The effect of Lactiplantibacillus plantarum and Lacticaseibacillus rhamnosus cell-free supernatant on mature polymicrobial biofilm mass of Candida albicans and Escherichia coli

Kadek Indah Novitasari Sudarsana; Rebekah Juniati Setiabudi; Agung Dwi Wahyu Widodo

doi:10.31893/multiscience.2025241

Keywords:

Polymicrobial biofilm

L. rhamnosus

L. plantarum

C. albicans

E. coli

Abstract

Biofilm is one of the leading causes of prolonged infection known currently with higher treatment failure if caused by polymicrobial biofilm. Lactobacillus strain is known to produce antibiofilm substances that could disrupt biofilm by fungi and bacteria. This study aims to evaluate two probiotic strains, L. plantarum and L. rhamnosus’ ability as single and multistrain to disrupt the biofilm of Candida albicans and Escherichia coli. This study employed a true experiment post test only control group design, using C. albicans and E. coli clinical isolate as subjects. The subjects in this study are divided into control and treatment groups. Treatment groups were subdivided into those receiving L. plantarum, L. rhamnosus and both probiotics for 24 h. Pathogens were incubated for 48 h in a 96-well microplate to reach a mature state before treatment groups were exposed to probiotics suspension for 24 h. Then, biomass was monitored using Crystal Violet (CV) staining and read using an ELISA reader to obtain the Optical Density (OD) values. The biofilm mass between treatments and control groups demonstrated a meaningful difference (p < 0.05), further post hoc analysis demonstrated multistrain cell-free supernatant of L. plantarum and L. rhamnosus demonstrated the highest antibiofilm between treatment groups in lowering biomass of single and dual-species biofilm C. albicans and E. coli (C. albicans = 0.215 ± 0.086; E. coli = 0.218 ± 0.099; dual-species biofilm = 0.123 ± 0.025). With the exposure of 24 h, the combination of L. plantarum and L. rhamnosus can decrease the biomass of single or dual-species biofilm formed by C. albicans and E. coli.
References
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19. Matsuda, Y., Cho, O., Sugita, T., Ogishima, D., & Takeda, S. (2018). Culture Supernatants of Lactobacillus gasseri and L. crispatus Inhibit Candida albicans Biofilm Formation and Adhesion to HeLa Cells. Mycopathologia, 183 (4): 691-700. https://doi.org/10.1007/s11046-018-0259-4
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21. Murtiastutik, D., Emma Hidayati Sasmito, E., Afif Nurul Hidayati, A., Utomo, B., Endraswari, P. D., & Indramaya, D. M. (2022). Basil leaf extract and eugenol against isolates of Candida sp. causing oral candidiasis in HIV/AIDS. Journal of Pakistan Association of Dermatologists, 32 (4): 669-675.
22. Nobile, C. J., Schneider, H. A., Nett, J. E., Sheppard, D. C., Filler, S. G., Andes, D. R., & Mitchell, A. P. (2008). Complementary adhesin function in C. albicans biofilm formation. Current Biology, 18 (14): 1017-1024. https://doi.org/10.1016/j.cub.2008.06.034
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29. Vazquez-Munoz, R., & Dongari-Bagtzoglou, A. (2021). Anticandidal Activities by Lactobacillus Species: An Update on Mechanisms of Action. Front Oral Health, 2: 689382. https://doi.org/10.3389/froh.2021.689382
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32. Zeise, K. D., Woods, R. J., & Huffnagle, G. B. (2021). Interplay between Candida albicans and Lactic Acid Bacteria in the Gastrointestinal Tract: Impact on Colonization Resistance, Microbial Carriage, Opportunistic Infection, and Host Immunity. Clinical Microbiology Reviews, 34 (4): e0032320. https://doi.org/10.1128/cmr.00323-20

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How to cite

Sudarsana, K. I. N., Setiabudi, R. J., & Widodo, A. D. W. (2024). The effect of Lactiplantibacillus plantarum and Lacticaseibacillus rhamnosus cell-free supernatant on mature polymicrobial biofilm mass of Candida albicans and Escherichia coli. Multidisciplinary Science Journal, 7(5), 2025241. https://doi.org/10.31893/multiscience.2025241

[1] Ahmed, S. S., Shariq, A., Alsalloom, A. A., Babikir, I. H., & Alhomoud, B. N. (2019). Uropathogens and their antimicrobial resistance patterns: Relationship with urinary tract infections. Int J Health Sci (Qassim), 13 (2): 48-55.

[2] Azevedo, A. S., Almeida, C., Melo, L. F., & Azevedo, N. F. (2017). Impact of polymicrobial biofilms in catheter-associated urinary tract infections. Critical Reviews in Microbiology, 43 (4): 423-439. https://doi.org/10.1080/1040841x.2016.1240656

[3] Bjarnsholt, T. (2011). Introduction to biofilms. Biofilm infections: 1-9.

