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Sample records for rumen degradable protein

  1. Effects of Synchronicity of Carbohydrate and Protein Degradation on Rumen Fermentation Characteristics and Microbial Protein Synthesis

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    J. K. Seo

    2013-03-01

    Full Text Available A series of in vitro studies were carried out to determine i the effects of enzyme and formaldehyde treatment on the degradation characteristics of carbohydrate and protein sources and on the synchronicity of these processes, and ii the effects of synchronizing carbohydrate and protein supply on rumen fermentation and microbial protein synthesis (MPS in in vitro experiments. Untreated corn (C and enzyme-treated corn (EC were combined with soy bean meal with (ES and without (S enzyme treatment or formaldehyde treatment (FS. Six experimental feeds (CS, CES, CFS, ECS, ECES and ECFS with different synchrony indices were prepared. Highly synchronous diets had the greatest dry matter (DM digestibility when untreated corn was used. However, the degree of synchronicity did not influence DM digestibility when EC was mixed with various soybean meals. At time points of 12 h and 24 h of incubation, EC-containing diets showed lower ammonia-N concentrations than those of C-containing diets, irrespective of the degree of synchronicity, indicating that more efficient utilization of ammonia-N for MPS was achieved by ruminal microorganisms when EC was offered as a carbohydrate source. Within C-containing treatments, the purine base concentration increased as the diets were more synchronized. This effect was not observed when EC was offered. There were significant effects on VFA concentration of both C and S treatments and their interactions. Similar to purine concentrations, total VFA production and individual VFA concentration in the groups containing EC as an energy source was higher than those of other groups (CS, CES and CFS. The results of the present study suggested that the availability of energy or the protein source are the most limiting factors for rumen fermentation and MPS, rather than the degree of synchronicity.

  2. Effects of grain source, grain processing, and protein degradability on rumen kinetics and microbial protein synthesis in Boer kids.

    Science.gov (United States)

    Brassard, M-E; Chouinard, P Y; Berthiaume, R; Tremblay, G F; Gervais, R; Martineau, R; Cinq-Mars, D

    2015-11-01

    Microbial protein synthesis in the rumen would be optimized when dietary carbohydrates and proteins have synchronized rates and extent of degradation. The aim of this study was to evaluate the effect of varying ruminal degradation rate of energy and nitrogen sources on intake, nitrogen balance, microbial protein yield, and kinetics of nutrients in the rumen of growing kids. Eight Boer goats (38.2 ± 3.0 kg) were used. The treatments were arranged in a split-plot Latin square design with grain sources (barley or corn) forming the main plots (squares). Grain processing methods and levels of protein degradability formed the subplots in a 2 × 2 factorial arrangement for a total of 8 dietary treatments. The grain processing method was rolling for barley and cracking for corn. Levels of protein degradability were obtained by feeding untreated soybean meal (SBM) or heat-treated soybean meal (HSBM). Each experimental period lasted 21 d, consisting of a 10-d adaptation period, a 7-d digestibility determination period, and a 4-d rumen evacuation and sampling period. Kids fed with corn had higher purine derivatives (PD) excretion when coupled with SBM compared with HSBM and the opposite occurred with barley-fed kids ( ≤ 0.01). Unprocessed grain offered with SBM led to higher PD excretion than with HSBM whereas protein degradability had no effect when processed grain was fed ( ≤ 0.03). Results of the current experiment with high-concentrate diets showed that microbial N synthesis could be maximized in goat kids by combining slowly fermented grains (corn or unprocessed grains) with a highly degradable protein supplement (SBM). With barley, a more rapidly fermented grain, a greater microbial N synthesis was observed when supplementing a low-degradable protein (HSBM).

  3. In vitro estimation of rumen protein degradability using 35S to label the bacterial mass

    International Nuclear Information System (INIS)

    Khristov, A.; Aleksandrov, S.; Aleksiev, I.

    1994-01-01

    An experiment was carried out in order to simplify a previously developed 15 N-method for in vitro estimation of rumen protein degradability. Casein (Cas), whole soybeans (Sb) heated at 120 o C for 20 min (SbTherm) and sunflower (Sfl) were incubated at 39 o C for 4 hours in a water bathshaker with the following media: McDougall's buffer, strained and enriched with particle associated bacteria rumen fluid (2:1), rapidly (maltose, sucrose, glucose) and more slowly (pectin, soluble starch) degradable carbohydrates with final concentration of 815 mg/100 ml and 21.7 μCi/100 ml of 35 S (from Na 2 35 SO 4 ). After the incubation had been ceased, a bacterial fraction was isolated through differential centrifugation and specific activity of bacterial (Bac) and high speed total solids (TS) nitrogen was measured. The ratio was used to calculate bacterial mass in TS and through the Kjeldahl nitrogen concentration in TS - the net bacterial growth (against control vessels without protein). The level of ammonia-N in the supernate after blank correction was used to find the ammonia-N released from protein degradation. The data showed that the rate (and extend) of degradation for the Cas (as a standard protein) was lower compared to those obtained through the 15 N-method but it was higher than the rate derived through another in vitro method. The Cas equivalent of the Sb was higher than the figure we found in a previous experiment with solvent extracted soybean meal suggesting that the 35 S-method underestimated the degradability of the Cas. After being tested on a wider range of foodstuffs, the proposed 35 S-method might be considered as an alternative procedure which is less laborous than the 15 N-method. (author)

  4. The effects of high levels of rumen degradable protein on rumen pH and histamine concentrations in dairy cows

    NARCIS (Netherlands)

    Pilachai, R.; Schonewille, J.T.; Thamrongyoswittayakul, C.; Aiumlamai, S.; Wachirapakom, C.; Everts, H.; Hendriks, W.H.

    2012-01-01

    An experiment was conducted to test the hypothesis that the supplementation of crude protein (CP) results in rumen acidosis and increased histamine concentrations in dairy cows. Six ruminally fistulated, non-pregnant dry cows were fed three experimental rations in a replicated 3 × 3 Latin square

  5. Estimation of Relationship Between In Situ and In Vitro Rumen Protein Degradability of Extruded Full Fat Soybean

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    Arzu Erol Tunç

    2017-10-01

    Full Text Available The objectives of this study were to estimate the protein degradability of extruded full fat soybean (ESB by in situ (nylon bag and in vitro enzymatic method and to develop an equation in order predict in situ degradability from in vitro values. In the study enzymatic technique; hydrolysis after 1 h (INV1 and after 24 h (INV24 by a purified protease extracted from Streptomyces griseus in a borate-phosphate buffer at pH 8 was used as in vitro method. Relationship between in situ effective protein degradability (INSE and in vitro degradability after 1 and 24 hours incubations (INV1 and INV24 were determined. In situ protein degradability was measured at 0, 2, 4, 8, 16, 24, and 48 and at 72 h incubations in the rumen of 3 Holstein cows. In the study INSE, INV1 and INV24 were determined as 58.05, 20.24 and 41.46% respectively. Despite there were differences between in situ and in vitro protein degradability values, correlation coefficients between in situ and in vitro protein degradability of ESB were high and regression equations for estimation of in situ from in vitro were found significant. As conclusion in vitro enzymatic protein degradability (INV1 and INV24 can be used for estimation of in situ effective protein degradability of extruded full fat soybean.

  6. Rumen Degradability and Post-ruminal Digestion of Dry Matter, Nitrogen and Amino Acids of Three Protein Supplements

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    Wei Gao

    2015-04-01

    Full Text Available This study evaluated the in situ ruminal degradability, and subsequent small intestinal digestibility (SID of dry matter, crude protein (CP, and amino acids (AA of cottonseed meal (CSM, sunflower seed meal (SFSM and distillers dried grains with solubles (DDGS by using the modified three-step in vitro procedure. The ruminal degradability and subsequent SID of AA in rumen-undegradable protein (RUP-AA varied among three protein supplements. The result show that the effective degradability of DM for SFSM, CSM, and DDGS was 60.8%, 56.4%, and 41.0% and their ruminal fermentable organic matter was 60.0%, 55.9%, and 39.9%, respectively. The ruminal degradable protein (RDP content in CP for SFSM, CSM, and DDGS was 68.3%, 39.0%, and 32.9%, respectively, at the ruminal solid passage rate of 1.84%/h. The SFSM is a good source of RDP for rumen micro-organisms; however, the SID of RUP of SFSM was lower. The DDGS and CSM are good sources of RUP for lambs to digest in the small intestine to complement ruminal microbial AA of growing lambs. Individual RUP-AA from each protein source was selectively removed by the rumen micro-organisms, especially for Trp, Arg, His, and Lys (p<0.01. The SID of individual RUP-AA was different within specific RUP origin (p<0.01. Limiting amino acid was Leu for RUP of CSM and Lys for both RUP of SFSM and DDGS, respectively. Therefore, different protein supplements with specific limitations should be selected and combined carefully in growing lambs ration to optimize AA balance.

  7. Rumen Degradability and Post-ruminal Digestion of Dry Matter, Nitrogen and Amino Acids of Three Protein Supplements.

    Science.gov (United States)

    Gao, Wei; Chen, Aodong; Zhang, Bowen; Kong, Ping; Liu, Chenli; Zhao, Jie

    2015-04-01

    This study evaluated the in situ ruminal degradability, and subsequent small intestinal digestibility (SID) of dry matter, crude protein (CP), and amino acids (AA) of cottonseed meal (CSM), sunflower seed meal (SFSM) and distillers dried grains with solubles (DDGS) by using the modified three-step in vitro procedure. The ruminal degradability and subsequent SID of AA in rumen-undegradable protein (RUP-AA) varied among three protein supplements. The result show that the effective degradability of DM for SFSM, CSM, and DDGS was 60.8%, 56.4%, and 41.0% and their ruminal fermentable organic matter was 60.0%, 55.9%, and 39.9%, respectively. The ruminal degradable protein (RDP) content in CP for SFSM, CSM, and DDGS was 68.3%, 39.0%, and 32.9%, respectively, at the ruminal solid passage rate of 1.84%/h. The SFSM is a good source of RDP for rumen micro-organisms; however, the SID of RUP of SFSM was lower. The DDGS and CSM are good sources of RUP for lambs to digest in the small intestine to complement ruminal microbial AA of growing lambs. Individual RUP-AA from each protein source was selectively removed by the rumen micro-organisms, especially for Trp, Arg, His, and Lys (p<0.01). The SID of individual RUP-AA was different within specific RUP origin (p<0.01). Limiting amino acid was Leu for RUP of CSM and Lys for both RUP of SFSM and DDGS, respectively. Therefore, different protein supplements with specific limitations should be selected and combined carefully in growing lambs ration to optimize AA balance.

  8. Effects of rumen-degradable protein:rumen-undegradable protein ratio and corn processing on production performance, nitrogen efficiency, and feeding behavior of Holstein dairy cows.

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    Savari, M; Khorvash, M; Amanlou, H; Ghorbani, G R; Ghasemi, E; Mirzaei, M

    2018-02-01

    This study was conducted to investigate the effects of the ratio of rumen-degradable protein (RDP) to rumen-undegradable protein (RUP) and corn processing method on production performance, nitrogen (N) efficiency, and feeding behavior of high-producing Holstein dairy cows. Twelve multiparous Holstein cows (second parity; milk yield = 48 ± 3 kg/d) were assigned to a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Factor 1 was corn processing method [ground corn (GC) or steam flaked corn (SFC) with a flake density of about 390 g/L], and factor 2 was RDP:RUP ratio [low ratio (LR) = 60:40; high ratio (HR) = 65:35] based on crude protein (%). The crude protein concentrations were kept constant across the treatments (16.7% of DM). No significant interactions of main treatment effects occurred for lactation performance data. Cows fed 2 different RDP:RUP ratios exhibited similar dry matter intake (DMI), but those fed SFC showed decreased feed intake compared with those receiving GC (25.1 ± 0.48 vs. 26.2 ± 0.47 kg/d, respectively). Cows fed HR diets produced more milk than did those fed LR diets (44.4 ± 1.05 vs. 43.2 ± 1.05 kg/d, respectively). Milk fat content decreased but milk protein content increased in cows fed SFC compared with those fed GC. Feed efficiency (i.e., milk yield/DMI) was enhanced with increasing ratio of RDP:RUP (1.68 ± 0.04 vs. 1.74 ± 0.04 for LR and HR, respectively). Apparent N efficiency was higher in cows fed HR than in those fed LR (30.4 ± 0.61 vs. 29.2 ± 0.62, respectively). Compared with cows fed the GC-based diet, those receiving SFC exhibited lower values of N intake, N-NH 3 concentration, and fecal N excretion. Cows receiving SFC-based diets spent more time ruminating (min/kg of DMI) than did those fed GC. Although these results showed no interaction effects of RDP:RUP ratio and corn processing method on performance, higher RDP:RUP ratios and ground corn can be effective feeding strategies for

  9. Lowering rumen-degradable and rumen-undegradable protein improved amino acid metabolism and energy utilization in lactating dairy cows exposed to heat stress.

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    Kaufman, J D; Pohler, K G; Mulliniks, J T; Ríus, A G

    2018-01-01

    The objective of this study was to evaluate the effects of reducing dietary rumen-degradable protein (RDP) and rumen-undegradable protein (RUP) on protein and energy metabolism in heat-stressed dairy cows. Eighteen primiparous and 30 multiparous mid-lactation Holstein cows were used in a completely randomized design arranged in a 2 × 2 factorial (n = 12/treatment). Cows were randomly assigned to 1 of 4 dietary treatments that included 2 levels of RDP (10 and 8%; D) and 2 levels of RUP (8 and 6%; U) of dry matter for 21 d as (1) 10D:8U, (2) 8D:8U, (3) 10D:6U, and (4) 8D:6U. Diets were isoenergetic and contained 50% forage and 50% concentrate (dry matter basis). Cows were housed in a freestall barn. Three weeks before start of treatments, all animals were fed the 10D:8U diet and received supplemental cooling to prevent heat stress. During the treatment period, cows experienced a daily increment in temperature-humidity index from 74 to 82 for 1000 to 2000 h. Blood samples were collected on d -1 and 21 of the treatment period to determine plasma concentrations of AA, glucose, insulin, fatty acids, and β-hydroxybutyrate. For primiparous cows, reducing from 10 to 8% RDP decreased insulin concentrations. For multiparous cows, we found significant RDP by RUP interactions for insulin, β-hydroxybutyrate, fatty acids, total essential AA, and 3-methylhistidine concentrations. Reducing from 10 to 8% RDP decreased insulin concentrations at 6% RUP, but concentrations did not change when reducing RDP at 8% RUP. Reducing from 10 to 8% RDP decreased β-hydroxybutyrate concentrations at 8% RUP, but concentrations did not change when reducing RDP at 6% RUP. Reducing from 10 to 8% RDP increased nonesterified fatty acid and total essential AA concentrations at 8% RUP, but concentrations did not change when reducing RDP at 6% RUP. Reducing from 8 to 6% RUP decreased 3-methylhistidine concentration at 8% RDP, but not at 10% RDP. Reducing from 8 to 6% RUP increased milk protein yield

  10. Effects of molasses and corn grain at 2 levels of ruminally degradable protein on lactating cow ruminal fermentation and rumen content mass

    Science.gov (United States)

    The objective of this study was to evaluate lactating dairy cow ruminal fermentation and rumen content mass with diets containing molasses (M) or finely ground dry corn grain at 3 levels of M (0, 5.25, 10.5% DM) and with differing levels of ruminally degradable protein (+RDP or –RDP). Twelve ruminal...

  11. Influence of rumen protein degradability on productive and reproductive performance in buffalo cows.

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    Campanile, Giuseppe; Di Palo, Rossella; Infascelli, Federico; Gasparrini, Bianca; Neglia, Gianluca; Zicarelli, Fabio; D'Occhio, Michael J

    2003-01-01

    The present study aimed to ascertain the influence of crude protein (CP) digestibility in the rumen on the quantity and quality of milk production and reproductive performance, blood (BU) and milk (MU) urea, haematological profile and vaginal mucus urea, ammonia and potassium of buffalo cows. Lactating buffaloes (n = 84), 60 days in milk, were randomly subdivided into Group C (control, n = 42) and Group T (fed a diet supplemented with Aspergillus oryzae, n = 42). In three fistulated buffalo, the diet supplemented with Aspergillus oryzae showed a decrease (P milk, were used to study the haematological profile and to determine milk urea and ammonia in the vaginal mucus. The animals in Group T1 had higher ammonia values in the blood (P mucus than Group C1. A relationship was found between MU and BU. MU was influenced by CP intake and dry matter intake. No differences between the treatments were observed in reproductive performance and the conception rate and calving interval were 37.9% and 41.4% (90 trial-day) and 449 and 419 days respectively in Groups T and C. Reproductive performance was not influenced by high levels of BU nor by blood ammonia levels, although the latter were higher in the group fed the diet supplemented with Aspergillus oryzae.

  12. The use of a tannin crude extract from Cistus ladanifer L. to protect soya-bean protein from degradation in the rumen.

    Science.gov (United States)

    Dentinho, M T P; Moreira, O C; Pereira, M S; Bessa, R J B

    2007-06-01

    Cistus ladanifer L. (CL) is a perennial shrub abundant in dry woods and dry land of Mediterranean zone, with high level of tannins. Tannins bind to protein, preventing its degradation in the digestive compartments. This tannin/protein complex may be advantageous when partially protecting good-quality feed protein from excessive rumen protein degradation. The objective of this trial was to use a CL phenol crude extract to prevent excessive rumen degradation of soya-bean meal protein. The phenolic compounds were extracted using an acetone/water solution (70:30, v/v). Soya-bean meal was then treated with this crude CL extract, containing 640 g of total phenols (TP) per kg of dry matter (DM), in order to obtain mixtures with 0, 12.5, 25, 50, 100 and 150 g of TP per kg DM. Three rumen-cannulated rams were used to assess in sacco rumen degradability of DM and nitrogen (N). The three-step in vitro procedure was used to determine intestinal digestibility. Increasing extract concentrations quadratically decreased the N-soluble fraction a (R2 = 0.96, P = 0.0001) and increased the non-soluble degradable fraction b (R2 = 0.92, P = 0.005). The rate of degradation c linearly decreased with CL extract doses (R2 = 0.44, P = 0.0065). For the effective rumen degradability of N, a linear reduction (R2 = 0.94, P < 0.0001) was observed. The in vitro intestinal digestibility of protein (ivID) quadratically decreased (R2 = 0.99, P < 0.0001) with TP inclusion and the rumen undegradable protein (RUP) showed a quadratic increase (R2 = 0.94, P = 0.0417). Total intestinal protein availability, computed from the RUP and ivID, linearly decreased with TP inclusion level (R2 = 0.45, P = 0.0033).

  13. Effects of degradable protein and non-fibre carbohydrates on microbial growth and fermentation in the rumen simulating fermenter (Rusitec

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    Xiang H. Zhao

    2015-05-01

    Full Text Available A rumen simulation technique (Rusitec apparatus with eight 800 ml fermentation vessels was used to investigate the effects of rumen degradable protein (RDP level and non-fibre carbohydrate (NFC type on ruminal fermentation, microbial growth, and populations of ruminal cellulolytic bacteria. Treatments consisted of two NFC types (starch and pectin supplemented with 0 g/d (low RDP or 1.56 g/d (high RDP sodium caseinate. Apparent disappearance of dry matter and organic matter was greater for pectin than for starch treatment (P<0.01 with low or high RDP. A NFC × RDP interaction was observed for neutral detergent fibre disappearance (P=0.01, which was lower for pectin than for starch only under low RDP conditions. Compared with starch, pectin treatment increased the copy numbers of Ruminococcus albus (P≤0.01 and Ruminococcus flavefaciens (P≤0.09, the molar proportion of acetate (P<0.01, the acetate:propionate ratio (P<0.01, and methane production (P<0.01, but reduced the propionate proportion (P<0.01. Increasing dietary RDP increased the production of total VFA (P=0.01, methane (P<0.01, ammonia N (P<0.01, and microbial N (P<0.01. Significant NFC × RDP interaction and interaction tendency were observed for ammonia N production (P=0.01 and daily N flow of total microorganisms (P=0.07, which did not differ under low RDP conditions, but pectin produced greater microbial N and less ammonia N than starch with increased RDP. Results showed NFC type, RDP level, and their interaction affected ruminal fermentation and microbial growth, and under sufficient ruminal degradable N pectin had greater advantage in microbial N synthesis than starch in vitro.

  14. The effect of starch, inulin, and degradable protein on ruminal fermentation and microbial growth in rumen simulation technique

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    Xiang H. Zhao

    2014-03-01

    Full Text Available A rumen simulation technique apparatus with eight 800 mL fermentation vessels was used to investigate the effects of rumen degradable protein (RDP level and non-fibre carbohydrate (NFC type on ruminal fermentation, microbial growth, and populations of ruminal cellulolytic bacteria. Treatments consisted of two NFC types (starch and inulin supplemented with 0 g/d (low RDP or 1.56 g/d (high RDP sodium caseinate. No significant differences existed among dietary treatments in the apparent disappearance of dietary nutrients except for dietary N, which increased with increased dietary RDP (P<0.001. Compared with starch, inulin treatments reduced the molar proportion of acetate (P<0.001, the acetate:propionate ratio (P<0.001, and methane production (P=0.006, but increased the butyrate proportion (P<0.001. Increased dietary RDP led to increases in production of total volatile fatty acid (P=0.014 and methane (P=0.050, various measures of N (P≤0.046, and 16s rDNA copy numbers of Ruminococcus flavefaciens (P≤0.010. Non-fibre carbohydrate source did not affect daily microbial N flow regardless of dietary RDP, but ammonia N production was lower for inulin than for starch treatments under high RDP conditions (P<0.001. Compared with starch treatments, inulin depressed the copy numbers of Fibrobacter succinogenes in solid fraction (P=0.023 and R. flavefaciens in liquid (P=0.017 and solid fractions (P=0.007, but it increased the carboxymethylcellulase activity in solid fraction (P=0.045. Current results suggest that starch and inulin differ in ruminal volatile fatty acid fermentation but have similar effects on ruminal digestion and microbial synthesis in vitro, although inulin suppressed the growth of partial ruminal cellulolytic bacteria.

  15. Effects of vanillin, quillaja saponin, and essential oils on in vitro fermentation and protein-degrading microorganisms of the rumen.

    Science.gov (United States)

    Patra, Amlan K; Yu, Zhongtang

    2014-01-01

    This study investigated the effects of vanillin on methanogenesis and rumen fermentation, and the responses of ruminal protein-degrading bacteria to vanillin (at concentrations of 0, 0.76 and 1.52 g/L), essential oils (clove oil, 1 g/L; origanum oil, 0.50 g/L, and peppermint oil, 1 g/L), and quillaja saponin (at concentration of 0 and 6 g/L) in vitro. Methane production, degradabilities of feed substrate, and ammonia concentration decreased linearly with increasing doses of vanillin. Concentration of total volatile fatty acids also decreased, whereas proportion of butyrate tended to increase linearly with increasing doses of vanillin. Protozoa population decreased, but abundances of Ruminococcus flavefaciens, Prevotella bryantii, Butyrivibrio fibrisolvens, Prevotella ruminicola, Clostridium aminophilum, and Ruminobacter amylophilus increased with increasing doses of vanillin. Origanum and clove oils resulted in lower ammonia concentrations compared to control and peppermint oil. All the tested essential oils decreased abundances of protozoa, Selenomonas ruminantium, R. amylophilus, P. ruminicola and P. bryantii, with the largest decrease resulted from origanum oil followed by clove oil and peppermint oil. The abundances of Megasphaera elsdenii, C. aminophilum, and Clostridium sticklandii were deceased by origanum oil while that of B. fibrisolvens was lowered by both origanum and clove oils. Saponin decreased ammonia concentration and protozoal population, but increased the abundances of S. ruminantium, R. amylophilus, P. ruminicola, and P. bryantii, though the magnitude was small (less than one log unit). The results suggest that reduction of ammonia production by vanillin and saponin may not be caused by direct inhibition of major known proteolytic bacteria, and essential oils can have different inhibitory effects on different proteolytic bacteria, resulting in varying reduction in ammonia production.

  16. Whole linted cottonseed meal (Gossypium hirsutum L. protein and fiber degradability in the rumen

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    Deborah Clea Ruy

    1996-12-01

    3 x 3 change-over design to evaluate the following treatments: A = 0% WLC; B = 6.6% WLC; and C = 15.0% WLC. Sorghum silage contributed with 70% in all three treatments. DM degradability at 48h incubation time was statistically different (p < 0.05 (A = 54.4%; B = 54.2% and C = 58.7%, as well as PB degradability at 12h (A = 40.3%; B = 47.7% and C = 53.1% and ADF degradability at 48h (A = 40.3%; B = 41.2% and C = 45.6%. Ruminal volume, turn overtime and ruminal pH weren’t affected by the experimental diets. Substitution of WLC for cottonseed meal up to 15% diet increased degradability of DM, CP and ADF of WLC.

  17. Ratio of dietary rumen degradable protein to rumen undegradable protein affects nitrogen partitioning but does not affect the bovine milk proteome produced by mid-lactation Holstein dairy cows.

    Science.gov (United States)

    Tacoma, R; Fields, J; Ebenstein, D B; Lam, Y-W; Greenwood, S L

    2017-09-01

    Little is known about the bovine milk proteome or whether it can be affected by diet. The objective of this study was to determine if the dietary rumen degradable protein (RDP):rumen undegradable protein (RUP) ratio could alter the bovine milk proteome. Six Holstein cows (parity: 2.5 ± 0.8) in mid lactation were blocked by days in milk (80 ± 43 d in milk) and milk yield (57.5 ± 6.0 kg) and randomly assigned to treatment groups. The experiment was conducted as a double-crossover design consisting of three 21-d periods. Within each period, treatment groups received diets with either (1) a high RDP:RUP ratio (RDP treatment: 62.4:37.6% of crude protein) or (2) a low RDP:RUP ratio (RUP treatment: 51.3:48.7% of crude protein). Both diets were isonitrogenous and isoenergetic (crude protein: 18.5%, net energy for lactation: 1.8 Mcal/kg of dry matter). To confirm N and energy status of cows, dry matter intake was determined daily, rumen fluid samples were collected for volatile fatty acid analysis, blood samples were collected for plasma glucose, β-hydroxybutyrate, urea nitrogen, and fatty acid analysis, and total 24-h urine and fecal samples were collected for N analysis. Milk samples were collected to determine the general milk composition and the protein profile. Milk samples collected for high-abundance protein analysis were subjected to HPLC analysis to determine the content of α-casein, β-casein, and κ-casein, as well as α-lactalbumin and β-lactoglobulin. Samples collected for low-abundance protein analysis were fractionated, enriched using ProteoMiner treatment, and separated using sodium dodecyl sulfate-PAGE. After excision and digestion, the peptides were analyzed using liquid chromatography (LC) tandem mass spectrometry (MS/MS). The LC-MS/MS data were analyzed using PROC GLIMMIX of SAS (version 9.4, SAS Institute Inc., Cary, NC) and adjusted using the MULTTEST procedure. All other parameters were analyzed using PROC MIXED of SAS. No treatment differences

  18. The effect of dietary rumen degradable protein content on veal calf ...

    African Journals Online (AJOL)

    The objective of this study was to determine the undegradable dietary protein requirements of veal calves. Two experiments were carried out with Holstein bull calves from 3-10 days of age until slaughter at 20 weeks of age. Both experiments were divided into starter and finishing periods. Calves were offered starter pellets ...

  19. Rumen degradability of some feed legume seeds

    OpenAIRE

    González , Javier; Andrés , Santiago

    2003-01-01

    International audience; The aim of this work was to determine the effective degradability (ED) of CP for different feed legume seeds and the possible relationship with their physical and chemical characteristics. The ED was measured using nylon bags and rumen outflow rate techniques on three rumen cannulated wethers fed at 40 g DM$\\cdot$kg$^{-0.75}$, with a 2:1 (on DM basis) hay to concentrate diet. Nine seed samples of the following legume species were tested: lupin (Lupinus albus L., cultiv...

  20. Effect of supplementation with protein differ for rumen degradability on milk production and nutrients utilization in early lactating Sahiwal cows

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    Talat N. Pasha

    2012-01-01

    Full Text Available Early lactating Sahiwal cows (n=24 of approximately similar yield and lactation were selected and randomly divided into four groups of six cows in each. These groups were fed ad libitum four iso- energetic and iso- proteic diets with different rumen undegradable protein (RUP sources: diet A 30% RUP, diet B 40% RUP, diet C 50% RUP and diet D 60% RUP in a completely randomized design. Among nutrients intake, dry matter (DM and crude protein (CP intake was significantly (P<0.01 different, while neutral detergent fibre (NDF and acid detergent fibre (ADF intakes were similar across four diets. DM, CP and NDF digestibility were also different (P<0.05 except, NDF digestibility. Whole milk yield (kg/d and 4% fat corrected milk (FCM (kg/d, fat (g/d and protein (g/d was found maximum on diet B, followed by diet A. Not significant differences were found in fat, solid not fat (SNF, protein, lactose, salts and total solids percentage across all diet except SNF, lactose and salts percentages which were significantly lower (P<0.05 on diet D. Nitrogen intake, balance and utilization were statistically similar across all diets however, nitrogen excretion in milk (g/d and percentage of intake and urine (percentage of intake were significantly different across diets. Nitrogen intake and output varied (P<0.01 across all diets. Nitrogen balance and its utilization were maximum (P<0.001 on diet B, while other diets showed not significant differences among themselves. Based on presenting findings, it is concluded that feed intake, digestibility and production performance was maximum in early lactating Sahiwal cows when fed 40% rumen undegradable protein in total mixed ration based diet.

  1. Effective rumen degradation of dry matter, crude protein and neutral detergent fibre in forage determined by near infrared reflectance spectroscopy

    DEFF Research Database (Denmark)

    Ohlsson, C; Houmøller, L P; Weisbjerg, Martin Riis

    2007-01-01

    The objective of the present study was to examine if near infrared reflectance spectroscopy (NIRS) could be used to predict degradation parameters and effective degradation from scans of original forage samples. Degradability of dry matter (DM), crude protein (CP) and neutral detergent fibre (NDF......) of 61 samples of perennial ryegrass (Lolium perenne L.) and orchardgrass (Dactylis glomerata L.) was tested by using the in situ technique. The grass samples were harvested at three different stages, early vegetative growth, early reproductive growth and late reproductive growth. Degradability...

  2. Precision-feeding dairy heifers a high rumen-degradable protein diet with different proportions of dietary fiber and forage-to-concentrate ratios.

    Science.gov (United States)

    Lascano, G J; Koch, L E; Heinrichs, A J

    2016-09-01

    The objective of this experiment was to determine the effects of feeding a high-rumen-degradable protein (RDP) diet when dietary fiber content is manipulated within differing forage-to-concentrate ratio (F:C) on nutrient utilization of precision-fed dairy heifers. Six cannulated Holstein heifers (486.98±15.07kg of body weight) were randomly assigned to 2 F:C, low- (45% forage; LF) and high-forage (90% forage; HF) diets and to a fiber proportion sequence [33% grass hay and wheat straw (HS), 67% corn silage (CS; low fiber); 50% HS, 50% CS (medium fiber); and 67% HS, 33% CS (high fiber)] within forage proportion administered according to a split-plot, 3×3 Latin square design (16-d periods). Heifers fed LF had greater apparent total-tract organic matter digestibility coefficients (dC), neutral detergent fiber, and cellulose than those fed LC diets. Substituting CS with HS resulted in a linear reduction in dry matter, organic matter, and cellulose dC. Nitrogen dC was not different between F:C or with increasing proportions of HS in diets, but N retention tended to decrease linearly as HS was increased in the diets. Predicted microbial protein flow to the duodenum decreased linearly with HS addition and protozoa numbers HS interacted linearly, exhibiting a decrease as HS increased for LF, whereas no effects were observed for HF. Blood urea N increased linearly as HS was incorporated. The LF-fed heifers had a greater ruminal volatile fatty acids concentration. We noted a tendency for a greater dry matter, and a significantly higher liquid fraction turnover rate for HF diets. There was a linear numerical increase in the liquid and solid fraction turnover rate as fiber was added to the diets. Rumen fermentation parameters and fractional passages (solid and liquid) rates support the reduction in dC, N retention, and microbial protein synthesis observed as more dietary fiber is added to the rations of dairy heifers precision-fed a constant proportion of rumen-degradable

  3. Protein Secondary Structures (α-helix and β-sheet) at a Cellular Level and Protein Fractions in Relation to Rumen Degradation Behaviours of Protein: A New Approach

    International Nuclear Information System (INIS)

    Yu, P.

    2007-01-01

    Studying the secondary structure of proteins leads to an understanding of the components that make up a whole protein, and such an understanding of the structure of the whole protein is often vital to understanding its digestive behaviour and nutritive value in animals. The main protein secondary structures are the α-helix and β-sheet. The percentage of these two structures in protein secondary structures influences protein nutritive value, quality and digestive behaviour. A high percentage of β-sheet structure may partly cause a low access to gastrointestinal digestive enzymes, which results in a low protein value. The objectives of the present study were to use advanced synchrotron-based Fourier transform IR (S-FTIR) microspectroscopy as a new approach to reveal the molecular chemistry of the protein secondary structures of feed tissues affected by heat-processing within intact tissue at a cellular level, and to quantify protein secondary structures using multicomponent peak modelling Gaussian and Lorentzian methods, in relation to protein digestive behaviours and nutritive value in the rumen, which was determined using the Cornell Net Carbohydrate Protein System. The synchrotron-based molecular chemistry research experiment was performed at the National Synchrotron Light Source at Brookhaven National Laboratory, US Department of Energy. The results showed that, with S-FTIR microspectroscopy, the molecular chemistry, ultrastructural chemical make-up and nutritive characteristics could be revealed at a high ultraspatial resolution (∼10 μm). S-FTIR microspectroscopy revealed that the secondary structure of protein differed between raw and roasted golden flaxseeds in terms of the percentages and ratio of α-helixes and β-sheets in the mid-IR range at the cellular level. By using multicomponent peak modelling, the results show that the roasting reduced (P <0.05) the percentage of α-helixes (from 47.1% to 36.1%: S-FTIR absorption intensity), increased the

  4. Protein Secondary Structures (alpha-helix and beta-sheet) at a Cellular Levle and Protein Fractions in Relation to Rumen Degradation Behaviours of Protein: A New Approach

    Energy Technology Data Exchange (ETDEWEB)

    Yu,P.

    2007-01-01

    Studying the secondary structure of proteins leads to an understanding of the components that make up a whole protein, and such an understanding of the structure of the whole protein is often vital to understanding its digestive behaviour and nutritive value in animals. The main protein secondary structures are the {alpha}-helix and {beta}-sheet. The percentage of these two structures in protein secondary structures influences protein nutritive value, quality and digestive behaviour. A high percentage of {beta}-sheet structure may partly cause a low access to gastrointestinal digestive enzymes, which results in a low protein value. The objectives of the present study were to use advanced synchrotron-based Fourier transform IR (S-FTIR) microspectroscopy as a new approach to reveal the molecular chemistry of the protein secondary structures of feed tissues affected by heat-processing within intact tissue at a cellular level, and to quantify protein secondary structures using multicomponent peak modelling Gaussian and Lorentzian methods, in relation to protein digestive behaviours and nutritive value in the rumen, which was determined using the Cornell Net Carbohydrate Protein System. The synchrotron-based molecular chemistry research experiment was performed at the National Synchrotron Light Source at Brookhaven National Laboratory, US Department of Energy. The results showed that, with S-FTIR microspectroscopy, the molecular chemistry, ultrastructural chemical make-up and nutritive characteristics could be revealed at a high ultraspatial resolution ({approx}10 {mu}m). S-FTIR microspectroscopy revealed that the secondary structure of protein differed between raw and roasted golden flaxseeds in terms of the percentages and ratio of {alpha}-helixes and {beta}-sheets in the mid-IR range at the cellular level. By using multicomponent peak modelling, the results show that the roasting reduced (P <0.05) the percentage of {alpha}-helixes (from 47.1% to 36.1%: S

  5. Reducing crude protein and rumen degradable protein with a constant concentration of rumen undegradable protein in the diet of dairy cows: Production performance, nutrient digestibility, nitrogen efficiency, and blood metabolites.

    Science.gov (United States)

    Bahrami-Yekdangi, M; Ghorbani, G R; Khorvash, M; Khan, M A; Ghaffari, M H

    2016-02-01

    The goals of ruminant protein nutrition are to provide adequate amounts of RDP for optimal ruminal efficiency and to obtain the desired animal productivity with a minimum amount of dietary CP. The aim of the present study was to examine effects of decreasing dietary protein by decreasing RDP with the optimum concentration of RUP on production performance, nutrient digestibility, N retention, rumen fermentation parameters, and blood metabolites in high-producing Holstein cows in early lactation. Nine multiparous lactating cows (second parities, averaging 50 ± 12 d in milk and milk yield of 48 ± 5 kg/d) were used in a triplicate 3 × 3 Latin square design with 3 rations: 1) a total mixed ration (TMR) containing 16.4% CP (10.9% RDP based on DM), 2) a TMR containing 15.6% CP (10% RDP), and 3) a TMR containing 14.8% CP (9.3% RDP). The level of RUP was constant at 5.5% DM across the treatments. All diets were calculated to supply a postruminal lysine to methionine ratio of about 3:1. Dry matter intake, milk yield and composition, 4% fat-corrected milk, and energy-corrected milk were not significantly affected by decreasing dietary CP and RDP levels. Cows fed 16.4% CP diets had greater ( RUP and fecal N excretion (g/d) did not change. Apparent digestibility of nutrients, ruminal pH, and NH-N concentration were not affected with decreasing dietary CP and RDP levels. Apparent N efficiency increased, and RDP N intake and predicted urine N output decreased with decreased concentration of dietary CP and RDP in the diets ( RUP.

  6. Effects of dietary crude protein and rumen-degradable protein concentrations on urea recycling, nitrogen balance, omasal nutrient flow, and milk production in dairy cows.

    Science.gov (United States)

    Mutsvangwa, T; Davies, K L; McKinnon, J J; Christensen, D A

    2016-08-01

    The objective of this study was to determine how interactions between dietary crude protein (CP) and rumen-degradable protein (RDP) concentrations alter urea-nitrogen recycling, nitrogen (N) balance, omasal nutrient flow, and milk production in lactating Holstein cows. Eight multiparous Holstein cows (711±21kg of body weight; 91±17d in milk at the start of the experiment) were used in a replicated 4×4 Latin square design with a 2×2 factorial arrangement of dietary treatments and 29-d experimental periods. Four cows in one Latin square were fitted with ruminal cannulas to allow ruminal and omasal sampling. The dietary treatment factors were CP (14.9 vs. 17.5%; dry matter basis) and RDP (63 vs. 69% of CP) contents. Dietary RDP concentration was manipulated by including unprocessed or micronized canola meal. Diet adaptation (d 1-20) was followed by 8d (d 21-29) of sample and data collection. Continuous intrajugular infusions of [(15)N(15)N]-urea (220mg/d) were conducted for 4d (d 25-29) with concurrent total collections of urine and feces to estimate N balance and whole-body urea kinetics. Proportions of [(15)N(15)N]- and [(14)N(15)N]-urea in urinary urea, and (15)N enrichment in feces were used to calculate urea kinetics. For the low-CP diets, cows fed the high-RDP diet had a greater DM intake compared with those fed the low-RDP diet, but the opposite trend was observed for cows fed the high-CP diets. Dietary treatment had no effect on milk yield. Milk composition and milk component yields were largely unaffected by dietary treatment; however, on the low-CP diets, milk fat yield was greater for cows fed the low-RDP diet compared with those fed the high-RDP diet, but it was unaffected by RDP concentration on the high-CP diets. On the high-CP diets, milk urea nitrogen concentration was greater in cows fed the high-RDP diet compared with those fed the low-RDP diet, but it was unaffected by RDP concentration on the low-CP diets. Ruminal NH3-N concentration tended to

  7. Fractional rate of degradation (kd) of starch in the rumen and its relation to in vivo rumen and total digestibility

    DEFF Research Database (Denmark)

    Hvelplund, Torben; Larsen, Mogens; Lund, Peter

    2009-01-01

    in different ways both chemically and physically. The starch sources were fed in mixed diets together with grass silage and soya bean meal and allocated ad libitum to fistulated dairy cows. The starch content varied between 13 and 35% in ration dry matter for the different starch sources. The design...... was a series of cross-over experiments with two cows and two periods. Ruminal starch pool was estimated from rumen evacuation and starch flow was estimated by duodenal and faeces sampling. Fractional rate of rumen degradation was estimated from the equation [kd = rumen degraded/rumen pool] and rumen and total...

  8. Lowering rumen-degradable protein maintained energy-corrected milk yield and improved nitrogen-use efficiency in multiparous lactating dairy cows exposed to heat stress.

    Science.gov (United States)

    Kaufman, J D; Kassube, K R; Ríus, A G

    2017-10-01

    The objective of this study was to examine the effect of reducing rumen-degradable protein (RDP) and rumen-undegradable protein (RUP) proportions on feed intake, milk production, and N-use efficiency in primiparous and multiparous cows exposed to warm climates. Eighteen primiparous and 30 multiparous mid-lactation Holstein cows were used in a completely randomized design with a 2 × 2 factorial arrangement of treatments. Cows were randomly assigned to 1 of 4 dietary treatments formulated to contain 2 proportions of RDP (10 and 8%) and 2 proportions RUP (8 and 6%) of dry matter (DM) indicated as follows: (1) 10% RDP, 8% RUP; (2) 8% RDP, 8% RUP; (3) 10% RDP, 6% RUP; and (4) 8% RDP, 6% RUP. Protein sources were manipulated to obtain desired RDP and RUP proportions. Diets were isoenergetic and contained 50% forage and 50% concentrate (DM basis). Cows were individually fed the 10% RDP, 8% RUP diet 3 wk before treatment allocation. Cows were exposed to the prevailing Tennessee July and August temperature and humidity in a freestall barn with no supplemental cooling. Main effects and their interaction were tested using the Mixed procedure of SAS (least squares means ± standard error of the mean; SAS Institute Inc., Cary, NC). Observed values of nutrient intake and milk production were used to obtain NRC (2001) model predictions. Cows showed signs of heat stress throughout the study. Reducing from 10 to 8% RDP decreased dry matter intake (DMI; 0.9 kg/d) at 8% RUP, but increased DMI (2.6 kg/d) at 6% RUP in primiparous cows. Reducing from 10 to 8% RDP decreased milk yield (10%) at 8% RUP, but increased yield (14%) at 6% RUP. Treatments did not affect yield of energy-corrected milk. For multiparous cows, treatments did not affect DMI. Reducing from 10 to 8% RDP decreased yield of energy-corrected milk (3.4%) at 8% RUP, but increased yield (8.8%) at 6% RUP. Reducing from 10 to 8% RDP and 8 to 6% RUP both increased N-use efficiency for primiparous and multiparous cows. The NRC

  9. Rumen

    DEFF Research Database (Denmark)

    Mackie, Roderick I.; McSweeney, Christopher S.; Aminov, Rustam

    2013-01-01

    The rumen is a large pregastric fermentation compartment (foregut), which maintains a diverse but concentrated population of anaerobic bacteria, protozoa and fungi that are responsible for a variety of degradative and fermentative reactions. During this process biodegradable organic matter, mainly......). The concept of interspecies hydrogen transfer in which the mutually beneficial unidirectional transfer of hydrogen from a hydrogenproducing to a hydrogenutilising bacteria in a coupled reaction that maintains low partial pressures, which makes the transfer process hermodynamically feasible is important...... in ruminal methanogenesis. Currently, modern ‘Omics’ technologies are being applied to the study of rumen microbial ecology, genomics, metagenomics and metatranscriptomics....

  10. Effects of ratios of non-fibre carbohydrates to rumen degradable ...

    African Journals Online (AJOL)

    To evaluate the effect of different ratios of non-fibre carbohydrates (NFC) to rumen degradable protein (RDP) on lactation responses, digestion and dry matter intake (DMI), nine multiparous mid-lactation Holstein cows, averaging 171 +-17 days in milk and 24.1+-3.3 kg of milk/d were assigned to a 3 x 3 Latin square design.

  11. Effect of rumen-degradable intake protein supplementation on urea kinetics and microbial use of recycled urea in steers consuming low-quality forage.

    Science.gov (United States)

    Wickersham, T A; Titgemeyer, E C; Cochran, R C; Wickersham, E E; Gnad, D P

    2008-11-01

    We evaluated the effect of increasing amounts of rumen-degradable intake protein (DIP) on urea kinetics in steers consuming prairie hay. Ruminally and duodenally fistulated steers (278 kg of BW) were used in a 4 x 4 Latin square and provided ad libitum access to low-quality prairie hay (4.9% CP). The DIP was provided as casein dosed ruminally once daily in amounts of 0, 59, 118, and 177 mg of N/kg of BW daily. Periods were 13 d long, with 7 d for adaptation and 6 d for collection. Steers were in metabolism crates for total collection of urine and feces. Jugular infusion of (15)N(15)N-urea, followed by determination of urinary enrichment of (15)N(15)N-urea and (14)N(15)N-urea was used to determine urea kinetics. Forage and N intake increased (linear, P Urea synthesis was 19.9, 24.8, 42.9, and 50.9 g of urea-N/d for 0, 59, 118, and 177 mg of N/kg of BW daily (linear, P = 0.004). Entry of urea into the gut was 98.9, 98.8, 98.6, and 95.9% of production for 0, 59, 118, and 177 mg of N/kg of BW daily, respectively (quadratic, P = 0.003). The amount of urea-N entering the gastrointestinal tract was greatest for 177 mg of N/kg of BW daily (48.6 g of urea-N/d) and decreased (linear, P = 0.005) to 42.4, 24.5, and 19.8 g of urea-N/d for 118, 59, and 0 mg of N/kg of BW daily. Microbial incorporation of recycled urea-N increased linearly (P = 0.02) from 12.3 g of N/d for 0 mg of N/kg of BW daily to 28.9 g of N/d for 177 mg of N/kg of BW daily. Provision of DIP produced the desired and previously observed increase in forage intake while also increasing N retention. The large percentage of urea synthesis that was recycled to the gut (95.9% even when steers received the greatest amount of DIP) points to the remarkable ability of cattle to conserve N when fed a low-protein diet.

  12. Rumen undegradable protein (RUP) and its intestinal digestibility after steam flaking of cereal grains

    DEFF Research Database (Denmark)

    Chrenková, M; Formelová, Z; Ceresnáková, Z

    2018-01-01

    While it is known that heat treatment of cereal grains generally improves the nutritional value for ruminants, simultaneous information on rumen degradability and intestinal digestibility of the rumen by-pass is scarce, especially for non-starch constituents. The effect of steam flaking at 90°C...... flaking on chemical composition of cereal grains (crude protein, acid detergent fibre, neutral detergent fibre, and starch) were observed. The protein fractions that are relevant to rumen degradability were significantly influenced by the steam flaking: the non-protein nitrogen fraction (A) was reduced (P...... hand, steam flaking markedly increased buffer insoluble but neutral detergent soluble protein fraction (B2) by 15–25% for all three cereal grains, whereas effects on B3 fraction were not significant. Steam flaking was also associated with an increase of the rumen undegradable protein fraction (C...

  13. Yields Components and 48-H Rumen Dry Matter Degradation of ...

    African Journals Online (AJOL)

    Two experiments were conducted to study the influence of harvesting date on three sweet potato varieties (TIS-87/0087, TIS-8164 and TIS-2532.OP.1.13). Fodder yields and leaf-to-stem ratio decreased (P < 0.05), while harvest index and 48-h rumen DM degradation increased with maturity from 12 to 20 weeks after ...

  14. In situ rumen degradability characteristics of rice straw, soybean ...

    African Journals Online (AJOL)

    In situ rumen degradability characteristics of rice straw, soybean curd residue and peppermint (Mentha piperita) in Hanwoo steer (Bos Taurus coreanae). Byong Tae Jeon, KyoungHoon Kim, Sung Jin Kim, Na Yeon Kim, Jae Hyun Park, Dong Hyun Kim, Mi Rae Oh, Sang Ho Moon ...

  15. Degradation of spent craft brewer's yeast by caprine rumen hyper ammonia-producing bacteria.

    Science.gov (United States)

    Harlow, B E; Bryant, R W; Cohen, S D; O'Connell, S P; Flythe, M D

    2016-10-01

    Spent yeast from craft beers often includes more hops (Humulus lupulus L.) secondary metabolites than traditional recipes. These compounds include α- and β- acids, which are antimicrobial to the rumen hyper ammonia-producing bacteria (HAB) that are major contributors to amino acid degradation. The objective was to determine if the hops acids in spent craft brewer's yeast (CY; ~ 3·5 mg g(-1) hops acids) would protect it from degradation by caprine rumen bacteria and HAB when compared to a baker's yeast (BY; no hops acids). Cell suspensions were prepared by harvesting rumen fluid from fistulated goats, straining and differential centrifugation. The cells were re-suspended in media with BY or CY. After 24 h (39°C), HAB were enumerated and ammonia was measured. Fewer HAB and less ammonia was produced from CY than from BY. Pure culture experiments were conducted with Peptostreptococcus anaerobiusBG1 (caprine HAB). Ammonia production by BG1 from BY was greater than from CY. Ammonia production was greater when exogenous amino acids were included, but similar inhibition was observed in CY treatments. These results indicate that rumen micro-organisms deaminated the amino acids in CY to a lesser degree than BY. Spent brewer's yeast has long been included in ruminant diets as a protein supplement. However, modern craft beers often include more hops (Humulus lupulus L.) than traditional recipes. These compounds include α- and β- acids, which are antimicrobial to the rumen hyper ammonia-producing bacteria (HAB) that are major contributors to amino acid degradation. This study demonstrated that hops acids in spent craft brewer's yeast protected protein from destruction by HABin vitro. These results suggest that the spent yeast from craft breweries, a source of beneficial hops secondary metabolites, could have value as rumen-protected protein. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

  16. Determination of in vitro isoflavone degradation in rumen fluid.

    Science.gov (United States)

    Trnková, Andrea; Šancová, Kateřina; Zapletalová, Martina; Kašparovská, Jitka; Dadáková, Kateřina; Křížová, Ludmila; Lochman, Jan; Hadrová, Sylvie; Ihnatová, Ivana; Kašparovský, Tomáš

    2018-06-01

    The aim of this study was to determine the degradation of dietary isoflavones in rumen fluid under 2 feeding regimens. The experiments were performed in vitro using a rumen fluid buffer system. The rumen fluid was taken from cows fed either a hay diet or a concentrate-rich diet (the diet consisted of 34.6% maize silage, 17.6% haylage, 12.8% alfalfa hay, and 35.0% supplemental mixture on a dry matter basis). As a source of isoflavones, 40% soybean extract (Biomedica, Prague, Czech Republic) at levels of 5, 25, 50, and 75 mg per 40 mL of rumen fluid was used. Samples of soybean extract were incubated in triplicate at 39°C for 0, 3.0, 6.0, 12.0, and 24.0 h in incubation solution. The metabolism of daidzein and genistein was faster under concentrate-rich diet conditions. In general, production of equol started after 3 to 6 h of incubation and reached the highest rate after approximately 12 h of incubation regardless of the type of diet or concentration of extract. In most of the experiments, production of equol continued after 24 h of incubation. Generally, equol production was greater under the hay diet conditions. Furthermore, experiments with higher amounts of added soybean extract revealed possible inhibitory effects of high levels of isoflavones on the rumen microflora. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  17. The Effect of Selected Biostimulating Substance on the Degradation in the Rumen

    Directory of Open Access Journals (Sweden)

    Eva Petrášková

    2011-05-01

    Full Text Available The cows with the canulla were divided into two groups - experimental and control. One cow was added to experimental product Biopolym FZT, second Biopolym granules. The samples of rumen fluid and feces were analyzed. The growth of microorganisms in the rumen fluid of experimental animals means the possibility of a positive impact of Biopolym on the degradation of feeding the rumen.

  18. Degradation of Dehydrodivanillin by Anaerobic Bacteria from Cow Rumen Fluid

    OpenAIRE

    Chen, Wei; Ohmiya, Kunio; Shimizu, Shoichi; Kawakami, Hidekuni

    1985-01-01

    Dehydrodivanillin (DDV; 0.15 g/liter) was biodegradable at 37°C under strictly anaerobic conditions by microflora from cow rumen fluid to the extent of 25% within 2 days in a yeast extract medium. The anaerobes were acclimated on DDV for 2 weeks, leading to DDV-degrading microflora with rates of degradation eight times higher than those initially. Dehydrodivanillic acid and vanillic acid were detected in an ethylacetate extract of a DDV-enriched culture broth by thin-layer, gas, and high-perf...

  19. [Screening of harmine tolerance/degrading bacteria from camel rumen].

    Science.gov (United States)

    An, Dengdi; Zhu, Yanlei; Tang, Jing; Ye, Yongxia; Zeng, Xianchun

    2010-08-01

    Peganum harmala, a famous traditional Chinese drug, contains a variety of alkaloids and toxic for many animals. Camels mainly live in desert or semi-desert areas, with the robust gastrointestine system in digesting various feed including toxic plants without disease symptoms. Camel rumen content was used as the inoculant to inoculate medium M98-5 which contains 100 mg x L(-1) harmin and cultivated for 5 days. Upto 5 subculturings, strains that could degrading or tolerant harmine were isolated. Their conversion activity was determined by thin-layer chromatography. The taxonomic position of the strains were identified based on 16S rRNA sequences analysis. 15 out of the 29 isolates have harmine degrading activity. Most of the isolates are identified as the members of the Genera Lactobacillus (16 strains, 55%), Shigella (7 strains, 24%) and Bacillus (4 strains, 13.8%). Only one strain belong to genus Enterococcus and one belong to genus Megasphaera. The results indicated that the harmine tolerance/degrading communities of camel rumen are limited and only Lactobacillus have harmine-degrading activity.

  20. Chemical composition, rumen degradability, protein utilization and lactation response to selected tree leaves as substitute of cottonseed cake in the diet of dairy goats

    NARCIS (Netherlands)

    Khan, N.A.; Habib, G.; Ullah, G.

    2009-01-01

    The aim of this study was to investigate the potential of leaves from Grewia oppositifolia (G. oppositifolia) and Ziziphus mauritiana (Z. mauritiana) as a crude protein (CP) supplements to low quality diets of goats in Pakistan. Chemical composition and CP degradability of the tree leaves were

  1. Effects of sequence of nylon bags rumen incubation on kinetics of degradation in some commonly used feedstuffs in dairy rations

    Institute of Scientific and Technical Information of China (English)

    DONG Shuang-zhao; Arash Azarfar; ZOU Yang; LI Sheng-li; WANG Ya-jing; CAO Zhi-jun

    2017-01-01

    Nowadays, most available information on the degradative behaviour of feeds in ruminants is based onin situ incubation in the rumen, and it is adopted by many feed evaluation systems currently in use for ruminants. However, the outcome of this technique might be affected by many factors such as sequence of nylon bags incubation in the rumen. The objective of current study was to investigate effects of sequence of nylon bag incubation on degradative behavior of dry matter (DM), crude protein (CP), neutral detergent ifber (NDF) and acid detergent ifber (ADF) in some feed ingredients commonly used in dairy rations, including alfalfa haylage, corn silage, corn grain and soybean meal. Four multiparous Holstein lactating cows iftted with permanent ruminal cannulas were used. The nylon bags containing feed samples either were placed in the rumen at once and removed at designated time intervals (all in-gradually out method; AG) or were placed in the rumen at designated time points and retrieved at once (gradually in-all out method; GA). Fractional rate of degradation of potentially degradable fraction, lag time and effective rumen degradability (ED) of DM and CP were signiifcantly higher in the AG compared to the GA method (P namely corn grain and soybean meal. This experiment is the ifrst time to investigate effects of two methods under the same experiment conditions, providing basic data for the determination of ED.

  2. degradable protein sources on performance of high-producing dairy ...

    African Journals Online (AJOL)

    with high-quality, low·degradable protein sources prOViding47% UDP is advocated for ... saliva and through the rumen wall (Waldo, 1968). Based on this type of ... of the feed industry, but is based on very little solid evidence. (Huber, 1984). Chalupa ...... of rumen fermentation in relation to ammonia concentration. Br. J. Nutr.

  3. Degradation of Crude Protein in Groundnut Cake, Guinea Grass ...

    African Journals Online (AJOL)

    Three West African dwarf rams fitted with rumen cannula, were used in a completely randomized design for degradation of crude protein (CP) of groundnut cake (GNC), Panicum maximum, rumen epithelial scraping (RES), and diets containing increasing levels of RES. Concentrate diets were formulated such that 0% (A), ...

  4. Shorten fungal treatment of lignocellulosic waste with additives to improve rumen degradability

    NARCIS (Netherlands)

    Kuijk, van S.J.A.; Sonnenberg, A.S.M.; Baars, J.J.P.; Hendriks, W.H.; Cone, J.W.

    2014-01-01

    Selective lignin degrading fungi can be used as pre-treatment to make cellulose in plant cell walls accessible for rumen microbes. According to previous studies, Ceriporiopsis subvermispora and Lentinula edodes can increase the in vitro rumen degradability of lignocellulosic biomass in 7 to 8 weeks.

  5. The effect of the supply of rumen degradable protein and metabolisable protein on negative energy balance and fertility in dairy cows

    NARCIS (Netherlands)

    Tamminga, S.

    2006-01-01

    Reproduction in dairy cattle is negatively affected by a negative energy balance (NEB), a combination of the deposition or mobilisation of fat and protein. The mode of action of NEB on fertility is not always clear, but the severity, length, and probably also the nature of the NEB may be involved.

  6. Digestion of crude protein and organic matter of leaves by rumen microbes in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Ciszuk, A.; Murphy, M.

    1982-01-01

    22 leaf specimens, of which 6 were from an energy-woods project, were studied by incubation in vitro with rumen microbes or pepsin-hydrochloric acid. Several were also examined in situ using the nylon-bag technique. Many leaves, despite their low fiber and high crude protein content, gave low values for organic matter digestibility. The crude protein degradation by rumen microbes or pepsin-hydrochloric acid was low, on average, compared with hay. There was a wide variation among leaf specimens. Variation was also found as regards ammonia production in short-term (4 hours) incubation. No close correlation was found between crude protein content and crude protein degradation, or between the estimates of ruminal degradation and of pepsin-hydrochloric acid digestibility. This suggest that there are leaves that gives ruminants substantial amounts of digestible protein yet escape ruminal fermentation. (Refs. 12).

  7. Effects of inclusion levels of banana (Musa spp.) peelings on feed degradability and rumen environment of cattle fed basal elephant grass.

    Science.gov (United States)

    Nambi-Kasozi, Justine; Sabiiti, Elly Nyambobo; Bareeba, Felix Budara; Sporndly, Eva; Kabi, Fred

    2016-04-01

    The effect of feeding varying banana peeling (BP) levels on rumen environment and feed degradation characteristics was evaluated using three rumen fistulated steers in four treatments. The steers were fed BP at 0, 20, 40, and 60% levels of the daily ration with basal elephant grass (EG) to constitute four diets. Maize bran, cotton seed cake, and Gliricidia sepium were offered to make the diets iso-nitrogenous. The nylon bag technique was used to measure BP and EG dry matter (DM), crude protein (CP), and neutral detergent fiber (NDF) degradabilities at 0, 6, 12, 24, 48, 72, 96, and 120 h. Rumen fluid samples were collected to determine pH and volatile fatty acid (VFA) concentrations. Effective DM, CP, and NDF degradabilities of BP ranged between 574 and 807, 629-802, and 527-689 g/kg, respectively, being lower at higher BP levels. Elephant grass degradability behaved similarly with relatively high effective CP degradability (548-569 g/kg) but low effective DM and NDF degradability (381-403 and 336-373 g/kg, respectively). Rumen pH and VFA reduced with increasing BP in the diets. Rumen pH dropped to 5.8 and 5.9 at the 40 and 60% BP feeding levels, respectively. Banana peelings were better degraded than EG but higher BP levels negatively affected feed degradability and rumen environment.

  8. Potential of Selected Rumen Bacteria for Cellulose and Hemicellulose Degradation

    Directory of Open Access Journals (Sweden)

    Maša Zorec

    2014-01-01

    Full Text Available Herbivorous animals harbour potent cellulolytic and hemicellulolytic microorganisms that supply the host with nutrients acquired from degradation of ingested plant material. In addition to protozoa and fungi, rumen bacteria contribute a considerable part in the breakdown of recalcitrant (hemicellulosic biomass. The present review is focused on the enzymatic systems of three representative fibrolytic rumen bacteria, namely Ruminococcus flavefaciens, Prevotella bryantii and Pseudobutyrivibrio xylanivorans. R. flavefaciens is known for one of the most elaborated cellulosome architectures and might represent a promising candidate for the construction of designer cellulosomes. On the other hand, Prevotella bryantii and Pseudobutyrivibrio xylanivorans produce multiple free, but highly efficient xylanases. In addition, P. xylanivorans was also shown to have some probiotic traits, which makes it a promising candidate not only for biogas production, but also as an animal feed supplement. Genomic and proteomic analyses of cellulolytic and hemicellulolytic bacterial species aim to identify novel enzymes, which can then be cloned and expressed in adequate hosts to construct highly active recombinant hydrolytic microorganisms applicable for different biotechnological tasks.

  9. Effects of sheep breed and soybean meal supplementation on rumen environment and degradation kinetics

    NARCIS (Netherlands)

    Lourenco, A.; Cone, J.W.; Fontes, P.; Dias-Da-Silva, A.

    2013-01-01

    The aim of this study was to evaluate if the in vivo digestibility and intake differences, observed in previous studies, between Ile-de-France (IF) and Churra-da-Terra-Quente (CTQ) sheep breeds, were due to rumen environment and degradability differences. The intake, digestibility, rumen environment

  10. Degradation of lucerne stem cell walls by five rumen bacterial species

    NARCIS (Netherlands)

    Jung, H.G.; Engels, F.M.; Weimer, P.J.

    2004-01-01

    The rumen bacterial strains Butyrivibrio fibrisolvens H17c, Fibrobacter succinogenes S85, Lachnospira multiparus 40, Ruminococcus albus 7 and R. flavefaciens FD-1 were compared individually and as a five-species mixture with a rumen inoculum for their ability to degrade lucerne (Medicago sativa L.)

  11. Effect of different levels of rapidly degradable carbohydrates calculated by a simple rumen model on performance of lactating dairy cows.

    Science.gov (United States)

    Doorenbos, J; Martín-Tereso, J; Dijkstra, J; van Laar, H

    2017-07-01

    Aggregating rumen degradation characteristics of different carbohydrate components into the term modeled rapidly degradable carbohydrates (mRDC) can simplify diet formulation by accounting for differences in rate and extent of carbohydrate degradation within and between feedstuffs. This study sought to evaluate responses of lactating dairy cows to diets formulated with increasing levels of mRDC, keeping the supply of other nutrients as constant as possible. The mRDC content of feedstuffs was calculated based on a simple rumen model including soluble, washable, and nonwashable but potentially degradable fractions, as well as the fractional degradation and passage rates, of sugar, starch, neutral detergent fiber, and other carbohydrates. The mRDC term effectively represents the total amount of carbohydrates degraded in the rumen within 2 h after ingestion. Fifty-two lactating Holstein cows (of which 4 were rumen fistulated) were assigned to 4 treatments in a 4 × 4 Latin square design. Treatments were fed as a total mixed ration consisting of 25.4% corn silage, 23.1% grass silage, 11.6% grass hay, and 39.9% concentrate on a dry matter basis. Differences in mRDC were created by exchanging nonforage neutral detergent fiber-rich ingredients (mainly sugar beet pulp) with starch-rich ingredients (mainly wheat) and by exchanging corn (slowly degradable starch) with wheat (rapidly degradable starch) in the concentrate, resulting in 4 treatments that varied in dietary mRDC level of 167, 181, 194, or 208 g/kg of dry matter. Level of mRDC did not affect dry matter intake. Fat- and protein-corrected milk production and milk fat and lactose yield were greatest at 181 mRDC and decreased with further increases in mRDC. Milk protein yield and concentration increased with increasing mRDC level. Mean rumen pH and diurnal variation in ruminal pH did not differ between treatments. Total daily meal time and number of visits per meal were smaller at 181 and 194 mRDC. Despite milk

  12. Fourier transform infrared microspectroscopic analysis of the effects of cereal type and variety within a type of grain on structural makeup in relation to rumen degradation kinetics.

    Science.gov (United States)

    Walker, Amanda M; Yu, Peiqiang; Christensen, Colleen R; Christensen, David A; McKinnon, John J

    2009-08-12

    The objectives of this study were to use Fourier transform infrared microspectroscopy (FTIRM) to determine structural makeup (features) of cereal grain endosperm tissue and to reveal and identify differences in protein and carbohydrate structural makeup between different cereal types (corn vs barley) and between different varieties within a grain (barley CDC Bold, CDC Dolly, Harrington, and Valier). Another objective was to investigate how these structural features relate to rumen degradation kinetics. The items assessed included (1) structural differences in protein amide I to nonstructural carbohydrate (NSC, starch) intensity and ratio within cellular dimensions; (2) molecular structural differences in the secondary structure profile of protein, alpha-helix, beta-sheet, and their ratio; (3) structural differences in NSC to amide I ratio profile. From the results, it was observed that (1) comparison between grain types [corn (cv. Pioneer 39P78) vs barley (cv. Harrington)] showed significant differences in structural makeup in terms of NSC, amide I to NSC ratio, and rumen degradation kinetics (degradation ratio, effective degradability of dry matter, protein and NSC) (P makeup in terms of amide I, NSC, amide I to NSC ratio, alpha-helix and beta-sheet protein structures, and rumen degradation kinetics (effective degradability of dry matter, protein, and NSC) (P makeup differences between cereal types and between different varieties within a type of grain could be revealed. These structural makeup differences were related to the rate and extent of rumen degradation.

  13. Isolation and some characteristics of anaerobic oxalate-degrading bacteria from the rumen.

    OpenAIRE

    Dawson, K A; Allison, M J; Hartman, P A

    1980-01-01

    Obligately anaerobic oxalate-degrading bacteria were isolated from an enriched population of rumen bacteria in an oxalate-containing medium that had been depleted of other readily metabolized substrates. These organisms, which are the first reported anaerobic oxalate degraders isolated from the rumen, were gram negative, nonmotile rods. They grew in a medium containing sodium oxalate, yeast extract, cysteine, and minerals. The only substrate that supported growth was oxalate. Growth was direc...

  14. Investigation of the rumen microbial community responsible for degradation of a putative toxin in Acacia angustissima

    International Nuclear Information System (INIS)

    Collins, E.M.C.; Blackall, L.L.; Mcsweeney, C.S.; Krause, D.O.

    2005-01-01

    Acacia angustissima has been proposed as a protein supplement in countries where availability of high quality fodder for grazing animals is a problem due to extreme, dry climates. While A. angustissima thrives in harsh environments and provides valuable nutrients required by ruminants, it has also been found to contain anti-nutritive factors that currently preclude its widespread application. A number of non-protein amino acids have been identified in the leaves of A. angustissima and in the past these have been linked to toxicity in ruminants. The non-protein amino acid 4-n-acetyl-2,4-diaminobutyric acid (ADAB) had been determined to be the major non-protein amino acid in the leaves of A. angustissima. Thus, in this study, the aim was to identify microorganisms from the rumen environment capable of degrading ADAB. Using an ADAB-containing plant extract, a mixed enrichment culture was obtained that exhibited substantial ADAB-degrading ability. Attempts to isolate an ADAB-degrading micro-organism were carried out, but no isolates were able to degrade ADAB in pure culture. The mixed microbial community of the ADAB-degrading enrichment culture was further examined through the use of pure-culture-independent techniques. Fluorescence in situ hybridization (FISH) was employed to investigate the diversity within this sample. In addition two bacterial 16S rDNA clone libraries were constructed in an attempt to further elucidate the members of the microbial population. The clone libraries were constructed from serial dilutions of the enrichment culture, a 10 -5 dilution where complete degradation of ADAB occurred, and a 10 -7 dilution where ADAB degradation did not occur. Through the comparison of these two libraries it was hypothesized that clones belonging to the Firmicutes phylum were involved in ADAB degradation. A FISH probe, ADAB1268, was then designed to target these clones and was applied to the enrichment cultures to investigate their relative abundance within the

  15. Degradabilidade ruminal e digestibilidade intestinal da proteína de capim-elefante com três idades de corte Rumen degradability and intestinal digestibility of protein of elephant-grass at three cutting ages

    Directory of Open Access Journals (Sweden)

    J.P.G. Soares

    2009-04-01

    Full Text Available Determinaram-se a degradabilidade potencial (DP e a digestibilidade intestinal da proteína não degradada no rúmen (DIPNDR do capim-elefante em diferentes idades de rebrote (30, 45 e 60 dias e comparou-se a técnica do saco de náilon móvel (in situ com o método de três estádios (in vitro. Para tanto, utilizaram-se seis novilhos mestiços canulados no rúmen e duodeno alimentados exclusivamente com capim-elefante picado. O ensaio de degradabilidade foi realizado com amostras do capim incubadas no rúmen por 3, 6, 9, 12, 24, 48, 72, 96 e 120h. A digestibilidade intestinal foi determinada utilizando-se os resíduos de incubação por 24 horas. Na técnica in situ os resíduos em sacos de náilon foram colocados no duodeno e recuperados nas fezes. No método in vitro, os resíduos foram submetidos à digestão com HCl-pepsina-pancreatina. Em amostras de capim com idades de 30, 45 e 60 dias foram observados valores de DP da proteína de 87,5; 87,8 e 83,8%, respectivamente. A DIPNDR variou com a idade do capim e foi semelhante entre os métodos in situ e in vitro somente para o capim com 60 dias. O método in situ apresentou estimativa de digestibilidade intestinal mais coerente com as mudanças na composição química do capim-elefante decorrentes do envelhecimento.The potential degradability (PD and intestinal digestibility of ruminal escape protein (IDREP of elephant-grass at 30, 45, and 60 days of regrowth were determined and the mobile bag technique (in situ was compared to the three-stage method (in vitro. Thus, six cross-bred steers with rumen and duodenum canulas were used and fed exclusively with chopped elephant grass. The degradability trial was carried out with grass samples incubated in rumen by 3, 6, 9, 12, 24, 48, 72, 96, and 120 h. The intestinal digestibility was determined using 24-h ruminal incubation residue. In the in situ technique, residues in nylon bags were placed in duodenum and recovered in feces. In the in vitro

  16. Ammonia treatment of wheat straw. 2. Efficiency of microbial protein synthesis, rumen microbial protein pool size and turnover, and small intestinal protein digestion in sheep.

    NARCIS (Netherlands)

    Oosting, S.J.; Viets, T.C.; Lammers-Wienhoven, S.C.W.; Bruchem, van J.

    1993-01-01

    Ammonia-treated wheat straw (AWS) was compared with untreated wheat straw (UWS) and untreated wheat straw supplemented with urea (SWS) in an experiment with 6 wether sheep. Microbial protein synthesis increased after ammonia treatment due to the higher intake of rumen degradable organic matter (OM).

  17. Chemical Composition and Rumen Degradation Characteristics of Different Chickpea (Cicer Arietinum L. Lines Straw

    Directory of Open Access Journals (Sweden)

    Numan Kılıçalp

    2017-06-01

    Full Text Available This study aimed to identfy chemical composition, ruminal degradation characeristics and metabolizable energy (ME content of five different chickpea line and a check cultivar’s straw using nylon bag technique. Feed samples were incubated as three replicates of each fistulated Holstein heifer for 0, 8, 12, 24, 36, 48, 72 and 96 h. Degradation characteristics of dry matter (DM and neutral detergent fiber (NDF in rumen were determined by using this mathematical expression D=a+b(1-e-ct. Crude protein (CP, acid detergent fiber (ADF, neutral detergent fiber (NDF, and ash contents of straw were ranged from 5.61 to 7.42%, 51.33 to 56.0%, 63.67 to 67.0%, and 8.0 to 9.0% respectively. Besides Rapidly soluble fraction (a, potantial degradability (a+b and effective dry matter degradability (EDDM were ranged from 17.86 to 21.41, 54.40 to 59.43, 49.65 to 54.91% respectively. Estimated ME of chickpea entries straw were ranged from 5.96 to 7.37 MJ/kg. Metabolizable energy content of control chickpea cultivar was significantly higher than the other chickpea straw of lines. The research values of ME revealed that significant differences were determined among the lines in terms of energy content. In addition to, a strong relationship between straw NDF level and ME content were determined.

  18. Rumen microbial protein synthesis and nitrogen efficiency as affected by tanniferous and non-tanniferous forage legumes incubated individually or together in Rumen Simulation Technique.

    Science.gov (United States)

    Grosse Brinkhaus, Anja; Bee, Giuseppe; Schwarm, Angela; Kreuzer, Michael; Dohme-Meier, Frigga; Zeitz, Johanna O

    2018-03-01

    A limited availability of microbial protein can impair productivity in ruminants. Ruminal nitrogen efficiency might be optimised by combining high-quality forage legumes such as red clover (RC), which has unfavourably high ruminal protein degradability, with tanniferous legumes like sainfoin (SF) and birdsfoot trefoil (BT). Silages from SF and from BT cultivars [Bull (BB) and Polom (BP)] were incubated singly or in combination with RC using the Rumen Simulation Technique (n = 6). The tanniferous legumes, when compared to RC, changed the total short-chain fatty acid profile by increasing propionate proportions at the expense of butyrate. Silage from SF contained the most condensed tannins (CTs) (136 g CT kg -1 dry matter) and clearly differed in various traits from the BT and RC silages. The apparent nutrient degradability (small with SF), microbial protein synthesis, and calculated content of potentially utilisable crude protein (large with SF) indicated that SF had the greatest efficiency in ruminal protein synthesis. The effects of combining SF with RC were mostly linear. The potential of sainfoin to improve protein supply, demonstrated either individually or in combination with a high-performance forage legume, indicates its potential usefulness in complementing protein-deficient ruminant diets and high-quality forages rich in rumen-degradable protein. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  19. Isolation and Identification of Sodium Fluoroacetate Degrading Bacteria from Caprine Rumen in Brazil

    Directory of Open Access Journals (Sweden)

    Expedito K. A. Camboim

    2012-01-01

    Full Text Available The objective of this paper was to report the isolation of two fluoroacetate degrading bacteria from the rumen of goats. The animals were adult goats, males, crossbred, with rumen fistula, fed with hay, and native pasture. The rumen fluid was obtained through the rumen fistula and immediately was inoculated 100 μL in mineral medium added with 20 mmol L−1 sodium fluoroacetate (SF, incubated at 39°C in an orbital shaker. Pseudomonas fluorescens (strain DSM 8341 was used as positive control for fluoroacetate dehalogenase activity. Two isolates were identified by 16S rRNA gene sequencing as Pigmentiphaga kullae (ECPB08 and Ancylobacter dichloromethanicus (ECPB09. These bacteria degraded sodium fluoroacetate, releasing 20 mmol L−1 of fluoride ion after 32 hours of incubation in Brunner medium containing 20 mmol L−1 of SF. There are no previous reports of fluoroacetate dehalogenase activity for P. kullae and A. dichloromethanicus. Control measures to prevent plant intoxication, including use of fences, herbicides, or other methods of eliminating poisonous plants, have been unsuccessful to avoid poisoning by fluoroacetate containing plants in Brazil. In this way, P. kullae and A. dichloromethanicus may be used to colonize the rumen of susceptible animals to avoid intoxication by fluoroacetate containing plants.

  20. Fractional rate of degradation (kd) of starch in the rumen and its ...

    African Journals Online (AJOL)

    Fractional rate of degradation (kd) of fermentable nutrients in the rumen is an important parameter in modern feed evaluation systems based on mechanistic models. Estimates of kd for starch was obtained on 19 starch sources originating from barley, wheat, oat, maize and peas and treated in different ways both chemically ...

  1. Variation between individual cows in in situ rumen degradation characteristics of maize and grass silages

    NARCIS (Netherlands)

    Ali, M.; Cone, J. W.; van Duinkerken, G.; Klop, A.; Blok, M. C.; Bruinenberg, M.; Khan, N. A.; Hendriks, W. H.

