scholarly journals Rhodaneses Enzyme Addition Could Reduce Cyanide Concentration and Enhance Fiber Digestibility via In Vitro Fermentation Study

Fermentation ◽  
2021 ◽  
Vol 7 (4) ◽  
pp. 207
Author(s):  
Chanadol Supapong ◽  
Anusorn Cherdthong
Keyword(s):  
Volatile Fatty Acids ◽  
Potassium Cyanide ◽  
Insoluble Fraction ◽  
Ammonia Nitrogen ◽  
Gas Production ◽  
Nutrient Digestibility ◽  
Control Group ◽  
In Vitro Digestibility ◽  
Ruminal Fermentation

The use of cyanide-containing feed (HCN) is restricted because it causes prussic acid poisoning in animals. The objective of this study was to see how adding rhodanese enzyme to an HCN-containing diet affected gas dynamics, in vitro ruminal fermentation, HCN concentration reduction, and nutrient digestibility. A 3×4 factorial arrangement in a completely randomized design was used for the experiment. Factor A was the three levels of potassium cyanide (KCN) at 300, 450, and 600 ppm. Factor B was the four doses of rhodanese enzyme at 0, 0.65, 1, and 1.35 mg/104 ppm KCN, respectively. At 96 h of incubation, gas production from an insoluble fraction (b), potential extent (omit gas) (a + b), and cumulative gas were similar between KCN additions of 300 to 450 ppm (p > 0.05), whereas increasing KCN to 600 ppm significantly decreased those kinetics of gas (p < 0.05). Supplementation of rhodanese enzymes at 1.0 to 1.35 mg/104 ppm KCN enhanced cumulative gas when compared to the control group (p < 0.05). Increasing the dose of rhodanese up to 1.0 mg/104 ppm KCN significantly increased the rate of ruminal HCN degradation efficiency (DE) by 70% (p < 0.05). However, no further between the two factors was detected on ruminal fermentation and in vitro digestibility (p > 0.05). The concentration of ammonia-nitrogen (NH3-N) increased with increasing doses of KCN (p < 0.05), but remained unchanged with varying levels of rhodanese enzymes (p > 0.05). The in vitro dry matter digestibility (IVDMD) was suppressed when increasing doses of KCH were administered at 600 ppm, whereas supplementation of rhodanese enzymes at 1.0–1.35 mg/104 ppm KCN enhanced IVDMD (p < 0.05). Increasing doses of KCN affected reduced total volatile fatty acids (TVFA) concentration, which was lowest when 600 ppm was added (p < 0.05). Nevertheless, the concentration of TVFAs increased when rhodanese enzymes were included by 1.0–1.35 mg/104 ppm KCN (p < 0.05). Based on this study, it could be concluded that supplementation of rhodaneses enzyme at 1.0–1.35 mg/104 ppm KCN could enhance cumulative gas, digestibility, and TVAF, as well as lowering ruminal HCN concentration.

scholarly journals Combining Crude Glycerin with Chitosan Can Manipulate In Vitro Ruminal Efficiency and Inhibit Methane Synthesis

Animals ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 37
Author(s):  
Anuthida Seankamsorn ◽  
Anusorn Cherdthong ◽  
Metha Wanapat
Keyword(s):  
Volatile Fatty Acids ◽  
Gas Production ◽  
Nutrient Digestibility ◽  
Crude Glycerin ◽  
Control Group ◽  
In Vitro Digestibility ◽  
Ruminal Fermentation ◽  
Ch4 Production ◽  
In Vitro Gas Production

