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.
Granstrom DE, Dubey JP, Davis SW, Fayer R, Fox JC, Poonacha KB, Giles RC, Comer PF.Antigens of cultured Sarcocystis neurona merozoites were examined using immunoblot analysis. Blotted proteins were probed with S. cruzi, S. muris, and S. neurona antisera produced in rabbits, S. fayeri (pre- and post-infection) and S. neurona (pre- and post-inoculation) sera produced in horses, immune sera from 7 histologically confirmed cases of equine protozoal myeloencephalitis (EPM), and pre-suckle serum from a newborn foal. Eight proteins, 70, 24, 23.5, 22.5, 13, 11, 10.5, and 10 Kd, were detected only by S. neurona antiserum and/or immune serum from EPM-affected horses. Equine sera were ...
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...
Pelegrino JL, Vázquez S, Morier L, Castillo A, Guzmán MG, Kourí G.We present the results attained in the identification of Eastern equine encephalomyelitis virus isolations in Vero and XL-2 cell systems, using a double-antibody ELISA technique and the indirect immunofluorescence method. The results attained through these two techniques coincided by 100% with identification through neutralization. With the former, the virus was detected within 6-8 hours after inoculation. Better results were attained with XL-2 cells.
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...
Ranz AI, Miguet JG, Anaya C, Venteo A, Cortés E, Vela C, Sanz A.A panel of 32 hybridoma cell lines secreting monoclonal antibodies (MAbs) reactive with African horsesickness virus serotype 4 (AHSV-4) has been developed. Four of the MAbs recognized the major core antigen VP7, twenty recognized the outer capsid protein VP2 and eight reacted with the non-structural protein NS1. With the VP7-specific MAbs a rapid and sensitive double antibody sandwich immunoassay has been developed to detect viral antigen in infected Vero cells and in spleen tissue from AHSV-infected horses. The sensitivity of the assay is 10 ng viral antigen per 100 microliters. The NS1-speci...
Böse R, Daemen K.Babesia caballi-infected or normal equine erythrocytes were solubilized in sodium dodecyl sulfate (SDS) buffer and analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Antigens were allowed to react with sera from horses experimentally or field-infected with B. caballi and with sera from non-infected horses. Major babesial antigens recognized by immune sera had apparent mol. wts of 141, 112, 70, 50, 48, 34, and 30 kDa. The polypeptides at 50 and 48 kDa were recognized earliest and throughout infection, but also weakly by 3/100 equine sera tested negative and 1/33...
Vittoria A, Cocca T, La Mura E, Cecio A.The aim of this study is to describe the presence of neuroendocrine (NE) cells (paraneurons), producing biogenic amines and/or peptidergic hormones, in the female urethra of cattle, sheep, pigs, and horses, by means of histochemical and double labeling immunofluorescent techniques. 5-Hydroxy-tryptamine-, chromogranin A-, cholecystokinin- and somatostatin-containing NE cells are present in the urethral epithelium of all the species studied, with the unique exception of the lack of somatostatin cells in the horse. Paraneurons containing 5-hydroxytryptamine colocalized with chromogranin A or chol...
Divers TJ, Timoney JF, Lewis RM, Smith CA.A 12-year-old thoroughbred gelding died from diffuse global glomerulonephritis, 3 months after a lower respiratory infection from which Streptococcus zooepidemicus was isolated. Immunopathological studies (immunofluorescence, immunodiffusion, immunoperoxidase testing and immunoblotting) indicated the presence of an immune reactant renal disease associated with IgG antibody and streptococcal antigens.
Pospischil A, Lieb A, Corboz L.In Switzerland during the foaling season 1988 and 1989 the cause of abortion in 60 foals was investigated. Special attention was paid to infections with equine herpesvirus 1 (EHV 1). Diagnosis were based on post-mortem, histopathological, bacteriological and immunofluorescence investigation. The results confirm data from other countries, that EHV 1 is the most prevalent viral (20%) cause of abortion, followed by various bacterial agents (12%). Other causes were umbilical torsion, twin pregnancy and malformations. In 18% of the cases the investigation of fetuses did not give any results as to t...
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...
