ARDRA studies of the ribosomal RNA operon within the Desulfovibrio desulfuricans strains

• Authors: Joanna Szczerba , Ilona Bednarek , Zofia Dzierżewicz
• Languages of publication: EN

Abstracts

EN

BACKGROUND Desulfovibrio desulfuricans belong to the heterogeneous group of anaerobic, sulphate-reducing bacteria (SRB), widely distributed in various environments. As a result of dissimilatory sulphate reduction, they release hydrogen sulphide (H2S), which has a cytotoxic effect in human and animal organisms. It has been shown by many authors, that Desulfovibrio was the genus predominating in patients with ulcerative colitis. Some of these bacteria can act as opportunistic pathogens associated with primary bacteremia and abdominal infections such as abscesses. MATERIAL AND METHODS Fifteen (soil and intestinal) strains of Desulfovibrio desulfuricans species were cultured in modified sulphate-free Postgate’s liquid medium with pyruvate for 10 days. Bacterial DNA was extracted by using a commercially available kit and DNA was used as a template for amplification of the full-length 16S, 23S rDNA and the intergenic spacer region. Digested with restriction enzymes (AluI, EcoRI, HaeIII, HindIII, HinfI, MboI and PstI) PCR amplicons were resolved by electrophoresis on 2% agarose gels. RESULTS Digestion of rrn operon of Desulfovibrio desulfuricans by seven restriction enzymes allowed to obtain the characteristic restriction profiles for all 15 investigated strains. The results allow us to suggest three of used enzymes: HinfI, AluI and HaeIII as a useful for confirmation of the similarity within of rrn operon of isolates belonging to this species. Considering the restriction profiles received with HindIII, and EcoR1 enzymes it seems that their application is insufficient, but PstI enzyme is not acceptable for the analysis of rrn operon of these bacteria. CONCLUSIONS The obtained data have shown that ARDRA can be used for establishment of phylogenetic relations among isolates of Desulfovibrio desulfuricans species, providing the appropriate restriction enzyme is used.

PL

WSTĘP: Bakterie Desulfovibrio desulfuricans należą do szerokiej grupy beztlenowych bakterii redukujących siarczany (BRS), rozpowszechnionej w różnych środowiskach. Jako rezultat dysymilacyjnej redukcji siarczanów uwalniają do środowiska siarkowodór (H2S), który wpływa cytotoksycznie na organizm ludzi i zwierząt. Wielu autorów wykazało, iż rodzaj Desulfovibrio jest dominujący u osób cierpiących z powodu wrzodziejącego zapalenia okrężnicy. Ponadto niektóre gatunki tego rodzaju odgrywają role oportunistycznych patogenów wywołując bakteriemię, infekcje w obrębie jamy brzusznej oraz wrzody. MATERIAŁY I METODY: Piętnaście szczepów Desulfovibrio desulfuricans (glebowych i jelitowych) hodowanych było na modyfikowanym podłożu Postgata z dodatkiem pirogronianu przez 10 dni. Po wyizolowaniu genomowego DNA tych bakterii przeprowadzono reakcje PCR w celu namnożenia fragmentu operonu rrn obejmującego geny 16S, 23S oraz odcinek zmienny pomiędzy nimi. Otrzymane amplikony poddane zostały trawieniu enzymami restrykcyjnymi (AluI, EcoRI, HaeIII, HindIII, HinfI, MboI and PstI), a otrzymane fragmenty rozdzielono w 2% żelu agarozowym. REZULTATY: Przeprowadzona analiza restrykcyjna operonu rrn bakterii Desulfovibrio desulfuricans pozwoliła na otrzymanie charakterystycznych profili restrykcyjnych dla wszystkich piętnastu badanych szczepów. Uzyskane rezultaty pozwoliły uznać trzy z zastosowanych enzymów (HinfI, AluI oraz HaeIII) za odpowiednie do potwierdzenia podobieństw w obrębie operonu rrn bakterii tego gatunku. Biorąc pod uwagę profile otrzymane po trawieniu enzymami HindIII, EcoR1, możemy stwierdzić ich małą użyteczność w analizach operonu rrn gatunku Desulfovibrio desulfuricans, zaś enzym PstI w ogóle nie nadaje się do tego celu.

Keywords

EN

Desulfovibrio desulfuricans; restriction enzymes; ribotyping; sulphate-reducingbacteria

Discipline

• MEDICINE: MEDICINE

• Publisher: Śląski Uniwersytet Medyczny w Katowicach
• Journal: Annales Academiae Medicae Silesiensis
• Year: 2009
• Volume: 63
• Issue: 1
• Pages: 7-13

Contributors

author

Joanna Szczerba

• Department of Biophaemacy Medical University of Silesia, 41-200 Sosnowiec, Poland, Narcyzów 1 St., tel. [phone]

author

Ilona Bednarek

• Department of Biotechnology and Genetic Engineering, Medical University of Silesia, Poland

author

Zofia Dzierżewicz

• Departmant of Biopharmacy, Medical University of Silesia, Poland

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• Document Type: article