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.
Acland HM, Allen PZ, Kenney RM.After contagious equine metritis bacteria were inoculated into the uterus of mares, genital tract tissues were examined for presence of the organism by bacteriologic cultural technique and an indirect immunofluorescent staining technique. Up to 14 days after mares were inoculated, the organism was frequently in the lumen of the uterus and in the cervix and, less frequently, in the vagina, vestibule, clitoral fossa, clitoral sinus, and uterine tubes. After 21 to 116 days, the organism was occasionally found on the ovarian surface, in the uterine tubes, uterus, cervix, and vagina and more freque...
Klei TR, Chapman MR, Torbert BJ, McClure JR.An indirect fluorescent antibody assay (IFA) was developed using Strongylus vulgaris third stage larvae (L3) as antigens. Observations using the IFA indicate that a species-specific antibody response to S. vulgaris L3 develops in S. vulgaris-infected ponies and that some surface L3 antigens are shared by adult worms. Sequential antibody levels against S. vulgaris were measured in strongyle-naive and in immune ponies following initial and challenge infections using the IFA and an indirect hemagglutination assay (IHA). Antibody levels measured by IFA increased faster following initial infections...
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...
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...
Liu IK, Shivers CA.Cross-reactivity tests indicated that the equine zona pellucida shares common antigen(s) with porcine zona. Porcine oocytes were therefore used to test for the presence of zona antibodies in the sera of horses. Serum samples were collected from 65 mares infertile for unexplained causes: 7 mares were strongly positive for antibodies reactive to porcine zona pellucida as determined by an indirect immunofluorescence method. Of these 7 mares, aged 8-23 years, 2 were nulliparous and 5 were multiparous. Zona-covered horse oocytes were treated with sera from 4 mares previously identified as zona posi...
Wilks CR, Barton MD, Allison JF.Immune responses to Rhodococcus equi were assayed in mares and foals on 7 studs in south-eastern Australia using skin test reactivity to the intradermal injection of culture filtrate and an indirect fluorescent antibody test. The prevalence of positive skin-test reactions did not differ between studs with a history of R. equi disease and those without but there were more mares with high antibody titres on studs with a disease history. A leucocyte extract prepared from mares that were skin-test positive was evaluated for its ability to protect foals exposed to experimental or natural challenge:...
Mitchell G, Liu IK, Perryman LE, Stabenfeldt GH, Hughes JP.Immunoglobulin concentrations were compared in serum and in saline uterine washings from 10 normal mares and 7 subfertile mares with chronic endometrial pathology. Samples were collected in dioestrus and in oestrus. For each immunoglobulin class (IgA, IgG, IgG(T), IgM) the results were expressed as a percentage of total immunoglobulin present in uterine washings or serum. Comparison was also made between changes in immunoglobulin concentrations between dioestrous and oestrous uterine samples, and uterine immunoglobulin concentrations between groups of mares. There was significantly more IgA in...
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...
Monath TP, McLean RG, Cropp CB, Parham GL, Lazuick JS, Calisher CH.Brain tissues were obtained from 5 horses with clinical encephalomyelitis during an epizootic in southwestern Michigan in August-September 1980. These tissues were tested for virus by intracerebral inoculation of suckling mice and by examination of frozen sections and impression smears by the indirect fluorescent antibody (FA) technique. Eastern equine encephalomyelitis virus was isolated and detected by FA technique in brains of 3 horses which died or were euthanatized within approximately 24 hours of onset of the disease but not from 2 horses at 2 and 3 days after onset. The latter 2 animals...
Dutta SK, Bumgardner MK, Scott JC, Myrup AC.Various methods of separation and identification of major equine leukocyte populations and subpopulations were used. The purity of T and B lymphocytes separated in Sephadex anti-equine F(ab')2 columns was 87% to 99% and 83% of 97%, respectively. The purity of T lymphocytes separated in nylon-wool columns was 89% to 98%. Preparations of B lymphocytes separated in glass-bead columns were 68% to 79% pure. The presence (or absence) of surface immunoglobulin by immunofluorescence was the most consistent and reliable method for the identification of B or T lymphocytes, respectively. However, the ery...
