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.
Gerdes GH, Pieterse LM.Virus was isolated from the blood of horses (n = 5) showing fever and jaundice and was identified as equine encephalosis virus. In cross neutralisation tests, the isolates were shown to belong to a new serotype related to Gamil, one of the 6 known serotypes of equine encephalosis virus. The name Potchefstroom has been proposed for this new serotype.
Fikrig E, Magnarelli LA, Chen M, Anderson JF, Flavell RA.Enzyme-linked immunosorbent assays (ELISA) and immunoblots using either whole-cell lysates of Borrelia burgdorferi or an antigenic region of flagellin (41-G) as the antigen were performed, and the abilities of the two assays to detect antibodies to this spirochete in dog, cottontail rabbit, and horse sera were compared. Assays using whole-cell B. burgdorferi lysates as the antigen were more sensitive for detecting antibodies. ELISA with 41-G as the antigen were specific for Borrelia antibodies but were not as sensitive as the assays with whole-cell lysates coated to the solid phase. Use of rec...
Escartin-Peña M, Bautista-Garfias CR.Sera from 41 horses and 159 donkeys, from twelve States of México, were tested to ascertain anti-Gasterophilus circulating antibodies by double immunodiffusion (DD), counterimmunoelectrophoresis (CIE), indirect haemagglutination (IH), thin layer immunoassay (TIA) and diffusion-in-gel ELISA (DIG-ELISA) methods using crude somatic antigen from third instar larvae of G. intestinalis (DeGeer). At necropsy, 33/41 horses and 24/159 donkeys were found to be parasitized by G. intestinalis and/or G. nasalis (L.). Gasterophilus intestinalis was the species most commonly found in the equines. Analysis o...
Afshar A, Shakarchi NH, Dulac GC.Two competitive (C) enzyme-linked immunosorbent assays (ELISAs) were developed for the detection of antibodies to vesicular stomatitis virus (VSV) in animal sera. The assays are based upon the availability of polyclonal antibodies (PAbs) from mouse ascitic fluids prepared against the New Jersey (NJ) and the Indiana (IN) VSV serotypes. The assays were performed by the immobilization of VSV-NJ and VSV-IN antigens on a solid phase (microtiter plate). Appropriately diluted test serum mixed with an equal volume of serotype-specific PAb was allowed to incubate in the presence of the relevant VSV ant...
Paweska JT, Barnard BJ.This paper reports the first serological evidence of exposure of donkeys to equine arteritis virus. Seven hundred and thirty-four serum samples collected between 1989 and 1992 from donkeys in different areas of South Africa were examined for the presence of antibodies against this virus by a microneutralization test. Seventeen percent of serum samples tested positive. The distribution of seropositive animals varied from none in the western Cape Province and the Transvaal Highveld to 30% in the northern Transvaal. The country-wide distribution of serologically positive donkeys suggests a longst...
Miyamoto C, Takashima I, Karaiwa H, Sugiura T, Kamada M, Hashimoto N.To investigate the overall prevalence of chlamydial infections in light (i.e. non-draught) horses in Japan, 599 sera obtained from 12 localities in 1991 were tested for complement fixation antibodies. The mean antibody positive rates of the all sera were 15.2% (91/599) and the regional positive rates were higher in Honshu (19.1%, 48/251) and Kyushu (20.0%, 20/100) than in Hokkaido (9.3%, 23/248). In Honshu, the highest rate (56.0%, 28/50) was observed in Utsunomiya. Analysis of the positive rate in different age groups showed that the 2-5 years age-group had the highest prevalence of chlamydia...
Aviles G, Bianchi TI, Daffner JF, Sabattini MS.It is shown here the WEE virus activity in ARgentina in 1983-1986 post-epizootic period. A surveillance system was established by the equine case notification and the sentinel animal method. Among the thirteen equine focus notified between September 1983-September 1985 in Córdoba and Santa Fe Provinces, 5 presumptive cases out of 16 sick horses were confirmed by the hemagglutination inhibition test for WEE epizootic virus. Twenty eight notified human cases were studied with negative results. The neutralizing antibody (Ac NT) prevalence among sentinel horses in Córdoba Province (4%) was lower...
