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Topic:Infectious Disease

Infectious diseases in horses encompass a range of illnesses caused by bacteria, viruses, fungi, or parasites. These diseases can affect various systems within the equine body, leading to symptoms that range from mild discomfort to severe systemic illness. Common infectious diseases in horses include equine influenza, strangles, equine herpesvirus, and West Nile virus. These diseases can be transmitted through direct contact with infected animals, contaminated surfaces, or vectors such as insects. Understanding the mechanisms of transmission, pathogenesis, and immune response is essential for effective prevention and control. This page compiles peer-reviewed research studies and scholarly articles that explore the epidemiology, diagnosis, treatment, and management of infectious diseases in horses.
Glycoprotein H and α4β1 integrins determine the entry pathway of alphaherpesviruses.
Journal of virology    March 20, 2013   Volume 87, Issue 10 5937-5948 doi: 10.1128/JVI.03522-12
Azab W, Lehmann MJ, Osterrieder N.Herpesviruses enter cells either by direct fusion at the plasma membrane or from within endosomes, depending on the cell type and receptor(s). We investigated two closely related herpesviruses of horses, equine herpesvirus type 1 (EHV-1) and EHV-4, for which the cellular and viral determinants routing virus entry are unknown. We show that EHV-1 enters equine epithelial cells via direct fusion at the plasma membrane, while EHV-4 does so via an endocytic pathway, which is dependent on dynamin II, cholesterol, caveolin 1, and tyrosine kinase activity. Exchange of glycoprotein H (gH) between EHV-1...
Indirect fluorescent antibody test and surface antigen ELISAs for antemortem diagnosis of equine protozoal myeloencephalitis.
Journal of veterinary internal medicine    March 20, 2013   Volume 27, Issue 3 596-599 doi: 10.1111/jvim.12061
Johnson AL, Morrow JK, Sweeney RW.Recent research suggests that serum : CSF titer ratios could provide the most accurate antemortem diagnosis of equine protozoal myeloencephalitis (EPM) caused by Sarcocystis neurona. Objective: The purpose of this study was to assess the utility of two commercially available tests, the indirect fluorescent antibody test (IFAT) and the surface antigen 2, 4/3 ELISA (SAG2, 4/3 ELISA), using archived paired serum and CSF samples. Methods: Samples were obtained from 4 types of clinical patients. Confirmed positive cases (n = 9 horses; 11 sample sets) had neurologic deficits and postmortem lesions c...
Concurrent occurrence of human and equine West Nile virus infections in Central Anatolia, Turkey: the first evidence for circulation of lineage 1 viruses.
International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases    March 19, 2013   Volume 17, Issue 7 e546-e551 doi: 10.1016/j.ijid.2013.02.005
Ozkul A, Ergunay K, Koysuren A, Alkan F, Arsava EM, Tezcan S, Emekdas G, Hacioglu S, Turan M, Us D.West Nile fever is an important zoonotic infection caused by West Nile virus (WNV), a member of the Flaviviridae. Previous serological data from Turkey suggest widespread WNV circulation. This report includes cases of human and equine WNV infections occurring concurrently, and manifesting as central nervous system infections, in two neighboring provinces of Central Anatolia, Turkey. A partial phylogenetic analysis of the causative virus is given for the first time. Methods: The cases were reported in February (horses) and March (human). Symptoms of the disease were similar in the two species, ...
Diagnosis and isolation of Toxoplasma gondii in horses from Brazilian slaughterhouses. Evers F, Garcia JL, Navarro IT, Zulpo DL, Nino Bde S, Ewald MP, Pagliari S, Almeida JC, Freire RL.This study aimed to investigate anti-Toxoplasma gondii antibodies and to isolate the parasite from the brains of horses processed at slaughterhouses in Brazil. We collected brain and blood samples from 398 horses of various ages, from six Brazilian states. Serum samples were evaluated by indirect fluorescent antibody test (IFAT cut-off titre ≥ 1:64), and brains were submitted to mouse bioassay. Among the 398 horses, positivity for T. gondii was identified in 46 (11.6%) by IFAT and in 14 (3.5%) by mouse bioassay. In 12 of those 14 bioassays, mice were positive only by IFAT (cut-off titre ≥ ...
Monitoring of West Nile virus infections in Germany.
