Effect of Solid State Fermented (SSF) Biomass on In Vitro Methanogenesis and Dry Matter Digestibility in Adult Surti Buffaloes

Effect of Solid State Fermented (SSF) Biomass on In Vitro Methanogenesis and Dry Matter Digestibility in Adult Surti Buffaloes | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Effect of Solid State Fermented (SSF) Biomass on In Vitro Methanogenesis and Dry Matter Digestibility in Adult Surti Buffaloes Dr. Gaurang Pravinbhai Mathukiya, Dr. Paresh R. Pandya, Dr. Kalpesh K. Sorathiya, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5139548/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract An in vitro rumen fermentation study was conducted to evaluate the effect of supplementation with solid state fermented biomass (SSF) in total mixed rations (TMRs) on total gas production, methanogenesis and dry matter digestibility by using the rumen liquor of adult Surti buffaloes. SSF biomass was supplemented at 0, 1, 2, 3, 4, 5, 6, 7, and 8% with TMR (65% wheat straw and 35% concentrate) for in vitro gas production trials. The results of the in vitro study revealed significantly (P < 0.05) greater IVDMD (58.43%) and lower CH 4 production (3.58 ml CH 4 /100 mg DDM) at a 3% level of SSF biomass addition in TMR. Based on the overall results of in vitro studies, 3% SSF biomass supplementation was found to be most suitable for further in vivo studies in adult Surti buffaloes. Animal Science SSF Biomass In Vitro Digestibility Methane Gas Production 1. INTRODUCTION Improvements in feed utilization, animal production, health, and food safety are the goals of rumen microbial research. These goals can be accomplished by encouraging ideal fermentation, minimizing ruminal problems, and preventing infections. It is best to think of supplements as an addition to sound feeding procedures. An animal's behavior can be influenced by a class of feed chemicals known as feed additives, which are only required at trace levels. The use of anti biowaste agents in feed has substantially decreased in the last ten years due to the detrimental effects of these agents on animal health, the residue they left in animal products, and the possibility that microbes could become resistant to them. As a result, the use of microorganisms in animal nutrition has gained popularity. The development of enzyme supplements that enhance fiber digestion and lower enteric methane emissions from large ruminants has been the main emphasis of research. The two main techniques for extracting enzymes are solid-state fermentation (SSF) and submerged fermentation (SmF). Due to its decreased energy need, decreased effluent generation, and direct application of fermented products for feeding, the bioconversion of fibrous material by SSF has attracted increasing interest [Yang et al. 2011 ] and [Surabhiet al. 2024 ]. Fermentation involving solids in the absence (or almost absence) of free water is referred to as "solid-state fermentation." For the generation of enzymes by microbial flora, solid-state fermentation has enormous promise. This method is particularly intriguing because it results in a raw, fermented product that may be employed immediately as an enzyme source [C. Senthil kumar & R. Vijay Anand et al. 2023]. Ideally, the SSF system can be used to manufacture practically every known microbial enzyme. The production of enzymes such as proteases, cellulases, xylanases, amylases, and pectinases has been the subject of extensive research [Pandey et al. 1999 ] and [Partha Sarathi Patra and Ashim Chandra Sinha et al. 2012]. Exogenous fibrinolytic enzymes have significantly increased digestibility and decreased intestinal methane emission and positive effects on increasing the efficiency of feed utilization by ruminants both in vitro [Murad et al. 2009 ] and in vivo [Arriola et al. 2011 ] in recent years as a result of being added to animal feeds. Therefore, the present study investigated the effect of SSF biomass supplementation on in vitro rumen fermentation. 2. MATERIALS AND METHODS The present study was conducted at the Animal Nutrition Research Station, College of Veterinary Science and Animal Husbandry, Kamdhenu University, Anand, Gujarat. Wheat straw, mung gotar, groundnut cake, deoiled rice bran, molasses and mineral mixture were used for preparation of the total mixed rations (TMRs). The TMR was oven dried at 70°C and finely ground in a Wiley mill using 1 mm sieve. The TMR was analyzed for proximate constituents (AOAC, 2005 ) and fiber fraction [Van Soest et al. 1991 ] SSF biomass was procured from the Department of Microbiology, Gujarat Vidhyapeeth, Sadra, Gandhinagar, Gujarat, India. The solid-state fermented (SSF) biomass of jowar hay was carried out with a culture of Aspergillus oryzae and Trichoderma spp. fungi. The experimental TMR without any supplementation of the SSF biomass was the control group and was designated as S0, while the TMR with SSF biomass supplementation at 1, 2, 3, 4, 5, 6, 7 and 8% was designated as S1, S2, S3, S4, S5, S6, S7 and S8, respectively. Rumen