Kitamoto N, Ramig RF, Matson DO, Estes MK.The production of viral antigen after infection of MA104, HepG2 (derived from human liver), and CaCo-2 (derived from human colon) cells with various cultivatable human and animal rotavirus strains was compared using immunofluorescence tests. All rotavirus strains examined expressed antigen in CaCo-2 cells and MA104 cells, but only some virus strains, namely, SA11-Cl3 (simian), RRV (simian), CU-1 (canine), and Ty1 (turkey), produced antigen in numbers of infected HepG2 cells comparable to infections in MA104 and CaCo-2 cells. Fl-14 (equine), OSU (porcine), NCDV (bovine), and Ch2 (chicken) strai...
Lunn DP, Holmes MA, Duffus WP.The aim of this study was to produce monoclonal antibodies (mAb) recognizing equine lymphocyte surface antigens. Fusions were conducted using BALB/c mice hyperimmunized with equine thymocytes. Hybridoma supernatants were screened by flow cytometry and positive hybridomas were cloned twice by limiting dilution. These mAb were then characterized for tissue distribution by immunohistology and flow cytometry, and by precipitation and analysis of the lymphocyte antigens which they recognized. Three mAb (CVS5, CVS4 and CVS8) are described which recognize only T lymphocytes in peripheral blood. Two-c...
Edington N, Smyth B, Griffiths L.One of three mares in the last trimester of pregnancy became paraplegic 7 days after experimental infection with EHV-1 and was killed 10 days after infection (d.p.i.). The other two mares aborted foetuses at 12 and 14 d.p.i. In the first mare, virus was detected by immunofluorescence (IIF) and immunoperoxidase (IP) staining in endothelial cells of the endometrium, placenta and umbilical vein, but not in any other foetal tissues. In the experimentally aborted foetuses, and in two other independent field cases of abortions, endothelial cell infection was also detected in the foetuses, both in ma...
Gaĭdamovich SIa, Pomelova VG, Lavrova NA, Mel'nikova EE, Sokolova MV, Kharitonenkov IG, Zlobin VN.Potentialities of differentiation between Venezuelan equine encephalomyelitis (VEE) complex viruses by time-resolved fluoroimmunoassay and enzyme immunoassay were studied. For this, 4 test systems were used based on different combinations of native and labeled polyclonal antibodies to VEE virus, strain Trinidad, and monoclonal (MCA) antibody MAK 14-7 to protein EL of this virus. The maximal sensitivity and specificity was achieved in the test system formed from native MCA MAK 14-7 for sensitization of the solid phase and labeled polyclonal immunoglobulins for demonstration of the test results....
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...
Grabner A, Fischer A.In a retrospective study of 38 horses with Borna encephalitis which were clinically and histopathologically examined in the "I. Medizinische Tierklinik" in Munich between 1977 and 1990, the epidemiology, the clinical symptomatic and the diagnostic procedures available are presented. Indirect immunofluorescence showed antibodies in the serum of 12 out of 29 cases (41%) and in the cerebrospinal fluid (CSF) of 17 out of 28 cases (61%). The evaluation of 23 cases in which indirect immunofluorescence of serum and CSF, and also the post mortem virological and histopathological examination of the bra...
van der Kolk JH, Bernadina WE, Visser IJ.A four year old Dutch warmblooded mare was born and raised in the province of North-Brabant, the Netherlands. On May 16, 1989, she showed signs of colic, anorexia, depression, ileus, severe dehydration and leukopenia. When the mare collapsed, euthanasia was carried out. Acute colitis and cytoplasmic inclusion bodies in macrophages were observed at autopsy. When an indirect immunofluorescence assay was performed, the Ehrlichia risticii titre of the serum was found to be 1:640.
Käsbohrer A, Schönberg A.The prevalence of B. burgdorferi, the causative agent of Lyme Borreliosis in humans, was determined in domestic animals living in Berlin. 189 dogs, 29 cats, 224 horses and 194 cows were investigated. Using the indirect immunofluorescence test (IFT) 5.8% of the dogs and 24.5% of the cows investigated showed a positive reaction at titres of 1:128 or higher. Horses and cats gave negative results. ELISA was more sensitive than IFT. 10.1% of the dogs, 16.1% of the horses and 66% of the local cows showed positive reaction. Domestic animals seem to be in contact with B. burgdorferi and can be a reser...
