Evaluating the virulence factors of Salmonella typhi isolated from children suffering from severe diarrhea using molecular genetic techniques

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DOI https://doi.org/10.13057/biodiv/d240653
Issue
Vol. 24 No. 6 (2023)
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Articles
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This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

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AMANY SHAKEIR JABER ♥
Department of Pathological Analysis, College of Sciences, University of Thi-Qar. Nasiriyah, Iraq

Abstract

Abstract. Jaber AS. 2023. Evaluating the virulence factors of Salmonella typhi isolated from children suffering from severe diarrhea using molecular genetic techniques. Biodiversitas 24: 3543-3549. Salmonella enterica subsp. serotype Typhi is the bacteria that causes typhoid fever. Although its prevalence has significantly decreased in affluent nations, it continues to be a leading cause of illness and mortality in emerging nations. Because of changes in ecology, eating habits, and agricultural and food production techniques, Salmonella infections have increased in frequency in industrialized countries. This study aimed to use qualitative RT-PCR (based on the ttr gene) and conventional PCR to measure the frequency of Salmonella typhi in Iraqi children with chronic diarrhea (based on PagN and TolC genes). This study included 100 stool samples collected from children suffering from continuous diarrhea. Those samples were inoculated on CHROM-agar and incubated 37°C for 18-24 hours, then Salmonella was identified using a Vitek 2 compact system. DNA has been extracted, and qRT-PCR has been done to detect Salmonella sp. by the amplification of the ttr gene. Then PCR also done to amplify TolC and PagN genes. The amplified products then sequenced to reveal mutations within the amplified regions. The results of PagN gene showed 41 samples were positive, and 40 samples were positive for TolC. Results of qRT-PCR according to the ttr gene amplification showed 38 samples positive. The blood group O+ showed a higher number of positive samples (13), and males showed higher positive samples than females (23 and 15, respectively). Sequencing revealed various number of mutations within the amplified regions of both PagN and TolC in the samples which isolated from children with the longest period of illness.

