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Topic:Serodiagnosis
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.
Prevalence of equine piroplasmosis in Central Mongolia. Antigen for the indirect fluorescent antibody test (IFAT) was routinely prepared from infected erythrocytes from horses experimentally infected with Babesia equi and Babesia caballi. With the successful establishment of in vitro cultures of B. equi and B. caballi, it is now possible to employ culture-derived antigens in this test. In this study, in vitro-propagated B. equi- and B. caballi-infected erythrocytes were used as antigen in the IFAT. Various modifications to an established protocol had to be implemented to allow repeatable results. Cultures with 3-4% parasitized erythrocytes were fou...
Serodiagnosis of Babesia equi infection–a comparison of Dot-ELISA, complement fixation test and capillary tube agglutination test. 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...
Seroprevalence of antibodies to Sarcocystis neurona in horses residing in a county of southeastern Pennsylvania. To determine seroprevalence of Sarcocystis neurona-specific antibodies in a population of horses residing in Chester County, Pa. Methods: Prevalence survey. Methods: 117 serum samples from selected members of a population of 580 Thoroughbred horses. Methods: Serum was analyzed for antibodies to Sarcocystic neurona, using a western blot. Information regarding age, sex, and housing of horse was obtained by questionnaire. Data were analyzed, using multivariable logistic regression. Results: Seroprevalence was 45.3% (95% CI, 36.3 to 54.3%). A relationship was not found between seroprevalence and s...
Serologic markers in early stages of African horse sickness virus infection. Fifteen horses were experimentally infected with African horse sickness virus (AHSV) serotype 4. To learn more about the time course of production and specificity of AHSV-specific antibodies, sera were analyzed by immunoblot analysis. Only animals that survived for more than 9 days were able to develop a humoral immune response detectable by immunoblotting. The earliest serological markers corresponded mainly to VP5, VP6, and NS2 and to a lesser extent to VP3, NS1, and NS3. Neutralizing antibodies to VP2 were not detected by immunoblotting, suggesting that they are mostly conformation dependen...
Serologic diagnosis of canine and equine borreliosis: use of recombinant antigens in enzyme-linked immunosorbent assays. Serum samples from dogs and equids suspected of having canine or equine borreliosis, respectively, were analyzed in polyvalent enzyme-linked immunosorbent assays (ELISAs) with whole-cell or recombinant antigens of Borrelia burgdorferi sensu stricto. Purified preparations of recombinant antigens included outer surface protein A (OspA), OspB, OspC, OspE, OspF, and p41-G (a fragment of flagellin). Of the 36 dog sera that reacted positively to whole-cell antigen, 32 (88.9%) contained antibodies to one or more recombinant antigens. Reactivities to OspF (88.9% positive) and p41-G (75% positive) were...
Application of equine infectious anemia virus core proteins produced in a baculovirus expression system to serological diagnosis. 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...
A sero-epidemiological survey of equine piroplasmosis in the northern and eastern Cape Provinces of South Africa. Serum samples from yearling Thoroughbred horses (n = 176) in the magisterial districts of Colesberg, Venterstad, and Wodehouse in the Northern and Eastern Cape Provinces were collected between September and November 1995 to determine the prevalence of antibodies to Babesia equi and Babesia caballi in these regions. Samples were examined for specific antibodies using the indirect fluorescent antibody test. The 95% confidence intervals for the prevalence of serum antibodies in the 3 districts combined varied from 47% to 61% for B. equi and from 26% to 40% for B. caballi. Antibody prevalence did ...
Epidemiological aspects of the Brazilian spotted fever: serological survey of dogs and horses in an endemic area in the State of São Paulo, Brazil. In order to obtain information on Brazilian spotted fever, a study in domestic animals was performed in the County of Pedreira, State of São Paulo, Brazil, where 17 human cases had been notified. Serum samples obtained from animals were tested by indirect immunofluorescence for detectable antibodies to spotted fever-group rickettsiae. Seropositivity was revealed in 12 (36.4%) of 33 dogs and seven (77.8%) of nine horses from the endemic area. For comparison, blood samples from dogs and horses from non endemic area were tested and four (12.9%) of 31 dogs and three (27.3%) of 11 horses were posi...
Equine herpesvirus type 2: prevalence and seroepidemiology in foals. Whole blood and serum were collected from foals to determine the prevalence of Equine herpesvirus type 2 (EHV 2) infection in foals, age at which infection can first be identified and serological responses to infection. Equine herpesvirus type 2 was isolated from peripheral blood mononuclear cells (PBMC) from 68 of 69 foals, 1-8-months-old, sampled once. Virus isolation was performed twice at intervals of 2-7 months on PBMCs from 33 foals and EHV2 was isolated on both occasions in all but one foal (negative, then positive). Regression analysis of log2-transformed reciprocal serum EHV2 virus ne...