liquor for in vitro rumen fermentation studies was collected from two adult Surti buffaloes using stomach tubes. The buffaloes were fed individually with TMRs prepared to meet their nutrient requirements (ICAR 2013) with free access to water. The collected rumen liquor was strained through four layers of muslin cloth, which was termed the strained rumen liquor (SRL), and was mixed in prepared artificial saliva (McDougall’s) in proper proportions prior to incubation. Substrates (200 mg) with various levels of SSF biomass were incubated with artificial saliva mixed with SRL (40 ml) for 48 h in quadrupling at 39 ± 1°C in a shaker twin water bath [Menke et al. 1979 ]. After 48 h of incubation, total gas production (TGP) was recorded after subtracting the gas production from the blank. To determine in vitro methane production, gas produced in 100 ml glass syringes after a 24 h of incubation period was used. A gas sample was directly injected into gas chromatograph (GC) from each syringe, and the CH 4 concentration was determined against standard methane gas (22.54%). All the samples were analyzed using a GC instrument, fitted with an SS column (4 ft. long, 3.2 mm inside diameter) packed with Porapack N (80 to 100 mesh), and equipped with a flame ionization detector (FID). The column temperature was maintained at 50°C, and nitrogen was used as the carrier gas, with a flow rate of 30 ml/min. Calibration was completed using standards (22.54%) procured from CHEMIX Specialty Gases & Equipment., Bangalore. After completion of incubation, the content of each syringe was filtered and dried in a preweighed Gooch crucible. The IVDM was calculated by subtracting the residues remaining after incubation from the amount of substrate incubated and was expressed as a percentage. Statistical analysis The data generated during the experiment were analyzed by two-way analysis of variance (ANOVA) using the WASP 2.0 method as prescribed by [Snedecor and Cochran 1994 ]. 3. RESULTS AND DISCUSSION The proximate composition and fiber fraction data of the prepared TMRs are presented in Table 1 . The effects of solid-state fermented (SSF) biomass on IVDMD, total gas production (TGP) and methane (ml/100 g of digestible DM) are presented in Table 2 . Table 1 Chemical Composition and Fiber Fraction of the Total Mixed Ration Parameters (%, on DM basis) TMR Crude protein 11.08 Ether extract 2.33 Crude fiber 27.42 Nitrogen free extract 45.30 Total Ash 13.87 Organic matter 78.84 Neutral detergent fiber 53.13 Acid detergent fiber 35.32 Cellulose 28.12 Hemicellulose 17.81 Lignin 5.62 Calcium 1.50 Table 2 Average In Vitro Dry Matter Digestibility (IVDMD, %), Total Gas Production (TGP, Ml) and Methane (Ml/100 G Digestible DM) of Substrates Containing Different Levels of SSF Biomass Substrates IVDMD TGP Methane S0 53.12 ab ± 0.37 67.67 ± 2.96 4.01 ± 0.44 S1 48.97 a ± 0.35 35.33 ± 15.07 4.01 ± 0.77 S2 51.90 ab ± 2.78 59.00 ± 11.27 3.85 ± 0.83 S3 58.43 b ± 4.53 52.33 ± 11.79 3.58 ± 0.22 S4 50.80 ab ± 1.53 42.00 ± 14.00 4.02 ± 0.21 S5 52.83 ab ± 0.88 23.37 ± 14.66 4.27 ± 0.22 S6 48.03 a ± 1.27 58.00 ± 4.04 4.35 ± 0.36 S7 56.83 ab ± 2.90 50.33 ± 14.15 4.23 ± 0.22 S8 56.73 ab ± 5.70 64.00 ± 3.06 4.32 ± 0.49 SEM 3.03 11.24 0.48 CD(0.05) 8.99 NA NA CV% 9.88 38.76 20.14 * a, b, c superscripts in a column differ significantly (P < 0.05) The findings indicated that the in vitro dry matter digestibility percentages were 53.12, 48.97, 51.90, 58.43, 50.80, 52.83, 48.03, 56.83, and 56.73 for the S0, S1, S2, S3, S4, S5, S6, S7, and S8 groups, respectively. For the S0, S1, S2, S3, S4, S5, S6, S7, and S8 groups, the corresponding in-vitro total gas production (ml) values were as follows: 67.67, 35.33, 59.00, 52.33, 42.00, 23.37, 58.00, 50.33, and 64.00. The in vitro methane production values for the S0, S1, S2, S3, S4, S5, S6, S7, and S8 groups were 4.01, 4.01, 3.85, 3.58, 4.02, 4.27, 4.35, 4.23, and 4.32, respectively. A significantly (P < 0.05) greater percentage of patients in the 3% level of SSF biomass-supplemented group (58.43%) IVDMD than did those in the control group (53.12%). The addition of SSF biomass, which may have improved digestibility, resulted in higher fermentation rates in the present study. Additionally, the addition of enzymes enhances the growth of cellulolytic bacteria, accelerates the digestion of fiber, and facilitates the movement of microbial proteins from the rumen [Azzaz et al. 2013 ]. Similarly, significant (P < 0.05) improvements in IVDMD [Bhasker et al. 2012 ; Arati, 2013 ; Reddy et al. 2016 ; Chaudhari, 2018 ] were also reported by earlier authors. A total amount of gas produced during the in-vitro incubation study of 48 h was observed, and the data are presented in Table 2 . The values for total gas production were not significantly different. Methane is a substantial contributor to greenhouse gas emissions,; thus, mitigating its effects is also necessary for a better environment. The addition of probiotics, additives, or various nutrients to feed has an established effect on methane (CH 4 ) generation. [Mamuad et al. 2014 , Lamba et al. 2014 ]. The effect of SSF biomass supplementation on methane production was found to numerically (P > 0.05) lower than that of the control on methane production by 21% in the 3% SSF biomass supplemented group. 