WorldWideScience

Sample records for ruminally degraded protein

  1. Short communication In situ ruminal degradability of protein feeds ...

    African Journals Online (AJOL)

    2016-12-28

    Dec 28, 2016 ... to evaluate the ruminal degradability of protein feeds with different physical forms (meal versus grain) through a meta-analysis study. A database, composed of 45 treatments from 12 studies carried out in Brazil, was evaluated to compare the degradability of soybean, sunflower, canola, and flaxseed grains ...

  2. 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. L.J. Erasmus,· J. Prinsloo1 and P.M. Botha. Animal and Dairy Science Research Institute, Private Bag X2, Irene 1675, Republic of South Africa. H.H. Meissner. Department of Livestock Science, ...

  3. 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

  4. IN VITRO RUMINAL PROTEIN DEGRADATION AND MICROBIAL PROTEIN FORMATION OF SEED LEGUMES

    Science.gov (United States)

    Seed legumes such as peas, lupins, and faba beans are important feeds for dairy cows in Europe and other regions. Ruminal protein degradability was quantified using the inhibitor in vitro (IIV) system for samples of 5 seed legumes: 2 peas (cv. Alembo and Helena), 1 white lupin (Lupinus albus, cv. Mu...

  5. Ruminal degradability and intestinal digestion of eight plant protein ...

    African Journals Online (AJOL)

    Unknown

    The extent of microbial degradation of protein supplements in the rumen ... proportion of the feed protein is available for microbial metabolism, which by difference ... 8g ) into nitrogen-free-polyester bags and incubated in the rumen of cows ...

  6. Ruminal degradability and intestinal digestion of eight plant protein ...

    African Journals Online (AJOL)

    Unknown

    dairy cattle respectively. Table 1 The crude protein (CP) content and degradation properties of dry matter (DM) and nitrogen. (N) of plant protein supplements incubated in the rumens of Jersey cows. DM Loss properties. CP. N Loss properties. Source a (g/kg) b (g/kg) c (/h). (g/kg DM) a (g/kg) b (g/kg) c (/h). Canola. 271. 630.

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

    African Journals Online (AJOL)

    A database, composed of 45 treatments from 12 studies carried out in Brazil, was evaluated to compare the degradability of soybean, sunflower, canola, and flaxseed grains with the meal forms of soybean, peanut, sunflower, cottonseed, and corn gluten. The degradation parameters of dry matter did not differ significantly ...

  8. Ruminal degradation of protein : implications for polluting emissions

    OpenAIRE

    Vanegas Ruiz, Jorge Leonardo

    2016-01-01

    Los rumiantes contribuyen a la emisión de gases de efecto invernadero, principalmente a través del metano (CH4) producido por la fermentación ruminal, que tiene un efecto invernadero entre 21 y 25 veces más potente que el CO2. Por esta razón, en los últimos años se han desarrollado numerosas investigaciones que se han centrado en el análisis de los factores dietéticos que afectan a la producción de este gas, si bien apenas se ha investigado el efecto de las características de las fuentes de n...

  9. Manipulating Protein Degradability in the Rumen to Support Higher Ruminant Production

    Directory of Open Access Journals (Sweden)

    Budi Haryanto

    2014-09-01

    Full Text Available Dietary protein is digested to a certain extent in the rumen causing decreases its potency as source of amino acids for the animal. Dietary protein should mostly reach the intestines where the protein digestion takes place and absorption occurs in the form of intact amino acids and subsequently becomes nutrient deposition in muscles or milk. The higher muscle or milk protein synthesis, the higher the protein in the products of animal, as long as energy for the metabolism is available. Strategies of feeding rumen degradable versus undegradable protein in ruminant have become a research interest for decades. Technologies of dietary protein protection to reduce its degradability in the rumen by heating, chelating or coating have been developed.

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

    African Journals Online (AJOL)

    The rate constant for outflow was correlated with level of DM intake (r2 = 0.87). Lag phase prior to degrada- tion of crude ... at a level of 7% for the whole maize and 11% for the maize meal and flaked maize diets. Urea was .... have been affected by both the particle type and the math- ematical model we used. Crude protein ...

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

    Directory of Open Access Journals (Sweden)

    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.

  12. Simplified in situ method for estimating ruminal dry matter and protein degradability of concentrates.

    Science.gov (United States)

    Olaisen, V; Mejdell, T; Volden, H; Nesse, N

    2003-02-01

    In this study, dry matter and crude protein in situ degradation data from different concentrate feeds were used to test the accuracy of effective degradability (ED) measures when using reduced ruminal incubation times compared with models based on seven or eight incubation times. The ED was estimated both with and without correction for nylon bag particle loss. The crude protein ED corrected for particle loss of the calibration data set was widely distributed in a range from 16 to 90% with an overall mean value of 60.4%, and the dry matter ED was distributed in the range from 22.7 to 80.7%, with a mean value of 56.9%. The simplified method was developed based on bilinear regression models where all combinations of one to three disappearance values were tested to find the optimal time point combinations to estimate ED. Bilinear regression models based on two and three ruminal incubation times gave similar estimates to a standard in situ method over a wide range of passage rates both for the data set used to parameterize the models and the independent data set used to evaluate the models. Using two incubation times, the bilinear model based on 4 and 24 h gave the most accurate estimates, and the models based on 2, 8, and 24 h for uncorrected data and 4, 8, and 24 h for corrected data were most accurate of the three time points bilinear models. The number of nylon bags used by these models was reduced by 58 to 78% compared with the standard in situ method, and the total incubation time needed was substantially reduced.

  13. Protein degradation by ruminal microorganisms from sheep fed dietary supplements of urea, casein, or albumin.

    Science.gov (United States)

    Wallace, R J; Broderick, G A; Brammall, M L

    1987-01-01

    Ruminal fluid from sheep fed hay plus concentrate diets containing 1.8% urea, 6% casein, or 6% egg albumin had proteolytic activities of 4.12, 3.02, or 4.00 mg of [14C]casein hydrolyzed ml-1 h-1, respectively. Dietary albumin had no effect on the rate of albumin breakdown relative to that of casein (0.06). Greater numbers of highly proteolytic bacteria, mainly Butyrivibrio spp., were isolated from the rumens of sheep receiving albumin. Albumin hydrolysis by these isolates was even slower relative to that of casein (0.03) than in ruminal fluid and was similar to that found in isolates from urea- and casein-fed sheep. Hence, there appears to be no mechanism by which ruminal bacteria can alter their proteolytic activity to utilize resistant soluble protein more effectively. PMID:3579280

  14. Effect of conventional and extrusion pelleting on in situ ruminal degradability of starch, protein, and fibre in cattle

    DEFF Research Database (Denmark)

    Razzaghi, Ali; Larsen, Mogens; Lund, Peter

    2016-01-01

    Rumen degradability kinetics was investigated for pelleted compound concentrates differing in physical functional properties of pellets. Six compound concentrate meals were produced containing either 100% wheat, 100% maize, 50% wheat+50% soybean meal (SBM; as is basis), 50% maize+50% SBM, 50% wheat......+50% sugar beet pulp (SBP), or 50% maize+50% SBP. Meals were pelleted by either conventional pelleting, or by cooking extrusion using two distinct settings giving pellets with either high density (HD) or low density (LD). Ruminal degradation of starch, crude protein (CP) and NDF, and intestinal......>Pelleting>Meal). In contradiction, ESD for pure wheat and wheat mixtures was reduced, though differences were minor. Conventional pelleting reduced the effective protein degradability (EPD) for pure wheat, but extrusion did not further affect the EPD. In contrast, the most intense processing with extrusion LD increased EPD...

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

    Directory of Open Access Journals (Sweden)

    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.

  16. Predicting the effect of proteolysis on ruminal crude protein degradation of legume and grass silages using near-infrared reflectance spectroscopy.

    Science.gov (United States)

    Hoffman, P C; Brehm, N M; Combs, D K; Bauman, L M; Peters, J B; Undersander, D J

    1999-04-01

    Two studies were conducted to assess whether routine applications of near infrared reflectance spectroscopy could predict the effects of silage proteolysis on ruminal crude protein (CP) degradation of legume and grass silages. A preliminary study was conducted to assess the effect of laboratory drying method on ruminal CP degradation of silages. Thirty legume and grass silages were freeze-, oven-, or microwave-dried and incubated in situ in the ventral rumen of three ruminally cannulated cows for 24 h. Freeze-drying was considered least likely to alter ruminal CP degradation of the silages; therefore, oven- and microwave-drying were compared using first-order regression with freeze-drying. Oven-drying for 48 h at 55 degrees C compared favorably (R2 = 0.84) with freeze-drying. Microwave-drying resulted in a large bias (2.84 g/10(-1) kg of CP) and was poorly related (R2 = 0.48) to freeze-drying. In a second study, alfalfa and timothy were cut at three maturities and allowed to wilt for 0, 10, 24, 32, 48, and 54 h. Forages were ensiled in triplicate cylindrical mini silos and allowed to ferment for 120 d. After fermentation, silages were oven-dried, ground, and scanned on a near-infrared reflectance spectrophotometer. Duplicate, dried, 2-mm ground silage samples were incubated in the ventral rumen of three ruminally cannulated cows for 24 h. Forage species, maturity, and wilting time significantly affected 24-h ruminal CP degradation of the silages. Near-infrared reflectance spectroscopy accurately predicted (R2 = 0.91) 24-h ruminal CP degradation of silages. Data suggest near-infrared reflectance spectroscopy can accurately assess the effects of forage species, maturity, and wilting time (proteolysis) on 24-h ruminal CP degradation of legume and grass silages.

  17. Ruminal degradation kinetics of protein foods by in vitro gas production technique

    Directory of Open Access Journals (Sweden)

    Ivone Yurika Mizubuti

    2014-02-01

    Full Text Available Chemical analysis of carbohydrates and nitrogen fractions, as well as, determination their carbohydrates digestion rates in soyben meal (SM, crambe meal (CM, radish meal (RM, wet brewery residue (WBR and dehydrated silkworm chrysalis (SCD were accomplished. The kinetics parameters of non-fibrous carbohydrates (NFC and B2 fraction were estimated using cumulative gas production technique. Among the foods studied there was considerable variation in chemical composition. The crambe meal was the only food that did not present synchronism between carbohydrate and nitrogen fractions. In this food there was predominance of A+B1 carbohydrates fractions and B1+B2 nitrogen compounds fraction, and for the other predominated B2 carbohydrate fraction and B1+ B2 nitrogen compounds fraction. There were differences among the digestive kinetic parameters for all foods. The greater participation in gas production due to non-fibrous carbohydrates was found in the crambe meal and oilseed radish meal. The fermentation of fibrous carbohydrates provided higher gas volume in the wet brewery residue and in the soybean meal, however, the soybean meal was food with higher total gas volume. Non fibrous carbohydrates degradation rates of wet brewery residue and dehydrated silkworm chrysalis were far below the limits of degradation of this fraction. Due to the parameters obtained by the cumulative gas production, the soybean meal was the best food, however, all others have potential for use in animal nutrition. The cumulative gas production technique allows the estimative of degradation rates and provides further information about the ruminal fermentation kinetics of foods.

  18. 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...

  19. Protein preservation and ruminal degradation of ensiled forage treated with heat, formic acid, ammonia, or microbial inoculant.

    Science.gov (United States)

    Polan, C E; Stieve, D E; Garrett, J L

    1998-03-01

    The objectives of this study were to determine whether treatment of forage with heat would reduce proteolysis during subsequent fermentation. In Experiment 1, direct-cut barley forage and alfalfa were untreated, microwaved, or steamed and then ensiled in laboratory silos as wilted forages. Silages of microwaved or steamed forage showed a marked increase in N bound to neutral detergent fiber and in the recovery of protein; however, alfalfa silages also had high pH values and concentrations of butyric acid. In Experiment 2, steam heating was compared with formic acid and NH3 treatments for the prevention of proteolysis in alfalfa silages. Silage of steamed alfalfa had a greater amount of N bound to neutral detergent fiber and greater recovery of protein than did control silage or silages of forage treated with formic acid or NH3. Silage of steamed forage had lower pH values than did silages of wilted, direct-cut, or control forage. Microbial innoculant added to steamed forage increased the recovery of protein. Silage of steamed forage had less aerobic stability than did silage of direct-cut forage. Ruminal degradability of crude protein (CP) and organic matter of silage from both experiments was evaluated. Degradability of CP was 8 to 26 percentage units lower in silages of microwaved or steamed forage in Experiment 1 than in silage of unheated forage because of slower degradation rates, but all had similar undegraded CP after incubation for 72 h. In Experiment 2, wilting, steam, formic acid, and NH3 treatments affected CP degradability similarly, but CP degradability was decreased when compared with silage of direct-cut forage without treatment.

  20. Protein digestion in ruminants

    African Journals Online (AJOL)

    Animal Nutrition, Animal and Dairy Science Research Institute, Irene, 1675Republic of South Africa. Although the protein requirement of domestic ruminants may be calculated from a simple one-compartment model, this approach ignores factors such as microbial fermentation in the rumen and the non-equality of feed.

  1. Protein digestion in ruminants

    African Journals Online (AJOL)

    acids absorbed into the circulation of the animal. Ideally, therefore, the biological value of a feed protein should be determined from the amount and type of amino acid appearing in the portal circulation of the animal, and not simplythe dissappearance of amino acids from the tract. Ruminant digestion may be more easily ...

  2. Meeting the protein requirements of ruminant livestock | Beever ...

    African Journals Online (AJOL)

    Ruminant animals acquire their amino acids from the small intestinal digestion of ruminally derived microbial protein and dietary protein which has escaped ruminal degradation. The UK metabolizable protein system provides a framework with which the net absorption of amino acids from the small intestine is computed in ...

  3. Effects of electron beam irradiation on chemical composition, antinutritional factors, ruminal degradation and in vitro protein digestibility of canola meal

    Energy Technology Data Exchange (ETDEWEB)

    Taghinejad-Roudbaneh, M., E-mail: mtaghinejad@iaut.ac.i [Department of Animal Science, Faculty of Agriculture, Islamic Azad University, Tabriz Branch, P.O. Box 51589, Tabriz (Iran, Islamic Republic of); Ebrahimi, S.R. [Department of Animal Science, Faculty of Agriculture, Shahr-e-Qods Branch, Islamic Azad University, P.O. Box 37515-374, Shahr-e-Qods (Iran, Islamic Republic of); Azizi, S. [Department of Clinical Sciences, Faculty of Veterinary Medicine, Urmia University, P.O. Box 57155-1177, Urmia (Iran, Islamic Republic of); Shawrang, P. [Nuclear Science and Technology Research Institute, Agricultural, Medical and Industrial Research School, Atomic Energy Organization of Iran, P.O. Box 31485-498, Karaj (Iran, Islamic Republic of)

    2010-12-15

    The aim of the present study was to determine the impact of electron beam (EB) irradiation at doses of 15, 30 and 45 kGy on the nutritional value of canola meal. The phytic acid and total glucosinolate content of EB-irradiated canola meal decreased as irradiation doses increased (P<0.01). From in situ results, irradiation of canola meal at doses of 45 kGy decreased (P<0.05) the effective degradibility of crude protein (CP) by 14%, compared with an untreated sample. In vitro CP digestibility of EB-irradiated canola meal at doses of 15 and 30 kGy was improved (P<0.05). Electrophoresis results showed that napin and cruciferin sub-units of 30 and 45 kGy EB-irradiated canola meal were more resistant to degradation, compared with an untreated sample. Electron beam irradiation was effective in protecting CP from ruminal degradation and reducing antinutritional factors of irradiated canola meal.

  4. 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

  5. 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.

  6. Effects of electron beam irradiation on chemical composition, antinutritional factors, ruminal degradation and in vitro protein digestibility of canola meal

    Science.gov (United States)

    Taghinejad-Roudbaneh, M.; Ebrahimi, S. R.; Azizi, S.; Shawrang, P.

    2010-12-01

    The aim of the present study was to determine the impact of electron beam (EB) irradiation at doses of 15, 30 and 45 kGy on the nutritional value of canola meal. The phytic acid and total glucosinolate content of EB-irradiated canola meal decreased as irradiation doses increased ( Pcanola meal at doses of 45 kGy decreased ( Pcanola meal at doses of 15 and 30 kGy was improved ( Pcanola meal were more resistant to degradation, compared with an untreated sample. Electron beam irradiation was effective in protecting CP from ruminal degradation and reducing antinutritional factors of irradiated canola meal.

  7. Influence of two hop (Humulus lupulus L.) varieties on in vitro dry matter and crude protein degradability and digestibility in ruminants.

    Science.gov (United States)

    Lavrenčič, Andrej; Levart, Alenka; Košir, Iztok Jože; Cerenak, Andreja

    2014-04-01

    Hop cones contain several antimicrobial substances. The aim of the study was to determine the effects of two hop varieties, Aurora and Dana, on substrate (diet for a dairy cow, producing 30 kg milk daily) in vitro dry matter (DM) and crude protein (CP) degradability and digestibility. In the in vitro trial freshly ground hops were added to the buffered rumen fluid in concentrations simulating the cow's daily intake of 50, 100 and 200 g of hops. Increasing the concentration of hops decreased (P digestibility. The difference between CP digestibility and degradability represents an estimate of the amount of rumen 'bypass' protein which increased with increasing concentration of hops from 172 to 454 g kg(-1). Decreased DM and CP degradability and increased amount of rumen 'bypass' protein could lower the amounts of protein required by high-producing ruminant animals. However, this supposition needs a validation with in vivo trials. © 2013 Society of Chemical Industry.

  8. In situ ruminal degradation of amino acids and in vitro protein digestibility of undegraded CP of dried distillers' grains with solubles from European ethanol plants.

    Science.gov (United States)

    Westreicher-Kristen, E; Steingass, H; Rodehutscord, M

    2013-12-01

    The objectives of this study were to compare the in situ ruminal degradation of CP and amino acids (AAs) of dried distillers' grains with solubles (DDGS), and to estimate intestinal digestibility (ID) of undegradable crude protein (UDP) with the in vitro pepsin-pancreatin solubility of CP (PPS), using either DDGS samples (DDGS-s) or DDGS residues (DDGS-r) obtained after 16 h ruminal incubation. Thirteen samples originating from wheat, corn, barley and blends were studied. Lysine and methionine content of DDGS-s varied from 1.4 to 4.0 and 1.3 to 2.0 g/16 g N, respectively. The milk protein score (MPS) of DDGS-s was low and ranged from 0.36 to 0.51, and lysine and isoleucine were estimated to be the most limiting AAs in DDGS-s and DDGS-r. DDGS-r contained slightly more essential AAs (EAAs) than did the DDGS-s. Rumen degradation after 16 h varied from 44% to 94% for CP, from 39% to 90% for lysine and from 35% to 92% for methionine. Linear regressions showed that the ruminal degradation of individual AAs can be predicted from CP degradation. The PPS of DDGS-s was higher than that of DDGS-r and it varied from 70% to 89% and from 47% to 81%, respectively. There was no significant correlation between the PPS of DDGS-s and PPS of DDGS-r (R 2=0.31). The estimated intestinally absorbable dietary protein (IADP) averaged 21%. Moderate correlation was found between the crude fibre (CF) content and PPS of DDGS-r (R 2=0.43). This study suggests an overestimation of the contribution of UDP of DDGS to digestible protein supply in the duodenum in some currently used protein evaluation systems. More research is required and recommended to assess the intestinal digestibility of AAs from DDGS.

  9. 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

  10. Meeting the protein requirements of ruminant livestock D.E. Beever

    African Journals Online (AJOL)

    Ruminant animals acquire their amino acids from the small intestinal digestion of ruminally derived microbial protein and dietary protein which has escaped ruminal degradation. The UK metabolizable protein system provides a fiamework with which the net absorption of amino acids from the small intestine is computed in ...

  11. Degradabilidade ruminal da matéria seca e da proteína bruta de folhas e folíolos de forrageiras tropicais Ruminal degradabilities of dry matter and crude protein of tropical forages

    Directory of Open Access Journals (Sweden)

    Cristina Mattos Veloso

    2006-04-01

    Full Text Available Foram determinadas as degradabilidades in situ da MS e PB dos folíolos de três leguminosas tropicais leucena (Leucaena leucocephala (Lam. de Wit, guandu (Cajanus cajan (L. Millsp. e soja perene (Neonotonia wightii e de folhas de outras duas forrageiras tropicais rami (Boehmeria nivea (L. Gaudich e mandioca (Manihot esculenta Crantz. Amostras de 3 g (guandu, soja e rami ou 5 g (leucena e mandioca do material foram incubadas no rúmen de quatro novilhos por períodos de 3, 6, 12, 18, 24 e 48 horas. As degradabilidades efetivas da MS e da PB para a taxa de passagem de 5% hora foram elevadas (acima de 60%. As taxas de degradação, exceto a do guandu, também foram consideradas elevadas. As forrageiras apresentaram alta digestibilidade total da proteína (acima de 88%, exceto os folíolos de guandu (69%. O guandu foi a forrageira com menor potencial de degradação da proteína.In situ DM and CP degradabilities of leaflets of three tropical legumes: leucaena (Leucaena leucocephala (Lam. de Wit, pigeon pea (Cajanus cajan (L. Millsp., and glycine (Neonotonia wightii and of leaves of two other tropical forages: ramie (Boehmeria nivea (L. Gaudich and cassava (Manihot esculenta Crantz were determined in this trial. Three grams of pigeon pea, glycine, and ramie and 5 g of leucaena and cassava were incubated in the rumen of four steers for 3, 6, 12, 18, 24, and 48 hours. Effective degradabilities of DM and CP assuming a passage rate of 5%/hour were greater than 60%. Degradation rates, except for pigeon pea, were also high in this study. Forages had high total digestibility of protein (greater than 88%, except for pigeon pea leaflets that was 69%. Pigeon pea was the forage with the lowest potential for ruminal protein degradation.

  12. Ruminal degradability and intestinal digestibility of individual amino acids in mixed diets with different crude protein levels measured by the modified in vitro three-step and mobile nylon bag technique.

    Science.gov (United States)

    Gao, Wei; Zhang, Bowen; Lv, Bo; Liu, Chenli; Chen, Daofu

    2016-04-01

    The ruminal degradability and intestinal digestibility of dry matter (DM), crude protein (CP) and amino acids (AA) in three total mixed rations with different CP levels were estimated using the modified in vitro three-step procedure (TSP) and mobile nylon bag (MNB) technique on growing lambs. The ruminal effective degradability of DM and CP did not respond with increasing dietary CP level. However, the intestinal digestibility of DM was significantly increased with increasing dietary CP level estimated by TSP (P digestibility coefficients of CP determined by TSP were lower than those of the MNB method. Histidine was extensively degraded by rumen micro-organisms, while tyrosine was the most anti-degradable AA among the samples. The ruminal AA degradability exhibited no significant differences except for threonine, tryptophan, alanine, aspartic acid and proline for the three diets. Similarly, only a few AAs (i.e. histidine, methionine, tryptophan, aspartic acid and cysteine in TSP; histidine, tryptophan, aspartic acid and serine in MNB) had significant differences in their intestinal digestibility; in addition, values of MNB were lower than that of the TSP method, indicating that intestinal digestibility of DM seems to be overestimated in TSP, while that of CP might be overestimated in the MNB method. © 2015 Japanese Society of Animal Science.

  13. Accounting for the effects of a ruminal nitrogen deficiency within the structure of the Cornell Net Carbohydrate and Protein System

    National Research Council Canada - National Science Library

    Tedeschi, L. O; Fox, D. G; Russell, J. B

    2000-01-01

    .... The Cornell Net Carbohydrate and Protein System (CNCPS) prediction of fiber digestion and microbial mass production from ruminally degraded carbohydrate has been adjusted to accommodate a ruminal N deficiency...

  14. Accounting for the effects of a ruminal nitrogen deficiency within the structure of the Cornell Net Carbonhydrate and Protein System

    National Research Council Canada - National Science Library

    L O Tedeschi; D G Fox; J B Russell

    2000-01-01

      The Cornell Net Carbohydrate and Protein System (CNCPS) prediction of fiber digestion and microbial mass production from ruminally degraded carbohydrate has been adjusted to accommodate a ruminal N deficiency...

  15. Effect of heat treatment and subsequent urea supplementation of sunflower meal on the in vitro ruminal degradability of crude protein content and its postruminal digestibility.

    Science.gov (United States)

    Veresegyházy, T; Kutas, F; Fekete, S; Csapó, J

    1989-01-01

    A three-phase laboratory procedure was used for predicting the degradability of the protein of extracted sunflower meal before (SFM) and after heat treatment (HSFM). The rumen fluid degraded 69.1% and 67.1% of the SFM and HSFM protein, respectively. The digestibility values of rumen undegraded protein (UDP) were 57.3% (SFM) and 57.0% (HSFM) with pepsin and 17.6% (SFM) and 15.9% (HSFM) with pancreatin. Urea supplementation practically did not alter the rumen degradability of HSFM protein, while the pepsin digestibility of UDP decreased to 47.2%. Four fractions (NH3, dissolved amino acids, oligopeptides and proteins) of rumen degradable crude protein (RDP) were also determined in vitro: 81 to 92% of the degraded crude protein was found in the fractions tested. Heat treatment reduced free NH3 content but did not alter the other three fractions. Urea supplementation decreased the quantity of NH3, peptides and proteins as well.

  16. RUMINAL DEGRADATION OF THE DRY MATTER, PROTEIN AND THE AMINOACIDS OF THE CORN AND CORN GLUTENS MEAL DEGRADAÇÃO RUMINAL DA MATÉRIA SECA, DA PROTEÍNA E DOS AMINOÁCIDOS DO MILHO E DE GERMENS DE MILHO

    Directory of Open Access Journals (Sweden)

    Juliana Borbari Dourado

    2007-09-01

    Full Text Available

    The present work was carried out at the Faculty of Agriculture and Animal Science, UNESP-Jaboticabal. Three ¾ Holstein x Zebu bovines fistulated on rumen, abomasum and intestine using a fixed cannula type T were used. Four types of feed: corn (M, 1% EE corn gluten meal (G1, 7% EE corn gluten meal and 10% EE corn gluten meal (G10 were tested.  All feed were also extruded: CornEx, G1Ex, G7Ex and G10EX. Ruminal incubation was done using the nylon bags technique, and degradability of dry matter, crude protein and disponibilization of amino acids were analyzed. Basic diet was composed of corn gluten meal, soybean meal and hay at roughage (70: concentrate (30 ratio. Ruminal degrading of dry matter feed M, G1, G7 and G10 presented the following values: 37.5%, 56.7%, 56.8% and 55.1% respectively.  Extruded feed: 52.3%, 68.9%; 69.0% and 61.6% for Mex, G1Ex, G7Ex and G10Ex. The corn was significantly inferior to others and extruded feed increased ruminal degradedly for all feed. The degradability of crude protein was 27.0%, 60.9%, 56.8% and 35.1% for M, G1, G7 and G10; 50.8, 52.2, 66.4 and 59.6 for MEx; G1Ex; G7Ex and G10Ex, respectively. Statistically the G1 feed was equal to M and both were inferior to G7 and G10. The extrusion process did not increase the degradability of crude protein of G1. The lysine presented high ruminal degradability for tested feed, showing values above 99%. The metionin presented values of degradability inferior for M (37.4% related to G1 (57.1%.  The treonine did not present differences between feed and extrusion. It can be concluded that the processing of corn to obtain by-products can increase the ruminal degradability of dry matter, protein and metionine, but depending on intensity of processing, the degradability of crude protein may not be altered when compared to corn. The extrusion increased the ruminal degradability of dry matter of corn and by-products.

    KEY-WORDS: Amino acids, degradability, extrusion

  17. Degradabilidade ruminal do feno de alguns alimentos volumosos para ruminantes Ruminal degradability of some roughage hays for ruminants feeding

    Directory of Open Access Journals (Sweden)

    G.G.P. Carvalho

    2006-08-01

    Full Text Available Avaliou-se a degradabilidade ruminal da matéria seca (MS, proteína bruta (PB, fibra em detergente neutro (FDN, da fibra em detergente ácido (FDA e hemicelulose dos fenos de capim-elefante (Pennisetum purpureum, palma (Opuntia ficus, guandu (Cajanus cajan e parte aérea da mandioca (Manihot esculenta utilizando três bovinos mestiços machos, castrados, canulados no rúmen e mantidos em regime de pasto. Amostras de 4g de cada alimento foram incubadas em duplicata no rúmen dos animais, nos períodos de 0, 6, 12, 24, 36, 48 e 72 horas. A degradabilidade potencial da PB dos fenos de capim-elefante e guandu foi semelhante, 83,9 e 81,2%, respectivamente. Os maiores valores foram observados para os fenos de palma (94,2% e parte aérea da mandioca (91,7%. A degradabilidade efetiva (DE foi obtida considerando as taxas de passagem de 2, 5 e 8%/hora. A maior DE observada para MS (60,5%, PB (81,1%, FDN (21,6%, FDA (27,9% e HEM (58,0%, na taxa de passagem de 5%/h, ocorreu com o feno de palma.The ruminal degradability of dry matter (DM, crude protein (CP, neutral detergent fiber (NDF, acid detergent fiber (ADF and hemicellulose (HEM of elephantgrass (Pennisetum purpureum, forage cactus (Opuntia ficus, pigeon pea (Cajanus cajan and cassava foliage (Manihot esculenta hays was evaluated using three cannulated crossbred steers, kept on pasture. Samples of four grams of each hay were incubated in the rumen for 0, 6, 12, 24, 36, 48 and 72 hours. The CP potential degradability (PD for elephantgrass and pigeon pea hays was similar, 83.9 and 81.2%, respectively. Higher values were observed either for forage cactus (94.2% or cassava foliage (91.7% hays. The effective degradability (ED was obtained considering the passage rates of 2, 5 and 8%/hour. The forage cactus hay, at a passage rate of 5%/h, showed the highest ED for DM (60.5%, CP (81.1%, NDF (21.6%, ADF (27.9% and HEM (58.0%.

  18. Comparison of analyses to predict ruminal fibre degradability and ...

    African Journals Online (AJOL)

    The objective of this study was to compare the ruminal degradability of neutral detergent fibre (NDF) and indigestible NDF (INDF) between silages (n = 24) that originated from three different temperate grass species, i.e. Dactylis glomerata L., Festuca arundinacea L. and hybrid, Felina – Lolium multiflorum L. × Festuca ...

  19. In Vitro Degradation of Melamine by Ruminal Microorganisms ...

    African Journals Online (AJOL)

    An in vitro study was conducted to determine the extent of melamine degradation in rumen liquor. Rumen liquor was collected from two ruminally cannulated Holstein cows on four separate dates, one week apart. Erlenmeyer flasks (250 mL) were prepared for incubation by adding 1000 mg of a dairy feed substrate, 100 mg ...

  20. Productivity, chemical composition and ruminal degradability of ...

    African Journals Online (AJOL)

    ShoeskY

    2016-11-19

    Nov 19, 2016 ... Information on the relationship between the quality and degradability of Napier grass at various stages of maturity is limited and is ... experimental period were extracted from Agricultural Research Council – Institute for Soil, Climate and Water. (ARC-ISCW) station ...... Region of Colombia. II. Trace Minerals.

  1. In vitro degradation of melamine by ruminal microorganisms

    African Journals Online (AJOL)

    Tanja&Jana

    2015-05-17

    May 17, 2015 ... hydrolysis in the rumen was insufficient to promote maximum ruminal protein synthesis and concluded that melamine was not an ... ADF and NDF. Heat-stable alpha-amylase and sodium sulphite were used in the assay. The .... A comparison between nitrogen retention from urea, biuret, triuret and cyanuric ...

  2. Evaluation of a Commercial ELISA for Detection of Ruminant Processed Animal Proteins in Non-Ruminant Processed Animal Proteins

    NARCIS (Netherlands)

    Bremer, M.G.E.G.; Margry, R.J.C.F.; Vaessen, J.C.H.; Doremalen, van A.M.H.; Palen, van der J.G.P.; Kaathoven, van R.G.C.; Kemmers-Voncken, A.E.M.; Raamsdonk, van L.W.D.

