Isolation and characterization of a capsule-deficient mutant of Actinobacillus pleuropneumoniae serotype 1.
The capsular polysaccharides (CPS) play a major role in pathogenicity of Actinobacillus pleuroIpneumoniae, the causative agent of porcine pleuropneumonia. The purpose of the present study was to isolate a mutant in CPS biosynthesis by using a mini-Tn 10 transposon mutagenesis system and evaluate its adherence to host cells. One mutant apparently did not possess CPS as it did not react with a monoclonal antibody against A. pleuropneumoniae serotype 1 capsular antigen. Absence of capsule was confirmed by flow cytometry and also by transmission electron microscopy after polycationic ferritin labelling. The site of insertion of the mini-Tn 10 was determined and found to be in the cpxC gene. Its gene product, CpxC, is a protein involved in polysaccharide transport across the cytoplasmic membrane during CPS biosynthesis. Use of piglet tracheal frozen sections indicated that the CPS mutant adhered significantly (P=0.0001) more than the parent strain. The non-capsular mutant was less virulent in pigs compared to the parent strain and showed no mortality in experimentally infected pigs. The CPS mutant was however resistant to pig serum. This CPS mutant is the first A. pleuropneumoniae mutant in a CPS transport gene. It is also the first time that adherence of a CPS mutant of A. pleuropneumoniae is evaluated. Our observations indicate that capsular polysaccharides of A. pleuropneumoniae serotype 1 are not involved in adherence to piglet tracheal frozen sections but rather mask, at least in part, the adhesive functions.
Rioux
S
S
Groupe de Recherche sur les Maladies Infectieuses du Porc, and Département de Pathologie et Microbiologie, Université de Montréal, St-Hyacinthe, Québec, J2S 7C6, Canada.
Galarneau
C
C
Harel
J
J
Kobisch
M
M
Frey
J
J
Gottschalk
M
M
Jacques
M
M
D000189:Actinobacillus Infections / Q000382:microbiology / Q000401:mortality / Q000473:pathology / Q000662:veterinary*; D016977:Actinobacillus pleuropneumoniae / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D000818:Animals; D000911:Antibodies, Monoclonal; D001422:Bacterial Adhesion; D016667:Bacterial Capsules / Q000235:genetics*; D004251:DNA Transposable Elements; D005434:Flow Cytometry; D015151:Immunoblotting; D008070:Lipopolysaccharides / Q000032:analysis; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D012703:Serotyping; D013552:Swine; D013553:Swine Diseases / Q000382:microbiology* / Q000401:mortality / Q000473:pathology; D014132:Trachea / Q000382:microbiology / Q000473:pathology; D014774:Virulence
D000911:Antibodies, Monoclonal; D004251:DNA Transposable Elements; D008070:Lipopolysaccharides