Probiotic characterization of Limosilactobacillus fermentum BGHV110 strain and its influence on innate immune response in Caenorhabditis elegans
Abstract
Probiotic lactobacilli exhibit the potential to promote health benefits for the host. Thanks to its numerous beneficial effects on human health, Limosilactobacillus fermentum stood out as an excellent candidate for the development of commercial probiotic preparations aiming to prevent community-acquired infections. In this study, several in vitro tests, including biofilm formation assay, assessment of antibiotic susceptibility, survival in simulated gastrointestinal tract conditions and attachment to intestinal Caco-2 cells, were used to estimate the safety and probiotic potential of L. fermentum BGHV110 strain. Additionally, Caenorhabditis elegans was used as an in vivo model system for the evaluation of L. fermentum BGHV110 influence on the host’s innate immune response. The results revealed that L. fermentum BGHV110 strain showed an excellent capability to survive harsh conditions of the gut, to attach to intestinal Caco-2 cells and to stimulate conserved p38 MAPK immunity pathway and expression of the clc-1 claudin-like gene and antimicrobial peptides in C. elegans in order to enhance the immune response against pathogens. Finally, L. fermentum BGHV110 showed no virulence traits and susceptibility to tested antibiotics, confirming its safety status which enables it to be applied as a future probiotic.
References
Ragonnaud E, Biragyn A. Gut microbiota as the key controllers of "healthy" aging of elderly people. Immun Ageing. 2021;18(1):2.
Rajilić-Stojanović M. Function of the microbiota. Best Pract Res Clin Gastroenterol. 2013;27(1):5-16.
Sokovic Bajic S, Djokic J, Dinic M, Veljovic K, Golic N, Mihajlovic S, et al. GABA-producing natural dairy isolate from artisanal Zlatar cheese attenuates gut inflammation and strengthens gut epithelial barrier in vitro. Front Microbiol. 2019;10:527.
Dinić M, Pecikoza U, Djokić J, Stepanović-Petrović R, Milenković M, Stevanović M, et al. Exopolysaccharide produced by probiotic strain Lactobacillus paraplantarum BGCG11 reduces inflammatory hyperalgesia in rats. Front Pharmacol. 2018;9:1.
WHO-FAO. Probiotics in foods. Health and nutritional properties and guidelines for evaluation. FAO Food and Nutritional Paper, 2006; 8592-5-105513.
Naghmouchi K, Belguesmia Y, Bendali F, Spano G, Seal BS, Drider D. Lactobacillus fermentum: a bacterial species with potential for food preservation and biomedical applications. Crit Rev Food Sci Nutr. 2020;60(20):3387-3399.
Medzhitov R. Recognition of microorganisms and activation of the immune response. Nature. 2007;449(7164):819-26.
Dinić M, Jakovljević S, Đokić J, Popović N, Radojević D, Strahinić I, et al. Probiotic-mediated p38 MAPK immune signaling prolongs the survival of Caenorhabditis elegans exposed to pathogenic bacteria. Sci Rep. 2021;11(1):21258.
Santaolalla R, Fukata M, Abreu MT. Innate immunity in the small intestine. Curr Opin Gastroenterol. 2011;27(2):125-31.
Ermolaeva A, Schumacher B. Insights from the worm: the C. elegans model for innate immunity. Semin Immunol. 2014;26(4):303-9.
Asano A, Asano K, Sasaki H, Furuse M, Tsukita S. Claudins in Caenorhabditis elegans: their distribution and barrier function in the epithelium. Curr Biol. 2003;13(12):1042-6.
Roselli M, Schifano E, Guantario B, Zinno P, Uccelletti D, Devirgiliis C. Caenorhabditis elegans and probiotics interactions from a prolongevity perspective. Int J Mol Sci. 2019;20(20):5020.
Dinić M, Herholz M, Kačarević U, Radojević D, Novović K, Đokić J, et al. Host-commensal interaction promotes health and lifespan in Caenorhabditis elegans through the activation of HLH-30/TFEB-mediated autophagy. Aging (Albany NY). 2021;13(6):8040-8054.
Nakagawa H, Shiozaki T, Kobatake E, Hosoya T, Moriya T, Sakai F, et al. Effects and mechanisms of prolongevity induced by Lactobacillus gasseri SBT2055 in Caenorhabditis elegans. Aging Cell. 2016;15(2):227-36.
