The potential of frozen rumen fluid for ruminant feed evaluation using in vitro gas production technique

The aim of this study was to determine the potential of frozen rumen fluid (RF) for ruminant feed evaluation using in vitro gas production (IVGP) technique. This study used a factorial randomized block design with a 3 × 3 × 3 pattern, consisting three feeding regimes varying in forage to concentrate proportion, namely 20%:80% (feed A), 40%:60% (feed B), and 60%:40% (feed C), respectively; three storage time (2, 11, and 19 days); and RF from 3 breeds of indigenous cattle, namely Bali, crossbred Ongole and Madura cattle as groups. The results showed that the effects of feeding regimes, inoculum source, or storage time on total gas production, potential of gas production, rate of gas production, dry matter and organic matter degradability. Based on the results, it can be concluded that frozen RF may be used as inoculum source for feed evaluation using the IVGP technique and hence it supports the previous findings reported in the literature. Considering that the effects of different feeds, inoculum sources, and storage times gave the specific values, it is advisable that to use RF from slaughtered animals should be taken from the same breed to allow accurate comparison for feed evaluation using IVGP technique.

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In vitro gas production and its prediction on metabolize energy of complete feed using rumen fluid of three Indigenous cattle as inoculum taken from abattoir

Jurnal Ilmu-Ilmu Peternakan ◽

10.21776/ub.jiip.2021.031.02.10 ◽

2021 ◽

Vol 31 (2) ◽

pp. 168-174

Author(s):

Rizka Muizzu Aprilia ◽

Shavira Kusherawaty ◽

Hendrawan Soetanto

Keyword(s):

Rumen Fluid ◽

Block Design ◽

Gravimetric Method ◽

Insoluble Fraction ◽

Gas Production ◽

Metabolic Energy ◽

Indigenous Cattle ◽

Rumen Microbes ◽

In Vitro Gas Production

In vitro gas production (IVGP) technique has been routinely used to evaluate the nutritional value of ruminant feed. The pre-requisite of using rumen fluid from fistulae animals is now facing a raising pressure from the animal welfare organization to ban this practice. Alternatively, rumen fluid (RF) from abattoir has been suggested to be used as source of inoculum by a number of scientists. The objective of this study was to evaluate the potential of rumen fluid taken from cattle that were slaughtered at Surabaya abattoir as inoculum for IVGP measurement and its prediction on metabolic energy (ME) of a complete feed. Fresh rumen fluid of three indigenous cattle, namely Ongole Cross Breed (OCB), Madura (MC) and Bali Cattle (BC) was transferred into a vacuum thermo flask and immediately transported to the animal nutrition laboratory of Brawijaya University for further processes to measure IVGP of complete feed in a completely randomized block design with two replicates each. Gas production was monitored at time intervals and terminated after 48 hours incubation in the oxygen-free glass syringes at 38°C. Feed degradability was calculated by gravimetric method at 48 hours, while prediction of ME content was based on 24 h GP and estimated using the equation described by Menke and Steingass (1979).The results demonstrated that inoculum from OCB resulted in the superiority in all parameters, namely total GP (ml/500 mg DM), potential of GP (ml/500 mg DM), rate constant of gas production for insoluble fraction (ml/hour), and estimated ME content (MJ/DM) compared with MC and BC. Based on these findings it can be concluded that the fermentation process of complete feed in the rumen of OCB is more efficient than MC and BC which reflects to the more efficient interaction among rumen microbes to ferment the feed ingredients.

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The Use of Lactic Acid Bacteria as Ruminant Probiotic Candidates Based on In Vitro Rumen Fermentation Characteristics

Buletin Peternakan ◽

10.21059/buletinpeternak.v42i1.23317 ◽

2018 ◽

Vol 42 (1) ◽

Cited By ~ 1

Author(s):

Roni Ridwan ◽

Win Ariga Bungsu ◽

Wulansih Dwi Astuti ◽

Rohmatussolihat Rohmatussolihat ◽

Nurul Fitri Sari ◽

...