Tainturier D, Picavet DP, Badin De Montjoye T, Guaguere J, Tailliar S, Dabernat HJ, Ferney J.Serological response of pony mares to contagious equine metritis is studied comparing three techniques: slow agglutination, complement fixation and indirect immunofluorescence. Sera were taken from pony mares vaccinated with a heat inactivated suspension of Haemophilus equigenitalis, from experimentally-infected pony mares and from healthy horses. All three reactions detected antibodies in vaccinated and infected animals. The highest titers are observed with vaccinated mares. Titers are low in infected animals. Antibodies detected by indirect immunofluorescence appeared sooner and persisted lo...
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...
Williams IF, Heaton A, McCullagh KG.The histological appearance of cells and tissues in the reparative scar tissue which forms in the equine superficial flexor tendon following partial rupture was compared to that of normal tendon. The repair fibroblasts were found to be larger and more basophilic than the tenocytes of normal tendon, to have large vesicular nuclei and to resemble the 'myofibroblasts' described in scar tissue elsewhere. The cell to matrix ratio in scarred zones of tendon was found to be increased and the concentration of collagen in these areas was less than in normal tendon. However, the scar tissue collagen was...
Donnelly J, Joyner LP, Graham-Jones O, Ellis CP.The incidence of antibodies to Babesia equi and B. caballi in horses in the Royal Stables of His Majesty the Sultan of Oman was assessed by complement fixation (CF) and immunofluorescent antibody (IFA) tests. Two series of samples taken with a 2-year interval, mainly from animals reared in Oman, indicated a stable but high prevalence of antibodies. On the 2 occasions 94.6 and 97.7% respectively were positive to B. equi by IFA and 76.8 and 75.0% were positive by CF. For B. caballi the corresponding percentage figures were lower--67.9 and 40.9 by IFA and 30.4 and 40.9 by CF. A group of animals t...
Callow LL, McGregor W, Rodwell BJ, Rogers RJ, Fraser GC, Mahoney DF, Robertson GM.An indirect fluorescent antibody (IFA) test for the diagnosis of Babesia equi infections was evaluated. Antigen prepared by conventional methods was of high quality in one instance and of lesser quality in a second when possible autofluorescence of the horse blood caused inconvenience in reading tests. Tests on 14 horses shown by parasitological means to be either infected (9) or uninfected (5) produced reactions at dilutions of 1/270 to 1/7290 for infected and at 1/10 to 1/90 for uninfected animals. The accuracy of the test was further demonstrated during investigations of 701 horses in 3 sta...
Klevjer-Anderson P, Cheevers WP, Crawford TB.Equine dermal fibroblasts persistently infected with equine infectious anemia virus (EIAV) show no alterations in cell morphology or growth kinetics when compared to uninfected cells. The percentage of cells immunofluorescent positive for viral proteins fluctuated, depending upon the stage of the cell cycle, while production of extracellular virus was uniform throughout the cell cycle, increasing only as the cell number increased. This was shown in log versus stationary phase cultures as well as in cultures synchronized by sterum starvation. The establishment of productive infection did not re...
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...
Dutta SK, Campbell DL.Equine macrophages from the mammary glands of a yearling filly and an 18-year-old barren nonlactatind mare formed cell monolayers in continuous cultures. There was absence of viral cytopathic effect (CPE) in early cell culture passages. The cells from the early cell culture passages having no CPE failed to show evidence of virus or viral antigen by electron microscopic and immunofluorescence studies. Foci of CPE first appeared in the monolayer cell cultures from the filly and the mare in the 3rd and the 4th serial passages respectively, and the CPE increased on subsequent serial passages. Equi...
Woode GN, Bew ME, Dennis MJ.Inoculation at birth with a live attenuated strain of a bovine rotavirus isolated in the USA (scourvax-reo) induced protection in five gnotobiotic calves seven to 21 days later against a UK isolate of pathogenic bovine rotavirus. However, no protection was induced in three calves challenged three to five days after vaccination. There was a close antigenic relationship demonstrated between the two bovine rotavirus isolates. In contrast only one of three gnotobiotic calves inoculated with foal rotavirus, and one of three with human rotavirus, were protected against bovine rotavirus challenge. Pr...
Gutekunst DE, Malmquist WA, Becvar CS.Antiserums prepared in specific pathogen free (SPF) ponies were used in direct and indirect immunofluorescence, immunodiffusion, complement fixation and serum neutralization procedures to study the interrelationships of the three types of equine herpes viruses (EHV-1, EHV-2, and EHV-3). Equine cell cultures infected with each type virus fluoresced when stained with homologous conjugated antiserum. In reciprocal tests EHV-1 and EHV-3 cross-fluoresced, but EHV-2 did not cross-fluoresce. Non-infected cell cultures did not fluoresce when stained with the 3 conjugates. EHV-1 and EHV-3 cross-fluores...
