No CrossRef data available.
Article contents
Nutritionally enriched whey-cereal-based fermented beverage with reduced faecal β-glucuronidase enzyme activity in a murine model
Published online by Cambridge University Press: 18 November 2025
Abstract
This research paper describes the effect of fermented substrates comprised of dairy by-products and underutilised cereals on murine faecal enzymes and faecal microbial profiles, and the development of the fermented substrate into a sour-spicy beverage for human consumption. A fermented substrate was made by using dairy by-products and underutilised cereals, whey and skim milk blend (60:40, v/v), germinated pearl millet flour and barley malt extract. The substrate was fermented with Lactobacillus acidophilus NCDC 13. The growth pattern of the organism in the composite substrate was satisfactorily described by a logistic-type equation. Faecal samples were obtained from 18 Swiss albino male mice that had been fed on either a control diet (n = 6), a diet based on an unfermented substrate, and a diet based on a fermented substrate (six in each group) and analysed. The fermented substrate caused a significant (P < 0.05) increment in faecal lactobacilli with a concomitant reduction in faecal coliform counts. Further, the fermented substrate caused a significant (P < 0.05) and sustainable decline in faecal enzyme β-glucuronidase activity in the mouse model, which is commonly considered a marker of colon cancer. The reductions in the numbers of coliform bacteria in faeces might explain the decline in faecal enzyme activity. Beta-glucuronidase is an enzyme produced by faecal bacteria that converts procarcinogens to potential carcinogens from available substrates. Bifidobacteria and lactobacilli generally have lower activities of these harmful enzymes, whereas β-glucuronidase is produced in high amounts by enterobacteria and clostridia spp. The fermented substrate was developed into a sour and spicy beverage for human consumption. The average TS, fat, protein, ash, starch and fibre contents of the beverage were 11%, 0.3%, 2%, 0.61%, 1% and 1.4%, respectively. The sensory score with an overall acceptability of 7.5, revealed that the product was sensory acceptable.
Information
- Type
- Research Article
- Information
- Copyright
- © The Author(s), 2025. Published by Cambridge University Press on behalf of Hannah Dairy Research Foundation.
References
AOAC (Association of Official Analytical Chemists) (1995) Official Methods of Analysis of AOAC International. Vol.11. 16th, Association of Official Analytical Chemists, Washington, DC: Virginia, USA.Google Scholar
Arora, T, Anastasovska, J, Gibson, G, Tuohy, K, Sharma, RK, Bell, J and Frost, G (2012) Effect of Lactobacillus acidophilus NCDC 13 supplementation on the progression of obesity in diet-induced obese mice. British Journal of Nutrition 108(8), 1382–1389.10.1017/S0007114511006957CrossRefGoogle ScholarPubMed
Chang, JH, Shim, YY, Cha, SK, Reaney, MJT and Chee, KM (2012) Effect of Lactobacillus acidophilus KFRI342 on the development of chemically induced precancerous growths in the rat colon. Journal of Medical Microbiology 61, 361–368.10.1099/jmm.0.035154-0CrossRefGoogle ScholarPubMed
Charalampopoulos, D, Pandiella, SS and Webb, C (2003) Evaluation of the effect of malt, wheat and barley extracts on the viability of potentially probiotic lactic acid bacteria under acidic conditions. International Journal of Food Microbiology 82(2), 133–141.10.1016/S0168-1605(02)00248-9CrossRefGoogle ScholarPubMed
Ganguly, S, Sabikhi, L and Singh, AK (2021) Evaluation of nutritional attributes of whey-cereal based probiotic beverage. LWT-Food Science and Technology 152, 112292. https://doi.org/10.1016/j.lwt.2021.112292.CrossRefGoogle Scholar
