FERMENTED BUFFALO MILK (DADIH) POWDER AS PARABIOTICS WITH LOWERED WATER ACTIVITY TO PROTECT OXIDATIVE STRESS AND LIVER INJURY
Abstract
Dadih powder is dried fermented buffalo milk. This study evaluated the effectiveness of dadih powder enriched with modified cassava flour (MOCAF) on metabolic syndrome parameters in Sprague-Dawley rats. Formulations containing 10, 30, and 50% MOCAF were assessed for physicochemical properties and lactic acid bacteria (LAB) viability. The optimal formulation was determined using the deGarmo method, based on the proximate composition and total LAB, and identified the 10% MOCAF formulation as having the highest effectiveness score. This formulation was used in the intervention study. The rats were divided into four groups: normal diet (N), high-fat-high-sucrose diet (KN), and two treatment groups receiving dadih powder supplementation at 0.335 mg/g (D1) and 0.670 mg/g (D2) of body weight for eight weeks. The rats were then sacrificed, and blood and liver samples were collected for biochemical and histological analyses. Traditionally, fresh dadih is highly perishable, with a shelf life of approximately three days. Drying reduced the water activity from 0.70 to 0.66, thereby enhancing stability and prolonging its shelf life. The drying process effectively maintained the viability of LAB at level up to 102 cfu/g, thereby allowing the product to be classified as parabiotic. Dadih powder supplementation significantly reduced malondialdehyde levels, indicating antioxidative activity, although the reductions in glucose and lipid profiles were not significant. Histopathological analysis revealed severe hepatic steatosis in the KN group, whereas the D2 group exhibited near-normal liver structure. These findings indicate that dadih powder may serve as a parabiotic food to protect against oxidative stress and liver damage associated with metabolic syndrome.
References
Aalaei, K., Rayner, M., Tareke, E., & Sjöholm, I. (2016). Application of a dye-binding method for the determination of available lysine in skim milk powders. Food Chemistry, 196, 815–820. https://doi.org/10.1016/j.foodchem.2015.10.004
Ahmad, R., & Dalziel, J. E. (2020). G protein-coupled receptors in taste physiology and pharmacology. Frontiers in Pharmacology, 11(November), 587664. https://doi.org/10.3389/fphar.2020.587664
Almena, A., Goode, K. R., Bekalis, S., Fryer, P. J., & Lopez-Quiroga, E. (2019). Optimising food dehydration processes: energy-efficient drum-dryer operation. Energy Procedia, 161, 174–181. https://doi.org/10.1016/j.egypro.2019.02.078
AOAC. (2019). Official Methods of Analysis Of AOAC International (G. W. Latimer (ed.); 21st ed., Vol. 1). AOAC International.
Arnold, M., Rajagukguk, Y. V., & Gramza-Michałowska, A. (2021). Characterization of dadih: traditional fermented buffalo milk of minangkabau. Beverages, 7, 60. https://doi.org/10.3390/beverages7030060
Ary, E., Dadrasnia, A., Ameen, F., Alwakeel, S., & Ismail, S. (2021). Antimicrobial screening of lactic acid bacteria isolated from fermented milk buffalo (dadih). International Journal of Scientific and Research Publications (IJSRP), 11(4), 70–80. https://doi.org/10.29322/ijsrp.11.04.2021.p11209
Ayala, A., Muñoz, M. F., & Argüelles, S. (2014). Lipid peroxidation: Production , metabolism , and signaling mechanisms of malondialdehyde and 4-hydroxy-2-nonenal. Hindawi, 360438. https://doi.org/10.1155/2014/360438
Behbahani, B. A., Barzegar, H., & Taki, M. (2025). Isolation of Lacticaseibacillus rhamnosus SMHA30 from traditional yogurt: a probiotic with antimicrobial, anti-biofilm, and safety attributes. Applied Food Research, 5(1), 100882. https://doi.org/10.1016/j.afres.2025.100882