Zoonoses and public health    March 19, 2013   Volume 59 Suppl 2 95-101 doi: 10.1111/zph.12015
Ziegler U, Seidowski D, Angenvoort J, Eiden M, Müller K, Nowotny N, Groschup MH.West Nile virus (WNV) is a flavivirus that is maintained in an enzootic cycle between ornithophilic mosquitoes, mainly of the Culex genus, and certain wild bird species. Other bird species like ravens, jays and raptors are highly susceptible to the infection and may develop deadly encephalitis, while further species of birds are only going through subclinical infection. The objective of this study was to continue in years 2009-2011 the serological and molecular surveillance in wild birds in Germany (see Vector Borne Zoonotic Dis. 10, 639) and to expand these investigations for the first time a...
Identification of a previously undescribed divergent virus from the Flaviviridae family in an outbreak of equine serum hepatitis.
Proceedings of the National Academy of Sciences of the United States of America    March 18, 2013   Volume 110, Issue 15 E1407-E1415 doi: 10.1073/pnas.1219217110
Chandriani S, Skewes-Cox P, Zhong W, Ganem DE, Divers TJ, Van Blaricum AJ, Tennant BC, Kistler AL.Theiler's disease is an acute hepatitis in horses that is associated with the administration of equine blood products; its etiologic agent has remained unknown for nearly a century. Here, we used massively parallel sequencing to explore samples from a recent Theiler's disease outbreak. Metatranscriptomic analysis of the short sequence reads identified a 10.5-kb sequence from a previously undescribed virus of the Flaviviridae family, which we designate "Theiler's disease-associated virus" (TDAV). Phylogenetic analysis clusters TDAV with GB viruses of the recently proposed Pegivirus genus, altho...
Imaging diagnosis-aortic aneurysm and ureteral obstruction secondary to umbilical artery abscessation in a 5-week-old foal. Nogradi N, Magdesian KG, Whitcomb MB, Church M, Spriet M.A 5-week-old foal was evaluated for fever and hematuria of 3 days duration. Cystoscopy localized the blood to be originating from the left ureter. Abdominal ultrasonography revealed left hydronephrosis, hydroureter, and omphaloarteritis of the left umbilical artery with abscess formation that communicated with an arterial structure. Computed tomography (CT) revealed a large aortic aneurysm within the center of the abscess. An exploratory celiotomy was performed and the infection was nonresectable. The prognosis for life was grave; therefore the colt was euthanized. Necropsy findings confirmed ...
Prevalence and risk factors for Equine Infectious Anemia in Poconé municipality, northern Brazilian Pantanal.
Research in veterinary science    March 18, 2013   Volume 95, Issue 1 76-81 doi: 10.1016/j.rvsc.2013.02.011
Borges AM, Silva LG, Nogueira MF, Oliveira AC, Segri NJ, Ferreira F, Witter R, Aguiar DM.Serum samples collected from 547 equids in the Pantanal region of Brazil were evaluated for antibodies to Equine Infectious Anemia Virus (EIAV) by the agar gel immunodiffusion test. Risk factors associated with EIAV seropositivity were evaluated and spatial dependence investigated using a Spatial Lag Model. EIAV prevalence on farms in the Pantanal was 52.0% (13/25) with adjusted prevalence between equids of 31.5% (17.4-48.8% 95% CI). Intra-herd prevalence ranged from 5.0 to 77.0%. Statistical analysis demonstrated that farms and animals in regularly flooded areas had respectively 60 and 146 fo...
Untangling the complex inter-relationships between horse managers’ perceptions of effectiveness of biosecurity practices using Bayesian graphical modelling.
Preventive veterinary medicine    March 13, 2013   Volume 110, Issue 1 37-44 doi: 10.1016/j.prevetmed.2013.02.004
Schemann K, Lewis FI, Firestone SM, Ward MP, Toribio JA, Taylor MR, Dhand NK.On-farm biosecurity practices have been promoted in many animal industries to protect animal populations from infections. Current approaches based on regression modelling techniques for assessing biosecurity perceptions and practices are limited for analysis of the interrelationships between multivariate data. A suitable approach, which does not require background knowledge of relationships, is provided by Bayesian network modelling. Here we apply such an approach to explore the complex interrelationships between the variables representing horse managers' perceptions of effectiveness of on-far...
