Selection-Specific Bioactive Metabolites from Vernonia hymenolepis Wash Water Dictate Broiler Growth and Metabolic Efficiency in a Yeast-Dominant Fermentation System
The combined application of fermentation, phytobiotics, and endogenous probiotics offers a promising, sustainable alternative to conventional feed strategies, particularly for small-scale poultry production. This study evaluated the effects of fermenting a corn/soy diet using wash water (VhWW) derived from four distinct chemotypes of Vernonia hymenolepis (Green Bitter [GBVh, T4], Green Sweet [GSVh, T5], Purple Bitter [PBVh, T6], and Purple Sweet [PSVh, T7]) on broiler performance, nutrient utilization, and metabolism. A mandatory daily aeration step in the 72-hour staggered fermentation system created a novel Yeast-dominant, Lactic Acid Bacteria (LAB)-suppressed ecology across all VhWW treatments (≥8.00 log10CFU/mL Saccharomyces cerevisiae). This ecology conferred antimicrobial selectivity, achieving the elimination of E. coli in T4, T5, and T6. The PBVh selection (T6) yielded the highest Body Weight Gain (BWG: 2191.00 g, P < 0.001) and best Feed Conversion Ratio (FCR: 1.65, P = 0.053). However, its superior growth was decoupled from bulk nutrient absorption, as its corrected metabolizable dry matter (cMDM) was only intermediate. Conversely, the GBVh selection (T4) achieved the highest nutrient utilization (cMDM: 188.37 mg/g, P < 0.001) but one of the lowest BWGs, indicative of metabolic antagonism. This antagonism was supported by an adverse lipid profile (highest LDL and lowest HDL, P < 0.001). The PSVh selection (T7) induced metabolic stress (highest Relative Liver Weight, P < 0.001), poorest utilization, yet showed a repartitioning effect with the highest Relative Breast Yield (26.03%, P < 0.001). We conclude that the efficacy of VhWW fermentation is strain-dependent, with the PBVh selection maximizing performance by optimizing post-absorptive metabolic efficiency through bioactive metabolites derived from its unique phyto-microbial interaction.
References
[1]
Menchaca, A. (2021) Sustainable Food Production: The Contribution of Genome Editing in Livestock. Sustainability, 13, Article 6788. https://doi.org/10.3390/su13126788
[2]
da Costa Freitas, J. and Barbosa, M.J.B. (2025) Effective Strategies in Modern Livestock Management to Increase Productivity and Profitability. TheJournalofAcademicScience, 2, 1488-1495.
[3]
Coffey, D., Dawson, K., Ferket, P. and Connolly, A. (2016) Review of the Feed Industry from a Historical Perspective and Implications for Its Future. JournalofAppliedAnimalNutrition, 4, e3. https://doi.org/10.1017/jan.2015.11
[4]
Akintan, O., Gebremedhin, K.G. and Uyeh, D.D. (2024) Animal Feed Formulation—Connecting Technologies to Build a Resilient and Sustainable System. Animals, 14, Article 1497. https://doi.org/10.3390/ani14101497
[5]
Li, B., Boukhennou, A., Shao, J., Miao, L., Du, Y. and Chen, J. (2025) Application Status and Development Prospect of Fermented Ingredients in Aquaculture. AquacultureReports, 42, Article ID: 102842. https://doi.org/10.1016/j.aqrep.2025.102842
[6]
Chukwudi, P., Umeugokwe, P.I., Ikeh, N.E. and Amaefule, B.C. (2024) The Effects of Organic Acids on Broiler Chicken Nutrition: A Review. AnimalResearchandOneHealth, 3, 43-53. https://doi.org/10.1002/aro2.85
[7]