    2013-01-01

    Due to a growing aquaculture industry, demand for high-quality proteins for aquatic feeds is increasing. Non-ruminant processed animal proteins (PAPs) have shown great potential for this purpose. Safe reintroduction of non-ruminant PAPs in aqua feed requires methods that can discriminate ruminant

  3. Ruminal Bacterial Degradation of Benzo(b)-thien-4-yl Methylcarbamate (Mobam) and Effect of Mobam on Ruminal Bacteria

    Science.gov (United States)

    Williams, Phletus P.; Stolzenberg, Rosa L.

    1972-01-01

    Mixtures of ruminal bacteria degraded benzo(b)thien-4-yl methylcarbamate (Mobam) to 4-hydroxybenzothiophene, CO2, and polar product(s). The metabolite, 4-hydroxybenzothiophene, was identified (after acetylation) by comparative infrared and mass spectrometry with an authentic sample. Carbon dioxide and polar product(s) were produced by degradation of the methylcarbamate moiety. Ten previously characterized strains of ruminal bacteria with diverse physiological capabilities did not degrade Mobam. However, three tributyrin-hydrolyzing strains were isolated that did degrade Mobam. Mobam inhibited growth of two of ten strains isolated on Mobam-free glycerol-tributyrin enrichment medium. One of these strains was also sensitive to 2-carbomethoxy-propene-2yl dimethyl phosphate (Phosdrin). Mobam prevented some ruminal bacteria from producing zones of hydrolysis in tributyrin emulsion media and inhibited some ruminal bacteria from degrading 1-naphthyl acetate and fluorescein-3′,6′-diacetate. PMID:4553142

  4. Degradação ruminal da matéria seca e proteína bruta, de alimentos concentrados utilizados como suplementos para novilhos Ruminally degradation of the dry matter and crude protein, of concentrade feeds used as supplementation in diet for young bulls

    Directory of Open Access Journals (Sweden)

    Rafael Henrique de Tonissi e Buschinelli de Goes

    2004-02-01

    Full Text Available Com o objetivo de avaliar a degradabilidade ruminal da matéria seca (MS e da proteína bruta (PB de diferentes alimentos concentrados utilizados na alimentação de bovinos, pela técnica in situ, foram utilizados três novilhos Nelores, inteiros, com 24 meses e peso aproximado de 350 Kg, fistulados no rúmen. As amostras referentes aos alimentos foram pesadas e colocadas no interior de sacos de náilon (6,5 x 4,0 cm de área livre, com porosidade conhecida de 50 micras, na quantidade de 1,0 grama de MS; esses foram incubados em ordem decrescente no rúmen nos tempos de 48, 36, 24, 12, 6, 3 e 0 horas. Foram avaliados seis alimentos: glúten de milho, polpa cítrica, farelo de soja, farelo de trigo, milho moído e o farelo de amendoim. Os dados foram ajustados por regressão não-linear, segundo o modelo p = a + b(1-e-ct. As degradabilidades efetivas (DE da matéria seca dos alimentos foram, respectivamente, de 29,3; 76,6; 71,3; 60,5; 59,6; e 60,8, para a taxa de passagem de 5%/h, e da proteína bruta foram de 8,5; 56,1; 57,4; 67,9; 30,1; e 85,2, para a mesma taxa de passagem, respectivamente. A polpa cítrica apresentou alto valor para a degradabilidade efetiva de MS (76,6%; o milho, uma lenta degradação ruminal da PB, com uma taxa de degradação para a fração “b” de 2,9%; o glúten de milho apresentou uma fração “a” de 11,9% para a MS, sendo uma fonte de proteína de alto escape ruminal, apresentou DE de 8,5%; o farelo de amendoim apresentou valor, para a fração “b”, de 67,3%, o valor nutritivo desse alimento é semelhante ao farelo de soja, mas teve a maior DE de PB superior 48% ao farelo de soja. O glúten de milho apresentou baixa degradação ruminal, enquanto a polpa cítrica e o milho tiveram uma lenta degradação ruminal, principalmente quando comparados ao farelo de amendoim.With the objective of evaluating the ruminal degradation of the dry matter and the crude protein of different concentrated feed used when

  5. In vitro ruminal fermentation and in situ ruminal degradation of tamarind kernel powder extract residue in wethers.

    Science.gov (United States)

    Wang, Lin; Oishi, Kazato; Sato, Yoshiaki; Nakanishi, Takashi; Hirooka, Hiroyuki; Takahashi, Kei; Kumagai, Hajime

    2017-07-01

    We examined the in vitro fermentation characteristics and in situ dry matter (DM) and crude protein (CP) degradability of tamarind kernel powder extract residue (TKPER), a by-product of polysaccharides thickener processing. Two types of TKPER (I and II), of which the CP and neutral detergent fiber organic matter basis contents (%) were 41.4 and 42.0 and 1.4 and 0.5, respectively, were compared with dry heat soybean (SB), soybean meal (SBM) and dry soybean curd residue (SBCR). The TKPERs had significantly lower in vitro gas production compared to the other products at each observation time (P < 0.05). The in vitro DM and CP digestibility (%) of TKPER I and II were 67.7 and 64.9, and 64.5 and 58.0, respectively, significantly lower than those of SB and SBM (P < 0.05). We used four wethers (55.6 ± 4.6 kg) with ruminal cannulas to investigate the in situ degradability of TKPER I, SB, SBM and SBCR. At the ruminal outflow rate of 0.05/h, the effective degradability (%) values of the DM and CP of TKPER I were 64.1 and 49.7, respectively, which were similar to those of SBM. In conclusion, TKPER had high CP and exhibited in situ degradability similar to that of SBM, suggesting that TKPER could be used as a protein source feed. © 2016 Japanese Society of Animal Science.

  6. Diversity, abundance and characterization of ruminal cysteine phytases suggest their important role in phytate degradation.

    Science.gov (United States)

    Huang, Huoqing; Zhang, Rui; Fu, Dawei; Luo, Jianjie; Li, Zhongyuan; Luo, Huiying; Shi, Pengjun; Yang, Peilong; Diao, Qiyu; Yao, Bin

    2011-03-01

    A novel class of cysteine phytase showing ability to degrade phytate has recently been isolated from rumen bacteria. To expand our knowledge of this enzyme class, a total of 101 distinct cysteine phytase gene fragments were identified from the ruminal genomic DNA of Bore goats and Holstein cows, and most of them shared low identities (cysteine phytase genes of goat rumens were cloned and their protein products were characterized. Four findings were revealed based on our results. (i) Compared with soil and water environment, where β-propeller phytase is the most important phytate-degrading enzyme, cysteine phytase is the major phytate-degrading enzyme in the anaerobic ruminal environment. (ii) Cysteine phytase fragments in the rumen contents of goat and cow have the same diversity profile, although most of the sequences and their abundance differ in the two species. (iii) Each species has their respective high-abundance genes, which may play major roles for phytate degradation. (iv) Compared with previously reported cysteine phytases that have pH optimum at 4.5, the pH optima of the two most abundant secreted goat cysteine phytases are 6.5 and 6.0, which are within the pH range found in the rumens. This study provides valuable information about the diversity, abundance and enzymatic properties of the ruminal cysteine phytases and emphasizes the important role(s) of these cysteine phytases probably in the terrestrial cycle of phosphorus. © 2010 Society for Applied Microbiology and Blackwell Publishing Ltd.

  7. Degradabilidade ruminal In Situ da matéria seca e proteína bruta das silagens de milho e sorgo e de alguns alimentos concentrados Ruminal in situ degradability of dry matter and crude protein of corn and sorghum silages and some concentrate feeds

    Directory of Open Access Journals (Sweden)

    Adriana de Souza Martins

    1999-01-01

    Full Text Available A degradação ruminal da matéria seca (MS e a da proteína bruta (PB de alguns alimentos foram deterninadas pela técnica in situ, usando-se três vacas Holandesa. Os alimentos concentrados foram moídos em peneira de 2 mm e os volumosos e a raspa de mandioca, em peneira de 5 mm, e incubados em sacos de náilon (53µ. A degradabilidade efetiva (DE da MS (5%/h de silagem de milho, silagem de sorgo, milho, sorgo, farelo de trigo, triticale, raspa de mandioca, polpa cítrica, farelo de soja, farelo de algodão e farinha de carne e ossos foi, respectivamente: 54,8; 46,0; 37,6; 54,3; 80,4; 88,7; 75,0, 67,5; 74,1; 65,9; e 39,4%. As degradabilidades efetivas da PB, 70,4; 67,9; 40,2; 42,7; 86,5; 89,3; 71,7; 62,1; 57,3; 81,4; e 47,5%, respectivamente. Foi realizado agrupamento dos alimentos quanto à dinâmica de degradação ruminal da MS e PB foram por meio do método de Tocher. A fração potencialmente degradável e a sua taxa de degradação foram as variáveis de maior poder discriminatório na formação dos grupos.The ruminal dry matter (DM and crude protein (CP degradability of some feeds were evaluated by the in situ technique, using three Holstein cows. The concentrate feeds were ground through a 2 mm screen and the forage and the cassava root, in 5 mm screen, and incubated in nylon bag (53µ. The effective degradability (ED of DM (5%/h for corn silage, sorghum silage, corn, sorghum, wheat bran, triticale, cassava scraping, citrus pulp, soybean meal, cottonseed meal and meat and bone meal was: 54.8, 46.0, 37.6, 54.3, 80.4, 88.7, 75.0, 67.5, 74.1, 65.9, and 39.4%, respectively. The effective degradabilities CP were: 70.4, 67.9, 40.2, 42.7, 86.5, 89.3, 71.7, 62.1, 57.3, 81.4, and 47.5%, respectively. The feed group was made according to ruminal DM and CP degradation dynamics, by means of Tocher method. The potentially degradable fraction and its degradation rate were the variables of the highest discrimination power for the group formation.

  8. Subacute ruminal acidosis challenge changed in situ degradability of feedstuffs in dairy goats.

    Science.gov (United States)

    Li, Fei; Cao, Yangchun; Liu, Nannan; Yang, Xinjian; Yao, Junhu; Yan, Dabing

    2014-01-01

    This study investigated the effects of wheat-induced subacute ruminal acidosis (SARA) on rumen bacterial populations and in situ degradabilities of NDF, starch, and crude protein of feeds. Four multiparous dairy goats (BW=60±3.3kg) fitted with ruminal cannulas were assigned to a 2×2 crossover design (28-d treatment periods separated by a 7-d washout interval). The treatment diets consisted of 2 levels of cracked wheat: 0 (control, corn based concentrate) and 35% (diet-induced SARA, wheat-based concentrate), with a constant forage- (45% alfalfa hay and 5% corn silage of DM) to-concentrate (50% of DM) ratio. Results indicate that diets with a 35% wheat decreased ruminal pH (6.21 vs. 5.98) and increased the duration (1.13 vs. 4.72h/d) and area (0.12 vs. 0.78 pH × h/d) of ruminal pH below 5.6 and induced SARA. The SARA increased ruminal total volatile fatty acid concentration, from 105.0 to 123.8mM, and decreased the acetate molar proportion (62.8 vs. 56.6mol/100mol) and the acetate-to-propionate ratio (3.5 vs. 2.8). Compared with the control group, SARA decreases the relative abundance of Fibrobacter succinogenes (-59.3%) and Ruminococcus flavefaciens (-68.4%), whereas it increased Succinimonas amylolytica (198.1%) and Ruminobacter amylophilus (125.2%). The SARA decreased 24- and 48-h dry matter (DM) and neutral detergent fiber (NDF) degradabilities of corn silage. The 48-h degradabilities of DM (51.0 vs. 48.2%) and NDF (40.3 vs. 36.0%) in alfalfa hay were not affected by SARA, but the SARA tended to reduce the 24-h DM (49.6 vs. 46.3%) and NDF (37.8 vs. 33.2%) degradabilities. The effective ruminal degradabilities of DM and NDF in alfalfa hay and corn silage were reduced during SARA. In situ degradability parameters of DM and starch of wheat were not affected by SARA, but starch degradability of corn (9.5 vs. 13.3%/h) increased. The SARA reduced in situ 12-h degradabilities of DM and crude protein of soybean meal and extruded soybean without affecting the

  9. Fermentation, kinetics and ruminal degradation in sheep fed with carbohydrate sources associated with oil

    Directory of Open Access Journals (Sweden)

    Eliane da Silva Morgado

    2013-12-01

    Full Text Available The objective this work was to evaluate the degradation, fermentation and kinetics of passage rumen fluid en sheep fed diets containing different carbohydrate associated with the oil. Four rumen cannulated sheep were allotted in 4 x 4 latin square. The treatments consisted of diets with high neutral detergent soluble fiber and diets with high starch associated or not with 4,2% of oil. Incubation times were 3, 6, 12, 24, 48, 72, 96 and 120 hours and ruminal fluid was collected at 0, 2, 4, 6, 8, 10, 12, and 24 hours after feeding. Diets high neutral detergent fiber soluble resulted in greater degradation of dry matter, crude protein, neutral detergent fiber soluble and starch, and higher pH values, butyric acid production, dilution rate and ruminal recycling compared with diets high starch content. The inclusion of 4,2% oil the different carbohydrate sources had no influence on the fermentation kinetics and degradation of the fiber. High content of neutral detergent soluble fiber in the diet favors the ruminal fermentation compared to the high starch content.

  10. Proteína degradável no rúmen associada a fontes de amido de alta ou baixa degradabilidade: digestibilidade in vitro e desempenho de novilhos em crescimento Rumen degradable protein associated to starch sources of different ruminal degradability: in vitro digestibility and performance of growing steers

    Directory of Open Access Journals (Sweden)

    Saul Ferreira Caldas Neto

    2007-04-01

    Full Text Available Objetivou-se avaliar os efeitos de teores crescentes de proteína degradável no rúmen (PDR = 47, 52, 57 e 62% associados a fontes de amido de baixa (milho ou de alta (farinha de varredura de mandioca - FVM degradabilidade ruminal em rações com 50:50% volumoso:concentrado sobre a digestibilidade in vitro da MS (DIVMS e o ganho de peso de novilhos em crescimento. Para determinação da DIVMS, foi utilizada a técnica de dois estádios de Tilley & Terry (1963. As médias obtidas para a DIVMS foram submetidas às análises de variância e regressão em função do teor de PDR da ração. Para o ensaio de desempenho animal, foram utilizados 32 novilhos mestiços (Nelore x Red Angus, não-castrados (275 kg de PV, alojados em duplas, em delineamento experimental inteiramente casualizado. Não foi observado efeito do teor de PDR de dietas contendo milho sobre a DIVMS, indicando que não houve deficiência de nitrogênio e que, possivelmente, a digestibilidade da fonte de amido (milho foi o fator limitante. Observou-se efeito quadrático do teor de PDR sobre a DIVMS para as dietas contendo FVM, o que indica a ocorrência do efeito de sincronização entre a disponibilidade de energia e nitrogênio, permitindo aumento da atividade microbiana nos níveis mais elevados de PDR na ração e melhor utilização da energia proveniente da fonte de amido de alta degradabilidade ruminal. Não foi observado efeito dos diferentes teores de PDR e da fonte de amido de alta degradabilidade ruminal sobre o consumo de MS, o ganho de peso médio e a conversão alimentar dos animais.The objective of this study was to evaluate the effects of increasing levels of rumen-degradable protein (RDP = 47, 52, 57 and 62% associated to starch sources of low (corn and high (cassava by-product (CBP ruminal degradability on in vitro DM digestibility (IVDMD and average daily gain of growing steers. In vitro DM digestibility was determined by using the Tilley and Terry (1963 technique

  11. In situ ruminal degradability of dry matter, organic matter and crude protein of some concentrated supplements used in the feeding of bovines/ Degradabilidade ruminal “in situ” da matéria seca, matéria orgânica e proteína bruta de alguns suplementos concentrados usados na alimentação de bovinos

    Directory of Open Access Journals (Sweden)

    Marco Antônio da Rocha

    2005-06-01

    Full Text Available Holstein steers, with permanent ruminal cannula, 650 kg of liveweight, maintained on pasture, and receiving mineral mixture “ad libtum” were used. The present study had the objective of evaluating the “in situ” ruminal degradation of the dry matter (DM, crude protein (CP and organic matter (OM of 12 concentrated feed: whole sunflower seed (WSF, partially degreased sunflower (DSF, sunflower meal (SFM, sunflower pie pressed one time (T 1x, sunflower pie pressed two time (T 2x, for extracting of the oil, whole soybean seed (WS, partially degreased soybean (DS, soybean meal (SM, whole cottonseed (WC, cottonseed meal (CM, degreased corn germ (CG and a commercial concentrate with 36% of CP (CC. The feed were grounded in a mill equipped with a screen with sieves of 2 mm of diameter. Samples 5 g of DM from each one of the mentioned feed were incubated in nylon bags with pores of 50 micras of diameter, measuring 14 x 7 cm, for the times of 12; 20; and 33 h of ruminal fermentation. The effective degradability (ED of CP, for a passage rate of 5%/h varied from 62.08 to 95.93% for commercial concentrate and sunflower pie pressed two time for OM varied from 48.17 to 85.28% for soybean meal and whole cottonseed. The CC and CG presented the smallest ED of CP, however only the CC have larger protein escapes to be digested in the intestines, because CG presents little PB in its constitution.Foram utilizados quatro bovinos da raça holandesa, machos castrados, fistulados no rúmen, pesando em média 650kg, mantidos em pastejo e recebendo mistura mineral “ad libtum”. O presente trabalho teve como objetivo avaliar a degradação ruminal “in situ” da matéria seca (MS, proteína bruta (PB e matéria orgânica (MO de 12 alimentos concentrados: grão de girassol integral (GI, grão de girassol parcialmente desengordurado (GD, farelo de girassol (FG, torta de girassol com uma passagem pela prensa (T 1x, torta de girassol com duas passagens pela prensa (T

  12. Proteins Profile in Milk from Three Species of Ruminants

    National Research Council Canada - National Science Library

    Anamaria COZMA; Sanda ANDREI; Doina MIERE; Lorena FILIP; Felicia LOGHIN

    2011-01-01

    .... In this research, total proteins, caseins and whey proteins respectively, were dosed in milk from three species of ruminants-cow, goat and sheep, using a very sensitive method, Bradford photometric method...

  13. 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

  14. Prediction of CP and starch concentrations in ruminal in situ studies and ruminal degradation of cereal grains using NIRS.

    Science.gov (United States)

    Krieg, J; Koenzen, E; Seifried, N; Steingass, H; Schenkel, H; Rodehutscord, M

    2017-08-03

    Ruminal in situ incubations are widely used to assess the nutritional value of feedstuffs for ruminants. In in situ methods, feed samples are ruminally incubated in indigestible bags over a predefined timespan and the disappearance of nutrients from the bags is recorded. To describe the degradation of specific nutrients, information on the concentration of feed samples and undegraded feed after in situ incubation ('bag residues') is needed. For cereal and pea grains, CP and starch (ST) analyses are of interest. The numerous analyses of residues following ruminal incubation contribute greatly to the substantial investments in labour and money, and faster methods would be beneficial. Therefore, calibrations were developed to estimate CP and ST concentrations in grains and bag residues following in situ incubations by using their near-infrared spectra recorded from 680 to 2500 nm. The samples comprised rye, triticale, barley, wheat, and maize grains (20 genotypes each), and 15 durum wheat and 13 pea grains. In addition, residues after ruminal incubation were included (at least from four samples per species for various incubation times). To establish CP and ST calibrations, 620 and 610 samples (grains and bag residues after incubation, respectively) were chemically analysed for their CP and ST concentration. Calibrations using wavelengths from 1250 to 2450 nm and the first derivative of the spectra produced the best results (R 2 Validation=0.99 for CP and ST; standard error of prediction=0.47 and 2.10% DM for CP and ST, respectively). Hence, CP and ST concentration in cereal grains and peas and their bag residues could be predicted with high precision by NIRS for use in in situ studies. No differences were found between the effective ruminal degradation calculated from NIRS estimations and those calculated from chemical analyses (P>0.70). Calibrations were also calculated to predict ruminal degradation kinetics of cereal grains from the spectra of ground grains

  15. Silage fermentation and ruminal degradation of stylo prepared with lactic acid bacteria and cellulase.

    Science.gov (United States)

    Li, Mao; Zhou, Hanlin; Zi, Xuejuan; Cai, Yimin

    2017-10-01

    In order to improve the silage fermentation of stylo (Stylosanthes guianensis) in tropical areas, stylo silages were prepared with commercial additives Lactobacillus plantarum Chikuso-1 (CH1), L. rhamnasus Snow Lact L (SN), Acremonium cellulase (CE) and their combination as SN+CE or CH1 + CE, and the fermentation quality, chemical composition and ruminal degradation of these silages were studied. Stylo silages treated with lactic acid bacteria (LAB) or cellulase, the pH value and NH 3 -N ⁄ total-N were significantly (P fermentation and ruminal degradation than SN+CE treatment. The results confirmed that LAB or LAB plus cellulase treatment could improve the fermentation quality, chemical composition and ruminal degradation of stylo silage. Moreover, the combined treatment with LAB and cellulase may have beneficial synergistic effects on ruminal degradation. © 2017 Japanese Society of Animal Science.

  16. Effects of pressure toasting on in situ degradability and intestinal protein and protein-free organic matter digestibility of rapeseed

    NARCIS (Netherlands)

    Azarfar, A.; Ferreira, C.; Goelema, J.O.; Poel, van der A.F.B.

    2008-01-01

    BACKGROUND: Rapeseed is a protein supplement that contains up to 40% crude protein (CP) on a dry matter (DM) basis, but a large part of its protein can be easily degraded in the rumen. Therefore, before inclusion in ruminant's diet, the extent of its protein degradation in the rumen must be reduced

  17. In situ ruminal degradation of phytic acid in formaldehyde treated rice bran

    NARCIS (Netherlands)

    Martin-Tereso, J.; Gonzalez, A.; Laar, van H.; Burbara, C.; Pedrosa, M.; Mulder, K.; Hartog, den L.A.; Verstegen, M.W.A.

    2009-01-01

    Rice bran has a very high content of phytic acid (IP6), which is a nutritional antagonist of Ca. Microbial phytase degrades IP6, but ruminal degradation of nutrients can be reduced by formaldehyde treatment. Milk fever in dairy cows can be prevented by reducing available dietary Ca to stimulate Ca

  18. Fractionation of carbohydrate and protein content of some forage feeds of ruminants for nutritive evaluation

    Directory of Open Access Journals (Sweden)

    Lalatendu Keshary Das

    2015-02-01

    Full Text Available Aim: To evaluate some forage feeds of ruminants in terms of their carbohydrate (CHO and protein fractions using Cornell Net Carbohydrate and Protein System (CNCPS. Materials and Methods: Eleven ruminant feeds (six green fodders - maize, oat, sorghum, bajra, cowpea, berseem and five range herbages - para grass, guinea grass, hedge lucerne, setaria grass and hybrid napier were selected for this study. Each feed was chemically analyzed for proximate principles (dry matter, crude protein [CP], ether extract, organic matter and ash, fiber fractions (neutral detergent fiber, acid detergent fiber, acid detergent lignin, cellulose and hemicellulose, primary CHO fractions (CHO, non-structural CHO, structural CHO and starch and primary protein fractions (neutral detergent insoluble CP, acid detergent insoluble CP, non-protein nitrogen and soluble protein. The results were fitted to the equations of CNCPS to arrive at various CHO (CA - fast degrading, CB1 - intermediate degrading, CB2 - slow degrading and CC - nondegrading or unavailable and protein (PA - instantaneously degrading, PB1 - fast degrading, PB2 - intermediate degrading, PB3 - slow degrading and PC - non-degrading or unavailable fractions of test feeds. Results: Among green fodders, cowpea and berseem had higher CA content while except hedge lucerne all range herbages had lower CA values. CB1 content of all feeds was low but similar. All feeds except cowpea, berseem, and hedge lucerne contained higher CB2 values. Oat among green fodders and hybrid napier among range herbages had lower CC fraction. Feeds such as bajra, cowpea, berseem and the setaria grass contained lower PA fraction. All green fodders had higher PB1 content except maize and cowpea while all range herbages had lower PB1 values except hedge lucerne. Para grass and hybrid napier contained exceptionally low PB2 fraction among all feeds. Low PC contents were reported in oat and berseem fodders. Conclusion: Based on our findings, it

  19. Protein degradation systems in platelets.

    Science.gov (United States)

    Kraemer, B F; Weyrich, A S; Lindemann, S

    2013-11-01

    Protein synthesis and degradation are essential processes that allow cells to survive and adapt to their surrounding milieu. In nucleated cells, the degradation and/or cleavage of proteins is required to eliminate aberrant proteins. Cells also degrade proteins as a mechanism for cell signalling and complex cellular functions. Although the last decade has convincingly shown that platelets synthesise proteins, the roles of protein degradation in these anucleate cytoplasts are less clear. Here we review what is known about protein degradation in platelets placing particular emphasis on the proteasome and the cysteine protease calpain.

  20. Ruminal, Intestinal, and Total Digestibilities of Nutrients in Cows Fed Diets High in Fat and Undegradable Protein

    DEFF Research Database (Denmark)

    Palmquist, D.L.; Weisbjerg, Martin Riis; Hvelplund, Torben

    1993-01-01

    in a 2×2 factorial design in a 4×4 Latin square experiment. Ruminal degradabilities were 14.9 and 18.6%, and intestinal digestibilities were 98.9 and 68.3%, respectively, for CP in blood meal and feather meal. Treatment effects on ruminal digestibilities were small. Protein supplementation increased......To study relationships of high undegradable intake protein and dietary fat on intestinal AA supply, the ruminal, intestinal, and total digestibilities of diets with or without added fat (5% of DM) and animal protein (blood meal: hydrolyzed feather meal, 1:1; 8% of DM) were examined with four cows...... with cows cannulated 100-cm distal to the pylorus, but only when cows were fed protein-supplemented diets; the estimates from those diets caused calculated microbial protein efficiency to exceed theoretical values. We postulated that blood meal and feather meal segregated near the pylorus, yielding high...

  1. The evaluation of metabolizable protein content of some indigenous feedstuffs used in ruminant nutrition

    Directory of Open Access Journals (Sweden)

    Lalatendu Keshary Das

    2014-04-01

    Full Text Available Aim: To determine the metabolizable protein (MP content of common indigenous feedstuffs used in ruminant nutrition using in situ method. Materials and Methods: Nine ruminant feeds such as maize grain (MG, groundnut cake (GNC, mustard oilcake (MOC, cottonseed cake (CSC, deoiled rice bran (DORB, wheat bran (WB, berseem fodder (BF, maize fodder (MF and sorghum fodder (SF were included in this study. Each test feed was dried, ground and chemically analysed for proximate principles (DM, CP, EE, OM, Total ash, fiber fractions (NDF, ADF, cellulose, hemicellulose, lignin, NDICP and ADICP. Two adult fistulated bulls were used for evaluating the protein degradation characteristics of each test feed using the nylon bag method. Metabolizable energy (ME content of the test feeds were predicted from their chemical composition data using summative approach of NRC (2001 model. The equations of AFRC (1992 were used to predict the rumen degradable protein (RDP, digestible microbial protein (DMP, digestible undegraded feed protein (DUP and MP content of test feeds. Results: The MP content of MG, GNC, MOC, CSC, DORB, WB, BF, MF and SF was found to be 95.26, 156.41, 135.21, 125.06, 101.68, 107.11, 136.81, 72.01 and 76.65 g/kg DM, respectively. The corresponding ME (MJ/kg DM content of the test feeds was 13.66, 13.12, 13.65, 10.68, 9.08, 11.56, 9.64, 8.33 and 8.03, respectively. Among the test feeds, GNC contained the highest and MF contained the lowest MP per kg DM. Conclusion: It was concluded that the degradability of crude protein (CP of the test feeds can be used in MP determination and diet formulation. Feed CP content is not available as such at intestinal level in ruminants as a definite part of it undergoes extensive microbial degradation in rumen. The pattern and extent of such degradation do influence the amount of protein presented to lower digestive tract (MP for absorption and utilization in ruminants. It was also found that the MP content of a feed is

  2. Effects of alimet on nutrient digestibility, bacterial protein synthesis, and ruminal disappearance during continuous culture.

    Science.gov (United States)

    Vázquez-Añón, M; Cassidy, T; McCullough, P; Varga, G A

    2001-01-01

    A dual effluent continuous culture system was used to investigate the effects of inclusion of Alimet (Novus International, Inc., St. Louis, MO) feed supplement [an 88% aqueous solution of dl, 2-hydroxy-4-(methylthio) butanoic acid (HMB)] in the diet on nutrient digestibility, bacterial protein synthesis and ruminal disappearance of HMB. Four fermenters were fed three times daily a basal diet that consisted of 50% grain mixture and 50% forage for 9 d. In experiment 1, four concentrations of HMB (0, 0.20, 0.77, and 1.43% DM basis) were added to the diet and fed to the fermenters twice daily. In experiment 2, two concentrations of dietary HMB (0 and 0.88% DM basis) were fed twice daily and evaluated with two solids retention times (16.7 vs. 25.0 h) and two liquid dilution rates (0.15 vs. 0.125 h(-1)). Increasing the amount of HMB in the diet did not affect nutrient digestibility, volatile fatty acid concentrations, or ruminal escape of HMB. Bacterial protein synthesis was improved with the addition of HMB during high and low retention times. The extent of HMB escaping ruminal degradation ranged from 21.6 to 43.2% and was highest at the lower retention time. It can be concluded that a fraction of HMB survives rumen microbial degradation and, therefore, provides a rumen-protected form of methionine at the same time that it improves bacterial protein synthesis.

  3. Níveis de proteína degradável para novilhas em crescimento sobre a concentração de protozoários ciliados e outros parâmetros ruminais Levels of degradable protein given to heifers for the ciliate protozoa concentration and other ruminal parameters

    Directory of Open Access Journals (Sweden)

    Joaquim Balcells

    2001-05-01

    Full Text Available Em um ensaio de controle de parâmetros ruminais (pH, ritmo de trânsito (%h, volume ruminal (l, e concentração de protozoários ciliados, foram utilizadas quatro novilhas Holstein, canuladas no rúmen e duodeno, recebendo dietas com 4 níveis de proteína degradável, sem suplementar ou suplementadas com 25, 50 e 75 gramos de proteína degradável por quilo de concentrado em forma de uréia e caseína (98,2 e 93,2% de matéria orgânica e proteína bruta, respectivamente. As rações experimentais foram compostas por 25% de palha de cevada sem tratar e 75% de concentrado formulado em base a diferentes proporções (25/75 e 75/25 de milho/cevada, com 5% de resíduo de trigo e 5% de suplemento vitamínico mineral. Uma contínua infusão de marcador de fluxo de fase líquida (Cr-EDTA foi executada. Os oito tratamentos experimentais (2 dietas x 4 níveis de infusão foram administrados em 8 períodos de balanço em um delineamento cross-over, em esquema fatorial. O nível de infusão de N degradável afetou de forma significativa os valores médios ponderados de pH, que decresceram ao aumentar a suplementação (p 3/mL que de cevada (59,5 x 103/mL; p Entodinium alcançou 87,8% da população total. O gênero Epidinium apresentou menores valores com a dieta de cevada (3,73 vs. 1,89x103/mL, p 3/mL, p 5/mL, p In this experiment four Holstein heifers fitted with rumen and duodenum canulae were utilized to evaluate the rumen parameters as pH, passage rate (%/h, volume (L and ciliated protozoa concentration, when fed diets with four levels of rumen degradable protein. This was achieved supplementing 0, 25, 50 or 75 grams of degradable protein per kg of concentrate as a urea and casein (98.2% OM and 93.2% CP. The diets had 25% of barley straw without treatment and 75% concentrate formulated to contain different proportions (25/75 and 75/25 of corn and barley and also 5% of wheat residue and 5% of mineral and vitamin supplement. A continuous infusion of

  4. Protein value of cereals and cereal by-products for ruminants: a comparison between crude protein and protein-based estimates.