It was hypothesized that the combination of glycerin and chitosan improves ruminal fermentation efficiency via an enhanced propionate (C3) and reduces in vitro CH4 production. This was explored through in vitro gas production with substrates containing crude glycerin, which replaced cassava chips in the studied ration. The experimental design was organized following a 3 × 3 factorial in completely randomized design and the arrangement of treatments were different levels of crude glycerin supplementations 0, 10.5, and 21% of total mixed ration (TMR) and chitosan levels were added at 0, 1, and 2% dry matter (DM) of substrate. Then, 0.5 g of TMR substrates were added into 40 mL bottles, together with respective doses of chitosan and then incubated at 39 °C. The dietary treatments were performed in three replicates within the incubation, and incubations were repeated on three separate days (runs). No interactions were found between crude glycerin and chitosan doses in terms of theoretical maximum of asymptotic gas production (b), rate of gas production (c), the discrete lag time prior to gas production (L), or the cumulative gas production at 96 h of incubation (p > 0.05). Cumulative gas production at 96 h of incubation was similar among the doses of crude glycerin and levels of chitosan, which ranged from 64.27 to 69.66 mL/g DM basis of substrate (p > 0.05). The concentration of ruminal NH3-N after 2 and 4 h of incubation ranged from 14.61 to 17.10 mg/dL and did not change with the addition of crude glycerin with chitosan (p > 0.05). The concentration of CH4 after 2 h of incubation did not change among treatments (p > 0.05), whereas after 4 h of incubation, CH4 synthesis was significantly reduced by enhancing doses of crude glycerin and chitosan (p < 0.05). The combination of 21% of crude glycerin in TMR with 2% chitosan depressed CH4 production as much as 53.67% when compared to the non-supplemented group. No significant crude glycerin and chitosan interaction effect was detected for in vitro digestibility of nutrients after incubation for 12 and 24 h using the in vitro gas production technique (p > 0.05). In addition, no significant changes (p > 0.05) were observed in total volatile fatty acids, acetate (C2) or butyrate content among treatments and between the main effects of crude glycerin with chitosan. At 4 h of incubation, ruminal C3 content and the C2 to C3 ratio changed significantly when crude glycerin and chitosan was added (p < 0.05). The 21% crude glycerin incorporate into TMR, in combination with 2% additional chitosan, increased C3 content by 26.41%, whereas the ratio of C2 to C3 was reduced by 31% when compared to the control group. Propionate concentration increased by 11.75% when increasing levels of chitosan at 2% of substrate, whereas the C2 to C3 ratio decreased by 13.99% compared to the 0% chitosan group. The inclusion of crude glycerin at 21% in TMR diets with chitosan supplementation at 2% enhanced ruminal propionate concentration and reduced methane production without causing any detrimental effect on the gas kinetics or nutrient digestibility.

Effect of sulfur concentrations in fermented total mixed rations containing fresh cassava root on rumen fermentation

2020 ◽  
Vol 60 (11) ◽  
pp. 1429 ◽  
Author(s):  
Chanadol Supapong ◽  
Anusorn Cherdthong
Keyword(s):  
Dry Matter ◽  
Nitrogen Concentration ◽  
Insoluble Fraction ◽  
Gas Production ◽  
Nutrient Digestibility ◽  
Hydrocyanic Acid ◽  
Ruminal Fermentation ◽  
Cassava Root ◽  
Dietary Treatments

