Co-ensiling corn silage with Pleurotus-based spent mushroom substrate and oilseeds: Impacts on rumen fermentation, digestibility, and methane emissions in a RUSITEC system

Chika C. Anotaenwere , Omoanghe S. Isikhuemhen , Peter A. Dele , Ahmed E. Kholif , Michael Wuaku , Oludotun O. Adelusi , Deborah O. Okedoyin , Joel O. Alabi , Kelechi A. Ike , John O. Adebayo , Kiran Subedi , Uchenna Y. Anele

AIMS Microbiology ›› 2026, Vol. 12 ›› Issue (2) : 192 -210.

PDF (492KB)
AIMS Microbiology ›› 2026, Vol. 12 ›› Issue (2) :192 -210. DOI: 10.3934/microbiol.2026008
Research article
research-article
Co-ensiling corn silage with Pleurotus-based spent mushroom substrate and oilseeds: Impacts on rumen fermentation, digestibility, and methane emissions in a RUSITEC system
Author information +
History +
PDF (492KB)

Abstract

The present study examined the effects of partially replacing corn silage (CS) with Pleurotus-based spent mushroom substrate (SMS) and oilseeds (OS) on rumen fermentation characteristics, nutrient digestibility, volatile fatty acid (VFA) profiles, and greenhouse gas emissions using a rumen simulation technique (RUSITEC). Four silages were evaluated: A control diet containing 100% CS and three co-ensiled treatments in which CS was partially replaced with Pleurotus-based SMS and OS. The oilseed fraction consisted of equal proportions of soybean and canola seeds. The treatments were: CS90 (90% CS + 5% SMS + 2.5% soybean seed + 2.5% canola seed), CS80 (80% CS + 10% SMS + 5% soybean seed + 5% canola seed), and CS60 (60% CS + 20% SMS + 10% soybean seed + 10% canola seed). Dry matter (DM), organic matter, and crude protein contents were not affected by treatment, whereas ash content increased (P < 0.001) with increasing SMS-OS inclusion. Increasing SMS-OS inclusion significantly reduced non-fiber carbohydrates, hemicellulose and cellulose, while ether extract, neutral detergent fiber, acid detergent fiber, and acid detergent lignin concentrations were increased (P < 0.05). Ruminal fermentation pH, effluent volume, and total gas production showed only numerical differences or trends. DM digestibility increased markedly with SMS-OS inclusion (P < 0.001), rising from 38.7% in the control to 53.0, 49.9, and 52.5% in CS90, CS80, and CS60, respectively, representing improvements of approximately 29–37%. In CS90, organic matter digestibility remained high (P < 0.001) and fiber digestibility was largely the same. Moreover, CS90 had the highest total VFA and acetate, whereas CS60 had the highest propionate proportion (P < 0.05). All SMS-OS diets substantially reduced methane production (P = 0.003) and carbon dioxide emissions (P = 0.014) relative to the control. Methane output declined by approximately 60, 45, and 35% in CS90, CS80, and CS60, respectively (34.2–55.6 vs. 85.7 mg/g DM), while carbon dioxide emissions were reduced by about 49, 26, and 17% (134.7–218.4 vs. 264.0 mg/g DM). In conclusion, co-ensiling corn silage with SMS and oilseeds improved DM utilization while influencing methane production, with reductions observed in all the treatments compared with the control without adversely affecting rumen fermentation stability or fiber digestion under RUSITEC conditions.

Keywords

spent mushroom substrate / oilseeds / methane mitigation / corn silage / greenhouse gases / fiber digestibility

Cite this article

Download citation ▾
Chika C. Anotaenwere, Omoanghe S. Isikhuemhen, Peter A. Dele, Ahmed E. Kholif, Michael Wuaku, Oludotun O. Adelusi, Deborah O. Okedoyin, Joel O. Alabi, Kelechi A. Ike, John O. Adebayo, Kiran Subedi, Uchenna Y. Anele. Co-ensiling corn silage with Pleurotus-based spent mushroom substrate and oilseeds: Impacts on rumen fermentation, digestibility, and methane emissions in a RUSITEC system. AIMS Microbiology, 2026, 12 (2) : 192-210 DOI:10.3934/microbiol.2026008

登录浏览全文

4963

注册一个新账户 忘记密码

References

[1]

Daszkiewicz T (2022) Food production in the context of global developmental challenges. Agriculture 12: 832. https://doi.org/10.3390/agriculture12060832

[2]

Faizan M (2024) Enteric methane production in ruminants: Its effect on global warming and mitigation strategies-a review. Pak J Sci 76: 16-38. https://doi.org/10.57041/pjs.v76i01.1084

