Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
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Advances in Clinical and Experimental Medicine

2009, vol. 18, nr 1, January-February, p. 63–70

Publication type: original article

Language: English

Conjugative Transfer of Plasmid Encoding Extended−Spectrum Beta−Lactamase to Recipient Salmonella Strains

Koniugacyjny transfer plazmidu kodującego geny odpowiedzialne za syntezę β−laktamaz o rozszerzonym spektrum substratowym do komórek biorców Salmonella sp.

Jolanta Sarowska1,, Zuzanna Drulis−Kawa2,, Katarzyna Guz2,, Stanislaw Jankowski1,, Dorota Wojnicz1,

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

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

Abstract

Background. Salmonella enterica subsp. enterica is one of the most frequently occurring etiological factors of severe gastroenteritis in children. Young children are also commonly colonized by bacterial strains producing ESBL enzymes. These enzymes are encoded on plasmids and can be easy transferred by conjugation to other Gram−negative strains, especially from fecal flora.
Objectives. This study examined the transferability of ESBL plasmid from Klebsiella pneumoniae strains to clinical Salmonella isolates and then between clinical Salmonella strains, which leads to Salmonella resistance to betalactam antibiotics, the most often used in treatment.
Material and Methods. The transferability of β−lactam resistance determinants was studied by the bacterial conjugation method. K. pneumoniae ATCC 700603 carrying SHV−18 ESBL genes and a clinical isolate of K. pneumoniae 20 (ESBL+) were used as the donors of ESBL plasmid. S. enterica serovar Enteritidis (n = 10), S. enterica serovar Typhimurium (n = 10), and S. enterica serovar Hadar (n = 10) isolates were used as the recipient strains. Before and after conjugation the MICs of amikacin (AK), ampicillin (AM), ceftriaxone (TX), ceftazidime (TZ), trimethoprim−sulfametoksazol (TS), and ciprofloxacin (CI) were measured by the E−test. All isolates were tested for ESBL activity by the disc diffusion test (the Jarlier Double−Disc test) and the E−test. The plasmid DNA patterns were analyzed by the alkaline method.
Results. All S. Enteritidis (10/10), several S. Typhimurium (4/10), and one S. Hadar (1/10) isolate became transconjugants. All Salmonella transconjugants were identified as ESBL producers. After conjugation, increases in the MICs of AM, TX, and TZ were observed. Conclusion. The efficiency of ESBL plasmid transferability depended on the Salmonella serovar. The S. enterica serovar Enteritidis strains were the best ESBL plasmid recipients among the clinical Salmonella serovars.

Streszczenie

Wprowadzenie. Pałeczki Salmonella enterica subsp. enterica są jednym z najczęstszych czynników etiologicznych gastroenteritis u dzieci. Przewód pokarmowy dzieci jest często kolonizowany przez szczepy bakteryjne wytwarzające enzymy typu ESBL, których genetyczne determinanty zlokalizowane w plazmidach mogą być łatwo przekazywane na drodze koniugacji, szczególnie w obrębie bakterii Gram−ujemnych.
Cel pracy. Określenie częstości koniugacyjnego transferu plazmidów typu ESBL ze szczepów Klebsiella pneumoniae do komórek szczepów należących do przedstawicieli trzech serowarów Salmonella: Enteritidis, Typhimurium i Hadar oraz w obrębie pałeczek Salmonella.
Materiał i metody. Częstość przekazywania plazmidów typu ESBL określono metodą koniugacji w podłożu bulionowym. Jako dawców plazmidu typu ESBL zastosowano szczepy K. pneumoniae ATCC 700603 (SHV−18) i K. pneumoniae 20 (ESBL+). Szczepy S. enterica serowar Enteritidis (n = 10), S. enterica serowar Typhimurium (n = 10) i S. enterica serowar Hadar (n = 10) wykorzystano jako biorców. Wartości MIC dla amikacyny (AK), ampicyliny (AM), ceftriaksonu (TX), ceftazydymu (TZ), trimetoprimu z sulfametoksazolem (TS) i ciprofloksacyny (CI) określono metodą E−testów przed i po koniugacji. Zdolność do syntezy enzymów ESBL badano metodą dwóch krążków (DDST) wg Jarliera oraz metodą E−testów. Analizę profili plazmidowego DNA przeprowadzono metodą lizy alkalicznej.
Wyniki. Wprzypadku wszystkich badanych szczepów S. Enteritidis (10/10), czterech S. Typhimurium (4/10) i jednego izolatu S. Hadar (1/10) uzyskano transkoniuganty, które wykazywały produkcję beta−laktamaz o rozszerzonym spektrum substratowym (ESBL). Zaobserwowano wzrost wartości MIC dla AM, TX i TZ wobec uzyskanych transkoniugantów. Wniosek. Wyniki badań wskazują, że skuteczność i częstość koniugacyjnego transferu plazmidu typu ESBL była różna dla poszczególnych serowarów Salmonella. Szczepy S. Enteritidis charakteryzowały się największą zdolnością do uzyskiwania plazmidu ESBL.

Key words

Salmonella, horizontal genes transfer, ESBL

Słowa kluczowe

Salmonella, horyzontalny transfer genów, ESBL

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