Confirmation that PCR can be used to identify NAD-dependent and NAD-independent Haemophilus paragallinarum isolates.

Seventy five bacteria tentatively identified as Haemophilus paragallinarum (the causative agent of infectious coryza), eight identified as Ornithobacterium rhinotracheale and 13 identified as NAD-independent Pasteurella species were isolated from chickens with respiratory infection in various provinces in South Africa. The isolates were characterized by conventional biochemical and serological methods. A polymerase chain reaction (PCR) assay specific for H. paragallinarum was used to identify the cultures directly from colonies. The PCR assay gave positive results for all isolates that were identified by conventional methods as H. paragallinarum, irrespective of whether they were nicotinamide adenine dinucleotide (NAD)-dependent (43 isolates) or NAD-independent (32 isolates). The eight isolates that were identified by conventional methods as O. rhinotracheale and the 13 isolates identified as various Pasteurella species gave negative results in the PCR assay. This study has demonstrated that colony PCR is a rapid method for uniquely identifying both NAD-dependent and NAD-independent strains of H. paragallinarum and distinguishing them from other bacteria, such as O. rhinotracheale and Pasteurella species.

Correlation between ability of Ornithobacterium rhinotracheale to agglutinate red blood cells and susceptibility to fosfomycin.

Twenty five freeze-dried isolates of Ornithobacterium rhinotracheale were used for the determination of minimum inhibitory concentrations (MIC) against the antibiotic fosfomycin (Fosbac, produced by Bedson SA, consisting of a 25% mixture of fosfomycin). The same isolates were tested for their ability to haemagglutinate chicken red blood cells. Ten of the 25 isolates were found to be susceptible to fosfomycin (MIC values below 128 ug/ml). All of these isolates were able to agglutinate red blood cells. This is the first report on the ability of O. rhinotracheale to agglutinate red blood cells. The remaining 15 isolates were resistant to fosfomycin (MIC values above 128 ug/ml). Only five of these isolates were found to have the ability to agglutinate red blood cells. There appears to be a correlation between the ability of O. rhinotracheale isolates to agglutinate red blood cells and their susceptibility to fosfomycin. The ability of certain isolates of O. rhinotracheale to agglutinate red blood cells, raises the questions of differences in virulence between the isolates which can agglutinate red blood cells and those which cannot and the use of this ability to agglutinate red blood cells as an alternative method for serotyping O. rhinotracheale.

Isolation and identification of NAD-independent bacteria from chickens with symptoms of infectious coryza.

Since 1990, NAD-independent bacteria have been isolated in South Africa from poultry showing respiratory manifestations similar to infectious coryza. A total of 126 isolates was examined biochemically and serologically, using polyclonal as well as monoclonal antibodies. Forty isolates were identified as Ornithobacterium rhinotra-cheale, some of which agglutinated glutaraldehyde-fixed red blood cells. Furthermore, fourteen Pasteurella avium isolates, five P. volantium and three Pasteurella species A were isolated for the first time. The remaining 64 isolates were biochemically identified as NAD-independent H. paragallinarum. Of these, 37 were Page serovar A, while no Page serovar B isolates were found. The remaining 25 isolates were typed as Page serovar C. Two different haemagglutination inhibition reaction patterns were found among the Page serovar C isolates, i.e. isolates which reacted with a 1 in 100 dilution of the Page serovar C antiserum, and another larger group which did not react with this dilution of the serum, but did react with rabbit raised antiserum prepared against other Kume serogroup C isolates. This is the first recorded isolation of Page serovar C NAD-independent H. paragallinarum.