Molecular identification of CdtB and TviA virulence genes of Salmonella typhi isolated from cholelithiasis patients in Erbil City, Iraq
Article Main
Abstract
Gallstone disease (GSD), particularly cholecystitis accompanied by gallstones, is a prevalent global health concern with significant morbidity and mortality rates. Typhoid fever is a public health issue in low- and middle-income countries. The severity of the pathogenesis depends on Salmonella’s possession of several virulence factors encoded on Salmonella pathogenicity islands (SPIs). This study aimed to isolate and identify Salmonella typhi, the causative agent of typhoid fever, in clinical samples from cholelithiasis patients in Erbil City, Iraq. A cross-sectional study was conducted from October 2023 to March 2024, involving 125 patients diagnosed with gallbladder disease. Gallstone, bile, and gallbladder tissue samples were collected and analyzed for bacterial growth. Biochemical and cultural studies confirmed antibiotic susceptibility and Vitek2 system identification. The present study used a traditional PCR assay to detect Salmonella typhi pathogenicity genes. This study found the virulence genes CtdB and TviA using specific primers. Out of 125 samples, 101 (80.8%) showed bacterial growth, with 8 (6.4%) positive for S. typhi and 93 (74.4%) positive for other bacterial species. Gallstones had the highest proportion of S. typhi isolation 5 (4%). Antibiotic susceptibility testing revealed significant resistance to commonly used antibiotics like ampicillin, ciprofloxacin, chloramphenicol, and trimethoprim-sulfamethoxazole. However, the strains demonstrated better susceptibility to various antibiotics. The molecular result showed the virulence genes, including CtdB and TviA, were detected in all (100%) isolated strains. S. typhi's virulence characteristics significantly contributed to gallbladder infections and multidrug-resistant strains (MDR).
Article Details
Article Details
Keywords
Salmonella typhi, Cholelithiasis, Polymerase chain reaction (PCR), Virulence genes
References
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Sadeq Noomi, B. & Ayad Majeed, K. (2018). Investigation of Carrier Persons of Salmonella Typhi in Cholelithias is Patients in Kirkuk Province. Kirkuk University Journal-Scientific Studies, 13(2), 40-48. http://dx.doi.org/10.32894/kujss.2018.145713.
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Al-Sanjary, S. I. & Al-Rawi, A. M. M. (2022). The Correlation Between Salmonella typhi Associated Gallstone Formation and Gallbladder Cancer. Iraqi Journal of Science, 3332-3339. https://doi.org/10.24996/ijs.2022.63.8.8.
Al-Turaihi, T. S. A., Mubark, H. A., Hadi, Z. J., Akool, M. A. & Al-Nafak, R. T. (2023). ANTIBIOTIC RESISTANCE OF SALMONELLA TYPHI CARRIER ASSOCIATED WITH GALL BLADDER CHRONIC INFECTION IN AL-NAJAF PROVINCE. INDEXED IN PUBMED/MEDLINE, SCOPUS, EMBASE, EBSCO, INDEX COPERNICUS, POLISH MINISTRY OF EDUCATION AND SCIENCE, POLISH MEDICAL BIBLIOGRAPHY, 76(1), 46-51. https://doi.org/10.36740/wlek202301106.
Al-Zuharri, O. A. R. (2011). Isolation and identification of bacteria from patients with cholecystits and cholelithiasis undergoing cholecystectomy. Al-Kufa J Biol, 3. https://journal.uokufa.edu.iq/index.php/ajb/article/view/9345.
Atlaw, N. A., Keelara, S., Correa, M., Foster, D., Gebreyes, W., Aidara-Kane, A. & Fedorka-Cray, P. J. (2022). Evidence of sheep and abattoir environment as important reservoirs of multidrug resistant Salmonella and extended-spectrum beta-lactamase Escherichia coli. International Journal of Food Microbiology, 363, 109516. https://doi.org/10.1016/j.ijfoodmicro.2021.109516.
Capoor, M. R., Nair, D., Khanna, G., Krishna, S. V., Chintamani, M. S. & Aggarwal, P. (2008). Microflora of bile aspirates in patients with acute cholecystitis with or without cholelithiasis: a tropical experience. Brazilian Journal of Infectious Diseases, 12, 222-225. https://doi.org/10.1590/s1413-86702008000300012.
Cetta, F. M. (1986). Bile infection documented as initial event in the pathogenesis of brown pigment biliary stones. Hepatology, 6(3), 482-489. https://doi.org/10.1002/hep.1840060327.
Chia, T. W. R., Goulter, R. M., McMeekin, T., Dykes, G. A. & Fegan, N. (2009). Attachment of different Salmonella serovars to materials commonly used in a poultry processing plant. Food microbiology, 26(8), 853-859. https://doi.org/10.1016/j.fm.2009.05.012.
