Topic:Complement Fixation
Complement fixation is an immunological reaction involving the binding of complement proteins to antibodies that have attached to specific antigens. In horses, this process is part of the innate immune response, contributing to the identification and elimination of pathogens. The complement system consists of a series of proteins that, when activated, enhance the ability of antibodies and phagocytic cells to clear microbes and damaged cells. This system also promotes inflammation and attacks the pathogen's cell membrane. The complement fixation test is a diagnostic tool used to detect the presence of specific antigens or antibodies in equine serum by observing the consumption of complement proteins. This page compiles peer-reviewed research studies and scholarly articles that explore the mechanisms, regulation, and diagnostic applications of complement fixation in equine immunology.
Conformation of Immunoglobulin M. III. Structural requirements of antigen for complement fixation by equine IgM. Complexes of IgM equine anti-dansyl antibodies and different dansyl substituted carriers were tested for their ability to fix complement (C). Only dansyl92-Ficoll and dansyl12-poly-L-lysine were found to be effective. Dansyl13-bovine serum albumin, dansyl127-keyhole limpet hemocyanin, and reduced and alkylated dansyl10-ribonuclease were all ineffective. Lack of C fixation by the dansyl-ribonuclease was not due to lack of antibody-antigen complex formation, since binding at the concentrations employed for C fixation was established. However, in contrast, polymerized dansyl-ribonuclease (polydis...
Rotavirus infection in foals. Fecal samples from 86 foals with diarrhea were examined by electron microscopy during a 2.5 year period. Of these, 26 (30%) were positive for rotavirus. All of the cases were found in epizootic areas. The disease was produced in an experimental foal by inoculation via stomach tube of a bacteria-free fecal filtrate containing rotavirus. Examination of postmortem tissues from the duodenum and jejunum of 2 naturally infected foals and an experimentally infected foal revealed replicating virus in the intestinal epithelial cells. A limited survey of complement-fixing antibody to rotavirus in horses...
Antibody activities of immunoglobulins in anti-leptospiral horse sera. Antileptospiral sera from hyperimmunized horses were fractionated by gel filtration on Sephadex G-200 or by starch block electrophoresis. The fractions were examined quantitatively for leptospiricidal, agglutinating and complement fixing activities. The leptospiricidal activity was higher in the 78 globulin fraction than in the 19S globulin fraction, while the agglutinating activity was shared by both the fractions being higher in the 19S fraction. Complement fixing activity was found evenly in both the fractions. Leptospiricidal and complement fixing activities were higher in gamma-globulin t...
A comparison of the complement fixation and immunofluorescent antibody tests in a survey of the prevalence of Babesia equi and Babesia caballi in horses in the Sultanate of Oman. 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...
Survey on antibody to Getah virus in horses in Japan. A seroepidemiological survey was performed on antibody against Getah virus in horses in Japan by the complement fixation test. The positive rate was 35 and 53% in two areas where an outbreak of the infectious disease was reported, whereas it was in a range of 3.3 to 24.2% in other areas, except in certain prefectures of the Kyushu district where a high positive rate was observed. In the Hokkaido district, the northernmost part of Japan, no reactors were found in horses under 6 years old, unlike in any other district. It was also suggested that Getah virus infection might have already been prev...
Evaluation of the use of “thermoresistant” antigen Patoc 1, in the diagnosis of human and animal leptospirosis. Preliminary report. The macroagglutination test, according Mailloux, was investigated for its feasibility in the rapid diagnosis of human and animal leptospirosis. Suspected sera examinated by Mailloux test, were also examinated by Complement Fixation and Microagglutination; the results suggest that: Mailloux macroagglutination is the serological test of choice, for screening of animal and human sera, mostly if it is not needed to know the infecting serovar.
Responses of pony mares to the agent of contagious equine metritis 1977. Reproduction of contagious equine metritis 1977 in Pony mares was achieved with cultures of an unclassified Gram-negative coccobacillus. Infected mares developed a vaginal discharge and associated inflammatory changes of the cervix and vagina. There was evidence of variation in pathogenicity between different strains of the organism. Although all infected mares made spontaneous clinical recoveries, the Gram-negative coccobacillus persisted in the genital tracts of a considerable proportion for a variable period after challenge. Recovery of the organism was not associated solely with the occurr...