Keyword(s):

Rumen Fluid ◽

Block Design ◽

Rumen Fermentation ◽

Gas Production ◽

Serum Bottle ◽

16S Rdna Sequence ◽

Uncultured Bacteria ◽

Randomized Block Design ◽

In Vitro Rumen Fermentation

This research was conducted to select, to identify LAB isolates and to investigate the effects of the LAB as probiotics candidate in the rumen fermentation. Nine isolates exhibited the potency as candidate probiotics for cattle. The experiment was arranged in randomized block design with ten treatments and three different times of in vitro as a block. The substrate consisted of 70% forage and 30% concentrate proportion. The substrate was incubated at 39oC using serum bottle of 100 ml capacity for fermentation. Approximately 0.75 g of substrates was put inside the serum bottle glass and filled with 73 ml of buffered rumen fluid and 2 ml of LAB inoculant. Gas production was measured every 2, 4, 6, 8, 10, 12, 24, 48 and 72 h of the incubation period. Gas production kinetic was estimated by the Ørskov’s equation. The LAB with the highest gas production, as probiotics candidate, were identified using partial 16S rDNA sequence. The results of this research indicated that nine LAB produced high gas production in the range of 193-198 ml compare to that of control (173 ml). The addition of LAB in rumen fermentation resulted in digestibility 65-75%, organic matter digestibility 51-73%, and 6.67-6.68 pH. Based on the molecular identification, 8 isolates are Lactobacillus plantarum and 1 of uncultured bacteria. The LAB strain 32 L. plantarum showed the best for a ruminant probiotic candidate based on the in vitro rumen fermentation characteristic.

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Fermentabilitas ruminal secara in vitro suplementasi tepung daun gamal, kelor, randu dan sengon dalam konsentrat hijau

Jurnal Ilmu-Ilmu Peternakan ◽

10.21776/ub.jiip.2018.028.03.04 ◽

2018 ◽

Vol 28 (3) ◽

pp. 213

Author(s):

Eko Marhaeniyanto ◽

Sri Susanti

Keyword(s):

Crude Protein ◽

Block Design ◽

Gliricidia Sepium ◽

Gas Production ◽

Paraserianthes Falcataria ◽

Leaf Meal ◽

Ceiba Pentandra ◽

Randomized Block Design

The aim of this research was to study the in vitro ruminal fermentability of supplementation of Gliricidia sepium, Moringa oleifera, Lamk (MOL), Ceiba pentandra and Paraserianthes falcataria leaf meal in concentrate feed to be tested on sheep. This research with the experimental methods was arranged in Randomized Block Design (RBD) with 6 treatments and 3 groups. The treatment feed tested consisted of concentrate without leaves and concentrate with leaf meal supplementation. Crude protein content (CP) concentrates were prepared 16%, 18% and 20%. Supplementation uses a mixture of Gliricidia sepium, MOL, Ceiba pentandra and Paraserianthes falcataria leaf meal in concentrate feed (1: 1: 1: 1) as much as 10%, 20% and 30%. The measured variables were degradation of dry matter (DDM) and degradation of organic matter (DOM), gas production rate, microbial biomass and NH3 concentration. The use of mixed leaf meal in concentrate feed resulted in in vitro DDM as well as concentrate feed without leaf meal supplementation. Supplementation of leaf meal in concentrate feed as much as 30% with CP 20% resulted in a decrease in the value of fermentability. The supplementation of leaf meal in concentrate feed as much as 20% in concentrate feed with CP 18% produced the best fermentability value. Suggested for in-vivo trials on sheep using 18% protein concentrate feed by utilizing Gliricidia sepium, MOL, Ceiba pentandra and Paraserianthes falcataria leaf meal in concentrate feed as cheap protein sources.