    Science.gov (United States)

    González, Javier; Mouhbi, Rabiaa; Guevara-González, Jesús Alberto; Rodríguez, Carlos Alberto; Arroyo, José María

    2015-01-01

    In situ estimates of ruminal undegraded fraction (RU) and effective intestinal digestibility (EID, corrected for microbial colonisation) of dry matter (DM), crude protein (CP) and total analysed amino acids (TAA) of rye, wheat and corn grains, wheat bran, wheat and barley distillers' dried grains with solubles (DDGS) and corn gluten feed were measured on three rumen and duodenum cannulated wethers using (15)N labelling techniques and considering ruminal rates of particle comminution (kc) and outflow. Results indicate that not considering kc and microbial colonisation led to considerable overestimations of RU which increased with feed ruminal degradation. Microbial colonisation may be also associated with overestimations of EID, whose estimates for DM, CP and TAA were predicted from parameters related with the ruminal escape of intestinally indigestible materials. The RU estimates were higher for TAA than for CP in grains, but the opposite was observed in by-products, whereas EID estimates were higher for TAA in all feeds. To obtain accurate protein values in these feedstuffs, it is required to consider both kc and ruminal microbial colonisation. The CP-based results underestimate the intestinally digested protein in grains and the opposite is evidenced in cereal by-products. Microbial protein synthesised in the rumen is largely the major fraction of the feedstuff protein value with the exception of DDGS.

  5. Combinações de fontes de nitrogênio e amido de alta e baixa degradabilidade ruminal avaliadas pela técnica in situ Combinations of starch and nitrogen sources in high and low ruminal degradability evaluated by in situ technique

    Directory of Open Access Journals (Sweden)

    Ivanor Nunes do Prado

    1998-10-01

    Full Text Available Este trabalho teve por objetivo avaliar a degradabilidade ruminal de quatro concentrados, os quais diferiam entre si pela diferença na degradação ruminal das fontes de amido e nitrogênio utilizadas. Foram utilizadas três vacas da raça Holandesa, com peso médio de 530kg e portadoras de cânula ruminal. Os concentrados formulados com fonte de amido e nitrogênio de alta degradabilidade ruminal apresentaram maiores valores de degradabilidade efetiva (DE da matéria seca, proteína bruta e amido para taxas de passagem de 2%/h, 5%/h e 8%/h e o inverso foi observado para os concentrados com fontes de amido e nitrogênio de baixa degradabilidade ruminal. Os concentrados formulados alternando as fontes de amido e de nitrogênio de alta e baixa degradabilidade ruminal apresentaram valores intermediários de DE da matéria seca e da proteína. A DE do amido foi influenciada pela fonte de amido independentemente da fonte de nitrogênio.The objective of this paper was to evaluate the ruminal degradability of four isonitrogenized and isoenergetic concentrates which differed as for starch and nitrogen sources. Three holstein cows with 530kg average weight and fistulated rumen were used in this investigation. The concentrates formulated with starch and nitrogen sources of high ruminal degradability presented higher values of drymatter, crude protein and starch effective degradability (ED for 2%/h, 5%/h and 8%/h passage rates, but the reverse was also observed for concentrates with starch and nitrogen sources of low ruminal degradability. The concentrates formulated by alternating starch and nitrogen sources of high and low ruminal degradability showed dry matter and protein ED intermediate values. The starch ED was influenced by the starch source independently of the nitrogen source.

  6. Oven-drying reduces ruminal starch degradation in maize kernels

    NARCIS (Netherlands)

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

    2014-01-01

    The degradation of starch largely determines the feeding value of maize (Zea mays L.) for dairy cows. Normally, maize kernels are dried and ground before chemical analysis and determining degradation characteristics, whereas cows eat and digest fresh material. Drying the moist maize kernels

  7. Effect of grass species on NDF ruminal degradability and ...

    African Journals Online (AJOL)

    uzivatel

    For the evaluation of rumen degradation parameters and INDF, grass species, year, ... variability in the chemical composition and degradability parameters, reflecting a wide range in NDF quality .... silage processing and microorganisms, may be more susceptible to enzymatic attack, mainly in ..... of their rapid maturing rate.

  8. Fractions of ruminant feeds: kinetics of degradation in vitro

    NARCIS (Netherlands)

    Azarfar, A.

    2007-01-01

    A widely adopted procedure to characterise the degradation in the rumen and its dynamics is the in situ incubation technique that assumes the washable fraction of feeds (W) to be equal to the soluble (S) fraction and that both are rapidly and completely degraded which may not be

  9. Proteins Profile in Milk from Three Species of Ruminants

    Directory of Open Access Journals (Sweden)

    Anamaria COZMA

    2011-03-01

    Full Text Available Milk proteins, caseins and whey proteins, are very important nutritionally, as they contain all essential aminoacids in optimal proportions and are the most important source of bioactive peptides. These peptides are protein fragments resulting from enzymatic hydrolysis of milk proteins, which carry numerous beneficial effects on the cardiovascular, nervous, gastrointestinal and immune systems. In this research, total proteins, caseins and whey proteins respectively, were dosed in milk from three species of ruminants-cow, goat and sheep, using a very sensitive method, Bradford photometric method. The highest content of total proteins was obtained in sheeps� milk (65.92 mg/ml and the lowest in cows� milk (40.03 mg/ml, intermediate values occurring in goats� milk (46.79 mg/ml. The lowest amount of caseins was found in cow milk (28.26 mg/ml, followed by sheep milk (42.55 mg/ml and goat milk (44.03 mg/ml. When the case of whey proteins, the highest values occur in sheep milk (23.36 mg/ml and the lowest in cow milk (11.79 mg/ml, goat milk having intermediate values (17.7 mg/ml. The results obtained indicate the dependence of protein concentration in milk of the studied ruminant species and stresses the importance of including goat and sheep milk in daily diet, along with cow milk, for an optimal intake of protein.

  10. Proteins Profile in Milk from Three Species of Ruminants

    Directory of Open Access Journals (Sweden)

    Anamaria COZMA

    2011-03-01

    Full Text Available Milk proteins, caseins and whey proteins, are very important nutritionally, as they contain all essential aminoacids in optimal proportions and are the most important source of bioactive peptides. These peptides are protein fragments resulting from enzymatic hydrolysis of milk proteins, which carry numerous beneficial effects on the cardiovascular, nervous, gastrointestinal and immune systems. In this research, total proteins, caseins and whey proteins respectively, were dosed in milk from three species of ruminants-cow, goat and sheep, using a very sensitive method, Bradford photometric method. The highest content of total proteins was obtained in sheeps milk (65.92 mg/ml and the lowest in cows milk (40.03 mg/ml, intermediate values occurring in goats milk (46.79 mg/ml. The lowest amount of caseins was found in cow milk (28.26 mg/ml, followed by sheep milk (42.55 mg/ml and goat milk (44.03 mg/ml. When the case of whey proteins, the highest values occur in sheep milk (23.36 mg/ml and the lowest in cow milk (11.79 mg/ml, goat milk having intermediate values (17.7 mg/ml. The results obtained indicate the dependence of protein concentration in milk of the studied ruminant species and stresses the importance of including goat and sheep milk in daily diet, along with cow milk, for an optimal intake of protein.

  11. Acute phase protein response during acute ruminal acidosis in cattle

    DEFF Research Database (Denmark)

    Danscher, A. M.; Thoefner, M. B.; Heegaard, Peter M. H.

    2011-01-01

    acids. In humans, inflammation has been linked to metabolic diseases. In cattle, studies into the possible links between acid-base changes, inflammation/innate immunity and metabolic disease are warranted as this might improve our understanding of the production disease complexes occurring in particular......The aim of the study was to describe the acute phase protein and leukocyte responses in dairy heifers during acute, oligofructose-induced ruminal acidosis. The study included 2 trials involving oral oligofructose overload (17g/kg BW) to nonpregnant Danish Holstein heifers. Trial 1 included 12...... than control heifers at 18 and 24h after overload (max. 13.7±4.3 billions/L). Feeding had no effect on plasma fibrinogen concentrations or WBC in Trial 1.Acute ruminal and systemic acidosis caused by oligofructose overload resulted in distinct acute phase protein and leukocyte responses in dairy...

  12. The Roles of Syncytin-Like Proteins in Ruminant Placentation

    Directory of Open Access Journals (Sweden)

    Yuki Nakaya

    2015-06-01

    Full Text Available Recent developments in genome sequencing techniques have led to the identification of huge numbers of endogenous retroviruses (ERV in various mammals. ERVs, which occupy 8%–13% of mammalian genomes, are believed to affect mammalian evolution and biological diversity. Although the functional significance of most ERVs remains to be elucidated, several ERVs are thought to have pivotal roles in host physiology. We and other groups recently identified ERV envelope proteins (e.g., Fematrin-1, Syncytin-Rum1, endogenous Jaagsiekte sheep retrovirus Env that may determine the morphogenesis of the unique fused trophoblast cells, termed trinucleate cells and syncytial plaques, found in ruminant placentas; however, there are still a number of outstanding issues with regard to the role of ERVs that remain to be resolved. Here, we review what is known about how these ERVs have contributed to the development of ruminant-specific trophoblast cells.

  13. 21 CFR 589.2000 - Animal proteins prohibited in ruminant feed.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Animal proteins prohibited in ruminant feed. 589... Animal proteins prohibited in ruminant feed. (a) Definitions—(1) Protein derived from mammalian tissues means any protein-containing portion of mammalian animals, excluding: Blood and blood products; gelatin...

  14. Effect of grass species on NDF ruminal degradability and ...

    African Journals Online (AJOL)

    uzivatel

    ryegrass (Lolium perenne L.; 10%), white clover (Trifolium repens L.; 10%) and herbs (5%). For the in situ determination, the silage samples were oven-dried at 50 °C for 48 h and milled to pass a 1 mm screen. The rumen degradability of the NDF, in the experimental silages, was determined by incubating 1.5 g of each.

  15. Ruminal dry matter degradability of treated soybean meal as source ...

    African Journals Online (AJOL)

    In order to study rumen degradation of dry matter for treating soybean meal with black liquor as source of xylose and microwave radiation, an experiment in nylon bag technique was performed. Samples for treating soybean meal for 0, 2, 4, 8, 16, 24 and 48 h in the rumen of three Taleshi male cows were incubated. Soybean ...

  16. Substitution of common concentrates with by-products modulated ruminal fermentation, nutrient degradation, and microbial community composition in vitro

    National Research Council Canada - National Science Library

    Ertl, P; Knaus, W; Metzler-Zebeli, B U; Klevenhusen, F; Khiaosa-Ard, R; Zebeli, Q

    2015-01-01

    ...) with a mixture consisting solely of by-products from the food industry (BP) at 2 different forage-to-concentrate ratios on ruminal fermentation profile, nutrient degradation, and abundance of rumen microbiota...

  17. Digestibilidade e balanço de nitrogênio de rações com diferentes teores de proteína degradável no rúmen e milho moído como fonte de amido em ovinos Digestibility and nitrogen balance of sheep diets containing different levels of ruminal degradable protein and ground corn

    Directory of Open Access Journals (Sweden)

    Lúcia Maria Zeoula

    2006-10-01

    Full Text Available O objetivo neste trabalho foi avaliar a digestibilidade aparente de rações com diferentes teores de proteína degradável no rúmen (PDR e fonte de amido de baixa degradabilidade ruminal (milho moído em ovinos. Utilizaram-se quatro ovinos castrados, sem raça definida (SRD, com peso vivo médio de 38,5 kg, em um delineamento experimental quadrado latino 4 x 4. As quatro rações experimentais utilizadas foram compostas de 75% de volumoso (feno de aveia e 25% de concentrado à base de milho moído (MM, farelo de soja (FS, farelo de algodão (FA, farinha de peixe (FP, uréia e sal mineral, compostas com quatro teores de PDR (46, 50, 54 e 58% de PDR nas rações. Determinaram-se a digestibilidade aparente total dos nutrientes e o balanço de nitrogênio nos animais alimentados com as rações experimentais. Os consumos de MS, MO, PB, amido, FDN e FDA não diferiram entre as rações testadas. Da mesma forma, o nitrogênio ingerido (NI, o N excretado nas fezes e na urina, em g/dia, o balanço de nitrogênio (%NI e os coeficientes de digestibilidade da MS, MO, PB, FDN e FDA não diferiram entre as rações experimentais, sendo estimados valores médios de 31,68; 6,34; 11,31 g/dia e 44,27% NI, respectivamente, e de 71,90; 73,00; 79,70; 61,10; e 52,00%, respectivamente. Independentemente dos teores de PDR em rações contendo milho como fonte de amido de baixa degradabilidade ruminal, não houve efeitos sobre a digestibilidade aparente dos nutrientes e o balanço de nitrogênio em ovinos.Four castrated sheep averaging 38.5 kg of body weight were randomly assigned to a 4 x 4 Latin square to evaluate the apparent digestibility of nutrients and nitrogen balance (NB on diets containing different levels of rumen degradable protein (RDP: 46, 50, 54, and 58% and ground corn. The four experimental diets were composed by 75% of forage (oat hay and 25% of concentrate containing ground corn (GC, soybean meal (SM, cottonseed meal (CM, fish meal (FM, urea, and

  18. Effects of ruminally degradable starch levels on performance, nitrogen balance, and nutrient digestibility in dairy cows fed low corn-based starch diets.

    Science.gov (United States)

    Luo, Guobin; Xu, Wenbin; Yang, Jinshan; Li, Yang; Zhang, Liyang; Wang, Yizhen; Lin, Cong; Zhang, Yonggen

    2017-05-01

    This trial was performed to examine the effects of ruminally degradable starch (RDS) levels in total mixed ration (TMR) with low corn-based starch on the milk production, whole-tract nutrient digestibility and nitrogen balance in dairy cows. Eight multiparous Holstein cows (body weight [BW]: 717±63 kg; days in milk [DIM]: 169±29) were assigned to a crossover design with two dietary treatments: a diet containing 62.3% ruminally degradable starch (% of total starch, low RDS) or 72.1% ruminally degradable starch (% of total starch, high RDS). Changes to the ruminally degradable levels were conducted by using either finely ground corn or steam-flaked corn as the starch component. The results showed that dry matter intake, milk yield and composition in dairy cows were not affected by dietary treatments. The concentration of milk urea nitrogen was lower for cows fed high RDS TMR than low RDS TMR. The whole-tract apparent digestibility of neutral detergent fiber, acid detergent fiber and crude protein decreased, and that of starch increased for cows fed high RDS TMR over those fed low RDS TMR, with no dietary effect on the whole-tract apparent digestibility of dry matter and organic matter. The proportion of urinary N excretion in N intake was lower and that of fecal N excretion in N intake was higher for cows fed high RDS TMR than those fed low RDS TMR. The N secretion in milk and the retention of N were not influenced by the dietary treatments. Total purine derivative was similar in cows fed high RDS TMR and low RDS TMR. Consequently, estimated microbial N flow to the duodenum was similar in cows fed high RDS TMR and low RDS TMR. Results of this study show that ruminally degradable starch levels can influence whole-tract nutrient digestibility and nitrogen balance in dairy cows fed low corn-based starch diets, with no influence on performance.

  19. In situ degradation of almond (Prunus dulcis L.) hulls, a potential feed material for ruminants

    OpenAIRE

    JAFARI, SAEID; Alizadeh, Alireza; IMNI, ALI; Meng, Goh Yong; Rajion, Mohamed Ali; EBRAHIMI, Mahdi

    2015-01-01

    The nutritive values of almond hulls (Ahs), alfalfa (ALF), and sugar beet pulp (SBP) were determined using the in situ nylon bag technique. The ruminal dry matter (DM) degradation kinetics of 4 varieties of Ahs were compared with ALF and SBP. The almond varieties tested were Rabbi (RAB), Mamaii (MAM), Shahrud 15 (SH15), and Shokufe (SH). Samples were incubated in triplicate at 0, 4, 8, 12, 16, 24, 48, 72, and 96 h in 3 rumen-fistulated steers. The average total degradability fraction (TDF) of...

  20. Degradabilidade ruminal da matéria seca e proteína bruta, e tempo de colonização microbiana de oleaginosas, utilizadas na alimentação de ovinos - doi: 10.4025/actascianimsci.v33i4.11388 Ruminal degradability of dry matter and crude protein, and microbial colonization time of oil grains in sheep feeding - doi: 10.4025/actascianimsci.v33i4.11388

    Directory of Open Access Journals (Sweden)

    Euclides Reuter de Oliveira

    2011-09-01

    Full Text Available Avaliou-se a degradabilidade in situ, dos grãos de linhaça, canola, colza e milho, caroço de algodão, e dos concentrados casca e farelo de soja, em ovinos. Foram utilizados três animais da raça Santa Inês, fistulados e providos de cânulas ruminais, mantidos em baias individuais, recebendo diariamente capim picado e ração concentrada. Os alimentos foram incubados em ordem decrescente de 72, 48, 24, 18, 6, 3 e 0h. O teor de proteína dos alimentos avaliados foram 34,77; 30,07; 23,70; 10,64; 26,12; 14,65 e 56,90% para canola, colza, linhaça, milho, caroço de algodão, casca de soja e farelo de soja, respectivamente. A canola e a colza apresentaram baixa degradabilidade efetiva para a MS, com valor médio de 33,68%. O grão de linhaça apresentou degradabilidade efetiva para a MS de 64,24%, com fração potencialmente degradável de 87,89%. O milho apresentou fração solúvel de 12,33% e uma degradação de 39,67% para a MS. O farelo de soja apresentou-se dentro dos parâmetros normais de degradação com 52,61% para a MS e 52,83% para PB. Para a linhaça, canola e colza a degradabilidade da proteína apresentou valor médio de 18,34%. Os grãos de avaliados apresentaram baixa degradabilidade efetiva para a matéria seca e proteína bruta.This study evaluated in situ ruminal degradability of grains of linseed, canola, rapeseed and corn, whole cottonseed, as well as soybean hulls and soybean meal, in sheep. Three Santa Inês sheep were fistulated and fitted with rumen cannulas. The animals were housed in individual stalls, receiving chopped grass and concentrated feed daily. Feeds were incubated in descending order of 72, 48, 24, 18, 6, 3 and 0h. Protein content was 34.77% for canola, 30.07% for rapeseed, 23.70% for linseed, 10.64% for corn, 26.12% for cottonseed, 14.65% for soybean hulls, and 56.90% for soybean meal. Canola and rapeseed showed low effective degradability of DM, with mean value of 33.68%. Linseed grain had DM

  1. Ruminal degradability of alfalfa in the Costa Rica’s Central Valley, Eastern zone

    Directory of Open Access Journals (Sweden)

    Carlos Boschini Figueroa

    2017-09-01

    Full Text Available The objective of this work was to evaluate in dairy cows, the potential of ruminal degradability of four alfalfa (Medicago sativa varieties developed in Argentina. The research was carried out in the Alfredo Volio Mata Experimental Station, located in the province of Cartago, Costa Rica; the area has a mean temperature of 17.9°C; a height of 1542 masl, and 1465.9 mm of rain evenly distributed between the months of May and November. Sampling was performed during 2014’s rainy season with ground level harvest of alfalfa, with samples of the varieties of Mora, Patricia, Victoria and Super Monarca, with harvest age of 28, 35, 42, 49, 56 and 63 days; all samples were dried, minced and incubated by duplicate on two Jersey-Reyna ruminally fistulated cows, for 0, 3, 6, 12, 24 and 48 hours. The results were analyzed through the Marquardt algorithm with the SAS software. Statical differences were only found between soluble fraction and degradation rate (p<0.05 according to harvest ages, with values of soluble, potentially degradable and degradation rate of 35.34%-44.65%, 36.46%-39.96% and 0.0734/h-0.1176/h, respectively. Likewise, effective degradability percentages of 59.06%-67.09%, with passage rates of 6%/h, according to harvest ages were obtained. Given the obtained results, the analyzed varieties of M. sativa showed a digestible forage, which could be used for the feeding of highly production animals.

  2. Effect of roasting on ruminal degradation, intestinal digestibility and ...

    African Journals Online (AJOL)

    Even though CSOC could be roasted longer at the same temperature than SBOC (90 vs.45 min at 130'C) to achieve similar ADIN values, the effective protein degrada- tion rate remained higher in the case of CSOC. The CP disappearance from polyester bags and the UDP-D are shown in Table 4. The mean retention times ...

  3. Influence of exogenous fibrolytic enzymes on in vitro and in sacco degradation of forages for ruminants

    Directory of Open Access Journals (Sweden)

    Lorenzo Carreón

    2010-02-01

    Full Text Available An in vitro assay was carried out to evaluate the effects of exogenous fibrolytic enzymes (1, 2, 3 and 4 g/kg DM powder preparation containing xylanase and cellulase from Aspergillus niger and Trichoderma viride on DM, NDF and ADF degradation of alfalfa hay, corn silage, corn stover, elephant grass, Guinea grass and oat straw. Kinetics data of in vitro degradations were analyzed. The potentially degradable fraction and degradation rate of NDF and ADF of alfalfa increased quadratically (P<0.05 as the inclusion level of enzyme increased up to 3 g. The others forages were not affected by the enzyme. An in sacco trail was performed using four Holstein steers fitted with ruminal cannulas to evaluate the effects of the exogenous fibrolytic enzymes (3 g/kg DM on DM, NDF and ADF degradation of alfalfa hay and corn stover. Kinetics data were also analyzed. The potentially degradable fraction degradation of NDF (62.0 vs 65.7% and ADF (52.8 vs 56.9%, of alfalfa hay were increased (P<0.05 by the exogenous fibrolytic enzymes, but no differences were found for corn stover. These results suggest that the enzymes increased in vitro and in sacco fibre degradation only for alfalfa hay.

  4. Multiple cellobiohydrolases and cellobiose phosphorylases cooperate in the ruminal bacterium Ruminococcus albus 8 to degrade cellooligosaccharides

    Science.gov (United States)

    Devendran, Saravanan; Abdel-Hamid, Ahmed M.; Evans, Anton F.; Iakiviak, Michael; Kwon, In Hyuk; Mackie, Roderick I.; Cann, Isaac

    2016-10-01

    Digestion of plant cell wall polysaccharides is important in energy capture in the gastrointestinal tract of many herbivorous and omnivorous mammals, including humans and ruminants. The members of the genus Ruminococcus are found in both the ruminant and human gastrointestinal tract, where they show versatility in degrading both hemicellulose and cellulose. The available genome sequence of Ruminococcus albus 8, a common inhabitant of the cow rumen, alludes to a bacterium well-endowed with genes that target degradation of various plant cell wall components. The mechanisms by which R. albus 8 employs to degrade these recalcitrant materials are, however, not clearly understood. In this report, we demonstrate that R. albus 8 elaborates multiple cellobiohydrolases with multi-modular architectures that overall enhance the catalytic activity and versatility of the enzymes. Furthermore, our analyses show that two cellobiose phosphorylases encoded by R. albus 8 can function synergistically with a cognate cellobiohydrolase and endoglucanase to completely release, from a cellulosic substrate, glucose which can then be fermented by the bacterium for production of energy and cellular building blocks. We further use transcriptomic analysis to confirm the over-expression of the biochemically characterized enzymes during growth of the bacterium on cellulosic substrates compared to cellobiose.

  5. Ruminal degradability and summative models evaluation for total digestible nutrients prediction of some forages and byproducts in goats.

    Science.gov (United States)

    Rosendo, Oswaldo; Freitez, Luis; López, Rafael

    2013-01-01

    In in vitro true dry matter degradability (IVTDMD), in situ dry matter degradability, and neutral detergent fiber degradability, both in vitro (IVNDFD) and in situ (ISNDFD) techniques were used with crossbred goats to determine dry matter and neutral detergent fiber (NDF) ruminal degradability in eight forages and four industrial byproducts. Total digestible nutrients (TDN) content obtained with five different summative models (summative equations) were studied to compare the precision of estimates. All these models included digestible fractions of crude protein, ether extract, and nonfiber carbohydrates that were calculated from chemical composition, but digestible NDF (dNDF) was obtained from IVNDFD (IVdNDF), ISNDFD (ISdNDF), or by using the Surface Law approach. On the basis of the coefficient of determination (R (2)) of the simple lineal regression of predicted TDN (y-axes) and observed IVTDMD (x-axes), the precision of models was tested. The predicted TDN by the National Research Council model exclusively based on chemical composition only explains up to 41% of observed IVTDMD values, whereas the model based on IVdNDF had a high precision (96%) to predict TDN from forage and byproducts fiber when used in goats.

  6. Effects of type of canola protein supplement on ruminal fermentation and nutrient flow to the duodenum in beef heifers.

    Science.gov (United States)

    Gozho, G N; McKinnon, J J; Christensen, D A; Racz, V; Mutsvangwa, T

    2009-10-01

    Ruminal fermentation, nutrient digestion, and flows to the duodenum in growing cattle fed differently produced canola protein supplements were studied in a 4 x 4 Latin square design using Speckle Park heifers (initial BW = 451 +/- 26 kg). Canola protein supplement treatments consisted of the following: 1) 8.78% regular canola meal (RCM); 2) 9.25% RCM plus 1.80% canola oil (RCMO); 3) 11.1% canola presscake from biodiesel oil extraction (CPC); and 4) 8.14% high ruminally undegradable protein (RUP) canola meal (RUCM) plus 1.32% canola oil (RUCMO). Experimental diets also contained 39.9, 40.2, 39.9, and 39.9% barley grain; 31.7, 31.4, 31.2, and 31.4% barley silage; and 17.5, 15.2, 15.6, and 16.5% oat hulls for the RCM, RCMO, CPC, and RUCMO diets, respectively. Feeding the CPC, RCMO, and RUCMO diets decreased (P canola oil in the RCMO diet or residual oil in the CPC diet resulted in greater ruminal concentrations of propionate (P canola oil to the RCMO and RUCMO dietary treatments decreased the digestibility of ADF (P < or = 0.08) and NDF (P < or = 0.08) in the total tract compared with the RCM diet. Total tract digestibility of OM was also decreased (P = 0.02) in heifers fed the RUCMO compared with the RCM diet. Notwithstanding the different processing methods employed in making RCM, CPC, or RUCM, there were no differences among the diets for ruminally degraded protein, ruminal microbial protein synthesis, and the flow of N fractions to the duodenum.

  7. Accounting for the effects of a ruminal nitrogen deficiency within the structure of the Cornell Net Carbohydrate and Protein System.

    Science.gov (United States)

    Tedeschi, L O; Fox, D G; Russell, J B

    2000-06-01

    The Cornell Net Carbohydrate and Protein System (CNCPS) prediction of fiber digestion and microbial mass production from ruminally degraded carbohydrate has been adjusted to accommodate a ruminal N deficiency. The steps for the adjustment are as follows: 1) the ruminal available peptide and ammonia pools are used to determine the N allowable microbial growth; 2) this value is subtracted from the energy allowable microbial growth to obtain the reduction in microbial mass; 3) this mass reduction is allocated between pools of bacteria digesting fiber (FC) and nonfiber (NFC) carbohydrate according to their original proportions in the energy allowable microbial growth; 4) the reduction in fermented FC is computed as the FC bacterial mass reduction divided by its yield (g bacteria/g FC digested); and 5) this reduction is added to the FC fraction escaping the rumen. Five published studies included information that allowed us to evaluate the response of animals to added dietary N. These evaluations compared observed and CNCPS-predicted ADG with and without this adjustment. The adjustment decreased the CNCPS overprediction of ADG from 19.2 to 4.7%, mean bias declined from .16 to .04 kg/d, and the r2 of the regression between observed and metabolizable energy (ME) or metabolizable protein allowable ADG was increased from .83 to .88 with the adjustment. When the observed dry matter intake was regressed against CNCPS-predicted DMI with an adjustment for reduction in cell wall digestibility, the r2 was increased from .77 to .88. These results indicated the adjustment for ruminal nitrogen deficiency increased the accuracy of the CNCPS model in evaluating diets of growing animals when ruminally degraded N is deficient.

  8. Effect of protein supplementation on ruminal parameters and microbial community fingerprint of Nellore steers fed tropical forages.

    Science.gov (United States)

    Bento, C B P; Azevedo, A C; Gomes, D I; Batista, E D; Rufino, L M A; Detmann, E; Mantovani, H C

    2016-01-01

    In tropical regions, protein supplementation is a common practice in dairy and beef farming. However, the effect of highly degradable protein in ruminal fermentation and microbial community composition has not yet been investigated in a systematic manner. In this work, we aimed to investigate the impact of casein supplementation on volatile fatty acids (VFA) production, specific activity of deamination (SAD), ammonia concentration and bacterial and archaeal community composition. The experimental design was a 4×4 Latin square balanced for residual effects, with four animals (average initial weight of 280±10 kg) and four experimental periods, each with duration of 29 days. The diet comprised Tifton 85 (Cynodon sp.) hay with an average CP content of 9.8%, on a dry matter basis. Animals received basal forage (control) or infusions of pure casein (230 g) administered direct into the rumen, abomasum or divided (50 : 50 ratio) in the rumen/abomasum. There was no differences (P>0.05) in ruminal pH and microbial protein concentration between supplemented v. non-supplemented animals. However, in steers receiving ruminal infusion of casein the SAD and ruminal ammonia concentration increased 33% and 76%, respectively, compared with the control. The total concentration of VFA increased (P0.05) in species richness and diversity of γ-proteobacteria, firmicutes and archaea between non-supplemented Nellore steers and steers receiving casein supplementation in the rumen. However, species richness and the Shannon-Wiener index were lower (P<0.05) for the phylum bacteroidetes in steers supplemented with casein in the rumen compared with non-supplemented animals. Venn diagrams indicated that the number of unique bands varied considerably among individual animals and was usually higher in number for non-supplemented steers compared with supplemented animals. These results add new knowledge about the effects of ruminal and postruminal protein supplementation on metabolic activities of

  9. Evaluation of ruminal degradation profiles of forages using bags made from different textiles

    Directory of Open Access Journals (Sweden)

    Tiago Neves Pereira Valente

    2011-11-01

    Full Text Available The objective of this study was to evaluate the in situ degradation profiles of dry matter (DM and neutral detergent fiber (NDF of different forages using nylon (50 µm, F57 (Ankom® and non-woven textile (NWT - 100 g/m² bags. Eight forage samples were used: sugarcane, corn silage, elephant grass cut at 50 and 250 days of regrowth, corn straw, signal grass hay, coast cross hay, and fresh alfalfa. Samples were incubated for 0, 3, 6, 12, 18, 24, 48, 72, 96, 120, 144, 168, 192, 216, 240, and 312 hours. Two bags of each textile were used at each incubation time, totaling 768 bags, using two crossbred Holstein × Zebu steers fitted with ruminal canullae. There was difference in the common rate of lag and degradation (λ of DM for all forages, except for sugarcane. In general, higher λ estimates were obtained using nylon, followed by NWT and F57. Concerning NDF degradation profiles, differences in λ were observed for all forages. Greater estimates were obtained using nylon. Degradation profiles of DM and NDF must not be evaluated using F57 and NWT. These textiles underestimate the degradation rate due to constraints regarding exchange between bags' content and rumen environment.

  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. The Prospect of Hydrolyzed Feather Meal as Ruminant Feeds Through Protein Quality Improvement by Microbes

    Directory of Open Access Journals (Sweden)

    CH Prayitno

    2003-01-01

    Full Text Available The waste of the broiler processing (feather is a potential source for animal feed. However the presence of keratins cause limited of feather use. Before using, therefore, feather must be treated to hydrolyze cysteine disulfide bound dominating keratins protein. Enzymatic (biological treatment using microbes will produce specific feather hydrolyzed and does not have negative impact on environment. The research objected to get the microbes which degradated selected keratins, improve protein quality of feather meal and find out the best ration formulation true in vitro the basic information to formulate in vivo ration. The research has been done in Laboratory of Animal Feedstuff Faculty of Animal Science UNSOED for eight months. Fermentation trial was done on liquid media with bath system. In vitro trial used of Tilley and Terry methods with parameter observe was dry matter digestibility, organic matter digestibility, protein degradation, total VFA and solubility in pepsin. Based on all parameter, on fermentation trial with Bacillus licheniformis decides broiler chicken feather had good prospect to be developed on feed protein source. In vitro trial recommended ration with formulation of fermented feather meal concentrate (15 percent, soybeans meal (5 percent, rice bran (20 percent, molasses (4 percent, mineral mix (1 percent, with forage: concentrate ratio 40 : 60 could be used as in vivo ration. (Animal Production 5(1: 19-24 (2003   Key words : Hydrolyze, Feather, Keratin, Digestibility, Ruminant

  12. Unique aspects of fiber degradation by the ruminal ethanologen Ruminococcus albus 7 revealed by physiological and transcriptomic analysis.