Knowledge base and effectiveness of online continuing education about foreign animal diseases for equine veterinarians.
Journal of veterinary medical education    March 12, 2013   Volume 40, Issue 1 69-75 doi: 10.3138/jvme.0612-056R
Wiedenheft AM, Traub-Dargatz JL, Gillette SC, O'Keefe GJ, Rao S, Salman MD.The purpose of this study was to assess the effectiveness of two different methods of online education using the knowledge base of African horse sickness (AHS) among US equine veterinarians as a model. An e-mail was sent to US veterinary members of the American Association of Equine Practitioners (AAEP), inviting them to participate in a complementary online educational opportunity. We determined participants' baseline knowledge of AHS by their responses in an AHS case scenario. Participants were then randomly assigned to either a Webinar module or a text-formatted module, followed by an educa...
Equine keratomycosis in Switzerland: a retrospective evaluation of 35 horses (January 2000-August 2011).
Equine veterinary journal    March 12, 2013   Volume 45, Issue 5 608-612 doi: 10.1111/evj.12042
Voelter-Ratson K, Pot SA, Florin M, Spiess BM.Keratomycosis is a severe disease in horses. Geographical differences in fungi causing keratomycosis and susceptibility of the organisms to antifungal drugs exist but few previous publications on this disease originate from Europe. Objective: To retrospectively compare the clinical data of 36 eyes with keratomycosis, diagnosed in 35 horses between January 2000 and August 2011 at the Vetsuisse Faculty of Switzerland. Case history, season, prior treatment, clinical appearance, surgical and medical treatment, treatment duration, and globe survival were evaluated. Methods: Retrospective case serie...
Equine herpesvirus type 1 infection induces procoagulant activity in equine monocytes.
Veterinary research    March 11, 2013   Volume 44, Issue 1 16 doi: 10.1186/1297-9716-44-16
Yeo WM, Osterrieder N, Stokol T.The alphaherpesvirus, equine herpesvirus type 1 (EHV-1), is a highly prevalent cause of equine infectious abortion and encephalomyelopathy. These syndromes have been attributed to ischemic necrosis from thrombosis in placental and neural vessels, although the mechanisms underlying thrombosis are unknown. After inhalation, EHV-1 establishes a peripheral blood mononuclear cell-associated viremia, with monocytes being a target of infection. Monocytes are also the main source of tissue factor (TF) in diseased states. Since TF is the primary activator of coagulation, increased monocyte TF expressio...
Molecular epidemiology of Theileria equi in horses and their association with possible tick vectors in the state of Rio de Janeiro, Brazil.
Parasitology research    March 9, 2013   Volume 112, Issue 5 2017-2025 doi: 10.1007/s00436-013-3360-0
Peckle M, Pires MS, Dos Santos TM, Roier EC, da Silva CB, Vilela JA, Santos HA, Massard CL.The aim of this study was to detect Theileria equi (Laveran 1901) DNA in horses and ticks using real-time PCR and to list the factors associated with infection in animals located in the Seropedica and Petropolis municipalities of the state of Rio de Janeiro. We tested blood samples from 314 horses and samples from 300 ticks, including 191 Amblyomma cajennense, 104 Dermacentor nitens, and 5 Ixodida larvae. Factors inherent to the horse, the ownership, and animal management were obtained from an epidemiological questionnaire and were evaluated in association with the presence of T. equi DNA in t...
The nasal vestibulum is the optimal sampling site for MRSA screening in hospitalised horses.
Veterinary journal (London, England : 1997)    March 7, 2013   Volume 197, Issue 2 415-419 doi: 10.1016/j.tvjl.2013.01.031
Van den Eede A, Hermans K, Van den Abeele A, Floré K, Dewulf J, Vanderhaeghen W, Némeghaire S, Butaye P, Gasthuys F, Haesebrouck F, Martens A.The increased incidence of methicillin-resistant Staphylococcus aureus (MRSA) infection in equine hospitals highlights the need for infection control protocols based on optimal patient screening. In horses, the deep ventral meatus of the nasal cavity is the principal site sampled to detect MRSA. However, in humans, the anterior nares are the preferred sampling site. The objective of this study was to determine the optimal sampling location in the nasal chambers for MRSA in horses by comparing the results obtained from three different locations (the vestibulum, diverticulum and ventral meatus) ...
