• Title/Summary/Keyword: In vitro Rumen Degradability

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Dietary lysophospholipids supplementation inhibited the activity of lipolytic bacteria in forage with high oil diet: an in vitro study

  • Kim, Hanbeen;Kim, Byeongwoo;Cho, Seongkeun;Kwon, Inhyuk;Seo, Jakyeom
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.10
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    • pp.1590-1598
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    • 2020
  • Objective: The objective of this study was to evaluate the effects of lysophospholipids (LPL) supplementation on rumen fermentation, degradability, and microbial diversity in forage with high oil diet in an in vitro system. Methods: Four experimental treatments were used: i) annual ryegrass (CON), ii) 93% annual ryegrass +7% corn oil on a dry matter (DM) basis (OiL), iii) OiL with a low level (0.08% of dietary DM) of LPL (LLPL), and iv) OiL with a high level (0.16% of dietary DM) of LPL (HLPL). An in vitro fermentation experiment was performed using strained rumen fluid for 48 h incubations. In vitro DM degradability (IVDMD), in vitro neutral detergent fiber degradability, pH, ammonia nitrogen (NH3-N), volatile fatty acid (VFA), and microbial diversity were estimated. Results: There was no significant change in IVDMD, pH, NH3-N, and total VFA production among treatments. The LPL supplementation significantly increased the proportion of butyrate and valerate (Linear effect [Lin], p = 0.004 and <0.001, respectively). The LPL supplementation tended to increase the total bacteria in a linear manner (p = 0.089). There were significant decreases in the relative proportions of cellulolytic (Fibrobacter succinogenes and Ruminococcus albus) and lipolytic (Anaerovibrio lipolytica and Butyrivibrio proteoclasticus) bacteria with increasing levels of LPL supplementation (Lin, p = 0.028, 0.006, 0.003, and 0.003, respectively). Conclusion: The LPL supplementation had antimicrobial effects on several cellulolytic and lipolytic bacteria, with no significant difference in nutrient degradability (DM and neutral detergent fiber) and general bacterial counts, suggesting that LPL supplementation might increase the enzymatic activity of rumen bacteria. Therefore, LPL supplementation may be more effective as an antimicrobial agent rather than as an emulsifier in the rumen.

The Effect of Yerba Mate (Ilex Paraguariensis) Supplementation on Nutrient Degradability in Dairy Cows: An In sacco and In vitro Study

  • Hartemink, Ellen;Giorgio, Daniela;Kaur, Ravneet;Di Trana, Adriana;Celi, Pietro
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.11
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    • pp.1606-1613
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    • 2015
  • This study was carried out to investigate the effects of Yerba Mate (YM) supplementation on nutrients' degradation, in vitro dry matter disappearance, gas production and rumen ammonia concentration. Three rumen-fistulated Holstein Friesian cows were used for the in situ incubations and provided rumen liquor for in vitro incubations. The inclusion of YM in a control diet (pasture+pellets) affected some in sacco degradation parameters. YM supplementation decreased the effective degradability and degradation rate of pasture crude protein (CP), and it seems to slow down the degradation of pasture neutral detergent fiber. A significant increase of degradation of pasture acid detergent fiber (ADF) was detected after YM inclusion in the control diet. YM supplementation reduced in vitro gas production of pasture and ammonia concentration of pellets. The addition of YM in ruminant diet could decrease ammonia production and increase protein availability for productive purposes. The moderate presence of tannins in YM could have affected the degradation kinetics of pasture CP and ADF and the ammonia production of pellets.

