A solid-phase immunoassay for detection of IgA1 protease activity on agar plates.

An immunoassay was developed which is capable of detecting IgA1 protease positive microorganisms from biologically derived samples grown on agar plates. Activity was detected by overlaying plates with agar containing insolubilized IgA and capturing Fab fragments on nitrocellulose disks coated with anti-lambda light chain. Using cultures of human saliva, a comparison of this assay with a more traditional method involving electrophoresis and Western blotting of individual colony material revealed excellent agreement between the two methods. This assay should be useful in studying the distribution and ecology of organisms producing IgA1 protease, and in assessing its potential as a virulence factor.

Antimicrobial resistance of fecal coliforms from pigs in a herd not exposed to antimicrobial agents for 126 months.

Patterns of antimicrobial resistance were determined for lactose-fermenting fecal coliforms obtained during a 20-month period from pigs in a herd without exposure to antimicrobial drugs for 126 months. Mean percent resistance to cephalothin, chloramphenicol, kanamycin, naladixic acid, streptomycin and tetracycline was lower (P less than 0.01) for isolates obtained during the 20-month period (Group 2) than for isolates obtained during the first 13 months (Group 1) after the withdrawal of antibiotics. Mean tetracycline resistance was 40.5% for Group 2 isolates. Approximately 44% of the Group 2 isolates were resistant to the 10 antimicrobial agents compared with 74.9% for the Group 1 isolates (P less than 0.01). Multiple resistance was lower (P less than 0.01) in the Group 2 isolates than in the Group 1 isolates. Four resistance patterns (tetracycline, tetracycline-streptomycin, tetracycline-sulfisoxazole, tetracycline-streptomycin-sulfisoxazole) accounted for 74% of the resistant isolates.

Effect of age and housing location on antibiotic resistance of fecal coliforms from pigs in a non-antibiotic-exposed herd.

The relationship of age and housing location to single antibiotic resistance, multiple antibiotic resistance, and resistance patterns of fecal coliforms obtained during a 20-month period from pigs in a herd that was not exposed to antibiotics for 126 months was determined. Bacteria resistant to single and multiple antibiotics were isolated more frequently (P less than 0.01) from pigs under 7 months of age. A greater proportion of isolates from pigs over 6 months of age was sensitive to the 13 antimicrobial agents tested (P less than 0.01), while a smaller proportion showed resistance to single (P less than 0.05) and multiple (P less than 0.01) antibiotics. More than 80% of the resistant isolates were resistant to tetracycline, streptomycin, or sulfisoxazole. Resistance was greater (P less than 0.01) for pigs in the finishing unit than for those on pasture. Resistance to ampicillin, carbenicillin, and tetracycline was greater (P less than 0.05) for pigs in the finishing unit than for those in the farrowing house. More isolates from pigs on pasture were sensitive to all antimicrobial agents tested (P less than 0.01). A greater proportion of isolates from pigs in the finishing unit showed resistance to a single antibiotic (P less than 0.01). The data from this study suggest that exposure to antibiotics is not the only factor that influences the prevalence of bacteria that are resistant to single and multiple antibiotics in the feces of domestic animals and that considerable research is needed to define the factors influencing antibiotic resistance in fecal bacteria.