The effect of probiotic Lactobacillus rhamnosus GG on Candida albicans and Candida tropicalis biofilm  formation: A preliminary study

Aalina Mohd Fuad; Wan Nur Akma Wan Mazlan; Ridhwan Abdul Wahab; Nurrul Shaqinah Nasruddin; Mohd Hafiz Arzmi

doi:10.31436/ijohs.v4i2.204

Authors

Aalina Mohd Fuad Cluster of Cancer Research Initiative IIUM (COCRII), International Islamic University Malaysia, Kuantan Campus, Pahang, Malaysia & Jardin Pharma Berhad, Sunway Subang Business Park, 40150 Shah Alam, Selangor, Malaysia.
Wan Nur Akma Wan Mazlan Cluster of Cancer Research Initiative IIUM (COCRII), International Islamic University Malaysia, Kuantan Campus, Pahang, Malaysia & Jardin Pharma Berhad, Sunway Subang Business Park, 40150 Shah Alam, Selangor, Malaysia.
Ridhwan Abdul Wahab International Medical School, Management and Science University (MSU), Section 13, 40100 Shah Alam, Selangor, Malaysia
Nurrul Shaqinah Nasruddin Department of Craniofacial Diagnostics and Biosciences, Faculty of Dentistry, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.
Mohd Hafiz Arzmi Cluster of Cancer Research Initiative IIUM (COCRII), International Islamic University Malaysi & Jardin Pharma Berhad, Sunway Subang Business Park, 40150 Shah Alam, Selangor, Malaysia & Department of Fundamental Dental and Medical Sciences, Kulliyyah of Dentistry, International Islamic University Malaysia, Kuantan Campus, Pahang, Malaysia.

DOI:

https://doi.org/10.31436/ijohs.v4i2.204

Keywords:

biofilm, Candida albicans, Candida tropicalis, Lactobacillus rhamnosus GG, probiotic

Abstract

Lactobacillus rhamnosus are Gram-positive and lactic acid-producing bacteria. Meanwhile, Candida albicans and Candida tropicalis are opportunistic fungi that cause oral candidiasis. This study aimed to determine the effect of LGG on the biofilm formation of C. tropicalis and C. albicans with the hypothesis that LGG inhibits the biofilm of the yeasts. C. albicans ATCC MYA-4901 and C. tropicalis ATCC 13803 were standardised to 1x106 cells to form a mono-species biofilm. L. rhamnosus GG (LGG) was standardised to 1x10⁷ cells, equivalent to absorbance 0.5 at OD620nm. The microorganisms were cultivated in nutrient broth in a 96-well plate and incubated at 37°C for 24 h and 48 h. Co-culture biofilm was developed by combining Candida spp. with LGG in the same well at a similar concentration as the mono-culture. Crystal violet assay was conducted to assess the biofilm biomass with absorbance measured at OD_620nm wavelength. After 24 hours, polymicrobial biofilms of C. albicans with LGG decreased by 37.1 ± 9.2%. At 48 hours, it further decreased to 44.7 ± 5.9%. For C. tropicalis, co-culture biofilms with LGG decreased by 16.3 ± 5.9% and 35.7 ± 7.6% after 24 h and 48 h incubation, respectively. LGG significantly reduced C. albicans biofilm compared to C. tropicalis (P<0.05). In conclusion, LGG has antibiofilm activity against C. albicans and C. tropicalis. However, further study is needed to conclude the effect against other species strains.

References

Alnuaimi, A. D., et al. (2013) Clinical isolates and laboratory reference Candida species and strains have varying abilities to form biofilms. FEMS Yeast Research, 13, 689– 699.

Alok, A., et al. (2017). Probiotics: A new era of biotherapy. Advanced Biomedical Research, 6, 31-31. https://doi.org/10.4103/2277-9175.192625

Andes, D., et al. (2004). Development and characterization of an in vivo central venous catheter Candida albicans biofilm model. Infection and immunity, 72(10), 6023-6031. https://doi.org/10.1128/iai.72.10.6023-6031.2004

Badri, P. E. A. M., et al. (2022). Characterization of Cervus timorensis velvet antler and its effect on biofilm formation of Candida species. Medical Mycology, 60(9), myac073.

Barzegari, A., et al. (2020). The battle of probiotics and their derivatives against biofilms. Infection and drug resistance, 13, 659.

Cavalheiro, M., et al. (2018). Candida Biofilms: threats, challenges, and promising strategies. Frontiers in Medicine, 5(28). https://doi.org/10.3389/fmed.2018.00028

Chen, L., et al. (2019). Lactobacillus rhamnosus GG treatment improves intestinal permeability and modulates microbiota dysbiosis in an experimental model of sepsis. International Journal of Molecular Medicine. https://doi.org/10.3892/ijmm.2019.4050

Corr, S. C., et al. (2009). Understanding the mechanisms by which probiotics inhibit gastrointestinal pathogens. Advances in Food and Nutrition Research, 56, 1-15.