Monzón CM.A direct agglutination test is described to diagnose 'Mal de Caderas' caused by Trypanosoma evansi. The antigen used was a suspension of trypsin-treated parasites stabilized with formalin. The test was evaluated in horses with both natural and experimental infections. Test sensitivity and specificity were 94 and 97%, respectively. Treatment of serum with 2-mercaptoethanol before testing permitted the differentiation of IgM and IgG antibodies, and possible differentiation of current infection from past exposure to the parasite. The antigen was stable over a 6-month evaluation period and also sh...
Williams R, Du Plessis DH, Van Wyngaardt W.Group-reactive enzyme-linked immunosorbent assays (ELISAs) were developed to selectively detect antibodies to African horsesickness virus (AHSV) and equine encephalosis virus (EEV), 2 orbiviruses that infect equids. In indirect ELISA, guinea pig antisera to all known AHSV or EEV serotypes recognized immobilized AHSV serotype 3 or EEV Cascara, respectively. Antisera from naturally infected animals did not cross-react with their respective heterologous viruses. The ELISA was used in parallel with the complement fixation (CF) and agar gel immunodiffusion tests to detect antibodies in sera from an...
Laviada MD, Arias M, Sánchez-Vizcaíno JM.The structural and non-structural proteins induced by African horsesickness virus serotype 4 (AHSV-4) in infected Vero cells were analysed by SDS-PAGE. Twenty-two virus-induced polypeptides were detected in infected cells by comparison with the polypeptides of mock-infected cells, of which four major (VP2, VP3, VP5 and VP7) and three minor (VP1, VP4 and VP6) structural proteins and four non-structural proteins (P58, P48, P21 and P20) were shown to be virus-coded, as deduced from electrophoretic and antigenic studies of purified virions and infected cells. The proteins that elicit the major ant...
Bürki F, Rossmanith W, Rossmanith E.Serological diagnosis of equine infectious anemia is of necessity group-reactive, i.e. based on viral core protein p26, because viral envelope components as well as the host's immune response to them undergo rapid antigenic change. Since 1970 the agar gel-immunodiffusion test ("Coggins-test") has been the diagnostic method of choice. Recently, ELISA tests have been introduced for faster and theoretically more sensitive serodiagnosis, while Western blots have been used to clarify doubtful results obtained in Coggins-tests. A commercial competitive ELISA was found to give practically equivalent ...
Gibson JS, O'Neill T, Thackray A, Hannant D, Field HJ.Serum antibody (virus neutralisation, complement fixation, IgM and IgG) responses to equine herpesvirus-1 (EHV-1) infection were measured in six foals which were initially free from EHV-1 and EHV-4 infection and maternally-derived antibodies. Following primary infection, high titres of virus neutralisation and complement fixation antibodies were detectable against EHV-1, however, corresponding antibody levels against EHV-4 were low or inapparent, although the two viruses share a number of cross-reactive epitopes. In addition, following the primary infection with EHV-1, IgM levels increased bef...
Adeyefa CA.Serological evidence of Equine Herpes virus type 1 (EHV-1) activity in Polo horses in Nigeria is reported for the first time. Eighty-two percent of horses tested with known antigen had precipitating antibodies to EHV-1 while 43% of sera tested against antigen prepared from nasal discharges were positive suggesting that the virus was being excreted in the nasal discharges and probably acting as a source of infection for incontact animals as occurs in on-going acute infections. The result of this study indicates a high prevalence of EHV-1 activity among Polo horses in Nigeria and demonstrates th...
Cohen ND, Carter GK, McMullan WC.Between Jan 1, 1984 and Aug 1, 1990, 27 horses were admitted to the veterinary medical center for evaluation of fistulous withers. Nine (37.5%) of 24 horses tested for antibody to Brucella abortus were seropositive. Horses that tested seropositive were significantly (P = 0.046) more likely to have been pastured with cattle that were seropositive for B abortus, and were significantly (P = 0.010) more likely to have had radiographic evidence of vertebral osteomyelitis than were horses that tested seronegative. Five horses that were seropositive for B abortus were administered strain 19 brucella ...