Context Feeding of fresh cassava root in ruminants is limited because it contains a high level of hydrocyanic acid (HCN), which is responsible for poisoning. Aims The objective of the present study was to evaluate the effect of sulfur levels supplementation in the fermented total mixed ration (FTMR) containing fresh cassava root as an energy source on the gas kinetics, ruminal fermentation, reduction of HCN concentration and nutrient digestibility in the in vitro gas production. Methods The experimental design was a 3 × 4 factorial in a completely randomised design. Dietary treatments contained factor A, which was three levels of sulfur supplementation at 0, 1 and 2% in FTMR, and factor B was ensiling time at 0, 7, 14 and 21 days respectively. Key results Concentration of HCN in FTMR was significantly reduced (P &lt; 0.05) by 73.7% when sulfur was supplemented in FTMR at 2%. The levels of HCN in FTMR were 2.89, 0.61, 0.61 and 0.49 ppm, for ensiling time of 0, 7, 14 and 21 days, respectively (P &lt; 0.01). HCN was reduced when ensiling started at 7 days. Gas production from soluble fractions (a) ranged from –1.2 to –2.4 and was not significant (P &gt; 0.05). Furthermore, gas production from the insoluble fraction (b) ranged from 48.8 to 53.9, and gas production rate constants for the insoluble fraction (c) ranged from 0.1 to 0.2. The potential extent of gas production (a + b) was also unchanged when the concentration of sulfur increased (P &gt; 0.05). In addition, there were no interactions between sulfur levels and ensiling times on all parameters (P &gt; 0.05). In contrast, cumulative gas production (at 96 h of incubation) was significantly different when sulfur increased at 2% (P &lt; 0.05), while ensiling times did not affect cumulative gas production. Ruminal pH was affected by FTMR and decreased with an ensiling time of 21 days, ranging from 6.0 to 6.1 after ensiling. Ensiling time did not affect ruminal ammonia-nitrogen concentration (P &gt; 0.05) among dietary treatments which ranged from 21.2 to 24.0 mg%. FTMR ensiled for 21 days had the highest in in vitro dry matter digestibility, an in vitro neutral detergent fibre and in vitro acid detergent fibre digestibility which were 61.0–62.5, 35.1–43.1 and 22.3–25.9% dry matter (DM) respectively. Regarding the concentration of total volatile fatty acid (VFA), acetic acid, propionic acid and butyric acid, ranges from 94.7 to 113.6 mmol/L, 59.3 to 67.4, 20.2 to 25.9 and 11.3 to 13.8 mol/100 mol, respectively, were observed and did not differ among treatments (P &gt; 0.05). The concentration of total VFA relative to the sulfur level and ensiling time had no effect on ruminal VFA concentrations. However, exceedingly high percentages of sulfur (2% of the DM) in the diet tend to increase total VFA concentration. Conclusions Using of 2% sulfur supplementation in TMR containing fresh cassava root fermented could improve the kinetics of gas and nutrient digestibility while maintaining ruminal fermentation parameters and the rate of HCN disappearance. Implications These findings should be examined in further in vivo experiments in order to increase animal performance.

scholarly journals Effects of garlic supplementation on in vitro nutrient digestibility, rumen fermentation, and gas production

2021 ◽  
Vol 51 (2) ◽  
pp. 271-279
Author(s):  
M.R. Kekana ◽  
D. Luseba ◽  
M.C. Muyu
Keyword(s):  
Fatty Acids ◽  
Crude Protein ◽  
Dry Matter ◽  
Volatile Fatty Acids ◽  
Ammonia Nitrogen ◽  
Rumen Fermentation ◽  
Gas Production ◽  
Nutrient Digestibility ◽  
Garlic Powder

Garlic contains secondary metabolites with antimicrobial properties that can alter nutrient digestibility and rumen fermentation, similar to other antimicrobial products. The objectives of the study were to evaluate the effects of garlic powder and garlic juice on in vitro nutrient digestibility, rumen fermentation, and gas production. The treatments consisted of control with no additives, garlic powder, and garlic juice at 0.5 ml and 1 ml. The digestibility of dry matter, crude protein and neutral detergent fibre were determined after 48 hours incubation. Rumen ammonia nitrogen and volatile fatty acids were determined at 12 hours and 24 hours incubation. The cumulative gas production was recorded periodically over 48 hours. The in vitro dry matter disappearance decreased with 1 ml of garlic juice compared with control. The crude protein degradability in garlic powder and garlic juice was lower than in control. Volatile fatty acids increased in all treatments. Individual volatile fatty acids were significantly different, especially propionate, whereas the acetate to propionate ratio was reduced by garlic juice, and ammonia nitrogen was reduced by garlic powder and 0.5 ml of garlic juice. The cumulative gas production increased significantly with both levels of garlic juice. The addition of garlic juice at 0.5 mL/100 ml could enhance the production of propionate, and reduce the acetate to propionate ratio, implying that the supply of hydrogen for methanogens was limited.

