Topic:Antisera
Antisera refer to blood serum containing antibodies against specific antigens, produced by the immune system in response to exposure to these antigens. In horses, antisera are commonly used for therapeutic and diagnostic purposes, particularly in the treatment of venomous bites or stings, and in combating infectious diseases. The production of equine antisera involves immunizing horses with a particular antigen and subsequently collecting and processing their blood to extract the serum rich in antibodies. This page compiles peer-reviewed research studies and scholarly articles that explore the production, application, and efficacy of antisera in equine medicine, as well as advancements in safety and regulatory considerations in their use.
Hemagglutination with equine arteritis virus. Equine arteritis virus (EAV) grown on RK13 cell cultures was tested for hemagglutination (HA) with erythrocytes from a variety of species at 4 degrees C, room temperature and 37 degrees C. HA was observed at all temperatures with erythrocytes from mouse and chicken but not with those of cattle, horse, rabbit, guinea pig, mongolian gerbil, goose or chick embryo. Chickens showed an individual variation in agglutinability of their erythrocytes, requiring selection of birds to obtain erythrocytes for HA. The HA activity was enhanced by treatment of virus materials with Tween 80 followed by treatme...
Production of highly potent horse antivenom against the Thai cobra (Naja kaouthia). Naja kaouthia (NK) causes the highest fatality due to snake venom poisoning in Thailand. The specific antivenom produced is of low potency and in short supply. The aim of this study was to improve the antivenom potency. Bentonite and complete Freund's adjuvants (CFA) and various immunogens were compared. Six groups of three to five horses were immunized as follows: Group 1, NK venom adsorbed on bentonite; Group 2, NK venom in CFA; Group 3, NK venom in CFA in multi-emulsion formulation; Group 4, NK venom in 25% CFA; Group 5, NK neurotoxin 3 (NK3) conjugated with tetanus toxoid (NK3-TT) in CFA; ...
A case of a pony with Cushing’s disease. The pars intermedia of the pituitary gland, and plasma ACTH and cortisol levels in the pony, which was first diagnosed in Japan as indicating equine Cushing's disease, were examined by immunohistochemistry and radioimmunoassay, respectively. The pars intermedia was greatly enlarged and most of its cells were immunoreactive for antisera to both adenocorticotropic hormone (ACTH) and beta-endorphin (beta-End). The plasma ACTH level was elevated when clinical symptoms appeared. The present results reveal that equine Cushing's disease in this pony was induced by the hypersecretion of ACTH and beta-...
Cellular heterogeneity in cathepsin D distribution in equine articular cartilage. The distribution of cathepsin D in normal equine growth cartilage has been examined immunocytochemically using an antiserum raised against human cathepsin D. The cross-reactivity and specificity of the antiserum for equine cathepsin D was confirmed, and its lysosomal localisation was demonstrated in horse skin fibroblasts by confocal scanning microscopy. Cultured horse chondrocytes were heterogenous in their expression of cathepsin D. Heterogeneity of distribution of the enzyme was also seen in chondrocytes in cartilage from different anatomical sites. A high level of cathepsin D was observed ...
Heterologous antisera and antivenins are essential biologicals: perspectives on a worldwide crisis. Active immunization against infectious disease is important. However, much of our world faces poverty, social injustice, and warfare, all of which cause universal immunization to remain a distant dream. Agents that provide passive immunity thus remain essential biologicals. The most important of these are human or equine antisera against rabies, tetanus, diphtheria, and snake antivenins. Homologous products are either unavailable or unaffordable in places where they are needed the most. Less expensive heterologous (equine) antisera can be purified and are safe to use, but these antisera are al...