Ito S, Yamada Y, Hayashi M, Matsubara Y.Somatostatin-, glucagon- and insulin-containing cells in the rat and horse pancreatic islets were investigated by an indirect immunofluorescent technique using antibodies to insulin, glucagon and somatostatin. In the rat pancreatic islets, insulin-containing cells were located centrally, and glucagon and somatostatin or somatostatin-like substance (SLS)-containing cells were peripherally disposed and glucagon-containing cells were situated more peripherally as compared with distribution of somatostatin-containing cells. On the other hand, in the horse pancreatic islets, insulin-containing cell...
Ludwig H, Thein P.From 18 horses with clinical symptoms of an affection of the central nervous system and with histopathologic alterations in the brain, four were demonstrated to have Bornavirus-specific antibodies. The antibodies are monospecific, recognizing identical antigens from infected brains of different animal species as well as from persistently infected tissue culture cells. Discrete immunoglobulin species (oligoclonal IgG) can be demonstrated in concentrated horse cerebrospinal fluid; they carry Bornavirus antibody specificity. Their presence, together with the higher antibody titers in the cerebros...
Thouless ME, Bryden AS, Flewett TH, Woode GN, Bridger JC, Snodgrass DR, Herring JA.Human, piglet, mouse, foal, lamb, calf and rabbit rotaviruses all infected, but could not readily be subcultured in LLC MK2 cells. Cells infected with mouse and calf rotaviruses reacted by indirect immunofluorescence (FA) with convalescent serum from children, piglets, mice, foals, lambs, calves or rabbits, taken after rotavirus infection. Human, calf, piglet, mouse and foal rotaviruses reacted with human, calf, mouse, foal and lamb convalescent serum by complement fixation (CF). It was not possible to distinguish between different rotaviruses by CF or FA. Neutralization tests, however, detect...
Thomson GR, Mumford JA, Campbell J, Griffiths L, Clapham P.An investigation was made of 3 serological tests (virus neutralization, complement fixation and indirect immunofluorescence), which are applicable to epidemiological studies of infections by Equid herpesvirus 1 (EHV-1). Sera from gnotobiotic foals inoculated intranasally with various strains of EHV-1 were unable in some cases to neutralize heterologous strains and these results were not consistent with the existence of clearly-defined subtypes of EHV-1, as previously proposed. The cross-reactions in complement-fixation tests paralleled those with neutralization but immunofluorescence tests wer...
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....
Blakeslee JR, Olsen RG, McAllister ES, Fassbender J, Dennis R.Five horses were experimentally exposed to equine herpesvirus 2 strain LK. Two young foals developed chronic pharyngitis (98 and 232 days, respectively). Growth characteristics, cytopathic effects (CPE), inclusion body formation, ether sensitivity, and immunofluorescent analysis indicated that the virus recovered from infected animals was a herpesvirus serologically identical with, or at least antigenically related to EHV-2 strain LK. No significant complement-fixing (CF) or virus-neutralizing (VN) antibody responses were observed in adult horses while both foals demonstrated a rise in CF anti...
Eizema K, van der Wal DE, van den Burg MM, Dingboom EG, Everts ME.An optimal developed musculoskeletal system is vital for the performance of the horse. Previously, we showed that in the m. gluteus medius from adult untrained horses, identical mRNA and protein expression patterns were found in the majority of fibres. However, co-expression of IIa and IId/x myosin heavy chain (MyHC) was substantially more common at the protein than at the mRNA level, suggesting a transcriptionally controlled fine-tuning of these 2 genes. Objective: To analyse the MyHC transcripts and proteins (including the cardiac alpha isoform) in the same muscle during post natal developme...
Newman MJ, Beegle KH, Antczak DF.Monoclonal antibodies to equine lymphocyte antigens were produced, using normal peripheral blood lymphocytes as the immunogen and standard hybridoma techniques. Antibody producing hybridomas were detected by a solid-phase enzyme-linked immunosorbent assay. Antibodies produced by 6 cloned hybrids were characterized further by microlymphocytotoxicity, indirect immunofluorescence, and agglutination assays on peripheral blood lymphocytes, platelets, and erythrocytes. Reaction patterns on leukocytes indicated that these antibodies may recognize at least 3 different cell-surface antigens: (1) an ant...