4. CONCLUSION It was observed that SSF biomass at the 3% level had a significantly greater IVDMD. At the 3% level of SSF, biomass supplementation numerically decreased in vitro methane production; hence, SSF biomass supplemented at the 3% level in the diet is the most suitable for further in vivo studies. Declarations The permission for animal experiments was granted by the Institutional Animal Ethics Committee (IAEC: 375/ANRS/2022). Funding The research grant was received from the project of Kamdhenu University. Data availability The authors acknowledge that the data presented in this study will be available upon reasonable request. Conflict of interest All the authors affirm that they have no conflicts of interest. Author contributions All the contributors contributed to the conception and planning of the study. Material preparation, data collection and analysis were performed by Dr. Gaurang P. Mathukiya, Dr. Paresh R. Pandya, Dr. Kalpesh K. Sorathiya and Dr. Sunil V. Rathod. The first draft of the manuscript was written by Dr. Gaurang P. Mathukiya, and all theauthors commented on previous versions of the manuscript. All the authors read and approved the final manuscript. Acknowledgment The authors are grateful to the Animal Nutrition Research Station, College of Veterinary Science and Animal Husbandry, Anand. I am thankful for providing the necessary facilities and financial support for undertaking this study. References AOAC (2005) Official method of Analysis. 18th Edition, Association of Official Analytical Chemists, Washington DC, Method 935.14 and 992.24. Arati P (2013) Effect of solid state fermentation (SSF) biomass on digestibility and nutrient utilization in goats [M.V.Sc. thesis, Anand Agricultural University]. Krishikosh. http://krishikosh.egranth.ac.in/handle/1/5810050977 Arriola KG, Kim SC Staples CR and Adesogan AT (2011) Effect of fibrolytic enzyme application to low and high concentrate diets on the performance of lactating dairy cattle. Journal of Dairy Science 94: 832-841. https://doi.org/10.3168/jds.2010-3424 Azzaz HH, Murad HA, Kholif AM, Morsy TA, Mansour AM, & El-Sayed HM (2013) Increasing nutrients bioavailability by using fibrolytic enzymes in dairy buffaloes feeding. Beauchemin KA, Colombatto D, Morgavi DP and Yang WZ (2003) Use of exogenous fibrolytic enzymes to improve feed utilization by ruminants. Journal of Animal Science 81: E37-E47. https://doi.org/10.2527/2003.8114_suppl_2E37x Bhasker TV, Nagalakshmi D & Rao DS, (2012) Exogenous fibrolytic enzyme cocktail for improvement of nutrient utilization from sorghum stover in cattle. Indian Journal of Dairy Science 65(4): 325-328. Chaudhari KI, (2018) Methane mitigation in crossbred calves by feeding legume straw based total mixed ration with SSF biomass [M. V. Sc. thesis, Anand Agricultural University]. Krishikosh. http://krishikosh.egranth.ac.in/handle/1/5810148755 Lamba JS, Wadhwa M, Bakshi MPS, (2014) In vitro methane production and in Sacco degradability of processed wheat and rice straws. Indian J. Anim. Nutr. 31: 345–350. Mamuad L, Kim SH, Jeong CD, Choi YJ, Jeon CO, Lee SS (2014) Effect of fumarate reducing bacteria on in vitro rumen fermentation, methane mitigation and microbial diversity. J. Microbiol. 52: 120–128. Maurya MS, Singh R, Pathak NN and Kamra DN, (1993) Effect of feeding live yeast (Sachharomyces cerevisiae) on nutrient digestibility in goats. In Proceedings Sixth Animal Nutrition Research Workshop Conference, Bhubaneswar, Sept. 13-16: 142. Menke KH, Raab L, Salewski A, Steingass H, Fritz D and Schneider W (1979) The estimation of the digestibility and metabolisable energy content of ruminant feeding stuffs from the gas production when they are incubated with rumen liquor. The Journal of Agricultural Science 93: 211-222. https://doi.org/10.1017/S0021859600086305 Murad HH Hanfy MA, Kholif AM, Abdel Gawad MH and Murad HA (2009) Effect of cellulases supplementation to some low quality roughages on digestion and milk production by lactating goats. Journal of Biological Chemistry and Environmental Sciences 4: 791-809. Pandey A, Selvakumar P, Soccol RC and Nigam P (1999) Solid state fermentation for the production of industrial enzyme. Current Science 149-162. https://www.jstor.org/stable/24102923 Partha Sarathi Patra and Ashim Chandra Sinha (2012) Studies on organic cultivation of groundnut (Arachis hypogaea) in Cooch Behar. Indian Journal of Agronomy 57 (4): 386-389. Reddy PR, Kumar DS, Rao ER, & Rao KA (2016) Nutritional evaluation of Total mixed rations supplemented with exogenous fibrolytic enzymes and/or live yeast culture in Buffalo Bulls. Indian Journal of Animal Nutrition 33(1): 54-58. 