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Evaluation of a selective medium for isolation of Haemophilus pleuropneumoniae


, : Evaluation of a selective medium for isolation of Haemophilus pleuropneumoniae. Canadian Journal of Comparative Medicine 47(4): 445-450

Crystal violet, lincomycin, spectinomycin and bacitracin were evaluated as selective agents in media for isolation of H. pleuropneumoniae. No single antimicrobial agent or combination of 2 or more inhibited all non-Haemophilus strains (Escherichia coli, Pasteurella haemolytica, P. multocida, Streptococcus faecalis, S. equisimilis and Staphylococcus aureus) without marked suppression of 16 H. pleuropneumoniae strains. A medium containing 1 .mu.g/ml of crystal violet, 1 .mu.g/ml of lincomycin, 8 .mu.g/ml of spectinomycin and 128 .mu.g/ml of bacitracin inhibited 1 E. coli strain and the gram-positive strains while H. pleuropneumoniae strains were suppressed to a minor degree only. H. pleuropneumoniae was isolated on the selective medium on 3 occasions from the nose or pharynx of 2 of 8 experimentally inoculated pigs. H. pleuropneumoniae was recovered from the nose of only 2 pigs at necropsy and from tonsil of 1; the lower airways in most pigs and the lung lesions in all pigs were positive. There was no advantage to using the selective medium for the recovery of H. pleuropneumoniae at necroscopy from these 8 experimentally infected pigs, probably because other bacteria were absent or present in very low numbers in the tissues with H. pleuropneumoniae. The isolation rate on selective medium was higher than the rate on non-selective medium (P .ltoreq. 0.1; .chi.2 test) when the airways of slaughtered pigs were cultured. This was likely due to a high degree of contamination. Dry swabs placed in tryptone yeast extract with nicotinamide-adenine-dinucleotide gave a significantly higher recovery rate than commercial Culturette swabs in modified Stuart's transport medium. This difference was more marked after 24 h storage at room temperature, suggesting that the Culturette swab is not suitable for transporting samples to diagnostic laboratories for H. pleuropneumoniae culture. Fifty-one Haemophilus strains were isolated from 224 slaughtered pigs and identified as H. pleuropneumoniae serotype 1 (5 strains), serotype 5 (9 strains), serotype 7 (18 strains), minor group identical with strain 202 (4 strains) and nontypeable (15 strains).

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Accession: 005395674

PMID: 6230143

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