Immunofluorescence assay (IFA) is a laboratory technique used to detect and visualize specific antigens or antibodies in equine tissue samples or bodily fluids. This method employs fluorescent-labeled antibodies to bind target molecules, allowing for the observation of fluorescence under a microscope. In horses, IFA is employed in various research and diagnostic applications, including the study of infectious diseases, immune responses, and cellular localization of proteins. The technique provides valuable insights into the distribution and expression of specific proteins within equine cells and tissues. This page aggregates peer-reviewed research studies and scholarly articles that explore the methodology, applications, and advancements of immunofluorescence assay in equine research.
Hodgin EC, Miller DA, Lozano F.Leptospira infection was diagnosed as the cause of 4 late-term equine abortions/stillbirths and 1 neonatal death in Louisiana. The most consistent gross and microscopic lesions were icterus and interstitial nephritis, respectively. Diagnoses were based on visualization of compatible spirochetes in Warthin-Starry-stained sections of kidney, liver, and placenta. Confirmation by immunofluorescence was made in 2 cases.
Budras KD, Hullinger RL, Sack WO.The laminar epidermis (epidermis parietis) of hooves from 14 clinically normal horses, 6 months to 15 years old, was examined by light and electron microscopy and immunofluorescence to measure the contributions of this region to the formation of the hoof wall. By their progressive keratinization to form primary epidermal laminae, the secondary epidermal laminae ultimately contributed about 20% of the thickness of the hoof wall (as revealed in the white line [zona alba]). The keratinized, primary epidermal laminae were developed to a height of 4 mm during their proximodistal-course, much of thi...
Soule C, Dupouy-Camet J, Georges P, Ancelle T, Gillet JP, Vaissaire J, Delvigne A, Plateau E.Three groups of three horses each were, respectively, infected with 5000, 20,000 and 50,000 larvae of Trichinella spiralis. The strain used was isolated from a human biopsy during horsemeat-related outbreaks of trichinellosis in France. Transient muscular disorders were only observed in two of the horses infected with 50,000 larvae but none of the horses had fever. A significant increase in blood eosinophils was noticed in 5 horses. Serum LDH, aldolase and CPK peaked at the fifth week post-infection. Specific IgG assayed by indirect immunofluorescence and ELISA, appeared 2-5 weeks post-infecti...
Brostroöm H, Paulie S, Perlmann P.To characterise the expression of surface antigens on equine sarcoid cells compared to normal equine fibroblasts, immune sera were produced in rabbits against transformed cells of a virus-containing sarcoid cell line (Mc-1) and normal dermal fibroblasts, respectively. The specificities of the sera were analysed by antibody-dependent cellular cytotoxicity against 51Cr-labelled target cells using human lymphocytes as effector cells. Anti-Mc-1 antiserum induced strong cytotoxicity against transformed cells of two sarcoid cell lines (Mc-1 and Bay Mc-1), whereas the cytotoxicity against transformed...
ter Laak EA, Fennema G, Jaartsveld FH.Contagious Equine Metritis (CEM) was detected in the Netherlands for the first time in 1987. A total number of five mares (Dutch saddle-horse) were infected in three separate outbreaks. The origin of the infection could not be determined in any of the cases. As the isolates of the causal organism, Taylorella equigenitalis, showed auto-agglutination, diagnosis was difficult. Therefore, an indirect immune fluorescence test as used to diagnose the second isolate. Five strains were isolated, which all were resistant to streptomycin. The prevalence of CEM since 1981 is summarised. The importance of...
Blach EL, Amann RP, Bowen RA, Frantz D.Better procedures for freezing and thawing equine sperm are needed since variable fertility is obtained when cryopreserved sperm are used. To evaluate current methods of freezing equine sperm, we examined spermatozoal quality by means of two new techniques. These measured the integrity of plasma-acrosomal membranes by immunofluorescent analyses of binding of an antibody specific to the acrosome and evaluated eight parameters of spermatozoal motion using a fully automated computerized system. Five ejaculates from each of eight stallions were processed for freezing in egg yolk-lactose extender w...
