Serodiagnosis in horses involves the detection and measurement of antibodies or antigens in the blood to diagnose infectious diseases and monitor immune responses. This diagnostic approach is based on serological tests that identify the presence of specific immune markers, providing insight into the horse's exposure to pathogens or vaccination status. Common serological tests used in equine medicine include enzyme-linked immunosorbent assays (ELISA), complement fixation tests, and virus neutralization tests. These tests are valuable for diagnosing conditions such as equine infectious anemia, strangles, and equine herpesvirus. This page compiles peer-reviewed research studies and scholarly articles that explore the methodologies, applications, and advancements in serodiagnostic techniques for equine health assessment.
House C, Mikiciuk PE, Berninger ML.Five serological methods of diagnosing African horse sickness were evaluated, using a battery of serum samples from experimental horses vaccinated and challenged with each serotype of African horse sickness virus (AHSV1 through AHSV9): agar gel immunodiffusion (AGID), indirect fluorescent antibody (IFA), complement fixation (CF), virus neutralization (VN), and enzyme-linked immunosorbent assay (ELISA). The 5 tests were also compared using a panel of field samples, convalescent equine sera with antibodies to domestic equine viral diseases, and sera from horses awaiting export. The ELISA describ...
Magnarelli LA, Anderson JF.Class-specific and polyvalent ELISA were developed to detect IgM antibody or total immunoglobulins to Borrelia burgdorferi in equine sera. Analyses of 122 serum specimens, collected during 1985 from horses and ponies in tick-infested areas of Connecticut, revealed IgM antibody in 41 (34%) samples; titration end points ranged from 1:160 to 1:2,560. In polyvalent ELISA, 73 (16%) of 454 serum specimens contained IgM and/or IgG antibody. Seropositivity was highest (32%) for blood samples collected during May. Both ELISA procedures had comparable sensitivities.
Palmer JE, Benson CE, Lotz GW.Ten healthy, mature ponies were orally infected with Ehrlichia risticii using Ehrlichia-infected P388D1 mouse monocyte tissue culture cells. Seven developed signs of equine ehrlichial colitis including fever, depression, anorexia, reduced borborygmi, increased abdominal hyperresonance, and diarrhoea. Three remained clinically normal apart from early fever in one. Indirect fluorescent antibody titres were detected in the clinically affected ponies by Days 12 to 17 post infection and increased rapidly to high levels (1:640 to 1:5120) which were maintained until the end of the observation period ...
Rossmanith W, Horvath E.After electrophoretic separation in SDS-PAGE structural proteins of the virus of Equine Infectious Anemia (EIA) were easily blotted by the semi-dry-blotting method onto nitrocellulose filters. Strips of these filters were used for antibody demonstration, and positive reactions thereof were intensified by a biotin-avidin-peroxidase system. Sensitivity of this system was so high as to allow readable interpretation of bands up to the dilution of 1:6,400 of a strongly positive serum. Frequently this procedure allowed to make a firm diagnostic Western-Blot diagnosis on far weaker equine sera. Inter...
Shankarappa B, Dutta SK, Sanusi J, Mattingly BL.Competitive enzyme-linked immunosorbent assay (CELISA), mediated by a monoclonal antibody designated HybI, was developed for the diagnosis of equine monocytic ehrlichiosis. Inhibition of binding of HybI by the horse antibodies to Ehrlichia risticii was optimum at dilutions of 1:20 for serum and 1:10,000 for HybI. Mean optical densities (ODs) of positive and negative sera were 0.158 and 0.855, respectively. A comparison of ODs obtained by CELISA and indirect enzyme-linked immunosorbent assay (ELISA) indicated a marked tendency of positive and negative samples to cluster separately with respect ...
Hall RF, Pursell AR, Cole JR, Youmans BC.An epizootic of equine infectious anemia (EIA) involved 35 horses on a farm in south Georgia. During a 126-day period, 21 of these horses became seropositive for EIA. After the initial diagnosis in July, the horses were tested every 7 to 10 days. At least one additional horse was found to be seropositive on each testing day. As soon as they were determined to be seropositive, the horses were removed from the herd and sent to slaughter. The removal of the seropositive horses, however, did not stop the epizootic. We believe the initial infection was from a 7-year-old stallion that recently had b...