Zhang J, Boyle MS, Smith CA, Moore HD.The acrosome of the stallion spermatozoon was visualized by indirect immunofluorescence with monoclonal antibody (18.6) which recognized an integral acrosomal membrane component. Localization was confirmed by electron microscopy using peroxidase labelled antibody. In fresh semen samples (n = 19), 73.9 +/- 9.1% of the spermatozoa from five fertile stallions displayed a uniform bright fluorescence over their acrosome region. In two semen samples from an infertile stallion only 28% and 35% of spermatozoa showed the same pattern of fluorescence. Spermatozoa from fertile stallions incubated for up ...
Bochsler PN, Slauson DO, Neilsen NR.The essential role of the CD11/CD18 family of leukocyte adhesion molecules (LeuCams) in neutrophil-substrate adhesion is well documented. We have found that a monoclonal antibody designated 60.3 (MoAb 60.3) that recognizes the common beta-subunit (CD18) on human neutrophils (PMN) also recognizes a surface antigen on equine PMN. Antigen expression as assessed by immunofluorescence flow cytometry was enhanced by zymosan-activated serum (ZAS) or phorbol 12-myristate 13-acetate (PMA) stimulation. Pretreatment of equine PMN with MoAb 60.3 inhibited ZAS-stimulated aggregation, indicating that the mo...
Jain NC, Vegad JL, Kono CS.Equine neutrophil antibody was raised in rabbits inoculated with equine neutrophils isolated to purity greater than 99.0%, using Percoll density-gradient sedimentation. Neutrophil antibody was detected by use of agar gel diffusion, leukoagglutination, indirect immunofluorescence, staphylococcal protein A and streptococcal protein G binding, and phagocytic inhibition techniques. Precipitin lines and leukoagglutination were seen in antiserum dilutions of 1:4 and 1:64, respectively. The specific nature of leukoagglutination was characterized by the formation of rosette-like clumps of neutrophils....
Wills JM, Watson G, Lusher M, Mair TS, Wood D, Richmond SJ.This paper describes the isolation and characterisation of a strain of Chlamydia psittaci obtained from a nasal swab taken from a horse with serous nasal discharge. Initial isolation was achieved in cycloheximide-treated McCoy cell monolayers. Chlamydial inclusions stained by immunofluorescence either with a rabbit antiserum raised against C. psittaci or with a monoclonal antibody directed against the genus-specific lipopolysaccharide antigen were single and compact. They did not stain with iodine or with a monoclonal antibody reactive against Chlamydia trachomatis. The agent was re-isolated i...
Anestad G, Maagaard O.During an epizootic of equine influenza in Norway caused by influenza A/equine (H3N8) virus the efficacy of rapid virus diagnosis by the indirect immunofluorescence technique was evaluated. The antiserum used in the test was a polyclonal influenza A virus antiserum with reactivity directed mainly against the common nucleoprotein and matrix protein. This antiserum possessed sufficient reactivity for the detection of virus-infected exfoliated nasopharyngeal cells. Nasopharyngeal smear samples from 92 horses were examined and a positive diagnosis was obtained for 57 (62 per cent). Paired serum sa...
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...
Bell CW, Boyle DB, Whalley JM.Transcript mapping of the equine herpesvirus 1 (EHV-1) glycoprotein B (gB) gene homologue by Northern blot, S1 nuclease and primer extension analyses indicated that two overlapping transcripts of 3.4 and 4.6 kb originated from the same strand and were transcribed from left to right between coordinates 0.40 and 0.43 of the EHV-1 genome. The 3.4 kb transcript encoded EHV-1 gB and the 5' RNA terminus was located approximately 30 bases downstream from a probable TATA element. The coding region of the gB gene homologue was reconstructed from two subclones using oligonucleotide mutagenesis and inser...