Serologic responses to Rhodococcus equi in individuals with and without human immunodeficiency virus infection. Thirty healthy blood donors, 15 workers from horse-breeding farms, 69 human immunodeficiency virus (HIV)-negative persons at risk for HIV infection, 125 HIV-infected subjects without Rhodococcus equi infection, and nine HIV-infected patients with Rhodococcus equi pneumonia were evaluated in order to detect serum antibodies to Rhodococcus equi precipitate-soluble antigen by an enzyme immunoassay (EIA). Whereas EIA values for healthy donors, horse farm workers, individuals at risk for HIV infection, and HIV-positive subjects without Rhodococcus equi infection were comparable, HIV-infected patien...
Evaluation of travel and use as a risk factor for seropositivity to Ehrlichia risticii in horses of New York state. To determine whether mean annual frequency and destination of equine travel was associated with exposure to Ehrlichia risticii and whether these associations were modified by horses' place of residence. Methods: Cross-sectional study. Methods: 511 equine operations containing 2,587 horses were visited in New York state from a target population of 39,000 operations. Methods: Each horse was tested for serum antibodies against E risticii, using indirect fluorescent antibody. Information on the horse's travel history, farm's management practices, and surrounding ecology was obtained by personal in...
Cross-sectional evaluation of environmental, host, and management factors associated with risk of seropositivity to Ehrlichia risticii in horses of New York state. To locate counties within New York state with a high seroprevalence among the equine population, to determine host, management, and environmental factors that were associated with seropositivity to Ehrlichia risticii, and to determine evidence for arthropod- or helminth-mediated transmission of E risticii to horses. Methods: Cross-sectional study. Methods: A random sample of 3,000 of the 39,000 equine operations in New York state was selected, and 2,587 horses from 511 operations were tested. Methods: Blood samples were collected from horses and tested for seropositivity, using the indirect fl...
Equine piroplasmosis an update on diagnosis, treatment and prevention. Two haemoprotozoan parasites, Babesia caballi and Babesia equi, can cause equine piroplasmosis. Due to the presence of potential tick vectors in areas so far unaffected by equine babesias, import and export regulations often require the serum testing of animals for evidence of infection. Although the complement fixation test (CFT) has been recommended for detecting the presence of antibodies to Babesia spp., it has been demonstrated to have several disadvantages, including false-positive results and low sensitivity for detecting latent infections. An enzyme-linked immunosorbent assay (ELISA) m...
Use of excretory/secretory antigens for the serodiagnosis of Anoplocephala perfoliata cestodosis. Whole worm extract (WWE) and excretory/secretory (E/S) antigens of Anoplocephala perfoliata were characterised by SDS-PAGE and their use in the serodiagnosis of equine cestodosis was evaluated. An enzyme-linked immunosorbent assay (ELISA) was used to compare WWE and E/S antigen as the capture layer in an antibody capture ELISA. E/S antigen gave the best differentiation between sera from tapeworm-positive and tapeworm-negative horses. The E/S-ELISA was optimised and validated against sera from horses of known tapeworm status. This assay gave a diagnostic sensitivity of 68% (n = 38) and a specif...
Evidence for a high rate of false-positive results with the indirect fluorescent antibody test for Ehrlichia risticii antibody in horses. The original objective was to determine seroprevalence of Ehrlichia risticii antibody among horses in California. On the basis of the unexpected results of the survey, an investigation into the accuracy and reproducibility of results of the indirect fluorescent antibody (IFA) test for E risticii was carried out. Methods: Prospective, seroprevalence study. Methods: Healthy horses (n = 655) and horses with clinical signs of equine monocytic ehrlichiosis (EME; n = 514) from various regions of California. Methods: The IFA test was performed. Results were compared with results of an ELISA and with ...
Association of leptospiral seroreactivity and breed with uveitis and blindness in horses: 372 cases (1986-1993). Recurrent uveitis, a leading cause of blindness in horses, often develops as a sequela to systemic leptospirosis. Over a 7-year period, 63 of 112 (56%) horses with uveitis were seropositive for Leptospira interrogans serovar pomona, but only 23 of 260 (9%) horses without uveitis were seropositive. Odds-ratio analysis revealed that seropositive horses were 13.2 times more likely to have uveitis than were seronegative horses. Of the 63 seropositive horses with uveitis, 59% developed blindness, compared with only 24% in the 49 seronegative horses with uveitis that lost vision in 1 or both eyes du...
Equine viral arteritis with abortions: serological and virological evidence in Germany. An outbreak of clinical disease with abortions, suspected to be equine viral arteritis in Germany was examined serologically and virologically. Three of five animals tested seroconverted to EAV, and EAV was isolated from an aborted fetus. In view of the significant increase in seropositive horses, the need for control measures is emphasized.
Expression of small regions of equine herpesvirus 1 glycoprotein C in Escherichia coli. 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...
Seroanalysis of Tyzzer’s disease in horses: implications that multiple strains can infect Equidae. A monoclonal antibody based competitive inhibition assay was used to detect antibodies in horse sera to purified flagellar antigens from distinct Clostridium piliforme isolates. Sequential absorption of hyperimmune rat serum to C. piliforme isolate E (horse-origin isolate), a positive C. piliforme-immune horse serum, and other suspected immune horse sera with unrelated bacteria or C. piliforme isolates E or isolate R1 (rat-origin isolate) alone demonstrated the specificity of this assay for C. piliforme. This specificity was associated with the inhibition of monoclonal antibody binding to C. p...
