Cinnamaldehyde: a compound with antimicrobial and synergistic activity against ESBL-producing quinolone-resistant pathogenic Enterobacteriaceae

  • Lena Dhara
  • Anusri TripathiEmail author
Original Article


Usage of cephalosporin and quinolone antibiotics has aggravated the development of extended-spectrum beta-lactamase (ESBL)–producing quinolone-resistant (QR) pathogenic Enterobacteriaceae. The present study aims to determine antimicrobial activity of cinnamaldehyde alone or in combination with cefotaxime/ciprofloxacin to reverse the drug resistance and evaluations of efficacy, and possible molecular mechanism of action of the combination was also evaluated using in vitro assays. Broth microdilution assay was used to determine minimum inhibitory concentrations (MICs) of cinnamaldehyde and antibiotics against ESBL-QR Enterobacteriaceae. Synergistic effect and dynamic interaction with antibiotics were further examined by checkerboard assay, isobologram analysis, and time-kill assay, respectively. Cellular morphology of bacteria was viewed with scanning electron microscopy (SEM). Effects of cinnamaldehyde and its combination on the expression of gene encoding—porins (ompC, ompF, ompK35, and ompK36), efflux pump genes (acrB–E. coli, acrB–K. pneumoniae), and antibiotic-resistant genes (blaTEM, blaSHV, blaCTXM, and QnrB) were evaluated using real-time quantitative PCR (RT-qPCR). Majority of the E. coli (32.1%) and K. pneumoniae (24.2%) isolates demonstrated MIC of cinnamaldehyde at 7.34 μg/mL and 0.91 g/mL, respectively. Synergism between cinnamaldehyde and cefotaxime was noted among 75% E. coli and 60.6% K. pneumoniae. Similarly, synergism with ciprofloxacin was observed among 39.6% and 42.4% of the bacteria, respectively. Thus, cinnamaldehyde reduced MIC of cefotaxime and ciprofloxacin 2–1024-fold with bactericidal and/synergistic effect after 24 h. Cinnamaldehyde and its combination altered gene expression by ~ 1.6 to ~ 400-fold. Distorted bacterial cell structures were visible after treatment with cinnamaldehyde and/or with cefotaxime/ciprofloxacin. The results indicated the potential efficacy and mode of action of cinnamaldehyde alone and in combination with antibiotics against pathogenic ESBL-QR bacteria.


Cinnamaldehyde ESBL Quinolone Synergy Scanning electron microscopy Real-time quantitative PCR 



The authors are extremely grateful to the Director, Calcutta School of Tropical Medicine, Kolkata, India, for providing necessary facilities for this study.

Funding information

This study is funded by the Indian Council of Medical Research (Grant No. -58/67/BMS-2012).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical statement and informed consent

The study was approved by the ethical research committee (reference number: CREC-STM/53 dated 23/09/2011). Informed consent was obtained from the patients for participating in the study.

Supplementary material

10096_2019_3692_MOESM1_ESM.docx (16 kb)
ESM 1 (DOCX 15 kb)
10096_2019_3692_MOESM2_ESM.doc (116 kb)
ESM 2 (DOC 116 kb)
10096_2019_3692_MOESM3_ESM.doc (34 kb)
ESM 3 (DOC 33 kb)


