title: Actinobacillus pleuropneumoniae serotype 1 carrying the defined aroA mutation is fully avirulent in the pig.
abstract: The aroA gene from Actinobacillus pleuropneumoniae serotype 1 reference strain 4074 was isolated and sequenced. The gene complemented the aroA mutation in Escherichia coli AB2829. A kanamycin resistance cassette was inserted into the aroA gene and the mutant gene was reintroduced into A. pleuropneumoniae by allelic replacement. Intratracheal infection of susceptible pigs with A. pleuropneumoniae aroA caused no signs of respiratory disease or lung lesions in any of the animals at a dose 10(4) times the dose reliably known to induce acute pleuropneumonia; all animals infected with the unaltered control strain developed acute disease. The aroA mutant was rapidly eliminated from the lungs and tonsil of infected animals. The mutant may represent a safely attenuated strain for use in live bacterial vaccination or the delivery of antigen by the intranasal route. However, the residence time of the mutant in the respiratory tract of the pig may be too short for it to be useful in generating a protective immune response.
authors.affiliation: Veterinary Bacteriology Group, Department of Pathology and Infectious Disease, Royal Veterinary College, Hawkshead Lane, North Mymms, Herts, AL9 7TA, UK.
authors.forename: L H
authors.initials: LH
authors.lastname: Garside
authors.forename: M
authors.initials: M
authors.lastname: Collins
authors.forename: P R
authors.initials: PR
authors.lastname: Langford
authors.forename: A N
authors.initials: AN
authors.lastname: Rycroft
mesh_terms: D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D000189:Actinobacillus Infections / Q000382:microbiology / Q000662:veterinary; D016977:Actinobacillus pleuropneumoniae / Q000145:classification / Q000201:enzymology / Q000235:genetics* / Q000472:pathogenicity*; D019883:Alkyl and Aryl Transferases / Q000235:genetics / Q000378:metabolism*; D000483:Alleles; D000818:Animals; D015870:Gene Expression; D018014:Gene Transfer Techniques; D007612:Kanamycin; D007613:Kanamycin Resistance / Q000235:genetics; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D013552:Swine / Q000382:microbiology*; D014774:Virulence / Q000235:genetics
chemical_list: D007612:Kanamycin; D019883:Alkyl and Aryl Transferases; D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase