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:: Volume 21, Issue 3 (10-2019) ::
J Gorgan Univ Med Sci 2019, 21(3): 120-128 Back to browse issues page
Determination of bacteria in gallbladder of patients with cholelithiasis and evaluation of pathogenic factors of the prevalent isolated bacteria
Parastoo Ehsani * 1, Fateme Farahany2 , Negar Daeizadeh3 , Melika Amya2 , Moein Saleh4 , Mina Ebrahimi-Rad5 , Karamollah Toolabi6
1- Ph.D in Biological Products, Associate Professor, Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran , p_ehsani@yahoo.com
2- Microbiologist, Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran
3- M.Sc in Biotechnoloy, Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran
4- M.Sc in Microbial Biotechnology, Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran
5- Ph.D in Medical Biotechnology, Assistant Professor , Department of Biochemistry, Pasteur Institute of Iran, Tehran, Iran
6- Associate Professor, Department of Laparoscopic Surgery, Imam Khomeini Hospital, Tehran, Iran
Abstract:   (12288 Views)
Background and Objective: Various infections and the formation of stones could be the cause for bile duct obstruction of that sterile organ. Determination of pathogenesis factors and the bacteria involved in infections are important in the prevention of disease, and cares needed following surgery. This study was done to determine the bacteria in the gallbladder of patients with cholelithiasis and evaluation of pathogenic factors of the prevalent isolated bacteria.
Methods: In this descriptive laboratory study, 35 samples of gallbladder tissues which contained gallstones were collected under sterile conditions in “Department of General Surgery of Imam Khomeini Hospital”, Tehran, Iran during 2016. The stone types were analyzed and decomposed by chemical procedures, and the bacteria existed in the tissues were also identified using biochemical experiments. The tissues with negative results in microbiological studies were looked for any contaminating bacteria, applying the DNA extracted from gallbladder tissue as a template using F27 and R1492 as the primers for PCR (Polymerase Chain Reaction) amplification of 16SrRNA gene. Those with the positive results of microbiological tests were subjected to the DNA sequencing following gel purification and blasted against the NCBI gene database. The most frequently isolated bacteria were studied according to the intensity of biofilm formation, using the microtitre plate method. CsgF and Ag43 (Flu), the genes involved in the induction of such phenotype were also analyzed in this study. The antibiotic resistance assay of the isolates was performed using disc diffusion procedure.
Results: Thirteen out of thirty five samples of post-surgery gallbladder tissues were found to be infected by different bacteria, including: Klebsiella (3 cases), Escherichia coli (4 cases), Enterobacter (1 case), Staphylococcus aureus (2 cases), Enterococci (2 cases), and Streptococcus (1 case). In 23 out of 35 samples (65.7%), no bacteria could be isolated using microbiological methods. However, in seven out of 23 samples, the amplified 16SrRNA had an indication of Klebsiella (6 cases) and Enterococcus (1 case) isolates. Therefore, the most prevalent genus in gallbladder infections was Klebsiella (47.36%). Chemical analysis showed that the highly frequent compound of gallstones (98%) were of cholesterol and bilirubin. Escherichia coli with fourcases were the highest culture growing isolated bacteria, in all of which, the biofilm formation genes were present. In the two out of four Escherichia coli isolates the intensity of biofilm formation was high. Although, in the remaining two isolates was medium. While, they were found to be sensitive to the most of the antibiotics, they showed resistance to Tetracycline, Ciprofloxacin and Ceftazidime in different ranges.
Conclusion: The present study provided evidence that non-cultural bacteria are hihly present in gallbladder infections. The high potential of the commonly isolated bacteria in biofilm formation should be taken as a warning to follow the precise protocol of antibiotic prescription for treatment of gallbladder infections.
Keywords: Choelitiasis, Entrobacteriace, Biofilm, Polymerase Chain Reaction
Full-Text [PDF 323 kb]   (13788 Downloads)    
Type of Study: Original Articles | Subject: Microbiology
References
1. Sifri CD, Madoff LC. Infections of the liver and biliary system. In: Mandell GL, Bennett JE, Dolin R. Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases. 7th ed. Philadelphia: Churchill Livingstone. 2010; pp: 1035-45.