Bergwall, S., Johansson, A., Sonestedt, E., & Acosta, S. (2022). High versus low-added sugar consumption for the primary prevention of cardiovascular disease. Sex Differences in Cardiac Diseases: Pathophysiology, Presentation, Diagnosis and Management, 1(1), 33–54. https://doi.org/10.1002/14651858.CD013320.pub2
Burchfield, J. G., Kebede, M. A., Meoli, C. C., Stöckli, J., Whitworth, P. T., Wright, A. L., Hoffman, N. J., Minard, A. Y., Ma, X., Krycer, J. R., Nelson, M. E., Tan, S. X., Yau, B., Thomas, K. C., Wee, N. K. Y., Khor, E. C., Enriquez, R. F., Vissel, B., Biden, T. J., … Fazakerley, D. J. (2018). High dietary fat and sucrose results in an extensive and time-dependent deterioration in health of multiple physiological systems in mice. Journal of Biological Chemistry, 293(15), 5731–5745. https://doi.org/10.1074/jbc.RA117.000808
Buszewski, B., Rogowska, A., Pomastowski, P., Złoch, M., & Railean-Plugaru, V. (2017). Identification of microorganisms by modern analytical techniques. Journal of AOAC International, 100(6), 1607–1623. https://doi.org/10.5740/jaoacint.17-0207
Cheng, H., Yen, G., Huang, S., Chen, S., & Hsu, C. (2022). The next generation beneficial actions of novel probiotics as potential therapeutic targets and prediction tool for metabolic diseases. Journal of Food and Drug Analysis, 30(1). https://doi.org/10.38212/2224-6614.3396
Ciaula, A. Di, Garruti, G., Frühbeck, G., Angelis, M. De, Bari, O. de, Wang, Q.-H., D., Frank, L., & Piero, P. (2017). The role of diet in the pathogenesis of cholesterol gallstones. Physiology & Behavior, 176(3), 139–148. https://doi.org/10.1002/hep.30150.Ductular
Clarke, H. J., Mccarthy, W. P., Sullivan, M. G. O., Kerry, J. P., & Kilcawley, K. N. (2021). Oxidative quality of dairy powders: Influencing factors and analysis. Foods, 10, 2315. https://doi.org/10.3390/foods10102315
Diehl, K. L., Vorac, J., Hofmann, K., Meiser, P., Unterweger, I., Kuerschner, L., Weighardt, H., Förster, I., & Thiele, C. (2020). Kupffer cells sense free fatty acids and regulate hepatic lipid metabolism in high-fat diet and inflammation. Cells, 9(10). https://doi.org/10.3390/cells9102258
Farmanfarma, K. K., Kaykhaei, M. A., Adineh, H. A., Mohammadi, M., Dabiri, S., & Ansari-moghaddam, A. (2019). Prevalence of metabolic syndrome in Iran: A meta-analysis of 69 studies. Diabetes and Metabolic Syndrome: Clinical Research and Reviews, 13(1), 792–799. https://doi.org/10.1016/j.dsx.2018.11.055
Firdaus, J., Sulistyaningsih, E., & Subagio, A. (2018). Resistant starch modified cassava flour (MOCAF) improves insulin resistance. Asian Journal of Clinical Nutrition, 10(1), 32–36. https://doi.org/10.3923/ajcn.2018.32.36
Fusco, W., Lorenzo, M. B., Cintoni, M., Porcari, S., Rinninella, E., Kaitsas, F., Lener, E., Mele, M. C., Gasbarrini, A., Collado, M. C., Cammarota, G., & Ianiro, G. (2023). Short-chain fatty-scid-producing bacteria: key components of the human gut microbiota. Nutrients, 15, 2211. https://doi.org/10.3390/nu15092211
Gugliucci, A. (2023). Sugar and dyslipidemia: a double-hit, perfect storm. Journal of Clinical Medicine, 12(17). https://doi.org/10.3390/jcm12175660
Hasan, A. E. Z., Falah, S., Handharyani, E., & Dwicesaria, M. A. (2023). Transaminase enzyme activity and histopathology evaluation of rat’s liver induced by DMBA with temulawak extract (Curcuma xanthorrhiza). International Journal on Advanced Science, Engineering and Information Technology, 13(1), 267–275. https://doi.org/10.18517/ijaseit.13.1.17431
He, L., Xing, Y., Ren, X., Zheng, M., Yu, S., Wang, Y., Xiu, Z., & Dong, Y. (2022). Mulberry leaf extract improves metabolic syndrome by alleviating lipid accumulation in vitro and in vivo. Molecules, 27, 5111. https://doi.org/10.3390/MOLECULES27165111
