• Title/Summary/Keyword: Rumen Buffer

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Distribution and Activities of Hydrolytic Enzymes in the Rumen Compartments of Hereford Bulls Fed Alfalfa Based Diet

  • Lee, S.S.;Kim, C.-H.;Ha, J.K.;Moon, Y.H.;Choi, N.J.;Cheng, K.-J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.12
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    • pp.1725-1731
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    • 2002
  • The distribution and activities of hydrolytic enzymes (cellulolyti, hemicellulolytic,pectinolytic and others) in the rumen compartments of Hereford bulls fed 100% alfalfa hay based diets were evaluated. The alfalfa proportion in the diet was gradually increased for two weeks. Whole rumen contents were processed into four fractions: Rumen contents including both the liquid and solid fractions were homogenized and centrifuged, and the supernatant was assayed for enzymes located in whole rumen contents (WRE); rumen contents were centrifuged and the supernatant was assayed for enzymes located in rumen fluids (RFE); feed particles in rumen contents were separated manually, washed with buffer, resuspended in an equal volume of buffer, homogenized and centrifuged and supernatant was assayed for enzymes associated with feed particles (FAE); and rumen microbial cell fraction was separated by centrifugation, suspended in an equal volume of buffer, sonicated and centrifuged, and the supernatant was assayed for enzymes bound with microbial cells (CBE). It was found that polysaccharide-degrading proteins such as $\beta$-1,4-D-endoglucanase, $\beta$-1,4-D-exoglucanase, xylanase and pectinase enzymes were located mainly with the cell bound (CBE) fraction. However, $\beta$-D-glucosidase, $\beta$-D-fucosidase, acetylesterase, and $\alpha$-L-arabinofuranosidase were located in the rumen fluids (RFE) fraction. Protease activity distributions were 37.7, 22.1 and 40.2%, and amylase activity distributions were 51.6, 18.2 and 30.2% for the RFE, FAE and CBE fractions, respectively. These results indicated that protease is located mainly in rumen fluid and with microbial cells, whereas amylase was located mainly in the rumen fluid.

Evaluation of glycerol encapsulated with alginate and alginate-chitosan polymers in gut environment and its resistance to rumen microbial degradation

  • Gawad, Ramadan;Fellner, Vivek
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.1
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    • pp.72-81
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    • 2019
  • Objective: To determine the effect of gut pH and rumen microbial fermentation on glycerol encapsulated in alginate and alginate-chitosan polymers. Methods: Glycerol was encapsulated at 2.5%, 5%, 7.5%, or 10% (w/w) with sodium alginate (A) and alginate-chitosan (AC) polymers. Surface morphology and chemical modifications of the beads were evaluated using scanning electron microscopy and Fourier transform infrared (FTIR) spectra. Encapsulation efficiency was determined at the 5% glycerol inclusion level in two experiments. In experiment 1, 0.5 g of alginate-glycerol (AG) and alginate-chitosan glycerol (ACG) beads were incubated for 2 h at $39^{\circ}C$ in pH 2 buffer followed by 24 h in pH 8 buffer to simulate gastric and intestinal conditions, respectively. In experiment 2, 0.5 g of AG and ACG beads were incubated in pH 6 buffer at $39^{\circ}C$ for 8 h to simulate rumen conditions. All incubations were replicated four times. Free glycerol content was determined using a spectrophotometer and used to assess loading capacity and encapsulation efficiency. An in vitro experiment with mixed cultures of rumen microbes was conducted to determine effect of encapsulation on microbial fermentation. Data were analyzed according to a complete block design using the MIXED procedure of SAS (SAS Institute, Cary, NC, USA). Results: For AG and ACG, loading capacity and efficiency were 64.7%, 74.7%, 70.3%, and 78.1%, respectively. Based on the FTIR spectra and scanning electron microscopy, ACG treatment demonstrated more intense and stronger ionic bonds. At pH 6, 36.1% and 29.7% of glycerol was released from AG and ACG, respectively. At pH 2 minimal glycerol was released but pH 8 resulted in 95.7% and 93.9% of glycerol released from AG and ACG, respectively. In vitro microbial data show reduced (p<0.05) fermentation of encapsulated glycerol after 24 h of incubation. Conclusion: The AC polymer provided greater protection in acidic pH with a gradual release of intact glycerol when exposed to an alkaline pH.