Ultrastructure of equine morbillivirus. The ultrastructure of the equine morbillivirus (EMV) which was implicated in the death of one human and fourteen horses in Queensland, Australia during September 1994 and a 36 year old man from Queensland in October 1995 is described. The ultrastructure of the virus and the intracellular virus-specific structures are characteristic for the family Paramyxoviridae. Cytoplasmic nucleocapsids were observed within the infected cells monolayers, endothelial cells (lung) of infected horses and the neurons within the brain of the 36 year old Queensland man. Aggregates of smaller nucleocapsid-like stru...
Effect of vaccination of ponies with A4 anti-idiotypic antibody on serum idiotype (1C9) and antilipid A concentration. To evaluate the humoral response of horses to vaccination, using a murine monoclonal anti-idiotypic antibody (A4) that shares an epitope with lipid A. Methods: Serum concentrations of antilipid A antibody and 1C9 (epitope on murine monoclonal antilipid A antibody) were measured serially during the period of vaccination with A4. Methods: 6 clinically normal adult ponies. Methods: Ponies were inoculated IM 3 times at monthly intervals with A4. Two weeks after each inoculation, serum was obtained and was assayed by ELISA for antilipid A and 1C9 concentrations. Additional vaccinations were given t...
Immunohistolocalization of the carbonic anhydrase isoenzymes (CA-I, CA-II, and CA-III) in the reproductive tract of male horses. To elucidate locations of cytosolic carbonic anhydrase isoenzyme (CA-I, CA-II, and CA-III)-positive epithelial cells in equine male reproductive organs. Methods: Descriptive and immunohistochemical study. Methods: 4 clinically normal male horses. Methods: The testis (seminiferous tubules, rete tubules), epididymis (initial, middle, and terminal segments), proximal and distal portions of the ductus deferens, ampulla ductus deferentis, seminal vesicle, prostate, and bulbourethral gland were excised from euthanatized horses after administration of an overdose of pentobarbital. The tissue specimen...
Similarities and differences in supporting and chromaffin cells in the mammalian adrenal medullae: an immunohistochemical study. The adrenal medulla is a typical paraganglion, having the same origin as the sympathetic ganglia, and contains at least two types of parenchymal cells: chromaffin cells and supporting cells. We previously reported that the extent of cellular association of chromaffin cells with supporting cells was remarkably higher in noradrenaline (NA)-than in adrenaline (A)-cell regions in the adrenal medullae of the rat and pig. Methods: Cryostat sections of adrenal medullae of nine mammalian species fixed with Zamboni fluid for 24 h were immunostained by ABC methods using antisera to S-100 protein and PNM...
Antibody-mediated neutralization and binding-reversal studies on alpha-neurotoxins from Micrurus nigrocinctus nigrocinctus (coral snake) venom. An ELISA based, non-radioactive acetylcholine receptor (AchR) binding assay was used to detect the alpha-neurotoxins present in Micrurus nigrocinctus nigrocinctus venom. Sera from horses hyperimmunized against M. nigrocinctus venom contain antibodies which inhibit the binding of M. n. nigrocinctus alpha-neurotoxins to AchR and reverse the binding of toxins already complexed with the receptor. This result supports the importance of using antivenom therapeutically in M. n. nigrocinctus envenomations even after the onset of neurological symptoms. M. nigrocinctus antivenoms cross-reacted in an ELI...
Value of skin testing for predicting reactions to equine rabies immune globulin.
Clinical infectious diseases : an official publication of the Infectious Diseases Society of America
The high cost of postexposure prophylaxis for rabies is one reason that treatment is inadequate in developing countries. This problem has kindled interest in the use of equine rabies immune globulin, which is a less expensive, yet effective, substitute for human rabies immune globulin. Fatal anaphylaxis is a feared complication of the administration of heterologous serum; therefore, authoritative sources recommend prior skin testing. However, recommendations for methods of administering such a skin test and for its interpretation vary greatly. We embarked on a long-term study to develop guidel... Leukocyte alkaline phosphatase and serum gamma-glutamil transferase activities in horses used for production of hyper immune sera. The activities of leukocyte alkaline phosphatase (l-AP) and serum gamma-glutamil transferase (gamma-GT), and total leukocyte counts were determined in horses submitted to the production of hyper immune sera against tetanus (Clostridium tetani). The purpose of this work was to investigate the prospective changes of mentioned parameters in horses under the described circumstances. In addition, the suitability of these parameters in assessing the health condition of the same horses had to be evaluated. The average total leukocyte count increased in one month from the values considered as physiolo...