    2016-01-01

    Different numbers of animals have been used in different studies to cover the variation between individual animals in in situ rumen degradation characteristics of maize and grass silages. The objective of this study was to determine whether three cows are sufficient or not to cover the variation

  2. The amino acid composition of rumen-undegradable protein: a comparison between forages.

    Science.gov (United States)

    Edmunds, B; Südekum, K-H; Bennett, R; Schröder, A; Spiekers, H; Schwarz, F J

    2013-07-01

    The objective of this study was to improve knowledge regarding the amino acid profile of the insoluble portion of ingested forage escaping rumen degradation. Six forage categories were analyzed. Categories varied in botanical composition and each contained 2 samples. Samples within categories were derived from the same parent material but differed in harvest, maturity, or conservation type. The rumen-undegradable protein of all forages was measured by incubation for 16h in the rumen of 3 nonlactating cows. All residues were corrected for microbial colonization. The AA profile of the residue was different to the original profile. Degradation trends of individual AA, in terms of increase or decrease relative to the original concentration, were similar between all forages. The AA profiles of forage residues, both within and between categories, were more similar to each other than to their respective original profile. This information may aid in improving the accuracy of estimating postruminal AA supply from forages while decreasing the number of samples required to be analyzed. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  3. Estimation of the True Digestibility of Rumen Undegraded Dietary Protein in the Small Intestine of Ruminants by the Mobile Bag Technique

    DEFF Research Database (Denmark)

    Hvelplund, Torben; Weisbjerg, Martin Riis; Andersen, L. S.

    1992-01-01

    Dietary protein degraded to various extents by varying the time of rumen incubation was prepared from eight concentrates and four roughages. Intestinal digestibility was obtained using the mobile bag technique on intact protein and on the samples of undegraded dietary protein from each feed. The ...

  4. Anaerobic degradation of veratrylglycerol-beta-guaiacyl ether and guaiacoxyacetic acid by mixed rumen bacteria.

    OpenAIRE

    Chen, W; Supanwong, K; Ohmiya, K; Shimizu, S; Kawakami, H

    1985-01-01

    Veratrylglycerol-beta-guaiacyl ether (0.2 g/liter), a lignin model compound, was found to be degraded by mixed rumen bacteria in a yeast extract medium under strictly anaerobic conditions to the extent of 19% within 24 h. Guaiacoxyacetic acid, 2-(o-methoxyphenoxy)ethanol, vanillic acid, and vanillin were detected as degradation products of veratrylglycerol-beta-guaiacyl ether by thin-layer chromatography, gas chromatography, and gas chromatography-mass spectrometry. Guaiacoxyacetic acid (0.25...

  5. Effects of dietary supplementation of rumen-protected folic acid on rumen fermentation, degradability and excretion of urinary purine derivatives in growing steers.

    Science.gov (United States)

    Wang, Cong; Liu, Qiang; Guo, Gang; Huo, WenJie; Ma, Le; Zhang, YanLi; Pei, CaiXia; Zhang, ShuanLin; Wang, Hao

    2016-12-01

    The present experiment was undertaken to determine the effects of dietary addition of rumen-protected folic acid (RPFA) on ruminal fermentation, nutrient degradability, enzyme activity and the relative quantity of ruminal cellulolytic bacteria in growing beef steers. Eight rumen-cannulated Jinnan beef steers averaging 2.5 years of age and 419 ± 1.9 kg body weight were used in a replicated 4 × 4 Latin square design. The four treatments comprised supplementation levels of 0 (Control), 70, 140 and 210 mg RPFA/kg dietary dry matter (DM). On DM basis, the ration consisted of 50% corn silage, 47% concentrate and 3% soybean oil. The DM intake (averaged 8.5 kg/d) was restricted to 95% of ad libitum intake. The intake of DM, crude protein (CP) and net energy for growth was not affected by treatments. In contrast, increasing RPFA supplementation increased average daily gain and the concentration of total volatile fatty acid and reduced ruminal pH linearly. Furthermore, increasing RPFA supplementation enhanced the acetate to propionate ratio and reduced the ruminal ammonia N content linearly. The ruminal effective degradability of neutral detergent fibre from corn silage and CP from concentrate improved linearly and was highest for the highest supplementation levels. The activities of cellobiase, xylanase, pectinase and α-amylase linearly increased, but carboxymethyl-cellulase and protease were not affected by the addition of RPFA. The relative quantities of Butyrivibrio fibrisolvens, Ruminococcus albus, Ruminococcus flavefaciens and Fibrobacter succinogenes increased linearly. With increasing RPFA supplementation levels, the excretion of urinary purine derivatives was also increased linearly. The present results indicated that the supplementation of RPFA improved ruminal fermentation, nutrient degradability, activities of microbial enzymes and the relative quantity of the ruminal cellulolytic bacteria in a dose-dependent manner. According to the conditions of this

  6. Degradation of spent craft brewer’s yeast by caprine rumen hyper ammonia-producing bacteria

    Science.gov (United States)

    Spent brewer’s yeast has long been included in ruminant diets as a protein supplement. However, modern craft beers often include more hops (Humulus lupulus L.) compounds than traditional recipes. These compounds include alpha and beta-acids, which are antimicrobial to the rumen hyper ammonia-produci...

  7. Caracterização, fracionamento protéico, degradabilidade ruminal e digestibilidade in vitro da matéria seca e proteína bruta do resíduo de cervejaria úmido e fermentado = Characterization, protein fractioning, dry matter and crude protein rumen degradability and in vitro digestibility of wet and fermented brewer’s grain

    Directory of Open Access Journals (Sweden)

    Luiz Juliano Valério Geron

    2007-07-01

    Full Text Available Foram avaliadas as frações da proteína e dos carboidratos, a degradabilidade ruminal efetiva (DE da matéria seca (MS e proteína bruta (PB, a digestibilidade ruminal in vitro (DRIV da MS e PB, a digestibilidade intestinal in vitro (DIIV da proteína nãodegradadano rúmen (PNDR e os perfis de aminoácidos (AA e de ácidos graxos (AG do resíduo de cervejaria úmido (RCU e fermentado (RCF. O RCF foi obtido pelo processo de fermentação microbiana do RCU. Para determinar a DE da MS e PB do RCU e RCF, foram utilizados três novilhos da raça Holandesa, portadores de cânula ruminal. A DIIV daPNDR foi obtida pelo método de três estágios. Os dados obtidos para DE da MS e PB foram submetidos à análise de variância, em elineamento inteiramente casualizado. A fração A da PB do RCU foi de 7,9% e do RCF de 13,1% da PB. A DE da PB a 5% h-1 não diferiu (p The study evaluated the protein and carbohydrate fraction, dry matter (DM and crude protein (CP effective rumen degradability (ED, DM and CP in vitro ruminal digestibility (RDIV, rumen-undegradable protein (RUDP in vitro intestinal digestion (IDIV and amino-acid (AA and fatty acid (FA profile of the wet brewer’s grain (WBG, andfermented brewer’s grain (FBG. FBG was obtained from WBG fermentation. The DM and CP ED of WBG and FBG were determined in three Holstein steers with ruminal cannula. The IDIV of RUDP was obtained by the three-stage method. The values obtained for DM and CP ED were submitted to variance analysis, in a randomized design. The Afraction of WBG CP was 7.9%, and for FBG 13.1% of CP. The CP RD in a rate of 5% h-1 did not differ (p > 0.05 between WBG and FBG. The crude protein RDIV of FBG was 8.7% and IDIV of RUDP of WBG and FBG were of 70.5% and 72.5%, respectively. The AA and FA profile of WBG and FBG were similar. The anaerobic fermentation process did not change the nutritional characteristics of the WBG.

  8. Rumen dry matter degradability of fresh and ensiled sugarcane ...

    African Journals Online (AJOL)

    This study aimed to evaluate the chemical composition and in situ ruminal degradability of fresh (FSC) and ensiled (ESC) sugarcane. In situ dry matter degradability (DMD) was determined using the nylon bag technique with four cows equipped ruminal fistulas. Cows were fed with fresh or ensiled sugarcane and ...

  9. The effect of partial replacement of corn silage on rumen degradability, milk production and composition in lactating primiparous dairy cows

    Directory of Open Access Journals (Sweden)

    Hakan Biricik

    2010-01-01

    Full Text Available The objective of this experiment was to evaluate the effects of partial replacement of corn silage with long alfalfa hay and/or coarse chopped wheat straw on neutral detergent fibre (NDF rumen degradability, milk yield and composition in late lactating dairy cows fed diets with 50% forage on dry matter basis. Twelve late lactating Holstein primiparous cows including four cows equipped with a rumen cannula, averaging 210 ± 20 d in milk and weighing 575 ± 50 kg were randomly assigned in a 4x4 Latin square design. During each of four 21-d periods, cows were fed 4 total mixed diets that were varied in the forage sources: 1 50% corn silage (CS, 2 35% corn silage + 15% wheat straw (CSW, 3 35% corn silage + 15% alfalfa hay (CSA, 4 25% corn silage + 10% wheat straw + 15% alfalfa hay (CSWA. The production of milk averaged 18.55, 20.41 and 20.06 kg/d for unadjusted milk production, 4% fat corrected milk and solid corrected milk, respectively, and was not affected by treatments. Likewise, milk composition or production of milk components was not affected by diets and averaged 4.69% fat, 3.66% protein, 4.51% lactose, 866 g/d fat, 665 g/d protein, 824 g/d lactose. Treatments had no effect on in situ NDF soluble, degradable and potential degradability of all diets, whereas the effective degradability (ED of NDF was greater for cows fed CS diet than for cows fed CSW, CSA and CSWA diets (P<0.05. These values suggested that the partial replacement of corn silage with alfalfa hay and/or wheat straw has no unfavourable effect on the productive parameters.

  10. Proteína degradável no rúmen na dieta de bovinos: digestibilidades total e parcial dos nutrientes e parâmetros ruminais Rumen degradable protein on bovine diet: total and partial nutrient digestibility and ruminal parameters

    Directory of Open Access Journals (Sweden)

    Saul Ferreira Caldas Neto

    2008-06-01

    Full Text Available Quatro novilhos da raça Holandesa (450 kg portadores de cânula ruminal e duodenal, recebendo dietas com níveis de proteína degradável no rúmen (PDR de 50, 60, 65 e 70%, associadas a uma fonte de amido de alta degradabilidade ruminal (farinha de varredura de mandioca, foram distribuídos em quadrado latino 4 × 4 para se avaliarem as digestibilidades total e parcial dos nutrientes, a concentração de amônia e o pH ruminal. Foi utilizada, como indicador do fluxo duodenal e fecal, a cinza insolúvel em ácido. Não foi observado efeito do nível de PDR sobre o coeficiente de digestibilidade total, digestibilidade ruminal e digestibilidade intestinal da matéria seca, matéria orgânica, fibra em detergente neutro, carboidratos não-estruturais e energia bruta. O aumento do nível de PDR na dieta elevou o coeficiente de digestibilidade total e ruminal da proteína bruta e reduziu a digestibilidade intestinal desse nutriente como porcentagem do digerido. Não houve efeito dos níveis de PDR no pH ruminal, no entanto, maior concentração ruminal de amônia foi observada para as dietas com maior teor de PDR. Os resultados obtidos indicaram que o aumento no teor de PDR acarretou maior produção de nitrogênio na forma de amônia, independentemente da presença da fonte de amido de alta degradabilidade ruminal, contudo, o aporte de proteína intestinal foi semelhante para todas as dietas.Four ruminally and duodenally cannulated Holstein steers (450 kg were fed diets with rumen degradable protein (RDP levels of 50.0 60.0 65.0 and 70.0% associated with a high ruminal degradability starch (cassava by-product meal were allotted to a 4 × 4 Latin square design for the evaluation of total and partial digestibility of the nutrients, ruminal ammonia concentration and pH. The acid insoluble ash was used as a marker of the duodenal and fecal flow. No effects were observed on the level of RDP on total digestibility coefficient, ruminal digestibility and

  11. Degradation of plant wastes by anaerobic process using rumen bacteria.

    Science.gov (United States)

    Seon, J; Creuly, C; Duchez, D; Pons, A; Dussap, C G

    2003-01-01

    An operational reactor has been designed for the fermentation of a pure culture of Fibrobacter succinogenes with the constraints of strict anaerobic condition. The process is controlled by measurements of pH, redox, temperature and CO2 pressure; it allows an efficient degradation (67%) of lignocellulosic wastes such as a mixture of wheat straw, soya bean cake and green cabbage.

  12. Acid Detergent Insoluble Protein as Tropical Forages Rumen Undegradable Protein Estimator Proteína insolúvel em detergente ácido como estimador da fração protéica não degradável no rúmen de forragens tropicais

    OpenAIRE

    Lara Toledo Henriques; Hérnan Maldonado Vasques; Edenio Detmann; José Fernando Coelho da Silva; Renata Cogo Clipes; Dirlei Molinari Donatele; Ismail Ramalho Haddade

    2010-01-01

    The undegradable rumen protein fraction estimated from neutral detergent insoluble nitrogen (NDIN) was studied. A total of 540 samples were used, obtained from manual grazing simulation and esophageal extrusa in elephant grass and mombaça grass, from hays and from tropical grasses submitted to different fertilizing levels and cutting ages. The samples were analyzed for dry mater, lignin in sulfuric acid (LAS), lignin permanganate (LPER), neutral detergent insoluble protein (NDIP) and acid det...

  13. High Potential Source for Biomass Degradation Enzyme Discovery and Environmental Aspects Revealed through Metagenomics of Indian Buffalo Rumen

    Directory of Open Access Journals (Sweden)

    K. M. Singh

    2014-01-01

    Full Text Available The complex microbiomes of the rumen functions as an effective system for plant cell wall degradation, and biomass utilization provide genetic resource for degrading microbial enzymes that could be used in the production of biofuel. Therefore the buffalo rumen microbiota was surveyed using shot gun sequencing. This metagenomic sequencing generated 3.9 GB of sequences and data were assembled into 137270 contiguous sequences (contigs. We identified potential 2614 contigs encoding biomass degrading enzymes including glycoside hydrolases (GH: 1943 contigs, carbohydrate binding module (CBM: 23 contigs, glycosyl transferase (GT: 373 contigs, carbohydrate esterases (CE: 259 contigs, and polysaccharide lyases (PE: 16 contigs. The hierarchical clustering of buffalo metagenomes demonstrated the similarities and dissimilarity in microbial community structures and functional capacity. This demonstrates that buffalo rumen microbiome was considerably enriched in functional genes involved in polysaccharide degradation with great prospects to obtain new molecules that may be applied in the biofuel industry.

  14. Ruminal Degradation Characteristics and Small Intestinal Digestibility of Rumen Undegraded Protein of Six Feed Ingredients%6种饲料原料瘤胃降解特性和瘤胃非降解蛋白质的小肠消化率

    Institute of Scientific and Technical Information of China (English)

    赵连生; 牛俊丽; 徐元君; 王芳; 郑琛; 李发第; 郭江鹏; 卜登攀

    2017-01-01

    This experiment was conducted to determine the ruminal degradation characteristics and small intestinal digestibility of rumen undegraded protein (RUP) of six feed ingredients for dairy cows from Xinjiang, including corn silage, cottonseed hulls, alfalfa meal, alfalfa hay, grape seed meal and tomato sauce residue.Three lactating Holstein cows fitted with permanent rumen fistulas were selected to estimate the ruminal degradation characteristics of dry matter (DM), crude protein (CP), neutral detergent fiber (NDF) and acid detergent fiber (ADF) and small intestinal digestibility and intestinal digestible crude protein (IDCP) content of RUP by nylon-bag technique and modified three-step in vitro method.The results showed as follows:1) DM effective degradability of alfalfa meal and corn silage was higher, which was significantly higher than that of alfalfa hay and tomato sauce residue (P alfalfa meal > corn silage > alfalfa hay > cottonseed hulls > grape seed meal with significant differences among feed ingredients (P alfalfa meal > cottonseed hulls > alfalfa hay > tomato sauce residue > grape seed meal, and the differences among feed ingredients were significant (P cottonseed hulls > alfalfa hay > tomato sauce residue > alfalfa meal > grape seed meal with significant differences among feed ingredients (P苜蓿草粉>玉米青贮>苜蓿干草>棉籽壳>葡萄籽粕,各原料间差异显著(P苜蓿草粉>棉籽壳>苜蓿干草>番茄酱渣>葡萄籽粕,各组饲料原料间差异显著(P棉籽壳>苜蓿干草>番茄酱渣>苜蓿草粉>葡萄籽粕,各组饲料原料间差异显著(P0.05),显著高于依次降低的玉米青贮、番茄酱渣、葡萄籽粕、棉籽壳(P<0.05).综上所述,不同饲料原料具有不同的瘤胃降解特性,进入小肠IDCP的含量也不同.玉米青贮的DM、NDF和ADF在瘤胃的有效降解率较高,苜蓿草粉RUP的Idg较高,苜蓿草粉和苜蓿干草的IDCP含量较高.

  15. Fungal treatment of lignocellulosic biomass: Importance of fungal species, colonization and time on chemical composition and in vitro rumen degradability

    NARCIS (Netherlands)

    Kuijk, van S.J.A.; Sonnenberg, A.S.M.; Baars, J.J.P.; Hendriks, W.H.; Cone, J.W.

    2015-01-01

    The aim of this study is to evaluate fungal treatments to improve in vitro rumen degradability of lignocellulosic biomass. In this study four selective lignin degrading fungi, Ganoderma lucidum, Lentinula edodes, Pleurotus eryngii and Pleurotus ostreatus, were used to pre-treat lignocellulosic

  16. Immunization against Rumen Methanogenesis by Vaccination with a New Recombinant Protein.

    Directory of Open Access Journals (Sweden)

    Litai Zhang

    Full Text Available Vaccination through recombinant proteins against rumen methanogenesis provides a mitigation approach to reduce enteric methane (CH4 emissions in ruminants. The objective of present study was to evaluate the in vivo efficacy of a new vaccine candidate protein (EhaF on methanogenesis and microbial population in the rumen of goats. We amplified the gene mru 1407 encoding protein EhaF using fresh rumen fluid samples of mature goats and successfully expressed recombinant protein (EhaF in Escherichia coli Rosetta. This product was evaluated using 12 mature goats with half for control and other half injected with 400ug/goat the purified recombinant protein in day 1 and two subsequent booster immunizations in day 35 and 49. All measurements were undertaken from 63 to 68 days after the initial vaccination, with CH4 emissions determined using respiration calorimeter chambers. The results showed that the vaccination caused intensive immune responses in serum and saliva, although it had no significant effect on total enteric CH4 emissions and methanogen population in the rumen, when compared with the control goats. However, the vaccination altered the composition of rumen bacteria, especially the abundance of main phylum Firmicutes and genus Prevotella. The results indicate that protein EhaF might not be an effective vaccine to reduce enteric CH4 emissions but our vaccine have potential to influence the rumen ecosystem of goats.

  17. Comparison of organic matter degradation in several feedstuffs in the rumen as determined with the nylon bag and gas production techniques

    NARCIS (Netherlands)

    Cone, John W.; Van Gelder, Antonie H.; Bachmann, Herwig; Hindle, Vincent A.

    2002-01-01

    Organic matter (OM) degradation of 21 feedstuffs was investigated with rumen fluid using a rumen in situ technique and a gas production technique. Fitting the nylon bag data to an exponential model showed that there was a high variation in the rate of OM degradation ranging from 1.7% h-1 for

  18. Feeding, evaluating, and controlling rumen function.

    Science.gov (United States)

    Lean, Ian J; Golder, Helen M; Hall, Mary Beth

    2014-11-01

    Achieving optimal rumen function requires an understanding of feeds and systems of nutritional evaluation. Key influences on optimal function include achieving good dry matter intake. The function of feeds in the rumen depends on other factors including chemical composition, rate of passage, degradation rate of the feed, availability of other substrates and cofactors, and individual animal variation. This article discusses carbohydrate, protein, and fat metabolism in the rumen, and provides practical means of evaluation of rations in the field. Conditions under which rumen function is suboptimal (ie, acidosis and bloat) are discussed, and methods for control examined. Copyright © 2014 Elsevier Inc. All rights reserved.

  19. Molecular spectroscopic features of protein in newly developed chickpea: Relationship with protein chemical profile and metabolism in the rumen and intestine of dairy cows

    Science.gov (United States)

    Sun, Baoli; Khan, Nazir Ahmad; Yu, Peiqiang

    2018-05-01

    The first aim of this study was to investigate the nutritional value of crude protein (CP) in CDC [Crop Development Centre (CDC), University of Saskatchewan] chickpea varieties (Frontier kabuli and Corinne desi) in comparison with a CDC barley variety in terms of: 1) CP chemical profile and subfractions; (2) in situ rumen degradation kinetics and intestinal digestibility of CP; 2) metabolizable protein (MP) supply to dairy cows; and (3) protein molecular structure characteristics using advanced molecular spectroscopy. The second aim was to quantify the relationship between protein molecular spectral characteristics and CP subfractions, in situ rumen CP degradation characteristics, intestinal digestibility of CP, and MP supply to dairy cows. Samples (n = 4) of each variety, from two consecutive years were analyzed. Chickpeas had higher (P content (21.71-22.11 vs 12.96% DM), with higher (P content, and any of the measured in situ degradation and molecular spectral characteristics of protein. The content of RUP was positively (r = 0.94, P content of CP (R2 = 0.91) D-fraction (R2 = 0.82), RDP (R2 = 0.77), RUP (R2 = 0.77), TDP (R2 = 0.98), MP (R2 = 0.80), and FMV (R2 = 0.80) can be predicted from amide II peak height. Despite extensive ruminal degradation, chickpea is a good source of MP for dairy cows, and molecular spectroscopy can be used to rapidly characterize feed protein molecular structures and predict their digestibility and nutritive value.

  20. QUANTIFICATION OF THE EFFICIENCY OF RUMEN MICROBIAL PROTEIN SYNTHESIS IN STEERS FED GREEN TROPICAL GRASS

    Directory of Open Access Journals (Sweden)

    MARTHEN L. MULLIK

    2012-09-01

    Full Text Available ABSTRACT The rate of rumen microbial crude protein (MCP supply to the intestines is a crucial element in the current rumen models to predict respond of ruminants to a certain diet. Data from tropical pastures always below predicted results from the existing rumen models. Thus, quantification of the rumen MCP supply from tropical grass will improve predictive rate under tropical feeding conditions. Four Brahman crossbred steers (457±20.1 kg were used in a metabolism study. Pangola grass (Digitaria erianthe cv. Steudal was harvested every morning and fed to the animals soon after. Parameters measured were EMPS, intake, fractional passage rates, and rumen ammonia concentration. The EMPS was estimated using purine derivative excretion in urine. Crude protein and water soluble carbohydrates content were 6.3 and 7.4% of dry matter (DM respectively. DM intake was 1.6% live weight. Average rumen ammonia concentration was 69 mg/L whilst rumen passage rates were 7.84 and 6.92 %/h for fluid and solids respectively. EMPS was only 72 g MCP/kg digestible organic matter. It might be concluded that EMPS in steers consuming green pangola grass was below the minimum level for forage diets adopted in the current feeding standards. ABSTRAK Tingkat pasokan protein mikroba rumen (MCP ke usus halus merupakan salah satu unsur kunci dalam meramal respon pertumbuhan ruminan terhadap ransum tertentu. Data MCP hijauan tropis selalu berada di bawah nilai prediksi model rumen yang dipakai saat ini. Dengan demikian, kuantifikasi pasokan MCP rumput tropis diharapkan menjadi masukan untuk meningkatkan kemampuan prediksi model rumen untuk pakan daerah tropis. Empat sapi jantan muda Brahman persilangan (457±20,1 kg digunakan dalam sebuah penelitian metabolisme. Rumput pangola (Digitaria erianthe cv. Steudal dipanen setiap pagi dan langsung diberikan kepada ternak dalam kandang metabolis. Parameter yang diukur adalah produksi MCP dan efisiensi sintesis MCP (Emps, konsumsi, laju

  1. Truly Absorbed Microbial Protein Synthesis, Rumen Bypass Protein, Endogenous Protein, and Total Metabolizable Protein from Starchy and Protein-Rich Raw Materials

    NARCIS (Netherlands)

    Parand, Ehsan; Vakili, Alireza; Mesgaran, Mohsen Danesh; Duinkerken, Van Gert; Yu, Peiqiang

    2015-01-01

    This study was carried out to measure truly absorbed microbial protein synthesis, rumen bypass protein, and endogenous protein loss, as well as total metabolizable protein, from starchy and protein-rich raw feed materials with model comparisons. Predictions by the DVE2010 system as a more

  2. Perlindungan Protein Menggunakan Tanin dan Saponin Terhadap Daya Fermentasi Rumen dan Sintesis Protein Mikrob (PROTECTION OFPROTEINUSINGTANNINS AND SAPONINS OF RUMEN DIGESTIBILITYAND MICROBESSYNTHESISPROTEIN

    Directory of Open Access Journals (Sweden)

    Ana Shofi Ani

    2015-10-01

    Full Text Available The objective of this experiment was to examine protection of protein using tannins and saponins toimprove rumen digestibility and microb-mediated protein synthesis in vitro. Rumen fluids used as inoculumwas collected from a composite of two female adult fistulatedongole cattle weighed of ±300 kg with theage of five years old. The experimental design used in this study was a completely randomized design withsix treatments and three replication of each treatment. The six treatments consisted of T0: Proteinconcentrates without protection, T1: protein concentrates protected with 1.2% saponin, T2: proteinconcentrates protected with 0.5% tannin and 0.9% saponin , T3: protein concentrates protected with 1.0%tannin and 0.6% saponin, T4: protein concentrates protected with 1.5 % tannin and 0.3% saponin and T5:protein concentrates protected with 2.0% tannins. The result showed that treatment with tannin, saponinand their combination had a significantly affect (P<0,05 on the level of ammonia (NH3, the total volatilefatty acids (VFA, and total protein. Protection of proteins with combination of 1,0% tannin and 0.6%saponin resulted in best effect on feed protein as shown by its NH3 concentration, total VFA and totalprotein. This indicates the level of protection of feed protein can improve rumen digestibility and microbesmediatedprotein synthesis, as showed in the concentration of N-NH3, total VFA and total protein.

  3. The Concentrations of Rumen Fluid Volatile Fatty Acids and Ammonia, and Rumen Microbial Protein Production in Sheep Given Feed During the Day and Night Time

    Science.gov (United States)

    Gumilar, D. A. K. W.; Rianto, E.; Arifin, M.

    2018-02-01

    An experimental study was carried out to investigate the concentrations of volatile (VFA), ammonia and microbial protein production of rumen fluid in sheep given fedd during the day and at night. This study used 12 fat-tailed rams aged 12-18 months and weighed 24,12 ± 25 kg (CV = 10,51%). The rams were fed a complete feed containing 16.64% protein and 68,33% total digestible nutrients (TDN). The rams were allocated into a completely randomised design with 3 treatments and 4 replications. The treatments applied were: T1: day time feeding (6.00 hrs - 18.00 hrs); T2: night time feeding (18.00 hrs - 6.00 hrs); and T3: day and night time feedings (6.00 hrs - 6.00 hrs). The parameters observed were dry matter intake (DMI), rumen VFA concentration, rumen ammonia concentration, rumen rmicrobial protein production and the efficiency of rumen microbial protein production. The results showed that feeding time did not significantly affect (P>0.05) all the parameters observed. Dry matter intake, VFA concentration, ammonia concentration, the microbial protein production of rumen fluid and the efficiency of microbial protein production were 1,073g/d, 49.69 mmol; 4.77 mg N/100 ml, 12,111 g/d and 19.96 g per kg digestible organic matter intake (DOMI), respectively. It is concluded that feeding time did not affect DMI, condition of rumen fluid and rumen microbial protein production in sheep.

  4. In situ Rumen Degradation Kinetics of High-Protein Forage Crops in Temperate Climates Cinética de Degradación Ruminal in situ en Forrajes de Alto Contenido Proteico en Clima Templado

    Directory of Open Access Journals (Sweden)

    Ximena Valderrama L.

    2011-12-01

    Full Text Available The present study was conducted to evaluate the nutritional value and in situ degradation kinetics of eight high protein forage crops: alfalfa (Medicago sativa L., forage oat (Avena sativa L., mixed pasture, and ryegrass (Lolium multiflorum Lam. pasture in early vegetative stages, two forage lupins (Lupinus albus L. in early bloom stages, sugar beet (Beta vulgaris L. and kale (Brassica napus var. pabularia (DC. Rchb. leaves at root maturity. Dry matter (DM and crude protein (CP degradation kinetics were evaluated by the nylon bag technique through the in situ procedure described by 0rskov and MacDonald (1979 using three ruminally cannulated sheep. Chemical composition of the forage crops showed on average 13.7% DM; 21.4% CP; 31.5% neutral detergent fiber (NDF; 17.7% crude fiber (CF, 80.6% digestibility of organic matter (DOMD and 12.13 MJ kg-1 metabolizable energy (ME. The high total degradability of forage crops reported here (> 87% DM; > 93% CP can be associated with the presence of large quantities of fraction a (> 34% DMa; > 29% CPa and high degradability of fraction b, resulting in low amounts of undegradable fraction (U (7.02% DM and 3.55% CP. Correlations between CPb and DMb degradability (r = 0.79 and CPc and DMc degradation rates (r = 0.78 were high, however differences in c were not explained by differences in CP or NDF contents, nor by the amounts of a or b fractions. Degradation for DM and CP during the first 6 h of incubation was strongly and inversely correlated to b (36 h (r = 0.93 (P El presente estudio se desarrolló con el objetivo de evaluar el valor nutricional y la cinética de degradación in situ de ocho forrajes de alto valor proteico: alfalfa (Medicago sativa L., avena (Avena sativa L., pastos mixtos y pastos de ballica (Lolium multiflorum Lam., en las primeras etapas vegetativas, dos lupinos forrajeros (Lupinus albus L. en etapas inicio de la floración, hojas de remolacha azucarera (Beta vulgaris L. y de col (Brassica

  5. Molecular spectroscopic features of protein in newly developed chickpea: Relationship with protein chemical profile and metabolism in the rumen and intestine of dairy cows.

    Science.gov (United States)

    Sun, Baoli; Khan, Nazir Ahmad; Yu, Peiqiang

    2018-05-05

    The first aim of this study was to investigate the nutritional value of crude protein (CP) in CDC [Crop Development Centre (CDC), University of Saskatchewan] chickpea varieties (Frontier kabuli and Corinne desi) in comparison with a CDC barley variety in terms of: 1) CP chemical profile and subfractions; (2) in situ rumen degradation kinetics and intestinal digestibility of CP; 2) metabolizable protein (MP) supply to dairy cows; and (3) protein molecular structure characteristics using advanced molecular spectroscopy. The second aim was to quantify the relationship between protein molecular spectral characteristics and CP subfractions, in situ rumen CP degradation characteristics, intestinal digestibility of CP, and MP supply to dairy cows. Samples (n=4) of each variety, from two consecutive years were analyzed. Chickpeas had higher (Pmolecular spectral data of chickpeas can be distinguished from the barley. The two chickpeas did not differ in CP content, and any of the measured in situ degradation and molecular spectral characteristics of protein. The content of RUP was positively (r=0.94, Pmolecular spectroscopy can be used to rapidly characterize feed protein molecular structures and predict their digestibility and nutritive value. Crown Copyright © 2018. Published by Elsevier B.V. All rights reserved.

  6. In Vitro Digestibilities of Six Rumen Protected Fat-Protein Supplement Formulas

    Directory of Open Access Journals (Sweden)

    Lilis Hartati

    2012-01-01

    Full Text Available Abstract. The aim of the research was to evaluate the efficacy of protection method of rumen protected fat-protein supplements. In vitro digestibility test was carried out to examine nutrients digestibility of different supplement formula based on the sources of protein and oil. The research used two sources of fat namely crude palm oil (CPO and fish oil (FO and three sources of protein namely milk skim, soy flour, and soybean meal. Thus there were 6 combinations that subjected in the in vitro digestibility test. The observed variables were the digestibility of dry matter (DM, organic matter (OM, crude fat (CF, and crude protein (CP. Results indicated that the method for protecting protein and fat was effective. This was showed by low nutrients digestibility in the rumen and high nutrients digestibility in the post rumen. In conclusion the combination between skim milk and CPO gave the best results among the other supplement formula. Keywords: rumen protected nutrient, fat-protein supplement, rumen digestibility, in vitro Animal Production 14(1:1-5, January 2012

  7. The effect of lime pre-treatments of date palm leaves on delignification and in vitro rumen degradability

    NARCIS (Netherlands)

    Ghorbani, M.; Ahmadi, F.; Rajaee Rad, A.; Zamiri, M.J.; Cone, J.W.; Polikarpov, I.

    2017-01-01

    Experiments were conducted to determine the effect of lime pre-treatment on the chemical composition and in vitro rumen degradability of date palm leaves (DPL). Lime pre-treatments, with or without oxygen supply, were applied for 1, 2 and 3 weeks at 25 and 40 °C. Lime was neutralized by the

  8. Starch degradation in rumen fluid as influenced by genotype, climatic conditions and maturity stage of maize, grown under controlled conditions

    NARCIS (Netherlands)

    Ali, M.; Cone, J.W.; Hendriks, W.H.; Struik, P.C.

    2014-01-01

    Starch is the major component of maize kernels, contributing significantly to the feeding value of forage maize when fed to ruminants. The effects of genotype, climatic conditions and maturity stage on starch content in the kernels and on in vitro starch degradability in rumen fluid were

  9. Shifts in the rumen microbiota due to the type of carbohydrate and level of protein ingested by dairy cattle are associated with changes in rumen fermentation.

    Science.gov (United States)

    Belanche, Alejandro; Doreau, Michel; Edwards, Joan E; Moorby, Jon M; Pinloche, Eric; Newbold, Charles J

    2012-09-01

    Balancing energy and nitrogen in the rumen is a key to both profitability and environmental sustainability. Four dairy cows were used in a Latin square experimental design to investigate the effect of severe nitrogen underfeeding (110 vs. 80% of requirements) and the type of carbohydrate consumed [neutral detergent fiber rich (FIB) vs. starch rich (STA)] on the rumen ecosystem. These dietary treatments modified both rumen fermentation and microbial populations. Compared with STA diets, consumption of FIB diets increased bacterial and fungal diversity in the rumen and also increased the concentrations of cellulolytic microorganisms, including protozoa (+38%), anaerobic fungi (+59%), and methanogens (+27%). This microbial adaptation to fiber utilization led to similar digestibility values for the 2 carbohydrate sources and was accompanied by a shift in the rumen fermentation patterns; when the FIB diets were consumed, the cows had greater ruminal pH, ammonia concentrations, and molar proportions of acetate and propionate compared with when they consumed the STA diets. Certain rumen microorganisms were sensitive to a shortage of nitrogen; rumen concentrations of ammonia were 49% lower when the low-protein (LP) diets were consumed as were total bacteria (-13%), anaerobic fungi (-28%), methanogens (-27%), protozoa (-19%), cellulolytic bacteria, and microbial diversity compared with when the high-protein (HP) diets were consumed. As a result, the digestibility of the LP diets was less than that of the HP diets. These findings demonstrated that the rumen microbial ecosystem is directly linked to the rumen fermentation pattern and, to some extent, to the efficiency of diet utilization by dairy cattle.

  10. Intestinal digestibility of amino acids in rumen-undegraded protein estimated using a precision-fed cecectomized rooster bioassay: II. Distillers dried grains with solubles and fish meal.

    Science.gov (United States)

    Boucher, S E; Calsamiglia, S; Parsons, C M; Stein, H H; Stern, M D; Erickson, P S; Utterback, P L; Schwab, C G

    2009-12-01

    The objectives of this experiment were to measure intestinal digestibility of AA in the rumen-undegraded protein fraction (RUP-AA) of distillers dried grains with solubles (DDGS) and fish meal (FM) samples and to determine whether these feeds contain a constant protein fraction that is undegradable in the rumen and indigestible in the small intestine, as assumed in the French Institut National de la Recherche Agronomique (Paris, France) and Scandinavian AAT-PBV (AAT = AA absorbed from small intestine; PBV = protein balance in the rumen) models. Five sources of DDGS and 5 sources of FM were obtained from Feed Analysis Consortium, Inc. (Champaign, IL). To obtain the rumen-undegradable protein fraction, samples were ruminally incubated in situ for 16 h in 4 lactating cows, and the collected rumen-undegraded residues (RUR) were pooled by sample. Subsamples of the intact feeds and RUR were crop-intubated to 4 cecectomized roosters, and total excreta were collected for 48 h. Intact feeds, RUR, and excreta were analyzed for AA. Basal endogenous AA loss estimates were obtained from fasted birds and were used to calculate standardized digestibility of RUP-AA and AA in the intact feeds. Indigestibility coefficients of the intact feeds were calculated as (100 - % standardized AA digestibility), and indigestibility of the RUR was calculated as [(100 - % ruminal degradation of AA) x (100 - % standardized RUP-AA digestibility)/100]. Results indicate that standardized digestibility of feed-AA differs from RUP-AA for DDGS samples but not for FM samples, and that standardized digestibility of individual AA differs within samples. For the DDGS samples, standardized feed-AA and RUP-AA digestibility values were most often lowest for His and Lys and highest for Met and Trp. For FM samples, standardized feed-AA and RUP-AA digestibility values were most often lowest for His and highest for Trp. Results also indicate that DDGS and most FM samples do not contain a constant protein fraction

  11. ESTIMACIÓN DE LA DEGRADABILIDAD EFECTIVA EN EL RUMEN MEDIANTE MÉTODOS NUMÉRICOS ESTIMATION OF EFFECTIVE DEGRADABILITY IN RUMEN TROUGH NUMERICAL METHODS

    Directory of Open Access Journals (Sweden)

    Héctor Jairo Correa Cardona

    2008-12-01

    Full Text Available La estimación correcta de la degradabilidad ruminal efectiva en el rumen (DE de las distintas fracciones nutricionales de los alimentos es fundamental para el desarrollo de programas eficientes de alimentación para rumiantes. La propuesta clásica para estimar la degradabilidad efectiva en el rumen de la fracción potencialmente degradable (DEb=bKd/(Kd+Kp, no es correcta ya que se basa en la presunción de que las constantes de la cinética de la degradabilidad (Kd y del pasaje ruminal (Kp representan la velocidad de fermentación y de pasaje ruminal, respectivamente cuando estas realmente representan la relación constante entre la aceleración y la velocidad de degradación y de pasaje. Se presenta, entonces, una nueva propuesta coherente con las bases matemáticas de la cinética de la degradación y el pasaje ruminal de la fracción potencialmente degradable en el rumen (b que requiere el uso de métodos numéricos para despejar el tiempo “t” de la expresión 1 = e-kd*t + e-kp*t, que al reemplazarlo en la expresión b*e-kd*t, permite calcular la DEb. La estimación de la DEb por este método permite obtener datos confiables y coherentes con las bases matemáticas de la cinética de la degradación y el pasaje ruminal de las fracciones nutricionales.A correct estimation of effective degradability in rumen of nutritional fractions of feedstuff is basic to develop efficient feed programs to ruminants. However, the classic proposal to estimate the effective degradability in rumen of potentially degradable fraction (EDb=bKd/(Kd+Kp, is not correct since it is based on the presumption that the constants of the kinetics of the ruminal degradability (Kd and passage (Kp represent the speed of the ruminal fermentation and passage, respectively, when these really represent the constant relationship between the acceleration and speed of degradation and of passage. In this paper is then proposed a new mathematical procedure coherent with the

  12. Impact of levels of total digestible nutrients on microbiome, enzyme profile and degradation of feeds in buffalo rumen.

    Science.gov (United States)

    Kala, Anju; Kamra, D N; Kumar, Avinash; Agarwal, Neeta; Chaudhary, L C; Joshi, C G

    2017-01-01

    The present study was aimed at understanding a shift in rumen microbiome of buffaloes fed various levels of total digestible nutrients. To understand the process, the metagenomics of rumen microbes, in vivo and in vitro rumen fermentation studies were carried out. Three rumen fistulated adult male Murrah buffaloes were fed three isonitrogenous diets varying in total digestible nutrients (70, 85 and 100% of TDN requirement) in 3X3 switch over design. On dry matter basis, wheat straw/ roughage content were 81, 63 and 51% and that of maize grain was 8, 16 and 21% in three diets respectively. After 20 d of feeding, rumen liquor and rumen contents were sampled just before (0h) and 4h post feeding. Ruminococcus flavefaciens and R. albus (estimated with real time PCR) were higher in high roughage diets. The predominant phyla in all the three groups were Bacteroidetes, Firmicutes followed by Proteobacteria, Actinobacteria and Fibrobacteres. A core group of more than fifty rumen bacteria was present in all the animals with very little variations due to level of TDN. The most predominant bacterial genera reported in order of decreasing abundance were: Prevotella, Bacteroides, Clostridium, Ruminococcus, Eubacterium, Parabacteroides, Fibrobacter, Butyrivibrio etc. The higher diversity of the enyzmes families GH 23, GH 28, GH 39, GH 97, GH 106, and GH 127 (the enzymes active in fibre and starch degradation) were significantly higher on 100%TDN diet while CE 14 (required for the hydrolysis of bond between carbohydrate and lignin) was higher on low TDN (70%) diet, indicating ester bond cleavage was better in animals fed high roughage (wheat straw) diet.

  13. What happens in the bag? : development and evaluation of a modified in situ protocol to estimate degradation of nitrogen and starch in the rumen

    NARCIS (Netherlands)

    Jonge, de L.H.

    2015-01-01

    The most widely used method to estimate the rumen degradation of dietary components in feedstuffs is the in situ or in sacco method. This method is based on rumen incubation of substrate (feed) in nylon or dacron bags followed by rinsing and analysis of the residue. Small pores

  14. Metagenome Analysis of Protein Domain Collocation within Cellulase Genes of Goat Rumen Microbes

    Directory of Open Access Journals (Sweden)

    SooYeon Lim

    2013-08-01

    Full Text Available In this study, protein domains with cellulase activity in goat rumen microbes were investigated using metagenomic and bioinformatic analyses. After the complete genome of goat rumen microbes was obtained using a shotgun sequencing method, 217,892,109 pair reads were filtered, including only those with 70% identity, 100-bp matches, and thresholds below E−10 using METAIDBA. These filtered contigs were assembled and annotated using blastN against the NCBI nucleotide database. As a result, a microbial community structure with 1431 species was analyzed, among which Prevotella ruminicola 23 bacteria and Butyrivibrio proteoclasticus B316 were the dominant groups. In parallel, 201 sequences related with cellulase activities (EC.3.2.1.4 were obtained through blast searches using the enzyme.dat file provided by the NCBI database. After translating the nucleotide sequence into a protein sequence using Interproscan, 28 protein domains with cellulase activity were identified using the HMMER package with threshold E values below 10−5. Cellulase activity protein domain profiling showed that the major protein domains such as lipase GDSL, cellulase, and Glyco hydro 10 were present in bacterial species with strong cellulase activities. Furthermore, correlation plots clearly displayed the strong positive correlation between some protein domain groups, which was indicative of microbial adaption in the goat rumen based on feeding habits. This is the first metagenomic analysis of cellulase activity protein domains using bioinformatics from the goat rumen.

  15. Temporal metagenomic and metabolomic characterisation of fresh perennial ryegrass degradation by rumen bacteria

    Directory of Open Access Journals (Sweden)

    Olga Mayorga

    2016-11-01

    Full Text Available Understanding the relationship between ingested plant material and the attached microbiome is essential for developing methodologies to improve ruminant nutrient use efficiency. We have previously shown that perennial ryegrass (PRG rumen bacterial colonisation events follow a primary (up to 4 h and secondary (after 4 h pattern based on the differences in diversity of the attached bacteria. In this study we investigated temporal niche specialisation of primary and secondary populations of attached rumen microbiota using metagenomic shotgun sequencing as well as monitoring changes in the plant chemistry using mid-infrared spectroscopy (FT-IR. Metagenomic Rapid Annotation using Subsystem Technology (MG-RAST taxonomical analysis of shotgun metagenomic sequences showed that the genera Butyrivibrio, Clostridium, Eubacterium, Prevotella and Selenomonas dominated the attached microbiome irrespective of time. MG-RAST also showed that Acidaminococcus, Bacillus, Butyrivibrio and Prevotella rDNA increased in read abundance during secondary colonisation, whilst Blautia decreased in read abundance. MG-RAST Clusters of Orthologous Groups (COG functional analysis also showed that the primary function of the attached microbiome was categorised broadly within ‘metabolism’; predominantly amino acid, carbohydrate, and lipid metabolism and transport. Most sequence read abundances (51.6, 43.8, and 50.0% of COG families pertaining to amino acid, carbohydrate and lipid metabolism, respectively within these categories were higher in abundance during secondary colonisation. Kyoto encyclopedia of genes and genomes (KEGG pathways analysis confirmed that the PRG- attached microbiota present at 1 and 4 h of rumen incubation possess a similar functional capacity, with only a few pathways being uniquely found in only one incubation time point only. FT-IR data for the plant residues also showed that the main changes in plant chemistry between primary and secondary

  16. Reaction of some rumen micro flora to different supplementary feeds ...

    African Journals Online (AJOL)

    Ruminant animals lack enzymes to break down fibrous feeds but they harbor microorganisms capable of degrading their feeds. Rumen microbes are affected by feed substrates. The purpose of this study was to evaluate rumen microbial changes as the function of varying supplementary feeds. Two protein supplements ...

  17. Role of anaerobic fungi in wheat straw degradation and effects of plant feed additives on rumen fermentation parameters in vitro.

    Science.gov (United States)

    Dagar, S S; Singh, N; Goel, N; Kumar, S; Puniya, A K

    2015-01-01

    In the present study, rumen microbial groups, i.e. total rumen microbes (TRM), total anaerobic fungi (TAF), avicel enriched bacteria (AEB) and neutral detergent fibre enriched bacteria (NEB) were evaluated for wheat straw (WS) degradability and different fermentation parameters in vitro. Highest WS degradation was shown for TRM, followed by TAF, NEB and least by AEB. Similar patterns were observed with total gas production and short chain fatty acid profiles. Overall, TAF emerged as the most potent individual microbial group. In order to enhance the fibrolytic and rumen fermentation potential of TAF, we evaluated 18 plant feed additives in vitro. Among these, six plant additives namely Albizia lebbeck, Alstonia scholaris, Bacopa monnieri, Lawsonia inermis, Psidium guajava and Terminalia arjuna considerably improved WS degradation by TAF. Further evaluation showed A. lebbeck as best feed additive. The study revealed that TAF plays a significant role in WS degradation and their fibrolytic activities can be improved by inclusion of A. lebbeck in fermentation medium. Further studies are warranted to elucidate its active constituents, effect on fungal population and in vivo potential in animal system.

  18. Effect of heat treatment on in situ rumen degradability and in vitro ...

    African Journals Online (AJOL)

    fat soyabean (FFSB) and solvent extracted soyabean meal (SBM) on the in situ dry matter (DM) and protein degradability, and in vitro gas production kinetics of the protein sources. Ruminal disappearance of DM and crude protein (CP), and in ...

  19. Protein-energy supplementation for lambs: feed intake, ingestive behavior, rumen parameters and nutrient digestibility

    Directory of Open Access Journals (Sweden)

    Pâmila Carolini Gonçalves da Silva

    2017-08-01

    Full Text Available The study evaluated the effects of dietary protein-energy supplementation on feed intake, ingestive behavior, rumen parameters and nutrient digestibility in lambs. Four castrated lambs with 31.9 kg mean body weight and fistulated rumen were tested. distributed into latin square design (4x4, four treatments were tested over four periods of time: no supplementation (control or with supplementation at 8, 16 and 24 g kg-1 body weight. The supplement (soybean meal, soybean hulls, ground corn and minerals was provided with roughage (Tifton Bermudagrass, Cynodon spp., hay, which was offered ad libitum once a day, at 8h00. In treatments receiving 0 (control, 8, 16 and 24 g kg-1 supplementation, dry matter intake was 685.26, 742.86, 842.51 and 1013.33 g day-1, crude protein intake was 80.18, 95.98, 118.64, 150.14 g day-1 and metabolizable energy intake 1.55, 1.91, 2.31 and 2.98 g day-1, respectively. Treatments receiving the highest supplementation levels spent less time with rumination and feeding and rested for longer (P < 0.05. Protein-energy supplementation level did not affect rumen parameters. Average rumen pH was 6.3 and rumen ammonia nitrogen 165 mg dL-1; both were affected by sampling time. Supplementation levels until 24 g kg-1 BW improves feed intake and nutrient digestibility linearly and changes ingestive behavior, lowering rumination time without affecting rumen parameters.

  20. Amino acid profiles of rumen undegradable protein: a comparison between forages including cereal straws and alfalfa and their respective total mixed rations.

    Science.gov (United States)

    Wang, B; Jiang, L S; Liu, J X

    2018-06-01

    Optimizing the amino acid (AA) profile of rumen undegradable protein (RUP) can positively affect the amount of milk protein. This study was conducted to improve knowledge regarding the AA profile of rumen undegradable protein from corn stover, rice straw and alfalfa hay as well as the total mixed ratio diets (TMR) based on one of them as forage source [forage-to-concentrate ratio of 45:55 (30% of corn stover (CS), 30% of rice straw (RS), 23% of alfalfa hay (AH) and dry matter basis)]. The other ingredients in the three TMR diets were similar. The RUP of all the forages and diets was estimated by incubation for 16 hr in the rumen of three ruminally cannulated lactating cows. All residues were corrected for microbial colonization, which was necessary in determining the AA composition of RUP from feed samples using in situ method. Compared with their original AA composition, the AA pattern of forages and forage-based diets changed drastically after rumen exposure. In addition, the extent of ruminal degradation of analysed AA was not constant among the forages. The greatest individual AA degradability of alfalfa hay and corn stover was Pro, but was His of rice straw. A remarkable difference was observed between microbial attachment corrected and uncorrected AA profiles of RUP, except for alfalfa hay and His in the three forages and TMR diets. The ruminal AA degradability of cereal straws was altered compared with alfalfa hay but not for the TMR diets. In summary, the AA composition of forages and TMR-based diets changed significantly after ruminal exposure, indicating that the original AA profiles of the feed cannot represent its AA composition of RUP. The AA profile of RUP and ruminal AA degradability for corn stover and rice straw contributed to missing information in the field. © 2017 Blackwell Verlag GmbH.

  1. Rumen metabolism and recent developments

    International Nuclear Information System (INIS)

    Smith, R.H.; Oldham, J.D.

    1983-01-01

    The rumen contains a heterogenous microbial population unevenly distributed in the fluid, in solid fractions of the contents and on the rumen wall. Understanding the ways in which these populations interact with dietary nutrients or those of endogenous origin is essential for improving prediction of nutritional responses, for developing economic production systems and for making maximum use of low quality feeds. Important factors affecting fibre digestion, protein degradation and microbial protein synthesis include turnover rates of individual digesta fractions, long lasting or transient deficiencies or excesses of specific nutrients, and the susceptibilities to microbial attack of naturally occurring or treated dietary components. (author)

  2. Comparison of different protein sources in enriched grain mixture for ...

    African Journals Online (AJOL)

    -inname voorsien. ..... Animal nutrition. Longman, New York. MERCER, lR., ALLEN, S.A. & MILLER, EL, 1980. Rumen bacterial protein synthesis and the proportion of dietary protein escaping degradation in the rumen of sheep. Br. J. NUlr.

  3. Molecular basis of structural makeup of hulless barley in relation to rumen degradation kinetics and intestinal availability in dairy cattle: A novel approach.

    Science.gov (United States)

    Damiran, D; Yu, P

    2011-10-01

    To date, no study has been done of molecular structures in relation to nutrient degradation kinetics and intestinal availability in dairy cattle. The objectives of this study were to (1) reveal molecular structures of hulless barley affected by structural alteration using molecular spectroscopy (diffuse reflectance infrared Fourier transform) as a novel approach, and (2) quantify structure features on a molecular basis in relation to digestive kinetics and nutritive value in the rumen and intestine in cattle. The modeled feeds in this study were 4 types of hulless barley (HB) cultivars modified in starch traits: (a) normal starch cultivar, (b) zero-amylose waxy, (c) waxy, and (d) high-amylose. The molecular structural features were determined using diffuse reflectance infrared Fourier transform spectroscopy in the mid-infrared region (ca. 4,000-800 cm(-1)) of the electromagnetic spectrum. The items assessed included infrared intensity attributed to protein amide I (ca. 1,715-1,575 cm(-1)), amide II (ca. 1,575-1,490 cm(-1)), α-helix (ca. 1,648-1,660 cm(-1)), β-sheet (ca. 1,625-1,640 cm(-1)), and their ratio, β-glucan (ca. 1,445-1,400 cm(-1)), total carbohydrates (CHO; ca. 1,188-820 cm(-1)) and their 3 major peaks, structural carbohydrates (ca. 1,277-1,190 cm(-1)), and ratios of amide I to II and amide I to CHO. The results show that (1) the zero-amylose waxy was the greatest in amide I and II peak areas, as well as in the ratio of protein amide I to CHO among HB; (2) α-helix-to-β-sheet ratio differed among HB: the high-amylose was the greatest, the zero-amylose waxy and waxy were the intermediate, and the normal starch was the lowest; (3) HB were similar in β-glucan and CHO molecular structural makeup; (4) altered starch HB cultivars were similar to each other, but were different from the normal starch cultivar in protein molecular makeup; and (5) the rate and extent of rumen degradation of starch and protein were highly related to the molecular structural

  4. Intestinal digestibility of amino acids in rumen undegradable protein estimated using a precision-fed cecectomized rooster bioassay: I. Soybean meal and SoyPlus.

    Science.gov (United States)

    Boucher, S E; Calsamiglia, S; Parsons, C M; Stein, H H; Stern, M D; Erickson, P S; Utterback, P L; Schwab, C G

    2009-09-01

    The objectives of this experiment were to measure intestinal digestibility of AA in rumen undegradable protein (RUP-AA) in soybean meal (SBM) and expeller SBM (SoyPlus, West Central, Ralston, IA; SP) and to determine if these feeds contain a constant protein fraction that is undegradable in the rumen and indigestible in the small intestine, as assumed in the French Institut National de la Recherche Agronomique (Paris, France) and Scandinavian AAT-PBV (AAT = AA absorbed from small intestine; PBV = protein balance in the rumen) models. Three samples of SBM and 3 samples of SP were obtained from the Feed Analysis Consortium Inc. (Savoy, IL). To obtain the RUP fraction, samples were ruminally incubated in situ for 16 h in 4 lactating cows, and the collected rumen undegraded residues (RUR) were pooled by sample. Subsamples of the intact feeds and RUR were crop intubated to 4 cecectomized roosters, and total excreta were collected for 48 h. Intact feeds, RUR, and excreta were analyzed for AA. Basal endogenous AA loss estimates were obtained from fasted birds and were used to calculate standardized digestibility of AA in the intact feeds and RUP-AA. Indigestibility coefficients of the intact feeds were calculated as (100 - % standardized AA digestibility), and indigestibility of the RUR was calculated as [(100 - % ruminal degradation of AA) x [(100 - % standardized RUP-AA digestibility)]/100]. Results indicated that standardized digestibility of feed-AA was similar to standardized digestibility of RUP-AA for SBM and SP samples and that standardized digestibility of individual AA differed within samples. Standardized feed-AA and RUP-AA digestibility values were lowest for Lys and Cys and highest for Trp and Met. Results also indicated that SBM and SP did not contain a constant protein fraction that was both undegradable in the rumen and indigestible in the small intestine. Indigestibility values of RUR were lower than in intact feeds, suggesting that SBM and SP contain a

  5. Measurements of carbon dioxide production rates in the rumen of buffalo-calves fed on two levels of crude proteins

    International Nuclear Information System (INIS)

    Varma, A.; Verma, D.N.; Singh, U.B.; Ranjhan, S.K.; Agarwal, Ranjana

    1974-01-01

    The production rates of carbon dioxide in the rumen of buffalo calves have been measured by single injection isotope dilution technique. One group of calves received 11 percent less proteins and the other 20 percent more than that recommended by the NRC. About 258 Ci of NaH 1 CO 3 was injected in a single dose into the rumen through a cannula and mixed manually with the rumen contents. Samples of the rumen liquor were drawn for 560 min and were analysed for the specifiradioactivity of carbon dioxide. The decline in the specific radioactivity as a function of time was fitted to an equation. The dilution curves were described by a sum of 2 exponential components. Mathematical equations were used to estimate the total CO 2 entry rates in the rumen. There was a wide individual variation in the production rates of CO 2 between the individual animals. The production rates were not satistically significant between the two groups. (author)

  6. Effects of ratios of non-fibre carbohydrates to rumen degradable ...

    African Journals Online (AJOL)

    p2492989

    digestibility in the MRDP treatment was higher and RDP level equal to 108 (g/kg of DM). ... Keywords: Dairy cows, milk production, non-fibre carbohydrates, rumen .... Van Soest, 1970) with sodium sulphite using heat stable alpha-amylase.

  7. Non-protein nitrogen utilization and microbial synthesis in the rumen

    International Nuclear Information System (INIS)

    Abou Akkada, A.R.; El-Shazly, K.

    1976-01-01

    The distinction between bacterial and protozoal proteins in the rumen has been a difficult task. The use of diaminopimelic acid (DAP) as a marker for bacterial protein and 2-aminoethanephosphonic acid (AEP) for protozoal protein has been examined in the present studies in sheep fed on a semi-purified diet. The microbial protein predicted from DAP and AEP determination was similar to that obtained by the precipitation of proteins. From rates of VFA determination by the in vitro zero-time rate technique, and from measurements of rumen volume using Cr-EDTA colour determination at 550 nm, the total VFA production in 24 h could be calculated, and the ATP generated (mol/day) was calculated using Baldwin and co-workers' transformation value of 2.44. The yield of microbial cells (g/mol ATP) was found to be 28.15, similar to that suggested by Hobson and Summers and Gunzalus and Schuster. Using a sheep double-fistulated in the duodenum proximal to the pyloric sphincter, and blocking the path of the abomasal fluid by inserting an inflated baloon in the second fistula, the abomasal fluid could be collected for 12 h. The total microbial-N as predicted from DAP and AEP was approximately similar to the total non-ammonia-N in the abomasal digesta. It could be concluded that DAP and AEP determination could be used within reasonable limits to differentiate between bacterial and protozoal proteins. A good agreement was found between rates of outflow of the rumen digesta and microbial growth (percentage hourly) as measured by different techniques. The agreement between values of microbial growth rates and rates of outflow of digesta from the rumen with the differences between production and absorption of VFA is an indication of the validity of the techniques employed in the present studies. (author)

  8. The Growth Rate and Efficiency of Rumen Microbial Protein Digestion of Red Clover Silage (Trifolium pratense cv. Sabatron)

    International Nuclear Information System (INIS)

    Asih Kurniawati

    2004-01-01

    (Trifolium pratense cv. Sabatron). Red clover silage supplemented with different level of carbohydrates has been examined using the in-vitro gas production technique. Cumulative gas production, hydro.gen sulfite production, and ammonia was followed and used as indicators of microbial growth rate and extent of protein degradation. Microbial nitrogen production, VFA, and efficiency microbial production was used as indicator of nitrogen use efficiency. 15 N was used as a microbial marker to estimate the amount of nitrogen incorporation into microbial protein. Supplementation of Red clover with increasing 5 levels; 0 g; 0.625 g; 0.15 g; 0.225 g and 0.3 g of maize starch led to graded increase in microbial growth and protein degradation. This was reflected in the increasing gas production and the accumulation of hydrogen sulfite. Diurnal change in ammonia production reflected the microbial utilization of ammonia for protein synthesis. Protein microbe (P<0.001) as VFA (P<0.001) increased due to carbohydrate addition as well as utilization of nitrogen (P<0.001). There was also the efficiency of nitrogen utilization which increased significantly. This result suggested that energy supply can increased efficiency of nitrogen use in the rumen and may reduce nitrogen losses into the environment. (author)

  9. Radiation degradation of silk protein

    International Nuclear Information System (INIS)

    Pewlong, W.; Sudatis, B.; Takeshita, Hidefumi; Yoshii, Fumio; Kume, Tamikazu

    2000-01-01

    Silk fibroin fiber from the domesticated silkworm Bombyx mori was irradiated using an electron beam accelerator to investigate the application of the radiation degradation technique as a means to solubilize fibroin. The irradiation caused a significant degradation of the fiber. The tensile strength of fibroin fiber irradiated up to 2500 kGy decreased rapidly with increasing dose. The presence of oxygen in the irradiation atmosphere enhanced degradation of the tensile strength. The solubilization of irradiated fibroin fiber was evaluated using the following three kinds of solutions: a calcium chloride solution(CaCl 2 /C 2 H 5 OH/H 2 O=1:2:8 in mole ratio), a hydrochloric acid (0.5 N) and a distilled water. Dissolution of fibroin fiber into these solutions was significantly enhanced by irradiation. Especially, an appreciable amount of water soluble proteins was extracted by a distilled water. (author)

  10. Radiation degradation of silk protein

    Energy Technology Data Exchange (ETDEWEB)

    Pewlong, W; Sudatis, B [Office of Atomic Energy for Peace, Bangkok (Thailand); Takeshita, Hidefumi; Yoshii, Fumio; Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2000-03-01

    Silk fibroin fiber from the domesticated silkworm Bombyx mori was irradiated using an electron beam accelerator to investigate the application of the radiation degradation technique as a means to solubilize fibroin. The irradiation caused a significant degradation of the fiber. The tensile strength of fibroin fiber irradiated up to 2500 kGy decreased rapidly with increasing dose. The presence of oxygen in the irradiation atmosphere enhanced degradation of the tensile strength. The solubilization of irradiated fibroin fiber was evaluated using the following three kinds of solutions: a calcium chloride solution(CaCl{sub 2}/C{sub 2}H{sub 5}OH/H{sub 2}O=1:2:8 in mole ratio), a hydrochloric acid (0.5 N) and a distilled water. Dissolution of fibroin fiber into these solutions was significantly enhanced by irradiation. Especially, an appreciable amount of water soluble proteins was extracted by a distilled water. (author)

  11. 6种肉牛常用粗饲料瘤胃降解特性和瘤胃非降解蛋白质的小肠消化率%Ruminal Degradation Characteristics and Small Intestinal Digestibility of Rumen Undegraded Protein of Six Kinds of Commonly Used Roughages for Steers

    Institute of Scientific and Technical Information of China (English)

    陈艳; 张晓明; 王之盛; 彭全辉; 邹华围

    2014-01-01

    本试验旨在研究黑麦草、牛鞭草、甘薯蔓、玉米秸青贮、玉米秸秆和稻草共6种肉牛常用粗饲料的瘤胃降解特性和瘤胃非降解蛋白质( RUP)的小肠消化率。选用3头装有永久性瘤胃瘘管的宣汉阉公牛为试验动物,采用尼龙袋技术评定6种粗饲料的干物质( DM)、粗蛋白质( CP)、中性洗涤纤维( NDF)和酸性洗涤纤维( ADF)瘤胃降解率,并用改进三步法评价RUP的小肠消化率。结果表明:1)黑麦草和甘薯蔓DM有效降解率较高,并与依次降低的牛鞭草、玉米秸青贮、玉米秸秆和稻草差异显著( P<0.05)。CP有效降解率和 ADF有效降解率以黑麦草最高,且与依次降低的甘薯蔓、牛鞭草、玉米秸青贮、玉米秸秆和稻草差异显著( P<0.05)。NDF有效降解率为黑麦草>甘薯蔓>玉米秸青贮>牛鞭草>稻草>玉米秸秆,各粗饲料间差异显著(P<0.05)。2)牛鞭草、玉米秸秆和稻草RUP的小肠消化率差异不显著( P>0.05),并显著高于依次降低的玉米秸青贮、黑麦草和甘薯蔓( P<0.05)。小肠可消化粗蛋白质含量为黑麦草>甘薯蔓>牛鞭草>玉米秸青贮>玉米秸秆>稻草,各粗饲料间差异显著( P<0.05)。由此可见,不同粗饲料瘤胃降解特性不同,为小肠提供可消化粗蛋白质的潜力也不同。黑麦草的 DM、CP、NDF 和ADF在瘤胃的有效降解率最高,牛鞭草、玉米秸秆和稻草RUP的小肠消化率较高,黑麦草和甘薯蔓小肠可消化粗蛋白质含量较高。%This experiment was conducted to determine the ruminal degradation characteristics and small intesti-nal digestibility of rumen undegraded protein( RUP)of 6 kinds of commonly used roughages:ryegrass,hem-arthria compressa,sweet potato stem,corn stover silage,corn stover and rice straw. Three Xuanhan steers fit-ted with permanent ruminal cannulas were used. Nylon

  12. SEVILLA and U.M. LUSTRIA. 2006. Changes in rumen ecosystem and feed dry matter degradability of buffalo which received rumen content of cattle through cross inoculation

    OpenAIRE

    Dicky Pamungkas; Cesar C Sevilla; Ulysses M Lustria

    2006-01-01

    The research was done to identify changes in rumen ecosystem of buffalo which received rumen content of cattle. As much as three head of fistulated male buffaloes (live weight of 450-550 kg) and three fistulated female cattle (live weight 250-380 kg) were used. This experiment was done three stage as follows: pre-inoculation, inoculation and post-inoculation. In Pre-inoculation, the sample of rumen content was taken two hours before morning feeding and directly observed for pH rumen liquor, a...

  13. Measurement of rumen dry matter and neutral detergent fiber degradability of feeds by Fourier-transform infrared spectroscopy

    DEFF Research Database (Denmark)

    Belanche, A.; Weisbjerg, Martin Riis; Allison, G.G.

    2014-01-01

    This study explored the potential of partial least squares (PLS) and Fourier-transform infrared spectroscopy (FTIR) to predict rumen dry matter (DM) and neutral detergent fiber (NDF) degradation parameters of a wide range of feeds for ruminants, as an alternative to the in situ method. In total...... components, such as cellulose, pectin, lignin, cutin, and suberin, but also with nonstructural carbohydrates and certain active compounds. In conclusion, FTIR spectroscopy could be considered a low-cost alternative to in situ measurements in feed evaluation....

  14. Effect of protein degradability on milk production of dairy ewes.

    Science.gov (United States)

    Mikolayunas-Sandrock, C; Armentano, L E; Thomas, D L; Berger, Y M

    2009-09-01

    The objective of this experiment was to determine the effect of protein degradability of dairy sheep diets on milk yield and protein utilization across 2 levels of milk production. Three diets were formulated to provide similar energy concentrations and varying concentrations of rumen-degradable protein (RDP) and rumen-undegradable protein (RUP): 12% RDP and 4% RUP (12-4) included basal levels of RDP and RUP, 12% RDP and 6% RUP (12-6) included additional RUP, and 14% RDP and 4% RUP (14-4) included additional RDP. Diets were composed of alfalfa-timothy cubes, whole and ground corn, whole oats, dehulled soybean meal, and expeller soybean meal (SoyPlus, West Central, Ralston, IA). Estimates of RDP and RUP were based on the Small Ruminant Nutrition System model (2008) and feed and orts were analyzed for Cornell N fractions. Eighteen multiparous dairy ewes in midlactation were divided by milk yield (low and high) into 2 blocks of 9 ewes each and were randomly assigned within block (low and high) to 3 pens of 3 ewes each. Dietary treatments were arranged in a 3 x 3 Latin square within each block and applied to pens for 14-d periods. We hypothesized that pens consuming high-RUP diets (12-6) would produce more milk and milk protein than the basal diet (12-4) and pens consuming high-RDP diets (14-4) would not produce more milk than the basal diet (12-4). Ewes in the high-milk-yield square consumed more dry matter and produced more milk, milk fat, and milk protein than ewes in the low-milk-yield square. There was no effect of dietary treatment on dry matter intake. Across both levels of milk production, the 12-6 diet increased milk yield by 14%, increased milk fat yield by 14%, and increased milk protein yield by 13% compared with the 14-4 and 12-4 diets. Gross N efficiency (milk protein N/intake protein N) was 11 and 15% greater in the 12-6 and 12-4 diets, respectively, compared with the 14-4 diet. Milk urea N concentration was greater in the 12-6 diet and tended to be

  15. Rumen modulatory effect of thyme, clove and peppermint oils in vitro using buffalo rumen liquor.

    Science.gov (United States)

    Roy, Debashis; Tomar, S K; Kumar, Vinod

    2015-02-01

    The present study was conducted to examine the rumen modulatory effect of thyme, clove and peppermint oils on rumen fermentation pattern in vitro using roughage based diet. Thyme, clove and peppermint oils were tested at concentration of 0, 30, 300 and 600 mg/l (ppm) of total culture fluid using in vitro gas production technique in wheat straw based diet (concentrate: Wheat straw 50:50). Different in vitro parameters e.g., total gas production, methane production, nutrient degradability, volatile fatty acid (VFA) production and ammonia nitrogen concentration were studied using buffalo rumen liquor. Thyme oil at higher dose level (600 ppm) reduced (p0.05) in 300 and 600 ppm dose levels. 600 ppm dose level of clove oil reduced (pclove and peppermint oil. Right combination of these essential oils may prove to enhance performance of animals by reducing methane production and inhibiting protein degradation in rumen.

  16. Ruminal and intestinal protein degradability of various seaweed species measured in situ in dairy cows

    DEFF Research Database (Denmark)

    Tayyab, Usama; Novoa-Garrido, Margarita; Roleda, Michael Y.

    2016-01-01

    The use of seaweeds in animal diets is not new. However, little is known about the feed value of seaweed, both in terms of chemical composition and protein digestibility, and regarding variation between species and season. In this study, eight seaweed species of the genus Acrosiphonia, Alaria......, Laminaria, Mastocarpus, Palmaria, Pelvetia, Porphyra, and Ulva were sampled in spring (March) and autumn (October and November) 2014 at the coast of Bodø in Northern Norway, and were analysed for chemical composition, in situ rumen degradability and total tract crude protein (CP) digestibility. Ash content...... for Pelvetia (90 g/kg DM). Spring samples were higher in CP than autumn samples. The effective degradability estimated at 5% rumen passage rate (ED5) of CP varied between species (P Ulva (240 g...

  17. The Role of Ciliate Protozoa in the Rumen.

    Science.gov (United States)

    Newbold, Charles J; de la Fuente, Gabriel; Belanche, Alejandro; Ramos-Morales, Eva; McEwan, Neil R

    2015-01-01

    First described in 1843, Rumen protozoa with their striking appearance were assumed to be important for the welfare of their host. However, despite contributing up to 50% of the bio-mass in the rumen, the role of protozoa in rumen microbial ecosystem remains unclear. Phylogenetic analysis of 18S rDNA libraries generated from the rumen of cattle, sheep, and goats has revealed an unexpected diversity of ciliated protozoa although variation in gene copy number between species makes it difficult to obtain absolute quantification. Despite repeated attempts it has proven impossible to maintain rumen protozoa in axenic culture. Thus it has been difficult to establish conclusively a role of ciliate protozoa in rumen fiber degradation. The development of techniques to clone and express ciliate genes in λ phage, together with bioinformatic indices to confirm the ciliate origin of the genes has allowed the isolation and characterization of fibrolytic genes from rumen protozoa. Elimination of the ciliate protozoa increases microbial protein supply by up to 30% and reduces methane production by up to 11%. Our recent findings suggest that holotrich protozoa play a disproportionate role in supporting methanogenesis whilst the small Entodinium are responsible for much of the bacterial protein turnover. As yet no method to control protozoa in the rumen that is safe and practically applicable has been developed, however a range of plant extract capable of controlling if not completely eliminating rumen protozoa have been described.

  18. Effect of feeding complete feed block containing rumen protected protein, non-protein nitrogen and rumen protected fat on improving body condition and carcass traits of cull ewes.

    Science.gov (United States)

    Bhatt, R S; Sahoo, A

    2017-12-01

    Nutrient utilization, body condition and carcass traits of cull ewes were studied in three dietary regimens based on complete feed block (CFB) feeding to control (C) with rumen protected protein (RPP), CU [RPP + urea (6 g/kg)] and CUF [RPP + urea + rumen protected fat (RPF; 40 g/kg)]. The RPP component (g/kg) in C had 1% formaldehyde-treated soy flakes 50, mustard cake 50 and sesame cake 30. The mustard and sesame cakes were replaced with urea on equivalent N basis in CU and CUF. The ewes were offered ad libitum CFB composed (g/kg) of concentrate 650, roughage 300 and molasses 50. The digestibility of OM and EE was higher (p Ewes in all the groups showed an improvement in carcass traits at 90 day. The pre-slaughter weight was higher (p ewes. Journal of Animal Physiology and Animal Nutrition © 2017 Blackwell Verlag GmbH.

  19. Production indicators and health status of calves fed different amounts of rumen undegradable starch and protein

    Directory of Open Access Journals (Sweden)

    Tomislav Koturić

    2017-01-01

    Full Text Available The aim of this study was to determine whether increase in proportion of the rumen undegradable starch (RUS and rumen undegradable protein (RUP affects the production performance and health status of calves. The experiment was done on 36 Holstein, seven-day-old calves, divided into three groups of 12 calves, with equal sex ratio. The experiment was conducted in two periods. In the first period, calves were fed with full pasteurized milk and milk replacer and additionally fed with starter mixture with different proportions of rumen undegradable protein and starch: Group I 36.6% RUP and 16.5% RUS, Group II 49.1% RUP and 27.6% RUS and Group III 53.5% RUP and 36.5% RUS. In the second period, calves were fed with milk replacer and grower mixture with different proportions of rumen undegradable protein and starch: Group I 33.5% RUP and 15.8% RUS, Group II 48% RUP and 26.3% RUS and Group III 54.3% RUP and 34.6% RUS. In the first period, calves from the Group III had significantly (P<0.01 higher body weight compared to calves in Group I and II (74.75; 59.36; 66.58 kg, as well as daily weight gain (0.76, 0.49, 0.61 kg/d. At the end of the experiment, there was no significant difference in body weight and daily weight gain. The calves in Group I and III had significantly (P<0.05 higher consumption of starter mixture compared to the calves in Group II (7.48; 7.11; 4.33 kg/d, and a significantly (P<0.05 higher overall feed consumption compared to the calves in Group II. The calves in Group II and III had significantly (P<0.05 better feed conversion ratio than the calves in Group I (1.37; 1.50; 2.08 kg/kg. The results of health monitoring (diarrhea, pneumonia indicate a different proportion of rumen undegradable starch and protein ratio did not have significant effect on calves’ health.

  20. Radiation degradation of silk protein

    Energy Technology Data Exchange (ETDEWEB)

    Wachiraporn Pewlong; Boonya Sudatis [Office of Atomic Energy for Peace, Bangkok (Thailand); Takeshita, Hidefumi; Yoshii, Fumio; Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2000-09-01

    Silk fibroin fiber from the domesticated silkworm Bombyx mori was irradiated in the dose range up to 2500 kGy using an electron beam accelerator to apply the radiation degradation technique as a means to solubilize fibroin. The tensile strength of irradiated fibroin fiber decreased with increasing dose and the presence of oxygen in the irradiation atmosphere enhanced the degradation. The solubilization of irradiated fibroin fiber was evaluated using the following three kinds of solutions: calcium chloride solution (CaCl{sub 2}/C{sub 2}H{sub 5}OH/H{sub 2}O = 1 : 2 : 8 in mole ratio), hydrochloric acid (0.5N) and distilled water. Dissolution of fibroin fiber into these solutions was significantly enhanced by irradiation. Especially, an appreciable amount of water-soluble protein was extracted by distilled water. (author)

  1. Determination of possible effects of mineral concentration on protein synthesis by rumen microbes in vitro

    International Nuclear Information System (INIS)

    Nikolic, J.A.; Jovanovic, M.; Andric, R.

    1976-01-01

    The aim of the present investigation was to determine the effect of different concentrations of sulphide, magnesium and zinc on protein synthesis by rumen micro-organisms in vitro. Rumen content was taken from a young bull fed a diet based on maize and dried sugar beet pulp (2/1) supplemented with urea. The rate of incorporation of 35 S from Na 2 35 SO 4 in relation to the mean specific radioactivity of the sulphide pool was used to estimate the overall rate of microbial protein synthesis. It was found that the rate of protein synthesis and the net rate of utilization of ammonia-N were not affected by differences in mean sulphide concentration from 3.6-8.0 mg/litre. The rate of reduction of sulphate appeared not to be affected by the addition of sodium sulphide to the medium. The rate and efficiency of protein synthesis by rumen micro-organisms were not significantly affected by increasing the concentration of total magnesium from 8.4-15.3 mg/100 ml. The values for soluble magnesium varied widely (1.2-7.8 mg/100 ml), and appeared to be partly dependent on the pH of the medium. Zinc concentrations varying from 5.2-12.4 mg/litre did not influence the overall rate of protein synthesis, although the efficiency tended to be higher when the concentration of zinc was greater. Concentrations of soluble zinc were low (0.3-1.15 mg/litre), and not influenced by changes in the concentration of total zinc. It was concluded that increasing the concentrations of the examined elements above the basic values did not lead consistently to an improved production of microbial protein but, on the other hand, had no obvious detrimental effect on microbial metabolic activity within the limits studied. (author)

  2. Effect of rumen-undegradable protein supplementation and fresh forage composition on nitrogen utilization of dairy ewes.