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...
van den Heuvel LP, van den Born J, Veerkamp JH, Janssen GH, van de Velden TJ, Monnens LA, Schröder CH, Berden JH.1. Proteoglycans extracted from human and equine glomerular basement membranes (GBM) were purified by ion-exchange chromatography and gel filtration. 2. The glycoconjugates had an apparent molecular mass of 200-400 kDa and consisted of 75% protein and 25% glycosaminoglycan. Glycosidase and HNO2 treatment and the amino sugar and sulfate composition of both proteoglycan preparations identified heparan sulfate (HS) as the predominant saccharide chain. 3. Hydrolysis with trifluoromethanesulfonic acid yielded comparable core proteins with molecular masses of ca 160 and 120 kDa. 4. The HS chains had...
Goetz TE, Holland CJ, Dawson JE, Ristic M, Skibbe K, Keegan KG, Johnson PJ, Schaeffer DJ, Baker GJ.The seroprevalence and seasonal trend of antibody titers against equine monocytic ehrlichiosis (Potomac horse fever) were determined in apparently healthy horses in selected areas of Illinois in 1986. Sera from 1,367 horses (6 months to 29 years old) were evaluated for the presence of antibodies against Ehrlichia risticii with indirect immunofluorescence. The majority (88%) of the horses were Thoroughbred or Standardbred racehorses. The number of horses with antibodies against E risticii was 229/1,367 (16.75%). The titers in these horses ranged from 1:10 to 1:640. As the year progressed, the n...
Dutta SK, Mattingly BL, Shankarappa B.The antibody response and the antibody reactivity to component antigens of Ehrlichia risticii were studied in horses with induced Potomac horse fever. These horses had no detectable antibodies to E. risticii in their preinoculation (PrI) sera by indirect fluorescent-antibody assay and enzyme-linked immunosorbent assay (ELISA). All the horses exhibited typical disease features following experimental infection and responded with specific antibodies, as measured by ELISA and indirect fluorescent-antibody assay. A primary antibody response was detected in 70% of the horses, while a secondary-type ...
Williams MA, Dowling PM, Angarano DW, Yu AA, DiFranco BJ, Lenz SD, Anhalt GJ.An adult horse with a 2-month history of anorexia, ataxia, and oral blisters had developed these clinical signs just prior to the appearance and growth of a cervical mass. Bullous stomatitis was characterized histologically as subepidermal clefting. Clinical signs were unresponsive to treatment with antibiotics or corticosteroids; however, surgical removal of the mass coincided with remission of all signs. Histologic findings of the mass were consistent with hemangiosarcoma. Results of indirect immunofluorescence and immunoprecipitation on frozen serum from the horse were characteristic of par...
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 ...
Granstrom DE.This article reviews recent advances in laboratory diagnosis of equine parasitic diseases. Laboratory diagnosis of most equine parasitic diseases continues to rely on standard methods. Only laboratory diagnostic tests for EPM, cryptosporidiosis, and giardiasis were included. The criteria for testing and interpretation of results for each new diagnostic method were explained. Western blot and PCR testing for EPM and immunofluorescent staining with monoclonal antibodies for cryptosporidiosis and giardiasis were reviewed.
Tessler J.The fluorescent antibody reaction was studied in tissues of ponies infected with African horsesickness virus (AHSV). Lung, spleen, lymph node, liver, skeletal muscle, intestine, stomach, nerve ganglion and kidney were sectioned and stained by the direct fluorescent antibody technique (FA). Fluorescence was demonstrated only in the spleen and could be inhibited by using unconjugated antiserum.
Magni E, Bertelloni F, Sgorbini M, Ebani VV.To investigate the occurrence of Bartonella sp. infection in asymptomatic horses and donkeys living in Tuscany, Central Italy. Methods: Blood samples were collected from 77 horses and 15 donkeys and tested by indirect immunofluorescent test to detect antibodies against Bartonella sp. and by PCR to detect the pathogen. Results: Fifty-four (58.69%; 95% CI: 47.95%-68.87%) animals, 9 donkeys and 45 horses, were seropositive with antibody titers ranging from 1:64 to 1:512. PCR assays detected 9 horses positive for Bartonella sp. and 3 donkeys for Bartonella henselae genotype I. Conclusions: The det...