Liu IK, Bernoco M, Feldman M.Ten fertile feral mares and 6 domestic horses (4 fertile mares, 1 infertile mare, 1 gelding) were immunized with heat-solubilized pig zonae pellucidae by 4 injections equivalent to 2000 or 5000 zonae each at 2-4-week intervals and a booster injection of 20,000 zonae 6-10 months after the last of the initial inoculations. The immune response was reflected by high antibody levels as measured by an enzyme-linked immunosorbent assay (ELISA) using immobilized pig zona antigen. In-vivo inhibition of fertility occurred in 12 (86%) of the 14 fertile mares studied and persisted for a minimum of 7 month...
Cecio A, Vittoria A.Amines and/or peptide-producing cells, deserving to be called paraneurons, were demonstrated in the urethro-prostatic complex of the man, rabbit, buffalo and sheep and in the uterine horns of the pig, horse and mouse, by means of histochemical, immunohistochemical, immunofluorescent and double labeling immunofluorescent techniques. In particular, the urethro-prostatic complex of the sheep contains cells producing serotonin, chromogranin A and somatostatin. Often the amine and the "marker"-protein were colocalized in the same cells. Chromogranin A- and somatostatin-containing cells were found i...
Karabatsos N, Lewis AL, Calisher CH, Hunt AR, Roehrig JT.A virus, strain 64A-1519, isolated from the brain of a horse dying of encephalitis in Florida in 1964, was identified as western equine encephalomyelitis (WEE) virus. Recently, we used polyclonal and monoclonal immune reagents to identify this isolate by comparing it to 2 strains of WEE virus and to Highlands J (HJ) virus in hemagglutination-inhibition, immunofluorescent antibody, and plaque-reduction neutralization tests. These tests demonstrate that strain 64A-1519 is a strain of HJ virus distinct from WEE virus.
Rikihisa Y, Pretzman CI, Johnson GC, Reed SM, Yamamoto S, Andrews F.Ehrlichia risticii has a close antigenic relationship to E. sennetsu. Sera of ponies experimentally infected with E. risticii, the etiologic agent of Potomac horse fever, consistently reacted with E. sennetsu, a human pathogen, in indirect fluorescent-antibody (IFA) testing, while human E. sennetsu convalescent serum reacted with E. risticii by IFA testing and immunoferritin labeling of cells infected in vitro. Two ponies injected intravenously with live E. sennetsu did no develop clinical illness. Subsequent injection with live E. sennetsu did not develop clinical illness. Subsequent injectio...
Blach EL, Amann RP, Bowen RA, Sawyer HR, Hermenet MJ.Transmission electron microscopy was used to confirm that a monoclonal antibody (F79.3E2; class IgG1 kappa) was specifically localized to an antigen in the acrosomal ground substance of stallion sperm. This antibody was used to develop and validate an indirect immunofluorescent procedure to evaluate integrity of the plasma-acrosomal membranes of stallion sperm. The concept was that primary monoclonal antibody would be "shielded" from its acrosomal antigen by an intact plasma membrane. Conversely, sperm with damaged plasma-acrosomal membranes would exhibit green acrosomal fluorescence when view...
How SJ, Lloyd DH, Lida J.A monoclonal antibody (McAb) to Dermatophilus congolensis was produced from murine hybridoma cultures and purified by affinity chromatography. Species specificity was demonstrated using indirect immunofluorescent staining; the McAb was shown to react with 10 D congolensis isolates but not with 10 Nocardia species isolates, a Rhodococcus and a Streptomyces species isolate. The McAb was used to demonstrate D congolensis in clinical material from confirmed bovine and ovine cases and presumptive equine cases of dermatophilosis by indirect immunofluorescent staining.
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 ...
Magnarelli LA, Anderson JF, Shaw E, Post JE, Palka FC.During 1982 and 1985, blood samples from 705 equids were examined for antibodies to Borrelia burgdorferi. By indirect immunofluorescence staining, IgM and total immunoglobulin (IgM and IgG) antibodies were detected in 37 (5.3%) and 90 (12.8%) serum specimens, respectively. The geometric mean titer for IgM antibody (140.4) was highest during July, whereas total immunoglobulin ranged from 94.1 in October to 338 in May. Eighty-six equids with total immunoglobulin to B burgdorferi lived in areas of Connecticut where the primary tick vector, Ixodes dammini, was present. Of the 86 equids, 9 from Lym...
