New Strategies for Inhibition of Listeria monocytogenes and Klebsiella pneumoniae Biofilm Formation and Persistence


  • Samyah D. Jastaniah Department of Biology, College of Science, King Abdulaziz University, Jeddah, Saudi Arabia.
  • Taghreed Yasir Jamal Department of Biology, College of Science, King Abdulaziz University, Jeddah, Saudi Arabia.
  • Reda H. Amashah Department of Biology, College of Science, King Abdulaziz University, Jeddah, Saudi Arabia.
  • Magda M. Aly Department of Biology, College of Science, King Abdullaziz University, Jeddah, Saudi Arabia; Botany and Microbiology Department, Faculty of Science, Kafrelsheikh University, Egypt.



Biofilms, Plant Extracts, Exopolymeric, Listeria monocytogenes, Klebsiella, Pneumonia


Objective: This study aimed to find new strategies for the prevention of bacterial biofilms and investigate the effect of some plant extracts on the biofilm formation by certain pathogenic bacterial strains in-vitro.

Methods: Fourteen different biofilm forming bacterial isolates were collected   and their biofilm were quantitatively measured under different temperature, pH and growth medium using Crystal violet staining method.  Exopolysaccharides (EPS) produced by the isolates were estimated and a comparison between the tested isolates was made. The effect of some plant extracts on bacterial growth, biofilm formation and exopolysaccharide quantity was determined.

Results: The two isolates, Listeria monocytogenes (ATCC13932) and Klebsiella pneumoniae (ATCC700613) were among the most active biofilm forming bacterial isolates.  The optimum temperature, pH and media for EPS production and biofilm formation were determined. The effect of some plant extracts of Cranberry; Pomegranate peel and, Arak on growth and formation of biofilm and EPS were recorded. Moreover, minimum inhibition concentration of each plant extract was performed.

Conclusion: EPS quantity produced from the tested isolates depends on some effective factors such temperature, media contents and pH. Aqueous-Cranberry, aqueous-Pomegranate Peel and methanolic-Arak extracts have antibacterial and antibiofilm activities on L. monocytogenes and K.  pneumonia. Thus, application of new natural approaches for inhibiting bacterial biofilm is important to prevent persistent and recurrent biofilm related infections.


Abou-Zaid, A. A., Abd-Elmaguid, N. M., Abd El-Hafez, A., & Amer, M. M. (2015). The Effect of Arak Stems Extracts on Chemical Characteristics, Bacterial Activity and Sensory Evaluation of Beef Sausage Products. International Journal of Advances in Agricultural and Environmental Engg (IJAAEE), Vol 2.

Adesina, T., Nwinyi, O., & Olugbuyiro, J. (2015). Prevention of bacterial biofilms formation on urinary catheter by selected plant extracts. Pak. J. Biol. Sci, 18(2), 67-73.

Bazargani, M. M., & Rohloff, J. (2016). Antibiofilm activity of essential oils and plant extracts against Staphylococcus aureus and Escherichia coli biofilms. Food control, 61, 156-164.

Bhattacharjee, M. K., & Alenezi, T. (2020). Antibiotic in myrrh from Commiphora molmol preferentially kills nongrowing bacteria. Future science OA, 6(4), FSO458.

Borges, A., Abreu, A., Malheiro, J., Saavedra, M. J., & Simõe, M. (2013). Biofilm prevention and control by dietary phytochemicals. CECAV-Centro de Ciência Animal e Veterinária.

Christensen, G. D., Baldassarri, L., & Simpson, W. A. (1995). [38] Methods for studying microbial colonization of plastics. In Methods in Enzymology (Vol. 253, pp. 477-500). Academic Press.

Chusri, S., Sompetch, K., Mukdee, S., Jansrisewangwong, S., Srichai, T., Maneenoon, K., Voravuthikunchai, S. (2012). Inhibition of Staphylococcus epidermidis biofilm formation by traditional Thai herbal recipes used for wound treatment. Evidence-Based Complementary and Alternative Medicine, 2012.

de Oliveira, M. M. M., Brugnera, D. F., do Nascimento, J. A., Batista, N. N., & Piccoli, R. H. (2012). Cinnamon essential oil and cinnamaldehyde in the control of bacterial biofilms formed on stainless steel surfaces. European Food Research and Technology, 234(5), 821-832.

Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. t., & Smith, F. (1956). Colorimetric method for determination of sugars and related substances. Analytical chemistry, 28(3), 350-356.

Famuyide, I. M., Aro, A. O., Fasina, F. O., Eloff, J. N., & McGaw, L. J. (2019). Antibacterial and antibiofilm activity of acetone leaf extracts of nine under-investigated south African Eugenia and Syzygium (Myrtaceae) species and their selectivity indices. BMC Complementary and Alternative Medicine, 19(1), 141.

Guzman, J. P. M. D., De las Alas, T. P. L., Lucban, M. C., & Sevilla, C. E. C. (2020). Green tea (Camellia sinensis) extract inhibits biofilm formation in acyl homoserine lactone-producing, antibiotic-resistant Morganella morganii isolated from Pasig River, Philippines. Heliyon, 6(10), e05284.

Harjai, K., Bala, A., Gupta, R. K., & Sharma, R. (2013). Leaf extract of Azadirachta indica (neem): a potential antibiofilm agent for Pseudomonas aeruginosa. Pathogens and Disease, 69(1), 62-65.

Heatley, N. G. (1944). A method for the assay of penicillin. Biochemical Journal, 38(1), 61.

Hickl, J., Argyropoulou, A., Sakavitsi, M. E., Halabalaki, M., Al-Ahmad, A., Hellwig, E., Vach, K. (2018). Mediterranean herb extracts inhibit microbial growth of representative oral microorganisms and biofilm formation of Streptococcus mutans. PloS one, 13(12), e0207574.

Husain, F. M., Ahmad, I., Al-Thubiani, A. S., Abulreesh, H. H., AlHazza, I. M., & Aqil, F. (2017). Leaf extracts of Mangifera indica L. Inhibit quorum sensing–regulated production of virulence factors and biofilm in test bacteria. Frontiers in microbiology, 8, 727.

John, N. R., Gala, V. C., & Sawant, C. S. (2013). Inhibitory effects of plant extracts on multi-species dental biofilm formation in-vitro. Int J Pharm Bio Sci, 4(2), 487-495.

Kazemian, H., Ghafourian, S., Heidari, H., Amiri, P., Yamchi, J. K., Shavalipour, A., Sadeghifard, N. (2015). Antibacterial, anti-swarming and anti-biofilm formation activities of Chamaemelum nobile against Pseudomonas aeruginosa. Revista da Sociedade Brasileira de Medicina Tropical, 48(4), 432-436.

Lou, Z., Hong, Y., Liu, Y., Song, X., Ai, L., Wang, H., . . . Tang, Y. (2014). Effect of ethanol fraction of burdock leaf on biofilm formation and bacteria growth. European Food Research and Technology, 239(2), 305-311.

Mashhady, M. A., Abkhoo, J., Jahani, S., Abyar, S., & Khosravani, F. (2016). Inhibitory effects of plant extracts on Pseudomonas aeruginosa biofilm formation. International Journal of Infection, 3(4).

McLean, R. J., Pierson III, L. S., & Fuqua, C. (2004). A simple screening protocol for the identification of quorum signal antagonists. Journal of Microbiological Methods, 58(3), 351-360.

Millezi, F., Pereira, M. O., Batista, N., Camargos, N., Auad, I., Cardoso, M., & Piccoli, R. (2012). Susceptibility of monospecies and dual‐species biofilms of Staphylococcus aureus and Escherichia coli to essential oils. Journal of Food Safety, 32(3), 351-359.

Namasivayam, S. K. R., & Roy, E. A. (2013). Anti-biofilm effect of medicinal plant extracts against clinical isolate of biofilm of Escherichia coli. Int. J. Pharm. Pharm. Sci, 5(2), 486-489.

Nikolić, M., Vasić, S., Đurđević, J., Stefanović, O., & Čomić, L. (2014). Antibacterial and anti-biofilm activity of ginger (Zingiber officinale (Roscoe)) ethanolic extract. Kragujevac Journal of Science (36), 129-136.