Herlina, V. T., & Setiarto, R. H. B. (2024). From tradition to innovation: dadih, the minangkabau tribe’s traditional fermented buffalo milk from Indonesia. Journal of Ethnic Foods, 11(1). https://doi.org/10.1186/s42779-024-00234-6
Hossain, A., & Jayadeep, A. (2024). Drum drying of whole maize flour enhances bioaccessibility of zeaxanthin , α -tocopherol , phytosterols , phenolics and flavonoids. Journal of Cereal Science, 116, 103879. https://doi.org/10.1016/j.jcs.2024.103879
Jańczuk, A., Brodziak, A., Król, J., & Czernecki, T. (2023). Properties of yoghurt fortified in lactoferrin with effect of storage time. Animals, 13(10), 1–20. https://doi.org/10.3390/ani13101610
Jimenez, M., Flores-Andrade, E., Pascual-Pineda, L. A., & Beristain, C. I. (2015). Effect of water activity on the stability of Lactobacillus paracasei capsule. Food Science and Technology, 60, 346–351. https://doi.org/10.1016/j.lwt.2014.09.050
Juarez-enriquez, E., Olivas, G. I., Ortega-rivas, E., Zamudio-flores, P. B., & Perez-vega, S. (2019). Water activity, not moisture content, explains the influence of water on powder flowability. Food Science and Technology, 100, 35–39. https://doi.org/10.1016/j.lwt.2018.10.043
Kang, W., Pan, L., Peng, C., Dong, L., Cao, S., Cheng, H., Wang, Y., Zhang, C., Gu, R., Wang, J., & Zhou, H. (2020). Isolation and characterization of lactic acid bacteria from human milk. Journal of Dairy Science, 103(11), 9980–9991. https://doi.org/10.3168/jds.2020-18704
Kapar, F. S., & Ciftci, G. (2020). The effects of curcumin and Lactobacillus acidophilus on certain hormones and insulin resistance in rats with metabolic syndrome. Journal of Diabetes and Metabolic Disorders, 19(2), 907–914. https://doi.org/10.1007/s40200-020-00578-1
Kazancıgil, E., Demirci, T., Öztürk-Negiş, H. İ., & Akın, N. (2019). Isolation, technological characterization and in vitro probiotic evaluation of Lactococcus strains from traditional Turkish skin bag Tulum cheeses. Annals of Microbiology, 69(12), 1275–1287. https://doi.org/10.1007/s13213-019-01512-4
Khasanah, Y., Indrianingsih, A. W., Triwitono, P., & Murdiati, A. (2024). Production, biological activities and functional food of modified cassava flour (mocaf). Canrea Journal: Food Technology, Nutritions, and Culinary Journal, 7(2), 213–229. https://doi.org/10.20956/canrea.v7i2.1280
Kobi, J. B. B. S., Matias, A. M., Gasparini, P. V. F., Torezani-Sales, S., Madureira, A. R., da Silva, D. S., Correa, C. R., Garcia, J. L., Haese, D., Nogueira, B. V., de Assis, A. L. E. M., Lima-Leopoldo, A. P., & Leopoldo, A. S. (2023). High-fat, high-sucrose, and combined high-fat/high-sucrose diets effects in oxidative stress and inflammation in male rats under presence or absence of obesity. Physiological Reports, 11(7), 1–13. https://doi.org/10.14814/phy2.15635
Koirala, P., Malav, O. P., Rai, S., Palanisamy, G., Agrawal, A., Dhar, B. K., Bekhit, A. E. D. A., Deokar, G. S., & Nirmal, N. (2024). Impact of non-bovine milks and milk products on human gut microbiota: a perspective towards sustainable healthy food production. Trends in Food Science and Technology, 151(May), 104642. https://doi.org/10.1016/j.tifs.2024.104642
Labuza, T. P., & Altunakar, B. (2020). Water activity prediction and moisture sorption isotherms. In Water Activity in Foods: Fundamentals and Applications (2nd ed., pp. 161–205). John Wiley & Sons, Inc. https://doi.org/10.1002/9781118765982.ch7
Lasker, S., Rahman, M., Parvez, F., Zamila, M., & Miah, P. (2019). High-fat diet-induced metabolic syndrome and oxidative stress in obese rats are ameliorated by yogurt supplementation. Scientific Reports, 9, 20026. https://doi.org/10.1038/s41598-019-56538-0
Laurence, D. R., & Bacharach, A. L. (1964). Evaluation of drug activities: Pharmacometrics. Journal of Medicinal Chemistry, 1 (3). https://doi.org/10.1021/jm00321a067