Using Plant Source as a Buffering Agent to Manipulating Rumen Fermentation in an In vitro Gas Production System

  • Kang, S.;Wanapat, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.10
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    • pp.1424-1436
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    • 2013
  • The objective of this study was to investigate the effect of banana flower powder (BAFLOP) supplementation on gas production kinetics and rumen fermentation efficiency in in vitro incubation with different ratios of roughage to concentrate in swamp buffalo and cattle rumen fluid. Two male, rumen fistulated dairy steers and swamp buffaloes were used as rumen fluid donors. The treatments were arranged according to a $2{\times}2{\times}3$ factorial arrangement in a Completely randomized design by using two ratios of roughage to concentrate (R:C; 75:25 and 25:75) and 3 levels of BAFLOP supplementation (0, 2 and 4% of dietary substrate) into two different kinds of rumen fluid (beef cattle and swamp buffalo). Under this investigation, the results revealed that the rumen ecology was affected by R:C ratio. The pH declined as a result of using high concentrate ratio; however, supplementation of BAFLOP could buffer the pH which led to an improvement of ruminal efficiency. BAFLOP supplementation affected acetic acid (C2) when the proportion of concentrate was increased. However, there were no effect on total volatile fatty acid (TVFA) and butyric acid (C4) by BAFLOP supplementation. The microbial community was affected by BAFLOP supplementation, especially the bacterial population. As revealed by real-time PCR, the populations of F. succinogenes and R. albus were reduced by the high concentrate treatments while that of R. flavafaciens were increased. The populations of three dominant cellulolytic bacteria were enhanced by BAFLOP supplementation, especially on high concentrate diet. BAFLOP supplementation did not influence the ammonia nitrogen ($NH_3$-N) concentration, while R:C did. In addition, the in vitro digestibility was improved by either R:C or BAFLOP supplementation. The BAFLOP supplementation showed an effect on gas production kinetics, except for the gas production rate constant for the insoluble fraction (c), while treatments with high concentrate ratio resulted in the highest values. In addition, BAFLOP tended to increase gas production. Based on this study, it could be concluded that R:C had an effect on rumen ecology both in buffalo and cattle rumen fluid and hence, BAFLOP could be used as a rumen buffering agent for enhancing rumen ecology fed on high concentrate diet. It is recommended that level of BAFLOP supplementation should be at 2 to 4% of total dry matter of substrate. However, in vivo trials should be subsequently conducted to investigate the effect of BAFLOP in high concentrate diets on rumen ecology as well as ruminant production.

Effects of Rumen pH on Degradation Kinetics and Fermentation Indices of Corn Silage Ensiled with Antifungal and Carboxylesterase Producing Inoculants

  • Chang, Hong Hee;Paradhipta, Dimas Hand Vidya;Lee, Seong Shin;Lee, Hyuk Jun;Joo, Young Ho;Min, Hyeong Gyu;Kim, Sam Churl
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.40 no.3
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    • pp.131-137
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    • 2020
  • The present study investigated effects of antifungal and carboxylesterase inoculant on rumen fermentation with different rumen pH. Corn silage was treated without inoculant (CON) and with a mixed Lactobacillus brevis 5M2 and L. buchneri 6M1 (MIX). Rumen fluid was collected from two cannulated Hanwoo heifers before morning feeding (high rumen pH at 6.70) and 3 h after feeding (low rumen pH at 6.20). Dried corn silage was incubated in the rumen buffer (rumen fluid + anaerobic culture medium at 1:2 ratio) for 48 h at 39℃. Eight replications for each treatment were used along with two blanks. Both in a high and a low rumen pH, MIX silages presented higher (p<0.05) the immediately degradable fraction, the potentially degradable fraction, total degradable fraction, and total volatile fatty acid (VFA) than those of CON silages. Incubated corn silages in a low rumen pH presented lower (p<0.05) total degradable fraction, ammonia-N, total VFA (p=0.061), and other VFA profiles except acetate and propionate, than those in a high rumen pH. The present study concluded that application of antifungal and carboxylesterase inoculant on corn silage could improve degradation kinetics and fermentation indices in the rumen with high and low pH conditions.

