ABSTRACT
Biofilm-forming bacteria are highly resistant to antibiotics, host immune defenses, and other external conditions. The formation of biofilms plays a key role in colonization and infection. To explore the mechanism of biofilm formation, mutant strains of Proteus vulgaris XC 2 were generated by Tn5 random transposon insertion. Only one biofilm defective bacterial species was identified from among 500 mutants. Inactivation of the glpC gene coding an anaerobic glycerol-3-phosphate dehydrogenase subunit C was identified by sequence analysis of the biofilm defective strain. Differences were detected in the growth phenotypes of the wild-type and mutant strains under pH, antibiotic, and organic solvent stress conditions. Furthermore, we observed an increase in the phagocytosis of the biofilm defective strain by the mouse macrophage RAW264.7 cell line compared to the wild-type strain. This study shows that the glpC gene plays an important role in biofilm formation, in addition to imparting pH, organic solvent, and antibiotic tolerance, and defense against phagocytosis to Proteus sp. The results further clarified the mechanism of biofilm formation at the genomic level, and indicated the importance of the glpC gene in this process. This data may provide innovative therapeutic measures against P. vulgaris infections; furthermore, as an important crocodile pathogen, this study also has important significance in the protection of Chinese alligators.
Subject(s)
Bacterial Proteins/genetics , Biofilms/growth & development , Glycerolphosphate Dehydrogenase/genetics , Proteus Infections/veterinary , Proteus vulgaris/genetics , Proteus vulgaris/immunology , Adaptation, Physiological/immunology , Alligators and Crocodiles/microbiology , Animals , Bacterial Proteins/immunology , Biofilms/drug effects , Cell Line , Cyclohexanes/pharmacology , DNA Transposable Elements , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Expression , Glycerolphosphate Dehydrogenase/immunology , Hexanes/pharmacology , Hydrogen-Ion Concentration , Immune Evasion , Macrophages/microbiology , Mice , Mutation , Proteus Infections/microbiology , Proteus Infections/pathology , Proteus vulgaris/drug effects , Proteus vulgaris/isolation & purification , Recombinant Proteins/genetics , Recombinant Proteins/immunologyABSTRACT
A clinical isolate of Proteus mirabilis (Pr 990) and an isogenic non-flagellate allelic replacement mutant (Pr M9) were tested for their ability to cause infection in the ascending mouse model of urinary tract infection. Wild-type Pr 990 differentiates into swarmer cells in brain-heart infusion broth. Pr M9 neither has flagella nor does it apparently differentiate into swarmer cells after subculturing. The infectivity of both strains from an initial culture and the sixth subculture was assessed in the ascending urinary tract infection mouse model. Infection was ascertained by determining colony forming units from kidney and bladder homogenates from individual mice 7 days after inoculation. In all cases the nonflagellate mutant Pr M9 was at least as infective as Pr 990. Using bacteria from the first culture, Pr M9 infected 61.5% and Pr 990 infected 45.5% of mice tested. The levels of viable counts were similar between the Pr M9 and the Pr 990 infections. Using bacteria from the sixth subculture, Pr M9 infected 75% and Pr 990 infected 76.5% of mice tested. Again viable counts were similar. Pr 990 increased in infectivity from the first to the sixth subculture, whereas Pr M9 did not, but this may be a reflection of the high initial rate of infectivity with first culture Pr M9. These results suggest that neither flagella nor swarmer cells are required for P. mirabilis infectivity in ascending urinary tract infections in mice.
Subject(s)
Flagellin/metabolism , Proteus Infections/microbiology , Proteus mirabilis/physiology , Urinary Tract Infections/microbiology , Alleles , Animals , Cattle , Chickens , Cloning, Molecular , Disease Models, Animal , Female , Flagella/genetics , Flagellin/genetics , Guinea Pigs , Horses , Humans , Mice , Mutagenesis , Phenotype , Proteus Infections/pathology , Proteus mirabilis/genetics , Sheep , Urinary Tract Infections/pathologyABSTRACT
Dermatological lesions of a patient are studied and investigated. It is suggested that they belong to a syndrome rather than to an entity. The following etiological classification of the mossy syndrome is proposed: due to bacteria, fungi and viruses, and secondary to other dermatopathies. Bibliography is reviewed.