In vitro and in vivo evidence on the site of neutralization of equine chorionic gonadotrophin (eCG) by an eCG antiserum. This study was designed to determine whether the major site of eCG neutralization by an antiserum to the hormone is at the peripheral or ovarian level. Hamsters hypophysectomized at oestrus were injected s.c. with 25 iu eCG. Three days later, preovulatory follicles were dissected and cultured for 5 h and the medium was changed every hour. At the end of the first hour of incubation, oestradiol and androstenedione accumulation was high, with a sharp drop over the next 4 h, whereas progesterone concentrations did not change over the entire period. Addition of eCG antiserum to the incubated follic...
Immunohistochemical localization of cytochrome P450 aromatase in equine gonads. Estrogens are the major steroids produced by equine gonads. To identify the cells responsible for estrogen synthesis, an antiserum against purified equine testicular cytochrome P450 aromatase was produced in rabbits. The reactivity and specificity of the antiserum were assessed by ELISA, immunoblot analysis, and immunoneutralization studies. Immunofluorescence microscopy demonstrated that in the male gonad, cytochrome P450 aromatase (P450arom) was localized in the interstitial tissue, whereas, under the experimental conditions used, the Sertoli and germ cells did not show any specific staining...
Detection of clenbuterol (Ventipulmin) in the horse. An enzyme linked immunosorbent assay (ELISA) was developed to detect the beta 2-agonist clenbuterol in equine blood and urine. The antiserum was raised in rabbits, employing clenbuterol-diazo-BSA as antigen. Clenbuterol-diazo-horseradish peroxidase served as enzyme conjugate. The concentration of clenbuterol to decrease tracer binding by 50% (IC50 value) was found to be 27.50 +/- 4.20 pg/well (1.37 ng/ml). The antibody cross-reacted with salbutamol (30%), terbutaline (14%) and cimaterol (1%). Horse serum was used directly to screen for clenbuterol, while urine was employed diluted. Positive sc...
Cloning and expression of two genes from Babesia equi merozoites and evaluation of their diagnostic potential. High-titre equine immune sera were used to screen a lambda gt 11 expression library of Babesia equi cDNA fragments. Two cDNA clones which did not cross-hybridize to each other were studied. Both clones hybridized specifically to DNA from B. equi but not to DNA from B. caballi, B. divergens or B. ovis. Recombinant proteins were expressed as glutathione S-transferase (GST) fusion proteins with apparent molecular weights of 40 kDa and 75 kDa. Polyclonal antibodies directed against the 40 kDa and 75 kDa recombinant proteins detected native antigens of 55 kDa and 50 kDa respectively in crude lysate...
Treatment of aplastic anemia with an investigational antilymphocyte serum prepared in rabbits. The authors evaluated antilymphocyte serum prepared in rabbits (ALS-R) as an alternative to antilymphocyte serum prepared in horses (ALG-H) in the therapy of aplastic anemia. Between 1980 and 1993, 57 evaluable patients received ALS-R and prednisone +/- cyclosporine +/- androgens. Standard response criteria were used and patients were evaluated at 3 months from the start of therapy. Median age was 43 years. Disease was present for up to 2 months in 24 patients, 2-5 months in 14 patients, and 6 months or more in 19 patients. Disease was severe in 30 patients and moderate in 27. Responses occurr...
