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Association of diverse bacterial communities in human bile samples with biliary tract disorders: a survey using culture and polymerase chain reaction-denaturing gradient gel electrophoresis methods

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Abstract

Bacterial infection is considered a predisposing factor for disorders of the biliary tract. This study aimed to determine the diversity of bacterial communities in bile samples and their involvement in the occurrence of biliary tract diseases. A total of 102 bile samples were collected during endoscopic retrograde cholangiopancreatography (ERCP). Characterization of bacteria was done using culture and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) methods. Antimicrobial susceptibility of the isolates was determined based on the Clinical and Laboratory Standards Institute (CLSI) guidelines and identity of the nucleotide sequences of differentiated bands from the DGGE gels was determined based on GenBank data. In total, 41.2 % (42/102) of the patients showed bacterial infection in their bile samples. This infection was detected in 21 % (4/19), 45.4 % (5/11), 53.5 % (15/28), and 54.5 % (24/44) of patients with common bile duct stone, microlithiasis, malignancy, and gallbladder stone, respectively. Escherichia coli showed a significant association with gallstones. Polymicrobial infection was detected in 48 % of the patients. While results of the culture method established coexistence of biofilm-forming bacteria (Pseudomonas aeruginosa, E. coli, Klebsiella pneumoniae, Enterococcus spp., and Acinetobacter spp.) in different combinations, the presence of Capnocytophaga spp., Lactococcus spp., Bacillus spp., Staphylococcus haemolyticus, Enterobacter or Citrobacter spp., Morganella spp., Salmonella spp., and Helicobacter pylori was also characterized in these samples by the PCR-DGGE method. Multidrug resistance phenotypes (87.5 %) and resistance to third- and fourth-generation cephalosporins and quinolones were common in these strains, which could evolve through their selection by bile components. Ability for biofilm formation seems to be a need for polymicrobial infection in this organ.

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Acknowledgments

The study was supported by a research grant (number RIGLD 564) from the Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran. The authors appreciate all their colleagues in the Foodborne and Waterborne Diseases Research Center and the ERCP unit at Ayatollah Taleghani Hospital.

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Correspondence to M. Alebouyeh.

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Funding

This study was supported by the Shahid Beheshti University of Medical Sciences, Tehran, Iran, with grant number RIGLD 564.

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The authors have no conflict of interest to declare.

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This study was approved by the ethics committee of Shahid Beheshti University of Medical Sciences, Tehran, Iran.

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Informed consent was obtained from all individual participants for the information included in this article.

Additional information

M. Alebouyeh and A. H. M. Alizadeh contributed equally to this work.

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Tajeddin, E., Sherafat, S.J., Majidi, M.R.S. et al. Association of diverse bacterial communities in human bile samples with biliary tract disorders: a survey using culture and polymerase chain reaction-denaturing gradient gel electrophoresis methods. Eur J Clin Microbiol Infect Dis 35, 1331–1339 (2016). https://doi.org/10.1007/s10096-016-2669-x

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  • DOI: https://doi.org/10.1007/s10096-016-2669-x

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