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183 Effect of fermentation temperature on in vitro digestibility of a finishing diet

Journal of Animal Science ◽

10.1093/jas/skz053.140 ◽

2019 ◽

Vol 97 (Supplement_1) ◽

pp. 62-63

Author(s):

Miranda K Stotz ◽

Sebastian E Mejia-Turcios ◽

Andrea M Osorio ◽

Nadira Espinoza ◽

Philip M Urso ◽

...

Keyword(s):

Rumen Fluid ◽

Block Design ◽

Random Effect ◽

Gas Production ◽

In Vitro Digestibility ◽

Ph Change ◽

Fermentation Rate ◽

N Concentration ◽

Effect Of Treatment

Abstract Heat stress (HS) has been indicated to increase ruminal temperature, increase digestibility, and reduce ruminal pH of steers fed high concentrate diets. However, it is unclear if this effect is the result of greater fermentation rate, slower passage rate, or a combination. The effect of ruminal incubation temperature on substrate digestibility and rate of fermentation were evaluated. Four cannulated British-crossbreed steers (BW = 520 kg ± 30 kg) consuming an 87% (DM) concentrate diet were utilized as rumen fluid donors in a randomized complete block design with two incubation treatments (CON=39°C and HS=42°C). Within block, duplicate 125 mL serum bottles containing 200 mg of dietary substrate were inoculated with a 2:1 buffer:ruminal fluid mixture and incubated for 24 h to measure total gas production, fermentation rate, fermentation lag, pH, and ammonia-N concentration. In vitro organic matter digestibility (IVOMD) was measured separately in 100 mL centrifuge tubes. Data were analyzed using the MIXED procedure of SAS using the fixed effect of treatment and random effect of rumen fluid donor (block). There was no treatment effect on total gas production (P = 0.92) or fermentation rate (P = 0.11); however, HS began fermenting substrate sooner than CON (P < 0.005). There was a significant effect of treatment on IVOMD where HS was greater compared to CON (79.3 vs. 70.4%; P = 0.05). Final pH and relative pH change were not different (P ≥ 0.25) likely due to buffering capacity of the in vitro technique Ammonia-N concentration was greater for HS than CON (7.92 vs. 5.33 mM; P < 0.05) and may indicate a temperature effect on ruminal nitrogen availability.In conclusion, it does not appear that incubating at a greater temperature affected the fermentation rate but likely induced a change in fermentation kinetics, which may have contributed to the greater overall IVOMD

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Influence of Different Vegetable Oils on In Vitro Ruminal Fermentability and Nutrient Digestibility in Ettawah Crossbred Goat

ANIMAL PRODUCTION ◽

10.20884/1.jap.2019.21.1.689 ◽

2020 ◽

Vol 21 (1) ◽

pp. 22

Author(s):

Anis Muktiani ◽

Nurul Arifah ◽

Widiyanto Widiyanto

Keyword(s):

Vegetable Oil ◽

Palm Oil ◽

Seed Oil ◽

Corn Oil ◽

Rumen Fluid ◽

Block Design ◽

Nutrient Digestibility ◽

Dry Matter Digestibility ◽

Randomized Block Design

This research was aimed to determine the effect of supplementation of vegetable oil (corn oil, palm oil and kapok seed oil) on in vitro ruminal fermentability and nutrient digestibility. Experiment design used was Randomized Block Design with four treatments and four replication based on rumen fluid sampling time. The four treatments namely R0 = feed without oil supplementation; R1 = R0 + 5% corn oil; R2 = R0 + 5% palm oil; and R3 = R0 + 5% kapok seed oil. The result showed that supplementation of 5% vegetable oil did not affect (P>0,05) the rumen pH, A/P ratio, and efficiency of energy conversion. The total VFA, acetate, propionate, butyrate, methane and NH3 was higher (P<0,05) in supplementation of 5% vegetable oil than he control. The population of protozoa, microbial protein, in vitro dry matter digestibility (IVDMD), organic matter digestibility (IVOMD), and crude fiber digestibility (IVCFD) was lower (P<0,05) in supplementation of 5% compared to the control. Population of protozoa decreased until 58,76% for R1; 66,89% for R2; and 43,33% for R3. It can be concluded that supplementation of 5% vegetable oil decreased the population of protozoa, increased the production of VFA and NH3. Supplementation of 5% kapok seed oil resulting the highest of total VFA, acetate, propionate, butyrate and NH3 among other treatments.

