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.
Herr S, Huchzermeyer HF, Te Brugge LA, Williamson CC, Roos JA, Schiele GJ.The same techniques may be used in the complement fixation test (CFT) for the serological diagnosis of bovine brucellosis, Johne's disease (paratuberculosis), dourine, equine piroplasmosis and Q fever (caused by Coxiella burnetii). The reproducibility of results is excellent, falling for the most part within the twofold range and never exceeding the fourfold range. Agreement with other laboratories is excellent (i.e. within twofold) in the case of brucellosis and equine piroplasmosis antibody titres. A good correlation between the occurrence of the disease and serological reactions is found on...
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...
Khan ZU, Misra VC, Randhawa HS.Prevalence of serum precipitins against Micropolyspora faeni, Thermoactinomyces vulgaris and Aspergillus fumigatus, employing the counterimmunoelectrophoresis (COIE) and Ouchterlony's double diffusion (DD) techniques, is reported in 162 of the equines stationed at two military installations in north-western India. M. faeni specific precipitins were demonstrable in 58 of 112 mules from site I in the mountainous region whereas the results were negative for all of the 50 horses examined from site II located in the plains. Of the 58 M. faeni positive mules, 45 (78%) had signs and symptoms suggesti...
Mettler NE, Fernández AS, Di Santo MI, Pardo DA.Sera from 282 equines from Tandil country and surroundings were investigated searching for hemagglutination inhibition (HI), Complement fixation (CF), and Neutralizing (NT) antibodies against three flavivirus:Ilheus, St. Louis Encephalitis, and Yellow Fever from the Togaviridae family. Sera were collected between 3-20-79 and 11-25-80 from 10 different places in Tandil and Ayacucho countries. Animals ranged from 45 days to 27 years old. Forty nine of them reacted with one or more flavivirus by HI and/or CF tes representing a prevalence of 17.4% for this antigenic complex. Twenty four of them ne...
Cho SN, Collins MT, Reif JS.The indirect fluorescent antibody test was used to examine 109 samples of equine sera randomly selected from serum pools. Results were compared with titers obtained by the microagglutination (MA) test. A high correlation (r = 0.89) was found between titers measured by the 2 tests. Blood samples were obtained serially from a total of 156 horses at a research farm and the sera were tested against Legionella pneumophila serogroups 1 through 4 using the MA test; 29 horses (19%) seroconverted to at least 1 serogroup of L pneumophila. The indirect fluorescent antibody test substantiated the results ...
Prescott JF, Coshan-Gauthier R, Barksdale L.Antibody to equi factor(s) in cases of Corynebacterium equi pneumonia in foals was detected using C. pseudotuberculosis exotoxin sensitized calf red blood cells. The test was standardized using antitoxin produced in rabbits by injection of equi factor(s). All sera from ten foals with culture-diagnosed C. equi pneumonia had antibodies to equi factor(s) (titre range 8-256, mean 74.0) and nine sera from 11 foals with suspected C. equi pneumonia also showed antibodies (titre range 4-512, mean 136.4). Two of five pneumonia foals with transtracheal aspirate cultures not yielding C. equi had such ant...
Ribbeck R, Ilchmann G, Hiepe T.Research into the immunological diagnosis of gasterophilosis. So far there have been no reliable methods of diagnosing equine gasterophilosis intra vitam. Horses from the G.D.R. and the M.P.R. spontaneously infected with Gasterophilus spp. were tested for antibodies by the immunotechniques of counterimmunoelectrophoresis after Pesendorfer, passive haemagglutination and the intradermal test using antigen made from larvae of all 6 Gasterophilus spp. present in the palaearctis. All 3 techniques produced positive results. The intradermal injection produced an immediate reaction. A correlation betw...