    Science.gov (United States)

    Christopherson, Melissa R; Dawson, John A; Stevenson, David M; Cunningham, Andrew C; Bramhacharya, Shanti; Weimer, Paul J; Kendziorski, Christina; Suen, Garret

    2014-12-04

    Bacteria in the genus Ruminococcus are ubiquitous members of the mammalian gastrointestinal tract. In particular, they are important in ruminants where they digest a wide range of plant cell wall polysaccharides. For example, Ruminococcus albus 7 is a primary cellulose degrader that produces acetate usable by its bovine host. Moreover, it is one of the few organisms that ferments cellulose to form ethanol at mesophilic temperatures in vitro. The mechanism of cellulose degradation by R. albus 7 is not well-defined and is thought to involve pilin-like proteins, unique carbohydrate-binding domains, a glycocalyx, and cellulosomes. Here, we used a combination of comparative genomics, fermentation analyses, and transcriptomics to further clarify the cellulolytic and fermentative potential of R. albus 7. A comparison of the R. albus 7 genome sequence against the genome sequences of related bacteria that either encode or do not encode cellulosomes revealed that R. albus 7 does not encode for most canonical cellulosomal components. Fermentation analysis of R. albus 7 revealed the ability to produce ethanol and acetate on a wide range of fibrous substrates in vitro. Global transcriptomic analysis of R. albus 7 grown at identical dilution rates on cellulose and cellobiose in a chemostat showed that this bacterium, when growing on cellulose, utilizes a carbohydrate-degrading strategy that involves increased transcription of the rare carbohydrate-binding module (CBM) family 37 domain and the tryptophan biosynthetic operon. Our data suggest that R. albus 7 does not use canonical cellulosomal components to degrade cellulose, but rather up-regulates the expression of CBM37-containing enzymes and tryptophan biosynthesis. This study contributes to a revised model of carbohydrate degradation by this key member of the rumen ecosystem.

  13. Ruminal tryptophan-utilizing bacteria degrade ergovaline from tall fescue seed extract.

    Science.gov (United States)

    Harlow, B E; Goodman, J P; Lynn, B C; Flythe, M D; Ji, H; Aiken, G E

    2017-02-01

    The objectives of this study were to evaluate degradation of ergovaline in a tall fescue [ (Schreb.) Darbysh.] seed extract by rumen microbiota ex vivo and to identify specific bacteria capable of ergovaline degradation in vitro. Rumen cell suspensions were prepared by harvesting rumen fluid from fistulated wether goats ( = 3), straining, and differential centrifugation. Suspensions were dispensed into anaerobic tubes with added Trypticase with or without extract (∼10 μg kg ergovaline). Suspensions were incubated for 48 h at 39°C. Samples were collected at 0, 24, and 48 h for ergovaline analysis and enumeration of hyper-ammonia producing (HAB) and tryptophan-utilizing bacteria. Ergovaline values were analyzed by repeated measures using the mixed procedure of SAS. Enumeration data were log transformed for statistical analysis. When suspensions were incubated with extract, 11 to 15% of ergovaline disappearance was observed over 48 h ( = 0.02). After 24 h, suspensions with added extract had 10-fold less HAB than controls ( = 0.04), but treatments were similar by 48 h ( = 1.00). However, after 24 h and 48 h, suspensions with extract had 10-fold more tryptophan-utilizing bacteria ( bacteria tested did not degrade ergovaline. The results of this study indicate which rumen bacteria may play an important role in ergovaline degradation and that microbiological strategies for controlling their activity could have ramifications for fescue toxicosis and other forms of ergotism in ruminants.

  14. Excretion pathways and ruminal disappearance of glyphosate and its degradation product aminomethylphosphonic acid in dairy cows.

    Science.gov (United States)

    von Soosten, D; Meyer, U; Hüther, L; Dänicke, S; Lahrssen-Wiederholt, M; Schafft, H; Spolders, M; Breves, G

    2016-07-01

    From 6 balance experiments with total collection of feces and urine, samples were obtained to investigate the excretion pathways of glyphosate (GLY) in lactating dairy cows. Each experiment lasted for 26d. The first 21d served for adaptation to the diet, and during the remaining 5d collection of total feces and urine was conducted. Dry matter intake and milk yield were recorded daily and milk and feed samples were taken during the sampling periods. In 2 of the 6 experiments, at the sampling period for feces and urine, duodenal contents were collected for 5d. Cows were equipped with cannulas at the dorsal sac of the rumen and the proximal duodenum. Duodenal contents were collected every 2h over 5 consecutive days. The daily duodenal dry matter flow was measured by using chromium oxide as a volume marker. All samples (feed, feces, urine, milk and duodenal contents were analyzed for GLY and aminomethylphosphonic acid (AMPA). Overall, across the 6 experiments (n=32) the range of GLY intake was 0.08 to 6.67mg/d. The main proportion (61±11%; ±SD) of consumed GLY was excreted with feces; whereas excretion by urine was 8±3% of GLY intake. Elimination via milk was negligible. The GLY concentrations above the limit of quantification were not detected in any of the milk samples. A potential ruminal degradation of GLY to AMPA was derived from daily duodenal GLY flow. The apparent ruminal disappearance of GLY intake was 36 and 6%. In conclusion, the results of the present study indicate that the gastrointestinal absorption of GLY is of minor importance and fecal excretion represents the major excretion pathway. A degradation of GLY to AMPA by rumen microbes or a possible retention in the body has to be taken into account. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  15. 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)

  16. Fontes protéicas e energéticas com diferentes degradabilidades ruminais para vacas em lactação - DOI: 10.4025/actascianimsci.v29i2.238 Protein (raw or rosted soybeans and energy (coursely ground or steamflaked corn sources with varying ruminal degradabilities for lactating cows - DOI: 10.4025/actascianimsci.v29i2.238

    Directory of Open Access Journals (Sweden)

    Hugo Imaizumi

    2007-11-01

    Full Text Available O objetivo deste trabalho foi avaliar a combinação de fontes de energia e proteína com diferentes degradabilidades ruminais. Foram utilizadas quatro vacas multíparas (23 kg leite dia-1 em Quadrado Latino 4 x 4 com arranjo fatorial 2 x 2: duas fontes de proteína (soja crua ou tostada e duas de energia (milho moído, grosso ou floculado. O tratamento com milho moído grosso e soja tostada resultou em maior consumo de matéria seca (p 0,05 o desempenho lactacional. A tostagem da soja e a floculação do milho diminuíram (p The objective of this trial was the evaluation of ruminal degradation synchrony of protein and energy sources. Four Holstein cows (23 kg day-1 were used in a 4 x 4 Latin Square design and 2 x 2 factorial arrangement: two protein (raw vs. roasted soybeans with two energy sources (coarsely ground vs. steam-flaked corn. Dry matter intake was higher for cows fed coarsely ground corn with roasted soybean (p 0.05. Rumen pH was not affected by treatments (p > 0.05. Both roasted soybeans and flaking of corn decreased rumen ammonia nitrogen (p < 0.01. Animal fed steamflaked corn tended showed higher total VFA molar concentrations (p = 0.10, but there were no treatments effect on ruminal pH. A higher (p = 0.06 ruminal starch digestibility was observed for raw soybean treatment.

  17. The Occurrence, Biosynthesis, and Molecular Structure of Proanthocyanidins and Their Effects on Legume Forage Protein Precipitation, Digestion and Absorption in the Ruminant Digestive Tract

    Science.gov (United States)

    Jonker, Arjan; Yu, Peiqiang

    2017-01-01

    Forages grown in temperate regions, such as alfalfa (Medicago sativa L.) and white clover (Trefolium repens L.), typically have a high nutritional value when fed to ruminants. Their high protein content and degradation rate result, however, in poor utilization of protein from the forage resulting in excessive excretion of nitrogen into the environment by the animal. Proanthocyanindins (also known as condensed tannins) found in some forage legumes such as birdsfoot trefoil (Lotus corniculatus L.), bind to dietary protein and can improve protein utilization in the animal. This review will focus on (1) the occurrence of proanthocyanidins; (2) biosynthesis and structure of proanthocyanidins; (3) effects of proanthocyanidins on protein metabolism; (4) protein precipitating capacity of proanthocyanidins and their effects on true intestinal protein adsorption by ruminants; and (5) effect on animal health, animal performance and environmental emissions. PMID:28531145

  18. The Occurrence, Biosynthesis, and Molecular Structure of Proanthocyanidins and Their Effects on Legume Forage Protein Precipitation, Digestion and Absorption in the Ruminant Digestive Tract

    Directory of Open Access Journals (Sweden)

    Arjan Jonker

    2017-05-01

    Full Text Available Forages grown in temperate regions, such as alfalfa (Medicago sativa L. and white clover (Trefolium repens L., typically have a high nutritional value when fed to ruminants. Their high protein content and degradation rate result, however, in poor utilization of protein from the forage resulting in excessive excretion of nitrogen into the environment by the animal. Proanthocyanindins (also known as condensed tannins found in some forage legumes such as birdsfoot trefoil (Lotus corniculatus L., bind to dietary protein and can improve protein utilization in the animal. This review will focus on (1 the occurrence of proanthocyanidins; (2 biosynthesis and structure of proanthocyanidins; (3 effects of proanthocyanidins on protein metabolism; (4 protein precipitating capacity of proanthocyanidins and their effects on true intestinal protein adsorption by ruminants; and (5 effect on animal health, animal performance and environmental emissions.

  19. Protein Repair and Degradation during Aging

    Directory of Open Access Journals (Sweden)

    Bertrand Friguet

    2002-01-01

    Full Text Available Cellular aging is characterized by a build-up of oxidatively modified proteins. The steady-state level of oxidized proteins depends on the balance between the rate of protein oxidative damage and the rates of protein degradation and repair. Therefore, the accumulation of oxidized protein with age can be due to increased protein damage, decreased oxidized protein degradation and repair, or the combination of both mechanisms. The proteasomal system is the major intracellular proteolytic pathway implicated in the degradation of oxidized protein, and the peptide methionine sulfoxide reductase catalyzes the reduction of methionine sulfoxide (i.e., oxidized methionine to methionine within proteins. A short summary on protein oxidative damage and oxidized protein degradation is given, and evidence for a decline of proteasome function with age is presented. Arguments for the implication of peptide methionine sulfoxide reductase in the age-related accumulation of oxidized protein are also discussed.

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

    Directory of Open Access Journals (Sweden)

    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.

  1. Recent advances in estimating protein and energy requirements of ruminants

    Science.gov (United States)

    Considerable efforts have been made in gathering scientific data and developing feeding systems for ruminant animals in the last 50 years. Future endeavours should target the assessment, interpretation, and integration of the accumulated knowledge to develop nutrition models in a holistic and pragma...

  2. Effects of varieties and timing of subsequent cutting on yield, chemical composition and ruminal degradability of cassava Hay in Southern Thailand

    Directory of Open Access Journals (Sweden)

    Pin Chanjula

    2007-01-01

    Full Text Available A 2x2 factorial arrangement in a randomized complete block design with 4 replications was carried out to investigate the effects of varieties (V, KU50 and RY72 and timing of subsequent cutting (SC, 2 and 3 months after cutting on yield and chemical composition of cassava hay. The results revealed that both total hay yield and crude protein dry matter yield were not significantly (P>0.05 influenced by variety or subsequent cutting. Cassava could produce from 4.98 to 6.15 t/ha of DM and 1.29 to 1.39 t/ha of CP. CP content in cassava plant ranged from 23.03 to 26.55% and was affected by different SC regimes. Their DM and CP rate of degradability of cassava hay (CH were studied using the nylon bag technique, in ruminally fistulated crossbred beef steers. The ruminal disappearance characteristics of CH and the effective degradability of DM and CP of all treatments were not significantly different (P>0.05 among the treatments. Based on this research, it was concluded that two varieties with subsequent cutting at 2 or 3 months intervals for 9 months was optimal to obtain high dry matter and protein yield as well as moderate-high nutritive value cassava hay. Moreover, it could be cultivated to produce CH with high nutritive value in southern conditions.

  3. Ruminal degradation kinetic of Brachiaria decumbens silages with different nitrogen additives

    Directory of Open Access Journals (Sweden)

    Odimári Pricila Pires do Prado

    2014-02-01

    Full Text Available This study aimed to assess levels of nitrogen additive Silogen® pasto on ruminal in vitro degradability gas production in Brachiaria decumbens silages. The nitrogen additive contained bacterial strains (Bacillus subtilis, Lactobacillus curvatus, Lactobacillus plantarum and Pediococcus acidilactici and minimum nitrogen to 360 g/kg. The levels were: 0.0; 1.0; 1.5; 2.0 and 2.5%. In fractionation were calculated total carbohydrates (TC, non-fiber carbohydrates (NFC, soluble and rapidly degradable (A + B1, potentially degradable (B2 and degradable (C. The kinetic parameters of FC and NFC were estimated using the technique of in vitro gas production. Data were subjected to analysis of variance at 5%. The level additive did not influence the TC and fraction A + B1. The levels of nitrogen additive influenced the NFC, fractions B2 and C. For NFC content 2.0% showed the lowest (22.0 %. The lower levels of additive nitrogen (0, 1.0 and 1.5 % had higher fractions of B2 (average 40.2 % and lower values for the fraction C (average 20.0 % . There was no difference in the volume of gas CNF (average 86.73 mL and final volume of gas produced (average 195.79 mL. Was no influence of nitrogen additives for the time of colonization, the lowest time of 3.89 h to 1.0%. The volume of gas of FC was influenced by levels of nitrogen additives with higher values to 0 % from 1.0 %, and 114.74 and 115.09 mL, respectively. Degradation rates of FC and FNC were also affected by the concentrations of nitrogenous additives, which presented higher rates to the levels of 2.0 and 2.5%. It follows that the lower levels of additives to 1.5 % promoted the reduction of the C fraction and increased B2, and greater production of gas volume of the fiber in these silages, showing better nutritional value in these silages.

  4. Efeito das fontes de amido e nitrogênio de diferentes degradabilidades ruminais.1. Digestibilidades parcial e total Effect of starch and nitrogen sources of different ruminal degradability. 1. Total and partial digestibility

    Directory of Open Access Journals (Sweden)

    Fábio Luiz Fregadolli

    2001-06-01

    factorial arrangement, were evaluated. Four Holstein steers (334 kg fitted with ruminal and duodenal cannulas, in a 4 x 4 Latin Square were used. Chromic oxide was used as a marker for drying matter flow. Interactions of starch and N sources on apparent digestibility coefficient (ADC of crude protein (CP and neutral detergent fiber (NDF was detected. Diets of corn and yeast, cassava hulls and cottonseed meal + meat and bone meal showed higher ADC of CP than diets of corn and meat and bone meal and with cassava hulls and yeast. The diet of starch and N with high ruminal degradability showed lower value than others diets, that was not different. Ruminal digestion of NDF was higher when starch source was corn compared with cassava hulls. The highest digestion of starch was higher in diets with cassava hulls than in diets of corn, in all segments of digestive system. Negative values for ruminal digestion of CP to experimental diets were observed. The N source influenced ruminal and intestinal digestion of energy. The ruminal digestion was higher in diets with yeast e intestinal digestion was higher in diets with cottonseed meal + meat and bone meal.

  5. Protein co-products and by-products of the biodiesel industry for ruminants feeding

    Directory of Open Access Journals (Sweden)

    Ricardo Andrés Botero Carrera

    2012-05-01

    Full Text Available The objective of the experiment was to classify 20 protein co-products and by-products of the biodiesel industry with potential to use in ruminant feeding. The meals evaluated were: cottonseed, canudo-de-pito, crambe, sunflower, castor-oil seeds detoxified with calcium, non-detoxified castor-oil seeds and soybean; and the cakes were: cottonseed, peanut, babassu, crambe, palm oil, sunflower, licuri, macauba seeds, non-detoxified castor-oil seeds, turnip and jatropha. The samples were quantified to determine dry matter (DM, organic matter (OM, crude protein (CP, ether extract (EE, neutral detergent fiber corrected for ash and protein (NDFap, non-fiber carbohydrates (NFC, acid detergent fiber corrected for ash and protein (ADFap, lignin, cutin and starch levels. The CP profile was characterized in fractions A, B1, B2, B3 and C. The in vitro dry matter digestibility (IVDMD, in vitro neutral detergent fiber digestibility (IVNDFD, rumen degradable and undegradable protein, intestinal digestibility, indigestible neutral detergent fiber and undegradable neutral detergent insoluble protein were evaluated. The OM, CP, EE, NDFap, NFC, ADFap, lignin, cutin and starch contents varied from 81.95 to 95.41%, 18.92 to 57.75%, 0.56 to 18.40%, 10.13 to 62.30%, 3.89 to 27.88%, 6.15 to 36.86%, 1.19 to 5.04%, 0 to 17.87% and 0.68 to 14.50%, respectively. The values of fractions A, B1, B2, B3 and C ranged from 5.40 to 43.31%, 0.08 to 37.63%, 16.75 to 79.39%, 1.86 to 59.15% and 0.60 to 11.47%, respectively. Concentrations of IVDMD, IVNDFD, rumen-degradable and undegradable protein, intestinal digestibility, indigestible NDF and undegradable neutral detergent insoluble protein ranged from 31.00 to 95.92%, 55.04 to 97.74%, 41.06 to 97.61%, 2.39 to 58.94, 9.27 to 94.26%, 1.05 to 40.80% and 0.29 to 2.92%, respectively. Some of these products can replace soybean meal, specially the Macauba seeds cake, cottonseed meal and peanut and turnip cakes based on digestive

  6. Soluble Non-ammonia Nitrogen in Ruminal and Omasal Digesta of Korean Native Steers Supplemented with Soluble Proteins

    Directory of Open Access Journals (Sweden)

    C. W. Choi

    2012-09-01

    Full Text Available An experiment was conducted to study the effect of soluble protein supplements on concentration of soluble non-ammonia nitrogen (SNAN in the liquid phase of ruminal (RD and omasal digesta (OD of Korean native steers, and to investigate diurnal pattern in SNAN concentration in RD and OD. Three ruminally cannulated Korean native steers in a 3×3 Latin square design consumed a basal diet of rice straw and corn-based concentrate (control, and that supplemented (kg/d DM basis with intact casein (0.24; IC or acid hydrolyzed casein (0.46; AHC. Ruminal digesta was sampled using a vacuum pump, whereas OD was collected using an omasal sampling system at 2.0 h intervals after a morning feeding. The SNAN fractions (free amino acid (AA, peptide and soluble protein in RD and OD were assessed using the ninhydrin assay. Concentrations of free AA and total SNAN in RD were significantly (p<0.05 lower than those in OD. Although free AA concentration was relatively high, mean peptide was quantitatively the most important fraction of total SNAN in both RD and OD, indicating that degradation of peptide to AA rather than hydrolysis of soluble protein to peptide or deamination may be the most limiting step in rumen proteolysis of Korean native steers. Diurnal variation in peptide concentration in OD for the soluble protein supplemented diets during the feeding cycle peaked 2 h post-feeding and decreased thereafter whereas that for the control was relatively constant during the entire feeding cycle. Diurnal variation in peptide concentration was rather similar between RD and OD.

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

    African Journals Online (AJOL)

    and the in situ digestion rate (Erdman, Vandersall, Russek-. Cohen & Switalski, 1987), it is imperative that these measurements be made in dairy cows at productive levels of intake. Values for UDP in published tables, e.g. those of. NRC (1989), should be used with caution in formulating diets for high-producing dairy cattle.

  8. In vitro evaluation of utilizable crude protein using ruminal fluid in ...

    African Journals Online (AJOL)

    The prime objective of this study was to evaluate the effective utilizable crude protein (uCP) in leaves, whole and seeds-removed pods of Moringa stenopetala and Moringa oleifera using ruminal fluid in vitro. Samples were analyzed for proximate nutrients using official methods. The metabolizable energy (ME), organic ...

  9. Estimation of ruminal and intestinal digestion profiles, hourly effective degradation ratio and potential N to energy synchronization of co-products from bioethanol processing.

    Science.gov (United States)

    Nuez-Ortín, Waldo G; Yu, Peiqiang

    2010-09-01

    Little research has been conducted to determine the magnitude of the differences in nutritive value among wheat dried distillers' grains with solubles (DDGS), corn DDGS and blend DDGS, or between different bioethanol plants. The objectives of this study were to compare different types of DDGS and different bioethanol plants in terms of: (1) rumen degradation kinetics profile of each DDGS component and rumen availability; (2) intestinal digestion profile of rumen undegraded protein; (3) hourly effective rumen degradation ratio and potential N-to-energy synchronization; (4) the role of acid detergent insoluble nitrogen in the determination of nutrient availability of DDGS. In addition, these parameters were compared in DDGS as opposed to parental grain. (1) The effective degradability of dry matter in DDGS samples increased as the content of feedstock wheat increased. DDGS are a good source of rumen-undegradable protein. The protein content of DDGS derived from wheat is higher relative to that derived from corn; however, the undegradability of the protein fraction increases as the proportion of corn in the feedstock augments. (2) In addition, DDGS provide significant amounts of rumen-degradable protein, which increased as the content of wheat in the feedstock increased. This indicates a potential loss of N when high levels of DDGS are included in the diet. (3) Acid detergent insoluble crude protein (ADICP) levels were low across DDGS samples, revealing no effect on ruminal and intestinal disappearance of protein. However, consideration should be given to the numerical differences in digestibility of rumen-undegradable protein and the relation to ADICP content. (4) Further research with a higher number of samples and higher variability in the ADICP content should be undertaken to investigate the effect of ADICP on rumen and intestinal disappearance of DDGS protein. The digestive characteristics of each DDGS component (dry matter, organic matter, crude protein and

  10. Considering ruminal interactions in energy and protein evaluation systems for ruminants in Denmark

    DEFF Research Database (Denmark)

    Madsen, J; Hvelplund, Torben; Weisbjerg, Martin Riis

    2007-01-01

    The transformation of the feeds in the rumen by microbes is included in most protein evaluation systems introduced in the 1980 and 1990's but it is only partly considered in present energy evaluation systems....

  11. Protection of sunflower seed and sunflower meal protein with malic acid and heat: effects on in vitro ruminal fermentation and methane production.

    Science.gov (United States)

    Vanegas, Jorge Leonardo; Carro, Maria Dolores; Alvir, Maria Remedios; González, Javier

    2017-01-01

    Combined malic acid-heat treatments of protein supplements have been shown to reduce ruminal protein degradation, but there is no information on their possible influence on ruminal fermentation and methane emissions. This study aimed to investigate the effects of the treatment of sunflower meal (SM) and sunflower seed (SS) with malic acid and subsequent drying at 150°C for 1 (MAL1) or 3 h (MAL3) on in vitro rumen fermentation and methane emission using ruminal fluid from sheep as inoculum. Compared with untreated samples, the MAL3 treatment reduced (P 0.05) total volatile fatty acid production for any feed. This treatment also increased (P methane production (by 15.5% and 11.3%, respectively) and ammonia-N concentrations (by 26.5% and 14.5%, respectively). The MAL1 treatment was effective in reducing both ammonia-N concentrations and methane emissions without depressing SS and SM fermentation, but more research is needed to formulate environmentally cleaner diets for ruminants. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  12. Effect of Electron Beam Irradiation on Degradability Coefficients and Ruminalpostruminal Digestibility of Dry Matter and Crude Protein of some Plant Protein Sources

    Directory of Open Access Journals (Sweden)

    gasem tahan

    2016-06-01

    Full Text Available Effect of electron beam irradiation on degradability coefficients and ruminal- postruminal digestibility of dry matter and crude protein of soybean meal, canola meal and Lathyrus sativus seed, irradiated at doses of 50, 100 and 150 kGy was investigated. Ruminal degradability of dry matter and crude protein was determined by in situ method using two cannulated Holstein heifers. Ruminal- postruminal digestibility of dry matter and crude protein was determined by in situ (nylon bag-in vitro (daisy digestor techniques. Data analyzed using SAS software as randomized completely design and the treatment means were compared using Tukey test. The results indicated that irradiation had no effect on dry matter, ether extract and ash content of feeds. In soybean meal, washout fraction and potentially degradable fraction of dry matter and crude protein was higher and lower at dose of 150 kGy irradiation than other treatments, respectively, and degradation rate constant and ruminal effective degradability of dry matter and crude protein was lower at all doses of irradiation than untreated soybean meal. In canola meal, irradiation at doses of 50 and 100 kGy decreased washout fraction and increased potentially degradable fraction of crude protein compared with untreated canola meal. In Lathyrus sativus seed, only potentially degradable fraction of dry matter and crude protein was lower at dose of 150 kGy irradiation than untreated Lathyrus sativus seed. Ruminal digestibility of crude protein decreased in soybean meal at doses of 100 and 150 kGy irradiation and for canola meal at all doses of irradiation than untreated samples. Total tract digestibility of crude protein decreased in soybean meal at dose of 150 kGy irradiation and for canola meal at all doses of irradiation than untreated samples. In Lathyrus sativus seed, ruminal-postruminal digestibility and total tract digestibility of dry matter increased at doses of 100 and 150 kGy irradiation than untreated

  13. Proteins Profile in Milk from Three Species of Ruminants

    OpenAIRE

    Cozma, Anamaria; Sanda ANDREI; Miere, Doina; Filip, Lorena; Felicia LOGHIN

    2011-01-01

    Milk proteins, caseins and whey proteins, are very important nutritionally, as they contain all essential aminoacids in optimal proportions and are the most important source of bioactive peptides. These peptides are protein fragments resulting from enzymatic hydrolysis of milk proteins, which carry numerous beneficial effects on the cardiovascular, nervous, gastrointestinal and immune systems. In this research, total proteins, caseins and whey proteins respectively, were dosed in milk from th...

  14. Identification of amino-acid residues in the V protein of peste des petits ruminants essential for interference and suppression of STAT-mediated interferon signaling

    Energy Technology Data Exchange (ETDEWEB)

    Ma, Xusheng, E-mail: maxushengtt@163.com [State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Lanzhou 730030 (China); Yang, Xing [State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Lanzhou 730030 (China); Nian, Xiaofeng [Institute of Pathogen Biology and Immunology, Hebei North University, Zhangjiakou 07500 (China); Zhang, Zhidong; Dou, Yongxi [State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Lanzhou 730030 (China); Zhang, Xuehu [Gansu Agricultural University, Lanzhou (China); Luo, Xuenong; Su, Junhong; Zhu, Qiyun [State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Lanzhou 730030 (China); Cai, Xuepeng, E-mail: caixp@vip.163.com [State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Lanzhou 730030 (China)

    2015-09-15

    Peste des petits ruminants virus (PPRV) causes a fatal disease in small ruminants. V protein of PPRV plays a pivotal role in interfering with host innate immunity by blocking IFNs signaling through interacting with STAT1 and STAT2. In the present study, the results demonstrated that PPRV V protein blocks IFN actions in a dose dependent manner and restrains the translocation of STAT1/2 proteins. We speculate that the translocation inhibition might be caused by the interfering of the downstream of STAT protein. Mutagenesis defines that Cys cluster and Trp motif of PPRV V protein are essential for STAT-mediated IFN signaling. These findings give a new sight for the further studies to understand the delicate mechanism of PPRV to escape the IFN signaling. - Highlights: • PPRV V protein inhibits type I IFN production and blocks its activation. • PPRV V protein negatively regulates activation of ISRE and GAS promoter. • PPRV V protein inhibits nuclear translocation of STAT protein by non-degradation. • PNT and VCT domain of PPRV V protein inhibit IFN transduction. • PPRV V protein binds with STAT protein via some conserved motifs.

  15. Degradação ruminal da silagem de milho e da palha de arroz utilizando enzimas fibrolíticas exógenas = Ruminal degradation of corn silage and rice straw using exogenous fibrolityc enzymes

    Directory of Open Access Journals (Sweden)

    Adriana de Souza Martins

    2008-10-01

    Full Text Available Estudou-se o efeito da adição de enzimas fibrolíticas (celulase e xilanase sobre a degradabilidade in situ da matéria seca (MS, proteína bruta (PB, fibra em detergente neutro (FDN, fibra em detergente ácido (FDA e hemicelulose da silagem de milho (SMe da palha de arroz (PA, utilizando seis bovinos fistulados no rúmen. Também foi avaliada a proporção de nitrogênio (N insolúvel em detergente ácido em relação ao N total (NIDA/N total. As enzimas fibrolíticas foram extraídas dos fungos Aspergillus niger eTrichoderma longibrachiatum, sendo fornecido 0,75 g kg-1 de MS animal-1 dia-1, via cânula ruminal. Os tempos de incubação foram: 0, 3, 6, 12, 24, 48, 72 e 96 h. Os resíduos de incubação foram comparados por meio da microscopia eletrônica de varredura (MEV. Afração solúvel da MS e da PB da silagem de milho aumentou com a adição de enzimas fibrolíticas. Não houve efeito das enzimas sobre a degradação da parede celular dos volumosos. A adição de enzimas não alterou o teor de NIDA/N total da silagem de milho, porém aumentou esta proporção nos resíduos da palha de arroz incubados durante 12 e 48 h. As observações ao MEV indicaram aumento da colonização bacteriana sobre a parede celular dos volumosos, mas sem efeito sobre a degradação ruminal.This study evaluated the effect of fibrolytic enzyme addition(cellulase and xylanase on the dry matter (DM, crude protein (CP, neutral detergent fiber (NDF, acid detergent fiber (ADF and hemicellulose in situ degradability of corn silage (CS and rice straw (RS, using six cattle cannulated on the rumen. Acid detergentinsoluble nitrogen (ADIN/total N was evaluated. The fibrolytic enzymes were extracted from Aspergillus niger and Trichoderma longibrachiatum fungi, supplied at 0.75 g kg-1 of DM animal-1 day-1, through ruminal cannula. The incubation times were 0, 3, 6, 12, 24, 48, 72 and 96 hours. The incubation residues were compared by scanning electron micrograph (SEM

  16. CHEMICAL COMPOSITION, DEGRADABILITY AND METHANE EMISSION POTENTIAL OF BANANA CROP RESIDUES FOR RUMINANTS

    Directory of Open Access Journals (Sweden)

    Lincoln Nunes Oliveira

    2014-08-01

    Full Text Available Banana leaf hay (BL, banana pseudostem hay (BP, coast-cross hay (CC, 50% coast-cross hay with 50% banana leaf (BLCC and 50% coast-cross hay with 50% pseudostem hay (BPCC were evaluated for chemical composition, cumulative gas production, dry matter degradability and methane emission potential. Inoculums from sheep and cattle were collected to tests. The experimental design was completely randomized in a factorial arrangement of 5 x 2, being data analysed by variance analysis and the means compared by Tukey test (5%. The crude protein levels for the substrates BL, BP, CC, BLCC and BPCC were respectively 13.8%, 3.5%, 8.6%, 9.7% and 6.1%. Despite its low protein level, the BP substrate had higher content of non-fibrous carbohydrates (28.4%, followed by BL (23.4%, BPCC (23.4%, BLCC (20.0% and CC (13.3%. The highest cumulative gas production was observed for BP (P < 0.05, reflecting their greater effective degradability (76.3%. This substrate showed the largest emissions of methane (34,16 mL/g DMD.

  17. Prevention of leucaena toxicosis of cattle in Florida by ruminal inoculation with 3-hydroxy-4-(1H)-pyridone-degrading bacteria.

    Science.gov (United States)

    Hammond, A C; Allison, M J; Williams, M J; Prine, G M; Bates, D B

    1989-12-01

    Ruminal microorganisms in cattle at a Florida agriculture research station did not have the ability to detoxify leucaena by degradation of 3-hydroxy-4(1H)-pyridone (3,4,-DHP), but a DHP isomer (2,3-DHP) was degraded in some cattle. Cattle with microorganisms that degraded 2,3-DHP were mostly Senepol cattle imported from St. Croix, US Virgin Islands, where leucaena is an indigenous species. Hereford cattle at the research station in Florida generally did not degrade 3,4-DHP or 2,3-DHP. An experiment was conducted in which a pure culture of 3,4-DHP-degrading bacteria was inoculated into Hereford cattle (with ruminal fistula) grazing leucaena. The bacteria successfully colonized the rumen of recipient cattle and persisted through the following winter when there was no leucaena in the diet.

