Background: “Kossam”, a traditionally processed milk in Cameroon and sub-Saharan Africa, has been described as potential source of probiotic lactic acid bacteria (LAB). However, to our knowledge, no research work has been done on their immunomodulatory properties. Aim: This study was aimed at isolating and characterising lactic acid bacteria from “Kossam”, a traditionally fermented cow milk and investigating their immunomodulatory properties in rat model. Materials and Methods: Lactic acid bacteria were isolated from “Kossam” using pour plating technique. The strains exhibiting probiotic potential were selected and their identities were confirmed using the sequencing of 16S ribosomal RNA gene. Probiotic isolates were administered to rats for 2 consecutive days and a post ingestion period of 10 days was observed. Full blood count, differential blood count, total spleen protein and pro-inflammatory cytokines (TNF-α, IFN-g, IL-1-β and IL-6) concentrations were detected in the sera after 10 days. Results: A significant increase (p < 0.05) in the level of RBCs, lymphocytes, monocytes and total spleen proteins was observed in rats treated with the different isolates. However, a significant decrease (p < 0.05) in circulating WBCs and neutrophils count was observed in animals treated with the isolates A20, A27, A46, and A56 that had the highest effect (p < 0.05) on pro-inflammatory cytokines level in serum. These strains were identified using 16S rRNA gene sequence as strains of Lacticaseibacillusparacasei (A20), Limosilactobacillusfermentum (A27), Enterococcusfaecium (A46), Lactiplantibacillusplantarum (A56). Conclusion: These isolates showed immunomodulatory properties in rats and were able to prevent the incidence of diarrhoea associated with purgative components.
References
[1]
de Brito Alves, J.L., Monteiro, M., Sales-Campos, H. and de Souza, E.L. (2023) Editorial: Probiotics for Nutrition Research in Health and Disease. FrontiersinMicrobiology, 14, Article 1236442. https://doi.org/10.3389/fmicb.2023.1236442
[2]
Icer, M.A., Özbay, S., Ağagündüz, D., Kelle, B., Bartkiene, E., Rocha, J.M.F., et al. (2023) The Impacts of Acidophilic Lactic Acid Bacteria on Food and Human Health: A Review of the Current Knowledge. Foods, 12, Article 2965. https://doi.org/10.3390/foods12152965
[3]
Kioulaphides, S. and García, A.J. (2024) Encapsulation and Immune Protection for Type 1 Diabetes Cell Therapy. AdvancedDrugDeliveryReviews, 207, Article ID: 115205. https://doi.org/10.1016/j.addr.2024.115205
[4]
Ortuño-Sahagún, D., Zänker, K., Rawat, A.K.S., Kaveri, S.V. and Hegde, P. (2017) Natural Immunomodulators. JournalofImmunologyResearch, 2017, Article ID: 7529408. https://doi.org/10.1155/2017/7529408
[5]
Hegazy, R., Salama, A., Mansour, D. and Hassan, A. (2016) Renoprotective Effect of Lactoferrin against Chromium-Induced Acute Kidney Injury in Rats: Involvement of IL-18 and IGF-1 Inhibition. PLOSONE, 11, e0151486. https://doi.org/10.1371/journal.pone.0151486
[6]
Jacques, Y., Pierre, N.J., Vayaraï, M.A., Brice, E.J. and Dieudonné, A. (2022) Toxicological Risk Assessment of Dairy Products Marketed in the Markets of the City of Yaoundé, Cameroon. IssuesinBiologicalSciencesandPharmaceuticalResearch, 10, 7-11. https://doi.org/10.15739/ibspr.22.002
[7]
Fossi, B.T., Ekabe, D.E., Toukam, L.L., Tatsilong Pambou, H.O., Gagneux-Brunon, A., Nkenfou Nguefeu, C., et al. (2022) Probiotic Lactic Acid Bacteria Isolated from Traditional Cameroonian Palm Wine and Corn Beer Exhibiting Cholesterol Lowering Activity. Heliyon, 8, e11708. https://doi.org/10.1016/j.heliyon.2022.e11708
[8]
Sambrook, J., Fritsche, E. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual. 2nd Edition, Cold Harbor Laboratory Press.