[3]

Hristov AN (2024) Invited review: Advances in nutrition and feed additives to mitigate enteric methane emissions. J Dairy Sci 107: 4129-4146. https://doi.org/10.3168/jds.2023-24440

[4]

Oliveira JS de, Oliveira JP de, Oliveira CP de, et al. (2025) Circular economy as a tool for the valorization of agro-industrial residues and the development of biodegradable and sustainable packaging: A bibliometric review from 2015 to 2024. Clean Circ Bioecon 12: 100188. https://doi.org/10.1016/j.clcb.2025.100188

[5]

Martín C, Xiong S, Zervakis GI (2025) Exploiting the biorefinery potential of spent mushroom substrate: The time to do it is now. Molecules 30: 4518. https://doi.org/10.3390/molecules30234518

[6]

Martín C, Zervakis GI, Xiong S, et al. (2023) Spent substrate from mushroom cultivation: Exploitation potential toward various applications and value-added products. Bioengineered 14: 2252138. https://doi.org/10.1080/21655979.2023.2252136

[7]

Anotaenwere CC, Isikhuemhen OS, Dele PA, et al. (2025) Enhanced ruminant nutrition and greenhouse gas abatement using co-ensiled corn silage, oilseeds, and spent mushroom substrate. Ger J Vet Res 5: 1-17. https://doi.org/10.51585/gjvr.2025.3.0144

[8]

Wuaku M, Isikhuemhen OS, Dele PA, et al. (2025) Solid state fermentation: A strategy for wheat bran supplemented corn stover valorization with Pleurotus species. AIMS Microbiol 11: 206-227. https://doi.org/10.3934/microbiol.2025011

[9]

Nie H, Wang Z, You J, et al. (2020) Comparison of in vitro digestibility and chemical composition among four crop straws treated by Pleurotus ostreatus. Asian Australas J Anim Sci 33: 24-34. https://doi.org/10.5713/ajas.18.0023

[10]

Koutrotsios G, Mountzouris KC, Chatzipavlidis I, et al. (2014) Bioconversion of lignocellulosic residues by Agrocybe cylindracea and Pleurotus ostreatus mushroom fungi-Assessment of their effect on the final product and spent substrate properties. Food Chem 161: 127-135. https://doi.org/10.1016/j.foodchem.2014.03.121

[11]

Anotaenwere CC, Isikhuemhen OS, Dele PA, et al. (2024) Ensiled Pleurotus ostreatus based spent mushroom substrate from corn: In vitro gas production, greenhouse gas emissions, nutrient degradation, and ruminal fermentation characteristics. AIMS Microbiol 11: 1-21. https://doi.org/10.3934/microbiol.2025001

[12]

Rangubhet KT, Mangwe MC, Mlambo V, et al. (2017) Enteric methane emissions and protozoa populations in Holstein steers fed spent mushroom (Flammulina velutipes) substrate silage-based diets. Anim Feed Sci Technol 234: 78-87. https://doi.org/10.1016/j.anifeedsci.2017.06.005

[13]

Xie X, Cao Y, Li Q, et al. (2025) Mitigating enteric methane emissions: An overview of methanogenesis, inhibitors and future prospects. Anim Nutr 21: 84-96. https://doi.org/10.1016/j.aninu.2025.02.001

[14]

Beauchemin KA, Ungerfeld EM, Eckard RJ, et al. (2020) Review: Fifty years of research on rumen methanogenesis: Lessons learned and future challenges for mitigation. Animal 14: s2-s16. https://doi.org/10.1017/S1751731119003100

[15]

Pepeta BN, Hassen A, Tesfamariam EH (2024) Quantifying the impact of different dietary rumen modulating strategies on enteric methane emission and productivity in ruminant livestock: A meta-analysis. Animals 14: 763. https://doi.org/10.3390/ani14050763

[16]

AOAC (2019) Official Methods of Analysis of AOAC International, Washington DC: Oxford University Press.