Effect of flaking on the digestibility of corn in ruminants

  • Kang, Hamin;Lee, Mingyung;Jeon, Seoyoung;Lee, Sang Moon;Lee, Ju Hwan;Seo, Seongwon
    • Journal of Animal Science and Technology
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    • v.63 no.5
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    • pp.1018-1033
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    • 2021
  • In this study, we aimed to assess the effect of flaking on the nutrient digestibility of corn grain in ruminants. In this regard, in vitro rumen fermentation, in situ rumen degradability, and in vivo metabolic experiments were performed. The automated gas production technique was used for the in vitro fermentation experiments. Six types of corn flakes with various degrees of gelatinization (32%, 41%, 48%, 66%, 86%, and 89%) were ground and incubated in rumen fluid to measure rumen fermentation characteristics and digestion rate. The in situ degradability of ground corn, whole corn, and corn flakes with 62% and 66% gelatinization was measured by incubation in the rumen of two cannulated Holstein cows. In vivo metabolic experiments were performed using 12 crossbred goats (29.8 ± 4.37 kg) using a 3 × 3 Latin square design. The dietary treatments consisted of ground corn and flaked corn with 48% or 62% gelatinization. In vitro experiments showed that as the degree of gelatinization increased, the digestion rate increased linearly, while the discrete lag time decreased linearly (p < 0.05). The effective rumen dry matter degradability, determined by in situ fermentation, was 37%p lower in corn flakes than ground corn, assuming a passage rate of 6%/h (p < 0.01), and there was no difference between the two flakes. In the in vivo experiment, there was no difference in dry matter intake, average daily gain, feed efficiency, and nitrogen utilization among the treatment groups (p > 0.05); however, the crude fat digestibility was lower for corn flakes than for ground corn (p < 0.05). To summarize, the rate of fermentation of corn flakes increased as the degree of gelatinization increased. However, non-ground corn flakes had lower rumen digestibility and did not improve in vivo apparent nutrient digestibility, compared with ground corn. In contrast to the assumption that flaked corn provides more energy to ruminant animals than ground corn, we conclude that the digestibility and energy value of corn flakes are lower than those of ground corn if mastication does not sufficiently reduce the particle size of corn flakes.

Evaluation of Some Aquatic Plants from Bangladesh through Mineral Composition, In Vitro Gas Production and In Situ Degradation Measurements

  • Khan, M.J.;Steingass, H.;Drochner, W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.4
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    • pp.537-542
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    • 2002
  • A study was conducted to evaluate the nutritive potential value of different aquatic plants: duckweed (Lemna trisulaca), duckweed (Lemna perpusila), azolla (Azolla pinnata) and water-hyacinth (Eichhornia crassipes) from Bangladesh. A wide variability in protein, mineral composition, gas production, microbial protein synthesis, rumen degradable nitrogen and in situ dry matter and crude protein degradability were recorded among species. Crude protein content ranged from 139 to 330 g/kg dry matter (DM). All species were relatively high in Ca, P, Na, content and very rich in K, Fe, Mg, Mn, Cu and Zn concentration. The rate of gas production was highest in azolla and lowest in water-hyacinth. A similar trend was observed with in situ DM degradability. Crude protein degradability was highest in duckweed. Microbial protein formation at 24 h incubation ranged from 38.6-47.2 mg and in vitro rumen degradable nitrogen between 31.5 and 48.4%. Based on the present findings it is concluded that aquatic species have potential as supplementary diet to livestock.

Predicting In Sacco Rumen Degradation Kinetics of Raw and Dry Roasted Faba Beans (Vicia faba) and Lupin Seeds (Lupinus albus) by Laboratory Techniques