Obi TU, Taylor WP.Serum samples collected from 1,197 goats, 586 sheep, 254, cattle, 55 dogs and 44 horses were examined for antibodies to adenovirus by the agar-gel precipitation test. Results show that 17.7% of the goats, 18.4% of the sheep, 4.3% of the cattle, and 4.5% of the horses had precipitating antibodies. None of the dog sera examined was positive. The results seem to indicate a moderate level of previous exposure to adenovirus infection especially among goats and sheep in Nigeria.
Dolan M, Cargill C, Martin F, Davenport P, Franks D, Lightfoot J.A bacteriological and serological survey for evidence of contagious equine metritis (CEM) was made during the 1980 breeding season on 3 horse studs in South Australia with a history of previous infection. Swabs from the clitoral sinus and the cervix were cultured for Haemophilus equigenitalis and serum was screened for antibody using the complement fixation test (CFT) and the enzyme-linked immunosorbent assay (ELISA). The specificity of both tests was greater than 0.99 but the ELISA was more sensitive in detecting antibody in infected mares. On the evidence presented it was concluded that H. e...
Ellis WA, O'Brien JJ, Cassells JA, Montgomery J.Thirteen strains of pathogenic leptospires were isolated from 12 of 91 horses; seven strains belonged to the Australis serogroup (serotype bratislava) with three, two and one strains belonging to the Icterohaemorrhagiae, Hebdomadis (serotype hardjo) and Autumnalis serogroups respectively. Using leptospires isolated from horses and others representing the known parasitic Leptospira serogroups, a sample of 650 mares' sera was tested for agglutinating antibodies. Antibodies were found in 89.1 per cent of sera. The predominant reaction was to serotype bratislava, strain S/1334/79, isolated in this...
Calisher CH, Emerson JK, Muth DJ, Lazuick JS, Monath TP.Sera from horses and human beings with clinically diagnosed western equine encephalitis (WEE) virus infections were tested for hemagglutination-inhibition (HI), complement-fixation (CF), and neutralizing (N) antibody to WEE virus. These tests confirmed infection in 43.8% (HI), 56.3% (CF), and 80.4% (N) of horses and 54.5% (HI), 59.1% (CF), and 77.3% (N) of human beings. Use of the N test as an adjunct to the HI and CF tests increased the likelihood of serologic confirmation to 91.7%. In both horses and human beings, N antibody increased steeply at the end of the 1st week after onset. The resul...
Hathaway SC, Little TW, Stevens AE, Ellis WA, Morgan J.Eighteen isolates from the Australis serogroup from free-living and domestic animals were identified using the cross agglutination absorption test. Serovar muenchen was found only in England and Wales in wood mice, short tailed and bank voles, a grey squirrel and a pig. Serovar bratislava was found in hedgehogs in England, Wales and Northern Ireland and also in a brown rat from Northern Ireland. Serovar bratislava was isolated from sheep in both England and Northern Ireland and from horses in Northern Ireland. The distribution of these serovars in relation to possible maintenance hosts is disc...
Suzuki T, Ueda S, Samejima T.An enzyme-linked immunosorbent assay (ELISA) was elaborated for the detection of specific antibody to equine infectious anemia (EIA) antigen. Sera from horses experimentally infected with EIA virus were assayed by ELISA, complement fixation (CF) and immunodiffusion (ID) tests for antibody to EIA antigen. The ELISA technique was found to be much more sensitive than CF and ID tests. In addition, EIA specific antibody could be detected by ELISA at an earlier stage of infection than by CF or ID techniques. The applicability of the technique to diagnosis of EIA is discussed.
Kirchhoff H, Ammar AM, Heitmann J, Dubenkropp H, Schmidt R.Sera from horses with respiratory disease (RD) have been investigated using the complement fixation test, indirect hemagglutination test, enzyme immune assay, and the metabolic inhibition test, and sera from mares after abortion, using the complement fixation test, indirect hemagglutination test and enzyme immune assay, for antibodies against Mycoplasma equirhinis, M subdolum, M. equigenitalium, M. pulmonis, M. felis, Acholeplasma laidlawii, A. hippikon and A. equifetale. Antibodies were found against all mycoplasma and acholeplasma species tested, more often against acholeplasmas. The antibod...