Burgess EC.Blood samples from Wisconsin horses and cows suspected of having clinical disease due to Borrelia burgdorferi infection were submitted by veterinary practitioners. All serum, milk, colostrum, and synovial samples were tested for B. burgdorferi antibodies by immunofluorescence. Whole blood, milk, colostrum, and synovial fluid samples were cultured for B. burgdorferi. Records were kept on the clinical signs of antibody-positive animals, date of sample, and location of the animal by county. Of the samples tested for antibodies 282/430 cow sera, 118/190 horse sera, 5/10 cow synovial fluids, 3/6 ho...
Kuttler KL, Goff WL, Gipson CA, Blackburn BO.Both the complement-fixation test (CFT) and the indirect fluorescent antibody test (IFAT) were conducted on weekly serum samples from nine Arab geldings for 28 days before and 256 days after their exposure to Babesia equi of European origin. On an average the IFAT became positive 8 days before the CFT and showed higher relative serum titer increases. Both test procedures successfully detected infection and neither showed an appreciable drop in titer during this time frame, with the exception of the CFT, which showed a transient drop immediately following treatment with imidocarb. A test conduc...
White KL, Thomson DL, Wood TC.An indirect immunofluorescence assay was used to detect the presence of H-Y antigen on equine blastocysts. A total of 33 blastocyst stage horse embryos were collected 6 to 7 days post-ovulation by trans-cervical flush and were immediately evaluated for the presence of H-Y antigen. Additionally, 17 embryos, were collected and cultured for 72 h to the expanded blastocyst stage and similarly evaluated. Embryos were placed in medium containing monoclonal antibodies to H-Y antigen followed by incubation in medium containing 1/10 (v/v) fluorescein isothiocyanate conjugated goat anti-mouse IgM Fc spe...
Crump AL, Davis W, Antczak DF.A cell surface molecule of equine T lymphocytes was identified and characterized using a mouse monoclonal antibody, HT23A. The molecule was detected on all T cells but not on other cells in peripheral blood, with the possible exception of a small subpopulation (about 5%) of B cells, as assessed by indirect immunofluorescence and flow cytometry. HT23A labelled T cell areas of horse lymph nodes and spleen when used in an indirect immunoperoxidase assay on frozen sections. Macrophages and neutrophils were not labelled by the antibody nor were frozen sections of horse liver, kidney, or brain. HT23...
Wyatt CR, Magnuson NS, Perryman LE.Six monoclonal antibodies, designated EqT2, EqT3, EqT6, EqT7, EqT12, and EqT13, which identify T lymphocyte antigens present at different stages of T cell maturation were used to examine T lymphocyte development in foals with severe combined immunodeficiency (SCID). Flow microfluorimetry demonstrated the presence of EqT12+ and EqT13+ prothymocytes and a few phenotypically mature EqT2+ and EqT3+ thymocytes within the thymic remnants of SCID foals. However, very few EqT6+ and EqT7+ resident cortical thymocytes were detected. The near absence of EqT6+ and EqT7+ cortical thymocytes was confirmed b...
McEntee M.A cutaneous mass (1.5 cm in diameter) was removed from the head of a horse and was diagnosed histologically as eumycotic mycetoma. Immunofluorescence, performed on formalin-fixed, paraffin-embedded tissue, identified Pseudallescheria boydii as the etiologic agent. Findings from earlier reports of eumycotic mycetoma were compared with those of this horse.
de Waal DT, van Heerden J, Potgieter FT.Serologically negative horses, as determined with the indirect fluorescent antibody test (IFA), were infected with Babesia equi and 60 days later with Babesia caballi. The only clinical signs of disease observed in these animals were a febrile reaction and slight icterus. Haematological changes included a drop in haematocrit and haemoglobin concentration, as well as lowered platelet counts. The serum concentrations of albumin, iron and phosphorus were lowered. Mildly elevated serum bilirubin and fibrinogen concentrations were observed. Antibody titres were determined with the IFA and complemen...
Kitson-Piggot AW, Prescott JF.Sera from Thoroughbred and Standardbred horses in southwest Ontario were tested for antibody to seven Leptospira interrogans serovars (autumnalis, bratislava, canicola, grippotyphosa, hardjo, icterohaemorrhagiae, pomona), using the microscopic agglutination test. There was significantly higher seroprevalence of bratislava than of other serovars, in which prevalence was low. Seroprevalence of bratislava increased significantly with age; only 5% of two to three year old horses had titers greater than or equal to 1:80 compared to 52% of horses older than seven years. Eight of 16 foals from two fa...
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...