10.5958/2231-6744.2016.00009.8 Senthil kumar C & Vijay Anand R (2023) A Review of Energy-Efficient Secured Routing Algorithm for IoT-Enabled Smart Agricultural Systems, Journal of Biosystems Engineering, Volume 48, pages 339–354. Snedecor GW & Cochran WG (1994) Statistical methods. (8th Ed.). The IOWA State University press, Ames, Iowa, USA. Surabhi S, Aayushi S, & Kirtiraj D (2024) Assessment of Environmental Condition and Its Effect on Biological Parameters of Farm Women. National Academy Science Letters 1-4. Van Soest PJ, Robertson JB & Lewis BA (1991) Methods of dietary fiber. Neutral detergent fiber and non-starch polysaccharides in relation to animal nutrition. Journal of Dairy Science 74: 3583-3597. https://doi.org/10.3168/jds.S0022-0302(91)78551-2 Yang HE, Son YS and Beauchemin KA (2011) Effects of exogenous enzymes on ruminal fermentation and degradability of alfafa hay and rice straw. Journal of Animal Science 24 (1): 56-64. Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5139548","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":358086437,"identity":"8a25c534-4d5c-4065-839a-34f1cb044803","order_by":0,"name":"Dr. Gaurang Pravinbhai Mathukiya","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABFElEQVRIiWNgGAWjYHACNgaGAxIILj+ISCjAp4MZTYtkA0iLAUEtSHwDMAePFt3288ce85yxSOzvX/xMuqLmjt3m86sTPzwwYJDnFzuAVYvZmWR2Y54bEokzbjwzkzxz7FnythtvN0sAHWY4c3YCdi0HktmkeT5IJDbcOGAm2cB2ONnsxtkNIC0JBrdxaDn/GKJl/o3j3yQb/h1ONp5xdvMPvFpugGwBOmzD+R4zyca2w3YG/L3b8Nty47GZ5JwzEsYbb/AUWzb2HU6QuMG7zSLBQAK3X84nPpN4c6xOdt754xtvNnw7bM/ff3bzzR8VNvL80ti1wIBjgwREQSKUIYFHMQTYM/AfQGWMglEwCkbBKIABAHjjatGhPLuGAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0009-0009-2289-7537","institution":"College of Veterinary Science and Animal Husbandry, Anand","correspondingAuthor":true,"prefix":"Dr.","firstName":"Gaurang","middleName":"Pravinbhai","lastName":"Mathukiya","suffix":""},{"id":358086948,"identity":"8a24fc14-995a-4410-98ce-baadff0f6ffc","order_by":1,"name":"Dr. Paresh R. 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INTRODUCTION","content":"\u003cp\u003eImprovements in feed utilization, animal production, health, and food safety are the goals of rumen microbial research. These goals can be accomplished by encouraging ideal fermentation, minimizing ruminal problems, and preventing infections. It is best to think of supplements as an addition to sound feeding procedures. An animal's behavior can be influenced by a class of feed chemicals known as feed additives, which are only required at trace levels. The use of anti biowaste agents in feed has substantially decreased in the last ten years due to the detrimental effects of these agents on animal health, the residue they left in animal products, and the possibility that microbes could become resistant to them. As a result, the use of microorganisms in animal nutrition has gained popularity.\u003c/p\u003e \u003cp\u003eThe development of enzyme supplements that enhance fiber digestion and lower enteric methane emissions from large ruminants has been the main emphasis of research.\u003c/p\u003e \u003cp\u003eThe two main techniques for extracting enzymes are solid-state fermentation (SSF) and submerged fermentation (SmF). Due to its decreased energy need, decreased effluent generation, and direct application of fermented products for feeding, the bioconversion of fibrous material by SSF has attracted increasing interest [Yang et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2011\u003c/span\u003e] and [Surabhiet al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2024\u003c/span\u003e]. Fermentation involving solids in the absence (or almost absence) of free water is referred to as \"solid-state fermentation.\"\u003c/p\u003e \u003cp\u003eFor the generation of enzymes by microbial flora, solid-state fermentation has enormous promise. This method is particularly intriguing because it results in a raw, fermented product that may be employed immediately as an enzyme source [C. Senthil kumar \u0026amp; R. Vijay Anand et al. 2023]. Ideally, the SSF system can be used to manufacture practically every known microbial enzyme. The production of enzymes such as proteases, cellulases, xylanases, amylases, and pectinases has been the subject of extensive research [Pandey et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1999\u003c/span\u003e] and [Partha Sarathi Patra and Ashim Chandra Sinha et al. 2012]. Exogenous fibrinolytic enzymes have significantly increased digestibility and decreased intestinal methane emission and positive effects on increasing the efficiency of feed utilization by ruminants both \u003cem\u003ein vitro\u003c/em\u003e [Murad et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2009\u003c/span\u003e] and \u003cem\u003ein vivo\u003c/em\u003e [Arriola et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2011\u003c/span\u003e] in recent years as a result of being added to animal feeds. Therefore, the present study investigated the effect of SSF biomass supplementation on \u003cem\u003ein vitro\u003c/em\u003e rumen fermentation.