Eguchi M, Kuniyasu C, Kishima M.The passive hemagglutination (PHA) test was improved to enable the detection of antibodies to Taylorella (Haemophilus) equigenitalis in the sera of mares. Horse red blood cells (RBC) fixed with glutaraldehyde were compared with similarly treated RBC of a cow, pig and sheep for the PHA test. The horse RBC were superior to those of the other animals tested in detecting mares affected with contagious equine metritis (CEM). A PHA test using these cells as indicator and an antigen prepared from T. equigenitalis by sonication following treatment with hyaluronidase was the most satisfactory in terms ...
Williamson CC, Stoltsz WH, Mattheus A, Schiele GJ.The complement fixation test (CFT), indirect fluorescent antibody test (IFAT), card agglutination test for trypanosomiasis (CATT) and enzyme-linked immunosorbent assay (ELISA) were compared in their application to the serological diagnosis of Trypanosoma equiperdum infection in 43 horses. The CFT remains a reliable test for dourine, especially in countries where other members of the subgenus Trypanozoon do not occur. The IFAT is a good 'back-up' test, but, requiring skilled operators it has the disadvantage of making it labour intensive, and interpretation of results subjective. This makes it ...
Tenter AM, Otte MJ, Gonzalez CA, Abuabara Y.Eighty-two equine sera from 13 farms in northern Colombia were examined for antibodies to Babesia caballi and B. equi using the complement fixation (CF) and the indirect fluorescent antibody (IFA) test. Seroreactors to both piroplasms were present on all farms. The IFA test indicated a prevalence of 90% for B. caballi and 94% for B. equi. The CF test detected antibodies to B. caballi in 41% and to B. equi in 65% of the animals. The prevalence of seroreactors in different age groups revealed a significant decline in CF antibodies to B. caballi in animals older than three years. IFA titres for b...
De Waal DT, Van Heerden J, Van den Berg SS, Stegmann GF, Potgieter FT.Both Babesia equi and Babesia caballi are endemic in large parts of South Africa. Attempts were made to obtain pure local isolates of both B. equi and B. caballi for the purpose of developing serological tests to study the epidemiology of equine babesiosis in this country. The indirect fluorescent antibody test was used to screen horses for B. equi and B. caballi in an endemic area. Seven horses and 3 donkeys between 3 and 36 months of age that tested negative were subsequently splenectomized. The splenectomy operation was performed through the abdominal approach. A 100% survival rate was achi...
Wilkins CA, Bergh N.An examination of the sera of 329 horses for L. pneumophila antibodies revealed a much lower exposure rate than that reported in the United States of America. Further serological investigations of persons closely associated with a sero-positive horse indicated that the horse could not be considered to be a source of infection but that both humans and animals were probably exposed to a common source of infection. The results showed that 192/329 (58.4%) of the sera tested negative, 114/329 (34.7%) had end-point titres of 1/2, 22/329 (6.7%) end-points of 1/16 and one an end point of 1/256 (0.3%)....
Sillerud CL, Bey RF, Ball M, Bistner SI.After the observation of 2 horses with uveitis on a horse farm in the Minnesota River valley, 100 horses from this geographic area were given ophthalmologic examinations and were evaluated serologically for leptospirosis. A statistically significant (P less than 0.001) association was observed between the finding of antibodies against Leptospira interrogans serovar pomona and uveitis.
Woolcock JB, Mutimer MD, Bowles PM.Normal horses of all ages regularly show evidence of having responded immunologically to R. equi, thus adding serological support to epidemiological evidence that this organism is a normal intestinal inhabitant. More animals from "diseased" farms show a stronger antibody response when compared with foals from "healthy" farms. Various serological tests have been used to detect evidence of infection and to relate antibody level to severity of disease. Anti-R. equi IgG antibody levels, as measured by ELISA, are raised significantly during natural infection. Clinical severity of pneumonia can be c...
Skalka B.The occurrence of equi-factor antibodies in sera of mares and their foals was studied on two horse breeding farms, one of which (Farm A) had a positive and the other (farm B) a negative history of R. equi infection of foals. The equi-factor neutralization (EFN) and the reverse Elek-Ouchterlony (REO) precipitation were used as assays. On Farm A, 25 mares positive in both tests (EFN+ REO+) and 25 mares negative in both tests (EFN- REO-) was chosen. On Farm B, a group of 25 EFN- REO+ mares and a group of 25 EFN- REO- mares were studied. The first serum samplings in mares were 1 week ante partum a...