    Science.gov (United States)

    Mikolayunas, C; Thomas, D L; Armentano, L E; Berger, Y M

    2011-01-01

    Previous trials with dairy ewes fed stored feeds indicate a positive effect of rumen-undegradable protein (RUP) supplementation on milk yield. However, dairy sheep production in the United States is primarily based on grazing mixed grass-legume pastures, which contain a high proportion of rumen-degradable protein. Two trials were conducted to evaluate the effects of high-RUP protein supplementation and fresh forage composition on milk yield and N utilization of lactating dairy ewes fed in confinement or on pasture. In a cut-and-carry trial, 16 multiparous dairy ewes in mid-lactation were randomly assigned to 8 pens of 2 ewes each. Pens were randomly assigned 1 of 2 protein supplementation treatments, receiving either 0.0 or 0.3 kg of a high-RUP protein supplement (Soy Pass, LignoTech USA Inc., Rothschild, WI) per day. Within supplementation treatment, pens were randomly assigned to 1 of 4 forage treatments, which were applied in a 4×4 Latin square design for 10-d periods. Forage treatments included the following percentages of orchardgrass:alfalfa dry matter: 25:75, 50:50, 75:25, and 100:0. No interactions were observed between supplement and forage treatments. Supplementation with a high-RUP source tended to increase milk yield by 9%. Milk yield, milk protein yield, milk urea N, and urinary urea N excretion increased linearly with increased percentage of alfalfa. Milk N efficiency was greatest on the 100% orchardgrass diet. In a grazing trial, 12 multiparous dairy ewes in mid lactation were randomly assigned to 3 groups of 4 ewes each. Within group, 2 ewes were randomly assigned to receive either 0.0 or 0.3 kg of a high-RUP protein supplement (SoyPlus, West Central Cooperative, Ralston, IA) per day. Grazing treatments were arranged in a 3×3 Latin square design and applied to groups for 10-d periods. Ewes grazed paddocks that contained the following percentages of surface area of pure stands of orchardgrass:alfalfa: 50:50, 75:25, and 100:0. No interactions were

  3. Sporulation capability and amylosome conservation among diverse human colonic and rumen isolates of the keystone starch‐degrader Ruminococcus bromii

    Science.gov (United States)

    Mukhopadhya, Indrani; Moraïs, Sarah; Laverde‐Gomez, Jenny; Sheridan, Paul O.; Walker, Alan W.; Kelly, William; Klieve, Athol V.; Ouwerkerk, Diane; Duncan, Sylvia H.; Louis, Petra; Koropatkin, Nicole; Cockburn, Darrell; Kibler, Ryan; Cooper, Philip J.; Sandoval, Carlos; Crost, Emmanuelle; Juge, Nathalie; Bayer, Edward A.

    2017-01-01

    Summary Ruminococcus bromii is a dominant member of the human colonic microbiota that plays a ‘keystone’ role in degrading dietary resistant starch. Recent evidence from one strain has uncovered a unique cell surface ‘amylosome’ complex that organizes starch‐degrading enzymes. New genome analysis presented here reveals further features of this complex and shows remarkable conservation of amylosome components between human colonic strains from three different continents and a R. bromii strain from the rumen of Australian cattle. These R. bromii strains encode a narrow spectrum of carbohydrate active enzymes (CAZymes) that reflect extreme specialization in starch utilization. Starch hydrolysis products are taken up mainly as oligosaccharides, with only one strain able to grow on glucose. The human strains, but not the rumen strain, also possess transporters that allow growth on galactose and fructose. R. bromii strains possess a full complement of sporulation and spore germination genes and we demonstrate the ability to form spores that survive exposure to air. Spore formation is likely to be a critical factor in the ecology of this nutritionally highly specialized bacterium, which was previously regarded as ‘non‐sporing’, helping to explain its widespread occurrence in the gut microbiota through the ability to transmit between hosts. PMID:29159997

  4. Selective ligninolysis of wheat straw and wood chips by the white-rot fungus Lentinula edodes and its influence on in vitro rumen degradability

    OpenAIRE

    Kuijk, van, S.J.A.; Rio, del, José C.; Rencoret, Jorge; Gutiérrez, Ana; Sonnenberg, A.S.M.; Baars, J.J.P.; Hendriks, W.H.; Cone, J.W.

    2016-01-01

    Background: The present work investigated the influence of lignin content and composition in the fungaltreatment of lignocellulosic biomass in order to improve rumen degradability. Wheat straw and wood chips,differing in lignin composition, were treated with Lentinula edodes for 0, 2, 4, 8 and 12 wk and the changesoccurring during fungal degradation were analyzed using pyrolysis-gas chromatography-mass spectrometryand detergent fiber analysis.Results: L. edodes preferentially degraded lignin,...

  5. The effect of age on in sacco estimates of rumen dry matter and ...

    African Journals Online (AJOL)

    This study was conducted to determine whether rumen dry matter and crude protein degradability in calves aged 8-10 weeks differs from that in mature cows. Five Holstein bull calves were rumen-fistulated at six weeks of age and were used in consecutive weekly 24 h trials from 8-20 weeks of age. Dry matter and crude ...

  6. Effects of alternative protein sources on rumen microbes and productivity of dairy cows

    Directory of Open Access Journals (Sweden)

    Metha Wanapat

    2011-01-01

    Full Text Available This experiment was conducted to investigate the effect of various protein sources on digestibility, rumen fermentation, milk yield and milk composition in dairy cows. Four Holstein Friesian native crossbred cows in early lactating were randomly assigned according to a 4x4 Latin square design. The dietary treatments containing different protein sources in concentrate diets were soybean meal (SBM, cassava hay (CH, Leucaena leucocephala (LL and yeast-fermented cassava chips (YEFECAP, with ad libitum intake of urea-treated rice straw. Digestibility of DM, OM, NDF and ADF was not different among treatments (P>0.05 while CP digestibility was highest (P<0.05 in CH and YEFECAP supplemented groups. Ruminal NH3-N and BUN concentrations varied among protein sources and were highest in SBM and LL fed groups (P<0.05. Ruminal total volatile fatty acid (VFA and propionic acid were found highest in cows receiving CH and YEFECAP (P<0.05. Ruminal fungi, proteolytic and cellulolytic bacteria were highest when YEFECAP was supplemented. Milk fat and milk protein were significantly increased (P<0.05 in cows fed with CH and YEFECAP. Based on this study, it was concluded that providing CH or YEFECAP as protein source in concentrate diets could improve rumen fermentation and milk production in lactating dairy cows fed on rice straw.

  7. The contribution of mathematical modeling to understanding the dynamic aspects of rumen metabolism

    Directory of Open Access Journals (Sweden)

    André Bannink

    2016-11-01

    Full Text Available All rumen models cover the main drivers of variation in rumen function, which are feed intake, the differences between feedstuffs and feeds in their intrinsic rumen degradation characteristics, and fractional outflow rate of fluid and particulate matter. Dynamic modeling approaches are best suited to the prediction of more nuanced responses in rumen metabolism, and represent the dynamics of the interaction between substrates and micro-organisms and inter-microbial interactions. The concepts of dynamics are discussed for the case of rumen starch digestion as influenced by starch intake rate and frequency of feed intake, and for the case of fermentation of fiber in the large intestine. Adding representations of new functional classes of micro-organisms (i.e. with new characteristics from the perspective of whole rumen function in rumen models only delivers new insights if complemented by the dynamics of their interactions with other functional classes. Rumen fermentation conditions have to be represented due to their profound impact on the dynamics of substrate degradation and microbial metabolism. Although the importance of rumen acidity is generally acknowledged, more emphasis is needed on predicting its variation as well as variation in the processes that underlie rumen fluid dynamics. The rumen wall has an important role in adapting to rapid changes in the rumen environment, clearing of volatile fatty acids (VFA, and maintaining rumen pH within limits. Dynamics of rumen wall epithelia and its role in VFA absorption needs to be better represented in models which aim to predict rumen responses across nutritional or physiological states. For a detailed prediction of rumen N balance there is merit in a dynamic modeling approach compared to the static approaches adopted in current protein evaluation systems. Improvement is needed on previous attempts to predict rumen VFA profiles, and this should be pursued by introducing factors that relate more

  8. Ruminal and intestinal protein degradability of various seaweed species measured in situ in dairy cows

    DEFF Research Database (Denmark)

    Tayyab, Usama; Novoa-Garrido, Margarita; Roleda, Michael Y.

    2016-01-01

    , Laminaria, Mastocarpus, Palmaria, Pelvetia, Porphyra, and Ulva were sampled in spring (March) and autumn (October and November) 2014 at the coast of Bodø in Northern Norway, and were analysed for chemical composition, in situ rumen degradability and total tract crude protein (CP) digestibility. Ash content......The use of seaweeds in animal diets is not new. However, little is known about the feed value of seaweed, both in terms of chemical composition and protein digestibility, and regarding variation between species and season. In this study, eight seaweed species of the genus Acrosiphonia, Alaria....../kg CP). Digestible rumen escape protein (DEP) varied significantly between species (P Laminaria, Mastocarpus and Palmaria can supply...

  9. Rumen degradation characteristics of ryegrass herbage and ryegrass silage are affected by interactions between stage of maturity and nitrogen fertilisation rate

    NARCIS (Netherlands)

    Heeren, J.A.H.; Podesta, S.C.; Hatew, B.; Klop, G.; Laar, van H.; Bannink, A.; Warner, D.; Jonge, de L.H.; Dijkstra, J.

    2014-01-01

    The objective of this experiment was to evaluate interaction effects between stage of maturity and N fertilization rate on rumen degradation characteristics determined with nylon bag incubations of ryegrass herbages and ryegrass silage. Grass herbage (n = 4) was cut after 3 or 5 weeks of regrowth

  10. Selective ligninolysis of wheat straw and wood chips by the white-rot fungus Lentinula edodes and its influence on in vitro rumen degradability

    NARCIS (Netherlands)

    Kuijk, van S.J.A.; Rio, del José C.; Rencoret, Jorge; Gutiérrez, Ana; Sonnenberg, A.S.M.; Baars, J.J.P.; Hendriks, W.H.; Cone, J.W.

    2016-01-01

    Background: The present work investigated the influence of lignin content and composition in the fungal
    treatment of lignocellulosic biomass in order to improve rumen degradability. Wheat straw and wood chips,
    differing in lignin composition, were treated with Lentinula edodes for 0, 2, 4, 8

  11. Development of Equation Based on Urinary Purine Derivatives to Estimate Rumen Microbial Protein Production in Goats

    International Nuclear Information System (INIS)

    Jetana, Thongsuk; Abdullah, Norhani; Liang, Boo Juan; Syed Salim, Syed Jalaludin; Ho, Wan Yin

    2003-06-01

    Three experiments were conducted at the farm of the Universiti Putra Malaysia, Serdang, Selangor, Malaysia, to establish a model as an index for estimating rumen microbial protein production. In Experiment 1, six Ferral male goats (wt. 40.2±4.6 kg) were used to determine the endogenous purine derivatives (PD) excreted in the urine by fasting. In Experiment 2, four Ferral male goats (wt. 39.6±1.8 kg) were used to measure the proportion of plasma PD excreted in the urine by using [ 14 C]-uric acid as a marker at two levels of feed intake (40% and 80% voluntary intake), using an incomplete 2x4 Latin square experimental design. The feed consisted of 40% oil palm frond and 60% concentrate (OPFC). In Experiment 3, four Ferral male goats fed (OPFC)) were slaughtered and rumen contents were taken for measurements of purine and total nitrogen contents of mixed rumen microbes. The results showed that endogenous PD (allantoin, uric acid, xanthine and hypoxanthine) excreted in the urine obtained by the fasting trial was 202±17 μmol/kg BW 0 . 75 d - 1. The average percentage recovery of plasma PD excretion in the urine by using [ 14 C)-uric acid as a marker was 83±2.0% (cv=6.88, ranged 76.3-91.4%, n=8). Percentage recovery was not affected by levels of feed intake. The ratio of purine N: total N in the mixed rumen liquid associated bacteria (LAB) was 0.085. In this study, a preliminary model for goats was established by using the information from the recovery of labeled PD [ 14 C]-uric acid and the fasting PD excretion. The model obtained was Y 0.83X + 0.202 x BW 0 . 75 , where Y = PD excretion in the urine (mmol/d) X PD absorption at small intestine (mmol/d) BW 0 . 75 = Metabolic body weight (kg) Thus the microbial nitrogen based on total PD (MNpd) can be calculated as follows: MNpd = 70 x X = 0.992 x X (g/d) 0.085 x 0.83 x 1000 where 0.085 is the ratio of purine-N: total N in mixed rumen microbes, 0.83 is the average of digestibility of microbial purine from published

  12. Effect of dietary protein levels on rumen metabolism and milk yield in mid-lactating cows under hot and humid conditions

    NARCIS (Netherlands)

    Thiangtum, W.; Schonewille, J.T.; Yawongsa, A.; Rukkwamsuk, T.; Kanjanapruthipong, J.; Verstegen, M.W.A.; Hendriks, W.H.

    2014-01-01

    An experiment was conducted to investigate the effects of 2 levels of dietary Crude Protein (CP) in concentrates with similar proportions of Rumen Undegradable Protein (RUP) on rumen metabolism, milk yield and composition in mid lactating cows in Thailand. Eight 87.5% Holsteinx12.5% indigenous

  13. Effect of dietary protein levels on rumen metabolism and milk yield in mid-lactating cows under hot and humid conditions.

    NARCIS (Netherlands)

    Thiangtum, W; Schonewille, Thomas; Yawongsa, A; Rukkwamsuk, T; Kanjanapruthipon, J; Verstegen, M.W.A.; Hendriks, Wouter

    2014-01-01

    An experiment was conducted to investigate the effects of 2 levels of dietary Crude Protein (CP) in concentrates with similar proportions of Rumen Undegradable Protein (RUP) on rumen metabolism, milk yield and composition in mid lactating cows in Thailand. Eight 87.5% Holsteinx12.5% indigenous

  14. Functional analyses of multiple lichenin-degrading enzymes from the rumen bacterium Ruminococcus albus 8.

    Science.gov (United States)

    Iakiviak, Michael; Mackie, Roderick I; Cann, Isaac K O

    2011-11-01

    Ruminococcus albus 8 is a fibrolytic ruminal bacterium capable of utilization of various plant cell wall polysaccharides. A bioinformatic analysis of a partial genome sequence of R. albus revealed several putative enzymes likely to hydrolyze glucans, including lichenin, a mixed-linkage polysaccharide of glucose linked together in β-1,3 and β-1,4 glycosidic bonds. In the present study, we demonstrate the capacity of four glycoside hydrolases (GHs), derived from R. albus, to hydrolyze lichenin. Two of the genes encoded GH family 5 enzymes (Ra0453 and Ra2830), one gene encoded a GH family 16 enzyme (Ra0505), and the last gene encoded a GH family 3 enzyme (Ra1595). Each gene was expressed in Escherichia coli, and the recombinant protein was purified to near homogeneity. Upon screening on a wide range of substrates, Ra0453, Ra2830, and Ra0505 displayed different hydrolytic properties, as they released unique product profiles. The Ra1595 protein, predicted to function as a β-glucosidase, preferred cleavage of a nonreducing end glucose when linked by a β-1,3 glycosidic bond to the next glucose residue. The major product of Ra0505 hydrolysis of lichenin was predicted to be a glucotriose that was degraded only by Ra0453 to glucose and cellobiose. Most importantly, the four enzymes functioned synergistically to hydrolyze lichenin to glucose, cellobiose, and cellotriose. This lichenin-degrading enzyme mix should be of utility as an additive to feeds administered to monogastric animals, especially those high in fiber.

  15. In vitro estimation of rumen microbial protein synthesis of water buffaloes using 30S as tracer

    International Nuclear Information System (INIS)

    Hendratno, C.; Abidin, Z.; Bahaudin, R.; Sastrapradja, D.

    1977-01-01

    An experiment to study the effect of diet and individual differences of animals on the in vitro estimation of rumen microbial protein synthesis in young female water buffaloes using the technique of inorganic 35 S incorporation, is described. The dietary treatments were four combinations of roughage supplemented with cassava meal. From the value of rate constant for dilution of radioactivity in the sulphide pool and percentage of inorganic 35 S incorporated into microbial protein, it can be concluded that individual differences of animals have no influence on the efficiency of microbial protein synthesis. Feed composition, on the other hand, tends to have some influence on the efficiency of protein synthesis(P3O.15). (author)

  16. Evaluation of sonication treatment and buffer composition on rumen bacteria protein extraction and carboxymethylcellulase activity.

    Science.gov (United States)

    Prauchner, Carlos A; Kozloski, Gilberto V; Farenzena, Roberta

    2013-05-01

    The methodological procedures for studying the fibrolytic activity of rumen bacteria are not clearly established. In this study the efficiency of sonication treatment and buffer composition (i.e. buffer varying in tonicity or pH) on the level of protein extraction from the residue of forage samples incubated in the rumen of a grazing steer and the effect of buffer composition or CaCl₂ concentration on the carboxymethylcellulase (CMCase) activity of the released protein were evaluated. The amount of protein released from the residue of incubation was higher (P buffer pH (P buffer. Both linear and quadratic effects (P buffer pH on CMCase activity were significant, with CMCase activity being maximal at pH 5.4-6.1. CMCase activity was higher (P buffer solution that includes a calcium source. When pH is not a treatment factor, the buffer pH should be between 5.5 and 6. © 2012 Society of Chemical Industry.

  17. Effect of experimentally increased protein supply to postpartum dairy cows on plasma protein synthesis, rumen tissue proliferation, and immune homeostasis.

    Science.gov (United States)

    Larsen, M; Røntved, C M; Theil, P K; Khatun, M; Lauridsen, C; Kristensen, N B

    2017-05-01

    The effect of experimentally increasing the postpartum protein supply on plasma protein synthesis, rumen tissue proliferation, and immune homeostasis was studied using 8 periparturient Holstein cows in a complete randomized design. At calving, cows were assigned to abomasal infusion of water (CTRL) or casein (CAS) in addition to a lactation diet. Casein infusion was gradually decreased from 696 ± 1 g/d at +2 d relative to calving (DRTC) to 212 ± 10 g/d at +29 DRTC to avoid excessive supply. Synthesis rate of plasma proteins was measured at -14, +4, +15, and +29 DRTC by measuring [C]Phe isotopic enrichment in arterial plasma free Phe, total plasma proteins, and albumin after 3, 5, and 7 h of jugular ring[C]Phe infusion. Plasma volume was determined at +4 and +29 DRTC by dilution of a [I]BSA dose. Synthesis rate of tissue protein in biopsied rumen papillae was determined by measuring [C]Phe isotopic enrichment, and mRNA expression of selected genes was measured by real-time qPCR. Total and differential leukocyte counts were performed and immune responsiveness of monocytes was evaluated by tumor necrosis factor ɑ (TNFɑ) concentration on ex vivo whole blood stimulation with Escherichia coli lipopolysaccharide (LPS) and responsiveness of T-lymphocytes by interferon γ (IFNγ) concentration on stimulation with Staphylococcus aureus enterotoxin β (SEB). Further, ELISA plasma concentrations of IgM, IgA, and IgG were determined. The DRTC affected the majority of investigated parameters as expected. The CAS treatment increased milk protein yield (P = 0.04), and tended to lower TNFɑ (P = 0.06), and lowered IFNγ (P = 0.03) responsiveness per monocyte and lymphocyte, respectively, compared with CTRL. Further, fractional synthesis rate of albumin was greater at +4 DRTC for CAS compared with CTRL but did not differ by +29 DRTC (interaction: P = 0.01). In rumen papillae, synthesis rate of tissue protein was greater for CAS compared with CTRL (P protein supply seem to

  18. Estudo dos protozoários ciliados em bovinos consumindo dietas com diferentes níveis de proteína não degradável no rúmen 1 - DOI: 10.4025/actascianimsci.v25i1.2146 Ciliated protozoa in the rumen of cattle under different levels of undegradable protein diets - DOI: 10.4025/actascianimsci.v25i1.2146

    Directory of Open Access Journals (Sweden)

    José Aparecido Cunha

    2003-04-01

    Full Text Available O experimento foi realizado no Departamento de Zootecnia da Faculdade de Zootecnia e Engenharia de Alimentos da Universidade de São Paulo, no Campus da USP, em Pirassununga, Estado de São Paulo, e teve por objetivos estudar a variação de pH e identificar e quantificar os protozoários ciliados do rúmen de bovinos submetidos a dietas com diferentes níveis de proteína não degradável no rúmen (PNDR, denominados tratamentos A-24%, B-30%, C-36% e D-42%. Todas as rações foram formuladas para serem isoproteicas e isoenergéticas. A dieta teve um teor de PB de 11% e 63% de NDT, e os ingredientes que a compunham foram: farelo de soja, milho (grãos, aveia (grãos, sorgo (grãos, silagem de milho, farinha de sangue, farinha de carne, calcário, sal e uréia, os quais foram oferecidas a quatro bovinos adultos, mestiços europeu x zebu, dotados de cânulas ruminais, com pesos médios de 600kg. Todos os animais eram castrados. O delineamento estatístico utilizado foi o de Quadrado Latino (4x4. O ensaio experimental foi subdividido em 4 períodos, cada um com 21 dias de adaptação, e, nos dias 23 e 24 de cada período, foram feitas as colheitas das amostras do líquido ruminal antes da alimentação e de 2 em 2 horas, durante um período de 8 horas, seguidas imediatamente da determinação do pH. Os resultados evidenciaram que o pH foi influenciado pelos níveis de PNDR, revelando diferença significativa no efeito tempo (horas após a alimentação, (p Entodinium spp (p The experiment was carried out at the Zootechny and Food Engineering Faculty of Universidade de São Paulo (São Paulo University - Brazil, and it aimed to study the pH variation and also to identify and to qualify the ciliated protozoa in the rumen of cattle under 24%, 30%, 36% and 42% of undegradable protein (PNDR diets, named as treatments A, B, C and D, respectively. All the diets had the same protein and energy concentration (11% CP and 63% TDN, and they were composed of

  19. Feed degradability, rumen fermentation and blood metabolites in response to essential oil addition to fistulated non-lactating dairy cow diets.

    Science.gov (United States)

    Suksombat, Wisitiporn; Nanon, Atitthan; Meeprom, Chayapol; Lounglawan, Pipat

    2017-09-01

    The effects of essential oils (EOs) on ruminal nutrient disappearance, rumen fermentation and blood metabolites in fistulated non-lactating dairy cows were studied. Four fistulated non-lactaing dairy cows were used in a 4 × 4 Latin square design; the experiment consisted of four periods of 21 days in each period, with the first 14 days for adaptation followed by 7 days of measurement period. Animals were fed 3 kg/day of 21% crude protein (CP) concentrate and ad libitum corn silage. Treatments were: (i) control; (ii) 2 mL Allicin/cow/day; (iii) 2 mL zingiberene/cow/day; and (iv) 2 mL citral/cow/day. The results demonstrated that EOs increased dry matter and neutral detergent fiber degradabilities at 48 and 72 h, but had no effect on acid detergent fiber and CP degradabilities. EOs did not change ruminal pH, ammonia nitrogen, protozoa, volatile fatty acid concentrations and blood glucose but reduced blood urea nitrogen at 4 h. © 2017 Japanese Society of Animal Science.

  20. Intake, Rumen Degradation and Utilisation of Urea-Ammoniated Grass Hay by Kacang Goats as Affected by Supplementation of Sun-dried Fish or Fishmeal

    Directory of Open Access Journals (Sweden)

    MR Weibsjerg

    2012-05-01

    Full Text Available Six mature male Kacang goats were involved in an experiment arranged following a duplicate 3 x 3 Latin Square Design. The objectives of this experiment was to study the effect of supplementation of dried fish as compared to fishmeal on intake, digestion, rumen environment and nitrogen used by the local meat type Kacang goats maintained on urea-treated low quality grass hay. The treatments were G: goats were fed with ad libitum access of urea treated grass hay and 100 g/d putak, SDF: G plus 18.4 g sun dried fish, and FM: G plus 19.7 g/d fishmeal. The supplement in SDF and FM were at equal CP level. Intake of urea treated grass hay tended to increase (P=0.08 with supplementation. Dry matter digestibility particularly that of CP was improved by supplementation. Rumen environment was slightly modified by supplementation. Rumen pH was reduced while ammonia concentration was increased. Rumen degradation of the treated grass hay did not differ when incubated in the rumen of goats with different diets. Nitrogen balance was significantly improved (P<0.05 by fishmeal supplementation. In all parameters measured in this experiment, the incremental effects did not differ between fishmeal type. This indicate that there is no further advantage of preparing fishmeal other than sun-drying in improving the utilisation of low quality urea-treated grass hay.

  1. Effects of experimentally increased protein supply to postpartum dairy cows on plasma protein synthesis, rumen tissue proliferation, and immune homeostasis

    DEFF Research Database (Denmark)

    Larsen, Mogens; Røntved, Christine Maria; Theil, Peter Kappel

    2017-01-01

    The effect of experimentally increasing the postpartum protein supply on plasma protein synthesis, rumen tissue proliferation, and immune homeostasis was studied using 8 periparturient Holstein cows in a complete randomized design. At calving, cows were assigned to abomasal infusion of water (CTRL......) or casein (CAS) in addition to a lactation diet. Casein infusion was gradually decreased from 696 ± 1 g/d at +2 d relative to calving (DRTC) to 212 ± 10 g/d at +29 DRTC to avoid excessive supply. Synthesis rate of plasma proteins was measured at –14, +4, +15, and +29 DRTC by measuring [13C]Phe isotopic...... enrichment in arterial plasma free Phe, total plasma proteins, and albumin after 3, 5, and 7 h of jugular ring[13C]Phe infusion. Plasma volume was determined at +4 and +29 DRTC by dilution of a [125I]BSA dose. Synthesis rate of tissue protein in biopsied rumen papillae was determined by measuring [13C...

  2. Optimal Cultivation Time for Yeast and Lactic Acid Bacteria in Fermented Milk and Effects of Fermented Soybean Meal on Rumen Degradability Using Nylon Bag Technique

    Directory of Open Access Journals (Sweden)

    S. Polyorach

    2016-09-01

    Full Text Available The objectives of this study were to determine an optimal cultivation time for populations of yeast and lactic acid bacteria (LAB co-cultured in fermented milk and effects of soybean meal fermented milk (SBMFM supplementation on rumen degradability in beef cattle using nylon bag technique. The study on an optimal cultivation time for yeast and LAB growth in fermented milk was determined at 0, 4, 8, 24, 48, 72, and 96 h post-cultivation. After fermenting for 4 days, an optimal cultivation time of yeast and LAB in fermented milk was selected and used for making the SBMFM product to study nylon bag technique. Two ruminal fistulated beef cattle (410±10 kg were used to study on the effect of SBMFM supplementation (0%, 3%, and 5% of total concentrate substrate on rumen degradability using in situ method at incubation times of 0, 2, 4, 6, 12, 24, 48, and 72 h according to a Completely randomized design. The results revealed that the highest yeast and LAB population culture in fermented milk was found at 72 h-post cultivation. From in situ study, the soluble fractions at time zero (a, potential degradability (a+b and effective degradability of dry matter (EDDM linearly (p<0.01 increased with the increasing supplemental levels and the highest was in the 5% SBMFM supplemented group. However, there was no effect of SBMFM supplement on insoluble degradability fractions (b and rate of degradation (c. In conclusion, the optimal fermented time for fermented milk with yeast and LAB was at 72 h-post cultivation and supplementation of SBMFM at 5% of total concentrate substrate could improve rumen degradability of beef cattle. However, further research on effect of SBMFM on rumen ecology and production performance in meat and milk should be conducted using in vivo both digestion and feeding trials.

  3. An assessment of differences in the ruminal degradability and intestinal digestibility of crude protein in brewer’s grains and maize draff

    Directory of Open Access Journals (Sweden)

    Vladimír Majer

    2012-01-01

    Full Text Available The submitted thesis aims to assess the differences between the ruminal degradability and intestinal digestibility of crude protein contained in brewer’s grains (BG and maize draff (AMG. The effectiveness of ruminal degradability was tested using the “in sacco” method on 3 dry Holstain cows fitted with rumen cannulas. The dynamics of ruminal degradability of crude protein (CP was detected after 0, 4, 8, 16, and 24 hours of samples incubation in the rumen. The intestinal digestibility of crude protein undegradable in the rumen was determined using the “mobile bag” method on 3 dry Holstain cows fitted with duodenal cannulas. The crude protein degradability of BG was detected in the above-mentioned hours (%: 4.06; 18.16; 32.40; 38.56, and 50.70; crude protein degradability of AMG: 42.04; 63.56; 84.47; 85.16, and 87.19. The effectiveness of rumen degradability of BG crude protein at the rate of passage of rumen content 6 % per hour was calculated at 35.33 % and that of AMG, at 76.29 %. Intestinal digestibility of BG crude protein and dry matter at the rate of passage of intestinal content 6 % per hour was calculated at 79.41 % and 22.84 %, respectively, and that of AMG, at 57.01 % and 11.33 %, respectively. The differences between the indicators of both feedstuffs were significant (P < 0.05. The results show that BG are mostly a source of crude protein with higher intestinal digestibility than AMG.

  4. Effect of Saccharomyces cerevisiae on alfalfa nutrient degradation characteristics and rumen microbial populations of steers fed diets with different concentrate-to-forage ratios.

    Science.gov (United States)

    Ding, Gengzhi; Chang, Ying; Zhao, Liping; Zhou, Zhenming; Ren, Liping; Meng, Qingxiang

    2014-01-01

    Live yeast (Saccharomyces cerevisiae) constitutes an effective additive for animal production; its probiotic effect may be related to the concentrate-to-forage ratio (CTFR). The objective of this study was to assess the effects of S. cerevisiae (SC) on fiber degradation and rumen microbial populations in steers fed diets with different levels of dietary concentrate. Ten Simmental × Local crossbred steers (450 ± 50 kg BW) were assigned to a control group or an SC group. Both groups were fed the same basal diet but the SC group received SC supplementation (8 × 10(9) cfu/h/d through the ruminal fistula) following a two-period crossover design. Each period consisted of four phases, each of which lasted 17 d: 10 d for dietary adaptation, 6 d for degradation study, and 1 d for rumen sample collection. From the 1(st) to the 4(th) phase, steers were fed in a stepwise fashion with increasing CTFRs, i.e., 30:70, 50:50, 70:30, and 90:10. The kinetics of dry matter and fiber degradation of alfalfa pellets were evaluated; the rumen microbial populations were detected using real-time PCR. The results revealed no significant (P > 0.05) interactions between dietary CTFR and SC for most parameters. Dietary CTFR had a significant effect (P trend for these parameters. SC supplementation significantly (P trend of rumen fungi and protozoa in SC group (P < 0.1); copies of total bacteria in SC group were significantly higher (P < 0.05). Additionally, percentage of Ruminobacter amylophilus was significantly lower (P < 0.05) but percentage of Selenomonas ruminantium was significantly higher (P < 0.05) in the SC group. In a word, dietary CTFR had a significant effect on degradation characteristics of forage and rumen microbial population. S. cerevisiae had positive effects on DM and NDF degradation rate or effective degradability of forage; S. cerevisiae increased rumen total bacteria, fungi, protozoa, and lactate-utilizing bacteria but reduced

  5. Colorimetric determination of in vitro feed protein degradation

    African Journals Online (AJOL)

    bag technique. Diazotized f ish meal, maize gluten meal and soya-bean were incubated with strained rumen fluid and a suitable growth medium. Only 49o/o and 18% of the colour bound to soya-bean was bound to fish meal and corn gluten meal, respectively. Degradation values expressed as a percen- tage of the total ...

  6. Nutritive value and in situ rumen degradability of Marandu palisade grass at different locations within the pasture in a silvopastoral system with different babassu palm densities

    Directory of Open Access Journals (Sweden)

    Xerxes M. Tosta

    2015-09-01

    Full Text Available The objective of this study was to evaluate the nutritive value and in situ rumen degradability of grass collected from different locations within the pasture in a silvopastoral system with different densities of trees. The silvopastoral system consisted of Urochloa (syn. Brachiaria brizantha cv. Marandu and the babassu palm, Orbignya sp. (now: Attaleia speciosa. We used a completely randomized design with a 3 x 3 factorial arrangement for nutritional value (3 differently shaded locations and 3 palm tree densities and a 3 x 3 x 3 factorial arrangement for dry matter (DM disappearance (3 locations, 3 palm densities and 3 incubation times. There was no effect of location within the pasture nor of palm tree density on the concentrations of NDF, ADF, lignin, cellulose and hemicellulose. However, location influenced the concentrations of crude protein (CP and DM, with highest CP in material grown in full sunlight. At all densities, DM disappearance at 96 h for pasture grown in full sunlight exceeded that for pasture grown in full shade. These factors need to be compounded with the possible depressant effect of trees on DM production of pasture when considering the benefits of silvopastoral systems.Keywords: Digestibility, fiber, Northeast Brazil, protein, tree-grass associations, Urochloa  brizantha.DOI: 10.17138/TGFT(3187-193

  7. Rumen escape protein in grass and grass silage deterimened with a nylon bag and an enzymatic technique

    NARCIS (Netherlands)

    Cone, J.W.; Gelder, van A.H.; Mathijssen-Kamman, A.A.; Hindle, V.A.

    2004-01-01

    Rumen escape protein (REP) was determined for six grasses and 16 grass silages using a nylon bag technique and an in vitro technique using a proteolytic enzyme preparation of Streptomyces griseus. In vitro, the samples were incubated for 0, 1, 6 and 24 h. The highest correlation observed between

  8. Comparison of protein fermentation characteristics in rumen fluid determined with the gas production technique and the nylon bag technique

    NARCIS (Netherlands)

    Cone, J.W.; Rodrigues, M.A.M.; Guedes, C.M.; Blok, M.C.

    2009-01-01

    In this study, a modified version of the gas production technique was used to determine protein fermentation characteristics in rumen fluid of 19 feedstuffs. Performing the incubations in a N-free environment, and with an excess of rapidly fermentable carbohydrates, made N the limiting factor to

  9. Effects of rumen-undegradable protein on intake, performance, and mammary gland development in prepubertal and pubertal dairy heifers

    NARCIS (Netherlands)

    Silva, A.L.; Detmann, E.; Dijkstra, J.; Pedroso, A.M.; Silva, L.H.P.; Machado, A.F.; Sousa, F.C.; Santos, dos G.B.; Marcondes, M.I.

    2018-01-01

    The objective of this study was to evaluate the influence of different amounts of rumen-undegradable protein (RUP) on intake, N balance, performance, mammary gland development, carcass traits, and hormonal status of Holstein heifers at different physiological stages (PS). Sixteen prepubertal (PRE)

  10. Rumen microbial protein supply as estimated from purine derivative excretion on sheep receiving faba beans (vicia faba as supplement delivered at different feeding frequencies

    Directory of Open Access Journals (Sweden)

    Asmuddin Natsir

    2008-06-01

    Full Text Available Rapid and extensive degradation of faba beans (Vicia faba by ruminal microbes can result in substantial and undesirable N loss from the rumen. The purpose of this study was to test the hypothesis that offering faba beans as a supplement more than once a day to sheep receiving a combination of oaten chaff and lucerne chaff as a basal diet will increase microbial protein supply to the intestines. The experiment was conducted in a Latin square design (4 x 4 using four mature merino sheep. The treatments were: T0 = basal diet ad libitum + nil supplements, T1 = T0 + faba beans (FB fed once daily, T2 = T0 + FB fed twice daily, T3 = T0 + FB fed 8 times daily. The basal diet was given once per day at 09:00 in the morning while FB were given at the rate of approximately 0.5% of live body weight and delivered according to the treatment protocol. Urinary excretion of purine derivative (PD was used to estimate microbial protein supply. The results indicated that even though treatment statistically had no effects on total urine output, PD excretion in the urine, PD absorbed, estimated microbial N supply, and the efficiency of rumen microbial protein synthesis, provision of supplement to sheep numerically improved microbial N supply by 92% compared to that of control group. However, there were no differences within the supplemented group. Therefore, it is concluded that feeding faba beans more than once a day was unnecessary.

  11. In vitro rumen feed degradability assessed with DaisyII and batch culture: effect of sample size

    Directory of Open Access Journals (Sweden)

    Stefano Schiavon

    2010-01-01

    Full Text Available In vitro degradability with DaisyII (D equipment is commonly performed with 0.5g of feed sample into each filter bag. Literature reported that a reduction of the ratio of sample size to bag surface could facilitate the release of soluble or fine particulate. A reduction of sample size to 0.25 g could improve the correlation between the measurements provided by D and the conventional batch culture (BC. This hypothesis was screened by analysing the results of 2 trials. In trial 1, 7 feeds were incubated for 48h with rumen fluid (3 runs x 4 replications both with D (0.5g/bag and BC; the regressions between the mean values provided for the various feeds in each run by the 2 methods either for NDF (NDFd and in vitro true DM (IVTDMD degradability, had R2 of 0.75 and 0.92 and RSD of 10.9 and 4.8%, respectively. In trial 2, 4 feeds were incubated (2 runs x 8 replications with D (0.25 g/bag and BC; the corresponding regressions for NDFd and IVTDMD showed R2 of 0.94 and 0.98 and RSD of 3.0 and 1.3%, respectively. A sample size of 0.25 g improved the precision of the measurements obtained with D.

  12. Non-protein nitrogen and reduced phosphorus supply to sheep. 1. Phosphorus, volatile fatty acids, NH3 and pH of rumen liquid and daily fluid and phosphorus outflow from the rumen

    International Nuclear Information System (INIS)

    Hoeller, H.; Breves, G.; Martens, H.

    1983-01-01

    Sheep were fitted with cannulas in the rumen and in the abomasum and fed a straw/pellet diet providing 29% of total N as urea-N. The diet was deficient in P, and daily P intake during P depletion was about 1.3g. Phosphorus repletion was obtained in the same sheep by daily infusion of 2.85gP as phosphoric acid into the abomasum. During experimental periods, Cr-EDTA and 15 N-urea were continuously infused into the rumen for five days. During the last two days, blood samples and a total of ten samples from both rumen and abomasum were taken. Three depletion and three repletion experiments were performed. The following significant changes were noted in the P-deficient state (means): blood plasma inorganic P fell from 2.38 to 0.67mmol/L and rumen liquid total P from 5.65 to 1.71mmol/L, i.e. both levels decreased by approximately 70%. Fluid outflow from the rumen was raised by about 10% but P outflow decreased by about 68%. NH 3 concentrations in rumen fluid increased from 3.60 to 7.51mmol/L whereas total VFA concentration was reduced from 125 to 100mmol/L. Average pH values changed from 6.06 to 6.56. It is suggested that P depletion led to reduced microbial growth and activity in the rumen although data on microbial protein outflow from the rumen are not yet available. (author)

  13. Effect of diet and absence of protozoa on the rumen microbial community and on the representativeness of bacterial fractions used in the determination of microbial protein synthesis.

    Science.gov (United States)

    Belanche, A; de la Fuente, G; Pinloche, E; Newbold, C J; Balcells, J

    2012-11-01

    Accurate estimates of microbial synthesis in the rumen are vital to optimize ruminant nutrition. Liquid- (LAB) and solid-associated bacterial fractions (SAB) harvested from the rumen are generally considered as microbial references when microbial yield is calculated; however, factors that determine their composition are not completely understood. The aim of this study was to evaluate the effect of diet and absence or presence of rumen protozoa on the rumen microbial community. It was hypothesized that these treatments could modify the composition and representativeness of LAB and SAB. Twenty twin lambs (Ovis aries) were used; one-half of the twins were kept protozoa-free, and each respective twin sibling was faunated. At 6 mo of age, 5 animals from each group were randomly allocated to the experimental diets consisting of either alfalfa hay as the sole diet, or 50:50 mixed with ground barley grain. After 15 d of adaptation to the diet, animals were euthanized, rumen and abomasum contents were sampled, and LAB and SAB isolated. The presence of protozoa buffered the effect of diet on the rumen bacterial population. Faunated animals fed alfalfa hay had a greater abundance of F. succinogenes, anaerobic fungi and methanogens, as well as an enhanced rumen bacterial diversity. Cellulolytic bacteria were more abundant in SAB, whereas the abomasal abundance of most of the microorganisms studied was closer to those values observed in LAB. Rumen and abomasal samples showed similar bacterial DNA concentrations, but the fungal and protozoal DNA concentration in the abomasum was only 69% and 13% of that observed in the rumen, respectively, suggesting fungal and protozoal sequestration in the rumen or possible preferential degradation of fungal and protozoal DNA in the abomasum, or both. In conclusion, absence of protozoa and type of diet extensively modified the chemical composition of LAB and SAB as a consequence of changes in the microbial composition of these fractions.

  14. Study of the effect of presence or absence of protozoa on rumen fermentation and microbial protein contribution to the chyme.

    Science.gov (United States)

    Belanche, A; Abecia, L; Holtrop, G; Guada, J A; Castrillo, C; de la Fuente, G; Balcells, J

    2011-12-01

    The aim of this study was to investigate the effect of presence or absence of protozoa on rumen fermentation and efficiency of microbial protein synthesis under different diets. Of 20 twin paired lambs, 1 lamb of each pair was isolated from the ewe within 24 h after birth and reared in a protozoa-free environment (n = 10), whereas their respective twin-siblings remained with the ewe (faunated, n = 10). When lambs reached 6 mo of age, 5 animals of each group were randomly allocated to 1 of 2 experimental diets consisting of either alfalfa hay as the sole diet, or 50:50 mixed with ground barley grain according to a 2 × 2 factorial arrangement of treatments. After 15 d of adaptation to the diet, the animals were euthanized and total rumen and abomasal contents were sampled to estimate rumen microbial synthesis using C(31) alkane as flow marker. Different ((15)N and purine bases) and a novel (recombinant DNA sequences) microbial markers, combined with several microbial reference extracts (rumen protozoa, liquid and solid associated bacteria) were evaluated. Absence of rumen protozoa modified the rumen fermentation pattern and decreased total tract OM and NDF digestibility in 2.0 and 5.1 percentage points, respectively. The effect of defaunation on microbial N flow was weak, however, and was dependent on the microbial marker and microbial reference extract considered. Faunated lambs fed with mixed diet showed the greatest rumen protozoal concentration and the least efficient microbial protein synthesis (29% less than the other treatments), whereas protozoa-free lambs fed with mixed diet presented the smallest ammonia concentration and 34% greater efficiency of N utilization than the other treatments. Although (15)N gave the most precise estimates of microbial synthesis, the use of recombinant DNA sequences represents an alternative that allows separate quantification of the bacteria and protozoa contributions. This marker showed that presence of protozoa decrease the

  15. Effects of decreasing metabolizable protein and rumen-undegradable protein on milk production and composition and blood metabolites of Holstein dairy cows in early lactation.

    Science.gov (United States)

    Bahrami-Yekdangi, H; Khorvash, M; Ghorbani, G R; Alikhani, M; Jahanian, R; Kamalian, E

    2014-01-01

    This study was conducted to evaluate the effects of decreasing dietary protein and rumen-undegradable protein (RUP) on production performance, nitrogen retention, and nutrient digestibility in high-producing Holstein cows in early lactation. Twelve multiparous Holstein lactating cows (2 lactations; 50 ± 7 d in milk; 47 kg/d of milk production) were used in a Latin square design with 4 treatments and 3 replicates (cows). Treatments 1 to 4 consisted of diets containing 18, 17.2, 16.4, and 15.6% crude protein (CP), respectively, with the 18% CP diet considered the control group. Rumen-degradable protein levels were constant across the treatments (approximately 10.9% on a dry matter basis), whereas RUP was gradually decreased. All diets were calculated to supply a postruminal Lys:Met ratio of about 3:1. Dietary CP had no significant effects on milk production or milk composition. In fact, 16.4% dietary CP compared with 18% dietary CP led to higher milk production; however, this effect was not significant. Feed intake was higher for 16.4% CP than for 18% CP (25.7 vs. 24.3 kg/d). Control cows had greater CP and RUP intakes, which resulted in higher concentrations of plasma urea nitrogen and milk urea nitrogen; cows receiving 16.4 and 15.6% CP, respectively, exhibited lower concentrations of milk urea nitrogen (15.2 and 15.1 vs. 17.3 mg/dL). The control diet had a significant effect on predicted urinary N. Higher CP digestibility was recorded for 18% CP compared with the other diets. Decreasing CP and RUP to 15.6 and 4.6% of dietary dry matter, respectively, had no negative effects on milk production or composition when the amounts of Lys and Met and the Lys:Met ratio were balanced. Furthermore, decreasing CP and RUP to 16.4 and 5.4%, respectively, increased dry matter intake. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  16. 纳米硒对绵羊精饲料营养物质瘤胃降解率的影响%Impact of Nano-selenium on Sheep Rumen Degradability Rate of Concentrates Nutrition

    Institute of Scientific and Technical Information of China (English)

    石薇; 赵兴财; 任有蛇

    2011-01-01

    Using four sheep with body weight of (40 ± 2.0) kg, fitted with permanent rumen fistula, fed with mixed concentrate-based diets, and using of 4 X 4 Latin square design to study the nano-selenium diet (0, 0.3, 3, 6 g/d) of feed nutrients in sheep rumen degradation rate. The results show that: in 3 g/d group, concentrated feed crude protein (CP) ruminal degradation rate was significantly increased (P 0.05) with that of the control group; diet nano-selenium showed no significant difference (P >0.05) on dry matter of feed (DM), organic matter (OM) and rumen degradability. It suggested that the added dietary nano-selenium at level of 3 g/d played a significant role to promote rumen fermentatio, so it was appropriate to add at level of 3 g/d.%选用4只体质量为(40±2.0)kg,装有永久瘤胃瘘管的晋中绵羊,以羊草和混合精料为基础日粮,采用4×4拉丁方试验设计,研究日粮添加纳米硒(0,0.3,3,6g/d)对绵羊精饲料营养物质瘤胃降解率的影响.结果表明,日粮添加纳米硒3g/d的精饲料粗蛋白(CP)瘤胃降解率显著提高(P<0.05),日粮添加纳米硒6,0.3 g/d的精饲料粗蛋白(CP)瘤胃降解率与对照差异不显著(P>0.05);日粮添加纳米硒对精饲料干物质(DM)、有机物(OM)瘤胃降解率影响差异均不显著(P>0.05).这表明日粮纳米硒的添加水平为3 g/d时对瘤胃发酵有显著促进作用,故纳米硒的适宜添加水平为3g/d.

  17. ORGANIC MATTER AND CRUDE PROTEIN DEGRADATION SYNCHRONY IN DIETS SELECTED BY RANGE GOATS.

    Directory of Open Access Journals (Sweden)

    Rafael Ramírez Orduña

    2010-09-01

    Full Text Available The study was carried out with the aim to asses the synchrony of organic matter and crude protein degradation in the rumen of diets selected by range goats through two years. Five esophageal cannulated adult male goats were used to collect extrusa samples during summer (August 9–13 and autumn (November 29 –December 3 of 2006, winter (February 20 – 24, spring (April 29 –May 5, summer (September 10–15 and autumn (December 4–8 of 2007 and winter (February 20 – 25 and spring (May 9 –13 of 2008. Extrusa samples were subjected to chemical analysis to determine organic matter (OM, crude protein (CP in situ and in vitro true digestibility of dry matter. OM and CP intake were estimated by total fecal collection. Effective extent of degradation of the OM and CP was calculated hourly and total 24 hours. From the hourly quantity of OM and CP degraded, a synchrony index of CP to OM was calculated, and from the total 24 hours degradation, degraded organic matter intake and crude protein intake were also estimated. Sampling date was the main effect that determined the variation of diet OM and CP degradation parameters. Degraded crude protein intake as a proportion of degraded OM was affected by sampling date and was correlated to rainfall. During winter of the first year degraded crude protein intake was below the requirements for maintenance or to promote growth for range goats weighing 40 kg. Even though, synchrony index between OM and CP degradation was affected by sampling date goats maintained a high synchrony index throughout the years.

  18. A Systems Biology Approach Reveals Differences in the Dynamics of Colonization and Degradation of Grass vs. Hay by Rumen Microbes with Minor Effects of Vitamin E Supplementation

    Directory of Open Access Journals (Sweden)

    Alejandro Belanche

    2017-08-01

    Full Text Available Increasing the efficiency of utilization of fresh and preserved forage is a key target for ruminant science. Vitamin E is often used as additive to improve product quality but its impact of the rumen function is unknown. This study investigated the successional microbial colonization of ryegrass (GRA vs. ryegrass hay (HAY in presence of zero or 50 IU/d supplementary vitamin E, using a rumen simulation technique. A holistic approach was used to link the dynamics of feed degradation with the structure of the liquid-associated (LAB and solid-associated bacteria (SAB. Results showed that forage colonization by SAB was a tri-phasic process highly affected by the forage conservation method: Early colonization (0–2 h after feeding by rumen microbes was 2× faster for GRA than HAY diets and dominated by Lactobacillus and Prevotella which promoted increased levels of lactate (+56% and ammonia (+18%. HAY diets had lower DM degradation (-72% during this interval being Streptococcus particularly abundant. During secondary colonization (4–8 h the SAB community increased in size and decreased in diversity as the secondary colonizers took over (Pseudobutyrivibrio promoting the biggest differences in the metabolomics profile between diets. Secondary colonization was 3× slower for HAY vs. GRA diets, but this delay was compensated by a greater bacterial diversity (+197 OTUs and network complexity resulting in similar feed degradations. Tertiary colonization (>8 h consisted of a slowdown in the colonization process and simplification of the bacterial network. This slowdown was less evident for HAY diets which had higher levels of tertiary colonizers (Butyrivibrio and Ruminococcus and may explain the higher DM degradation (+52% during this interval. The LAB community was particularly active during the early fermentation of GRA and during the late fermentation for HAY diets indicating that the availability of nutrients in the liquid phase reflects the dynamics

  19. Measurement of the rate of bacterial protein synthesis in rumen. Part of a coordinated programme on tracer techniques in studies on the use of non-protein nitrogen in ruminants

    International Nuclear Information System (INIS)

    Leng, R.A.

    1975-10-01

    A technique has been developed to differentiate between rumen microbial and dietary protein at the duodenum. Mature whethers fitted with rumen, duodenal and ileal cannulas were fed protein poor diets with added urea, casein and formaldehyde treated casein. These contained 0.30, 1.54, 3.26 and 3.26 % N respectively. The isotopes 35 S or 15 N were infused into the rumen to label microbial protein so that microbial protein could be differentiated from dietary protein. By using the 35 S or 15 N system to mark microbes, it was found that 40% of the formaldehyde treated protein is fermented in the rumen and about 10% is lost in the feces. This indicates that approximately 50% of the protected protein was utilized by the sheep

  20. Evaluation of the furosine and homoarginine methods for determining reactive lysine in rumen-undegraded protein.

    Science.gov (United States)

    Boucher, S E; Pedersen, C; Stein, H H; Schwab, C G

    2009-08-01

    Three samples of soybean meal (SBM), 3 samples of expeller SBM (SoyPlus, West Central Cooperative, Ralston, IA), 5 samples of distillers dried grains with solubles (DDGS), and 5 samples of fish meal were used to evaluate the furosine and homoarginine procedures to estimate reactive Lys in the rumen-undegraded protein fraction (RUP-Lys). One sample each of SBM, expeller SBM, and DDGS were subjected to additional heat treatment in the lab to ensure there was a wide range in reactive RUP-Lys content among the samples. Furosine is a secondary product of the initial stages of the Maillard reaction and can be used to calculate blocked Lys. Homoarginine is formed via the reaction of reactive Lys with O-methylisourea and can be used to calculate the concentration of reactive Lys. In previous experiments, each sample was ruminally incubated in situ for 16 h, and standardized RUP-Lys digestibility of the samples was determined in cecectomized roosters. All rumen-undegraded residue (RUR) samples were analyzed for furosine and Lys; however, only 9 of the 16 samples contained furosine, and only the 4 unheated DDGS samples contained appreciable amounts of furosine. Blocked RUP-Lys was calculated from the furosine and Lys concentrations of the RUR. Both the intact feed and RUR samples were evaluated using the homoarginine method. All samples were incubated with an O-methylisourea/BaOH solution for 72 h and analyzed for Lys and homoarginine concentrations. Reactive Lys concentrations of the intact feeds and RUR were calculated. Results of the experiment indicate that blocked RUP-Lys determined via the furosine method was negatively correlated with standardized RUP-Lys digestibility, and reactive RUP-Lys determined via the guanidination method was positively correlated with standardized RUP-Lys digestibility. Reactive Lys concentrations of the intact samples were also highly correlated with RUP-Lys digestibility. In conclusion, the furosine assay is useful in predicting RUP

  1. Estimating rumen microbial protein supply for indigenous ruminants using nuclear and purine excretion techniques in Indonesia

    International Nuclear Information System (INIS)

    Soejono, M.; Yusiati, L.M.; Budhi, S.P.S.; Widyobroto, B.P.; Bachrudin, Z.

    1999-01-01

    The microbial protein supply to ruminants can be estimated based on the amount of purine derivatives (PD) excreted in the urine. Four experiments were conducted to evaluate the PD excretion method for Bali and Ongole cattle. In the first experiment, six male, two year old Bali cattle (Bos sondaicus) and six Ongole cattle (Bos indicus) of similar sex and age, were used to quantify the endogenous contribution to total PD excretion in the urine. In the second experiment, four cattle from each breed were used to examine the response of PD excretion to feed intake. 14 C-uric acid was injected in one single dose to define the partitioning ratio of renal:non-renal losses of plasma PD. The third experiment was conducted to examine the ratio of purine N:total N in mixed rumen microbial population. The fourth experiment measured the enzyme activities of blood, liver and intestinal tissues concerned with PD metabolism. The results of the first experiment showed that endogenous PD excretion was 145 ± 42.0 and 132 ± 20.0 μmol/kg W 0.75 /d, for Bali and Ongole cattle, respectively. The second experiment indicated that the proportion of plasma PD excreted in the urine of Bali and Ongole cattle was 0.78 and 0.77 respectively. Hence, the prediction of purine absorbed based on PD excretion can be stated as Y = 0.78 X + 0.145 W 0.75 and Y = 0.77 X + 0.132 W 0.75 for Bali and Ongole cattle, respectively. The third experiment showed that there were no differences in the ratio of purine N:total N in mixed rumen microbes of Bali and Ongole cattle (17% vs 18%). The last experiment, showed that intestinal xanthine oxidase activity of Bali cattle was lower than that of Ongole cattle (0.001 vs 0.015 μmol uric acid produced/min/g tissue) but xanthine oxidase activity in the blood and liver of Bali cattle was higher than that of Ongole cattle (3.48 vs 1.34 μmol/min/L plasma and 0.191 vs 0.131 μmol/min/g liver tissue). Thus, there was no difference in PD excretion between these two breeds

  2. AAA-ATPases in Protein Degradation

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    Ravikiran S. Yedidi

    2017-06-01

    Full Text Available Proteolytic machineries containing multisubunit protease complexes and AAA-ATPases play a key role in protein quality control and the regulation of protein homeostasis. In these protein degradation machineries, the proteolytically active sites are formed by either threonines or serines which are buried inside interior cavities of cylinder-shaped complexes. In eukaryotic cells, the proteasome is the most prominent protease complex harboring AAA-ATPases. To degrade protein substrates, the gates of the axial entry ports of the protease need to be open. Gate opening is accomplished by AAA-ATPases, which form a hexameric ring flanking the entry ports of the protease. Protein substrates with unstructured domains can loop into the entry ports without the assistance of AAA-ATPases. However, folded proteins require the action of AAA-ATPases to unveil an unstructured terminus or domain. Cycles of ATP binding/hydrolysis fuel the unfolding of protein substrates which are gripped by loops lining up the central pore of the AAA-ATPase ring. The AAA-ATPases pull on the unfolded polypeptide chain for translocation into the proteolytic cavity of the protease. Conformational changes within the AAA-ATPase ring and the adjacent protease chamber create a peristaltic movement for substrate degradation. The review focuses on new technologies toward the understanding of the function and structure of AAA-ATPases to achieve substrate recognition, unfolding and translocation into proteasomes in yeast and mammalian cells and into proteasome-equivalent proteases in bacteria and archaea.

  3. AAA-ATPases in Protein Degradation.

    Science.gov (United States)

    Yedidi, Ravikiran S; Wendler, Petra; Enenkel, Cordula

    2017-01-01

    Proteolytic machineries containing multisubunit protease complexes and AAA-ATPases play a key role in protein quality control and the regulation of protein homeostasis. In these protein degradation machineries, the proteolytically active sites are formed by either threonines or serines which are buried inside interior cavities of cylinder-shaped complexes. In eukaryotic cells, the proteasome is the most prominent protease complex harboring AAA-ATPases. To degrade protein substrates, the gates of the axial entry ports of the protease need to be open. Gate opening is accomplished by AAA-ATPases, which form a hexameric ring flanking the entry ports of the protease. Protein substrates with unstructured domains can loop into the entry ports without the assistance of AAA-ATPases. However, folded proteins require the action of AAA-ATPases to unveil an unstructured terminus or domain. Cycles of ATP binding/hydrolysis fuel the unfolding of protein substrates which are gripped by loops lining up the central pore of the AAA-ATPase ring. The AAA-ATPases pull on the unfolded polypeptide chain for translocation into the proteolytic cavity of the protease. Conformational changes within the AAA-ATPase ring and the adjacent protease chamber create a peristaltic movement for substrate degradation. The review focuses on new technologies toward the understanding of the function and structure of AAA-ATPases to achieve substrate recognition, unfolding and translocation into proteasomes in yeast and mammalian cells and into proteasome-equivalent proteases in bacteria and archaea.

  4. Estimation of rumen microbial protein production from urinary purine derivatives in zebu cattle and water buffalo

    International Nuclear Information System (INIS)

    Liang, J.B.; Pimpa, O.; Abdullah, N.; Jelan, Z.A.; Nolan, J.V.

    1999-01-01

    Two experiments were conducted in order to develop equations for predicting rumen microbial protein production for indigenous Kedah-Kelantan (KK) cattle and swamp buffaloes in Malaysia, using urinary purine derivatives (PD) excretion rates. Endogenous PD excretion rates determined by a fasting procedure for KK cattle and swamp buffalo were 275 and 370 μmol/kg W 0.75 /day, respectively. Urinary PD excretion rate per kg digestible organic matter intake (DOMI) for KK cattle was higher than that for swamp buffalo, reconfirming the earlier findings. Glomerular filtration rate, allantoin and uric acid tubular load and PD re-absorption rate for swamp buffalo were generally higher than those for KK cattle. However, due to the large variations among animals within species, these parameters were not significantly different between species. Nevertheless, the higher PD reabsorption in swamp buffalo provides support for the earlier postulation that the lower urinary PD excretion rate of swamp buffalo was due to their higher recycling of plasma PD as compared to KK cattle. Labelled 8- 14 C uric acid was used to estimate the ratio of renal to non-renal PD excretion. The recovery rates of the radioactive tracer via the renal route for both species were much lower than values reported previously for unlabelled PD for European cattle. (author)

  5. Uptake and degradation of circulating proteins by the liver.

    NARCIS (Netherlands)

    Buys, Carolus Henricus Cornelis Maria

    1976-01-01

    Circulating proteins, like all proteins in a living animal, are subject to continual replacement or turnover. This process implies both synthesis and degradation, This thesis deals with the degradative part of turnover of circulating proteins. ... Summary

  6. Precision and accuracy of the NDF rumen degradability of hays measured by the Daisy fermenter

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    C. Zanfi

    2010-04-01

    Full Text Available An inventory of 162 hay samples from Austrian permanent grasslands was used to obtain information about the precision of the in vitro NDF degradability (NDFd measured by the Daisy fermenter and its accuracy to predict in situ NDFd. The within forage standard error of the in vitro NDFd triplicate, obtained in five consecutive incubations, was equal to 2.8%, while the effect of the four jar positions in the fermenter was not significant. The cutting frequency had a great impact on the in situ effective NDFd of hays, which ranged (P<0.01 from values of 32.9, 43.1 and 48.3% in hays obtained from 2, 3 and 4 cuts/season, respectively. The regression analysis between the in vitro and in situ NDFd values (measured at 48h and effective, k=3%/h allowed to obtain medium degrees of correlation (r2 = 0.69 – 0.71; P<0.01 and low levels of accuracy (RSE = 4.0 -4.6 %.

  7. Investigations on the effect of forage source, grinding, and urea supplementation on ruminal fermentation and microbial protein flow in a semi-continuous rumen simulation system.

    Science.gov (United States)

    Hildebrand, Bastian; Boguhn, Jeannette; Rodehutscord, Markus

    2011-10-01

    The objective of the present study was to compare the effect of maize silage and grass silage on microbial fermentation and protein flow in a semi-continuous rumen simulation system (Rusitec) when milling screen size (MSS) during grinding was varied. Oven-dried silages were milled through screens of 1, 4 or 9 mm pore size and incubated for 48 h in a Rusitec system. Furthermore, the effect of N supplementation to maize silage (MSS: 4 mm) was investigated and single dose vs. continuous infusion of urea-N were compared. Degradation of organic matter (OM), crude protein (CP), fibre fractions and non-structural carbohydrates (NSC) as well as short-chain fatty acid production differed significantly between forage sources. Urea-N supplementation improved the degradation of NSC, but not that of fibre fractions in maize silage. The way of urea supply had only marginal effects on fermentation characteristics. An increase in MSS, and consequently in mean feed particle size, led to an improvement in the degradation of OM, CP and NSC, but efficiency of microbial net protein synthesis (EMPS; mg microbial N flow/g degraded OM) and the microbial amino acid profile were less affected. EMPS was higher in grass silage than in maize silage and was improved by urea-N supplementation in maize silage. This study indicates that fermentation of NSC as well as EMPS during incubation of maize silage was limited by availability of NH3-N. Furthermore, an increase in MSS above 1 mm seems to improve fermentation of silages in the Rusitec system.

  8. Microbial degradation of dissolved proteins in seawater

    International Nuclear Information System (INIS)

    Hollibaugh, J.T.; Azam, F.

    1983-01-01

    An experimental protocol using radiolabeled proteins was developed to investigate the rates and mechanisms whereby dissolved proteins are degraded in natural marine plankton communities. The results of field observations and laboratory experiments indicate that proteins are degraded by a particle-bound, thermolabile system, presumably bacteria-associated enzymes, with an apparent half-saturation constant of ca. 25 μg bovine serum albumin (BSA) per liter. Gel permeation chromatography indicated that peptides of chain length intermediate between BSA and the final products of degradation (MW<700) do not accumulate in the medium. Competition experiments indicate that the system is relatively nonspecific. Turnover rates for the protein pool in samples collected in the Southern California Bight were of the same order of magnitude as the turnover rate of the L-leucine pool and were correlated with primary productivity, chlorophyll a concentrations, bacterial abundance and biomass, and L-leucine turnover rate. These data suggest that amino acids derived from proteins are utilized preferentially and do not completely mix with the amino acids in the bulk phase

  9. Caracterização, fracionamento protéico, degradabilidade ruminal e digestibilidade in vitro da matéria seca e proteína bruta do resíduo de cervejaria úmido e fermentado - DOI: 10.4025/actascianimsci.v29i3.558 Characterization, protein fractioning, dry matter and crude protein rumen degradability and in vitro digestibility of wet and fermented brewer’s grain - DOI: 10.4025/actascianimsci.v29i3.558

    Directory of Open Access Journals (Sweden)

    Jacco Arnoud Erke

    2007-12-01

    Full Text Available Foram avaliadas as frações da proteína e dos carboidratos, a degradabilidade ruminal efetiva (DE da matéria seca (MS e proteína bruta (PB, a digestibilidade ruminal in vitro (DRIV da MS e PB, a digestibilidade intestinal in vitro (DIIV da proteína nãodegradada no rúmen (PNDR e os perfis de aminoácidos (AA e de ácidos graxos (AG do resíduo de cervejaria úmido (RCU e fermentado (RCF. O RCF foi obtido pelo processo de fermentação microbiana do RCU. Para determinar a DE da MS e PB do RCU e RCF, foram utilizados três novilhos da raça Holandesa, portadores de cânula ruminal. A DIIV da PNDR foi obtida pelo método de três estágios. Os dados obtidos para DE da MS e PB foram submetidos à análise de variância, em delineamento inteiramente casualizado. A fração A da PB do RCU foi de 7,9% e do RCF de 13,1% da PB. A DE da PB a 5% h-1 não diferiu (p IV da PB do RCF foi de 8,7% e as DIIV da PNDR do RCU e do RCF foram de 70,5 e 72,5%, respectivamente. Os perfis de AA e AG do RCU e RCF foram similares. O processo de fermentação anaeróbico não alterou as características nutricionais do RCU.The study evaluated the protein and carbohydrate fraction, dry matter (DM and crude protein (CP effective rumen degradability (ED, DM and CP in vitro ruminal digestibility (RDIV, rumen-undegradable protein (RUDP in vitro intestinal digestion (IDIV and amino-acid (AA and fatty acid (FA profile of the wet brewer’s grain (WBG, and fermented brewer’s grain (FBG. FBG was obtained from WBG fermentation. The DM and CP ED of WBG and FBG were determined in three Holstein steers with ruminal cannula. The IDIV of RUDP was obtained by the three-stage method. The values obtained for DM and CP ED were submitted to variance analysis, in a randomized design. The A fraction of WBG CP was 7.9%, and for FBG 13.1% of CP. The CP RD in a rate of 5% h-1 did not differ (p > 0.05 between WBG and FBG. The crude protein RDIV of FBG was 8.7% and IDIV of RUDP of WBG and

  10. Performance and metabolite profile of dairy cows fed tropical grasses and concentrates containing crude protein with low or high degradability

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    Raphael dos Santos Gomes

    Full Text Available ABSTRACT Ten Holstein-Zebu crossbred cows distributed into two simultaneous Latin squares (5 × 5 as a 2 × 2 factorial arrangement formed by chopped sugarcane or elephant grass silage, both with high or low protein degradability supplements and a corn silage as a control treatment, were compared using orthogonal contrasts. The studied variables were the performance, plasma concentrations of urea-N, glucose, and creatinine, urine-N and milk urea-N, and the nychthemeral variation in NH3-N in the rumen fluid of dairy cows. Nutrient intake, milk production, and milk composition were affected by the treatments. The total mixed ration containing elephant grass silage combined with rumen undegradable protein (RUP provided balanced amounts of carbon and nitrogen in the rumen. This effect may explain the 18% increase in milk yield compared with the other treatments. The diurnal pattern of ruminal NH3-N was interpreted with a sinusoid model. In general, cows fed elephant grass silage exhibited higher concentrations of blood plasma and milk urea-N than animals fed sugarcane. The cows that consumed elephant grass silage with rumen degradable protein concentrate showed a higher milk urea-N compared with animals that consumed elephant grass silage with the RUP concentrate. The use of diets based on corn silage leads to a better use of nitrogen compounds because these diets resulted in lower levels of urea-N in the plasma, urine, and milk at the same level of milk production compared with diets containing elephant grass silage or chopped sugarcane as roughages. In sugarcane-based diets, even greater nitrogen losses in the urine are observed, despite the presence of readily fermentable carbohydrates in the diet.

  11. Rumen Microbial Protein Production in Rumen-Simulating-Technique (RUSITEC) Using 15N-Urea Nitrogen, as Influenced By Hay and Barley Ratios in Feed

    International Nuclear Information System (INIS)

    Al-Masri, M. R.; Abel, HJ.; Steinberg, W.

    2004-01-01

    Metabolism of dietary nitrogen using labeled 15 N and the changes in the microbial protein mass and NH3-N were studied in five rumen-simulating-technique(RUSITEC)-fermenters, which were run simultaneously in three identically repeated experiments. Each experiment consisted of a 6-day adaptation period followed directly by a 3-day collection period. The feed of the fermenters (G1, G2, G3, G4 and G5) varied in the ratio of barley. The barley increased by 20% between the fermenters. Grass hay+barley (g/d) in the feed of the fermenters was 10+2 (G1), 8+4 (G2), 6+6 (G3), 4+8 (G4) and 2+10 (G5). The results indicated that there were no significant (P>0.05) changes in the amounts of microbial nitrogen (92-118 mg/d) and microbial mass syntheses which were (mg/d): 1154 (G1), 1063 (G2), 1152 (G3), 1127 (G4) and 1362 (G5). Increasing the proportion of barley in the fermenters (G4 and G5) decreased NH3-N amounts (G2 and G3) significantly (P<0.05). The energy was not efficiently used in G5 having a lower ratio of the microbial nitrogen and microbial mass to the total short chain fatty acids than that other fermenters. (authors)

  12. Ninety-nine de novo assembled genomes from the moose (Alces alces) rumen microbiome provide new insights into microbial plant biomass degradation

    Science.gov (United States)

    Svartström, Olov; Alneberg, Johannes; Terrapon, Nicolas; Lombard, Vincent; de Bruijn, Ino; Malmsten, Jonas; Dalin, Ann-Marie; Muller, Emilie E.L.; Shah, Pranjul; Wilmes, Paul; Henrissat, Bernard; Aspeborg, Henrik; Andersson, Anders F.

    2017-01-01

    The moose (Alces alces) is a ruminant that harvests energy from fiber-rich lignocellulose material through carbohydrate-active enzymes (CAZymes) produced by its rumen microbes. We applied shotgun metagenomics to rumen contents from six moose to obtain insights into this microbiome. Following binning, 99 metagenome-assembled genomes (MAGs) belonging to eleven prokaryotic phyla were reconstructed and characterized based on phylogeny and CAZyme profile. The taxonomy of these MAGs reflected the overall composition of the metagenome, with dominance of the phyla Bacteroidetes and Firmicutes. Unlike in other ruminants, Spirochaetes constituted a significant proportion of the community and our analyses indicate that the corresponding strains are primarily pectin digesters. Pectin-degrading genes were also common in MAGs of Ruminococcus, Fibrobacteres and Bacteroidetes, and were overall overrepresented in the moose microbiome compared to other ruminants. Phylogenomic analyses revealed several clades within the Bacteriodetes without previously characterized genomes. Several of these MAGs encoded a large numbers of dockerins, a module usually associated with cellulosomes. The Bacteroidetes dockerins were often linked to CAZymes and sometimes encoded inside polysaccharide utilization loci (PULs), which has never been reported before. The almost one hundred CAZyme-annotated genomes reconstructed in this study provides an in-depth view of an efficient lignocellulose-degrading microbiome and prospects for developing enzyme technology for biorefineries. PMID:28731473

  13. Protein degradation and protection against misfolded or damaged proteins

    Science.gov (United States)

    Goldberg, Alfred L.

    2003-12-01

    The ultimate mechanism that cells use to ensure the quality of intracellular proteins is the selective destruction of misfolded or damaged polypeptides. In eukaryotic cells, the large ATP-dependent proteolytic machine, the 26S proteasome, prevents the accumulation of non-functional, potentially toxic proteins. This process is of particular importance in protecting cells against harsh conditions (for example, heat shock or oxidative stress) and in a variety of diseases (for example, cystic fibrosis and the major neurodegenerative diseases). A full understanding of the pathogenesis of the protein-folding diseases will require greater knowledge of how misfolded proteins are recognized and selectively degraded.

  14. Evaluation of nutritive value of palm kernel cake fermented with molds as source of protein in ruminant

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    Wisri Puastuti

    2014-06-01

    Full Text Available The objective of the research was to improve the nutritive value of PKC through fermentation and to evaluate its degradation characteristics in the rumen and post rumen digestibility as a protein feed source for ruminants. PKC was fermented using Aspergillus niger, Trichoderma viridae and Aspergillus oryzae. To evaluate the in sacco rumen degradability, 2 rumen fistulated females Fries Holstein 3.5 years old were used. Samples were incubated in the rumen for 0, 4, 8, 12, 16, 24, 48 and 72 hours. Determination of dry matter (DM degradation characteristics value and crude protein (CP in the rumen was calculated based on formula y = a + b (1 - e-ct. The experiments were conducted using a completely randomized design with four replicates. The results showed that fermentation increased protein content of the PKC by 79.21% with the highest increase from fermentation using Aspergillus oryzae (88.34%. DM and CP degradability ​in the rumen and post rumen of fermented PKC was affected by type of mold used for fermentation. Fermentation increased the amount of water soluble DM (a of fermented PKC with average of 46.7%, but the value of insoluble but degradable fraction in the rumen (b was decreased. Fermentation by molds resulited in the reduction of fraction of insoluble CP but degradable (b in the rumen by 50.42%. PKC fermentation by Aspergillus oryzae resulted in the higest CP degradability in the rumen and post rumen. It can be concluded that PKC has a high content of degradable CP in the rumen even without fermentation. Protein source from PKC fermented using Aspergillus oryzae categorized as the best source of feed protein in terms of increasing CP content and digestibility.

  15. Influence of stage of maturity of grass silages on protein digestion and microbial protein synthesis in the rumen.

    NARCIS (Netherlands)

    Bosch, M.W.; Bruchem, van J.; Bongers, L.J.G.M.; Tamminga, S.

    1994-01-01

    In four change-over experiments, wilted grass silages, differing in growth stage at harvesting and as a consequence in cell wall content, were fed ad libitum to dairy cows in early and late lactation. Ruminal degradation rate of the crude protein fraction of the silages was investigated using nylon

  16. Comparison of three 15N methods to correct for microbial contamination when assessing in situ protein degradability of fresh forages.

    Science.gov (United States)

    Kamoun, M; Ammar, H; Théwis, A; Beckers, Y; France, J; López, S

    2014-11-01

    The use of stable (15)N as a marker to determine microbial contamination in nylon bag incubation residues to estimate protein degradability was investigated. Three methods using (15)N were compared: (15)N-labeled forage (dilution method, LF), (15)N enrichment of rumen solids-associated bacteria (SAB), and (15)N enrichment of rumen liquid-associated bacteria (LAB). Herbage from forages differing in protein and fiber contents (early-cut Italian ryegrass, late-cut Italian ryegrass, and red clover) were freeze-dried and ground and then incubated in situ in the rumen of 3 steers for 3, 6, 12, 24, and 48 h using the nylon bag technique. The (15)N-labeled forages were obtained by fertilizing the plots where herbage was grown with (15)NH4 (15)NO3. Unlabeled forages (obtained from plots fertilized with NH4NO3) were incubated at the same time that ((15)NH4)2SO4 was continuously infused into the rumen of the steers, and then pellets of labeled SAB and LAB were isolated by differential centrifugation of samples of ruminal contents. The proportion of bacterial N in the incubation residues increased from 0.09 and 0.45 g bacterial N/g total N at 3 h of incubation to 0.37 and 0.85 g bacterial N/g total N at 48 h of incubation for early-cut and late-cut ryegrass, respectively. There were differences (P forage (late-cut ryegrass) was 0.51, whereas the corrected values were 0.85, 0.84, and 0.77 for the LF, SAB, and LAB methods, respectively. With early-cut ryegrass and red clover, the differences between uncorrected and corrected values ranged between 6% and 13%, with small differences among the labeling methods. Generally, methods using labeled forage or labeled SAB and LAB provided similar corrected degradability values. The accuracy in estimating the extent of degradation of protein in the rumen from in situ disappearance curves is improved when values are corrected for microbial contamination of the bag residue.

  17. Degradabilidade do capim-elefante (Pennisetum purpureum, Schum. e da cana-de-açúcar (Saccharum officinarum, L. mais uréia no rúmen de vacas mestiças Holandês × Zebu em lactação Rumen degradability of elephantgrass (Pennisetum purpureum, Schum. and sugarcane (Saccharum officinarum, L. plus urea in crossbred lactating cows

    Directory of Open Access Journals (Sweden)

    F.C.F. Lopes

    1999-08-01

    Full Text Available Elephantgrass shows a high stocking rate only during the rainy season. Forage production is drastically reduced in the dry season. One of the alternatives to minimize this problem is the supplementation with sugarcane plus urea. In this trial the main goal was to estimate the dry matter intake (DMI of elephantgrass, chopped sugarcane plus 1% urea and their degradability in three lactating (11.6kg/milk/day rumen fistulated crossbred cows (472kg/L.W. grazing elephantgrass during the dry season. The cows had access during three consecutive night to the paddocks grazed every 30 days in a stocking rate, of 4.5 cows/hectare. Between the two milking times, cows received chopped sugarcane plus 1% urea. The DMI was on average 1.01%/L.W. for the elephantgrass and 0.88%/L.W. for the sugarcane plus 1% urea. Total DMI, taking into account both roughage and concentrate (2kg/cow/day was 2.25%/L.W. The effective rumen degradability (ED, considering the rate of passage in the rumen of 5%/hour was 44.46% for elephantgrass and 41.94% for sugarcane plus 1% urea, DM. The elephantgrass ED crude protein was 48.58% and its neutral detergent fiber 33.82%. During the dry season lactating crossbred cows grazing elephantgrass have to receive a roughage supplement and concentrate to produce around to 12kg of milk per day.

  18. The development of an intraruminal nylon bag technique using non-fistulated animals to assess the rumen degradability of dietary plant materials.