Ganguly, S, Sabikhi, L and Singh, AK (2022) Effect of probiotic fermentation on physico-chemical and nutritional parameters of milk-cereal based composite substrate. Journal of Food Science and Technology 59(8), 3073–3085.10.1007/s13197-021-05350-8CrossRefGoogle ScholarPubMed
De Filippis, F, Pasolli, E and Ercolini, D (2020) The food-gut axis: lactic acid bacteria and their link to food, the gut microbiome and human health. FEMS microbiology reviews 44(4), 454–489.10.1093/femsre/fuaa015CrossRefGoogle ScholarPubMed
Eileen, M, Melina, A, Gini, A, Lorenzoni, V, Cabasag, CJ, Mathieu, L and Freddie, B (2022) Global burden of colorectal cancer in 2020 and 2040: incidence and mortality estimates from GLOBOCAN. Gut 72(2), 338.Google Scholar
El-Aidie, SAM and Khalifa, GSA (2024) Innovative applications ofwhey protein for sustainable dairy industry: environmental and technological perspectives—Acomprehensive review. Comprehensive Reviews in Food Science and Food Safety 23, e13319. https://doi.org/10.1111/1541-4337.13319CrossRefGoogle ScholarPubMed
Emami, MH, Salehi, M, Hassanzadeh, KA, Mansourian, M, Mohammadzadeh, S and Maghool, F (2022) Calcium and dairy products in the chemoprevention of colorectal adenomas: a systematic review and meta-analysis. Critical Reviews in Food Science and Nutrition 62(26), 7168–7183.10.1080/10408398.2021.1911927CrossRefGoogle ScholarPubMed
Firmesse, O, Alvaro, E, Mogenet, A, Bresson, JL, Lemée, R, Le Ruyet, P, Bonhomme, C, Lambert, D, Andrieux, C, Doré, J and Corthier, G (2008) Fate and effects of Camembert cheese micro-organisms in the human colonic microbiota of healthy volunteers after regular Camembert consumption. International Journal of Food Microbiology 125, 176–181.10.1016/j.ijfoodmicro.2008.03.044CrossRefGoogle ScholarPubMed
Fu, J, Zheng, Y, Gao, Y and Xu, W (2022) Dietary fiber intake and gut microbiota in human health. Microorganisms 10(12), 2507.10.3390/microorganisms10122507CrossRefGoogle ScholarPubMed
Fujikawa, H, Kai, A and Morozumi, S (2004) A new logistic model for Escherichia coli growth at constant and dynamic temperatures. Food Microbiology 21, 501–509.10.1016/j.fm.2004.01.007CrossRefGoogle Scholar
Fukuda, M, Kanauchi, O, Araki, Y, Andoh, A, Mitsuyama, K, Takagi, K, Toyonaga, A, Sata, M and Fujiyama, Y (2002) Prebiotic treatment of experimental colitis with germinated barley foodstuff: a comparison with probiotic or antibiotic treatment. International Journal of Molecular Medicine 9, 65–70.Google ScholarPubMed
Ganguly, S and Sabikhi, L (2012) Fermentation dynamics of probiotic Lactobacillus acidophilus NCDC-13 in a composite dairy-cereal substrate. International Journal of Fermented Foods 1, 33–46.Google Scholar
Gill, SK, Rossi, M, Bajka, B and Whelan, K (2021) Dietary fibre in gastrointestinal health and disease. Nature Reviews Gastroenterology & Hepatology 18, 101–116.10.1038/s41575-020-00375-4CrossRefGoogle ScholarPubMed
Gomes, AMP and Malcata, FX (1999) Bifidobacterium spp. & Lactobacillus acidophilus: biological, biochemical, technological and therapeutical properties relevant for use as probiotics. Trends in Food Science and Technology 10, 139–157.10.1016/S0924-2244(99)00033-3CrossRefGoogle Scholar
Haughbty, GA, Maturin, LJ and Koening, EK (1993) In Marshall, RT (ed.), Microbiological Count Methods for Enumeration of Dairy Products. 16th Washington, DC: American Public Health Association (APHA). pp. 213–246.Google Scholar
Hendler, R and Zhang, Y (2018) Probiotics in the Treatment of Colorectal Cancer. Medicines 5(3), 101.10.3390/medicines5030101CrossRefGoogle ScholarPubMed
Hermes, RG, Molist, F, Ywazaki, M, Nofrarias, M, Gomez de Segura, A, Gasa, J and Perez, JF (2009) Effect of dietary level of protein and fibre on the productive performance and health status of piglets. Journal of Animal Science 87, 3569–3577.10.2527/jas.2008-1241CrossRefGoogle ScholarPubMed