Chuma T, Le Blois H, Sánchez-Vizcaíno JM, Diaz-Laviada M, Roy P.The major core protein, VP7, of African horsesickness virus serotype 4 (AHSV-4), the aetiological agent of a recent outbreak of the disease in southern Europe, was expressed in insect cells infected with a recombinant baculovirus containing a cloned copy of the relevant AHSV gene (S7). Analyses of its biochemical and antigenic properties confirmed the authenticity of the protein expressed. The high-level expression of VP7 under the control of the strong polyhedrin promoter of Autographa californica nuclear polyhedrosis virus induced disc-shaped crystals in infected insect cells. This enabled u...
Adeyefa CA, Durojaiye OA.Serum samples obtained from 107 Polo horses showing clinical signs of viral respiratory disease were tested for precipitating antibodies to adenovirus by agar gel precipitation test and counter-immunoelectrophoresis method. The results obtained demonstrate serological evidence of adenovirus infection in Polo horses in Nigeria. The counter-immunoelectrophoresis method was observed to be about 3 times more sensitive than the agar gel precipitation test with 19.3 vs 64.5%. It could thus be used to screen a large number of serum samples within a short period.
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...
Weiland G, Hasslinger MA, Mezger S, Pöllein W.In an investigation period over 8 months the natural course of infection was studied by means of coproscopic and serological methods in 27 mares and 29 foals. The examination of the stool showed in mares, before the beginning of the grazing season, an infection rate of 100% with small and a rate of 7.4% with large strongyles (Str. vulgaris). Serologically the ELISA showed in foals only a distinct increase of antibody activity with the somatic antigen. The mares retained the high IgG-values of activity, which were already found at the beginning of the investigations. Even though the agglutinati...
Donahue JM, Smith BJ, Redmon KJ, Donahue JK.A study was conducted to evaluate a recently available fluorescent antibody test (FAT) conjugate for the detection of leptospires in tissues of aborted and stillborn horses, to determine the leptospira antibody titers and compare serologic test results with FAT results, and to determine the prevalence of leptospira-induced abortions and stillbirths in the equine population of central Kentucky. From July 1, 1988 through June 30, 1989, 15 (2.5%) of 594 submissions (fetuses, stillborn foals, and/or placentas) were diagnosed as leptospirosis by the FAT (14 of 15 tested) and/or microscopic agglutin...
Kölbl S, Schuller W, Pabst J.944 serum samples of horses, collected in 1988 and 1989, were examined for the occurrence of antibodies against equine arteritis virus by a microneutralizations test. In 10.9% of all sera reactors could be found. The distribution of seropositive horses varied from 4.6% (Salzburg) to 15.7% (Lower Austria). From Tyrol and Vorarlberg no samples could be obtained. It was not possible, to correlate clinical symptoms (infertility, respiratory symptoms, fever and edema) with the infection. It is assumed, that the disease appears in Austria only in a clinical inapparent form.
Rikihisa Y, Reed SM, Sams RA, Gordon JC, Pretzman CI.In August 1986, an extensive serosurvey for prevalence of IgG and IgM antibodies against Ehrlichia risticii, the causative agent of equine monocytic ehrlichiosis (EME), was performed at 2 Ohio racetracks, River Downs (RD) and Beulah Park (BP). Of 840 horses at RD and 574 at BP, 13 and 20%, respectively, were IgG antibody-positive (by indirect fluorescent antibody test results), with antibody titer ranging from 1:20 to 1:10,240. The titer observed at highest frequency at both racetracks was 1:80. A higher proportion of horses was ill at RD (operating during the summer months) than at BP (winter...