    Science.gov (United States)

    Pagella, J H; Mayes, R W; Pérez-Barbería, F J; Ørskov, E R

    2018-01-01

    Although the conventional in situ ruminal degradability method is a relevant tool to describe the nutritional value of ruminant feeds, its need for rumen-fistulated animals may impose a restriction on its use when considering animal welfare issues and cost. The aim of the present work was to develop a ruminal degradability technique which avoids using surgically prepared animals. The concept was to orally dose a series of porous bags containing the test feeds at different times before slaughter, when the bags would be removed from the rumen for degradation measurement. Bags, smaller than those used in the conventional nylon bag technique, were made from woven nylon fabric, following two shape designs (rectangular flat shape, tetrahedral shape) and were fitted with one of three types of device for preventing their regurgitation. These bags were used in two experiments with individually housed non-pregnant, non-lactating sheep, as host animals for the in situ ruminal incubation of forage substrates. The bags were closed at the top edge by machine stitching and wrapped in tissue paper before oral dosing. Standard times for ruminal incubation of substrates in all of the tests were 4, 8, 16, 24, 48, 72 and 96 h before slaughter. The purpose of the first experiment was to compare the effectiveness of the three anti-regurgitation device designs, constructed from nylon cable ties ('Z-shaped', ARD1; 'double Z-shaped', ARD2; 'umbrella-shaped', ARD3), and to observe whether viable degradation curves could be generated using grass hay as the substrate. In the second experiment, three other substrates (perennial ryegrass, red clover and barley straw) were compared using flat and tetrahedral bags fitted with type ARD1 anti-regurgitation devices. Non-linear mixed-effect regression models were used to fit asymptotic exponential curves of the percentage dry matter loss of the four substrates against time of incubation in the reticulorumen, and the effect of type of anti

  19. Degradability of dry matter and crude protein of dry grains and wet grain silages from different processing corn hybrids (Zea mays)

    OpenAIRE

    Wagner dos Reis; Ciniro Costa; Paulo Roberto de Lima Meirelles; Marina Gabriela Berchiol da Silva; Marco Aurélio Factori; Janaína Conte Hadlich; Kátia de Oliveira; Erikelly Aline Ribeiro de Santana; Cristiano Magalhães Pariz; Josineudson Augusto II de Vasconcelos Silva

    2013-01-01

    The objective of this work was to evaluate the effect of processing two corn hybrids conserved, dry and humid grains, the dry matter (DM) and crude protein (CP) degradability in situ. The particle size was determined and difference was verified in MGD (Medium Geometric Diameter) of processed ingredients. Three sheep were used with rumen canulated, in a completely randomized design, using a factorial outline 2 x 2 x 3, being two corn hybrid, two conservation methods and three processing forms ...

  20. Effects of rumen undegradable protein supplementation on productive performance and indicators of protein and energy metabolism in Holstein fresh cows.

    Science.gov (United States)

    Amanlou, H; Farahani, T Amirabadi; Farsuni, N Eslamian

    2017-05-01

    The objective of this study was to determine the effects of feeding increased dietary crude protein (CP) on productive performance and indicators of protein and energy metabolism during 21 d postpartum. Thirty multiparous Holstein dairy cows were balanced by previous lactation milk yield, body condition score (BCS) at calving, and parity and randomly allocated to 1 of 3 dietary treatments from calving until 21 d postpartum. Dietary treatments were 16.0% CP with 5.0% rumen undegradable protein (RUP) based on dry matter (DM) (16CP), 18.7% CP with 7.0% RUP based on DM (19CP), and 21.4% CP with 9.0% RUP based on DM (21CP). Diets were similar in net energy for lactation (approximately 1.7 Mcal/kg of DM) and CP levels were increased with corn gluten meal and fish meal. Dry matter intake (DMI) was increased by increasing dietary CP levels from 16.0 to 19.0% of DM, but dietary CP beyond 19.0% had no effect on DMI. Milk yields were 4.7 and 6.5 kg/d greater in cows fed the 19CP and 21CP diets versus those fed the 16CP diet, whereas 4% fat-corrected milk was greater for cows fed the 21CP than the 16CP diet (36.0 vs. 31.4 kg/d). Milk protein content and yield, lactose yield, and milk urea nitrogen were elevated by increased dietary CP. Milk lactose content and fat yield were not different among dietary treatments, but milk fat content tended to decline with increasing content of CP in diets. High CP levels increased milk N secretion but decreased milk N efficiency. Apparent digestibility of DM, CP, and neutral detergent fiber was greater on the 19CP and 21CP diets compared with the 16CP diet. Cows fed the 19CP and 21CP diets lost less body condition relative to those fed the 16CP diet over 21 d postpartum. Feeding higher CP levels increased the concentrations of serum albumin, albumin to globulin ratio, and urea nitrogen and decreased aspartate aminotransferase, nonesterified fatty acids, and β-hydroxybutyrate, but had no effect on globulin, glucose, cholesterol, or

  1. Non-degradative Ubiquitination of Protein Kinases.

    Directory of Open Access Journals (Sweden)

    K Aurelia Ball

    2016-06-01

    Full Text Available Growing evidence supports other regulatory roles for protein ubiquitination in addition to serving as a tag for proteasomal degradation. In contrast to other common post-translational modifications, such as phosphorylation, little is known about how non-degradative ubiquitination modulates protein structure, dynamics, and function. Due to the wealth of knowledge concerning protein kinase structure and regulation, we examined kinase ubiquitination using ubiquitin remnant immunoaffinity enrichment and quantitative mass spectrometry to identify ubiquitinated kinases and the sites of ubiquitination in Jurkat and HEK293 cells. We find that, unlike phosphorylation, ubiquitination most commonly occurs in structured domains, and on the kinase domain, ubiquitination is concentrated in regions known to be important for regulating activity. We hypothesized that ubiquitination, like other post-translational modifications, may alter the conformational equilibrium of the modified protein. We chose one human kinase, ZAP-70, to simulate using molecular dynamics with and without a monoubiquitin modification. In Jurkat cells, ZAP-70 is ubiquitinated at several sites that are not sensitive to proteasome inhibition and thus may have other regulatory roles. Our simulations show that ubiquitination influences the conformational ensemble of ZAP-70 in a site-dependent manner. When monoubiquitinated at K377, near the C-helix, the active conformation of the ZAP-70 C-helix is disrupted. In contrast, when monoubiquitinated at K476, near the kinase hinge region, an active-like ZAP-70 C-helix conformation is stabilized. These results lead to testable hypotheses that ubiquitination directly modulates kinase activity, and that ubiquitination is likely to alter structure, dynamics, and function in other protein classes as well.

  2. Effect of N:S ratio on the corporation of 35S into the thioamino acids of the microbial protein in an in vitro rumen system

    International Nuclear Information System (INIS)

    Walli, T.K.; Mudgal, V.D.

    1978-01-01

    Three fistulated Tharparkar cows and three fistulated Murrah buffaloes, fed with three isonitrogenous urea based concentrate mixtures with nitrogen : sulphur ratio of 20:1, 10:1 and 5:1, alongwith wheat straw, were used as donors of rumen inoculum for an in vitro experiment. The incubation vessels contained either of the three urea based purified substrates of N:S ratios (20:1, 10:1 and 5:1). Three μCi of 35 S labelled Na 2 SO 4 was added to each incubation vessel. At the end of 24 h incubation, the proteins were precipitated with TCA. The proteins were further hydrolysed for the estimation of thio-amino acids, namely cystine and methionine. 35 S radioactivity in the protein precipitate and also in the cystine and methionine fraction was measured by liquid scintillation. From the present studies it could be suggested that the dietary nitrogen: sulphur ratio of 10:1 is optimum for the maximum protein and thioamino acid synthesis in the rumen. The studies further revealed that the buffalo rumen microbes synthesise more cystine and methionine and consequently more proteins than the microbes in cow rumen. (author)

  3. Effect of niacin supplementation on rumen fermentation characteristics and nutrient flow at the duodenum in lactating dairy cows fed a diet with a negative rumen nitrogen balance.

    Science.gov (United States)

    Aschemann, Martina; Lebzien, Peter; Hüther, Liane; Südekum, Karl-Heinz; Dänicke, Sven

    2012-08-01

    The aim of the present experiment was to ascertain if a daily niacin supplementation of 6 g/cow to lactating dairy cow diets can compensate for the decrease in rumen microbial fermentation due to a negative rumen nitrogen balance (RNB). A total of nine ruminally and duodenally fistulated lactating multiparous German Holstein cows was used. The diets consisted of 10 kg dry matter (DM) maize silage and 7 kg DM concentrate and differed as follows: (i) Diet RNB- (n = 6) with energy and utilisable crude protein (CP) at the duodenum (uCP) according to the average requirement of the animals, but with a negative RNB (-0.41 g N/MJ metabolisable energy [ME]); (ii) Diet RNB0 (n = 7) with energy, uCP, and RNB (0.08 g N/MJ ME) according to the average requirement of the animals; and (iii) Diet NA (nicotinic acid; n = 5), which was the same diet as RNB-, but supplemented with 6 g niacin/d. The negative RNB affected the rumen fermentation pattern and reduced ammonia content in rumen fluid and the daily duodenal flows of microbial CP (MP) and uCP. Niacin supplementation increased the apparent ruminal digestibility of neutral detergent fibre. The efficiency of microbial protein synthesis per unit of rumen degradable CP was higher, whereby the amount of MP reaching the duodenum was unaffected by niacin supplementation. The number of protozoa in rumen fluid was higher in NA treatment. The results indicated a more efficient use of rumen degradable N due to changes in the microbial population in the rumen when niacin was supplemented to diets deficient in RNB for lactating dairy cows.

  4. Pengaruh Nisbah Energi-Protein, Nitrogen-Sulfur dan Kalsium-Fosfor Terhadap Produk Metabolisme Rumen dan Kecernaan Substrat

    Directory of Open Access Journals (Sweden)

    S.N.O. Suwandyastuti

    2013-10-01

    Full Text Available Influence of the ratio of energy-protein, nitrogen-sulphur, and calsium-phosphor upon rumen metabolism product and digestibility of substrat ABSTRACT. The Rumen microbes are capable to digest the glucosa polymer of plant waste for energy source and can used the Non Protein Nitrogen (NPN for body protein synthesis, if the other precussor (Sulphur, Phospor and Branch Chain of Carbon are available.  To know the effectivity of the utilization of plant waste for energy, an experiment have been conducted by in vitro method, used the Randomized Black Design, four replication, factorial 33.  The factors tested are : (1 three levels of energy : protein (E/P ratio : 4, 5 and 6; (2 three levels of Nitrogen : Sulphur (N/Sratio: 7.5, 10 and 12.5; (3 three levels of Calsium : Phospor (Ca/P ratio : 0.5, 1 and 2. The variables measured are : synthesis of protein microbes (SPM ; production of Volatile Fatty Acid (VFA and Nitrogen Ammonia (N-NH3; the digestibility of substrat. Based on the all variable measured, the experiment can be concluded : (1 the effectivity of the utilization of rice straw will be increased if it used is fortified with 50 percent TDN as energy source (E/P=4, 0.20 percent dry matter of sulphur (N/S=10 and 0.0625 - 0.125 percent dry matter (DM of phospor (Ca/P=1.0 – 2.0; (2 To stimulate the activity of cellulolytic microbes, its need the fortification of sulphur until reach the optimum level (must be investigated.

  5. Effect of sulfur supplements on cellulolytic rumen micro-organisms and microbial protein synthesis in cattle fed a high fibre diet.

    Science.gov (United States)

    McSweeney, C S; Denman, S E

    2007-11-01

    To examine the effect of sulfur-containing compounds on the growth of anaerobic rumen fungi and the fibrolytic rumen bacteria Ruminococcus albus, Ruminococcus flavefaciens and Fibrobacter succinogenes in pure culture and within the cattle rumen. The effect of two reduced sulfur compounds, 3-mercaptopropionic acid (MPA) or 3-mercapto-1-propanesulfonic acid as the sole S source on growth of pure fibroyltic fungal and bacterial cultures showed that these compounds were capable of sustaining growth. An in vivo trial was then conducted to determine the effect of sulfur supplements (MPA and sodium sulfate) on microbial population dynamics in cattle fed the roughage Dichanthium aristatum. Real-time PCR showed significant increases in fibrolytic bacterial and fungal populations when cattle were supplemented with these compounds. Sulfate supplementation leads to an increase in dry matter intake without a change in whole tract dry matter digestibility. Supplementation of low S-containing diets with either sodium sulfate or MPA stimulates microbial growth with an increase in rumen microbial protein supply to the animal. Through the use of real-time PCR monitoring, a better understanding of the effect of S supplementation on discrete microbial populations within the rumen is provided.

  6. Manipulation of the rumen to increase ruminant production

    International Nuclear Information System (INIS)

    Nolan, J.V.; Leng, R.A.

    1989-01-01

    Manipulation of the rumen should be undertaken with a view to optimizing the supply of specific nutrients to the host animal. Especially important are the volatile fatty acids (glucogenic and non-glucogenic), and dietary and bypass amino acids and lipids. The animal's requirement for these nutrients varies according to its physiological state (growing, pregnant or lactating), its health (disease or parasite load), previous dietary history and also the prevailing climatic conditions. Often the availability of total protein (amino acids) relative to oxidizable substrates (the protein/energy or P/E ratio) is the primary limitation to voluntary feed intake and to efficient use of absorbed nutrients. In these instances, the first objective should be to maximize the yield of microbial amino acids from the rumen by manipulations that increase the net efficiency of microbial synthesis, e.g. by altering methods of feed processing, supplementing with urea and minerals, or eliminating protozoa. Further improvements in the P/E ratio can be attained by supplementing the animal with a dietary protein source with good rumen bypass characteristics in order to provide additional amino acids for intestinal absorption. Other potential manipulations of the rumen include the use of chemicals to modify rumen fermentation patterns and supplementation with long chain fatty acids (LCFA) (as a dense source of energy supplying substrate) in rumen-inert forms (e.g. as Ca-LCFA). However, when energy dense dietary supplements are given, it may be necessary to include a bypass protein source to maintain the P/E ratio. Novel possibilities for manipulation include the use of molecular biology techniques to create microorganisms with enhanced production of enzymes that promote degradation of structural carbohydrates, or with an ability to secrete chemicals that are toxic to protozoa. (author). 68 refs, 4 figs, 1 tab

  7. Comparison of in situ dry matter degradation parameters with in vitro ...

    African Journals Online (AJOL)

    Adem Kamalak

    South African Journal of Animal Science 2005, 35 (4) .... individual means were identified using Tukey's multiple range test (Pearse ..... shown to exert beneficial effects in the form of a reduction of wasteful protein degradation in the rumen.

  8. Ruminal degradation and intestinal digestibility of protein and amino acids in high-protein feedstuffs commonly used in dairy diets.

    Science.gov (United States)

    Paz, H A; Klopfenstein, T J; Hostetler, D; Fernando, S C; Castillo-Lopez, E; Kononoff, P J

    2014-10-01

    A study was conducted to determine the rumen degradation and intestinal digestibility of crude protein (CP) and AA, and AA composition of the rumen-undegradable protein (RUP) from 3 sources of blood meal (BM1, BM2, and BM3), canola meal (CM), low-fat distillers dried grains with solubles (LFDG), soybean meal (SBM), and expeller soybean meal (ESBM). Two Holstein cows fitted with ruminal and proximal duodenal cannulas were used for in situ incubation of 16h and for the mobile bag technique. To correct for bacterial contamination of the RUP, 2 methods were used: purines and DNA as bacterial markers. Ruminal degradations of CP were 85.3, 29.8, 40.7, 75.7, 76.9, 68.8, and 37.0 ± 3.93% for BM1, BM2, BM3, CM, LFDG, SBM, and ESBM, respectively. Ruminal degradation of both total essential AA and nonessential AA followed a similar pattern to that of CP across feedstuffs. Based on the ratio of AA concentration in the RUP to AA concentration in the original feedstuff, ruminal incubation decreased (ratio 1) the concentrations of Ile and Met across feedstuffs. Compared with purines, the use of DNA as bacterial marker resulted in a higher estimate of bacterial CP contamination for CM and lower estimates for LFDG and ESBM. Intestinal digestibility of RUP could not be estimated for BM1, BM3, and SBM due to insufficient recovery of residue. For the remaining feedstuffs, intestinal digestibility of RUP was highest for ESBM, followed by BM2 and LFDG, and lowest for CM: 98.8, 87.9, 89.7, and 72.4 ± 1.40%, respectively. Intestinal absorbable dietary protein was higher for BM2 compared with CM and LFDG, at 61.7, 17.9, and 20.7 ± 2.73% CP, respectively. As prices fluctuate, intestinal absorbable protein or AA may be used as a tool to aid in the selection among feedstuffs with different protein quality. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  9. Rumen microbial genomics

    International Nuclear Information System (INIS)

    Morrison, M.; Nelson, K.E.

    2005-01-01

    Improving microbial degradation of plant cell wall polysaccharides remains one of the highest priority goals for all livestock enterprises, including the cattle herds and draught animals of developing countries. The North American Consortium for Genomics of Fibrolytic Ruminal Bacteria was created to promote the sequencing and comparative analysis of rumen microbial genomes, offering the potential to fully assess the genetic potential in a functional and comparative fashion. It has been found that the Fibrobacter succinogenes genome encodes many more endoglucanases and cellodextrinases than previously isolated, and several new processive endoglucanases have been identified by genome and proteomic analysis of Ruminococcus albus, in addition to a variety of strategies for its adhesion to fibre. The ramifications of acquiring genome sequence data for rumen microorganisms are profound, including the potential to elucidate and overcome the biochemical, ecological or physiological processes that are rate limiting for ruminal fibre degradation. (author)

  10. Feeding different dietary protein to energy ratios to Holstein heifers: effects on growth performance, blood metabolites and rumen fermentation parameters.

    Science.gov (United States)

    Dong, L F; Zhang, W B; Zhang, N F; Tu, Y; Diao, Q Y

    2017-02-01

    Eighteen Chinese Holstein heifers average age 230 ± 14 days were allocated to 1 of 3 dietary crude protein (CP) to metabolizable energy (ME) ratios to examine the effects on growth performance, blood metabolites and rumen fermentation parameters with 90-days experiment. Three different dietary CP:ME ratios were targeted based on the formulation of dietary CP contents of 10.85%, 12.78% and 14.63% on dry matter (DM) basis with similar ME contents (10.42 MJ/kg DM), which were categorized as low, medium and high dietary CP:ME ratios. The actual CP:ME ratios obtained in this study significantly increased from low to high CP:ME ratio groups with a value of 10.59, 11.83 and 13.38 g/MJ respectively. Elevated CP:ME ratios significantly increased CP intake (kg/day) and feed efficiency (FE) which was defined as dry matter intake as a proportion of average daily gain (ADG), whereas little difference was observed in body weight (kg), ADG (kg/day), DM intake (kg/day) and ME intake (MJ/day) among the three different CP:ME ratio groups. Increasing dietary CP to ME ratios significantly increased CP digestibility, whereas digestibility of DM and gross energy remained constant in the current experiment. Blood urea nitrogen and insulin-like growth factor-1 linearly increased with increasing dietary CP:ME ratios. There was significantly dietary treatment effect on rumen fermentation parameters including acetate, propionate, butyrate and total volatile fatty acids. Therefore, this study indicated that increasing dietary CP levels with similar energy content contributed to increased protein intake and its digestibility, as well as FE. Holstein heifers between 200 and 341 kg subjected to 13.38 dietary CP:ME ratio showed improved feed efficiency, nutrient digestibility, some blood metabolites and rumen fermentation characteristics for 0.90 kg/day rate of gain. Journal of Animal Physiology and Animal Nutrition © 2016 Blackwell Verlag GmbH.

  11. Effects of the source of energy and minerals on microbial protein synthesis in rumen using 35S as indicator. Part of a coordinated programme on tracer techniques in studies on the use of non-protein nitrogen in ruminants

    International Nuclear Information System (INIS)

    Durant, M.

    1976-05-01

    Part I. The effect of the nature of carbohydrates and minerals on microbial growth in vitro was studied in vitro to intensify rumen microbial protein synthesis from non-protein nitrogen, using phosphorus incorporation (PR). The nature of starch greatly influences urea utilization. Among the tropical tubers studied, yam Cayenesis and canna Edulis give lower urea utilization than cassava and yam Dumetorum. However, this can be improved by processing. The effects are greatest when the proportion of urea to processed cereal is ca. 4-5%. S-addition as sulfate improves urea utilization with both natural and purified diets. Part II. Results from the Jouy and Ghent laboratories were analyzed statistically, to check the accuracy of the 32 P method for estimating microbial growth, using protein-free substrates. The linear relationship between PR and every other variable was studied. Two multivariate analyses, principal components and multiple regression, were applied. Net NH 3 utilization was predicted using the equations for substrates with proteins likely to be degraded. This method appears more accurate than using the N/P ratio in microflora. Equations should only be used under the specific experiments described

  12. Avaliação de modelos matemáticos para o estudo da cinética de passagem de partículas e de fluidos por bovinos em pastagem recebendo suplementos contendo diferentes níveis de proteína não-degradável no rúmen Evaluation of mathematical models for estimating the kinetics of ruminal Passage of particles and liquid of grazing steers Supplemented with different rumen-undegradable protein levels

    Directory of Open Access Journals (Sweden)

    José Carlos Pereira

    2005-12-01

    Full Text Available Estudaram-se os comportamentos dos parâmetros da cinética de passagem de partículas e de fluidos em bovinos mantidos em pastagem de Brachiaria brizantha, em duas épocas (época 1 = meses de maio, junho e julho/2000; e época 2 = meses de março, abril e maio/2001, recebendo suplementos contendo diferentes níveis de proteína não-degradável no rúmen (PNDR. Na época 1 os animais foram alimentados com suplementos contendo 20, 40 e 60% de PNDR, feno de Brachiaria brizantha e mistura mineral ad libitum, enquanto na época 2, foram distribuídos nos tratamentos: T0 = pastagem de capim-braquiária + mistura mineral ad libitum; T40 = pastagem + suplemento com 40% de PNDR; e T60 = pastagem + suplemento com 60% de PNDR. Na estimativa da taxa de passagem de partículas pelo rúmen, utilizou-se como indicador o Cr-mordente, enquanto a cinética de passagem de fluidos foi estimada com Co-EDTA, durante três períodos, de agosto a setembro/2001, utilizando-se os mesmos animais e tratamentos da época 2. As estimativas dos parâmetros da cinética de passagem de partículas foram determinadas a partir do ajuste dos dados aos modelos bicompartimentais (G1G1, G2G1, G3G1, G4G1, G5G1 e G6G1, enquanto, para a cinética de passagem de fluidos, foi utilizado o modelo G1G1. Os critérios de escolha do melhor modelo foram a freqüência observada dos valores mínimos para o quadrado médio residual, obtido com os ajustes dos diferentes modelos e o número de corridas de sinal dos resíduos padronizados. Os modelos G2G1 e G3G1 mostraram-se mais eficientes na determinação das estimativas dos parâmetros de cinética de passagem de partículas, nas épocas 1 e 2, respectivamente. As estimativas do tempo médio de retenção ruminal de partículas e de fluidos não foram influenciadas pelos diferentes níveis de PNDR do suplemento.The effects of supplements containing different rumen-undegradable protein (RUP levels on the ruminal passage rate of particles and

  13. 菌糠与奶牛常用粗饲料瘤胃降解特性的对比研究%Comparison of Rumen Degradation Characteristics between Spent Mushroom Substrate and Commonly Used Roughages for Dairy Cows

    Institute of Scientific and Technical Information of China (English)

    宫福臣; 韩梅琳; 杨琼; 李杰; 孙晓红

    2013-01-01

    为了系统研究菌糠用作反刍动物饲料的饲用价值,选用3只安装永久性瘤胃瘘管的荷斯坦奶牛为试验动物,采用3×3拉丁方设计,通过尼龙袋法对菌糠、羊草、玉米秸的干物质(DM)、粗蛋白质(CP)、中性洗涤纤维(NDF)和酸性洗涤纤维(ADF)的瘤胃降解率进行测定.粗饲料样品在瘤胃中的培养时间分别为0、6、12、24、36、48、72 h.结果表明:3种粗饲料72 h的DM降解率差异不显著(P>0.05),而其有效降解率由高到低依次为菌糠(29.33%)、羊草(27.41%)、玉米秸(23.39%),且前两者显著高于玉米秸(P<0.05).72 h的CP降解率及其有效降解率以羊草最高,菌糠次之,二者与玉米秸之间差异显著(P<0.05).玉米秸72 h的NDF降解率虽高于羊草和菌糠(P<0.05),但3种粗饲料的有效降解率均较低,由高到低依次为玉米秸(26.12%)、菌糠(25.72%)、羊草(23.73%),无显著差异(P>0.05).而72 h的ADF降解率和有效降解率由高到低也依次为玉米秸(25.36%)、菌糠(24.23%)、羊草(21.83%),变化趋势和差异性与NDF降解规律相一致.由此可见,从3种粗饲料在奶牛瘤胃的消化特性来看,本试验中菌糠的饲用价值与羊草接近,优于玉米秸,因此菌糠作为新型反刍动物粗饲料资源切实可行.%The aim of this study was to evaluate the rumen degradation characteristics of spent mushroom substrate (SMS) as a type of roughage for ruminants.Three Holstein cows with ruminal cannulas were assigned to 3 treatments in a 3 × 3 Latin square design,and the rumen degradation rates of dry matter (DM),crude protein (CP),neutral detergent fiber (NDF) and acid detergent fiber (ADF) of Chinese wildrye (CW),corn stover (CS) and SMS were measured using nylon bag method.The culture time of roughages in the rumen was 0,6,12,24,36,48 and 72 h,respectively.The results showed that there were no differences in DM degradation rate among roughages at 72 h,the effective degradability

  14. Influence of Non-fibrous Carbohydrate and Degradable Intake protein and Ruminal Fermentation ,Nutrien Digestion and performance of Local Sheep

    Directory of Open Access Journals (Sweden)

    Efka AR

    2001-05-01

    Full Text Available The objective of current study was to evaluate the impact dietary non-fibrous carbohydrate ( NFC and ruminally degradable intake protein (DIP concentration have on ruminal fermentation , nutrient digestion and performance of local sheep. The animal had a mean of  liveweight 19.80 ±1.55 kg. four diets ,arranged in a 2x2 factorial ,were formulated to contain either 40 or 50 % NFC and 50 or 60 % of dietary crude protein as DIP .dietary DM contained 25 % Indonesian field grass and 75 % concentrate. Solvent –extracted or formaldehyd  2 % -treated soybean meal were used to alter DIP and corn or soybean hulls to alter NFC level. Percentage of  energy and NDF digestion was similar ( p<0,01 as DIP level decreased in the diets. The soybean hulls was fermentable and total VFA concentration in the rumen increased ( p<0.01, but N-NH3 concentration was decreased ( p<0.01 as DIP level decreased in the diets. Daily live weight gain ( 146.29±25.84 g and body composition ( fat, water , protein and mineral was similar ( p<0.05 among diets. The preponderance ruminal fermentation ,nutrient digestion and performance of local sheeps did not be improved by sincronization of energy and nitrogen release but may more likely be limited by either energy or nitrogen alone. (Animal Production 3(2: 53-61 (2001 Key Word : Carbohydrate, protein, rumen fermentation, nutrients digestion and performance

  15. Rumen Degradation Characteristics and Intestinal Digestibility of Whole Corn Silages from Different Areas%不同产地全株玉米青贮的瘤胃降解特性与小肠消化率的研究

    Institute of Scientific and Technical Information of China (English)

    李洋; 李春雷; 赵洪波; 张永根

    2015-01-01

    本试验旨在研究不同产地全株玉米青贮的营养价值、瘤胃降解特性以及小肠消化率的差异. 选取产自于甘肃( A)、山东( B)、四川( C)、陕西( D)和沈阳( E) 5个地区的全株玉米青贮样品,进行常规营养成分和氨基酸组成分析. 选用3头体重为550 kg左右、安装有永久性瘤胃瘘管的荷斯坦奶牛,采用尼龙袋法测定5 个产地的全株玉米青贮的干物质( DM )、粗蛋白质( CP)、中性洗涤纤维( NDF)、酸性洗涤纤维( ADF)的瘤胃降解参数以及 DM、CP 和氨基酸( AA)的小肠消化率. 结果表明:不同产地的全株玉米青贮常规营养成分存在差异. 5 个产地的全株玉米青贮的DM、CP、NDF、ADF瘤胃有效降解率之间存在差异,其高低顺序分别为B>E>A>D>C、C>A>D>B>E、B>A>D>C>E、D>A>C>B>E. 5个产地的全株玉米青贮的DM、CP和各AA小肠消化率之间存在差异,其中DM、CP和总氨基酸( TAA)小肠消化率的高低顺序分别为C>A>D>B>E、C>A>B>D>E、A>D>B>C>E. 由此得出,不同产地的全株玉米青贮的营养成分含量、瘤胃降解参数以及小肠消化率存在较大差异,在全株玉米青贮的使用中应快速准确地测量其中各种营养成分的含量,以便更有效地利用饲料资源,避免浪费.%The objective of this study was to research the differences in nutritional value, rumen degradation characteristics and intestinal digestibility of whole corn silages from different areas. Five whole corn silage sam?ples were collected from Gansu ( A) , Shandong ( B) , Sichuan ( C) , Shaanxi ( D) and Shenyang ( E) , and the common nutritional components and amino acid composition of feed samples were analysed. Three perma?nent ruminal cannulated Holstein cows with around 550 kg of body weight were used to evaluate rumen degra?dation characteristics of dry matter ( DM) , crude protein ( CP) , neutral detergent fiber ( NDF) and acid deter

  16. In Vitro Biodegradation of Hepatotoxic Indospicine in Indigofera spicata and Its Degradation Derivatives by Camel Foregut and Cattle Rumen Fluids.

    Science.gov (United States)

    Tan, Eddie T T; Al Jassim, Rafat; D'Arcy, Bruce R; Fletcher, Mary T

    2017-08-30

    The known accumulation of the hepatotoxin indospicine in tissues of camels and cattle grazing Indigofera pasture plants is unusual in that free amino acids would normally be expected to be degraded during the fermentation processes in these foregut fermenters. In this study, in vitro experiments were carried out to examine the degradability of indospicine of Indigofera spicata by camel and cattle foregut microbiota. In the first experiment, a 48 h in vitro incubation was carried out using foregut fluid samples that were collected from 15 feral camels and also a fistulated cow. Degradability of indospicine ranged between 97% and 99%, with the higher value of 99% for camels. A pooled sample of foregut fluids from three camels that were on a roughage diet was used in a second experiment to examine the time-dependent degradation of indospicine present in the plant materials. Results indicated that camels' foregut fluids have the ability to biodegrade ∼99% of the indospicine in I. spicata within 48 h of incubation and produced 2-aminopimelamic acid and 2-aminopimelic acid. The time-dependent degradation analysis showed rapid indospicine degradation (65 nmol/h) during the first 8-18 h of incubation followed by a slower degradation rate (12 nmol/h) between 18 and 48 h. Indospicine degradation products were also degraded toward the end of the experiment. The results of these in vitro degradation studies suggest that dietary indospicine may undergo extensive degradation in the foregut of the camel, resulting in trace levels after 48 h. The retention time for plant material in the camel foregut varies depending on feed quality, and the results of this study together with the observed accumulation of indospicine in camel tissues suggest that, although indospicine can be degraded by foregut fermentation, this degradation is not complete before the passage of the digesta into the intestine.

  17. Improving the quality of rice straw by urea and calcium hydroxide on rumen ecology, microbial protein synthesis in beef cattle.

    Science.gov (United States)

    Polyorach, S; Wanapat, M

    2015-06-01

    Four rumen-fistulated beef cattle were randomly assigned to four treatments according to a 4 × 4 Latin square design to study the influence of urea and calcium hydroxide [Ca(OH)2 ] treatment of rice straw to improve the nutritive value of rice straw. Four dietary treatments were as follows: untreated rice straw, 50 g/kg urea-treated rice straw, 20 g/kg urea + 20 g/kg calcium hydroxide-treated rice straw and 30 g/kg urea + 20 g/kg calcium hydroxide-treated rice straw. All animals were kept in individual pens and fed with concentrate at 0.5 g/kg of BW (DM), rice straw was fed ad libitum. The experiment was conducted for four periods, and each period lasted for 21 days. During the first 14 days, DM feed intake measurements were made while during the last 7 days, all cattle were moved to metabolism crates for total faeces and urine collections. The results revealed that 20 g/kg urea + 20 g/kg calcium hydroxide-treated rice straw improved the nutritive value of rice straw, in terms of dry matter intake, digestibility, ruminal volatile fatty acids, population of bacteria and fungi, nitrogen retention and microbial protein synthesis. Based on this study, it could be concluded that using urea plus calcium hydroxide was one alternative method to improve the nutritive value of rice straw, rumen ecology and fermentation and thus a reduction of treatment cost. Journal of Animal Physiology and Animal Nutrition © 2014 Blackwell Verlag GmbH.

  18. Rumen microbial growth estimation using in vitro radiophosphorous incorporation technique

    International Nuclear Information System (INIS)

    Bueno, Ives Claudio da Silva; Machado, Mariana de Carvalho; Cabral Filho, Sergio Lucio Salomon; Gobbo, Sarita Priscila; Vitti, Dorinha Miriam Silber Schmidt; Abdalla, Adibe Luiz

    2002-01-01

    Rumen microorganisms are able to transform low biological value nitrogen of feed stuff into high quality protein. To determine how much microbial protein that process forms, radiomarkers can be used. Radiophosphorous has been used to mark microbial protein, as element P is present in all rumen microorganisms (as phospholipids) and the P:N ratio of rumen biomass is quite constant. The aim of this work was to estimate microbial synthesis from feedstuff commonly used in ruminant nutrition in Brazil. Tested feeds were fresh alfalfa, raw sugarcane bagasse, rice hulls, rice meal, soybean meal, wheat meal, Tifton hay, leucaena, dehydrated citrus pulp, wet brewers' grains and cottonseed meal. 32 P-labelled phosphate solution was used as marker for microbial protein. Results showed the diversity of feeds by distinct quantities of nitrogen incorporated into microbial mass. Low nutrient availability feeds (sugarcane bagasse and rice hulls) promoted the lowest values of incorporated nitrogen. Nitrogen incorporation showed positive relationship (r=0.56; P=0.06) with the rate of degradation and negative relationship (r=-0.59; P<0.05) with fiber content of feeds. The results highlight that easier fermentable feeds (higher rates of degradation) and/or with lower fiber contents promote a more efficient microbial growth and better performance for the host animal. (author)

  19. Rumen microbial growth estimation using in vitro radiophosphorous incorporation technique

    Energy Technology Data Exchange (ETDEWEB)

    Bueno, Ives Claudio da Silva; Machado, Mariana de Carvalho; Cabral Filho, Sergio Lucio Salomon; Gobbo, Sarita Priscila; Vitti, Dorinha Miriam Silber Schmidt; Abdalla, Adibe Luiz [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil)

    2002-07-01

    Rumen microorganisms are able to transform low biological value nitrogen of feed stuff into high quality protein. To determine how much microbial protein that process forms, radiomarkers can be used. Radiophosphorous has been used to mark microbial protein, as element P is present in all rumen microorganisms (as phospholipids) and the P:N ratio of rumen biomass is quite constant. The aim of this work was to estimate microbial synthesis from feedstuff commonly used in ruminant nutrition in Brazil. Tested feeds were fresh alfalfa, raw sugarcane bagasse, rice hulls, rice meal, soybean meal, wheat meal, Tifton hay, leucaena, dehydrated citrus pulp, wet brewers' grains and cottonseed meal. {sup 32} P-labelled phosphate solution was used as marker for microbial protein. Results showed the diversity of feeds by distinct quantities of nitrogen incorporated into microbial mass. Low nutrient availability feeds (sugarcane bagasse and rice hulls) promoted the lowest values of incorporated nitrogen. Nitrogen incorporation showed positive relationship (r=0.56; P=0.06) with the rate of degradation and negative relationship (r=-0.59; P<0.05) with fiber content of feeds. The results highlight that easier fermentable feeds (higher rates of degradation) and/or with lower fiber contents promote a more efficient microbial growth and better performance for the host animal. (author)

  20. Metabolizable protein systems in ruminant nutrition: A review

    Directory of Open Access Journals (Sweden)

    Lalatendu Keshary Das

    2014-08-01

    Full Text Available Protein available to ruminants is supplied by both microbial and dietary sources. Metabolizable protein (MP is the true protein which is absorbed by the intestine and supplied by both microbial protein and protein which escapes degradation in the rumen; the protein which is available to the animal for maintenance, growth, fetal growth during gestation, and milk production. Thus, the concept of balancing ruminant rations basing on only dietary crude protein (CP content seems erroneous. In India, ruminant rations are still balanced for digestible CP and total digestible nutrients for protein and energy requirements, respectively. Traditional feed analysis methods such as proximate analysis and detergent analysis consider feed protein as a single unit and do not take into account of the degradation processes that occur in rumen and passage rates of feed fractions from rumen to intestine. Therefore, the protein requirement of ruminants should include not only the dietary protein source, but also the microbial CP from rumen. The MP systems consider both the factors, thus predict the protein availability more accurately and precisely. This system is aptly designed to represent the extent of protein degradation in the rumen and the synthesis of microbial protein as variable functions. Feed protein fractions, i.e., rumen degradable protein and rumen undegradable protein play vital roles in meeting protein requirements of rumen microbes and host animal, respectively. With the advent of sophisticated nutrition models such as Cornell net carbohydrate and protein system, National Research Council, Agricultural Research Council, Cornell Penn Miner Dairy and Amino Cow; ration formulation has moved from balancing diets from CP to MP, a concept that describes the protein requirements of ruminantsat intestinal level, and which is available to animals for useful purposes.

  1. Intensive liquid feeding of dairy calves with a medium crude protein milk replacer: Effects on performance, rumen, and blood parameters.

    Science.gov (United States)

    de Paula, M R; Oltramari, C E; Silva, J T; Gallo, M P C; Mourão, G B; Bittar, C M M

    2017-06-01

    The objective of this study was to evaluate the effects of different liquid-feeding systems using a medium crude protein milk replacer on performance, rumen, and blood parameters. Thirty newborn Holstein calves were blocked according to birth weight and date of birth, and randomly distributed to different liquid-feeding systems: conventional (4 L/d), intensive (8 L/d), or step-up/step-down (wk 1, 4 L/d; wk 2 to 6, 8 L/d; wk 7 and 8, 4 L/d). The commercial milk replacer (12.5% solids, 20.2% crude protein, 15.6% fat) was fed twice daily (0700 and 1700 h) until calves were weaned, at 8 wk of age. Calves were individually housed in wood hutches, with free access to water and starter concentrate, and to hay only after weaning. They were followed through 10 wk of age. Milk replacer and starter intake were inversely affected by feeding system. After weaning, starter intake and hay intake were similar among feeding systems. Total dry matter intake was higher during the liquid-feeding period for calves on the intensive system compared to calves on the conventional system, but conventional feeding resulted in the highest dry matter intake after weaning. Feed efficiency was similar among feeding systems before and after weaning. Average body weight and daily gain were not affected by feeding system before or after weaning. During liquid feeding, diarrhea occurrence was lower for calves on the conventional system; however, when calves on the step-up/step-down system were fed lower volumes of liquid feed, diarrhea occurrence was similar to that of calves on the conventional system. Plasma concentrations of β-hydroxybutyrate were higher for calves on the conventional system, reflecting starter intake. Rumen pH, short-chain fatty acids, and N-NH 3 were not affected by feeding system. Feeding higher volumes of milk replacer with a medium crude protein content had no beneficial effect on the performance of calves up to 10 wk of age. Copyright © 2017 American Dairy Science

  2. Anti-nutritional Factors and Ruminal Dry Matter and Crude Protein Degradability of Gamma and Microwave Irradiated Native Rapeseed

    Directory of Open Access Journals (Sweden)

    sayyed roohollah ebrahimimahmoudabad

    2016-08-01

    Full Text Available Introduction Native rapeseed (NRS is planted in some parts of Iran because of climatic condition. The consumption of NRS in animal nutrition is limited by anti-nutritional such as phytic acid and glucosinolate. Moreover, the protein of NRS is highly degraded by rumen microorganisms. Several processing methods have been used to enhance the nutritive value of whole oilseeds, including extrusion, roasting, toasting and Jet-Sploding. However, most heat processing methods adversely affect protein digestibility in the small intestine. Recently, other processing methods such as processing by gamma and microwave irradiation have been noticed. Therefore, this research was carried out to evaluate the effects of gamma irradiation (15, 30 and 45 kGy and microwave irradiation (800 W for 2, 4 and 6 min on ruminal dry matter (DM and crude protein (CP degradability, in vitro CP digestibility, anti-nutritional factors (glucosinolate and phytic acid and chemical composition of NRS. Materials and Methods Chemical composition (DM, CP, EE and Ash of untreated and irradiated NRS was determined by AOAC methods. Then, sufficient water was added to the sample to increase the moisture content to 250 g/kg. Gamma irradiation was completed by using a cobalt-60 irradiator at 20 ºC. The dose rate determined by Fricke dosimetry was 0.36 Gy/s. Another three samples (500 g each were subjected to microwave irradiation at a power of 800 W for 2, 4 and 6 min. Phytic acid and glucosinolate contents of untreated and irradiated samples were determined by standard methods. Degradation kinetics of DM or CP were determined according to in situ procedure. Six grams of untreated or irradiated NRS were incubated in the rumen of three ruminally fistulated Taleshi bulls for 0, 2, 4, 8, 16, 24 and 48 h. Bags were placed in the rumen just before the bulls were offered their first meal. After retrieval from the rumen, bags were thoroughly washed with tap water until the rinsing water was clear

  3. Effect of salseed-meal tannins on protein synthesis, /sup 35/S incorporation and cellulose digestibility by rumen microbes in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Singh, K; Arora, S P [Haryana Agricultural Univ., Hissar (India)

    1980-10-01

    Tannins from seed-meal of sal (Shorea robusta Gaertn. F.) were fractionated after treatments into ethyl acetate (EA) and lead acetate (LA) fractions. In trial 1, incorporation of /sup 35/S from (NH/sub 4/)/sub 2//sup 35/SO/sub 4/ into microbial protein declined due to the effect of both 2% EA and LA fractions as compared to control. Microbial protein synthesis was depressed significantly (P < 0.01) in treatments containing 1.0 and 2.0% of EA fractions, and 0.5, 1.0 and 2.0% of LA fractions. The microbial cellulolytic activity also exhibited a similar trend in experiment 2. In experiment 3, in which all parameters were studied simultaneously, a significant depression was found in the values of microbial protein synthesis, /sup 35/S incorporation and cellulolysis at all levels of both the tannin fractions. It may be inferred that both the tannin fractions from salseed-meal showed antimicrobial activity and LA fraction seems to be more deleterious than EA fraction for rumen metabolism. The experiments were conducted in vitro using rumen liquor of a non-pregnant dry cow having permanent rumen fistula.

  4. Effect of inclusion of different levels of silage on rumen microbial population and microbial protein synthesis in dairy steers fed on rice straw

    Directory of Open Access Journals (Sweden)

    Thien Truong Giang Nguyen

    2017-02-01

    Full Text Available Objective Leucaena leucocephala (Leucaena is a perennial tropical legume that can be directly grazed or harvested and offered to ruminants as hay, silage, or fresh. However, Leucaena contain phenolic compounds, which are considered anti-nutritional factors as these may reduce intake, digestibility and thus animal performance. Therefore, the objective of this experiment was to determine effects of Leucaena silage (LS feeding levels on rumen microbial populations, N-balance and microbial protein synthesis in dairy steers. Methods Four, rumen fistulated dairy steers with initial weight of 167±12 kg were randomly assigned to receive dietary treatments according to a 4×4 Latin square design. Treatments were as followings: T1 = untreated rice straw (RS; Control, T2 = 70% RS+30% LS, T3 = 40% RS+60% LS, and T4 = 100% LS. Dairy steers were fed rice straw and LS ad libitum and supplemented with concentrate at 0.2% of body weight/d. Results Results revealed that the rumen microbial population, especially cellulolytic, proteolytic bacteria and fungal zoospores were enhanced in steers that received 60% of LS (p0.05. Protozoal population was linearly decreased with increasing level of LS (p<0.05. Moreover, N-balance and microbial protein synthesis were enhanced by LS feeding (p<0.05 and were the highest in 60% LS group. Conclusion Based on this study, it could be concluded that replacement of RS with 60% LS significantly improved microbial population and microbial protein synthesis in diary steers.

  5. Chemical composition and ruminal degradation kinetics of crude protein and amino acids, and intestinal digestibility of amino acids from tropical forages

    Directory of Open Access Journals (Sweden)

    Lidia Ferreira Miranda

    2012-03-01

    Full Text Available The objective of this research was to determine the chemical composition and ruminal degradation of the crude protein (CP, total and individual amino acids of leaves from tropical forages: perennial soybean (Neonotonia wightii, cassava (Manihot esculenta, leucaena (Leucaena leucocephala and ramie (Boehmeria nivea, and to estimate the intestinal digestibility of the rumen undegradable protein (RUDP and individual amino acids of leaves from the tropical forages above cited, but including pigeon pea (Cajanus cajan. Three nonlactating Holstein cows were used to determine the in situ ruminal degradability of protein and amino acids from leaves (6, 18 and 48 hours of ruminal incubation. For determination of the intestinal digestibility of RUDP, the residue from ruminal incubation of the materials was used for 18 hours. A larger concentration of total amino acids for ramie and smaller for perennial soybean were observed; however, they were very similar in leucaena and cassava. Leucine was the essential amino acid of greater concentration, with the exception of cassava, which exhibited a leucine concentration 40.45% smaller. Ramie showed 14.35 and 22.31% more lysine and methionine, respectively. The intestinal digestibility of RUDP varied from 23.56; 47.87; 23.48; 25.69 and 10.86% for leucaena, perennial soybean, cassava, ramie and pigeon pea, respectively. The individual amino acids of tropical forage disappeared in different extensions in the rumen. For the correct evaluation of those forages, one should consider their composition of amino acids, degradations and intestinal digestibility, once the amino acid composition of the forage does not reflect the amino acid profiles that arrived in the small intestine. Differences between the degradation curves of CP and amino acids indicate that degradation of amino acids cannot be estimated through the degradation curve of CP, and that amino acids are not degraded in a similar degradation profile.

  6. A 100-Year Review: Protein and amino acid nutrition in dairy cows.

    Science.gov (United States)

    Schwab, Charles G; Broderick, Glen A

    2017-12-01

    Considerable progress has been made in understanding the protein and amino acid (AA) nutrition of dairy cows. The chemistry of feed crude protein (CP) appears to be well understood, as is the mechanism of ruminal protein degradation by rumen bacteria and protozoa. It has been shown that ammonia released from AA degradation in the rumen is used for bacterial protein formation and that urea can be a useful N supplement when lower protein diets are fed. It is now well documented that adequate rumen ammonia levels must be maintained for maximal synthesis of microbial protein and that a deficiency of rumen-degradable protein can decrease microbial protein synthesis, fiber digestibility, and feed intake. Rumen-synthesized microbial protein accounts for most of the CP flowing to the small intestine and is considered a high-quality protein for dairy cows because of apparent high digestibility and good AA composition. Much attention has been given to evaluating different methods to quantify ruminal protein degradation and escape and for measuring ruminal outflows of microbial protein and rumen-undegraded feed protein. The methods and accompanying results are used to determine the nutritional value of protein supplements and to develop nutritional models and evaluate their predictive ability. Lysine, methionine, and histidine have been identified most often as the most-limiting amino acids, with rumen-protected forms of lysine and methionine available for ration supplementation. Guidelines for protein feeding have evolved from simple feeding standards for dietary CP to more complex nutrition models that are designed to predict supplies and requirements for rumen ammonia and peptides and intestinally absorbable AA. The industry awaits more robust and mechanistic models for predicting supplies and requirements of rumen-available N and absorbed AA. Such models will be useful in allowing for feeding lower protein diets and increased efficiency of microbial protein synthesis

  7. Postruminal degradation of crude protein, neutral detergent fibre and starch of maize and grass silages in dairy cows

    DEFF Research Database (Denmark)

    Ali, M.; Weisbjerg, Martin Riis; Cone, J.W.

    2012-01-01

    samples of grass silage were used. The samples were selected to represent a broad range in digestibility and chemical composition. Prior to the intestinal incubations, samples were incubated in the rumen for 6 h (starch), 12 h (CP) or 24 h (aNDFom) using the rumen nylon bag technique. Residues from....... The objective of this study was to develop a unique dataset on the ruminal degradability and the postruminal digestibility of CP, NDF (aNDFom, amylase neutral detergent fibre organic matter basis) and starch in maize and grass silages, using the mobile nylon bag technique. Twenty samples of maize silage and 20...... in the maize and grass silages. The results proved the assumption of the Dutch feed evaluation system that the rumen undegraded starch is completely digested in the small intestine of dairy cows. Regression showed that the rumen degradability, the intestinal digestibility and the total tract undigested...

  8. Invited review: Essential oils as modifiers of rumen microbial fermentation.

    Science.gov (United States)

    Calsamiglia, S; Busquet, M; Cardozo, P W; Castillejos, L; Ferret, A

    2007-06-01

    Microorganisms in the rumen degrade nutrients to produce volatile fatty acids and synthesize microbial protein as an energy and protein supply for the ruminant, respectively. However, this fermentation process has energy (losses of methane) and protein (losses of ammonia N) inefficiencies that may limit production performance and contribute to the release of pollutants to the environment. Antibiotic ionophores have been very successful in reducing these energy and protein losses in the rumen, but the use of antibiotics in animal feeds is facing reduced social acceptance, and their use has been banned in the European Union since January 2006. For this reason, scientists have become interested in evaluating other alternatives to control specific microbial populations to modulate rumen fermentation. Essential oils can interact with microbial cell membranes and inhibit the growth of some gram-positive and gram-negative bacteria. As a result of such inhibition, the addition of some plant extracts to the rumen results in an inhibition of deamination and methanogenesis, resulting in lower ammonia N, methane, and acetate, and in higher propionate and butyrate concentrations. Results have indicated that garlic oil, cinnamaldehyde (the main active component of cinnamon oil), eugenol (the main active component of the clove bud), capsaicin (the active component of hot peppers), and anise oil, among others, may increase propionate production, reduce acetate or methane production, and modify proteolysis, peptidolysis, or deamination in the rumen. However, the effects of some of these essential oils are pH and diet dependent, and their use may be beneficial only under specific conditions and production systems. For example, capsaicin appears to have small effects in high-forage diets, whereas the changes observed in high-concentrate diets (increases in dry matter intake and total VFA, and reduction in the acetateto-propionate ratio and ammonia N concentration) may be beneficial

  9. Technical note: Protozoa-specific antibodies raised in sheep plasma bind to their target protozoa in the rumen.

    Science.gov (United States)

    Williams, Y J; Rea, S M; Popovski, S; Skillman, L C; Wright, A-D G

    2014-12-01

    Binding of IgG antibodies to Entodinium spp. in the rumen of sheep (Ovis aries) was investigated by adding IgG, purified from plasma, directly into the rumen. Plasma IgG was sourced from sheep that had or had not been immunized with a vaccine containing whole fixed Entodinium spp. cells. Ruminal fluid was sampled approximately 2 h after each antibody dosing. Binding of protozoa by a specific antibody was detected using an indirect fluorescent antibody test. An antibody titer in the ruminal fluid was determined by ELISA, and the concentration of ruminal fluid ammonia-N and ruminal pH were also determined. Entodinium spp. and total protozoa from IgG-infused sheep were enumerated by microscopic counts. Two-hourly additions of IgG maintained a low antibody titer in the rumen for 12 h and the binding of the antibody to the rumen protozoa was demonstrated. Increased ammonia-N concentrations and altered ruminal fluid pH patterns indicated that additional fermentation of protein was occurring in the rumen after addition of IgG. No reduction in numbers of Entodinium spp. was observed (P>0.05). Although binding of antibodies to protozoa has been demonstrated in the rumen, it is unclear how much cell death occurred. On the balance of probability, it would appear that the antibody was degraded or partially degraded, and the impact of this on protozoal populations and the measurement of a specific titer is also unclear.

  10. The establishment of a protein degradability data base for dairy ...

    African Journals Online (AJOL)

    The degradability values ranged from 96,2; 91,9 and 88,7% for ... McDonald, 1979; Kristensen, et ai., 1982; Lindberg,. 1983 ... mass of 570 kg, and average production of 5000 kg ... Dicalcium phosphate'. 0,5. Commercial vitamin and trace mineral premix. 0,3 .... effects on the degradation of most protein supplements.

  11. Cellular proteostasis: degradation of misfolded proteins by lysosomes

    Science.gov (United States)

    Jackson, Matthew P.

    2016-01-01

    Proteostasis refers to the regulation of the cellular concentration, folding, interactions and localization of each of the proteins that comprise the proteome. One essential element of proteostasis is the disposal of misfolded proteins by the cellular pathways of protein degradation. Lysosomes are an important site for the degradation of misfolded proteins, which are trafficked to this organelle by the pathways of macroautophagy, chaperone-mediated autophagy and endocytosis. Conversely, amyloid diseases represent a failure in proteostasis, in which proteins misfold, forming amyloid deposits that are not degraded effectively by cells. Amyloid may then exacerbate this failure by disrupting autophagy and lysosomal proteolysis. However, targeting the pathways that regulate autophagy and the biogenesis of lysosomes may present approaches that can rescue cells from the deleterious effects of amyloidogenic proteins. PMID:27744333

  12. In vitro microbial protein synthesis, ruminal degradation and post-ruminal digestibility of crude protein of dairy rations containing Quebracho tannin extract.

    Science.gov (United States)

    Castro-Montoya, J; Westreicher-Kristen, E; Henke, A; Diaby, M; Susenbeth, A; Dickhoefer, U

    2018-02-01

    This study evaluated the effects of Quebracho tannin extract (QTE) on in vitro ruminal fermentation, chemical composition of rumen microbes, ruminal degradation and intestinal digestibility of crude protein (iCPd). Three treatments were tested, the control (basal diet without QTE), the basal diet with 15 g QTE/kg dry matter (DM) and the basal diet with 30 g QTE/kg DM. The basal diet contained (g/kg DM): 339 grass silage, 317 maize silage and 344 concentrate. In vitro gas production kinetic was determined using the Hohenheim gas test (Experiment 1). The Ankom RF technique, a batch system with automatic gas pressure recordings, was used to determine in vitro production of short-chain fatty acids (SCFA) and ammonia-nitrogen concentration (NH 3 -N), as well as nitrogen and purine bases content in liquid-associated microbes (LAM) and in a residue of undegraded feed and solid-associated microbes (Feed+SAM) (Experiment 2). Ruminal degradation and iCPd were determined using the nylon bag technique and the mobile nylon bag technique, respectively (Experiment 3). Gas production (Experiment 1), total SCFA and NH 3 -N (Experiment 2) decreased with increasing QTE levels. Microbial mass and composition of LAM were not affected by QTE, but total mass of Feed+SAM linearly increased, likely due to decreased substrate degradation with increasing QTE levels. The total amount of N in microbial mass and undegraded feed after the in vitro incubation increased with increasing QTE levels, suggesting a potential greater N flow from the rumen to the duodenum. In contrast to in vivo studies with the same QTE, no effects were detected on ruminal effective degradability and iCPd, when using the nylon bag techniques. Based on the in vitro procedures, QTE increased the supply of N post-rumen; however, some evidence of a decreased fibre degradation were also observed. Therefore, the benefit of adding QTE to diets of cattle is still questionable. © 2017 Blackwell Verlag GmbH.

  13. Dissection of membrane protein degradation mechanisms by reversible inhibitors

    International Nuclear Information System (INIS)

    Hare, J.F.

    1988-01-01

    The degradation of slowly turning over 125I-lactoperoxidase-labeled plasma membrane polypeptides in response to reversible temperature and lysosomotropic inhibitors was studied in rat hepatoma cultures. Cells were radiolabeled and left for 24 h to allow the removal of rapidly degraded proteins. Remaining trichloroacetic acid-precipitable protein was degraded (t 1/2 = 40-68 h) by an apparent first order process 60-86% sensitive to 10 mM NH4Cl or 5 mM methylamine and greater than 95% inhibited by temperature reduction to 18 degrees C. Thus, membrane proteins are selected for degradation in a time-dependent manner by a system which is sensitive to both 18 degrees C and to lysosomotropic amines. When inhibitory conditions were removed after 40-48 h, degradation of 125I-labeled protein resumed at the same rate as that seen in their absence. Since membrane proteins do not exhibit accelerated degradation after removal of inhibitory conditions, there can be no marking or sorting of those proteins destined for degradation during the 40-h exposure to inhibitory conditions. Exposure to amines or 18 degrees C did not affect the position of two-dimensionally resolved labeled polypeptides. Fractionation of labeled cells on Percoll gradients after 40 h of exposure to low temperature or amines showed that labeled protein remained in the plasma membrane fractions of the gradient although shifted to a slightly lower buoyant density in the presence of amines. These results support the notion that selection of plasma membrane proteins for degradation requires their internalization into acidic vesicles. Lysosomotropic amines and reduced temperature interfere with the selection process by preventing membrane fusion events

  14. Proteasomal and Lysosomal Protein Degradation and Heart Disease

    OpenAIRE

    Wang, Xuejun; Robbins, Jeffrey

    2013-01-01

    In the cell, the proteasome and lysosomes represent the most important proteolytic machineries, responsible for the protein degradation in the ubiquitin-proteasome system (UPS) and autophagy, respectively. Both the UPS and autophagy are essential to protein quality and quantity control. Alterations in cardiac proteasomal and lysosomal degradation are remarkably associated with most heart disease in humans and are implicated in the pathogenesis of congestive heart failure. Studies carried out ...

  15. Knowns and unknowns of plasma membrane protein degradation in plants.

    Science.gov (United States)

    Liu, Chuanliang; Shen, Wenjin; Yang, Chao; Zeng, Lizhang; Gao, Caiji

    2018-07-01

    Plasma membrane (PM) not only creates a physical barrier to enclose the intracellular compartments but also mediates the direct communication between plants and the ever-changing environment. A tight control of PM protein homeostasis by selective degradation is thus crucial for proper plant development and plant-environment interactions. Accumulated evidences have shown that a number of plant PM proteins undergo clathrin-dependent or membrane microdomain-associated endocytic routes to vacuole for degradation in a cargo-ubiquitination dependent or independent manner. Besides, several trans-acting determinants involved in the regulation of endocytosis, recycling and multivesicular body-mediated vacuolar sorting have been identified in plants. More interestingly, recent findings have uncovered the participation of selective autophagy in PM protein turnover in plants. Although great progresses have been made to identify the PM proteins that undergo dynamic changes in subcellular localizations and to explore the factors that control the membrane protein trafficking, several questions remain to be answered regarding the molecular mechanisms of PM protein degradation in plants. In this short review article, we briefly summarize recent progress in our understanding of the internalization, sorting and degradation of plant PM proteins. More specifically, we focus on discussing the elusive aspects underlying the pathways of PM protein degradation in plants. Copyright © 2018 Elsevier B.V. All rights reserved.

  16. The effect of rumen degradable protein level and source on the ...

    African Journals Online (AJOL)

    Histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, valine and total essential AA concentrations were not affected by dietary RDP level. Increasing the urea-N content of RDP supplements significantly decreased isoleucine, leucine, lysine, phenylalanine, tryptophan, valine and total essential amino ...

  17. Protein degradation pathways in Parkinson's disease: curse or blessing.

    Science.gov (United States)

    Ebrahimi-Fakhari, Darius; Wahlster, Lara; McLean, Pamela J

    2012-08-01

    Protein misfolding, aggregation and deposition are common disease mechanisms in many neurodegenerative diseases including Parkinson's disease (PD). Accumulation of damaged or abnormally modified proteins may lead to perturbed cellular function and eventually to cell death. Thus, neurons rely on elaborated pathways of protein quality control and removal to maintain intracellular protein homeostasis. Molecular chaperones, the ubiquitin-proteasome system (UPS) and the autophagy-lysosomal pathway (ALP) are critical pathways that mediate the refolding or removal of abnormal proteins. The successive failure of these protein degradation pathways, as a cause or consequence of early pathological alterations in vulnerable neurons at risk, may present a key step in the pathological cascade that leads to spreading neurodegeneration. A growing number of studies in disease models and patients have implicated dysfunction of the UPS and ALP in the pathogenesis of Parkinson's disease and related disorders. Deciphering the exact mechanism by which the different proteolytic systems contribute to the elimination of pathogenic proteins, like α-synuclein, is therefore of paramount importance. We herein review the role of protein degradation pathways in Parkinson's disease and elaborate on the different contributions of the UPS and the ALP to the clearance of altered proteins. We examine the interplay between different degradation pathways and provide a model for the role of the UPS and ALP in the evolution and progression of α-synuclein pathology. With regards to exciting recent studies we also discuss the putative potential of using protein degradation pathways as novel therapeutic targets in Parkinson's disease.

  18. Estimasi Sintesis Protein Mikrobia Rumen Menggunakan Ekskresi Derivat Purin dalam Urin dengan Teknik Spot Sampling pada Kambing Bligon dan Kambing Kejobong

    Directory of Open Access Journals (Sweden)

    Dianestu Putra

    2016-11-01

    Full Text Available This study were aimed to determine the correlation between concentration of purine derivatives (PD in spot sample with PD total excretion in Bligon and Kejobong goats and determine the appropriate sampling time, in order to predicting microbial protein synthesis in both breeds. Six male Bligon goats and six male Kejobong goats with age range from 8 to 14 months and body weight from 16 to 21 kg were placed in metabolism cages. Peanut straw and water were given to both groups of goats through ad libitum feeding and drinking. The study was done in 14 days for adaptation, 3 days for collection. Sample of feeds, feed residues, and feces were collected and then analyzed to determine dry matter and organic matter content. Spot urine and the total daily urine samples were also collected. Samples collection of spot sampling technique was run by taking the urine periodically with 3 hours intervals at 24 hours. Urine samples were analyzed for the content of creatinine and PD which includes allantoin, uric acid, xanthine, and hypoxanthine. Data were tested for the correlation between concentration of PD spot urine sample with total PD daily excretion. It is known that the concentration of PD and creatinine (µmol/L for Bligon were 1,418.40 and 202.85 respectively, while for Kejobong were 1,547.40 and 219.68 respectively. Total excretion of PD, allantoin, uric acid, xanthyne and hypoxanthine and creatinine (µmol/W0,75/day for Bligon were 114.14, 95.86, 17.31, 0.97, and 16.40 respectively, with microbial protein synthesis efficiency was 4.61 g N/kg degraded of organic matter in rumen (DOMR. Total excretion of PD allantoin, uric acid, xanthyne and hypoxanthine and creatinine (µmol/W0,75/day for Kejobong were 180.18, 158.17, 20.60, 1.40, and 24.87 respectively, with microbial protein synthesis efficiency was 6.90 g N/kg DOMR. Based on this study also known that the best time for spot sampling to determine the total excretion of PD in Bligon was in the range

  19. Herp enhances ER-associated protein degradation by recruiting ubiquilins

    International Nuclear Information System (INIS)

    Kim, Tae-Yeon; Kim, Eunmin; Yoon, Sungjoo Kim; Yoon, Jong-Bok

    2008-01-01

    ER-associated protein degradation (ERAD) is a protein quality control system of ER, which eliminates misfolded proteins by proteasome-dependent degradation and ensures export of only properly folded proteins from ER. Herp, an ER membrane protein upregulated by ER stress, is implicated in regulation of ERAD. In the present study, we show that Herp interacts with members of the ubiquilin family, which function as a shuttle factor to deliver ubiquitinated substrates to the proteasome for degradation. Knockdown of ubiquilin expression by small interfering RNA stabilized the ERAD substrate CD3δ, whereas it did not alter or increased degradation of non-ERAD substrates tested. CD3δ was stabilized by overexpressed Herp mutants which were capable of binding to ubiquilins but were impaired in ER membrane targeting by deletion of the transmembrane domain. Our data suggest that Herp binding to ubiquilin proteins plays an important role in the ERAD pathway and that ubiquilins are specifically involved in degradation of only a subset of ubiquitinated targets, including Herp-dependent ERAD substrates

  20. Protein oxidation and degradation caused by particulate matter

    Science.gov (United States)

    Lai, Ching-Huang; Lee, Chun-Nin; Bai, Kuan-Jen; Yang, You-Lan; Chuang, Kai-Jen; Wu, Sheng-Ming; Chuang, Hsiao-Chi

    2016-09-01

    Particulate matter (PM) modulates the expression of autophagy; however, the role of selective autophagy by PM remains unclear. The objective of this study was to determine the underlying mechanisms in protein oxidation and degradation caused by PM. Human epithelial A549 cells were exposed to diesel exhaust particles (DEPs), urban dust (UD), and carbon black (CB; control particles). Cell survival and proliferation were significantly reduced by DEPs and UD in A549 cells. First, benzo(a)pyrene diolepoxide (BPDE) protein adduct was caused by DEPs at 150 μg/ml. Methionine oxidation (MetO) of human albumin proteins was induced by DEPs, UD, and CB; however, the protein repair mechanism that converts MetO back to methionine by methionine sulfoxide reductases A (MSRA) and B3 (MSRB3) was activated by DEPs and inhibited by UD, suggesting that oxidized protein was accumulating in cells. As to the degradation of oxidized proteins, proteasome and autophagy activation was induced by CB with ubiquitin accumulation, whereas proteasome and autophagy activation was induced by DEPs without ubiquitin accumulation. The results suggest that CB-induced protein degradation may be via an ubiquitin-dependent autophagy pathway, whereas DEP-induced protein degradation may be via an ubiquitin-independent autophagy pathway. A distinct proteotoxic effect may depend on the physicochemistry of PM.

  1. The establishment of a protein degradability data base for dairy ...

    African Journals Online (AJOL)

    The establishment of a protein degradability data base for dairy cattle using the nylon bag technique. 1. Protein sources. LJ Erasmus, J Prinsloo, HH Meissner. Abstract. No Abstract. Full Text: EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT · AJOL African Journals ...

  2. Stage of lactation and corresponding diets affect in situ protein degradation by dairy cows.

    Science.gov (United States)

    Schadt, I; Mertens, D R; Van Soest, P J; Azzaro, G; Licitra, G

    2014-12-01

    The influence of stage of lactation and corresponding diets on rates of protein degradation (kd) is largely unstudied. Study objectives were to measure and compare in situ ruminal kd of crude protein (CP) and estimate rumen CP escape (rumen-undegradable protein; RUP) of selected feeds by cows at 3 stages of lactation fed corresponding diets, and to determine the incubation times needed in an enzymatic in vitro procedure, using 0.2 units of Streptomyces griseus protease per percent of true CP, that predicted in situ RUP. Residue CP was measured after in situ fermentation for 4, 8, 12, 24, 36, 48, and 72 h of 5 protein sources and 3 total mixed rations, which were fed to the in situ cows. Two nonlactating (dry) cows and 2 cows each at 190 (mid) and 90 (peak) days of lactation were used. Each pair of cows was offered free-choice diets that differed in composition to meet their corresponding nutrient requirements. Diets had decreasing proportions of forages and contained (dry matter basis) 11.9, 15.1 and 16.4% CP and 54.3, 40.3 and 35.3% neutral detergent fiber, for dry, mid, and peak TMR (TMR1, TMR2, and TMR3), respectively. Intakes were 10.3, 21.4, and 23.8kg of dry matter/d, respectively. Kinetic CP fractions (extractable, potentially degradable, undegradable, or slowly degradable) were unaffected by treatment. Lag time and kd varied among feeds. The kd was faster for all feeds (0.136/h) when incubated in dry-TMR1 cows compared with mid-TMR2 (0.097/h) or peak-TMR3 (0.098/h) cows, and no differences in lag time were detected. Calculated RUP, using estimated passage rates for each cow based on intake, differed between dry-TMR1 (0.382) and mid-TMR2 (0.559) or peak-TMR3 (0.626) cows, with a tendency for mid-TMR2 to be different from peak-TMR3. Using the average kd and lag time obtained from dry-TMR1 to calculate RUP for mid-TMR2 and peak-TMR3 cows using their passage rates reduced RUP values by 6.3 and 9.5 percentage units, respectively. Except for that of herring meal

  3. Human sperm degradation of zona pellucida proteins contributes to fertilization.

    Science.gov (United States)

    Saldívar-Hernández, Analilia; González-González, María E; Sánchez-Tusié, Ana; Maldonado-Rosas, Israel; López, Pablo; Treviño, Claudia L; Larrea, Fernando; Chirinos, Mayel

    2015-09-02

    The mammalian oocyte extracellular matrix known as the zona pellucida (ZP) acts as a barrier to accomplish sperm fusion with the female gamete. Although penetration of the ZP is a limiting event to achieve fertilization, this is one of the least comprehended stages of gamete interaction. Even though previous studies suggest that proteases of sperm origin contribute to facilitate the passage of sperm through the ZP, in human this process is not yet fully understood. The aim of this study was to determine the ability of human sperm to degrade recombinant human ZP (rhZPs) proteins and to characterize the proteases involved in this process. Purified rhZP2, rhZP3 and rhZP4 proteins were incubated with capacitated sperm and the proteolytic activity was determined by Western blot analysis. To further characterize the proteases involved, parallel incubations were performed in the presence of the protease inhibitors o-phenanthroline, benzamidine and MG-132 meant to block the activity of metalloproteases, serine proteases and the proteasome, respectively. Additionally, protease inhibitors effect on sperm-ZP binding was evaluated by hemizona assay. The results showed that rhZPs were hydrolyzed in the presence of capacitated sperm. O-phenanthroline inhibited the degradation of rhZP3, MG-132 inhibited the degradation of rhZP4 and benzamidine inhibited the degradation of the three proteins under investigation. Moreover, hemizona assays demonstrated that sperm proteasome inhibition impairs sperm interaction with human native ZP. This study suggests that sperm proteasomes could participate in the degradation of ZP, particularly of the ZP4 protein. Besides, metalloproteases may be involved in specific degradation of ZP3 while serine proteases may contribute to unspecific degradation of the ZP. These findings suggest that localized degradation of ZP proteins by sperm is probably involved in ZP penetration and may be of help in understanding the mechanisms of fertilization in humans.

  4. Rumen bacteria

    International Nuclear Information System (INIS)

    McSweeney, C.S.; Denman, S.E.; Mackie, R.I.

    2005-01-01

    The rumen is the most extensively studied gut community and is characterized by its high population density, wide diversity and complexity of interactions. This complex, mixed microbial culture is comprised of prokaryote organisms including methane-producing archaebacteria, eukaryote organisms, such as ciliate and flagellate protozoa, anaerobic phycomycete fungi and bacteriophage. Bacteria are predominant (up to 10 11 viable cells per g comprising 200 species) but a variety of ciliate protozoa occur widely (10 4 -10 6 /g distributed over 25 genera). The anaerobic fungi are also widely distributed (zoospore population densities of 10 2 -10 4 /g distributed over 5 genera). The occurrence of bacteriophage is well documented (10 7 -10 9 particles/g). This section focuses primarily on the widely used methods for the cultivation and the enumeration of rumen microbes, especially bacteria, which grow under anaerobic conditions. Methods that can be used to measure hydrolytic enzymes (cellulases, xylanases, amylases and proteinases) are also described, along with cell harvesting and fractionation procedures. Brief reference is also made to fungi and protozoa, but detailed explanations for culturing and enumerating these microbes is presented in Chapters 2.4 and 2.5

  5. Protein degradation: recognition of ubiquitinylated substrates

    DEFF Research Database (Denmark)

    Hartmann-Petersen, Rasmus; Gordon, Colin

    2004-01-01

    A cell-free system has been developed in budding yeast that provides direct evidence that the Dsk2/Dph1, Rad23/Rhp23 and Rpn10/Pus1 multi-ubiquitin-binding proteins, long implicated in substrate recognition and presentation to the 26S proteasome, actually fulfil such a role.......A cell-free system has been developed in budding yeast that provides direct evidence that the Dsk2/Dph1, Rad23/Rhp23 and Rpn10/Pus1 multi-ubiquitin-binding proteins, long implicated in substrate recognition and presentation to the 26S proteasome, actually fulfil such a role....

  6. Selectivity in protein degradation during sporulation of Bacillus subtilis

    International Nuclear Information System (INIS)

    Mitani, Takahiko; Kadota, Hajime

    1976-01-01

    The breakdown of cellular protein was investigated in Bacillus subtilis ATCC 6051 labeled with glycine-2- 3 H or L-phenylalanine-U- 14 C at the different stages of vegetative growth and sporulation. The growth of the culture was determined by measuring optical density at 660 nm. The heat-resistant spores were scored by plating after heating at 80 deg C for 10 minutes. A question whether the turnover of glycine-labeled protein is similar to that of phenylalanine-labeled protein was experimentally studied. The patterns obtained with the glycine-labeled protein were different from those of phenylalanine-labeled protein. This was not multiple turnover. The cellular protein which was labeled with glycine at an early stage of sporulation showed rapid degradation, but the degradation of the protein labeled with glycine at later stages did not occur at all. Another question whether the labeled glycine incorporated into cells at the different stages of growth and sporulation was present in the spore coat fraction of matured spores was studied. Experiment demonstrated that the glycine incorporated into cells at the late sporulation stage was mainly utilized for the biosynthesis of the spore coat protein. These data suggest that the spore coat protein which contains relatively large amount of glycine is rarely subject to further degradation. (Iwakiri, K.)

  7. Lignocellulose degradation, enzyme production and protein ...

    African Journals Online (AJOL)

    Microbial conversion of corn stover by white rot fungi has the potential to increase its ligninolysis and nutritional value, thereby transforming it into protein-enriched animal feed. Response surface methodology was applied to optimize conditions for the production of lignocellulolytic enzymes by Trametes versicolor during ...

  8. Prion protein degradation by lichens of the genus Cladonia

    Science.gov (United States)

    Bennett, James P.; Rodriguez, Cynthia M.; Johnson, Christopher J.

    2012-01-01

    It has recently been discovered that lichens contain a serine protease capable of degrading the pathogenic prion protein, the etiological agent of prion diseases such as sheep scrapie and cervid chronic wasting disease. Limited methods are available to degrade or inactivate prion disease agents, especially in the environment, and lichens or their serine protease could prove important for management of these diseases. Scant information is available regarding the presence or absence of the protease responsible for degrading prion protein (PrP) in lichen species and, in this study, we tested the hypothesis that PrP degradation activity in lichens is phylogenetically-based by testing 44 species of Cladonia lichens, a genus for which a significant portion of the phylogeny is well established. We categorized PrP degradation activity among the 44 species (high, moderate, low or none) and found that activity in Cladonia species did not correspond with phylogenetic position of the species. Degradation of PrP did correspond, however, with three classical taxonomic characters within the genus: species with brown apothecia, no usnic acid, and the presence of a cortex. Of the 44 species studied, 18 (41%) had either high or moderate PrP degradation activity, suggesting the protease may be frequent in this genus of lichens.

  9. Protein degradation during reconsolidation as a mechanism for memory reorganization

    Directory of Open Access Journals (Sweden)

    Bong-Kiun Kaang

    2011-02-01

    Full Text Available Memory is a reference formed from a past experience that is used to respond to present situations. However, the world is dynamic and situations change, so it is important to update the memory with new information each time it is reactivated in order to adjust the response in the future. Recent researches indicate that memory may undergo a dynamic process that could work as an updating mechanism. This process which is called reconsolidation involves destabilization of the memory after it is reactivated, followed by restabilization. Recently, it has been demonstrated that the initial destabilization process of reconsolidation requires protein degradation. Using protein degradation inhibition as a method to block reconsolidation, recent researches suggest that reconsolidation, especially the protein degradation-dependent destabilization process is necessary for memory reorganization.

  10. Proteolysis targeting peptide (PROTAP) strategy for protein ubiquitination and degradation.

    Science.gov (United States)

    Zheng, Jing; Tan, Chunyan; Xue, Pengcheng; Cao, Jiakun; Liu, Feng; Tan, Ying; Jiang, Yuyang

    2016-02-19

    Ubiquitination proteasome pathway (UPP) is the most important and selective way to degrade proteins in vivo. Here, a novel proteolysis targeting peptide (PROTAP) strategy, composed of a target protein binding peptide, a linker and a ubiquitin E3 ligase recognition peptide, was designed to recruit both target protein and E3 ligase and then induce polyubiquitination and degradation of the target protein through UPP. In our study, the PROTAP strategy was proved to be a general method with high specificity using Bcl-xL protein as model target in vitro and in cells, which indicates that the strategy has great potential for in vivo application. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Are animal models predictive for human postmortem muscle protein degradation?