Hernández-Jáuregui , González-Angulo A.Electron microscopy and immunofluorescence techniques were used to study the various epithelial cells in the endometrial cups of ten pregnant mares, and in the uterine epithelium of one non-pregnant mare. Evidence was obtained which suggested that epithelial gland cells within the endometrial cups, as distinct from the hypertrophied decidual-like cells in the cup, may synthesize PMSG. The suggestion does not agree with the findings of other workers in the fetal origin (chorionic girdle cells) of this gonadotrophin.
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...
el Hasnaoui H, el Harrak M, Tber A, Fikri A, Laghzaoui K, Bikour MH.An indirect fluorescent antibody (IFA) technique was used to screen and quantify antibodies against African horse sickness virus (AHSV) in equine sera. Results obtained with the IFA assay were compared directly with those obtained with standard complement fixation (CF) and virus neutralisation (VN) tests using horse sera from experimental studies and samples from the field. Positive fluorescent antibody titres were detected from as early as 7 days after primary vaccination and persisted for at least six months. The IFA technique offers a clear advantage over CF tests, where the antibodies are ...
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 ...
Tainturier D, Picavet DP, Badin De Montjoye T, Guaguere J, Tailliar S, Dabernat HJ, Ferney J.Serological response of pony mares to contagious equine metritis is studied comparing three techniques: slow agglutination, complement fixation and indirect immunofluorescence. Sera were taken from pony mares vaccinated with a heat inactivated suspension of Haemophilus equigenitalis, from experimentally-infected pony mares and from healthy horses. All three reactions detected antibodies in vaccinated and infected animals. The highest titers are observed with vaccinated mares. Titers are low in infected animals. Antibodies detected by indirect immunofluorescence appeared sooner and persisted lo...
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...
Iqbal J, Edington N.Equid herpesvirus 1 (EHV-1) is the most common cause of virus-induced abortion in horses. After primary infection the virus becomes latent predominantly in the respiratory tract lymph nodes and the genome can also be detected in the peripheral nervous system. The role of mouse as a feasible model for the establishment of latency and reactivation of EHV-1 was investigated. Intracerebral and intranasal infections of 3- and 17-day-old mice were made and virus replication was confirmed by virus isolation and detected by indirect immunofluorescence (IIF) in brain. For reactivation studies, the mice...
Metz GE, Serena MS, Ocampos GM, Panei CJ, Fernandez VL, Echeverría MG.Equine arteritis virus (EAV) was isolated from a testicle of the presumable first stallion infected with EAV in Argentina. This virus isolate (named LT-LP-ARG) was confirmed by GP5-specific PCR and indirect immunofluorescence assays. The PCR product was sequenced, and the phylogenetic analysis revealed that the LT-LP-ARG strain of EAV forms a monophyletic group, together with other strains previously isolated in our laboratory (LP02 group). However, all Argentinean EAV strains belong to a polyphyletic group. We believe that the virus isolate presented in this report could be the origin of EAV ...
Costa PWL, Oliveira CSM, Bezerra RA, Alvares FBV, Formiga VHAS, Martins MRDD, Feitosa TF, Vilela VLR.The aim of this study was to describe the prevalence of anti- and anti- antibodies in equids that carry out traction work in Northeastern Brazil, and to establish the potential risk factors associated with seropositivity for these agents. Blood samples were collected from 322 traction equids (horses, donkeys and mules) in urban areas of 16 municipalities in the State of Paraíba, Brazil. The samples were sent for serological diagnosis using the Immunofluorescence Antibody Test (IFAT). Epidemiological questionnaires were given to the owners to assess the possible risk factors associated with in...
Scott DW, Walton DK, Smith CA, Lewis RM.Indirect immunofluorescence testing for pemphigus-like antibodies was performed on 79 horses: 28 horses with various nonpemphigus dermatologic diseases, 21 horses with various nondermatologic diseases, and 30 normal horses. Pemphigus-like antibodies were detected in 6 horses: 3 normal horses with titers of 1:40, 2 horses with dermatophilosis at titers of 1:10 and 1:80, and 1 horse with lymphosarcoma at a titer of 1:320. It was concluded that equine pemphigus-like antibodies are a potential source of misinterpretation and misdiagnosis in indirect immunofluorescence testing. Direct immunofluores...
Atwill ER, Mohammed HO, Lopez JW.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...
