Microbial Profile in Bile from Cholecystectomised Patients by Culture and Multiplex PCR

Author:

Farhana Nasreen,Fardows Jannatul,Khan Mohammad Ashraf Uddin,Shamsuzzaman SM

Abstract

AbstractObjectivesBile in the biliary tract is normally sterile but presence of gallstones, ascending infection from duodenum or bacterial translocation from portal vein leads to microfloral colonization. Therefore, this study was aimed to evaluate the microbiological profile of bile for determination of the appropriate antimicrobials in cholecystectomised patients.DesignThis was a descriptive type of cross sectional study from July, 2013 to December, 2014SettingIn a tertiary medical college hospital, Dhaka, BangladeshParticipantsThis study included 246 patients irrespective of age and sex who underwent laparoscopic or open cholecystectomy for various indications in a tertiary hospital, Dhaka, Bangladesh.Primary and secondary outcome measuresSimilar secondary outcome as in protocolMethodsThe intraoperative bile samples were collected and cultured in Blood agar and MacConkey’s agar media aerobically in the laboratory of Microbiology Department, Dhaka Medical College. The isolates were then tested for antibiotic sensitivity pattern. To detect anaerobic bacteria (Bacteroides fragilis, Clostridium perfringens and Fusobacterium nucleatum), multiplex polymerase chain reaction (PCR) was used with specific primers.ResultsOut of 246 bile samples, organisms were identified in 69.51% cases where 48.37% were aerobic bacteria identified by culture and 21.14% were anaerobic bacteria identified by multiplex PCR. Escherichia coli (26.61%) were found predominantly followed by Staphylococcus aureus (19.35%), Citrobacter freundii (14.52%), Clostridium perfringens (13.82%), Pseudomonas spp. (11.29%), Salmonella enterica serovar Typhi 8 (3.45%), and Acinetobacter spp. (3.23%). Gram negative bacteria showed resistance to amoxiclav, ceftriaxone and ceftazidime while meropenem, piperacillin-tazobactam and colistin were found more effective antimicrobials.ConclusionsA great proportion of aerobic and anaerobic bacterial infection associated with gallstones or biliary tract obstruction may warrants serious health risk to cholecystectomised patients in this region. Regular surveillance of bile culture from gallbladder should be done after cholecystectomy.StrengthThe antimicrobial susceptibility pattern of aerobic bacteria was assessed to find out multidrug resistant bacteriaMultiplex PCR was done to detect important biliary anaerobic bacteriaLimitationsIsolation by culture and the antimicrobial susceptibility pattern of anaerobic bacteria was not assessedDue to lack of primers, PCR could not be done for other biliary anaerobic bacteria, virulence genes and drug resistance genesSource(s) of funding for workThis research received no specific grant from any funding agency in the public, commercial or not-for-profit sectors.Competing interestsNoneConflict of interestsNone

Publisher

Cold Spring Harbor Laboratory

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