Choi, Y.H., Hosseindoust, A., Kim, M.J., Kim, K.Y., Lee, J.H., Kim, Y.H., etal. (2019) Additional Feeding during Late Gestation Improves Initial Litter Weight of Lactating Sows Exposed to High Ambient Temperature. RevistaBrasileiradeZootecnia, 48, e20180028. https://doi.org/10.1590/rbz4820180028
[8]
Sionek, B., Szydłowska, A., Küçükgöz, K. and Kołożyn-Krajewska, D. (2023) Traditional and New Microorganisms in Lactic Acid Fermentation of Food. Fermentation, 9, Article 1019. https://doi.org/10.3390/fermentation9121019
[9]
Ewane, D., Oben, B.O., Ndamukong, K.J.N., Etchu, K.A., Ehabe, E.E., Chah, J.M., etal. (2021) Toxicity of Auto-Detoxified Jatropha Curcas Linnaeus, 1753 Kernel Cake Mixtures with Bovine Blood (ADMJKC/bb) Using Brine Shrimp Artemia Salina Linnaeus, 1758. OnlineJournalofAnimalandFeedResearch, 11, 1-7. https://doi.org/10.51227/ojafr.2021.1
[10]
Ewane, D., Ndam, L.M., Keubiwou, B.R.F. and Boris, N.B. (2025) Evaluation of Vernonia Hymenolepis Wash Water as Phytogenic Prophylaxis for Broiler Chicken Production. SSRN Electronic Journal. https://doi.org/10.2139/ssrn.5481002
[11]
Ewane, D., Ndam, L.M., Agyingi, A.A., Ekeke, C.E.F., Ewane, E.E.E., Ewane, B.E., etal. (2025) Processing Vernonia amygdalina Delile to Reduce Competition for Food between Humans and Livestock. JournalofAgriculturalScience, 17, 46-62. https://doi.org/10.5539/jas.v17n12p46
[12]
Degu, S., Meresa, A., Animaw, Z., Jegnie, M., Asfaw, A. and Tegegn, G. (2024) Vernonia amygdalina: A Comprehensive Review of the Nutritional Makeup, Traditional Medicinal Use, and Pharmacology of Isolated Phytochemicals and Compounds. FrontiersinNaturalProducts, 3, Article 1347855. https://doi.org/10.3389/fntpr.2024.1347855
[13]
Afui, M.M., Kinge, R., Tonjock, and Ndam, L.M. (2008) Morphological Characterization of Four Selections of Vernoniahymenolepis A. Rich (Asteraceae). WorldJournalofAgriculturalSciences, 22, 220-223.
[14]
Yakubu, N. and Amuzat, A. (2012) Effect of Processing Methods on the Nutritional Contents of Bitter Leaf (Vernonia amygdalina). AmericanJournalofFoodandNutrition, 2, 26-30. https://doi.org/10.5251/abjna.2012.2.1.26.30
[15]
Abioye, V.F., Akande, E.A. and Aluko, B.O. (2014) Effects of Different Local Debittering Methods on Some Chemical Components and Antioxidants in Bitter Leaf (Vernonia amygdalina). InternationalJournalofResearchinChemistryandEnvironment, 4, 96-101.
[16]
Heres, L., Engel, B., Van Knapen, F., Wagenaar, J.A. and Urlings, B.A.P. (2003) Effect of Fermented Feed on the Susceptibility for Campylobacterjejuni Colonisation in Broiler Chickens with and without Concurrent Inoculation of Salmonellaenteritidis. InternationalJournalofFoodMicrobiology, 87, 75-86. https://doi.org/10.1016/s0168-1605(03)00055-2
[17]
Sibbald, I.R. (1979) A Bioassay for Available Amino Acids and True Metabolizable Energy in Feedingstuffs. PoultryScience, 58, 668-673. https://doi.org/10.3382/ps.0580668
[18]
Sugiharto, S. and Ranjitkar, S. (2019) Recent Advances in Fermented Feeds Towards Improved Broiler Chicken Performance, Gastrointestinal Tract Microecology and Immune Responses: A Review. AnimalNutrition, 5, 1-10. https://doi.org/10.1016/j.aninu.2018.11.001
[19]
Averina, O.V., Poluektova, E.U., Marsova, M.V. and Danilenko, V.N. (2021) Biomarkers and Utility of the Antioxidant Potential of Probiotic Lactobacilli and Bifido-bacteria as Representatives of the Human Gut Microbiota. Biomedicines, 9, Article 1340. https://doi.org/10.3390/biomedicines9101340
[20]