Morier L, Cantelar N, Soler M.Eastern Equine Encephalitis (EEE) was in Cuba before the 1940s; the virus has been isolated from horses, birds, and rodents during epizootic as well as interepizootic periods. The only isolation of Western Equine Encephalitis (WEE) virus was from a sick pigeon found in the vicinity of Havana University. Both viruses can cause human disease; the isolation of WEE virus from the centre of an urban area emphasises the need for the prompt isolation and rapid identification of these agents. The object of this work was to compare the sensitivity of a continuous cell line (XL-2) from the toad, Xenopus...
Bousquet D, Guillomot M, Betteridge KJ.The capsule which surrounds the pre-attachment equine embryo has been compared with the zona pellucida (zp) that it replaces, as well as with the rabbit blastocyst coverings, by means of physicochemical and immunological methods. Trypsin solution at pH varying between 7.5 and 9.0 completely solubilized the capsule, as did Na borohydride. However, solutions of pH 2.0 or 12.0, urea, high temperature (65 degrees C, 60 min or 80 degrees C, 30 min), mercaptoethanol and dithiothreitol were able to solubilize the zp but not the capsule at the concentrations used. Indirect immunofluorescence on cryost...
Sentsui H, Kono Y.Horse erythrocytes treated with equine infectious anemia virus hemagglutinin were found to be lysed after incubation with fresh horse serum at 37 degrees C. Fresh guinea pig serum induced more efficient hemolysis than horse serum. Direct immunofluorescence test revealed the adsorption of complement factors on the surface of the erythrocytes. Calcium and magnesium ions were necessary for the hemolysis to take place. Antibody against equine infectious anemia virus enhanced the virus-induced complement-mediated hemolysis. These observations indicated that the classical pathway of complement activ...
Kuttler KL, Gipson CA, Goff WL, Johnson LW.Nine 4-year-old Arabian geldings were experimentally infected with Babesia equi of European origin. All horses developed detectable parasitemia an average of 30 days after they were inoculated, which was accompanied by a decrease in PCV. The infections were generally mild with no animal deaths. All horses became serologically positive by the indirect fluorescent antibody test within an average of 23 days after they were inoculated and by the complement-fixation test 30 days after they were inoculated.
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...
Winder NC, von Fellenberg R.Pulmonary parenchymal tissues from 6 healthy horses and from 9 horses with chronic bronchiolitis were evaluated by use of an indirect immunofluorescent technique. In horses of both groups, the diffuse interstitial immunofluorescence was most intense for immunoglobulin (Ig)G, moderate for IgA, and minimal for IgM. Intensity of fluorescence was increased markedly in diseased lungs exposed to anti-IgA and anti-IgG. Around small bronchi and large bronchioles of healthy horses, IgA-containing cells generally were more numerous than were IgG-Fc fragment (Fc)-containing cells; in small bronchioles, h...
Ishak GM, Dutra GA, Gastal GDA, Elcombe ME, Gastal MO, Park SB, Feugang JM, Gastal EL.This study aimed to gain insight on the effect of different seasons of the year on the expression pattern of growth factor and hormone receptors involved in follicle development. A novel follicle wall biopsy technique was used to collect in vivo follicle wall layers (ie, granulosa, theca interna, and theca externa) and follicular fluid samples from growing dominant follicles, simultaneously and repeatedly, using the same mares during the spring anovulatory (SAN), spring ovulatory (SOV), summer (SU), and fall ovulatory (FOV) seasons. The immunofluorescent expression patterns of epidermal growt...
Abu-Halaweh M, Abo-Shehada MN, Khalil R.A cross-sectional study was carried out on a sample of 379 horses to determine the seroprevalence of Neospora spp. in Jordan using the indirect fluorescent antibody test. Five variables, namely locality (n=10), climatic zone (n=4), age group (n=3), gender, and breed were tested as risk factors for Neospora-immunoglobulin (Ig)G seropositivity at four cutoff titers (1:50, 1:200, 1:400, and 1:800) using univariate and multivariate logistic regression analyses. A total of 122 (32%; 95% CI: 28, 37) sera samples had anti-Neospora-IgG at a cutoff titer of 1:50. Increased Neospora-IgG seropositivity w...