Understanding the associations between on-farm biosecurity practice and equine influenza infection during the 2007 outbreak in Australia.
Preventive veterinary medicine    March 6, 2013   Volume 110, Issue 1 28-36 doi: 10.1016/j.prevetmed.2013.02.003
Firestone SM, Lewis FI, Schemann K, Ward MP, Toribio JA, Dhand NK.In a previous premises-level case-control study of the 2007 equine influenza outbreak in Australia, the protective effect of several variables representing on-farm biosecurity practices was identified. However, using logistic regression it was not possible to definitively identify individual effects and associations between each of the personal biosecurity measures implemented by horse premises owners and managers in the face of the outbreak. In this study we apply Bayesian network modelling to identify the complex web of associations between these variables, horse premises infection status an...
Venezuelan equine encephalitis replicon particles can induce rapid protection against foot-and-mouth disease virus.
Journal of virology    March 6, 2013   Volume 87, Issue 10 5447-5460 doi: 10.1128/JVI.03462-12
Diaz-San Segundo F, Dias CC, Moraes MP, Weiss M, Perez-Martin E, Owens G, Custer M, Kamrud K, de los Santos T, Grubman MJ.We have previously shown that delivery of the porcine type I interferon gene (poIFN-α/β) with a replication-defective human adenovirus vector (adenovirus 5 [Ad5]) can sterilely protect swine challenged with foot-and-mouth disease virus (FMDV) 1 day later. However, the need of relatively high doses of Ad5 limits the applicability of such a control strategy in the livestock industry. Venezuelan equine encephalitis virus (VEE) empty replicon particles (VRPs) can induce rapid protection of mice against either homologous or, in some cases, heterologous virus challenge. As an alternative approach ...
Detection of Theileria equi and Babesia caballi infections in Venezuelan horses using Competitive-Inhibition ELISA and PCR.
Veterinary parasitology    March 5, 2013   Volume 196, Issue 1-2 37-43 doi: 10.1016/j.vetpar.2013.02.004
Rosales R, Rangel-Rivas A, Escalona A, Jordan LS, Gonzatti MI, Aso PM, Perrone T, Silva-Iturriza A, Mijares A.The focus of this study was the detection of equine piroplasmosis in Distrito Capital, Miranda, Aragua, Guárico and Apure States from Venezuela, using two methods: Competitive-Inhibition ELISA and multiplex PCR and the analysis of the possible differences in occurrence in relation to the primary purpose of the horses, which is related to varied degrees of exposure to tick. Antibody levels to Babesia caballi and Theileria equi were assessed in 694 equine serum samples using Competitive-Inhibition ELISA, while PCR assays were performed in 136 horses, using two sets of oligonucleotides to establ...
Clinical observations and management of a severe equine herpesvirus type 1 outbreak with abortion and encephalomyelitis.
Acta veterinaria Scandinavica    March 5, 2013   Volume 55, Issue 1 19 doi: 10.1186/1751-0147-55-19
Walter J, Seeh C, Fey K, Bleul U, Osterrieder N.Latent equine herpesvirus type 1 (EHV-1) infection is common in horse populations worldwide and estimated to reach a prevalence nearing 90% in some areas. The virus causes acute outbreaks of disease that are characterized by abortion and sporadic cases of myeloencephalopathy (EHM), both severe threats to equine facilities. Different strains vary in their abortigenic and neuropathogenic potential and the simultaneous occurrence of EHM and abortion is rare. In this report, we present clinical observations collected during an EHV-1 outbreak caused by a so-called "neuropathogenic" EHV-1 G(2254)/D(...
Eliminating infectious diseases of livestock: a metapopulation model of infection control.
Theoretical population biology    March 4, 2013   Volume 85 63-72 doi: 10.1016/j.tpb.2013.02.002
Glass K, Barnes B.When novel disease outbreaks occur in livestock, policy makers must respond promptly to eliminate disease, and are typically called on to make control decisions before detailed analysis of disease parameters can be undertaken. We present a flexible metapopulation model of disease spread that incorporates variation in livestock density and includes occasional high-mixing locations or events, such as markets or race meetings. Using probability generating functions derived from this branching process model, we compare the likely success of reactive control strategies in eliminating disease spread...