Do Carmo GM, Da Silva AS, Klauck V, Pazinato R, Moura AB, Duarte T, Duarte MM, Bochi GV, Moresco RN, Stefani LM.Toxoplasmosis is an important parasitic disease affecting several species of mammals, but little is known about this disease in horses. This study aimed to investigate the levels of several immunological variables and markers of cell damage in the serum of seropositive horses for Toxoplasma gondii. Sera samples of adult horses from the Santa Catarina State, Brazil used on a previous study were divided into groups according to their antibody levels for T. gondii determined by immunofluorescence assay, i.e. 20 samples from seronegative horses (Group A - control), 20 samples from horses with tite...
Vĕzník Z, Svecová D, Pospísil L, Diblíková I.Frequency of elementary and reticular chlamydial bodies was investigated by direct immunofluorescence tests in ejaculates collected from 52 men, 60 stallions, 42 bulls, and 66 boars using the kits of Progen Biotechnic GmbH and the microscope Labophot-2 Nikon. At the same time, qualitative semen tests, including ejaculate volume, sperm motility, percentage of live and dead sperms and morphological' analyses (Vĕzník and Svecová, 1992) were done. Repeatability of the findings was checked in a group of nine bulls housed at the institute and sampled at weekly intervals for 3 to 4 months (Tab. 1)...
Yeşilbağ K, Herzog S, Kennerman E, Tuncer P, Schmid S, Kaya G, Thiel HJ.Distribution of Borna disease virus (BDV) infection outside endemic areas has been studied in several countries. We examined serum samples for anti-BDV antibodies in purebred racing horses and other domestic animals in Turkey. In total serum samples of 437 animals including 282 horses, 50 sheep, 25 goats, 50 cattle, and 30 cats were tested by indirect immunofluorescence assay (IFA). Anti-BDV antibodies were detected in 4.9% of horses, 12% of sheep, 4% of goats, 14% of cattle and 6.6% of cats. No statistical difference was observed between seroprevalence in Arabic and English purebred horses fr...
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.
Sim RR, Padilla LR, Joyner PH, Anikis P, Aitken-Palmer C.Anaplasma phagocytophilum (formerly Ehrlichia equi ) is a tickborne pathogen of domestic horses and the causative agent of equine granulocytic anaplasmosis. After the occurrence of clinical anaplasmosis in a Przewalski's horse ( Equus ferus przewalskii) housed at the Smithsonian Conservation Biology Institute in 2008, opportunistic serosurveillance of the herd was initiated. From 2008 to 2014, 57 serum samples were collected from 27 individuals (10 males; 17 females). Using indirect immunofluorescent antibody assays for anti- Anaplasma phagocytophilum antibodies, it was determined that prevale...
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...
Liu X, Liu Q, Feng X, Tang Q, Wang Z, Li S, Feng Z, Zhu J, Guan X.Due to the disadvantages of human and equine rabies immunoglobulin, it is necessary to develop a substitute for HRIG and ERIG, especially for those people living in the developing countries. Because of higher affinity and lower immunogenicity of rabbit's immunoglobulins, anti-rabies immunoglobulins specific to rabies virus were produced in rabbits as a bioreactor, and had been characterized by ELISA, affinity assay, immunofluorescence assay (IFA), immunocytochemistry, rapid fluorescent focus inhibition test (RFFIT). ELISA, affinity assay and IFA showed that rabbit RIG (RRIG) bound specifically...
Otto S, Michler JK, Dhein S, Mülling CKW.Distal axonopathy is seen in a broad range of species including equine patients. In horses, this degenerative disorder of the recurrent laryngeal nerve is described as recurrent laryngeal neuropathy (RLN). The dysfunctional innervation of the cricoarytenoideus dorsalis muscle (CAD) leads to a loss of performance in affected horses. In general, ex vivo models of the larynx are rare and for equine patients, just one short report is available. To allow for testing new therapy approaches in an isolated organ model, we examined equine larynges in a constant pressure perfused setup. In order to chec...
Meyers SA, Rosenberger A, Orpneck K.Three protein bands with hyaluronidase activity and molecular masses of 87, 48 and 43 kDa were isolated from purified equine sperm plasma membranes. Indirect immunofluorescence was used to assess sperm labelling patterns using a polyclonal antibody to sperm hyaluronidase. In ejaculated spermatozoa, surface-associated hyaluronidase was localized to the posterior head region of 98 +/- 2% of spermatozoa (n=10). Epididymides were isolated from mature stallions (n=5) and divided into caput, corpus and cauda epididymides in separate Petri dishes. The epididymidal tubules were dissected and washed us...