Di Domenico, E. G., Cavallo, I., Pontone, M., Toma, L. & Ensoli, F. (2017). Biofilm producing Salmonella typhi: chronic colonization and development of gallbladder cancer. International journal of molecular sciences, 18(9), 1887. https://doi.org/10.3390%2Fijms18091887.
Dolecek, C., Phi La, T. T., Rang, N. N., Phuong, L. T., Vinh, H., Tuan, P. Q. & Farrar, J. (2008). A multi-center randomised controlled trial of gatifloxacin versus azithromycin for the treatment of uncomplicated typhoid fever in children and adults in Vietnam. PLoS One, 3(5), e2188. https://doi.org/10.1371/journal.pone.0002188.
El Morabet, R., Khan, R. A., Alsubih, M., Khan, N. A., Yusuf, M., Khan, P.& Lutsak, O. (2023). Epidemiology study of Diarrhoea, Cholera, Typhoid, Hepatitis A and Hepatitis E in Middle East and North Africa Region. Ecological Questions, 34(4), 1-21. https://doi.org/10.12775/EQ.2023.044.
Gilmore, B. F., & Denyer, S. P. (Eds.). (2023). Hugo and Russell's pharmaceutical microbiology. John Wiley & Sons. https://onlinelibrary.wiley.com/doi/book/10.1002/97 80470988329#:~:text=DOI%3A10.1002/9780470988329.
González, J. F., Hitt, R., Laipply, B. & Gunn, J. S. (2022). The effect of the gallbladder environment during chronic infection on Salmonella persister cell formation. Microorganisms, 10(11), 2276. https://doi.org/10.3390/microorganisms10112276.
Gonzalez-Escobedo, G. & Gunn, J. S. (2013). Identification of Salmonella enterica serovar Typhimurium genes regulated during biofilm formation on cholesterol gallstone surfaces. Infection and immunity, 81(10), 3770-3780. https://doi.org/10.1128/iai.00647-13.
Hanan, Z. K., Mezal, E. H. & Saleh, M. B. (2021). Molecular Detection of Quinolones Resistance Gens of Salmonella Typhi from Gallbladder of Patients Undergoing to Cholecystectomy in Thi-Qar province/Iraq. Annals of the Romanian Society for Cell Biology, 25(6), 268-280. http://annalsofrscb.ro/index.php/journal/article/view/5278.
Hassena, A. B., Haendiges, J., Zormati, S., Guermazi, S., Gdoura, R., Gonzalez-Escalona, N. & Siala, M. (2021). Virulence and resistance genes profiles and clonal relationships of non-typhoidal food-borne Salmonella strains isolated in Tunisia by whole genome sequencing. International Journal of Food Microbiology, 337, 108941. https://doi.org/10.1016/j.ijfoodmicro.2020.108941.
Hasson, S. O. & Abady, N. (2019). Comparative studies between using antimicrobial effect with silver nano particles effects for E. coli. Plant Archives, 19(2), 1872-1876. https://www.researchgate.net/publication/337567176.
Hawar, T. S., Shareef, H. A. & Mahdi, N. B. (2019). ISOLATION AND IDENTIFICATION OF AEROBIC BACTERIA FROM DIFFERENT CULTURE SITE OF GALLBLADDER IN PATIENTS WITH CHOLELITHIASIS. Biochemical and Cellular Archives, 19(1), 281-285. http://www.connectjournals.com/toc2.php?abstract=2948601 H_281A.pdf&&bookmark=CJ-033216& &issue_id=01&&yaer=2019.
Johnson, R., Mylona, E. & Frankel, G. (2018). Typhoidal Salmonella: Distinctive virulence factors and pathogenesis. Cellular microbiology, 20(9), e12939. https://doi.org/10.1111/cmi.12939.
K Hanan, Z., B Saleh, M., H Mezal, E., S Issa, M. A., Q Al-jauher, R. & A Akmoush, M. (2021). Phylogenitic analysis of Biofilm Association Protein (BapA) amplicons in Salmonella Typhi Carrier in Gallbladder Diseases Patients in Thi-Qar Province/Iraq. Int. J. of Aquatic Science, 12(2), 931-939. https://www.researchgate.net/publication/381829815
Kerim, U. A., Kareem, A. A. & Saleem, A. J. (2023). Molecular identification of some Salmonella Typhi virulence genes from individuals with typhoid in Baghdad. History of Medicine, 9(1), 1175-1181. http://dx.doi.org/10.17720/2409-5834.v9.1.2023.139.
Koshiol, J., Wozniak, A., Cook, P., Adaniel, C., Acevedo, J., Azócar, L. & Hildesheim, A. (2016). Salmonella enterica serovar Typhi and gallbladder cancer: a case–control study and meta‐analysis. Cancer Medicine, 5(11), 3310-3235. https://doi.org/10.1002/cam4.915.
