In vitro rumen gas production kinetics, hydrocyanic acid concentration and fermentation characteristics of fresh cassava root and feed block sulfur concentration

2020 ◽  
Vol 60 (5) ◽  
pp. 659 ◽  
Author(s):  
Gamonmas Dagaew ◽  
Anusorn Cherdthong ◽  
Metha Wanapat ◽  
Pin Chanjula
Keyword(s):  
Nitrogen Concentration ◽  
Gas Production ◽  
Nutrient Digestibility ◽  
Hydrocyanic Acid ◽  
In Vitro Digestibility ◽  
High Concentration ◽  
Cassava Root ◽  
Fermentation Liquor ◽  
Kinetics Of

Context Feeding ruminants with fresh cassava roots (FCR) is limited because they have a high concentration of hydrocyanic acid (HCN). Thus, it was hypothesised that receiving a feed block containing high sulfur (FBS) would reduce hydrocyanic acid (HCN) in FCR and improve rumen fermentation and nutrient digestibility in animals. Aims The goal of the present work was to study the influence of the ratio of FCR to rice straw (RS) together with FBS on kinetics of gas production, HCN concentration, fermentation characteristics and nutrient digestibility, using in vitro technique. Methods The experimental design was a 4 × 2 factorial arrangement in a completely randomised design, with three replications per treatment. Factor A was the FCR to RS ratio, which was 100:0, 60:40, 40:60 or 0:100. Factor B was sulfur, which was provided as two concentrations in FBS (2% and 4% DM). Gas production was recorded after incubation, at 0, 0.5, 1, 2, 4, 6, 8, 12, 18, 24, 48, 72 and 96 h. Fermentation liquor was collected and determined for kinetics of gas production, HCN concentration, fermentation characteristics and nutrient digestibility. Key results Cassava root to RS ratio influenced the cumulative gas production after 96 h. Inclusion of sulfur in the FBS at 4% increased the cumulative gas production, when compared with inclusion at 2%. The gas production from degradable fraction (b) and the rate of gas production (c) were significantly different among the treatments with different FCR:RS ratios, whereas there was no difference between using 2% and 4% sulfur in the FBS. The HCN concentration in fermentation liquor increased with an increasing proportion of FCR. Furthermore, inclusion of sulfur in the feed block at 4% reduced HCN concentration by 42.8%, when compared with inclusion at 2% (P < 0.05). Ammonia-nitrogen concentration was significantly different among the FCR:RS-ratio treatments and was reduced when the proportion of FCR was decreased (P < 0.05). In vitro digestibility was significantly increased with an increasing proportion of FCR. Increasing the proportion of FCR with 4% of sulfur in the FBS significantly increased in vitro DM digestibility, compared with 2% sulfur. Increasing the FCR:RS ratio with 4% of sulfur in the FBS increased the proportion of propionic acid (P < 0.05). Conclusions Using a high FCR:RS ratio (100:0 or 60:40) with 4% sulfur in the FBS enhanced kinetics of gas production, propionic molar proportion, nutrient digestibility, and HCN detoxification by rumen microbes in an in vitro trial. Implications An in vivo study should be encouraged to verify the results and obtain more data.

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 < 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 < 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 > 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 > 0.05). In addition, there were no interactions between sulfur levels and ensiling times on all parameters (P > 0.05). In contrast, cumulative gas production (at 96 h of incubation) was significantly different when sulfur increased at 2% (P < 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 > 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 > 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 133 Porcine in vitro degradation and fermentation characteristics of heat-pretreated and multi-enzyme-supplemented soybean hulls

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 114-115
Author(s):  
Cienna J Boss ◽  
Jung Wook Lee ◽  
Rob Patterson ◽  
Tofuko A Woyengo
Keyword(s):  
In Vitro Digestion ◽  
Gas Production ◽  
In Vitro Digestibility ◽  
Soybean Hulls ◽  
In Vitro Fermentation ◽  
Heat Pretreatment ◽  
Fractional Rate ◽  
Enzyme Supplementation ◽  
Kinetics Of