\u003c/p\u003e"},{"header":"2. MATERIALS AND METHODS","content":"\u003cp\u003eThe present study was conducted at the Animal Nutrition Research Station, College of Veterinary Science and Animal Husbandry, Kamdhenu University, Anand, Gujarat. Wheat straw, mung gotar, groundnut cake, deoiled rice bran, molasses and mineral mixture were used for preparation of the total mixed rations (TMRs). The TMR was oven dried at 70\u0026deg;C and finely ground in a Wiley mill using 1 mm sieve. The TMR was analyzed for proximate constituents (AOAC, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2005\u003c/span\u003e) and fiber fraction [Van Soest et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e1991\u003c/span\u003e]\u003c/p\u003e \u003cp\u003eSSF biomass was procured from the Department of Microbiology, Gujarat Vidhyapeeth, Sadra, Gandhinagar, Gujarat, India. The solid-state fermented (SSF) biomass of jowar hay was carried out with a culture of Aspergillus oryzae and Trichoderma spp. fungi. The experimental TMR without any supplementation of the SSF biomass was the control group and was designated as S0, while the TMR with SSF biomass supplementation at 1, 2, 3, 4, 5, 6, 7 and 8% was designated as S1, S2, S3, S4, S5, S6, S7 and S8, respectively.\u003c/p\u003e \u003cp\u003eRumen liquor for \u003cem\u003ein vitro\u003c/em\u003e rumen fermentation studies was collected from two adult Surti buffaloes using stomach tubes. The buffaloes were fed individually with TMRs prepared to meet their nutrient requirements (ICAR 2013) with free access to water. The collected rumen liquor was strained through four layers of muslin cloth, which was termed the strained rumen liquor (SRL), and was mixed in prepared artificial saliva (McDougall\u0026rsquo;s) in proper proportions prior to incubation. Substrates (200 mg) with various levels of SSF biomass were incubated with artificial saliva mixed with SRL (40 ml) for 48 h in quadrupling at 39\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C in a shaker twin water bath [Menke et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e1979\u003c/span\u003e]. After 48 h of incubation, total gas production (TGP) was recorded after subtracting the gas production from the blank. To determine \u003cem\u003ein vitro\u003c/em\u003e methane production, gas produced in 100 ml glass syringes after a 24 h of incubation period was used. A gas sample was directly injected into gas chromatograph (GC) from each syringe, and the CH\u003csub\u003e4\u003c/sub\u003e concentration was determined against standard methane gas (22.54%). All the samples were analyzed using a GC instrument, fitted with an SS column (4 ft. long, 3.2 mm inside diameter) packed with Porapack N (80 to 100 mesh), and equipped with a flame ionization detector (FID). The column temperature was maintained at 50\u0026deg;C, and nitrogen was used as the carrier gas, with a flow rate of 30 ml/min. Calibration was completed using standards (22.54%) procured from CHEMIX Specialty Gases \u0026amp; Equipment., Bangalore. After completion of incubation, the content of each syringe was filtered and dried in a preweighed Gooch crucible. The IVDM was calculated by subtracting the residues remaining after incubation from the amount of substrate incubated and was expressed as a percentage.\u003c/p\u003e \u003cp\u003e \u003cb\u003eStatistical analysis\u003c/b\u003e \u003c/p\u003e \u003cp\u003eThe data generated during the experiment were analyzed by two-way analysis of variance (ANOVA) using the WASP 2.0 method as prescribed by [Snedecor and Cochran \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e1994\u003c/span\u003e].\u003c/p\u003e"},{"header":"3. RESULTS AND DISCUSSION","content":"\u003cp\u003eThe proximate composition and fiber fraction data of the prepared TMRs are presented in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The effects of solid-state fermented (SSF) biomass on IVDMD, total gas production (TGP) and methane (ml/100 g of digestible DM) are presented in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eChemical Composition and Fiber Fraction of the Total Mixed Ration\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"2\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParameters (%, on DM basis)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTMR\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCrude protein\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e11.08\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eEther extract\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCrude fiber\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e27.42\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eNitrogen free extract\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e45.30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal Ash\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e13.87\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eOrganic matter\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e78.84\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eNeutral detergent