Kumar A, Joishy T, Das S, Kalita MC, Mukherjee AK, Khan MR. A Potential probiotic Lactobacillus plantarum JBC5 improves longevity and healthy aging by modulating antioxidative, innate immunity and serotonin signaling pathways in Caenorhabditis elegans. Antioxidants (Basel). 2022;11(2):268.
Dinić M, Lukić J, Djokić J, Milenković M, Strahinić I, Golić N, et al. Lactobacillus fermentum postbiotic-induced autophagy as potential approach for treatment of acetaminophen hepatotoxicity. Front Microbiol. 2017;8:594.
EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP), et al. Guidance on the characterisation of microorganisms used as feed additives or as production organisms. EFSA Journal. 2018;16(3):e05206.
Macovei L, Ghosh A, Thomas C, Hancock E, Mahmood S, Zurek L. Enterococcus faecalis with the gelatinase phenotype regulated by the fsr operon and with biofilm‐forming capacity are common in the agricultural environment. Environ Microbiol. 2009;11(6):1540-1547.
Sánchez B, Fernandez-Garcia M, Margolles A, de los Reyes-Gavilan G, Ruas-Madiedo P. Technological and probiotic selection criteria of a bile-adapted Bifidobacterium animalis subsp. lactis strain. Int Dairy J. 2010;20:7078-7085.
Anisimova E, Gorokhova I, Karimullina G, Yarullina D. Alarming antibiotic resistance of lactobacilli isolated from probiotic preparations and dietary supplements. Antibiotics (Basel). 2022;11(11):1557.
Campedelli I, Mathur H, Salvetti E, Clarke S, Rea C, Torriani S, et al. Genus-wide assessment of antibiotic resistance in Lactobacillus spp. Appl Environ Microbiol. 2019;85(1):e01738-18.
Cuevas-González F, Liceaga M, Aguilar-Toalá E. Postbiotics and paraprobiotics: From concepts to applications. Food Res Int. 2020;136:109502.
Gómez C, Ramiro M, Quecan X, de Melo Franco D. Use of potential probiotic lactic acid bacteria (LAB) biofilms for the control of Listeria monocytogenes, Salmonella Typhimurium, and Escherichia coli O157: H7 biofilms formation. Front Microbiol. 2016;7:863.
Kleerebezem M, Hols P, Bernard E, Rolain T, Zhou M, Siezen J, et al. The extracellular biology of the lactobacilli. FEMS Microbiol Rev. 2010;34(2):199-230.
Kaushik K, Kumar A, Duary K, Mohanty K, Grover S, Batish K. Functional and probiotic attributes of an indigenous isolate of Lactobacillus plantarum. PLoS One. 2009;4(12):e8099.
Tuomola M, Salminen J. Adhesion of some probiotic and dairy Lactobacillus strains to Caco-2 cell cultures. Int J Food Microbiol. 1998;41(1):45-51.
Panicker S, Ali A, Anand S, Panjagari R, Kumar S, Mohanty K, et al. Evaluation of some in vitro probiotic properties of Lactobacillus fermentum strains. J Food Sci Technol. 2018;55(7):2801-2807.
Jang J, Kim K, Han H, Lee K, Ko G. Lactobacillus fermentum species ameliorate dextran sulfate sodium-induced colitis by regulating the immune response and altering gut microbiota. Gut Microbes. 2019;10(6):696-711.
Lukic J, Strahinic I, Milenkovic M, Golic N, Kojic M, Topisirovic L, et al. Interaction of Lactobacillus fermentum BGHI14 with rat colonic mucosa: implications for colitis induction. Appl Environ Microbiol. 2013;79(18):5735-44.
Zarubin T, Han J. Activation and signaling of the p38 MAP kinase pathway. Cell Res. 2005;15(1):11-8.
Kim G, Ohta T, Takahashi T, Kushiro A, Nomoto K, Yokokura T, et al. Probiotic Lactobacillus casei activates innate immunity via NF-kappaB and p38 MAP kinase signaling pathways. Microbes Infect. 2006;8(4):994-1005.
Mørch M, Møller V, Hesselager O, Harders H, Kidmose L, Buhl T, et al. The TGF-β ligand DBL-1 is a key player in a multifaceted probiotic protection against MRSA in C. elegans. Sci Rep. 2021;11(1):10717.