Verma RD, Sharma JK, Venkateswaran KS, Batra HV.A dot enzyme-linked immunosorbent assay (dot ELISA) was developed for diagnosis of glanders in equines. The test was based on the detection of IgG antibodies to Pseudomonas mallei antigens bound to nitrocellulose coated on plastic strips (dipsticks), the reaction being amplified by an avidin-biotin system with biotinylated anti-horse IgG and horseradish peroxidase-avidin D. Sera from 810 normal, six naturally infected and 48 sensitized equines were tested by this assay, and results were compared with complement fixation, indirect haemagglutination and counter-immunoelectrophoresis tests. Dot E...
Kaufman L, Mendoza L, Standard PG.Culture filtrate antigens of Basidiobolus ranarum and Conidiobolus coronatus were analyzed by immunodiffusion (ID) with homologous rabbit antisera. B. ranarum and C. coronatus were each found to have five specific antigens. Results of tests with heterologous antisera indicated that all of the species shared at least one antigen. ID tests incorporating the specific precipitin bands as references were developed for detection of basidiobolomycosis and conidiobolomycosis. These tests were performed with sera from humans and horses with proven basidiobolomycosis and conidiobolomycosis as well as wi...
Bürki F, Rossmanith E.Selected sets of serum samples of horses were tested blindly in a comparative investigation for antibodies against Equine Infectious Anemia (EIA) virus. Three commercial kits were used, a well-established agar-gel immuno-diffusion kit which our laboratory has been using routinely for 14 years on one hand, a competitive ELISA kit (CELISA) and a non-competitive ELISA kit on the other hand. The American EIA Reference Laboratory in Ames cotested 56 serum samples with the same 3 products, with highest-level correlation, thereby ascertaining full dependability of our own results. Five EIA experts su...
Madigan JE, Hietala S, Chalmers S, DeRock E.The prevalence of antibodies to Ehrlichia equi in horses from the foothill regions of northern California and from the Sacramento valley (non-foothill area) was determined, using an indirect fluorescent antibody test. Horses from foothill regions had a higher prevalence of seropositivity (10.4%) and higher titer (1:10 to 1:80) than did those from non-foothill regions (3.1%; titer less than or equal to 1:10). Fifty percent of healthy horses on a foothill farm enzootic for E equi had titer to E equi, suggesting that infection with E equi can be subclinical. Six veterinarians surveyed from northe...
Olchowy TW, Ames TR, Molitor TW.Antibody titer to Ehrlichia risticii was determined, in 2,549 equine serum samples, using an indirect fluorescent antibody assay. During 1986, samples were obtained from the Minnesota State-Federal Equine Infectious Anemia Diagnostic Laboratory, the Minnesota Racing Laboratory, from horses admitted to the University of Minnesota Veterinary Teaching Hospital, and as a result of field investigations of horses with acute diarrhea. Results of the study revealed antibody prevalence of 33, 24, 47, and 25% for the respective groups. There was no statistical association between seropositive status and...
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...
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...
Olchowy TW, Ames TR, Molitor TW.Antibody titer to Ehrlichia risticii was determined, in 2,549 equine serum samples, using an indirect fluorescent antibody assay. During 1986, samples were obtained from the Minnesota State-Federal Equine Infectious Anemia Diagnostic Laboratory, the Minnesota Racing Laboratory, from horses admitted to the University of Minnesota Veterinary Teaching Hospital, and as a result of field investigations of horses with acute diarrhea. Results of the study revealed antibody prevalence of 33, 24, 47, and 25% for the respective groups. There was no statistical association between seropositive status and...
Zhou EM, Riva J, Clavijo A.An immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (MC-ELISA) was developed for the detection of primary infection of vesicular stomatitis virus (VSV) in equine and swine sera. The test was based on the use of biotinylated sheep antibodies against equine or swine IgM molecules bound to a streptavidin-coated ELISA plate. The captured IgM antibodies were detected by application of antigens prepared from the New Jersey and the Indiana VSV serotypes (VSV-NJ and VSV-IN, respectively) and mouse polyclonal antibodies against VSV-NJ and VSV-IN. The MC-ELISA was compared to a competiti...
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...