De Lange JF, Gummow B, Turner GV, Redman AR.This is the first known isolation in the Republic of South Africa (RSA) of the serovar pomona from the organs of porcine foetuses as well as from the renal lymph nodes of slaughter pigs showing chronic nephritis. In addition, the serovar pomona was isolated from the kidneys of 87.5% of the slaughter pigs examined. The success of these isolations was attributed in part to the refining of 2 existing isolation techniques which complement each other. Using the microscopic agglutination test, serum samples taken from the same farming unit showed evidence of antibodies to the serovar pomona in 89 ou...
Gummow B, Herr S, Brett OL.A microtitration serum agglutination test, based on that used for brucellosis, has been developed to detect antibodies in the sera of horses exposed to the contagious equine metritis (CEM) organism. Two known positive sera were tested 100 times in 15 separate tests. The results were reproducible to within a twofold range. The test is capable of being carried out within 100 min.
Matthews AG, Waitkins SA, Palmer MF.The prevalence of antibody titres to a range of 20 leptospira antigens in the serum of horses and ponies with no ophthalmic abnormalities and with ophthalmoscopic evidence of endogenous uveal inflammatory disease was determined using a microscopic agglutination technique. Titres against leptospira antigens were observed in 13 out of 138 (9.1 per cent) animals with no ophthalmic abnormalities, and in three out of 27 (11.1 per cent) animals with anterior uveitis. Serovar sejroe was common to all seropositive animals with anterior uveitis. The results show that leptospira infection is not a major...
Calisher CH, Monath TP, Sabattini MS, Mitchell CJ, Lazuick JS, Tesh RB, Cropp CB.In 1983, 17 virus strains were isolated from mosquitoes collected during an outbreak of western equine encephalitis in Santa Fe Province, Argentina. Strains of western equine encephalitis, Venezuelan equine encephalitis, St. Louis encephalitis, and Antequera viruses were isolated, as were several bunyaviruses of the California and Bunyamwera serogroups and a new vesiculovirus. Complement fixation and neutralization tests were used to identify the California serogroup virus as a subtype of Melao virus, the Bunyamwera serogroup virus as a subtype of both Maguari and Playas viruses, and the vesic...
Gummow B, Herr S, Brett OL.A complement fixation test, using round-bottomed microtitration plates and an 8 channel microdiluter, based on that used for brucellosis by Herr, Huchzermeyer, Te Brugge, Williamson, Roos & Schiele, 1985, has been developed for use on the sera of horses to detect antibodies to the contagious equine metritis organism. The results with 2 known positive sera tested 116 times in 27 separate tests were reproducible for the most part within a twofold range. They seldom exceeded these limits and never exceeded a fourfold range. The test itself is capable of being carried out within 90 min. The test w...
Williamson CC, Herr S.The distribution of positive dourine cases found on the complement fixation test at the Veterinary Research Institute, Onderstepoort from 1981 to 1984, is recorded. Within the Republic of South Africa, foci of infection occurred in the Johannesburg, Pretoria, Potchefstroom, Rustenburg, Upington, Lichtenburg, Kroonstad, Louis Trichardt, Middelburg (Cape) and Mossel Bay state veterinary districts. In Bophuthatswana, Transkei, Lesotho, South West Africa and Swaziland, positive cases were also recorded. Anti-complementary activity of horse sera does not present a problem. In donkey and mule sera, ...
Ristic M, Holland CJ, Dawson JE, Sessions J, Palmer J.The recent establishment of a system for the continuous in vitro propagation of Ehrlichia risticii, the causative agent of equine monocytic ehrlichiosis (EME; synonym, Potomac horse fever), has facilitated the development of an indirect fluorescent antibody test for the diagnosis of this disease under laboratory and field conditions. The field diagnostic application of the test has aided in the recognition of the disease in 16 states of the United States and in 1 province of Canada. A limited epidemiologic study conducted between January and September 1985, in an area where the disease is know...