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Effect of fermentation on mixed rumen contents and jackfruit peel using Aspergillus oryzae on in vitro gas production and digestibility

E3S Web of Conferences ◽

10.1051/e3sconf/202233500048 ◽

2022 ◽

Vol 335 ◽

pp. 00048

Author(s):

Mashudi Mashudi ◽

Wahyuni Nurmawati

Keyword(s):

Organic Matter ◽

Analysis Of Variance ◽

Aspergillus Oryzae ◽

Dry Matter ◽

Block Design ◽

Gas Production ◽

Dry Matter Digestibility ◽

Randomized Block Design ◽

In Vitro Gas Production

The objective of this research was to investigate the effect of Aspergillus oryzae on fermentation of mixture of rumen contents and jackfruit peel on in vitro gas production and digestibility. The method used in this study was an experiment using a randomized block design (RBD) of 4 treatments and 3 replications. The treatments including of T0 = 50% rumen contents + 50% jackfruit peel, T1 = 40 % rumen contents + 60% jackfruit peel + 0,4% Aspergillus oryzae, T2 = 30% rumen contents + 70% jackfruit peel + 0,4% Aspergillus oryzae, T3 = 20% rumen contents + 80% jackfruit peel + 0,4% Aspergillus oryzae. Variables observed were gas production, dry matter digestibility (DMD), and organic matter digestibility (OMD). Data were analyzed by using Analysis of Variance (ANOVA) from Randomized Block Design, if there were significant effect between the treatments then tested with least significant different (LSD). The result showed that fermentation of mixture of rumen contents and jackfruit peel have highly significant effect (P<0.01) on gas production, and significant effect (P<0.05) on DMD and OMD. Gas production, DMD and OMD of fermented mixture of rumen contents and jackfruit peel are higher than control without fermentation. It is concluded that the higher jackfruit peels the higher gas production, DMD and OMD.

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Effects of exogenous α-amylases, glucoamylases and proteases on ruminal in vitro dry matter and starch digestibility, gas production and volatile fatty acids of mature dent corn grain

Translational Animal Science ◽

10.1093/tas/txaa222 ◽

2020 ◽

Author(s):

Felipe X Amaro ◽

Donghyeon Kim ◽

Mariele C N Agarussi ◽

Vanessa P Silva ◽

Tatiane Fernandes ◽

...

Keyword(s):

