Dilution technique for isolation of Haemophilus from swine lungs collected at slaughter

A total of 307 lungs obtained from a slaughterhouse were cultured by a dilution technique for the isolation of Haemophilus spp. The technique consisted of performing serial (10-fold) dilutions of the tissue samples to a dilution of 10(-5). Two selective media were used. L broth consisted of a basal brain heart infusion broth containing 5% horse serum, 5% yeast extract, and 100 micrograms of NAD and 0.5 microgram of lincomycin per ml. L-B broth was identical to L broth, except 1.5 microgram of bacitracin per ml was included. The broths were incubated overnight and then plated onto blood agar. A total of 83 (27%) isolates were obtained, and both media proved to be necessary, as a proportion of isolates grew in one of the media employed but not in the other. Of the isolates, 66.3% were urease positive and most of these (98%) were classified as "minor group" strains. Urease-negative strains (27.7%) were classified as Haemophilus parasuis.