    Science.gov (United States)

    Ehrenfellner, Bianca; Zissler, Angela; Steinbacher, Peter; Monticelli, Fabio C; Pittner, Stefan

    2017-11-01

    A most precise determination of the postmortem interval (PMI) is a crucial aspect in forensic casework. Although there are diverse approaches available to date, the high heterogeneity of cases together with the respective postmortal changes often limit the validity and sufficiency of many methods. Recently, a novel approach for time since death estimation by the analysis of postmortal changes of muscle proteins was proposed. It is however necessary to improve the reliability and accuracy, especially by analysis of possible influencing factors on protein degradation. This is ideally investigated on standardized animal models that, however, require legitimization by a comparison of human and animal tissue, and in this specific case of protein degradation profiles. Only if protein degradation events occur in comparable fashion within different species, respective findings can sufficiently be transferred from the animal model to application in humans. Therefor samples from two frequently used animal models (mouse and pig), as well as forensic cases with representative protein profiles of highly differing PMIs were analyzed. Despite physical and physiological differences between species, western blot analysis revealed similar patterns in most of the investigated proteins. Even most degradation events occurred in comparable fashion. In some other aspects, however, human and animal profiles depicted distinct differences. The results of this experimental series clearly indicate the huge importance of comparative studies, whenever animal models are considered. Although animal models could be shown to reflect the basic principles of protein degradation processes in humans, we also gained insight in the difficulties and limitations of the applicability of the developed methodology in different mammalian species regarding protein specificity and methodic functionality.

  12. Effects of rumen-undegradable protein on intake, performance, and mammary gland development in prepubertal and pubertal dairy heifers.

    Science.gov (United States)

    Silva, A L; Detmann, E; Dijkstra, J; Pedroso, A M; Silva, L H P; Machado, A F; Sousa, F C; Dos Santos, G B; Marcondes, M I

    2018-04-04

    The objective of this study was to evaluate the influence of different amounts of rumen-undegradable protein (RUP) on intake, N balance, performance, mammary gland development, carcass traits, and hormonal status of Holstein heifers at different physiological stages (PS). Sixteen prepubertal (PRE) heifers (initial BW = 106 ± 7.6 kg; age = 4.3 ± 0.46 mo) and 16 pubertal (PUB) heifers (initial BW = 224 ± 7.9 kg; age = 12.6 ± 0.45 mo) were used in an experiment over a period of 84 d. Four diets with increasing RUP contents (38, 44, 51, and 57% of dietary crude protein) and heifers at 2 PS (PRE or PUB) were used in a 4 × 2 factorial arrangement of treatments in a completely randomized design. Throughout the experiment, 2 digestibility trials were performed over 5 consecutive days (starting at d 36 and 78) involving feed and ort sampling and spot collections of feces and urine. At d 0 and 83, body ultrasound images were obtained for real-time carcass trait evaluation. The mammary gland was ultrasonically scanned at d 0 and every 3 wk during the experiment. Blood samples were taken at d 0 and 84 to determine serum concentrations of progesterone, estrogen, insulin-like growth factor I (IGF-I), and insulin. No interaction between PS and the level of RUP was found for any trait. Apparent digestibility of dry matter, organic matter, and neutral detergent fiber corrected for ash and protein was not affected by RUP level but was lower for PRE compared with PUB heifers. Sorting against neutral detergent fiber corrected for ash and protein (tendency only) and for crude protein was greater for PUB than PRE heifers. Pubertal heifers had greater average daily gain (905 vs. 505 g/d) and N retention (25.9 vs. 12.5 g/d) than PRE heifers. In addition, average daily gain and N retention were greatest at 51% RUP of dietary protein. Mammary ultrasonography indicated no effects of RUP amounts on mammary gland composition, whereas PRE heifers had greater pixel values than PUB

  13. Gene-centric metagenomics of the fiber-adherent bovine rumen microbiome reveals forage specific glycoside hydrolases.

    Science.gov (United States)

    Brulc, Jennifer M; Antonopoulos, Dionysios A; Miller, Margret E Berg; Wilson, Melissa K; Yannarell, Anthony C; Dinsdale, Elizabeth A; Edwards, Robert E; Frank, Edward D; Emerson, Joanne B; Wacklin, Pirjo; Coutinho, Pedro M; Henrissat, Bernard; Nelson, Karen E; White, Bryan A

    2009-02-10

    The complex microbiome of the rumen functions as an effective system for the conversion of plant cell wall biomass to microbial protein, short chain fatty acids, and gases. As such, it provides a unique genetic resource for plant cell wall degrading microbial enzymes that could be used in the production of biofuels. The rumen and gastrointestinal tract harbor a dense and complex microbiome. To gain a greater understanding of the ecology and metabolic potential of this microbiome, we used comparative metagenomics (phylotype analysis and SEED subsystems-based annotations) to examine randomly sampled pyrosequence data from 3 fiber-adherent microbiomes and 1 pooled liquid sample (a mixture of the liquid microbiome fractions from the same bovine rumens). Even though the 3 animals were fed the same diet, the community structure, predicted phylotype, and metabolic potentials in the rumen were markedly different with respect to nutrient utilization. A comparison of the glycoside hydrolase and cellulosome functional genes revealed that in the rumen microbiome, initial colonization of fiber appears to be by organisms possessing enzymes that attack the easily available side chains of complex plant polysaccharides and not the more recalcitrant main chains, especially cellulose. Furthermore, when compared with the termite hindgut microbiome, there are fundamental differences in the glycoside hydrolase content that appear to be diet driven for either the bovine rumen (forages and legumes) or the termite hindgut (wood).

  14. Feeding barley grain steeped in lactic acid modulates rumen fermentation patterns and increases milk fat content in dairy cows.

    Science.gov (United States)

    Iqbal, S; Zebeli, Q; Mazzolari, A; Bertoni, G; Dunn, S M; Yang, W Z; Ametaj, B N

    2009-12-01

    The objectives of the present in vivo and in situ trials were to evaluate whether feeding barley grain steeped in lactic acid (LA) would affect rumen fermentation patterns, in situ dry matter (DM) degradation kinetics, and milk production and composition in lactating dairy cows. The in vivo trial involved 8 rumen-fistulated Holstein cows fed once daily a total mixed ration containing rolled barley grain (27% in DM) steeped for 48 h in an equal quantity of tap water (CTR) or in 0.5% LA (TRT) in a 2 x 2 crossover design. The in situ trials consisted of incubation of untreated rolled barley grain in cows fed CTR or TRT diets and of incubation of 3 different substrates including CTR or barley grain steeped in 0.5% or 1.0% LA (TRT1 and TRT2, respectively) up to 72 h in the rumen. Results of the in vivo trial indicated that cows fed the TRT diet had greater rumen pH during most intensive fermentation phases at 10 and 12 h post-feeding. The latter effect was associated with a shorter duration in which rumen pH was below 5.8 for cows fed the TRT diet (2.4 h) compared with CTR diet (3.9 h). Furthermore, cows fed the TRT diet had lower concentrations of volatile fatty acids at 2 and 4 h post-feeding. In addition, concentrations of preprandial volatile fatty acids were lower in the rumen fluid of cows fed the TRT diet. Results also showed that molar proportion of acetate was lower, whereas propionate tended to increase by feeding cows the TRT diet. Cows fed the TRT diet demonstrated greater rumen in situ lag time of substrate DM degradation and a tendency to lower the fractional degradation rate. Other in situ results indicated a quadratic effect of LA on the effective rumen degradability of substrates whereby the latter variable was decreased from CTR to TRT1 but increased for TRT2 substrate. Although the diet did not affect actual milk yield, fat-corrected milk, percentages of milk protein, and lactose and concentration of milk urea nitrogen, cows fed the TRT diet increased

  15. Protein degradation and protein synthesis in long-term memory formation

    Directory of Open Access Journals (Sweden)

    Timothy J Jarome

    2014-06-01

    Full Text Available Long-term memory (LTM formation requires transient changes in the activity of intracellular signaling cascades that are thought to regulate new gene transcription and de novo protein synthesis in the brain. Consistent with this, protein synthesis inhibitors impair LTM for a variety of behavioral tasks when infused into the brain around the time of training or following memory retrieval, suggesting that protein synthesis is a critical step in LTM storage in the brain. However, evidence suggests that protein degradation mediated by the ubiquitin-proteasome system may also be a critical regulator of LTM formation and stability following retrieval. This requirement for increased protein degradation has been shown in the same brain regions in which protein synthesis is required for LTM storage. Additionally, increases in the phosphorylation of proteins involved in translational control parallel increases in protein polyubiquitination and the increased demand for protein degradation is regulated by intracellular signaling molecules thought to regulate protein synthesis during LTM formation. In some cases inhibiting proteasome activity can rescue memory impairments that result from pharmacological blockade of protein synthesis, suggesting that protein degradation may control the requirement for protein synthesis during the memory storage process. Results such as these suggest that protein degradation and synthesis are both critical for LTM formation and may interact to properly consolidate and store memories in the brain. Here, we review the evidence implicating protein synthesis and degradation in LTM storage and highlight the areas of overlap between these two opposing processes. We also discuss evidence suggesting these two processes may interact to properly form and store memories. LTM storage likely requires a coordinated regulation between protein degradation and synthesis at multiple sites in the mammalian brain.

  16. Metabolic and productive response to ruminal protein degradability in early lactation cows fed untreated or xylose-treated soybean meal-based diets.

    Science.gov (United States)

    Jahani-Moghadam, M; Amanlou, H; Nikkhah, A

    2009-12-01

    Effects of different dietary rumen undegradable (RUP) to degradable (RDP) protein ratios on ruminal nutrient degradation, feed intake, blood metabolites and milk production were determined in early lactation cows. Four multiparous (43 ± 5 days in milk) and four primiparous (40 ± 6 days in milk) tie-stall-housed Holstein cows were used in a duplicated 4 × 4 Latin square design with four 21-day periods. Each period had 14-day of adaptation and 7-day of sampling. Diets contained on a dry matter (DM) basis, 23.3% alfalfa hay, 20% corn silage and 56.7% concentrate. Cows were first offered alfalfa hay at 7:00, 15:00 and 23:00 hours, and 30 min after each alfalfa hay delivery were offered a mixture of corn silage and concentrate. Treatments were diets with RUP:RDP ratios of (i) 5.2:11.6 (control), (ii) 6.1:10.6, (iii) 7.1:9.5 and (iv) 8.1:8.5, on a dietary DM% basis. Different RUP:RDP ratios were obtained by partial and total replacement of untreated soybean meal (SBM) with xylose-treated SBM (XSBM). In situ study using three rumen-cannulated non-lactating cows showed that DM and crude protein (CP) of SBM had greater rapidly degradable fractions. The potentially degradable fractions were degraded more slowly in XSBM. Treatment cows produced greater milk, protein, lactose, solids-non-fat and total solids than control cows. Increasing RUP:RDP reduced blood urea linearly. Feed costs dropped at RUP:RDP ratios of 6.1:10.6 and 7.1:9.5, but not at 8.1:8.5, compared with the 5.2:11.6 ratio. Intake of DM and CP, rumen pH, blood glucose, albumin and total protein, faecal and urine pH, changes in body weight and body condition score, and milk lactose and solids-non-fat percentages did not differ among treatments. Results provide evidence that increasing dietary RUP:RDP ratio from 5.2:11.6 to 7.1:9.5 optimizes nitrogen metabolism and milk production and reduces feed costs in early lactation cows. Reduced blood urea suggests reprodutive benefits.

  17. Degradation of Akt using protein-catalyzed capture agents.

    Science.gov (United States)

    Henning, Ryan K; Varghese, Joseph O; Das, Samir; Nag, Arundhati; Tang, Grace; Tang, Kevin; Sutherland, Alexander M; Heath, James R

    2016-04-01

    Abnormal signaling of the protein kinase Akt has been shown to contribute to human diseases such as diabetes and cancer, but Akt has proven to be a challenging target for drugging. Using iterative in situ click chemistry, we recently developed multiple protein-catalyzed capture (PCC) agents that allosterically modulate Akt enzymatic activity in a protein-based assay. Here, we utilize similar PCCs to exploit endogenous protein degradation pathways. We use the modularity of the anti-Akt PCCs to prepare proteolysis targeting chimeric molecules that are shown to promote the rapid degradation of Akt in live cancer cells. These novel proteolysis targeting chimeric molecules demonstrate that the epitope targeting selectivity of PCCs can be coupled with non-traditional drugging moieties to inhibit challenging targets. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.

  18. Evaluation of the effect of fat content of sunflower meal on rumen ...

    African Journals Online (AJOL)

    Yomi

    2012-01-03

    Jan 3, 2012 ... addition of yellow grease and corn oil increased in vitro rumen degradation and gas production. ... Culturing of rumen anaerobic fungi. Rumen anaerobic fungi were isolated from wheat straw residues, ... concentrate : forage diet (corn grain, barley grain and wheat bran : sugarcane silage, corn silage, alfalfa ...

  19. Rumen degradation of oil palm fronds is improved through pre-digestion with white rot fungi but not through supplementation with yeast or enzymes

    NARCIS (Netherlands)

    Hassim, H.A.; Lourenco, M.; Goh, Y.M.; Baars, J.J.P.; Fievez, V.

    2012-01-01

    Rumen fermentation kinetics of oil palm fronds (OPF) supplemented or not with enzymes (Hemicell® or Allzyme SSF®) or yeasts (Levucell®SC or Yea-Sacc®) were studied through an in vitro gas production test (96 h) (exp. 1). In exp. 2, enzymes were supplemented to OPF pre-treated during 3 or 9 wk with

  20. Metagenomic insights into lignocellulose-degrading genes through Illumina-based de novo sequencing of the microbiome in Vietnamese native goats' rumen

    NARCIS (Netherlands)

    Do, Thi Huyen; Le, Ngoc Giang; Dao, Trong Khoa; Nguyen, Thi Mai Phuong; Le, Tung Lam; Luu, Han Ly; Nguyen, Khanh Hoang Viet; Nguyen, Van Lam; Le, Lan Anh; Phung, Thu Nguyet; van Straalen, Nico M; Roelofs, Dick; Truong, Nam Hai

    2018-01-01

    The scarcity of enzymes having an optimal activity in lignocellulose deconstruction is an obstacle for industrial-scale conversion of cellulosic biomass into biofuels. With the aim of mining novel lignocellulolytic enzymes, a ~9 Gb metagenome of bacteria in Vietnamese native goats' rumen was

  1. Impact of protein uptake and degradation on recombinant protein secretion in yeast

    DEFF Research Database (Denmark)

    Tyo, Keith E. J.; Liu, Zihe; Magnusson, Ylva

    2014-01-01

    Protein titers, a key bioprocessing metric, depend both on the synthesis of protein and the degradation of protein. Secreted recombinant protein production in Saccharomyces cerevisiae is an attractive platform as minimal media can be used for cultivation, thus reducing fermentation costs...... and transcriptomics, we identify metabolic and regulatory markers that are consistent with uptake of whole proteins by endocytosis, followed by intracellular degradation and catabolism of substituent amino acids. Uptake and degradation of recombinant protein products may be common in S. cerevisiae protein secretion...... and simplifying downstream purification, compared to other systems that require complex media. As such, engineering S. cerevisiae to improve titers has been then the subject of significant attention, but the majority of previous efforts have been focused on improving protein synthesis. Here, we characterize...

  2. Effect of Carbohydrate Source and Cottonseed Meal Level in the Concentrate on Feed Intake, Nutrient Digestibility, Rumen Fermentation and Microbial Protein Synthesis in Swamp Buffaloes

    Directory of Open Access Journals (Sweden)

    M. Wanapat

    2013-07-01

    Full Text Available The objective of this study was to investigate the effect of carbohydrate source and cottonseed meal level in the concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in swamp buffaloes. Four, 4-yr old rumen fistulated swamp buffaloes were randomly assigned to receive four dietary treatments according to a 2×2 factorial arrangement in a 4×4 Latin square design. Factor A was carbohydrate source; cassava chip (CC and CC+rice bran at a ratio 3:1 (CR3:1, and factor B was level of cottonseed meal (CM; 109 g CP/kg (LCM and 328 g CP/kg (HCM in isonitrogenous diets (490 g CP/kg. Buffaloes received urea-treated rice straw ad libitum and supplemented with 5 g concentrate/kg BW. It was found that carbohydrate source did not affect feed intake, nutrient intake, digested nutrients, nutrient digestibility, ammonia nitrogen concentration, fungi and bacterial populations, or microbial protein synthesis (p>0.05. Ruminal pH at 6 h after feeding and the population of protozoa at 4 h after feeding were higher when buffalo were fed with CC than in the CR3:1 treatment (p0.05. Based on this experiment, concentrate with a low level of cottonseed meal could be fed with cassava chips as an energy source in swamp buffalo receiving rice straw.

  3. In situ ruminal degradability of protein feeds with distinct physical ...

    African Journals Online (AJOL)

    Journal Home > Vol 47, No 1 (2017) > ... of protein feeds with different physical forms (meal versus grain) through a meta-analysis study. ... grains with the meal forms of soybean, peanut, sunflower, cottonseed, and corn gluten. The degradation parameters of dry matter did not differ significantly between meals and grains.

  4. Rate and extent of ruminal degradation of crude protein from ...

    African Journals Online (AJOL)

    Predicted crude protein degradation was calculated at rate constants for outflow of 0.04 and 0.06/h respect- ively. ... as buffers, an ionophore and an antibiotic according to general .... the non-bird resistant ('sweet') varieties. Ruminal .... have been affected by both the particle type and the math- ematical model we used.

  5. Endocytosis and intracellular protein degradation in cystic fibrosis fibroblasts

    International Nuclear Information System (INIS)

    Jessup, W.; Dean, R.T.

    1983-01-01

    Normal rates of pinocytosis of [ 3 H]sucrose were measured in cystic fibrosis fibroblasts, and were not affected by the addition of cystic fibrosis serum. Bulk protein degradation (a significant proportion of which occurs intralysosomally following autophagy) and its regulation by growth state were apparently identical in normal and cystic fibrosis cultures. (Auth.)

  6. Proteasomal and lysosomal protein degradation and heart disease.

    Science.gov (United States)

    Wang, Xuejun; Robbins, Jeffrey

    2014-06-01

    In the cell, the proteasome and lysosomes represent the most important proteolytic machineries, responsible for the protein degradation in the ubiquitin-proteasome system (UPS) and autophagy, respectively. Both the UPS and autophagy are essential to protein quality and quantity control. Alterations in cardiac proteasomal and lysosomal degradation are remarkably associated with most heart disease in humans and are implicated in the pathogenesis of congestive heart failure. Studies carried out in animal models and in cell culture have begun to establish both sufficiency and, in some cases, the necessity of proteasomal functional insufficiency or lysosomal insufficiency as a major pathogenic factor in the heart. This review article highlights some recent advances in the research into proteasome and lysosome protein degradation in relation to cardiac pathology and examines the emerging evidence for enhancing degradative capacities of the proteasome and/or lysosome as a new therapeutic strategy for heart disease. This article is part of a Special Issue entitled "Protein Quality Control, the Ubiquitin Proteasome System, and Autophagy". Copyright © 2013 Elsevier Ltd. All rights reserved.

  7. Establishment of a ruminal protein degradation data base for dairy ...

    African Journals Online (AJOL)

    Establishment of a ruminal protein degradation data base for dairy cattle using the in situ polyester bag technique. 2. Energy sources. LJ Erasmus, J Prinsloo, PM Botha, HH Meissner. Abstract. No Abstract. Full Text: EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT.

  8. Establishment of a ruminal protein degradation data base for dairy ...

    African Journals Online (AJOL)

    Establishment of a ruminal protein degradation data base for dairy cattle using the in situ polyester bag technique. 3. Roughages. LJ Erasmus, J Prinsloo, PM Botha, HH Meissner. Abstract. No Abstract. Full Text: EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT.

  9. Rumen microbial community composition varies with diet and host, but a core microbiome is found across a wide geographical range

    NARCIS (Netherlands)

    Henderson, G.; Cox, F.; Ganesh, S.; Jonker, A.; Young, W.; Janssen, P.H.; Bannink, A.; Dieho, K.; Dijkstra, J.

    2015-01-01

    Ruminant livestock are important sources of human food and global greenhouse gas emissions. Feed degradation and methane formation by ruminants rely on metabolic interactions between rumen microbes and affect ruminant productivity. Rumen and camelid foregut microbial community composition was

  10. Light-induced protein degradation in human-derived cells.

    Science.gov (United States)

    Sun, Wansheng; Zhang, Wenyao; Zhang, Chao; Mao, Miaowei; Zhao, Yuzheng; Chen, Xianjun; Yang, Yi

    2017-05-27

    Controlling protein degradation can be a valuable tool for posttranslational regulation of protein abundance to study complex biological systems. In the present study, we designed a light-switchable degron consisting of a light oxygen voltage (LOV) domain of Avena sativa phototropin 1 (AsLOV2) and a C-terminal degron. Our results showed that the light-switchable degron could be used for rapid and specific induction of protein degradation in HEK293 cells by light in a proteasome-dependent manner. Further studies showed that the light-switchable degron could also be utilized to mediate the degradation of secreted Gaussia princeps luciferase (GLuc), demonstrating the adaptability of the light-switchable degron in different types of protein. We suggest that the light-switchable degron offers a robust tool to control protein levels and may serves as a new and significant method for gene- and cell-based therapies. Copyright © 2017 Elsevier Inc. All rights reserved.

  11. Cinética ruminal da degradação de nutrientes da silagem de milho em ambiente ruminal inoculado com diferentes aditivos Ruminal degradation kinetics of corn silage in bulls inoculated with different additives in the rumen

    Directory of Open Access Journals (Sweden)

    Pedro Andrade Katsuki

    2006-12-01

    Full Text Available Objetivou-se avaliar a cinética ruminal da degradação de MS, PB e FDN da silagem de milho em ambiente ruminal inoculado com diferentes aditivos. Utilizou-se um delineamento em quadrado latino 4 x 4, com quatro bovinos holandeses e quatro períodos de incubação, em ambiente ruminal adaptado ou não com diferentes aditivos alimentares. Foram testados os seguintes tratamentos: SCL - silagem de milho em ambiente ruminal sem inoculação de aditivo; SBL - silagem de milho em ambiente ruminal inoculado com 5 g de produto comercial contendo bactérias ruminais e intestinais liofilizadas (Ruminobacter amylophilum: 3,0 x 10(11 ufc/kg; Fibrobacter succinogenes: 3,0 x 10(11 ufc/kg; Succinovibrio dextrinsolvens: 4,4 x 10(11 ufc/kg; Bacillus cereus: 3,5 x 10(11 ufc/kg; Lactobacillus acidophilus: 3,5 x 10(11 ufc/kg e Streptococcus faecium: 3,5 x 10(11 ufc/kg; SEC - silagem de milho em ambiente ruminal inoculado com 15 g de produto comercial contendo enzimas celulolíticas (xilanase 10%; e SMS - silagem de milho em ambiente ruminal inoculado com 3 g de produto comercial contendo monensina sódica. Os tratamentos SBL e SEC não afetaram a fração potencialmente degradável (b dos nutrientes avaliados da silagem de milho. A monensina sódica reduziu a fração (b da MS (51,01% e a degradabilidade potencial da silagem de milho (72,33%. Entre os aditivos estudados, a monensina sódica proporcionou a maior fração não-degradável da FDN (45,57%, reduzindo o desaparecimento desta fração a partir de 48 horas de incubação intra-ruminal. Os diferentes aditivos, nas concentrações estudadas, não proporcionaram melhora na degradabilidade efetiva da MS, PB e FDN da silagem de milho.Four bulls fitted with ruminal cannula were used in a 4 x 4 Latin square design to evaluate the effects of different ruminally inoculated additives on the degradation kinetics of DM, CP, and NDF of corn silage (CS. The treatments were: control CS incubated in rumen with no

  12. Metagenomic Characterization and Biochemical Analysis of Cellulose-Degrading Bacterial Communities from Sheep Rumen, Termite Hindgut, Decaying Plant Materials, and Soil

    Science.gov (United States)

    2016-01-04

    ethanol production (Mielenz 2011). The combustion of bioethanol results in clean emission of heat , steam, and most importantly carbon dioxide. Next...allows plants to stand   3   upright and grow toward the sun, withstand environmental stresses , and to defend against herbivores. Infamously known...However, a metagenomic study in cow has failed to identify individual microorganismal groups in the rumen (Hess et al. 2011) Decaying plant and Soil

  13. Short communication: Using diurnal patterns of (13)C enrichment of CO2 to evaluate the effects of nitrate and docosahexaenoic acid on fiber degradation in the rumen of lactating dairy cows.

    Science.gov (United States)

    Klop, G; Bannink, A; Dieho, K; Gerrits, W J J; Dijkstra, J

    2016-09-01

    Nitrate decreases enteric CH4 production in ruminants, but may also negatively affect fiber degradation. In this experiment, 28 lactating Holstein dairy cows were grouped into 7 blocks. Within blocks, cows were randomly assigned to 1 of 4 isonitrogenous treatments in a 2×2 factorial arrangement: control (CON); NO3 [21g of nitrate/kg of dry matter (DM)]; DHA [3g of docosahexaenoic acid (DHA)/kg of DM]; or NO3+DHA (21g of nitrate/kg of DM and 3g of DHA/kg of DM). Cows were fed a total mixed ration consisting of 21% grass silage, 49% corn silage, and 30% concentrates on a DM basis. Based on the difference in natural (13)C enrichment and neutral detergent fiber and starch content between grass silage and corn silage, we investigated whether a negative effect on rumen fiber degradation could be detected by evaluating diurnal patterns of (13)C enrichment of exhaled carbon dioxide. A significant nitrate × DHA interaction was found for neutral detergent fiber digestibility, which was reduced on the NO3 treatment to an average of 55%, as compared with 61, 64, and 65% on treatments CON, DHA, and NO3+DHA, respectively. Feeding nitrate, but not DHA, resulted in a pronounced increase in (13)C enrichment of CO2 in the first 3 to 4 h after feeding only. Results support the hypothesis that effects of a feed additive on the rate of fiber degradation in the rumen can be detected by evaluating diurnal patterns of (13)C enrichment of CO2. To be able to detect this, the main ration components have to differ considerably in fiber and nonfiber carbohydrate content as well as in natural (13)C enrichment. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  14. Comparison of Rumen Degradation Characteristics between Pennisetum sp.and Commonly Used Roughages for Dairy Cows%禾王草与奶牛常用粗饲料瘤胃降解特性的对比研究

    Institute of Scientific and Technical Information of China (English)

    马健; 刘艳芳; 杜云; 李胜利; 余雄

    2016-01-01

    ( DM) , crude protein ( CP) , neutral detergent fiber ( NDF) and acid detergent fiber ( ADF) . The results showed as follows:1) the content of CP in Pennisetum sp.hay was significantly high than that in dried Chinese wildye ( P0.05) , but was sig-nificantly higher than that of 2.5 m Pennisetum sp.silage, 2.0 m Pennisetum sp.hay, whole corn silage, 2.5 m Pennisetum sp.hay and dried Chinese wildye ( P0.05), and was significantly higher than that of other roughages (P<0.05); ADF effective degradability of Pennisetum sp.silage at 72 h was significantly higher than that of other roughers (P<0.05).It is concluded that Pennisetum sp.has a potential as a new non-conventional feed resource for ruminants, silage can save the nutrition composition, and can improve the rumen degradation of nutrition composition.

  15. Effect of Carbohydrate Sources and Levels of Cotton Seed Meal in Concentrate on Feed Intake, Nutrient Digestibility, Rumen Fermentation and Microbial Protein Synthesis in Young Dairy Bulls

    Directory of Open Access Journals (Sweden)

    M. Wanapat

    2013-04-01

    Full Text Available The objective of this study was to investigate the effect of levels of cottonseed meal with various carbohydrate sources in concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in dairy bulls. Four, 6 months old dairy bulls were randomly assigned to receive four dietary treatments according to a 2×2 factorial arrangement in a 4×4 Latin square design. Factor A was carbohydrate source; cassava chip (CC and cassava chip+rice bran in the ratio of 3:1 (CR3:1, and factor B was cotton seed meal levels in the concentrate; 109 g CP/kg (LCM and 328 g CP/kg (HCM at similar overall CP levels (490 g CP/kg. Bulls received urea-lime treated rice straw ad libitum and were supplemented with 10 g of concentrate/kg BW. It was found that carbohydrate source and level of cotton seed meal did not have significant effects on ruminal pH, ammonia nitrogen concentration, microbial protein synthesis or feed intake. Animals which received CC showed significantly higher BUN concentration, ruminal propionic acid and butyric acid proportions, while dry matter, organic matter digestibility, populations of total viable bacteria and proteolytic bacteria were lower than those in the CR3:1 treatment. The concentration of total volatile fatty acids was higher in HCM than LCM treatments, while the concentration of butyric acid was higher in LCM than HCM treatments. The population of proteolytic bacteria with the LCM treatments was higher than the HCM treatments; however other bacteria groups were similar among the different levels of cotton seed meal. Bulls which received LCM had higher protein digestibility than those receiving HCM. Therefore, using high levels of cassava chip and cotton seed meal might positively impact on energy and nitrogen balance for the microbial population in the rumen of the young dairy bull.

  16. Exploitation of dietary tannins to improve rumen metabolism and ruminant nutrition.

    Science.gov (United States)

    Patra, Amlan K; Saxena, Jyotisna

    2011-01-15

    Tannins (hydrolysable and condensed tannin) are polyphenolic polymers of relatively high molecular weight with the capacity to form complexes mainly with proteins due to the presence of a large number of phenolic hydroxyl groups. They are widely distributed in nutritionally important forage trees, shrubs and legumes, cereals and grains, which are considered as anti-nutritional compounds due to their adverse effects on intake and animal performance. However, tannins have been recognised to modulate rumen fermentation favourably such as reducing protein degradation in the rumen, prevention of bloat, inhibition of methanogenesis and increasing conjugated linoleic acid concentrations in ruminant-derived foods. The inclusion of tannins in diets has been shown to improve body weight and wool growth, milk yields and reproductive performance. However, the beneficial effects on rumen modulation and animal performance have not been consistently observed. This review discusses the effects of tannins on nitrogen metabolism in the rumen and intestine, and microbial populations (bacteria, protozoa, fungi and archaea), metabolism of tannins, microbial tolerance mechanisms to tannins, inhibition of methanogenesis, ruminal biohydrogenation processes and performance of animals. The discrepancies of responses of tannins among different studies are attributed to the different chemical structures (degree of polymerisation, procyanidins to propdelphinidins, stereochemistry and C-C bonding) and concentrations of tannins, and type of diets. An establishment of structure-activity relationship would be required to explain differences among studies and obtain consistent beneficial tannin effects. Copyright © 2010 Society of Chemical Industry.

  17. Protein aggregation and degradation during iodine labeling and its consequences for protein adsorption to biomaterials

    DEFF Research Database (Denmark)

    Holmberg, Maria; Jensen, Karin Bagger Stibius; Ndoni, Sokol

    2007-01-01

    Protein adsorption on modified and unmodified polymer surfaces investigated through radiolabeling experiments showed a tendency for higher than expected albumin and immunoglobulin G (IgG) adsorption. Possible enhanced protein aggregation and degradation caused by the iodine labeling method used w...

  18. Direct ubiquitin independent recognition and degradation of a folded protein by the eukaryotic proteasomes-origin of intrinsic degradation signals.

    Directory of Open Access Journals (Sweden)

    Amit Kumar Singh Gautam

    Full Text Available Eukaryotic 26S proteasomes are structurally organized to recognize, unfold and degrade globular proteins. However, all existing model substrates of the 26S proteasome in addition to ubiquitin or adaptor proteins require unstructured regions in the form of fusion tags for efficient degradation. We report for the first time that purified 26S proteasome can directly recognize and degrade apomyoglobin, a globular protein, in the absence of ubiquitin, extrinsic degradation tags or adaptor proteins. Despite a high affinity interaction, absence of a ligand and presence of only helices/loops that follow the degradation signal, apomyoglobin is degraded slowly by the proteasome. A short floppy F-helix exposed upon ligand removal and in conformational equilibrium with a disordered structure is mandatory for recognition and initiation of degradation. Holomyoglobin, in which the helix is buried, is neither recognized nor degraded. Exposure of the floppy F-helix seems to sensitize the proteasome and primes the substrate for degradation. Using peptide panning and competition experiments we speculate that initial encounters through the floppy helix and additional strong interactions with N-terminal helices anchors apomyoglobin to the proteasome. Stabilizing helical structure in the floppy F-helix slows down degradation. Destabilization of adjacent helices accelerates degradation. Unfolding seems to follow the mechanism of helix unraveling rather than global unfolding. Our findings while confirming the requirement for unstructured regions in degradation offers the following new insights: a origin and identification of an intrinsic degradation signal in the substrate, b identification of sequences in the native substrate that are likely to be responsible for direct interactions with the proteasome, and c identification of critical rate limiting steps like exposure of the intrinsic degron and destabilization of an unfolding intermediate that are presumably

  19. Model for Stress-induced Protein Degradation in Lemna minor1

    Science.gov (United States)

    Cooke, Robert J.; Roberts, Keith; Davies, David D.

    1980-01-01

    Transfer of Lemna minor fronds to adverse or stress conditions produces a large increase in the rate of protein degradation. Cycloheximide partially inhibits stress-induced protein degradation and also partially inhibits the protein degradation which occurs in the absence of stress. The increased protein degradation does not appear to be due to an increase in activity of soluble proteolytic enzymes. Biochemical evidence indicates that stress, perhaps acting via hormones, affects the permeability of certain membranes, particularly the tonoplast. A general model for stress-induced protein degradation is presented in which changes in membrane properties allow vacuolar proteolytic enzymes increased access to cytoplasmic proteins. PMID:16661588

  20. Nitrogen metabolism in the rumen and its measurement

    International Nuclear Information System (INIS)

    Nolan, J.V.; Leng, R.A.

    1983-01-01

    Methods are needed to maximize the synthesis of microbial protein in the rumen from readily available, inexpensive (usually non-protein N) sources and thereby to reduce the requirements for true protein in the diet. Some currently available in vitro and in vivo methods for estimating microbial protein synthesis in the rumen are discussed. The factors that alter maintenance ATP requirements of microorganisms, and thereby potentially alter the efficiency of cell growth per unit of fermented organic matter (Ysub(ATP)), are discussed: e.g. continuity and level of supply of substrates; dietary and recycled N and other nutrients; dilution rate; the presence of large protozoal populations in the rumen; cell lysis and N cycling in the rumen. Quantitative studies of these factors have been made by using a variety of isotope tracer techniques and have been applied to a quantitative model of N transactions in the rumen. (author)

  1. Formation of methylamine by rumen microorganisms

    International Nuclear Information System (INIS)

    Itabashi, Hisao; Kandatsu, Makoto.

    1978-01-01

    An unknown ninhydrin positive peak on the chromatograms of amino acid analyzer of alkalified rumen fluid distillate of goats was isolated as DNP-derivative and identified as methylamine. Under normal feeding condition, its concentration in the rumen ranged 0.1-3.9 mgN/100 ml of rumen fluid and the proportion of methylamine in total volatile base, or apparent ammonia, ranged 0.5-13% during post-feeding. When ammonium salt was administered into the rumen with hay-concentrate ration, these values were increased up to 8.1 mgN/100 ml and 25.8% respectively. Concentrations of ammonia and methylamine when aspartic acid or alanine was administered into the rumen in place of concentrate mixture (control) were not markedly different from the control. In the case of arginine, glutamic acid or glycine administration, these concentrations were depressed as compared to the control. There were no distinct differences in the concentration of methylamine between the faunated and unfaunated goats. 14 C from 14 C-chlorella protein hydrolyzates, U- 14 C-alanine, 2- 14 C-glycine or 14 C-sodium bicarbonate was incorporated into methylamine in invitro incubation with rumen micro-organisms. When the washed suspensions of rumen bacteria or protozoa were incubated with 14 C-chlorella protein hydrolyzates, the radioactivity in methylamine appeared only in the case of bacteria suspensions. After the addition of 15 N-ammonium citrate into the rumen, the incorporation of 15 N into methylamine was observed during 1-9 hr. (auth.)

  2. Common Prairie feeds with different soluble and insoluble fractions used for CPM diet formulation in dairy cattle: impact of carbohydrate-protein matrix structure on protein and other primary nutrient digestion.

    Science.gov (United States)

    Peng, Quanhui; Wang, Zhisheng; Zhang, Xuewei; Yu, Peiqiang

    2014-01-01

    An experiment was conducted to investigate the relationship of carbohydrates molecular spectral characteristics to rumen degradability of primary nutrients in Prairie feeds in dairy cattle. In total, 12 different types of feeds were selected, each type of feed was from three different source with total 37 samples. Six types of them were energy-sourced feeds and the others were protein-sourced feeds. The carbohydrates molecular spectral intensity of various functional groups were collected using Fourier transform infrared attenuated total reflectance (ATR-FT/IR) spectroscopy. In the in situ study, the results showed that the rumen digestibility and digestible fractions of primary nutrients (DM, OM, NCP, and CP) were significantly different (P<0.05) among the feeds. The spectral bands features were significantly different (P<0.05) among the feeds. Spectral intensities of A_Cell, H_1415 and H_1370 were weakly positively correlated with in situ rumen digestibility and digestible fractions of DM, OM and NCP. Spectral intensities of H_1150, H_1015, A_1, and A_3 were weakly negatively associated with in situ rumen degradation of CP. Spectral intensities of A_1240 and H_1240, mainly associated with cellulosic compounds, were correlated with rumen CP degradation. The multiple regression analysis demonstrated that the spectral intensities of A_3 and H_1415 played the most important role and could be used as a potential tool to predict rumen protein degradation of feeds in dairy cattle. In conclusion, this study showed that the carbohydrates as a whole have an effect on protein rumen degradation, rather than cellulose alone, indicating carbohydrate-protein matrix structure impact protein utilization in dairy cattle. The non-invasive molecular spectral technique (ATR-FT/IR) could be used as a rapid potential tool to predict rumen protein degradation of feedstuffs by using molecular spectral bands intensities in carbohydrate fingerprint region. Copyright © 2013 Elsevier B

  3. Comparative metabolite fingerprinting of the rumen system during colonisation of three forage grass (Lolium perenne L. varieties.

    Directory of Open Access Journals (Sweden)

    Alison H Kingston-Smith

    Full Text Available The rumen microbiota enable ruminants to degrade complex ligno-cellulosic compounds to produce high quality protein for human consumption. However, enteric fermentation by domestic ruminants generates negative by-products: greenhouse gases (methane and environmental nitrogen pollution. The current lack of cultured isolates representative of the totality of rumen microbial species creates an information gap about the in vivo function of the rumen microbiota and limits our ability to apply predictive biology for improvement of feed for ruminants. In this work we took a whole ecosystem approach to understanding how the metabolism of the microbial population responds to introduction of its substrate. Fourier Transform Infra Red (FTIR spectroscopy-based metabolite fingerprinting was used to discriminate differences in the plant-microbial interactome of the rumen when using three forage grass varieties (Lolium perenne L. cv AberDart, AberMagic and Premium as substrates for microbial colonisation and fermentation. Specific examination of spectral regions associated with fatty acids, amides, sugars and alkanes indicated that although the three forages were apparently similar by traditional nutritional analysis, patterns of metabolite flux within the plant-microbial interactome were distinct and plant genotype dependent. Thus, the utilisation pattern of forage nutrients by the rumen microbiota can be influenced by subtleties determined by forage genotypes. These data suggest that our interactomic approach represents an important means to improve forages and ultimately the livestock environment.

  4. Rumen bypass nutrients: Manipulation and implications

    International Nuclear Information System (INIS)

    Leng, R.A.; Nolan, J.V.; Preston, T.R.

    1983-01-01

    The feeds available for ruminants in developing countries are either agro-industrial by-products or specially grown forage crops. Many of these feeds are low in protein and require supplementation with non-protein N (NPN) to maintain efficient rumen function and digestibility. The principles for utilizing high energy, low protein feeds by ruminants are discussed in relation to the supply of NPN, the establishment of efficient rumen function, maximizing feed intake by means of supplements, and increasing total energy and protein intake by using supplements which bypass the rumen. To illustrate it the application of these principles to feeding systems based on molasses, chopped whole sugar cane and derinded sugar cane is discussed. The implications of the principles in increasing the feeding value of straw are also discussed. (author)

  5. Incorporation of 15N and 14C into amino acids of bacterial and protozoal protein in the rumen of the cow on urea-rich feed

    Directory of Open Access Journals (Sweden)

    Eeva-Liisa Syväoja

    1979-01-01

    Full Text Available The utilization of the non-protein nitrogen and carbon of feed by rumen microorganisms for the synthesis of protein was studied by administering [U-14C] sucrose and 15NH4Cl to a cow on urea-rich, low-protein feed. By studying the labelling of the protozoa and bacteria and the amino acids isolated from them at intervals up to 48 hours afterwards, it was found that the bacteria synthesized amino acids from nonprotein nitrogen much more rapidly and effectively than the protozoa. Also the labelling of the carbon in the amino acids of the bacteria was more rapid than in the protozoa. In both protozoa and bacteria there was intracellular storage of [14C] sucrose. Of the bacterial amino acids the most vigorous 14C labelling was found in Glu, Arg, Lys, Val and Ala and the weakest labelling in Gly, His and Ser. Of the protozoal amino acids Ala, Asp, Glu, Leu and Lys had the highest labelling and Pro, Gly, His and Phe the lowest. In the bacterial protein the labelling of Pro and Arg was ten times that of the corresponding protozoal amino acids, and Asp, Ser and Ala four times. After the 15NH4Cl dose the half-life of 15N in the rumen fluid was estimated to be 3.3 h. Labelled ammonium nitrogen was about 11 —15 % of the bacterial nitrogen and 2—3 % of the protozoal nitrogen after 1 h. Of the protozoal amino acids Ala, Glu, Val, Asp and Met had the most vigorous labelling, and of the bacterial amino acids Glu, Asp, Ser, He and Tyr. The slowest incorporation of ammonium nitrogen was into His, Pro, Arg and Gly in both bacteria and protozoa. The labelling of the bacterial amino acids was approximately 7—8 times more vigorous than that of the protozoal amino acids. The labelling of Ala was only 4 times, and that of Val, Met and Glu 5 times more vigorous than with protozoal protein. The pathway of histidine synthesis seemed to be restricted in both bacteria and protozoa and therefore may be a limiting factor in protein synthesis, particularly in cows fed

  6. The relationship between protein synthesis and protein degradation in object recognition memory.

    Science.gov (United States)

    Furini, Cristiane R G; Myskiw, Jociane de C; Schmidt, Bianca E; Zinn, Carolina G; Peixoto, Patricia B; Pereira, Luiza D; Izquierdo, Ivan

    2015-11-01

    For decades there has been a consensus that de novo protein synthesis is necessary for long-term memory. A second round of protein synthesis has been described for both extinction and reconsolidation following an unreinforced test session. Recently, it was shown that consolidation and reconsolidation depend not only on protein synthesis but also on protein degradation by the ubiquitin-proteasome system (UPS), a major mechanism responsible for protein turnover. However, the involvement of UPS on consolidation and reconsolidation of object recognition memory remains unknown. Here we investigate in the CA1 region of the dorsal hippocampus the involvement of UPS-mediated protein degradation in consolidation and reconsolidation of object recognition memory. Animals with infusion cannulae stereotaxically implanted in the CA1 region of the dorsal hippocampus, were exposed to an object recognition task. The UPS inhibitor β-Lactacystin did not affect the consolidation and the reconsolidation of object recognition memory at doses known to affect other forms of memory (inhibitory avoidance, spatial learning in a water maze) while the protein synthesis inhibitor anisomycin impaired the consolidation and the reconsolidation of the object recognition memory. However, β-Lactacystin was able to reverse the impairment caused by anisomycin on the reconsolidation process in the CA1 region of the hippocampus. Therefore, it is possible to postulate a direct link between protein degradation and protein synthesis during the reconsolidation of the object recognition memory. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Minerals and rumen function

    International Nuclear Information System (INIS)

    Smith, R.H.

    1984-01-01

    The mechanisms are discussed of some clinical disorders, characteristic only of ruminants and related to the effects of abnormal mineral intake on rumen function. With particular attention to tropical conditions, consideration is given to: (a) the possible effects of phosphorus deficiency on rumen microbial activity; (b) the depression of rumen microbial synthesis in sulphur deficiency; (c) the inhibition of magnesium absorption from the forestomachs; and (d) the involvement of the rumen microorganisms in leading to copper and vitamin B 12 deficiencies as a result of low intakes of cobalt. (author)

  8. In vitro evaluation of salinomycin addition in wheat straw based total mixed diets on rumen fermentation, methanogenesis and dry matter degradability in buffalo

    Directory of Open Access Journals (Sweden)

    Sunil K. Sirohi

    Full Text Available Aim: The aim of the current study was to evaluate the effect of salinomycin in vitro on methanogenesis and rumen fermentation. Materials and Methods: Different levels of (0,10, 15 and 20 ppm salinomycin were checked for their effect on in vitro methanogenesis and rumen fermentation on three wheat straw based diets i.e. low fiber diet (LFD, 40R:60C, medium fiber diet (MFD, 50R:50C and high fiber diet (HFD, 60R:40C. Evaluation of salinomycin was carried out using in vitro gas production technique. Methane production and individual fatty acids were estimated by Gas Chromatography. Results: Results of different levels of salinomycin on in vitro methanogenesis indicated that the maximum methane reduction (38.14% in term of mM/gDM was noticed in HFD at 20 ppm level. IVDMD showing increasing trend with an increasing concentration of salinomycin with HFD and LFD, while shown decreasing trend with MFD respectively. Protozoal population significantly decreased by addition of salinomycin in all diets. Conclusion: The results of salinomycin evaluation in the current study can be implicated to mitigate the methane production, thus saving the feed energy loss and the accumulation of green house gases in environment. [Vet World 2012; 5(10.000: 609-613

  9. Comparison of electron beam and gamma ray irradiations effects on ruminal crude protein and amino acid degradation kinetics, and in vitro digestibility of cottonseed meal

    International Nuclear Information System (INIS)

    Ghanbari, F.; Ghoorchi, T.; Shawrang, P.; Mansouri, H.; Torbati-Nejad, N.M.

    2012-01-01

    This study was conducted to compare effects of electron beam (EB) and gamma ray (GR) treatments at doses of 25, 50 and 75 kGy on ruminal degradation kinetics of crude protein (CP), amino acid (AA), and in vitro digestibility of cottonseed meal (CSM). Ionizing radiations of EB and GR had significant effects (P 0.05). Irradiation processing caused decrement in AA degradation after 16 h of ruminal incubation (P<0.05). EB irradiation was more effective than GR irradiation in lessening the ruminal degradability of AA (P<0.05). EB and GR treatments at a dose of 75 kGy increased in vitro digestibility of CSM numerically. This study showed that EB could cause CP and AA bypass rumen as well as GR. Therefore, ionizing irradiation processing can be used as an efficient method in improving nutritional value of CSM. - Highlights: ► Irradiation was effective on reducing ruminal degradability of cottonseed meal. ► Ionizing radiations, especially electron beam, lessened ruminal degradability of amino acid substantially. ► Irradiation processing could be used as a safe and efficient method in improving nutritional value of cottonseed meal.

  10. Accelerated degradation of the D2 protein of photosystem II under ultraviolet radiation

    International Nuclear Information System (INIS)

    Jansen, M.A.K.; Edelman, M.; Greenberg, B.M.; Gaba, V.

    1996-01-01

    The D2 protein of photosystem II is relatively stable in vivo under photosynthetic active radiation, but its degradation accelerates under UVB radiation. Little is known about accelerated D2 protein degradation. We characterized wavelength dependence and sensitivity toward photosystem II inhibitors. The in vivo D2 degradation spectrum resembles the pattern for the rapidly turning over D1 protein of photosystem II, with rates being maximal in the UVB region. We propose that D2 degradation, like D1 degradation, is activated by distinct photosensitizers in the UVB and visible regions of the spectrum. In both wavelength regions, photosystem II inhibitors that are known to be targeted to the D1 protein affect D2 degradation. This suggests that degradation of the two proteins is coupled, D2 degradation being influenced by events occurring at the Q B niche on the D1 protein. (Author)

  11. Protection of protein from ruminal degradation by alkali-induced oxidation of chlorogenic acid in sunflower meal.

    Science.gov (United States)

    Bongartz, V; Böttger, C; Wilhelmy, N; Schulze-Kaysers, N; Südekum, K-H; Schieber, A

    2018-02-01

    Lactating ruminants require an adequate supply of absorbable amino acids for the synthesis of milk protein from two sources, that is crude protein (CP) synthesized microbially in the rumen and ruminally undegraded CP (RUP) from feed which can both be digested in the small intestine. Several chemical and physical methods have been identified as being effective in increasing the proportion of RUP of total CP of a feedstuff, yet there is a continuing need for developing and establishing methods which protect feed protein from ruminal degradation with acceptable expenditure of labour and other costs. The objective of this study was to identify and quantify effects of and interactions between chlorogenic acid and protein in solvent-extracted sunflower meal (SFM) as induced by alkali treatment. Response surface methodology was employed to investigate the influence of pH, reaction time and drying temperature on the resulting SFM and, subsequently, its protein value for ruminants estimated from laboratory values. For this purpose, alkali-treated SFM was subjected to a fractionation of feed CP according to the Cornell net carbohydrate and protein system as a basis for estimating RUP at different assumed ruminal passage rates (K p ). To estimate the intestinal digestibility of the treated SFM and its RUP, a three-step enzymatic in vitro procedure was applied. Alkaline treatment of SFM increased RUP values with factors ranging from approximately 3 (K p =.08/hr) to 12 (K p =.02/hr). Furthermore, the intestinal digestibility of the alkali-treated SFM was enhanced by approximately 10% compared to untreated SFM. Increasing pH and reaction time led to both increasing RUP values and intestinal digestibility. In conclusion, a targeted alkaline treatment of naturally occurring compounds in feedstuffs might be a promising approach to provide high-RUP feeds for ruminants which, at the same time, have improved intestinal digestibility values. © 2017 Blackwell Verlag GmbH.

  12. Secretion of intact proteins and peptide fragments by lysosomal pathways of protein degradation

    International Nuclear Information System (INIS)

    Isenman, L.D.; Dice, J.F.

    1989-01-01

    We report that degradation of proteins microinjected into human fibroblasts is accompanied by release into the culture medium of peptide fragments and intact proteins as well as single amino acids. For the nine proteins and polypeptides microinjected, acid-precipitable radioactivity, i.e. peptide fragments and/or intact proteins, ranged from 10 to 67% of the total released radioactivity. Peptide fragments and/or intact protein accounted for 60% of the radioactivity released into the medium by cells microinjected with ribonuclease A. Two major radiolabeled peptide fragments were found, and one was of an appropriate size to function as an antigen in antigen-presenting cells. The peptides released from microinjected ribonuclease A were derived from lysosomal pathways of proteolysis based on several lines of evidence. Previous studies have shown that microinjected ribonuclease A is degraded to single amino acids entirely within lysosomes. We show that release of free amino acids and peptide fragments and/or intact protein was equivalently stimulated by serum deprivation and equivalently inhibited by NH4Cl. We also show that lysosomal degradation of endocytosed [3H]ribonuclease A was accompanied by the release of two peptide fragments similar in size and charge to those from microinjected [ 3 H]ribonuclease A. These findings demonstrate that degradation within lysosomes occurs in a manner that spares specific peptides; they also suggest a previously unsuspected pathway by which cells can secrete cytosol-derived polypeptides

  13. Abundance and genetic diversity of microbial polygalacturonase and pectate lyase in the sheep rumen ecosystem.

    Directory of Open Access Journals (Sweden)

    Peng Yuan

    Full Text Available Efficient degradation of pectin in the rumen is necessary for plant-based feed utilization. The objective of this study was to characterize the diversity, abundance, and functions of pectinases from microorganisms in the sheep rumen.A total of 103 unique fragments of polygalacturonase (PF00295 and pectate lyase (PF00544 and PF09492 genes were retrieved from microbial DNA in the rumen of a Small Tail Han sheep, and 66% of the sequences of these fragments had low identities (<65% with known sequences. Phylogenetic tree building separated the PF00295, PF00544, and PF09492 sequences into five, three, and three clades, respectively. Cellulolytic and noncellulolytic Butyrivibrio, Prevotella, and Fibrobacter species were the major sources of the pectinases. The two most abundant pectate lyase genes were cloned, and their protein products, expressed in Escherichia coli, were characterized. Both enzymes probably act extracellularly as their nucleotide sequences contained signal sequences, and they had optimal activities at the ruminal physiological temperature and complementary pH-dependent activity profiles.This study reveals the specificity, diversity, and abundance of pectinases in the rumen ecosystem and provides two additional ruminal pectinases for potential industrial use under physiological conditions.

  14. Rumen-protected lysine, methionine, and histidine increase milk protein yield in dairy cows fed a metabolizable protein-deficient diet.

    Science.gov (United States)

    Lee, C; Hristov, A N; Cassidy, T W; Heyler, K S; Lapierre, H; Varga, G A; de Veth, M J; Patton, R A; Parys, C

    2012-10-01

    The objective of this experiment was to evaluate the effect of supplementing a metabolizable protein (MP)-deficient diet with rumen-protected (RP) Lys, Met, and specifically His on dairy cow performance. The experiment was conducted for 12 wk with 48 Holstein cows. Following a 2-wk covariate period, cows were blocked by DIM and milk yield and randomly assigned to 1 of 4 diets, based on corn silage and alfalfa haylage: control, MP-adequate diet (ADMP; MP balance: +9 g/d); MP-deficient diet (DMP; MP balance: -317 g/d); DMP supplemented with RPLys (AminoShure-L, Balchem Corp., New Hampton, NY) and RPMet (Mepron; Evonik Industries AG, Hanau, Germany; DMPLM); and DMPLM supplemented with an experimental RPHis preparation (DMPLMH). The analyzed crude protein content of the ADMP and DMP diets was 15.7 and 13.5 to 13.6%, respectively. The apparent total-tract digestibility of all measured nutrients, plasma urea-N, and urinary N excretion were decreased by the DMP diets compared with ADMP. Milk N secretion as a proportion of N intake was greater for the DMP diets compared with ADMP. Compared with ADMP, dry matter intake (DMI) tended to be lower for DMP, but was similar for DMPLM and DMPLMH (24.5, 23.0, 23.7, and 24.3 kg/d, respectively). Milk yield was decreased by DMP (35.2 kg/d), but was similar to ADMP (38.8 kg/d) for DMPLM and DMPLMH (36.9 and 38.5kg/d, respectively), paralleling the trend in DMI. The National Research Council 2001model underpredicted milk yield of the DMP cows by an average (±SE) of 10.3 ± 0.75 kg/d. Milk fat and true protein content did not differ among treatments, but milk protein yield was increased by DMPLM and DMPLMH compared with DMP and was not different from ADMP. Plasma essential amino acids (AA), Lys, and His were lower for DMP compared with ADMP. Supplementation of the DMP diets with RP AA increased plasma Lys, Met, and His. In conclusion, MP deficiency, approximately 15% below the National Research Council requirements from 2001, decreased

  15. Reticulophagy and Ribophagy: Regulated Degradation of Protein Production Factories

    Directory of Open Access Journals (Sweden)

    Eduardo Cebollero

    2012-01-01

    Full Text Available During autophagy, cytosol, protein aggregates, and organelles are sequestered into double-membrane vesicles called autophagosomes and delivered to the lysosome/vacuole for breakdown and recycling of their basic components. In all eukaryotes this pathway is important for adaptation to stress conditions such as nutrient deprivation, as well as to regulate intracellular homeostasis by adjusting organelle number and clearing damaged structures. For a long time, starvation-induced autophagy has been viewed as a nonselective transport pathway; however, recent studies have revealed that autophagy is able to selectively engulf specific structures, ranging from proteins to entire organelles. In this paper, we discuss recent findings on the mechanisms and physiological implications of two selective types of autophagy: ribophagy, the specific degradation of ribosomes, and reticulophagy, the selective elimination of portions of the ER.

  16. Protein Degradation in Normal and Beige (Chediak-Higashi) Mice

    Science.gov (United States)

    Lyons, Robert T.; Pitot, Henry C.

    1978-01-01

    The beige mouse, C57BL/6 (bg/bg), is an animal model for the Chediak-Higashi syndrome in man, a disease characterized morphologically by giant lysosomes in most cell types. Half-lives for the turnover of [14C]bicarbonate-labeled total soluble liver protein were determined in normal and beige mice. No significant differences were observed between the normal and mutant strain for both rapidly and slowly turning-over classes of proteins. Glucagon treatment during the time-course of protein degradation had similar effects on both normal and mutant strains and led to the conclusion that the rate of turnover of endogenous intracellular protein in the beige mouse liver does not differ from normal. The rates of uptake and degradation of an exogenous protein were determined in normal and beige mice by intravenously injecting 125I-bovine serum albumin and following, in peripheral blood, the loss with time of phosphotungstic acid-insoluble bovine serum albumin and the parallel appearance of phosphotungstic acid-soluble (degraded) material. No significant differences were observed between beige and normal mice in the uptake by liver lysosomes of 125I-bovine serum albumin (t½ = 3.9 and 2.8 h, respectively). However, it was found that lysosomes from livers of beige mice released phosphotungstic acid-soluble radioactivity at a rate significantly slower than normal (t½ = 6.8 and 3.1 h, respectively). This defect in beige mice could be corrected by chronic administration of carbamyl choline (t½ = 3.5 h), a cholinergic agonist which raises intracellular cyclic GMP levels. However, no significant differences between normal and beige mice were observed either in the ability of soluble extracts of liver and kidney to bind [3H]cyclic GMP in vitro or in the basal levels of cyclic AMP in both tissues. The relevance of these observations to the presumed biochemical defect underlying the Chediak-Higashi syndrome is discussed. PMID:202611

  17. Profile of Rumen Fermentation and Blood Urea Nitrogen Concentration of Kacang Goat Fed Total Mixed Ration Vs. Roughage

    Science.gov (United States)

    Adiwinarti, R.; Kustantinah; Budisatria, I. G. S.; Rusman; Indarto, E.

    2018-02-01

    Kacang goat is usually reared traditionally fed natural grass and having inferior performance. Many researches had been done to improve the performance. Total mixed ration (TMR) containing soybean meal (SBM) and fish meal (FM) could increase the performance of Kacang goat, but the profile of rumen fermentation has not been published. Therefore, this study investigated the profile of rumen fermentation and blood urea Nitrogen concentration (BUN) in Kacang goat fed natural grass (roughage) and TMR. Twelve yearling Kacang buck, 15.2-19.6 kg were arranged in completely randomized design. The treatments were NG (natural grass), FM (TMR containing FM), and SBM (TMR containing SBM). The TMR were formulated containing 15% crude protein and 56-58% TDN. Data were analyzed by one way ANOVA. Rumen pH 6 hours after feeding of NG (7.4) was higher (P<0.01) than that of FM (6.2) and SBM (6.4). This lowering pH of TMR was caused by increasing volatile fatty acids (VFA). The VFA total of FM (129.7 mmol/l) and SBM (153.1 mmol/l) were higher than that of NG (86.4 mmol/l). At 3 and 6 hours after feeding, ammonia in the rumen of SBM was higher than that of NG and FM, indicating higher degraded protein. The BUN at 3 hours after feeding of SBM was higher than that of NG. It can be concluded that protein in SBM was degraded higher than others and the lower pH in rumen of TMR goats was caused by higher VFA produced by TMR goats compared to NG goats.

  18. Symbiosis and Rumen Protozoa

    Science.gov (United States)

    Dillon, Raymond D.

    1970-01-01

    Protozoa inhabiting the rumen of large grazing animals can be used to illustrate symbiotic animal associations. Gives a key to the ciliates most commonly found, several drawings, and a chart relating rumen fauna to the phylogenetic tree of the hosts. (EB)

  19. Evaluation of the efficacy of rumen cannulation technique on some rumen metabolic parameters in buffaloes

    Directory of Open Access Journals (Sweden)

    Gh. Manafiazar

    2010-02-01

    Full Text Available This experimental study was carried out to determine the efficacy of rumen cannulation technique on some rumen metabolic parameters in buffaloes. Four healthy male River (Azari buffaloes with no previous history of gastrointestinal dysfunction were chosen. There are several surgical techniques that can be used for rumen cannulation in farm animals, including buffaloes. This procedure was performed in a modified two-stage technique. In the first stage, the dorso – lateral portion of the dorsal sac of the rumen wall was sutured to the skin incision in the left para lumbar fossa region. In the second – stage, after six days left, the exposed rumen wall area was incised and the cannula was inserted and fixed manually in the rumenal opening incision site. In order to evaluate the efficacy of rumen cannulation technique on some rumen metabolic parameters, this study was achieved with different levels of NDF, and chewing behavior and their relationship with ruminal acidity, was measured in a change over design. Two diets with 2 levels of NDF were used as treatments. First and second diets had 52 and 47 % of NDF, respectively. Animals fed ad libitum at 09:00 and 21:00. There were no significant differences between chemical composition, particles distribution, geometric mean, its standard deviation and physically effective factor (pef of diets, dry matter intake (kg/d and nutrients intake (NDF, ADF, NFC and crude protein and their digestibility. Increasing NFC reduced ruminal pH at 0.5, 1.0, 4.5, 6.0 9.0 and 10.0 h post feeding. In addition, there were not significant differences on eating time, rumination time and total chewing activity between diets. All data obtained in this study were in normal range may indicating the efficacy of this cannulation method. More investigation should be done to determine the efficacy and comparison of the other surgical rumen cannulation techniques on Azari buffaloes of Iran.

  20. Study on the rumen degradability of six types of common roughage in Tongliao Khorchin beef cattle%通辽地区科尔沁肉牛6种常用粗饲料瘤胃降解特性的研究

    Institute of Scientific and Technical Information of China (English)

    张颖; 接彩虹

    2018-01-01

    为了研究科尔沁肉牛对不同粗饲料的降解特性,采用尼龙袋法测定苜蓿干草、玉米秸秆、羊草、稻草、玉米青贮和苜蓿草块6种粗饲料在肉牛瘤胃内干物质(dry matter,DM)、粗蛋白(crude protein,CP)、中性洗涤纤维(neutral detergent fiber,NDF)和酸性洗涤纤维(acid detergent fiber,ADF)的瘤胃降解率.结果显示:DM和CP的肉牛瘤胃有效降解率(ED)以苜蓿干草最高,稻草最低,苜蓿草块、玉米青贮、羊草和玉米秸秆居中;NDF和ADF的肉牛瘤胃ED以苜蓿干草最高,羊草最低.结果表明:从科尔沁肉牛对6种粗饲料的降解特性看,苜蓿干草的营养价值最高,稻草的营养价值最低.%To study the degradation characteristics of different types of roughage in Khorchin Beef Cattle,the nylon bag method was used to determine the ruminal degradability of six types of roughage:dry matter (DM),crude protein (CP),neutral detergent fiber (NDF) and acid detergent fiber (ADF).The results were as follows:of the six types of roughage,alfalfa hay was of the most effective DM and CP degradability in rumen;rice straw was the least effective;and alfalfa block,corn silage,leymus chinensis and corn stover were in the middle of the scale of effective DM and CP degradability.In terms of NDF and ADF degradability,alfalfa hay was the most effective,while leymus chinensis was the least effective.In conclusion,in consideration of the degradation characteristics of six types of roughage in Khorchin beef cattle,alfalfa hay possesses the highest nutritional value,while rice straw has the lowest value.

  1. Mass spectrometry analysis of proteome-wide proteolytic post-translational degradation of proteins

    OpenAIRE

    Shen, Yufeng; Hixson, Kim K.; Tolić, Nikola; Camp, David G.; Purvine, Samuel O.; Moore, Ronald J.; Smith, Richard D.

    2008-01-01

    Protein proteolytic degradation is an essential component to proper cell function and its life cycle. Here, we study the protein degradation in yeast Saccharomyces cerevisiae cells on a proteome-wide scale by detection of the intermediate peptides produced from the intracellular degradation of proteins using sequencing-based tandem mass spectrometry. By tracing the detected ~1,100 peptides and their ~200 protein substrate origins we obtain evidence for new insights into the proteome-wide prot...

  2. Effect of supplementation of concentrates or selenium on production and reproduction in cows grazing pastures of high protein degradability

    Energy Technology Data Exchange (ETDEWEB)

    Mongiardino, M E; Humaran, M; Corbellini, C N; Baldan, A M; Cuneo, M; Balbiani, G [Instituto Nacional de Tecnologia Agropecuaria, Moron, Provincia de Buenos Aires (Argentina). Inst. de Patobiologia

    1996-05-01

    Two experiments were carried out to determine whether the deleterious effects of high amounts of degradable protein on reproduction and production of dairy cows could be minimized by a supplemental source of undergradable protein, or grain supplementation while grazing; and to study the effect of selenium supplementation before calving on the incidence of stillbirths, mastitis, puerperal and metabolic disorders. In a first experiment, 24 Holstein cows fed on red and white clover pasture, paired by previous milk production, calving data and body condition, were supplemented with corn silage and one of two concentrates differing only in the proportion of degradable protein (Group H: 71.5% and Group L: 51.5%). The degradable protein intake from pasture supplied 93% of the requirements in both groups. The addition of undergradable protein in the concentrate of Group L did not improve reproductive performance nor milk, butterfat or solids non-fat production. In a second experiment, 132 Holstein cows and heifers were paired likewise. Both animals in each pair were fed similar forage resources, but each one was supplemented with 2 kg/cow corn grain four times a day (Herd 1) or 4 kg/cow tow times a day (Herd 2). One animal in each pair was randomly assigned to receive a barium selenate injection before calving. Rumen ammonia was higher in Herd 1 in both sampling dates (17 vs 4.2 mg/100 ml and 12 vs 9 mg/100 ml), as well as serum urea up to 50 days post-partum (26 vs 19 mg/100 ml, P<0.02). Body condition scores were similar at calving but significantly lower in Herd 1 during the lactation period (P<0.05). Total milk and butterfat production were higher in Herd 2 (6406.2 vs 6893.8 kg and 190.4 vs 203.5 kg, respectively). Selenium improved pregnancy rate to first artificial insemination in Herd 2 (71 vs 50%), and decreased the frequency of downer cows in both herds (5 vs 0%), but had not effect regarding the Wisconsin Mastitis Test results. (author). 36 refs, 4 figs, 3 tabs.

  3. Synthesis of microbial nitrogen compounds in the rumen and their digestion in the small intestine

    International Nuclear Information System (INIS)

    Smith, R.H.; Salter, D.N.; McAllan, A.B.; Williams, A.P.

    1976-01-01

    Pathways and efficiency of NPN utilization were studied after introducing isonitrogenous feeds containing urea labelled with 15 N with or without groundnut protein into the rumen of the young steer. The 15 N abundance in rumen ammonia decreased from 1 h after the dose whereas that in suspended bacteria (only a small part of the total population but regarded, with reservation, as representative) increased to a peak value at 3-5 h. Patterns of incorporation into different nitrogen constituents of the bacteria indicated that part of the 15 N first moved rapidly into a labile pool, which included amide nitrogen, and then moved from this pool into other amino acids. Different amino acids were labelled to different extents (lowest for proline and arginine, highest for aspartic acid and alanine), presumably as different amounts of pre-formed amino acids were used in protein synthesis. Differences largely disappeared when the diet with urea as the only nitrogen source was given. Estimates of microbial 15 N recovery at the duodenum confirmed earlier findings that overall incorporation was least when the latter diet was given, and no improvement was seen when the urea was given in three doses over 4 h instead of in a single dose. Lactosyl, like glucosyl, ureide was degraded fairly slowly in the rumen and is being studied as a possible NPN source likely to yield simultaneous ammonia and energy for microbial growth. Studies on the origin and composition of nitrogen compounds entering the duodenum showed that the RNA of rumen bacteria (labelled with 32 P) contributed 82-109% of the RNA entering the duodenum of the steer. It was also shown that protecting dietary casein with formaldehyde reduced its degradability in the rumen to such an extent that a deficiency of fermentable nitrogen in that organ ensued. In other experiments with two calves receiving flaked maize and hay, net digestibility of nitrogen compounds between duodenum and ileum was about 60%. True digestibility

  4. Effects of Protein Level and Mangosteen Peel Pellets (Mago-pel in Concentrate Diets on Rumen Fermentation and Milk Production in Lactating Dairy Crossbreds

    Directory of Open Access Journals (Sweden)

    T. Norrapoke

    2012-07-01

    Full Text Available Four, lactating dairy crossbreds (50%×50% Holstein Friesian×Native Zebu cattle were randomly assigned according to a 2×2 factorial arrangement (two protein levels and two levels of mangosteen peel pellets (Mago-pel in a 4×4 Latin square design to receive four dietary treatments. All cows received concentrate at a proportion of 1 kg concentrate per 2 kg of milk yield, and urea-treated 5% rice straw (UTRS was given ad libitum. It was found that total dry matter intakes, nutrient digestibility, ruminal pH and NH3-N concentrations were not affected (p>0.05 by treatments. Concentrations of ruminal pH and NH3-N were not affected by dietary treatments although the concentration of BUN varied significantly (p0.05; however, the population of protozoa was decreased (p0.05 among dietary treatments (p>0.05; however, copy numbers of Ruminococcus flavefaciens was increased when protein level increased (p0.05 milk composition except solids-not-fat which was higher in cows fed the diet with 19% CP. Therefore, feeding a concentrate containing 16% CP together with 300 g/hd/d Mago-pel supplementation results in changes in rumen fermentation and microbial population and improvements in milk production in lactating dairy crossbreds fed on UTRS.

  5. Effect of the Ratio of Non-fibrous Carbohydrates to Neutral Detergent Fiber and Protein Structure on Intake, Digestibility, Rumen Fermentation, and Nitrogen Metabolism in Lambs.

    Science.gov (United States)

    Ma, T; Tu, Y; Zhang, N F; Deng, K D; Diao, Q Y

    2015-10-01

    This study aimed to investigate the effect of the ratio of non-fibrous carbohydrates to neutral detergent fibre (NFC/NDF) and undegraded dietary protein (UDP) on rumen fermentation and nitrogen metabolism in lambs. Four Dorper×thin-tailed Han crossbred lambs, averaging 62.3±1.9 kg of body weight and 10 mo of age, were randomly assigned to four dietary treatments of combinations of two levels of NFC/NDF (1.0 and 1.7) and two levels of UDP (35% and 50% of crude protein [CP]). Duodenal nutrient flows were measured with dual markers of Yb and Co, and microbial N (MN) synthesis was estimated using (15)N. High UDP decreased organic matter (OM) intake (p = 0.002) and CP intake (p = 0.005). Ruminal pH (p<0.001), ammonia nitrogen (NH3-N; p = 0.008), and total volatile fatty acids (p<0.001) were affected by dietary NFC/NDF. The ruminal concentration of NH3-N was also affected by UDP (p<0.001). The duodenal flow of total MN (p = 0.007) was greater for lambs fed the high NFC/NDF diet. The amount of metabolisable N increased with increasing dietary NFC:NDF (p = 0.02) or UDP (p = 0.04). In conclusion, the diets with high NFC/NDF (1.7) and UDP (50% of CP) improved metabolisable N supply to lambs.

  6. Effect of the Ratio of Non-fibrous Carbohydrates to Neutral Detergent Fiber and Protein Structure on Intake, Digestibility, Rumen Fermentation, and Nitrogen Metabolism in Lambs

    Directory of Open Access Journals (Sweden)

    T. Ma

    2015-10-01

    Full Text Available This study aimed to investigate the effect of the ratio of non-fibrous carbohydrates to neutral detergent fibre (NFC/NDF and undegraded dietary protein (UDP on rumen fermentation and nitrogen metabolism in lambs. Four Dorper×thin-tailed Han crossbred lambs, averaging 62.3±1.9 kg of body weight and 10 mo of age, were randomly assigned to four dietary treatments of combinations of two levels of NFC/NDF (1.0 and 1.7 and two levels of UDP (35% and 50% of crude protein [CP]. Duodenal nutrient flows were measured with dual markers of Yb and Co, and microbial N (MN synthesis was estimated using 15N. High UDP decreased organic matter (OM intake (p = 0.002 and CP intake (p = 0.005. Ruminal pH (p<0.001, ammonia nitrogen (NH3-N; p = 0.008, and total volatile fatty acids (p<0.001 were affected by dietary NFC/NDF. The ruminal concentration of NH3-N was also affected by UDP (p<0.001. The duodenal flow of total MN (p = 0.007 was greater for lambs fed the high NFC/NDF diet. The amount of metabolisable N increased with increasing dietary NFC:NDF (p = 0.02 or UDP (p = 0.04. In conclusion, the diets with high NFC/NDF (1.7 and UDP (50% of CP improved metabolisable N supply to lambs.

  7. Live yeast culture and monensin in high grain diets for cattle: rumen fermentation and “in situ” degradability Leveduras vivas e monensina em dietas de alto concentrado para bovinos: parâmetros ruminais e degradabilidade "in situ"

    Directory of Open Access Journals (Sweden)

    José Carlos Machado Nogueira Filho

    2010-03-01

    Full Text Available Live yeast cultures have been used as an alternative to replace antibiotics in diets for ruminants. Therefore, the aim was to evaluate the effects of adding live yeast culture (Saccharomyces cerevisiae, Beef Sacc®, Alltech, Inc., monensin (Rumensin®, Elanco, Inc. and the combination of both additives in high grain diets, on rumen fermentation patterns, protozoa organisms and in situ degradability of diet components. Four rumen-cannulated steers were fed a basal ration (2.8Mcal ME/kg DM, 14% CP and submitted to one of four treatments following a 4x4 Latin Square design: control (CON, no additives, yeast (YEA, 0.6g/kg of dry matter, monensin (MON, 0.3g/kg of dry matter and monensin plus yeast (MON+YEA. After 14 days of diet adaptation, the rumen fermentation parameters, the protozoa numbers and the degradation kinetics of corn (CO, soybean meal (SM and soybean hulls (SH were assessed. Feed additives did not affect rumen pH, butirate and ammonia nitrogen concentrations, but decreased total short chain fatty acids (mM. MON and MON+ YEA decreased acetate (% and acetate:propionate ratio whereas propionate was increased by MON and MON+YEA at all sampling times, and by YEA at 4 and 6h post-feeding only. YEA increased the number of protozoa whereas MON and MON+YEA inhibited those microorganisms (total organisms [x104/mL]. There were no effects of YEA and MON+YEA on in situ degradability parameters of any evaluated feed, however, MON increased NDF degradation rate of SH. Monensin effects on rumen fermentation are more significant than those observed when feeding live yeast cultures, and the combination of both additives does not improve their effects.Leveduras têm sido utilizadas para substituir antibióticos em dietas para ruminantes. Assim, objetivou-se avaliar o efeito de leveduras vivas (Saccharomyces cerevisiae, Beef-Sacc®, Alltech, Inc., monensina (Rumensin®, Elanco, Inc. e a combinação de ambos em dietas com alto concentrado, sobre a

  8. Induced oligomerization targets Golgi proteins for degradation in lysosomes.

    Science.gov (United States)

    Tewari, Ritika; Bachert, Collin; Linstedt, Adam D

    2015-12-01

    Manganese protects cells against forms of Shiga toxin by down-regulating the cycling Golgi protein GPP130. Down-regulation occurs when Mn binding causes GPP130 to oligomerize and traffic to lysosomes. To determine how GPP130 is redirected to lysosomes, we tested the role of GGA1 and clathrin, which mediate sorting in the canonical Golgi-to-lysosome pathway. GPP130 oligomerization was induced using either Mn or a self-interacting version of the FKBP domain. Inhibition of GGA1 or clathrin specifically blocked GPP130 redistribution, suggesting recognition of the aggregated GPP130 by the GGA1/clathrin-sorting complex. Unexpectedly, however, GPP130's cytoplasmic domain was not required, and redistribution also occurred after removal of GPP130 sequences needed for its normal cycling. Therefore, to test whether aggregate recognition might be a general phenomenon rather than one involving a specific GPP130 determinant, we induced homo-oligomerization of two unrelated Golgi-targeted constructs using the FKBP strategy. These were targeted to the cis- and trans-Golgi, respectively, using domains from mannosidase-1 and galactosyltransferase. Significantly, upon oligomerization, each redistributed to peripheral punctae and was degraded. This occurred in the absence of detectable UPR activation. These findings suggest the unexpected presence of quality control in the Golgi that recognizes aggregated Golgi proteins and targets them for degradation in lysosomes. © 2015 Tewari et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  9. Effects of gas composition in headspace and bicarbonate concentrations in media on gas and methane production, degradability, and rumen fermentation using in vitro gas production techniques.