The Journal of hygieneAugust 1, 1981
Volume 87, Issue 1 93-100 doi: 10.1017/s0022172400069278
Lemcke RM, Ernø H, Gupta U.Two unidentified mycoplasmas, N3 and N11, isolated from the respiratory tract of horses, were found to cross-react with strains of M. mycoides subsp. mycoides in indirect immunofluorescence tests, growth-inhibition tests carried out by the running drop/agar-well method, and in complement-fixation and double immunodiffusion tests. Serologically, the equine mycoplasmas were not completely identical with any of the reference strains of M. mycoides with which they were compared. Their cultural characteristics, ability to digest coagulated serum and casein, and survival at 45 degrees C, however, su...
Diethelm AG, Chambers LM, Balch CM, Phillips SJ.Anti-human thymocyte cell membrane antibody prepared by hyperimmunization of the horse produced an antiserum capable of prolonging skin allografts in the rhesus monkey for an average of 26 days. Lymphocyte depletion was present in paracortical areas of mesenteric lymph nodes of these animals after 28 days of treatment; the intravenous administration was tolerated without ill effects. Immunofluorescent studies identified both broad specificity antibodies reacting with numerous human cell types as well as thymus-dependent (T) cell antibodies reactive with human thymocytes and peripheral T-cells....
Edington N, Wright JA, Patel JR, Edwards GB, Griffiths L.Equine adenovirus 1 was recovered after four to six passages from two out of three cases of cauda equina neuritis (CEN) using kidney monolayers. Similar treatment of lumbo-sacral spinal cord from six normal horses did not yield adenovirus. All three cases of CEN had antibodies to the neuritogenic myelin protein P2 while immunofluorescence demonstrated that autologous IgG bound to the myelin of affected nerves. Adenovirus was not detected in neural tissue by immunofluorescence.
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.
Bernardino PN, Pusterla N, Conrad PA, Packham AE, Tamez-Trevino E, Aleman M, James K, Smith WA.Among the recognized neurologic diseases in horses, equine protozoal myeloencephalitis (EPM) has been reported around the world and still presents challenges in diagnosis and treatment. Horses can present with clinical neurologic signs consistent with EPM while testing negative for the two main causative agents, Sarcocystis neurona or Neospora hughesi, and may still be clinically responsive to anti-parasitic drug therapy. This context led to our hypothesis that another protozoal parasite, Toxoplasma gondii, which is known to cause toxoplasmosis in other mammalian species, is a potential pathog...
Lew AM, Hosking CS, Studdert MJ.Tests for T- and B-cell quantitation and immune function were developed, and their application in the diagnosis of primary severe combined immunodeficiency disease (CID) in Arabian foals was investigated. Foals with CID had severe lymphopenia and had small or zero numbers of B cells, as shown by immunofluorescence of surface immunoglobulin (Ig), erythrocyte-antibody-complement rosetting, and staphylococcal protein A rosetting. Serum IgM was undetectable in four CID foals 25 to 71 days old. Demonstrable antibody responses were not elicited in CID foals by phage phi X-174, a potent antigen in no...
Fonseca de Araújo Valença SR, Barreto Valença RM, Pinheiro Júnior JW, Feitosa de Albuquerque PP, Souza Neto OL, Mota RA.The aim of the present study was to investigate the occurrence of Toxoplasma gondii among horses and its associated risk factors in Alagoas, Brazil. In total, 440 samples from 36 properties in 23 districts of the state of Alagoas were studied, covering the Leste, Agreste and Sertão mesoregions. Risk factors were evaluated through the application of an investigative questionnaire that focused on the productive, reproductive and sanitary management of herds. T. gondii infection were assayed using the immunofluorescence assay (IFA) with a cutoff point of 64; 14.4% (95% CI: 11.0%-17.8%) of - hors...
Zerpa H, Crawford C, Knight GE, Fordham AF, Janska SE, Peppiatt-Wildman CM, Elliott J, Burnstock G, Wildman SS.The functional distribution of ATP-activated P2 receptors is well characterized for many blood vessels, but not in the equine digital vasculature, which is a superficial vascular bed that displays thermoregulatory functions and has been implicated in ischemia-reperfusion injuries of the hoof. Isolated equine digital arteries (EDA) and veins (EDV) were submitted to isometric tension studies, whereby electric field stimulation (EFS) and concentration-response curves to exogenously applied agonists were constructed under low tone conditions. Additionally, immunofluorescent localization of P2X and...
