Molecular analysis of clinical isolates of Providencia alcalifaciens

Sobreira, Marise; Leal, Nilma Cintra; Magalhães, Maria João; Guth, Beatriz E. C.; Almeida, Alzira Maria Paiva de

doi:10.1099/0022-1317-50-1-29

Cited by 18 publications

(10 citation statements)

References 11 publications

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“…Among these, P. alcalifaciens has been described as a causative agent of diarrhea because a number of P. alcalifaciens strains were isolated from patients with diarrhea in developing countries (21,22,47,48). Indeed, a case control study conducted by Albert et al (3) demonstrated that P. alcalifaciens was associated with diarrhea in children in Bangladesh.…”

Section: Discussionmentioning

confidence: 99%

“…Invasion was also observed in intestinal tissues by using a removable intestinal tie adult rabbit diarrhea (RITARD) model and an adult rabbit ileal loop model (1,35). Although invasion was considered as one of the virulence mechanisms to cause diarrhea by P. alcalifaciens strains, noninvasive P. alcalifaciens were also isolated from patients with diarrhea (21,48). In our study, one strain, P. alcalifaciens AH-31, showed invasiveness to HeLa cells; however, another strain, P. alcalifaciens strain AS-1, did not show any invasiveness (data not shown).…”

Section: Discussionmentioning

confidence: 99%

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Molecular Characterizations of Cytolethal Distending Toxin Produced by Providencia alcalifaciens Strains Isolated from Patients with Diarrhea

et al. 2012

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Cytolethal distending toxins (CDTs), which block eukaryotic cell proliferation by acting as inhibitory cyclomodulins, are produced by diverse groups of Gram-negative bacteria. Active CDT is composed of three polypeptides-CdtA, CdtB, and CdtC-encoded by the genes cdtA, cdtB, and cdtC, respectively. We developed a PCR-restriction fragment length polymorphism assay for the detection and differentiation of five alleles of cdtB (Cdt-I through Cdt-V) in Escherichia coli and used the assay to investigate the prevalence and characteristic of CDT-producing E. coli in children with diarrhea (A. Hinenoya et al., Microbiol. Immunol. 53:206 -215, 2009). In these assays, two untypable cdtB genes were detected and the organisms harboring the cdtB gene were identified as Providencia alcalifaciens (strains AH-31 and AS-1). Nucleotide sequence analysis of the cdt gene cluster revealed that the cdtA, cdtB, and cdtC genes of P. alcalifaciens are of 750, 810, and 549 bp, respectively. To understand the possible horizontal transfer of the cdt genes among closely related species, the presence of cdt genes was screened in various Providencia spp. by colony hybridization assay, and the cdt gene cluster was found in only limited strains of P. alcalifaciens. Genome walking revealed that the cdt gene cluster of P. alcalifaciens is located adjacent to a putative transposase gene, suggesting the locus might be horizontally transferable. Interestingly, the CDT of P. alcalifaciens (PaCDT) showed some homology with the CDT of Shigella boydii. Whereas filter-sterilized lysates of strains AH-31 and AS-1 showed distention of CHO but not of HeLa cells, E. coli CDT-I exhibited distention of both cells. This activity of PaCDT was confirmed by generating recombinant PaCDT protein, which could also be neutralized by rabbit anti-PaCdtB antibody. Furthermore, recombinant PaCDT was found to induce G 2 /M cell cycle arrest and phosphorylation of host histone H2AX, a sensitive marker of DNA double-strand breaks. To our knowledge, this is the first report showing that certain clinical P. alcalifaciens strains could produce variants of the CDTs compared.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Molecular Characterizations of Cytolethal Distending Toxin Produced by Providencia alcalifaciens Strains Isolated from Patients with Diarrhea

et al. 2012

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“…Providencia rettgeri, Providencia alcalifaciens, and Providencia stuartii have all been isolated from human stool samples both as part of the natural human gut flora and as the cause of gastric upset such as traveler’s diarrhea [16–18]. Some strains of P. alcalifaciens , but no strains of P. rettgeri or P. stuartii , have been found to be intracellularly invasive in human cell lines [16, 17, 19, 20]. Providencia also cause urinary tract and other nosocomial infections in humans [12, 13].…”

Section: Introductionmentioning

confidence: 99%

Comparative pathology of bacteria in the genus Providencia to a natural host, Drosophila melanogaster

Galac

Lazzaro

2011

Microbes and Infection

120

137

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Bacteria in the genus Providencia are pathogens of many organisms, including humans and insects. We and colleagues have isolated five different strains belonging to four distinct Providencia species as natural infections of Drosophila melanogaster captured in the wild. We found that these isolates vary considerably in pathology to infected D. melanogaster, differing in the level of mortality they cause, their ability to replicate within the host and the level that the fly’s immune response is elicited. One interesting bacterium was Providencia sneebia, which causes nearly complete mortality and reaches large numbers in the fly but does not elicit a comparably strong immune response. Through coinfection experiments, we determined that P. sneebia avoids recognition by the immune system. We tested for biofilm formation and replication within D. melanogaster cells as possible mechanisms for P. sneebia escape from host immunity, but did not find evidence for either. D. melanogaster and Providencia provide a powerful system for studying general host-pathogen interactions, and for understanding how the well-studied immune model host D. melanogaster interacts with its natural bacterial pathogens.

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“…Invasion of epithelial cells in vitro is associated with condensation of actin filaments, and both invasion and actin condensation can be inhibited by cytochalasin D, an inhibitor of microfilament formation [6]. Even though some isolates of P. alcalifaciens carry plasmids, isolates with and without plasmids were found to be equally invasive, indicating that chromosomal genes, not plasmid genes, are involved in invasion [8]. Also, no homologous sequence corresponding to virulence genes of invasive bacteria such as Shigella spp., Yersinia enterocolitica and Y. pseudotuberculosis could be found [8].…”

Section: Introductionmentioning

confidence: 99%

“…Even though some isolates of P. alcalifaciens carry plasmids, isolates with and without plasmids were found to be equally invasive, indicating that chromosomal genes, not plasmid genes, are involved in invasion [8]. Also, no homologous sequence corresponding to virulence genes of invasive bacteria such as Shigella spp., Yersinia enterocolitica and Y. pseudotuberculosis could be found [8]. In many bacteria, genes encoding surface-exposed or secreted products can be identified by TnphoA mutagenesis.…”

Section: Introductionmentioning

confidence: 99%

TnphoA mutants of Providencia alcalifaciens with altered invasiveness of HEp-2 cells

Rahman¹,

Monira

Nahar³

et al. 2002

Journal of Medical Microbiology

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Molecular analysis of clinical isolates of Providencia alcalifaciens

Cited by 18 publications

References 11 publications

Molecular Characterizations of Cytolethal Distending Toxin Produced by Providencia alcalifaciens Strains Isolated from Patients with Diarrhea

Molecular Characterizations of Cytolethal Distending Toxin Produced by Providencia alcalifaciens Strains Isolated from Patients with Diarrhea

Comparative pathology of bacteria in the genus Providencia to a natural host, Drosophila melanogaster

TnphoA mutants of Providencia alcalifaciens with altered invasiveness of HEp-2 cells

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