[4] Carvalho, F. M., Mergulhão, F. J. M., & Gomes, L. C. (2021). Using Lactobacilli to Fight Escherichia coli and Staphylococcus aureus Biofilms on Urinary Tract Devices. Antibiotics (Basel), 10 (12). https://doi.org/10.3390/antibiotics10121525

[5] De Brucker, K., Tan, Y., Vints, K., De Cremer, K., Braem, A., Verstraeten, N., Michiels, J., Vleugels, J., Cammue, B. P., & Thevissen, K. (2015). Fungal β-1,3-glucan increases ofloxacin tolerance of Escherichia coli in a polymicrobial E. coli/Candida albicans biofilm. Antimicrobial Agents and Chemotherapy, 59 (6): 3052-3058. https://doi.org/10.1128/aac.04650-14

[6] Delcaru, C., Alexandru, I., Podgoreanu, P., Grosu, M., Stavropoulos, E., Chifiriuc, M. C., & Lazar, V. (2016). Microbial Biofilms in Urinary Tract Infections and Prostatitis: Etiology, Pathogenicity, and Combating strategies. Pathogens, 5 (4). https://doi.org/10.3390/pathogens5040065

[7] Ervianti, E., Damayanti, Purnamasari, I., Astari, L., Prasetyo, B., Endraswari, P. D., Utomo, B., Fitriani, E. W., Indramaya, D. M., Listiawan, M. Y., & Sigit Prakoeswa, C. R. (2023). Comparison of tea tree oil 5%, tea tree oil 10%, and nystatin inhibition zones against vaginal Candida isolates in pregnancy. J Infect Dev Ctries, 17 (3): 353-358. https://doi.org/10.3855/jidc.16761

[8] Eshima, S., Kurakado, S., Matsumoto, Y., Kudo, T., & Sugita, T. (2022). Candida albicans Promotes the Antimicrobial Tolerance of Escherichia coli in a Cross-Kingdom Dual-Species Biofilm. Microorganisms, 10 (11). https://doi.org/10.3390/microorganisms10112179

[9] Farrokhi, Y., Al-Shibli, B., Al-Hameedawi, D. F., Neshati, Z., & Makhdoumi, A. (2021). Escherichia coli enhances the virulence factors of Candida albicans, the cause of vulvovaginal candidiasis, in a dual bacterial/fungal biofilm. Research in Microbiology, 172 (4-5): 103849. https://doi.org/10.1016/j.resmic.2021.103849

[10] Gaston, J. R., Johnson, A. O., Bair, K. L., White, A. N., & Armbruster, C. E. (2021). Polymicrobial interactions in the urinary tract: is the enemy of my enemy my friend? Infect Immun. https://doi.org/10.1128/iai.00652-20

[11] Gulati, M., Lohse, M. B., Ennis, C. L., Gonzalez, R. E., Perry, A. M., Bapat, P., Arevalo, A. V., Rodriguez, D. L., & Nobile, C. J. (2018). In Vitro Culturing and Screening of Candida albicans Biofilms. Current Protocols in Microbiology, 50 (1): e60. https://doi.org/10.1002/cpmc.60

[12] Gulati, M., & Nobile, C. J. (2016). Candida albicans biofilms: development, regulation, and molecular mechanisms. Microbes Infect, 18 (5): 310-321. https://doi.org/10.1016/j.micinf.2016.01.002

[13] Hill, C., Guarner, F., Reid, G., Gibson, G. R., Merenstein, D. J., Pot, B., Morelli, L., Canani, R. B., Flint, H. J., Salminen, S., Calder, P. C., & Sanders, M. E. (2014). Expert consensus document. The International Scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nature Reviews: Gastroenterology & Hepatology, 11 (8): 506-514. https://doi.org/10.1038/nrgastro.2014.66

[14] Itapary Dos Santos, C., Ramos França, Y., Duarte Lima Campos, C., Quaresma Bomfim, M. R., Oliveira Melo, B., Assunção Holanda, R., Santos, V. L., Gomes Monteiro, S., Buozzi Moffa, E., Souza Monteiro, A., Andrade Monteiro, C., & Monteiro-Neto, V. (2019). Antifungal and Antivirulence Activity of Vaginal Lactobacillus Spp. Products against Candida Vaginal Isolates. Pathogens, 8 (3). https://doi.org/10.3390/pathogens8030150

[15] Lila, A. S. A., Rajab, A. A. H., Abdallah, M. H., Rizvi, S. M. D., Moin, A., Khafagy, E. S., Tabrez, S., & Hegazy, W. A. H. (2023). Biofilm Lifestyle in Recurrent Urinary Tract Infections. Life (Basel), 13 (1). https://doi.org/10.3390/life13010148

[16] Liu, Y., & Filler, S. G. (2011). Candida albicans Als3, a multifunctional adhesin and invasin. Eukaryotic Cell, 10 (2): 168-173. https://doi.org/10.1128/ec.00279-10

[17] Maione, A., Galdiero, E., Cirillo, L., Gambino, E., Gallo, M. A., Sasso, F. P., Petrillo, A., Guida, M., & Galdiero, M. (2023). Prevalence, Resistance Patterns and Biofilm Production Ability of Bacterial Uropathogens from Cases of Community-Acquired Urinary Tract Infections in South Italy. Pathogens, 12 (4). https://doi.org/10.3390/pathogens12040537