Bombardi C, Grandis A, Gardini A, Sorteni C, Clavenzani P, Chiocchetti R.The activity of the gastrointestinal tract is regulated through the activation of adrenergic receptors (ARs). Since data concerning the distribution of ARs in the horse intestine is virtually absent, we investigated the distribution of β2-AR in the horse ileum using double-immunofluorescence. The β2-AR-immunoreactivity (IR) was observed in most (95%) neurons located in submucosal plexus (SMP) and in few (8%) neurons of the myenteric plexus (MP). Tyrosine hydroxylase (TH)-IR fibers were observed close to neurons expressing β2-AR-IR. Since β2-AR is virtually expressed in most neurons located...
Fraser NS, Wilborn RR, Johnson AK, Braden TD, Wiley AA, Canisso IF, Bartol FF.Equine uterine development, including endometrial histogenesis, begins prenatally and is completed postnatally. Little is known about this process in the horse. Uterine tissue was acquired from 38 foals, ranging in developmental age from gestational day (GD) 300 to postnatal day (PND) 180, for assessment of endometrial histogenesis. Patterns of endometrial cell proliferation were evaluated by multispectral imaging of uterine tissue sections stained immunofluorescently for Ki-67. Labeling index (LI, % labeled cells) for Ki-67 was calculated for each endometrial cell compartment (luminal epithel...
Guimarães AM, Bruhn FRP, Ribeiro MJM, Rosa MHF, de Mello Garcia A, da Rocha CMBM, de Assis Rocha I.Theileria equi, a protozoon in the phylum Apicomplexa, is the causative agent of equine theileriosis. The aim of this cross-sectional study was to determine the prevalence of IgG antibodies against T. equi, by using the indirect fluorescent antibody test (IFAT) reaction and correlating the serostatus with some epidemiological variables relating to the way in which Mangalarga Marchador horses are raised in southern Minas Gerais, Brazil. In this study, 506 horses were used, all clinically healthy, on 53 horse farms distributed across 27 municipalities in southern Minas Gerais. The statistical te...
Heath E, Aire T, Fujiwara K.A microtubular mass (MM) defect was found in the spermatozoa of 7 Standardbred stallions; 3 stallions were sons of the same sire. Two of these 3 stallions and 2 other stallions (for a total of 4 out of the 7 stallions) were considered subfertile when the defect was first observed. Fertility improved with time, either during the first breeding season or when a given stallion was used less frequently; however, the MM defect persisted, consisting of tortuous arrays of small abnormal microtubules visible only by transmission electron microscopy. The MM probably contained the protein tubulin as ind...
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...
Breuil MF, Duquesne F, Sévin C, Laugier C, Petry S.Contagious equine metritis is a horse disease that causes endometrial inflammation due to Taylorella equigenitalis. Since Taylorella asinigenitalis was characterized, genital swab culture has proved to be an insufficient method for distinguishing between the two Taylorella species. Here, we developed an indirect immunofluorescence (IIF) test using polyclonal antibodies. Specificity, sensitivity, and detection limit were assessed using isolated bacteria (55 T. equigenitalis strains, 46 T. asinigenitalis strains and 18 other bacterial species), experimental and genital swabs in comparison to bac...
Veerasammy B, Gonzalez G, Báez-Ramos P, Schaaf CR, Stewart AS, Ludwig EK, McKinney-Aguirre C, Freund J, Robertson J, Gonzalez LM.Intestinal epithelial stem cells (ISC) are responsible for epithelial regeneration and are critical to the intestine's ability to regain barrier function following injury. Evaluating ISC biomarker expression in cases of small intestinal strangulation (SIS) may provide insight into clinical progression. Objective: Intestinal resection margins from cases of SIS were evaluated to determine if (1) evidence of injury could be identified using histomorphometry, (2) ISC biomarker expression was decreased in the proximal resection margin compared to control and distal resection margin, and (3) the ISC...
Wada R, Fukunaga Y, Kanemaru T, Kondo T.Five pregnant mares, at between 6 and 8 months gestation, were experimentally infected with the Bucyrus strain of equine arteritis virus (EAV). Of the five mares, four aborted and one died. The pathogenesis of the abortions was studied, using histopathologic techniques, tissue immunofluorescence and virus isolation. Common microscopic lesions in the maternal reproductive organs indicated myometritis with a degeneration of the myocytes and an infiltration of the mononuclear cells. Epithelial cells of the endometrial gland showed sporadic degeneration. Lesions in the fetal tissue included an atr...