Jood, S, Khetarpaul, N and Goyal, R (2012) Efficacy of barley based probiotic food mixture in treatment of pathogenic E.coli induced diarrhoea in mice. J Food Sci Technol 49, 200–206.10.1007/s13197-011-0270-yCrossRefGoogle ScholarPubMed
Kushal, R, Anand, SK and Chander, H (2005) Development of direct delivery system for coculture of L. acidophilus and B. bifidum based on micro-entrapment. Milchwissenschaft 60, 130–134.Google Scholar
Kushal, R, Anand, SK and Chander, H (2006) In vivo demonstration of enhanced probiotic effect of coimmobilized Lactobacillus acidophilus and Bifidobacterium bifidum. International Journal of Dairy Technology 59(4), 265–271.10.1111/j.1471-0307.2006.00283.xCrossRefGoogle Scholar
Le Roy, CI, Kurilshikov, A, Leeming, ER, Visconti, A, Bowyer, RCE, Menni, C, Fachi, M, Koutnikova, H, Veiga, P, Zhernakova, A and Derrien, M (2022)Yoghurt consumption is associated with changes in the composition of the human gut microbiome and metabolome. BMC Microbiology 22, 39. https://doi.org/10.1186/s12866-021-02364-2.CrossRefGoogle ScholarPubMed
Lee, D, Park, S, Jang, S, Baek, E, Kim, M, Huh, S, Choi, K, Chung, M, Kim, J, Lee, K and Ha, N (2011) The combination of mixed lactic acid bacteria and dietary fibre lowers serum cholesterol levels and faecal harmful enzyme activities in rats. Achieves of Pharmaceutical Research 34, 23–29.10.1007/s12272-011-0102-7CrossRefGoogle ScholarPubMed
Lee, J, Rheem, S, Yun, B, Ahn, Y, Joung, J, Lee, SJ, Oh, S, Chun, T, Rheem, I, Yea, HS, Lim, KS, Cha, JM and Kim, S (2013) Effects of probiotic yoghurt on symptoms and intestinal microbiota in patients with irritable bowel syndrome. International Journal of Dairy Technology 66, 243–255.10.1111/1471-0307.12028CrossRefGoogle Scholar
Leeuwendaal, NK, Stanton, C, O'Toole, PW and Beresford, TP (2022) Fermented foods, health and the gut microbiome. Nutrients 14(7), 1527. https://doi.org/10.3390/nu14071527CrossRefGoogle ScholarPubMed
Liang, Z, Song, X, Hu, J, Wu, R, Li, P, Dong, Z, Liang, L and Wang, J (2022) Fermented dairy food intake and risk of colorectal cancer: a systematic review and meta-analysis. Frontiers in Oncology 12, 812679. https://doi.org/10.3389/fonc.2022.812679CrossRefGoogle ScholarPubMed
Lisko, D, Johnston, G and Johnston, C (2017) Effects of dietary yogurt on the healthy human gastrointestinal (GI) microbiome. Microorganisms 5, 6.10.3390/microorganisms5010006CrossRefGoogle ScholarPubMed
Lukinac, J and Jukić, M (2022) Barley in the production of cereal-based products. Plants 11(24), 3519. https://doi.org/10.3390/plants11243519CrossRefGoogle ScholarPubMed
Lyra, A, Lahtinen, S and Ouwehand, AC (2012) Gastrointestinal benefits of probiotics – clinical evidence. Salminen, S, von Wright, A, Lahtinen, S and Ouwehand, A (eds.), Lactic Acid Bacteria: microbiological and Functional Aspects.4th. Boca Raton: CRC Press, pp. 509–523.Google Scholar
Marteau, P, Pochart, P, Flourie, B, Pellier, P, Santos, L, Desjeux, JF and Rambande, JC (1990) Effect of chronic ingestion of a fermented dairy product containing Lactobacillus acidophilus and Bifidobacterium bifidum on metabolic activities of the colonic flora in humans. American Journal of Clinical Nutrition 52(4), 685–688.10.1093/ajcn/52.4.685CrossRefGoogle ScholarPubMed
McMeekin, TA, Olley, J, Ratkowsky, DA and Ross, T (2002) Predictive microbiology: towards the interface and beyond. International Journal of Food Microbiology 73, 95–407.10.1016/S0168-1605(01)00663-8CrossRefGoogle ScholarPubMed
McNulty, NP, Yatsunenko, T, Hsiao, A, Faith, JJ, Muegge, BD, Goodman, AL, Henrissat, B, Oozeer, R, Cools-Portier, S, Gobert, G, Chervaux, C, Knights, D, Lozupone, CA, Knight, R, Duncan, AE, Bain, JR, Muehlbauer, MJ, Newgard, CB, Heath, AC and Gordon, JI (2011) The impact of a consortium of fermented milk strains on the gut microbiome of gnotobiotic mice and monozygotic twins. Science Transitional Medicine 3(106), 106ra–106.Google ScholarPubMed