Lee, K. M., Runyon, M., Herrman, T. J., Phillips, R. & Hsieh, J. (2015). Review of Salmonella detection and identification methods: Aspects of rapid emergency response and food safety. Food control, 47, 264-276. https://doi.org/10.1016/j.foodcont.2014.07.011.
Liaquat, S., Sarwar, Y., Ali, A., Haque, A., Farooq, M., Martinez-Ballesteros, I. & Bikandi, J. (2018). Virulotyping of Salmonella enterica serovar Typhi isolates from Pakistan: Absence of complete SPI-10 in Vi negative isolates. PLoS Neglected Tropical Diseases, 12(11), e0006839. https://doi.org/10.1371%2Fjournal.pntd.0006839.
Mansour, M. A., Zaher, T., Ibrahim, A., Ahmed, A., Ibrahim, I., Goda, A. & Ahmed, N. (2012). Role of gallstones in typhoid carriage in Egyptian patients. Journal of Microbiology and Infectious Diseases, 2(04), 142-149. http://dx.doi.org/10.5799/jmid.123127.
Muralidhar, S. (2019). Lippincott Illustrated Reviews Microbiology. Wolters Kluwer India Pvt Ltd. https://doi.org/10.4103/0377-4929.313309.
Parry, C. M., Thieu, N. T. V., Dolecek, C., Karkey, A., Gupta, R., Turner, P. & Baker, S. (2015). Clinically and microbiologically derived azithromycin susceptibility breakpoints for Salmonella enterica serovars Typhi and Paratyphi A. Antimicrobial agents and chemotherapy, 59(5), 2756-2764. https://doi.org/10.1128/aac.04729-14.
Radi, N. A. A. M., Fahmy, A., Singer, T. H. & Abdel-Aziz, M. A. B. T. (2023). The Correlation between Ciprofloxacin Resistant Salmonella Strains and Its Ability to Biofilm Formation. Egyptian Journal of Medical Microbiology, 32(3), 71-76. https://doi.org/10.21608/ejmm.2023.305231.
Rajab, N. A. & Turki, A. M. (2021). Assessment of Histopathological Changes in the Liver and Spleen of Mice Infected with Salmonella typhi Following Treatment with Ciprofloxacin. Iraqi Journal of Science, 761-768. https://doi.org/10.24996/ijs.2021.62.3.6.
Rehman, F., Tareen, A. M., Taj, M. K. & Khan, S. U. (2021). A Comprehensive Review on Salmonella Typhi: Pathogenesis, Clinical Features and Antibiotic Resistance Patterns. Pak-Euro Journal of Medical and Life Sciences, 4(Special Is), S172-S180. https://doi.org/10.31580/pjmls.v4iSpecial%20Is.1549.
Sadeq, T. N. & Rasha, K. J. (2017). Knowledge, attitude and practice of mothers towards typhoid fever disease. Iraqi JMS. 2017; Vol. 15 (1). 71-77. https://doi.org/10.22578/ijms.15.1.9.
Sadeq Noomi, B. & Ayad Majeed, K. (2018). Investigation of Carrier Persons of Salmonella Typhi in Cholelithias is Patients in Kirkuk Province. Kirkuk University Journal-Scientific Studies, 13(2), 40-48. http://dx.doi.org/10.32894/kujss.2018.145713.
Sahu, S., Karicheri, R. & Chauhan, S. P. (2021). Phenotypic and genotypic characterization of multidrug resistant Salmonella enterica serovar Typhi isolated from a tertiary care center. European Journal of Molecular and Clinical Medicine, 8(4), 1303-1312. https://link.gale.com/apps/doc/A698308214/AONE?u=anon~6f197a80&sid=googleScholar&xid=880df77a.
Saleh, M. B., Mezal, H. E. & Hanan, K. Z. (2016). Antimicrobial resistance, Virulence profiles of Salmonella enterica serovar Typhimurium isolated from diarrheal children in Thi-Qar province during 2015. University of Thi-Qar Journal of Science, 6(1), 3-8. https://doi.org/10.32792/utq/utjsci/v6i1.47.
Santander, J. M., Roland, K. L. & Curtiss III, R. (2008). Regulation of Vi capsular polysaccharide synthesis in Salmonella enterica serotype Typhi. Journal of infection in developing countries, 2(6), 412. https://doi.org/10.3855/jidc.154.
Shareef, H.A. (2005). Study on virulence factors of Salmonella isolates from patients in Kirkuk city and compared with some standard isolates", PhD thesis, Biology department, Science collage, Mosul university, Iraq.
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Molecular identification of CdtB and TviA virulence genes of Salmonella typhi isolated from cholelithiasis patients in Erbil City, Iraq. (2024). Journal of Applied and Natural Science, 16(4), 1476-1484. https://doi.org/10.31018/jans.v16i4.5922
