RESUMO
The occurrence of bacteria belonging to the family Pasteurellaceae in the oral cavity of captive Tasmanian devils (Sarcophilus harrisii) was investigated using phenotypic and subsequent genotypic characterization and phylogenetic analyses. A total of 62 bacterial isolates obtained from Tasmanian devils, tentatively classified with the family Pasteurellaceae, were further characterized by phylogenetic analysis of rpoB gene sequence similarity, which showed that the isolates investigated formed five distinct groups. A total of 15 strains formed a novel genus-like group within Pasteurellaceae. Thirty-six strains grouped with the type strain of Frederiksenia canicola. Five strains clustered with the type strain of Pasteurella multocida . Interestingly, four of the P. multocida-like strains were ß-hemolytic when incubated on blood agar, which is atypical for this genus. Five strains grouped with a 100% rpoB similarity with Pasteurella dagmatis. Finally, a single strain showed 97.1% resemblance to Haemophilus haemoglobinophilus. The results demonstrate that Tasmanian devils are hosting a variety of bacterial taxa affiliated with the family of Pasteurellaceae as part of their oral microflora.
Assuntos
Marsupiais/microbiologia , Boca/microbiologia , Pasteurellaceae/isolamento & purificação , Animais , Animais de Zoológico , Pasteurellaceae/genética , FilogeniaRESUMO
The phage-shock protein (Psp) system is believed to manage membrane stress in all Enterobacteriaceae and has recently emerged as being important for virulence in several pathogenic species of this phylum. The core of the Psp system consists of the pspA-D operon and the distantly located pspG gene. In Salmonella enterica serovar Typhimurium (S. Typhimurium), it has recently been reported that PspA is essential for systemic infection of mice, but only in NRAMP1(+) mice, signifying that attenuation is related to coping with divalent cation starvation in the intracellular environment. In the present study, we investigated the contribution of individual psp genes to virulence of S. Typhimurium. Interestingly, deletion of the whole pspA-D set of genes caused attenuation in both NRAMP1(+) and NRAMP1(-) mice, indicating that one or more of the psp genes contribute to virulence independently of NRAMP1 expression in the host. Investigations of single gene mutants showed that knock out of pspB reduced virulence in both types of mice, while deletion of pspA only caused attenuation in NRAMP1(+) mice, and deletion of pspD had a minor effect in NRAMP1(-) mice, while deletions of either pspC or pspG did not affect virulence. Experiments addressed at elucidating the role of PspB in virulence revealed that PspB is dispensable for uptake to and intracellular replication in cultured macrophages and resistance to complement-induced killing. Furthermore, the Psp system of S. Typhimurium was dispensable during pIV-induced secretin stress. In conclusion, our results demonstrate that removal of PspB reduces virulence in S. Typhimurium independently of host NRAMP1 expression, demonstrating that PspB has roles in intra-host survival distinct from the reported contributions of PspA.