[17]

Van Soest PJ, Robertson JB, Lewis BA (1991) Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J Dairy Sci 74: 3583-3597. https://doi.org/10.3168/jds.S0022-0302(91)78551-2

[18]

Olagunju LK, Isikhuemhen OS, Dele PA, et al. (2023) Pleurotus ostreatus can significantly improve the nutritive value of lignocellulosic crop residues. Agriculture 13: 1161. https://doi.org/10.3390/agriculture13061161

[19]

Lay JJ, Li YY, Noike T (1998) Influences of pH and moisture content on the methane production in high-solids sludge digestion. Water Res 70: 1075-1082. https://doi.org/10.2175/106143098X123426

[20]

Czerkawski JW, Breckenridge G (1977) Design and development of a long-term rumen simulation technique (Rusitec). Br J Nutr 38: 371-384. https://doi.org/10.1079/BJN19770102

[21]

Khonkhaeng B, Cherdthong A (2020) Improving nutritive value of purple field corn residue and rice straw by culturing with white-rot fungi. J Fungi 6: 69. https://doi.org/10.3390/jof6020069

[22]

Zuo S, Niu D, Zheng M, et al. (2018) Effect of Irpex lacteus, Pleurotus ostreatus and Pleurotus cystidiosus pretreatment of corn stover on its improvement of the in vitro rumen fermentation. J Sci Food Agric 98: 4287-4295. https://doi.org/10.1002/jsfa.8952

[23]

Teoh R, Caro E, Holman DB, et al. (2019) Effects of hardwood biochar on methane production, fermentation characteristics, and the rumen microbiota using rumen simulation. Front Microbiol 10: 1534. https://doi.org/10.3389/fmicb.2019.01534

[24]

Van Soest PJ (1994) Nutritional Ecology of The Ruminant, New York: Cornell University Press. https://doi.org/10.7591/9781501732355

[25]

Royse DJ, Baars J, Tan Q (2017) Current overview of mushroom production in the world, Edible and Medicinal Mushrooms, Wiley, 5-13. https://doi.org/10.1002/9781119149446.ch2

[26]

Kholif AE, Gouda GA, Patra AK (2022) The sustainable mitigation of in vitro ruminal biogas emissions by ensiling date palm leaves and rice straw with lactic acid bacteria and Pleurotus ostreatus for cleaner livestock production. J Appl Microbiol 132: 2925-2939. https://doi.org/10.1111/jam.15432

[27]

Jenkins TC, Harfoot CG, Hazlewood GP (1993) Lipid metabolism in the rumen. J Dairy Sci 76: 382-426. https://doi.org/10.3168/jds.S0022-0302(93)77727-9

[28]

Jenkins TC, Wallace RJ, Moate PJ, et al. (2008) Board-invited review: Recent advances in biohydrogenation of unsaturated fatty acids within the rumen microbial ecosystem. J Anim Sci 86: 397-412. https://doi.org/10.2527/jas.2007-0588

[29]

Maia MRG, Chaudhary LC, Figueres L, et al. (2007) Metabolism of polyunsaturated fatty acids and their toxicity to the microflora of the rumen. Antonie Van Leeuwenhoek 91: 303-314. https://doi.org/10.1007/s10482-006-9118-2

[30]

Lu ML, Yuan GH, Li CC, et al. (2024) Effects of spent substrate of oyster mushroom (Pleurotus ostreatus) on feed utilization and liver serum indices of Hu sheep from the perspective of duodenal microorganisms. Animals 14: 3416. https://doi.org/10.3390/ani14233416

[31]

Boadi D, Benchaar C, Chiquette J, et al. (2004) Mitigation strategies to reduce enteric methane emissions from dairy cows: Update review. Can J Anim Sci 84: 319-335. https://doi.org/10.4141/A03-109

[32]

Bergman EN (1990) Energy contributions of volatile fatty acids from the gastrointestinal tract in various species. Physiol Rev 70: 567-590. https://doi.org/10.1152/physrev.1990.70.2.567

[33]

Patra AK (2013) The effect of dietary fats on methane emissions, and its other effects on digestibility, rumen fermentation and lactation performance in cattle: A meta-analysis. Livest Sci 155: 244-254. https://doi.org/10.1016/j.livsci.2013.05.023

[34]

Lourenço M, Ramos-Morales E, Wallace RJ (2010) The role of microbes in rumen lipolysis and biohydrogenation and their manipulation. Animal 4: 1008-1023. https://doi.org/10.1017/S175173111000042X

[35]

Huws SA, Kim EJ, Cameron SJS, et al. (2015) Characterization of the rumen lipidome and microbiome of steers fed a diet supplemented with flax and echium oil. Microb Biotechnol 8: 331-341. https://doi.org/10.1111/1751-7915.12164

[36]

Harfoot CG, Hazlewood GP (1997) Lipid metabolism in the rumen, The Rumen Microbial Ecosystem, Dordrecht: Springer Netherlands, 382-426. https://doi.org/10.1007/978-94-009-1453-7_9

[37]

Henderson G, Cox F, Ganesh S, et al. (2015) Rumen microbial community composition varies with diet and host, but a core microbiome is found across a wide geographical range. Sci Rep 5.