2. Melzer M, Toner R, Lacey S, Bettany E, Rait G. Biliary tract infection and bacteraemia: presentation, structural abnormalities, causative organisms and clinical outcomes. Postgrad Med J. 2007 Dec; 83(986): 773-76. doi: 10.1136/pgmj.2007.064683
3. Namias N, Demoya M, Sleeman D, Reever CM, Raskin JB, Ginzburg E, et al. Risk of postoperative infection in patients with bactibilia undergoing surgery for obstructive jaundice. Surg Infect (Larchmt). 2005; 6(3): 323-28. doi: 10.1089/sur.2005.6.323
4. Biscione FM, Couto RC, Pedrosa TM, Neto MC. Comparison of the risk of surgical site infection after laparoscopic cholecystectomy and open cholecystectomy. Infect Control Hosp Epidemiol. 2007 Sep; 28(9): 1103-6. doi: 10.1086/519931
5. Vollmer CM Jr, Callery MP. Biliary injury following laparoscopic cholecystectomy: why still a problem? Gastroenterology. 2007 Sep; 133(3): 1039-41. doi: 10.1053/j.gastro.2007.07.041
6. Vafaeimanesh J, Alebouyeh M, Seyyedmajidi M, Tajeddin E, Sherafat SJ, et al. Molecular Detection of Helicobacter Species and Other Bacteria in Human Bile Samples of Patients with Biliary Diseases. J Gastrointest Dig Syst. 2012; 2: 106. doi:10.4172/2161-069X.1000106
7. Vaishnavi C, Singh S, Kochhar R, Bhasin D, Singh G, Singh K. Prevalence of Salmonella enterica serovar typhi in bile and stool of patients with biliary diseases and those requiring biliary drainage for other purposes. Jpn J Infect Dis. 2005 Dec; 58(6): 363-65.
8. Manolis EN, Filippou DK, Papadopoulos VP, Kaklamanos I, Katostaras T, Christianakis E, et al. The culture site of the gallbladder affects recovery of bacteria in symptomatic cholelithiasis. J Gastrointestin Liver Dis. 2008 Jun; 17(2): 179-82.
9. Austin JW, Sanders G, Kay WW, Collinson SK. Thin aggregative fimbriae enhance Salmonella enteritidis biofilm formation. FEMS Microbiol Lett. 1998 May; 162(2): 295-301. doi: 10.1111/j.1574-6968.1998.tb13012.x
10. Kikuchi T, Mizunoe Y, Takade A, Naito S, Yoshida S. Curli fibers are required for development of biofilm architecture in Escherichia coli K-12 and enhance bacterial adherence to human uroepithelial cells. Microbiol Immunol. 2005; 49(9): 875-84.
11. Van Houdt R, Michiels CW. Role of bacterial cell surface structures in Escherichia coli biofilm formation. Res Microbiol. 2005 Jun-Jul; 156(5-6): 626-33. doi: 10.1016/j.resmic.2005.02.005
12. Vidal O, Longin R, Prigent-Combaret C, Dorel C, Hooreman M, Lejeune P. Isolation of an Escherichia coli K-12 mutant strain able to form biofilms on inert surfaces: involvement of a new ompR allele that increases curli expression. J Bacteriol. 1998 May; 180(9): 2442-49.
13. Hammar M, Arnqvist A, Bian Z, Olsén A, Normark S. Expression of two csg operons is required for production of fibronectin- and congo red-binding curli polymers in Escherichia coli K-12. Mol Microbiol. 1995 Nov; 18(4): 661-70.
14. Robinson LS, Ashman EM, Hultgren SJ, Chapman MR. Secretion of curli fibre subunits is mediated by the outer membrane-localized CsgG protein. Mol Microbiol. 2006 Feb; 59(3): 870-81. doi: 10.1111/j.1365-2958.2005.04997.x
15. Barnhart MM, Chapman MR. Curli biogenesis and function. Annu Rev Microbiol. 2006; 60: 131-47. doi: 10.1146/annurev.micro.60.080805.142106
16. Chapman MR, Robinson LS, Pinkner JS, Roth R, Heuser J, Hammar M, et al. Role of Escherichia coli curli operons in directing amyloid fiber formation. Science. 2002 Feb; 295(5556): 851-55. doi: 10.1126/science.1067484
17. Hammar M, Bian Z, Normark S. Nucleator-dependent intercellular assembly of adhesive curli organelles in Escherichia coli. Proc Natl Acad Sci U S A. 1996 Jun; 93(13): 6562-66. doi: 10.1073/pnas.93.13.6562
18. Loferer H, Hammar M, Normark S. Availability of the fibre subunit CsgA and the nucleator protein CsgB during assembly of fibronectin-binding curli is limited by the intracellular concentration of the novel lipoprotein CsgG. Mol Microbiol. 1997 Oct; 26(1): 11-23.
