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Journal of Endotoxin Research
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Identification of genes involved in biosynthesis of Actinobacillus pleuropneumoniae serotype 1 O-antigen and biological properties of rough mutants

Josée Labrie

Groupe de recherche sur les maladies infectieuses du porc, Département de pathologie et microbiologie, Faculté de médecine vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada

Stéphane Rioux

Groupe de recherche sur les maladies infectieuses du porc, Département de pathologie et microbiologie, Faculté de médecine vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada, Unité de recherche en vaccinologie, Centre Hospitalier Universitaire de Québec, Pavillon CHUL, Sainte-Foy, Québec, Canada, G1V 4G2

Mary Margaret Wade

Department of Biological Sciences, Mississippi State University, Mississippi State, Mississippi, USA

Franklin R. Champlin

Department of Biological Sciences, Mississippi State University, Mississippi State, Mississippi, USA

Steven C. Holman

Department of Chemistry, Mississippi State University, Mississippi State, Mississippi, USA

W. William Wilson

Department of Chemistry, Mississippi State University, Mississippi State, Mississippi, USA

Chantal Savoye

Agence Française de Sécurité Sanitaire des Aliments, Laboratoire d'études et de recherches avicoles et porcines, Unité de recherche Mycoplasmologie Bactériologie, Zoopôle, Ploufragan, France

Marylène Kobisch

Agence Française de Sécurité Sanitaire des Aliments, Laboratoire d'études et de recherches avicoles et porcines, Unité de recherche Mycoplasmologie Bactériologie, Zoopôle, Ploufragan, France

Marc Sirois

Département de chimie-biologie, Université du Québec à Trois-Rivières, Trois-Rivières, Québec, Canada

Catherine Galarneau

Groupe de recherche sur les maladies infectieuses du porc, Département de pathologie et microbiologie, Faculté de médecine vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada

Mario Jacques

Groupe de recherche sur les maladies infectieuses du porc, Département de pathologie et microbiologie, Faculté de médecine vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada, mario.jacques{at}umontreal.ca

Actinobacillus pleuropneumoniae is an important pathogen of swine. Lipopolysaccharide (LPS) has been identified as the major adhesin of A. pleuropneumoniae and it is involved in adherence to porcine respiratory tract cells. We previously generated seven rough LPS mutants of A. pleuropneumoniae serotype 1 by using a mini-Tn10 transposon mutagenesis system [Rioux S, Galarneau C, Harel J et al. Isolation and characterization of mini-Tn10 lipopolysaccharide mutants of Actinobacillus pleuropneumoniae serotype 1. Can J Microbiol 1999; 45: 1017—1026]. The purpose of the present study was to characterize these mutants in order to learn more about LPS O-antigen biosynthesis genes and their organization in A. pleuropneumoniae, and to determine the surface properties and virulence in pigs of these isogenic mutants. By mini-Tn10 insertions in rough mutants, four putative genes (ORF12, ORF16, ORF17, and ORF18) involved in O-antigen biosynthesis in A. pleuropneumoniae serotype 1 were found within a region of 18 ORFs. This region is homologous to the gene cluster of serotype-specific O-polysaccharide biosynthesis from A. actinomycetemcomitans strain Y4 (serotype b). Two mutants showed homology to a protein with identity to glycosyltransferases (ORF12); two others had the mini-Tn10 insertion localized in genes encoding for two distinct proteins with identity to rhamnosyltransferases (ORF16 and ORF17) and three showed homology to a protein which is known to initiate polysaccharide synthesis (ORF18). These four ORFs were also present in A. pleuropneumoniae serotypes 9 and 11 that express an O-antigen that serologically cross-reacts with serotype 1. Evaluation of some biological properties of rough mutants seems to indicate that the absence of O-chains does not appear to have an influence on the virulence of the bacteria in pigs and on the overall surface hydrophobicity, charge and hemoglobin-binding activity, or on LAL activation. An acapsular mutant was included in the present study in order to compare the influence of O-chains and capsule polysaccharides on different cell surface properties. Our data suggest that capsular polysaccharides and not O-chains polysaccharides have a major influence on surface properties of A. pleuropneumoniae serotype 1 and its virulence in pigs.

Journal of Endotoxin Research, Vol. 8, No. 1, 27-38 (2002)
DOI: 10.1177/09680519020080010301
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