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.
Böse R, Peymann B, Barbosa IP.Sera from 60 horses held in breeding herd in Brazil were examined monthly by ELISA, immunofluorescence antibody test (IFAT) and Western blot. All foals had maternal antibodies detectable by ELISA and IFAT, and sero-conversion took place between the 2nd and 5th month of age. The 48 and 50 kDa antigens were recognized first in the course of infection. Of 79 sera taken after sero-conversion 78 reacted with the 48 kDa antigen, 76 with the 50 kDa, 50 with the 70 kDa, 54 with the 112 kDa, 72 with the 141 kDa antigen. In general, sera from horses older than 1 year reacted with all 5 diagnostic antige...
Luce R, Shepherd M, Paillot R, Blacklawst B, Wood JL, Kydd JH.An assay has been developed that measures EHV-1 specific interferon gamma synthesis (IFNgamma), a cytokine produced following the activation of memory T lymphocytes and therefore a measure of cell mediated immunity. The method requires validation in the field. Objective: To measure the frequency of EHV-1 specific, IFNgamma synthesising peripheral blood mononuclear cells (PBMC) in a population of Thoroughbred horses, and examine its relationship with age, gender, premises and history of vaccination or field infection with EHV-1. Methods: Lymphocytes from 200 Thoroughbred horses were stimulated ...
Jaspers U, Thiele D, Krauss H.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...
Day MJ, Penhale WJ.Skin biopsies from 47 dogs, 6 cats and 5 horses with suspected autoimmune skin disease were submitted for immunofluorescence from 1978 to 1985. These cases were predominantly Western Australian in origin, although a number were also referred from Queensland and Victoria. In 5 dogs, 2 cats and 2 horses immunoglobulin binding to intercellular cement substance and/or basement membrane was demonstrated by direct immunofluorescence. Antinuclear antibody was also demonstrated in several of these cases. Immunofluorescence was used in combination with histopathological examination to confirm the clini...
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...
Eagle RC, Tortonese DJ.Little is known about the neuroendocrine control of fertility in the horse. In this species, unusual features characterize the normal estrous cycle such as a prolonged preovulatory LH surge during the follicular phase and a distinctive FSH surge during the midluteal phase. This study investigated the distribution and hormonal identity of gonadotrophs in the pars distalis (PD) and pars tuberalis (PT) of the equine pituitary gland as possible morphological bases for the referred unusual endocrine characteristics. In addition, the proportion of gonadotrophs in relation to other pituitary cell typ...
Kamhieh S, Hodgson J, Bode L, Ludwig H, Ward C, Flower RL.Borna disease virus (BDV) is a unique RNA virus that is a cause of neurological disease in horses, sheep and cats. The finding that BDV also infects humans has raised concern related to the impact of infection with this virus. The extent to which BDV may be endemic in geographical regions outside Europe is of interest in management of international movement of animals including horses. Sera from Australian horses (N = 553) sampled in Sydney, New South Wales (NSW), were analysed for BDV antigen, circulating immune complexes (CICs), and antibodies by monoclonal antibody-based ELISAs. One-tenth o...
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...
Jain NC, Vegad JL, Kono CS.Equine neutrophil antibody was raised in rabbits inoculated with equine neutrophils isolated to purity greater than 99.0%, using Percoll density-gradient sedimentation. Neutrophil antibody was detected by use of agar gel diffusion, leukoagglutination, indirect immunofluorescence, staphylococcal protein A and streptococcal protein G binding, and phagocytic inhibition techniques. Precipitin lines and leukoagglutination were seen in antiserum dilutions of 1:4 and 1:64, respectively. The specific nature of leukoagglutination was characterized by the formation of rosette-like clumps of neutrophils....
van den Heuvel LP, van den Born J, Veerkamp JH, Janssen GH, van de Velden TJ, Monnens LA, Schröder CH, Berden JH.1. Proteoglycans extracted from human and equine glomerular basement membranes (GBM) were purified by ion-exchange chromatography and gel filtration. 2. The glycoconjugates had an apparent molecular mass of 200-400 kDa and consisted of 75% protein and 25% glycosaminoglycan. Glycosidase and HNO2 treatment and the amino sugar and sulfate composition of both proteoglycan preparations identified heparan sulfate (HS) as the predominant saccharide chain. 3. Hydrolysis with trifluoromethanesulfonic acid yielded comparable core proteins with molecular masses of ca 160 and 120 kDa. 4. The HS chains had...