Furanocoumarins in tedera do not affect ruminal fermentation in continuous culture

2015 ◽  
Vol 55 (4) ◽  
pp. 544 ◽  
Author(s):  
M. H. Ghaffari ◽  
Z. Durmic ◽  
D. Real ◽  
P. Vercoe ◽  
G. Smith ◽  
...  
Keyword(s):  
Volatile Fatty Acids ◽  
Protein Digestibility ◽  
Nutrient Digestibility ◽  
In Vitro Digestibility ◽  
Microbial Fermentation ◽  
Ruminal Fermentation ◽  
Fermentation Liquid ◽  
Acid Detergent Fibre ◽  
Sampling Times

The objective of this study was to investigate the effects of the forage shrub tedera (Bituminaria bituminosa) on nutrient digestibility, rumen microbial fermentation and furanocoumarins degradation in the rusitec. The variables were measured in fermentation liquid on Days 13 and 17 and were compared with a control (lucerne hay). Overall, tedera had greater (P < 0.05) neutral detergent fibre and acid detergent fibre digestibility than lucerne hay on both days of the experiment, but on Day 17 it had lower (P < 0.01) dry matter and crude protein digestibility than lucerne hay. There were no significant differences in concentration of NH3-N and pH between treatments, but NH3-N concentrations in both treatments were lower (P < 0.05) on Day 17 than on Day 13. The concentration of total volatile fatty acids in vessels were not affected by treatments, but the concentration of acetate was lower and acetate-to-propionate ratio higher (P < 0.05) in tedera than lucerne hay on Day 13 of the experiment. Furanocoumarins were detected in the tedera treatment only. Their concentration in the fermentation liquid increased immediately after the addition of the plant material to the fermenter, reaching highest concentrations after 2 h. These concentrations gradually declined over the next two sampling times, but 6 h after the ‘feeding’, they were still detectable in the fermentation liquid. It was concluded that (i) tedera had in vitro digestibility and fermentability variables comparable to lucerne; (ii) furanocoumarins were degraded in the fermentation fluid, and (iii) furanocoumarins from tedera did not seem to impede microbial fermentation. Tedera may provide an alternative feed source to hay and grain for filling the summer–autumn feed gap without negatively affecting nutrient digestibility and rumen microbial fermentation.

scholarly journals Residue of propolis extract in bovine diets with increasing levels of protein on rumen fermentation

2020 ◽  
Vol 55 ◽  
Author(s):  
Roberto Junior Teixeira Nascimento ◽  
Rafael Monteiro Araújo Teixeira ◽  
Thierry Ribeiro Tomich ◽  
Luiz Gustavo Ribeiro Pereira ◽  
Tânia Dayana do Carmo ◽  
...  
Keyword(s):  
Volatile Fatty Acids ◽  
Ammonia Nitrogen ◽  
Rumen Fermentation ◽  
Gas Production ◽  
Ruminal Fermentation ◽  
Propolis Extract ◽  
In Vitro Gas Production ◽  
Dry Matter Degradation ◽  
Kinetics Of

Abstract: The objective of this work was to evaluate the effect of the residue from the extraction of propolis, added to bovine diets with increasing levels of protein, on ruminal fermentation in vitro. For this, the in vitro gas production technique was used. Incubation was carried out with inocula from three fistulated cows, in three periods. In each period, a cow received a daily dose of 100 g propolis residue. Four diets were evaluated: corn silage (control); and 25, 50, and 75% concentrate based on soybean meal. The following were determined: kinetics of rumen fermentation; dry matter degradation; production of gases, volatile fatty acids (acetate, propionate, and butyrate), methane, and ammonia nitrogen; and pH. The inclusion of 14.4, 15.1, and 9.5% propolis residue, respectively, to 25, 50, and 75% concentrate increased the production of gases from the degradation of fibrous carbohydrates, when compared with the control. The propolis residue reduces methane production and the acetate:propionate ratio at all tested concentrate inclusion levels.