[9]
Flore, T.N.E., François, Z.N. and Félicité, T.M. (2010) Immune System Stimulation in Rats by Lactobacillus sp. Isolates from Raffia Wine (Raphia Vinifera). CellularImmunology, 260, 63-65. https://doi.org/10.1016/j.cellimm.2009.09.002
[10]
Cai, Z., Guo, Y., Zheng, Q., Liu, Z., Zhong, G., Zeng, L., et al. (2024) Screening of a Potential Probiotic Lactiplantibacillus plantarum NUC08 and Its Synergistic Effects with Yogurt Starter. JournalofDairyScience, 107, 2760-2773. https://doi.org/10.3168/jds.2023-24113
[11]
Ngwa F.A., Bobga T.B., Toukam Tatsinkou L.L., Taiwe S.G. and Tatsinkou F.B. (2024) Ameliorative Effects of Probiotic Limosilactobacillus fermentum and Enterococcus lactis Isolated from Cameroonian Traditionally Processed Milk and Palm Wine against Chronic Constriction Injury Induced Neuropathic Pain in Mice. Journal of Ethernophamacology, 334, Article ID: 118560.
[12]
Hu, Y., Xie, Y., Su, Q., Fu, J., Chen, J. and Liu, Y. (2023) Probiotic and Safety Evaluation of Twelve Lactic Acid Bacteria as Future Probiotics. FoodbornePathogensandDisease, 20, 521-530. https://doi.org/10.1089/fpd.2023.0039
[13]
Wang, R., Li, N., Zheng, K. and Hao, J. (2018) Enhancing Acid Tolerance of the Probiotic Bacterium Lactobacillus acidophilus NCFM with Trehalose. FEMSMicrobiologyLetters, 365, fny217. https://doi.org/10.1093/femsle/fny217
[14]
Pennacchia, C., Ercolini, D., Blaiotta, G., Pepe, O., Mauriello, G. and Villani, F. (2004) Selection of Lactobacillus Strains from Fermented Sausages for Their Potential Use as Probiotics. MeatScience, 67, 309-317. https://doi.org/10.1016/j.meatsci.2003.11.003
[15]
Koponen, J., Laakso, K., Koskenniemi, K., Kankainen, M., Savijoki, K., Nyman, T.A., et al. (2012) Effect of Acid Stress on Protein Expression and Phosphorylation in Lactobacillus rhamnosus GG. JournalofProteomics, 75, 1357-1374. https://doi.org/10.1016/j.jprot.2011.11.009
[16]
Ortakci, F., Broadbent, J.R., McManus, W.R. and McMahon, D.J. (2012) Survival of Microencapsulated Probiotic Lactobacillus paracasei LBC-1e during Manufacture of Mozzarella Cheese and Simulated Gastric Digestion. JournalofDairyScience, 95, 6274-6281. https://doi.org/10.3168/jds.2012-5476
[17]
O'Shea, E.F., Cotter, P.D., Stanton, C., Ross, R.P. and Hill, C. (2012) Production of Bioactive Substances by Intestinal Bacteria as a Basis for Explaining Probiotic Mechanisms: Bacteriocins and Conjugated Linoleic Acid. InternationalJournalofFoodMicrobiology, 152, 189-205. https://doi.org/10.1016/j.ijfoodmicro.2011.05.025
[18]
Hou, C., Zeng, X., Yang, F., Liu, H. and Qiao, S. (2015) Study and Use of the Probiotic Lactobacillus reuteri in Pigs: A Review. JournalofAnimalScienceandBiotechnology, 6, Article No. 14. https://doi.org/10.1186/s40104-015-0014-3
[19]
Bao, Y., Zhang, Y., Zhang, Y., Liu, Y., Wang, S., Dong, X., et al. (2010) Screening of Potential Probiotic Properties of Lactobacillus fermentum Isolated from Traditional Dairy Products. FoodControl, 21, 695-701. https://doi.org/10.1016/j.foodcont.2009.10.010
[20]
Cebeci, A. and Gürakan, C. (2003) Properties of Potential Probiotic Lactobacillus plantarum Strains. FoodMicrobiology, 20, 511-518. https://doi.org/10.1016/s0740-0020(02)00174-0
[21]
Ruiz, L., Margolles, A. and Sánchez, B. (2013) Bile Resistance Mechanisms in Lactobacillus and Bifidobacterium. Frontiers in Microbiology, 4,Article 396. https://doi.org/10.3389/fmicb.2013.00396
[22]
Dabiré, Y., Somda, N.S., Somda, M.K., Compaoré, C.B., Mogmenga, I., Ezeogu, L.I., et al. (2022) Assessment of Probiotic and Technological Properties of Bacillus spp. Isolated from Burkinabe Soumbala. BMCMicrobiology, 22, Article No. 228. https://doi.org/10.1186/s12866-022-02642-7
[23]
Begley, M., Gahan, C.G.M. and Hill, C. (2005) The Interaction between Bacteria and Bile. FEMSMicrobiologyReviews, 29, 625-651. https://doi.org/10.1016/j.femsre.2004.09.003
[24]
Kingwatee, N., Apichartsrangkoon, A., Chaikham, P., Pankasemsuk, T. and Changrue, V. (2014) Survivability and Metabolic Activity of Lactobacillus casei 01 Incorporating Lychee Juice Plus Inulin under Simulated Gastrointestinal Environment. International Food Research Journal, 21, 83-89.