Dry Matter ◽

Rumen Fluid ◽

Block Design ◽

Gas Production ◽

Starch Digestibility ◽

In Vivo Studies ◽

Random Factor ◽

Dent Corn ◽

Corn Grain

Abstract Two separate experiments were carried out to evaluate the effects of incremental doses of 10 exogenous endo-acting α-amylase and exo-acting glucoamylase; 1LAT (bacterial α-amylase), 2AK, 3AC, 4Cs4, 5Trga, 6Afuga, 7Fvga, and 10Tg (fungal α-amylases, glucoamylases and α-glucosidase), 8Star and 9Syn (fungal amylase-mixtures; Exp. 1) and 3 exogenous proteases; 11P14L, 12P7L, and 13P30L (bacterial proteases; Exp. 2) on in vitro dry matter digestibility (IVDMD) and in vitro starch digestibility (IVSD) of mature dent corn grain using a batch culture system. Incremental doses of the exogenous enzymes (0, 0.25, 0.50, 0.75 and 1.00 mg/g of dried substrate) were applied directly to the substrate (0.5 g of ground corn, 4 mm) in sextuplicate (Exp. 1) or quadruplicate (Exp. 2) within F57 filter bags, which were incubated at 39°C in buffered rumen fluid for 7 h. Rumen fluid was collected 2-3 h after the morning feeding from 3 lactating dairy cows and pooled. Cows were consuming a mid-lactation total mixed ration (TMR; 1.60 Mcal/kg DM and 15.4%; net energy of lactation and crude protein, respectively). Three independent runs were carried out for each experiment. Data were analyzed as a randomized complete block design using run as the blocking factor. Dose was used as a fixed factor while run was considered a random factor. Linear, quadratic, and cubic orthogonal contrasts were also tested. In Exp. 1, enzymes 2AK, 3AC and 10Tg did not increase (P > 0.10) IVDMD and IVSD, whereas 0.25 mg of enzymes 1LAT, 5Trga and 8Star increased (P < 0.01) IVDMD by 23, 47 and 62% and IVSD by 35, 41 and 58%, respectively, compared with the control. Enzymes 4Cs4, 6Afuga, 7Fvga and 9Syn linearly increased IVDMD and IVSD (P < 0.01). Greatest increases in IVDMD (82.9%) and IVSD (85.9%) resulted with 1 mg of 6Afuga compared to control. In Exp. 2, the lowest dose of exogenous proteases 11P14L and 12P7L increased (P < 0.01) IVDMD by 98 and 87% and IVSD by 57 and 64%, respectively, whereas the highest dose of 13P30L increased (P = 0.02) IVDMD by 44.8% and IVSD by 30%, relative to the control. In conclusion, IVSD and IVDMD were increased by one α-amylase, certain glucoamylases and all proteases tested, with the glucoamylase 6Afuga in Exp. 1 and the neutral protease12P7L in Exp. 2, increasing IVDMD and IVSD to the greater extents. Future in vivo studies are required to validate these findings before these enzyme additives can be recommended for improving the digestibility of mature dent corn grain.

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Degradability, in vitro fermentation parameters and kinetic degradation of diets with increasing levels of forage and chitosan

Translational Animal Science ◽

10.1093/tas/txab086 ◽

2021 ◽

Author(s):

Amanna Gonzaga Jacaúna ◽

Rafael Henrique de Tonissi e Buschinelli de Goes ◽

Leonardo de Oliveira Seno ◽

Luis Carlos Vinhas Ítavo ◽

Jefferson Rodrigues Gandra ◽

...

Keyword(s):

Degradation Kinetics ◽

Block Design ◽

Ammonia Concentration ◽

Gas Production ◽

Ruminal Fermentation ◽

In Vitro Fermentation ◽

Randomized Block Design ◽

Fermentation Parameters ◽

Potential Use

Abstract Chitosan is the second most important natural biopolymer in the world, extracted from crustaceans, shrimps, and crabs; and can modulate rumen fermentation. Our hypothesis is that the addition of chitosan alters the fermentation patterns of different diets for ruminants. This study aimed to evaluate the effects of different levels of chitosan and forage on in vitro dry degradation kinetics and fermentation in a gas production system. The chitosan levels (0, 1625, 3500 or 7500 mg/kg of DM) were arranged in a completely randomized block design, and for in vitro ruminal fermentation assay we used a split splot arrangement. Into the incubator, all chitosan levels were distributed in the four jars, and the forage levels varying on 100, 65, 50, 35 and 20 on DM basis. Chitosan and roughage levels interaction effect (P≤0.05) on IVDMD; IVOMD. IVDCP and IVDNDF. Chitosan negatively affected IVDMD in all roughage levels evaluated. The pH and ammonia concentration present effect only for roughage levels and incubation hours. The chitosan didn’t change (P=0.3631) the total short-chain fatty acid concentration (overall mean = 21.19 mmol/L) and the C2:C3 ratio (overall mean = 5.85). The IVDCP showed the same decreasing quadratic behavior (P<0.0001). The increasing chitosan addition increases (P<0.0001) the gas production and decreases the (P<0.0001) the lag time (parameter C) of diets with greater concentrate participation, characterizing greater efficiency in the degradability of the diet, confirming its potential use in diets for ruminants. Chitosan changes in vitro dry degradation kinetics and fermentation at the minimum dose of 1722 mg/kg DM for all diets. The roughage level influenced the in vitro nutrients degradability and cumulative gas production.