  18. 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

  19. Nutrient Content, Protein Fractionation, and Utilization of Some Beans as Potential Alternatives to Soybean for Ruminant Feeding

    Directory of Open Access Journals (Sweden)

    A. Jayanegara

    2016-12-01

    Full Text Available This experiment aimed to determine nutrient content, protein fraction, and in vitro rumen fermentation of some alternative beans in comparison to soybean. Samples used were napier grass, soybean, redbean, groundnut, pigeonpea, cowpea, bambarabean, and mungbean. Samples were determined for their proximate composition, Van Soest’s fiber fraction, and Cornell protein fraction. The samples were subsequently evaluated for their fermentation characteristics and digestibility by using a two-stage in vitro rumen fermentation technique, maintained at 39 oC for 2 × 48 h. The in vitro incubation was performed in three consecutive runs by following a randomized complete block design in which each sample per run was represented by four fermentation tubes. Results revealed that all experimental beans contained high crude protein (CP, i.e. above 200 g/kg dry matter (DM, but only soybean and groundnut had CP contents higher than 300 g/kg DM. Redbean had the lowest crude fiber and acid detergent fiber contents among the beans. Soybean contained high proportion of rapidly degraded CP fraction, but low in slowly degraded and unavailable CP fractions. High proportion of slowly degraded CP fraction was found in redbean and bambarabean. Redbean, pigeonpea, cowpea, and mungbean were better than soybean, groundnut, and bambarabean with regard to DM degradability and DM digestibility values (P<0.05. Concentration of total VFA was the highest in the incubation of redbean. It was concluded that groundnut, redbean, pigeonpea, cowpea, and mungbean have the potency to be used to substitute soybean for ruminant feeding.

  20. Degradação ruminal da silagem de capim-elefante aditivado com cana-de-açúcar e farelo de cacau Ruminal degradation of elephant grass sillage added with sugarcane and cocoa meal

    Directory of Open Access Journals (Sweden)

    Fábio Andrade Teixeira

    2008-06-01

    Full Text Available Objetivou-se avaliar a degradação ruminal da matéria seca (MS, da proteína bruta (PB e da fibra em detergente neutro (FDN das silagens de capim-elefante; capim-elefante ensilado com 30% de cana-de-açúcar; 15% de farelo de cacau; e 45% de cana mais 15% de farelo de cacau. Amostras de cada silagem foram incubadas no rúmen de três novilhos mestiços holandês x zebu durante os períodos de 0, 12, 24, 48, 72 e 96 horas. O delineamento experimental utilizado foi o de parcelas subdivididas e os coeficientes não lineares a, b e c foram estimados por meio de procedimentos iterativos de Gauss-Newton. Para o capim-elefante ensilado com farelo de cacau, foram observados maiores valores da fração a da MS e da PB, próximos a 60%, além de maiores taxas da degradação da fração b e da degradabilidade efetiva (DE da PB para as taxas de passagem 2, 5 e 8% h-1. As silagens que apresentaram as maiores frações não-degradáveis i da FDN foram as silagens de capim-elefante sem aditivo e com 30% de cana, enquanto a silagem aditivada com 45% de cana e 15% de farelo de cacau destacou-se pela maior degradação potencial da MS, da PB e da FDN, em todos os períodos estudados, e maiores DE da MS e da PB para as taxas de passagem 2, 5 e 8% h-1. A inclusão associada da cana-de-açúcar com o farelo de cacau, no momento da ensilagem do capim-elefante, apresenta-se como alternativa para aumentar a degradabilidade da forragem em estádio de crescimento avançado.The objective was to evaluate the dry matter (DM, crude protein (CP and neutral detergent fiber (NDF ruminal degradation of silages of elephant grass; elephant grass ensiled with 30% of sugarcane; 15% of cocoa meal; and 45% of sugarcane plus 15% of cocoa meal. Samples of each silage were incubated in the rumen of three Holstein x Zebu crossbred steers during the periods of 0, 12, 24, 48, 72 and 96 hours. The experimental design used was the one of subdivided parcels and the non linear coefficients a

  1. Degradação ruminal de silagem de capim-elefante com adição de vagem de algaroba triturada Ruminal degradation of elephant grass silage with mesquite pods

    Directory of Open Access Journals (Sweden)

    Aníbal Coutinho do Rêgo

    2011-03-01

    Full Text Available Esta pesquisa foi realizada visando-se avaliar a degradação ruminal da matéria seca (MS, proteína bruta (PB e fibra em detergente neutro (FDN de silagens de capim-elefante colhido aos 70; 90 e 110 dias após rebrota, com inclusão de 0; 5; 10 e 15% de vagem de algaroba triturada, com base na matéria natural, em delineamento inteiramente casualizado arranjado em parcelas subdivididas. Amostras de cada silagem foram incubadas no rúmen de duas vacas Jersey por 3; 6; 12; 24; 48; 72 e 96 h, sendo os saquinhos referentes ao tempo zero apenas lavados em água para determinação da fração solúvel. Não houve interação (P > 0,05 tempo de incubação x inclusão de vagem de algaroba x idade de corte para degradabilidade da MS, embora tenha ocorrido interação destes fatores para degradabilidade da PB e FDN. A maior degradabilidade efetiva (DE da MS (42,54% foi observada para 15% de inclusão de vagem de algaroba. A DE da PB foi maior (69,04% para silagem de capim-elefante com 70 dias de idade com 15% de vagem de algaroba. A inclusão de vagem de algaroba triturada à silagem de capim-elefante melhora a degradabilidade da MS, PB e FDN, enquanto o avanço da idade após rebrota resulta em redução destes parâmetros.This research was carried out to evaluate the ruminal degradation of dry matter (DM, crude protein (CP and neutral detergent fiber (NDF of silages of elephant grass (Pennisetum purpureum cutting in 70; 90 and 110 days after regrowth with inclusion of 0; 5; 10 and 15% of mesquite (Prosopis juliflora meal, based on natural matter in a completely randomized design, in split plot arrangement. Samples of silages were incubated in the rumen of two Jersey cows for 3; 6; 12; 24; 48; 72 and 96 h, and the bags at time "zero" were only washed with water to determine the soluble fraction. There was not interaction (P > 0.05 incubation time × inclusion of mesquite pods × cutting age of the grass for DM degradability, there was only

  2. Ruminal and postruminal digestion of dietary protein and starch in ...

    African Journals Online (AJOL)

    diet per se did not affect the amount of NAN passing to the duodenum, but its interaction with OMDR was highly significant. Thus, with lower .... RDP, effected by the following protein supplements: I .44% urea ( I 17 g CPkg DM; 7 4oh ..... tract shows the opposite trend to that of starch, although the relationship is not close, ...

  3. Recombinant capripoxviruses expressing proteins of bluetongue virus: evaluation of immune responses and protection in small ruminants.

    Science.gov (United States)

    Perrin, Aurélie; Albina, Emmanuel; Bréard, Emmanuel; Sailleau, Corinne; Promé, Sylvie; Grillet, Colette; Kwiatek, Olivier; Russo, Pierre; Thiéry, Richard; Zientara, Stephan; Cêtre-Sossah, Catherine

    2007-09-17

    The development of recombinant capripoxviruses for protective immunization of ruminants against bluetongue virus (BTV) infection is described. Sheep (n=11) and goats (n=4) were immunized with BTV recombinant capripoxviruses (BTV-Cpox) individually expressing four different genes encoding two capsid proteins (VP2 and VP7) and two non-structural proteins (NS1, NS3) of BTV serotype 2 (BTV-2). Seroconversion was observed against NS3, VP7 and VP2 in both species and a lymphoproliferation specific to BTV antigens was also demonstrated in goats. Finally, partial protection of sheep challenged 3 weeks after BTV-Cpox administration with a virulent strain of BTV-2, was observed.

  4. Restoration of in situ fiber degradation and the role of fibrolytic microbes and ruminal pH in cows fed grain-rich diets transiently or continuously.

    Science.gov (United States)

    Pourazad, P; Khiaosa-Ard, R; Metzler-Zebeli, B U; Klevenhusen, F; Zebeli, Q

    2017-12-01

    In this study, we used two different grain-rich feeding models (continuous or transient) to determine their effects on in situ fiber degradation and abundances of important rumen fibrolytic microbes in the rumen. The role of the magnitude of ruminal pH drop during grain feeding in the fiber degradation was also determined. The study was performed in eight rumen-fistulated dry cows. They were fed forage-only diet (baseline), and then challenged with a 60% concentrate diet for 4 weeks, either continuously (n=4 cows) or transiently (n=4 cows). The cows of transient feeding had 1 week off concentrate in between. Ruminal degradation of grass silage and fiber-rich hay was determined by the in situ technique, and microbial abundances attached to incubated samples were analyzed by quantitative PCR. The in situ trials were performed at the baseline and in the 1st and the last week of concentrate feeding in the continuous model. The in situ trials were done in cows of the transient model at the baseline and in the 1st week of the re-challenge with concentrate. In situ degradation of NDF and ADF of the forage samples, and microbial abundances were determined at 0, 4, 8, 24 and 48 h of the incubation. Ruminal pH and temperature during the incubation were recorded using indwelling pH sensors. Compared with the respective baseline, both grain-rich feeding models lowered ruminal pH and increased the duration of pH below 5.5 and 5.8. Results of the grass silage incubation showed that in the continuous model the extent of NDF and ADF degradation was lower in the 1st, but not in the last week compared with the baseline. For the transient model, degradation of NDF of the silage was lower during the re-challenge compared with the baseline. Degradation of NDF and ADF of the hay was suppressed by both feeding models compared with the respective baseline. Changes in fiber degradation of either grass silage or hay were not related to the magnitude of ruminal pH depression during grain

  5. Protein misfolding and degradation in genetic diseases

    DEFF Research Database (Denmark)

    Bross, P; Corydon, T J; Andresen, B S

    1999-01-01

    Investigations of genetic diseases such as cystic fibrosis, alpha-1-antitrypsin deficiency, phenylketonuria, mitochondrial acyl-CoA dehydrogenase deficiencies, and many others have shown that enhanced proteolytic degradation of mutant proteins is a common molecular pathological mechanism. Detailed....... This appears to be the case for many missense mutations and short in-frame deletions or insertions that represent a major fraction of the mutations detected in genetic diseases. In some diseases, or under some circumstances, the degradation system is not efficient. Instead, aberrant folding leads...... in which proteins fold. Within the context of genetic diseases, we review knowledge on the molecular processes underlying protein quality control in the various subcellular compartments. The important impact of such systems for variability of the expression of genetic deficiencies is emphasised....

  6. Evaluation of Camelina sativa (L.) Crantz meal as an alternative protein source in ruminant rations.

    Science.gov (United States)

    Colombini, Stefania; Broderick, Glen A; Galasso, Incoronata; Martinelli, Tommaso; Rapetti, Luca; Russo, Roberto; Reggiani, Remo

    2014-03-15

    Camelina sativa (CS) is an oilseed crop used for biofuel production. By-products from oil extraction are high in protein and can be used in ruminant rations; more information about their nutritive value is required also considering the antinutrional factor content of the by-products. The aim of this study was to evaluate the nutritive value of CS meal genotypes in comparison with canola. Ten CS genotypes and one canola cultivar were evaluated. Meals were obtained from seeds after solvent oil extraction. CS average crude protein (CP) content (g kg⁻¹ dry matter) was 457. Numerical differences in lysine and sulfur amino acid content were observed among CS genotypes. Glucosinolate (mmol kg⁻¹) content was higher for CS (23.1) than canola (7.2). Sinapine content (g kg⁻¹) was lower for CS (2.79) than for canola (4.32). Differences were observed among CS genotypes for rumen undegraded protein (RUP). Average RUP (g kg⁻¹ CP) was 316 for CS and 275 for canola. CS meal has potential for use in ruminant rations as a high-quality protein source. In vivo studies are needed to compare CS with other protein sources used in cattle rations. Implementation of breeding programs for improved meal quality is recommend. © 2013 Society of Chemical Industry.

  7. Ruminal and postruminal digestion of dietary protein and starch in ...

    African Journals Online (AJOL)

    glucose available for absorption in the small intestine. ln this regard, the effect of protein concentration and deg- radation, and energy .... 20%cP. 88 3.9. 89. Maize gluten,. 60%cP. 93 ll. 32. Maize meal. 89 1.8 75. Sorghum meal 89 L6. 69. Wheat bran. 89 2.8. 92. 60 2.0 3.7 30 35. 5. r. 9.1 8.8 4.1 65 85 12.9. 34. 25. 1 3 l 0 t 3.

  8. Ruminal tryptophan-utilizing bacteria degrade ergovaline from tall fescue seed extract

    Science.gov (United States)

    The objectives of this study were to evaluate degradation of ergovaline in a tall fescue [Lolium arundinaceum (Schreb.) Darbysh.] seed extract by rumen microflora ex vivo and to identify specific bacteria capable of ergovaline degradation in vitro. Rumen cell suspensions were prepared by harvesting ...

  9. Degradation of amyloid beta protein by purified myelin basic protein.

    Science.gov (United States)

    Liao, Mei-Chen; Ahmed, Mahiuddin; Smith, Steven O; Van Nostrand, William E

    2009-10-16

    The progressive accumulation of beta-amyloid (Abeta) in senile plaques and in the cerebral vasculature is the hallmark of Alzheimer disease and related disorders. Impaired clearance of Abeta from the brain likely contributes to the prevalent sporadic form of Alzheimer disease. Several major pathways for Abeta clearance include receptor-mediated cellular uptake, blood-brain barrier transport, and direct proteolytic degradation. Myelin basic protein (MBP) is the major structural protein component of myelin and plays a functional role in the formation and maintenance of the myelin sheath. MBP possesses endogenous serine proteinase activity and can undergo autocatalytic cleavage liberating distinct fragments. Recently, we showed that MBP binds Abeta and inhibits Abeta fibril formation (Hoos, M. D., Ahmed, M., Smith, S. O., and Van Nostrand, W. E. (2007) J. Biol. Chem. 282, 9952-9961; Hoos, M. D., Ahmed, M., Smith, S. O., and Van Nostrand, W. E. (2009) Biochemistry 48, 4720-4727). Here we show that Abeta40 and Abeta42 peptides are degraded by purified human brain MBP and recombinant human MBP, but not an MBP fragment that lacks autolytic activity. MBP-mediated Abeta degradation is inhibited by serine proteinase inhibitors. Similarly, Cos-1 cells expressing MBP degrade exogenous Abeta40 and Abeta42. In addition, we demonstrate that purified MBP also degrades assembled fibrillar Abeta in vitro. Mass spectrometry analysis identified distinct degradation products generated from Abeta digestion by MBP. Lastly, we demonstrate in situ that purified MBP can degrade parenchymal amyloid plaques as well as cerebral vascular amyloid that form in brain tissue of Abeta precursor protein transgenic mice. Together, these findings indicate that purified MBP possesses Abeta degrading activity in vitro.

  10. Mammalian 26S proteasomes remain intact during protein degradation

    DEFF Research Database (Denmark)

    Kriegenburg, Franziska; Seeger, Michael; Saeki, Yasushi

    2008-01-01

    It has been suggested that degradation of polyubiquitylated proteins is coupled to dissociation of 26S proteasomes. In contrast, using several independent types of experiments, we find that mammalian proteasomes can degrade polyubiquitylated proteins without disassembling. Thus, immobilized, (35)...

  11. Seminal-type ribonuclease genes in ruminants, sequence conservation without protein expression?

    Science.gov (United States)

    Kleineidam, R G; Jekel, P A; Beintema, J J; Situmorang, P

    1999-04-29

    Bovine seminal ribonuclease (BS-RNase) is an interesting enzyme both for functional and structural reasons. The enzyme is the product of a gene duplication that occurred in an ancestral ruminant. It is possible to demonstrate the presence of seminal-type genes in all other investigated ruminant species, but they are not expressed and show features of pseudogenes. In this paper we report the determination of two pancreatic and one seminal-type ribonuclease gene sequences of swamp-type water buffalo (Bubalus bubalis). The two pancreatic sequences encode proteins with identical amino acid sequences as previously determined for the enzymes isolated from swamp-type and river-type water buffalo, respectively. The seminal-type sequence has no pseudogene features and codes for an enzyme with no unusual features compared with the active bovine enzyme, except for the replacement of one of the cysteines which takes part in the two intersubunit disulfide bridges. However, Western blotting demonstrates the presence of only small amounts of the pancreatic enzymes in water buffalo semen, suggesting that also in this species the seminal-type sequence is not expressed. But it is still possible that the gene is expressed somewhere else in the body or during development. Reconstruction of seminal-type ribonuclease sequences in ancestors of Bovinae and Bovidae indicates no serious abnormalities in the encoded proteins and leads us to the hypothesis that the ruminant seminal-type ribonuclease gene has not come to expression during most of its evolutionary history, but did not exhibit a high evolutionary rate that is generally observed in pseudogenes.

  12. STUDY OF WHEAT PROTEIN DEGRADATION DURING GERMINATION

    Directory of Open Access Journals (Sweden)

    Eva Michalcová

    2012-06-01

    Full Text Available Glutens, the storage proteins of cereals, play significant role in technological and nutraceutical quality of cereal grains. Whereas the high content of glutens allows making better structure of dough, consummation of foods with high gluten content can cause digestive problems. In our work, we studied ability of wheat proteases to degrade proteins, especially glutens. Wheat grains were germinated for up to seven days at three different temperatures 15, 20, 30 °C and pH 3.0, 4.0, 5.5, 7.0, 8.0. Proteins were fractionated into salt-soluble albumins and globulins, alcohol-soluble gliadins and base-soluble glutenins. In these fractions, protein content and their composition were analyzed by Bradford method and SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis. The results showed that remarkable degradation of glutens started after three days and the lowest concentration was measured at the seventh day of germination of wheat grain at temperature 20 °C, pH 5.5.

  13. Heat Stress Alters Ruminal Fermentation and Digesta Characteristics, and Behavior in Lactating Dairy Cattle

    Science.gov (United States)

    In a study designed to assess the impact and interaction of nonfiber carbohydrates (NFC) and ruminally degradable protein (RDP) on ruminal characteristics and animal behavior, animals experienced heat stress in the first period (HS), and no/greatly reduced heat stress (NHS) in the second period, all...

  14. Ruminant Nutrition Symposium: The utility of lipid extracted algae as a protein source in forage or starch-based ruminant diets.

    Science.gov (United States)

    Lodge-Ivey, S L; Tracey, L N; Salazar, A

    2014-04-01

    (P < 0.01). Microbial efficiency results for Chlorella were more variable for Nannochloropsis with 1 Chlorella spp. increasing microbial efficiency by 36% over SOY (P < 0.05) and the other Chlorella spp. decreasing microbial efficiency by approximately 42% compared with SOY (P < 0.01). Overall, the results from both experiments are promising for LEA as a protein feedstuff in ruminant diets. Further research is necessary to fully understand the interactions and consequences of upstream processes and what role algal strain plays in LEA quality.

  15. Grain filling, starch degradation and feeding value of maize for ruminants

    NARCIS (Netherlands)

    Ali, M.

    2013-01-01

    Keywords; Maize (Zea mays L), Genotypes, Grain filling, Growth temperature, Kernels, Gas production, Starch degradation, Oven-drying, Silage, Ensiling temperature, Ensiling duration, Feeding value, Lactating cows Maize (Zea mays L.) is a major component in the ration of dairy cows in many parts of

  16. Improving ruminal degradability of oil palm fronds using white rot fungi

    NARCIS (Netherlands)

    Rahman, M.M.; Lourenco, M.; Hassim, H.A.; Baars, J.J.P.; Sonnenberg, A.S.M.; Cone, J.W.; Boever, de J.L.; Fievez, V.

    2011-01-01

    The use of oil palm fronds (OPF) in livestock production is limited as up to 0.20 of their dry biomass is lignin. White rot fungi (WRF) are very effective basidiomycetes for biological pre-treatment as they degrade lignin extensively. Ten WRF were screened for their potential to increase OPF

  17. Serum acute phase proteins in cows with SARA (Subacute Ruminal Acidosis suspect

    Directory of Open Access Journals (Sweden)

    C. Cannizzo

    2012-02-01

    Full Text Available The aim of this study was to evaluate the variations of Acute Phase Proteins (APPs and other blood constituents during the onset of the sub-acute ruminal acidosis (SARA pathological status. A total of 108 cows from 12 dairy herds were randomly selected and divided into three Groups of 36 animals each. All animals were subjected to a rumenocentesis. Group A was composed by subjects with a rumen pH>5.8, Group B was composed by subjects with a rumen pH ≤5.5≤5.8 and Group C was composed by subjects with a rumen pH<5.5. Blood samples were collected by jugular venipuncture and Haptoglobin (Hp, Serum Amyloid A (SAA, Total Proteins, Albumin and White Blood Cells (WBC were determined. One-way ANOVA showed a statistical significance on Rumen pH, Hp, SAA. SARA seems not stimulate the APPs production from liver.

  18. 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.

  19. Grain filling, starch degradation and feeding value of maize for ruminants

    OpenAIRE

    Ali, M.

    2013-01-01

    Keywords; Maize (Zea mays L), Genotypes, Grain filling, Growth temperature, Kernels, Gas production, Starch degradation, Oven-drying, Silage, Ensiling temperature, Ensiling duration, Feeding value, Lactating cows Maize (Zea mays L.) is a major component in the ration of dairy cows in many parts of the world. The currently increasing economic importance of maize has highlighted the need to determine its nutritional value, and to assess the factors influencing its nutritive value. Genotypic mak...

  20. Comparison of biuret and refractometry methods for the serum total proteins measurement in ruminants.

    Science.gov (United States)

    Katsoulos, Panagiotis D; Athanasiou, Labrini V; Karatzia, Maria A; Giadinis, Nektarios; Karatzias, Harilaos; Boscos, Constantin; Polizopoulou, Zoe S

    2017-12-01

    Determination of serum total protein concentration is commonly performed by the biuret method. Refractometric measurement is a faster and less expensive alternative but its accuracy has not been determined in ruminants. The purpose of the study was to compare the serum total protein concentrations in cattle, sheep, and goats measured by the biuret method with those obtained by refractometry. Serum total protein concentration was determined in 120 cattle, 67 sheep, and 58 goat blood samples refractometrically and with the biuret method. The data were analyzed with a paired samples t-test, and Passing and Bablok regression equations and Bland and Altman plots were generated. There was a strong linear relationship between the total protein values determined with the refractometer and the biuret method in cattle, sheep, and goats. The statistical accuracy, which represents a bias correction factor that measures the deviation of the best-fit line from the 45° line through the origin, was 90.63% for cattle, 93.05% for sheep, and 91.76% for goats. The mean protein values determined with the refractometer were significantly lower than those measured with the biuret method in cattle and goats (P  .05). The evaluated refractometer was sufficiently accurate for the determination of serum total proteins in cattle, sheep, and goats, although it cannot be used interchangeably with the biuret method. The RIs should be corrected for negative bias based on the created equations. © 2017 American Society for Veterinary Clinical Pathology.

  1. Ileal and faecal protein digestibility measurement in humans and other non-ruminants - a comparative species view

    NARCIS (Netherlands)

    Hendriks, W.H.; Baal, van J.; Bosch, G.

    2012-01-01

    A comparative non-ruminant species view of the contribution of the large intestinal metabolism to inaccuracies in nitrogen and amino acid absorption measurements is provided to assess potential implications for the determination of crude protein/amino acid digestibility in adult humans consuming

  2. Evaluation of ruminal degradation profiles of forages using bags made from different textiles

    OpenAIRE

    Tiago Neves Pereira Valente; Edenio Detmann; Augusto César de Queiroz; Sebastião de Campos Valadares Filho; Daiany Iris Gomes; Janderson Florêncio Figueiras

    2011-01-01

    The objective of this study was to evaluate the in situ degradation profiles of dry matter (DM) and neutral detergent fiber (NDF) of different forages using nylon (50 µm), F57 (Ankom®) and non-woven textile (NWT - 100 g/m²) bags. Eight forage samples were used: sugarcane, corn silage, elephant grass cut at 50 and 250 days of regrowth, corn straw, signal grass hay, coast cross hay, and fresh alfalfa. Samples were incubated for 0, 3, 6, 12, 18, 24, 48, 72, 96, 120, 144, 168, 192, 216, 240, and ...

  3. Substituição do Milho pela Farinha de Mandioca de Varredura em Dietas de Cabras em Lactação: Fermentação Ruminal e Concentrações de Uréia Plasmática e no Leite Replacement of Corn by Cassava By-Product Meal in the Lactating Goat Diets: Effects on Diet Degradability, Ruminal Fermentation and Plasma and Milk Urea Concentrations

    Directory of Open Access Journals (Sweden)

    Gisele Fernanda Mouro

    2002-07-01

    Full Text Available O objetivo deste trabalho foi avaliar o efeito da substituição do milho pela farinha de mandioca de varredura, em dietas de cabras Saanen em lactação, sobre a degradabilidade potencial, efetiva e efetiva corrigida das rações, bem como sobre o pH ruminal e as concentrações de amônia ruminal, uréia plasmática e uréia no leite em cabras em lactação. Foram utilizadas quatro cabras há 100 dias em lactação. O delineamento utilizado foi o quadrado latino 4 x4, em que os tratamentos consistiram em níveis de 0, 33, 67 e 100% de substituição do milho pela farinha de mandiocade varredura. A degradabilidade das rações experimentais foi determinada em bovinos pela técnica in situ. Apesar de a degradabilidade potencial das dietas experimentais ter tido pequena variação para MS, PB e amido, a degradabilidade efetiva e a degradabilidade efetiva corrigida da MS, da PB e do amido aumentaram com a substituição do milho pela farinha de varredura de mandioca. Os tratamentos não influenciaram o pH ruminal, bem como as concentrações de NH3-ruminal, uréia plasmática e uréia do leite. Houve correlação positiva (PThe objectives of this work were to evaluate effects of replacing corn by cassava by-product meal, in diets of Saanen lactating goats, on potencial, effective and corrected effective degradability of diets with steers, and on ruminal pH and ruminal ammonia, plasma urea nitrogen (PUN and milk urea nitrogen (MUN concentrations in lactating goats. Two steers and four multiparous goats fitted with ruminal cannula and 100 days in milking were used. The design was a 4 x 4 Latin square and treatments as following: 0, 33, 67 and 100% replacement of corn by cassava by-product meal. Rations degradability was determined using in situ technique. Potential degradability of experimental diets showed small variation for dry matter (DM, crude protein (CP and starch (S but effective and corrected effective degradability increased as corn was

  4. Degradação da matéria seca e da proteína bruta de silagens de milho sem espigas com cana-de-açúcar e bagaço de mandioca - DOI: 10.4025/actascianimsci.v28i4.605 Dry mater and crude protein ruminal degradation of corn silage without spikes with sugar cane and manioc pulp - DOI: 10.4025/actascianimsci.v28i4.605

    Directory of Open Access Journals (Sweden)

    Paulo Rossi Junior

    2006-02-01

    Full Text Available A degradação ruminal da matéria seca (MS e da proteína bruta (PB de silagen s de milho sem espigas (SMSE com cana -de-açúcar (CA e bagaço seco de mandioca (BSM, comparadas com a silagem de milho com espigas (SMCE, foi avaliada. A ensilagem foi feita com (CI e sem inoculante (SI - Lactobacillus plantarum e L paracasei ssp. paracasei. Estudaram-se: as frações solúvel (A, potencialmente degradável (B e não degradável (C; a taxa de degradação da fração B (c; a degradabilidade efetiva (DE e potencial (DP, para os tratamentos: T1 (SMSE+20% CA -SI; T2 (SMSE+20% CA -CI; T3 (SMSE+35% CASI; T4 (SMSE+35% CA-CI; T5 (SMSE+20% BSM-SI; T6 (SMSE+20% BSM-CI; T7 (SMSE+35% BSM-SI; T8 (SMSE+35% BSM-CI; T9 (SMCE-SI e T10 (SMCE-CI. As médias foram comparadas num delineamento inteiramente casualisado. Houve diferença (P0,05. A adição de BSM (20 e 35% às SMSE aumentou (PThe goal of this work was to evaluate the ruminal degradation of corn silages without spikes (CSWS, added with sugar cane (SC and dry manioc pulp (DMP, as compared as corn silage (CS. The foods were ensiled with (WI and without (IF inoculant’s, composed by Lactobacillus plantarum and L. paracasei ssp. paracasei. The following fractions have been studied: water soluble (A, potentially degradable (B, non degradable (C, fraction B degradable ratio (c; effective (ED and potential (PD degradability. The treatments evaluated were: T1 (CSWS + 20% SC -IF; T2 (CSWS + 20% SC-WI; T3 (CSWS + 35% SC -IF; T4 (CSWS + 35% SC -WI; T5 (CSWS + 20% DMP-IF; T6 (CSWS + 20% DMP -WI; T7 (CSWS + 35% DMP -IF; T8 (CSWS + 35% DMP-WI; T9 (CS-WI and T10 (CS-IF. The average differences have been compared on a completely randomized model. Difference (P 0.05 for all treatments. Dry manioc pulp added to the non spikes corn silage (20 and 35% - DM basis, has allowed the rise of the DM fraction B, indicating the possibility to add up to 35% of dry manioc pulp on corn silage, inst ead of green spikes.

  5. Ruminal degradability In Situ of nutritive components of some / Degradabilidade ruminal In Situ dos componentes nutritivos de alguns suplementos concentrados usados na alimentação de bovinos

    Directory of Open Access Journals (Sweden)

    Filipe Alexandre Boscaro de Castro

    2009-07-01

    Full Text Available Degradability “in situ” of dry matter (DM, organic matter (OM, crude protein (CP, neutral detergent fiber (NDF and acid detergent fiber (ADF of whole cotton seed (WCS, soybean meal (SM, sunflower seed (SS, turnip cake (TC and ground corn (GC were determined with incubation of nylon bags in the rumen of five steers ½ Simmental-Zebu. The effective degradability (ED of dry matter and crude protein with 5%/h of passage rate were respectively 40,93 and 48,95% for cotton seed; 67,35 and 53,20% for soybean meal; 63,38 and 72,50% for sunflower seed; 66,59 and 57,82% for turnip cake and 3,65 e 54,18% for ground corn. Through the fermentation rates of the different nutritious components determined on this experiment it was concluded that turnip cake has a potential to substitute soybean meal as a protein source in ruminant feeding; sunflower seed presents a high soluble fraction of crude protein and the cotton seed ether extract may affect negatively the degradation of dry matter and organic matter of this feed, specially when provided ground to the animals.As degradabilidades in situ da matéria seca (MS, matéria orgânica (MO, proteína bruta (PB, fibra em detergente ácido (FDA e da fibra em detergente neutro (FDN do caroço de algodão integral (CAI, farelo de soja (FS, grão de girassol (GG, torta de nabo forrageiro (TN e do milho triturado (MT foram determinadas com a incubação de sacos de náilon no rúmen de cinco bovinos ½ sangue Simental-Zebu. As degradabilidades efetivas (DE da matéria seca (MS e da proteína bruta (PB a taxa de passagem de 5%/h, foram, respectivamente, 40,93 e 48,95% para o caroço de algodão integral; 67,35 e 53,20% para o farelo de soja; 63,38 e 72,50% para o grão de girassol; 66,59 e 57,82 para torta de nabo forrageiro e 63,65 e 54,18% para o milho triturado. Por meio das taxas de fermentação dos diferentes componentes nutritivos determinadas neste experimento, pode-se concluir que a torta de nabo forrageiro

  6. Small heat shock proteins, protein degradation and protein aggregation diseases

    NARCIS (Netherlands)

    Vos, Michel J.; Zijlstra, Marianne P.; Carra, Serena; Sibon, Ody C. M.; Kampinga, Harm H.