[38]

Sarnklong C, Coneja JW, Pellikaan W, et al. (2010) Utilization of rice straw and different treatments to improve its feed value for ruminants: A review. Asian Australas J Anim Sci 23: 680-692. https://doi.org/10.5713/ajas.2010.80619

[39]

Pereira AM, de Lurdes Nunes Enes Dapkevicius M, Borba AES (2022) Alternative pathways for hydrogen sink originated from the ruminal fermentation of carbohydrates: Which microorganisms are involved in lowering methane emission? Anim Microbiome 4: 5. https://doi.org/10.1186/s42523-021-00153-w

[40]

Halmemies-Beauchet-Filleau A, Rinne M, Lamminen M, et al. (2018) Review: Alternative and novel feeds for ruminants: Nutritive value, product quality and environmental aspects. Animal 12: s295-s309. https://doi.org/10.1017/S1751731118002252

[41]

Russell JB, Rychlik JL (2001) Factors that alter rumen microbial ecology. Science 292: 1119-1122. https://doi.org/10.1126/science.1058830

[42]

Mitchell KE, Lee C, Socha MT, et al. (2023) Supplementing branched-chain volatile fatty acids in dual-flow cultures varying in dietary forage and corn oil concentrations. III: Protein metabolism and incorporation into bacterial protein. J Dairy Sci 106: 7566-7577. https://doi.org/10.3168/jds.2022-23193

[43]

Liu YR, Du HS, Wu ZZ, et al. (2020) Branched-chain volatile fatty acids and folic acid accelerated the growth of Holstein dairy calves by stimulating nutrient digestion and rumen metabolism. Animal 14: 1176-1183. https://doi.org/10.1017/S1751731119002969

[44]

Bayat AR, Tapio I, Vilkki J, et al. (2018) Plant oil supplements reduce methane emissions and improve milk fatty acid composition in dairy cows fed grass silage-based diets without affecting milk yield. J Dairy Sci 101: 1136-1151. https://doi.org/10.3168/jds.2017-13545

[45]

Bayat AR, Ventto L, Kairenius P, et al. (2017) Dietary forage to concentrate ratio and sunflower oil supplement alter rumen fermentation, ruminal methane emissions, and nutrient utilization in lactating cows. Transl Anim Sci 1: 277-286. https://doi.org/10.2527/tas2017.0032

[46]

Fu R, Han L, Li Q, et al. (2025) Studies on the concerted interaction of microbes in the gastrointestinal tract of ruminants on lignocellulose and its degradation mechanism. Front Microbiol 16: 1554271. https://doi.org/10.3389/fmicb.2025.1554271

[47]

Janssen PH (2010) Influence of hydrogen on rumen methane formation and fermentation balances through microbial growth kinetics and fermentation thermodynamics. Anim Feed Sci Technol 160: 1-22. https://doi.org/10.1016/j.anifeedsci.2010.07.002

[48]

Ungerfeld EM (2020) Metabolic hydrogen flows in rumen fermentation: Principles and possibilities of interventions. Front Microbiol 11: 589. https://doi.org/10.3389/fmicb.2020.00589

[49]

Mahesh MS, Madhu M (2013) Biological treatment of crop residues for ruminant feeding: A review. Afr J Biotechnol 12: 4221-4231. https://doi.org/10.5897/AJB2012.2940

[50]

Mahesh MS (2012) Fungal bioremediation of wheat straw to improve the nutritive value and its effect on methane production in ruminants [MVSc]. National Dairy Research institute (Deemed University), Karnal, Haryana, India.

[51]

Palangi V, Kaya A, Kaya A, et al. (2022) Ecofriendly usability of mushroom cultivation substrate as a ruminant feed: Anaerobic digestion using gas production techniques. Animals 12: 1583. https://doi.org/10.3390/ani12121583

[52]

Huang X, Zhou L, You X, et al. (2023) Production performance and rumen bacterial community structure of Hu sheep fed fermented spent mushroom substrate from Pleurotus eryngii. Sci Rep 13: 1-13. https://doi.org/10.1038/s41598-023-35828-8

[53]

Wu H, Li Y, Meng Q, et al. (2021) Effect of high sulfur diet on rumen fermentation, microflora, and epithelial barrier function in steers. Animals 11: 2545. https://doi.org/10.3390/ani11092545

PDF (492KB)

18

Accesses

0

Citation

Detail

Sections
Recommended

/

〈 〉