  • Yu, P.;Egan, A.R.;Leury, B.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.10
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    • pp.1377-1387
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    • 2000
  • Two laboratory techniques: (1) an in vitro method with two procedures for measuring protein degradabilities and (2) an in vitro method with three procedures for measuring protein solubility, were investigated to determine which laboratory techniques could most accurately predict the quantity of rumen protein degradation kinetics of legume seeds after dry roasting under various conditions, in terms of (1) rumen protein disappearance ($D_j$, where j=0, 2, 4, 8, 12, 24 and 48 h incubation), (2) rumen protein effective degradability (EDCP), (3) the parameters describing rumen degradation characteristics (the soluble fraction: S, the potentially degradable fraction: D, undegradable fraction: U, lag time: T0 and the degradation rate: Kd) and (4) rumen bypass protein (BCP), which were determined by the method accepted internationally at present, in sacco nylon bag technique using the standardized Dutch method. Feeds evaluated were the raw and dry roasted whole faba (Vicia faba) beans (WFB) and whole lupin (Lupinus albus) seeds (WLS), each was dry roasted under various conditions (at 110, 130 or $150^{\circ}C$ for 15, 30 or 45 min). In vitro protein degradability ($D_1$_Auf and $D_{24}$_Auf) were determined using the modified Aufr re method by enzymatic hydrolysis for 1 h and 24 h using a protease extracted from Streptomyces griseus in a borate-phosphate buffer. In vitro protein solubility ($bf_1$_S, $bf_2$_S, $bf_3$_S) was measured in a borate-phosphate buffer with three different procedures. Results from laboratory techniques (in vitro) were correlated and linearly regressed with in sacco results. Of the three procedures of in vitro protein solubility evaluated, none of them could predict in sacco results with good precision. The highest Pearson correlation coefficient ($R^2$) was less than 0.50. Of two procedures of in vitro protein degradability studied, the $D_1$_Auf values were closely correlated with in sacco parameters: Kd, EDCP and %BCP with high R' values: 0.82, 0.85 and 0.85, respectively, and closely correlated with in sacco $D_j$ at 2, 4, 8 and 12 h rumen incubation with high $R^2$ values: 0.83, 0.91, 0.93 and 0.83, respectively. The $D_{24}$_Auf values could not predict in sacco results. The highest $R^2$ value was less then 0.40. These results indicated that in vitro protein solubility measured in borate-phosphate failed to identify differences in the rate and extent of protein degradation of legume seeds after dry roasting under various conditions and thus should not be used to predict rumen degradation, particularly for heat processed feedstuffs. But in vitro protein degradability using the modified Aufr re method by enzymatic hydrolysis for 1 h or possibly an intermediate time (>1 h and <24 h) is a promising laboratory procedure to detect effectiveness of dry roasting legume seeds on rumen protein degradation characteristics and could be used as a simple laboratory method to predict the rate and extent of protein degradation in the rumen in sacco with high accuracy. The equations to predict EDCP, Kd and BCP of dry roasted legume seeds (WLS and WFB) under various conditions are as follow: For both: EDCP (%)=-1.37+1.06*$D_1$_Auf ($R^2=0.85$, p<0.01). For both: Kd (%/h)=-21.81+0.49*$D_1$_Auf ($R^2=0.82$, p<0.01). For both: %BCP=103.37-1.07*$D_1$_Auf ($R^2=0.85$, p<0.01).

Effects of Surfactant Tween 80 on Forage Degradability and Microbial Growth on the In vitro Rumen Mixed and Pure Cultures

  • Goto, M.;Bae, H.;Lee, S.S.;Yahaya, M.S.;Karita, S.;Wanjae, K.;Cheng, K.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.5
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    • pp.672-676
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    • 2003
  • Effect of a surfactant Tween 80 on the bacterial growth in the rumen was examined on the in vitro pure cultures of Streptococcus bovis, Selenomonas ruminantium, Butyrivibrio fibrisolvens, Prevotella ruminicola, Megasphaera elsidenni, Fibrobacta succinogenes, Ruminanococcus albus and Ruminococcus flavefaciens. Dry matter degradability (DMD), concentrations and compositions of volatile fatty acids (VFA), and the most probable number (MPN) of cellulolytic bacteria and total number of bacteria in the presence of Tween 80 were also examined on the in vitro rumen mixed culture either with barley grain or orchardgrass hay. The growth of S. bovis, S. ruminantium, B. fibrisolvens, P. ruminicola, M. elsidenni and F. succinogenes were significantly higher (p<0.05) at over 0.05% concentrations of Tween 80 than those of the control cultures, while was not changed with R. albus and R. flavefaciens. With rumen mixed culture the DMD of barley grain and orchardgrass hay was significantly higher (p<0.05) at a 0.2% concentration of Tween 80 than the control, being reflected in the significantly higher (p<0.05) VFA production (mmol $g^{-1}$DDM) with orchardgrass hay. The higher (p<0.05) ratio of propionate to acetate at a 0.2% concentration of Tween 80 was also observed with orchardgrass hay, showing a similar trend with barley grain. No changes in the total bacterial number and MPN of cellulolytic bacteria were observed.

