Preliminary Assessment of the In Vitro Immunological Activity of Native Lactobacillus Strains from the Ecuadorian Amazon

Authors

DOI:

https://doi.org/10.70099/BJ/2024.01.03.20

Keywords:

cocoa, lactic acid bacteria, biological activity, isolation, cocoa fermentation, probiotics, cocoa pulp

Abstract

The isolation and characterization of new Lactobacillus strains from fermented traditional foods is a global trend as it enhances the potential for discovering novel probiotic foods. The fermented cocoa (Theobroma cacao) mucilage is an intriguing substrate for isolating lactic acid bacteria involved in fermentation. This study represents a preliminary investigation into the isolation, quantification, characterization, and immunological activity of Lactobacillus strains derived from cocoa beans sampled from two farmer associations (Kallari and Wiñak) in the Ecuadorian Amazon region. A mother culture was prepared using fermented cocoa pulp, cultivated on selective MRS media. After growth, the isolates were morphologically characterized. A significantly higher bacterial concentration was recorded in Kallari Association samples if compared with Wiñac. A total of 25 strains were isolated, eight of which were rod-shaped and positive to catalase tests and were characterized as Lactobacillus. In vitro, immunological activity was performed on differentiated THP-1 cell lines. Cells were treated with bacterial concentrates, and immunological activity was determined through interleukin-10 expression. Results indicated that W6 strain showed the highest immunological activity. These results indicated that Lactobacillus strains isolated from fermented cocoa pulp in the Ecuadorian Amazon show promise as a new source of probiotics.

References

1. Aparna Sudhakaran, V., Panwar, H., Chauhan, R., Duary, R.K., Rathore, R.K., Batish, V.K., Grover, S. Modulation of anti-inflammatory response in lipopolysaccharide stimulated human THP-1 cell line and mouse model at gene expression level with indigenous putative probiotic lactobacilli. Genes Nutr 2013, 8(6), 637-648. DOI: 10.1007/s12263-013-0347-5

2. Alard, J., Lehrter, V., Rhimi, M., Mangin, I., Peucelle, V., Abraham, A.L., Mariadassou, M., Maguin, E., Waligora-Dupriet, A.J., Pot, B., Wolowczuk, I., Grangette, C. Beneficial metabolic effects of selected probiotics on diet-induced obesity and insulin resistance in mice are associated with improvement of dysbiotic gut microbiota. Environ Microbiol 2016, 18(5), 1484–1497. DOI: 10.1111/1462-2920.13181

3. Hooper, L.V., Littman, D.R., Macpherson, A.J. Interactions between the microbiota and the immune system. Science 2012, 336(6086), 1268–1273. DOI: 10.1126/science.1223490

4. Kaji, R., Kiyoshima-Shibata, J., Nagaoka, M., Nanno, M., Shida, K. Bacterial teichoic acids reverse predominant IL-12 production induced by certain Lactobacillus strains into predominant IL-10 production via TLR2-dependent ERK activation in macrophages. J Immunol 2010, 184, 3505-3513. DOI: 10.4049/jimmunol.0901569

5. Smith, P., Garrett, W. The Gut Microbiota and Mucosal T Cells. Front. Microbiol. 2011, 2. https://doi.org/10.3389/fmicb.2011.00111

6. Duary, R.K., Bhausaheb, M.A., Batish, V.K., Grover, S. Anti-inflammatory and immunomodulatory efficacy of indigenous probiotic Lactobacillus plantarum Lp91 in colitis mouse model. Mol. Biol. Rep. 2012, 39, 4765–4775.

7. Swain, M.R., Anandharaj, M., Ray, R.C., Parveen Rani, R. Fermented Fruits and Vegetables of Asia: A Potential Source of Probiotics. Biotechnology Research International 2014, 250424. doi:10.1155/2014/250424

8. Granato, D., Branco, G.F., Nazzaro, F., Cruz, A.G., Faria, J.A.F. Functional Foods and Nondairy Probiotic Food Development: Trends, Concepts, and Products. Compr. Rev. Food Sci. Food Saf. 2010, 9, 292–302. doi: 10.1155/2014/250424

9. Saito, V.S.T., dos Santos, T.F., Vinderola, C.G., Romano, C., Nicoli, J.R., Araújo, L.S., Costa, M.M., Andrioli, J.L., Uetanabaro, A.P.T. Viability and resistance of lactobacilli isolated from cocoa fermentation to simulated gastrointestinal digestive steps in soy yogurt. J Food Sci 2014, 79(2), 208-213. DOI: 10.1111/1750-3841.12326

10. Qamsari, E.M., Kermanshahi, R.K., Erfan, M., Ghadam, P., Sardari, S., Eslami, N. Characteristics of surface layer proteins from two new and native strains of Lactobacillus brevis. Int J Biol Macromol. 2017, 95, 1004-1010. https://doi.org/10.1016/j.ijbiomac.2016.10.089

11. Aziz, G., Fakhar, H., Rahman, S.U., Tariq, M., Zaidi, A. An assessment of the aggregation and probiotic characteristics of Lactobacillus species isolated from native (desi) chicken gut. J Appl Poultry Res 2019, 28, 846-57.

