Molecular Characterization of Salmonella typhimurium from meat samples in Multan, Pakistan

Year 2020, Volume: 1 Issue: 2, 35 - 41, 22.04.2020

Sadaf Parvez Muhammad Safdar Yasmeen Junejo Sana Zaheer

Abstract

Global meat production has increased by 20% in the past 10 years only and has tripled over in the last 4 years. Most of the systemic intestinal infections are caused by gram-negative bacteria in humans, called Salmonella typhimurium. This research aimed to identify Salmonella typhimurium at molecular level through optimized PCR. The work was done by collecting samples from five different regions of Multan, Pakistan. The bacterial cells were grown on petri dishes and subsequently on broth media. DNA was extracted following protocol specified for bacterial cells. The quality of DNA was checked by agarose gel electrophoresis in 0.8% agarose gel for 45 minutes. The PCR was optimized by amplifying the DNA with species-specific primers. Further sensitivity of PCR was done by evaluating the identified DNA of Salmonella typhimurium by diluting the DNA up to 0.01 ng. 10% of the total collected samples for the identification of Salmonella typhimurium by PCR from the raw meat samples. This study revealed that raw meat was contaminated with Salmonella typhimurium at different meat retail shops in Multan. This study may help with the management of Salmonella typhimurium in Pakistan.

Keywords

Salmonella typhimurium, Meat samples, Bacterial culture, DNA extraction, Molecular Characterization

References

• D’Aoust, J.-Y., & Maurer, J. (2007). Salmonella species Food Microbiology: Fundamentals and Frontiers, Third Edition (pp. 187-236): American Society of Microbiology.
• Dahal, N., Ellerbroek, L., & Poosaran, N. (2007). Prevalence and antimicrobial resistance of Salmonella in imported chicken carcasses in Bhutan. National Cent Anim Health, 1, 1-92.
• Dandekar, T., Fieselmann, A., Popp, J., & Hensel, M. (2012). Salmonella enterica: a surprisingly well-adapted intracellular lifestyle. Frontiers in microbiology, 3, 164.
• Eyigor, A., Carli, K., & Unal, C. (2002). Implementation of real‐time PCR to tetrathionate broth enrichment step of Salmonella detection in poultry. Letters in applied microbiology, 34(1), 37-41.
• Finiay, B., & Falkow, S. (1989). Salmonella as an intracellular parasite. Molecular microbiology, 3(12), 1833-1841.
• GUPTA, A., PATEL, S. S., LANGUTE, S. M., KOLEKAR, M. R., BHOSALE, H. P., SHINDE, S. K., . . . SHINDE, B. (2016). Bacterial diseases of livestock animals and their impact on human health. Asian J Pharm Clin Res, 6, 8-11.
• Kirk, M. D., Pires, S. M., Black, R. E., Caipo, M., Crump, J. A., Devleesschauwer, B., . . . Hald, T. (2015). World Health Organization estimates of the global and regional disease burden of 22 foodborne bacterial, protozoal, and viral diseases, 2010: a data synthesis. PLoS medicine, 12(12), e1001921.
• Maniatis, T., Fritsch, E. F., & Sambrook, J. (1982). Molecular cloning: a laboratory manual (Vol. 545): Cold spring harbor laboratory Cold Spring Harbor, NY.
• Martínez, L. C., Yakhnin, H., Camacho, M. I., Georgellis, D., Babitzke, P., Puente, J. L., & Bustamante, V. H. (2011). Integration of a complex regulatory cascade involving the SirA/BarA and Csr global regulatory systems that controls expression of the Salmonella SPI‐1 and SPI‐2 virulence regulons through HilD. Molecular Microbiology, 80(6), 1637-1656.
• Pui, C., Wong, W., Chai, L., Tunung, R., Jeyaletchumi, P., Hidayah, N., Son, R. (2011). Salmonella: A foodborne pathogen. International Food Research Journal, 18(2).
• Rahn, K., De Grandis, S., Clarke, R., McEwen, S., Galan, J., Ginocchio, C., Gyles, C. (1992). Amplification of an invA gene sequence of Salmonella typhimurium by polymerase chain reaction as a specific method of detection of Salmonella. Molecular and cellular probes, 6(4), 271-279.
• Schönenbrücher, V., Mallinson, E. T., & Bülte, M. (2008). A comparison of standard cultural methods for the detection of foodborne Salmonella species including three new chromogenic plating media. International journal of food microbiology, 123(1-2), 61-66.
• Settanni, L., & Corsetti, A. (2007). The use of multiplex PCR to detect and differentiate food-and beverage-associated microorganisms: a review. Journal of Microbiological Methods, 69(1), 1-22.
• Starnbach, M. N., Falkow, S., & Tompkins, L. S. (1989). Species-specific detection of Legionella pneumophila in water by DNA amplification and hybridization. Journal of Clinical Microbiology, 27(6), 1257-1261.
• Wang, L., Wang, R., Chen, F., Jiang, T., Wang, H., Slavik, M., Li, Y. (2017). QCM-based aptamer selection and detection of Salmonella typhimurium. Food Chemistry, 221, 776-782.