Meena, KK, Meena, S, Joshi, M and Dhotre, AV (2024) Nutritional and functional exploration of pearl millet and its processing and utilization: an overview. Food and Humanity 3, 100334. https://doi.org/10.1016/j.foohum.2024.100334CrossRefGoogle Scholar
Mohammed, SG, Sahi, AA, Ameer, NA and Fusheng, C (2011) Prebiotic and synbiotic effects of Lactobacillus rhamnosus isolated from Iraq on intestinal tract microflora in mice. Pakistan Journal of Nutrition 10(5), 433–442.Google Scholar
Nielsen, ES, Garnås, E, Jensen, KJ, Hansen, LH, Olsen, PS, Ritz, C, Krych, L and Nielsen, DS (2018) Lacto-fermented sauerkraut improves symptoms in IBS patients independent of product pasteurisation—A pilot study. Food Funct 9, 5323–5335.10.1039/C8FO00968FCrossRefGoogle ScholarPubMed
Piccioni, A, Marcello, C, Marcello, C, Veronica, O, Annunziata, C, Antonio, G, Francesco, F and Giuseppe, M (2023) How do diet patterns, single foods, prebiotics and probiotics impact gut microbiota? Microbiology Research 14(1), 390–408.10.3390/microbiolres14010030CrossRefGoogle Scholar
Pillai, AT, Morya, S and Kasankala, LM (2024) Emerging trends in bioavailability and pharma‐nutraceutical potential of whey bioactives. Journal of Nutrition and Metabolism 2024(1), 8455666.10.1155/2024/8455666CrossRefGoogle ScholarPubMed
Pires, AF, Marnotes, NG, Rubio, OD, Garcia, AC and Pereira, CD (2021) Dairy by-products: a review on the valorization of whey and second cheese whey. Foods 10(5), 1067. https://doi.org/10.3390/foods10051067CrossRefGoogle ScholarPubMed
Priya, VRK, Lakhawat, S, Yadav, VK, Gacem, A, Abbas, M, Yadav, KK, Park, HK, Jeon, BH and Mishra, S (2023) Millets: sustainable treasure house of bioactive components. International Journal of Food Properties 26(1), 1822–1840.10.1080/10942912.2023.2236317CrossRefGoogle Scholar
Raghuramulu, N, Nair, KM and Kalyanasundaram, S (2003) A Manual of Laboratory Techniques. Hyderabad, India: NIN Press, National Institute of Nutrition, Jamai-Osmania.Google Scholar
Raj, R, Shams, R, Pandey, VK, Dash, KK, Singh, P and Bashir, O (2023) Barley phytochemicals and health promoting benefits: a comprehensive review. Journal of Agriculture and Food Research 14, 100677.10.1016/j.jafr.2023.100677CrossRefGoogle Scholar
Rocha-Mendoza, D, Kosmerl, E, Krentz, A, Zhang, L, Badiger, S, Miyagusuku-Cruzado, G, Mayta-Apaza, A, Giusti, M, Jiménez-Flores, R and García-Cano, I (2021) Invited review: acid whey trends and health benefits. Journal of Dairy Science 104(2), 1262–1275.10.3168/jds.2020-19038CrossRefGoogle ScholarPubMed
Sabikhi, L and Mathur, BN (2004) Intestinal microflora and carcinogenesis as influenced by dietary bifidobacteria delivered through Edam cheese to rats. Indian Journal of Dairy Science 57(6), 380–384.Google Scholar
Satyavathi, CT, Ambawat, S, Khandelwal, V and Srivastava, RK (2021) Pearl millet: a climate-resilient nutricereal for mitigating hidden hunger and provide nutritional security. Frontiers in Plant Science 12, 659938. https://doi.org/10.3389/fpls.2021.659938CrossRefGoogle ScholarPubMed
Scheppach, W, Luehrs, H and Menzel, T (2001) Beneficial health effects of low digestibility carbohydrate consumption. British Journal of Nutrition 85(1), S23–S30.10.1079/BJN2000259CrossRefGoogle ScholarPubMed
Shrestha, N, Hu, H, Shrestha, K and Doust, AN (2023) Pearl millet response to drought: a review. Frontiers in Plant Science 14, 1059574.10.3389/fpls.2023.1059574CrossRefGoogle ScholarPubMed