Whyte A, Ockleford CD, Wooding FB, Hamon M, Allen WR, Kellie S.The establishment of a monolayer culture of cells derived from the girdle region of a 34-day-old donkey conceptus is described. These cells have had over 100 repeated passages in culture. Low levels of pregnant mares' serum gonadotrophin (PMSG, eCG) could be detected in the cells by indirect immunofluorescence using some monoclonal anti-eCG antibodies, but the cells did not secrete eCG as measured by radioimmunoassay or inhibition of haemagglutination. There was marked nuclear polymorphism with binucleate and occasional multinucleate cells. The cells were strongly reactive with wheatgerm agglu...
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...
Mulville P.In the late 1970s, a new infectious disease in horses, involving acute enteritis, was recognised in the Potomac River area of Maryland, U.S.A. The causative agent was identified subsequently as a new species of rickettsial organism, later named Ehrlichia risticii. Since then, the disease has been reported in many other states, and in enzootic areas vaccination is common. Signs associated with the clinical disease included depression, fever, anorexia, decreased or absent intestinal sounds, profuse watery diarrhoea and laminitis. However, considerable variation in clinical manifestations has bee...
Winfield LD, White SD, Affolter VK, Renier AC, Dawson D, Olivry T, Outerbridge CA, Wang YH, Iyori K, Nishifuji K.To describe the clinical, histological and immunological findings of an equine case of pemphigus vulgaris, including the demonstration of antidesmoglein (anti-Dsg) autoantibodies. Methods: The diagnosis of pemphigus vulgaris was confirmed in a 9-year-old Welsh pony stallion with both direct and indirect immunofluorescence and immunoprecipitation studies, the latter identifying circulating anti-Dsg3 IgG. Treatment with immunosuppressive medications was initiated. Lesions were seen in the perineal area, sheath, mane, tail, eyelids, coronary bands and mucosa of the mouth and oesophagus. Initial c...
Baldwin WM, Armstrong LP, Samaniego-Picota M, Rahimi S, Zachary AA, Kasper EK, Conte JV, Hruban RH, Rodriguez ER.Polyclonal antithymocyte globulin preparations contain antibodies with reactivity to endothelial cells. Therefore, we investigated whether treatment with this reagent caused complement deposition in human cardiac transplants. Frozen tissue was available from endomyocardial biopsies of 75 patients, who were transplanted between April 1995 and April 2000. Nine of these patients were converted from cyclosporin A (CsA) to horse antithymocyte globulin (ATGAM) in the first month after transplantation. All of the biopsies were stained by immunofluorescence for C4d as evidence of activation of the cla...
Truax RE, Powell MD, Montelaro RC, Issel CJ, Newman MJ.A rapid and simple technique for the cryopreservation and recovery of equine mononuclear cells was developed. Buffy-coat leukocytes were frozen in autologous plasma containing 10% DMSO and mononuclear cells were recovered by gradient sedimentation using a standard Ficoll-Hypaque purification procedure. The total numbers of mononuclear cells recovered from cryopreserved samples were 94%-82% of those recovered from fresh blood samples. The functional capabilities of the mononuclear cells from cryopreserved buffy coat preparations were compared with those of mononuclear cells from fresh samples b...
Barbis DP, Bainbridge D, Crump AL, Zhang CH, Antczak DF.A panel of monoclonal antibodies was used to characterize the expression of equine Major Histocompatibility Complex (MHC) class II antigens on lymphocytes of horses of different MHC types. MHC class II antigen expression was compared between adult horses and foals, and the level of expression of MHC class II antigens on horse T cell subpopulations was also determined. Peripheral blood lymphocytes (PBL) from young and adult healthy horses of different MHC haplotypes were labeled with the antibodies and assayed by single- and two-color immunofluorescence flow cytometry. A variation in the expres...
Magnarelli LA, Van Andel AE, Ijdo JW, Heimer R, Fikrig E.To diagnose granulocytic ehrlichiosis in horses, compare results of indirect fluorescent antibody (IFA) staining procedures with those of immunoblot analysis, and compare serologic test findings with polymerase chain reaction (PCR) results. Methods: 69 horses with high rectal temperatures (> or = 39 C) and lethargy, anorexia, or limb edema. Methods: 43 convalescent serum samples obtained from 38 horses 2 to 18 weeks after onset of illness were analyzed by use of immunoblot procedures and IFA staining methods, using the NCH-1 or BDS ehrlichial strains. Blood samples from 69 acutely ill horse...
de Moura AB, da Silva MO, Farias JA, Vieira-Neto A, de Souza AP, Sartor AA, Fonteque JH, Bunn S.The aims of this study were to determine occurrences of Neospora spp. IgG antibodies in horses from two geographical regions of the state of Santa Catarina, southern Brazil, and identify risk factors for infection. Analyses were performed on 615 samples using the immunofluorescent antibody test (IFAT ≥ 1:50). Out of the 615 samples, 25 (4.1%) were positive for Neospora spp. The titers for Neospora spp. were distributed as follows: 1:50 (13), 1:100 (eight), 1:200 (three) and 1:400 (one). Out of the 311 samples taken in the mountain region, eight were positive (2.6%). Among the samples from th...
