Abstract
Salmonella is one among the most versatile and resilient enteric pathogens that is known to have developed various survival strategies within the host system. The ability of the bacteria to circumvent the physiological parameters as well as dodge the antimicrobial stress environment within the host is one of the most crucial steps in establishing an infection. With an alarming rise in multi-drug resistant serovars of non-typhoidal Salmonella and lack of vaccine for combatting the infections, behaviour of the bacteria in the presence of host physiological conditions (NaCl, high and low iron) and antibiotics will help in understanding the survival strategies as well as mechanisms of resistance. Two multi-drug resistant and two sensitive serovars of Salmonella Weltevreden and Salmonella Newport isolated from poultry and seafood were used for growth kinetics and virulence gene expression study. The results obtained revealed that despite similar resistance pattern, effect of individual class of antibiotics on the growth of serovars varied. On the contrary, no significant difference was observed in growth pattern on exposure to these in vitro experimental conditions. Nevertheless, coupling these conditions with antibiotics drastically reduced the minimum inhibitory concentration (MIC) of antibiotics in resistant strains. A first of its kind study that draws attention on the significant effect of antibiotics and physiological conditions on MIC between resistant and sensitive non-typhoidal Salmonella serovars and expression of virulence genes from Salmonella pathogenicity island (SPI) 1 and 2 (invA, hilC, fliC2, sseA and ssrB).
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References
Brunelle BW, Bearson BL, Bearson S (2015) Chloramphenicol and tetracycline decrease motility and increase invasion and attachment gene expression in specific isolates of multidrug-resistant Salmonella enterica serovar typhimurium. Front Microbiol 5:801. https://doi.org/10.3389/fmicb.2014.00801
CDC (2020) Health information for international travel. Oxford University Press, New York. https://wwwnc.cdc.gov/travel/yellowbook/2020/travel-related-infectious-diseases/salmonellosis-nontyphoidal, https://www.cdc.gov/salmonella
Chen HM, Wang Y, Su LH et al (2013) Nontyphoid Salmonella infection: microbiology, clinical features, and antimicrobial therapy. Pediatr Neonatol 543:147–152. https://doi.org/10.1016/j.pedneo.2013.01.010
Chowdhury R, Sahu GK, Das J (1996) Stress response in pathogenic bacteria. J Biosci 21:149–160. https://doi.org/10.1007/BF02703105
Clements M, Eriksson S, Tezcan-Merdol D, Rhen M (2001) Virulence gene regulation in Salmonella enterica. Ann Med 33:178–185. https://doi.org/10.3109/07853890109002075
Clinical and Laboratory Standards Institute (2018) M07: Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically, 11th edn. Clinical and Laboratory Standards Institute, Wayne, PA, USA. ISBN 1562388363
Cummings LA, Wilkerson WD, Bergsbaken T et al (2006) In vivo, fliC expression by Salmonella enterica serovar typhimurium is heterogeneous, regulated by ClpX, and anatomically restricted. Mol Microbiol 61:795–809. https://doi.org/10.1111/j.1365-2958.2006.05271.x
Deekshit VK, Kumar BK, Rai P, Srikumar S et al (2012) Detection of class 1 integrons in Salmonella Weltevreden and silent antibiotic resistance genes in some seafood-associated nontyphoidal isolates of Salmonella in south-west coast of India. J Appl Microbiol 112:1113–1122. https://doi.org/10.1111/j.1365-2672.2012.05290.x
Deekshit VK, Kumar BK, Rai P, Karunasagar I, Karunasagar I (2015) Differential expression of virulence genes and role of gyrA mutations in quinolone resistant and susceptible strains of Salmonella Weltevreden and Newport isolated from seafood. J Appl Microbiol 119:970–980. https://doi.org/10.1111/jam.12924
Dichtl S, Demetz E, Haschka D et al (2019) Dopamine is a siderophore-like iron chelator that promotes Salmonella enterica serovar typhimurium virulence in mice. Mbio 10:e02624-e2718. https://doi.org/10.1128/mBio.02624-18
Fang FC, Frawley ER, Tapscott T et al (2014) Bacterial stress responses during host infection. Hysiol Behav 20:133–143. https://doi.org/10.1016/j.chom.2016.07.009
Foster JW, Spector MP (1995) How Salmonella survive against the odds. Annu Rev Microbiol 49:145–174. https://doi.org/10.1146/annurev.micro.49.1.145
Fridman O, Goldberg A, Ronin I, Shoresh N, Balaban NQ (2014) Optimization of lag time underlies antibiotic tolerance in evolved bacterial populations. Nature 513:418–421. https://doi.org/10.1038/nature13469