    Small heat shock proteins have been characterized in vitro as ATP-independent molecular chaperones that can prevent aggregation of un- or misfolded proteins and assist in their refolding with the help of ATP-dependent chaperone machines (e. g., the Hsp70 proteins). Comparison of the functionality of

  7. Strategies for optimizing nitrogen use by ruminants

    DEFF Research Database (Denmark)

    Calsamiglia, S; Ferret, A; Reynolds, C K

    2010-01-01

    performance. However, optimal production and N utilization may be achieved through the understanding of the key mechanisms involved in the control of N metabolism. Key factors in the rumen include the efficiency of N capture in the rumen (grams of bacterial N per grams of rumen available N......) and the modification of protein degradation. Traditionally, protein degradation has been modulated by modifying the feed (physical and chemical treatments). Modifying the rumen microflora involved in peptide degradation and amino acid deamination offers an alternative approach that needs to be addressed. Current....... A better understanding of these key processes should allow the development of strategies to improve the efficiency of N utilization in ruminants...

  8. Improvement of Rice Straw for Ruminant Feed Through Unconventional Alkali Treatment and Supplementation of Various Protein Sources

    Directory of Open Access Journals (Sweden)

    SNO Suwandyastuti

    2010-05-01

    Full Text Available Various chemical treatments were conducted to increase the utilization of rice straw as feed for ruminant animals. Various sources of protein, minerals and energy should be added to improve the nutritive value of feeds. Two experiments were conducted in this study. The objective of the first experiments was to study the effect of chemical treatment on the ruminal fermentation products in cattle. Unconventional alkali treatment made from filtrate of a 10% rice hulls ash solution enriched with urea and minerals (treatment 1 increased volatile fatty acid (VFA production, ammonia nitrogen (NH3-N and rumen microbial protein synthesis (MPS. The maximum values of NH3-N production and rumen microbial protein synthesis were reached at 4 hours after incubation, while VFA was reached at 6 hours. The second experiment was conducted to study the increase of nutritive value of rice straw previously treated in experiment 1 through supplementation with various protein sources. Protein sources from the residues of vegetative oil production such as coconut, peanut and soybean showed higher responses compared to soy-sauce making residue and tofu making residue. The protein effluent production was highest (2.19 g/d at a VFA/NH3-N ration of 37.74 (r = 0.912. It can be recommended that protein sources from agro-industrial wastes can be used to increase the nutritive value and utilization of rice straw as ruminant feed. (Animal Production 12(2: 82-85 (2010Key Words: rice straw, rumen, fermentation

  9. Efeitos da fonte de enxofre sobre a população de protozoários e degradabilidade no rúmen Effects of sulfur source on protozoa population and ruminal degradability

    Directory of Open Access Journals (Sweden)

    A. Saran Netto

    2009-08-01

    Full Text Available Estudaram-se os efeitos de fontes de enxofre na dieta sobre os parâmetros ruminais de bovinos Nelore, utilizando-se oito novilhos canulados no rúmen e submetidos a quatro tratamentos, segundo a fonte de enxofre. Os animais do grupo-controle não receberam suplementação de enxofre. Os dos outros três grupos receberam flor de enxofre ou metionina ou carboquelatado, como suplementação. Os animais foram arraçoados com dieta total, utilizando cana-de-açúcar picada como volumoso. O período experimental foi de 76 dias, dividido em quatro subperíodos de 19 dias, dos quais 14 eram para adaptação à dieta e cinco para as colheitas. Não foram encontradas diferenças entre as fontes de enxofre na degradabilidade da matéria seca, proteína bruta e fibras em detergente ácido e neutro e no pH ruminal. O tratamento com carboquelatado resultou em maior número de protozoários ciliados e o tratamento metionina em menor contagem. O carboquelatado pode ser uma boa alternativa para a suplementação de enxofre para bovinos em confinamento.This research was carried out at FZEA/USP to compare the effects of different sulphur sources in diets of Nellore cattle on ruminal parameters. Eight steers were rumen cannulated and submitted to four treatments, using the following sulphur sources: control (no supplement, elemental sulphur, methionine, and carboquelated. Animals were fed a total ration using sugarcane as the roughage. The proportion roughage:concentrate was 40:60. The experimental period lasted 76 days, divided in four 19-days sub-periods. The first 14 days of each sub-period were used to adapt the animals to the diet and the last five days to collect the samples. During the sample period, ruminal liquid was sampled to protozoa count and pH determination. Also, nylon bags were incubated in rumen to determinate the degradability of dietary dry matter, crude protein, and acid and neutral detergent fiber. Treatments did not affect dietary dry matter

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

    Directory of Open Access Journals (Sweden)

    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.

  11. The requirements for rumen-degradable protein per unit of fermentable organic matter differ between fibrous feed sources

    Directory of Open Access Journals (Sweden)

    Michael eKreuzer

    2015-07-01

    Full Text Available Ruminant feed evaluation systems use constant minimum requirements of rumen-degradable protein (RDP and often relate this to apparently degradable organic matter (OM. However, studies with tropical forages indicate that RDP:apparently degraded OM might not be constant across high-fiber diets. This was tested with semi-continuous ruminal cultures (Rusitec using dried contrasting low-protein fiber sources: brachiaria hay (high in fiber, medium lignified, apple pomace (medium in fiber, highly lignified and sugar beet pulp (medium in fiber and lignification. Each feed was incubated at 14 g dry matter day-1 with 0, 0.85, 1.7, 3.4, 6.8, 13.6 or 27.2 mg g-1 urea. The amount of urea needed to reach a similar basal concentration of ammonia in the incubation fluid was tested with each feed in advance. Apparent fiber and OM degradability were determined after 48 h of incubation. Data was evaluated by regressions and analysis of variance. The response curve of incubation fluid ammonia to urea supplementation was similar in slope in all feeds. Plateaus in apparent OM degradability in relation to ammonia concentration were determined. The ammonia concentration where apparent OM and fiber degradability reached 95% of maximum was approached in the order of pomace < pulp < hay. With regard to fiber degradability, a plateau was reached at ≥80 g kg-1 crude protein only with hay and pomace, whilst a linear relationship existed between RDP and OM degradation for pulp. In hay the ratio RDP:OM degraded was equal to 1.6 but was only 1.0 in the other feeds. There was no obvious lack of branched short-chain fatty acids at low RDP. Thus, the hypothesis was confirmed but the demand for RDP seems even higher in tropical forage compared to food industrial byproducts. The efficiency of urea to promote apparent OM and fiber degradation was also variable. Thus, it seems that minimum thresholds of either RDP or ruminal ammonia concentration may not be reflected appropriately

  12. Aminopeptidases do not directly degrade tau protein

    Directory of Open Access Journals (Sweden)

    Hersh Louis B

    2010-11-01

    Full Text Available Abstract Background Tau hyperphosphorylation and aggregation to form intracellular neurofibrillar tangles is prevalent in a number of tauopathies. Thus there is current interest in the mechanisms involved in Tau clearance. It was recently reported that Tau can be degraded by an aminopeptidase known as the puromycin sensitive aminopeptidase (PSA. Until now PSA has been reported to only cleave peptides, with the largest reported substrates having 30-50 amino acids. We have studied this unique PSA cleavage reaction using a number of different PSA preparations. Results An N-terminally His tagged-PSA was expressed and purified from Sf9 insect cells. Although this PSA preparation cleaved Tau, product analysis with N and C terminal Tau antibodies coupled with mass spectrometry showed an endoproteolytic cleavage atypical for an aminopeptidase. Furthermore, the reaction was not blocked by the general aminopeptidase inhibitor bestatin or the specific PSA inhibitor puromycin. In order to test whether Tau hydrolysis might be caused by a protease contaminant the enzyme was expressed in E. coli as glutathione S-transferase and maltose binding protein fusion proteins or in Sf9 cells as a C-terminally His-tagged protein. After purification to near homogeneity none of these other recombinant forms of PSA cleaved Tau. Further, Tau-cleaving activity and aminopeptidase activities derived from the Sf9 cell expression system were separable by molecular sieve chromatography. When tested in a cellular context we again failed to see a PSA dependent cleavage of Tau. A commercial preparation of a related aminopeptidase, aminopeptidase N, also exhibited Tau cleaving activity, but this activity could also be separated from aminopeptidase activity. Conclusion It is concluded that PSA does not directly cleave Tau.

  13. The effects of alfalfa particle size and acid treated protein on ruminal ...

    African Journals Online (AJOL)

    This study was conducted to investigate the effects of alfalfa particle size (long vs. fine) and canola meal treated with hydrochloric acid solution (untreated vs treated) on ruminal chemical composition, liquid, particulate, escapable and non escapable phases in Zel sheep. Four ruminally cannulated sheep received a mixed ...

  14. Seminal-type ribonuclease genes in ruminants, sequence conservation without protein expression?

    NARCIS (Netherlands)

    Kleineidam, RG; Jekel, PA; Beintema, JJ; Situmorang, P

    1999-01-01

    Bovine seminal ribonuclease (BS-RNase) is an interesting enzyme both for functional and structural reasons. The enzyme is the product of a gene duplication that occurred in an ancestral ruminant. It is possible to demonstrate the presence of seminal-type genes in all other investigated ruminant

  15. Co-ensiling temperate grasses to improve protein use efficiency in ruminants

    Science.gov (United States)

    Preserving high-quality forage in cool humid regions of agricultural production remains a challenge due to potentially high levels of protein degradation during ensiling. Some forages such as red clover produce high levels of polyphenol oxidase (PPO) and o-diphenols to effectively inhibit proteolysi...

  16. Roles of protein ubiquitination and degradation kinetics in biological oscillations.

    Directory of Open Access Journals (Sweden)

    Lida Xu

    Full Text Available Protein ubiquitination and degradation play important roles in many biological functions and are associated with many human diseases. It is well known that for biochemical oscillations to occur, proper degradation rates of the participating proteins are needed. In most mathematical models of biochemical reactions, linear degradation kinetics has been used. However, the degradation kinetics in real systems may be nonlinear, and how nonlinear degradation kinetics affects biological oscillations are not well understood. In this study, we first develop a biochemical reaction model of protein ubiquitination and degradation and calculate the degradation rate against the concentration of the free substrate. We show that the protein degradation kinetics mainly follows the Michaelis-Menten formulation with a time delay caused by ubiquitination and deubiquitination. We then study analytically how the Michaelis-Menten degradation kinetics affects the instabilities that lead to oscillations using three generic oscillation models: 1 a positive feedback mediated oscillator; 2 a positive-plus-negative feedback mediated oscillator; and 3 a negative feedback mediated oscillator. In all three cases, nonlinear degradation kinetics promotes oscillations, especially for the negative feedback mediated oscillator, resulting in much larger oscillation amplitudes and slower frequencies than those observed with linear kinetics. However, the time delay due to protein ubiquitination and deubiquitination generally suppresses oscillations, reducing the amplitude and increasing the frequency of the oscillations. These theoretical analyses provide mechanistic insights into the effects of specific proteins in the ubiquitination-proteasome system on biological oscillations.

  17. A mathematical model of protein degradation by the proteasome

    NARCIS (Netherlands)

    Luciani, F.; Kesmir, C.; Mishto, M.; Or-Guil, 'M.; Boer, R.J. de

    2005-01-01

    The proteasome is the major protease for intracellular protein degradation. The influx rate of protein substrates and the exit rate of the fragments/products are regulated by the size of the axial channels. Opening the channels is known to increase the overall degradation rate and to change the

  18. Impaired protein degradation in FTLD and related disorders.

    Science.gov (United States)

    Götzl, Julia K; Lang, Christina M; Haass, Christian; Capell, Anja

    2016-12-01

    Impaired protein degradation has been discussed as a cause or consequence of various neurodegenerative diseases, such as Alzheimer's, Parkinson's and Huntington's disease. More recently, evidence accumulated that dysfunctional protein degradation may play a role in frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). Since in almost all neurodegenerative diseases, protein aggregates are disease-defining hallmarks, it is most likely that impaired protein degradation contributes to disease onset and progression. In the majority of FTD cases, the pathological protein aggregates contain either microtubuleassociated protein tau or TAR DNA-binding protein (TDP)-43. Aggregates are also positive for ubiquitin and p62/sequestosome 1 (SQSTM1) indicating that these aggregates are targeted for degradation. FTD-linked mutations in genes encoding three autophagy adaptor proteins, p62/SQSTM1, ubiquilin 2 and optineurin, indicate that impaired autophagy might cause FTD. Furthermore, the strongest evidence for lysosomal impairment in FTD is provided by the progranulin (GRN) gene, which is linked to FTD and neuronal ceroid lipofuscinosis. In this review, we summarize the observations that have been made during the last years linking the accumulation of disease-associated proteins in FTD to impaired protein degradation pathways. In addition, we take resent findings for nucleocytoplasmic transport defects of TDP-43, as discussed for hexanucleotide repeat expansions in C9orf72 into account and provide a hypothesis how the interplay of altered nuclear transport and protein degradation leads to the accumulation of protein deposits. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Ruminally undegradable protein content and digestibility for forages using the mobile bag in situ technique.

    Science.gov (United States)

    Buckner, C D; Klopfenstein, T J; Rolfe, K M; Griffin, W A; Lamothe, M J; Watson, A K; MacDonald, J C; Schacht, W H; Schroeder, P

    2013-06-01

    Four experiments were conducted to evaluate RUP content and digestibility for smooth bromegrass, subirrigated meadow, upland native range, and warm-season grasses. Samples were collected from esophageally cannulated cows or ruminally cannulated steers. Forages were ruminally incubated in in situ bags for durations of time based on 75% of total mean retention time, which was based on IVDMD and rate of passage calculations. One-half of the bags were duodenally incubated and excreted in the feces, and NDIN was analyzed on all bags for RUP calculations. Crude protein was numerically greater early in the growing cycle for grasses compared with later as grasses matured (P ≤ 0.32). The RUP was 13.3%, 13.3%, and 19.7% of CP for smooth bromegrass, subirrigated meadow, and upland native range, respectively. These values tended to be lower early in the growth cycle and increased (linear P ≤ 0.13) as forages matured for warm-season grasses and subirrigated meadows. Because both CP and RUP content change throughout the growing season, expressing RUP as a percentage of DM gives more consistent averages compared with RUP as a percentage of CP. Coefficient of variation values for RUP as a percentage of DM averaged 0.21 over all 4 experiments compared with 0.26 for RUP as a percentage of CP. Average RUP as a percentage of DM was 2.03%, 1.53%, and 1.94% for smooth bromegrass, subirrigated meadow, and upland native range, respectively. Total tract indigestible protein (TTIDP) linearly increased with maturity for subirrigated meadow samples (P Digestibility of RUP varied considerably, ranging from 25% to 60%. Subirrigated meadow, native range, and smooth bromegrass samples tended to have linear decreases (P ≤ 0.11) in RUP digestibility throughout the growing season. The amount of digested RUP was fairly consistent across experiments and averages for smooth bromegrass, subirrigated meadow, and upland native range were 0.92%, 0.64%, and 0.49% of DM, respectively. Warm

  20. Ruminal fermentation modification of protein and carbohydrate by means of roasted and estimation of microbial protein synthesis Modificação da fermentação ruminal de proteína e carboidrato por meio da tostagem e estimativa de síntese de proteína microbiana

    Directory of Open Access Journals (Sweden)

    Héctor Andrade-Montemayor

    2009-07-01

    Full Text Available The animal meal prohibition as a protein source with low ruminal degradability in ruminant nutrition, creates the need to seek alternatives, as legume seeds, however, its protein, have a high degradability, which could generate losses of nitrogen in the rumen. Other problem in the legume seeds is the content of antinutritional factors such as protease inhibitors, tannins, phenolic compounds, lectins and some others, could affect the digestibility. One alternative to decrease the degradability of the protein and/or decrease the activity of some antinutritional factors is the use of different technological treatments such as roasting. Microbial protein synthesis is dependent on the availability of nitrogen or amino acids and of the degradable carbohydrate content, or of content of Organic Matter digestible and rumen degraded (OMDRD, and the synchronization in rumen degradation between protein and carbohydrates. If there are high amounts of degradable nitrogen or if the content and rate of carbohydrates degradation in the rumen is reduced or not synchronized with the degradation of protein, could be losses of nitrogen and/or energy in the rumen. In this paper, we analyze the characteristics of proteins and carbohydrates from various seeds and grains, the effect of dry-roasted application on degradability behavior of seeds.A proibição das farinhas de origem animal como fonte de proteína com baixa degradabilidade na nutrição de ruminantes, criou a necessidade de buscar alternativas, como sementes de leguminosas, porém, sua proteínas têm alta degradabilidade, podendo gerar perdas de nitrogênio no rúmen. Outro problema em sementes de leguminosas é o conteúdo de fatores antinutricionais, como os inibidores de proteases, taninos, compostos fenólicos, lectinas e alguns outros, podendo afetar a digestibilidade. Uma alternativa para diminuir a degradabilidade da proteína e/ou diminuir a atividade de alguns fatores antinutricionais é o uso de

  1. Ruminal fermentation response and nitrogen retention from sheep fed rumen undegradable protein

    Directory of Open Access Journals (Sweden)

    Wisri Puastuti

    2012-03-01

    Full Text Available The utilization of rumen undegradable protein, could increase supply of amino acids in small intestine to meet host protein requirement. To increase the utilization of feed protein, feed protein source which is highly degradable should be protected from degradation in the rumen. The objective of the study was to increase nitrogen retention through substitution of fish meal protein with soy bean meal protected by banana stem juice observed from rumen fermentation activities. The study used 18 late pregnant Sumatera Composite breed ewes. Rations consisted of fresh chopped elephant grass, supplemented with a commercial concentrate and mineral block in the form of Comin plus and different types of protein supplement as treatment diets. The type of protein supplements were: soya bean meal (RK, soy bean meal protected with banana stem juice (RKT and fish meal (RTI. Treatment diets were offered during late pregnancy (from two months before to two months after partus, two weeks adaptation period was carried out before data recording. Design of the study used randomized complete block design. Results of the study show that rumen ammonia concentration in sheep fed on RKT was not significantly different either from RTI or from RK, however, the nitrogen retention was significantly (P 0.05, however, proportion of C2 and nC4 was significantly different between diet treatment. Methane emission was higher (70.3 mM or 37.2% higher from total energy VFA in diet RKT which is indicated that fermentation system was not efficient. It can be concluded the higher rumen ammonia concentration and lower nitrogen retention in protected soy bean meal supplement indicated that soy bean meal protected by banana stem juice in the ration was not able to substitute fish meal protein which is resistance from degradation in the rumen.

  2. The effects of starch and rapidly degradable dry matter from concentrate on ruminal digestion in dairy cows fed corn silage-based diets with fixed forage proportion.

    Science.gov (United States)

    Lechartier, C; Peyraud, J-L

    2011-05-01

    This study investigated the effects of the type (starch vs. nonstarch) and rate of ruminal degradation of carbohydrates from the concentrate on digestion in dairy cows fed corn silage-based diets. Six ruminally cannulated cows were assigned to 6 treatments in a 6 × 6 Latin square design. Treatments were arranged in a 3 × 2 factorial design. Two starch levels [25 and 41% dry matter (DM) for low starch (LS) and high starch (HS) diets, respectively] were obtained by replacing starch-rich feedstuffs by nonstarch feedstuffs. These starch levels were combined with slowly, moderately, and rapidly rumen-degradable feedstuffs to obtain 3 levels of rapidly degradable carbohydrates from concentrate (18, 23, and 28% DM). These levels were estimated from the DM disappearance of concentrate after 4h of in sacco incubation (CRDM). Wheat and corn grain were used as rapidly degradable and slowly degradable starch feedstuffs, respectively. Soybean hulls and citrus pulp were used as slowly degradable and rapidly degradable nonstarch feedstuffs, respectively. No interaction effect was found between dietary starch content and CRDM on pH range, volatile fatty acid (VFA) range, or VFA profile. Increasing CRDM led to a linear decrease in acetate-to-propionate ratio (from 2.7 to 2.1), and a linear increase in the pH and VFA ranges (from 0.86 to 1.12 pH units and from 34 to 56mM, respectively). Feeding HS diets decreased acetate-to-propionate ratio (2.6 vs. 2.0) and increased pH range (0.89 vs. 1.04 pH units), but had no effect on VFA range. Increasing CRDM linearly decreased mean ruminal pH in LS diets but linearly increased mean ruminal pH in HS diets. Fibrolytic activity was unaffected in LS diets but decreased strongly in HS diets (from 62 to 50%). These findings suggest that pH regulation differs on a short-term and on a longer-term basis. In the short-term, increasing CRDM increased the rate of VFA production, which may have been partly buffered under LS diets due to the higher

  3. Protein Degradation Rate in Arabidopsis thaliana Leaf Growth and Development.

    Science.gov (United States)

    Li, Lei; Nelson, Clark J; Trösch, Josua; Castleden, Ian; Huang, Shaobai; Millar, A Harvey

    2017-02-01

    We applied 15N labeling approaches to leaves of the Arabidopsis thaliana rosette to characterize their protein degradation rate and understand its determinants. The progressive labeling of new peptides with 15N and measuring the decrease in the abundance of >60,000 existing peptides over time allowed us to define the degradation rate of 1228 proteins in vivo. We show that Arabidopsis protein half-lives vary from several hours to several months based on the exponential constant of the decay rate for each protein. This rate was calculated from the relative isotope abundance of each peptide and the fold change in protein abundance during growth. Protein complex membership and specific protein domains were found to be strong predictors of degradation rate, while N-end amino acid, hydrophobicity, or aggregation propensity of proteins were not. We discovered rapidly degrading subunits in a variety of protein complexes in plastids and identified the set of plant proteins whose degradation rate changed in different leaves of the rosette and correlated with leaf growth rate. From this information, we have calculated the protein turnover energy costs in different leaves and their key determinants within the proteome. © 2017 American Society of Plant Biologists. All rights reserved.

  4. Effects of Synchronization of Carbohydrate and Protein Supply in Total Mixed Ration with Korean Rice Wine Residue on Ruminal Fermentation, Nitrogen Metabolism and Microbial Protein Synthesis in Holstein Steers

    Directory of Open Access Journals (Sweden)

    Min Yu Piao

    2012-11-01

    Full Text Available Three Holstein steers in the growing phase, each with a ruminal cannula, were used to test the hypothesis that the synchronization of the hourly rate of carbohydrate and nitrogen (N released in the rumen would increase the amount of retained nitrogen for growth and thus improve the efficiency of microbial protein synthesis (EMPS. In Experiment 1, in situ degradability coefficients of carbohydrate and N in feeds including Korean rice wine residue (RWR were determined. In Experiment 2, three total mixed ration (TMR diets having different rates of carbohydrate and N release in the rumen were formulated using the in situ degradability of the feeds. All diets were made to contain similar contents of crude protein (CP and neutral detergent fiber (NDF but varied in their hourly pattern of nutrient release. The synchrony index of the three TMRs was 0.51 (LS, 0.77 (MS and 0.95 (HS, respectively. The diets were fed at a restricted level (2% of the animal’s body weight in a 3×3 Latin-square design. Synchronizing the hourly supply of energy and N in the rumen did not significantly alter the digestibility of dry matter, organic matter, crude protein, NDF or acid detergent fiber (ADF (p>0.05. The ruminal NH3-N content of the LS group at three hours after feeding was significantly higher (p0.05. In addition, the purine derivative (PD excretion in urine and microbial-N production (MN among the three groups were not significantly different (p>0.05. In conclusion, synchronizing dietary energy and N supply to the rumen did not have a major effect on nutrient digestion or microbial protein synthesis (MPS in Holstein steers.

  5. Leucaena leucocephala IN RUMINANT NUTRITION

    Directory of Open Access Journals (Sweden)

    M. Barros-Rodríguez

    2014-08-01

    Full Text Available It is a common situation in extensive ruminant production systems in tropical countries to have low production indicators due to nutrient deficiencies in the diet. An economic alternative to increase animal production is the incorporation of legumes (fodder and fruits in the diet. This review, presents an analysis of the positive and negative effects of Leucaena leucocephala consumption by ruminants, with particular emphasis on the secondary compound mimosine. Leucaena due to its high nutrient content, rumen by-pass protein supply and its possible effect on the reduction of greenhouse gas (attributed to tannins has become one of the legumes most commonly used in ruminant feeding practices. However, in countries where leucaena has been introduced, its use is still limited to levels below 30% inclusion in the diet, due to the secondary compound mimosine and its isomers (3,4 and 2,3 DHP, which can induce toxicity, even when animals are inoculated with rumen fluid containing the bacteria Synergistes jonesii reported as responsible for degrading these compounds in the rumen. In the Yucatan Peninsula, Mexico, ruminants consuming leucaena can tolerate more than 50% inclusion in the diet, without having a negative impact on production, attributed intake to mimosine and its isomers. We conclude that in animals not adapted, the intake would be limited to low inclusion levels (less than 30% inclusion in the diet, mainly because of mimosine and its derivatives. The decrease in intake or diet digestibility seem to better explain the reduction in methane production, however, in vivo studies are required to clearly establish the mechanism of action. It has been reported the presence of different bacteria to S. jonessi that would have the ability to degrade mimosine and its derivatives, however, the activity of these bacteria and its effectiveness must be confirmed in vivo.

  6. Avaliação de Dois Métodos in vitro para Determinar a Cinética Ruminal e a Digestibilidade Intestinal da Proteína de Vários Alimentos Application of Two in Vitro Methods for Evaluation of Ruminal Disappearance and Intestinal Digestibility of Feed Protein

    Directory of Open Access Journals (Sweden)

    Fernando Iván Londoño Hernández

    2002-02-01

    undegradable protein was not constant. The inhibitor method was more efficient to evaluate and predict kinetic parameters of ruminal degradation and the three-step method for intestinal digestibility of ruminal undegradable protein.

  7. Degradabilidade ruminal da fibra das frações do resíduo industrial de tomate Ruminal degradability of fiber of tomato industrial residues fractions

    Directory of Open Access Journals (Sweden)

    W.E. Campos

    2007-02-01

    Full Text Available Avaliou-se a degradabilidade in situ da fibra em detergente neutro (FDN e da fibra em detergente ácido (FDA das cascas e das sementes inteiras ou moídas (peneira de 2mm do resíduo industrial de tomate (RIT. Foram utilizados três bovinos adultos, machos, em delineamento de blocos ao acaso com parcelas subdivididas. A solubilidade da FDN apresentou baixos valores para todas as frações estudadas, sendo semelhante entre cascas (3%, sementes inteiras (5% e resíduo (7%. Sementes moídas apresentaram maior solubilidade da FDN (20%. Os potenciais de degradação da FDN para sementes moídas, RIT, cascas e sementes inteiras foram de 78, 64, 36 e 33%, respectivamente. As taxas de degradação da FDN variaram de 4,4 a 14,8%/h para as diferentes frações do RIT, sendo de 9%/h para o resíduo integral. Os parâmetros de degradação da FDA seguiram a mesma tendência da FDN. Concluiu-se que, apesar de a FDN e a FDA do RIT terem apresentado altas taxas de degradação, o potencial de degradação dessas frações depende do processamento, pois sementes inteiras apresentaram degradabilidades muito inferiores às das sementes moídas.The in situ procedure was used to evaluate the disappearance of neutral detergent fiber (NDF and acid detergent fiber (ADF of skins and whole or grounded (sieve of 2mm seeds of tomato by-product (TBP. Three steers were used in a split-plot block design. The solubility of NDF showed low values for all the evaluated fractions, been similar among skins (3%, whole seeds (5% and total TBP (7%. Grounded seeds showed higher NDF solubility (20%. The NDF potential degradability for grounded seeds, TBP, skins and whole seed were 78, 64, 63 and 33%, respectively. NDF degradability rates varied from 4.4 to 14.8%/h for all TBP fractions, been 9%/h for total TBP. The ADF parameters followed the same trend as NDF. TBP NDF and ADF showed good degradability rates, but the potential degradability of these fractions depends on the showed

  8. 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. L.J. Erasmus· and J.Prinsloo. Animal and Dairy Science Research Institute, Private Bag X2, Irene, 1675Republic of South Africa ..... amino acid profile when compared to most vegetable protein sources ...

  9. Sequence-based analysis of protein degradation rates

    NARCIS (Netherlands)

    Correa Marrero, Miguel; Dijk, van Aalt-Jan; Ridder, de Dick

    2017-01-01

    Protein turnover is a key aspect of cellular homeostasis and proteome dynamics. However, there is little consensus on which properties of a protein determine its lifetime in the cell. In this work, we exploit two reliable datasets of experimental protein degradation rates to learn models and uncover

  10. Níveis de proteína na dieta de bovinos Nelore de três condições sexuais: consumo, digestibilidades total e parcial, produção microbiana e parâmetros ruminais Increasing crude protein levels in diets of Nellore cattle: intake, ruminal and total tract digestibility, microbial protein synthesis, and ruminal metabolism

    Directory of Open Access Journals (Sweden)

    Robson Magno Liberal Véras

    2007-08-01

    .Twelve Nellore cattle fitted with ruminal and duodenal cannulas were used to investigate the effects of increasing dietary crude protein level (7, 10, 13, and 15% on intake, ruminal and total tract digestibility of nutrients, microbial protein synthesis, degradabilities of corn silage, corn starch, and cottonseed meal, and ruminal metabolism. Animals were blocked by sexual category (heifers, bulls and steers averaging 254.8, 285.1, and 265.6 kg of body weight, respectively and then assigned to one out of three Latin squares. Samples were incubated in the rumen of one animal of each sexual category to estimate ingredient degradability after determination of DM, CP and NDF in the incubation residues. Ruminal fluid was collected before and after-feeding to determine pH and concentration of ammonia while ruminal samples for bacterial isolation were taken 6 hours post-feeding. Blood (BUN and urine (UU were also collected for measurement of urea concentration. The degradation rates of DM and CP were both lower in corn silage than on corn starch and cottonseed meal. The interaction sexual condition x dietary crude protein level was not significant for nutrient intake. With the exception of EE and NFC, no other significant effect of dietary CP level was observed for daily intake of nutrients. Increasing dietary CP level affected total tract digestibilities of DM, OM, CP, NDF, and NFC and also resulted in higher concentrations of BUN and UU. A significant dietary crude protein level x collection time interaction was observed for ruminal NH3 but not for ruminal pH. It was concluded that increasing dietary CP levels affected intake and total tract digestibility of most nutrients.

  11. The effect of dietary protein degradability and rumen inert fat ...

    African Journals Online (AJOL)

    Unknown

    sasas.co.za/Sajas.html. 30. The effect of dietary protein degradability and rumen inert fat supplementation on calf performance ... mass gain, feed intake and feed conversion ratios in veal calves, as well as digestibility coefficients, carcass masses.

  12. Evaluation of rice straw fermented by trichoderma viride, rhizopus stolonifer and pycnoporus sanguineus by in situ ruminal degradability

    OpenAIRE

    Endres, Eliane; Thomas, Robert Wayne Steven Philip; Prates, Enio Rosa

    1999-01-01

    O melhoramento nutricional da palha de arroz fermentada por Trichoderma viride, Rhizopus stolonifer e Pycnoporus sanguineus foi avaliado por intermédio da degradação ruminal in situ. A fermentação da palha com T. viride e água por duas semanas aumentou a taxa de degradação ruminal em, aproximadamente, 31,5 e 264%, respectivamente, quando comparada com controle e água e com fermentação da palha por uma semana. Houve, também, redução no tempo de degradação efetiva da matéria seca da palha de ar...

  13. Degradação ruminal dos tecidos vegetais e composição bromatológica de cultivares de Axonopus scoparius (Flüegge Kuhlm. e Axonopus fissifolius (Raddi Kuhlm Ruminal tissue degradation and bromatological composition in Axonopus scoparius (Flüegge Kuhlm. and Axonopus fissifolius (Raddi Kuhlm cultivars

    Directory of Open Access Journals (Sweden)

    Lenir Maristela Silva Lima

    2001-06-01

    digestion kinetics and the Near Infra Red Spectroscopy (NIRS technique to determine the bromatological composition. Tissue degradation was histologically observed after the incubation of small stem and leaf portions in cattle ruminal liquid. With mature age, it was observed that Axonopus scoparius cultivars reduced the effective degradability, which was related to cell wall thickening and increased lignified tissue area. The level of crude protein was the parameter mostly affected as species grew older. Leaf fractions (limbo, keel and sheath of Axonopus fissifolius showed large areas of lignified tissues, resulting in lower digestibility rates as compared to Axonopus scoparius. The Axonopus scoparius "verde" cultivar, proved more susceptible to ruminal digestion than "roxo" cultivar, whereas no differences were observed in Axonopus fissifolius cultivars. The stem was the part of plant presenting greater reduction in digestion as compared to the leaf portion.