Efficacy of exogenous xylanases for improving in vitro fermentation of forages

2014 ◽  
Vol 153 (3) ◽  
pp. 538-553 ◽  
Author(s):  
Z. X. HE ◽  
L. Y. YANG ◽  
W. Z. YANG ◽  
K. A. BEAUCHEMIN ◽  
S. X. TANG ◽  
...  
Keyword(s):  
Volatile Fatty Acids ◽  
Xylanase Activity ◽  
Ammonia Nitrogen ◽  
Positive Control ◽  
Linear Increase ◽  
Gas Production ◽  
Pennisetum Purpureum ◽  
Ruminal Fermentation ◽  
Positive Effects

SUMMARYBatch cultures of mixed rumen micro-organisms were used to evaluate varying enzyme products with high xylanase activity (EPX), four of which were recombinant single xylanase activity developmental enzyme products (EPX1–EPX4, products of xylanase genes derived from Trichoderma harzianum, Trichoderma reesei, Orpinomyces and Aspergillus oryzae, respectively), for their potential to improve in vitro ruminal fermentation of three forages [maize (Zea mays) stover (MS), rice (Oryza sativa) straw (RS) and Guimu No. 1 grass (Pennisetum americanum×Pennisetum purpureum, GM)]. The enzyme product EPX5, derived from Trichoderma longibrachiatum, was used as a positive control that could improve in vitro fermentation of forages. Enzymes were supplied at dose rates of 0 (control), 20 (low), 50 (medium) and 80 (high) enzymic units of xylanase/g of dry matter (DM). There were no interactions between EPX and dose for the fermentation characteristics evaluated. Increasing EPX dose linearly increased gas production (GP) kinetic characters [i.e. asymptotic GP (VF), half time when GP is half of the theoretical maximum GP (t0·5), and initial fractional rate of degradation (FRD0)] and methane (CH4) production from RS and GM at 24 h, and increased degradability of DM at 24 h for MS and RS. A linear increase in degradability of neutral detergent fibre (NDF) of the three forages at 24 h was observed with increasing dose of EPX, but at 48 h only NDF degradability of RS was increased. There were differences in the effects of EPX on degradability of DM and NDF from RS at 24 h, with EPX4 having the highest and EPX1 having the lowest. In addition, increasing EPX dose linearly increased acetate proportion at 24 h and total volatile fatty acids (TVFA) at 48 h in MS. Increasing EPX dose linearly increased TVFA at 24 h, and ammonia-nitrogen (NH3-N) concentration at 48 h in RS. For GM, linear or quadratic effects of dose on acetate and butyrate concentration were observed at 24 and 48 h. The present study indicates that applying EPX to low-quality forages has the potential to improve rumen degradability and utilization. Furthermore, EPX from different sources differed in their effects when applied at the same dose rate, with the responses being forage-specific. For RS, the EPX derived from A. oryzae showed the greatest positive effects on forage degradation; whereas for MS and GM, the source of micro-organism where EPX gene was derived did not affect the degradation, with little difference among the EPX evaluated.

scholarly journals Digestibility and rumen fermentation of a high forage diet pre-treated with a mixture of cellulase and xylanase enzymes

2021 ◽  
Vol 51 (3) ◽  
pp. 399-406
Author(s):  
K. Selzer ◽  
A. Hassen ◽  
A.M. Akanmu ◽  
A.Z.M. Salem
Keyword(s):  
Volatile Fatty Acids ◽  
Rumen Fluid ◽  
Latin Square ◽  
Poor Quality ◽  
Gas Production ◽  
Nutrient Digestibility ◽  
Ruminal Fermentation ◽  
Forage Digestibility