Sano Y, Matsuda K, Okamoto M, Takehana K, Hirayama K, Taniyama H.Antigen-presenting cells (APCs) in the uveal tract participate in ocular immunity including immune homeostasis and the pathogenesis of uveitis. In horses, although uveitis is the most common ocular disorder, little is known about ocular immunity, such as the distribution of APCs. In this study, we investigated the distribution of CD163-positive and MHC II-positive cells in the normal equine uveal tract using an immunofluorescence technique. Eleven eyes from 10 Thoroughbred horses aged 1 to 24 years old were used. Indirect immunofluorescence was performed using the primary antibodies CD163, MHC...
Barthold SW, Olson C.Cultured cells from bovine papilloma virus (BPV)-induced fibroblastic tumors and normal dermis of cattle, horses, and hamsters were examined for cell membrane or internal neoantigens, using the indirect immunofluorescence technique. Sera from cattle and horses bearing BPV-induced fibromas cross reacted with cell membranes of tumor, but not with normal dermal cells of both species. The reaction could be blocked with homologous, but not heterologous, serum of these 2 species. Immunofluorescence was not detected with sera from hamsters bearing BPV-induced sarcomas if incubated with bovine, equine...
Hashimoto Y, Ohki H, Sato F, Yanaihara N, Iwanaga T.Chromogranin A (CgA) is an acidic glycoprotein that is co-stored with hormones or neurotransmitters in granular components of endocrine cells and neurons, and released together with them in response to adequate stimulation. In addition to acting as a packaging protein, CgA functions as a precursor molecule that yields several bioactive peptides by proteolytic cleavage. The purpose of this study is to elucidate how different the processing of CgA is among endocrine tissues by immunostaining using multiple region-specific antisera, and to evaluate the availability of region-specific antisera. Wh...
Equine protozoal myeloencephalitis (EPM) is a neurological disease caused by Sarcocystis neurona. Immunofluorescence antibody tests (IFATs) have been widely used to identify exposure of horses to S. neurona in Brazil. Here we used IFAT to search for IgG antibodies against Sarcocystis falcatula-like (Dal-CG23) and S. neurona (SN138) in sera from 342 horses sampled in Campo Grande, Mato Grosso do Sul state (Midwestern), and São Paulo, São Paulo state (Southeastern), Brazil. The 1:25 cutoff value was chosen to maximize sensitivity of the test. IgG antibodies against S. neurona were detected in ...
Green JH, Bolin RC, Carver RK, Gross H, Pigott N, Harrell WK.The serological response in burros and horses to the viable LVS strain of Pasteurella tularensis was studied. High-titered agglutinating antisera and fluorescent-antibody conjugates were obtained in both groups of animals. Maximum titers were obtained in horses 14 to 21 days after the start of vaccination and in burros 21 to 28 days after the start of vaccination. The use of Woodhour's adjuvants or booster inoculations did not result in increased titers.
Minato E, Aoshima K, Kobayashi A, Ohnishi N, Sasaki N, Kimura T.Equine herpesvirus 1 (EHV-1) uses equine major histocompatibility complex class I (MHC class I) as an entry receptor. Exogenous expression of equine MHC class I genes in murine cell lines confers susceptibility to EHV-1 infection. To examine the in vivo role of equine MHC class I as an entry receptor for EHV-1, we generated transgenic (Tg) mice expressing equine MHC class I under the control of the CAG promoter. Equine MHC class I protein was expressed in the liver, spleen, lung, and brain of Tg mice, which was confirmed by Western blot. However, equine MHC class I antigen was only detected in...