Smith HL, Berglund AK, Robertson JB, Schnabel LV, McMullen RJ, Gilger BC, Oh A.The objective of the study was to determine the effect of gentamicin on CD3+ T-lymphocyte proliferation and cell viability using an in vitro cell culture model as a means of investigating the mechanism of action of low-dose intravitreal gentamicin injection. Methods: Three adult horses with no evidence of ophthalmic or systemic disease. Methods: Peripheral blood lymphocytes were treated with gentamicin at concentrations 37.5 μg/mL, 112.5 μg/mL, 187 μg/mL, 375 μg/mL, or 750 μg/mL then stimulated to proliferate with concanavalin A (ConA). 4',6-diamidino-2-phenylindole (DAPI) and c...
Berman N, Tirosh-Levy S, Steinman A, Minderigiu A, Blinder E, Leszkowicz Mazuz M. is a tissue cyst forming coccidia, which affects multiple host species worldwide. Equine besnoitiosis is characterized mainly by generalized skin lesions and cysts in the scleral conjunctiva. Recent reports revealed exposure to in equines in Europe and the United States. However, the exposure to spp. in the Israeli equine population was never investigated. The aim of this study was to evaluate the seroprevalence and associated risk factors for besnoitiosis in equids in Israel. A cross-sectional serosurvey was performed using serum samples of apparently healthy horses ( = 347), donkeys ( = 9...
Lambertini C, Zannoni A, Romagnoli N, Bombardi C, Morini M, Dondi F, Bernardini C, Forni M, Rinnovati R, Spadari A.Large colon volvulus in horses is associated with a poor prognosis, especially when ischemic-reperfusion injury of the affected intestinal tract develops. Proteinase-activated receptor 2 (PAR2) plays an important role in the pathogenesis of inflammation in the gastrointestinal tract. The aim of this study was to evaluate the distribution and expression of PAR2 in colonic pelvic flexure of horses spontaneously affected by large colon volvulus (CVH group). Eight horses admitted for severe abdominal colon volvolus and which underwent surgery were included. Colon samples were collected after enter...
van de Moer A, Rice M, Wilks CR.A type-specific monoclonal antibody was produced by immunizing mice with purified equid herpesvirus-1 (EHV-1). The EHV-1 specific mAb reacted with all the EHV-1 strains tested so far by indirect ELISA, immunofluorescence, and immunoperoxidase tests. No reactions were detected with the EHV-4, EHV-2, or EHV-3 strains tested. The indirect immunofluorescence and immunoperoxidase tests showed that the nuclei of infected cells were predominantly stained by this mAb. Triton treatment of the virus and immunogold labeling experiments indicated that the nucleocapsid of EHV-1 was the target antigen of th...
Minervino AHH, Andersson E, Norlander E, Moreira TR, Gennari SM, Mercado Caruso N, Moraes Filho J, Marcili A, Martins TF, Labruna MB.From a previous large epidemiological survey, we randomly selected 474 serum samples (463 horses and 11 mules) distributed among four municipalities of Pará state, Amazon region, Brazil, and from three types: farm animal, urban carthorse, and sport horse. Samples were tested by indirect fluorescence antibody test (IFAT ≥ 64) for antibodies reactive to spotted fever group (SFG) rickettsiae using Rickettsia rickettsii as crude antigens. From the 474 equids tested, 149 (31.4%) had ticks attached during sampling, belonging mostly to the species Dermacentor nitens. The overall seroprevalence...
Zeng W, Alpaugh W, Stefanovski D, Schlingmann K, Dobrinski I, Turner RM.The study of spermatogenesis in the horse is challenging because of the absence of an in vitro system that is capable of reproducing efficient spermatogenesis and because of the difficulties and costs associated with performing well-controlled studies in vivo. In an attempt to develop novel methods for the study of equine spermatogenesis, we tested whether cells from enzymatically digested pre-pubertal equine testicular tissue were capable of de novo tissue formation and spermatogenesis following xenografting under the back skin of immunocompromised mice. Testes were obtained from normal pre...