Forages play an important role in ruminant animal production worldwide. Unlocking the nutritional potential of poor-quality tropical forages with fibrolytic enzymes would improve forage digestibility and utilization. Using in vitro and in vivo methods this study investigated the effect of pre-treating Smutsfinger hay for 24 hours with a mixture of fibrolytic enzyme (100% cellulase; 75% cellulase: 25% xylanase; 50% cellulase: 50% xylanase; 25% cellulase: 75% xylanase; 100% xylanase and a control with no enzyme) on ruminal fermentation and digestibility of nutrients by sheep. For in vitro fermentation, dry matter, neutral detergent fibre (NDF) degradability and volatile fatty acids (VFA) were determined with standard procedures. The same treatments were used for an in vivo digestibility trial using Merino sheep in a 6 x 6 Latin square design. Feed intake and total tract digestibility were recorded. Rumen fluid samples were collected daily, preserved, and analysed for VFA. The addition of 100% cellulase enzyme to Smutsfinger hay in vitro increased (P <0.05) NDF degradability and gas production compared with the control and inclusion of 100% xylanase enzyme. Both 100% cellulase and xylanase enzymes significantly reduced in vitro end time fermentation pH. A 50:50 mixture of cellulase and xylanase plus enzyme in vivo, increased acetate, total VFA concentration, and higher NDF and ADF digestibility of the test feed compared with the control. Inclusion of a 50-75% mixture of cellulase and 50-25% xylanase enzymes treatment led to higher gas production and butyrate concentration, decreased ruminal pH and improved nutrient digestibility.

scholarly journals Effects of garlic supplementation on in vitro nutrient digestibility, rumen fermentation and gas production.

2021 ◽  
Author(s):  
M. R. Kekana ◽  
D. Luseba ◽  
M. C. Muyu
Keyword(s):  
Fatty Acids ◽  
Crude Protein ◽  
Dry Matter ◽  
Volatile Fatty Acids ◽  
Ammonia Nitrogen ◽  
Rumen Fermentation ◽  
Gas Production ◽  
Nutrient Digestibility ◽  
Garlic Powder

Abstract Garlic has secondary metabolites that possess antimicrobial properties that can alter nutrient digestibility and rumen fermentation similar to known antimicrobial products. The objectives of the study were to evaluate the effects of garlic powder and garlic juice on in vitro nutrient digestibility, rumen fermentation and gas production. The treatments were control no additives, garlic powder and garlic juice of 0.5 ml and 1 ml. The digestibility of dry matter, crude protein and neutral detergent fibre were determined after 48h incubation. Rumen ammonia nitrogen and volatile fatty acids were determined at 12h and 24h incubation. The cumulative gas production was recorded periodically over 48h. The in vitro dry matter disappearance decreased with 1 ml of garlic juice compared to control. The crude protein degradability in garlic powder and garlic juice was lower than in control. The volatile fatty acids increased in all the treatments. Individual volatile fatty acids were significantly different, especially propionate, while the acetate:propionate ratio was reduced by garlic juice, and ammonia nitrogen was reduced by garlic powder and 0.5 ml of garlic juice. The cumulative gas production increased significantly with both levels of garlic juice. The addition of garlic juice at 0.5 mL/100 mL could enhance the production of propionate, and reduce the acetate:propionate ratio implying that the supply of hydrogen for methanogens is limited.

scholarly journals Pomegranate seed oil rich in conjugated linolenic acids reduces in vitro methane production

1970 ◽  
Vol 46 (3) ◽  
pp. 325-335
Author(s):  
E. Maleki ◽  
G.Y. Meng ◽  
M. Faseleh Jahromi ◽  
R. Jorfi ◽  
A. Khoddami ◽  
...  
Keyword(s):  
Methane Production ◽  
Volatile Fatty Acids ◽  
Seed Oil ◽  
Control Diet ◽  
Nutrient Utilization ◽  
Gas Production ◽  
Metabolizable Energy ◽  
Ruminal Fermentation ◽  
Ch4 Production