Calisher CH, Mahmud MI, el-Kafrawi AO, Emerson JK, Muth DJ.Paired sera from 28 nonvaccinated horses with serologically confirmed western equine encephalitis (WEE) virus infections were evaluated for immunoglobulin (Ig)M and IgG directed against WEE virus, by use of enzyme immunoassay. Twenty-one of the horses developed greater than or equal to 4-fold increases or decreases in serum IgM titers in paired serum samples, confirming the diagnosis of WEE in these horses. Of the remaining 7 horses, 1 had stable IgM titers, 1 had a 2-fold increase in IgM titer between paired sera, 2 had 2-fold decreases in IgM titer, and for 3 horses adequate volumes were not...
Mendoza L, Kaufman L, Standard PG.A practical, sensitive, and specific immunodiffusion test was developed for diagnosing and monitoring pythiosis in horses. Culture filtrates, a soluble cell mass, and trypsinized Pythium sp. antigens were evaluated against prepared rabbit anti-Pythium sp. serum and pythiosis horse case sera. The culture filtrate antigens demonstrated the greatest capacity for detecting precipitins and the greatest stability during storage. In contrast, the trypsinized antigens had the weakest capability for detecting multiple precipitins and the poorest stability. The 13 sera from horses with proven active pyt...
Tenter AM, Friedhoff KT.The sensitivity and specificity of the complement fixation (CF) test for the diagnosis of Babesia infections in equines was assessed, using the indirect fluorescent antibody (IFA) test as a reference. Antibodies were first detected between 11 and 20 days post infection (dpi) in the CF test and between 7 and 14 dpi in the IFA test in ponies infected experimentally with B. equi (USDA strain). The CF test became negative in four of five ponies 63-174 dpi although B. equi was demonstrated microscopically in two of these four ponies up to 364 and 455 dpi. The IFA test remained positive up to 476 dp...
Weiland G.The increasing horse trade requires a reliable immunodiagnosis of equine piroplasma infections due to import restrictions imposed by various countries, including the United States of America. It was the aim of our investigations to establish the suitability of serological tests for the detection of parasite carriers and, eventually, to differentiate between Babesia caballi and B. equi infections. The investigations were carried out on 11 ponies with experimentally-induced B. caballi and/or B. equi infection. The infections were confirmed by the demonstration of parasites in blood smears 2-13 d...
Sánchez-Matamoros A, Beck C, Kukielka D, Lecollinet S, Blaise-Boisseau S, Garnier A, Rueda P, Zientara S, Sánchez-Vizcaíno JM.African horse sickness (AHS) is a viral disease that causes high morbidity and mortality rates in susceptible Equidae and therefore significant economic losses. More rapid, sensitive and specific assays are required by diagnostic laboratories to support effective surveillance programmes. A novel microsphere-based immunoassay (Luminex assay) in which beads are coated with recombinant AHS virus (AHSV) structural protein 7 (VP7) has been developed for serological detection of antibodies against VP7 of any AHSV serotype. The performance of this assay was compared with that of a commercial enzyme-l...
Chien RC, Mingqun L, Yan Q, Randolph N, Huang W, Wellman M, Toribio R, Rikihisa Y.The tick-borne obligatory intracellular bacterium Anaplasma phagocytophilum infects humans as well as domesticated and wild animals, causing a febrile disease collectively called granulocytic anaplasmosis. The epidemiology and the host species specificity and zoonotic potential of A. phagocytophilum strains remain unclear. In this study, ankA (encoding ankyrin A) and p44 gene sequences of A. phagocytophilum were determined in clinical specimens from horses in Ohio and compared with those found in A. phagocytophilum strains from various hosts and geographic regions. With increasing numbers of s...
Baghdadi HBA, Abdelsalam M, Attia MM.Strongylus vulgaris, a devastating parasitic nematode in equids, causes life-threatening verminous aneurysms that are challenging to diagnose early. This study pioneered integrating nanotechnology into an indirect enzyme-linked immunosorbent assay (i-ELISA) system to enhance the sensitivity and specificity for detecting S. vulgaris larval antigens in equine serum samples, with PCR confirmation of the species. A conventional i-ELISA and an innovative nano-based ELISA were developed using excretory-secretory antigens from adult S. vulgaris worms. The nano-ELISA incorporated gold nanoparticles (1...