Fikrig E, Magnarelli LA, Chen M, Anderson JF, Flavell RA.Enzyme-linked immunosorbent assays (ELISA) and immunoblots using either whole-cell lysates of Borrelia burgdorferi or an antigenic region of flagellin (41-G) as the antigen were performed, and the abilities of the two assays to detect antibodies to this spirochete in dog, cottontail rabbit, and horse sera were compared. Assays using whole-cell B. burgdorferi lysates as the antigen were more sensitive for detecting antibodies. ELISA with 41-G as the antigen were specific for Borrelia antibodies but were not as sensitive as the assays with whole-cell lysates coated to the solid phase. Use of rec...
Traversa D, Milillo P, Maggi R, Simonato G, Di Cesare A, Pezzuto C, Grillini M, Morelli S, Colombo M, Passarelli A, Grassano A, Serio P, Losurdo M....Equine and canine anaplasmosis and borreliosis are major tick-borne zoonotic diseases caused by and various species of (the most important being s.l.), respectively. This study evaluated the seroexposure to and in dogs and horses used in Animal-Assisted Interventions or living in contact with children, elderly people or immunocompromised persons. A total of 150 horses and 150 dogs living in Italy were equally divided into clinically healthy animals and animals with at least one clinical sign compatible with borreliosis and/or anaplasmosis (present at clinical examination or reported in th...
Verney M, Gautron M, Lemans C, Rincé A, Hans A, Hébert L.Trypanozoon infections in equids are caused by three parasite species in the Trypanozoon subgenus: Trypanosoma equiperdum, T. brucei and T. evansi. They are respectively responsible for infectious diseases dourine, nagana and surra. Due to the threat that Trypanozoon infection represents for international horse trading, accurate diagnostic tests are crucial. Current tests suffer from poor sensitivity and specificity, due in the first case to the transient presence of parasites in the blood and in the second, to antigenic cross-reactivity among Trypanozoon subspecies. This study was designed to...
Daddy KK, Mwanza M, Oguttu JW, Ngoma L.Leptospirosis is one of the major emerging global economic and health problems affecting donkeys, thereby reducing their work output. Furthermore, the disease has public health importance because of its zoonotic nature. Despite the significant contribution donkeys make to the national economy, less attention is given to diseases that afflict donkeys and reduce their productivity and performance. A cross-sectional study was conducted to investigate the seroprevalence of Leptospira and identify risk factors associated with the occurrence of the disease among donkeys in the study area. Methods: A...
Hobo S, Niwa H, Anzai T.The reactivity of the proline-glutamic acid-proline-lysine (PEPK) repetition peptide antigen in 3176 serum samples was investigated to evaluate its utility as an antigen for the serological diagnosis of strangles. The reactivity of the sera of horses infected with Streptococcus equi subspecies equi was high when the peptide had several PEPK repetitions. However, as the number of PEPK repetitions increased, the reactivity of the antigen with the sera of horses infected with Streptococcus equi subspecies zooepidemicus also increased. In horses infected experimentally with S equi, the reactivity ...
Bürki F, Rossmanith E.Selected sets of serum samples of horses were tested blindly in a comparative investigation for antibodies against Equine Infectious Anemia (EIA) virus. Three commercial kits were used, a well-established agar-gel immuno-diffusion kit which our laboratory has been using routinely for 14 years on one hand, a competitive ELISA kit (CELISA) and a non-competitive ELISA kit on the other hand. The American EIA Reference Laboratory in Ames cotested 56 serum samples with the same 3 products, with highest-level correlation, thereby ascertaining full dependability of our own results. Five EIA experts su...
Huang X, Xuan X, Yokoyama N, Katayama Y, Anzai T, Igarashi I.Two enzyme-linked immunosorbent assays (ELISA) with recombinant protein as antigens were evaluated by comparison with the indirect fluorescent antibody tests (IFAT) for the detection of specific antibodies to Babesia caballi and Babesia equi, respectively in 380 sera from experimentally infected, uninfected, and field horses. The high concordances of 92.4% (351/380) and 98.2% (373/380) between ELISA and IFAT for B. caballi and B. equi, respectively suggest that ELISA, especially for B. equi infection, could be alternative to the corresponding IFAT for serodiagnoses of equine piroplasmosis, alt...