Rehbein G, Heidrich-Joswig S.Eight ponies infected with Babesia equi were investigated for their serological response to B. equi schizont and piroplasm antigen with the indirect fluorescent antibody test (IFAT) and complement fixation test (CFT). Piroplasm antigen was prepared from an infected splenectomized pony, while schizont antigen was produced from cultured lymphoid cells which contained B. equi macroschizonts. The IFAT detected a rise in antibody titres to schizont antigen as well as to piroplasm antigen, but differences were obtained in the duration of antibody detection. Significant antibody titres to piroplasm a...
Reinert M, Calvete JJ, Sanz L, Töpfer-Petersen E.SSP-7 is a protein originally isolated from stallion seminal plasma. It has extensive amino acid sequence homology with boar spermadhesin AWN, and, like its porcine counterpart, SSP-7 displays zona pellucida-binding activity. Strikingly, however, immunohistochemical studies presented here show that the stallion and the boar spermadhesin homologues are secreted at different places of the male genital tract. Furthermore, indirect immunofluorescence shows that the topography of SSP-7 on the surface of stallion spermatozoa is restricted to the equatorial segment, whereas boar AWN epitopes cover th...
Yamaguchi K, Sawada T, Yamane S, Haga S, Ikeda K, Igata-Yi R, Yoshiki K, Matsuoka M, Okabe H, Horii Y, Nawa Y, Waltrip RW, Carbone KM.Borna disease virus (BDV) is a neurotropic pathogen that infects a wide variety of vertebrates. We have developed a new electrochemiluminescence immunoassay (ECLIA) for the detection of antibodies to BDV, using three synthetic peptides corresponding to the amino acid residues 3-20 and 338-358 of p40 and 59-79 of p24 peptide of BDV. Using the ECLIA, we examined serum samples for the presence of anti-BDV antibodies in 20 rats (experimentally BDV-infected and uninfected) and 38 horses (13 US horses, experimentally infected and uninfected, and 25 Japanese horses, feral and domestic). The ECLIA, pe...
Nedjari M.Investigations to occurrence of Sarcocystiosis of domestic animals in Algeria were carried out by different methods--Immunofluorescence (IFAT), macroscopic and microscopic examinations of muscle, coproscopia. Positive results could be found in intermediate hosts--362 of 537 cattle, 188 of 292 sheep, 13 of 57 horses and in the final hosts dogs and cats. Muscle-probes from cattle contained S. cruzi (58.7 respectively 61.0%), S. hirsuta (48.9 respectively 35.0%), from sheep S. tenella (61.0 respectively 59.6%, S. arieticanis (38.8 respectively 40.3%), from horses S. bertrami (24.5%). IFAT-data of...
Pazinato FM, Curcio BDR, Varela Junior AS, Corcini CD, Wendt CG, Moreira F, Schmit RA, Nogueira CEW.Placental tissues from humans, rodents, and farm animal contain leptin and its receptor. Expression of leptin has already been described in horses, although there is no description about immunolocalization of leptin and its receptor. The aim of the present study was to investigate the presence and distribution of leptin and ObR-b in the equine placenta at term by immunofluorescence staining, and the changes on plasma leptin concentrations during late gestation. The present study involved eight Criollo-type mares carrying healthy pregnancies. Blood samples were collected during the third trimes...
Chodkowski M, Słońska A, Gregorczyk-Zboroch K, Nowak-Zyczynska Z, Golke A, Krzyżowska M, Bańbura MW, Cymerys J.Mitochondria are key cellular organelles responsible for many essential functions, including ATP production, ion homeostasis and apoptosis induction. Recent studies indicate their significant role during viral infection. In the present study, we examined the effects of equine herpesvirus type 1 (EHV-1) infection on the morphology and mitochondrial function in primary murine neurons in vitro. We used three EHV-1 strains: two non-neuropathogenic (Jan-E and Rac-H) and one neuropathogenic (EHV-1 26). The organization of the mitochondrial network during EHV-1 infection was assessed by immunofluores...