The objective of this study was to determine the effect of pomegranate (Punica granatum L.) seed oil (PSO) on gas and methane (CH4) production, ruminal fermentation and microbial populations under in vitro conditions. Three treatments consisting of a control diet containing 10 mg tallow (CON); the control diet with 5 mg PSO + 5 mg tallow (MPSO) and the control diet containing 10 mg PSO (HPSO) were compared. Ten mg of the experimental fat/oil samples were inserted into a gas-tight 100 mL plastic syringe containing 30 mL of an incubation inoculum and 250 mg of a basic substrate of a hay/concentrate (1/1, w/w) mixture. In vitro gas production was recorded over 0, 2, 4, 6, 8, 10, 12 and 24 h of incubation. After 24 hours, incubation was stopped, and methane production, pH, volatile fatty acids (VFAs) and microbial counts were measured in the inoculant. Gas production at 4, 6, 8, 10, 12 and 24 h incubation, metabolizable energy and in vitro organic matter disappearance increased linearly and quadratically as level of PSO increased. Furthermore, the 10 mg PSO (HPSO) decreased CH4 production by 21.0% compared with the control (CON) group. There were no significant differences in total and individual VFA concentrations between different levels of PSO, except for butyric acid. After 24 h of incubation, methanogenesis decreased in the HPSO compared with the MPSO and CON treatments. In addition, total bacteria and protozoa counts increased with rising PSO levels, while population methanogenesis declined significantly. These results suggested that PSO could reduce methane emissions, which might be beneficial to nutrient utilization and growth in ruminants.

scholarly journals Using glycerin with chitosan extracted from shrimp residue to enhance rumen fermentation and feed use in native Thai bulls

2021 ◽  
pp. 1158-1164
Author(s):  
Anuthida Seankamsorn ◽  
Anusorn Cherdthong ◽  
Sarong So ◽  
Metha Wanapat
Keyword(s):  
Volatile Fatty Acids ◽  
Latin Square ◽  
Ammonia Nitrogen ◽  
Rumen Fermentation ◽  
Nutrient Digestibility ◽  
Crude Glycerin ◽  
Ruminal Fermentation ◽  
Design Factor ◽  
Ch4 Production ◽  
High Level

Background and Aim: Crude glycerin is changed to propionate in the rumen, while chitosan can be used as a feed supplement to increase propionic acid concentration and decrease methane (CH4) production. We hypothesized that supplementation with a combination of a high level of crude glycerin with chitosan could have a beneficial effect on ruminal fermentation and mitigate CH4 production. This study aimed to explore the combined effects of crude glycerin and chitosan supplementation on nutrient digestibility, rumen fermentation, and CH4 calculation in native Thai bulls. Materials and Methods: Four 2-year-old native Thai bulls, weighing 150±20 kg, were kept in a 2×2 factorial arrangement in a 4×4 Latin square design. Factor A represented the incorporation of crude glycerin at 10.5% and 21% of the dry matter (DM) of a total mixed ration (TMR), and factor B represented the supplementation of chitosan at 1% and 2% DM of a TMR. Results: Increasing levels of crude glycerin at 21% decreased DM intake by 0.62 kg/day compared with 10.5% crude glycerin (p<0.05), whereas nutrient digestibility did not change (p>0.05). The incorporated crude glycerin and supplemented chitosan levels did not affect the pH, temperature, concentrations of ammonia-nitrogen, microbial population, and blood urea nitrogen (p>0.05). Supplemented chitosan and incorporated crude glycerin did not show any interaction effects on the molar portions and total volatile fatty acids (VFAs), except estimated CH4. Increasing the incorporated crude glycerin levels increased propionate and decreased the ratio of acetate to propionate ratio, whereas levels of butyrate, acetate, and total VFAs were unchanged. The combination of crude glycerin at 21% in the TMR with chitosan at 2% reduced CH4 estimation by 5.08% compared with the other feed treatment. Conclusion: Increasing incorporated crude glycerin levels in a TMR significantly elevated the propionate concentration, whereas combining 21% crude glycerin in the TMR diet with 2% chitosan supplementation could depress CH4 estimation more effectively than adding one of these supplements alone.

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