Serology in horse research involves the study and analysis of blood serum to detect the presence of antibodies or antigens associated with infectious diseases and other health conditions. It is a diagnostic tool used to identify immune responses to pathogens, vaccination status, and exposure to specific diseases. Serological tests in equine research can include enzyme-linked immunosorbent assays (ELISA), complement fixation tests, and virus neutralization tests. These tests help in understanding the epidemiology of diseases, monitoring herd health, and informing vaccination strategies. This page compiles peer-reviewed research studies and scholarly articles that explore the methodologies, applications, and implications of serological testing in equine medicine.
Lee CL, Zandrew F, Davidsohn I.One hundred infectious mononucleosis and the same number of non-infectious mononucleosis sera were studied to evaluate the sensitivity and specificity of horse erythrocytes in the diagnosis of infectious mononucleosis. Titres of horse agglutinins in infectious mononucleosis sera ranged from 28 to 7,168 with a geometric mean of 550, whereas the corresponding sheep agglutinin titres ranged from less than 7 to 3,584, with a geometric mean of 126. Horse agglutinin titres of non-infectious mononucleosis sera ranged from less than 7 to 896, with a geometric mean of 59. Infectious mononucleosis sera ...
Morgante O, Vance HN, Shemanchuk JA, Windsor R.The epizootic of equine encephalomyelitis in 1965 in Alberta was proved to be due to Western Encephalomyelitis virus infection by serological findings and virus isolations.Sixty-three horses of 88 tested, showed a diagnostic rise of CF antibodies to Western Encephalomyelitis virus. Western Encephalomyelitis virus was isolated from 5 brains of horses. Homologous antibodies were shown in 3 of these animals, the only ones from which blood specimens were received. For the first time virological evidence is given that Western Encephalomyelitis virus infection in horses is found in more areas of Alb...
Busquets N, Gardela J, José-Cunilleras E, Solé A, Salvador MJ, Obón E, Molina-López R, Aranda C, Montalvo T, Corbella I, Bou-Monclús MA....West Nile virus (WNV), mainly transmitted by mosquitoes, poses significant health risks to humans and horses, particularly in endemic regions. The first detection of WNV lineage 2 in Spain was in 2017 in Catalonia (northeastern Spain). In 2023, WNV was confirmed in a young yellow-legged gull and a probable human case was notified within the urban settings. We aimed to define the zone of WNV circulation in the Barcelona Metropolitan Area where these infections occurred and the effectiveness of the One Health approach for early WNV detection. The Catalan WNV surveillance and control programs in...
Ganbaatar O, Batbaatar V, Kimura T, Ganzorig S, Tseren-Ochir EO, Suzuki Y, Takai S.ELISA antibody titers against Rhodococcus equi strain ATCC 6939 were measured in serum samples collected from 644 horses aged 1-21 years at 13 provinces and the capital of Mongolia, of which 631 (98%) samples showed values of approximately zero. The mean antibody titers in the 14 areas ranged from -0.08 to -0.01. Nine samples (1.4%) exhibited titers of 0.1-0.2, and surprisingly, four horses (0.6%) each aged 2, 5, 8, and 9 years demonstrated positive values in the range of 0.3-0.8. Considering the age resistance characteristic of this disease, the positive horses were hypothesized to exhibit no...
Motta D, Aymée L, Roussouliéres I, Di Azevedo MIN, Jacob JCF, Lilenbaum W.Leptospirosis is a zoonotic disease caused by spirochetes of the genus Leptospira. Equine genital leptospirosis (EGL) has been described as a chronic and silent syndrome, presenting reproductive alterations such as abortion, stillbirth, placentitis, embryonic loss, repeat breeding syndrome, and subfertility. This study aimed to investigate the genital Leptospira infection in naturally infected mares with poor reproductive performance, as well as to genetically characterize the agents. A total of 41 mares with a history of poor reproductive performance were selected. Sera were collected for ser...
Cardoso TL, Wozeak DR, Pereira IL, da Silva Ribeiro LD, Rodrigues RO, Hartwig DD.Leptospirosis is a globally distributed zoonosis that affects both humans and animals, with being the main causative agent. In horses, the disease is associated with considerable economic losses. The Microscopic Agglutination Test (MAT) is the reference test for diagnosis but has limitations, emphasizing the need for effective diagnostic alternatives. In this study, was evaluated the use of a recombinant chimera, composed of ErpY-like and LemA proteins, as an antigen for ELISA-based detection of equine leptospirosis. The chimera was successfully expressed, purified and tested on 915 horse ser...