Oh, S. K., Chang, H. J., Chun, H. S., Kim, H. J., & Lee, N. (2015). Pomegranate (Punica granatum L.) Peel Extract Inhibits Quorum Sensing and Biofilm Formation Potential in Yersinia enterocolitica. Microbiology and Biotechnology Letters, 43(4), 357-366.

Plyuta, V.A., Andreenko, J.V., Kuznetsov, A.E. et al. (2013). Formation of Pseudomonas aeruginosa PAO1 biofilms in the presence of hydrogen peroxide. The effect of the aiiA gene. Mol. Genet. Microbiol. Virol, 28, 141–146.

Quelemes, P. V., Perfeito, M. L., Guimarães, M. A., dos Santos, R. C., Lima, D. F., Nascimento, C., Eaton, P. (2015). Effect of neem (Azadirachta indica A. Juss) leaf extract on resistant Staphylococcus aureus biofilm formation and Schistosoma mansoni worms. Journal of ethnopharmacology, 175, 287-294.

Quave, C. L., Plano, L. R., Pantuso, T., & Bennett, B. C. (2008). Effects of extracts from Italian medicinal plants on planktonic growth, biofilm formation and adherence of methicillin-resistant Staphylococcus aureus. Journal of ethnopharmacology, 118(3), 418-428.

Sadekuzzaman, M., Yang, S., Mizan, M., & Ha, S. (2015). Current and recent advanced strategies for combating biofilms. Comprehensive Reviews in Food Science and Food Safety, 14(4), 491-509.

Sandasi, M., Leonard, C., Van Vuuren, S., & Viljoen, A. (2011). Peppermint (Mentha piperita) inhibits microbial biofilms in vitro. South African Journal of Botany,77(1), 80-85.

Satpathy, S., Sen, S. K., Pattanaik, S., & Raut, S. (2016). Review on bacterial biofilm: A universal cause of contamination. Biocatalysis and agricultural biotechnology, 7, 56-66.

Schneider-Rayman, M., Steinberg, D., Sionov, R. V., Friedman, M., & Shalish, M. (2021). Effect of epigallocatechin gallate on dental biofilm of Streptococcus mutans: An in vitro study. BMC oral health, 21(1), 1-11.

Semeniuc, C. A., Pop, C. R., & Rotar, A. M. (2017). Antibacterial activity and interactions of plant essential oil combinations against Gram-positive and Gram-negative bacteria. journal of food and drug analysis, 25(2), 403-408.

Smitinont, T., Tansakul, C., Tanasupawat, S., Keeratipibul, S., Navarini, L., Bosco, M., & Cescutti, P. (1999). Exopolysaccharide-producing lactic acid bacteria strains from traditional thai fermented foods: isolation, identification and exopolysaccharide characterization. International journal of food microbiology, 51(2-3), 105-111.

Ulrey, R. K., Barksdale, S. M., Zhou, W., & van Hoek, M. L. (2014). Cranberry proanthocyanidins have anti-biofilm properties against Pseudomonas aeruginosa. BMC Complementary and Alternative Medicine, 14(1), 1-12.

Wayne, PA. (2017). Clinical and Laboratory Standards Institute. Performance Standards for 205 Antimicrobial Susceptibility Testing: 27th Informational Supplement. M100-S27. Clinical 206 and Laboratory Standards Institute.

Wojnicz, D., Kucharska, A. Z., Sokół-Łętowska, A., Kicia, M., & Tichaczek-Goska, D. (2012). Medicinal plants extracts affect virulence factors expression and biofilm formation by the uropathogenic Escherichia coli. Urological Research, 40(6), 683-697.




How to Cite

Jastaniah, S. D. ., Jamal, T. Y. ., Amashah, R. H. ., & Aly, M. M. . (2022). New Strategies for Inhibition of Listeria monocytogenes and Klebsiella pneumoniae Biofilm Formation and Persistence. Journal of Contemporary Medical Sciences, 8(6), 400–407.