Prevalence and serovars of leptospira involved in equine abortions in central Kentucky during the 1991-1993 foaling seasons. In this study, the prevalence of leptospira-induced abortions/stillbirths for the past 3 foaling seasons (1991-1993) was determined, and fetal tissues and/or the mare's urine from positive cases were cultured in an attempt to isolate and identify the leptospira serovars responsible for the abortions. The sensitivity and specificity of the primary diagnostic tests, the fluorescent antibody test (FAT) and the microscopic agglutination test (MAT), used for the diagnosis of leptospirosis were also determined. For the 3 years, 74 (3.3%) of 2,264 abortion/stillborn submissions were diagnosed as lept...
Investigations on naturally occurring Trypanosoma evansi infections in horses, cattle, dogs and capybaras (Hydrochaeris hydrochaeris) in Pantanal de Poconé (Mato Grosso, Brazil). The prevalence of Mal de Cadeiras--Portuguese for Trypanosoma (T.) evansi infections in horses--as well as the prevalence of T.evansi infections in cattle, dogs and free-ranging capybaras (Hydrochaeris hydrochaeris) was investigated in Pantanal de Poconé (Mato Grosso, Brazil). In 0.3, 8.6 and 8.0% of the horses, dogs and capybaras, respectively, infection was detected using standard parasitological methods. A seroprevalence of 4.1, 2.3, 7.1 and 22.0% was found in horses, cattle, dogs and capybaras, respectively, using an enzyme-linked immunosorbent assay for the detection of T.evansi antigen ...
Monoclonal antibody based competitive ELISA for the detection of specific antibodies against Coxiella burnetii in sera from different animal species. A competitive ELISA system for the detection of antibodies against Coxiella (C.) burnetii in cattle, sheep, goats, horses and humans is described. The ELISA is based on a biotinylated monoclonal antibody with specificity for C. burnetii lipopolysaccharide in combination with streptavidin peroxidase. For evaluation and statistical analysis, 413 sera from cattle, sheep, goats, horses and humans were tested in parallel in the indirect immunofluorescence test (IFT). Furthermore, a total of 448 bovine and human sera were also tested with an indirect ELISA and 47 sheep sera were investigated using t...
Borrelia burgdorferi infection in UK horses. Antibody levels (IgG and IgM) to Borrelia burgdorferi were measured in the sera and synovial fluids of UK horses. Western blotting against B. burgdorferi was also used on samples from seropositive horses. A low incidence of seropositivity was shown in horses from most parts of the UK. This increased in areas that have a high incidence of human and canine borreliosis (Norfolk and south coast). Leptospira infections of horses did not cause cross reactions in the B. burgdorferi ELISA. Most horses did not display clinical signs of Lyme disease. As with dogs and man, it is apparent that B. burgdorf...
Diagnosis of Babesia caballi infections in horses by enzyme-linked immunosorbent assay (ELISA) and western blot. From Babesia caballi in vitro cultures a preparation of 100% infected erythrocytes was obtained. From this, B. caballi antigens were extracted with the detergent 3-[(3-Cholamidopropyl)-dimethylammonio]-1-propane-sulfonate (CHAPS) and used as ELISA antigens. A control antigen of normal erythrocytes from the same donor horse was prepared in an identical manner. The ELISA and Western blot were validated by testing of sera from horses experimentally infected with B. caballi or B. equi or not infected with Babesia spp. ELISA and Western blot results were compared with those obtained by the immunofl...
Identification of diagnostic antigens for South American Babesia caballi infections. 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...
Geographical variation of seropositivity to Ehrlichia risticii (equine monocytic ehrlichiosis) of horses in New York state. A total of 2,579 serum samples from horses in New York state during 1985-1986 were examined for seropositivity to Ehrlichia risticii using the indirect fluorescent antibody technique. Cluster analysis statistical technique was used to group counties according to their estimated EME-disease rate (seropositive proportion of sampled horses). Counties were clustered into 4 groups of different EME-disease rates, representing high (86% seropositive), medium (66% seropositive), medium-low (47% seropositive) and low (6% seropositive) risk regions. The logistic regression statistical technique was used...
Serological and microbiological findings on 3 farms with equine leptospiral abortions. Blood and urine samples from horses on 3 central Kentucky horse farms with prior histories of leptospiral abortions were analysed. Blood samples were obtained from all available horses on each farm and tested for antibodies to 6 leptospira serovars. Urine samples were collected from non-gravid mares with serum antibody titres > or = 1:800 and examined for leptospires by dark-field microscopy, fluorescent antibody testing and culture. Adult horses had the greatest serological evidence of exposure to leptospira, followed by yearlings, then foals. Of horses with anti-leptospiral antibodies, 76...
[Rhinopneumonia and equine viral arteritis: seroepidemiological study in the northeast of Tunisia]. 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.