Liu, Y., Wang, J., & Wu, C. (2022). Modulation of gut microbiota and immune system by probiotics, pre-biotics, and post-biotics. Frontiers in Nutrition, 8(January), 1–14. https://doi.org/10.3389/fnut.2021.634897
Machado, K. R. D., Tulini, F. L., Guimarães, J. das G. L., Moraes, I. C. F., Ditchfield, C., de Lima, Cesar Gonçalves Silva, V. L. dos S., & Favaro-Trindade, C. S. (2023). Production and evaluation of yogurt colored with anthocyanin-rich pigment prepared from jabuticaba (Myrciaria cauliflora Mart.) skin. Processes, 11, 526. https://doi.org/10.3390/pr11020526
Masenga, S. K., Kabwe, L. S., Chakulya, M., & Kirabo, A. (2023). Mechanisms of oxidative stress in metabolic syndrome. International Journal of Molecular Sciences, 24, 7898. https://doi.org/10.3390/ijms24097898
Mathur, H., Beresford, T. P., & Cotter, P. D. (2020). Health benefits of lactic acid bacteria (LAB) fermentates. Nutrients, 12(6), 1–16. https://doi.org/10.3390/nu12061679
Mohammadian, K., Fakhar, F., Keramat, S., & Stanek, A. (2024). The role of antioxidants in the treatment of metabolic dysfunction-associated fatty liver disease: a systematic review. Antioxidants, 13(7), 797. https://doi.org/10.3390/antiox13070797
Mohankumari, H. P., Naidu, K. A., Narasimhamurthy, K., & Vijayalakshmi, G. (2021). Bioactive pigments of monascus purpureus attributed to antioxidant, HMG-coA reductase inhibition and anti-atherogenic functions. Frontiers in Sustainable Food Systems, 5(February), 1–11. https://doi.org/10.3389/fsufs.2021.590427
Mulyadi, A. F., Febrianto, A., Kumalaningsih, S., & Aswari, A. W. (2012). Process engineering of drying milk powder with foam mat drying method, a study of the effect of the concentration and types of filler. Journal of Basic and Applied Research International, 2(4), 3588–3592.
Naibaho, J., Safithri, M., & Wijaya, C. H. (2019). Anti-hyperglycemic activity of encapsulated Java tea-based drink on malondialdehyde formation. Journal of Applied Pharmaceutical Science, 9(4), 88–95. https://doi.org/10.7324/JAPS.2019.90411
Nainggolan, E. A., Yudianto, D., & Sayekti, A. (2019). Effect of fermentation on physicochemical properties of fermented cassava flour. Journal of Physics: Conference Series, 1367(1), 012083. https://doi.org/10.1088/1742-6596/1367/1/012083
Nizar, J. M., Shepard, B. D., Vo, V. T., & Bhalla, V. (2018). Renal tubule insulin receptor modestly promotes elevated blood pressure and markedly stimulates glucose reabsorption. JCI Insight, 3(16), 1–12. https://doi.org/10.1172/jci.insight.95107
Peng, X., Xian, H., Ge, N., Hou, L., Tang, T., Xie, D., Gao, L., & Yue, J. (2024). Effect of probiotics on glycemic control and lipid profiles in patients with type 2 diabetes mellitus: a randomized, double blind, controlled trial. Frontiers in Endocrinology, 15(September), 1–12. https://doi.org/10.3389/fendo.2024.1440286
Pramana, A., Yudhistira, B., Jayalaksamana, M., Kurnia, D., Husnayain, N., & Pramitasari, R. (2025). Dadih, traditional fermented buffalo milk: a comprehensive review of the aspects of gastronomy, health benefits, and product development. Journal of Ethnic Foods, 12(1). https://doi.org/10.1186/s42779-024-00261-3
Ramos-García, M., Ble-Castillo, J. L., García-Vázquez, C., Tovilla-Zárate, C. A., Juárez-Rojop, I. E., Olvera-Hernández, V., Genis-Mendoza, A. D., Córdova-Uscanga, R., Álvarez-González, C. A., & Díaz-Zagoya, J. C. (2021). Effects of non-nutritive sweeteners on energy intake, body weight and postprandial glycemia in healthy and with altered glycemic response rats. Foods, 10(5). https://doi.org/10.3390/foods10050958
Reeves, P. G. (1997). Components of the AIN-93 diets as improvements in the AIN-76A diet. Experimental Biology, 127(March), 838–841.
