Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
JCR Impact Factor (IF) – 2.1
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ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
Periodicity – monthly

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Advances in Clinical and Experimental Medicine

2010, vol. 19, nr 6, November-December, p. 669–677

Publication type: original article

Language: English

The Sensitivity Level of Salmonella Enterica ESBL+ Transconjugants to Normal Human Serum Correlated with OMP Band Patterns Obtained by SDS-PAGE

Poziom wrażliwości transkoniugantów Salmonella enterica ESBL+ na działanie normalnej surowicy ludzkiej w korelacji z profilami OMP uzyskanymi metodą SDS-PAGE

Jolanta Sarowska1,, Gabriela Bugla-Płoskońska2,, Bożena Futoma-Kołoch2,, Zuzanna Drulis-Kawa3,

1 Department of Biology and Medical Parasitology, Wroclaw Medical University, Wrocław, Poland

2 Institute of Genetics and Microbiology, University of Wroclaw, Wrocław, Poland

3 Department of Pathogen Biology and immunology, University of Wroclaw, Wrocław, Poland

Abstract

Objectives. The aim of the study was to assess the susceptibility of Salmonella strains to the bactericidal activity of normal human serum (NHS) and to analyze the outer membrane protein (OMP) profiles.
Material and Methods. The clinical strain Klebsiella pneumoniae 20 and six Salmonella enterica strains – S. Enteritidis 518, S. Typhimurium 244 and S. Hadar 55 and their ESBL+ transconjugants S. Enteritidis 518/20, S. Typhimurium 244/20 and S. Hadar 55/20, obtained with K. pneumoniae 20 as the donor of ESBL plasmid were used. The bactericidal activity of NHS was determined as described by Jankowski et al. Isolation of the OMPs was performed according to the procedure described by Murphy and Bartos. Discontinuous sodium dodecyl sulphate (SDS) gel electrophoresis was carried out as described by Laemmli.
Results. This study examined the susceptibility of Salmonella recipient strains and their ESBL+ transconjugants belonging to three serovars – Enteritidis, Typhimurium and Hadar – to NHS. It was shown that the recipient strains (S. Enteritidis 518, S. Typhimurium 244, and S. Hadar 55) and the ESBL conjugative plasmid donor strain (K. pneumoniae 20) were resistant to the bactericidal action of NHS, whereas three Salmonella transconjugants identified as ESBL producers (S. Enteritidis 518/20, S. Typhimurium 244/20 and S. Hadar 55/20) demonstrated high sensitivity to serum. SDS-PAGE gel analysis showed that the parental strains and transconjugants exhibited different outer membrane protein patterns. The observed changes included the presence or absence of particular proteins.
Conclusion. It seems that the acquisition of new plasmids may have unfavorable consequences for pathogenic bacteria and increase their susceptibility to serum activity.

Streszczenie

Cel pracy. Określenie podatności szczepów Salmonella na bakteriobójcze działanie normalnej surowicy ludzkiej (NHS) i analiza profili białek błony zewnętrznej (OMP) badanych szczepów.
Materiał i metody. W badaniach wykorzystano kliniczny szczep Klebsiella pneumoniae 20 oraz sześć izolatów Salmonella enterica: S. Enteritidis 518, S. Typhimurium 244, S. Hadar 55, a także ich ESBL+ transkoniuganty: S. Enteritidis 518/20, S. Typhimurium 244/20 i S. Hadar 55/20, otrzymane z użyciemu szczepu K. pneumoniae 20, jako dawcy plazmidu typu ESBL. Bakteriobójczą aktywność NHS oceniano metodą opisaną przez Jankowskiego et al. Izolację białek OMP przeprowadzono zgodnie z procedurą opisaną przez Murphy’ego i Bartosa. Elektroforezę białek na żelu poliakrylamidowym w obecności SDS wykonano w układzie Laemmli’ego.
Wyniki. W badaniach określono wrażliwość na działanie NHS wyjściowych szczepów biorców Salmonella oraz ich ESBL+ transkoniugantów należących do trzech serowarów: Enteritidis, Typhimurium i Hadar. Wykazano, że zarówno szczepy biorców (S. Enteritidis 518, S. Typhimurium 244, S. Hadar 55), jak i szczep dawcy plazmidu typu ESBL (K. pneumoniae 20) były oporne na bakteriobójcze działanie NHS, a trzy uzyskane transkoniuganty ESBL+ Salmonella (S. Enteritidis 518/20, S. Typhimurium 244/20, S. Hadar 55/20) okazały się bardzo wrażliwe na działanie surowicy. Rozdział SDS-PAGE wykazał różnice na poziomie OMP między szczepami wyjściowymi Salmonella, a ich ESBL+ transkoniugantami. Obserwowane zmiany polegały na obecności lub braku poszczególnych białek. Wniosek. Prawdopodobnie pozyskanie nowego plazmidu przez komórki bakteryjne wpłynęło na niekorzystny dla tych patogenów wzrost podatności na aktywność surowicy.

Key words

Salmonella, NHS, OMP

Słowa kluczowe

Salmonella, NHS, OMP

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