Alshammari A, Gattan HS, Marzok M, Salem M, Al-Jabr OA, Selim A.Fasciola hepatica is a trematode parasite that affects wide range of mammalian hosts including horses. There are no epidemiological data on fasciolosis in Egyptian horses. Hence, the present study aimed to evaluate the seroprevalence of F. hepatica in horses using ELISA as well as assess the associated risk factors for fasciolosis infection. A total of 400 sera samples of horses from three Egyptian governorates were examined. The overall seroprevalence rate in horses was 13% and the disease was more prevalent in Qalubia (14.17%) and Gharbia (16.67%) which are located at Nile Delta. The likelih...
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...
Kweon CH, Kwon BJ, Ko YJ, Kenichi S.VP7, the sero-group common antigen, of African horsesickness virus (AHSV-4) was expressed in insect cells by recombinant baculovirus. To develop a specific diagnostic method, monoclonal antibody (Mab) against VP7 was prepared and investigated as diagnostic reagent with the baculovirus expressed VP7. However, the Mab against VP7 of AHSV cross-reacted with Chuzan virus by the indirect immunofluorescence assay (IFA), confirming the presence of conserved domain of VP7 among Orbiviruses. This study describes two types of ELISA; Mab linked indirect (I-ELISA) and competitive-ELISA (C-ELISA) using bac...
Singh BK, Ahuja S, Gulati BR.A single-dilution, sensitive and specific monoclonal antibody-based blocking enzyme-linked immunosorbent assay (B-ELISA) was developed as an alternative to the cumbersome virus neutralization test (VNT) for detection of equine herpesvirus-1 (EHV-1) antibodies. Neutralizing monoclonal antibodies (1H6 and 9C6) raised against EHV-1 (Hisar-90-7 strain) and sera from 70 horses (30 known negative and 40 known positive for EHV-1 antibodies by VNT) were used for standardization of the B-ELISA. Using a single serum dilution of 1:250 in B-ELISA, 100% specificity was obtained with both monoclonal antibod...
Wang Y, Zhang LP, Li J, Li DD, Zhang Q, Li C.The aim of this study was to determine the seroprevalence of Babesia caballi and Theileria equi in horses imported into Shanghai port. Between 2018 and 2019, 344 horse sera samples were collected and tested for B. caballi and T. equi, using commercially available kits. Only one B. caballi seropositive sample was detected. To the best of our knowledge, this is the first report of a B. caballi seropositive in imported horses at Shanghai port, which reflects the importance of monitoring piroplasmosis seroprevalence in imported horses.
Karamalla ST, Gubran AI, Adam IA, Abdalla TM, Sinada RO, Haroun EM, Aradaib IE.African horse sickness virus (AHSV) is an infectious non contagious insect-transmitted double-stranded (ds) RNA orbivirus of the family Reoviridae. AHSV causes an often fatal hemorrhagic infection with high mortality among selected breeds of Arabian horses. This study was conducted to avail some information with regard to the prevalence and associated risk factors of AHSV among ecotype breeds of horses in central Sudan. Methods: Sera were collected from 320 horses, which were selected randomly from four localities and employed in the study. A competitive enzyme-linked immunosorbent assay (cELI...
Pinna A, Martins G, Souza G, Lilenbaum W.The aim of this study was to demonstrate that seroreactivity against Leptospira is significantly associated to the reproductive efficiency of recipient mares of an embryo transfer (ET) programme. A serosurvey was conducted from August 2007 to March 2009 in five herds from Rio de Janeiro, Brazil, with high rates of reproductive failure, as early embryonic death (>12%), abortion (>12%) and perinatal death. Detailed information about the losses was obtained from practitioner. A total of 338 recipient mares were tested by microscopic agglutination test, and 226 (66.9%) were seroreactive, mainly ag...