[Complement fixation reaction studies in rhinopneumonitis of horses]. It was established that the complement binding reaction (CBR) is a suitable and very fast method for horse rhino-pneumonitis diagnostics. Cell cultural virus produced in cell cultures of pig kidneys was used as antigen. The antigen lots tested have no anticomplementary properties. Highest complement binding activity was evident in the non-diluted antigen, which discovered specific antibodies in immune serums. The CBR specificity was tested by the aid of homologous and heterologous serums and antigens. The titers of complement binding antibodies in the serums of 255 horses recovered from the di...
Antibody response of horses to Mycoplasma mycoides subsp capri. In horses given whole cultures or cells of Mycoplasma mycoides subsp capri (by subcutaneous and intravenous injections), antibody responses were measured by serologic procedures. During an immunization period of 22 weeks, horses produced an antiserum that was used to identify M mycoides subsp capri by agglutination, complement-fixation, and fluorescent antibody (FA) tests, but not by the growth-inhibition test. Horses that were injected with whole cultures of M mycoides subsp capri responded better than horses that were injected with only cells, ie, antibodies were detectable sooner by agar ge...
A complement fixation test for antibody to the contagious equine metritis organism. A complement fixation test (CFT) based on that used for brucellosis (Brinley Morgan and others 1971) has been developed for use on the sera of horses exposed to the contagious equine metritis (CEM) organism. None of 50 single samples from horses thought to be unexposed to the CEM organism was positive to the test, although five showed inconclusive reactions. Samples were examined from 41 mares either proved to be infected or from an infected stud. Of these 21 were positive, 11 were inconclusive and nine were negative. The relationship of the CFT to reactions in the other tests used in this con...
The course of serum antibody development in two ponies experimentally infected with contagious metritis. Serum agglutination tests, anti-globulin tests, and complement fixation tests were carried out on sera taken over a period of 98 days from two fillies experimentally infected with the contagious equine metritis organism. The pattern, and significance in diagnosis, of these results is discussed. All 3 tests showed positive titres in the acute phase of experimental disease; reactions in the complement fixation test persisted longest.
Study of homologous and heterologous antibody response in California horses vaccinated with attenuated Venezuelan equine encephalomyelitis vaccine (strain TC-83). Of 359 horses vaccinated with attenuated Venezuelan equine encephalomyelitis (VEE) vaccine (strain TC-83), 87% developed hemagglutination-inhibition (HI) antibodies to VEE virus within 1 month. Blood from a subsample of 101 of the 359 horses was obtained over a 1-year period. Within 1 month after vaccination, 84% of the 101 horses had developed VEE HI antibodies, 87% had developed VEE-neutralizing (Nt) antibodies, and 78% had developed VEE complement-fixing (CF) antibodies. One year after vaccination, 58% of the horses had VEE HI antibodies and 73% had VEE Nt antibodies. The percentage of hors...
Antigenic relatedness of equine herpes virus types 1 and 3. 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...
Immunological properties of two related fragments from human and equine growth hormones. The immunological properties of a synthetic human growth hormone fragment comprising the amino acids 73 through 128 and of the homologous natural horse growth fragment formed by amino acids 73 through 123, have been comparatively studied. Antisera obtained in rabbits inoculated with the native human hormone or with the fragments, were used. By hemagglutination experiments both fragments have the same reactivity toward the anti-human growth hormone serum, but complement fixation curves detect the existence of at least two populations of antibodies presumably originated against the sequence 73-1...
Immunoglobulin G subclass [IgG and IgG(T)] interaction with the P26 group specific antigen of equine infectious anemia virus: immunodiffusion and complement-fixation reactions. Isolated equine immunoglobulin (Ig)G(T) antibodies to equine infectious anemia virus P26 antigen did not precipitate with antigen when the ratio of antibody to antigen was high. However, at lower ratios of antibody to antigen precipitation occurred. In addition, complement-fixation by IgG and P26 antigen was inhibited by high concentrations of IgG(T). The unusual reaction pattern noted with IgG(T) antibodies was still detectable by the immunodiffusion test for equine infectious anemia virus. In situations of nonprecipitability by IgG(T), the adjacent positive control line was inhibited, and th...