Proteins induced by recombinant equine interferon-beta 1 within equine peripheral blood mononuclear cells and polymorphonuclear neutrophilic granulocytes. Peripheral blood mononuclear cells (PBMC) and polymorphonuclear neutrophilic granulocytes (PMN) as well as embryonic equine dermal fibroblasts and the equine fibroblast line E. Derm which were used as controls, were treated with recombinant equine interferon-beta 1 (rEqIFN-beta 1) in vitro which induced the expression of different proteins in these cells. A 74 kDa protein was induced in PBMC and an 82 kDa protein was additionally found in the equine fibroblast E. Derm cell line following treatment with rEqFN-beta 1. Both proteins reacted with anti-mouse and anti-human Mx protein antisera in im...
Proteolytic processing of the replicase ORF1a protein of equine arteritis virus. To study the proteolytic processing of the equine arteritis virus (EAV) replicase open reading frame 1a (ORF1a) protein, specific antisera were raised in rabbits, with six synthetic peptides and a bacterial fusion protein as antigens. The processing of the EAV ORF1a product in infected cells was analyzed with Western blot (immunoblot) and immunoprecipitation techniques. Additional information was obtained from transient expression of ORF1a cDNA constructs. The 187-kDa ORF1a protein was found to be subject to at least five proteolytic cleavages. The processing scheme, which covers the entire OR...
Effects of polymyxin B and Salmonella typhimurium antiserum on horses given endotoxin intravenously. Polymyxin B and an antiserum against an Re mutant Salmonella typhimurium were evaluated for protective effect in an equine model endotoxemia. Six 3- to 5-month-old foals were given endotoxin (0.25 micrograms/kg of body weight) IV after no pretreatment, or pretreatment with polymyxin B (6,000 U/kg, IV) or S typhimurium antiserum (1.5 ml/kg, IV). When given without pretreatment, endotoxin caused transient recumbency and increases in rectal temperature, and heart and respiratory rates. In addition, leukopenia and increases in circulating tumor necrosis factor (TNF) and interleukin 6 (IL-6) activi...
Expression and characterization of the two outer capsid proteins of African horsesickness virus: the role of VP2 in virus neutralization. African horsesickness virus (AHSV) is a gnat-transmitted member of the Orbivirus genus of the Reoviridae family. The virus has a genome of 10 double-stranded RNA species (L1-L3, M4-M6, S7-S10). The L2 and M6 genes of AHSV serotype 4 (AHSV-4) which encode the outer capsid proteins VP2 and VP5, respectively, were inserted into recombinant baculoviruses downstream of the baculovirus polyhedrin, or p10 promoters. Recombinant baculoviruses expressing VP2, VP5, or VP2 and VP5 proteins of AHSV-4 were isolated. The expressed AHSV proteins were similar in size and antigenic properties to those of viral...
Blood protein polymorphisms in the donkey (Equus asinus). Transferrin, albumin, 6-phosphogluconate dehydrogenase and vitamin D-binding protein polymorphisms were detected in 242 feral and domesticated Australian donkeys by polyacrylamide gel electrophoresis, starch gel electrophoresis, autoradiography, immunoblotting with specific antisera and activity staining. All four TF and two ALB variants were donkey specific while only one of the PGD variants was donkey specific. The two GC variants were electrophoretically identical to the Equus caballus F and S proteins. Available evidence suggested that the TF, ALB, PGD and GC systems are controlled by co-d...
Serological relationship between a donkey alphaherpesvirus (isolate M7/91) and equid herpesvirus type 1 and 4. Rabbit hyperimmune serum prepared against a donkey alphaherpesvirus isolate (M7/91), and against EHV-1 and EHV-4 was used to characterise the antigenic relationship between these 3 viruses. Serum from immunised rabbits was always more specific for homologous virus and showed different cross reactivity for heterologous virus. It was concluded that the immunologic relationship between the M7/91 isolate and EHV-1, was closer than that between this isolate and EHV-4. A serological survey of donkeys (n = 116) and horses (n = 57) revealed evidence of the presence of neutralising antibody to M7/91 in...