Acevedo-Jiménez GE, Morales-González C, Akbarin MM, Rodríguez-Murillo C, González-Fernández VD, Avila-De la Vega LM, García-Flores MM....Equine infectious anemia virus (EIAV) is a globally significant equine pathogen, and its control relies on effective serological surveillance. The agar gel immunodiffusion (AGID) test, the WOAH-prescribed method, has limitations in sensitivity. This study developed and validated an indirect ELISA using a synthetic peptide derived from a conserved region of the EIAV gp90 surface glycoprotein. The assay was evaluated using 773 horse sera from the Metropolitan Zone of the Valley of Mexico and compared to the AGID test. The gp90-EIAV ELISA demonstrated a sensitivity of 85.3 % and a specificity o...
Badr C, Arbi M, Souiai O, Larbi I, Nsiri J, Elbehi I, Bouslama Z, Bennour MA, Essaied MS, Ines Khosrof , Chabchoub A, Ben Aoun B, Ghram A, Lachheb J.Equine influenza (EI) is a highly contagious viral respiratory disease affecting equids, with the potential of causing widespread outbreaks across Europe, the Americas, Asia, and other regions of the world. In Tunisia, in the spring of 2021, an EI virus outbreak occurred in a farm housing purebred Arabian horses that exhibited respiratory signs. This outbreak led to a national lockdown on horse movements and the cancellation of equestrian events. A total of 432 serum samples, collected from 2018 to 2021, were tested using ELISA competition assay. In addition, 100 Nasal swabs were collected dur...
Valderrama-Martinez C, Packham A, Smith W, Mendoza-Flores JE, Zheng S, Chigerwe M, Plancarte M, Aleman M.Long-term freezing storage can alter the stability of proteins, thereby compromising accurate determination of indirect fluorescent antibody test (IFAT) titers that support the diagnosis of equine protozoal myeloencephalitis. Objective: Assess the effect of long-term storage at -80°C on IFAT against S. neurona and N. hughesi in equine serum and cerebrospinal fluid (CSF). Methods: Paired serum and CSF (n = 46), and serum only (n = 25) samples. Methods: Prospective study of samples stored 6-12, 13-18, and 19-24 months. Comparing antibody titers across time points, McNemar and Wilcoxon...
Chinyere CN, Ajaebili AC, Peter-Ajuzie IK, Galadima HB, Daodu OB, Fatola OI, Okolo CC, Alaba BA, Akinniyi OO, Omoniwa DO, Edeh ER, Olorunfemi AB....African horse sickness (AHS) is a severe, infectious arthropod-borne disease of equids caused by the AHS virus (AHSV). It is endemic in Sub-Saharan Africa, and several sporadic outbreaks of the disease have been reported in Nigeria in the past 5 decades. Following a recent outbreak of the disease in Lagos State, this study was conducted to investigate the prevalence of (AHSV) antibodies in apparently healthy horses and some selected wildlife sampled in four geographical regions of Nigeria. Using a competitive enzyme-linked immunosorbent assay, 575 serum samples collected from horses in five ...
Nemoto M, Kawanishi N, Kamei R, Furusho K, Kawauchi K, Yabuuchi Y, Oue Y, Uchida Y, Nishiura H, Reedy SE, Chambers TM, Li F, Bannai H, Yamanaka T....In April and May 2025, outbreaks of equine influenza occurred for the first time in 17 years in Japan. Equine influenza virus (EIV) of the H3N8 subtype was mainly detected in heavy draft horse populations in Kumamoto Prefecture and the Tokachi area of Hokkaido. In total, 10 EIVs were isolated from infected horses and then were used for genetic and serological analyses. Phylogenetic analysis of all eight genes revealed that all Japanese isolates were clustered with the Florida sublineage clade 1 (Fc1) viruses and were closely related to North American Fc1 viruses detected in 2024-2025. The resu...