Reis, S. A., Conceição, L. L., Rosa, D. D., Siqueira, N. P., & Peluzio, M. C. G. (2017). Mechanisms responsible for the hypocholesterolaemic effect of regular consumption of probiotics. Nutrition Research Reviews, 30(1), 36–49. https://doi.org/10.1017/S0954422416000226
Rizqiati, H., Nurwantoro, Susanti, S., Apriliani, I. N., & Prayoga, M. I. Y. (2024). The effect of skim milk powder on physical, chemical, and microbiology characteristics of goat milk kefir powder. Food Research, 8(3), 135–146. https://doi.org/10.26656/fr.2017.8(3).880
Saklayen, M. G. (2018). The global epidemic of the metabolic syndrome. Current Hypertension Reports, 20, 12. https://doi.org/10.1007/s11906-018-0812-z
Santoso, C. M. A., Bramantoro, T., Kardos, L., Szakács, D. F., & Nagy, A. (2022). Metabolic syndrome and periodontitis among adults: The 2018 Indonesia national health survey. Journal of Clinical Periodontology, 49(6), 562–572. https://doi.org/10.1111/jcpe.13622
Serrano, S., Grujović, M., Marković, K. G., Barreto-Crespo, M. T., & Semedo-Lemsaddek, T. (2025). From dormancy to eradication: Strategies for controlling bacterial persisters in food settings. Foods, 14(6), 1–30. https://doi.org/10.3390/foods14061075
Setyadjit, & Sukasih, E. (2015). Effect of addition of filler on the productuon of shallot (Allium cepa var. ascalonicum L.) powder with drum dryer. Procedia Food Science, 3, 396–408. https://doi.org/10.1016/j.profoo.2015.01.044
Shimizu, H., Masujima, Y., Ushiroda, C., Mizushima, R., Taira, S., Ohue-Kitano, R., & Kimura, I. (2019). Dietary short-chain fatty acid intake improves the hepatic metabolic condition via FFAR3. Scientific Reports, 9(1), 1–10. https://doi.org/10.1038/s41598-019-53242-x
Silva, V. R. R., Katashima, C. K., Lenhare, L., Silva, C. G. B., Morari, J., Camargo, R. L., Velloso, L. A., Saad, M. A., da Silva, A. S. R., Pauli, J. R., & Ropelle, E. R. (2017). Chronic exercise reduces hypothalamic transforming growth factor-β1 in middle-aged obese mice. Aging, 9(8), 1926–1940. https://doi.org/10.18632/aging.101281
Singh, sarojini, Sharma, R., Kumari, M., & Tiwari, S. (2019). Insulin receptors in the kidneys in health and disease. World Journal of Nephrology, 8(1), 11–22. https://doi.org/10.5527/wjn.v8.i1.11
Singh S, Sharma R, Kumari M, Tiwari S. Insulin receptors in the kidneys in health and disease. World J Nephrol 2019; 8(1): 11-22 [PMID: 30705868 DOI: 10.5527/wjn.v8.i1.11]
Sitanggang, A. B., Firdausi, N. Z., & Budijanto, S. (2021). Antioxidant activity of a mixture between water-soluble tempeh extract and whey powder that has undergone a Maillard reaction. Future of Food: Journal on Food, Agriculture and Society, 9(5). https://doi.org/10.17170/kobra-202110144897
Sornsenee, P., Chimplee, S., Saengsuwan, P., & Romyasamit, C. (2022). Characterization of probiotic properties and development of banana powder enriched with freeze-dried Lacticaseibacillus paracasei probiotics. Heliyon, 8(10), e11063. https://doi.org/10.1016/j.heliyon.2022.e11063
Subramaniyan, V., & Hanim, Y. U. (2025). Role of pancreatic lipase inhibition in obesity treatment: mechanisms and challenges towards current insights and future directions. International Journal of Obesity, June 2024, 492–506. https://doi.org/10.1038/s41366-025-01729-1
