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Rupnik, M.

Publications and source records attributed to Rupnik, M..

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Lytic Bacteroides uniformis bacteriophages exhibiting host tropism congruent with diversity generating retroelement

Intestinal phages are abundant and important component of gut microbiota, but our knowledge remains limited to only a few isolated and characterized representatives targeting numerically dominant gut bacteria. Here we describe isolation of human intestinal phages infecting Bacteroides uniformis. Bacteroides is one of the most common bacterial groups in the global human gut microbiota, however, to date not many Bacteroides specific phages are known. Phages isolated in this study belong to a novel viral genus, Bacuni, within Siphoviridae family and represent the first lytic phages, genomes of which encode diversity generating retroelements (DGR). This region is assumed to promote phage adaptation to the rapidly changing environmental conditions and to broaden its host range. Three isolated phages showed 99,83% genome identity but infected distinct B. uniformis strains. The tropism of Bacuni phages appeared to be dependent on the interplay of DGR mediated sequence variations of phage fimbrial tip proteins and mutations in host genes coding for outer-membrane proteins. We found prophages with up to 85% aa similarity to Bacuni phages in the genomes of B. acidifaciens and Prevotella sp.. Despite the abundance of Bacteroides within human microbiome, we found Bacuni phages only in a limited subset of published gut metagenomes. ImportanceThe lack of common marker gene in viruses require a precise characterization of diverse isolated phages to enhance metagenomic analyses and to understand their role in gut microbiota. Here we report the isolation of phages representing a new genus with characteristics so far not known or rarely described in intestinal phages. They are the first lytic phages specific for Bacteroides uniformis, a bacterial representative of the prevalent genus in the gut of humans and animals. Additionally, they are the first lytic phages containing specific regions (diversity generating retroelement) that putatively influence host tropism. The ability to switch constantly the targeted populations of the host species could provide an evolutionary advantage to these bacteriophages and may affect intra species diversity.

microbiology

Comparison between cultivation and sequencing based approaches for microbiota analysis in swabs and biopsies of chronic wounds

Chronic wounds are a prominent health concern affecting 0.2% of individuals in the Western population. Microbial colonization and the consequent infection contribute significantly to the healing process of chronic wounds. We have compared cultivation and 16S amplicon sequencing (16S-AS) for the characterization of bacterial populations in swabs and biopsy tissues obtained from 45 chronic wounds and analysed metadata for wound-specific and clinical-outcome-associated correlations with bacterial community structure. Using cultivation approach, we detected a total of 39 bacterial species, on average 2.89 per sample (SD=1.93). Comparison of cultivation results between swabs and biopsy samples showed no significant advantage of one sampling method over the other. 16S-AS was advantageous in comparison to the cultivation approach in case of highly diverse communities, where we could additionally detect numerous obligate and facultative anaerobic bacteria from genera Anaerococcus, Finegoldia, Porphyromonas, Morganella and Providencia. Based on the community diversity, chronic wound microbiota could be distributed into three groups, however, no correlation between groups and clinical outcome was observed. Clinically estimated presence of biofilm and a larger surface area at the initial visit were most significantly associated with unfavourable clinical outcomes after one-year follow-up visit. Corynebacterium was the single most predictive bacterial genus associated with unfavourable clinical outcomes in our study.

microbiology