Şahinkesen İ, Bilge-Dağalp S.Equine herpesvirus 1 (EHV-1) and Equine herpesvirus 4 (EHV-4) are major viral pathogens of horses that contribute to respiratory disease, abortion, and neurological disorders, leading to economic losses in the equine industry. Accurate serological diagnosis is critical for disease surveillance and control. This study aimed to develop and validate an in-house enzyme-linked immunosorbent assay (ELISA) for detecting antibodies against EHV-1 and EHV-4 in horse sera. Serum samples previously confirmed by virus neutralization test (VNT) and a commercial ELISA were used to optimize the assay. Cut-off...
Carey KJ, Smith I, Hartley CA, Bailey KE, McIntosh K, Broder CC, Gilkerson JR.Hendra virus (HeV) is a high consequence pathogen that causes severe respiratory and/or neurological disease in both horses and humans with high mortality. The equine vaccine, Equivac® HeV, is a recombinant soluble version of the G glycoprotein of HeV (HeV-sG) that elicits a potent HeV-neutralising antibody response in adult horses and also provides indirect protection for humans. Maternally derived HeV-G specific antibodies could interfere with subsequent foal vaccination, although other factors such as foal age and the suitability of vaccination protocols that have been optimised for adult ...
McLinden LA, Kemp-Symonds JG, Daly JM, Blanchard AM, Waller AS, Freeman SL.Infection with Streptococcus equi subspecies equi (S. equi) is characterised by acute disease, with about 10% of infected animals remaining persistently infected. Clinically, infection with S. equi cannot readily be distinguished from infection caused by other respiratory pathogens, including Streptococcus equi subspecies zooepidemicus (S. zooepidemicus), equine influenza virus, and equine herpes virus. Screening protocols, with appropriate quarantining facilities, are important to detect horses infected with S. equi and avoid strangles outbreaks. Virulent strains of S. zooepidemicus can also ...
Kouadria W, Poder SL, van Maanen K, Seuberlich T, Dawson KLD, Zientara S, Laabassi F.Viral diseases cause significant economic losses within the equine population. Horses are susceptible to equine coronavirus (ECoV) and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), although only ECoV has been associated to clinical disease. The aim of this study was to investigate, for the first time in Algeria, the seroprevalence of ECoV and SARS-CoV-2 antibodies and the prevalence of ECoV infection in horses. In 2022, a total of 299 serum samples was collected from horses aged 1 to 27 years. Serological analysis for the presence of ECoV and SARS-CoV-2 was performed using a va...
Rimayanti R, Khairullah AR, Mustofa I, Utomo B, Lestari TD, Utama S, Akintunde AO, Mulyati S, Hernawati T, Dawood AQ, Riady G, Khan IU, Rasad SD.... is a protozoan parasite that causes the sexually transmitted disease known as "dourine" in horses. This chronic illness is directly spread from one animal to another during mating. Doflein proposed the name in 1901. Despite being distributed worldwide, the broad use of artificial insemination technology over the past three decades has resulted in only a few cases being documented. The condition is typically fatal and is characterized by gradual emaciation, nervous system involvement, and edematous lesions of the genitals. The incubation period between exposure and the onset of clinical sympt...
Clark KF, Lemcke RA, Gasiorowski JC, Wagner B.Lyme arthritis, a well-documented subtype of arthritis in humans and dogs, is caused by Borrelia burgdorferi and spread primarily through ticks (Ixodes spp.). While Lyme disease in horses is known to cause a variety of clinical signs, its involvement in the development of equine arthritis is controversial. As climate change enables tick populations to expand, more horses could be exposed to B. burgdorferi; therefore, a greater understanding of how Lyme disease affects joints is critical to preserving soundness. Objective: To determine if systemic, naturally occurring Lyme disease had a discern...
This study presents the first investigation of West Nile virus (WNV) seroprevalence among farmed horses in Kazakhstan. In 2024, a total of 368 serum samples were collected from horses across 106 settlements in 10 regions of the Republic of Kazakhstan. Using an enzyme-linked immunosorbent assay (ELISA), antibodies to WNV were detected in 32 horses (8.7%; 95% CI: 6.2-12.0%) from six regions. Among the seropositive animals, 26 (81.25%) were females and 6 (18.75%) were males, ranging in age from 1 to 19 years. No statistically significant association between sex and the presence of antibodies to W...