Dimitrijevic, R., et al. (2009). The identification of a low molecular mass bacteriocin, rhamnosin A, produced by Lactobacillus rhamnosus strain 68. Journal of Applied Microbiology, 107(6), 2108-2115. https://doi.org/https://doi.org/10.1111/j.1365-2672.2009.04539.x

Doron, S., et al. (2005). Lactobacillus GG: Bacteriology and Clinical Applications. Gastroenterology Clinics, 34(3), 483-498. https://doi.org/10.1016/j.gtc.2005.05.011

Erdogan, A., et al. (2015). Small Intestinal Fungal Overgrowth. Current Gastroenterology Reports, 17(4), 16. https://doi.org/10.1007/s11894-015-0436-2

Gao, J., et al. (2019). A Novel Postbiotic From Lactobacillus rhamnosus GG with a beneficial effect on intestinal barrier function. Frontiers in Microbiology, 10(477). https://doi.org/10.3389/fmicb.2019.00477

Geng, M., et al. (2018). Covalent structure and bioactivity of the type aii Lantibiotic Salivaricin A2. Applied and Environmental Microbiology, 84(5), e02528-02517. https://doi.org/10.1128/AEM.02528-17

Hernday, A. D., et al. (2010). Genetics and molecular biology in Candida albicans. In Methods in Enzymology (Vol. 470, pp. 737-758). Academic Press. https://doi.org/https://doi.org/10.1016/S0076-6879(10)70031-8

Hu, L., et al. (2019). Characterization of oral candidiasis and the Candida species profile in patients with oral mucosal diseases. Microbial Pathogenesis, 134, 103575. https://doi.org/10.1016/j.micpath.2019.103575

Jiang, Q., et al. (2016). Interactions between Lactobacillus rhamnosus GG and oral micro-organisms in an in vitro biofilm model. BMC Microbiology, 16(1), 149. https://doi.org/10.1186/s12866-016-0759-7

Jørgensen, M. R., et al. (2017). Probiotic Lactobacillus reuteri has antifungal effects on oral Candida species in vitro. Journal of Oral Microbiology, 9(1), 1274582. https://doi.org/10.1080/20002297.2016.1274582

Lebeer, S., et al. (2008). Genes and molecules of lactobacilli supporting probiotic action. Microbiology and Molecular Biology Reviews: MMBR, 72(4), 728.

Mack, D. R., et al. (2003). Extracellular MUC3 mucin secretion follows adherence of Lactobacillus strains to intestinal epithelial cells in vitro. Gut, 52(6), 827-833.

Martin, H., et al. (2020). Targeting adhesion in fungal pathogen Candida albicans. Future Medicinal Chemistry, 13(3), 313-334. https://doi.org/10.4155/fmc-2020-0052

Martins, N., et al. (2014). Candidiasis: predisposing factors, prevention, diagnosis and alternative treatment. Mycopathologia, 177(5), 223-240. https://doi.org/10.1007/s11046-014-9749-1

Megri, Y., et al. (2020). Candida tropicalis is the most prevalent yeast species causing candidemia in Algeria: the urgent need for antifungal stewardship and infection control measures. Antimicrobial Resistance and Infection Control, 9(1), 50. https://doi.org/10.1186/s13756-020-00710-z

Meurman, J. H., et al. (2018). Probiotics: evidence of oral health implications. Folia medica.

Miranda-Cadena, K., et al. (2018). Prevalence and antifungal susceptibility profiles of Candida glabrata, Candida parapsilosis and their close-related species in oral candidiasis. Archives of Oral Biology, 95, 100-107. https://doi.org/https://doi.org/10.1016/j.archoralbio.2018.07.017

Segers, M. E., et al. (2014). Towards a better understanding of Lactobacillus rhamnosus GG - host interactions. Microbial Cell Factories, 13(1), S7. https://doi.org/10.1186/1475-2859-13-S1-S7

Tournu, H., et al. (2012). Candida biofilms and the host: models and new concepts for eradication. Int J Microbiol, 2012, 845352. https://doi.org/10.1155/2012/845352

Tsui, C., et al. (2016). Pathogenesis of Candida albicans biofilm. Pathogens and Disease, 74(4). https://doi.org/10.1093/femspd/ftw018

Uppuluri, P., et al. (2010). Dispersion as an important step in the Candida albicans biofilm developmental cycle. PLoS Pathogen, 6(3), e1000828. https://doi.org/10.1371/journal.ppat.1000828

Yang, Y. (2003). Virulence factors of Candida species. Journal of Microbiology, immunology, and infection, 36 4, 223-228.

Zhang, C., et al. (2018). Stress influenced the aerotolerance of Lactobacillus rhamnosus hsryfm 1301. Biotechnology Letters, 40(4), 729-735. https://doi.org/10.1007/s10529-018-2523-6

Zuza-Alves, D. L., et al. (2017). An update on Candida tropicalis based on basic and clinical approaches. Frontiers in Microbiology, 8(1927). https://doi.org/10.3389/fmicb.2017.01927