    Science.gov (United States)

    Patra, Amlan Kumar; Yu, Zhongtang

    2013-07-01

    Headspace gas composition and bicarbonate concentrations in media can affect methane production and other characteristics of rumen fermentation in in vitro gas production systems, but these 2 important factors have not been evaluated systematically. In this study, these 2 factors were investigated with respect to gas and methane production, in vitro digestibility of feed substrate, and volatile fatty acid (VFA) profile using in vitro gas production techniques. Three headspace gas compositions (N2+ CO2+ H2 in the ratio of 90:5:5, CO2, and N2) with 2 substrate types (alfalfa hay only, and alfalfa hay and a concentrate mixture in a 50:50 ratio) in a 3×2 factorial design (experiment 1) and 3 headspace compositions (N2, N2 + CO2 in a 50:50 ratio, and CO2) with 3 bicarbonate concentrations (80, 100, and 120 mM) in a 3×3 factorial design (experiment 2) were evaluated. In experiment 1, total gas production (TGP) and net gas production (NGP) was the lowest for CO2, followed by N2, and then the gas mixture. Methane concentration in headspace gas after fermentation was greater for CO2 than for N2 and the gas mixture, whereas total methane production (TMP) and net methane production (NMP) were the greatest for CO2, followed by the gas mixture, and then N2. Headspace composition did not affect in vitro digestibility or the VFA profile, except molar percentages of propionate, which were greater for CO2 and N2 than for the gas mixture. Methane concentration in headspace gas, TGP, and NGP were affected by the interaction of headspace gas composition and substrate type. In experiment 2, increasing concentrations of CO2 in the headspace decreased TGP and NGP quadratically, but increased the concentrations of methane, NMP, and in vitro fiber digestibility linearly, and TMP quadratically. Fiber digestibility, TGP, and NGP increased linearly with increasing bicarbonate concentrations in the medium. Concentrations of methane and NMP were unaffected by bicarbonate concentration, but

  10. Critical evaluation of essential oils as rumen modifiers in ruminant nutrition: A review

    Energy Technology Data Exchange (ETDEWEB)

    Cobellis, Gabriella, E-mail: cobellis.gabriella@gmail.com [Dipartimento di Medicina Veterinaria, Università degli Studi di Perugia, Perugia (Italy); Department of Animal Sciences, The Ohio State University, Columbus, OH 43210 (United States); Trabalza-Marinucci, Massimo [Dipartimento di Medicina Veterinaria, Università degli Studi di Perugia, Perugia (Italy); Yu, Zhongtang [Department of Animal Sciences, The Ohio State University, Columbus, OH 43210 (United States)

    2016-03-01

    Ruminant livestock systems contribute significantly to emission of methane, a potent greenhouse gas as they waste a portion of the ingested energy (2–15%) as methane and a large proportion (75–95%) of the ingested nitrogen as ammonia. Recently, numerous researches have been conducted to evaluate plant secondary metabolites, including essential oils (EO), as natural feed additives in ruminant nutrition and to exploit their potential to improve rumen fermentation efficiency. Essential oils appeared to be very promising compounds as they selectively reduced methane production and protein breakdown in both in vitro and in vivo studies. However, in some studies, the use of EO as feed additives was accompanied with decreased feed degradability and lowered volatile fatty acid. These adverse effects could be attributed to their broad and often non-specific antimicrobial activities within the rumen. Future research should be directed to identification of the active and useful EO compounds, optimization of EO doses, and use of a whole-farm approach with a focus on animal welfare, performance and economic benefits. - Highlights: • Ruminants contributes 16–25% to the global greenhouse gases emissions. • Decrease methane emission and nitrogen excretion from ruminant livestock industry is urgently needed. • Essential oils have been shown to be promising feed additives in mitigating methane and ammonia emissions. • Essential oils have showed inconsistent results about feed degradability and VFA production. • The mode of action and activities of essential oils on rumen microbiome remain poorly understood.

  11. Critical evaluation of essential oils as rumen modifiers in ruminant nutrition: A review

    International Nuclear Information System (INIS)

    Cobellis, Gabriella; Trabalza-Marinucci, Massimo; Yu, Zhongtang

    2016-01-01

    Ruminant livestock systems contribute significantly to emission of methane, a potent greenhouse gas as they waste a portion of the ingested energy (2–15%) as methane and a large proportion (75–95%) of the ingested nitrogen as ammonia. Recently, numerous researches have been conducted to evaluate plant secondary metabolites, including essential oils (EO), as natural feed additives in ruminant nutrition and to exploit their potential to improve rumen fermentation efficiency. Essential oils appeared to be very promising compounds as they selectively reduced methane production and protein breakdown in both in vitro and in vivo studies. However, in some studies, the use of EO as feed additives was accompanied with decreased feed degradability and lowered volatile fatty acid. These adverse effects could be attributed to their broad and often non-specific antimicrobial activities within the rumen. Future research should be directed to identification of the active and useful EO compounds, optimization of EO doses, and use of a whole-farm approach with a focus on animal welfare, performance and economic benefits. - Highlights: • Ruminants contributes 16–25% to the global greenhouse gases emissions. • Decrease methane emission and nitrogen excretion from ruminant livestock industry is urgently needed. • Essential oils have been shown to be promising feed additives in mitigating methane and ammonia emissions. • Essential oils have showed inconsistent results about feed degradability and VFA production. • The mode of action and activities of essential oils on rumen microbiome remain poorly understood.

  12. Rapid degradation of abnormal proteins in vacuoles from Acer pseudoplatanus L. cells

    International Nuclear Information System (INIS)

    Canut, H.; Alibert, G.; Carrasco, A.; Boudet, A.M.

    1986-01-01

    In Acer pseudoplatanus cells, the proteins synthesized in the presence of an amino acid analog ([ 14 C]p-fluorophenylalanine), were degraded more rapidly than normal ones ([ 14 C]phenylalanine as precursor). The degradation of an important part of these abnormal proteins occurred inside the vacuoles. The degradation process was not apparently associated to a specific proteolytic system but was related to a preferential transfer of these aberrant proteins from the cytoplasm to the vacuole

  13. Common Prairie feeds with different soluble and insoluble fractions used for CPM diet formulation in dairy cattle: Impact of carbohydrate-protein matrix structure on protein and other primary nutrient digestion

    Science.gov (United States)

    Peng, Quanhui; Wang, Zhisheng; Zhang, Xuewei; Yu, Peiqiang

    2014-03-01

    An experiment was conducted to investigate the relationship of carbohydrates molecular spectral characteristics to rumen degradability of primary nutrients in Prairie feeds in dairy cattle. In total, 12 different types of feeds were selected, each type of feed was from three different source with total 37 samples. Six types of them were energy-sourced feeds and the others were protein-sourced feeds. The carbohydrates molecular spectral intensity of various functional groups were collected using Fourier transform infrared attenuated total reflectance (ATR-FT/IR) spectroscopy. In the in situ study, the results showed that the rumen digestibility and digestible fractions of primary nutrients (DM, OM, NCP, and CP) were significantly different (P digestibility and digestible fractions of DM, OM and NCP. Spectral intensities of H_1150, H_1015, A_1, and A_3 were weakly negatively associated with in situ rumen degradation of CP. Spectral intensities of A_1240 and H_1240, mainly associated with cellulosic compounds, were correlated with rumen CP degradation. The multiple regression analysis demonstrated that the spectral intensities of A_3 and H_1415 played the most important role and could be used as a potential tool to predict rumen protein degradation of feeds in dairy cattle. In conclusion, this study showed that the carbohydrates as a whole have an effect on protein rumen degradation, rather than cellulose alone, indicating carbohydrate-protein matrix structure impact protein utilization in dairy cattle. The non-invasive molecular spectral technique (ATR-FT/IR) could be used as a rapid potential tool to predict rumen protein degradation of feedstuffs by using molecular spectral bands intensities in carbohydrate fingerprint region.

  14. Nanobody-Directed Specific Degradation of Proteins by the 26S-Proteasome in Plants

    OpenAIRE

    Baudisch, Bianca; Pfort, Ingrid; Sorge, Eberhard; Conrad, Udo

    2018-01-01

    Here, we present data showing the directed degradation of target proteins recognized by a specific nanobody in transgenic plants. Green fluorescent protein was depleted by a chimeric nanobody fused to a distinct F-box domain, which enables protein degradation via the ubiquitin proteasome pathway. This technique could thus be used to knock out other proteins of interest in planta using specific, high-affinity binding proteins.

  15. Nanobody-Directed Specific Degradation of Proteins by the 26S-Proteasome in Plants

    Directory of Open Access Journals (Sweden)

    Bianca Baudisch

    2018-02-01

    Full Text Available Here, we present data showing the directed degradation of target proteins recognized by a specific nanobody in transgenic plants. Green fluorescent protein was depleted by a chimeric nanobody fused to a distinct F-box domain, which enables protein degradation via the ubiquitin proteasome pathway. This technique could thus be used to knock out other proteins of interest in planta using specific, high-affinity binding proteins.

  16. Nanobody-Directed Specific Degradation of Proteins by the 26S-Proteasome in Plants.

    Science.gov (United States)

    Baudisch, Bianca; Pfort, Ingrid; Sorge, Eberhard; Conrad, Udo

    2018-01-01

    Here, we present data showing the directed degradation of target proteins recognized by a specific nanobody in transgenic plants. Green fluorescent protein was depleted by a chimeric nanobody fused to a distinct F-box domain, which enables protein degradation via the ubiquitin proteasome pathway. This technique could thus be used to knock out other proteins of interest in planta using specific, high-affinity binding proteins.

  17. In vitro digestibility of individual amino acids in rumen-undegraded protein: the modified three-step procedure and the immobilized digestive enzyme assay.

    Science.gov (United States)

    Boucher, S E; Calsamiglia, S; Parsons, C M; Stern, M D; Moreno, M Ruiz; Vázquez-Añón, M; Schwab, C G

    2009-08-01

    Three soybean meal, 3 SoyPlus (West Central Cooperative, Ralston, IA), 5 distillers dried grains with solubles, and 5 fish meal samples were used to evaluate the modified 3-step in vitro procedure (TSP) and the in vitro immobilized digestive enzyme assay (IDEA; Novus International Inc., St. Louis, MO) for estimating digestibility of AA in rumen-undegraded protein (RUP-AA). In a previous experiment, each sample was ruminally incubated in situ for 16 h, and in vivo digestibility of AA in the intact samples and in the rumen-undegraded residues (RUR) was obtained for all samples using the precision-fed cecectomized rooster assay. For the modified TSP, 5 g of RUR was weighed into polyester bags, which were then heat-sealed and placed into Daisy(II) incubator bottles. Samples were incubated in a pepsin/HCl solution followed by incubation in a pancreatin solution. After this incubation, residues remaining in the bags were analyzed for AA, and digestibility of RUP-AA was calculated based on disappearance from the bags. In vitro RUP-AA digestibility estimates obtained with this procedure were highly correlated to in vivo estimates. Corresponding intact feeds were also analyzed via the pepsin/pancreatin steps of the modified TSP. In vitro estimates of AA digestibility of the feeds were highly correlated to in vivo RUP-AA digestibility, which suggests that the feeds may not need to be ruminally incubated before determining RUP-AA digestibility in vitro. The RUR were also analyzed via the IDEA kits. The IDEA values of the RUR were good predictors of RUP-AA digestibility in soybean meal, SoyPlus, and distillers dried grains with solubles, but the IDEA values were not as good predictors of RUP-AA digestibility in fish meal. However, the IDEA values of intact feed samples were also determined and were highly correlated to in vivo RUP-AA digestibility for all feed types, suggesting that the IDEA value of intact feeds may be a better predictor of RUP-AA digestibility than the IDEA

  18. Effects of different sources of protein on digestive characteristics, microbial efficiency, and nutrient flow in dairy goats

    Directory of Open Access Journals (Sweden)

    Nivea Regina de Oliveira Felisberto

    2011-10-01

    Full Text Available Diets formulated with protein sources presenting different resistance to ruminal degradation were compared by evaluating ruminal parameters, production and microbial efficiency and nutrients flow to the omasum in goats. Eight rumen cannulated non-lactating, non-pregnant goats were distributed in a 4 × 4 Latin square design with two replicates. Treatments consisted of four diets where different sources of plant protein accounted for the major protein source named soybean meal, source of higher ruminal degradability, and three other sources of higher resistance of degradation: roasted soybean, corn gluten meal, and cottonseed cake. Amounts of rumen protein were similar among rations; however, flows of dry matter, protein and non-fiber carbohydrate to omasum were higher for diets with protein source with reduced rumen degradation rate. Higher values of rumen ammonia were obtained by using ration with soybean meal as major source of protein. Higher values of pH were obtained for rations with roasted soybean e cottonseed cake. Regarding kinetic of transit, similar values were found among rations. Diets with protein sources presenting reduced ruminal degradation increase nutrients flow to the omasum in goats and alter digestive parameters such as pH and ammonia without compromising bacteria growth and efficiency, which grants their use for dairy goats with similar efficiency to rations using more degradable sources of protein.

  19. Diversity of phytases in the rumen.

    Science.gov (United States)

    Nakashima, Brenda A; McAllister, Tim A; Sharma, Ranjana; Selinger, L Brent

    2007-01-01

    Examples of a new class of phytase related to protein tyrosine phosphatases (PTP) were recently isolated from several anaerobic bacteria from the rumen of cattle. In this study, the diversity of PTP-like phytase gene sequences in the rumen was surveyed by using the polymerase chain reaction (PCR). Two sets of degenerate primers were used to amplify sequences from rumen fluid total community DNA and genomic DNA from nine bacterial isolates. Four novel PTP-like phytase sequences were retrieved from rumen fluid, whereas all nine of the anaerobic bacterial isolates investigated in this work contained PTP-like phytase sequences. One isolate, Selenomonas lacticifex, contained two distinct PTP-like phytase sequences, suggesting that multiple phytate hydrolyzing enzymes are present in this bacterium. The degenerate primer and PCR conditions described here, as well as novel sequences obtained in this study, will provide a valuable resource for future studies on this new class of phytase. The observed diversity of microbial phytases in the rumen may account for the ability of ruminants to derive a significant proportion of their phosphorus requirements from phytate.

  20. Influence of anabolic agents on protein synthesis and degradation in muscle cells grown in culture

    Energy Technology Data Exchange (ETDEWEB)

    Roeder, R.A.; Thorpe, S.D.; Byers, F.M.; Schelling, G.T.; Gunn, J.M.

    Muscle cell culture (L/sub 6/) studies were conducted to determine whether anabolic agents have a direct effect on the muscle cell. The effect of zeranol, testosterone propionate, estradiol benzoate, progesterone, dexamethasone and anabolic agent-dexamethasone combinations on protein synthesis and degradation were measured. Myoblast and myotube cultures were pretreated with 1 ..mu..M compounds for 12, 24 and 48 h before a 6-h synthesis or degradation measuring period. Protein synthesis was determined as cpm of (/sup 3/H) leucine incorporated per mg cell protein. Protein degradation was measured by a pulse-chase procedure using (/sup 3/H) leucine and expressed as the percentage labeled protein degraded in 6 h. Progesterone slightly increased protein synthesis in myoblast cultures. Testosterone propionate had no effect on synthesis. Protein synthesis was decreased by estradiol benzoate in myotube cultures. Protein degradation was not altered appreciably by anabolic agents. Protein synthesis was initially inhibited in myotubes by dexamethasone, but increased in myoblasts and myotubes in the extended incubation time. Dexamethasone also consistently increased protein degradation, but this required several hours to be expressed. Anabolic agents did not interfere with dexamethasone-induced increases in protein synthesis and degradation. The magnitude of response and sensitivity were similar for both the myoblast and the more fully differentiated myotube for all compounds tested. These results indicate that anabolic agents at the 1 ..mu..M level do not have a direct anabolic effect on muscle or alter glucocorticoid-induced catabolic response in muscle.

  1. Influence of anabolic agents on protein synthesis and degradation in muscle cells grown in culture

    International Nuclear Information System (INIS)

    Roeder, R.A.; Thorpe, S.D.; Byers, F.M.; Schelling, G.T.; Gunn, J.M.

    1986-01-01

    Muscle cell culture (L 6 ) studies were conducted to determine whether anabolic agents have a direct effect on the muscle cell. The effect of zeranol, testosterone propionate, estradiol benzoate, progesterone, dexamethasone and anabolic agent-dexamethasone combinations on protein synthesis and degradation were measured. Myoblast and myotube cultures were pretreated with 1 μM compounds for 12, 24 and 48 h before a 6-h synthesis or degradation measuring period. Protein synthesis was determined as cpm of [ 3 H] leucine incorporated per mg cell protein. Protein degradation was measured by a pulse-chase procedure using [ 3 H] leucine and expressed as the percentage labeled protein degraded in 6 h. Progesterone slightly increased protein synthesis in myoblast cultures. Testosterone propionate had no effect on synthesis. Protein synthesis was decreased by estradiol benzoate in myotube cultures. Protein degradation was not altered appreciably by anabolic agents. Protein synthesis was initially inhibited in myotubes by dexamethasone, but increased in myoblasts and myotubes in the extended incubation time. Dexamethasone also consistently increased protein degradation, but this required several hours to be expressed. Anabolic agents did not interfere with dexamethasone-induced increases in protein synthesis and degradation. The magnitude of response and sensitivity were similar for both the myoblast and the more fully differentiated myotube for all compounds tested. These results indicate that anabolic agents at the 1 μM level do not have a direct anabolic effect on muscle or alter glucocorticoid-induced catabolic response in muscle

  2. Biochemistry of cellulose degradation and cellulose utilization for feeds and for protein

    Energy Technology Data Exchange (ETDEWEB)

    Sadara, J C; Lachke, A H; Shewale, J G

    1979-01-01

    A review discussing production of single-cell protein, fuel, and glucose from cellulose decomposition; surface or solid fermentations of single-cell protein; production of cellulases; and the biochemistry of cellulose degradation was presented.

  3. The different roles of selective autophagic protein degradation in mammalian cells.

    Science.gov (United States)

    Wang, Da-wei; Peng, Zhen-ju; Ren, Guang-fang; Wang, Guang-xin

    2015-11-10

    Autophagy is an intracellular pathway for bulk protein degradation and the removal of damaged organelles by lysosomes. Autophagy was previously thought to be unselective; however, studies have increasingly confirmed that autophagy-mediated protein degradation is highly regulated. Abnormal autophagic protein degradation has been associated with multiple human diseases such as cancer, neurological disability and cardiovascular disease; therefore, further elucidation of protein degradation by autophagy may be beneficial for protein-based clinical therapies. Macroautophagy and chaperone-mediated autophagy (CMA) can both participate in selective protein degradation in mammalian cells, but the process is quite different in each case. Here, we summarize the various types of macroautophagy and CMA involved in determining protein degradation. For this summary, we divide the autophagic protein degradation pathways into four categories: the post-translational modification dependent and independent CMA pathways and the ubiquitin dependent and independent macroautophagy pathways, and describe how some non-canonical pathways and modifications such as phosphorylation, acetylation and arginylation can influence protein degradation by the autophagy lysosome system (ALS). Finally, we comment on why autophagy can serve as either diagnostics or therapeutic targets in different human diseases.

  4. Supply of N compounds to the rumen and their subsequent metabolism and nutritional value

    International Nuclear Information System (INIS)

    Smith, R.H.; Grantley-Smith, M.P.; Merry, R.J.; McAllan, A.B.; Oldham, J.D.; Salter, D.N.

    1987-01-01

    Three projects studying different aspects of the utilization of N compounds by young steer are described. (1) Compartmental analysis of 15 N movements between rumen, blood and urine showed that the level of feeding had no effect on the proportion of N intake recycled to the alimentary tract as urea, but a substantial effect on the site of re-entry; at a high level of feeding most went to the rumen, while at a low level it went to the rest of the tract. Some effects of manipulating growth rate with an anabolic agent on N transfers are also described. (2) Digestibilities of fibre and the efficiencies of microbial protein synthesis in the rumens of animals receiving diets containing alkali treated straw were investigated. The former, but not the latter, were significantly improved when dietary rumen degradable nitrogen was supplied as certain true protein sources rather than mainly as urea. This indication of a requirement for exogenous amino acids and/or peptides was supported by in vitro studies using a solid fed continuous culture system. (3) The entry of 3 H and 15 N labels into the portal vein reached a peak within one hour of infusing labelled leucine, glycine, lysine and glutamine into the duodenum. For leucine, glycine and lysine substantial amounts of the labels were newly entered and were still in the free amino acid at this time, but by two hours only recycled label was present, mostly in plasma protein. Similar peaks of label appearance occurred after glutamine infusion, but these were mainly in glutamic acid and arginine and entry from the gut continued for several hours. The patterns of appearance and disappearance of 35 S following duodenal infusion of labelled methionine were similar to those for 3 H following leucine infusion. (author)

  5. Degradation of misfolded proteins in neurodegenerative diseases: therapeutic targets and strategies.

    Science.gov (United States)

    Ciechanover, Aaron; Kwon, Yong Tae

    2015-03-13

    Mammalian cells remove misfolded proteins using various proteolytic systems, including the ubiquitin (Ub)-proteasome system (UPS), chaperone mediated autophagy (CMA) and macroautophagy. The majority of misfolded proteins are degraded by the UPS, in which Ub-conjugated substrates are deubiquitinated, unfolded and cleaved into small peptides when passing through the narrow chamber of the proteasome. The substrates that expose a specific degradation signal, the KFERQ sequence motif, can be delivered to and degraded in lysosomes via the CMA. Aggregation-prone substrates resistant to both the UPS and the CMA can be degraded by macroautophagy, in which cargoes are segregated into autophagosomes before degradation by lysosomal hydrolases. Although most misfolded and aggregated proteins in the human proteome can be degraded by cellular protein quality control, some native and mutant proteins prone to aggregation into β-sheet-enriched oligomers are resistant to all known proteolytic pathways and can thus grow into inclusion bodies or extracellular plaques. The accumulation of protease-resistant misfolded and aggregated proteins is a common mechanism underlying protein misfolding disorders, including neurodegenerative diseases such as Huntington's disease (HD), Alzheimer's disease (AD), Parkinson's disease (PD), prion diseases and Amyotrophic Lateral Sclerosis (ALS). In this review, we provide an overview of the proteolytic pathways in neurons, with an emphasis on the UPS, CMA and macroautophagy, and discuss the role of protein quality control in the degradation of pathogenic proteins in neurodegenerative diseases. Additionally, we examine existing putative therapeutic strategies to efficiently remove cytotoxic proteins from degenerating neurons.

  6. A genetic bistable switch utilizing nonlinear protein degradation.

    Science.gov (United States)

    Huang, Daniel; Holtz, William J; Maharbiz, Michel M

    2012-07-09

    Bistability is a fundamental property in engineered and natural systems, conferring the ability to switch and retain states. Synthetic bistable switches in prokaryotes have mainly utilized transcriptional components in their construction. Using both transcriptional and enzymatic components, creating a hybrid system, allows for wider bistable parameter ranges in a circuit. In this paper, we demonstrate a tunable family of hybrid bistable switches in E. coli using both transcriptional components and an enzymatic component. The design contains two linked positive feedback loops. The first loop utilizes the lambda repressor, CI, and the second positive feedback loop incorporates the Lon protease found in Mesoplasma florum (mf-Lon). We experimentally tested for bistable behavior in exponential growth phase, and found that our hybrid bistable switch was able to retain its state in the absence of an input signal throughout 40 cycles of cell division. We also tested the transient behavior of our switch and found that switching speeds can be tuned by changing the expression rate of mf-Lon. To our knowledge, this work demonstrates the first use of dynamic expression of an orthogonal and heterologous protease to tune a nonlinear protein degradation circuit. The hybrid switch is potentially a more robust and tunable topology for use in prokaryotic systems.

  7. Effects of gamma irradiation on chemical composition and ruminal protein degradation of canola meal

    Science.gov (United States)

    Shawrang, P.; Nikkhah, A.; Zare-Shahneh, A.; Sadeghi, A. A.; Raisali, G.; Moradi-Shahrebabak, M.

    2008-07-01

    Gamma irradiation of canola meal (at doses of 25, 50 and 75 kGy) could alter its ruminal protein degradation characteristics by cross-linking of the polypeptide chains. This processing resulted in decrease (linear effect, Pruminal protein degradation and increase (linear effect, Pruminant nutrition.

  8. Comparison of rumen bacteria distribution in original rumen digesta, rumen liquid and solid fractions in lactating Holstein cows

    OpenAIRE

    Ji, Shoukun; Zhang, Hongtao; Yan, Hui; Azarfar, Arash; Shi, Haitao; Alugongo, Gibson; Li, Shengli; Cao, Zhijun; Wang, Yajing

    2017-01-01

    Background Original rumen digesta, rumen liquid and solid fractions have been frequently used to assess the rumen bacterial community. However, bacterial profiles in rumen original digesta, liquid and solid fractions vary from each other and need to be better established. Methods To compare bacterial profiles in each fraction, samples of rumen digesta from six cows fed either a high fiber diet (HFD) or a high energy diet (HED) were collected via rumen fistulas. Rumen digesta was then squeezed...

  9. 常用饲料原料蛋白质在梅花鹿瘤胃内降解率的测定%Determination of Protein Ruminal Degradability of Common Feed Ingredients in Sika Deer

    Institute of Scientific and Technical Information of China (English)

    鲍坤; 徐超; 宁浩然; 王凯英; 赵家平; 李光玉

    2012-01-01

    : In order to investigate the protein degradation rule in rumen of several common feed ingredients in deer, four adult male sika deer fitted with permanent rumen cannulas were selected to estimate the protein ruminal degradability of cottonseed meal, corn germ meal, rapeseed meal, distillers' dried grains with solubles (DDGS) , corn gluten meal, corn fiber and Chinese wildrye using the nylon bag technology. The results showed as follows: protein ruminal degradability of cottonseed meal was always the highest, compared with other common feed ingredients, the difference in protein ruminal degradability at all time points was extremely significant (P<0.01). 2) The protein ruminal degradability at 48 hour in descending order was cottonseed meal, corn gluten meal, Chinese wildrye, DDGS, corn germ meal, rapeseed meal and corn fiber, and the protein ruminal dynamic degradability showed the similar change law. It is concluded that protection technology of protein bypass rumen must be used in practice to reduce waste of protein source because of high degradability of cottonseed meal; rapeseed meal is a kind of new protein supplement to develop because of its low protein ruminal degradability; corn germ meal, DDGS, corn gluten meal, corn fiber and Chinese wildrye can be used as common feed ingredients in deer production. [ Chinese Journal of Animal Nutrition, 2012, 24 (11) : 2257-2262]%为研究几种鹿常用饲料原料的蛋白质瘤胃降解规律,以4头安装有永久性瘤胃瘘管的成年雄性梅花鹿为试验动物,采用尼龙袋法对棉籽粕、玉米胚芽粕、菜籽粕、干酒糟及其可溶物(DDGS)、玉米蛋白粉、玉米纤维及羊草的蛋白质瘤胃降解率进行测定.结果表明:1)棉籽粕的蛋白质瘤胃降解率始终最高,与其他几种饲料原料相比,在各时间点的差异均达到极显著(P<0.01).2)48 h的蛋白质瘤胃降解率从高到低依次为棉籽粕、玉米蛋白粉、羊草、DDGS、玉米胚芽粕、菜籽粕

  10. Chemical degradation of proteins in the solid state with a focus on photochemical reactions.

    Science.gov (United States)

    Mozziconacci, Olivier; Schöneich, Christian

    2015-10-01

    Protein pharmaceuticals comprise an increasing fraction of marketed products but the limited solution stability of proteins requires considerable research effort to prepare stable formulations. An alternative is solid formulation, as proteins in the solid state are thermodynamically less susceptible to degradation. Nevertheless, within the time of storage a large panel of kinetically controlled degradation reactions can occur such as, e.g., hydrolysis reactions, the formation of diketopiperazine, condensation and aggregation reactions. These mechanisms of degradation in protein solids are relatively well covered by the literature. Considerably less is known about oxidative and photochemical reactions of solid proteins. This review will provide an overview over photolytic and non-photolytic degradation reactions, and specially emphasize mechanistic details on how solid structure may affect the interaction of protein solids with light. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Effects of Temperature during Moist Heat Treatment on Ruminal Degradability and Intestinal Digestibility of Protein and Amino Acids in Hempseed Cake.

    Science.gov (United States)

    Karlsson, L; Ruiz-Moreno, M; Stern, M D; Martinsson, K

    2012-11-01

    The objective of this study was to evaluate ruminal degradability and intestinal digestibility of crude protein (CP) and amino acids (AA) in hempseed cake (HC) that were moist heat treated at different temperatures. Samples of cold-pressed HC were autoclaved for 30 min at 110, 120 or 130°C, and a sample of untreated HC was used as the control. Ruminal degradability of CP was estimated, using the in situ Dacron bag technique; intestinal CP digestibility was estimated for the 16 h in situ residue using a three-step in vitro procedure. AA content was determined for the HC samples (heat treated and untreated) of the intact feed, the 16 h in situ residue and the residue after the three-step procedure. There was a linear increase in RUP (p = 0.001) and intestinal digestibility of RUP (p = 0.003) with increasing temperature during heat treatment. The 130°C treatment increased RUP from 259 to 629 g/kg CP, while intestinal digestibility increased from 176 to 730 g/kg RUP, compared to the control. Hence, the intestinal available dietary CP increased more than eight times. Increasing temperatures during heat treatment resulted in linear decreases in ruminal degradability of total AA (p = 0.006) and individual AA (pruminal degradability of total AA from 837 to 471 g/kg, while intestinal digestibility increased from 267 to 813 g/kg of rumen undegradable AA, compared with the control. There were differences between ruminal AA degradability and between intestinal AA digestibility within all individual HC treatments (pruminants.

  12. Prefoldin Promotes Proteasomal Degradation of Cytosolic Proteins with Missense Mutations by Maintaining Substrate Solubility.

    Directory of Open Access Journals (Sweden)

    Sophie A Comyn

    2016-07-01

    Full Text Available Misfolded proteins challenge the ability of cells to maintain protein homeostasis and can accumulate into toxic protein aggregates. As a consequence, cells have adopted a number of protein quality control pathways to prevent protein aggregation, promote protein folding, and target terminally misfolded proteins for degradation. In this study, we employed a thermosensitive allele of the yeast Guk1 guanylate kinase as a model misfolded protein to investigate degradative protein quality control pathways. We performed a flow cytometry based screen to identify factors that promote proteasomal degradation of proteins misfolded as the result of missense mutations. In addition to the E3 ubiquitin ligase Ubr1, we identified the prefoldin chaperone subunit Gim3 as an important quality control factor. Whereas the absence of GIM3 did not impair proteasomal function or the ubiquitination of the model substrate, it led to the accumulation of the poorly soluble model substrate in cellular inclusions that was accompanied by delayed degradation. We found that Gim3 interacted with the Guk1 mutant allele and propose that prefoldin promotes the degradation of the unstable model substrate by maintaining the solubility of the misfolded protein. We also demonstrated that in addition to the Guk1 mutant, prefoldin can stabilize other misfolded cytosolic proteins containing missense mutations.

  13. Metabolisable protein supply to lactating dairy cows increased with increasing dry matter concentration in grass-clover silage

    DEFF Research Database (Denmark)

    Johansen, Marianne; Hellwing, Anne Louise Frydendahl; Lund, Peter

    2017-01-01

    The aim of this experiment was to study the effect of increased dry matter (DM) concentration in grass-clover silage, obtained by extending the pre-wilting period before ensiling, on the amount of metabolisable protein (MP) supplied to lactating dairy cows. Spring growth and first regrowth of grass...... and faeces, respectively, were collected over 94 h to cover the diurnal variation, pooled, and subsequently analysed. Rumen fluid was collected in same sampling procedure. To estimate the duodenal flow of microbial protein, microbes were isolated from the rumen and analysed for amino acids (AA) and purines...... flow of AA. The higher duodenal flow of AA derived from a lower rumen degradation of feed protein and a tendency towards a higher microbial synthesis in the rumen. Fibre digestibility and CH4 production were not affected by silage DM concentration. In conclusion, MP concentration in grass-clover silage...

  14. Protein synthesis and degradation during starvation-induced cardiac atrophy in rabbits

    International Nuclear Information System (INIS)

    Samarel, A.M.; Parmacek, M.S.; Magid, N.M.; Decker, R.S.; Lesch, M.

    1987-01-01

    To determine the relative importance of protein degradation in the development of starvation-induced cardiac atrophy, in vivo fractional synthetic rates of total cardiac protein, myosin heavy chain, actin, light chain 1, and light chain 2 were measured in fed and fasted rabbits by continuous infusion of [ 3 H] leucine. In addition, the rate of left ventricular protein accumulation and loss were assessed in weight-matched control and fasted rabbits. Rates of total cardiac protein degradation were then estimated as the difference between rates of synthesis and growth. Fasting produced left ventricular atrophy by decreasing the rate of left ventricular protein synthesis (34.8 +/- 1.4, 27.3 +/- 3.0, and 19.3 +/- 1.2 mg/day of left ventricular protein synthesized for 0-, 3-, and 7-day fasted rabbits, respectively). Inhibition of contractile protein synthesis was evident by significant reductions in the fractional synthetic rates of all myofibrillar protein subunits. Although fractional rates of protein degradation increased significantly within 7 days of fasting, actual amounts of left ventricular protein degraded per day were unaffected. Thus, prolonged fasting profoundly inhibits the synthesis of new cardiac protein, including the major protein constituents of the myofibril. Both this inhibition in new protein synthesis as well as a smaller but significant reduction in the average half-lives of cardiac proteins are responsible for atrophy of the heart in response to fasting

  15. In vitro degradation of the 32kDa PS II reaction centre protein

    International Nuclear Information System (INIS)

    Eckenswiller, L.C.; Greenberg, B.M.

    1989-01-01

    The 32kDa thylakoid membrane protein is an integral component of the PS II reaction centre. The protein, although stable in the dark, undergoes light dependent turnover. Light from the UV, visible and far-red spectral regions induce 32kDa protein degradation. To better understand 32kDa protein metabolism, an in vitro degradation system is being developed. It consists of isolated thylakoid membranes than contain radiolabelled protein. The 32kDa protein is actively and specifically degraded when the thylakoid preparation is exposed to UV or visible radiation. The protein is stable in the dark. The herbicides (atrazine and DCMU) inhibit degradation in the in vitro system as they do in vivo. Additionally, several methods of isolating thylakoids are being compared to optimize the 32kDa protein degradation reaction. The preparations will be evaluated based on their ability to permit light dependent degradation of the 32kDa protein without affecting the other membrane components

  16. Sequence and 3D structure based analysis of TNT degrading proteins in Arabidopsis thaliana.

    Science.gov (United States)

    Bhattacherjee, Amrita; Mandal, Rahul Shubhra; Das, Santasabuj; Kundu, Sudip

    2014-03-01

    TNT, accidentally released at several manufacturing sites, contaminates ground water and soil. It has a toxic effect to algae and invertebrate, and chronic exposure to TNT also causes harmful effects to human. On the other hand, many plants including Arabidopsis thaliana have the ability to metabolize TNT either completely or at least to a reduced less toxic form. In A. thaliana, the enzyme UDP glucosyltransferase (UDPGT) can further conjugate the reduced forms 2-HADNT and 4-HADNT (2-hydroxylamino-4, 6- dinitrotoluene and 4-hydroxylamino-2, 6- dinitrotoluene) of TNT. Based on the experimental analysis, existing literature and phylogenetic analysis, it is evident that among 107 UDPGT proteins only six are involved in the TNT degrading process. A total of 13 UDPGT proteins including five of these TNT degrading proteins fall within the same group of phylogeny. Thus, these 13 UDPGT proteins have been classified into two groups, TNT-degrading and TNT-non-degrading proteins. To understand the differences in TNT-degrading capacities; using homology modeling we first predicted two structures, taking one representative sequence from both the groups. Next, we performed molecular docking of the modeled structure and TNT reduced form 2-hydroxylamino-4, 6- dinitrotoluene (2-HADNT). We observed that while the Trp residue located within the active site region of the TNT- degrading protein showed π-Cation interaction; such type of interaction was absent in TNT-non-degrading protein, as the respective Trp residue lay outside of the pocket in this case. We observed the conservation of this π-Cation interaction during MD simulation of TNT-degrading protein. Thus, the position and the orientation of the active site residue Trp could explain the presence and absence of TNT-degrading capacity of the UDPGT proteins.

  17. Magnitude Differences in Bioactive Compounds, Chemical Functional Groups, Fatty Acid Profiles, Nutrient Degradation and Digestion, Molecular Structure, and Metabolic Characteristics of Protein in Newly Developed Yellow-Seeded and Black-Seeded Canola Lines.

    Science.gov (United States)

    Theodoridou, Katerina; Zhang, Xuewei; Vail, Sally; Yu, Peiqiang

    2015-06-10

    Recently, new lines of yellow-seeded (CS-Y) and black-seeded canola (CS-B) have been developed with chemical and structural alteration through modern breeding technology. However, no systematic study was found on the bioactive compounds, chemical functional groups, fatty acid profiles, inherent structure, nutrient degradation and absorption, or metabolic characteristics between the newly developed yellow- and black-seeded canola lines. This study aimed to systematically characterize chemical, structural, and nutritional features in these canola lines. The parameters accessed include bioactive compounds and antinutrition factors, chemical functional groups, detailed chemical and nutrient profiles, energy value, nutrient fractions, protein structure, degradation kinetics, intestinal digestion, true intestinal protein supply, and feed milk value. The results showed that the CS-Y line was lower (P ≤ 0.05) in neutral detergent fiber (122 vs 154 g/kg DM), acid detergent fiber (61 vs 99 g/kg DM), lignin (58 vs 77 g/kg DM), nonprotein nitrogen (56 vs 68 g/kg DM), and acid detergent insoluble protein (11 vs 35 g/kg DM) than the CS-B line. There was no difference in fatty acid profiles except C20:1 eicosenoic acid content (omega-9) which was in lower in the CS-Y line (P structure spectral profile, there were no significant differences in functional groups of amides I and II, α helix, and β-sheet structure as well as their ratio between the two new lines, indicating no difference in protein structure makeup and conformation between the two lines. In terms of energy values, there were significant differences in total digestible nutrient (TDN; 149 vs 133 g/kg DM), metabolizable energy (ME; 58 vs 52 MJ/kg DM), and net energy for lactation (NEL; 42 vs 37 MJ/kg DM) between CS-Y and CS-B lines. For in situ rumen degradation kinetics, the two lines differed in soluble fraction (S; 284 vs 341 g/kg CP), potential degradation fraction (D; 672 vs 590 g/kg CP), and effective degraded

  18. Fontes protéicas e energéticas com diferentes degradabilidades ruminais para novilhos de corte = Protein and Energy sources with differing degradabilities for finishing steers

    Directory of Open Access Journals (Sweden)

    Paola Ranzani Gabarra

    2007-04-01

    Full Text Available O objetivo deste trabalho foi determinar o efeito da sincronização dadegradação ruminal de fontes de amido (milho moído fino ou milho floculado e proteína (farelo de soja e uréia, no consumo de matéria seca, digestibilidade no trato total e parâmetros ruminais de novilhos Nelore em terminação. Quatro novilhos Nelore (300 kg PV foram utilizados em delineamento do tipo Quadrado Latino 4 x 4, fatorial 2 x 2: dois métodos de processamento do milho (moagem fina x floculação e duas fontes protéicas (farelo de soja x uréia. As rações continham 13% de feno de gramínea e 87% de concentrado. A floculação do milho reduziu a concentração de amido duodenal (p The objective of this study was to determine the effect of synchronization of starch (finely ground or steam-flaked corn and protein (soybean meal or urea rumen degradation on dry matter intake, total tract digestibility and rumen parameters of finishingNelore steers. Four Nelore steers (300 kg LW were utilized in a 4 x 4 Latin Square design, 2 x 2 factorial arragment: two corn processing methods (fine grinding vs. steam flaking and two protein sources (soybean meal vs. urea. The diets contained 13% grass hay and 87%concentrate. Steam flaking decreased (p < 0.01 starch concentration in duodenal digesta, which would explain lower ruminal pH (p < 0.15 and concentration of N-NH3 (p < 0.01, plasma urea-N concentration (p < 0.01; and the increased molar concentration of ruminal propionate (p < 0.01. As compared to fine grinding, steam-flaking corn increased(p < 0.01 total tract digestibility of starch (98.8 vs. 88.6%, but decreased (p < 0.01 NDF digestibility (41.9 vs. 12.1%. Protein sources had no effect on the evaluated variables. Increasing starch degradability through steam-flaking of corn improved dietary proteinutilization by beef steers.

  19. Rumen fermentation and nutrient flow to the omasum in Holstein cows fed extruded canola seeds treated with or without lignosulfonate

    Directory of Open Access Journals (Sweden)

    Wallacy Barbacena Rosa dos Santos

    2012-07-01

    Full Text Available Four multiparous Holstein cows averaging 548 kg of body weight and 74 d in lactation were used in a Latin square design with four 21-d experimental periods to determine effects of feeding extruded versus non-extruded canola seed, with or without 50 g/kg lignosulfonate on rumen fermentation, nutrient flow to the omasum, and degradability of dry matter (DM and N of each diet. The DM effective degradability increased with extrusion and lignosulfonate treatment had no effect. The effective degradability of N was similar between diets. Lignosulfonate treatment of extruded versus non-extruded canola seeds decreased ruminal and total tract apparent digestibility of organic matter. The lowest apparent ruminal and highest intestinal digestibilities of protein, expressed as a percentage of N intake were observed for cows fed extruded canola seeds without lignosulfonate. Lignosulfonate treatment and extrusion had no effect on pH and concentrations of ammonia N and volatile fatty acids in the rumen. Results suggest that extruded canola seed untreated with formaldehyde may stimulate efficiency of microbial protein synthesis and is an effective means of increasing the availability of protein in the small intestine without affecting the total tract apparent digestibility of protein.

  20. A Critical Appraisal of Quantitative Studies of Protein Degradation in the Framework of Cellular Proteostasis

    Directory of Open Access Journals (Sweden)

    Beatriz Alvarez-Castelao

    2012-01-01

    Full Text Available Protein homeostasis, proteostasis, is essential to understand cell function. Protein degradation is a crucial component of the proteostatic mechanisms of the cell. Experiments on protein degradation are nowadays present in many investigations in the field of molecular and cell biology. In the present paper, we focus on the different experimental approaches to study protein degradation and present a critical appraisal of the results derived from steady-state and kinetic experiments using detection of unlabelled and labelled protein methodologies with a proteostatic perspective. This perspective allows pinpointing the limitations in interpretation of results and the need of further experiments and/or controls to establish “definitive evidence” for the role of protein degradation in the proteostasis of a given protein or the entire proteome. We also provide a spreadsheet for simple calculations of mRNA and protein decays for mimicking different experimental conditions and a checklist for the analysis of experiments dealing with protein degradation studies that may be useful for researchers interested in the area of protein turnover.

  1. Degradation of protein translation machinery by amino acid starvation-induced macroautophagy

    DEFF Research Database (Denmark)

    Gretzmeier, Christine; Eiselein, Sven; Johnson, Gregory R.

    2017-01-01

    , unbiased approaches relying on quantitative mass spectrometry-based proteomics. Macroautophagy is induced by rapamycin treatment, and by amino acid and glucose starvation in differentially, metabolically labeled cells. Protein dynamics are linked to image-based models of autophagosome turnover. Depending...... on the inducing stimulus, protein as well as organelle turnover differ. Amino acid starvation-induced macroautophagy leads to selective degradation of proteins important for protein translation. Thus, protein dynamics reflect cellular conditions in the respective treatment indicating stimulus-specific pathways...

  2. Toscana virus NSs protein promotes degradation of double-stranded RNA-dependent protein kinase.

    Science.gov (United States)

    Kalveram, Birte; Ikegami, Tetsuro

    2013-04-01

    Toscana virus (TOSV), which is transmitted by Phlebotomus spp. sandflies, is a major etiologic agent of aseptic meningitis and encephalitis in the Mediterranean. Like other members of the genus Phlebovirus of the family Bunyaviridae, TOSV encodes a nonstructural protein (NSs) in its small RNA segment. Although the NSs of Rift Valley fever virus (RVFV) has been identified as an important virulence factor, which suppresses host general transcription, inhibits transcription from the beta interferon promoter, and promotes the proteasomal degradation of double-stranded RNA-dependent protein kinase (PKR), little is known about the functions of NSs proteins encoded by less-pathogenic members of this genus. In this study we report that TOSV is able to downregulate PKR with similar efficiency as RVFV, while infection with the other phleboviruses-i.e., Punta Toro virus, sandfly fever Sicilian virus, or Frijoles virus-has no effect on cellular PKR levels. In contrast to RVFV, however, cellular transcription remains unaffected during TOSV infection. TOSV NSs protein promotes the proteasome-dependent downregulation of PKR and is able to interact with kinase-inactive PKR in infected cells.

  3. The Arabidopsis CROWDED NUCLEI genes regulate seed germination by modulating degradation of ABI5 protein.

    Science.gov (United States)

    Zhao, Wenming; Guan, Chunmei; Feng, Jian; Liang, Yan; Zhan, Ni; Zuo, Jianru; Ren, Bo

    2016-07-01

    In Arabidopsis, the phytohormone abscisic acid (ABA) plays a vital role in inhibiting seed germination and in post-germination seedling establishment. In the ABA signaling pathway, ABI5, a basic Leu zipper transcription factor, has important functions in the regulation of seed germination. ABI5 protein localizes in nuclear bodies, along with AFP, COP1, and SIZ1, and was degraded through the 26S proteasome pathway. However, the mechanisms of ABI5 nuclear body formation and ABI5 protein degradation remain obscure. In this study, we found that the Arabidopsis CROWDED NUCLEI (CRWN) proteins, predicted nuclear matrix proteins essential for maintenance of nuclear morphology, also participate in ABA-controlled seed germination by regulating the degradation of ABI5 protein. During seed germination, the crwn mutants are hypersensitive to ABA and have higher levels of ABI5 protein compared to wild type. Genetic analysis suggested that CRWNs act upstream of ABI5. The observation that CRWN3 colocalizes with ABI5 in nuclear bodies indicates that CRWNs might participate in ABI5 protein degradation in nuclear bodies. Moreover, we revealed that the extreme C-terminal of CRWN3 protein is necessary for its function in the response to ABA in germination. Our results suggested important roles of CRWNs in ABI5 nuclear body organization and ABI5 protein degradation during seed germination. © 2015 Institute of Botany, Chinese Academy of Sciences.

  4. Muscle protein degradation and amino acid metabolism during prolonged knee-extensor exercise in humans

    DEFF Research Database (Denmark)

    Van Hall, Gerrit; Saltin, B; Wagenmakers, A J

    1999-01-01

    to a substantial increase in net muscle protein degradation, and that a lowering of the starting muscle glycogen content leads to a further increase. The carbon atoms of the branched-chain amino acids (BCAA), glutamate, aspartate and asparagine, liberated by protein degradation, and the BCAA and glutamate......The aim of this study was to investigate whether prolonged one-leg knee-extensor exercise enhances net protein degradation in muscle with a normal or low glycogen content. Net amino acid production, as a measure of net protein degradation, was estimated from leg exchange and from changes...... in the concentrations of amino acids that are not metabolized in skeletal muscle. Experiments were performed at rest and during one-leg knee-extensor exercise in six subjects having one leg with a normal glycogen content and the other with a low glycogen content. Exercise was performed for 90 min at a workload of 60...

  5. Highly Promiscuous Oxidases Discovered in the Bovine Rumen Microbiome

    Directory of Open Access Journals (Sweden)

    Lisa Ufarté

    2018-05-01

    Full Text Available The bovine rumen hosts a diverse microbiota, which is highly specialized in the degradation of lignocellulose. Ruminal bacteria, in particular, are well equipped to deconstruct plant cell wall polysaccharides. Nevertheless, their potential role in the breakdown of the lignin network has never been investigated. In this study, we used functional metagenomics to identify bacterial redox enzymes acting on polyaromatic compounds. A new methodology was developed to explore the potential of uncultured microbes to degrade lignin derivatives, namely kraft lignin and lignosulfonate. From a fosmid library covering 0.7 Gb of metagenomic DNA, three hit clones were identified, producing enzymes able to oxidize a wide variety of polyaromatic compounds without the need for the addition of copper, manganese, or mediators. These promiscuous redox enzymes could thus be of potential interest both in plant biomass refining and dye remediation. The enzymes were derived from uncultured Clostridia, and belong to complex gene clusters involving proteins of different functional types, including hemicellulases, which likely work in synergy to produce substrate degradation.

  6. Degradation of extracytoplasmic catalysts for protein folding in Bacillus subtilis

    NARCIS (Netherlands)

    Krishnappa, Laxmi; Monteferrante, Carmine G; Neef, Jolanda; Dreisbach, Annette; van Dijl, Jan Maarten

    The general protein secretion pathway of Bacillus subtilis has a high capacity for protein export from the cytoplasm, which is exploited in the biotechnological production of a wide range of enzymes. These exported proteins pass the membrane in an unfolded state, and accordingly, they have to fold

  7. Ruminal Degradability of Dry Matter and Crude Protein from Moist Dehulled Lupin and Extruded Rapeseed Meal Degradabilidad Ruminal de la Materia Seca y de la Proteína Cruda de Lupino Descascarado y Torta de Raps Extruidos

    Directory of Open Access Journals (Sweden)

    Claudia Barchiesi-Ferrari

    2011-09-01

    Full Text Available The flow of ruminal undegradable protein (RUP to the small intestine can be increased if ruminal degradation of dietary protein is reduced. The objective of this study was to evaluate the effect of extrusion on ruminal degradability of dry matter (DM and crude protein (CP from dehulled lupin (Lupinus albus L. (DL and rapeseed (Brassica napus L. meal (RM. Unextruded soybean (Glicine max L. meal (SBM was used as a control. The DL was extruded at 130 ºC with 20% moisture and RM was extruded at 120 ºC with 20% moisture. Ruminal degradability was evaluated in situ by incubating feed samples for 2, 4, 8, 12, 24, and 48 h of fermentation in the rumen using three rumen-fistulated dairy cows. Values of CP soluble fraction (“a” in SBM, DL, extruded dehulled lupin (EDL, RM, and extruded rapeseed meal (ERM was lower in the extruded feeds (P El flujo de proteína no degradable en el rumen (RUP hacia el intestino delgado puede ser incrementado si se reduce la degradación ruminal de la proteína dietaria. El objetivo de este trabajo fue evaluar el efecto de la extrusión sobre la degradabilidad ruminal de la materia seca (DM y proteína cruda (CP de lupino (Lupinus albus L. descascarado (DL y torta de raps (Brassica napus L. (RM. Se utilizó afrecho de soya (Glicine max L. sin extruir (SBM como control. El DL fue extruido a 130 ºC con 20% de humedad y la RM fue extruida a 120 ºC con 20% de humedad. La degradabilidad ruminal se evaluó in situ incubando las muestras de alimentos a 2, 4, 8, 12, 24 y 48 h de fermentación en tres vacas lecheras con fístula ruminal. Los valores de la fracción soluble de la CP (“a” en SBM, en DL, lupino descascarado extruido (EDL, RM y torta de raps extruida (ERM fue menor en los extruidos (P < 0.05. La fracción lentamente degradable (“b” de SBM, DL, EDL, RM y de ERM fue 858; 593; 622 y 451 y 457 g kg-1, respectivamente, y se incrementó por extrusión (P < 0.05. La extrusión redujo la degradabilidad efectiva

  8. Effects of gamma irradiation on chemical composition and ruminal protein degradation of canola meal

    Energy Technology Data Exchange (ETDEWEB)

    Shawrang, P. [Agriculture, Medical and Industrial Research School, Nuclear Science and Technology Research Institute, Atomic Energy Organization of Iran, P.O. Box 31485-498, Karaj (Iran, Islamic Republic of); Department of Animal Science, Faculty of Agriculture, Tehran University P.O. Box 4111, Karaj (Iran, Islamic Republic of)], E-mail: parvinshawrang@yahoo.co.uk; Nikkhah, A.; Zare-Shahneh, A. [Department of Animal Science, Faculty of Agriculture, Tehran University P.O. Box 4111, Karaj (Iran, Islamic Republic of); Sadeghi, A.A. [Department of Animal Science, Faculty of Agriculture, Science and Research Branch, Islamic Azad University, P.O. Box 14515-4933, Tehran (Iran, Islamic Republic of); Raisali, G. [Radiation Applications Research School, Nuclear Science and Technology Research Institute, Atomic Energy Organization of Iran, P.O. Box 11365-3486, Tehran (Iran, Islamic Republic of); Moradi-Shahrebabak, M. [Department of Animal Science, Faculty of Agriculture, Tehran University P.O. Box 4111, Karaj (Iran, Islamic Republic of)

    2008-07-15

    Gamma irradiation of canola meal (at doses of 25, 50 and 75 kGy) could alter its ruminal protein degradation characteristics by cross-linking of the polypeptide chains. This processing resulted in decrease (linear effect, P<0.001) of ruminal protein degradation and increase (linear effect, P<0.001) of intestinal protein digestibility. The results showed that gamma irradiation at doses higher than 25 kGy can be used as a cross-linking agent to improve protein properties of supplements in ruminant nutrition.

  9. Effects of gamma irradiation on chemical composition and ruminal protein degradation of canola meal

    International Nuclear Information System (INIS)

    Shawrang, P.; Nikkhah, A.; Zare-Shahneh, A.; Sadeghi, A.A.; Raisali, G.; Moradi-Shahrebabak, M.

    2008-01-01

    Gamma irradiation of canola meal (at doses of 25, 50 and 75 kGy) could alter its ruminal protein degradation characteristics by cross-linking of the polypeptide chains. This processing resulted in decrease (linear effect, P<0.001) of ruminal protein degradation and increase (linear effect, P<0.001) of intestinal protein digestibility. The results showed that gamma irradiation at doses higher than 25 kGy can be used as a cross-linking agent to improve protein properties of supplements in ruminant nutrition

  10. Effects of replacing soybean meal with canola meal differing in rumen-undegradable protein content on ruminal fermentation and gas production kinetics using 2 in vitro systems.

    Science.gov (United States)

    Paula, E M; Monteiro, H F; Silva, L G; Benedeti, P D B; Daniel, J L P; Shenkoru, T; Broderick, G A; Faciola, A P

    2017-07-01

    Previous research indicated that there were significant differences in rumen-undegradable protein (RUP) among canola meals (CM), which could influence the nutritional value of CM. The objectives of this study were to (1) evaluate the effects of feeding CM with different RUP contents on ruminal fermentation, nutrient digestion, and microbial growth using a dual-flow continuous culture system (experiment 1) and (2) evaluate ruminal gas production kinetics, in vitro organic matter (OM) digestibility, and methane (CH 4 ) production of soybean meal (SBM) and CM with low or high RUP in the diet or as a sole ingredient using a gas production system (experiments 2 and 3). In experiment 1, diets were randomly assigned to 6 fermentors in a replicated 3 × 3 Latin square. The only ingredient that differed among diets was the protein supplement. The treatments were (1) solvent-extracted SBM, (2) low-RUP solvent-extracted CM (38% RUP as a percentage of crude protein), and (3) high-RUP solvent-extracted CM (50% RUP). Diets were prepared as 3 concentrate mixtures that were combined with 25% orchardgrass hay and 15% wheat straw (dry matter basis). Experiments 2 and 3 had the same design with 24 bottles incubated 3 times for 48 h each. During the 48-h incubation, the cumulative pressure was recorded to determine gas production kinetics, in vitro OM digestibility, and CH 4 production. In experiment 1, N flow (g/d), efficiency of N use, efficiency of bacterial N synthesis, total volatile fatty acids (mM), and molar proportion of acetate, propionate, and isobutyrate were not affected by treatments. There were tendencies for a decrease in ruminal NH 3 -N and an increase in molar proportion of butyrate for the SBM diet compared with both CM diets. The molar proportion of valerate was greater in both CM diets, whereas the molar proportion of isovalerate and total branched-chain volatile fatty acids was lower for the CM diets compared with the SBM diet. In experiments 2 and 3, the SBM

  11. Critical lysine residues of Klf4 required for protein stabilization and degradation

    Energy Technology Data Exchange (ETDEWEB)

    Lim, Key-Hwan; Kim, So-Ra; Ramakrishna, Suresh; Baek, Kwang-Hyun, E-mail: baek@cha.ac.kr

    2014-01-24

    Highlights: • Klf4 undergoes the 26S proteasomal degradation by ubiquitination on its multiple lysine residues. • Essential Klf4 ubiquitination sites are accumulated between 190–263 amino acids. • A mutation of lysine at 232 on Klf4 elongates protein turnover. • Klf4 mutants dramatically suppress p53 expression both under normal and UV irradiated conditions. - Abstract: The transcription factor, Krüppel-like factor 4 (Klf4) plays a crucial role in generating induced pluripotent stem cells (iPSCs). As the ubiquitination and degradation of the Klf4 protein have been suggested to play an important role in its function, the identification of specific lysine sites that are responsible for protein degradation is of prime interest to improve protein stability and function. However, the molecular mechanism regulating proteasomal degradation of the Klf4 is poorly understood. In this study, both the analysis of Klf4 ubiquitination sites using several Klf4 deletion fragments and bioinformatics predictions showed that the lysine sites which are signaling for Klf4 protein degradation lie in its N-terminal domain (aa 1–296). The results also showed that Lys32, 52, 232, and 252 of Klf4 are responsible for the proteolysis of the Klf4 protein. These results suggest that Klf4 undergoes proteasomal degradation and that these lysine residues are critical for Klf4 ubiquitination.

  12. Degradability of dry matter and crude protein of dry grains and wet grain silages from different processing corn hybrids (Zea mays

    Directory of Open Access Journals (Sweden)

    Wagner dos Reis

    2013-03-01

    Full Text Available The objective of this work was to evaluate the effect of processing two corn hybrids conserved, dry and humid grains, the dry matter (DM and crude protein (CP degradability in situ. The particle size was determined and difference was verified in MGD (Medium Geometric Diameter of processed ingredients. Three sheep were used with rumen canulated, in a completely randomized design, using a factorial outline 2 x 2 x 3, being two corn hybrid, two conservation methods and three processing forms (whole, coarsely and finely ground, with five times of incubation (3, 6, 12, 24 and 48 hours. The fraction A in SDC (silage of dent corn of DM was superior to GDC (grain of dent corn in all of the particles size. The ensiling process increased the DM solubility, reducing the fraction B in comparison to dry grain. The values regarding the fractions DP and DE the 5% per hour of the protein, were larger for SDC and GDC, it presents a decreasing when the incubation time advances. The fermentation rate was superior for SDC and GDC. The ensiling process has positive effect in the decreasing of DM and CP in comparison to GDC.

  13. Degradation of structurally characterized proteins injected into HeLa cells. Basic measurements

    International Nuclear Information System (INIS)

    Rogers, S.W.; Rechsteiner, M.

    1988-01-01

    Thirty-five proteins of known x-ray structure were labeled by chloramine-T radioiodination or by reaction with 125I-Bolton-Hunter reagent and introduced into HeLa cells using red cell-mediated microinjection. Degradation rates of the injected proteins were then determined over the next 50 h by measuring the release of soluble isotope to the culture medium. Control experiments demonstrated that the measured rates were not compromised by proteolysis within RBCs, the presence of unfused RBCs, or degradation of protein released from RBCs to the medium. Degradation of some injected proteins was faster during the first 12 h after fusion than at later times, apparently a response of HeLa cells to trypsinization. However, all proteins exhibited first-order degradation rates between 24 and 48 h post injection. Except for seven proteins, stabilities measured during this interval were unaffected by the labeling procedure. Reductive methylation was used to choose among the seven discordant values, and half-lives for the 35 proteins ranged from 16 h for lysozyme to 214 h for yeast alcohol dehydrogenase. Since half-lives for six of the injected proteins closely match values obtained by in vivo measurements, we consider our estimates of the metabolic stabilities of the injected proteins to be generally accurate. Therefore, the half-lives obtained by microinjection should prove useful in the search for relationships between protein structure and intracellular stability

  14. Polyubiquitin chain assembly and organisation determine the dynamics of protein activation and degradation

    Directory of Open Access Journals (Sweden)

    Lan K. Nguyen

    2014-01-01

    Full Text Available Protein degradation via ubiquitination is a major proteolytic mechanism in cells. Once a protein is destined for degradation, it is tagged by multiple ubiquitin molecules. The synthesised polyubiquitin chains can be recognised by the 26S proteosome where proteins are degraded. These chains form through multiple ubiquitination cycles that are similar to multi-site phosphorylation cycles. As kinases and phosphatases, two opposing enzymes (E3 ligases and deubiquitinases DUBs catalyse (deubiquitination cycles. Although multi-ubiquitination cycles are fundamental mechanisms of controlling protein concentrations within a cell, their dynamics have never been explored. Here, we fill this knowledge gap. We show that under permissive physiological conditions, the formation of polyubiquitin chain of length greater than two and subsequent degradation of the ubiquitinated protein, which is balanced by protein synthesis, can display bistable, switch-like responses. Interestingly, the occurrence of bistability becomes pronounced, as the chain grows, giving rise to all-or-none regulation at the protein levels. We give predictions of protein distributions under bistable regime awaiting experimental verification. Importantly, we show for the first time that sustained oscillations can robustly arise in the process of formation of ubiquitin chain, largely due to the degradation of the target protein. This new feature is opposite to the properties of multi-site phosphorylation cycles, which are incapable of generating oscillation if the total abundance of interconverted protein forms is conserved. We derive structural and kinetic constraints for the emergence of oscillations, indicating that a competition between different substrate forms and the E3 and DUB is critical for oscillation. Our work provides the first detailed elucidation of the dynamical features brought about by different molecular setups of the polyubiquitin chain assembly process responsible for

  15. Comparison of Rumen and Manure Microbiomes and Implications for the Inoculation of Anaerobic Digesters.

    Science.gov (United States)

    Ozbayram, Emine Gozde; Ince, Orhan; Ince, Bahar; Harms, Hauke; Kleinsteuber, Sabine

    2018-02-14

    Cattle manure is frequently used as an inoculum for the start-up of agricultural biogas plants or as a co-substrate in the anaerobic digestion of lignocellulosic feedstock. Ruminal microbiota are considered to be effective plant fiber degraders, but the microbes contained in manure do not necessarily reflect the rumen microbiome. The aim of this study was to compare the microbial community composition of cow rumen and manure with respect to plant fiber-digesting microbes. Bacterial and methanogenic communities of rumen and manure samples were examined by 454 amplicon sequencing of bacterial 16S rRNA genes and mcrA genes, respectively. Rumen fluid samples were dominated by Prevotellaceae (29%), whereas Ruminococcaceae was the most abundant family in the manure samples (31%). Fibrobacteraceae (12%) and Bacteroidaceae (13%) were the second most abundant families in rumen fluid and manure, respectively. The high abundances of fiber-degrading bacteria belonging to Prevotellaceae and Fibrobacteraceae might explain the better performance of anaerobic digesters inoculated with rumen fluid. Members of the genus Methanobrevibacter were the predominant methanogens in the rumen fluid, whereas methanogenic communities of the manure samples were dominated by the candidate genus Methanoplasma . Our results suggest that inoculation or bioaugmentation with fiber-digesting rumen microbiota can enhance the anaerobic digestion of lignocellulosic biomass.

  16. Comparison of Rumen and Manure Microbiomes and Implications for the Inoculation of Anaerobic Digesters

    Directory of Open Access Journals (Sweden)

    Emine Gozde Ozbayram

    2018-02-01

    Full Text Available Cattle manure is frequently used as an inoculum for the start-up of agricultural biogas plants or as a co-substrate in the anaerobic digestion of lignocellulosic feedstock. Ruminal microbiota are considered to be effective plant fiber degraders, but the microbes contained in manure do not necessarily reflect the rumen microbiome. The aim of this study was to compare the microbial community composition of cow rumen and manure with respect to plant fiber-digesting microbes. Bacterial and methanogenic communities of rumen and manure samples were examined by 454 amplicon sequencing of bacterial 16S rRNA genes and mcrA genes, respectively. Rumen fluid samples were dominated by Prevotellaceae (29%, whereas Ruminococcaceae was the most abundant family in the manure samples (31%. Fibrobacteraceae (12% and Bacteroidaceae (13% were the second most abundant families in rumen fluid and manure, respectively. The high abundances of fiber-degrading bacteria belonging to Prevotellaceae and Fibrobacteraceae might explain the better performance of anaerobic digesters inoculated with rumen fluid. Members of the genus Methanobrevibacter were the predominant methanogens in the rumen fluid, whereas methanogenic communities of the manure samples were dominated by the candidate genus Methanoplasma. Our results suggest that inoculation or bioaugmentation with fiber-digesting rumen microbiota can enhance the anaerobic digestion of lignocellulosic biomass.

  17. concentrate ratio and rumen ammonia concentration on in situ

    African Journals Online (AJOL)

    The aim of this experiment was to distinguish between the effects of dietary roughage: concentrate ratio and rumen ammonia ... several factors which include the potential rate and extent of degradation of the test feedstuff and the prevailing rate of ... diet effects have be,en designed to examine either the influence of dietary ...

  18. A Structural and Functional Elucidation of the Rumen Microbiome Influenced by Various Diets and Microenvironments

    Directory of Open Access Journals (Sweden)

    Simon Deusch

    2017-08-01

    Full Text Available The structure and function of the microbiome inhabiting the rumen are, amongst other factors, mainly shaped by the animal's feed intake. Describing the influence of different diets on the inherent community arrangement and associated metabolic activities of the most active ruminal fractions (bacteria and archaea is of great interest for animal nutrition, biotechnology, and climatology. Samples were obtained from three fistulated Jersey cows rotationally fed with corn silage, grass silage or grass hay, each supplemented with a concentrate mixture. Samples were fractionated into ruminal fluid, particle-associated rumen liquid, and solid matter. DNA, proteins and metabolites were analyzed subsequently. DNA extracts were used for Illumina sequencing of the 16S rRNA gene and the metabolomes of rumen fluids were determined by 500 MHz-NMR spectroscopy. Tryptic peptides derived from protein extracts were measured by LC-ESI-MS/MS and spectra were processed by a two-step database search for quantitative metaproteome characterization. Data are available via ProteomeXchange with the identifier PXD006070. Protein- and DNA-based datasets revealed significant differences between sample fractions and diets and affirmed similar trends concerning shifts in phylogenetic composition. Ribosomal genes and proteins belonging to the phylum of Proteobacteria, particularly Succinivibrionaceae, exhibited a higher abundance in corn silage-based samples while fiber-degraders of the Lachnospiraceae family emerged in great quantities throughout the solid phase fractions. The analysis of 8163 quantified bacterial proteins revealed the presence of 166 carbohydrate active enzymes in varying abundance. Cellulosome affiliated proteins were less expressed in the grass silage, glycoside hydrolases appeared in slightest numbers in the corn silage. Most expressed glycoside hydrolases belonged to families 57 and 2. Enzymes analogous to ABC transporters for amino acids and

  19. Estimation of rumen microbial protein production from purine derivatives in urine. A laboratory manual for the FAO/IAEA co-ordinated research programme on development, standardization and validation of nuclear based technologies for measuring microbial protein supply in ruminant livestock for improving productivity

    International Nuclear Information System (INIS)

    1997-05-01

    This laboratory manual contains the methodologies used in the standardization and validation of the urine purine derivative technique for estimating microbial protein supply to the rumen. It includes descriptions of methods that involve both radioactive and stable isotopes as well as non isotopic techniques such as chemical assays, since it has been recognised that while isotopic trace techniques provide a powerful tool for nutrition research they can not and should not be used in isolation. Refs, figs, tabs

  20. Evaluation of the contribution of recycled urea to the synthesis of the microbial of protein in the rumen using sup 15 N labelled urea

    Energy Technology Data Exchange (ETDEWEB)

    Al-Dehneh, A M [Michigan State Univ. (USA) Dept. of Animal Science

    1990-01-01

    Urea {sup 15}N was continually infused into the juglar vein for 3 days in two duodenally cannulated cows fed diets of 1:2 and 2:1 forage: concentrate. Duodenal digesta samples were taken every 3 hours, and coccygeal blood and milk were sampled twice daily. Urine was collected for 5 days starting 1 day before infusion and total feces for 3 days during infusion. Fecal samples were also taken twice daily during the 5 days of collection. Urinary excretion of {sup 15}N accounted for about 90% of that which existed from the body; whereas, feces and milk each accounted for about 5%. Recovery of {sup 15}N during the infusion period ranged from 30 to 50% of that infused. Estimates using {sup 15}N ratios, as percent of the total N passing into the duodenum, that was bacterial N, were 50% to 90% and appeared directly proportional to dry matter intake of cows. Recycled-N incorporated into rumen microbes was greater (24% verses 14% of N in bacteria passing into the duodenum) in cows fed the high concentrate than the high forage diet. Also, incorporation of recycled N into rumen microbes was higher in the lactating than the dry cow (24% verses 14%) and the flow of nitrogen from the rumen to the small intestine was greater for the concentrate than the forage diet (122.O% verses 101.0% of nitrogen intake). (author). 27 refs.

  1. Evaluation of the contribution of recycled urea to the synthesis of the microbial of protein in the rumen using 15N labelled urea

    International Nuclear Information System (INIS)

    Al-Dehneh, A.M.

    1990-01-01

    Urea 15 N was continually infused into the juglar vein for 3 days in two duodenally cannulated cows fed diets of 1:2 and 2:1 forage: concentrate. Duodenal digesta samples were taken every 3 hours, and coccygeal blood and milk were sampled twice daily. Urine was collected for 5 days starting 1 day before infusion and total feces for 3 days during infusion. Fecal samples were also taken twice daily during the 5 days of collection. Urinary excretion of 15 N accounted for about 90% of that which existed from the body; whereas, feces and milk each accounted for about 5%. Recovery of 15 N during the infusion period ranged from 30 to 50% of that infused. Estimates using 15 N ratios, as percent of the total N passing into the duodenum, that was bacterial N, were 50% to 90% and appeared directly proportional to dry matter intake of cows. Recycled-N incorporated into rumen microbes was greater (24% verses 14% of N in bacteria passing into the duodenum) in cows fed the high concentrate than the high forage diet. Also, incorporation of recycled N into rumen microbes was higher in the lactating than the dry cow (24% verses 14%) and the flow of nitrogen from the rumen to the small intestine was greater for the concentrate than the forage diet (122.O% verses 101.0% of nitrogen intake). (author). 27 refs

  2. Glucose Deprivation Triggers Protein Kinase C-dependent β-Catenin Proteasomal Degradation*

    Science.gov (United States)

    Choi, Seung-Won; Song, Jun-Kyu; Yim, Ye-Seal; Yun, Ho-Geun; Chun, Kyung-Hee

    2015-01-01

    Autophagy is a conserved process that contributes to cell homeostasis. It is well known that induction mainly occurs in response to nutrient starvation, such as starvation of amino acids and insulin, and its mechanisms have been extensively characterized. However, the mechanisms behind cellular glucose deprivation-induced autophagy are as of now poorly understood. In the present study, we determined a mechanism by which glucose deprivation induced the PKC-dependent proteasomal degradation of β-catenin, leading to autophagy. Glucose deprivation was shown to cause a sub-G1 transition and enhancement of the LC3-II protein levels, whereas β-catenin protein underwent degradation in a proteasome-dependent manner. Moreover, the inhibition of GSK3β was unable to abolish the glucose deprivation-mediated β-catenin degradation or up-regulation of LC3-II protein levels, which suggested GSK3β-independent protein degradation. Intriguingly, the inhibition of PKCα using a pharmacological inhibitor and transfection of siRNA for PKCα was observed to effectively block glucose deprivation-induced β-catenin degradation as well as the increase in LC3-II levels and the accumulation of a sub-G1 population. Together, our results demonstrated a molecular mechanism by which glucose deprivation can induce the GSK3β-independent protein degradation of β-catenin, leading to autophagy. PMID:25691573

  3. Light-induced protein nitration and degradation with HONO emission

    Science.gov (United States)

    Meusel, Hannah; Elshorbany, Yasin; Kuhn, Uwe; Bartels-Rausch, Thorsten; Reinmuth-Selzle, Kathrin; Kampf, Christopher J.; Li, Guo; Wang, Xiaoxiang; Lelieveld, Jos; Pöschl, Ulrich; Hoffmann, Thorsten; Su, Hang; Ammann, Markus; Cheng, Yafang

    2017-10-01

    Proteins can be nitrated by air pollutants (NO2), enhancing their allergenic potential. This work provides insight into protein nitration and subsequent decomposition in the presence of solar radiation. We also investigated light-induced formation of nitrous acid (HONO) from protein surfaces that were nitrated either online with instantaneous gas-phase exposure to NO2 or offline by an efficient nitration agent (tetranitromethane, TNM). Bovine serum albumin (BSA) and ovalbumin (OVA) were used as model substances for proteins. Nitration degrees of about 1 % were derived applying NO2 concentrations of 100 ppb under VIS/UV illuminated conditions, while simultaneous decomposition of (nitrated) proteins was also found during long-term (20 h) irradiation exposure. Measurements of gas exchange on TNM-nitrated proteins revealed that HONO can be formed and released even without contribution of instantaneous heterogeneous NO2 conversion. NO2 exposure was found to increase HONO emissions substantially. In particular, a strong dependence of HONO emissions on light intensity, relative humidity, NO2 concentrations and the applied coating thickness was found. The 20 h long-term studies revealed sustained HONO formation, even when concentrations of the intact (nitrated) proteins were too low to be detected after the gas exchange measurements. A reaction mechanism for the NO2 conversion based on the Langmuir-Hinshelwood kinetics is proposed.

  4. Context Memory Formation Requires Activity-Dependent Protein Degradation in the Hippocampus

    Science.gov (United States)

    Cullen, Patrick K.; Ferrara, Nicole C.; Pullins, Shane E.; Helmstetter, Fred J.

    2017-01-01

    Numerous studies have indicated that the consolidation of contextual fear memories supported by an aversive outcome like footshock requires de novo protein synthesis as well as protein degradation mediated by the ubiquitin-proteasome system (UPS). Context memory formed in the absence of an aversive stimulus by simple exposure to a novel…

  5. Role of Proteasome-Dependent Protein Degradation in Long-Term Operant Memory in "Aplysia"

    Science.gov (United States)

    Lyons, Lisa C.; Gardner, Jacob S.; Gandour, Catherine E.; Krishnan, Harini C.

    2017-01-01

    We investigated the in vivo role of protein degradation during intermediate (ITM) and long-term memory (LTM) in "Aplysia" using an operant learning paradigm. The proteasome inhibitor MG-132 inhibited the induction and molecular consolidation of LTM with no effect on ITM. Remarkably, maintenance of steady-state protein levels through…

  6. Calculating the Degradation Rate of Individual Proteins Using Xenopus Extract Systems.

    Science.gov (United States)

    McDowell, Gary S; Philpott, Anna

    2018-05-16

    The Xenopus extract system has been used extensively as a simple, quick, and robust method for assessing the stability of proteins against proteasomal degradation. In this protocol, methods are provided for assessing the half-life of in vitro translated radiolabeled proteins using Xenopus egg or embryo extracts. © 2019 Cold Spring Harbor Laboratory Press.

  7. Optically degradable dendrons for temporary adhesion of proteins to DNA.

    Science.gov (United States)

    Kostiainen, Mauri A; Kotimaa, Juha; Laukkanen, Marja-Leena; Pavan, Giovanni M

    2010-06-18

    Experimental studies and molecular dynamics modeling demonstrate that multivalent dendrons can be used to temporarily glue proteins and DNA together with high affinity. We describe N-maleimide-cored polyamine dendrons that can be conjugated with free cysteine residues on protein surfaces through 1,4-conjugate addition to give one-to-one protein-polymer conjugates. We used a genetically engineered cysteine mutant of class II hydrophobin (HFBI) and a single-chain Fragment variable (scFv) antibody as model proteins for the conjugation reactions. The binding affinity of the protein-dendron conjugates towards DNA was experimentally assessed by using the ethidium bromide displacement assay. The binding was found to depend on the generation of the dendron, with the second generation having a stronger affinity than the first generation. Thermodynamic parameters of the binding were obtained from molecular dynamics modeling, which showed that the high binding affinity for each system is almost completely driven by a strong favorable binding enthalpy that is opposed by unfavorable binding entropy. A short exposure to UV (lambda approximately 350 nm) can cleave the photolabile o-nitrobenzyl-linked binding ligands from the surface of the dendron, which results in loss of the multivalent binding interactions and triggers the release of the DNA and protein. The timescale of the release is very rapid and the binding partners can be efficiently released after 3 min of UV exposure.