[Outbreak of equine influenza by a new strain of Myxovirus type 2. II. Epizootiology]. During an epizootic of equine acute respiratory disease in Algeria, a strain of equine influenza virus was isolated. Sera examination by hemagglutinin inhibition test and complement fixation test confirmed the etiology of the disease. The first and second outbreak of the disease remained localised. The third outbreak spread within few months to all parts of the country. Horses vaccinated with a commercial equine influenza vaccine remained healthy.
Serological relationships between rotaviruses from different species as studied by complement fixation and neutralization. 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...
Serological study of an outbreak of paresis due to equid herpesvirus 1 (EHV-1). Six cases of paresis occurred in a Swedish stud with 48 mares and a stallion. Complement-fixation tests revealed a recent infection with EHV-1 in most horses of the stud. Serumneutralisation tests showed rapid antibody-titre increases during the course of the disease. This type of antibody response was interpreted as induced by reinfection or, possibly, recurrent infection. Two diseased mares were sacrificed. No virus could be isolated from their central nervous system (CNS), liver or spleen, but there is a presumptive evidence for the presence of an antigen specific to EHV-1 in the CNS and li...
[FAB immunoglobulin fragments. I. The comparative characteristics of the serological and virus-neutralizing properties of a gamma globulin against tick-borne encephalitis and of the FAB fragments isolated from it]. A comparative study was made of the serological properties and virus-neutralizing activity of antiencephalitis gamma-globulin and Fab-fragments isolated from it by gel-filtration. Horse immunoglobulins against the autumno-summer tick-borne encephalitis virus could be disintegrated with the aid of papaine to monovalent Fab-fragments which (according to the complement fixation reaction, the test of suppression of the complement fixation, and the HAIT) retained the serological activity whose level was compared with that of the serological activity of gamma-globulin. Fab-fragments possessed a mark...
Detection of immunologically active zones in equine growth hormone. Peptide fragments, obtained from equine growth hormone by cyanogen bromide cleavage and further chemical treatment, were isolated and identified. Their immunological reactivities were tested by hemagglutination and complement fixation methods using rabbit antisera against native hormone. Antigenic determinants were detected in the fragments comprising amino acid sequences 5-72 and 73-123, this last one being predominant. Fragment 124-178 had very low reactivity. Nitration of peptide 73-123 did not modify its immunological properties,but oxidation diminished them. Comparison of the antigenicity...
Serological detection of equid herpesvirus 1 infections of the respiratory tract. 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...
Evidence of respiratory tract infection induced by equine herpesvirus, type 2, in the horse. 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...
Coital exanthema in stallions. Equine coital exanthema can be produced experimentally in stallions by inoculation with an equine herpesvirus (strain 65/61) and be transmitted during coitus with an infected mare. Serological responses to this infection include the production of complement-fixing and serum-neutralizing antibodies which reach maximum levels 14 to 21 days after infection. Complement-fixing antibodies decline rapidly and are usually not detectable by 60 days after infection, whereas serum-neutralizing antibody activity is maintained for at least 1 year. This disparity provides a useful method for the diagnosis o...
Equine infectious anemia: sensitivity of the agar-gel immunodiffusion test, and the direct and the indirect complement-fixation tests for the detection of antibodies in equine serum. The comparative values of the direct, the indirect complement-fixation and the agar-gel immunodiffusion tests were assessed for the diagnosis of equine infectious anemia. Antibodies were detected on the agar-gel immunodiffusion test as early as 18 days post-inoculation in the serums of experimentally infected horses and were readily detectable in all the subsequent bleedings. Complement-fixing antibodies, demonstrable by the direct method, were detected commencing about the same time. However, these were not long-lasting and were replaced by the non-complement-fixing antibodies demonstrable by...