Brucella abortus strains were isolated from bovine tissue and milk samples from seven Ontario herds. The isolates were characterized by colonial morphology, requirement of CO2 for growth, lysis by Tbilisi phage, biochemical tests and agglutination in monospecific sera. They resembled B. abortus biotype 2 (on the basis of sensitivity to thionin and basic fuchsin) and biotype 4 (on the basis of agglutination with anti-Brucella "M" but not anti-Brucella "A" absorbed sera). Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of these isolates and B. abortus biotypes 1, 2 and 4 showed similar profiles. Immunoblots with anti-A and anti-M absorbed sera showed different antigenic regions reacting with the specific sera and also confirmed that the atypical B. abortus isolates were serologically similar to biotype 4.
An hemolysis-in-gel test (HIGT) for bovine antibody against Brucella abortus was developed and evaluated. Sera to be tested were placed in wells in an agarose gel containing guinea pig complement and J-negative bovine erythrocytes coated with lipopolysaccharide prepared from B. abortus biotype 1. After incubation, zones of hemolysis were produced by positive sera. The activity of some positive sera was heat labile, but was restored to heated sera by addition of a crude preparation of the first component of bovine complement. The HIGT was compared with the complement fixation test (CFT), the buffered antigen plate test (BPAT), the standard tube agglutination test (STAT) and the standard plate agglutination test (SPAT), using sera from 1041 brucellosis-free cattle, from 51 cattle infected with B. abortus biotype 1 or biotype 4, and from six heifers vaccinated with strain 19. Three of 1041 sera (0.29%) from brucellosis-free cattle were HIGT-positive, ten (0.96%) were BPAT-positive, and none were positive in the CFT, STAT, or SPAT. The HIGT was more sensitive, and detected infection earlier than the other tests in the case of B. abortus biotype 1 infection, but was less sensitive for biotype 4 infection. In vaccinated heifers, HIGT reactions appeared later than reactions to the other tests, and persisted as long as 565 days. Further studies are needed to standardize the HIGT test conditions and to improve its sensitivity for B. abortus biotype 4 infections. Attempts to determine the effects of LPS antigen prepared from different biotypes of B. abortus are in progress.
Three flocks raised for broiler or roaster performance tests were studied to determine the incidence and sources of salmonellae during the growing period, transport and processing and to relate these to contamination of processed carcasses. Day old chicks in two of the tests, (tests IV and V), were treated with a culture of intestinal anaerobes derived from mature chickens. The incidence of salmonellae during the growing period was too low to permit any conclusions about the efficacy of this culture in preventing Salmonella infection, but it had no adverse effect on flock performance. Carcasses from all three flocks were contaminated with salmonellae. Although the test IV flock was raised free of salmonellae, 46% of the carcasses tested from this flock were contaminated. The apparent source was the transport crates, 99% of which yielded salmonellae before the flock was loaded. In test V, 92% of the carcasses tested yielded salmonellae. The apparent sources were: flock infection (apparently originating from the parent flock), contaminated crates, spread during transport, and plant contamination. The flock of test VI was infected with Salmonella albany, and 54% of the carcasses tested were contaminated with this serovar. Carcasses of chicks infected early in life were more likely to be contaminated than those of chickens which contacted salmonellae later in the growing period.
Rinse fluids collected from eight different lots of commercially processed broiler carcasses were cultured for salmonellae by a meat probable number method. Most probable number values of 80 or less were obtained from all carcasses in three of the eight lots, but two lots contained carcasses which yielded more than 1000 salmonellae. Similar numbers of salmonellae were recovered in repeated rinses of the same carcasses, suggesting that only a small proportion of the total population was recovered in each rinse. Five lots originated from flocks of known Salmonella status. More salmonella were recovered from carcasses which originated from flocks infected in the first week of life than from flocks infected at a later date. Carcasses from a flock which was processed immediately after a contaminated flock yielded more salmonellae than carcasses processed at the beginning of the day in a clean plant.