[25]
Jothi, V.V., Anandapandian, K. and Shankar, T. (2012) Bacteriocin Production by Pro-Biotic Bacteria from Curd and Its Field Application to Poultry. Archives of Applied Science Research, 4, 336-347.
[26]
Kusada, H., Morinaga, K. and Tamaki, H. (2021) Identification of Bile Salt Hydrolase and Bile Salt Resistance in a Probiotic Bacterium Lactobacillusgasseri JCM1131T. Microorganisms, 9, Article 1011. https://doi.org/10.3390/microorganisms9051011
[27]
Falor, A.E., Zobel, M., Kaji, A., Neville, A. and De Virgilio, C. (2012) Admission Variables Predictive of Gangrenous Cholecystitis. TheAmericanSurgeon, 78, 1075-1078. https://doi.org/10.1177/000313481207801014
[28]
Medzhitov, R. (2008) Origin and Physiological Roles of Inflammation. Nature, 454, 428-435. https://doi.org/10.1038/nature07201
[29]
Yu, Y., Yue, Z., Xu, M., Zhang, M., Shen, X., Ma, Z., et al. (2022) Macrophages Play a Key Role in Tissue Repair and Regeneration. PeerJ, 10, e14053. https://doi.org/10.7717/peerj.14053
[30]
Galdeano, C.M., Nunez, I.N., Carmuega, E., Moreno de LeBlanc, A.d. and Perdigon, G. (2015) Role of Probiotics and Functional Foods in Health: Gut Immune Stimulation by Two Probiotic Strains and a Potential Probiotic Yoghurt. Endocrine, Metabolic&ImmuneDisorders-DrugTargets, 15, 37-45. https://doi.org/10.2174/1871530314666141216121349
Abdi, M., Ghaleh, H.E.G. and Ranjbar, R. (2022) Lactobacilli and Bifidobacterium as Anti-Atherosclerotic Agents. IranianJournalofBasicMedicalSciences, 25, 934-946.
[33]
van Hemert, S., Meijerink, M., Molenaar, D., Bron, P.A., de Vos, P., Kleerebezem, M., et al. (2010) Identification of Lactobacillusplantarum Genes Modulating the Cytokine Response of Human Peripheral Blood Mononuclear Cells. BMCMicrobiology, 10, Article No. 293. https://doi.org/10.1186/1471-2180-10-293
[34]
Kaur, K., Nanut, M.P., Ko, M., Safaie, T., Kos, J. and Jewett, A. (2018) Natural Killer Cells Target and Differentiate Cancer Stem-Like Cells/Undifferentiated Tumors: Strategies to Optimize Their Growth and Expansion for Effective Cancer Immunotherapy. CurrentOpinioninImmunology, 51, 170-180. https://doi.org/10.1016/j.coi.2018.03.022
[35]
Ayob, N., Muhammad Nawawi, K.N., Mohamad Nor, M.H., Raja Ali, R.A., Ahmad, H.F., Oon, S.F., et al. (2023) The Effects of Probiotics on Small Intestinal Microbiota Composition, Inflammatory Cytokines and Intestinal Permeability in Patients with Non-Alcoholic Fatty Liver Disease. Biomedicines, 11, Article 640. https://doi.org/10.3390/biomedicines11020640
[36]
Maia, L.A., de Souza, J.R., da Silva, L.d.F.R., Magnani, M., de Souza, E.L. and de Brito Alves, J.L. (2024) Effects of Probiotics on Inflammatory Biomarkers and Its Associations with Cardiac Autonomic Function in Women with Arterial Hypertension: A Secondary Analysis of a Randomized Clinical Trial. ProbioticsandAntimicrobialProteins, 17, 1921-1928. https://doi.org/10.1007/s12602-024-10303-6