Klajn, V.M., Ames, C.W., da Cunha, K.F., Lorini, A., Hackbart, H.C.D.S., Filho, P.J.S., etal. (2021) Probiotic Fermented Oat Dairy Beverage: Viability of Lactobacilluscasei, Fatty Acid Profile, Phenolic Compound Content and Acceptability. JournalofFoodScienceandTechnology, 58, 3444-3452. https://doi.org/10.1007/s13197-021-04973-1
[21]
Merico, A., Sulo, P., Piškur, J. and Compagno, C. (2007) Fermentative Lifestyle in Yeasts Belonging to the Saccharomyces Complex. TheFEBSJournal, 274, 976-989. https://doi.org/10.1111/j.1742-4658.2007.05645.x
Matos, M.S., Anastácio, J.D. and Nunes dos Santos, C. (2021) Sesquiterpene Lactones: Promising Natural Compounds to Fight Inflammation. Pharmaceutics, 13, Article 991. https://doi.org/10.3390/pharmaceutics13070991
[24]
He, Y., Degraeve, P. and Oulahal, N. (2024) Bioprotective Yeasts: Potential to Limit Postharvest Spoilage and to Extend Shelf Life or Improve Microbial Safety of Processed Foods. Heliyon, 10, e24929. https://doi.org/10.1016/j.heliyon.2024.e24929
[25]
Spina, D. and Page, C.P. (2016) Xanthines and Phosphodiesterase Inhibitors. In: Page, C. and Barnes, P., Eds., Pharmacology and Therapeutics of Asthma and COPD Springer, 63-91. https://doi.org/10.1007/164_2016_71
[26]
Bernardes, A.C.F.P.F., Matosinhos, R.C., de Paula Michel Araújo, M.C., Barros, C.H., de Oliveira Aguiar Soares, R.D., Costa, D.C., etal. (2021) Sesquiterpene Lactones from Lychnophora Species: Antinociceptive, Anti-Inflammatory, and Antioxidant Pathways to Treat Acute Gout. JournalofEthnopharmacology, 269, Article ID: 113738. https://doi.org/10.1016/j.jep.2020.113738
[27]
Yu, X.H., Zheng, X.L. and Tang, C.K. (2015) Peroxisome Proliferator-Activated Receptor α in Lipid Metabolism and Atherosclerosis. Advances in Clinical Chemistry, 71, 171-203.
[28]
Guo, Y., Wang, B., Gao, H., Hua, R., Gao, L., He, C., etal. (2022) Peroxisome Proliferator-Activated Receptor-α: A Pivotal Regulator of the Gastrointestinal Tract. FrontiersinMolecularBiosciences, 9, Article 864039. https://doi.org/10.3389/fmolb.2022.864039
[29]
Sampath, V., Sureshkumar, S. and Kim, I.H. (2023) The Efficacy of Yeast Supplementation on Monogastric Animal Performance—A Short Review. Life, 13, Article 2037. https://doi.org/10.3390/life13102037
[30]
Svihus, B. (2011) The Gizzard: Function, Influence of Diet Structure and Effects on Nutrient Availability. World’sPoultryScienceJournal, 67, 207-224. https://doi.org/10.1017/s0043933911000249
[31]
Taha, T.M., Jalal, M.A., Zakaria, H.A. and Titi, H.H. (2025) Optimizing Hammer Mill Speed: Impact on Growth Performance, Pellet Quality, and Gizzard Gene Expression in Broilers. VeterinaryWorld, 18, 2542-2552. https://doi.org/10.14202/vetworld.2025.2542-2552
[32]
Arvind, A., Osganian, S.A., Cohen, D.E., etal. (2019) Lipid and Lipoprotein Metabolism in Liver Disease. In: Feingold, K.R., Ahmed, S.F., Anawalt, B., etal., Eds., Endotext, MDText.com, Inc. https://www.ncbi.nlm.nih.gov/books/NBK326742/
[33]
Pirahanchi, Y., Sinawe, H. and Dimri, M. (2023) Biochemistry, LDL Cholesterol. StatPearls Publishing.
[34]
Cattley, R.C. and Cullen, J.M. (2013) Liver and Gall Bladder. In: Haschek, V.R.F. and Rousseaux, C.G., Eds., HaschekandRousseaux’sHandbookofToxicologicPathology, Elsevier, 1509-1566. https://doi.org/10.1016/b978-0-12-415759-0.00045-5
[35]
Hu, Z., Xu, H., Lu, Y., He, Q., Yan, C., Zhao, X., etal. (2021) MUSTN1 Is an Indispensable Factor in the Proliferation, Differentiation and Apoptosis of Skeletal Muscle Satellite Cells in Chicken. ExperimentalCellResearch, 407, Article ID: 112833. https://doi.org/10.1016/j.yexcr.2021.112833