Hébert L, Froger D, Madeline A, Lecouturier F, Lemans C, Zientara S.Dourine is a sexually transmitted parasitic disease affecting equids. Its causative agent is referred to as and the prescribed serodiagnosis method is the complement fixation test (CFT). In the context of our European Reference Laboratory mandate for equine diseases (excluding African horse sickness), we organised dourine CFT inter-laboratory proficiency tests (ILPTs) in 2015, 2018 and 2022 to evaluate the performance of the European Union network of National Reference Laboratories (NRLs) for dourine. ILPT panels were composed of horse sera with or without antibodies against spp. originating...
Luciani M, Armillotta G, Di Febo T, Krasteva I, Ulisse S, Di Pancrazio C, Laguardia C, Perletta F, Serroni A, Maggetti M, Testa L, Sacchini F....The significance of as the causative agent of dourine cannot be understated, especially given its high mortality rate among equids. International movement of equids should be subject to thorough health checks and screenings to ensure that animals are not infected with . This involves the implementation of quarantine protocols, testing procedures, and the issuance of health certificates to certify the health status of the animals. Three proteins, the peptidyl-prolyl cis-trans isomerase (A0A1G4I8N3), the GrpE protein homolog (A0A1G4I464) and the transport protein particle (TRAPP) component, put...
Boch J.Babesia infections serologically diagnosed in horses, cattle and dogs in Southern Germany during the last few years are described. 321 sera of horses were examined for specific antibodies to Babesia by means of CFT and IIF in 1984; 18 sera reacted to Babesia equi and 4 to Babesia caballi antigen. In a cattle breeding area in the Western Allgäu 13% of 1616 cattle reacted positive to Babesia divergens antigen using IIF and ELISA; during the grazing season 1982 new latent infections were observed in 25 of 266 calves and heifers. Cases of introduced canine babesiosis are more frequent; 10 of 34 s...
Dzierzecka M, Kita J.In this investigation the Western blot test was treated as a method verifying results of the IFA, commercial ELISA and standardized ELISA tests (described in Part I). The verifying investigations were performed on 82 serum samples, which in the commercial ELISA were positive in 36 cases, dubious in 31 cases and negative in 15 cases as well as on 5 serum samples obtained from horses infected with Leptospira spp., which in the ELISA commercial were dubious (total of 87 sera samples). The antigens, against which the immunological response in horses was directed, were also established. The Milenia...
Hagiwara K, Momiyama N, Taniyama H, Nakaya T, Tsunoda N, Ishihara C, Ikuta K.Sero- and molecular-epidemiological studies on Borna disease virus (BDV) infection show that BDV RNA is not always detected in the peripheral blood mononuclear cells (PBMCs) from serum anti-BDV antibody-positive individuals such as horses, sheep, cattle, cats, and humans. In this study we demonstrated BDV RNA signals by polymerase chain reaction only in restricted regions of the brain from horses with locomotor disease. Four of six horses examined showed apparently positive reactions for anti-BDV antibodies. Specific regions of the brain of these four horses were positive for BDV RNA but the i...
Williamson CC, Stoltsz WH, Mattheus A, Schiele GJ.The complement fixation test (CFT), indirect fluorescent antibody test (IFAT), card agglutination test for trypanosomiasis (CATT) and enzyme-linked immunosorbent assay (ELISA) were compared in their application to the serological diagnosis of Trypanosoma equiperdum infection in 43 horses. The CFT remains a reliable test for dourine, especially in countries where other members of the subgenus Trypanozoon do not occur. The IFAT is a good 'back-up' test, but, requiring skilled operators it has the disadvantage of making it labour intensive, and interpretation of results subjective. This makes it ...
Bernard WV, Cohen D, Bosler E, Zamos D.Blood samples obtained from 13 of 100 (13%) and 6 of 91 (7%) horses at the George D. Widener Hospital for Large Animals in the months of June and October, respectively, had antibody to Borrelia burgdorferi as determined by ELISA. Horses from the states of New York, Maryland, Delaware, New Jersey, and Pennsylvania were seropositive for B burgdorferi. The frequency of antibody response in horses from New Jersey was greater (P less than 0.05) than the frequency of antibody response in horses from Pennsylvania or that of horses from the other states combined. Statistically significant difference w...