Costa VMD, de Aguiar Filho RN, Braz GF, Paz CFR, Drumond BP, Teixeira BM, Kroon EG.Equine infectious anemia (EIA) is a chronic disease caused by the equine infectious anemia virus (EIAV), a member of the Retroviridae family and the Lentivirus genus. Most infected animals are asymptomatic carriers, which makes clinical diagnosis challenging. This study aimed to conduct a clinical and laboratory evaluation of EIA in equids used for equestrian tourism in Jericoacoara National Park, Ceará, Brazil. A total of 112 equids (104 horses and 8 mules) underwent clinical, hematological, biochemical, serological, and molecular assessments. The EIAV seropositivity rates were 11.6% by AGID...
de Oliveira UV, Waap H, Gomes J, da Silva AN, Lacerda LC, da Silva Oliveira Costa T, de Sousa Lima T, Schares G, Pinheiro AM, Gondim LFP, Munhoz AD.The study aimed to assess the prevalence of horses reactive to Sarcocystis neurona, Neospora caninum, and Besnoitia besnoiti in horses from the state of Rio Grande do Sul, Brazil. Sera reactivity was tested by the Immunofluorescent Antibody Test (IFAT) in 354 equids sampled at slaughter, using a cut-off of 1:80 for S. neurona and 1:50 for N. caninum and B. besnoiti. Detection of N. caninum was attempted by PCR and bioassay in gerbils (Meriones unguiculatus) using brain samples collected from seropositive horses. Sera from 158 horses (44.63 %) (158/354; CI: 39.54-49.84 %) reacted to S. neuron...
Gonzálvez M, Franco JJ, Cano-Terriza D, Barbero-Moyano J, Jose-Cunilleras E, García J, Alguacil E, García-Bocanegra I.Equine Infectious Anemia (EIA) is a notifiable disease caused by Equine Infectious Anemia Virus (EIAV), with major sanitary and economic importance for equids worldwide. There is limited and outdated information on the circulation of EIAV in many European countries. In the present study, we aimed to assess virus exposure in different equid species in Western Europe. Methods: Between 2011 and 2023, a cross-sectional study was conducted to investigate the seroprevalence of EIAV in 1676 equids (1444 horses, 106 donkeys and 126 mules/hinnies) from four European regions: Andalusia (Southern Spain; ...
Rask E, Righetti F, Ruiz A, Bjerketorp J, Frosth S, Frykberg L, Jacobsson K, Guss B, Flock JI, Henriques-Normark B, Hartman E, Gustafsson A....Infection of susceptible horses with subspecies (), the causative agent of strangles, is associated with commingling. Exposure may occur among horses at equestrian events, sales, or horses moved among different equine stabling environments. Strangles can affect all horses on a farm, leading to the death of up to 10% of cases depending on their immunity status at the time of infection, the development of complications, the success of biosecurity measures, and the use of vaccination. The current retrospective study used ELISAs to measure the exposure of horses to at a farm that experienced an...
Cyathostomins and Anoplocephala perfoliata infect equids worldwide and both are of concern due to their potential to cause clinical disease. Although coprological techniques are commonly used for assessing helminth egg shedding these do not provide information about an individual's total parasite burden. Antibody-based tests provide information on infection levels within individuals. Objective: This study aimed to evaluate cyathostomin and Anoplocephala perfoliata occurrence in Italian horses using coprological and serum-based antibody detection methods and to analyse risk factors for infectio...
Mackowski M, Kajdasz A, Laskowska K, Cieslak J.Although serological and genetic studies of equine blood group systems have been conducted for many years, the molecular basis of erythrocyte antigens' variability has remained largely unexplored. In this study, we aimed to elucidate the genetic basis of serological variation within equine blood group K. Using mRNA extracted from peripheral blood samples (n = 100) collected from horses with known serological blood types (Ka or K-), we performed a transcriptome-wide association study (TWAS), which revealed a significantly associated region on equine chromosome 2 (ECA2). A detailed analysis ...
Beniwal N, Lal B, Mithina S, Verma CK, Kumar S, Phagna V, Jakhar K, Sonar S, Gupta V, Singh R, Kumar N, Tan CW, Thachamvally R, Singha H, Murzello K....Rabies is a neglected tropical zoonotic disease caused by rabies-virus (RV) infection and is responsible for almost 60,000 annual deaths globally, largely affecting the socio-economically disadvantaged population. Although fatality is preventable by immunization either before or after exposure with therapeutic antibodies, the high cost of prophylaxis or treatment limits their accessibility for the affected population. However, due to the almost 100% fatality rate in symptomatic individuals, almost 29 million annual vaccinations are performed, imposing high financial burden. Human transmission ...