Collins MT, Cho SN, Reif JS.We examined more than 2,800 human and animal sera for antibodies to four serogroups of Legionella pneumophila by using the microagglutination test. Antibody titers of greater than or equal to 1:64 were considered positive. The occurrence of positive equine sera (31.4%) was significantly higher than the occurrence of positive sera in cattle (5.1%), swine (2.9%), sheep (1.9%), dogs (1.9%), goats (0.5%), wildlife (0%), and humans (0.4%). The highest titer measured in horses was 1:512. The occurrence of positive sera in horses was related directly to age. In horses less than or equal to 1, 2 to 3,...
Caporale VP, Biancifiori F, Frescura F, Di Matteo A, Nannini D, Urbani G.Comparative tests such as FAT, ELISA, RIA, IEO and CF in the diagnosis of dourine in the horse have proved a satisfactory concordance ratio of the ELISA with CF, which seems to be the most reliable test. Discrepancies have been observed as to the other tests which appear less sensitive than CF test.
Schönberg A, Caldas EM, Sampaio MB, Costa E, Plank SJ.IN the State of Bahia (Brazil) the leptospirin produced in Germany for experimental use by the Institute for Veterinary Medicine, Federal Health Office, Berlin, was administered to humans and animals in order to diagnose leptospirosis in collaboration with this Institute. The results were compared with the microscopic agglutination reaction. The total number of test persons or animals was 268; this group included 81 human patients. 60 heads of cattle, 50 goats, 40 pigs, 25 horses, and 12 dogs. All were tested serologically and simultaneously the intracutaneous test was carried out. This test w...
Higgins R, Cayouette P, Hoquet F, De LaSalle F.During a period of 30 months, from January 1977 to June 1979, Leptospira agglutinins were detected in 355 (6%) of 5841 bovine sera, 52 (10.1%) of 511 porcine sera, one (5%) of 20 equine sera and one (12.5%) of eight canine sera. Bovine, porcine and equine sera reacted predominantly with L. pomona. Reactors to L. hardjo/sejroe, L. icterohaemorrhagiae and L. grippotyphosa were also detected in cattle. One porcine serum reacted with L. grippotyphosa and one canine serum with L. icterohaemorrhagiae. Al the sera originated from suspected cases of leptospirosis.
Timoney PJ.The usefulness of the double immunodiffusion test for the diagnosis of louping ill infection was investigated. Whereas louping ill viral antigen was not detected in brain material from field cases of the infection, its presence was readily confirmed in suckling mouse brain isolates of the virus. The double immunodiffusion test was found to be unreliable as a serological test for the retrospective diagnosis of louping ill infection in the horse.
Sharma VD, Sethi MS, Yadav MP, Dube DC.Sero-prevalence of brucellosis in man and animals was studied during the years 1976 and 1977. Samples were collected from Hospitals/slaughter houses/livestock farms located in Delhi and different districts of Uttar Pradesh (U.P.). The sera samples tested were from 1685 men, 1607 goats, 438 sheep, 244 pigs, 361 cattle, 551 buffalos, 50 dogs, 318 equine and 43 free living animals. The percentage of seropositivity, excluding doubtful ones, was recorded as: man 0.89, goat 5.53, sheep 3.42, pigs 15.98, cattle 6.37 buffalo 4.9 and equine 12.89. Additionally an evidence of agglutinins was also detect...
Giles C, Cavanagh HM, Noble G, Vanniasinkam T.There are currently two known serotypes of equine adenovirus (EAdV), equine adenovirus type 1 (EAdV1) and equine adenovirus type 2 (EAdV2); EAdV1 is predominantly associated with upper respiratory tract infections while EAdV2 appears to have a higher association with gastrointestinal infection, however, very little is known about the prevalence of these viruses in horse populations in Australia. In this study we tested 122 serum samples obtained from horses in New South Wales, Australia, using a standard serum neutralization (SN) assay and ELISA. Ninety-seven of the 122 sera displayed had mode...
