Ten different Salmonella serotypes were grown in a chemically defined medium supplemented with 0.01% yeast extract. After sedimentation of the cells by centrifugation, flagella were detached by exposure to pH 2 for 30 min at room temperature. The flagellaless cells were removed by centrifugation, and the flagellin in the supernatant was further purified by high-speed centrifugation, ammonium sulfate precipitation, and dialysis in 50,000-molecular-weight-cutoff tubing. The 10 flagellin preparations were of a high degree of purity, as demonstrated by electron microscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and measurement of salmonella H and O agglutination titers of antisera raised in rabbits with the flagellin preparations as immunogens.

Actinomyces species are commensal flora usually found in the oropharynx, gastrointestinal tract, and female genital tract. Primary actinomycosis of the breast is an unusual condition, where the most commonly isolated pathogen has been Actinomyces israelii. In recent years, other Actinomyces strains have been found associated with breast disease. We present the first reported cases of breast infection caused by the rare species, Actinomyces turicensis and Actinomyces radingae. Both infections displayed chronicity and abscess formation. In the first case, the infection was refractory to recurrent aspirations and initial antibiotic therapy. In the second case, aspiration and prolonged antibiotic therapy was required to overcome the chronic infection.

Ten antisera raised in rabbits, against polymeric flagellins from ten different Salmonella serotypes were used to determine the relative cross-activities between salmonella flagellins in monomeric form. The results showed a high degree of cross-reactivity between the antisera (IgG antibodies) and all monomeric flagellins investigated. Consequently, it was possible to detect the ten Salmonella serotypes, after heat depolymerization of flagella using radioimmunometric assay with only one antiserum raised against polymeric flagellin from one serotype. The results also showed that native flagella differed antigenically from repolymerized and monomeric flagellins. This is possibly due to changes in the tertiary structure of sub-units of flagella when depolymerized with acid or heat resulting in unfolding and unmasking of common antigenic determinants. The unfolded and unmasked antigenic determinants were not only common to the Salmonella serotypes investigated, but also to other members of Enterobacteriaceae.

Native flagella of ten Salmonella serotypes were shown to possess serotype-specific antigenic determinants as well as a multiple of common antigenic determinants, which varied in concentration. Each common antigenic determinant was shared by certain, but not all, serotypes. This has been demonstrated from agglutination and radioimmunometric assay (RIMA) results, using ten antisera raised in rabbits against purified polymeric flagellins from ten Salmonella serotypes as immunogens. The minimum detectable populations of salmonellae, as determined by RIMA varied considerably, due to variation in concentrations (and possibly types) of common antigenic determinants. However, the results demonstrated the feasibility of utilizing RIMA for the qualitative detection of salmonellae, using a mixture of only the ten antisera and 125I-labelled protein A as a general tracer. In this way, 77 different salmonellae were detected in less than 8 h after culturing in selective broth. The RIMA developed was specific for salmonellae and showed no cross-reactions with high populations of other members within the family Enterobacteriaceae.

Highly purified polymeric flagellin preparations from 10 different Salmonella serotypes were used to produce specific Salmonella H antisera with high titers by the immunization of rabbits. Antigen emulsions in complete Freund adjuvant were administered at the rate of 50 ,ug per rabbit by multiple intradermal injection. Booster injections were given 110 days after the primary immunization. The immune response was monitored regularly over a period of 200 days. The results showed that the H titers, determined with 125I-labeled antigens, averaged 61,000 ± 39,000, and the H agglutination titers of 83% of the animals were >40,000. The high titers of the immunized animals persisted for approximately 4 months. The O agglutination titers of the antisera were <10 for 9 of the fiagellin preparations and ranged from 10 to 320 for the remaining preparation. The antisera obtained were serotype specific after appropriate dilution.