12. Ordoñez-Araque, R.H., Landines-Vera, E.F., Urresto-Villegas, J.C., Caicedo-Jaramillo, C.F. Microorganisms during cocoa fermentation: Systematic review. Foods and Raw Materials, 2020, 8.1: 155-162. http://doi.org/10.21603/2308-4057-2020-1-155-162

13. Jadan, O., Cifuentes, M., Torres, B., Selesi, D., Veintimilla, D., Gunter, S. Influence of tree cover on diversity, carbon sequestration and productivity of cocoa systems in the Ecuadorian Amazon. Bois For Trop 2015, 325(3), 35–47. DOI:10.19182/bft2015.325.a31271

14. Crafack, M., Keul, H., Eskildsen, C.E., Petersen, M.A., Saerens, S., Blennow, A.., Skovmand-Larsen, M., Swiegers, J.H., Petersen, G.B., Heimdal, H., Nielsen, D.S. Impact of starter cultures and fermentation techniques on the volatile aroma and sensory profile of chocolate. Food Res Int 2014, 63, 306–316. https://doi.org/10.1016/j.foodres.2014.04.032

15. Kongor, J.E., Hinneh, M., Van de Walle, D., Afoakwa, E.O., Boeckx, P., Dewettinck, K. Factors influencing quality variation in cocoa (Theobroma cacao) bean flavour profile - A review. Food Res Int 2016, 82, 44–52. https://doi.org/10.1016/j.foodres.2016.01.012

16. Santos, T.T., Santos, Ornellas, R.M., Borges Arcucio, L., Messias Oliveira, M., Nicoli, R., Villela Dias, C., Trovatti Uetanabaro, A.P., Vinderola, G. Characterization of lactobacilli strains derived from cocoa fermentation in the south of Bahia for the development of probiotic cultures. LWT - Food Sci Technol 2016, 73, 259–266. https://doi.org/10.1016/j.lwt.2016.06.003

17. Visintin, S., Alessandria, V., Valente, A., Dolci, P., Cocolin, L. Molecular identification and physiological characterization of yeasts, lactic acid bacteria and acetic acid bacteria isolated from heap and box cocoa bean fermentations in West Africa. Int J Food Microbiol. 2016, 216, 69-78. DOI: 10.1016/j.ijfoodmicro.2015.09.004

18. Larrea, C., Kawachi, I. Does economic inequality affect child malnutrition? The case of Ecuador. Soc Sci Med 2005, 60,165-78. https://doi.org/10.1016/j.socscimed.2004.04.024

19. De Vuyst, L., Weckx, S. The cocoa bean fermentation process: from ecosystem analysis to starter culture development. J Appl Microbiol 2016, 121(1), 5-17. https://doi.org/10.1111/jam.13045

20. De Vuyst, L, Leroy, F. Functional role of yeasts, lactic acid bacteria and acetic acid bacteria in cocoa fermentation processes. FEMS Microbiology Reviews 2020, 44(4), 432-453. doi: 10.1093/femsre/fuaa014

21. Schwan, R, Wheals, A. The microbiology of cocoa fermentation and its role in chocolate quality. Critical Reviews in Food Science and Nutrition 2004, 44(4), 205-221. doi: 10.1080/10408690490464104

22. Camu, N., De Winter, T., Verbrugghe, K., Cleenwerck, I., Vandamme, P., Takrama, J.S., Vancanneyt, M., De Vuyst, L. Dynamics and biodiversity of populations of lactic acid bacteria and acetic acid bacteria involved in spontaneous heap fermentation of cocoa beans in Ghana. Appl Environ Microb 2007, 73(6), 1809-1824. DOI: 10.1128/AEM.02189-06

23. De Angelis, M., Gobbetti, M. Lactic Acid Bacteria. Lactobacillus spp.: General Characteristics. Reference Module in Food Science 2016. DOI:10.1016/B978-0-12-374407-4.00259-4

24. Morante-Carriel, L., Abasolo, F., Bastidas-Caldes, C., Paz, E.A., Huaquipán, R., Díaz, R., Valdes, M., Cancino, D., Sepúlveda, N., Quiñones, J. Isolation and Characterization of Lactic Acid Bacteria from Cocoa Mucilage and Meat: Exploring Their Potential as Biopreservatives for Beef. Microbiology Research, 2023, 14(3), 1150-1167. https://doi.org/10.3390/microbiolres14030077

25. Adikari, A., Priyashantha, H., Disanayaka, J., Jayatileka, D., Kodithuwakku, S., Jayatilake, J., Vidanarachchi, J. Isolation, identification and characterization of Lactobacillus species diversity from Meekiri: traditional fermented buffalo milk gels in Sri Lanka. Heliyon 2021, 7(10), https://doi.org/10.1016/j.heliyon.2021.e08136