Yang Y, Guo K, Xu L, Guo W, Dong M, Liu W, Li S, Zhang Z, Chu X, Wang Y, Zhang Z, Hu Z, Wang X.Equine influenza (EI), caused by the equine influenza virus (EIV), is an acute respiratory disease that has become enzootic worldwide, resulting in frequent outbreaks and substantial economic losses within the equine industry. In this study, we developed a competitive enzyme-linked immunosorbent assay (NP-cELISA) for the detection of antibodies against the EIV nucleoprotein (NP). The assay was designed by coating plates with purified monoclonal antibodies (mAbs) against the NP protein, followed by simultaneous incubation of the test serum samples and HRP-NP antigen in a competitive binding rea...
Kronenberg PA, Fouché N, Sekiya M, Liechti P, Frey CF, Mulcahy G, Eichenberger RM.Fasciola hepatica is a common trematode parasite of livestock in many regions, causing significant economic losses and affecting animal welfare. Horses rarely develop patent liver fluke infection. However, liver damage can affect animal health and welfare. Therefore, F. hepatica infection in horses may be underreported. Recently, a serological test for the antibody detection has been reported based on recombinant parasite cathepsin L1 (FhCL1) protease. Here, we optimized this enzyme-linked immunosorbent assay (ELISA) for antibody-detection in horses with equine fasciolosis based on recombinant...
Pettersson J, Levanov L, Tervo S, Hautala K, Aaltonen K, Utriainen M, Kareinen L, Gadd T, Sironen T, Vapalahti O, Kinnunen PM.Parapoxviruses (PPV) cause skin and mucous membrane signs to several animal species and humans worldwide. Equine parapoxvirus (EqPPV) was first detected in a sick horse in Finland in 2013. It is potentially zoonotic, and a similar virus has been detected in humans in the USA. In winter 2021–2022, EqPPV caused a large-scale pastern dermatitis epidemic in racehorses all over Finland. Field reports suggest that similar epidemics of unverified cause have also occurred in 2015 and 2019. The aim of this study was to develop a serological test and study the immune response, seroprevalence, and hist...
Zhang Y, Zheng J, Zhang H, Lin Y, Wang Y, Ma Z, Wei J, Zhou B, Zhong D.Getah virus (GETV), Japanese encephalitis virus (JEV), West Nile virus (WNV), and African horse sickness virus (AHSV) are mosquito-borne viruses threatening the health of racehorses. However, the systematic surveillance of these viruses among Shanghai racehorses remains lacking. Therefore, molecular and serological surveillance was conducted for these viruses in racehorses and mosquitoes at horse farms in Shanghai, China, during 2022 to assess their prevalence. Among 11,140 mosquitoes collected from seven farms across four districts, and were identified as the dominant species. RT-qPCR detec...
Mota RA, Silito IS, Martins MP, Oliveira PRF, Melo RPB, Vieira da Silva V, Jorge FR, Martins TF, Labruna MB.Tick-borne diseases are among the most widespread vector-borne infections, including zoonoses caused by bacterial and protozoal microorganisms. This study consisted of a serosurvey for Rickettsia spp. in 270 horses from 11 farms in the coastal region of Pernambuco, Brazil. Through the indirect immunofluorescence assay (IFA) based on antigens of five Rickettsia species, 164 (60.7%) horses were seroreactive to Rickettsia spp., with seroreactivity among farms ranging from 10 to 100%. Through molecular analysis, all 270 horse blood DNA samples tested negative by PCR assays targeting Anaplasmatacea...
Djellouli M, Eddaikra N, Beneldjouzi A, Benikhlef R, Ghimire R, Wilkins M, Lafri I.Cutaneous leishmaniosis remains a major public health concern in Algeria, predominantly affecting humans and dogs. However, the role of other animal hosts, particularly equids, in the transmission cycle remains underexplored. This cross-sectional study presents the first systematic investigation of Leishmania spp. exposure and infection in equids, along with associated environmental and host factors, from southeastern Algeria, El Oued Wilaya, the third most affected region for human cutaneous leishmaniasis in the country. A total of 122 equids from eight communes were clinically examined and t...