19. Sharma G, Sharma S, Sharma P, Chandola D, Dang S, Gupta S, et al. Escherichia coli biofilm: development and therapeutic strategies. J Appl Microbiol. 2016 Aug; 121(2): 309-19. doi: 10.1111/jam.13078
20. Reisner A, Haagensen JA, Schembri MA, Zechner EL, Molin S. Development and maturation of Escherichia coli K-12 biofilms. Mol Microbiol. 2003 May; 48(4): 933-46.
21. van der Woude MW, Henderson IR. Regulation and function of Ag43 (flu). Annu Rev Microbiol. 2008; 62: 153-69. doi: 10.1146/annurev.micro.62.081307.162938
22. Massarrat S. Prevalence of gallstone disease in Iran. J Gastroenterol Hepatol. 2001 May; 16(5): 564-67.
23. Weisburg WG, Barns SM, Pelletier DA, Lane DJ. 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol. 1991 Jan; 173(2): 697-703. doi: 10.1128/jb.173.2.697-703.1991
24. O'Toole GA. Microtiter dish biofilm formation assay. J Vis Exp. 2011 Jan; (47) pii: 2437. doi: 10.3791/2437
25. Reshetnikov OV, Ryabikov AN, Shakhmatov SG, Malyutina SK. Gallstone disease prevalence in Western Siberia: cross-sectional ultrasound study versus autopsy. J Gastroenterol Hepatol. 2002 Jun; 17(6): 702-707.
26. Ersumo T. Gallstone disease in a teaching hospital, Addis Ababa: a 5-year review. Ethiop Med J. 2006 Jan; 44(1): 49-59.
27. Stewart L, Griffiss JM, Jarvis GA, Way LW. Gallstones containing bacteria are biofilms: bacterial slime production and ability to form pigment solids determines infection severity and bacteremia. J Gastrointest Surg. 2007 Aug; 11(8): 977-83. doi: 10.1007/s11605-007-0168-1
28. Swidsinski A, Schlien P, Pernthaler A, Gottschalk U, Bärlehner E, Decker G, et al. Bacterial biofilm within diseased pancreatic and biliary tracts. Gut. 2005 Mar; 54(3): 388-95. doi: 10.1136/gut.2004.043059
29. Shen H, Ye F, Xie L, Yang J, Li Zh, Xu P, et al. Metagenomic sequencing of bile from gallstone patients to identify different microbial community patterns and novel biliary bacteria. Sci Rep. 2015; 5: 17450. doi: 10.1038/srep17450
30. Tajeddin E, Jahani Sherafat S, Seyyed Majidi MR, Alebouyeh M, Nazemalhosseini Mojarad E, Pourhossengholi A, et al. [The Frequency of Bacterial Agents in the Bile Juice of Patients with Bile Stones and Biliary Disorders]. Med Lab J. 2011; 5(2): 34-43. [Article in Persian]
31. Tajbakhsh E, Ahmadi P, Abedpour-Dehkordi E, Arbab-Soleimani N, Khamesipour F. Biofilm formation, antimicrobial susceptibility, serogroups and virulence genes of uropathogenic E. coli isolated from clinical samples in Iran. Antimicrobial Resistance & Infection Control. 2016; 5: 11. https://doi.org/10.1186/s13756-016-0109-4
32. Taghadosi R, Shakibaie MR, Ghanbarpour R, Hosseini-Nave H. Role of antigen-43 on biofilm formation and horizontal antibiotic resistance gene transfer in non-O157 Shiga toxin producing Escherichia coli strains. Iran J Microbil. 2017; 9(2): 98-96.
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Ehsani P, Farahany F, Daeizadeh N, Amya M, Saleh M, Ebrahimi-Rad M et al . Determination of bacteria in gallbladder of patients with cholelithiasis and evaluation of pathogenic factors of the prevalent isolated bacteria. J Gorgan Univ Med Sci 2019; 21 (3) :120-128
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Volume 21, Issue 3 (10-2019) Back to browse issues page
مجله دانشگاه علوم پزشکی گرگان Journal of Gorgan University of Medical Sciences
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