Hobo S, Niwa H, Anzai T.The reactivity of synthesised peptide sets for the M-like proteins SeM and SzPSe with sera from horses infected with Streptococcus equi or Streptococcus zooepidemicus, or control horses, was investigated by an ELISA. Seventeen horses were infected experimentally with S equi or S zooepidemicus, convalescent sera were obtained from 25 horses and control sera were obtained from 1945 horses. The serum antibody responses of individual horses to the peptide sets were highly variable. Some of the peptide sets for SeM reacted strongly with the sera from the horses infected experimentally with S equi, ...
De Lange JF, Gummow B, Turner GV, Redman AR.This is the first known isolation in the Republic of South Africa (RSA) of the serovar pomona from the organs of porcine foetuses as well as from the renal lymph nodes of slaughter pigs showing chronic nephritis. In addition, the serovar pomona was isolated from the kidneys of 87.5% of the slaughter pigs examined. The success of these isolations was attributed in part to the refining of 2 existing isolation techniques which complement each other. Using the microscopic agglutination test, serum samples taken from the same farming unit showed evidence of antibodies to the serovar pomona in 89 ou...
Ghram A, Chabchoub A, Turki I, Boussetta M, Ibn Amor H, Ghorbel A.A seroepidemiological survey was realized in the Nord-Est Tunisia to study the prevalence of complement fixing and neutralizing antibodies to equine rhinopneumonitis and viral arteritis of horse, respectively. Four hundred sera were tested, using complement fixation reaction and seroneutralization test. The results show that 8.75% of sera have antibodies to viral arteritis and only 1.25% are positive for equine rhinopneumonitis.
Hall RF, Pursell AR, Cole JR, Youmans BC.An epizootic of equine infectious anemia (EIA) involved 35 horses on a farm in south Georgia. During a 126-day period, 21 of these horses became seropositive for EIA. After the initial diagnosis in July, the horses were tested every 7 to 10 days. At least one additional horse was found to be seropositive on each testing day. As soon as they were determined to be seropositive, the horses were removed from the herd and sent to slaughter. The removal of the seropositive horses, however, did not stop the epizootic. We believe the initial infection was from a 7-year-old stallion that recently had b...
Ali HHM, Al-Bayati LH. is one of the most important pathogens, which causes a marked economic loss in small ruminants, in particular sheep, worldwide. This study aimed to detect the prevalence of in the sera of aborted ewes in Wasit province, Iraq, using the enzyme-linked immunosorbent assay (ELISA), followed by the investigation of the main histopathological alterations that occurred in some organs of the dead newborns. Out of 180 tested samples by ELISA, 32.22% of the evaluated animals showed positive reactions to IgG antibodies toward . Concerning the titers of the infection of seropositive ewes, there were sig...
Böse R, Peymann B, Barbosa IP.Sera from 60 horses held in breeding herd in Brazil were examined monthly by ELISA, immunofluorescence antibody test (IFAT) and Western blot. All foals had maternal antibodies detectable by ELISA and IFAT, and sero-conversion took place between the 2nd and 5th month of age. The 48 and 50 kDa antigens were recognized first in the course of infection. Of 79 sera taken after sero-conversion 78 reacted with the 48 kDa antigen, 76 with the 50 kDa, 50 with the 70 kDa, 54 with the 112 kDa, 72 with the 141 kDa antigen. In general, sera from horses older than 1 year reacted with all 5 diagnostic antige...
Oliveira Filho RB, Campos Malta K, Assis Santana VL, Vance Harrop MH, Tancler Stipp D, Friguglietti Brandespim D, Aparecido Mota R....The present study, the first to spatially characterize Leptospira spp. infection among equids in the Brejo Paraibano micro-region of the Paraiba state in the northeast of Brazil, investigated 257 animals in 26 farms properties. Serum samples from 204 horses, 46 mules and seven donkeys were serologically diagnosed using the microscopic agglutination test (MAT). The distribution of Leptospira spp. was studied by employing specific antigens from 24 different Leptospira serovars. All farms were georeferenced and their distribution visualised on a map of the Brejo Paraibano micro-region. In additio...