Abstract A study was conducted to determine effects of pretreating and supplementing soybean hulls with multi-enzyme on porcine in vitro digestion and fermentation characteristics. Treatments were untreated and heat-pretreated (160 °C and 70 psi for 20 min) soybean hulls without or with multi-enzyme in a 2 × 2 factorial arrangement. The multi-enzyme supplied 2,800 U of cellulase, 1,800 U of pectinase, 400 U of mannanase, 1,000 U of xylanase, 600 U of glucanase, and 200 U of protease/kilogram of feedstuff. Feedstuffs were subjected to in vitro digestion with porcine pepsin and pancreatin, followed by in vitro fermentation for 72 h. Accumulated gas production was recorded and modeled to estimate kinetics of gas production. On DM basis, untreated and pretreated soybean hulls contained 10.4 and 10.6% CP, and 63.2 and 49.5% ADF, respectively. Pretreatment and multi-enzyme supplementation did not interact on in vitro digestibility of DM (IVDDM). Untreated and pretreated soybean hulls did not differ in IVDDM (24.8 vs. 25.7%). Multi-enzyme increased (P < 0.05) IVDDM of soybean hulls by a mean of 45.5%. Pretreatment and multi-enzyme unaffected total gas production. Pretreatment and multi-enzyme interacted (P < 0.05) on fractional rate of degradation such that the fractional rate of degradation for pretreated soybean hulls was greater (P < 0.05) than that of untreated soybean hulls when soybean hulls were supplemented with multi-enzyme (0.045 vs. 0.062 h-1), but not when soybean hulls were unsupplemented with multi-enzyme (0.053 vs. 0.059 h-1). In conclusion, multi-enzyme supplementation increased IVDDM, implying that the multi-enzyme used in the study can be used to enhance utilization of soybean hulls. Heat pretreatment increased the rate of fermentation of multi-enzyme-supplemented soybean hulls, implying that the rate of fermentation of soybean hulls in the hindgut of pigs can be enhanced by a combination of heat pretreatment and multi-enzyme supplementation.

Prediction of Digestibility and Intake of Sheep Fed Leguminosae or GRAMINAE Hays: Comparison Between the in Vitro Digestibility, Kinetics of Gas Production or Dry Matter Degradation

1994 ◽  
Vol 1994 ◽  
pp. 103-103
Author(s):  
M.T. Dentinho ◽  
K. Khazaal ◽  
J.M. Ribeiro ◽  
E.R. Ørskov
Keyword(s):  
Dry Matter ◽  
Lolium Multiflorum ◽  
Daily Intake ◽  
Gas Production ◽  
Italian Ryegrass ◽  
In Vitro Digestibility ◽  
Kinetics Of

By using separated values of kinetics of in situ dry matter (DM) degradation or in vitro gas production (Menke and Steingass, 1988) of leguminosae hays, Khazaal et al, (1993) reported high correlation with intake (r= 0.88; r= 0.79) and in vivo DM digestibility (DMD) (r= 0.94; r= 0.88). The aim of the present study was to extend the range of samples used and compare the ability of the 2 stages in vitro digestibility (Tilley and Terry, 1963), the in situ DM degradation or the gas production techniques to predict daily intake (g DM/ kgW0.75) and in vivo DM digestibility (DMD) of 19 leguminous and graminaceous hays fed to sheep.Three harvesting stages (early bloom EB, mid bloom MB or in seed IS) made from lucerne (Medicago sativa), sweet clover (Melilotus segetalis), Persian clover (Trifolium resupinatum), Rye (Secale cereale), Triticale (Triticale hexaploid), oat (Avena stativa) and a pre-bloom (PB) Italian ryegrass (Lolium multiflorum ). Each hay was fed ad libitum to 4 Merino male sheep and their intake and in vivo DMD recorded. Gas production (ml/ 200 mg DM) or in situ DM degradation (g/ 100 g DM) were determined as described by Khazaal et al, (1993) after 6, 12, 24, 48, 72 or 96 h incubation. Measured gas production or DM degradation values were fitted to the equation p=a+b(l-e-ct)(McDonald, 1981) where p is gas production or DM degradation at time t and a, b and c are constants. For nylon bag the washing loss (soluble fraction) was defined as A, the insoluble but fermentable matter was defined as B=(a+b)-A, and c is the rate of fermentation or degradation (Ørskov and Ryle, 1990).