  8. Does altered protein metabolism interfere with postmortem degradation analysis for PMI estimation?

    Science.gov (United States)

    Zissler, A; Ehrenfellner, B; Foditsch, E E; Monticelli, F C; Pittner, S

    2018-03-02

    An accurate estimation of the postmortem interval (PMI) is a central aspect in forensic routine. Recently, a novel approach based on the analysis of postmortem muscle protein degradation has been proposed. However, a number of questions remain to be answered until sensible application of this method to a broad variety of forensic cases is possible. To evaluate whether altered in vivo protein metabolism interferes with postmortem degradation patterns, we conducted a comparative study. We developed a standardized animal degradation model in rats, and collected additional muscle samples from animals recovering from muscle injury and from rats with developed disuse muscle atrophy after induced spinal cord injury. All samples were analyzed by SDS-PAGE and Western blot, labeling well-characterized muscle proteins. Tropomyosin was found to be stable throughout the investigated PMI and no alterations were detected in regenerating and atrophic muscles. In contrast, significant predictable postmortem changes occurred in desmin and vinculin protein band patterns. While no significant deviations from native patterns were detected in at-death samples of disuse muscle atrophy, interestingly, samples of rats recovering from muscle injury revealed additional desmin and vinculin degradation bands that did not occur in this form in any of the examined postmortem samples regardless of PMI. It remains to be investigated whether in vivo-altered metabolism influences postmortem degradation kinetics or if such muscle samples undergo postmortem degradation in a regular fashion.

  9. The mitochondrial translocator protein, TSPO, inhibits HIV-1 envelope glycoprotein biosynthesis via the endoplasmic reticulum-associated protein degradation pathway.

    Science.gov (United States)

    Zhou, Tao; Dang, Ying; Zheng, Yong-Hui

    2014-03-01

    The HIV-1 Env glycoprotein is folded in the endoplasmic reticulum (ER), which is necessary for viral entry and replication. Currently, it is still unclear how this process is regulated. The glycoprotein folding in the ER is controlled by the ER-associated protein degradation (ERAD) pathway, which specifically targets misfolded proteins for degradation. Previously, we reported that HIV-1 replication is restricted in the human CD4(+) T cell line CEM.NKR (NKR). To understand this mechanism, we first analyzed cellular protein expression in NKR cells and discovered that levels of the mitochondrial translocator protein TSPO were upregulated by ∼64-fold. Notably, when NKR cells were treated with TSPO antagonist PK-11195, Ro5-4864, or diazepam, HIV restriction was completely disrupted, and TSPO knockdown by short hairpin RNAs (shRNAs) achieved a similar effect. We next analyzed viral protein expression, and, interestingly, we discovered that Env expression was specifically inhibited. Both TSPO knockdown and treatment with TSPO antagonist could restore Env expression in NKR cells. We further discovered that Env proteins were rapidly degraded and that kifunensine, an ERAD pathway inhibitor, could restore Env expression and viral replication, indicating that Env proteins were misfolded and degraded through the ERAD pathway in NKR cells. We also knocked out the TSPO gene in 293T cells using CRISPR/Cas9 (clustered, regularly interspaced, short palindromic repeat [CRISPR]/CRISPR-associated-9) technology and found that TSPO could similarly inhibit Env expression in these cells. Taken together, these results demonstrate that TSPO inhibits Env protein expression through the ERAD pathway and suggest that mitochondria play an important role in regulating the Env folding process. The HIV-1 Env glycoprotein is absolutely required for viral infection, and an understanding of its expression pathway in infected cells will identify new targets for antiretroviral therapies. Env proteins

  10. Proteomes and Ubiquitylomes Analysis Reveals the Involvement of Ubiquitination in Protein Degradation in Petunias1

    Science.gov (United States)

    Liu, Juanxu; Wei, Qian; Wang, Rongmin; Yang, Weiyuan; Ma, Yueyue; Chen, Guoju

    2017-01-01

    Petal senescence is a complex programmed process. It has been demonstrated previously that treatment with ethylene, a plant hormone involved in senescence, can extensively alter transcriptome and proteome profiles in plants. However, little is known regarding the impact of ethylene on posttranslational modification (PTM) or the association between PTM and the proteome. Protein degradation is one of the hallmarks of senescence, and ubiquitination, a major PTM in eukaryotes, plays important roles in protein degradation. In this study, we first obtained reference petunia (Petunia hybrida) transcriptome data via RNA sequencing. Next, we quantitatively investigated the petunia proteome and ubiquitylome and the association between them in petunia corollas following ethylene treatment. In total, 51,799 unigenes, 3,606 proteins, and 2,270 ubiquitination sites were quantified 16 h after ethylene treatment. Treatment with ethylene resulted in 14,448 down-regulated and 6,303 up-regulated unigenes (absolute log2 fold change > 1 and false discovery rate petunia. Several putative ubiquitin ligases were up-regulated at the protein and transcription levels. Our results showed that the global proteome and ubiquitylome were negatively correlated and that ubiquitination could be involved in the degradation of proteins during ethylene-mediated corolla senescence in petunia. Ethylene regulates hormone signaling transduction pathways at both the protein and ubiquitination levels in petunia corollas. In addition, our results revealed that ethylene increases the ubiquitination levels of proteins involved in endoplasmic reticulum-associated degradation. PMID:27810942

  11. Long-term aversive taste memory requires insular and amygdala protein degradation.

    Science.gov (United States)

    Rodriguez-Ortiz, Carlos J; Balderas, Israela; Saucedo-Alquicira, Fernando; Cruz-Castañeda, Paulina; Bermudez-Rattoni, Federico

    2011-03-01

    Some reports have shown that the ubiquitin-proteasome system (UPS) is necessary to degrade repressor factors to produce new proteins essential to memory consolidation. Furthermore, recent evidence suggests that memory updating also relies on protein degradation through the UPS. To evaluate whether degradation of proteins is part of the cellular events needed for long-term storage of taste aversion, we injected lactacystin--an UPS inhibitor--into the amygdala and/or insular cortex 30 min before the first or second training trials. The results revealed that degradation of proteins in either the amygdala or insular cortex suffices for long-term stabilization of first-time encounter taste aversion. On the other hand, lactacystin applied in the insula, but not in the amygdala, before the second training prevented long-term storage of updated information. Our results support that degradation of proteins by means of the UPS is required every time taste aversion is to be stored in long-term memory. Copyright © 2010 Elsevier Inc. All rights reserved.

  12. The effect of rumen ciliates on chitinolytic activity, chitin content and the number of fungal zoospores in the rumen fluid of sheep.

    Science.gov (United States)

    Miltko, Renata; Bełżecki, Grzegorz; Herman, Andrzej; Kowalik, Barbara; Skomiał, Jacek

    2016-12-01

    The objective of this study was to investigate the effect of selected protozoa on the degradation and concentration of chitin and the numbers of fungal zoospores in the rumen fluid of sheep. Three adult ewes were fed a hay-concentrate diet, defaunated, then monofaunated with Entodinium caudatum or Diploplastron affine alone and refaunated with natural rumen fauna. The average density of the protozoa population varied from 6.1 · 10(4) (D. affine) to 42.2 · 10(4) cells/ml rumen fluid (natural rumen fauna). The inoculation of protozoa in the rumen of defaunated sheep increased the total activity of chitinolytic enzymes from 2.9 to 3.6 μmol N-acetylglucosamine/g dry matter (DM) of rumen fluid per min, the chitin concentration from 6.3 to 7.2 mg/g DM of rumen fluid and the number of fungal zoospores from 8.1 to 10.9 · 10(5) cells/ml rumen fluid. All examined indices showed diurnal variations. Ciliate population density was highest immediately prior to feeding and lowest at 4 h thereafter. The opposite effects were observed for the numbers of fungal zoospores, the chitin concentration and chitinolytic activity. Furthermore, it was found that chitin from zoospores may account for up to 95% of total microbial chitin in the rumen fluid of sheep. In summary, the examined ciliate species showed the ability of chitin degradation as well as a positive influence on the development of the ruminal fungal population.

  13. Monoubiquitination of Tob/BTG family proteins competes with degradation-targeting polyubiquitination

    Energy Technology Data Exchange (ETDEWEB)

    Suzuki, Toru, E-mail: toru@ims.u-tokyo.ac.jp [Division of Oncology, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Kim, Minsoo [Division of Bacterial Infection, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Kozuka-Hata, Hiroko [Medical Proteomics Laboratory, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Watanabe, Masato [Department of Medical Genome Science, School of Frontier Sciences, University of Tokyo, 5-1-5 Kashiwanoha, Kashiwa 277-8562 (Japan); Oyama, Masaaki [Medical Proteomics Laboratory, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Tsumoto, Kouhei [Medical Proteomics Laboratory, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Department of Medical Genome Science, School of Frontier Sciences, University of Tokyo, 5-1-5 Kashiwanoha, Kashiwa 277-8562 (Japan); Yamamoto, Tadashi, E-mail: tyamamot@ims.u-tokyo.ac.jp [Division of Oncology, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Cell Signal Unit, Okinawa Institute of Science and Technology, 1919-1 Onna-son, Kunigami, Okinawa 904-0412 (Japan)

    2011-05-27

    Highlights: {yields} Tob/BTG family proteins are monoubiquitinated in the absence of E3s in vitro. {yields} Monoubiquitination sites of Tob are identified by mass spectrometry. {yields} The monoubiquitination event correlates with lower levels of polyubiquitination. -- Abstract: Tob belongs to the anti-proliferative Tob/BTG protein family. The expression level of Tob family proteins is strictly regulated both transcriptionally and through post-translational modification. Ubiquitin (Ub)/proteosome-dependent degradation of Tob family proteins is critical in controlling cell cycle progression and DNA damage responses. Various Ub ligases (E3s) are responsible for degradation of Tob protein. Here, we show that Tob family proteins undergo monoubiquitination even in the absence of E3s in vitro. Determination of the ubiquitination site(s) in Tob by mass spectrometric analysis revealed that two lysine residues (Lys48 and Lys63) located in Tob/BTG homology domain are ubiquitinated. A mutant Tob, in which both Lys48 and Lys63 are substituted with alanine, is more strongly polyubiquitinated than wild-type Tob in vivo. These data suggest that monoubiquitination of Tob family proteins confers resistance against polyubiquitination, which targets proteins for degradation. The strategy for regulating the stability of Tob family proteins suggests a novel role for monoubiquitination.

  14. Monoubiquitination of Tob/BTG family proteins competes with degradation-targeting polyubiquitination

    International Nuclear Information System (INIS)

    Suzuki, Toru; Kim, Minsoo; Kozuka-Hata, Hiroko; Watanabe, Masato; Oyama, Masaaki; Tsumoto, Kouhei; Yamamoto, Tadashi

    2011-01-01

    Highlights: → Tob/BTG family proteins are monoubiquitinated in the absence of E3s in vitro. → Monoubiquitination sites of Tob are identified by mass spectrometry. → The monoubiquitination event correlates with lower levels of polyubiquitination. -- Abstract: Tob belongs to the anti-proliferative Tob/BTG protein family. The expression level of Tob family proteins is strictly regulated both transcriptionally and through post-translational modification. Ubiquitin (Ub)/proteosome-dependent degradation of Tob family proteins is critical in controlling cell cycle progression and DNA damage responses. Various Ub ligases (E3s) are responsible for degradation of Tob protein. Here, we show that Tob family proteins undergo monoubiquitination even in the absence of E3s in vitro. Determination of the ubiquitination site(s) in Tob by mass spectrometric analysis revealed that two lysine residues (Lys48 and Lys63) located in Tob/BTG homology domain are ubiquitinated. A mutant Tob, in which both Lys48 and Lys63 are substituted with alanine, is more strongly polyubiquitinated than wild-type Tob in vivo. These data suggest that monoubiquitination of Tob family proteins confers resistance against polyubiquitination, which targets proteins for degradation. The strategy for regulating the stability of Tob family proteins suggests a novel role for monoubiquitination.

  15. The F-box protein FBXO44 mediates BRCA1 ubiquitination and degradation.

    Science.gov (United States)

    Lu, Yunzhe; Li, Jiezhi; Cheng, Dongmei; Parameswaran, Balaji; Zhang, Shaohua; Jiang, Zefei; Yew, P Renee; Peng, Junmin; Ye, Qinong; Hu, Yanfen

    2012-11-30

    BRCA1 mutations account for a significant proportion of familial breast and ovarian cancers. In addition, reduced BRCA1 protein is associated with sporadic cancer cases in these tissues. At the cellular level, BRCA1 plays a critical role in multiple cellular functions such as DNA repair and cell cycle checkpoint control. Its protein level is regulated in a cell cycle-dependent manner. However, regulation of BRCA1 protein stability is not fully understood. Our earlier study showed that the amino terminus of BRCA1 harbors a degron sequence that is sufficient and necessary for conferring BRCA1 degradation. In the current study, we used mass spectrometry to identify Skp1 that regulates BRCA1 protein stability. Small interfering RNA screening that targets all human F-box proteins uncovered FBXO44 as an important protein that influences BRCA1 protein level. The Skp1-Cul1-F-box-protein44 (SCF(FBXO44)) complex ubiquitinates full-length BRCA1 in vitro. Furthermore, the N terminus of BRCA1 mediates the interaction between BRCA1 and FBXO44. Overexpression of SCF(FBXO44) reduces BRCA1 protein level. Taken together, our work strongly suggests that SCF(FBXO44) is an E3 ubiquitin ligase responsible for BRCA1 degradation. In addition, FBXO44 expression pattern in breast carcinomas suggests that SCF(FBXO44)-mediated BRCA1 degradation might contribute to sporadic breast tumor development.

  16. The F-box Protein FBXO44 Mediates BRCA1 Ubiquitination and Degradation*

    Science.gov (United States)

    Lu, Yunzhe; Li, Jiezhi; Cheng, Dongmei; Parameswaran, Balaji; Zhang, Shaohua; Jiang, Zefei; Yew, P. Renee; Peng, Junmin; Ye, Qinong; Hu, Yanfen

    2012-01-01

    BRCA1 mutations account for a significant proportion of familial breast and ovarian cancers. In addition, reduced BRCA1 protein is associated with sporadic cancer cases in these tissues. At the cellular level, BRCA1 plays a critical role in multiple cellular functions such as DNA repair and cell cycle checkpoint control. Its protein level is regulated in a cell cycle-dependent manner. However, regulation of BRCA1 protein stability is not fully understood. Our earlier study showed that the amino terminus of BRCA1 harbors a degron sequence that is sufficient and necessary for conferring BRCA1 degradation. In the current study, we used mass spectrometry to identify Skp1 that regulates BRCA1 protein stability. Small interfering RNA screening that targets all human F-box proteins uncovered FBXO44 as an important protein that influences BRCA1 protein level. The Skp1-Cul1-F-box-protein44 (SCFFBXO44) complex ubiquitinates full-length BRCA1 in vitro. Furthermore, the N terminus of BRCA1 mediates the interaction between BRCA1 and FBXO44. Overexpression of SCFFBXO44 reduces BRCA1 protein level. Taken together, our work strongly suggests that SCFFBXO44 is an E3 ubiquitin ligase responsible for BRCA1 degradation. In addition, FBXO44 expression pattern in breast carcinomas suggests that SCFFBXO44-mediated BRCA1 degradation might contribute to sporadic breast tumor development. PMID:23086937

  17. Crude protein changes on grassland along a degradation gradient ...

    African Journals Online (AJOL)

    The aim with this investigation was to quantify the impact of different veld condition classes, viz. poor, moderate and good on soil-water utilization (SWU: crude protein produced per unit of evapotranspiration), during four growing seasons (2000/01 – 2003/2004). Evapotranspiration was determined by quantifying the ...

  18. Crude protein changes on grassland along a degradation gradient ...

    African Journals Online (AJOL)

    Evapotranspiration was determined by quantifying the soil-water balance equation with the aid of runoff plots and soil-water content measurements. Crude protein ... The study shows that it is important to keep grassland in optimal condition to utilize limited soil water for sustainable plant and therefore animal production.

  19. Short communication In situ ruminal degradability of protein feeds ...

    African Journals Online (AJOL)

    2016-12-28

    Dec 28, 2016 ... ISSN 0375-1589 (print), ISSN 2221-4062 (online) ... protein at a passage rate of 2% per hour was higher in meals than in grains. .... beef cattle reduced the consumption rate, but improved the feed conversion rate, increasing.

  20. Evaluation of feeds from tropical origin for in vitro methane production potential and rumen fermentation in vitro

    Directory of Open Access Journals (Sweden)

    Kaushik Pal

    2015-09-01

    Full Text Available Enteric methane arising due to fermentation of feeds in the rumen contributes substantially to the greenhouse gas emissions. Thus, like evaluation of chemical composition and nutritive values of feeds, methane production potential of each feed should be determined. This experiment was conducted to evaluate several feeds for methane production potential and rumen fermentation using in vitro gas production technique so that low methane producing feeds could be utilized to feed ruminants. Protein- and energy-rich concentrates (n=11, cereal and grass forages (n=11, and different straws and shrubs (n=12, which are commonly fed to ruminants in India, were collected from a number of locations. Gas production kinetics, methane production, degradability and rumen fermentation greatly varied (p<0.01 among feeds depending upon the chemical composition. Methane production (mL/g of degraded organic matter was lower (p<0.01 for concentrate than forages, and straws and shrubs. Among shrubs and straws, methane production was lower (p<0.01 for shrubs than straws. Methane production was correlated (p<0.05 with concentrations of crude protein (CP, ether extract and non-fibrous carbohydrate (NFC negatively, and with neutral detergent (NDF and acid detergent fiber (ADF positively. Potential gas production was negatively correlated (p=0.04 with ADF, but positively (p<0.01 with NFC content. Rate of gas production and ammonia concentration were influenced by CP content positively (p<0.05, but by NDF and ADF negatively (p<0.05. Total volatile fatty acid concentration and organic matter degradability were correlated (p<0.05 positively with CP and NFC content, but negatively with NDF and ADF content. The results suggest that incorporation of concentrates and shrubs replacing straws and forages in the diets of ruminants may decrease methane production.

  1. Evaluation of feeds from tropical origin for in vitro methane production potential and rumen fermentation in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Pal, K.; Patra, A. K.; Sahoo, K.

    2015-07-01

    Enteric methane arising due to fermentation of feeds in the rumen contributes substantially to the greenhouse gas emissions. Thus, like evaluation of chemical composition and nutritive values of feeds, methane production potential of each feed should be determined. This experiment was conducted to evaluate several feeds for methane production potential and rumen fermentation using in vitro gas production technique so that low methane producing feeds could be utilized to feed ruminants. Protein- and energy-rich concentrates (n=11), cereal and grass forages (n=11), and different straws and shrubs (n=12), which are commonly fed to ruminants in India, were collected from a number of locations. Gas production kinetics, methane production, degradability and rumen fermentation greatly varied (p<0.01) among feeds depending upon the chemical composition. Methane production (mL/g of degraded organic matter) was lower (p<0.01) for concentrate than forages, and straws and shrubs. Among shrubs and straws, methane production was lower (p<0.01) for shrubs than straws. Methane production was correlated (p<0.05) with concentrations of crude protein (CP), ether extract and non-fibrous carbohydrate (NFC) negatively, and with neutral detergent (NDF) and acid detergent fiber (ADF) positively. Potential gas production was negatively correlated (p=0.04) with ADF, but positively (p<0.01) with NFC content. Rate of gas production and ammonia concentration were influenced by CP content positively (p<0.05), but by NDF and ADF negatively (p<0.05). Total volatile fatty acid concentration and organic matter degradability were correlated (p<0.05) positively with CP and NFC content, but negatively with NDF and ADF content. The results suggest that incorporation of concentrates and shrubs replacing straws and forages in the diets of ruminants may decrease. (Author)

  2. Variation in ruminal in situ degradation of crude protein and starch from maize grains compared to in vitro gas production kinetics and physical and chemical characteristics.

    Science.gov (United States)

    Seifried, Natascha; Steingaß, Herbert; Schipprack, Wolfgang; Rodehutscord, Markus

    2016-10-01

    The objectives of this study were (1) to evaluate in situ ruminal dry matter (DM), crude protein (CP) and starch degradation characteristics and in vitro gas production (GP) kinetics using a set of 20 different maize grain genotypes and (2) to predict the effective degradation (ED) of CP and starch from chemical and physical characteristics alone or in combination with in vitro GP measurements. Maize grains were characterised by different chemical and physical characteristics. Ruminal in situ degradation was measured in three lactating Jersey cows. Ground grains (sieve size: 2 mm) were incubated in bags for 1, 2, 4, 8, 16, 24, 48 and 72 h. Bag residues were analysed for CP and starch content. Degradation kinetics was determined and the ED of DM, CP and starch calculated using a ruminal passage rate of 5%/h and 8%/h. The GP of the grains (sieve size: 1 mm) was recorded after 2, 4, 6, 8, 12, 24, 48 and 72 h incubation in buffered rumen fluid and fitted to an exponential equation to determine GP kinetics. Correlations and stepwise multiple linear regressions were evaluated for the prediction of ED calculated for a passage rate of 5%/h (ED5) for CP (EDCP5) and starch (EDST5). The in situ parameters and ED5 varied widely between genotypes with average values (±SD) of 64% ± 4.2, 62% ± 4.1 and 65% ± 5.2 for ED5 of DM, EDCP5 and EDST5 and were on average 10 percentage points lower for a passage rate of 8%/h. Degradation rates varied between 4.8%/h and 7.4%/h, 4.1%/h and 6.5%/h and 5.3%/h and 8.9%/h for DM, CP and starch, respectively. These rates were in the same range as GP rates (6.0-8.3%/h). The EDCP5 and EDST5 were related to CP concentration and could be evaluated in detail using CP fractions and specific amino acids. In vitro GP measurements and GP rates correlated well with EDCP5 and EDST5 and predicted EDCP5 and EDST5 in combination with the chemical characteristics of the samples. Equations can be used to obtain quick and cost effective information

  3. Consumo e dinâmica ruminal da fibra em detergente neutro em bovinos em pastejo no período das águas recebendo suplementação com nitrogênio não-proteico e/ou proteína verdadeira Intake and rumen dynamics of neutral detergent fiber in grazing cattle supplemented with non-protein nitrogen and, or true protein during the rainy season

    Directory of Open Access Journals (Sweden)

    Viviane Aparecida Carli Costa

    2011-12-01

    during rainy season. Five crossbred Holstein × Zebu steers, averaging 335±35 kg of body weight and fitted with rumen and abomasum canullaes were used. The treatments were: control (only pasture, and supplements based on urea, 2/3 of nitrogenous compounds from urea and 1/3 of nitrogenous compounds from albumin, 1/3 of nitrogenous compounds from urea and 2/3 of nitrogenous compounds from albumin, and albumin. Two hundred grams/d of crude protein (CP were supplied from supplements. The experiment was carried out according to a 5 × 5 Latin square design, with five 15-day experimental periods. There were no effects of supplementation on voluntary intake, except for CP intake, which was increased by supplementation. The replacement of urea by albumin in the supplements caused linear effect on the CP intake. The intakes of the other diet components were not affected by the supplement composition. There was no effect on ruminal rate of passage of fibrous compounds. Supplementation increased the estimates of common rate of lag and degradation of NDF. However, no effect of supplement composition alteration was observed on this parameter. Supplementation of cattle with rumen degradable (protein or non-protein nitrogenous compounds for grazing cattle during rainy season does not affect voluntary intake of pasture.

  4. Development of Simple and Precise Method of Arginine Determination in Rumen Fluid by Spectrophotometer

    International Nuclear Information System (INIS)

    Chacher, B.; Marghazani, I. B.; Liu, J. X.; Liu, H. Y.

    2015-01-01

    The objective of current study was to build up a convenient, economic and accurate procedure to determine arginine (ARG) concentration in rumen fluid. Rumen fluid was collected from 3 rumen fistulated Chinese Holstein dairy cows and added with or without (control) 1mmol/l unprotected ARG and blank (with only medium) in to syringe system in triplicate as a replicate. All syringes were incubated in water bath at 39 Degree C for 0, 2, 4, 6, 12 and 24 h and were terminated to measure the ARG concentration. Sakaguchi reaction method was used to analyze the ARG concentration in rumen fluid by determining the rumen degradation rate of protected and unprotected ARG. Temperature, time and absorbance were optimized in the procedure based on Sakaguchi reaction. Color consistency remained 4-6 min. The optimum temperature (0-5) Degree C was observed for maximum optical density 0.663 at wave length 500 nm. Minimum ARG that could be determined in rumen fluid by spectrophotometer was 4-5 μ g/ml. No significance (P>0.05) difference were observed between two results derived from spectrophotometer and amino acid analyzer methods. In conclusion, the spectrophotometer method of ARG determination in rumen fluid based on Sakaguchi reaction is easy, accurate, and economical and could be useful in learning ARG metabolism in the rumen. (author)

  5. In vitro fermentation characteristics of diets with different forage/concentrate ratios: comparison of rumen and faecal inocula.

    Science.gov (United States)

    Zicarelli, Fabio; Calabrò, Serena; Cutrignelli, Monica I; Infascelli, Federico; Tudisco, Raffaella; Bovera, Fulvia; Piccolo, Vincenzo

    2011-05-01

    The aim of this trial was to evaluate the replacement of rumen fluid with faeces as inoculum in studying the in vitro fermentation characteristics of diets for ruminants using the in vitro gas production technique. Six iso-protein diets with different forage/concentrate ratios were incubated with rumen fluid (RI) or faeces (FI) collected from sheep. Most of the fermentation parameters were influenced by diet and inoculum (P < 0.01). With both inocula, organic matter degradability (dOM), cumulative gas production (OMCV) and maximum fermentation rate (R(max) ) increased as the amount of concentrate in the diet increased. R(max) was lower with FI vs RI (P < 0.01); dOM was higher with FI vs RI and the diet × inoculum interaction was significant. As expected, with both inocula, R(max) increased as the neutral detergent fibre content of the diet decreased. Significant correlations were obtained using both inocula between OMCV/dOM and gas/volatile fatty acid (VFA), while the correlation VFA/dOM was significant only with FI. The microbial biomass yield calculated by stoichiometric analysis for all diets was higher with FI vs RI. With FI the organic matter used for microbial growth showed an overall decreasing trend as the amount of concentrate in the diet increased. The results indicate that both faeces and rumen fluid from sheep have the potential to be used as inoculum for the in vitro gas production technique. Copyright © 2011 Society of Chemical Industry.

  6. Identifiability study of the proteins degradation model, based on ADM1, using simultaneous batch experiments

    DEFF Research Database (Denmark)

    Flotats, X.; Palatsi, J.; Ahring, Birgitte Kiær

    2006-01-01

    are not inhibiting the hydrolysis process. The ADM1 model adequately expressed the consecutive steps of hydrolysis and acidogenesis, with estimated kinetic values corresponding to a fast acidogenesis and slower hydrolysis. The hydrolysis was found to be the rate limiting step of anaerobic degradation. Estimation...... of yield coefficients based on the relative initial slopes of VFA profiles obtained in a simple batch experiment produced satisfactory results. From the identification study, it was concluded that it is possible to determine univocally the related kinetic parameter values for protein degradation...... if the evolution of amino acids is measured in simultaneous batch experiments, with different initial protein and amino acids concentrations....

  7. Protein nutrition of growing cattle

    International Nuclear Information System (INIS)

    Chalupa, W.; Scott, G.C.

    1976-01-01

    In vitro studies on apparent degradation of amino acids by mixed and pure cultures of rumen bacteria demonstrated that (a) amino acids are degraded at differing rates (Arg, Thr>Lys, Phe, Leu, Ile>Val, Met); (b) certain amino acids (Met, Val, Try, Orn) are degraded to greater extents when fermented alone than in conjunction with other amino acids; (c) individual strains of rumen bacteria do not utilize all amino acids; and (d) total ruminal degradation of amino acids is the result of extensive bacterial interaction, and may vary greatly depending on the predominant types of micro-organisms present. Abomasal infusion of a mixture of 10 essential amino acids consistently increased nitrogen retention, but attempts to elucidate primary limiting amino acids were not conclusive. Our data suggested that supplementary methionine alone may not significantly increase nitrogen retention, but methionine must be present in order to obtain responses from other amino acids. Methionine plus lysine plus threonine usually increased nitrogen retention, but the magnitude of responses varied. The classical nitrogen balance technique may lack the sensitivity needed to detect small responses resulting from supplements of single amino acids, or growing cattle, unlike sheep used for wool growth, may not be suffering from specific amino acid deficiencies. Chemical suppression of ruminal degradation of amino acids produced significant increases in nitrogen retention and growth, and improved feed efficiencies. Productivity responses to rumen bypass techniques would seem to depend primarily upon (a) the degree to which dietary protein is degraded in the rumen, and (b) the quantity of absorbable amino acids supplied by the diet in relation to quantities required by the animal. (author)

  8. Rumen-protected choline: A significance effect on dairy cattle nutrition.

    Science.gov (United States)

    Jayaprakash, G; Sathiyabarathi, M; Robert, M Arokia; Tamilmani, T

    2016-08-01

    Choline is a vitamin-like substance it has multi-function in animal production, reproduction, and health. The transition period is most crucial stage in lactation cycle of dairy cows due to its association with negative hormonal and energy balances. Unfortunately, unprotected choline easily degrades in the rumen; therefore, choline added to the diet in a rumen-protected form. The use of rumen-protected choline (RPC) is a preventive measurement for the fatty liver syndrome and ketosis; may improve milk production as well as milk composition and reproduction parameters. This review summarizes the effectiveness of RPC on animal production, health, and reproduction.

  9. Study of Local Herb Potency as Rumen Modifier: The Effect of Red Ginger (Zingiber officinale Var.Rubrum) on Parameters of Ruminal Fermentation In Vitro

    Science.gov (United States)

    Kurniawati, A.; Widodo; Artama, W. T.; Yusiati, L. M.

    2018-02-01

    Essential oil is one of rumen modifier alternatives due to its antimicrobial property. Red ginger is one of local herbs with high essential oil content. The effect of red ginger on rumen fermentation parameters was studied in this research using in vitro gas production method. Five level of red ginger meal was added to the diet to meet final essential oil concentration in fermentation medium of 0, 25, 50, 75 and 100 mg/L. Substrate of fermentation as microbial feed composed of Penisetum hybride, rice bran, and wheat pollard in ratio 60:20:20 DM basis. Fermentation was carried out for 24 h at 39°C. Total gas production was measured at the end of incubation and sample for methane analysis was taken. Medium sample was taken for analysis of pH, ammonium and VFA concentration, microbial protein and protozoa number. Data showed that addition of red ginger in the diet did not affect the pH, ammonia and VFA concentration, microbial protein and also protozoa number. However, red ginger addition significantly decrease ammonia concentration in all treatment. It could be concluded that addition of red ginger in the diet reduced degradation protein in the rumen as illustrated in lower ammonia concentration.

  10. ESCRT-dependent degradation of ubiquitylated plasma membrane proteins in plants.

    Science.gov (United States)

    Isono, Erika; Kalinowska, Kamila

    2017-12-01

    To control the abundance of plasma membrane receptors and transporters is crucial for proper perception and response to extracellular signals from surrounding cells and the environment. Posttranslational modification of plasma membrane proteins, especially ubiquitin conjugation or ubiquitylation, is key for the determination of stability for many transmembrane proteins localized on the cell surface. The targeted degradation is ensured by a complex network of proteins among which the endosomal sorting complex required for transport (ESCRT) plays a central role. This review focuses on progresses made in recent years on the understanding of the function of the ESCRT machinery in the degradation of ubiquitylated plasma membrane proteins in plants. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Iron-Binding Protein Degradation by Cysteine Proteases of Naegleria fowleri

    Directory of Open Access Journals (Sweden)

    Moisés Martínez-Castillo

    2015-01-01

    Full Text Available Naegleria fowleri causes acute and fulminant primary amoebic meningoencephalitis. This microorganism invades its host by penetrating the olfactory mucosa and then traveling up the mesaxonal spaces and crossing the cribriform plate; finally, the trophozoites invade the olfactory bulbs. During its invasion, the protozoan obtains nutrients such as proteins, lipids, carbohydrates, and cationic ions (e.g., iron, calcium, and sodium from the host. However, the mechanism by which these ions are obtained, particularly iron, is poorly understood. In the present study, we evaluated the ability of N. fowleri to degrade iron-binding proteins, including hololactoferrin, transferrin, ferritin, and hemoglobin. Zymography assays were performed for each substrate under physiological conditions (pH 7 at 37°C employing conditioned medium (CM and total crude extracts (TCEs of N. fowleri. Different degradation patterns with CM were observed for hololactoferrin, transferrin, and hemoglobin; however, CM did not cause ferritin degradation. In contrast, the TCEs degraded only hololactoferrin and transferrin. Inhibition assays revealed that cysteine proteases were involved in this process. Based on these results, we suggest that CM and TCEs of N. fowleri degrade iron-binding proteins by employing cysteine proteases, which enables the parasite to obtain iron to survive while invading the central nervous system.

  12. Molecular chaperones in targeting misfolded proteins for ubiquitin-dependent degradation

    DEFF Research Database (Denmark)

    Kriegenburg, Franziska; Ellgaard, Lars; Hartmann-Petersen, Rasmus

    2012-01-01

    The accumulation of misfolded proteins presents a considerable threat to the health of individual cells and has been linked to severe diseases, including neurodegenerative disorders. Considering that, in nature, cells often are exposed to stress conditions that may lead to aberrant protein...... conformational changes, it becomes clear that they must have an efficient quality control apparatus to refold or destroy misfolded proteins. In general, cells rely on molecular chaperones to seize and refold misfolded proteins. If the native state is unattainable, misfolded proteins are targeted for degradation...... via the ubiquitin-proteasome system. The specificity of this proteolysis is generally provided by E3 ubiquitin-protein ligases, hundreds of which are encoded in the human genome. However, rather than binding the misfolded proteins directly, most E3s depend on molecular chaperones to recognize...

  13. Comparison of Rumen and Manure Microbiomes and Implications for the Inoculation of Anaerobic Digesters

    OpenAIRE

    Emine Gozde Ozbayram; Orhan Ince; Bahar Ince; Hauke Harms; Sabine Kleinsteuber

    2018-01-01

    Cattle manure is frequently used as an inoculum for the start-up of agricultural biogas plants or as a co-substrate in the anaerobic digestion of lignocellulosic feedstock. Ruminal microbiota are considered to be effective plant fiber degraders, but the microbes contained in manure do not necessarily reflect the rumen microbiome. The aim of this study was to compare the microbial community composition of cow rumen and manure with respect to plant fiber-digesting microbes. Bacterial and methan...

  14. Reduced synaptic vesicle protein degradation at lysosomes curbs TBC1D24/sky-induced neurodegeneration.

    Science.gov (United States)

    Fernandes, Ana Clara; Uytterhoeven, Valerie; Kuenen, Sabine; Wang, Yu-Chun; Slabbaert, Jan R; Swerts, Jef; Kasprowicz, Jaroslaw; Aerts, Stein; Verstreken, Patrik

    2014-11-24

    Synaptic demise and accumulation of dysfunctional proteins are thought of as common features in neurodegeneration. However, the mechanisms by which synaptic proteins turn over remain elusive. In this paper, we study Drosophila melanogaster lacking active TBC1D24/Skywalker (Sky), a protein that in humans causes severe neurodegeneration, epilepsy, and DOOR (deafness, onychdystrophy, osteodystrophy, and mental retardation) syndrome, and identify endosome-to-lysosome trafficking as a mechanism for degradation of synaptic vesicle-associated proteins. In fly sky mutants, synaptic vesicles traveled excessively to endosomes. Using chimeric fluorescent timers, we show that synaptic vesicle-associated proteins were younger on average, suggesting that older proteins are more efficiently degraded. Using a genetic screen, we find that reducing endosomal-to-lysosomal trafficking, controlled by the homotypic fusion and vacuole protein sorting (HOPS) complex, rescued the neurotransmission and neurodegeneration defects in sky mutants. Consistently, synaptic vesicle proteins were older in HOPS complex mutants, and these mutants also showed reduced neurotransmission. Our findings define a mechanism in which synaptic transmission is facilitated by efficient protein turnover at lysosomes and identify a potential strategy to suppress defects arising from TBC1D24 mutations in humans. © 2014 Fernandes et al.

  15. Protein degradation in preimplantation mouse embryos and the lethality of tritiated amino acids

    International Nuclear Information System (INIS)

    Wielbold, J.L.

    1982-01-01

    The role of protein degradation in preimplantation development in the mouse was studied. Proteins of morulae and blastocysts (M and B) cultured in vitro after labeling for 1 hour (h) in 3 H-leucine exhibit a mean half-life (t 1 / 2 ) of 8.1 h. The t 1 / 2 tends to increase (9.5 h) when 10% fetal calf serum is added to the chase medium. This decrease in protein degradation in the presence of serum is associated with an increase in the percentage of B that are hatching (P 3 H-leucine in their proteins than did Day 4 embryos remaining in culture (P<0.02), while Day 4 embryos in a Day 3 uterus retained the same amount of radioactivity as did Day 4 embryos in culture. This differential effect of uterine environment was also seen when Day 4 embryos were transferred to recipients. More fetuses developed to term when the recipient was in Day 3 of PSP (50.8%) than when the recipient was in Day 4 PSP (25.9%, P<0.001), regardless of the age of the recipient. Age of the recipient does affect the percentage of transferred embryos developing to term. Thus, protein degradation may vary with the stage of embryo development and the conditions to which the embryos are exposed. However, even low levels of incorporated tritiated leucine can have lethal effects on the embryos and compromise the validity of the protein half-lives determined

  16. Cellular Cholesterol Regulates Ubiquitination and Degradation of the Cholesterol Export Proteins ABCA1 and ABCG1*

    Science.gov (United States)

    Hsieh, Victar; Kim, Mi-Jurng; Gelissen, Ingrid C.; Brown, Andrew J.; Sandoval, Cecilia; Hallab, Jeannette C.; Kockx, Maaike; Traini, Mathew; Jessup, Wendy; Kritharides, Leonard

    2014-01-01

    The objective of this study was to examine the influence of cholesterol in post-translational control of ABCA1 and ABCG1 protein expression. Using CHO cell lines stably expressing human ABCA1 or ABCG1, we observed that the abundance of these proteins is increased by cell cholesterol loading. The response to increased cholesterol is rapid, is independent of transcription, and appears to be specific for these membrane proteins. The effect is mediated through cholesterol-dependent inhibition of transporter protein degradation. Cell cholesterol loading similarly regulates degradation of endogenously expressed ABCA1 and ABCG1 in human THP-1 macrophages. Turnover of ABCA1 and ABCG1 is strongly inhibited by proteasomal inhibitors and is unresponsive to inhibitors of lysosomal proteolysis. Furthermore, cell cholesterol loading inhibits ubiquitination of ABCA1 and ABCG1. Our findings provide evidence for a rapid, cholesterol-dependent, post-translational control of ABCA1 and ABCG1 protein levels, mediated through a specific and sterol-sensitive mechanism for suppression of transporter protein ubiquitination, which in turn decreases proteasomal degradation. This provides a mechanism for acute fine-tuning of cholesterol transporter activity in response to fluctuations in cell cholesterol levels, in addition to the longer term cholesterol-dependent transcriptional regulation of these genes. PMID:24500716

  17. The Polerovirus silencing suppressor P0 targets ARGONAUTE proteins for degradation.

    Science.gov (United States)

    Baumberger, Nicolas; Tsai, Ching-Hsui; Lie, Miranda; Havecker, Ericka; Baulcombe, David C

    2007-09-18

    Plant and animal viruses encode suppressor proteins of an adaptive immunity mechanism in which viral double-stranded RNA is processed into 21-25 nt short interfering (si)RNAs. The siRNAs guide ARGONAUTE (AGO) proteins so that they target viral RNA. Most viral suppressors bind long dsRNA or siRNAs and thereby prevent production of siRNA or binding of siRNA to AGO. The one exception is the 2b suppressor of Cucumoviruses that binds to and inhibits AGO1. Here we describe a novel suppressor mechanism in which a Polerovirus-encoded F box protein (P0) targets the PAZ motif and its adjacent upstream sequence in AGO1 and mediates its degradation. F box proteins are components of E3 ubiquitin ligase complexes that add polyubiquitin tracts on selected lysine residues and thereby mark a protein for proteasome-mediated degradation. With P0, however, the targeted degradation of AGO is insensitive to inhibition of the proteasome, indicating that the proteasome is not involved. We also show that P0 does not block a mobile signal of silencing, indicating that the signal molecule does not have AGO protein components. The ability of P0 to block silencing without affecting signal movement may contribute to the phloem restriction of viruses in the Polerovirus group.

  18. Fabrication of Flexible, Fully Organic, Degradable Energy Storage Devices Using Silk Proteins.

    Science.gov (United States)

    Pal, Ramendra K; Kundu, Subhas C; Yadavalli, Vamsi K

    2018-03-21

    Flexible and thin-film devices are of great interest in epidermal and implantable bioelectronics. The integration of energy storage and delivery devices such as supercapacitors (SCs) with properties such as flexibility, miniaturization, biocompatibility, and degradability are sought for such systems. Reducing e-waste and using sustainable materials and processes are additional desirable qualities. Herein, a silk protein-based biocompatible and degradable thin-film microSC (μSC) is reported. A protein carrier with the conducting polymer poly(3,4-ethylenedioxythiophene) polystyrene sulfonate and reduced graphene oxide dopant is used as a photopatternable biocomposite ink. Active electrodes are fabricated using photolithography under benign conditions, using only water as the solvent. These electrodes are printed on flexible protein sheets to form degradable, organic devices with a benign agarose-NaCl gel electrolyte. High capacitance, power density, cycling stability over 500 cycles, and the ability to power a light-emitting diode are shown. The device is flexible, can sustain cyclic mechanical stresses over 450 cycles, and retain capacitive properties over several days in liquid. Significantly, the μSCs are cytocompatible and completely degraded over the period of ∼1 month. By precise control of the device configuration, these silk protein-based, all-polymer organic devices can be designed to be tunably transient and provide viable alternatives for powering flexible and implantable bioelectronics.

  19. In situ ruminal crude protein degradability of by-products from cereals, oilseeds and animal origin

    NARCIS (Netherlands)

    Habib, G.; Khan, N.A.; Ali, M.; Bezabih, M.

    2013-01-01

    The aim of this study was to establish a database on in situ ruminal crude protein (CP) degradability characteristics of by-products from cereal grains, oilseeds and animal origin commonly fed to ruminants in Pakistan and South Asian Countries. The oilseed by-products were soybean meal, sunflower

  20. Delaying aging and the aging-associated decline in protein homeostasis by inhibition of tryptophan degradation

    NARCIS (Netherlands)

    van der Goot, Annemieke T.; Zhu, Wentao; Vazquez-Manrique, Rafael P.; Seinstra, Renee I.; Dettmer, Katja; Michels, Helen; Farina, Francesca; Krijnen, Jasper; Melki, Ronald; Buijsman, Rogier C.; Silva, Mariana Ruiz; Thijssen, Karen L.; Kema, Ido P.; Neri, Christian; Oefner, Peter J.; Nollen, Ellen A. A.

    2012-01-01

    Toxicity of aggregation-prone proteins is thought to play an important role in aging and age-related neurological diseases like Parkinson and Alzheimer's diseases. Here, we identify tryptophan 2,3-dioxygenase (tdo-2), the first enzyme in the kynurenine pathway of tryptophan degradation, as a

  1. Degradation and de novo synthesis of D1 protein and psbA ...

    Indian Academy of Sciences (India)

    This shows that synthesis of D1 protein is not the only component involved in the recovery process. Our events, which ... transcript levels in the green alga Chlamydomonas reinhardtii in ..... and Gaba V 1996 Accelerated degradation of the D2 ...

  2. Differential expression in Phanerochaete chrysosporium of membrane- associated proteins relevant to lignin degradation

    Science.gov (United States)

    Semarjit Shary; Alexander N. Kapich; Ellen A. Panisko; Jon K. Magnuson; Daniel Cullen; Kenneth E. Hammel

    2008-01-01

    Fungal lignin-degrading systems likely include membrane-associated proteins that participate in diverse processes such as uptake and oxidation of lignin fragments, production of ligninolytic secondary metabolites, and defense of the mycelium against ligninolytic oxidants. Little is known about the nature or regulation of these membrane-associated components. We grew...

  3. Some aspects of nitrogen metabolism in the bovine rumen

    International Nuclear Information System (INIS)

    Nikolic, J.A.

    1976-01-01

    Studies on the use of urea as a source of nitrogen for microbial protein synthesis showed that utilization was more efficient when the protein-N content of the diet was low. However, when the total nitrogen of the dietary dry matter was reduced below 1.9%, there was a drop in the protein-N content of rumen dry matter and a reduction in the daily flow of protein through the duodenum. The mean post-prandial rumen ammonia concentrations were below 5mg NH 3 -N/100ml. In vitro work with 15 N-labelled urea and ammonia salts showed that the overall utilization rate of ammonia was not significantly affected by mean concentrations between 1.6 and 16.7mg/ml, but that net utilization was lower below 5.8mg/100ml. It is suggested that increased lysis of susceptible micro-organisms or increased proteolysis of feed proteins may account for these findings. The concentrations of ammonia-N, protein-N and volatile fatty acids in the rumen were not affected by increases in the amount of calcium, phosphorus, potassium or sulphur in the diet, although the concentrations of these elements were significantly increased in the rumen. A wide dietary Ca/P ratio (3.26) tended to reduce the stability of rumen contents leading to a low pH and foaming. Increases in mean sulphide concentration from 3.6 to 8.0mg/l had no effect on overall protein synthesis rate as indicated by the 35 S incorporation rate. (author)

  4. Omics insights into rumen ureolytic bacterial community and urea metabolism in dairy cows

    OpenAIRE

    Jin, Di

    2017-01-01

    Urea has been used in the diets of ruminants as a non-protein nitrogen source. Ureolytic bacteria are key organisms in the rumen producing urease enzymes to catalyze the breakdown of urea to ammonia (NH3), and the NH3 is used as nitrogen for microbial protein synthesis. In the rumen, hydrolysis of urea to NH3 occurs at a greater rate than NH3 can be utilized by rumen bacteria, and excess ammonia absorbed into blood may be harmful to the animals. Nowadays, little is known about the information...

  5. ERManI (Endoplasmic Reticulum Class I α-Mannosidase) Is Required for HIV-1 Envelope Glycoprotein Degradation via Endoplasmic Reticulum-associated Protein Degradation Pathway.

    Science.gov (United States)

    Zhou, Tao; Frabutt, Dylan A; Moremen, Kelley W; Zheng, Yong-Hui

    2015-09-04

    Previously, we reported that the mitochondrial translocator protein (TSPO) induces HIV-1 envelope (Env) degradation via the endoplasmic reticulum (ER)-associated protein degradation (ERAD) pathway, but the mechanism was not clear. Here we investigated how the four ER-associated glycoside hydrolase family 47 (GH47) α-mannosidases, ERManI, and ER-degradation enhancing α-mannosidase-like (EDEM) proteins 1, 2, and 3, are involved in the Env degradation process. Ectopic expression of these four α-mannosidases uncovers that only ERManI inhibits HIV-1 Env expression in a dose-dependent manner. In addition, genetic knock-out of the ERManI gene MAN1B1 using CRISPR/Cas9 technology disrupts the TSPO-mediated Env degradation. Biochemical studies show that HIV-1 Env interacts with ERManI, and between the ERManI cytoplasmic, transmembrane, lumenal stem, and lumenal catalytic domains, the catalytic domain plays a critical role in the Env-ERManI interaction. In addition, functional studies show that inactivation of the catalytic sites by site-directed mutagenesis disrupts the ERManI activity. These studies identify ERManI as a critical GH47 α-mannosidase in the ER-associated protein degradation pathway that initiates the Env degradation and suggests that its catalytic domain and enzymatic activity play an important role in this process. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  6. Determining the amount of rumen-protected methionine supplement that corresponds to the optimal levels of methionine in metabolizable protein for maximizing milk protein production and profit on dairy farms.

    Science.gov (United States)

    Cho, J; Overton, T R; Schwab, C G; Tauer, L W

    2007-10-01

    The profitability of feeding rumen-protected Met (RPMet) sources to produce milk protein was estimated using a 2-step procedure: First, the effect of Met in metabolizable protein (MP) on milk protein production was estimated by using a quadratic Box-Cox functional form. Then, using these estimation results, the amounts of RPMet supplement that corresponded to the optimal levels of Met in MP for maximizing milk protein production and profit on dairy farms were determined. The data used in this study were modified from data used to determine the optimal level of Met in MP for lactating cows in the Nutrient Requirements of Dairy Cattle (NRC, 2001). The data used in this study differ from that in the NRC (2001) data in 2 ways. First, because dairy feed generally contains 1.80 to 1.90% Met in MP, this study adjusts the reference production value (RPV) from 2.06 to 1.80 or 1.90%. Consequently, the milk protein production response is also modified to an RPV of 1.80 or 1.90% Met in MP. Second, because this study is especially interested in how much additional Met, beyond the 1.80 or 1.90% already contained in the basal diet, is required to maximize farm profits, the data used are limited to concentrations of Met in MP above 1.80 or 1.90%. This allowed us to calculate any additional cost to farmers based solely on the price of an RPMet supplement and eliminated the need to estimate the dollar value of each gram of Met already contained in the basal diet. Results indicated that the optimal level of Met in MP for maximizing milk protein production was 2.40 and 2.42%, where the RPV was 1.80 and 1.90%, respectively. These optimal levels were almost identical to the recommended level of Met in MP of 2.40% in the NRC (2001). The amounts of RPMet required to increase the percentage of Met in MP from each RPV to 2.40 and 2.42% were 21.6 and 18.5 g/d, respectively. On the other hand, the optimal levels of Met in MP for maximizing profit were 2.32 and 2.34%, respectively. The amounts

  7. Effect of starch fermentation in the rumen on voluntary intake of ...

    African Journals Online (AJOL)

    such as minerals, protein, or rumen ammonia were maintained at adequate ... The relationship between voluntary feed intake and starch infusion was tested for .... artificial saliva (Tilley & Terry, 1%3) before filtering according to the same in ...

  8. The central domain of yeast transcription factor Rpn4 facilitates degradation of reporter protein in human cells.

    Science.gov (United States)

    Morozov, A V; Spasskaya, D S; Karpov, D S; Karpov, V L

    2014-10-16

    Despite high interest in the cellular degradation machinery and protein degradation signals (degrons), few degrons with universal activity along species have been identified. It has been shown that fusion of a target protein with a degradation signal from mammalian ornithine decarboxylase (ODC) induces fast proteasomal degradation of the chimera in both mammalian and yeast cells. However, no degrons from yeast-encoded proteins capable to function in mammalian cells were identified so far. Here, we demonstrate that the yeast transcription factor Rpn4 undergoes fast proteasomal degradation and its central domain can destabilize green fluorescent protein and Alpha-fetoprotein in human HEK 293T cells. Furthermore, we confirm the activity of this degron in yeast. Thus, the Rpn4 central domain is an effective interspecies degradation signal. Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  9. Mangosteen peel can reduce methane production and rumen ...

    African Journals Online (AJOL)

    TROPIKA 18

    2016-11-24

    Nov 24, 2016 ... especially total protozoa and total methanogens. ..... (10−1/slope − 1) × 100 ..... http://www.fao.org/docrep/012/k7930e/k7930e00.pdf. .... on rumen ecology, microbial protein synthesis, digestibility and voluntary feed intake in ...

  10. Pathogenic prion protein is degraded by a manganese oxide mineral found in soils

    Science.gov (United States)

    Russo, F.; Johnson, C.J.; McKenzie, D.; Aiken, Judd M.; Pedersen, J.A.

    2009-01-01

    Prions, the aetiological agents of transmissible spongiform encephalopathies, exhibit extreme resistance to degradation. Soil can retain prion infectivity in the environment for years. Reactive soil components may, however, contribute to the inactivation of prions in soil. Members of the birnessite family of manganese oxides (MnO2) rank among the strongest natural oxidants in soils. Here, we report the abiotic degradation of pathogenic prion protein (PrPTSE) by a synthetic analogue of naturally occurring birnessite minerals. Aqueous MnO2 suspensions degraded the PrPTSE as evidenced by decreased immunoreactivity and diminished ability to seed protein misfolding cyclic amplification reactions. Birnessite-mediated PrPTSE degradation increased as a solution's pH decreased, consistent with the pH-dependence of the redox potential of MnO2. Exposure to 5.6 mg MnO2 ml-1 (PrPTSE:MnO2=1 : 110) decreased PrPTSE levels by ???4 orders of magnitude. Manganese oxides may contribute to prion degradation in soil environments rich in these minerals. ?? 2009 SGM.

  11. HIV-1 accessory proteins VPR and Vif modulate antiviral response by targeting IRF-3 for degradation

    International Nuclear Information System (INIS)

    Okumura, Atsushi; Alce, Tim; Lubyova, Barbora; Ezelle, Heather; Strebel, Klaus; Pitha, Paula M.

    2008-01-01

    The activation of IRF-3 during the early stages of viral infection is critical for the initiation of the antiviral response; however the activation of IRF-3 in HIV-1 infected cells has not yet been characterized. We demonstrate that the early steps of HIV-1 infection do not lead to the activation and nuclear translocation of IRF-3; instead, the relative levels of IRF-3 protein are decreased due to the ubiquitin-associated proteosome degradation. Addressing the molecular mechanism of this effect we show that the degradation is independent of HIV-1 replication and that virion-associated accessory proteins Vif and Vpr can independently degrade IRF-3. The null mutation of these two genes reduced the capacity of the HIV-1 virus to down modulate IRF-3 levels. The degradation was associated with Vif- and Vpr-mediated ubiquitination of IRF-3 and was independent of the activation of IRF-3. N-terminal lysine residues were shown to play a critical role in the Vif- and Vpr-mediated degradation of IRF-3. These data implicate Vif and Vpr in the disruption of the initial antiviral response and point to the need of HIV-1 to circumvent the antiviral response during the very early phase of replication

  12. A Role of Protein Degradation in Memory Consolidation after Initial Learning and Extinction Learning in the Honeybee ("Apis mellifera")

    Science.gov (United States)

    Felsenberg, Johannes; Dombrowski, Vincent; Eisenhardt, Dorothea

    2012-01-01

    Protein degradation is known to affect memory formation after extinction learning. We demonstrate here that an inhibitor of protein degradation, MG132, interferes with memory formation after extinction learning in a classical appetitive conditioning paradigm. In addition, we find an enhancement of memory formation when the same inhibitor is…

  13. Effect of dietary starch level and high rumen-undegradable protein on endocrine-metabolic status, milk yield, and milk composition in dairy cows during early and late lactation.

    Science.gov (United States)

    Piccioli-Cappelli, F; Loor, J J; Seal, C J; Minuti, A; Trevisi, E

    2014-12-01

    Diet composition defines the amount and type of nutrients absorbed by dairy cows. Endocrine-metabolic interactions can influence these parameters, and so nutrient availability for the mammary gland can significantly vary and affect milk yield and its composition. Six dairy cows in early and then late lactation received, for 28 d in a changeover design, 2 diets designed to provide, within the same stage of lactation, similar amounts of rumen fermentable material but either high starch plus sugar (HS) content or low starch plus sugar content (LS). All diets had similar dietary crude protein and calculated supply of essential amino acids. Dry matter intake within each stage of lactation was similar between groups. Milk yield was similar between groups in early lactation, whereas a higher milk yield was observed in late lactation when feeding HS. At the metabolic level, the main difference observed between the diets in both stages of lactation was lower blood glucose in cows fed LS. The lower glucose availability during consumption of LS caused substantial modifications in the circulating and postprandial pattern of metabolic hormones. Feeding LS versus HS resulted in an increase in the ratio of bovine somatotropin to insulin. This increased mobilization of lipid reserves resulted in higher blood concentrations of nonesterified fatty acids and β-hydroxybutyrate, which contributed to the higher milk fat content in both stages of lactation in the LS group. This greater recourse to body fat stores was confirmed by the greater loss of body weight during early lactation and the slower recovery of body weight in late lactation in cows fed LS. The lower insulin to glucagon ratio observed in cows fed LS in early and late lactation likely caused an increase in hepatic uptake and catabolism of amino acids, as confirmed by the higher blood urea concentrations. Despite the higher catabolism of amino acids in LS in early lactation, similar milk protein output was observed for both

  14. Alfalfa stem tissues: Cell wall deposition, composition, and degradability

    NARCIS (Netherlands)

    Jung, H.G.; Engels, F.M.

    2002-01-01

    Declining cell wall degradability of alfalfa (Medicago sativa L.) stems with maturation limits the nutritional value of alfalfa for ruminants. This study characterized changes in cell wall concentration, composition, and degradability by rumen microbes resulting from alfalfa stem tissue

  15. Degradation of the disease-associated prion protein by a serine protease from lichens

    Science.gov (United States)

    Johnson, C.J.; Bennett, J.P.; Biro, S.M.; Duque-Velasquez, J.C.; Rodriguez, C.M.; Bessen, R.A.; Rocke, T.E.; Bartz, Jason C.

    2011-01-01

    The disease-associated prion protein (PrP(TSE)), the probable etiological agent of the transmissible spongiform encephalopathies (TSEs), is resistant to degradation and can persist in the environment. Lichens, mutualistic symbioses containing fungi, algae, bacteria and occasionally cyanobacteria, are ubiquitous in the environment and have evolved unique biological activities allowing their survival in challenging ecological niches. We investigated PrP(TSE) inactivation by lichens and found acetone extracts of three lichen species (Parmelia sulcata, Cladonia rangiferina and Lobaria pulmonaria) have the ability to degrade prion protein (PrP) from TSE-infected hamsters, mice and deer. Immunoblots measuring PrP levels and protein misfolding cyclic amplification indicated at least two logs of reductions in PrP(TSE). Degradative activity was not found in closely related lichen species or in algae or a cyanobacterium that inhabit lichens. Degradation was blocked by Pefabloc SC, a serine protease inhibitor, but not inhibitors of other proteases or enzymes. Additionally, we found that PrP levels in PrP(TSE)-enriched preps or infected brain homogenates are also reduced following exposure to freshly-collected P. sulcata or an aqueous extract of the lichen. Our findings indicate that these lichen extracts efficiently degrade PrP(TSE) and suggest that some lichens could have potential to inactivate TSE infectivity on the landscape or be a source for agents to degrade prions. Further work to clone and characterize the protease, assess its effect on TSE infectivity and determine which organism or organisms present in lichens produce or influence the protease activity is warranted.

  16. HUWE1 and TRIP12 Collaborate in Degradation of Ubiquitin-Fusion Proteins and Misframed Ubiquitin

    DEFF Research Database (Denmark)

    Poulsen, Esben G; Steinhauer, Cornelia; Lees, Michael

    2012-01-01

    In eukaryotic cells an uncleavable ubiquitin moiety conjugated to the N-terminus of a protein signals the degradation of the fusion protein via the proteasome-dependent ubiquitin fusion degradation (UFD) pathway. In yeast the molecular mechanism of the UFD pathway has been well characterized...... in degradation of the UFD substrate Ub(G76V)-YFP. The most significant hits from the screen were the E3 ubiquitin-protein ligase HUWE1, as well as PSMD7 and PSMD14 that encode proteasome subunits. Accordingly, knock down of HUWE1 led to an increase in the steady state level and a retarded degradation of the UFD...... substrate. Knock down of HUWE1 also led to a stabilization of the physiological UFD substrate UBB(+1). Precipitation experiments revealed that HUWE1 is associated with both the Ub(G76V)-YFP substrate and the 26S proteasome, indicating that it functions late in the UFD pathway. Double knock down of HUWE1...

  17. Application of green fluorescent protein for monitoring phenol-degrading strains

    Directory of Open Access Journals (Sweden)

    Ana Milena Valderrama F.

    2001-07-01

    Full Text Available Several methods have been developed for detecting microorganisms in environmental samples. Some systems for incorporating reporter genes, such as lux or the green fluorescent protein (GFP gene, have been developed recently This study describes gfp gene marking of a phenol degrading strain, its evaluation and monitoring in a bioreactor containing refinery sour water. Tagged strains were obtained having the same physiological and metabolic characteristics as the parent strain. Fluorescent expression was kept stable with no selection for more than 50 consecutive generations and tagged strains were recovered from the bioreactor after forty-five days of phenol-degradation treatment.

  18. Unfolded protein response and activated degradative pathways regulation in GNE myopathy.

    Directory of Open Access Journals (Sweden)

    Honghao Li

    Full Text Available Although intracellular beta amyloid (Aβ accumulation is known as an early upstream event in the degenerative course of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE myopathy, the process by which Aβdeposits initiate various degradative pathways, and their relationship have not been fully clarified. We studied the possible secondary responses after amyloid beta precursor protein (AβPP deposition including unfolded protein response (UPR, ubiquitin proteasome system (UPS activation and its correlation with autophagy system. Eight GNE myopathy patients and five individuals with normal muscle morphology were included in this study. We performed immunofluorescence and immunoblotting to investigate the expression of AβPP, phosphorylated tau (p-tau and endoplasmic reticulum molecular chaperones. Proteasome activities were measured by cleavage of fluorogenic substrates. The expression of proteasome subunits and linkers between proteasomal and autophagy systems were also evaluated by immunoblotting and relative quantitative real-time RT-PCR. Four molecular chaperones, glucose-regulated protein 94 (GRP94, glucose-regulated protein 78 (GRP78, calreticulin and calnexin and valosin containing protein (VCP were highly expressed in GNE myopathy. 20S proteasome subunits, three main proteasome proteolytic activities, and the factors linking UPS and autophagy system were also increased. Our study suggests that AβPP deposition results in endoplasmic reticulum stress (ERS and highly expressed VCP deliver unfolded proteins from endoplasmic reticulum to proteosomal system which is activated in endoplasmic reticulum associated degradation (ERAD in GNE myopathy. Excessive ubiquitinated unfolded proteins are exported by proteins that connect UPS and autophagy to autophagy system, which is activated as an alternative pathway for degradation.

  19. Degradation of Human PDZ-Proteins by Human Alphapapillomaviruses Represents an Evolutionary Adaptation to a Novel Cellular Niche.

    Science.gov (United States)

    Van Doorslaer, Koenraad; DeSalle, Rob; Einstein, Mark H; Burk, Robert D

    2015-06-01

    In order to complete their life cycle, papillomaviruses have evolved to manipulate a plethora of cellular pathways. The products of the human Alphapapillomavirus E6 proteins specifically interact with and target PDZ containing proteins for degradation. This viral phenotype has been suggested to play a role in viral oncogenesis. To analyze the association of HPV E6 mediated PDZ-protein degradation with cervical oncogenesis, a high-throughput cell culture assay was developed. Degradation of an epitope tagged human MAGI1 isoform was visualized by immunoblot. The correlation between HPV E6-induced degradation of hMAGI1 and epidemiologically determined HPV oncogenicity was evaluated using a Bayesian approach within a phylogenetic context. All tested oncogenic types degraded the PDZ-containing protein hMAGI1d; however, E6 proteins isolated from several related albeit non-oncogenic viral types were equally efficient at degrading hMAGI1. The relationship between both traits (oncogenicity and PDZ degradation potential) is best explained by a model in which the potential to degrade PDZ proteins was acquired prior to the oncogenic phenotype. This analysis provides evidence that the ancestor of both oncogenic and non-oncogenic HPVs acquired the potential to degrade human PDZ-containing proteins. This suggests that HPV E6 directed degradation of PDZ-proteins represents an ancient ecological niche adaptation. Phylogenetic modeling indicates that this phenotype is not specifically correlated with oncogenic risk, but may act as an enabling phenotype. The role of PDZ protein degradation in HPV fitness and oncogenesis needs to be interpreted in the context of Alphapapillomavirus evolution.

  20. Degradation of Human PDZ-Proteins by Human Alphapapillomaviruses Represents an Evolutionary Adaptation to a Novel Cellular Niche

    Science.gov (United States)

    Van Doorslaer, Koenraad; DeSalle, Rob; Einstein, Mark H.; Burk, Robert D.

    2015-01-01

    In order to complete their life cycle, papillomaviruses have evolved to manipulate a plethora of cellular pathways. The products of the human Alphapapillomavirus E6 proteins specifically interact with and target PDZ containing proteins for degradation. This viral phenotype has been suggested to play a role in viral oncogenesis. To analyze the association of HPV E6 mediated PDZ-protein degradation with cervical oncogenesis, a high-throughput cell culture assay was developed. Degradation of an epitope tagged human MAGI1 isoform was visualized by immunoblot. The correlation between HPV E6-induced degradation of hMAGI1 and epidemiologically determined HPV oncogenicity was evaluated using a Bayesian approach within a phylogenetic context. All tested oncogenic types degraded the PDZ-containing protein hMAGI1d; however, E6 proteins isolated from several related albeit non-oncogenic viral types were equally efficient at degrading hMAGI1. The relationship between both traits (oncogenicity and PDZ degradation potential) is best explained by a model in which the potential to degrade PDZ proteins was acquired prior to the oncogenic phenotype. This analysis provides evidence that the ancestor of both oncogenic and non-oncogenic HPVs acquired the potential to degrade human PDZ-containing proteins. This suggests that HPV E6 directed degradation of PDZ-proteins represents an ancient ecological niche adaptation. Phylogenetic modeling indicates that this phenotype is not specifically correlated with oncogenic risk, but may act as an enabling phenotype. The role of PDZ protein degradation in HPV fitness and oncogenesis needs to be interpreted in the context of Alphapapillomavirus evolution. PMID:26086730

  1. Degradation of Human PDZ-Proteins by Human Alphapapillomaviruses Represents an Evolutionary Adaptation to a Novel Cellular Niche.

    Directory of Open Access Journals (Sweden)

    Koenraad Van Doorslaer

    2015-06-01

    Full Text Available In order to complete their life cycle, papillomaviruses have evolved to manipulate a plethora of cellular pathways. The products of the human Alphapapillomavirus E6 proteins specifically interact with and target PDZ containing proteins for degradation. This viral phenotype has been suggested to play a role in viral oncogenesis. To analyze the association of HPV E6 mediated PDZ-protein degradation with cervical oncogenesis, a high-throughput cell culture assay was developed. Degradation of an epitope tagged human MAGI1 isoform was visualized by immunoblot. The correlation between HPV E6-induced degradation of hMAGI1 and epidemiologically determined HPV oncogenicity was evaluated using a Bayesian approach within a phylogenetic context. All tested oncogenic types degraded the PDZ-containing protein hMAGI1d; however, E6 proteins isolated from several related albeit non-oncogenic viral types were equally efficient at degrading hMAGI1. The relationship between both traits (oncogenicity and PDZ degradation potential is best explained by a model in which the potential to degrade PDZ proteins was acquired prior to the oncogenic phenotype. This analysis provides evidence that the ancestor of both oncogenic and non-oncogenic HPVs acquired the potential to degrade human PDZ-containing proteins. This suggests that HPV E6 directed degradation of PDZ-proteins represents an ancient ecological niche adaptation. Phylogenetic modeling indicates that this phenotype is not specifically correlated with oncogenic risk, but may act as an enabling phenotype. The role of PDZ protein degradation in HPV fitness and oncogenesis needs to be interpreted in the context of Alphapapillomavirus evolution.

  2. Phosphorylation-dependent signaling controls degradation of DNA mismatch repair protein PMS2.

    Science.gov (United States)

    Hinrichsen, Inga; Weßbecher, Isabel M; Huhn, Meik; Passmann, Sandra; Zeuzem, Stefan; Plotz, Guido; Biondi, Ricardo M; Brieger, Angela

    2017-12-01

    MutLα, a heterodimer consisting of MLH1 and PMS2, plays an important role in DNA mismatch repair and has been shown to be additionally involved in several other important cellular mechanisms. Previous work indicated that AKT could modulate PMS2 stability by phosphorylation. Still, the mechanisms of regulation of MutLα remain unclear. The stability of MutLα subunits was investigated by transiently overexpression of wild type and mutant forms of MLH1 and PMS2 using immunoblotting for measuring the protein levels after treatment. We found that treatment with the cell-permeable serine/threonine phosphatase inhibitor, Calyculin, leads to degradation of PMS2 when MLH1 or its C-terminal domain is missing or if amino acids of MLH1 essential for PMS2 interaction are mutated. In addition, we discovered that the C-terminal tail of PMS2 is relevant for this Calyculin-dependent degradation. A direct involvement of AKT, which was previously described to be responsible for PMS2 degradation, could not be detected. The multi-kinase inhibitor Sorafenib, in contrast, was able to avoid the degradation of PMS2 which postulates that cellular phosphorylation is involved in this process. Together, we show that pharmacologically induced phosphorylation by Calyculin can induce the selective proteasome-dependent degradation of PMS2 but not of MLH1 and that the PMS2 degradation could be blocked by Sorafenib treatment. Curiously, the C-terminal Lynch Syndrome-variants MLH1 L749P and MLH1 Y750X make PMS2 prone to Calyculin induced degradation. Therefore, we conclude that the specific degradation of PMS2 may represent a new mechanism to regulate MutLα. © 2017 Wiley Periodicals, Inc.

  3. Generation of Oxygen Free Radicals by Proflavine: Implication in Protein Degradation

    Directory of Open Access Journals (Sweden)

    Mansour K.M. Gatasheh

    2017-07-01

    Full Text Available Proflavine, an acridine dye, is a known DNA intercalating agent. In the present study, we show that proflavine alone on photoillumination can generate reactive oxygen species (ROS. These proflavine-derived ROS cause damage to proteins, and this effect is enhanced when the divalent metal ion Cu (II is included in the reaction. Bathocuproine, a specific Cu (I sequestering agent, when present in the reaction mixture containing Cu (II, was found to inhibit the protein degradation, showing that Cu (I is an essential intermediate in the reaction. The effect of several scavengers of ROS such as superoxide dismutase, sodium azide, potassium iodide, and thiourea were examined on the protein damaging reaction. Potassium iodide was found to be the most effective in inhibiting protein damage followed by sodium azide and thiourea. Our results indicate the involvement of superoxide, singlet oxygen, triplet oxygen, and hydroxyl radicals in proflavine-induced damage to proteins.

  4. Measurement of local cerebral protein synthesis in vivo: influence of recycling of amino acids derived from protein degradation

    International Nuclear Information System (INIS)

    Smith, C.B.; Deibler, G.E.; Eng, N.; Schmidt, K.; Sokoloff, L.

    1988-01-01

    A quantitative autoradiographic method for the determination of local rates of protein synthesis in brain in vivo is being developed. The method employs L-[1- 14 C]leucine as the radiolabeled tracer. A comprehensive model has been designed that takes into account intracellular and extracellular spaces, intracellular compartmentation of leucine, and the possibility of recycling of unlabeled leucine derived from steady-state degradation of protein into the precursor pool for protein synthesis. We have evaluated the degree of recycling by measuring the ratio of the steady-state precursor pool distribution space for labeled leucine to that of unlabeled leucine. The values obtained were 0.58 in whole brain and 0.47 in liver. These results indicate that there is significant recycling of unlabeled amino acids derived from steady-state protein degradation in both tissues. Any method for the determination of rates of cerebral protein synthesis in vivo with labeled tracers that depends on estimation of precursor pool specific activity in tissue from measurements in plasma must take this recycling into account

  5. A STRUCTURAL OVERVIEW OF GH61 PROTEINS – FUNGAL CELLULOSE DEGRADING POLYSACCHARIDE MONOOXYGENASES

    Directory of Open Access Journals (Sweden)

    Leila Lo Leggio

    2012-09-01

    Full Text Available Recent years have witnessed a spurt of activities in the elucidation of the molecular function of a class of proteins with great potential in biomass degradation. GH61 proteins are of fungal origin and were originally classified in family 61 of the glycoside hydrolases. From the beginning they were strongly suspected to be involved in cellulose degradation because of their expression profiles, despite very low detectable endoglucanase activities. A major breakthrough came from structure determination of the first members, establishing the presence of a divalent metal binding site and a similarity to bacterial proteins involved in chitin degradation. A second breakthrough came from the identification of cellulase boosting activity dependent on the integrity of the metal binding site. Finally very recently GH61 proteins were demonstrated to oxidatively cleave crystalline cellulose in a Cu and reductant dependant manner. This mini-review in particular focuses on the contribution that structure elucidation has made in the understanding of GH61 molecular function and reviews the currently known structures and the challenges remaining ahead for exploiting this new class of enzymes to the full.

  6. A structural overview of GH61 proteins – fungal cellulose degrading polysaccharide monooxygenases

    Directory of Open Access Journals (Sweden)

    Leila Lo Leggio

    2012-09-01

    Full Text Available Recent years have witnessed a spurt of activities in the elucidation of the molecular function of a class of proteins with great potential in biomass degradation. GH61 proteins are of fungal origin and were originally classified in family 61 of the glycoside hydrolases. From the beginning they were strongly suspected to be involved in cellulose degradation because of their expression profiles, despite very low detectable endoglucanase activities. A major breakthrough came from structure determination of the first members, establishing the presence of a divalent metal binding site and a similarity to bacterial proteins involved in chitin degradation. A second breakthrough came from the identification of cellulase boosting activity dependent on the integrity of the metal binding site. Finally very recently GH61 proteins were demonstrated to oxidatively cleave crystalline cellulose in a Cu and reductant dependant manner. This mini-review in particular focuses on the contribution that structure elucidation has made in the understanding of GH61 molecular function and reviews the currently known structures and the challenges remaining ahead for exploiting this new class of enzymes to the full.

  7. HUWE1 and TRIP12 collaborate in degradation of ubiquitin-fusion proteins and misframed ubiquitin.

    Directory of Open Access Journals (Sweden)

    Esben G Poulsen

    Full Text Available In eukaryotic cells an uncleavable ubiquitin moiety conjugated to the N-terminus of a protein signals the degradation of the fusion protein via the proteasome-dependent ubiquitin fusion degradation (UFD pathway. In yeast the molecular mechanism of the UFD pathway has been well characterized. Recently the human E3 ubiquitin-protein ligase TRIP12 was connected with the UFD pathway, but little is otherwise known about this system in mammalian cells. In the present work, we utilized high-throughput imaging on cells transfected with a targeted siRNA library to identify components involved in degradation of the UFD substrate Ub(G76V-YFP. The most significant hits from the screen were the E3 ubiquitin-protein ligase HUWE1, as well as PSMD7 and PSMD14 that encode proteasome subunits. Accordingly, knock down of HUWE1 led to an increase in the steady state level and a retarded degradation of the UFD substrate. Knock down of HUWE1 also led to a stabilization of the physiological UFD substrate UBB(+1. Precipitation experiments revealed that HUWE1 is associated with both the Ub(G76V-YFP substrate and the 26S proteasome, indicating that it functions late in the UFD pathway. Double knock down of HUWE1 and TRIP12 resulted in an additive stabilization of the substrate, suggesting that HUWE1 and TRIP12 function in parallel during UFD. However, even when both HUWE1 and TRIP12 are downregulated, ubiquitylation of the UFD substrate was still apparent, revealing functional redundancy between HUWE1, TRIP12 and yet other ubiquitin-protein ligases.

  8. Protection of a protein against irradiation-induced degradation by additives in the solid state

    International Nuclear Information System (INIS)

    Shalaev, E.; Reddy, R.; Kimball, R.N.; Weinschenk, M.F.; Guinn, M.; Margulis, L.

    2003-01-01

    The impact of ionizing radiation on a globular protein (porcine somatotropin, pST) in the solid state was studied using rate of dissolution, high-performance liquid chromatography, and Electron spin resonance (ESR) in the presence of different additives. o-Vanillin stabilized pST against irradiation-induced degradation whereas effects of trolox and isopropyl alcohol were less significant. Stabilization effect of o-vanillin has been related to the energy transfer from pST molecules to the additive which was facilitated by formation of covalent bonds between o-vanillin and pST molecules. Anticorrelation between the level of free radicals and chemical degradation (i.e. degradation increased with decrease in a free radical level) was observed in the presence of o-vanillin

  9. Precision-feeding dairy heifers a high rumen-undegradable protein diet with different proportions of dietary fiber and forage-to-concentrate ratios.