fiber\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e53.13\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAcid detergent fiber\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e35.32\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCellulose\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e28.12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eHemicellulose\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e17.81\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLignin\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5.62\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCalcium\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.50\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAverage \u003cem\u003eIn Vitro\u003c/em\u003e Dry Matter Digestibility (IVDMD, %), Total Gas Production (TGP, Ml) and Methane (Ml/100 G Digestible DM) of Substrates Containing Different Levels of SSF Biomass\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSubstrates\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIVDMD\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTGP\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMethane\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eS0\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e53.12\u003csup\u003eab\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e67.67\u0026thinsp;\u0026plusmn;\u0026thinsp;2.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eS1\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e48.97\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e35.33\u0026thinsp;\u0026plusmn;\u0026thinsp;15.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.77\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eS2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e51.90\u003csup\u003eab\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;2.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e59.00\u0026thinsp;\u0026plusmn;\u0026thinsp;11.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.83\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eS3\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e58.43\u003csup\u003eb\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;4.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e52.33\u0026thinsp;\u0026plusmn;\u0026thinsp;11.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eS4\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e50.80\u003csup\u003eab\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;1.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e42.00\u0026thinsp;\u0026plusmn;\u0026thinsp;14.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eS5\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e52.83\u003csup\u003eab\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;0.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e23.37\u0026thinsp;\u0026plusmn;\u0026thinsp;14.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eS6\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e48.03\u003csup\u003ea\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;1.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e58.00\u0026thinsp;\u0026plusmn;\u0026thinsp;4.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eS7\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e56.83\u003csup\u003eab\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;2.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e50.33\u0026thinsp;\u0026plusmn;\u0026thinsp;14.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eS8\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e56.73\u003csup\u003eab\u003c/sup\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;5.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e64.00\u0026thinsp;\u0026plusmn;\u0026thinsp;3.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.49\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSEM\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e11.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.48\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCD(0.05)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNA\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCV%\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e38.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20.14\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003e* a, b, c superscripts in a column differ significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05)\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe findings indicated that the \u003cem\u003ein vitro\u003c/em\u003e dry matter digestibility percentages were 53.12, 48.97, 51.90, 58.43, 50.80, 52.83, 48.03, 56.83, and 56.73 for the S0, S1, S2, S3, S4, S5, S6, S7, and S8 groups, respectively. For the S0, S1, S2, S3, S4, S5, S6, S7, and S8 groups, the corresponding \u003cem\u003ein-vitro\u003c/em\u003e total gas production (ml) values were as follows: 67.67, 35.33, 59.00, 52.33, 42.00, 23.37, 58.00, 50.33, and 64.00. The \u003cem\u003ein vitro\u003c/em\u003e methane production values for the S0, S1, S2, S3, S4, S5, S6, S7, and S8 groups were 4.01, 4.01, 3.85, 3.58, 4.02, 4.27, 4.35, 4.23, and 4.32, respectively.