Manipulation of Rumen Fermentation by Yeast: The Effects of Dried Beer Yeast on the In vitro Degradability of Forages and Methane Production

  • Ando, S.;Khan, R.I.;Takahasi, J.;Gamo, Y.;Morikawa, R.;Nishiguchi, Y.;Hayasaka, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.1
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    • pp.68-72
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    • 2004
  • The effects of the addition of yeast on in vitro roughage degradability and methane production were investigated in order to clarify the effects of yeast on the rumen microbes and to establish methods of rumen manipulation. Three roughages (whole crop corn, rice straw and Italian ryegrass) were incubated for 3, 6, 12 and 24 h with or without dried beer yeast following the method described by Tilley and Terry. Using the same method, these roughages were incubated with or without yeast extract, albumin or purified DNA. In vitro methane production was measured with or without dried beer yeast at 12 and 24 h. The degradability of yeast was found to be 57 and 80% at 12 and 24 h, respectively. The rate of degradation of fraction b was 6.16%/h. There was a significant increase in roughage degradability at 6 h (p<0.05), 12 h (p<0.05) and 24 h (p<0.01) by dried yeast addition. The degradability of all three roughages was higher in the samples treated with yeast extract than in the no addition samples except in the case of rice straw incubated for 12 h. Nevertheless, the magnitude of increment was smaller with the addition of yeast extract than without the addition of yeast. With the addition of purified DNA, there were significant increases in roughage degradability at 6 h (p<0.01), 12 h (p<0.01) and 24 h (p<0.05); however, higher degradability values were detected in the samples to which albumin was added, particularly at 6 h. If the degradability values of the no addition samples with those of samples containing yeast, yeast extract, DNA and albumin were compared, the largest difference was found in the samples to which yeast was added, although it is worth noting that higher values were observed in the yeast extract samples than in the DNA or albumin samples, with the exception of the case of rice straw incubated for 24 h. Methane production was significantly increased at both 12 and 24 h incubation. The increment of roughage degradation and methane production brought about by the addition of dried beer yeast to the samples was thought to be due to the activation of rumen microbes. Water soluble fraction of yeast also seemed to play a role in ruminal microbe activation. The increment of degradability is thought to be partially due to the addition of crude protein or nucleic acid but it is expected that other factors play a greater role. And those factors may responsible for the different effects of individual yeast on ruminal microbes.

Ruminal Protein Degradation Characteristics of Cell Mass from Lysine Production

  • Seo, S.;Kim, H.J.;Lee, S.Y.;Ha, Jong K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.3
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    • pp.364-370
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    • 2008
  • Chemical analysis and in vitro studies were conducted to investigate the nutritive value for ruminants of cell mass from lysine production (CMLP) which is a by-product of the lysine manufacturing process. Proximate analysis, protein fractionation, and in vitro protein degradation using protease from Streptomyces griseus and strained ruminal fluid were carried out to estimate ruminal protein degradability of CMLP with two reference feedstuffs-soybean meal (SBM) and fish meal (FM). Amino acid composition and pepsin-HCl degradability were also determined to evaluate postruminal availability. CMLP contained 67.8% crude protein with a major portion being soluble form (45.4% CP) which was composed of mainly ammonium nitrogen (81.8% soluble CP). The amount of nucleic acids was low (1.15% DM). The total amount of amino acids contained in CMLP was 40.60% DM, which was lower than SBM (47.69% DM) or FM (54.08% DM). CMLP was composed of mainly fraction A and fraction B2, while the protein fraction in SBM was mostly B2 and FM contained high proportions of B2 and B3 fractions. The proportion of B3 fraction, slowly degradable protein, in CP was the highest in fish meal (23.34%), followed by CMLP (7.68%) and SBM (1.46%). CMLP was degraded up to 51.40% at 18 h of incubation with Streptomyces protease, which was low compared to FM (55.23%) and SBM (83.01%). This may be due to the insoluble portion of CMLP protein being hardly degradable by the protease. The in vitro fermentation by strained ruminal fluid showed that the amount of soluble fraction was larger in CMLP (40.6%) than in SBM (17.8%). However, because the degradation rate constant of the potentially degradable fraction of CMLP (2.0%/h) was lower than that of SBM (5.8%/h), the effective ruminal protein degradability of CMLP (46.95%) was slightly lower than SBM (53.77%). Unavailable fraction in the rumen was higher in CMLP (34.0%) compared to SBM (8.8%). In vitro CP degradability of CMLP by pepsin was 80.37%, which was lower than SBM (94.42%) and FM (89.04%). The evaluation of protein degradability using different approaches indicated that soluble protein in CMLP may supply a large amount of ammonia in the rumen while insoluble protein can be by-passed from microbial attacks due to its low degradability. The results from this study suggest that CMLP can be used as a protein supplement to ruminants for supplying both non-protein nitrogen to rumen microbes and rumen undegradable protein to the host animal.