  14. Kinetics of ruminal degradation of alfalfa and Tifton-85 hays, and of corn SilageCinética de degradação ruminal dos fenos de alfafa e Tifton-85 e da silagem de milho

    Directory of Open Access Journals (Sweden)

    Geraldo Tadeu dos Santos

    2011-07-01

    Full Text Available The shunt portosystemic or portosystemic deviation (PSD are unique or multiples vascular communications between the systemic circulaton and the portal circulation, that permit blood flow reaches the circulatory system without first passing trhought the hepatic metabolization. May be acquired or congenital and can also be classified as intrahepatic located within the liver or extrahepatic located outside the liver parenquima. The acquired form is usually associated with intra-hepatic disorders. They usually suggest tortuous vessels that communicate with the caudal vena cava in the region of the left kidney. The congenital form is associated with genetic lineage and one of the most affected is Maltese breed. This case report describes the diagnostis and treatment of a eight year-old female Maltese dog presenting extrahepatic Portosystemic Shunt. The patient showed signs of hepatic encephalopathy, such as restlessnees, weakness, deambulation, head tremor and impaired visual. Complementary exam demonstrated: postprandial glicemia near the lower limit of reference, alkaline phosphatase (ALP and alanine aminotransferase (ALT increased and hipoalbuminemia. Ultrasonography revealed the presence of vesical calculus and bilateral kidney, liver decreases and increased echogenicity, gallbladder with anechoic content and high cellularity may suggest liver/ colangiohepatopatia and extrahepatic DPS was detected. The use of Doppler assisted in identifying the location of the bypass communication and the turbulence detecting the extrahepatic DPS. Protein-restricted diet and antibiotic therapy with amoxicillin achieved good results. The clinical treatment was decided make only the clinical and maintain the quality of life of the patient. Objetivou-se avaliar a degradabilidade ruminal e a taxa de degradação in situ da matéria seca (MS, da proteína bruta (PB e da fibra em detergente neutro (FDN, dos fenos de alfafa e Tifton-85 e da silagem de milho. Foram

  15. Características físicas e degradação ruminal da palha de arroz amonizada e consumo seletivo em ovinos Physical characteristics and ruminal degradation of ammoniated rice straw and selective intake by sheep

    Directory of Open Access Journals (Sweden)

    Vagner de Alencar Arnaut de Toledo

    1998-10-01

    Full Text Available O experimento foi conduzido no Setor de Digestibilidade do Departamento de Zootecnia, da Universidade Estadual de Maringá, Paraná, Brasil, com o objetivo de caracterizar o consumo seletivo por ovinos de palha de arroz amonizada, através da degradação ruminal e de algumas características físicas das sobras da palha. Os parâmetros medidos foram: volume de empacotamento (VE, ml/g de MS, retenção de água (RA, ml/g de MS e solubilidade em água da MS (SOL %. Utilizaram-se 15 ovinos, adultos, machos, castrados, com peso médio de 63,8kg. O delineamento experimental foi completamente casualizado com 3 repetições. As características físicas das sobras de MS não foram afetadas pelo nível de oferta (P > 0,05, sendo os valores médios para VE, RA e SOL de 3,22ml/g, 8,42ml/g e 12,0%, respectivamente. Os parâmetros de degradabilidade da MS também não sofreram efeitos dos níveis de oferta (P > 0,05, sendo as médias de 23,3%/h, 47,8%/h e 3,38%/h para os parâmetros a, b e c, respectivamente. A avaliação das características físicas e degradabilidade da MS das sobras não esclareceu o efeito do aumento dos níveis de oferta de palha amonizada no consumo seletivo de matéria seca por ovinos.The study was carried out at Sector of Digestibility, Department of Animal Science of Universidade Estadual de Maringá, Maringá, State of Paraná, Brazil, to instigate the selective intake of ammoniated rice straw by sheep through ruminal degradation and some physical features of leftovers. The following parameters were studied: packing volume (PV, DM ml/g, water retention (WR DM ml/g and DM solubility in water (SOL %. Fifteen male castrated adult sheep with mean body weight of 63.8kg were used as a sample, in a completely randomized design with three replications. The physical characteristics of DM leftovers were not affected by feeding levels (P > 0.05, while PV, WR and SOL mean values were 3.22ml/g, 8.42ml/g and 12.0% respectively. Neither DM

  16. Influence of yeast (Saccharomyces cerevisiae as Yea-Sacc or Levaferm) on in Sacco dry matter degradability and ruminal parameters of variously fed small ruminants.

    Science.gov (United States)

    Flachowsky, G; Tiroke, K; Matthey, M

    1992-01-01

    Two series of experiments with rumen fistulated castrated male sheep and goats were carried out. In experiment I three sheep each consumed rations rich in concentrate (700 g concentrate, 200 g chopped wheat straw) or roughage (700 g artificially dried ryegrass, 200 g chopped wheat straw per animal per day) and supplemented with 0, 1, 2 or 4 g Yea-Sacc (Saccharomyces cerevisiae; USA) per sheep per day. In experiment II three sheep were fed with 1000 g artificially dried ryegrass and 200 g concentrate, three goats consumed 750 g ryegrass and 150 g concentrate. 0, 0.5, 1 or 2 g Levaferm (Saccharomyces cerevisiae; Germany) per animal per day were added. Rations of all animals were supplemented with minerals and vitamins. After 14 days of feeding wheat straw, ammonia treated wheat straw and artificially dried grass (exp. I) or wheat straw and artificially dried grass (exp. II) were incubated in nylon bags in the rumen for 6, 12, 24, 48 and 72 hours. At the end of the experiments rumen fluid was taken via cannulae and parameters of rumen fermentation were measured. Higher levels of added Yea-Sacc decreased in sacco dry matter degradability of all incubated feeds. Depression was much higher if Yea-Sacc was added to the concentrate ration (overall mean for 24, 48 and 72 h incubation time: 55.1, 47.1, 46.1 and 44.5 for 0, 1, 2 and 4 g Yea-Sacc) than to the roughage diet (58.7, 56.3, 55.0 and 54.1%). Levaferm did not significantly influence the rumen dry matter degradability of incubated feeds (overall mean for 24, 48, and 72 h incubation time: 64.0; 64.9; 64.9 and 64.2% for sheep; 63.0; 63.2; 63.2 and 61.6% for goats, if added with 0, 0.5, 1 and 2 g Levaferm per animal per day). Rumen pH, concentration of volatile fatty acids and molar concentration of fatty acids in rumen fluid were not significantly influenced by added yeasts. More research seems necessary to find out the mode of action of yeast and to quantify and to reproduce the effects of added yeast.

  17. Ruminal degradability of oat (Avena sativum L., Coast cross (Cynodon dactilon L., and Esmeralda grass (Joysia japonica hay pellet or not/ Degradabilidade ruminal dos fenos de aveia (Avena sativum L, Coast cross (Cynodon dactilon L., e grama Esmeralda (Joysia japonica peletizado ou não

    Directory of Open Access Journals (Sweden)

    Teresa Cristina Alves

    2006-06-01

    Full Text Available The objective of this work was to evaluate the ruminal degradability of dry matter (DM, crude protein (CP, neutral detergent fiber (NDF and acid detergent fiber (ADF of the oat hay (OH, Coast cross hay (CCH, pellet Esmeralda grass hay (PEH and Esmeralda grass hay (EH. The in situ technique was used, with four bovines, castrated, rumen fistulated, distributed in an Latin square 4x4 experimental design. The material was incubated in the rumen in the times 0, 6, 24, 48, 72 and 144 hours. There was no difference between hays on the degradation rate (c, %/h for the CP, NDF and ADF. For the DM, the OH and PEH presented higher degradation rate. There was difference on the effective degradability (ED, % of the hays, and the observed values were: OH, 48.74%; PEH, 42.44%; EH, 35.13%; CCH, 30.24%. It can be concluded that oat hay presented the highest values for the soluble fraction and for the ED of all nutrients evaluated. The pellet process of the Esmeralda grass resulted in higher ED of the DM and of the fiber, with no alteration on the CP ED.O objetivo deste trabalho foi avaliar a degradabilidade ruminal da matéria seca (MS, proteína bruta (PB, fibra em detergente neutro (FDN e fibra em detergente ácido (FDA do feno de aveia (FA, feno de Coast cross (FCC, feno de grama Esmeralda peletizado (FEP e feno de grama Esmeralda (FE. Foi utilizada a técnica in situ com quatro bovinos machos, castrados, fistulados no rúmen, distribuídos em um delineamento experimental em quadrado latino 4x4. O material foi incubado no rúmen nos tempos de 0, 6, 24, 48, 72 e 144 horas. Não houve diferença entre os fenos na taxa de degradação (c, %/h para a PB, FDN e FDA. Para a MS, o FA e FEP apresentaram maior taxa de degradação. Houve diferença na degradabilidade efetiva (DE da MS dos fenos sendo os valores observados: FA, 48,74%; FEP, 42,44%; FE, 35,13%; FCC, 30,24%. Conclui-se que o feno de aveia obteve maiores valores para a fração solúvel e maior

  18. Protein molecular structures in alfalfa hay cut at three stages of maturity and in the afternoon and morning and relationship with nutrient availability in ruminants.

    Science.gov (United States)

    Yari, Mojtaba; Valizadeh, Reza; Naserian, Abbas Ali; Jonker, Arjan; Yu, Peiqiang

    2013-09-01

    Molecular structures in feed protein influence its digestive behavior, availability and utilization. From a nutritive point of view, stage of maturity and cutting time are important factors affecting nutrient profiles and availability of alfalfa (Medicago sativa L.) hay in ruminants. The objectives of this study were to determine protein molecular structures by Fourier transform infrared spectroscopy (FTIR), and their relationship with nutrient profiles and availability in ruminants of alfalfa hay cut at early bud, late bud and early flower stages and in afternoon and morning. With advancing maturity, molecular structure ratios of α-helix:β-sheets decreased, while amide I:amide II increased (P ≤ 0.05). Alfalfa cutting in afternoon versus morning increased protein structure α-helix:β-sheets and α-helix:others ratios (P alfalfa hay changed with advancing maturity and during the day and these protein structures affected predicted nutrient availability of alfalfa hay in ruminants. © 2013 Society of Chemical Industry.

  19. Effects of feeding canola meal or wheat dried distillers grains with solubles as a major protein source in low- or high-crude protein diets on ruminal fermentation, omasal flow, and production in cows.

    Science.gov (United States)

    Mutsvangwa, T; Kiran, D; Abeysekara, S

    2016-02-01

    The objective of this study was to determine the effects of feeding canola meal (CM) or wheat dried distillers grains with solubles (W-DDGS) as the major source of protein in diets varying in crude protein (CP) content on ruminal fermentation, microbial protein production, omasal nutrient flow, and production performance in lactating dairy cows. Eight lactating dairy cows were used in a replicated 4×4 Latin square design with 29-d periods (21 d of dietary adaptation and 8 d of measurements) and a 2×2 factorial arrangement of dietary treatments. Four cows in 1 Latin square were ruminally cannulated to allow ruminal and omasal sampling. The treatment factors were (1) source of supplemental protein (CM vs. W-DDGS) and (2) dietary CP content (15 vs. 17%; DM basis). Diets contained 50% forage and 50% concentrate, and were fed twice daily at 0900 and 1600 h as total mixed rations for ad libitum intake. Dry matter intake and milk yield were unaffected by dietary treatments; however, milk yield in cows that were fed CM was numerically greater (+1.1 kg/d) when compared with cows fed W-DDGS. Feeding CM increased milk lactose content compared with feeding W-DDGS. Milk urea nitrogen and ruminal NH3-N concentrations were greater in cows fed the high-CP compared with those fed the low-CP diet. The rumen-degradable protein supply was greater in cows fed the high-CP when compared with those fed the low-CP diet when diets contained CM, whereas rumen-degradable protein supply was lower in cows fed the high-CP when compared with those fed the low-CP diet when diets contained W-DDGS. Total N flow at the omasal canal was not affected by diet; however, omasal flow of NH3-N was greater in cows fed CM when compared with those fed W-DDGS. The rumen-undegradable protein supply was greater in cows fed the low-CP when compared with those fed the high-CP diet when diets contained CM, whereas rumen-undegradable protein supply was lower in cows fed the low-CP when compared with those fed the

  20. Associations among dietary non-fiber carbohydrate, ruminal microbiota and epithelium G-protein-coupled receptor, and histone deacetylase regulations in goats.

    Science.gov (United States)

    Shen, Hong; Lu, Zhongyan; Xu, Zhihui; Chen, Zhan; Shen, Zanming

    2017-09-19

    Diet-derived short-chain fatty acids (SCFAs) in the rumen have broad effects on the health and growth of ruminants. The microbe-G-protein-coupled receptor (GPR) and microbe-histone deacetylase (HDAC) axes might be the major pathway mediating these effects. Here, an integrated approach of transcriptome sequencing and 16S rRNA gene sequencing was applied to investigate the synergetic responses of rumen epithelium and rumen microbiota to the increased intake of dietary non-fiber carbohydrate (NFC) from 15 to 30% in the goat model. In addition to the analysis of the microbial composition and identification of the genes and signaling pathways related to the differentially expressed GPRs and HDACs, the combined data including the expression of HDACs and GPRs, the relative abundance of the bacteria, and the molar proportions of the individual SCFAs were used to identify the significant co-variation of the SCFAs, clades, and transcripts. The major bacterial clades promoted by the 30% NFC diet were related to lactate metabolism and cellulose degradation in the rumen. The predominant functions of the GPR and HDAC regulation network, under the 30% NFC diet, were related to the maintenance of epithelium integrity and the promotion of animal growth. In addition, the molar proportion of butyrate was inversely correlated with the expression of HDAC1, and the relative abundance of the bacteria belonging to Clostridum_IV was positively correlated with the expression of GPR1. This study revealed that the effects of rumen microbiota-derived SCFA on epithelium growth and metabolism were mediated by the GPR and HDAC regulation network. An understanding of these mechanisms and their relationships to dietary components provides better insights into the modulation of ruminal fermentation and metabolism in the promotion of livestock production.

  1. Degradação ruminal da silagem de milho e da palha de arroz utilizando enzimas fibrolíticas exógenas - DOI: 10.4025/actascianimsci.v30i4.6462 Ruminal degradation of corn silage and rice straw using exogenous fibrolityc enzymes - DOI: 10.4025/actascianimsci.v30i4.6462

    Directory of Open Access Journals (Sweden)

    Telma Teresinha Berchielli

    2009-03-01

    Full Text Available Estudou-se o efeito da adição de enzimas fibrolíticas (celulase e xilanase sobre a degradabilidade in situ da matéria seca (MS, proteína bruta (PB, fibra em detergente neutro (FDN, fibra em detergente ácido (FDA e hemicelulose da silagem de milho (SM e da palha de arroz (PA, utilizando seis bovinos fistulados no rúmen. Também foi avaliada a proporção de nitrogênio (N insolúvel em detergente ácido em relação ao N total (NIDA/N total. As enzimas fibrolíticas foram extraídas dos fungos Aspergillus Níger e Trichoderma longibrachiatum, sendo fornecido 0,75 g kg-1 de MS animal-1 dia-1, via cânula ruminal. Os tempos de incubação foram: 0, 3, 6, 12, 24, 48, 72 e 96 h. Os resíduos de incubação foram comparados por meio da microscopia eletrônica de varredura (MEV. A fração solúvel da MS e da PB da silagem de milho aumentou com a adição de enzimas fibrolíticas. Não houve efeito das enzimas sobre a degradação da parede celular dos volumosos. A adição de enzimas não alterou o teor de NIDA/N total da silagem de milho, porém aumentou esta proporção nos resíduos da palha de arroz incubados durante 12 e 48 h. As observações ao MEV indicaram aumento da colonização bacteriana sobre a parede celular dos volumosos, mas sem efeito sobre a degradação ruminal.This study evaluated the effect of fibrolytic enzyme addition (cellulase and xylanase on the dry matter (DM, crude protein (CP, neutral detergent fiber (NDF, acid detergent fiber (ADF and hemicellulose in situ degradability of corn silage (CS and rice straw (RS, using six cattle cannulated on the rumen. Acid detergent insoluble nitrogen (ADIN/total N was evaluated. The fibrolytic enzymes were extracted from Aspergillus niger and Trichoderma longibrachiatum fungi, supplied at 0.75 g/kg of DM/animal/day, through ruminal cannula. The incubation times were 0, 3, 6, 12, 24, 48, 72 and 96 hours. The incubation residues were compared by scanning electron micrograph (SEM

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

    African Journals Online (AJOL)

    low-degradable protein sources. Diet A was formulated with only natural protein sources (55% UDP),diet A1 contained 1,6% urea in the total diet (41% UDP) and diet A2 contained 1,0% urea in the total diet. (47% UDP).The experimental diets were allocated to 21 cows per treatment from week 4 through 33 of lactation.

  3. 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.

  4. Parâmetros cinéticos da degradação in vitro de alimentos incubados com inóculo microbiano de diferentes espécies de ruminantes Kinetic parameters of the ruminal in vitro degradation of feedstuffs given to different ruminant species

    Directory of Open Access Journals (Sweden)

    A.R.G.F. Bezerra

    2005-08-01

    Full Text Available Parâmetros cinéticos da degradação ruminal de alguns alimentos utilizados para ruminantes de zoológicos foram estimados mediante incubação in vitro com líquido ruminal de audade (Ammotragus lervia, cervo sambar (Cervus unicolor, elande (Taurotragus oryx, bovino (Bos taurus, bubalino (Bubalus bubalis, caprino (Capra hircus e ovino (Ovis aries. Os parâmetros cinéticos foram estimados pela técnica da produção de gás, cujos dados foram ajustados pelos modelos de um e de duplo compartimento. Não foram detectadas diferenças nos parâmetros cinéticos que permitissem agrupar os alimentos (fibrosos × não fibrosos e os animais (domésticos × silvestres. O modelo de duplo compartimento foi o mais adequado para a estimação dos parâmetros cinéticos da degradação ruminal. Inóculo microbiano oriundo de ruminantes domésticos não é recomendado para estimar parâmetros cinéticos da degradação ruminal de alimentos utilizados para ruminantes silvestres de zoológicos.The estimation of the ruminal kinetic parameters of pumpkin, potato-sweet, beet, broccoli, carrot, alfalfa hay, alfalfa pellet and bean, currently used for feeding wild and domestic ruminants raised in the Rio de Janeiro Zoo, was made through in vitro incubation of the feedstuffs together with ruminal fluid obtained from aoudad (Ammotragus lervia, sambar deer (Cervus unicolor, eland (Taurotragus oryx, cattle (Bos taurus, buffalo (Bubalus bubalis, goat (Capra hircus and sheep (Ovis aries. The gas production technique was used to obtain gas profiles, and the data were fitted by the mono or double compartmental model. The kinetic parameters were discrepant among both, animals and feedstuffs, and the double compartmental model gave the best estimation. Ruminal inocula from domestic ruminants can not be used to estimate the kinetic parameters of ruminal degradation of feedstuffs for wild ruminants.

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

    Directory of Open Access Journals (Sweden)

    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.

  6. 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 ...

  7. Degradabilidade ruminal e digestibilidade intestinal de alimentos por intermédio da técnica in situ associada à do saco de náilon móvel Ruminal degradability and intestinal digestibility of feeds by means of associated technical in situ and mobile nylon bag

    Directory of Open Access Journals (Sweden)

    Marcus Vinicius Morais de Oliveira

    2003-12-01

    Full Text Available Objetivou-se no presente trabalho verificar a degradação ruminal e a digestibilidade intestinal e total da matéria seca (MS e da proteína bruta (PB do farelo de soja, do grão de milho, do melaço em pó, da farinha de peixe, da farinha de penas e do feno de alfafa, por intermédio da técnica de degradabilidade ruminal in situ associada à técnica do saco de náilon móvel. As amostras dos alimentos foram moídas e colocadas em duplicata em sacos de náilon de 10x5 cm (48 micras nas quantidades de 15 e 5 mg de MS/cm² para os alimentos concentrados e feno de alfafa, respectivamente. Os sacos de náilon permaneceram incubados no rúmen de bois holandeses por 0; 2; 6; 8; 24 e 48 h; e 0; 8; 12; 24; 48; 72 e 96 horas, respectivamente, sendo depois retirados e sua duplicata inserida no duodeno através de uma cânula. Posteriormente, os sacos foram coletados junto com as fezes. Os valores de degradabilidade efetiva da PB para uma velocidade de passagem de 5%/hora, para o melaço em pó, grão de milho, farelo de soja, farinha de peixe, farinha de penas e feno de alfafa, foram de 100,00; 62,50; 57,90; 39,30; 34,20 e 60,90%, respectivamente; a digestibilidade intestinal de 100,00; 96,05; 99,79; 98,19; 96,07 e 94,64%, respectivamente; e a digestibilidade total de 100,00; 97,86; 99,87; 98,88; 97,35 e 98,09%, respectivamente. Verificou-se que as proteínas do melaço foram totalmente solúveis no rúmen, sendo as do milho, feno e farelo de soja bastante degradadas, além de possuírem um aproveitamento quase total no intestino. As proteínas das farinhas de peixe e de penas apresentaram baixa solubilidade ruminal e alta digestibilidade intestinal, sendo a farinha de peixe levemente mais digerida no intestino do que a farinha de penas.The objective of this work was to evaluate the ruminal degradation and intestinal and total dry matter (DM and crude protein (CP digestibility of soybean meal, corn grain, dry molasses, fish meal, feather meal and

  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. 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.

  11. 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.

  12. Degradation of pro-insulin-receptor proteins by proteasomes.

    Science.gov (United States)

    Cruz, Miguel; Velasco, Eduardo; Kumate, Jesús

    2004-01-01

    Type-2 diabetes is characterized by hyperinsulinemia, peripheral insulin resistance, and diminished tyrosine phosphorylation activity. It has been recently shown that proteasomes are implicated in the degradation of the insulin receptor substrate-1 (IRS-1) but not in that of the insulin receptor (IR). However, it is unknown whether proteasomes are involved in pro-IR degradation. We used CHO-IR and the 3T3-L1 cells treated with insulin at different concentrations and compared the proteasome activity of IRS-1, IR, and pro-IR degradation either in presence or in absence of lactacystin. A total of 100 nM of insulin allowed degradation of IRS-1 after 6 h of incubation. At 1,000 nM of insulin, pro-IR degradation began at 1 h of incubation, similar to IRS-1 degradation. Surprisingly, at a higher concentration (10 microM) of insulin, a drastic decrease of proteins was observed from the first minute of incubation. This activity was blocked by lactacystin, a specific proteasome inhibitor. According to these results, we propose that pro-IR is degraded by proteasomes.

  13. 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 simp...

  14. Mass Spectrometry-based Immunoassay for the Quantification of Banned Ruminant Processed Animal Proteins in Vegetal Feeds.

    Science.gov (United States)

    Steinhilber, Andreas E; Schmidt, Felix F; Naboulsi, Wael; Planatscher, Hannes; Niedzwiecka, Alicia; Zagon, Jutta; Braeuning, Albert; Lampen, Alfonso; Joos, Thomas O; Poetz, Oliver

    2018-02-22

    The ban of processed animal proteins (PAPs) in feed for farmed animals introduced in 2001 was one of the main EU measures to control the bovine spongiform encephalopathy (BSE) crisis. Currently, microscopy and polymerase chain reaction (PCR), are the official methods for the detection of illegal PAPs in feed. However, the progressive release of the feed ban, recently with the legalization of non-ruminant PAPs for the use in aquaculture, requires the development of alternative methods to determine the species origin and the source (legal or not). Additionally, discussions about the need for quantitative tests came up, particularly if the zero-tolerance-concept is replaced by introducing PAP thresholds. To address this issue, we developed and partially validated a multiplex mass spectrometry-based immunoassay to quantify ruminant specific peptides in vegetal cattle feed. The workflow comprises a new sample preparation procedure based on a tryptic digestion of PAPs in suspension, a subsequent immunoaffinity enrichment of the released peptides and a LC-MS/MS based analysis for peptide quantification using isotope labelled standard peptides. For the very first time, a mass spectrometry-based method is capable of detecting and quantifying illegal PAPs in animal feed over a concentration range of four orders of magnitude with a detection limit in the range of 0.1 % to 1 % (w/w).

  15. Colorimetric determination of in vitro feed protein degradation

    African Journals Online (AJOL)

    bound to soya-bean was bound to fish meal and corn gluten meal, respectively. Degradation values expressed as a percen- ... size of the diazonium chromophore. 'n Kolorimetriese metode vir die bepaling van ... technique involves treating feed proteins to form a highly coloured diazotized derivative, the colour of which is ...

  16. Estimation of protein degradation in rumen by three methods

    African Journals Online (AJOL)

    The rumen degradability of protein in diets containing maize straw, fish meal and 0, 30 and 60% maize grain was estimated in three ways: (i) from the difference between the total non-ammonia Nand microbial N entering the duodenum over a 24-hour period using 35S and DAPAas microbial markers,. (H) from the ...

  17. Colorimetric determination of in vitro feed protein degradation

    African Journals Online (AJOL)

    A colorimetric method for determining protein degradation us' ing a diazonium chromophore was compared with the dacron 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 ...

  18. Influence of carbohydrate source on ruminal fermentation characteristics, performance, and microbial protein synthesis in dairy cows.

    Science.gov (United States)

    Gozho, G N; Mutsvangwa, T

    2008-07-01

    Eight multiparous Holstein cows (676 +/- 57 kg of body weight; 121 +/- 17 d-in-milk) were used in a replicated 4 x 4 Latin square design to determine the effects of 4 sources of carbohydrate on milk yield and composition, ruminal fermentation, and microbial N flow to the duodenum. Four cows in one of the Latin squares were fitted with permanent ruminal cannulae. Diets contained (DM basis) 50% forage in combinations of alfalfa hay and barley silage, and 50% concentrate. The concentrate portion of the diets contained barley, corn, wheat, or oats grain as the primary source of carbohydrate. Intake of DM ranged from 24.0 to 26.2 kg/d, and it tended to be lower in cows fed the wheat-based diet compared with those fed the barley-based diet; consequently, milk yield tended to be lower in cows fed the wheat-based diet compared with those fed the barley-based diet. Cows fed the barley- or wheat-based diets had a lower milk fat content compared with those fed the corn-based diet. Ruminal fermentation characteristics were largely unaffected by the source of dietary carbohydrate, with similar ruminal pH and volatile fatty acid and ammonia concentrations for the first 6 h after the morning feeding. Dietary treatment did not affect total tract apparent digestibility of DM, organic matter, and neutral detergent fiber; however, total tract apparent digestibility of starch in cows fed the oats-based diet was higher compared with those fed the corn-and wheat-based diets. Nitrogen that was used for productive purposes (i.e., N secreted in milk + N apparently retained by the cow) tended to be lower in cows fed the wheat-based diet compared with cows fed the barley-, corn-, or oats-based diets. Urinary purine derivative (PD) excretion was similar in cows fed the barley-, corn-, and wheat-based diets; however, purine derivative excretion was higher in cows fed the barley-based diet compared with those fed the oats-based diet. Consequently, estimated microbial N flow to the duodenum was

  19. Identification of Differentially Expressed Proteins in Liver in Response to Subacute Ruminal Acidosis (SARA Induced by High-concentrate Diet

    Directory of Open Access Journals (Sweden)

    X. Y. Jiang

    2014-08-01

    Full Text Available The aim of this study was to evaluate protein expression patterns of liver in response to subacute ruminal acidosis (SARA induced by high-concentrate diet. Sixteen healthy mid-lactating goats were randomly divided into 2 groups and fed either a high-forage (HF diet or a high-concentrate (HC diet. The HC diet was expected to induce SARA. After ensuring the occurrence of SARA, liver samples were collected. Proteome analysis with differential in gel electrophoresis technology revealed that, 15 proteins were significantly modulated in liver in a comparison between HF and HC-fed goats. These proteins were found mainly associated with metabolism and energy transfer after identified by matrix-assisted laser desorption ionization/time of flight. The results indicated that glucose, lipid and protein catabolism could be enhanced when SARA occurred. It prompted that glucose, lipid and amine acid in the liver mainly participated in oxidation and energy supply when SARA occurred, which possibly consumed more precursors involved in milk protein and milk fat synthesis. These results suggest new candidate proteins that may contribute to a better understanding of the mechanisms that mediate liver adaptation to SARA.

  20. 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.

  1. Effects of Dietary Supplementation with Hainanmycin on Protein Degradation and Populations of Ammonia-producing Bacteria

    Directory of Open Access Journals (Sweden)

    Z. B. Wang

    2013-05-01

    Full Text Available An in vitro fermentation was conducted to determine the effects of hainanmycin on protein degradation and populations of ammonia-producing bacteria. The substrates (DM basis for in vitro fermentation consisted of alfalfa hay (31.7%, Chinese wild rye grass hay (28.3%, ground corn grain (24.5%, soybean meal (15.5% with a forage: concentrate of 60:40. Treatments were the control (no additive and hainanmycin supplemented at 0.1 (H0.1, 1 (H1, 10 (H10, and 100 mg/kg (H100 of the substrates. After 24 h of fermentation, the highest addition level of hainanmycin decreased total VFA concentration and increased the final pH. The high addition level of hainanmycin (H1, H10, and H100 reduced (p0.05. After 24 h of fermentation, H10 and H100 increased (p<0.05 concentrations of peptide nitrogen and AA nitrogen and proteinase activity, and decreased (p<0.05 NH3-N concentration and deaminase activity compared with control. Peptidase activitives were not affected by hainanmycin. Hainanmycin supplementation only inhibited the growth of Butyrivibrio fibrisolvens, which is one of the species of low deaminative activity. Hainanmycin supplementation also decreased (p<0.05 relative population sizes of hyper-ammonia-producing species, except for H0.1 on Clostridium aminophilum. It was concluded that dietary supplementation with hainanmycin could improve ruminal fermentation and modify protein degradation by changing population size of ammonia-producing bacteria in vitro; and the addition level of 10 mg/kg appeared to achieve the best results.

  2. Fermentation Characteristics, Tannin Contents and Ruminal Degradation of Green Tea and Black Tea By-products Ensiled at Different Temperatures

    Directory of Open Access Journals (Sweden)

    Makoto Kondo

    2014-07-01

    Full Text Available Green and black tea by-products, obtained from ready-made tea industry, were ensiled at 10°C, 20°C, and 30°C. Green tea by-product silage (GTS and black tea by-product silage (BTS were opened at 5, 10, 45 days after ensiling. Fermentation characteristics and nutrient composition, including tannins, were monitored and the silages on day 45 were subjected to in vitro ruminal fermentation to assess anti-nutritive effects of tannins using polyethylene glycol (PEG as a tannin-binding agent. Results showed that the GTS and BTS silages were stable and fermented slightly when ensiled at 10°C. The GTS stored at 20°C and 30°C showed rapid pH decline and high acetic acid concentration. The BTS was fermented gradually with moderate change of pH and acid concentration. Acetic acid was the main acid product of fermentation in both GTS and BTS. The contents of total extractable phenolics and total extractable tannins in both silages were unaffected by storage temperatures, but condensed tannins in GTS were less when stored at high temperature. The GTS showed no PEG response on in vitro gas production, and revealed only a small increase by PEG on NH3-N concentration. Storage temperature of GTS did not affect the extent of PEG response to both gas production and NH3-N concentration. On the other hand, addition of PEG on BTS markedly increased both the gas production and NH3-N concentration at any ensiled temperature. It can be concluded that tannins in both GTS and BTS suppressed rumen fermentation, and tannins in GTS did more weakly than that in BTS. Ensiling temperature for both tea by-products did not affect the tannin’s activity in the rumen.