    Science.gov (United States)

    Koch, L E; Gomez, N A; Bowyer, A; Lascano, G J

    2017-12-01

    The addition of dietary fiber can alter nutrient and N utilization in precision-fed dairy heifers and may further benefit from higher inclusion levels of RUP. The objective of this experiment was to determine the effects of feeding a high-RUP diet when dietary fiber content was manipulated within differing forage-to-concentrate ratios (F:C) on nutrient utilization of precision-fed dairy heifers. Six rumen-cannulated Holstein heifers (555.4 ± 31.4 kg BW; 17.4 ± 0.1 mo) were randomly assigned to 2 levels of forage, high forage (HF; 60% forage) or low forage (LF; 45% forage), and to a fiber proportion sequence (low fiber: 100% oat hay and silage [OA], 0% wheat straw [WS]; medium fiber: 83.4% OA, 16.6% WS; and high fiber: 66.7% OA, 33.3% WS) administered according to a split-plot 3 × 3 Latin square design (21-d periods). Similar levels of N intake (1.70 g N/kg BW) and RUP (55% of CP) were provided. Data were analyzed as a split-plot, 3 × 3 Latin square design using a mixed model with fixed effects of period and treatment. A repeated measures model was used with data that had multiple measurements over time. No differences were observed for DM, OM, NDF, or ADF apparent digestibility coefficients (dC) between HF- and LF-fed heifers. Heifers receiving LF diets had greater starch dC compared to HF heifers. Increasing the fiber level through WS addition resulted in a linear reduction of OM dC. There was a linear interaction for DM dC with a concurrent linear interaction in NDF dC. Nitrogen intake, dC, and retention did not differ; however, urine and total N excretion increased linearly with added fiber. Predicted microbial CP flow (MP) linearly decreased with WS inclusion mainly in LF heifers, as indicated by a significant interaction between F:C and WS. Rumen pH linearly increased with WS addition, although no F:C effect was detected. Ruminal ammonia concentration had an opposite linear effect with respect to MP as WS increased. Diets with the higher proportion of

  10. Gold nanoparticles enhance the X-ray-induced degradation of human centrin 2 protein

    Energy Technology Data Exchange (ETDEWEB)

    Brun, Emilie [Laboratoire de Chimie Physique, CNRS UMR 8000, Universite Paris-Sud 11, Bat. 350, 91405 Orsay Cedex (France); Duchambon, Patricia; Blouquit, Yves [INSERM U759, Imagerie Integrative, Campus Universitaire d' Orsay, Bat. 112, Institut Curie, Centre de Recherche, Laboratoire R. Latarjet, Campus Universitaire d' Orsay, 91405 Orsay Cedex (France); Keller, Gerard [UMR CNRS 8612, Physico-Chimie-Pharmacotechnie-Biopharmacie, Universite Paris 11, Faculte de Pharmacie, 5 rue Jean-Baptiste Clement, 92296 Chatenay-Malabry (France); Sanche, Leon [Groupe en Sciences des Radiations, Departement de Medecine Nucleaire et Radiobiologie, Faculte de Medecine, Universite de Sherbrooke, Sherbrooke, Quebec, Canada J1H 5N4 (Canada); Sicard-Roselli, Cecile [Laboratoire de Chimie Physique, CNRS UMR 8000, Universite Paris-Sud 11, Bat. 350, 91405 Orsay Cedex (France)], E-mail: cecile.sicard@u-psud.fr

    2009-03-15

    In the war against cancer, radiotherapy is a prominent tool but counterbalanced by the fact that it also induces damages in healthy tissues. Nanotechnologies could open a new possibility to decrease these side effects. In particular, gold nanoparticles (GNPs) could be used as radio-sensitizers. As the role of proteins in the processes leading to cell death cannot be neglected, their radio-sensitization by GNPs is of great interest. This is particularly true in the case of the human centrin 2 protein, which has been proposed to be involved in DNA repair processes. To investigate this effect, we quantified for the first time the degradation of this protein in a gold colloidal solution when submitted to X-rays. We showed that the X-ray-induced degradation of the human centrin 2 protein is enhanced 1.5-fold in the presence of GNPs, even though no covalent bond exists between protein and GNPs. Among the conditions tested, the maximum enhancement was found with the higher GNP:protein ratio of 2x10{sup -4} and with the higher X-ray energy of 49 keV.

  11. Gold nanoparticles enhance the X-ray-induced degradation of human centrin 2 protein

    International Nuclear Information System (INIS)

    Brun, Emilie; Duchambon, Patricia; Blouquit, Yves; Keller, Gerard; Sanche, Leon; Sicard-Roselli, Cecile

    2009-01-01

    In the war against cancer, radiotherapy is a prominent tool but counterbalanced by the fact that it also induces damages in healthy tissues. Nanotechnologies could open a new possibility to decrease these side effects. In particular, gold nanoparticles (GNPs) could be used as radio-sensitizers. As the role of proteins in the processes leading to cell death cannot be neglected, their radio-sensitization by GNPs is of great interest. This is particularly true in the case of the human centrin 2 protein, which has been proposed to be involved in DNA repair processes. To investigate this effect, we quantified for the first time the degradation of this protein in a gold colloidal solution when submitted to X-rays. We showed that the X-ray-induced degradation of the human centrin 2 protein is enhanced 1.5-fold in the presence of GNPs, even though no covalent bond exists between protein and GNPs. Among the conditions tested, the maximum enhancement was found with the higher GNP:protein ratio of 2x10 -4 and with the higher X-ray energy of 49 keV

  12. Efficacy of different essential oils in modulating rumen fermentation in vitro using buffalo rumen liquor

    Directory of Open Access Journals (Sweden)

    Debashis Roy

    2014-04-01

    Full Text Available Aim: Present study was conducted to examine the modulatory effect of different essential oils on rumen fermentation pattern in vitro using wheat straw based diet (concentrate: wheat straw 50:50. Materials and Methods: Four essential oils i.e. cinnamon, garlic, oregano and rosemary oils were tested at concentration of 0, 30, 300 and 600 mg/litre (ppm of total culture fluid using in vitro gas production technique. Total gas production, methane production, nutrient degradability, volatile fatty acid (VFA production and ammonia nitrogen concentration were studied in vitro using buffalo rumen liquor. Results: Results indicated that all four essential oils decreased gas production significantly (P<0.05 at 600ppm concentration. However, in case of garlic oil, 300 ppm concentration was also found to be effective in decreasing total gas production. Reduction in methane production was found maximum (P<0.05 at higher doses in most of the oils. Maximum reduction in methane was noticed with garlic oil at 600ppm dose. Ammonia-N concentration was also decreased significantly (P<0.05 with essential oils and was found minimum with oregano oil at 600 ppm dose. Partition factor was found to be significantly (P<0.05 higher in 600 ppm concentration of garlic and oregano oil. The degradability of dry matter decreased significantly with higher concentration of essential oil in most of treatment combinations. Conclusion: Supplementation with different essential oils on wheat straw based diet modulates rumen fermentation and reduced methane and ammonia- N production and improved utilization of nutrients.

  13. Dma1-dependent degradation of SIN proteins during meiosis in Schizosaccharomyces pombe.

    Science.gov (United States)

    Krapp, Andrea; Simanis, Viesturs

    2014-07-15

    The Schizosaccharomyces pombe septation initiation network (SIN) is required for cytokinesis during vegetative growth and for spore formation during meiosis. Regulation of the SIN during mitosis has been studied extensively, but less is known about its meiotic regulation. Here, we show that several aspects of SIN regulation differ between mitosis and meiosis. First, the presence of GTP-bound Spg1p is not the main determinant of the timing of Cdc7p and Sid1p association with the spindle pole body (SPB) during meiosis. Second, the localisation dependencies of SIN proteins differ from those in mitotic cells, suggesting a modified functional organisation of the SIN during meiosis. Third, there is stage-specific degradation of SIN components in meiosis; Byr4p is degraded after meiosis I, whereas the degradation of Cdc7p, Cdc11p and Sid4p occurs after the second meiotic division and depends upon the ubiquitin ligase Dma1p. Finally, Dma1p-dependent degradation is not restricted to the SIN, as we show that Dma1p is needed for the degradation of Mcp6p (also known as Hrs1p) during meiosis I. Taken together, these data suggest that stage-specific targeted proteolysis plays an important role in regulating meiotic progression. © 2014. Published by The Company of Biologists Ltd.

  14. Protein oxidation and degradation during proliferative senescence of human MRC-5 fibroblasts.

    Science.gov (United States)

    Sitte, N; Merker, K; von Zglinicki, T; Grune, T

    2000-03-01

    One of the highlights of age-related changes of cellular metabolism is the accumulation of oxidized proteins. The aging process on a cellular level can be treated either as the ongoing proliferation until a certain number of cell divisions is reached (the Hayflick limit) or as the aging of nondividing cells, that is, the age-related changes in cells without proliferation. The present investigation was undertaken to reveal the changes in protein turnover, proteasome activity, and protein oxidation status during proliferative senescence. We were able to demonstrate that the activity of the cytosolic proteasomal system declines dramatically during the proliferative senescence of human MRC-5 fibroblasts. Regardless of the loss in activity, it could be demonstrated that there are no changes in the transcription and translation of proteasomal subunits. This decline in proteasome activity was accompanied by an increased concentration of oxidized proteins. Cells at higher proliferation stages were no longer able to respond with increased degradation of endogenous [(35)S]-Met-radiolabeled proteins after hydrogen peroxide- or quinone-induced oxidative stress. It could be demonstrated that oxidized proteins in senescent human MRC-5 fibroblasts are not as quickly removed as they are in young cells. Therefore, our study demonstrates that the accumulation of oxidized proteins and decline in protein turnover and activity of the proteasomal system are not only a process of postmitotic aging but also occur during proliferative senescence and result in an increased half-life of oxidized proteins.

  15. New strategy for renal fibrosis: Targeting Smad3 proteins for ubiquitination and degradation.

    Science.gov (United States)

    Wang, Xin; Feng, Shaozhen; Fan, Jinjin; Li, Xiaoyan; Wen, Qiong; Luo, Ning

    2016-09-15

    Smad3 is a critical signaling protein in renal fibrosis. Proteolysis targeting chimeric molecules (PROTACs) are small molecules designed to degrade target proteins via ubiquitination. They have three components: (1) a recognition motif for E3 ligase; (2) a linker; and (3) a ligand for the target protein. We aimed to design a new PROTAC to prevent renal fibrosis by targeting Smad3 proteins and using hydroxylated pentapeptide of hypoxia-inducible factor-1α as the recognition motif for von Hippel-Lindau (VHL) ubiquitin ligase (E3). Computer-aided drug design was used to find a specific ligand targeting Smad3. Surface plasmon resonance (SPR) was used to verify and optimize screening results. Synthesized PROTAC was validated by two-stage mass spectrometry. The PROTAC's specificity for VHL (E3 ligase) was proved with two human renal carcinoma cell lines, 786-0 (VHL(-)) and ACHN (VHL(+)), and its anti-fibrosis effect was tested in renal fibrosis cell models. Thirteen small molecular compounds (SMCs) were obtained from the Enamine library using GLIDE molecular docking program. SPR results showed that #8 SMC (EN300-72284) combined best with Smad3 (KD=4.547×10(-5)M). Mass spectrometry showed that synthesized PROTAC had the correct peptide molecular weights. Western blot showed Smad3 was degraded by PROTAC with whole-cell lysate of ACHN but not 786-0. Degradation, but not ubiquitination, of Smad3 was inhibited by proteasome inhibitor MG132. The upregulation of fibronectin and Collagen I induced by TGF-β1 in both renal fibroblast and mesangial cells were inhibited by PROTAC. The new PROTAC might prevent renal fibrosis by targeting Smad3 for ubiquitination and degradation. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Rumen microbial communities influence metabolic phenotypes in lambs

    Directory of Open Access Journals (Sweden)

    Diego P. Morgavi

    2015-10-01

    Full Text Available The rumen microbiota is an essential part of ruminants forging their nutrition and health. Despite its importance, it is not fully understood how various groups of rumen microbes affect host-microbe relationships and functions. The aim of the study was to simultaneously explore the rumen microbiota and the metabolic phenotype of lambs for identifying host-microbe associations and potential biomarkers of digestive functions. Twin lambs, separated in two groups after birth were exposed to practices (isolation and gavage with rumen fluid with protozoa or protozoa-depleted that differentially restricted the acquisition of microbes. Rumen microbiota, fermentation parameters, digestibility and growth were monitored for up to 31 weeks of age. Microbiota assembled in isolation from other ruminants lacked protozoa and had low bacterial and archaeal diversity whereas digestibility was not affected. Exposure to adult sheep microbiota increased bacterial and archaeal diversity independently of protozoa presence. For archaea, Methanomassiliicoccales displaced Methanosphaera. Notwithstanding, protozoa induced differences in functional traits such as digestibility and significantly shaped bacterial community structure, notably Ruminococcaceae and Lachnospiraceae lower up to 6 folds, Prevotellaceae lower by ~40%, and Clostridiaceae and Veillonellaceae higher up to 10 folds compared to microbiota without protozoa. An orthogonal partial least squares-discriminant analysis of urinary metabolome matched differences in microbiota structure. Discriminant metabolites were mainly involved in amino acids and protein metabolic pathways while a negative interaction was observed between methylotrophic methanogens Methanomassiliicoccales and trimethylamine N-oxide. These results stress the influence of gut microbes on animal phenotype and show the potential of metabolomics for monitoring rumen microbial functions.

  17. Nck adaptor proteins link Tks5 to invadopodia actin regulation and ECM degradation.

    Science.gov (United States)

    Stylli, Stanley S; Stacey, T T I; Verhagen, Anne M; Xu, San San; Pass, Ian; Courtneidge, Sara A; Lock, Peter

    2009-08-01

    Invadopodia are actin-based projections enriched with proteases, which invasive cancer cells use to degrade the extracellular matrix (ECM). The Phox homology (PX)-Src homology (SH)3 domain adaptor protein Tks5 (also known as SH3PXD2A) cooperates with Src tyrosine kinase to promote invadopodia formation but the underlying pathway is not clear. Here we show that Src phosphorylates Tks5 at Y557, inducing it to associate directly with the SH3-SH2 domain adaptor proteins Nck1 and Nck2 in invadopodia. Tks5 mutants unable to bind Nck show reduced matrix degradation-promoting activity and recruit actin to invadopodia inefficiently. Conversely, Src- and Tks5-driven matrix proteolysis and actin assembly in invadopodia are enhanced by Nck1 or Nck2 overexpression and inhibited by Nck1 depletion. We show that clustering at the plasma membrane of the Tks5 inter-SH3 region containing Y557 triggers phosphorylation at this site, facilitating Nck recruitment and F-actin assembly. These results identify a Src-Tks5-Nck pathway in ECM-degrading invadopodia that shows parallels with pathways linking several mammalian and pathogen-derived proteins to local actin regulation.

  18. The Escherichia coli O157:H7 bovine rumen fluid proteome reflects adaptive bacterial responses

    OpenAIRE

    Kudva, Indira T; Stanton, Thaddeus B; Lippolis, John D

    2014-01-01

    Background To obtain insights into Escherichia coli O157:H7 (O157) survival mechanisms in the bovine rumen, we defined the growth characteristics and proteome of O157 cultured in rumen fluid (RF; pH 6.0-7.2 and low volatile fatty acid content) obtained from rumen-fistulated cattle fed low protein content “maintenance diet” under diverse in vitro conditions. Results Bottom-up proteomics (LC-MS/MS) of whole cell-lysates of O157 cultured under anaerobic conditions in filter-sterilized RF (fRF; d...

  19. Lysine-Less Variants of Spinal Muscular Atrophy SMN and SMNΔ7 Proteins Are Degraded by the Proteasome Pathway

    Directory of Open Access Journals (Sweden)

    Raúl Sánchez-Lanzas

    2017-12-01

    Full Text Available Spinal muscular atrophy is due to mutations affecting the SMN1 gene coding for the full-length protein (survival motor neuron; SMN and the SMN2 gene that preferentially generates an exon 7-deleted protein (SMNΔ7 by alternative splicing. To study SMN and SMNΔ7 degradation in the cell, we have used tagged versions at the N- (Flag or C-terminus (V5 of both proteins. Transfection of those constructs into HeLa cells and treatment with cycloheximide showed that those protein constructs were degraded. Proteasomal degradation usually requires prior lysine ubiquitylation. Surprisingly, lysine-less variants of both proteins tagged either at N- (Flag or C-terminus (V5 were also degraded. The degradation of the endogenous SMN protein, and the protein constructs mentioned above, was mediated by the proteasome, as it was blocked by lactacystin, a specific and irreversible proteasomal inhibitor. The results obtained allowed us to conclude that SMN and SMNΔ7 proteasomal degradation did not absolutely require internal ubiquitylation nor N-terminal ubiquitylation (prevented by N-terminal tagging. While the above conclusions are firmly supported by the experimental data presented, we discuss and justify the need of deep proteomic techniques for the study of SMN complex components (orphan and bound turn-over to understand the physiological relevant mechanisms of degradation of SMN and SMNΔ7 in the cell.

  20. Degradation of brown adipocyte purine nucleotides regulates uncoupling protein 1 activity

    Directory of Open Access Journals (Sweden)

    Tobias Fromme

    2018-02-01

    Full Text Available Objective: Non-shivering thermogenesis in mammalian brown adipose tissue depends on thermogenic uncoupling protein 1. Its activity is triggered by free fatty acids while purine nucleotides mediate inhibition. During activation, it is thought that free fatty acids overcome purine-mediated inhibition. We measured the cellular concentration and the release of purine nucleotide metabolites to uncover a possible role of purine nucleotide degradation in uncoupling protein 1 activation. Methods: With mass spectrometry, purine nucleotide metabolites were quantified in cellular homogenates and supernatants of cultured primary brown adipocytes. We also determined oxygen consumption in response to a β-adrenergic agonist. Results: Upon adrenergic activation, brown adipocytes decreased the intracellular concentration of inhibitory nucleotides (ATP, ADP, GTP and GDP and released the respective degradation products. At the same time, an increase in cellular calcium occurred. None of these phenomena occurred in white adipocytes or myotubes. The brown adipocyte expression of enzymes implicated in purine metabolic remodeling is altered upon cold exposure. Pharmacological and genetic interference of purine metabolism altered uncoupling protein 1 mediated uncoupled respiration. Conclusion: Adrenergic stimulation of brown adipocytes lowers the intracellular concentration of purine nucleotides, thereby contributing to uncoupling protein 1 activation. Keywords: Purine nucleotides, Uncoupling protein 1, Brown adipose tissue, Non-shivering thermogenesis, HILIC-MS/MS, Guanosine monophosphate reductase

  1. Methods for the visualization and analysis of extracellular matrix protein structure and degradation.

    Science.gov (United States)

    Leonard, Annemarie K; Loughran, Elizabeth A; Klymenko, Yuliya; Liu, Yueying; Kim, Oleg; Asem, Marwa; McAbee, Kevin; Ravosa, Matthew J; Stack, M Sharon

    2018-01-01

    This chapter highlights methods for visualization and analysis of extracellular matrix (ECM) proteins, with particular emphasis on collagen type I, the most abundant protein in mammals. Protocols described range from advanced imaging of complex in vivo matrices to simple biochemical analysis of individual ECM proteins. The first section of this chapter describes common methods to image ECM components and includes protocols for second harmonic generation, scanning electron microscopy, and several histological methods of ECM localization and degradation analysis, including immunohistochemistry, Trichrome staining, and in situ zymography. The second section of this chapter details both a common transwell invasion assay and a novel live imaging method to investigate cellular behavior with respect to collagen and other ECM proteins of interest. The final section consists of common electrophoresis-based biochemical methods that are used in analysis of ECM proteins. Use of the methods described herein will enable researchers to gain a greater understanding of the role of ECM structure and degradation in development and matrix-related diseases such as cancer and connective tissue disorders. © 2018 Elsevier Inc. All rights reserved.

  2. Modulation of intracellular protein degradation by SSB1-SIS1 chaperon system in yeast S. cerevisiae.

    Science.gov (United States)

    Ohba, M

    1997-06-09

    In prokaryotes, DnaK-DnaJ chaperon is involved in the protein degradation catalyzed by proteases La and ClpA/B complex as shown in E. coli. To extend this into eukaryotic cells, we examined the effects of hsp70 genes, SSA1 and SSB1, and DnaJ genes, SIS1 and YDJ1, on the growth of proteasome subunit mutants of the yeast S. cerevisiae. The results identified SSB1 and SIS1 as a pair of chaperon genes specifically involved in efficient protein turnover in the yeast, whose overexpression suppressed the growth defects caused by the proteasome mutations. Moreover, a single amino acid substitution in the putative peptide-binding site of SSB1 protein profoundly enhanced the suppression activity, indicating that the activity is mediated by the peptide-binding activity of this chaperon. Thus SSB1, with its partner DnaJ, SIS1, modulates the efficiency of protein turnover through its chaperon activity.

  3. Retinoblastoma protein co-purifies with proteasomal insulin-degrading enzyme: Implications for cell proliferation control

    Energy Technology Data Exchange (ETDEWEB)

    Radulescu, Razvan T., E-mail: ratura@gmx.net [Molecular Concepts Research (MCR), Muenster (Germany); Duckworth, William C. [Department of Medicine, Phoenix VA Health Care System, Phoenix, AZ (United States); Levy, Jennifer L. [Research Service, Phoenix VA Health Care System, Phoenix, AZ (United States); Fawcett, Janet, E-mail: janet.fawcett@va.gov [Research Service, Phoenix VA Health Care System, Phoenix, AZ (United States)

    2010-04-30

    Previous investigations on proteasomal preparations containing insulin-degrading enzyme (IDE; EC 3.4.24.56) have invariably yielded a co-purifying protein with a molecular weight of about 110 kDa. We have now found both in MCF-7 breast cancer and HepG2 hepatoma cells that this associated molecule is the retinoblastoma tumor suppressor protein (RB). Interestingly, the amount of RB in this protein complex seemed to be lower in HepG2 vs. MCF-7 cells, indicating a higher (cytoplasmic) protein turnover in the former vs. the latter cells. Moreover, immunofluorescence showed increased nuclear localization of RB in HepG2 vs. MCF-7 cells. Beyond these subtle differences between these distinct tumor cell types, our present study more generally suggests an interplay between RB and IDE within the proteasome that may have important growth-regulatory consequences.

  4. Life and death of proteins after protease cleavage: protein degradation by the N-end rule pathway.

    Science.gov (United States)

    Dissmeyer, Nico; Rivas, Susana; Graciet, Emmanuelle

    2018-05-01

    Contents Summary 929 I. conservation and diversity of N-end rule pathways 929 II. Defensive functions of the N-end rule pathway in plants 930 III. Proteases and degradation by the N-end rule pathway 930 IV. New proteomics approaches for the identification of N-end rule substrates 932 V. Concluding remarks 932 Acknowledgements 934 References 934 SUMMARY: The N-end rule relates the stability of a protein to the identity of its N-terminal residue and some of its modifications. Since its discovery in the 1980s, the repertoire of N-terminal degradation signals has expanded, leading to a diversity of N-end rule pathways. Although some of these newly discovered N-end rule pathways remain largely unexplored in plants, recent discoveries have highlighted roles of N-end rule-mediated protein degradation in plant defense against pathogens and in cell proliferation during organ growth. Despite this progress, a bottleneck remains the proteome-wide identification of N-end rule substrates due to the prerequisite for endoproteolytic cleavage and technical limitations. Here, we discuss the recent diversification of N-end rule pathways and their newly discovered functions in plant defenses, stressing the role of proteases. We expect that novel proteomics techniques (N-terminomics) will be essential for substrate identification. We review these methods, their limitations and future developments. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

  5. Ubiquitination is absolutely required for the degradation of hypoxia-inducible factor - 1 alpha protein in hypoxic conditions

    International Nuclear Information System (INIS)

    Wang, Ronghai; Zhang, Ping; Li, Jinhang; Guan, Hongzai; Shi, Guangjun

    2016-01-01

    The hypoxia-inducible factor (HIF) is recognized as the master regulator of hypoxia response. HIF-α subunits expression are tightly regulated. In this study, our data show that ts20 cells still expressed detectable E1 protein even at 39.5° C for 12 h, and complete depletion of E1 protein expression at 39.5° C by siRNA enhanced HIF-1α and P53 protein expression. Further inhibition of E1 at 39.5 °C by siRNA, or E1 inhibitor Ube1-41 completely blocked HIF-1α degradation. Moreover, immunoprecipitations of co-transfection of HA-ubiquitin and FLAG–HIF–1α plasmids directly confirmed the involvement of ubiquitin in the hypoxic degradation of HIF-1α. Additionally, hypoxic HIF-1 α degradation is independent of HAF, RACK1, sumoylation or nuclear/cytoplasmic localization. Taken together, our data suggest that constitutive HIF-1α protein degradation in hypoxia is absolutely ubiquitination-dependent, and unidentified E3 ligase may exist for this degradation pathway. - Highlights: • HIF-1α protein is constitutively degraded in hypoxic conditions. • Requirement of ubiquitination for HIF-1α degradation in hypoxia. • Hypoxic HIF-1α degradation is independent of HAF, RACK1, sumoylation or nuclear/cytoplasmic localization.

  6. Ubiquitination is absolutely required for the degradation of hypoxia-inducible factor - 1 alpha protein in hypoxic conditions

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Ronghai [Department of Urology, Linzi District People' s Hospital, Zibo, 255400 (China); Zhang, Ping, E-mail: zpskx001@163.com [Department of Gynecology, Qingdao Municipal Hospital, Qingdao, 266011 (China); Li, Jinhang [Department of Gynecology, Qingdao Municipal Hospital, Qingdao, 266011 (China); Guan, Hongzai [Laboratory Department, School of Medicine, Qingdao University, Qingdao, 266071 (China); Shi, Guangjun, E-mail: qdmhshigj@yahoo.com [Department of Hepatobiliary Surgery, Qingdao Municipal Hospital, Qingdao, 266071 (China)

    2016-01-29

    The hypoxia-inducible factor (HIF) is recognized as the master regulator of hypoxia response. HIF-α subunits expression are tightly regulated. In this study, our data show that ts20 cells still expressed detectable E1 protein even at 39.5° C for 12 h, and complete depletion of E1 protein expression at 39.5° C by siRNA enhanced HIF-1α and P53 protein expression. Further inhibition of E1 at 39.5 °C by siRNA, or E1 inhibitor Ube1-41 completely blocked HIF-1α degradation. Moreover, immunoprecipitations of co-transfection of HA-ubiquitin and FLAG–HIF–1α plasmids directly confirmed the involvement of ubiquitin in the hypoxic degradation of HIF-1α. Additionally, hypoxic HIF-1 α degradation is independent of HAF, RACK1, sumoylation or nuclear/cytoplasmic localization. Taken together, our data suggest that constitutive HIF-1α protein degradation in hypoxia is absolutely ubiquitination-dependent, and unidentified E3 ligase may exist for this degradation pathway. - Highlights: • HIF-1α protein is constitutively degraded in hypoxic conditions. • Requirement of ubiquitination for HIF-1α degradation in hypoxia. • Hypoxic HIF-1α degradation is independent of HAF, RACK1, sumoylation or nuclear/cytoplasmic localization.

  7. Studies to Prevent Degradation of Recombinant Fc-Fusion Protein Expressed in Mammalian Cell Line and Protein Characterization

    Directory of Open Access Journals (Sweden)

    Sanjukta Chakrabarti

    2016-06-01

    Full Text Available Clipping of recombinant proteins is a major issue in animal cell cultures. A recombinant Fc-fusion protein, VEGFR1(D1–D3-Fc expressed in CHOK1SV GS-KO cells was observed to be undergoing clippings in lab scale cultures. Partial cleaving of expressed protein initiated early on in cell culture and was observed to increase over time in culture and also on storage. In this study, a few parameters were explored in a bid to inhibit clipping in the fusion protein The effects of culture temperature, duration of culture, the addition of an anti-clumping agent, ferric citrate and use of protease inhibitor cocktail on inhibition of proteolysis of the Fc fusion were studied. Lowering of culture temperature from 37 to 30 °C alone appears to be the best solution for reducing protein degradation from the quality, cost and regulatory points of view. The obtained Fc protein was characterized and found to be in its stable folded state, exhibiting a high affinity for its ligand and also biological and functional activities.

  8. KECERNAAN IN SACCO HIJAUAN LEGUMINOSA DAN HIJAUAN NON- LEGUMINOSA DALAM RUMEN SAPI PERANAKAN ONGOLE

    Directory of Open Access Journals (Sweden)

    Rendi Fathoni Hadi

    2012-06-01

    values of ADF: GL 61.27%; SW 43.95%; TP 56.53%; BR 40.11%; KL 21.08%; NG 44.66%; and CO 69.15%. There were significant differences (P<0.05 on the degradation of DM, OM, CP, NDF, and ADF. It is concluded that not all of legume has higher DT values of DM, OM, CP, NDF, and ADF fraction than non-legume. There is a tendency that the longer retention time in the rumen, the higher degradation rate. (Keywords: Legume, Non-legume, In sacco rumen

  9. Copper-mediated oxidative degradation of catecholamines and oxidative damage of protein

    Energy Technology Data Exchange (ETDEWEB)

    Goncalves, P.R.; Harria, M.I.N.; Felix, J.M.; Hoffmann, M.E. [Universidade Estadual de Campinas, SP (Brazil). Inst. de Biologia

    1997-12-31

    Full text. Degradative oxidation of catecholamines has been a matter of large interest in recent years due to the evidences associating their autoxidation with the etiology of neurotoxic and cardiotoxic processes. In this work we present data on the degradative oxidation of catecholamines of physiological importance: isoproterenol (IP), epinephrine (EP), norepinephrine (NEP), deoxyepinephrine (DEP) and dopamine (DA). The degradative oxidation of the catecholamines was followed by measurement of spectral changes and oxygen consumption by neutral aqueous solutions. The data show that Cu{sup 2+} strongly accelerated the rate of catecholamine oxidation, following the decreasing order; EP>DEP>IP>NEP>DA. The production of superoxide anion radical during catecholamine oxidation was very slow, even in the presence of Cu{sup 2+}. The ability of IP to induce damages on bovine serum albumin (BSA) was determined by measuring the formation of carbonyl-groups in the protein, detected by reduction with tritiated Na BH{sub 4}. The incubation of BSA with IP (50-500{mu}M), in the presence of 100{mu}M Cu{sup 2+} leaded to an increased and dose dependent {sup 3} H-incorporation by the oxidized protein. The production of oxidative damage by IP/Cu{sup 2+} was accompanied by marked BSA fragmentation, detected by SDS-polyacrylamide gel dependent (25-400{mu}M IP) des appearance of the original BSA band and appearance of smaller fragments spread in the gel, when incubation has been done in the presence of 100{mu}M Cu{sup 2+}. These results suggest that copper-catalysed oxidative degradation of proteins induced by catecholamines might be critically involved in the toxic action of these molecules

  10. Feeding value of wheat-based thin stillage: in vitro protein degradability and effects on ruminal fermentation.

    Science.gov (United States)

    Iwanchysko, P; McKinnon, J J; Mustafa, A F; Christensen, D A; McCartney, D

    1999-10-01

    Two experiments were conducted to evaluate the nutritive value of wheat-based thin stillage as a fluid source for ruminants. In vitro CP degradability of thin stillage was estimated relative to canola meal and heated canola meal in a completely randomized design. Four ruminally cannulated steers were used in a double cross-over design to determine the effects of consuming thin stillage or water as drinking sources on ruminal fermentation traits. The in vitro CP degradability of thin stillage (55.4%) was lower (Pstillage was higher (Pstillage than for water-fed steers through most of the collection period. Ruminal fluid and particulate matter passage rates were not affected by treatment and averaged .165 and .06 /h, respectively. The amount of thin stillage and water that did not equilibrate with the ruminal fluid and, thus, was considered to bypass the rumen was estimated to be 51.9 and 59.2% of total fluid consumed, respectively. Feeding wheat-based thin stillage had no adverse effects on ruminal metabolism.

  11. Quantifying protein synthesis and degradation in Arabidopsis by dynamic 13CO2 labeling and analysis of enrichment in individual amino acids in their free pools and in protein.

    Science.gov (United States)

    Ishihara, Hirofumi; Obata, Toshihiro; Sulpice, Ronan; Fernie, Alisdair R; Stitt, Mark

    2015-05-01

    Protein synthesis and degradation represent substantial costs during plant growth. To obtain a quantitative measure of the rate of protein synthesis and degradation, we supplied (13)CO2 to intact Arabidopsis (Arabidopsis thaliana) Columbia-0 plants and analyzed enrichment in free amino acids and in amino acid residues in protein during a 24-h pulse and 4-d chase. While many free amino acids labeled slowly and incompletely, alanine showed a rapid rise in enrichment in the pulse and a decrease in the chase. Enrichment in free alanine was used to correct enrichment in alanine residues in protein and calculate the rate of protein synthesis. The latter was compared with the relative growth rate to estimate the rate of protein degradation. The relative growth rate was estimated from sequential determination of fresh weight, sequential images of rosette area, and labeling of glucose in the cell wall. In an 8-h photoperiod, protein synthesis and cell wall synthesis were 3-fold faster in the day than at night, protein degradation was slow (3%-4% d(-1)), and flux to growth and degradation resulted in a protein half-life of 3.5 d. In the starchless phosphoglucomutase mutant at night, protein synthesis was further decreased and protein degradation increased, while cell wall synthesis was totally inhibited, quantitatively accounting for the inhibition of growth in this mutant. We also investigated the rates of protein synthesis and degradation during leaf development, during growth at high temperature, and compared synthesis rates of Rubisco large and small subunits of in the light and dark. © 2015 American Society of Plant Biologists. All Rights Reserved.

  12. A metagenomics approach to evaluate the impact of dietary supplementation with Ascophyllum nodosum or Laminaria digitata on rumen function in rusitec fermenters

    Directory of Open Access Journals (Sweden)

    Alejandro eBelanche

    2016-03-01

    Full Text Available There is an increasing need to identify alternative feeds for livestock that do not compete with foods for humans. Seaweed might provide such a resource, but there is limited information available on its value as an animal feed. Here we use a multi-omics approach to investigate the value of two brown seaweeds, Ascophyllum nodosum (ASC and Laminaria digitata (LAM, as alternative feeds for ruminants. These seaweeds were supplemented at 5% inclusion rate into a control diet (CON in a rumen simulation fermenter. The seaweeds had no substantial effect on rumen fermentation, feed degradability or methane emissions. Concentrations of total bacteria, anaerobic fungi, biodiversity indices and abundances of the main bacterial and methanogen genera were also unaffected. However, species-specific effects of brown seaweed on the rumen function were noted: ASC promoted a substantial decrease in N degradability (-24% due to its high phlorotannins content. Canonical correspondence analysis of the bacterial community revealed that low N availability led to a change in the structure of the bacterial community. ASC also decreased the concentration of Escherichia coli O157:H7 post-inoculation. In contrast, LAM which has a much lower phlorotannin content did not cause detrimental effects on N degradability nor modified the structure of the bacterial community in comparison to CON. This adaptation of the microbial community to LAM diets led to a greater microbial ability to digest xylan (+70% and carboxy-methyl-cellulose (+41%. These differences among brown seaweeds resulted in greater microbial protein synthesis (+15% and non-ammonia N flow (+11% in LAM than in ASC diets and thus should led to a greater amino acid supply to the intestine of the animal. In conclusion, it was demonstrated that incorporation of brown seaweed into the diet can be considered as a suitable nutritional strategy for ruminants; however special care must be taken with those seaweeds with high

  13. Delaying aging and the aging-associated decline in protein homeostasis by inhibition of tryptophan degradation

    Science.gov (United States)

    van der Goot, Annemieke T.; Zhu, Wentao; Vázquez-Manrique, Rafael P.; Seinstra, Renée I.; Dettmer, Katja; Michels, Helen; Farina, Francesca; Krijnen, Jasper; Melki, Ronald; Buijsman, Rogier C.; Ruiz Silva, Mariana; Thijssen, Karen L.; Kema, Ido P.; Neri, Christian; Oefner, Peter J.; Nollen, Ellen A. A.

    2012-01-01

    Toxicity of aggregation-prone proteins is thought to play an important role in aging and age-related neurological diseases like Parkinson and Alzheimer’s diseases. Here, we identify tryptophan 2,3-dioxygenase (tdo-2), the first enzyme in the kynurenine pathway of tryptophan degradation, as a metabolic regulator of age-related α-synuclein toxicity in a Caenorhabditis elegans model. Depletion of tdo-2 also suppresses toxicity of other heterologous aggregation-prone proteins, including amyloid-β and polyglutamine proteins, and endogenous metastable proteins that are sensors of normal protein homeostasis. This finding suggests that tdo-2 functions as a general regulator of protein homeostasis. Analysis of metabolite levels in C. elegans strains with mutations in enzymes that act downstream of tdo-2 indicates that this suppression of toxicity is independent of downstream metabolites in the kynurenine pathway. Depletion of tdo-2 increases tryptophan levels, and feeding worms with extra l-tryptophan also suppresses toxicity, suggesting that tdo-2 regulates proteotoxicity through tryptophan. Depletion of tdo-2 extends lifespan in these worms. Together, these results implicate tdo-2 as a metabolic switch of age-related protein homeostasis and lifespan. With TDO and Indoleamine 2,3-dioxygenase as evolutionarily conserved human orthologs of TDO-2, intervening with tryptophan metabolism may offer avenues to reducing proteotoxicity in aging and age-related diseases. PMID:22927396

  14. Manipulation of rumen ecology by dietary lemongrass (Cymbopogon citratus Stapf.) powder supplementation.

    Science.gov (United States)

    Wanapat, M; Cherdthong, A; Pakdee, P; Wanapat, S

    2008-12-01

    This experiment was conducted to investigate the effect of lemongrass [Cymbopogon citratus (DC.) Stapf.] powder (LGP) on rumen ecology, rumen microorganisms, and digestibility of nutrients. Four ruminally fistulated crossbred (Brahman native) beef cattle were randomly assigned according to a 4 x 4 Latin square design. The dietary treatments were LGP supplementation at 0, 100, 200, and 300 g/d with urea-treated rice straw (5%) fed to allow ad libitum intake. Digestibilities of DM, ether extract, and NDF were significantly different among treatments and were greatest at 100 g/d of supplementation. However, digestibility of CP was decreased with LGP supplementation (P 0.05). Total viable bacteria, amylolytic bacteria, and cellulolytic bacteria were significantly different among treatments and were greatest at 100 g/d of supplementation (4.7 x 10(9), 1.7 x 10(7), and 2.0 x 10(9) cfu/mL, respectively). Protozoal populations were significantly decreased by LGP supplementation. In addition, efficiency of rumen microbial N synthesis based on OM truly digested in the rumen was enriched by LGP supplementation, especially at 100 g/d (34.2 g of N/kg of OM truly digested in the rumen). Based on this study, it could be concluded that supplementation of LGP at 100 g/d improved digestibilities of nutrients, rumen microbial population, and microbial protein synthesis efficiency, thus improving rumen ecology in beef cattle.

  15. HECTD3 Mediates an HSP90-Dependent Degradation Pathway for Protein Kinase Clients

    Directory of Open Access Journals (Sweden)

    Zhaobo Li

    2017-06-01

    Full Text Available Inhibition of the ATPase cycle of the HSP90 chaperone promotes ubiquitylation and proteasomal degradation of its client proteins, which include many oncogenic protein kinases. This provides the rationale for HSP90 inhibitors as cancer therapeutics. However, the mechanism by which HSP90 ATPase inhibition triggers ubiquitylation is not understood, and the E3 ubiquitin ligases involved are largely unknown. Using a siRNA screen, we have identified components of two independent degradation pathways for the HSP90 client kinase CRAF. The first requires CUL5, Elongin B, and Elongin C, while the second requires the E3 ligase HECTD3, which is also involved in the degradation of MASTL and LKB1. HECTD3 associates with HSP90 and CRAF in cells via its N-terminal DOC domain, which is mutationally disrupted in tumor cells with activated MAP kinase signaling. Our data implicate HECTD3 as a tumor suppressor modulating the activity of this important oncogenic signaling pathway.

  16. Particulate matter air pollution disrupts endothelial cell barrier via calpain-mediated tight junction protein degradation

    Directory of Open Access Journals (Sweden)

    Wang Ting

    2012-08-01

    Full Text Available Abstract Background Exposure to particulate matter (PM is a significant risk factor for increased cardiopulmonary morbidity and mortality. The mechanism of PM-mediated pathophysiology remains unknown. However, PM is proinflammatory to the endothelium and increases vascular permeability in vitro and in vivo via ROS generation. Objectives We explored the role of tight junction proteins as targets for PM-induced loss of lung endothelial cell (EC barrier integrity and enhanced cardiopulmonary dysfunction. Methods Changes in human lung EC monolayer permeability were assessed by Transendothelial Electrical Resistance (TER in response to PM challenge (collected from Ft. McHenry Tunnel, Baltimore, MD, particle size >0.1 μm. Biochemical assessment of ROS generation and Ca2+ mobilization were also measured. Results PM exposure induced tight junction protein Zona occludens-1 (ZO-1 relocation from the cell periphery, which was accompanied by significant reductions in ZO-1 protein levels but not in adherens junction proteins (VE-cadherin and β-catenin. N-acetyl-cysteine (NAC, 5 mM reduced PM-induced ROS generation in ECs, which further prevented TER decreases and atteneuated ZO-1 degradation. PM also mediated intracellular calcium mobilization via the transient receptor potential cation channel M2 (TRPM2, in a ROS-dependent manner with subsequent activation of the Ca2+-dependent protease calpain. PM-activated calpain is responsible for ZO-1 degradation and EC barrier disruption. Overexpression of ZO-1 attenuated PM-induced endothelial barrier disruption and vascular hyperpermeability in vivo and in vitro. Conclusions These results demonstrate that PM induces marked increases in vascular permeability via ROS-mediated calcium leakage via activated TRPM2, and via ZO-1 degradation by activated calpain. These findings support a novel mechanism for PM-induced lung damage and adverse cardiovascular outcomes.

  17. In Vitro Ruminal Degradability of Soybean Meal Protein Protected with Natural Tannin

    Science.gov (United States)

    Prasetiyono, B. W. H. E.; Subrata, A.; Tampoebolon, B. I. M.; Surono; Widiyanto

    2018-02-01

    The influence of tannin from tea waste and gambier as natural tannin sources on ruminal protein degradability was studied in this investigation. The soybean meal was used as protein source in this investigation. There were three treatments in this investigation mainly without protection (NT); protection with tea waste (Tt); and protection with gambier (Tg). The measured parameters consisted of in vitro dry matter digestibility (IVDMD), in vitro organic matter digestibility (IVOMD), and the ruminal fermentation characteristics. Results of this experiment showed that protection with tannin from tea waste as well as gambier increased (pRUP) in Tt and Tg group was higher than that in NT group (66.29 and 69.20 vs 51.10%). The ruminal protozoa population decreased (pRUP. The ruminal protozoa population and ammonia concentration, on the other hand, were decreased by tannin protection from those tannin natural sources. The natural tannin from gambier was the most effective protection agent for soybean meal protein.

  18. Differential Regulation of Telomerase Reverse Transcriptase Promoter Activation and Protein Degradation by Histone Deacetylase Inhibition.

    Science.gov (United States)

    Qing, Hua; Aono, Jun; Findeisen, Hannes M; Jones, Karrie L; Heywood, Elizabeth B; Bruemmer, Dennis

    2016-06-01

    Telomerase reverse transcriptase (TERT) maintains telomeres and is rate limiting for replicative life span. While most somatic tissues silence TERT transcription resulting in telomere shortening, cells derived from cancer or cardiovascular diseases express TERT and activate telomerase. In the present study, we demonstrate that histone deacetylase (HDAC) inhibition induces TERT transcription and promoter activation. At the protein level in contrast, HDAC inhibition decreases TERT protein abundance through enhanced degradation, which decreases telomerase activity and induces senescence. Finally, we demonstrate that HDAC inhibition decreases TERT expression during vascular remodeling in vivo. These data illustrate a differential regulation of TERT transcription and protein stability by HDAC inhibition and suggest that TERT may constitute an important target for the anti-proliferative efficacy of HDAC inhibitors. © 2015 Wiley Periodicals, Inc.

  19. Enhanced biogas yield from energy crops with rumen anaerobic fungi

    Energy Technology Data Exchange (ETDEWEB)

    Prochazka, Jindrich; Zabranska, Jana; Dohanyos, Michal [Department of Water Technology and Environmental Engineering, Faculty of Environmental Technology, Institute of Chemical Technology in Prague, Prague (Czech Republic); Mrazek, Jakub; Strosova, Lenka; Fliegerova, Katerina [Laboratory of Anaerobic Microbiology, Institute of Animal Physiology and Genetics, CAS, v.v.i., Prague (Czech Republic)

    2012-06-15

    Anaerobic fungi (AF) are able to degrade crop substrates with higher efficiency than commonly used anaerobic bacteria. The aim of this study was to investigate ways of use of rumen AF to improve biogas production from energy crops under laboratory conditions. In this study, strains of AF isolated from feces or rumen fluid of cows and deer were tested for their ability to integrate into the anaerobic bacterial ecosystem used for biogas production, in order to improve degradation of substrate polysaccharides and consequently the biogas yield. Batch culture, fed batch culture, and semicontinuous experiments have been performed using anaerobic sludge from pig slurry fermentation and different kinds of substrates (celluloses, maize, and grass silage) inoculated by different genera of AF. All experiments showed a positive effect of AF on the biogas yield and quality. AF improved the biogas production by 4-22%, depending on the substrate and AF species used. However, all the cultivation experiments indicated that rumen fungi do not show long-term survival in fermenters with digestate from pig slurry. The best results were achieved during fed batch experiment with fungal culture Anaeromyces (KF8), in which biogas production was enhanced during the whole experimental period of 140 days. This result has not been achieved in semicontinuous experiment, where increment in biogas production in fungal enriched reactor was only 4% after 42 days. (copyright 2012 Wiley-VCH Verlag GmbH and Co. KGaA, Weinheim)

  20. A critical role for protein degradation in the nucleus accumbens core in cocaine reward memory.

    Science.gov (United States)

    Ren, Zhen-Yu; Liu, Meng-Meng; Xue, Yan-Xue; Ding, Zeng-Bo; Xue, Li-Fen; Zhai, Suo-Di; Lu, Lin

    2013-04-01

    The intense associative memories that develop between cocaine-paired contexts and rewarding stimuli contribute to cocaine seeking and relapse. Previous studies have shown impairment in cocaine reward memories by manipulating a labile state induced by memory retrieval, but the mechanisms that underlie the destabilization of cocaine reward memory are unknown. In this study, using a Pavlovian cocaine-induced conditioned place preference (CPP) procedure in rats, we tested the contribution of ubiquitin-proteasome system-dependent protein degradation in destabilization of cocaine reward memory. First, we found that polyubiquitinated protein expression levels and polyubiquitinated N-ethylmaleimide-sensitive fusion (NSF) markedly increased 15 min after retrieval while NSF protein levels decreased 1 h after retrieval in the synaptosomal membrane fraction in the nucleus accumbens (NAc) core. We then found that infusion of the proteasome inhibitor lactacystin into the NAc core prevented the impairment of memory reconsolidation induced by the protein synthesis inhibitor anisomycin and reversed the effects of anisomycin on NSF and glutamate receptor 2 (GluR2) protein levels in the synaptosomal membrane fraction in the NAc core. We also found that lactacystin infusion into the NAc core but not into the shell immediately after extinction training sessions inhibited CPP extinction and reversed the extinction training-induced decrease in NSF and GluR2 in the synaptosomal membrane fraction in the NAc core. Finally, infusions of lactacystin by itself into the NAc core immediately after each training session or before the CPP retrieval test had no effect on the consolidation and retrieval of cocaine reward memory. These findings suggest that ubiquitin-proteasome system-dependent protein degradation is critical for retrieval-induced memory destabilization.

  1. L-Alanylglutamine inhibits signaling proteins that activate protein degradation, but does not affect proteins that activate protein synthesis after an acute resistance exercise.

    Science.gov (United States)

    Wang, Wanyi; Choi, Ran Hee; Solares, Geoffrey J; Tseng, Hung-Min; Ding, Zhenping; Kim, Kyoungrae; Ivy, John L

    2015-07-01

    Sustamine™ (SUS) is a dipeptide composed of alanine and glutamine (AlaGln). Glutamine has been suggested to increase muscle protein accretion; however, the underlying molecular mechanisms of glutamine on muscle protein metabolism following resistance exercise have not been fully addressed. In the present study, 2-month-old rats climbed a ladder 10 times with a weight equal to 75 % of their body mass attached at the tail. Rats were then orally administered one of four solutions: placebo (PLA-glycine = 0.52 g/kg), whey protein (WP = 0.4 g/kg), low dose of SUS (LSUS = 0.1 g/kg), or high dose of SUS (HSUS = 0.5 g/kg). An additional group of sedentary (SED) rats was intubated with glycine (0.52 g/kg) at the same time as the ladder-climbing rats. Blood samples were collected immediately after exercise and at either 20 or 40 min after recovery. The flexor hallucis longus (FHL), a muscle used for climbing, was excised at 20 or 40 min post exercise and analyzed for proteins regulating protein synthesis and degradation. All supplements elevated the phosphorylation of FOXO3A above SED at 20 min post exercise, but only the SUS supplements significantly reduced the phosphorylation of AMPK and NF-kB p65. SUS supplements had no effect on mTOR signaling, but WP supplementation yielded a greater phosphorylation of mTOR, p70S6k, and rpS6 compared with PLA at 20 min post exercise. However, by 40 min post exercise, phosphorylation of mTOR and rpS6 in PLA had risen to levels not different than WP. These results suggest that SUS blocks the activation of intracellular signals for MPB, whereas WP accelerates mRNA translation.

  2. In vitro evaluation of different varieties of maize fodder for their methane generation potential and digestibility with goat rumen liquor.

    Science.gov (United States)

    Vaswani, Shalini; Kumar, Ravindra; Kumar, Vinod; Roy, Debashis; Kumar, Muneendra

    2016-11-01

    To evaluate the methane generation potential and digestibility of different (normal and three high-quality protein maize [HQPM]) varieties of maize fodder with goat rumen liquor in vitro . Methane production potential and digestibility of different varieties of maize fodder were tested in in vitro gas production test. Seven varieties of maize, four normal (HTHM 5101, DHM 117, HM 5, and Shaktiman/900 M Gold), and three high-quality protein (HQPM 5, HQPM 7, and HQPM 9/Vivek) were grown in different plots under the same environmental and agro-climatic conditions. Fodders were harvested at 45-50 days of sowing, and the representative samples of fodder from different varieties of maize were collected for analysis. Dried and grinded form of these maize fodder varieties was tested for gas, methane, and digestibility using goat rumen microflora in in vitro gas syringes. Gas production (ml/g dry matter [DM]) was highest for HM5 variety (97.66, whereas lowest for HQPM 9 variety (64.22). Gas production (ml/g degraded DM [DDM]) and methane (%) were statistically similar in different varieties of maize fodder. The methane production expressed as ml/g DM and ml/g DDM was significantly (pproduction.

  3. CNPY2 inhibits MYLIP-mediated AR protein degradation in prostate cancer cells.

    Science.gov (United States)

    Ito, Saya; Ueno, Akihisa; Ueda, Takashi; Nakagawa, Hideo; Taniguchi, Hidefumi; Kayukawa, Naruhiro; Fujihara-Iwata, Atsuko; Hongo, Fumiya; Okihara, Koji; Ukimura, Osamu

    2018-04-03

    The androgen receptor (AR) is a ligand-dependent transcription factor that promotes prostate cancer (PC) cell growth through control of target gene expression. This report suggests that Canopy FGF signaling regulator 2 (CNPY2) controls AR protein levels in PC cells. We found that AR was ubiquitinated by an E3 ubiquitin ligase, myosin regulatory light chain interacting protein (MYLIP) and then degraded through the ubiquitin-proteasome pathway. CNPY2 decreased the ubiquitination activity of MYLIP by inhibition of interaction between MYLIP and UBE2D1, an E2 ubiquitin ligase. CNPY2 up-regulated gene expression of AR target genes such as KLK3 gene which encodes the prostate specific antigen (PSA) and promoted cell growth of PC cells. The cell growth inhibition by CNPY2 knockdown was rescued by AR overexpression. Furthermore, positive correlation of expression levels between CNPY2 and AR/AR target genes was observed in tissue samples from human prostate cancer patients. Together, these results suggested that CNPY2 promoted cell growth of PC cells by inhibition of AR protein degradation through MYLIP-mediated AR ubiquitination.

  4. Saturated fatty acid palmitate negatively regulates autophagy by promoting ATG5 protein degradation in meniscus cells.

    Science.gov (United States)

    Mallik, Aritra; Yammani, Raghunatha R

    2018-07-20

    Obesity and associated metabolic factors are major risk factors for the development of osteoarthritis. Previously, we have shown that the free fatty acid palmitate induces endoplasmic reticulum (ER) stress and induces apoptosis in meniscus cells. However, the molecular mechanisms involved in these effects are not clearly understood. In our current study, we found that palmitate inhibits autophagy by modulating the protein levels of autophagy-related genes-5 (ATG5) that is associated with decreased lipidation of LC3 and increased activation of cleaved caspase 3. Pretreatment of meniscus cells with 4-phenyl butyric acid, a small molecule chemical chaperone that alleviates ER stress, or with MG-132, a proteasome inhibitor, restored normal levels of ATG5 and autophagosome formation, and decreased expression of cleaved caspase 3. Thus, our data suggest that palmitate downregulates autophagy in meniscus cells by degrading ATG5 protein via ER-associated protein degradation, and thus promotes apoptosis. This is the first study to demonstrate that palmitate-induced endoplasmic reticulum stress negatively regulates autophagy. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

  5. NAD(P)H quinone oxidoreductase 1 inhibits the proteasomal degradation of homocysteine-induced endoplasmic reticulum protein

    Energy Technology Data Exchange (ETDEWEB)

    Maeda, Tomoji, E-mail: t-maeda@nichiyaku.ac.jp [Department of Neuroscience, School of Pharmacy, Iwate Medical University, 2-1-1 Nishitokuta, Yahaba-Cho, Shiwagun, Iwate, 028-3603 (Japan); Tanabe-Fujimura, Chiaki; Fujita, Yu; Abe, Chihiro; Nanakida, Yoshino; Zou, Kun; Liu, Junjun; Liu, Shuyu [Department of Neuroscience, School of Pharmacy, Iwate Medical University, 2-1-1 Nishitokuta, Yahaba-Cho, Shiwagun, Iwate, 028-3603 (Japan); Nakajima, Toshihiro [Institute of Medical Science, Tokyo Medical University, 6-1-1 Shinjyuku, Shinjyuku, Tokyo, Tokyo, 160-8402 (Japan); Komano, Hiroto, E-mail: hkomano@iwate-med.ac.jp [Department of Neuroscience, School of Pharmacy, Iwate Medical University, 2-1-1 Nishitokuta, Yahaba-Cho, Shiwagun, Iwate, 028-3603 (Japan)

    2016-05-13

    Homocysteine-induced endoplasmic reticulum (ER) protein (Herp) is an ER stress-inducible key regulatory component of ER-associated degradation (ERAD) that has been implicated in insulin hypersecretion in diabetic mouse models. Herp expression is tightly regulated. Additionally, Herp is a highly labile protein and interacts with various proteins, which are characteristic features of ubiquitinated protein. Previously, we reported that ubiquitination is not required for Herp degradation. In addition, we found that the lysine residues of Herp (which are ubiquitinated by E3 ubiquitin ligase) are not sufficient for regulation of Herp degradation. In this study, we found that NAD(P)H quinone oxidoreductase 1 (NQO1)-mediated targeting of Herp to the proteasome was involved in Herp degradation. In addition, we found that Herp protein levels were markedly elevated in synoviolin-null cells. The E3 ubiquitin ligase synoviolin is a central component of ERAD and is involved in the degradation of nuclear factor E2-related factor-2 (Nrf2), which regulates cellular reactive oxygen species. Additionally, NQO1 is a target of Nrf2. Thus, our findings indicated that NQO1 could stabilize Herp protein expression via indirect regulation of synoviolin. -- Highlights: •Herp interacts with NQO1. •NQO1 regulates Herp degradation.

  6. NAD(P)H quinone oxidoreductase 1 inhibits the proteasomal degradation of homocysteine-induced endoplasmic reticulum protein

    International Nuclear Information System (INIS)

    Maeda, Tomoji; Tanabe-Fujimura, Chiaki; Fujita, Yu; Abe, Chihiro; Nanakida, Yoshino; Zou, Kun; Liu, Junjun; Liu, Shuyu; Nakajima, Toshihiro; Komano, Hiroto

    2016-01-01

    Homocysteine-induced endoplasmic reticulum (ER) protein (Herp) is an ER stress-inducible key regulatory component of ER-associated degradation (ERAD) that has been implicated in insulin hypersecretion in diabetic mouse models. Herp expression is tightly regulated. Additionally, Herp is a highly labile protein and interacts with various proteins, which are characteristic features of ubiquitinated protein. Previously, we reported that ubiquitination is not required for Herp degradation. In addition, we found that the lysine residues of Herp (which are ubiquitinated by E3 ubiquitin ligase) are not sufficient for regulation of Herp degradation. In this study, we found that NAD(P)H quinone oxidoreductase 1 (NQO1)-mediated targeting of Herp to the proteasome was involved in Herp degradation. In addition, we found that Herp protein levels were markedly elevated in synoviolin-null cells. The E3 ubiquitin ligase synoviolin is a central component of ERAD and is involved in the degradation of nuclear factor E2-related factor-2 (Nrf2), which regulates cellular reactive oxygen species. Additionally, NQO1 is a target of Nrf2. Thus, our findings indicated that NQO1 could stabilize Herp protein expression via indirect regulation of synoviolin. -- Highlights: •Herp interacts with NQO1. •NQO1 regulates Herp degradation.

  7. Novel function of the endoplasmic reticulum degradation-enhancing α-mannosidase-like proteins in the human hepatitis B virus life cycle, mediated by the middle envelope protein.

    Science.gov (United States)

    Lazar, Catalin; Uta, Mihaela; Petrescu, Stefana Maria; Branza-Nichita, Norica

    2017-02-01

    Cells replicating the human hepatitis B virus (HBV) express high levels of degradation-enhancing α-mannosidase-like proteins (EDEMs), a family of proteins involved in the endoplasmic reticulum associated degradation, one of the pathways activated during the unfolded protein response. Owing to their α-1,2 mannosidase activity, the EDEM1-3 proteins are able to process the N-linked glycans of misfolded or incompletely folded proteins, providing the recognition signal for their subsequent degradation. The HBV small (S), medium (M), and large (L) surface proteins bear an N-linked glycosylation site in the common S domain that is partially occupied in all proteins. The M protein contains an additional site in its preS2 domain, which is always functional. Here, we report that these oligosaccharides are processed by EDEMs, more efficiently by EDEM3, which induces degradation of L and S proteins, accompanied by a reduction of subviral particles production. In striking contrast, M not only is spared from degradation but its trafficking is also accelerated leading to an improved secretion. This unusual behavior of the M protein requires strictly the mannose trimming of the preS2 N-linked glycan. Furthermore, we show that HBV secretion is significantly inhibited under strong endoplasmic reticulum stress conditions when M expression is prevented by mutagenesis of the viral genome. These observations unfold unique properties of the M protein in the HBV life cycle during unfolded protein response and point to alternative mechanisms employed by EDEMs to alleviate this stress in case of necessity by promoting glycoprotein trafficking rather than degradation. © 2016 John Wiley & Sons Ltd.

  8. Rumen bacteria at work: bioaugmentation strategies to enhance biogas production from cow manure.

    Science.gov (United States)

    Ozbayram, E G; Akyol, Ç; Ince, B; Karakoç, C; Ince, O

    2018-02-01

    To investigate the effects of different bioaugmentation strategies for enhancing the biogas production from cow manure and evaluate microbial community patterns. Co-inoculation with cow rumen fluid and cow rumen-derived enriched microbial consortia was evaluated in anaerobic batch tests at 36°C and 41°C. Singular addition of both rumen fluid and enriched bioaugmentation culture had a promising enhancement on methane yields; however, the highest methane yield (311 ml CH 4 per gram VS at 41°C) was achieved when the anaerobic seed sludge was co-inoculated together with rumen fluid and enriched bioaugmentation culture. Bacterial community profiles were investigated by Ion PGM Platform, and specific lignocellulolytic bacteria dynamics in batch tests were assessed by qPCR. The temperature had minor effects on the abundance of bacterial community; in which Bacteroidetes and Firmicutes were the most abundant phyla in all digesters. Furthermore, Rikenellaceae, Clostridiaceae, Porphyromonadaceae, Bacteroidaceae and Ruminococcaceae played a crucial role during the anaerobic degradation of cow manure. There was an important impact of Firmicutes flavefaciens and Ruminococcus albus at 41°C, which in turn positively affected the methane production. The degree of enhancement in biogas production can be upgraded by the co-inoculation of rumen-derived bioaugmentation culture with anaerobic seed sludge with high methanogenic activity. A close look at the biotic interactions and their associations with abiotic factors might be valuable for evaluating rumen-related bioaugmentation applications. © 2017 The Society for Applied Microbiology.

  9. High genetic diversity and different distributions of glycosyl hydrolase family 10 and 11 xylanases in the goat rumen.

    Directory of Open Access Journals (Sweden)

    Guozeng Wang

    Full Text Available BACKGROUND: The rumen harbors a complex microbial ecosystem for efficient hydrolysis of plant polysaccharides which are the main constituent of the diet. Xylanase is crucial for hemicellulose hydrolysis and plays an important role in the plant cell wall degradation. Xylanases of ruminal strains were widely studied, but few studies have focused on their diversity in rumen microenvironment. METHODOLOGY/PRINCIPAL FINDINGS: We explored the genetic diversity of xylanases belonging to two major glycosyl hydrolase families (GH 10 and 11 in goat rumen contents by analyzing the amplicons generated with two degenerate primer sets. Fifty-two distinct GH 10 and 35 GH 11 xylanase gene fragments (similarity <95% were retrieved, and most had low identities with known sequences. Based on phylogenetic analysis, all GH 10 xylanase sequences fell into seven clusters, and 88.5% of them were related to xylanases from Bacteroidetes. Five clusters of GH 11 xylanase sequences were identified. Of these, 85.7% were related to xylanases from Firmicutes, and 14.3% were related to those of rumen fungi. Two full-length xylanase genes (one for each family were directly cloned and expressed in Escherichia coli. Both the recombinant enzymes showed substantial xylanase activity, and were purified and characterized. Combined with the results of sheep rumen, Bacteroidetes and Firmicutes are the two major phyla of xylan-degrading microorganisms in rumen, which is distinct from the representatives of other environments such as soil and termite hindgut, suggesting that xylan-degrading microorganisms are environment specific. CONCLUSION/SIGNIFICANCE: The numerous new xylanase genes suggested the functional diversity of xylanase in the rumen microenvironment which may have great potential applications in industry and agriculture. The phylogenetic diversity and different distributions of xylanase genes will help us understand their roles in plant cell wall degradation in the rumen

  10. Degradation characteristics of urea and lime treated groundnut ...

    African Journals Online (AJOL)

    This research was conducted to investigate the chemical composition and rumen degradation characteristics of treated groundnut shells (GNS) based diets in the rumen. It was carried out in the Teaching and Research farm of the Department of Animal Science A.B.U. Zaria. Three fistulated Yankasa rams with average ...

  11. GCK-MODY diabetes associated with protein misfolding, cellular self-association and degradation.

    Science.gov (United States)

    Negahdar, Maria; Aukrust, Ingvild; Johansson, Bente B; Molnes, Janne; Molven, Anders; Matschinsky, Franz M; Søvik, Oddmund; Kulkarni, Rohit N; Flatmark, Torgeir; Njølstad, Pål Rasmus; Bjørkhaug, Lise

    2012-11-01

    GCK-MODY, dominantly inherited mild fasting hyperglycemia, has been associated with >600 different mutations in the glucokinase (GK)-encoding gene (GCK). When expressed as recombinant pancreatic proteins, some mutations result in enzymes with normal/near-normal catalytic properties. The molecular mechanism(s) of GCK-MODY due to these mutations has remained elusive. Here, we aimed to explore the molecular mechanisms for two such catalytically 'normal' GCK mutations (S263P and G264S) in the F260-L270 loop of GK. When stably overexpressed in HEK293 cells and MIN6 β-cells, the S263P- and G264S-encoded mutations generated misfolded proteins with an increased rate of degradation (S263P>G264S) by the protein quality control machinery, and a propensity to self-associate (G264S>S263P) and form dimers (SDS resistant) and aggregates (partly Triton X-100 insoluble), as determined by pulse-chase experiments and subcellular fractionation. Thus, the GCK-MODY mutations S263P and G264S lead to protein misfolding causing destabilization, cellular dimerization/aggregation and enhanced rate of degradation. In silico predicted conformational changes of the F260-L270 loop structure are considered to mediate the dimerization of both mutant proteins by a domain swapping mechanism. Thus, similar properties may represent the molecular mechanisms for additional unexplained GCK-MODY mutations, and may also contribute to the disease mechanism in other previously characterized GCK-MODY inactivating mutations. Copyright © 2012 Elsevier B.V. All rights reserved.

  12. In vitro biodegradation of cyanotoxins in the rumen fluid of cattle.

    Science.gov (United States)

    Manubolu, Manjunath; Madawala, Samanthi R P; Dutta, Paresh C; Malmlöf, Kjell

    2014-05-08

    In countries around the Baltic Sea grazing ruminants have access to and drink, surface water from lakes, rivers and in several coastal regions. The water quality of these naturally occurring reservoirs affects performance and health of livestock. In the Baltic Sea both microcystin (MC) and nodularin (NOD) occurs as cyclic peptides and have hepatotoxic effects. Although cattle obviously have died after consuming contaminated water very little information is available as to how susceptible ruminants are to the toxins produced by cyanobacteria. The critical question as to whether the rumen microflora might constitute a protective shield is unresolved. For this reason our aim is to investigate a possible degradation rate of these toxins in rumen. The ability of rumen microorganisms to degrade certain important cyanotoxins (MC-LR, YR, RR and NOD) was studied in vitro by incubating with rumen fluid at three different concentrations (0.05, 0.5 and 5 μg/mL) for 3 h. The degradation efficiencies were determined by LC-MS (ESI) positive mode. Degradation was observed in the following order MC-RR 36%, NOD 35%, MC-RR 25% and MC-LR 8.9% at lower concentrations within 3 h. However, average degradation was observed at concentration of 0.5 μg/mL. No degradation was observed in higher concentrations for entire 3 h. The present results reveal that the degradation was both dose and time dependent. In conclusion the present results suggest that the rumen microbial flora may protect ruminants from being intoxicated by Cyanotoxins.

  13. Ubiquitin Ligase gp78 Targets Unglycosylated Prion Protein PrP for Ubiquitylation and Degradation

    OpenAIRE

    Shao, Jia; Choe, Vitnary; Cheng, Haili; Tsai, Yien Che; Weissman, Allan M.; Luo, Shiwen; Rao, Hai

    2014-01-01

    Prion protein PrP is a central player in several devastating neurodegenerative disorders, including mad cow disease and Creutzfeltd-Jacob disease. Conformational alteration of PrP into an aggregation-prone infectious form PrPSc can trigger pathogenic events. How levels of PrP are regulated is poorly understood. Human PrP is known to be degraded by the proteasome, but the specific proteolytic pathway responsible for PrP destruction remains elusive. Here, we demonstrate that the ubiquitin ligas...

  14. Detoksifikasi Mikotoksin Melalui Optimalisasi Fungsi Rumen dengan Pemberian Ragi (MYCOTOXIN DETOXIFICATION THROUGH OPTIMIZATION THE RUMEN FUNCTION BY YEAST

    Directory of Open Access Journals (Sweden)

    Dadik Pantaya

    2016-03-01

    Full Text Available Mycotoxins are toxic metabolites produced by some fungal species commonly found in food and feed,particularly in cereals. In intensive production systems, dairy cattle are commonly fed with cereal-richdiets and, consequently, are more exposed to micotoxins. Besides, such diet is often associated with ahigher risk of rumen acidosis which can also affect the performance and the helath of animal. In addition,the efficacy of microbial detoxification can be reduced during acidosis. For instance, some authors observeda decrease in the number of protozoa that are responsible for the degradation of some mycotoxins. Anotherconsequence of acidosis is the potential modification of ruminal absorption of mycotoxins, which until nowhas received scarce attention. Yeast Saccharomyces cerevisiae, probiotic additives have been shown toreduce the post-feeding drop in rumen pH and to increase the number of ruminal protozoa. This effect canbe positive in reducing the absorption and toxicity of mycotoxins in ruminantia.

  15. Two waves of proteasome-dependent protein degradation in the hippocampus are required for recognition memory consolidation.

    Science.gov (United States)

    Figueiredo, Luciana S; Dornelles, Arethuza S; Petry, Fernanda S; Falavigna, Lucio; Dargél, Vinicius A; Köbe, Luiza M; Aguzzoli, Cristiano; Roesler, Rafael; Schröder, Nadja

    2015-04-01

    Healthy neuronal function and synaptic modification require a concert of synthesis and degradation of proteins. Increasing evidence indicates that protein turnover mediated by proteasome activity is involved in long-term synaptic plasticity and memory. However, its role in different phases of memory remains debated, and previous studies have not examined the possible requirement of protein degradation in recognition memory. Here, we show that the proteasome inhibitor, lactacystin (LAC), infused into the CA1 area of the hippocampus at two specific time points during consolidation, impairs 24-retention of memory for object recognition in rats. Administration of LAC after retrieval did not affect retention. These findings provide the first evidence for a requirement of proteasome activity in recognition memory, indicate that protein degradation in the hippocampus is necessary during selective time windows of memory consolidation, and further our understanding of the role of protein turnover in memory formation. Copyright © 2015 Elsevier Inc. All rights reserved.

  16. Clofibrate-induced increases in peroxisomal proteins: effect on synthesis, degradation, and mRNA activity

    International Nuclear Information System (INIS)

    Mortensen, R.M.

    1983-01-01

    The effect of clofibrate on the polypeptide composition of peroxisomes was determined. A simple method was developed for the isolation of peroxisomes with a purity of 90-95% using sedimentation in a metrizamide gradient. The specific activities of HD did not change with clofibrate treatment so that the increases in enzyme activities are solely due to increases in protein amounts. The hepatic concentration of HD increased 63 times. The HD synthesis rate, as measured by the incorporation of [ 3 H]leucine, increased 74 times, so that the increase in the synthesis was sufficient to account for the increase in protein. Clofibrate caused no discernible change in the degradation rate of HD labeled with [ 14 C]bicarbonate. The half-life of HD was approximately 2 days. The translatable mRBA coding for HD increased 55 times. This value is not significantly different from the increase in HD protein or in HD synthesis. This observation was also true for several other peroxisomal proteins. Therefore, clofibrate causes an increase in the mRNA activity, which increases the synthesis of HD leading to an accumulation of protein and enzyme activity. The kinetics of the clofibrate-induced changes in HD synthesis rate, protein level, and enzymatic activity was analyzed using a simple model which included the half-lives of the drug, mRNA, and protein. The best fit of the model to the data gave an mRNA half-life of 10 hours and a protein half-life of 1.8 days, with no significant change by clofibrate

  17. Effects of yeast culture supplement on digestion of nutrients and rumen fermentation in cattle fed on grass silage barley diet

    Directory of Open Access Journals (Sweden)

    Pekka Huhtanen

    1991-12-01

    Full Text Available The effects of including yeast culture (Saccharomyces cerevisiae plus growth medium; 5 x 106 organisms/g on the digestion of dietary constituents in the rumen and total digestive tract were studied in a pair of monozygote twin bulls. The animals were fitted with cannulae in the rumen and in the proximal duodenum. A diet of grass silage, barley and rapeseed meal (445, 445 and 90 g/kg total dry matter (DM was fed, with and without addition of 10 g per day of yeast culture (YC, in two treatment sequences. The addition of YC had no effect on the mean values of rumen pH, ammonia N concentration or molar proportions of volatile fatty acids. Also, the postprandial changes in rumen fermentation pattern were similar when the diet did and did not contain the YC supplement. The peak concentration of lactic acid 1 h after feeding tended to be higher in cattle receiving the YC diet (13.9 v 6.0 mmol/l. Apparent digestibility of organic matter (OM (mean 0.780 and the proportion of OM digestion occurring in the rumen (mean 0.603 were not affected by YC. Likewise, there was no effect on rumen or total digestion of cell wall carbohydrates, and the results for the degradation of hay DM in the rumen and for particle-associated carboxymethylcellulase and xylanase activities indicated that YC had no effect on the rumen environment that could affect fibre digestion. Supplemental yeast did not affect the rate of microbial N synthesis (28.0 and 28.6 g/kg OM apparently digested in the rumen. The results indicate that the addition of YC to the diet is not likely to improve the efficiency of digestion and fermentation in the rumen of cattle given a diet based on grass silage and barley.

  18. Insights into the bovine rumen plasmidome

    Science.gov (United States)

    Kav, Aya Brown; Sasson, Goor; Jami, Elie; Doron-Faigenboim, Adi; Benhar, Itai; Mizrahi, Itzhak

    2012-01-01

    Plasmids are self-replicating genetic elements capable of mobilization between different hosts. Plasmids often serve as mediators of lateral gene transfer, a process considered to be a strong and sculpting evolutionary force in microbial environments. Our aim was to characterize the overall plasmid population in the environment of the bovine rumen, which houses a complex and dense microbiota that holds enormous significance for humans. We developed a procedure for the isolation of total rumen plasmid DNA, termed rumen plasmidome, and subjected it to deep sequencing using the Illumina paired-end protocol and analysis using public and custom-made bioinformatics tools. A large number of plasmidome contigs aligned with plasmids of rumen bacteria isolated from different locations and at various time points, suggesting that not only the bacterial taxa, but also their plasmids, are defined by the ecological niche. The bacterial phylum distribution of the plasmidome was different from that of the rumen bacterial taxa. Nevertheless, both shared a dominance of the phyla Firmicutes, Bacteroidetes, and Proteobacteria. Evidently, the rumen plasmidome is of a highly mosaic nature that can cross phyla. Interestingly, when we compared the functional profile of the rumen plasmidome to two plasmid databases and two recently published rumen metagenomes, it became apparent that the rumen plasmidome codes for functions, which are enriched in the rumen ecological niche and could confer advantages to their hosts, suggesting that the functional profiles of mobile genetic elements are associated with their environment, as has been previously implied for viruses. PMID:22431592

  19. FibroChip, a Functional DNA Microarray to Monitor Cellulolytic and Hemicellulolytic Activities of Rumen Microbiota

    Directory of Open Access Journals (Sweden)

    Sophie Comtet-Marre

    2018-02-01

    Full Text Available Ruminants fulfill their energy needs for growth primarily through microbial breakdown of plant biomass in the rumen. Several biotic and abiotic factors influence the efficiency of fiber degradation, which can ultimately impact animal productivity and health. To provide more insight into mechanisms involved in the modulation of fibrolytic activity, a functional DNA microarray targeting genes encoding key enzymes involved in cellulose and hemicellulose degradation by rumen microbiota was designed. Eight carbohydrate-active enzyme (CAZyme families (GH5, GH9, GH10, GH11, GH43, GH48, CE1, and CE6 were selected which represented 392 genes from bacteria, protozoa, and fungi. The DNA microarray, designated as FibroChip, was validated using targets of increasing complexity and demonstrated sensitivity and specificity. In addition, FibroChip was evaluated for its explorative and semi-quantitative potential. Differential expression of CAZyme genes was evidenced in the rumen bacterium Fibrobacter succinogenes S85 grown on wheat straw or cellobiose. FibroChip was used to identify the expressed CAZyme genes from the targeted families in the rumen of a cow fed a mixed diet based on grass silage. Among expressed genes, those encoding GH43, GH5, and GH10 families were the most represented. Most of the F. succinogenes genes detected by the FibroChip were also detected following RNA-seq analysis of RNA transcripts obtained from the rumen fluid sample. Use of the FibroChip also indicated that transcripts of fiber degrading enzymes derived from eukaryotes (protozoa and anaerobic fungi represented a significant proportion of the total microbial mRNA pool. FibroChip represents a reliable and high-throughput tool that enables researchers to monitor active members of fiber degradation in the rumen.

  20. FibroChip, a Functional DNA Microarray to Monitor Cellulolytic and Hemicellulolytic Activities of Rumen Microbiota.

    Science.gov (United States)

    Comtet-Marre, Sophie; Chaucheyras-Durand, Frédérique; Bouzid, Ourdia; Mosoni, Pascale; Bayat, Ali R; Peyret, Pierre; Forano, Evelyne

    2018-01-01

    Ruminants fulfill their energy needs for growth primarily through microbial breakdown of plant biomass in the rumen. Several biotic and abiotic factors influence the efficiency of fiber degradation, which can ultimately impact animal productivity and health. To provide more insight into mechanisms involved in the modulation of fibrolytic activity, a functional DNA microarray targeting genes encoding key enzymes involved in cellulose and hemicellulose degradation by rumen microbiota was designed. Eight carbohydrate-active enzyme (CAZyme) families (GH5, GH9, GH10, GH11, GH43, GH48, CE1, and CE6) were selected which represented 392 genes from bacteria, protozoa, and fungi. The DNA microarray, designated as FibroChip, was validated using targets of increasing complexity and demonstrated sensitivity and specificity. In addition, FibroChip was evaluated for its explorative and semi-quantitative potential. Differential expression of CAZyme genes was evidenced in the rumen bacterium Fibrobacter succinogenes S85 grown on wheat straw or cellobiose. FibroChip was used to identify the expressed CAZyme genes from the targeted families in the rumen of a cow fed a mixed diet based on grass sila