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Biomedical subjects

J R Etheridge

Publications and source records attributed to J R Etheridge.

At least 19 recordsLinked to original sources

Assessment of a complement fixation test to detect Mycoplasma hyopneumoniae infection in pigs.

Serums from pigs slaughtered at abattoirs were tested for evidence of Mycoplasma hyopneumoniae infection using a complement fixation (CF) test which avoids the procomplementary effect of pig serum. To establish a diagnosis of enzootic pneumonia, the lungs from all sampled pigs were examined for pathological and histological changes consistent with the disease and cultures were made for mycoplasmas and bacteria. The study was carried out at Parkville and Bendigo 160 km apart at different times and all serums were tested at both laboratories. The results agreed closely. Thirty-six of 97 pigs at Parkville and 46 of 99 at Bendigo had enzootic pneumonia. About 80% were positive in the CF test. Sixteen per cent of porkers and 36% of baconers gave false negative reactors, that is, a negative test though lesions were present. About 18% to 36% gave false positive reactions but the level in the porkers in the Bendigo group was significantly higher (p less than 0.02). Possible explanations include, for the false negatives, loss of reactivity caused by circulating antigen and for the false positives, cross reacting antibody produced by another infection or failure to appreciate that lesions of EP were present in lungs because either they were not identified as such or they were not detected. The validity of any serological test for this disease cannot be established while there is a possibility that the present methods used for diagnosis, gross and microscopic examination and recovery of M. hyopneumoniae, fail to detect some infected animals. Other criteria may have to be adopted.

Animals

A method for assessing induced resistance to enzootic pneumonia of pigs.

The resistance induced in pigs by the intravenous, intraperitoneal and subcutaneous inoculation of live Mycoplasma hyopneumoniae was tested by placing inoculated pigs in contact with pigs artificially infected with enzootic pneumonia. Control pigs inoculated by the same routes with sterile culture medium were exposed simultaneously to the infected pigs and the incidence and extent of the pneumonia in both groups were compared. A score derived from the cube root of the calculated or estimated volume of the lung lesion, was used to evaluate severity. Ninety-six per cent of the control pigs developed lesions characteristic of enzootic pneumonia but only 29 per cent of the pigs that had previously been inoculated with M hyopneumoniae developed lesions. Induced lesions differed from those seen in natural cases in extent rather than in character. The method of challenge was satisfactory in that a high proportion of control pigs became infected without the resistance of a majority of inoculated pigs being overwhelmed.

Animals

Production of arthritis by intravenous inoculation of Mycoplasma hyopneumoniae: tests on five strains.

Cultures of five strains of Mycoplasma hyopneumoniae (syn, M suipneumoniae) were each inoculated intravenously into four pigs aged eight to nine weeks. One strain produced no response. For two strains two of the four pigs showed complement fixing antibody and mycoplasmaemia and one of these pigs in each group developed arthritis in one limb joint. The response to the other two strains was more severe: all four pigs in each group had a serological response, were mycoplasmaemic and several limb joints were arthritic.

Animals

A complement-fixation test for enzootic pneumonia of pigs using a complement dilution method.

Complement-fixing antibody to Mycoplasma hyopneumoniae in the serums of pigs experimentally infected with enzootic pneumonia was demonstrated by comparing the haemolytic titre of guinea-pig complement titrated in the presence of heated test serum, M. hyopneumoniae antigen and unheated normal pig serum with the titre obtained when the antigen was omitted. The haemolytic titres against sensitised sheep erythrocytes were determined after a fixation period of 16 to 18 hours at 5 degrees C. When serums, collected at intervals of 3 to 7 days, from 43 pigs exposed to pigs experimentally infected with enzootic pneumonia were tested, 4.6 or more complement units were first fixed 14 to 44 (mean 23.4) days after contact began. Serums collected subsequently fixed from 4.6 to more than 31 complement units. This positive reaction usually persisted until the pigs were killed 4 to 35 weeks after contact began. Thirty-three had gross enzootic pneumonia lesions and 9 had lung lesions detected microscopically. Serum antibody was not detected in 73 weaned pigs aged 7 weeks in a pneumonia-free herd but serums from 9 of 15 unweaned piglets aged 9 to 14 days in the same herd, fixed between 3 and 7 complement units.

Animals

Isolation of Mycoplasma hyopneumoniae from lesions in experimentally infected pigs.

Pigs aged 6 to 9 weeks from enzootic pneumonia-free herds were inoculated intranasally with a suspension of pneumonic lung containing Mycoplasma hyopneumoniae or were placed in contact with such inoculated pigs. All the inoculated pigs had gross lesions of enzootic pneumonia when killed 27 to 42 days after inoculation. The culture methods described enabled M. hyopneumoniae to be isolated from all 29 inoculated pigs. Of 45 pigs in contact with inoculated pigs 35 had gross lesions of enzootic pneumonia when killed 28 to 71 days later and M. hyopneumoniae was isolated from 33. Another 9 had lesions, detected only microscopically, and M. hyopneumoniae was recovered from 3 of these when killed 75 to 98 days after contact began. In a separate experiment M. hyopneumoniae isolated from experimentally infected pigs, and adapted to the culture medium after 6 passages, caused gross lesions of enzootic pneumonia in 1 of 4 pigs inoculated intranasally.

Animals

Studies on the origin of false positive reactions to the complement-fixation test for contagious bovine pleuropneumonia.

Serums of cattle free from contagious bovine pleuropneumonia (CBPP) were tested in complement fixation (CF) tests using 3 antigens; these were the standard antigen (SA) used to test to CBPP in Australia, an ethanol extract antigen (EA) also prepared from Mycoplasma mycoides subsp. mycoides (M. mycoides) and an antigen prepared from a Group 7 bovine mycoplasma isolated from arthritis (AA). The serums included 146 which fixed complement with SA. Eighty percent of these false-positive serums reacted with AA but not with EA; the other 20% were positive at low titre with EA but gave no reaction with AA. Attempts were made to produce false-positive serums experimentally by inoculating 3 Group 7 mycoplasmas and 2 Mycoplasma bovigenitalium strains into cattle. Serums from 3 of 9 cattle inoculated with strain L2917 (Group 7) reacted with SA but differed from the false-positive serums of field cattle by reacting with all 3 antigens. Tests with serums from cattle experimentally infected with CBPP gave similar titres with SA and EA, but the results with AA were mostly negative or less than 10% of the titres obtained with SA and EA. The results of CF tests on serums from the experimental cattle, after absorptions with suspensions of the mycoplasmas, showed that there was a one-way serological relationship between strain L2917 and M. mycoides and between this Group 7 strain and M. bovigenitalium. The CF tests with 3 antigens have assisted in demonstrating the false-positive nature of the reacting field serums encountered in the course of routine CF tests for CBPP in cattle in Australia.

Absorption

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.

Animals