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.

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 Pre-incubation of ruminal inocula to assess in vitro gas production and digestibility

2020 ◽  
Vol 50 (8) ◽  
Author(s):  
Juan Manuel Cantet ◽  
Darío Colombatto ◽  
Marisa Wawrzkiewicz ◽  
Gustavo Jaurena
Keyword(s):  
Gas Production ◽  
In Vitro Digestibility ◽  
Fermentation Products ◽  
In Vitro Gas Production ◽  
Production Techniques ◽  
Ruminal Degradation ◽  
Fermentative Activity ◽  
Fermentation Parameters ◽  
Kinetics Of

ABSTRACT: In vitro gas production techniques represent a valuable tool to describe the kinetics of ruminal degradation of food. However, the ruminal liquor used as a microbial inoculum has been a great source of variation and error. A standardization of this factor should contribute to assure the independence of food fermentation parameters from those of the inocula. In this research it was hypothesized that a controlled pre-incubation treatment of ruminal liquor could contribute to stabilize and homogenize the undigested residues of blanks and as a consequence, of the production of residual cumulative gas production (CGP). A pre-incubation (i.e. previous real incubation) of rumen inocula was developed with a simple substrate similar to the diet offered to donors at 1% w/v for 0, 1, 2 and 4 h (Control, Prei-1, Prei-2 and Prei-4 treatments respectively). Once the pre-incubation hours were completed, they were incubated with contrasting substrates and without substrate (i.e. blanks) in order to evaluate the CGP, in vitro digestibility of the DM and fermentation products. Although, the fermentative activity of the pre-incubated inoculums worked satisfactorily in the in vitro system, contrary to what was speculated, residues of the pre-incubation increased the variability and heterogeneity of variances among blanks. Consequently, it was concluded that the pre-incubations did not work to generate more homogeneous and less variable ruminal liquor for the in vitro gas production system.

Effect of adding different levels of probiotic on in vitro gas production

2009 ◽  
Vol 2009 ◽  
pp. 187-187
Author(s):  
M Besharati ◽  
A Taghizadeh ◽  
A Ansari
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Nutritive Value ◽  
Cellulose Degradation ◽  
Gas Production ◽  
Nutrient Digestibility ◽  
Production Performance ◽  
Attractive Alternative ◽  
Kinetics Of ◽  
Different Levels

Probiotics present an attractive alternative to the use of chemical and hormonal promoters in the livestock growth production industry. Preparations that contain micro-organisms have been safely used for many years and are generally accepted by both the farmer and the final consumer. Saccharomyces cerevisiae supplementation in ruminant diets can increase DMI, production performance, cellulose degradation, and nutrient digestibility (Callaway and Martin, 1997). The gas measuring technique has been widely used for the evaluation of nutritive value of feeds. Gas measurement provides useful data on digestion kinetics of both soluble and insoluble fractions of feedstuffs (Getachew et al., 1998). In the gas method, kinetics of fermentation can be studied on a single sample and therefore a relatively small amount of sample is required or a larger number of samples can be evaluated at time. The purpose of this study was to study effect of adding different levels of Saccharomyces cerevisiae on in vitro gas production from a dried grape by-product.

scholarly journals Anti-nutritional potential of protodioscin and kinetics of degradation in Urochloa grasses

2016 ◽  
Vol 37 (4) ◽  
pp. 2247 ◽  
Author(s):  
Eduardo Souza Leal ◽  
Luís Carlos Vinhas Ítavo ◽  
Cacilda Borges do Valle ◽  
Camila Celeste Brandão Ferreira Ítavo ◽  
Alexandre Menezes Dias ◽  
...  
Keyword(s):  
Crude Protein ◽  
Dry Matter ◽  
Nutritional Value ◽  
Gas Production ◽  
In Vitro Digestibility ◽  
Leaves Of Grass ◽  
Exotic Grasses ◽  
Kinetics Of Degradation ◽  
Kinetics Of