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Chemical composition and in vitro gas production of cassava peel silage using different preservatives

IOP Conference Series Earth and Environmental Science ◽

10.1088/1755-1315/888/1/012052 ◽

2021 ◽

Vol 888 (1) ◽

pp. 012052

Author(s):

A D Kartika ◽

Kusmartono ◽

Mashudi ◽

PH Ndaru

Keyword(s):

Chemical Composition ◽

Rice Bran ◽

Block Design ◽

Gas Production ◽

Randomized Block Design ◽

Randomized Design ◽

In Vitro Gas Production ◽

Completely Randomized Design ◽

Cassava Peel

Abstract This study aimed to evaluate the effects of using different types and levels of preservatives on chemical composition and in vitro gas production (IVGP) of cassava peel silage. The following treatments were applied: T0: cassava peel without preservative; T1: T0 + molasses 2%; T2: T0 + molasses 4%; T3: T0 + rice bran 2%; T4: T0 + rice bran 4%; T5: T0 + cassava meal 2% and T6:T0 + cassava meal 4% with 3 replications arranged in a completely randomized design (CRD) for the chemical composition variable and randomized block design (RBD) for IVGP variable. The results showed that treatments significantly affected (P<0.01) DM, OM, and EE contents, significantly affected (P<0,05) IVGP but did not significantly affect CP, CF contents, potential, and rate of gas production (P>0.05). Treatment T5 seems to have higher values of IVGP and ME (159.1±7.19 ml/500 mgDM; 11.1±0.36MJ/kg DM, respectively) compared to the other treatments that might have related to a higher OM available for in vitro fermentation process. It can be concluded that the use of cassava meal at 2% level can be considered as the most ideal preservative to use to produce a high-quality cassava peel silage.

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Weed Interference in the Establishment of Urochloa ruziziensis

Planta Daninha ◽

10.1590/s0100-83582019370100077 ◽

2019 ◽

Vol 37 ◽

Author(s):

A.A. LOURENÇO ◽

R.V. MOTA ◽

J.L. SANCHES ◽

R.F. MARQUES ◽

S.R. MARCHI

Keyword(s):

Greenhouse Gas ◽

Block Design ◽

Gas Production ◽

Co2 Production ◽

Morphological Parameters ◽

Weed Interference ◽

Randomized Block Design ◽

Dead Material ◽

Carbon Dioxide Co2

ABSTRACT: This research aimed to study weed interference relationship on morphogenesis, yield, and greenhouse gas production potential of Urochloa ruziziensis under pasture renovation conditions. The experimental design was a randomized block design with four replications. Treatments consisted of seven coexistence periods: 0, 15, 30, 45, 60, 75, and 90 days after emergence (DAE). The following morphological parameters were analyzed: number of tillers, number of leaves, photosynthetically active leaf blade biomass, fresh stem biomass, dead material biomass, and total dry biomass. In vitro analyses of methane (CH4) and carbon dioxide (CO2) production were also carried out. The results were submitted to analysis of variance by the F-test, and the test of means was carried out by the Scott-Knott test at 5%. Biomass, CH4, and CO2 production of U. ruziziensis were modified with only 15 days of weed coexistence, with an effect even higher from 45 DAE. Therefore, weeds interfere with all morphological parameters, yield, and greenhouse gas production in the pasture establishment with the forage grass U. ruziziensis.

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