\u003c/p\u003e \u003cp\u003eA significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) greater percentage of patients in the 3% level of SSF biomass-supplemented group (58.43%) IVDMD than did those in the control group (53.12%). The addition of SSF biomass, which may have improved digestibility, resulted in higher fermentation rates in the present study. Additionally, the addition of enzymes enhances the growth of cellulolytic bacteria, accelerates the digestion of fiber, and facilitates the movement of microbial proteins from the rumen [Azzaz et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2013\u003c/span\u003e]. Similarly, significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) improvements in IVDMD [Bhasker et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Arati, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Reddy et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Chaudhari, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2018\u003c/span\u003e] were also reported by earlier authors.\u003c/p\u003e \u003cp\u003eA total amount of gas produced during the \u003cem\u003ein-vitro\u003c/em\u003e incubation study of 48 h was observed, and the data are presented in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The values for total gas production were not significantly different. Methane is a substantial contributor to greenhouse gas emissions,; thus, mitigating its effects is also necessary for a better environment. The addition of probiotics, additives, or various nutrients to feed has an established effect on methane (CH\u003csub\u003e4\u003c/sub\u003e) generation. [Mamuad et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e, Lamba et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2014\u003c/span\u003e]. The effect of SSF biomass supplementation on methane production was found to numerically (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05) lower than that of the control on methane production by 21% in the 3% SSF biomass supplemented group.\u003c/p\u003e"},{"header":"4. CONCLUSION","content":"\u003cp\u003eIt was observed that SSF biomass at the 3% level had a significantly greater IVDMD. At the 3% level of SSF, biomass supplementation numerically decreased \u003cem\u003ein vitro\u003c/em\u003e methane production; hence, SSF biomass supplemented at the 3% level in the diet is the most suitable for further \u003cem\u003ein vivo\u003c/em\u003e studies.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eThe permission for animal experiments was granted by the Institutional Animal Ethics Committee (IAEC: 375/ANRS/2022).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe research grant was received from the project of Kamdhenu University.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors acknowledge that the data presented in this study will be available upon reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the\u0026nbsp;authors affirm that they have no conflicts of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the contributors\u0026nbsp;contributed to the conception and planning of the study. Material preparation, data collection and analysis were performed by Dr. Gaurang P. Mathukiya, Dr. Paresh R. Pandya, Dr. Kalpesh K. Sorathiya and Dr. Sunil V. Rathod. The first draft of the manuscript was written by Dr. Gaurang P. Mathukiya, and all theauthors commented on previous versions of the manuscript. All the\u0026nbsp;authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgment\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors are grateful to the Animal Nutrition Research Station, College of Veterinary Science and Animal Husbandry, Anand. I am thankful for providing the necessary facilities and financial support for undertaking this study.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAOAC (2005) Official method of Analysis. 18th Edition, Association of Official Analytical Chemists, Washington DC, Method 935.14 and 992.24.\u003c/li\u003e\n\u003cli\u003eArati P (2013) Effect of solid state fermentation (SSF) biomass on digestibility and nutrient utilization in goats [M.V.Sc. thesis, Anand Agricultural University]. Krishikosh. http://krishikosh.egranth.ac.in/handle/1/5810050977\u003c/li\u003e\n\u003cli\u003eArriola KG, Kim SC Staples CR and Adesogan AT (2011) Effect of fibrolytic enzyme application to low and high concentrate diets on the performance of lactating dairy cattle. Journal of Dairy Science 94: 832-841. https://doi.org/10.3168/jds.2010-3424\u003c/li\u003e\n\u003cli\u003eAzzaz HH, Murad HA, Kholif AM, Morsy TA, Mansour AM, \u0026amp; El-Sayed HM (2013) Increasing nutrients bioavailability by using fibrolytic enzymes in dairy buffaloes feeding.