Sajid M, Ahmad MU, Khan MA, Anjum MA, Mushtaq MH.The present study was an attempt to elucidate the seroprevalence of equine influenza virus (H3N8) in two geographically distinct regions of Pakistan where vaccination is not routinely practiced. Methods: A total number of 315 animals of family Equidae were included in the survey. Blood samples and nasal swabs were collected from the same animal and analyzed through ELISA and Hemagglutination Inhibition. Results: The seroprevalence for EIV was 10 and 8.39 in districts under study. Out of 29 ELISA positive sera, 7 (24.1 %) showed antibodies against H1 and 22 (75.9 %) showed against H3 of influen...
Joncas JH, Gilker JC, Chagnon A.The relative value of heterophil agglutinins (HA) and of specific EBV antibodies in the diagnosis of infectious mononucleosis (IM) was assessed in 108 cases of the disease and in 280 controls. Among the 108 cases 93 were HA-positive by sheep cells in at least one of their sera, while 15 were HA-negative by the same test. Among the 280 controls false-positive HA tests were not encountered except in eight cases with the horse cell microtitre tests. With one of the two slide tests at least two false-positive tests and 12 false-negative tests were also found but these sera had low titres in microt...
Phumoonna T, Muscatello G, Chicken C, Gilkerson JR, Browning GF, Barton MD, Heuzenroeder MW.A total of 227 field samples from naturally exposed foals aged between 3 weeks and 6 months were used in an evaluation of a peptide-based enzyme-linked immunosorbent assay (ELISA) for diagnosis of Rhodococcus equi infection. A biotinylated peptide derived from the virulence-associated protein A (VapA) of R. equi, a horse pathogen, was synthesized and designated as PN11-14. The peptide corresponds to the N-terminal B-cell epitope TSLNLQKDEPNGRASDTAGQ of the VapA protein. Based upon a serum immunoglobulin (Ig)G titre of 512 as a positive cut-off value for the R. equi infection, the ELISA provide...
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 ...
Crabb BS, Studdert MJ.A series of truncated equine herpesvirus 1 (EHV1) glycoprotein C (gC) molecules was examined for use as serodiagnostic antigens for EHV1 and EHV4. Small regions of EHV1 glycoprotein C, an immunodominant EHV1 glycoprotein, were expressed in Escherichia coli as glutathione S-transferase (GST) fusion proteins using the bacterial expression vector pGEX-2T. Sera obtained from horses, including sera from specific-pathogen-free (SPF) foals, following exposure to either EHV1, EHV4 or both viruses were used. Several of the fusion proteins were shown to encompass EHV1 specific epitopes while others enco...
Wernery U, Chan E, Raghavan R, Teng JLL, Syriac G, Siu SY, Joseph M, Yeung ML, Jia L, Cai JP, Chiu TH, Lau SKP, Woo PCY.Glanders is a highly contagious and potentially serious disease caused by Burkholderia mallei, a Tier 1 select agent. In this study, we raised a monoclonal antibody (mAb) against the lipopolysaccharide (LPS) of B. mallei and developed a competitive enzyme-linked immunosorbent assay (cELISA) for B. mallei infection. Using the titrated optimal conditions of B. mallei-LPS (2 ng) for microtiter plate coating, sample serum dilution at 1:20 and 3.5 ng/μL anti-LPS mAb B5, the cutoff value of the cELISA was determined using serum samples from 136 glanders-free seronegative horses in Hong Kong. All ca...
Schvartz G, Epp T, Burgess HJ, Chilton NB, Pearl DL, Lohmann KL.Equine granulocytic anaplasmosis (EGA) and Lyme borreliosis (LB) are an emerging concern in Canada. We estimated the seroprevalence of EGA and equine LB by testing 376 convenience serum samples from 3 provinces using a point-of-care SNAP(®) 4Dx(®) ELISA (IDEXX Laboratories, Westbrook, Maine, USA), and investigated the agreement between the point-of-care ELISA and laboratory-based serologic tests. The estimated seroprevalence for EGA was 0.53% overall (0.49% in Saskatchewan, 0.71% in Manitoba), while the estimated seroprevalence for LB was 1.6% overall (0.49% in Saskatchewan, 2.86% in Manitob...