Gonzales-Siles L, Sjöling Å (2016) The different ecological niches of enterotoxigenic Escherichia coli. Environ Microbiol 18:741–751. https://doi.org/10.1111/1462-2920.13106
Gunasekera TS, Csonka LN, Paliy O (2008) Genome-wide transcriptional responses of Escherichia coli K-12 to continuous osmotic and heat stresses. J Bacteriol 190:3712–3720. https://doi.org/10.1128/JB.01990-07
Hong H, Jung J, Park W (2014) Plasmid-encoded tetracycline efflux pump protein alters bacterial stress responses and ecological fitness of Acinetobacter oleivorans. PLoS ONE 9(e107716):9. https://doi.org/10.1371/journal.pone.0107716
Jorgensen JH, Ferraro MJ (2009) Antimicrobial susceptibility testing: a review of general principles and contemporary practices. Clin Infect Dis 49:1749–1755
Kortman GA, Boleij A, Swinkels DW et al (2012) Iron availability increases the pathogenic potential of Salmonella typhimurium and other enteric pathogens at the intestinal epithelial interface. PLoS ONE 7:e29968. https://doi.org/10.1371/journal.pone.0029968
Kotian A, Aditya V, Jazeela K et al (2020) Effect of bile on growth and biofilm formation of non-typhoidal Salmonella serovars isolated from seafood and poultry. Res Microbiol 171:165–173. https://doi.org/10.1016/j.resmic.2020.06.002
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Majowicz SE, Musto J, Scallan E et al (2010) International collaboration on enteric disease “burden of illness” studies. The global burden of nontyphoidal Salmonella gastroenteritis. Clin Infect Dis 50:882–889. https://doi.org/10.1086/650733
Paiva JB, Cavallini JS, Silva MD et al (2009) Molecular differentiation of Salmonella Gallinarum and Salmonella Pullorum by RFLP of fliC gene from Brazilian isolates. Rev Bras 11:271–275. https://doi.org/10.1590/S1516-635X2009000400009
Rahn K, De Grandis SA, Clarke RC et al (1992) Amplification of an invA gene sequence of Salmonella typhimurium by polymerase chain reaction as a specific method of detection of Salmonella. Mol Cell Probes 6:271–279. https://doi.org/10.1016/0890-8508(92)90002-F
Schwarz S, Kehrenberg C, Doublet B, Cloeckaert A (2004) Molecular basis of bacterial resistance to chloramphenicol and florfenicol. FEMS Microbiol Rev 28:519–542. https://doi.org/10.1016/j.femsre.2004.04.001
Şimşek E, Kim M (2019) Power-law tail in lag time distribution underlies bacterial persistence. Proc Natl Acad Sci USA 116:17635–17640. https://doi.org/10.1073/pnas.1903836116
Spector MP, Kenyon WJ (2012) Resistance and survival strategies of Salmonella enterica to environmental stresses. Food Res Int 45:455–481. https://doi.org/10.1016/j.foodres.2011.06.056
Theophel K, Schacht VJ, Schlüter M et al (2014) The importance of growth kinetic analysis in determining bacterial susceptibility against antibiotics and silver nanoparticles. Front Microbiol 5:544. https://doi.org/10.3389/fmicb.2014.00544
Winfield MD, Groisman EA (2003) Role of nonhost environments in the lifestyles of Salmonella and Escherichia coli. Appl Environ Microbiol 69:3687–3694. https://doi.org/10.1128/AEM.69.7.3687
Yoon H, Park BY, Oh MH et al (2013) Effect of NaCl on heat resistance, antibiotic susceptibility, and Caco-2 cell invasion of Salmonella. Biomed Res Int 1:1–6. https://doi.org/10.1155/2013/274096
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The authors are thankful to Nitte University Centre for Science Education and Research for providing the necessary facilities and research fellowships.
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Financial support received from the Government of India funded DST-SERB (Grant no. ECR/2017/000559) toward this study is gratefully acknowledged.
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AK: Writing – original draft, performed the experiments, data analysis and wrote the paper. VA: Performed the experiments and formal analysis. JS: Performed the experiments and contributed to data curation. SS: Performed the experiments and contributed to data curation. PR: Data curation and contributed in data revision. AC: Writing- Reviewing and Editing and contributed in data revision. IK: Writing- Reviewing and Editing. VKD: Conceptualization, conceptualized the ideas and experimental design of the study and supervision.
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Kotian, A., Aditya, V., Sheikh, J. et al. Effect of NaCl, high iron, iron chelator and antibiotics on growth, virulence gene expression and drug susceptibility in non-typhoidal Salmonella: an in vitro fitness study. Arch Microbiol 204, 667 (2022). https://doi.org/10.1007/s00203-022-03278-x
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DOI: https://doi.org/10.1007/s00203-022-03278-x