Acute respiratory, circulatory and pathological changes in the calf after intravenous injections of the galactan from Mycoplasma mycoides subsp. mycoides.

Twenty of 28 calves, 10-12 weeks of age when given intravenous injections of the galactan from Mycoplasma mycoides subsp. mycoides, showed transient apnoea, increased pulmonary arterial and decreased systemic arterial blood pressures, and increased packed-cell volume. Necropsy revealed haemorrhages associated with alveolar ducts and vessel walls, areas of pulmonary oedema, usually associated with the haemorrhages, dilated airways and, in some, capillary thrombosis. Animals that had shown changes in blood pressure and respiration in response to a dose of galactan did not react to a second dose an hour later. One goat tested died, four lambs were mildly affected and a cat and several rats and guinea-pigs did not respond. It is suggested that the galactan released biogenic amines that produced the effects listed. Immunological mechanisms were discounted on the grounds that only a small amount of antigenic material was injected at the time the reaction occurred, and neither serological nor skin tests produced any evidence of prior sensitisation to the galactan or a similar substance. A relationship between reactivity to the galactan and susceptibility to the natural disease has been suggested. This, together with the pulmonary oedema found in galactan-treated calves and in natural lesions of contagious bovine pleuropneumonia (CBPP), and the possibility that contraction of blood vessels could be an initiating cause of thrombosis indicates the role that galactan may play in the pathogenesis of CBPP.

Improving the specificity and yield of the contagious bovine pleuropneumonia complement fixation test antigen.

Several methods for increasing yield and specificity of the contagious bovine pleuropneumonia complement fixation test antigen which is derived from Mycoplasma mycoides subsp mycoides, strain V5, were examined. Changes in culture conditions that increased the cell mass per unit volume of culture did not result in comparable increase in antigen yield. Sixteen to 60-day-old cultures yielded more boiled cell antigen than younger cultures. Some antigen in young cultures appeared to be masked, probably by galactan. The yield of antigen extracted from boiled cells with ethonol was as much for two to eight-day-old cultures as for older cultures. The ethanol extract antigen was less reactive with false positive bovine sera than standard boiled antigen while reactivity with anti Mycoplasma mycoides sera was similar to that of standard antigen. Adsorbed gamma globulin was not detected in either boiled or ethanol extract antigen. The data suggest that several complement fixing antigens were present in antigens derived from older cultures.

Reverse phase passive haemagglutination and single radial immunodiffusion to detect epsilon antigen of Clostridium perfringens type D.

Two in vitro immunological assays were developed for detection of the epsilon (epsilon) antigen of Cl. perfringens type D. It was found that the reverse phase passive haemagglutination assay (RPHA) was able to detect concentrations of epsilon-antigen as low as 6 x 10-7 mg/ml whereas the single radial immunodiffusion techniques (SRID) was capable of detecting concentrations of epsilon-antigen above 0.01 mg/ml. When applied to gut contents from freshly dead infected sheep the RPHA test was found to be more sensitive than mouse toxicity assay in detecting the presence of epsilon-antigen. However, very low titres were detected in gut contents from normal sheep which meant that in a diagnostic situation interpretation of RPHA titres would be difficult. No epsilon-antigen was detected by SRID in gut contents from normal sheep or in gut contents from freshly dead infected sheep. The SRID assay could detect epsilon-antigen in gut contents from infected sheep allowed to decompose for 20 h post-mortem.

Human immunoglobulin subclasses. Partial amino acid sequence of the constant region of a gamma 4 chain.

The heavy chain of a human myeloma protein (Vin) belonging to the gamma4 subclass was subjected to tryptic digestion after reduction and carboxymethylation. Cyanogen bromide fragments were also prepared and all 19 tryptic peptides that account for one of them (the Fc-like fragment) were studied. Selected peptic peptides were isolated and provided evidence for the order of 15 of the tryptic peptides. In addition the sequence of two large peptic peptides derived from two sections of the molecule including all the interchain bridges is presented. Comparison with published data on other chains allows us to propose a sequence of gamma4 chains that extends from just before the presumed starting point of the invariable region (at about residue 113) to the C-terminal end of the chain (approx. residue 446), except for a section of about 50 residues. The results of the comparison suggest that the immunoglobulin subclasses have a recent independent evolutionary origin in different species. Implications for complement fixation and for the evolutionary origin of antibody diversity are also discussed.