Tafu, N. N., & Jideani, V. A. (2022). Proximate, elemental, and functional properties of novel solid dispersions of moringa oleifera leaf powder. Molecules, 27(15). https://doi.org/10.3390/molecules27154935
Taha, S., El Abd, M., De Gobba, C., Abdel-Hamid, M., Khalil, E., & Hassan, D. (2017). Antioxidant and antibacterial activities of bioactive peptides in buffalo’s yoghurt fermented with different starter cultures. Food Science and Biotechnology, 26(5), 1325–1332. https://doi.org/10.1007/s10068-017-0160-9
Tamang, J. P., Shin, D. H., Jung, S. J., & Chae, S. W. (2016). Functional properties of microorganisms in fermented foods. Frontiers in Microbiology, 7(APR), 1–13. https://doi.org/10.3389/fmicb.2016.00578
Taranto, D., Ramirez, C. F. A., Vegna, S., de Groot, M. H. P., de Wit, N., Van Baalen, M., Klarenbeek, S., & Akkari, L. (2021). Multiparametric analyses of hepatocellular carcinoma somatic mouse models and their associated tumor microenvironment. Current Protocols, 1(6), 1–50. https://doi.org/10.1002/cpz1.147
Teo, P. S., Van Dam, R. M., Whitton, C., Tan, L. W. L., & Forde, C. G. (2021). Consumption of foods with higher energy intake rates is associated with greater energy intake, adiposity, and cardiovascular risk factors in adults. Journal of Nutrition, 151(2), 370–378. https://doi.org/10.1093/jn/nxaa344
Tkacz, K., Modzelewska-Kapituła, M., Więk, A., & Nogalski, Z. (2020). The applicability of total color difference ∆e for determining the blooming time in longissimus lumborum and semimembranosus muscles from holstein-friesian bulls at different ageing times. Applied Sciences, 10(22), 1–10. https://doi.org/10.3390/app10228215
Tsutsumi, R., Yamasaki, Y., Takeo, J., Miyahara, H., Sebe, M., Bando, M., Tanba, Y., Mishima, Y., Takeji, K., Ueshima, N., Kuroda, M., Masumoto, S., Harada, N., Fukuda, D., Yoshimoto, R., Tsutsumi, Y. M., Aihara, K. ichi, Sata, M., & Sakaue, H. (2021). Long-chain monounsaturated fatty acids improve endothelial function with altering microbial flora. Translational Research, 237, 16–30. https://doi.org/10.1016/j.trsl.2021.03.016
Valadares, L. T. de S., de Souza, L. S. B., Júnior, V. A. S., Bonomo, L. de F., de Macedo, L. R., & Silva, M. (2022). Prevalence of metabolic syndrome in Brazilian adults in the last 10 years: a systematic review and meta-analysis. BMC Public Health, 22, 327. https://doi.org/10.1186/s12889-022-12753-5
Vu, T. P., He, L., Mcclements, D. J., & Decker, E. A. (2020). Effects of water activity , sugars , and proteins on lipid oxidative stability of low moisture model crackers. Food Research International, 130(December 2019), 108844. https://doi.org/10.1016/j.foodres.2019.108844
Wan-Mohtar, W. A. A. Q. I., Ilham, Z., Jamaludin, A. A., David, W., & Mohd Zaini, N. A. (2022). Fermented foods as alternative functional foods during post-pandemic in Asia. Frontiers in Food Science and Technology, 2(February 2020), 1–7. https://doi.org/10.3389/frfst.2022.1047970
Yao, W., Liu, K., Liu, H., Jiang, Y., Wang, R., Wang, W., & Wang, T. (2021). A valuable product of microbial cell factories: microbial lipase. Frontiers in Microbiology, 12(September), 1–16. https://doi.org/10.3389/fmicb.2021.743377
Yüksel, A. N. (2021). Development of yoghurt powder using microwave-assisted foam-mat drying. Journal of Food Science and Technology, 58(7), 2834–2841. https://doi.org/10.1007/s13197-021-05035-2
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