Harkiss GD, Brown DL, Smith DJ, Nagington J.Circulating immune complexes were isolated from the sera of cardiac allograft recipients by bovine conglutinin/anti-conglutinin co-precipitation, or by gel filtration and protein A-Sepharose affinity chromatography. The antibody moieties within these isolated immune complexes were tested for specificity against heterologous anti-thymocyte globulins by solid phase radioimmunoassay, and bacterial and viral antigens by indirect immunofluorescence. The results showed that in addition to possessing specific anti-equine anti-thymocyte globulin antibodies, immune complexes also contained cross-reacti...
Chen J, Guo X, Li L.The nucleocapsid (N) protein is the most conserved structural protein in equine arteritis virus (EAV). This study aimed to identify the minimal conserved B cell epitope on the EAV N protein. The purified N protein was used to immunize mice for preparing monoclonal antibody (mAb). The reactivity of mAb was evaluated by Western blot and immunofluorescence assay. Moreover, 11 overlapping peptides (named MBP-N1 to MBP-N11) were designed to localize the linear antigenic epitope within the N protein. The peptides were identified by indirect enzyme-linked immunosorbent assay (ELISA) and Western blot....
Strazzullo M, Corteggio A, Altamura G, Francioso R, Roperto F, D'Esposito M, Borzacchiello G.Sarcoids are peculiar equine benign tumours. Their onset is associated with Bovine Papillomavirus type -1 or -2 (BPV-1/2) infection. Little is known about the molecular interplay between viral infection and neoplastic transformation. The data regarding papillomavirus infections in human species show the inactivation of a number of tumour suppressor genes as basic mechanism of transformation. In this study the putative role of the tumour suppressor gene Fragile Histidine Triad (FHIT) in sarcoid tumour was investigated in different experimental models. The expression of the oncosuppressor protei...
Sellon DC, Cullen JM, Whetter LE, Gebhard DH, Coggins L, Fuller FJ.An IgG1 mouse monoclonal antibody, designated 1.646, is described which recognizes a cytoplasmic antigen of equine mononuclear phagocytes. Indirect fluorescent antibody staining of peripheral blood leukocytes reveals a granular cytoplasmic staining, predominantly in adherent blood mononuclear cells. Indirect fluorescent antibody staining is positive for alveolar and peritoneal macrophages. In some horses, a few neutrophils are also stained. In equine tissue samples stained by immunohistochemistry, the distribution of positive cells is consistent with the distribution of tissue macrophages. The...
Smith IM, Girard A, Corner AH, Mitchell D.Using two known positive equine viral rhinopneumonitis (EVR) sera, conjugates were prepared with fluorescein isothiocyanate and tested for specificity using EVR infected tissue culture cells. The conjugate was then applied to selected tissues from 32 aborted fetuses and foals submitted during a natural outbreak of EVR. Antigen was detected in various tissues by immunofluorescence in 20 cases (62.5%). In 24 cases bovine fetal kidney cell monolayers were inoculated with a pool of lung and liver and EVR virus was isolated from 15 (62.5%). Histological examination of various tissues from 29 cases ...
Eizema K, van der Wal DE, van den Burg MM, de Jonge HW, Everts ME.During early postnatal development, the myosin heavy chain (MyHC) expression pattern in equine gluteus medius muscle shows adaptation to movement and load,resulting in a decrease in the number of fast MyHC fibers and an increase in the number of slow MyHC fibers. In the present study we correlated the expression of MyHC isoforms to the expression of sarcoplasmic(endo)reticulum Ca2+-ATPase 1 and 2a (SERCA), phospholamban (PLB), calcineurin A (CnA), and calcineurin B (CnB). Gluteus medius muscle biopsies were taken at 0, 2, 4, and 48 weeks and analyzed using immunofluorescence. Both SERCA isofor...
Sevcik C, D'Suze G, Salazar V, Díaz P, Vázquez H.We used high sensitivity and resolution fluorescence microscopy to study the interaction of ostrich IgY, horse F(ab')₂ and horse IgG with mice lymphocyte and erythrocyte plasma membrane. The immunoglobulins were labeled with fluorescein isotiocyanate (FITC). Our results show an interaction of IgY with lymphocyte plasma membrane which does not result in endocytosis of the labeled protein. Less IgG and its F(ab')₂ fraction bind to lymphocytes, and this binding seems to be followed by endocytosis producing a diffuse cytoplasmic fluorescence in most lymphocytes exposed to FITC-IgG or FITC-F(ab...