Effect of C18-polyunsaturated Fatty Acids on Their Direct Incorporation into the Rumen Bacterial Lipids and CLA Production In vitro

  • Choi, S.H.;Song, M.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.4
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    • pp.512-515
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    • 2005
  • An in vitro study was conducted to determine the effect of C18-polyunsaturated fatty acid on direct incorporation into the rumen bacteria, bio-hydrogenation and production of CLA in vitro. Sixty milligrams of linoleic acid ($C_{18:2}$) or linolenic acid ($C_{18:3}$) were absorbed into the 0.5 g cellulose powder was added to the 150 ml culture solution consisting of 120 ml McDougall's buffer and 30 ml strained rumen fluid. Four uCi of 1-$^{14}C_{18:2}$ or 1-$^{14}C_{18:3}$ (1 uCi/15 mg each fatty acid) were also added to the corresponding fatty acids to estimate the direct incorporation into the bacterial lipids. The culture solution was then incubated anaerobically in a culture jar with stirrer at 39$^{\circ}C$ for 12 h. Ammonia concentration and pH of the culture solution were slightly influenced by the fatty acids. Amount of fatty acid incorporated into the bacteria was 1.20 mg and 0.43 mg/30 ml rumen fluid for $C_{18:2}$ and $C_{18:3}$, respectively during 12 h incubation. Slightly increased CLA (sum of cis-9, trans-11 and cis-10, trans-12 $C_{18:2}$) was obtained from the $C_{18:3}$ addition compared to that from $C_{18:2}$ after 12 h incubation in vitro.

Effects of rumen-protected amino acid prototypes on rumen fermentation characteristics in vitro

  • Gyeongjin, Kim;Tabita Dameria, Marbun;Jinhyun, Park;Sang Moo, Lee;Hong Gu, Lee;Jun Ok, Moon;Jin Seung, Park;Eun Joong, Kim
    • Korean Journal of Agricultural Science
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    • v.48 no.4
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    • pp.669-679
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    • 2021
  • This study was conducted to evaluate the effects of rumen-protected amino acid (RPAA) prototypes, which were chemically synthesized, on in vitro rumen fermentation and protection rate outcomes. Several RPAA prototypes were incubated with timothy hay and concentrate. Treatments consisted of 1) control (CON; no RPAA prototype supplement), and prototypes of 2) 0.5% RP-methionine (RPMet), 3) 0.5% RP-tryptophan (RPTrp), 4) 0.5% RP-valine (RPVal), 5) 0.5% RP-phenylalanine (RPPhe), 6) 0.5% RP-leucine (RPLeu), 7) 0.5% RP-histidine (RPHis), 8) 20% RPMet, and 9) 20% RPTrp (w·w-1 feed). The inoculum (50 mL) prepared with rumen fluid and McDougall's buffer (1 : 4) was dispensed in individual serum bottles and was anaerobically incubated for 0, 6, and 24 h at 39℃ in triplicate. The dry matter degradability did not differ among the groups, except for the 20% RPMet and the 20% RPTrp treatments at 6 and 24 h. The total volatile fatty acid concentration in the 20% RPMet was higher (p < 0.05) than the rest of the groups at 6 h, and 20% RPMet showed the highest molar proportion of acetate, whereas the lowest proportion of propionate was found at 6 h (p < 0.05). The protection rate of the RPAA prototypes ranged from 29.85 to 109.21%. at 24 h. In conclusion, the chemically synthesized RPAA prototypes studied here had no detrimental effects on rumen fermentation parameters. Further studies using animal models are needed for more accurate evaluations of the effectiveness of RPAA.

Potency of cashew nut shell liquid in rumen modulation under different dietary conditions and indication of its surfactant action against rumen bacteria

  • Oh, Seongjin;Suzuki, Yasuyuki;Hayashi, Shusuke;Suzuki, Yutaka;Koike, Satoshi;Kobayashi, Yasuo
    • Journal of Animal Science and Technology
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    • v.59 no.11
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    • pp.27.1-27.7
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    • 2017
  • Background: Cashew nut shell liquid (CNSL) is an agricultural byproduct containing alkylphenols that has been shown to favorably change the rumen fermentation pattern only under experimentally fixed feeding conditions. Investigation of CNSL potency in rumen modulation under a variety of feeding regimens, and evidence leading to the understanding of CNSL action are obviously necessary for further CNSL applications. The objective of this study was to evaluate the potency of CNSL for rumen modulation under different dietary conditions, and to visually demonstrate its surfactant action against selected rumen bacteria. Methods: Batch culture studies were carried out using various diets with 5 different forage to concentrate (F:C) ratios (9:1, 7:3, 5:5. 3:7 and 1:9). Strained rumen fluid was diluted with a buffer and incubated with each diet. Gas and short chain fatty acid (SCFA) profiles were characterized after 18 h incubation at $39^{\circ}C$. Monensin was also evaluated as a reference additive under the same conditions. Four species of rumen bacteria were grown in pure culture and exposed to CNSL to determine their morphological sensitivity to the surfactant action of CNSL. Results: CNSL supplementation decreased total gas production in diets with 5:5 and 3:7 F:C ratios, whereas the F:C ratio alone did not affect any gas production. Methane decrease by CNSL addition was more apparent in diets with 5:5, 3:7, and 1:9 F:C ratios. An interactive effect of CNSL and the F:C ratio was also observed for methane production. CNSL supplementation enhanced propionate production, while total SCFA production was not affected. Monensin decreased methane production but only in a diet with a 1:9 F:C ratio with increased propionate. Studies of pure cultures indicated that CNSL damaged the cell surface of hydrogen- and formate-producing bacteria, but did not change that of propionate-producing bacteria. Conclusion: CNSL can selectively inhibit rumen bacteria through its surfactant action to lead fermentation toward less methane and more propionate production. As CNSL is effective over a wider range of dietary conditions for such modulation of rumen fermentation in comparison with monensin, this new additive candidate might be applied to ruminant animals for various production purposes and at various stages.