Serological and genomic characterization of equine rotavirus VP4 proteins identifies three different P serotypes. A series of viral reassortants was prepared between equine rotaviruses H1 (G5), H2 (G3), and L338 (G13) and human rotavirus ST3 (G4). All contained the VP4 cognate gene segment 4 from the equine parental virus and the VP7 cognate gene segment 9 from ST3. Using these viruses and antisera prepared to them, it was shown that each of the three equine viruses possessed a serologically distinct VP4 or P serotype with a > or = 16-fold difference in reciprocal cross-neutralization titers. H1 VP4 was closely related to that of porcine virus OSU, i.e., P7. L338 gene 4 was sequenced, and the sequence and...
Snake antivenoms from hyperimmunized horses: comparison of the antivenom activity and biological properties of their whole IgG and F(ab’)2 fragments. IgG and F(ab')2 fragments were prepared from horse plasma rich in specific antibodies against Brazilian Bothrops or Crotalus venoms. Both preparations, free of gross contamination with non-immunoglobulin proteins, were able to combine in vitro with their respective antigens, forming immune complexes at antigen excess, equivalence or antibody excess, and activating the C system, through either the classical or the alternative pathways. The IgG preparation was more effective in neutralizing the lethal factors in Bothrops or Crotalus venoms, compared with the F(ab')2 fragments. In contrast, IgG a...
Use of the serum neutralisation test for equine viral arteritis with different virus strains. Serum cross neutralisation tests were conducted with a recent American isolate (84KY-A1) and a European isolate, (Wroclaw-2) and compared with the prototype and modified viruses of the Bucyrus strain of equine arteritis virus by using virus specific immune horse sera. The modified Bucyrus strain was neutralised and showed high neutralisation titres with all the immune sera. The prototype Bucyrus strain was also substantially neutralised, followed by the 84KY-A1 strain. As a result of the tests with the modified Bucyrus strain as the antigen, 20 seropositive horses were discovered among home-br...
A versatile synthetic peptide-based ELISA for identifying antibody epitopes. A simple, versatile and very inexpensive procedure for cross-linking synthetic peptides to the polystyrene surfaces of micro-well assay plates for use in ELISA was developed. The method is based on the use of poly-L-lysine (PLL) as the anchor protein for synthetic peptides which were then easily and covalently linked to the PLL using glutaraldehyde. The synthetic peptides used for the study were based on the amino acid sequence of the equine infectious anemia virus (EIAV) envelope sequence and evaluated as antigens in an ELISA designed to detect antibodies in serum of EIAV-infected horses and ...
Caprylic acid fractionation of hyperimmune horse plasma: description of a simple procedure for antivenom production. A simple methodology for hyperimmune horse plasma fractionation, based on caprylic acid precipitation, is described. Optimal conditions for fractionation were studied; the method gives best results when concentrated caprylic acid was added to plasma, whose pH had been adjusted to 5.8, until a final caprylic acid concentration of 5% was reached. The mixture was vigorously stirred during caprylic acid addition and then for 60 min; afterwards the mixture was filtered. Non-immunoglobulin proteins precipitated in these conditions, whereas a highly enriched immunoglobulin preparation was obtained in...
Identification and verification of the anabolic steroid boldenone in equine blood and urine by HPLC/ELISA. An enzyme linked immunosorbent assay (ELISA) was developed to detect the anabolic steroid boldenone in equine blood and urine. The polyclonal antiserum was raised in rabbits, employing boldenone-17-hemisuccinate-bovine serum albumin as antigen. Boldenone-17-hemisuccinate-horseradish peroxidase served as enzyme conjugate. Sensitivity of the assay was 26.0 +/- 3.0 pg/well. Among the endogenous steroids tested only progesterone and testosterone exhibited moderate cross-reactivities, 3.4 and 2.5%, respectively. These cross-reactivities are of no importance for the boldenone assay. For the reductio...