Sebek Z, Wallner H, Sixl W, Kaaserer G, Valová M.Results are presented of a serological examination of 1,547 domestic animals (cattle, pig, sheep, horse, goat, dog, cat) from 9 Tyrolian districts (Austria), performed in order to disclose the incidence of leptospirosis. Completely significant titres were domonstrated by means of the MAL test in the serotypes icterohaemorrhagiae or copenhageni, sorex-jalna, bratislava, sejroe and saxkoebing. In addition, antibodies were confirmed against L. bataviae, L. pomona, L. tarassovi and L. bulgarica, but the titres were insignificant. Of the animals examined, 7.2% gave positive reactions in titres of 1...
Cook AG, Buechner-Maxwell V, Morrow JK, Ward DL, Parker NA, Dascanio JJ, Ley WB, Cooper W.Horses that are exposed to Sarcocystis neurona, a causative agent of equine protozoal myeloencephalitis, produce antibodies that are detectable in serum by western blot (WB). A positive test is indicative of exposure to the organism. Positive tests in young horses can be complicated by the presence of maternal antibodies. Passive transfer of maternal antibodies to S. neurona from seropositive mares to their foals was evaluated. Foals were sampled at birth (presuckle), at 24h of age (postsuckle), and at monthly intervals. All foals sampled before suckling were seronegative. Thirty-three foals f...
Obi TU, Taylor WP.Serum samples collected from 1,197 goats, 586 sheep, 254, cattle, 55 dogs and 44 horses were examined for antibodies to adenovirus by the agar-gel precipitation test. Results show that 17.7% of the goats, 18.4% of the sheep, 4.3% of the cattle, and 4.5% of the horses had precipitating antibodies. None of the dog sera examined was positive. The results seem to indicate a moderate level of previous exposure to adenovirus infection especially among goats and sheep in Nigeria.
Štritof Z, Mitchell C, Turk N, Habuš J, Hađina S, Perharić M, Waller AS.Clinical cases resembling strangles are regularly seen in some areas of Croatia. However, there are no data on the prevalence of infection and the clinical forms or geographic distribution of the disease. The aim of this study was to determine the seroprevalence of Streptococcus equi subspecies equi in horses resident in Croatia, in order to estimate the geographic distribution of infection. The study included 291 horse sera from the eight counties where the majority of Croatian horses are kept. Sera were tested by indirect ELISA (iELISA) for the presence of serum antibodies against S. equi pr...
Wassall DA, Gregory RJ, Phipps LP.The detection of antibodies against Trypanosoma equiperdum in 689 equid sera was compared by enzyme-linked immunosorbent assay (ELISA), the complement fixation test (CFT) and an indirect immunofluorescent test (IIF). CFT was the least sensitive technique, susceptible to anti-complementary factors and the most technically demanding. IIF was more sensitive, but was only suitable for testing limited numbers of samples. In this study, ELISA was the most sensitive test, the least labour intensive and lends itself to a considerable degree of automation. It is suggested that ELISA would be relatively...
Kong XG, Pang H, Sugiura T, Sentsui H, Onodera T, Matsumoto Y, Akashi H.Equine infectious anemia virus (EIAV) core proteins were obtained from a baculovirus expression system. Recombinant baculoviruses (rBVs) highly expressed the Gag precursor and p26 antigens in an rBV-infected Sf21 cell culture supernatant. Enzyme-linked immunosorbent assay (ELISA) and agar gel immunodiffusion (AGID) were conducted using the expressed proteins to detect antibodies from experimentally infected horses. The expressed antigens showed low background levels, high specificity and sensitivity in ELISA and AGID. The results of the serological tests using the expressed antigens were ident...