  1. 1.
    Chandra H, Bishnoi P, Yadav A, Patni B, Mishra AP, Nautiyal AR (2017) Antimicrobial resistance and he alternative resources with special emphasis on plant-based antimicrobials-a review. Plants (Basel) 10(2):6Google Scholar
  2. 2.
    Frank T, Mbecko JR, Misatou P, Monchy D (2011) Emergence of quinolone resistance among extended-spectrum beta-lactamase-producing Enterobacteriaceae in the Central African Republic: genetic characterization. BMC Res Notes 25:309CrossRefGoogle Scholar
  3. 3.
    Laxminarayan R, Chaudhury RR (2016) Antibiotic resistance in India: drivers and opportunities for action. PLoS Med 13:e1001974CrossRefGoogle Scholar
  4. 4.
    Alekshun MN, Stuart B (2007) Molecular mechanisms of antibacterial multidrug resistance. Cell 128:1037–1050CrossRefGoogle Scholar
  5. 5.
    van Hoek Angela HAM, Mevius D, Guerra B, Mullany P, Roberts Paul A, Aarts Henk JM (2011) Acquired antibiotic resistance genes: an overview. Front Microbiol 2:1–27Google Scholar
  6. 6.
    Atanasov AG, Waltenberger B, Pferschy-Wenzig EM, Linder T, Wawrosch C, Uhrin P et al (2015) Discovery and resupply of pharmacologically active plant-derived natural products: a review. Biotechnol Adv 33:1582–1614CrossRefGoogle Scholar
  7. 7.
    Jia P, Xue YJ, Duan XJ, Shao SH (2011) Effect of cinnamaldehyde on biofilm formation and sarA expression by methicillin-resistant Staphylococcus aureus. Lett Appl Microbiol 53:409–416CrossRefGoogle Scholar
  8. 8.
    López P, Sanchez C, Batlle R, Nerín C (2007) Vapor-phase activities of cinnamon, thyme, and oregano essential oils and key constituents against foodborne microorganisms. J Agric Food Chem 55:4348–4356CrossRefGoogle Scholar
  9. 9.
    Liu Q, Niu H, Zhang W, Mu H, Sun C, Duan J (2015) Synergy among thymol, eugenol, berberine, cinnamaldehyde and streptomycin against planktonic and biofilm-associated food-borne pathogens. Lett Appl Microbiol 60:421–430CrossRefGoogle Scholar
  10. 10.
    Palaniappan K, Holley RA (2010) Use of natural antimicrobials to increase antibiotic susceptibility of drug resistant bacteria. Int J Food Microbiol 140:64–68CrossRefGoogle Scholar
  11. 11.
    Hossan MS, Jindal H, Maisha S, Samudi Raju C, Devi Sekaran S, Nissapatorn V, Kaharudin F, Su Yi L, Khoo TJ, Rahmatullah M, Wiart C (2018) Antibacterial effects of 18 medicinal plants used by the Khyang tribe in Bangladesh. Pharm Biol 56:201–208CrossRefGoogle Scholar
  12. 12.
    Clinical and Laboratory Standards Institute (2008) Performance standards for antimicrobial susceptibility testing; eighteenth informational supplement, Document M100-S18. CLSI, Wayne, PAGoogle Scholar
  13. 13.
    Garcia L (2010) Synergism testing: broth microdilution checkerboard and broth macrodilution methods. In: Garcia L (ed) Clinical microbiology procedures handbook, 3rd edn. ASM Press, Washington, pp 140–162Google Scholar
  14. 14.
    Chou TC (2006) Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacolo Rev 58:31Google Scholar
  15. 15.
    Zhou XY, Ye XG, He LT, Zhang SR, Wang RL, Zhou J, He ZS (2016) In vitro characterization and inhibition of the interaction between ciprofloxacin and berberine against multidrug-resistant Klebsiella pneumoniae. J Antibiot (Tokyo) 69:741–746CrossRefGoogle Scholar
  16. 16.
    Salmi C, Loncle C, Vidal N, Letourneux Y, Fantini J, Maresca M, Taïeb N, Pagès JM, Brunel JM (2008) Squalamine: an appropriate strategy against the emergence of multidrug resistant gram-negative bacteria? PLoS One 23:e2765CrossRefGoogle Scholar
  17. 17.
    Cui Y, Kim SH, Kim H, Yeom J, Ko K et al (2012) AFM probing the mechanism of synergistic effects of the green tea polyphenol (2)-epigallocatechin-3-gallate (EGCG) with cefotaxime against extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli. PLoS One 7:e48880CrossRefGoogle Scholar
  18. 18.
    Justice SS, Hunstad DA, Cegelski L, Hultgren SJ (2008) Morphological plasticity as a bacterial survival strategy. Nature 6:162–168Google Scholar
  19. 19.
    Bos J, Zhang Q, Vyawahare S, Rogers E, Rosenberg SM, Austin RH (2015) Emergence of antibiotic resistance from multinucleated bacterial filaments. Proc Natl Acad Sci U S A 112:178–183CrossRefGoogle Scholar
  20. 20.
    Wang Y, Zhang Y, Shi YQ, Pan XH, Lu YH, Cao P (2018) Antibacterial effects of cinnamon (Cinnamomum zeylanicum) bark essential oil on Porphyromonas gingivalis. Microb Pathog 116:26–32CrossRefGoogle Scholar
  21. 21.
    Helander IM, Alakomi HL, Kyösti KL, Mattila-Sandholm T, Pol I, Smid EJ (1998) Characterization of the action of selected essential oil components on gram-negative bacteria. J Agric Food Chem 46:3590–3595CrossRefGoogle Scholar
  22. 22.
    Shen S, Zhang T, Yuan Y, Lin S, Xu J, Ye H (2015) Effects of cinnamaldehyde on Escherichia coli and Staphylococcus aureus membrane. Food Control 47:196–202CrossRefGoogle Scholar
  23. 23.
    García-Salinas S, Elizondo-Castillo H, Arruebo M, Mendoza G, Irusta S (2018) Evaluation of the antimicrobial activity and cytotoxicity of different components of natural origin present in essential oils. Molecules 23:1399CrossRefGoogle Scholar
  24. 24.
    Raja SB, Murali MR, Devaraj SN (2008) Differential expression of OmpC and OmpF in multidrug-resistant Shigella dysenteriae and Shigella flexneri by aqueous extract of Aeglemarmelos, altering its susceptibility toward beta-lactam antibiotics. Diagn Microbiol Infect Dis 61:321–328CrossRefGoogle Scholar
  25. 25.
    Raja SB, Murali MR, Malathi GK, Anbarasu K, Devaraj SN (2009) Effect of aqueous extract of Aegle marmelos fruit on adherence and β-lactam resistance of Enteropathogenic Escherichia coli by down regulating outer membrane protein C. Am J Infect Dis 5:161–169CrossRefGoogle Scholar
  26. 26.
    Karumathil DP, Nair MS, Gaffney J, Kollanoor-Johny A, Venkitanarayanan K (2018) Trans-cinnamaldehyde and eugenol increase Acinetobacter baumannii sensitivity to beta-lactam antibiotics. Front Microbiol 9:1011CrossRefGoogle Scholar
  27. 27.
    Zhou X, Jia F, Liu X, Wang Y (2012) Total alkaloids of Sophorea alopecuroides-induced down-regulation of AcrAB-TolC efflux pump reverses susceptibility to ciprofloxacin in clinical multidrug resistant Escherichia coli isolates. Phytother Res 261637–43Google Scholar
  28. 28.
    Cai W, Fu Y, Zhang W, Chen X, Zhao J, Song W et al (2016) Synergistic effects of baicalein with cefotaxime against Klebsiella pneumoniae through inhibiting CTX-M-1 gene expression. BMC Microbiol 16:181CrossRefGoogle Scholar
  29. 29.
    National Toxicology Program (2004) NTP toxicology and carcinogenesis studies of trans-cinnamaldehyde (CAS no. 14371-10-9) in F344/N rats and B6C3F1 mice (feed studies). Natl Toxicol Program Tech Rep Ser 514:1–281Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Biochemistry and Medical BiotechnologyCalcutta School of Tropical MedicineKolkataIndia

Personalised recommendations