[18] Mani-López, E., Arrioja-Bretón, D., & López-Malo, A. (2022). The impacts of antimicrobial and antifungal activity of cell-free supernatants from lactic acid bacteria in vitro and foods. Compr Rev Food Sci Food Saf, 21 (1): 604-641. https://doi.org/10.1111/1541-4337.12872

[19] Matsuda, Y., Cho, O., Sugita, T., Ogishima, D., & Takeda, S. (2018). Culture Supernatants of Lactobacillus gasseri and L. crispatus Inhibit Candida albicans Biofilm Formation and Adhesion to HeLa Cells. Mycopathologia, 183 (4): 691-700. https://doi.org/10.1007/s11046-018-0259-4

[20] Medina, M., & Castillo-Pino, E. (2019). An introduction to the epidemiology and burden of urinary tract infections. Therapeutic Advances in Urology, 11: 1756287219832172. https://doi.org/10.1177/1756287219832172

[21] Murtiastutik, D., Emma Hidayati Sasmito, E., Afif Nurul Hidayati, A., Utomo, B., Endraswari, P. D., & Indramaya, D. M. (2022). Basil leaf extract and eugenol against isolates of Candida sp. causing oral candidiasis in HIV/AIDS. Journal of Pakistan Association of Dermatologists, 32 (4): 669-675.

[22] Nobile, C. J., Schneider, H. A., Nett, J. E., Sheppard, D. C., Filler, S. G., Andes, D. R., & Mitchell, A. P. (2008). Complementary adhesin function in C. albicans biofilm formation. Current Biology, 18 (14): 1017-1024. https://doi.org/10.1016/j.cub.2008.06.034

[23] Pertiwi, M. I. C., Wahyunitisari, M. R., Setiabudi, R. J., Widodo, A. D. W., Wiqoyah, N., & Arfijanto, M. V. (2023). Inhibitory effects of synbiotics on biofilm of uropathogenic Escherichia coli during urinary tract infection. Journal of Advanced Biotechnology and Experimental Therapeutics, 6 (2): 350-4760.

[24] Pinto, H., Simões, M., & Borges, A. (2021). Prevalence and Impact of Biofilms on Bloodstream and Urinary Tract Infections: A Systematic Review and Meta-Analysis. Antibiotics (Basel), 10 (7). https://doi.org/10.3390/antibiotics10070825

[25] Rishpana, M. S., & Kabbin, J. S. (2015). Candiduria in Catheter Associated Urinary Tract Infection with Special Reference to Biofilm Production. J Clin Diagn Res, 9 (10): Dc11-13. https://doi.org/10.7860/jcdr/2015/13910.6690

[26] Song, H., Zhang, J., Qu, J., Liu, J., Yin, P., Zhang, G., & Shang, D. (2019). Lactobacillus rhamnosus GG microcapsules inhibit Escherichia coli biofilm formation in coculture. Biotechnol Lett, 41 (8-9): 1007-1014. https://doi.org/10.1007/s10529-019-02694-2

[27] Stepanović, S., Vuković, D., Hola, V., Di Bonaventura, G., Djukić, S., Cirković, I., & Ruzicka, F. (2007). Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. APMIS, 115 (8): 891-899. https://doi.org/10.1111/j.1600-0463.2007.apm_630.x

[28] Vagios, S., Hesham, H., & Mitchell, C. (2020). Understanding the potential of lactobacilli in recurrent UTI prevention. Microbial Pathogenesis, 148: 104544. https://doi.org/10.1016/j.micpath.2020.104544

[29] Vazquez-Munoz, R., & Dongari-Bagtzoglou, A. (2021). Anticandidal Activities by Lactobacillus Species: An Update on Mechanisms of Action. Front Oral Health, 2: 689382. https://doi.org/10.3389/froh.2021.689382

[30] Wang, C., Chang, T., Yang, H., & Cui, M. (2015). Antibacterial mechanism of lactic acid on physiological and morphological properties of Salmonella Enteritidis, Escherichia coli and Listeria monocytogenes. Food Control, 47: 231–236. https://doi.org/10.1016/j.foodcont.2014.06.034

[31] Yang, W., Zhou, Y., Wu, C., & Tang, J. (2016). Enterohemorrhagic Escherichia coli promotes the invasion and tissue damage of enterocytes infected with Candida albicans in vitro. Scientific Reports, 6 (1): 37485. https://doi.org/10.1038/srep37485

[32] Zeise, K. D., Woods, R. J., & Huffnagle, G. B. (2021). Interplay between Candida albicans and Lactic Acid Bacteria in the Gastrointestinal Tract: Impact on Colonization Resistance, Microbial Carriage, Opportunistic Infection, and Host Immunity. Clinical Microbiology Reviews, 34 (4): e0032320. https://doi.org/10.1128/cmr.00323-20

Abstract

References

How to cite