The Effect of Castor Aralia (Kalopanax pictus Nakai) Trunk Extracts on Rumen Fermentation and Methane Reduction In vitro (엄나무 (Kalopanax pictus Nakai) 줄기 추출물이 In vitro 반추위 발효와 메탄저감에 미치는 영향)

  • Kim, Jae Seong;Hwang, Moon Seok;Kim, Yong Chae;Yoon, Young-Man;Bae, Gui Sek;Kim, Chang-Hyun
    • Journal of Animal Environmental Science
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    • v.21 no.3
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    • pp.113-122
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    • 2015
  • An experiment was conducted to examine the effects of Kalopanax pictus Nakai (Kalopanax) on in vitro rumen fermentation and methane (CH4) reduction. Kalopanax trunk was extracted with 70% ethanol and 70% methanol. Rumen fluid, alfalfa hay and buffer (control: C) supplemented with 0.3% Kalopanx juice (T1), 0.3% ethanol extract (T2) and 0.3% methanol extract (T3) in the total volume of culture medium were incubated at $38^{\circ}C$ for 24h and 48h. Rumen pH was lower in all Kalopanax treatments during all incubations than that in control (p<0.05). Total VFA and total gas production in T2 and T3 was significantly higher than that in C at 48h incubation (p<0.05). Ammonia-N was decreased in all treatments compared with C during the incubation periods (p<0.05). At 24h incubation, $CH_4$ contents were significantly reduced by both alcohol extracts. It is concluded that supplementing Kalopanax extracts can stimulate ruminal fermentation of rumen microorganisms and inhibit methanogenesis.

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).

Improving Nutritional Quality of Cocoa Pod (Theobroma cacao) through Chemical and Biological Treatments for Ruminant Feeding: In vitro and In vivo Evaluation

  • Laconi, Erika B.;Jayanegara, Anuraga
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.3
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    • pp.343-350
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    • 2015
  • Cocoa pod is among the by-products of cocoa (Theobroma cacao) plantations. The aim of this study was to apply a number of treatments in order to improve nutritional quality of cocoa pod for feeding of ruminants. Cocoa pod was subjected to different treatments, i.e. C (cocoa pod without any treatment or control), CAm (cocoa pod+1.5% urea), CMo (cocoa pod+3% molasses), CRu (cocoa pod+3% rumen content) and CPh (cocoa pod+3% molasses+Phanerochaete chrysosporium inoculum). Analysis of proximate and Van Soest's fiber fraction were performed on the respective treatments. The pods were then subjected to an in vitro digestibility evaluation by incubation in rumen fluid-buffer medium, employing a randomized complete block design (n = 3 replicates). Further, an in vivo evaluation of the pods (35% inclusion level in total mixed ration) was conducted by feeding to young Holstein steers (average body weight of $145{\pm}3.6kg$) with a $5{\times}5$ latin square design arrangement (n = 5 replicates). Each experimental period lasted for 30 d; the first 20 d was for feed adaptation, the next 3 d was for sampling of rumen liquid, and the last 7 d was for measurements of digestibility and N balance. Results revealed that lignin content was reduced significantly when cocoa pod was treated with urea, molasses, rumen content or P. chrysosporium (p<0.01) with the following order of effectiveness: CPh>CAm>CRu>CMo. Among all treatments, CAm and CPh treatments significantly improved the in vitro dry matter and organic matter digestibility (p<0.05) of cocoa pod. Average daily gain of steers receiving CAm or CPh treatment was significantly higher than that of control (p<0.01) with an increase of 105% and 92%, respectively. Such higher daily gain was concomitant with higher N retention and proportion of N retention to N intake in CAm and CPh treatments than those of control (p<0.05). It can be concluded from this study that treatment with either urea or P. chrysosporium is effective in improving the nutritive value of cocoa pod.