Sengupta PP, Rudramurthy GR, Ligi M, Jacob SS, Rahman H, Roy P.Trypanosoma evansi, a haemoflagellated protozoan parasite, is responsible for chronic as well as the acute debilitating disease called surra in a wide range of herbivores and carnivores including domestic and wild animals. Since the parasite is having wide host range, there is a need for diagnostic test which can detect the T. evansi specific antibody in different species of animals for generating sero-surveillance data. In the present study we developed and evaluated competitive enzyme immunoassay using monoclonal antibodies (MAbs) raised against recombinant variable surface glycoprotein (rVS...
Farias IF, Souza EAR, Serpa MCA, Palha FS, Oliveira GMB, Labruna MB, Horta MC.Brazilian spotted fever (BSF) is a common tick-borne disease caused by Rickettsia rickettsii. Horses are the primary hosts of the main vector, Amblyomma sculptum, and are considered efficient sentinels for circulation of Rickettsia. Therefore, the aim of this study was to detect antibodies reactive to R. rickettsii antigens in horses from a non-endemic area in the north-central region of Bahia state, Brazil. Blood samples and ticks were collected from 70 horses from the municipalities of Senhor do Bonfim, Antônio Gonçalves, Pindobaçu, and Campo Formoso. The sera obtained were tested by an i...
Kumar S, Malhotra DV, Dhar S.The present study aimed to develop Dot-ELISA, complement fixation test (CFT) and capillary tube agglutination test (CAT) for serodiagnosis of Babesia equi infection and to compare their sensitivity with each other. For this study, sequential serum samples were collected from four donkeys experimentally infected with B. equi up to 90 days post infection (P.I.). B. equi antigen was prepared from the blood of a donkey showing more than 80% parasitaemia. Dot-ELISA, CF and CA tests were standardized as per the standard method. While performing CFT, it was observed that CFT standardized for the donk...
Zilch TJ, Lee JJ, Bressan GC, McDonough SP, Mohammed HO, Divers TJ, Chang YF.The current gold standard diagnostic test for leptospirosis is the microscopic agglutination test (MAT), which has many drawbacks; therefore, the development of a better and easier serological test for leptospirosis is needed. Objective: To apply reverse vaccinology (RV) and antigenic selection on the assortment of leptospiral targets and evaluate their potential for use as reagents for the diagnosis of equine leptospirosis. Methods: Cross-sectional study. Methods: The antigenic selection parameters were: proteins with antigenicity score ≥0.5 (VaxiJen), at least one B cell epitope and size b...
Milillo P, Traversa D, Elia G, Otranto D.Larvae of Rhinoestrus spp. (Diptera, Oestridae) infect nasal and sinus cavities of horses, causing a nasal myiasis characterized by severe respiratory distress. Presently, the diagnosis of horse nasal botfly relies on the observation of clinical signs, on the post mortem retrieval of larvae or on molecular assays performed using pharyngeal swabs. The present study was carried out to characterize larval somatic proteins and salivary glands of Rhinoestrus spp. in a preliminary assessment towards the immunodiagnosis of equine rhinoestrosis. Out of the 212 necropsied horses 13 were positive for th...
Bannai H, Tominari M, Kambayashi Y, Nemoto M, Tsujimura K, Ohta M.Antibody response in horses after accelerated-schedule Getah virus vaccination was evaluated for its potential adoption during outbreaks. One-year-old Thoroughbred horses received two doses of priming vaccinations following an accelerated schedule (accelerated group: 14-day interval, n = 30) or the conventional schedule (control group: 28-day interval, n = 30). At Day 14, both groups showed similar seropositive rates (66.7% in control group and 73.3% in accelerated group) and geometric mean (GM) virus-neutralizing titers (5.2 [95% confidence interval (CI), 3.0-8.8] in control group and 5.3...