Retrospective study on equine uterine fungal isolates and antifungal susceptibility patterns (1999-2011).
Equine veterinary journal. Supplement    March 2, 2013   Issue 43 84-87 doi: 10.1111/j.2042-3306.2012.00608.x
Beltaire KA, Cheong SH, Coutinho da Silva MA.Knowledge of commonly encountered fungi infecting the mare's reproductive tract and their respective drug susceptibilities should improve treatment efficacy in mares with fungal endometritis. This is particularly important when practitioners need to start empiric treatment before culture results are complete. Objective: To report the spectrum of fungal isolates from uterine samples from mares with reproductive problems and their respective antifungal susceptibilities. Methods: Equine uterine samples submitted to the Cornell University Animal Health Diagnostic Centre for fungal culture between ...
Surgical treatment for acute small intestinal obstruction caused by Parascaris equorum infection in 15 horses (2002-2011).
Equine veterinary journal. Supplement    March 2, 2013   Issue 43 111-114 doi: 10.1111/j.2042-3306.2012.00607.x
Tatz AJ, Segev G, Steinman A, Berlin D, Milgram J, Kelmer G.There is a consensus in the veterinary literature that Parascaris equorum (ascarid) impaction carries a poor prognosis. Hence surgery is often delayed and foals are often subjected to euthanasia after diagnosis of ascarid impaction is established. Our clinical impression was that ascarid impaction carries a better prognosis than previously reported. Objective: Our expectation was that manually evacuating the impaction into the caecum, thus refraining from opening the small intestine, would improve the prognosis for survival of horses with ascarid impaction. The aim of this study was to examine...
Effects of repeated transvaginal aspiration of immature follicles on mare health and ovarian status.
Equine veterinary journal. Supplement    March 2, 2013   Issue 43 78-83 doi: 10.1111/j.2042-3306.2012.00606.x
Velez IC, Arnold C, Jacobson CC, Norris JD, Choi YH, Edwards JF, Hayden SS, Hinrichs K.Transvaginal ultrasound-guided follicle aspiration (TVA) is performed clinically but there is little information available on complications associated with this procedure. Objective: It is possible that TVA is associated with damage to the ovary and may induce peritonitis or peritoneal adhesions. This study was conducted to determine the effect of repeated TVA on mare health and ovarian status. Methods: Thirty-two mares were used for oocyte recovery via repeated TVA over a 3 year period; different mares were used each year. In Year 1, ovarian status was monitored in 11 mares by transrectal pal...
B-mode and Doppler ultrasonography in pony mares with experimentally induced ascending placentitis.
Equine veterinary journal. Supplement    March 2, 2013   Issue 43 88-94 doi: 10.1111/j.2042-3306.2012.00658.x
Bailey CS, Heitzman JM, Buchanan CN, Bare CA, Sper RB, Borst LB, Macpherson M, Archibald K, Whitacre M.Early, accurate diagnosis of ascending placentitis in mares remains a key challenge for successful treatment of the disease. Doppler ultrasonography has shown promise as a tool to diagnose pregnancy abnormalities and is becoming more available to equine clinicians. However, to date, no studies have prospectively compared this technique to standard B-mode measurement of the combined thickness of the uterus and placenta (CTUP). Objective: The objective of the current study was to compare Doppler and B-mode ultrasonography for the detection of experimentally-induced ascending placentitis in mares...
Detection of anti-Toxoplasma gondii antibodies in carthorses in the metropolitan region of Curitiba, Paraná, Brazil. Finger MA, Villalobos EM, Lara Mdo C, Cunha EM, Barros Filho IR, Deconto I, Dornbusch PT, Ullmann LS, Biondo AW.Toxoplasma gondii, the agent for toxoplasmosis, has worldwide distribution. Horses normally play a secondary role in its life cycle, but movement around urban areas, feeding on grass and the increasing use of carthorses for gathering recyclable material in some urban areas of Brazil may increase their exposure to T. gondii infection. The aim of the present study was to investigate the frequency of anti-T. gondii antibodies in carthorses in the metropolitan region of Curitiba, PR. IgG antibodies against T. gondii were detected using the indirect fluorescence antibody test (IFAT) (titers ≥ 64)...
Combining two serological assays optimises sensitivity and specificity for the identification of Streptococcus equi subsp. equi exposure.