\u003c/li\u003e\n\u003cli\u003eBeauchemin KA, Colombatto D, Morgavi DP and Yang WZ (2003) Use of exogenous fibrolytic enzymes to improve feed utilization by ruminants. Journal of Animal Science 81: E37-E47. https://doi.org/10.2527/2003.8114_suppl_2E37x\u003c/li\u003e\n\u003cli\u003eBhasker TV, Nagalakshmi D \u0026amp; Rao DS, (2012) Exogenous fibrolytic enzyme cocktail for improvement of nutrient utilization from sorghum stover in cattle. Indian Journal of Dairy Science 65(4): 325-328.\u003c/li\u003e\n\u003cli\u003eChaudhari KI, (2018) Methane mitigation in crossbred calves by feeding legume straw based total mixed ration with SSF biomass [M. V. Sc. thesis, Anand Agricultural University]. Krishikosh. http://krishikosh.egranth.ac.in/handle/1/5810148755\u003c/li\u003e\n\u003cli\u003eLamba JS, Wadhwa M, Bakshi MPS, (2014) In vitro methane production and in Sacco degradability of processed wheat and rice straws. Indian J. Anim. Nutr. 31: 345\u0026ndash;350.\u003c/li\u003e\n\u003cli\u003eMamuad L, Kim SH, Jeong CD, Choi YJ, Jeon CO, Lee SS (2014) Effect of fumarate reducing bacteria on in vitro rumen fermentation, methane mitigation and microbial diversity. J. Microbiol. 52: 120\u0026ndash;128.\u003c/li\u003e\n\u003cli\u003eMaurya MS, Singh R, Pathak NN and Kamra DN, (1993) Effect of feeding live yeast (Sachharomyces cerevisiae) on nutrient digestibility in goats. In Proceedings Sixth Animal Nutrition Research Workshop Conference, Bhubaneswar, Sept. 13-16: 142.\u003c/li\u003e\n\u003cli\u003eMenke KH, Raab L, Salewski A, Steingass H, Fritz D and Schneider W (1979) The estimation of the digestibility and metabolisable energy content of ruminant feeding stuffs from the gas production when they are incubated with rumen liquor. The Journal of Agricultural Science 93: 211-222. https://doi.org/10.1017/S0021859600086305\u003c/li\u003e\n\u003cli\u003eMurad HH Hanfy MA, Kholif AM, Abdel Gawad MH and Murad HA (2009) Effect of cellulases supplementation to some low quality roughages on digestion and milk production by lactating goats. Journal of Biological Chemistry and Environmental Sciences 4: 791-809.\u003c/li\u003e\n\u003cli\u003ePandey A, Selvakumar P, Soccol RC and Nigam P (1999) Solid state fermentation for the production of industrial enzyme. 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The IOWA State University press, Ames, Iowa, USA.\u003c/li\u003e\n\u003cli\u003eSurabhi S, Aayushi S, \u0026amp; Kirtiraj D (2024) Assessment of Environmental Condition and Its Effect on Biological Parameters of Farm Women. National Academy Science Letters 1-4.\u003c/li\u003e\n\u003cli\u003eVan Soest PJ, Robertson JB \u0026amp; Lewis BA (1991) Methods of dietary fiber. Neutral detergent fiber and non-starch polysaccharides in relation to animal nutrition. Journal of Dairy Science 74: 3583-3597. https://doi.org/10.3168/jds.S0022-0302(91)78551-2\u003c/li\u003e\n\u003cli\u003eYang HE, Son YS and Beauchemin KA (2011) Effects of exogenous enzymes on ruminal fermentation and degradability of alfafa hay and rice straw. Journal of Animal Science 24 (1): 56-64.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"Kamdhenu University","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"SSF Biomass, In Vitro, Digestibility, Methane, Gas Production","lastPublishedDoi":"10.21203/rs.3.rs-5139548/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5139548/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eAn \u003cem\u003ein vitro\u003c/em\u003e rumen fermentation study was conducted to evaluate the effect of supplementation with solid state fermented biomass (SSF) in total mixed rations (TMRs) on total gas production, methanogenesis and dry matter digestibility by using the rumen liquor of adult Surti buffaloes. SSF biomass was supplemented at 0, 1, 2, 3, 4, 5, 6, 7, and 8% with TMR (65% wheat straw and 35% concentrate) for \u003cem\u003ein vitro\u003c/em\u003e gas production trials. The results of the \u003cem\u003ein vitro\u003c/em\u003e study revealed significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) greater IVDMD (58.43%) and lower CH\u003csub\u003e4\u003c/sub\u003e production (3.58 ml CH\u003csub\u003e4\u003c/sub\u003e/100 mg DDM) at a 3% level of SSF biomass addition in TMR. Based on the overall results of \u003cem\u003ein vitro\u003c/em\u003e studies, 3% SSF biomass supplementation was found to be most suitable for further \u003cem\u003ein vivo\u003c/em\u003e studies in adult Surti buffaloes.\u003c/p\u003e","manuscriptTitle":"Effect of Solid State Fermented (SSF) Biomass on In Vitro Methanogenesis and Dry Matter Digestibility in Adult Surti Buffaloes","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-10-07 09:15:20","doi":"10.21203/rs.3.rs-5139548/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"31344930-fb4c-44a1-99c6-be7f4db08f3e","owner":[],"postedDate":"October 7th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":38114597,"name":"Animal Science"}],"tags":[],"updatedAt":"2024-10-07T09:15:20+00:00","versionOfRecord":[],"versionCreatedAt":"2024-10-07 09:15:20","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5139548","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5139548","identity":"rs-5139548","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}