Sengupta PP, Rudramurthy GR, Ligi M, Roy M, Balamurugan V, Krishnamoorthy P, Nagalingam M, Singh L, Rahman H.Trypanosoma evansi, a haemoflagellate, causes "surra" an important chronic wasting disease of a wide range of wild and domestic herbivorous and carnivorous animals including cattle, buffaloes, camels, horses, etc. The untreated recovered animal can act as a carrier without exhibiting the disease symptoms and can be a source of infection to healthy animals. The diagnosis and subsequent treatment of the carrier animals is helpful to curb the disease. As the parasitaemia in carrier animals is very scanty, the conventional blood smear examination, which is widely practiced in the field, cannot det...
Mazzei M, Savini G, Di Gennaro A, Macchioni F, Prati MC, Guzmàn LR, Tolari F.West Nile virus (WNV) is a mosquito-borne virus belonging to the family Flaviviridae included in the Japanese encephalitis antigenic complex (JEAC). A seroepidemiological study was carried out in 2011 using 160 horse sera collected from different areas of Bolivia to investigate the presence of WNV antibody. A high proportion (59.4%) of the tested sera were positive to a commercially available WNV competitive enzyme-linked immunosorbent assay (C-ELISA). Sixty-six randomly selected C-ELISA-positive sera were further tested by WNV plaque reduction neutralization test (PRNT), virus neutralization ...
Broeckl CV, Hiereth S, Straubinger RK.Diagnosis of equine Lyme borreliosis (LB), an infection caused by members of the Borrelia burgdorferi sensu lato complex (Bbsl), is challenging due to the nonspecific clinical signs of the disease and due to the variety of non-standardized serological tests. Specific vaccine-induced antibodies against LB, providing an effective protection against the infection, complicate the issue further. The standard for the detection of specific antibodies against Bbsl is a two-tier test system based on an enzyme-linked immunosorbent assay (ELISA) or indirect fluorescent antibody test (IFA) for antibody sc...
Punyadarsaniya D, Taesuji M, Rattanamas K, Ruenphet S.Standard serological tests post-vaccination, such as enzyme-linked immunosorbent assay (ELISA), complement fixation, and virus neutralization, are crucial for monitoring African horse sickness (AHS). However, the availability of commercial test kits such as blocking ELISA varies by regions; while they are commonly used in Africa and Europe, their limited availability and high cost in Thailand present significant challenges. Therefore, this study aimed to evaluate an alternative approach using an in-house indirect ELISA based on cell-based monovalent and polyvalent strains of live attenuated AH...
Schwarzer A, Ziegler U, Fertey J, Kreuz M, Vahlenkamp TW, Groschup MH, Ulbert S.West Nile virus (WNV), Usutu virus (USUV) and tick-borne encephalitis virus (TBEV) are worldwide endemic zoonotic orthoflaviviruses, often co-circulating in the same areas. Serological studies in animals, mostly birds and horses, are important means to monitor the spread of these viruses and the infection risks for humans. However, cross-reactive antibodies to these structurally similar flaviviruses frequently impact serological differentiation in enzyme-linked immunosorbent assays (ELISAs), hence time-consuming virus neutralization tests (VNTs) have to be employed in laboratories with high bi...
Ichikawa Y, Iinuma Y, Okagawa T, Shimbo R, Enkhtuul B, Khurtsbaatar O, Kinoshita Y, Niwa H, Aoshima K, Kobayashi A, Batbaatar V, Ohashi K, Kimura T.Glanders is a World Organization for Animal Health (WOAH)-notifiable equine disease caused by the infection of Burkholderia mallei, and is endemic in Mongolia, South Asia, Africa, and South America. While the complement fixation test (CFT) has been widely used for serodiagnosis of glanders and is considered a standard serological test, it has several limitations. These limitations include poor specificity, labor intensive techniques, variability in antigen and protocol. Consequently, indirect enzyme-linked immunosorbent assays (iELISAs) based on recombinant proteins have been developed as alte...