26. Abid, S., Farid, A., Abid, R., Rehman, M., Alsanie, W.F., Alhomrani, M., Alamri, A.S., Asdaq, S.M.B., Hefft, D.I., Saqib, S., Muzammal, M, Morshedy, S.A., Alruways, M.W., Ghazanfar, S. Identification, Biochemical Characterization, and Safety Attributes of Locally Isolated Lactobacillus fermentum from Bubalus bubalis (Buffalo) Milk as a Probiotic. Microorganisms 2022, 10(5), https://doi.org/10.3390/microorganisms10050954

27. Taye, Y., Degu, T., Fesseha, H., Mathewos, M. Isolation and Identification of Lactic Acid Bacteria from Cow Milk and Milk Products. Scientific World Journal 2021, doi: 10.1155/2021/4697445

28. Genin, M., Clement, F., Fattaccioli, A., Raes, M., Michiels, C. M1 and M2 macrophages derived from THP-1 cells differentially modulate the response of cancer cells to etoposide. BMC Cancer 2015, Aug 8, 15, 577. DOI: 10.1186/s12885-015-1546-9

29. Voigt, J., Textoris-Taube, K., Wöstemeyer, J. pH-Dependency of the proteolytic formation of cocoa- and nutty-specific aroma precursors. Food Chem 2018, 255, 209-215. https://doi.org/10.1016/j.foodchem.2018.02.045

30. Ho, V.T.T., Zhao, J., Fleet, G. The effect of lactic acid bacteria on cocoa bean fermentation. Int J Food Microbiol 2015, 205, 54-67. https://doi.org/10.1016/j.ijfoodmicro.2015.03.031

31. Mudoor Sooresh, M., Willing, B.P., Bourrie, B.C.T. Opportunities and Challenges of Understanding Community Assembly in Spontaneous Food Fermentation. Foods 2023, 12, 673. https://doi.org/10.3390/foods12030673

32. Oliveira, J.S., Costa, K., Acurcio, L.B., Sandes, S.H.C., Cassali, G.D., Uetanabaro, A.P.T., Costa, A.M., Nicoli, J.R., Neumann, E., Porto, A.L.F. In vitro and in vivo evaluation of two potential probiotic lactobacilli isolated from cocoa fermentation (Theobroma cacao L.). J Func Foods 2018, 47, 184-191. https://doi.org/10.1016/j.jff.2018.05.055

33. Papalexandratou, Z., Kaasik, K., Villagra Kauffmann, L., Skorstengaard, A., Bouillon, G., Espensen, J.L., Hansen, L.H., Jakobsen, R.R., Blennow, A., Krych, L., Castro-Mejía, J.L., Nielsen, D.S. Linking cocoa varietals and microbial diversity of Nicaraguan fine cocoa bean fermentations and their impact on final cocoa quality appreciation. Int J Food Microbiol 2019, 304, 106-18. https://doi.org/10.1016/j.ijfoodmicro.2019.05.012

34. Idoui, T. Probiotic properties of Lactobacillus strains isolated from gizzard of local poultry. Iranian journal of microbiology 2014, 6, 120-26.

35. Da Cruz Pedroso Miguel, M.G., de Castro Reis, L.V., Efraim, P., Santos. C., Lima, N., Freitas Schwan, R. Cocoa fermentation: Microbial identification by MALDI-TOF MS, and sensory evaluation of produced chocolate. LWT 2017, 77, 362-69. https://doi.org/10.1016/j.lwt.2016.11.076

36. Kim, H.G., Gim, M.G., Kim, J.Y., Hwang, H.J., Ham, M.S., Lee, J.M., Hartung, T., Park, J.W., Han, S.H., Chung, D.K. Lipoteichoic acid from Lactobacillus plantarum elicits both the production of interleukin-23p19 and suppression of pathogen-mediated interleukin-10 in THP-1 cells. FEMS Immunol Med Microbiol 2007, 49, 205-14. DOI: 10.1111/j.1574-695X.2006.00175.x

37. Wang, Z., Xue, K., Bai, M., Deng, Z., Gan, J., Zhou, G., Qian, H., Bao, N., Zhao, J. Probiotics protect mice from CoCrMo particles-induced osteolysis. Int J Nanomed 2017, 12, 5387-5397. DOI: 10.2147/IJN.S130485

38. Lanaud, C., Vignes, H., Utge, J., Valette, G., Rhoné, B., Garcia Caputi, M., Angarita Nieto, N.S., Fouet, O., Gaikwad, N., Zarrillo, S., Powis, T.G., Cyphers, A., Valdez, F., Olivera Nunez, S.Q., Speller, C., Blake, M., Valdez, F.Jr., Raymond, S., Rowe, S.M., Duke, G.S., Romano, F.E., Loor Solórzano, R.G., Argout, X. A revisited history of cacao domestication in pre-Columbian times revealed by archaeogenomic approaches. Scientific Reports 2024, 14(1), https://doi.org/10.1038/s41598-024-53010-6

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Published

2024-09-15

How to Cite

Scalvenzi, L. (2024). Preliminary Assessment of the In Vitro Immunological Activity of Native Lactobacillus Strains from the Ecuadorian Amazon. BioNatura Journal: Ibero-American Journal of Biotechnology and Life Sciences, 1(3), 12. https://doi.org/10.70099/BJ/2024.01.03.20

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Vol. 1 No. 3 (2024)

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