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marA efflux pump gene expression in Salmonella enteritidis strains treated with Artemisia tournefortiana hydroalcoholic extract and comparison with commercial efflux pump inhibitor, carbonyl cyanide 3-chlorophenylhydrazone (CCCP)

Marjan Khosravani, Mohammad Mehdi Soltan Dallal, Mehdi Norouzi




Background: Salmonella enterica subsp. enterica serovar Enteritidis is a food-borne pathogenic bacterium that has recently become resistant to most quinolone antibiotics. The MarA efflux pump plays a significant role in the development of ciprofloxacin resistance in S. Enteritidis strains. The aim of this study was comparative evaluation of anti-efflux activity of Artemisia tournefortiana extract and commercial efflux inhibitor, carbonyl cyanide 3-chlorophenylhydrazone (CCCP) on marA efflux pump gene expression in S. Enteritidis clinical strains.

 Materials and Methods: In this experimental study, Artemisia tournefortiana extract was prepared using maceration method. Subsequently, MarA efflux pump was detected in 20 clinical strains of S. Enteritidis via cartwheel and PCR methods. Finally, after treatment of strains with subMIC concentration of extract and 20 µg/L and CCCP, their anti-efflux activity against MarA efflux pump was studied using Real Time PCR.

 Results: The results of cartwheel and PCR methods indicated that all of ciprofloxacin resistant strains had MarA efflux pump. Subsequently, after treatment of strains with subMIC concentration of extract and CCCP, results show that both component have the ability to inhibit the MarA efflux pump, significantly.

 Conclusion: Considering the results of MarA efflux inhibition by A. tournefortiana and CCCP, it seems that this plant can be used as a potential source of drug use as a suppository pump inhibitor instead of CCCP.



Salmonella Enteritidis, efflux pump, MarA, Artemisia tournefortiana, gene expression


- Sánchez-Vargas FM, Abu-El-Haija MA, Gómez-Duarte OG. Salmonella infections: an update on epidemiology, management and prevention. Travel Med Infect Dis. 2011;9(6):263-77.

- Gong J, Kelly P, Wang C. Prevalence and antimicrobial resistance of Salmonella enterica Serovar Indiana in China (1984-2016). Zoonoses Public Health. 2017;64(4):239-251.

- Nikaido H, Takatsuka Y. Mechanisms of RND multidrug efflux pumps. Biochim Biophys Acta. 2009;1794(5):769-81.

- Fischer N, Raunest M, Schmidt TH, Koch DC, Kandt C. Efflux pump-mediated antibiotics resistance: insights from computational structural biology. Inter Discip Sci. 2014;6(1):1-12.

- Zhang CZ, Chen PX, Yang L, Li W, Chang MX, Jiang HX. Coordinated expression of AcrAB-tolC and eight other functional efflux pumps through activating ramA and MarA in Salmonella enterica serovar Typhimurium. Microb Drug Resist. 2018; 24(2):120-125.

- Tibbetts RJ, Lin TL, Wu CC. Insertional mutation of MarA vitiates inducible multiple antimicrobial resistance in Salmonella enterica subsp.enterica serovar Choleraesuis. Vet Microbiol. 2005;109(3-4):267-74.

- Ardebili A, Talebi M, Azimi L, Rastegar Lari A. Effect of efflux pump inhibitor carbonyl cyanide 3-chlorophenylhydrazone on the minimumInhibitory concentration of ciprofloxacin in Acinetobacter baumannii clinical isolates. Jundishapur J Microbiol. 2014;7(1):e8691.

- Kayani WK, Kiani BH, Dilshad E, Mirza B. Biotechnological approaches for artemisinin production in Artemisia. World J Microbiol Biotechnol. 2018;27;34(4):54.

- Blainski A, Lopes GC, de Mello JC. Application and analysis of the folin ciocalteu method for the determination of the total phenolic content from Limonium brasiliense L. Molecules. 2013;18(6):6852-65.

- Madaan R, Bansal G, Kumar S, Sharma A. Estimation of total phenols and flavonoids in extracts of Actaea spicata roots and antioxidant activity studies. Indian J Pharm Sci. 2011;73(6):666-9.

- Sagar B, Kedare R, Singh P. Genesis and development of DPPH method of antioxidant assa y. J Food Sci Technol. 2011;48(4): 412–422

- Clinical and laboratory standards institute (CLSI), Performance standards for antimicrobial susceptibility testing; 16th informational supplement. CLSI. Wayne Pa. 2015;26(3):34-38.

- Martins M, McCusker MP, Viveiros M, Couto I, Fanning S, Pagès JM, AMarAl L. A Simple method for assessment of MDR bacteria for over-expressed efflux pumps. Open Microbiol J. 2013;22:(7):72-82.

- Furutani S, Kajiya M, Aramaki N and Kubo J. Rapid detection of Salmonella enterica in food using a compact disc-shaped device. Micromachines (Basel). 2016;7(1):10-17.

- Gislene G, Nascimento F, J Locatelli J, Paulo C, Giuliana L. Antibacterial activity of plant extract and phytochemicals on antibiotic resistance Bacterial. Braz J Microbiol. 2000; 31:(4): 247-256.

- Zhong HQ, Zhang S, Pan H, Cai T. Influence of induced ciprofloxacin resistance

on efflux pump activity of Klebsiella pneumonia. J Zhejiang Univ Sci B. 2013;14(9): 837–843.

- Haselbeck AH, Panzner U, Im J, Baker S, Meyer CG, Marks F. Current perspectives on invasive nontyphoidal Salmonella disease. Curr Opin Infect Dis. 2017;30(5):498-503.

- Mąka Ł, Popowska M. Antimicrobial resistance of Salmonella spp. isolated from food. Rocz Panstw Zakl Hig. 2016;67(4):343-358.

- Tegos GP1, Haynes M, Strouse JJ, Khan MM, Bologa CG, Oprea TI, Sklar LA. Microbial efflux pump inhibition: tactics and strategies. Curr Pharm. 2011;17(13):1291-302.

- Temraz A, El-Tantawy WH. Characterization of antioxidant activity of extract from Artemisia vulgaris. Pak J Pharm Sci. 2008; 21(4):321-6.

- Sengul M, Ercisli S, Yildiz H, Gungor N, Kavaz A, Cetin B. Antioxidant, Antimicrobial Activity and Total Phenolic Content within the Aerial Parts of Artemisia absinthum, Artemisia santonicum and Saponaria officinalis. Iran J Pharm Res. 2011;10(1):49-56.

- Stavri M, Piddock LJ, Gibbons S. Bacterial efflux pump inhibitors from natural sources. J Antimicrob Chemother. 2007;59(6):1247-60.

- Lomovskaya O, Watkins W. Inhibition of efflux pumps as a novel approach to combat drug resistance in bacteria. J Mol Microbiol Biotechnol. 2001;3(2):225-36.

- Maisuria VB, Hosseinidoust Z1, Tufenkji N. Polyphenolic extract from maple syrup potentiates antibiotic susceptibility and reduces biofilm formation of pathogenic bacteria. Appl Environ Microbiol. 2015:81(11):3782-92.

- Jyoti M, Savita J, Urmila M, Evaluation of phytochemical and synergistic interaction between plants extract and antibiotics for efflux pump inhibitory activity against Salmonella enterica serovar typhimurium strains. International Journal of Pharmacy and Pharmaceutical Sciences. 2016;(8): 217-223.

DOI: https://doi.org/10.22037/amls.v4i1.24090


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