This study aimed to evaluate the nutritional value of Urochloa exotic grasses to quantify aspects of anti-quality and to identify their correlation with quality grasses. We evaluated the grasses U. humidicola cv. Comum, U. humidicola cv. BRS Tupi, U. decumbens cv. Basilisk, D70 U. decumbens and U. ruziziensis ecotype R124, and we grouped them into summer, autumn, winter and spring. We determined the chemical composition, in vitro digestibility, levels of protodioscin and cumulative gas production from the leaves of grass materials in nature. Basilisk and D70 showed higher content protodioscin in all seasons, with the highest values (31.4 and 27.4 g kg-1, respectively) in spring. D70 had a crude protein content of 140.0 g kg-1 in summer and a better in vitro digestibility of dry matter (888.7 g kg-1). R124 had a higher cumulative gas production in the spring (16.44 mL gas 100 mg DM-1). U. humidicola (Comum and BRS Tupi) presented lower protodioscin concentrations (1.22 and 1.07 g kg DM-1, respectively), and U. decumbens (Basilisk and D70) presented higher concentrations (27.25 and 24.55 g kg DM-1, respectively). The presence of protodioscin interfered with in vitro digestibility results and cumulative gas production in vitro.

scholarly journals 126 Porcine in vitro digestion and fermentation characteristics of heat and multi-enzyme pretreated whole stillage

2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 50-50
Author(s):  
Kevin S Jerez Bogota ◽  
Tofuko A Woyengo
Keyword(s):  
In Vitro Digestion ◽  
Gas Production ◽  
In Vitro Digestibility ◽  
In Vitro Fermentation ◽  
Heat Pretreatment ◽  
Percentage Points ◽  
Whole Stillage ◽  
Kinetics Of ◽  
Different Sources

Abstract Effects of heat pretreatment (HT) and multi-enzyme predigestion (MP) of whole stillage on porcine in vitro digestibility of DM (IVDDM) and fermentation characteristics of WS were investigated. Four WS samples were obtained from 4 different sources. Half amount of WS from each source was pretreated at 70psi and 160°C for 20 min. Untreated and pretreated WS samples from each source were divided into 4 sub-samples (4 sub-samples of untreated WS per source and 4 sub-samples of pretreated WS per source) to give 32 sub-samples. Four treatments were applied to 32 sub-samples WS (1 untreated or 1 pretreated sub-sample per treatment per sample source). The treatments were WS undigested or pre-digested with 1 of 3 multi-enzymes (MTE1, MTE2, and MTE3). The MTE1 contained xylanase, β-glucanase, cellulase, mannanase, protease, and amylase; MTE2 contained xylanase, α-galactosidase, and celullase; and MTE3 contained xylanase, cellulase, β-glucanase, and mannanase. The 32 sub-samples were subjected to porcine in vitro digestion in 3 cycles of 2 batches (16 sub-samples/batch). Subsequently, residues were subjected to porcine in vitro fermentation for 72 hours, during which accumulated gas production was recorded and modeled to estimate kinetics of gas production. The IVDDM of untreated WS was 73.4%. HT improved (P&lt; 0.05) of WS IVDMM by 8.2 percentage points. MP improved IVDDM of untreated WS and heat-pretreated WS by a means 9.1 and 6.8 percentage points, respectively. However, the magnitude of improvement in IVDDM of pretreated WS due to predigestion was lower (P&lt; 0.05) for MTE3 than that for MTE2 (4.8 vs. 9.0 percentage points), but similar to that for MTE1 (6.7 percentage points). Similar interactions were observed for total gas production. In conclusion, the digestibility of WS was improved by the HT and MP. Combination of HT and MTE2 predigestion was the most effective in improving digestibility of WS.

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