Estimation of Ruminal Degradation and Intestinal Digestion of Tropical Protein Resources Using the Nylon Bag Technique and the Three-step In vitro Procedure in Dairy Cattle on Rice Straw Diets

  • Promkot, C.;Wanapat, Metha;Rowlinson, P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.12
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    • pp.1849-1857
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    • 2007
  • The experiment was carried out using fistulated multiparous Holstein Friesian crossbred (75% Holstein Friesian and 25% Red Sindhi) dairy cows in their dry period fed on untreated rice straw to evaluate the nutritive value of local protein feed resources using the in sacco method and in vitro pepsin-pancreatin digestion. Experimental feeds were cottonseed meal (CSM); soybean meal (SBM); dried brewery's grains (DBG); palm kernel meal (PSM); cassava hay (CH); leucaena leaf meal (LLM). Each feedstuff was weighed into duplicate nylon bags and incubated in each of the two rumen fistulated cows for 0, 2, 4, 8, 16, 24, and 48 h. Rumen feed residues from bags of 16 h incubation were used for estimation of lower gut digestibility by the technique of in vitro pepsin-pancreatin digestion. Ruminal ammonia-nitrogen ($NH_3-N$) concentrations did not differ between treatments or time with a mean of 5.5 mg%. Effective degradability of DM of CSM, SBM, DBG, PSM, CH and LLM were 41.9, 56.1, 30.8, 47.0, 41.1 and 47.5%, respectively. Effective degradabilities of the CP in feedstuffs were 49.6, 59.2, 40.9, 33.5, 47.3 and 65.0% for the respective feedstuffs. The CP in vitro pepsin-pancreatin digestibility as ranked from the highest to the lowest were SBM, CSM, LLM, CH, DBG, PSM, respectively. The intestinal and total tract digestion of feedstuffs in the current study were relatively lower than that obtained from previous literature. The results of this study indicate that SBM and LLM were highly degradable in the rumen, while CH, CSM and DBG were less degradable and, hence resulted in higher rumen undegradable protein. Soybean meal and LLM could be used to improve rumen ecology whilst CH, CSM and DBG could be used as rumen by-pass protein for ruminant feeding in the tropics.

Effects of L-glutamine supplementation on degradation rate and rumen fermentation characteristics in vitro

  • Suh, Jung-Keun;Nejad, Jalil Ghassemi;Lee, Yoon-Seok;Kong, Hong-Sik;Lee, Jae-Sung;Lee, Hong-Gu
    • Animal Bioscience
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    • v.35 no.3
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    • pp.422-433
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    • 2022
  • Objective: Two follow-up studies (exp. 1 and 2) were conducted to determine the effects of L-glutamine (L-Gln) supplementation on degradation and rumen fermentation characteristics in vitro. Methods: First, rumen liquor from three cannulated cows was used to test L-Gln (50 mM) degradation rate and ammonia-N production at 6, 12, 24, 36, and 48 h after incubation (exp. 1). Second, rumen liquor from two cannulated steers was used to assess the effects of five levels of L-Gln including 0% (control), 0.5%, 1%, 2%, and 3% at 0, 3, 6, 12, 24, 36, and 48 h after incubation on fermentation characteristics, gas production, and degradability of nutrients (exp. 2). Results: In exp. 1, L-Gln degradation rate and ammonia-N concentrations increased over time (p<0.001). In exp. 2, pH was reduced significantly as incubation time elapsed (p<0.001). Total gas production tended to increase in all groups as incubation time increased. Acetate and propionate tended to increase by increasing glutamine (Gln) levels, whereas levels of total volatile fatty acids (VFAs) were the highest in 0.5% and 3% Gln groups (p<0.001). The branched-chain VFA showed both linear and quadratic effects showing the lowest values in the 1% Gln group particularly after 6 h incubation (p<0.001). L-Gln increased crude protein degradability (p<0.001), showing the highest degradability in the 0.5% Gln group regardless of incubation time (p<0.05). Degradability of acid detergent fiber and neutral detergent fiber showed a similar pattern showing the highest values in 0.5% Gln group (p<0.10). Conclusion: Although L-Gln showed no toxicity when it was supplemented at high dosages (2% to 3% of DM), 0.5% L-Gln demonstrated the positive effects on main factors including VFAs production in-vitro. The results of this study need to be verified in further in-vivo study.