Jahrling PB, Hesse RA, Metzger JF.A radioimmunoassay (RIA) procedure is described for measuring antibodies to alphaviruses in human and other mammalian sera. The test employed protein Abearing Staphylococcus aureus as a solid-phase immunoadsorbent for (3)H-labeled viruses complexed with immunoglobulin G. Using antibodies produced in humans and guinea pigs, the RIA procedure clearly differentiated among antibodies to Venezuelan, western, and eastern equine encephalomyelitis viruses. Sensitivity of the RIA depended on the concentrations of labeled viruses employed. The dilution of serum that effected binding of 50% of the (3)H-l...
Sánchez-Matamoros A, Nieto-Pelegrín E, Beck C, Rivera-Arroyo B, Lecollinet S, Sailleau C, Zientara S, Sánchez-Vizcaíno JM.African horse sickness (AHS) is considered a fatal re-emergent vector-borne disease of horses. In the absence of any effective treatment for AHS, vaccination remains the most effective form of disease control. The new generation of vaccines, such as one based on purified, inactivated AHS virus (AHSV, serotype 4), which does not induce antibodies against non-structural protein 3 (NS3), enables the development of diagnostic methods that differentiate infected from vaccinated animals (DIVA assays). As detecting AHS in AHSV-free countries may lead to restrictions on international animal movements ...
Sahu A, Dhanze H, Singh V, Mehta D, Gupta M, Singh M, Vinod VK, Gulati BR.Japanese encephalitis (JE) is a re-emerging mosquito borne disease, for which equines are most susceptible amongst all animals. Detection of specific immunoglobulin 'M' (IgM) is considered as an ideal way to diagnose recent JE virus infection in equines due to low virus load and short-term viremia. The present study was undertaken to develop a sensitive and specific recombinant NS1 protein based indirect IgM-ELISA and IgM capture (MAC) ELISA to diagnose recent infection of JEV in equines. Indirect IgM ELISA was standardized with relative diagnostic sensitivity and specificity of 100% and 88.5%...
Singha H, Goyal SK, Malik P, Singh RK.Thermostabilizing effect of heavy water (D2O) or deuterium oxide has been demonstrated previously on several enzymes and vaccines like oral poliovirus vaccine and influenza virus vaccine. In view of the above observations, effect of heavy water on in situ thermostabilization of recombinant p26 protein on enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of equine infectious anemia virus (EIAV) infection was investigated in the present study. The carbonate-bicarbonate coating buffer was prepared in 60% and 80% D2O for coating the p26 protein in 96-well ELISA plate and thermal stabilit...
Meny P, Iglesias T, Menéndez C, Quintero J, Ríos C, Ashfield N, Ferreira O, Mosca V, De Brun L, Ortiz G, De Vries I, Varela G, Schelotto F.To investigate seroprevalence of anti-Leptospira antibodies in equines and associated workers in Uruguay, 891 equine and 150 human sera were drawn; 212 equine urine samples were also taken for culture. Environmental conditions and equine raising or managing practices were recorded in all 72 visited establishments; epidemiological information was obtained from each worker. Microscopic agglutination technique (MAT) was performed with 10 Leptospira strains for equines and 18 for human sera, that were also studied with IgM indirect immunofluorescence (IgM-IIF). Equine titres ≥100 were considered...
Obregón AM, Fernández C, Rodríguez I, Balbis Y, Martínez B, Rodríguez J.To assess the sensitivity, specificity, reproducibility, and stability of five latex agglutination systems for detecting antibodies against leptospira in human and animal sera, by using the Leptospira serotypes that are most widely prevalent in Cuba. Methods: We performed an analytic and descriptive study with 706 human sera (65 tested positive for antibodies against leptospira with microagglutination (MAT) and hemagglutination (HA) techniques; 156 sera that tested negative with MAT and HA); 485 sera from 424 patients who had clinical or epidemiologic signs of leptospirosis; and 29 animal sera...