Veterinary journal (London, England : 1997)    March 1, 2013   Volume 197, Issue 2 188-191 doi: 10.1016/j.tvjl.2013.01.033
Robinson C, Steward KF, Potts N, Barker C, Hammond TA, Pierce K, Gunnarsson E, Svansson V, Slater J, Newton JR, Waller AS.The detection of anti-Streptococcus equi antibodies in the blood serum of horses can assist with the identification of apparently healthy persistently infected carriers and the prevention of strangles outbreaks. The aim of the current study was to use genome sequencing data to develop an indirect enzyme linked immunosorbent assay (iELISA) that targets two S. equi-specific protein fragments. The sensitivity and specificity of the antigen A and antigen C iELISAs were compared to an SeM-based iELISA marketed by IDvet - diagnostic Vétérinaire (IDvet). Individually, each assay compromised specifi...
The evolutionary consequences of alternative types of imperfect vaccines.
Journal of mathematical biology    February 28, 2013   Volume 68, Issue 4 969-987 doi: 10.1007/s00285-013-0654-x
Magori K, Park AW.The emergence and spread of mutant pathogens that evade the effects of prophylactic interventions, including vaccines, threatens our ability to control infectious diseases globally. Imperfect vaccines (e.g. those used against influenza), while not providing life-long immunity, confer protection by reducing a range of pathogen life-history characteristics; conversely, mutant pathogens can gain an advantage by restoring the same range of traits in vaccinated hosts. Using an SEIR model motivated by equine influenza, we investigate the evolutionary consequences of alternative types of imperfect va...
Progressive control of equine infectious anaemia through more accurate diagnosis.
The Veterinary record    February 26, 2013   Volume 172, Issue 8 208-209 doi: 10.1136/vr.f1131
Maanen Cv.No abstract available
Current status of equine piroplasmosis in the Sudan.
Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases    February 26, 2013   Volume 16 191-199 doi: 10.1016/j.meegid.2013.02.008
Salim B, Bakheit MA, Kamau J, Sugimoto C.This is a cross-sectional molecular epidemiological study on equine piroplasmosis (EP) affecting horses and donkeys in the Sudan. The study evaluated 499 samples from geographically distinct regions in eastern, central and western parts of the country. PCR amplification of the 18S rRNA gene of both Thelieria equi and Babesia caballi was carried out. Horses from all sampled areas were found positive to T. equi DNA but no B. caballi was detected. Absence of B. caballi infection was confirmed by another PCR targeting the B. caballi 48-kDa merozoite antigen. The overall prevalence was found to be ...
Sequence analysis of ORFs 5, 6 and 7 of equine arteritis virus during persistent infection of the stallion–a 7-year study.
Veterinary microbiology    February 24, 2013   Volume 164, Issue 3-4 378-382 doi: 10.1016/j.vetmic.2013.02.011
Rola J, Socha W, Zmudzinski JF.Nucleotide and amino acid sequences of ORFs 5, 6 and 7 of EAV during persistent infection in the stallion of the Malopolska breed were analysed in the study. A total of 11 blood and semen samples were collected between 2004 and 2011. The titre of specific EAV antibodies in this carrier stallion was maintained at a high level throughout the study and was equal approximately 1:128. The sequence analysis of ORF5 showed 16 variable sites including 12 with synonymous substitutions and 4 with non-synonymous substitutions. The degree of nucleotide sequence identity among the strains ranged from 98.92...
Antibodies to OspC, OspF and C6 antigens as indicators for infection with Borrelia burgdorferi in horses.
Equine veterinary journal    February 22, 2013   Volume 45, Issue 5 533-537 doi: 10.1111/evj.12033
Wagner B, Goodman LB, Rollins A, Freer HS.Lyme disease is caused by Borrelia burgdorferi, which is transmitted by infected ticks (Ixodes spp.). Reports on Lyme disease in horses have increased in recent years. Nevertheless, the diagnosis of Lyme disease in horses is still challenging owing to its vague clinical presentation and the limitations of diagnostic tests. Objective: This study used a new serological Lyme multiplex assay to examine antibody responses to 3 antigens of B. burgdorferi, outer surface protein (Osp) C, OspF and C6, and to verify their use as markers for early and late infection stages in horses. Methods: Multiplex...