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References
Al-Zubaidy, K. I. (2020). Bacterial Vulnerability and Blood Types, Salmonella and Escherichia coli as A Study Case. Journal of Basrah Researches ((Sciences)), 46(2).
Belden, W. J., & Miller, S. I. (1994). Further characterization of the PhoP regulon: identification of new PhoP-activated virulence loci. Infection and immunity, 62(11), 5095-5101. DOI: 10.1128/IAI.62.11.5095-5101.1994.
Chirambo, A. C., Nyirenda, T. S., Jambo, N., Msefula, C., Kamng'ona, A., Molina, S., ... & Gordon, M. A. (2020). Performance of molecular methods for the detection of Salmonella in human stool specimens. Wellcome open research, 5. DOI: 10.12688/WELLCOMEOPENRES.16305.2.
Chowdhury, N., Suhani, S., Purkaystha, A., Begum, M. K., Raihan, T., Alam, M. J., ... & Azad, A. K. (2019). Identification of AcrAB-TolC efflux pump genes and detection of mutation in efflux repressor AcrR from omeprazole responsive multidrug-resistant Escherichia coli isolates causing urinary tract infections. Microbiology insights, 12, 1178636119889629. DOI: 10.1177/1178636119889629.
Dashtbani-Roozbehani, A., & Brown, M. H. (2021). Efflux pump mediated antimicrobial resistance by Staphylococci in health-related environments: Challenges and the quest for inhibition. Antibiotics, 10(12), 1502. DOI: 10.3390/ANTIBIOTICS10121502.
Diarrhoea - UNICEF DATA.” https://data.unicef.org/topic/child-health/diarrhoeal-disease/ (accessed Sep. 25, 2022).
Dró?d?, M., Ma?aszczuk, M., Paluch, E., & Pawlak, A. (2021). Zoonotic potential and prevalence of Salmonella serovars isolated from pets. Infection Ecology & Epidemiology, 11(1), 1975530. DOI: 10.1080/20008686.2021.1975530.
Dró?d?, M., Ma?aszczuk, M., Paluch, E., & Pawlak, A. (2021). Zoonotic potential and prevalence of Salmonella serovars isolated from pets. Infection Ecology & Epidemiology, 11(1), 1975530. DOI: 10.1080/20008686.2021.1975530.
Folkesson, A., Advani, A., Sukupolvi, S., Pfeifer, J. D., Normark, S., & Löfdahl, S. (1999). Multiple insertions of fimbrial operons correlate with the evolution of Salmonella serovars responsible for human disease. Molecular microbiology, 33(3), 612-622. DOI: 10.1046/j.1365-2958.1999.01508.x.
Harb, A., O’Dea, M., Abraham, S., & Habib, I. (2019). Childhood diarrhoea in the Eastern Mediterranean region with special emphasis on non-typhoidal Salmonella at the human–food interface. Pathogens, 8(2), 60. DOI: 10.3390/PATHOGENS8020060.
Kasturi, K. N., & Drgon, T. (2017). Real-time PCR method for detection of Salmonella spp. in environmental samples. Applied and environmental microbiology, 83(14), e00644-17. DOI: 10 .1128/AEM.00644-17.
Kolahi, A. A., Nabavi, M., & Sohrabi, M. R. (2008). Epidemiology of acute diarrheal diseases among children under 5 years of age in Tehran, Iran. Archives of Clinical Infectious Diseases, 3(4), 193-8.
Lambert, M. A., & Smith, S. G. (2008). The PagN protein of Salmonella enterica serovar Typhimurium is an adhesin and invasin. BMC microbiology, 8(1), 1-11. DOI: 10.1186/1471-2180-8-142.
Lambert, M. A., & Smith, S. G. (2009). The PagN protein mediates invasion via interaction with proteoglycan. FEMS microbiology letters, 297(2), 209-216.DOI: 10.1111/j.1574-6968.2009.01666.x.
Liu, D., Wang, Y., Wang, X., Ou, D., Ling, N., Zhang, J., ... & Ye, Y. (2021). Role of the multiple efflux pump protein TolC on growth, morphology, and biofilm formation under nitric oxide stress in Cronobacter malonaticus. JDS Communications, 2(3), 98-103. DOI: 10.3168/jdsc.2020-0040.
Lowden, P., Wallis, C., Gee, N., & Hilton, A. (2015). Investigating the prevalence of Salmonella in dogs within the Midlands region of the United Kingdom. BMC Veterinary Research, 11(1), 1-6. DOI: 10.1186/S12917-015-0553-Z.
Mokdad, A. H., Khalil, I., BCHERAOUI, C. E., Charara, R., Moradi-Lakeh, M., Afshin, A., ... & Murray, C. J. (2018). Burden of diarrhea in the Eastern Mediterranean Region, 1990-2015: Findings from the Global Burden of Disease 2015 study. International Journal of Public Health. DOI: 10.1007/S00038-017-1008-Z.
Nair, S., Patel, V., Hickey, T., Maguire, C., Greig, D. R., Lee, W., ... & Chattaway, M. A. (2019). Real-time PCR assay for differentiation of typhoidal and nontyphoidal Salmonella. Journal of clinical microbiology, 57(8), e00167-19. DOI: 10.1128/JCM.00167-19.
Porras, A. M., Shi, Q., Zhou, H., Callahan, R., Montenegro-Bethancourt, G., Solomons, N., & Brito, I. L. (2021). Geographic differences in gut microbiota composition impact susceptibility to enteric infection. Cell Reports, 36(4), 109457. DOI: 10.1016/J.CELREP.2021.109457.
Raspoet, R., Eeckhaut, V., Vermeulen, K., De Smet, L., Wen, Y., Nishino, K., ... & Van Immerseel, F. (2019). The Salmonella Enteritidis TolC outer membrane channel is essential for egg white survival. Poultry science, 98(5), 2281-2289. DOI: 10.3382/ps/pey584.
Rooks, M. G., & Garrett, W. S. (2016). Gut microbiota, metabolites and host immunity. Nature reviews immunology, 16(6), 341-352. DOI: 10.1038/NRI.2016.42.
Ross, A. A., Rodrigues Hoffmann, A., & Neufeld, J. D. (2019). The skin microbiome of vertebrates. Microbiome, 7(1), 1-14. DOI: 10.1186/S40168-019-0694-6.
Siala, M., Barbana, A., Smaoui, S., Hachicha, S., Marouane, C., Kammoun, S., ... & Messadi-Akrout, F. (2017). Screening and detecting Salmonella in different food matrices in Southern Tunisia using a combined enrichment/real-time PCR method: Correlation with conventional culture method. Frontiers in microbiology, 8, 2416. DOI: 10.3389/FMICB.2017.02416/BIBTEX.
Siziya, S., Muula, A. S., & Rudatsikira, E. (2009). Diarrhoea and acute respiratory infections prevalence and risk factors among under-five children in Iraq in 2000. Italian journal of pediatrics, 35(1), 1-9. DOI: 10.1186/1824-7288-35-8.
Wotzka, S. Y., Nguyen, B. D., & Hardt, W. D. (2017). Salmonella Typhimurium diarrhea reveals basic principles of enteropathogen infection and disease-promoted DNA exchange. Cell host & microbe, 21(4), 443-454. DOI: 10.1016/J.CHOM.2017.03.009.
Yue, M., Li, X., Liu, D., & Hu, X. (2020). Serotypes, antibiotic resistance, and virulence genes of Salmonella in children with diarrhea. Journal of Clinical Laboratory Analysis, 34(12), e23525. DOI: 10.1002/JCLA.23525.
Zgurskaya, H. I., Krishnamoorthy, G., Ntreh, A., & Lu, S. (2011). Mechanism and function of the outer membrane channel TolC in multidrug resistance and physiology of enterobacteria. Frontiers in microbiology, 2, 189. DOI: 10.3389/fmicb.2011.00189.