast Pichia pastoris. Nizoli LQ, Conceição FR, Silva SS, Dummer LA, Santos AG, Leite FP.The equine piroplasmosis caused by Theileria equi is one of the most important parasitic diseases of the equine, causing damage to animal health and economic losses. In T. equi, 2 merozoite surface proteins, equi merozoite antigen EMA-1 and EMA-2, have been identified as the most immunodominant antigens. This suggests that these antigens might be used as immunobiological tools. The EMA-1 of Theileria equi was cloned and expressed in the yeast Pichia pastoris. The transformed yeast was grown at high cell density, expressing up to 389 mg x L(-1) of recombinant protein. The protein was concentrat...
Prevalence and diagnosis of Babesia and Theileria infections in horses in Italy: a preliminary study.
Veterinary journal (London, England : 1997)    April 24, 2009   Volume 184, Issue 3 346-350 doi: 10.1016/j.tvjl.2009.03.021
Moretti A, Mangili V, Salvatori R, Maresca C, Scoccia E, Torina A, Moretta I, Gabrielli S, Tampieri MP, Pietrobelli M.Babesia caballi and Theileria equi are the causative agents of equine piroplasmosis. In this preliminary epidemiological study, 412 horses reared in central and northern Italy were sampled and three diagnostic methods compared, namely, the microscopy, the indirect fluorescent antibody test (IFAT) and a PCR. Possible risk factors (such as area, season, breed, activity, sex, age, and grazing) associated with serological positivity were evaluated. A seroprevalence of 68.4% was found: 12.4% of the animals had anti-T. equi antibodies, 17.9% anti-B. caballi antibodies and 38.1% had antibodies agains...
A perspective on Theileria equi infections in donkeys.
The Japanese journal of veterinary research    April 11, 2009   Volume 56, Issue 4 171-180 
Kumar S, Kumar R, Sugimoto C.The donkey population has remained unchanged in the last two decades despite a decrease in the overall population of equids, emphasizing the usefulness of the donkey as a draught and pack animal. Piroplasmosis in donkeys, caused by Theileria equi and Babesia caballi, has been recognized as a serious problem of major economic importance as the affected animals manifest decreased working capacity, loss of appetite, etc. In tropical countries, T. equi infections are more wide-spread and pathogenic than those caused by B. caballi. Donkeys usually remain asymptomatic carriers with positive antibody...
Isolated fever in horses: a new case of equine anaplasmosis in France. Boni M, Rolain JM, Portelli C, Marié JM, Davoust B, Brouqui P.No abstract available
A molecular survey of Piroplasmida and Hepatozoon isolated from domestic and wild animals in Burgos (northern Spain).
Veterinary parasitology    February 26, 2009   Volume 162, Issue 1-2 147-150 doi: 10.1016/j.vetpar.2009.02.021
Gimenez C, Casado N, Criado-Fornelio A, de Miguel FA, Dominguez-Peñafiel G.This study reports a molecular survey of Hepatozoon species and of the order Piroplasmida in the Province of Burgos, northern Spain. The diagnostic techniques employed included PCR and the sequencing of the 18S rRNA gene. Eighty-nine blood samples from domestic animals plus 138 blood/coagulated blood samples from wild mammals were examined. Theilerid protozoa were found at relatively high frequencies in bovines (14.6%) and horses (36%). Theileria buffeli, T. sergenti and T. annulata were diagnosed in cows. T. equi was common in horses and T. annae was found in a donkey for the first time. A ne...
Development of an indirect ELISA for the diagnosis of equine piroplasmosis.
Annals of the New York Academy of Sciences    January 6, 2009   Volume 1149 235-238 doi: 10.1196/annals.1428.029
Asenzo G, Wilkowsky S, Barrandeguy M, Mesplet M, Benitez D, Florin-Christensen M.An indirect ELISA (iELISA) for the detection of specific anti-Theileria equi antibodies in horse serum was developed. Its performance showed good concordance (K= 0.79) when compared with a competitive ELISA recommended by the World Organisation for Animal Health. Horse serum samples from two provinces located in the north and east of Argentina (Formosa and Entre Rios, respectively) were analyzed by this iELISA. A high percentage of positive horses were found in Formosa, consistent with the climatic conditions of the region that are apt for the development of tick vectors. Surprisingly, seropos...
Exotic animal diseases bulletin: equine piroplasmosis.
Australian veterinary journal    December 9, 2008   Volume 86, Issue 11 N20-N21 
No abstract available
Transmission of Babesia caballi by Dermacentor nitens (Acari: Ixodidae) is restricted to one generation in the absence of alimentary reinfection on a susceptible equine host.
Journal of medical entomology    December 9, 2008   Volume 45, Issue 6 1152-1155 doi: 10.1603/0022-2585(2008)45[1152:tobcbd]2.0.co;2
Schwint ON, Knowles DP, Ueti MW, Kappmeyer LS, Scoles GA.The tropical horse tick, Dermacentor nitens, is a natural vector of Babesia caballi in the Americas. B. caballi, one of the etiologic agents of equine piroplasmosis, occurs widely throughout the world, but the United States and a few other countries are considered to be free of infection. B. caballi is transovarially transmitted by the one-host tick D. nitens; we tested the hypothesis that B. caballi can persist in multiple generations of D. nitens in the absence of opportunity to reacquire infection from a susceptible equine host. Partially engorged female D. nitens were collected from a B. c...
Sequence heterogeneity in the 18S rRNA gene within Theileria equi and Babesia caballi from horses in South Africa.
Veterinary parasitology    October 11, 2008   Volume 159, Issue 2 112-120 doi: 10.1016/j.vetpar.2008.10.004
Bhoora R, Franssen L, Oosthuizen MC, Guthrie AJ, Zweygarth E, Penzhorn BL, Jongejan F, Collins NE.A molecular epidemiological survey of the protozoal parasites that cause equine piroplasmosis was conducted using samples collected from horses and zebra from different geographical locations in South Africa. A total of 488 samples were tested for the presence of Theileria equi and/or Babesia caballi using the reverse line blot hybridization assay. Ten percent of the samples hybridized to the Theileria/Babesia genus-specific probe and not to the B. caballi or T. equi species-specific probes, suggesting the presence of a novel species or genotype. The small subunit of rRNA gene (18S; approximat...
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