Sindhu, SC and Khetarpaul, N (2003) Fermentation with one step single and sequential cultures of yeast and lactobacilli: effect on antinutrients and digestibilities (in vitro) of starch and protein in an indigenously developed food mixture. Plant Foods for Human Nutrition 58(3), 1–10.10.1023/B:QUAL.0000040326.72490.83CrossRefGoogle Scholar
Singh, KP, Mishra, HN and Saha, S (2011) Sorption isotherms of Barnyard millet grain and Kernel. Food Bioprocess Technology 4(5), 788–796.10.1007/s11947-009-0195-xCrossRefGoogle Scholar
Stone, H, Sidel, J, Oliver, S, Woosley, A and Singleton, RC (1974) Sensory evaluation by quantitative descriptive analysis. Food Technology 28, 24–34.Google Scholar
Sung, H, Ferlay, J, Siegel, RL, Laversanne, M, Soerjomataram, I, Jemal, A and Bray, F (2021) Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer J Clin 71, 209–249.Google ScholarPubMed
Tanwar, R, Panghal, A, Kumari, A and Chhikara, N (2025) Nutritional, phytochemical and functional potential of pearl millet: a Review. Chemistry & Biodiversity, 22(7), e202402437.10.1002/cbdv.202402437CrossRefGoogle ScholarPubMed
Taylor, BC, Lejzerowicz, F, Poirel, M, Shaffer, JP, Jiang, L, Aksenov, A, Litwin, N, Humphrey, G, Martino, C, Miller-Montgomery, S and Dorrestein, PC (2020) Consumption of fermented foods is associated with systematic differences in the gut microbiome and metabolome. mSystems 5, 901–920.Google ScholarPubMed
Tripathy, A, Dash, J, Kancharla, S, Kolli, P, Mahajan, D, Senapati, S and Jena, MK (2021) Probiotics: a promising candidate for management of colorectal cancer. Cancers 13(13), 3178.10.3390/cancers13133178CrossRefGoogle ScholarPubMed
Veiga, P, Pons, N, Agrawal, A, Oozeer, R, Guyonnet, D, Brazeilles, R, Faurie, JM, van Hylckama Vlieg, JE, Houghton, LA, Whorwell, PJ and Ehrlich, SD (2014) Changes of the human gut microbiome induced by a fermented milk product. Scientific Reports 4, 6328.10.1038/srep06328CrossRefGoogle ScholarPubMed
Verma, A and Shukla, G (2013) Probiotics Lactobacillus rhamnosus GG, Lactobacillus acidophilus suppresses DMH-induced rocarcinogenic faecal enzymes and preneoplastic aberrant crypt foci in early colon carcinogenesis in sprague dawley rats. Nutrition and Cancer 65, 84–91.10.1080/01635581.2013.741746CrossRefGoogle ScholarPubMed
Verma, A and Shukla, G (2015) Modulation of apoptosis and immune response by synbiotic in experimental colorectal cancer. International Journal of Pharmaceurical Biological Science 6, 529–543.Google Scholar
Videla, S, Vilaseca, J, Antolin, M, Garcia-Lafuente, A, Guarner, F, Crespo, E, Casalots, J, Salas, A and Malagelada, JR (2001) Dietary inulin improves distal colitis induced by dextran sodium sulfate in the rat. American Journal of Gastroenterology 96, 1486–1493.10.1111/j.1572-0241.2001.03802.xCrossRefGoogle ScholarPubMed
Wollowski, I, Rechkemmer, G and Pool-Zobel, BL (2001) Protective role of probiotics and prebiotics in colon cancer. American Journal of Clinical Nutrition 73, 451S–455S.10.1093/ajcn/73.2.451sCrossRefGoogle ScholarPubMed
Yılmaz, İ, Enver Dolar, M and Özpınar, H (2019) Effect of administering kefir on the changes in faecal microbiota and symptoms of inflammatory bowel disease: a randomized controlled trial. Turkish Journal of Gastroenterology 30, 242–253.10.5152/tjg.2018.18227CrossRefGoogle ScholarPubMed
Yousaf, L, Hou, D, Liaqat, H and Shen, Q (2021) Millet: a review of its nutritional and functional changes during processing. Food Research International 142(3), 110197. https://doi.org/10.1016/j.foodres.2021.110197CrossRefGoogle Scholar
Zhang, J, Huan, D, Juan, B, Xinghua, Z, Yansheng, Z, Ying, Z, David, JM, Xiang, X and Quancai, S (2023) Health-promoting properties of barley: a review of nutrient and nutraceutical composition, functionality, bioprocessing, and health benefits. Critical Reviews in Food Science and Nutrition 63(9), 1155–1169.10.1080/10408398.2021.1972926CrossRefGoogle Scholar