  3. THE RUMINANT EFFECT OF VEGETAL LECITHIN AT SHEEP AND GOATS

    Directory of Open Access Journals (Sweden)

    H. SĂRĂNDAN

    2009-05-01

    Full Text Available In the extraction process of the vegetable soy oils and sun-flower oils results in large quantities a waste that contains approximately 45% fat from which 58% is lecithin. This waste called “dreg” creates problems of environment pollution because we didn’t find a use for it. We tested this waste in the food of small ruminants, at sheep and goat, watching the ruminant effect and the apparent digestibility of the nutritive substances in the food. The tested doses of “dregs” were of 100 g and 200 g per day. The food supplementation in sheep and goats with dregs up to 7% fat in the dry substance of the ration has favourable and proportional effects with the dose of fat on the digestibility of the nutritive substances from the food. The growth of ruminant bacteria is favoured at the 100 g dose of dregs but is depressed at the 200 g dose of dregs. On the ruminant protozoa the supplementation with fat from dregs leads to the reducing of the number of protozoa and even at defaunation. It is possible that the fat from the dregs to be a source of YATP and to protect the alimentary proteins of the degrading with proteolytic enzymes and therefore to make the protein ruminant by-pass.

  4. Arginine phosphorylation marks proteins for degradation by a Clp protease.

    Science.gov (United States)

    Trentini, Débora Broch; Suskiewicz, Marcin Józef; Heuck, Alexander; Kurzbauer, Robert; Deszcz, Luiza; Mechtler, Karl; Clausen, Tim

    2016-11-03

    Protein turnover is a tightly controlled process that is crucial for the removal of aberrant polypeptides and for cellular signalling. Whereas ubiquitin marks eukaryotic proteins for proteasomal degradation, a general tagging system for the equivalent bacterial Clp proteases is not known. Here we describe the targeting mechanism of the ClpC-ClpP proteolytic complex from Bacillus subtilis. Quantitative affinity proteomics using a ClpP-trapping mutant show that proteins phosphorylated on arginine residues are selectively targeted to ClpC-ClpP. In vitro reconstitution experiments demonstrate that arginine phosphorylation by the McsB kinase is required and sufficient for the degradation of substrate proteins. The docking site for phosphoarginine is located in the amino-terminal domain of the ClpC ATPase, as resolved at high resolution in a co-crystal structure. Together, our data demonstrate that phosphoarginine functions as a bona fide degradation tag for the ClpC-ClpP protease. This system, which is widely distributed across Gram-positive bacteria, is functionally analogous to the eukaryotic ubiquitin-proteasome system.

  5. Global analysis of phosphorylation and ubiquitylation crosstalk in protein degradation

    Science.gov (United States)

    Swaney, Danielle L.; Beltrao, Pedro; Starita, Lea; Guo, Ailan; Rush, John; Fields, Stanley; Krogan, Nevan J.; Villén, Judit

    2013-01-01

    Crosstalk between different types of post-translational modifications (PTMs) on the same protein molecule adds specificity and combinatorial logic to signal processing, but has not been characterized on a large-scale basis. Here, we developed two methods to identify protein isoforms that are both phosphorylated and ubiquitylated in the yeast Saccharomyces cerevisiae, identifying 466 proteins with 2,100 phosphorylation sites co-occurring with 2,189 ubiquitylation sites. We applied these methods quantitatively to identify phosphorylation sites that regulate protein degradation via the ubiquitin-proteasome system. Our results demonstrate that distinct phosphorylation sites are often used in conjunction with ubiquitylation, and these sites are more highly conserved than the entire set of phosphorylation sites. Finally, we investigated how the phosphorylation machinery can be regulated by ubiquitylation. We found evidence for novel regulatory mechanisms of kinases and 14-3-3 scaffold proteins via proteasome-independent ubiquitylation. PMID:23749301

  6. Ruminal silage degradability and productivity of forage and grain-type sorghum cultivars Degradabilidade ruminal das silagens e produtividade de cultivares de sorgo de tipo forrageiro e granífero

    Directory of Open Access Journals (Sweden)

    José Antônio Resende

    2003-01-01

    Full Text Available Forages for feeding dairy cows should conciliate the potential for high dry matter yield per unit area, maximizing animal stocking rate, with high nutritive value, which enables decreasing the amount of concentrate feedstuffs per milk unit produced. Eighteen sorghum cultivars were cultivated 0.70 m apart and with 12 plants per linear meter. Plants harvested at the milk-to-dough stage were ensiled in the laboratory. Eleven agronomic and chemical traits were evaluated. Silage samples were incubated in situ and the effective ruminal dry matter degradation (DEF was calculated assuming a fractional passage rate of -0.04 h-1. Dry matter yield was 13.0 ± 2.3 (mean ± S.D. t ha-1. The neutral detergent fiber content was 50.3 ± 3.8 and the DEF was 48.8 ± 2.8% (dry matter basis. The average sorghum DEF was 84% of the DEF of a composite sample of 60 corn hybrids simultaneously cultivated, ensiled and ruminally incubated. The 12 forage-type cultivars were taller, more productive, had higher fiber content and lower DEF than the 6 grain and dual purpose-type cultivars. The acid detergent fiber content had the greatest correlation with DEF (r= -0.64. The linear model correlating DEF with productivity was: DEF=54.694 - 0.4449 x t of dry matter ha-1 (r²=0.14. Although there are cultivars that show high productivity and digestibility, it seems to be difficult to conciliate the maximum nutritive value with the maximum productivity of the sorghum crop.Forrageiras para alimentação de vacas leiteiras devem conciliar a alta produção de matéria seca por área, maximizando a taxa de lotação animal, com o alto valor nutritivo, capaz de reduzir a necessidade de alimentos concentrados por litro de leite produzido. Dezoito cultivares de sorgo foram cultivados com espaçamento de 0,70 m e 12 plantas por metro linear. As plantas colhidas ao atingirem o estádio leitoso-pastoso foram ensiladas em laboratório. Onze características agronômicas e químicas foram

  7. FEM1 proteins are ancient regulators of SLBP degradation.

    Science.gov (United States)

    Dankert, John F; Pagan, Julia K; Starostina, Natalia G; Kipreos, Edward T; Pagano, Michele

    2017-03-19

    FEM1A, FEM1B, and FEM1C are evolutionarily-conserved VHL-box proteins, the substrate recognition subunits of CUL2-RING E3 ubiquitin ligase complexes. Here, we report that FEM1 proteins are ancient regulators of Stem-Loop Binding Protein (SLBP), a conserved protein that interacts with the stem loop structure located in the 3' end of canonical histone mRNAs and functions in mRNA cleavage, translation and degradation. SLBP levels are highest during S-phase coinciding with histone synthesis. The ubiquitin ligase complex SCF(cyclin F) targets SLBP for degradation in G2 phase; however, the regulation of SLBP during other stages of the cell cycle is poorly understood. We provide evidence that FEM1A, FEM1B, and FEM1C interact with and mediate the degradation of SLBP. Cyclin F, FEM1A, FEM1B and FEM1C all interact with a region in SLBP's N-terminus using distinct degrons. An SLBP mutant that is unable to interact with all 4 ligases is expressed at higher levels than wild type SLBP and does not oscillate during the cell cycle. We demonstrate that orthologues of SLBP and FEM1 proteins interact in C. elegans and D. melanogaster, suggesting that the pathway is evolutionarily conserved. Furthermore, we show that FEM1 depletion in C. elegans results in the upregulation of SLBP ortholog CDL-1 in oocytes. Notably, cyclin F is absent in flies and worms, suggesting that FEM1 proteins play an important role in SLBP targeting in lower eukaryotes.

  8. Vaccination of Sheep with a Methanogen Protein Provides Insight into Levels of Antibody in Saliva Needed to Target Ruminal Methanogens.

    Science.gov (United States)

    Subharat, Supatsak; Shu, Dairu; Zheng, Tao; Buddle, Bryce M; Kaneko, Kan; Hook, Sarah; Janssen, Peter H; Wedlock, D Neil

    2016-01-01

    Methane is produced in the rumen of ruminant livestock by methanogens and is a major contributor to agricultural greenhouse gases. Vaccination against ruminal methanogens could reduce methane emissions by inducing antibodies in saliva which enter the rumen and impair ability of methanogens to produce methane. Presently, it is not known if vaccination can induce sufficient amounts of antibody in the saliva to target methanogen populations in the rumen and little is known about how long antibody in the rumen remains active. In the current study, sheep were vaccinated twice at a 3-week interval with a model methanogen antigen, recombinant glycosyl transferase protein (rGT2) formulated with one of four adjuvants: saponin, Montanide ISA61, a chitosan thermogel, or a lipid nanoparticle/cationic liposome adjuvant (n = 6/formulation). A control group of sheep (n = 6) was not vaccinated. The highest antigen-specific IgA and IgG responses in both saliva and serum were observed with Montanide ISA61, which promoted levels of salivary antibodies that were five-fold higher than the second most potent adjuvant, saponin. A rGT2-specific IgG standard was used to determine the level of rGT2-specific IgG in serum and saliva. Vaccination with GT2/Montanide ISA61 produced a peak antibody concentration of 7 × 1016 molecules of antigen-specific IgG per litre of saliva, and it was estimated that in the rumen there would be more than 104 molecules of antigen-specific IgG for each methanogen cell. Both IgG and IgA in saliva were shown to be relatively stable in the rumen. Salivary antibody exposed for 1-2 hours to an in vitro simulated rumen environment retained approximately 50% of antigen-binding activity. Collectively, the results from measuring antibody levels and stablility suggest a vaccination-based mitigation strategy for livestock generated methane is in theory feasible.

  9. Probiotic in Ruminant Feed

    Directory of Open Access Journals (Sweden)

    Dicky Pamungkas

    2006-06-01

    Full Text Available The technology development of ruminant feed is related to the effort of fulfilling the nutrient requirement for maintenance and production of rumen microbes and optimizing the protein synthesis of rumen microbes, hence improving the animal production . Probiotic is widely used in feed to avoid the negative effect of antibiotic after therapeutic treatment and to be used as growth promoter . This paper describes the concept of probiotic, selection of microbes for probiotic, the benefit, the effect and the mechanism of probiotic in ruminant . In conclusion, probiotic can improve the animal growth and increase the immunity against diseases .

  10. Design principles of a universal protein degradation machine.

    Science.gov (United States)

    Matyskiela, Mary E; Martin, Andreas

    2013-01-23

    The 26S proteasome is a 2.5-MDa, 32-subunit ATP-dependent protease that is responsible for the degradation of ubiquitinated protein targets in all eukaryotic cells. This proteolytic machine consists of a barrel-shaped peptidase capped by a large regulatory particle, which contains a heterohexameric AAA+ unfoldase as well as several structural modules of previously unknown function. Recent electron microscopy (EM) studies have allowed major breakthroughs in understanding the architecture of the regulatory particle, revealing that the additional modules provide a structural framework to position critical, ubiquitin-interacting subunits and thus allow the 26S proteasome to function as a universal degradation machine for a wide variety of protein substrates. The EM studies have also uncovered surprising asymmetries in the spatial arrangement of proteasome subunits, yet the functional significance of these architectural features remains unclear. This review will summarize the recent findings on 26S proteasome structure and discuss the mechanistic implications for substrate binding, deubiquitination, unfolding, and degradation. Copyright © 2012 Elsevier Ltd. All rights reserved.

  11. 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...

  12. Avaliação da degradabilidade ruminal in situ da palha de arroz fermentada por Trichoderma viride, Rhizopus stolonifer e Pycnoporus sanguineus Evaluation of rice straw fermented by Trichoderma viride, Rhizopus stolonifer and Pycnoporus sanguineus by in situ ruminal degradability

    Directory of Open Access Journals (Sweden)

    Eliane Endres

    1999-08-01

    Full Text Available O melhoramento nutricional da palha de arroz fermentada por Trichoderma viride, Rhizopus stolonifer e Pycnoporus sanguineus foi avaliado por intermédio da degradação ruminal in situ. A fermentação da palha com T. viride e água por duas semanas aumentou a taxa de degradação ruminal em, aproximadamente, 31,5 e 264%, respectivamente, quando comparada com controle e água e com fermentação da palha por uma semana. Houve, também, redução no tempo de degradação efetiva da matéria seca da palha de arroz no trato digestivo do animal. O tratamento com R. stolonifer e água mostrou aumento de 17,95 e 7,90% da fração potencialmente digestível da matéria seca da palha, respectivamente, quando comparada com controle e fermentação por uma semana. O tratamento da palha de arrroz com P. sanguineus e água mostrou redução de 6,66 e 33,64% na degradação ruminal efetiva da matéria seca da palha após fermentação por uma e duas semanas, respectivamente.The nutritional improvement of rice straw fermented by Trichoderma viride, Rhizopus stolonifer and Pycnoporus sanguineus was evaluated using the in situ ruminal degradability technique. The fermentation of the straw with T. viride and water for two weeks increased the degradation rate in 31.5 and 264% when compared with the control and water, and with fermentation of the straw for one week. Also, there was a reduction on the effective degradation time of the straw dry matter in the digestive tract of the animal. The treatment with R. stolonifer and water showed an increase of 17.95 and 7.90% on the potentially digestible fraction of the straw dry matter, respectively, when compared with the control and fermentation by one week. The treatment of straw with P. sanguineus and water showed reduction of 6.66 and 33.64% on the effective ruminal degradability of the straw dry matter after one and two weeks of fermentation, respectively.

  13. Cytoskeletal protein carbonylation and degradation in experimental autoimmune encephalomyelitis

    Science.gov (United States)

    Smerjac, Suzanne M.; Bizzozero, Oscar A.

    2013-01-01

    Protein carbonylation, the non-enzymatic addition of aldehydes or ketones to specific amino acid residues, has been implicated in the pathophysiology of multiple sclerosis (MS). In this study we investigated whether protein carbonyls (PCOs) also accumulate in the spinal cord of Lewis rats with acute experimental autoimmune encephalomyelitis (EAE). Western blots analysis after derivatization with dinitrophenyl hydrazine (oxyblot) showed elevated protein carbonylation at the time of maximal clinical disability. During the same period glutathione levels were substantially reduced, suggesting a causal relationship between these two markers. In contrast, lipid peroxidation products accumulated in EAE spinal cord well before the appearance of neurological symptoms. Carbonyl staining was not restricted to inflammatory lesions but present throughout the spinal cord particularly in neuronal cell bodies and axons. By 2-dimensional-oxyblot we identified several cytoskeletal proteins, including β-actin, GFAP and the neurofilament proteins as the major targets of carbonylation. These findings were confirmed by pull-down experiments, which also showed an increase in the number of carbonylated β-actin molecules and a decrease in that of oxidized neurofilament proteins in EAE. These data suggest the possibility that oxidation targets neurofilament proteins for degradation, which may contribute to axonal pathology observed in MS and EAE. PMID:18088377

  14. In situ identification and quantification of protein-hydrolyzing ruminal bacteria associated with the digestion of barley and corn grain.

    Science.gov (United States)

    Xia, Yun; Kong, Yunhong; Huang, Heping; Yang, Hee Eun; Forster, Robert; McAllister, Tim A

    2016-12-01

    In this study, BODIPY FL DQ™ casein staining combined with fluorescence in situ hybridization (FISH) was used to detect and identify protein-hydrolyzing bacteria within biofilms that produced active cell-surface-associated serine- and metallo-proteases during the ruminal digestion of barley and corn grain in cows fed barley-based diets at 2 different levels. A doublet coccoid bacterial morphotype associated with barley and corn grain particles fluoresced after BODIPY FL DQ™ casein staining. Bacteria with this morphotype accounted for 3%-10% of the total bacteria attached to surface of cereal grain particles, possibly indicative of an important role in the hydrolysis of the protein matrix within the endosperm. However, the identity of these predominant proteolytic bacteria could not be determined using FISH. Quantitative FISH revealed that known proteolytic species, Prevotella ruminicola, Ruminobacter amylophilus, and Butyrivibrio fibrisolvens, were attached to particles of various cultivars of barley grain and corn, confirming their role in the proteolysis of cereal grains. Differences in chemical composition among different barley cultivars did not affect the composition of proteolytic bacterial populations. However, the concentrate level in the basal diet did have an impact on the relative abundance of proteolytic bacteria and thus possibly their overall contribution to the proteolysis of cereal grains.

  15. Accuracy of two optical chlorophyll meters in predicting chemical composition and in vitro ruminal organic matter degradability of Brachiaria hybrid, Megathyrsus maximus, and Paspalum atratum

    Directory of Open Access Journals (Sweden)

    Martin P. Hughes

    2017-03-01

    Full Text Available The objective of this study was to determine the accuracy and reliability of 2 optical chlorophyll meters: FieldScout CM 1,000 NDVI and Yara N-Tester, in predicting neutral detergent fibre (NDF, acid detergent fibre (ADF, acid detergent lignin (ADL, acid detergent insoluble nitrogen (ADIN and in vitro ruminal organic matter degradability (IVOMD of 3 tropical grasses. Optical chlorophyll measurements were taken at 3 stages (4, 8 and 12 weeks of regrowth in Brachiaria hybrid, and Megathyrsus maximus and at 6 and 12 weeks of regrowth in Paspalum atratum (cv. Ubon. Optical chlorophyll measurements showed the highest correlation (r = 0.57 to 0.85 with NDF concentration. The FieldScout CM 1,000 NDVI was better than the Yara N-Tester in predicting NDF (R2 = 0.70 and ADF (R2 = 0.79 concentrations in Brachiaria hybrid and NDF (R2 = 0.79 in M. maximus. Similarly, FieldScout CM 1,000 NDVI produced better estimates of 24 h IVOMD (IVOMD24h in Brachiaria hybrid (R2 = 0.81 and IVOMD48h in Brachiaria hybrid (R2 = 0.65 and M. maximus (R2 = 0.75. However, these prediction models had relatively low concordance correlation coefficients, i.e., CCC >0.90, but random errors were the main source of bias. It was, therefore, concluded that both optical chlorophyll meters were poor and unreliable predictors of ADIN and ADL concentrations. Overall, the FieldScout CM 1,000 NDVI shows potential to produce useful estimates of IVOMD24h and ADF in Brachiaria hybrid and IVOMD48h and NDF concentrations in M. maximus.

  16. Effects of extruding wheat dried distillers grains with solubles with peas or canola meal on ruminal fermentation, microbial protein synthesis, nutrient digestion, and milk production in dairy cows.

    Science.gov (United States)

    Claassen, R M; Christensen, D A; Mutsvangwa, T

    2016-09-01

    Our objective was to examine the effects of feeding coextruded and nonextruded supplements consisting of wheat dried distillers grains with solubles with peas (WDDGS-peas) or canola meal (WDDGS-CM) on ruminal fermentation, omasal flow, and production performance in Holstein cows. Eight cows (4 ruminally cannulated) were used in a replicated 4×4 Latin square with 28-d periods and a 2×2 factorial arrangement of dietary treatments. Dietary treatments were coextruded or nonextruded mixtures of WDDGS-peas and WDDGS-CM that were included in total mixed rations at 15.1% [dry matter (DM) basis]. Diet had no effect on DM intake. Milk yield was greater in cows fed coextruded diets compared with those fed nonextruded diets. Milk fat content was greater in cows fed nonextruded diets compared with those fed coextruded diets, but milk fat yield was greater in cows fed coextruded diets compared with those fed nonextruded diets. Milk yield tended to be greater and milk protein yield was greater in cows fed WDDGS-peas compared with those fed WDDGS-CM. Cows fed nonextruded diets had a greater milk urea-N concentration compared with those fed coextruded diets. Cows fed coextruded diets had greater ruminal digestion of DM and tended to have greater ruminal digestion of organic matter compared with those fed nonextruded diets. Total-tract digestibilities of organic matter, crude protein, ether extract, and starch were greater, whereas that of acid detergent fiber and neutral detergent fiber tended to be greater in cows fed coextruded compared with those fed nonextruded diets. Total-tract digestibility of ether extract was lower whereas that of starch was greater and that of crude protein tended to be greater in cows fed WDDGS-peas compared with those fed WDDGS-CM. Total N excretion and milk N efficiency were unaffected by diet. Ruminal NH3-N concentration tended to be greater in cows fed WDDGS-CM compared with those fed WDDGS-peas. Ruminal propionate concentration was greater whereas

  17. CHIP Regulates Osteoclast Formation through Promoting TRAF6 Protein Degradation

    Science.gov (United States)

    Li, Shan; Shu, Bing; Zhang, Yanquan; Li, Jia; Guo, Junwei; Wang, Yinyin; Ren, Fangli; Xiao, Guozhi; Chang, Zhijie; Chen, Di

    2014-01-01

    Objective Carboxyl terminus of Hsp70-interacting protein (CHIP or STUB1) is an E3 ligase and regulates the stability of several proteins which are involved in tumor growth and metastasis. However, the role of CHIP in bone growth and bone remodeling in vivo has not been reported. The objective of this study is to investigate the role and mechanism of CHIP in regulation of bone mass and bone remodeling. Methods The bone phenotype of Chip−/− mice was examined by histology, histomorphometry and micro-CT analyses. The regulatory mechanism of CHIP on the degradation of TRAF6 and the inhibition of NF-κB signaling was examined by immunoprecipitation (IP), western blotting and luciferase reporter assays. Results In this study, we found that deletion of the Chip gene leads to osteopenic phenotype and increased osteoclast formation. We further found that TRAF6, as a novel substrate of CHIP, is up-regulated in Chip−/− osteoclasts. TRAF6 is critical for RANKL-induced osteoclastogenesis. TRAF6 is an adaptor protein which functions as an E3 ligase to regulate the activation of TAK1 and the I-κB kinase (IKK) and is a key regulator of NF-κB signaling. CHIP interacts with TRAF6 to promote TRAF6 ubiquitination and proteasome degradation. CHIP inhibits p65 nuclear translocation, leading to the repression of the TRAF6-mediated NF-κB transcription. Conclusion CHIP inhibits NF-κB signaling via promoting TRAF6 degradation and plays an important role in osteoclastogenesis and bone remodeling, suggesting that it may be a novel therapeutic target for the treatment of bone loss associated diseases. PMID:24578159

  18. Role of phosphorolytic cleavage in cellobiose and cellodextrin metabolism by the ruminal bacterium Prevotella ruminicola.

    OpenAIRE

    Lou, J.; Dawson, K A; Strobel, H J

    1996-01-01

    In bacteria, cellobiose and cellodextrins are usually degraded by either hydrolytic or phosphorolytic cleavage. Prevotella ruminicola B(1)4 is a noncellulolytic ruminal bacterium which has the ability to utilize the products of cellulose degradation. In this organism, cellobiose hydrolytic cleavage activity was threefold greater than phosphorolytic cleavage activity (113 versus 34 nmol/min/mg of protein), as measured by an enzymatic assay. Cellobiose phosphorylase activity (measured as the re...

  19. Effects of dietary nitrogen concentration on messenger RNA expression and protein abundance of urea transporter-B and aquaporins in ruminal papillae from lactating Holstein cows

    DEFF Research Database (Denmark)

    Røjen, Betina Amdisen; Poulsen, Søren Brandt; Theil, Peter Kappel

    2011-01-01

    lactating dairy cows. Ruminal papillae were harvested from cows fed low N (12.9% crude protein) and high N (17.1% crude protein) diets in a crossover design with 21-d periods. The mRNA expression was determined by real-time reverse transcription-PCR and protein abundance by immunoblotting. The m......RNA expression of UT-B was not affected by dietary treatment, whereas mRNA expression of AQP3, 7, and 10 were greater in the high N compared with the low N fed cows. Using peptide-derived rabbit antibodies to cow AQP3, 7, and 8, immunoblotting revealed bands of approximately 27, 27, and 24 kDa in ruminal...... papillae, respectively. A peptide-derived chicken antibody to cow UT-B detected a band of approximately 30 to 32 kDa in ruminal papillae. The abundance of UT-B and AQP3 and 7 were not affected by dietary treatment. In contrast, the abundance of AQP8 was greater in high N compared with low N diets...

  20. How is the balance between protein synthesis and degradation achieved?

    Directory of Open Access Journals (Sweden)

    Rothman Stephen

    2010-06-01

    Full Text Available Abstract Unlike most substances that cells manufacture, proteins are not produced and broken down by a common series of chemical reactions, but by completely different (independent and disconnected mechanisms that possess no intrinsic means of making the rates of the two processes equal and attaining steady state concentrations. Balance between them is achieved extrinsically and is often imagined today to be the result of the actions of chemical feedback agents. But however instantiated, chemical feedback or any similar mechanism can only rectify induced imbalances in a system previously balanced by other means. Those "other means" necessarily involve reversible mass action or equilibrium-based interactions between native and altered forms of protein molecules somewhere in time and space between their synthesis and degradation.

  1. Vaccination of Sheep with a Methanogen Protein Provides Insight into Levels of Antibody in Saliva Needed to Target Ruminal Methanogens.

    Directory of Open Access Journals (Sweden)

    Supatsak Subharat

    Full Text Available Methane is produced in the rumen of ruminant livestock by methanogens and is a major contributor to agricultural greenhouse gases. Vaccination against ruminal methanogens could reduce methane emissions by inducing antibodies in saliva which enter the rumen and impair ability of methanogens to produce methane. Presently, it is not known if vaccination can induce sufficient amounts of antibody in the saliva to target methanogen populations in the rumen and little is known about how long antibody in the rumen remains active. In the current study, sheep were vaccinated twice at a 3-week interval with a model methanogen antigen, recombinant glycosyl transferase protein (rGT2 formulated with one of four adjuvants: saponin, Montanide ISA61, a chitosan thermogel, or a lipid nanoparticle/cationic liposome adjuvant (n = 6/formulation. A control group of sheep (n = 6 was not vaccinated. The highest antigen-specific IgA and IgG responses in both saliva and serum were observed with Montanide ISA61, which promoted levels of salivary antibodies that were five-fold higher than the second most potent adjuvant, saponin. A rGT2-specific IgG standard was used to determine the level of rGT2-specific IgG in serum and saliva. Vaccination with GT2/Montanide ISA61 produced a peak antibody concentration of 7 × 1016 molecules of antigen-specific IgG per litre of saliva, and it was estimated that in the rumen there would be more than 104 molecules of antigen-specific IgG for each methanogen cell. Both IgG and IgA in saliva were shown to be relatively stable in the rumen. Salivary antibody exposed for 1-2 hours to an in vitro simulated rumen environment retained approximately 50% of antigen-binding activity. Collectively, the results from measuring antibody levels and stablility suggest a vaccination-based mitigation strategy for livestock generated methane is in theory feasible.

  2. RUMINAL DEGRADATION KINETIC PARAMETERS OF COFFEE HULLS (Coffea arabica, L. TREATED WITH SODIUM HYDROXIDE (NAOH PARÂMETROS CINÉTICOS DA DEGRADAÇÃO RUMINAL DA CASCA DE CAFÉ (Coffea arabica, L. TRATADA COM HIDRÓXIDO DE SÓDIO (NAOH

    Directory of Open Access Journals (Sweden)

    Luciano Fernandes de Sousa

    2008-04-01

    Full Text Available The objective of this work was to evaluate dry matter ruminal degradation kinetics of coffee hulls treated with increasing sodium hydroxide quantities. Two rumen fistulated cows were used to incubate samples in nylon bags for 12. 24, 36. 48 and 72 hours. Four ruminal incubation periods were used, in a complete randomized block design. Coffee hulls were treated with 0%, 3%, 6% and 9% of sodium hydroxide (dry matter basis, corresponding to treatments T1 to t4 respectively. Experimental results were compared using Tukey test, at 5% probability level, as follows for treatments 1 to 4 respectively: soluble fraction (9.35d; 17.65c; 31.93b; 32.28a, de (34.40d; 40.50c; 43.28b; 50.35a, potential degradability (44.33d; 50.33c; 52.35b; 57.70a and lag time in hours (4.03a; 3.93a; 4.33a; 2.55a. The results indicate that increasing the levels of NaOH in the coffee hulls treatments increased significantly their ruminal solubility, as well as their effective and potential degradabilities. However that increase had no effect upon coffee hulls lag time in the rumen.KEY WORDS: by-product feedstuffs, rumen degradability, ruminant.

    O objetivo deste trabalho foi o de determinar a cinética de degradação ruminal da matéria seca da casca de café, tratada com diferentes quantidades de hidróxido de sódio. Utilizaram-se duas vacas fistuladas no rúmen, incubando-se as amostras em sacolas de náilon por 12, 24, 36, 48 e 72 horas, por quatro rodadas seqüenciais, sendo que cada uma destas representou um bloco, dentro de um delineamento de blocos inteiramente casualizados. Tratou-se a casca de café com 0%, 3%, 6% e 9 % de hidróxido de sódio (base seca constituindo assim os tratamentos t1

  3. A comparison of different legume seeds as protein supplement to optimise the use of low quality forages by ruminants.

    Science.gov (United States)

    Yáñez-Ruiz, David R; Martín-García, Antonio I; Weisbjerg, Martin R; Hvelplund, Torben; Molina-Alcaide, Eduarda

    2009-02-01

    The potential of different legume seeds species, including recently new developed varieties (Vicia faba: a commercial variety and varieties Alameda, Palacio and Baraka; Lupinus angustifolius; Pisum sativum and Cicer arietinum: varieties Fardon and Zegri) as protein supplements to low quality forages was evaluated. First, the chemical composition, in vitro digestibility, in situ degradability and in vitro/in situ intestinal digestibility of legume seeds were determined. The chemical composition was rather similar within genus. Vicia faba beans contained more condensed tannins (35.8-56.4 g/kg DM) and less ether extract (12.8-9.5 g/kg DM) than the other legumes. The rumen degradability and intestinal digestibility of the undegraded protein in the rumen was very similar among the seeds, with exception of lupins, having a much higher degradation rate than the rest. Second, the fermentation characteristics of diets, which were based on two low quality forages (olive leaves and barley straw) and feed blocks without or with supplementation of legume seeds or soybean meal, were investigated by using single-flow continuous culture fermenters. In this trial, the fermentation parameters (ammonia, pH and volatile fatty acids), the microbial protein synthesis and the degradation of olive leaves and barley straw promoted by the different diets were studied. Compared to soybean meal, beans and peas showed similar suitability as protein supplements for sustaining in vitro fermentation of low-quality forages. However, our results suggest a significant interaction between the type of legume used as supplement and the type of forage used, which need to be further studied in vivo.

  4. Parâmetros ruminais e síntese de proteína metabolizável em bovinos de corte sob suplementação com proteinados contendo diversos níveis de proteína bruta Ruminal fermentation characteristics and protein fraction effects on metabolizable protein synthesis of beef cattle fed different levels of crude protein

    Directory of Open Access Journals (Sweden)

    Luiz Orcirio Fialho de Oliveira

    2009-12-01

    , ruminal undegradable protein (RUP and endogenous crude protein (ECP and their contributions to the pool of metabolizable protein (MP. Four Nelore steer, 395 ± 9 kg, fitted with ruminal cannulas, were used to evaluate ruminal parameters, degradability, ruminal kinetics and microbial synthesis in a 4 ×4 Latin square design. The animals were supplied with 400g/head/day of supplements containing 30, 40, and 50% crude protein (CP for comparison with a control group without protein supplementation (C. The animals grazed Brachiaria brizantha cv Marandu, distributed in four one-hectare paddocks where the supplements were offered and the orts removed daily. The N-NH3 levels in the animals that received 50% CP were higher than those observed in animals receiving 40% CP and C, but were similar to the levels observed in the animals supplemented with 30% CP. The VFA concentrations in the group supplemented with 30% PB were higher than the control treatment (C and similar to those obtained with 40 and 50% CP supplementation. The pH did not differ among the groups. The microbial synthesis and RUP were greater for the animals that received protein supplementation compared to the control treatment.

  5. Ruminal degradability of hard or soft texture corn grain at three maturity stages Degradabilidade ruminal de grãos de milho de textura dura ou macia em três estádios de maturação

    Directory of Open Access Journals (Sweden)

    Marcos Neves Pereira

    2004-01-01

    Full Text Available The predominance of vitreous endosperm in hard texture flint corn (Zea mays L. can decrease ruminal starch digestion comparatively to the farinaceous endosperm of dent corn, reducing energy content of the grain. The objective of this experiment was to evaluate the effects of texture and maturity stage on ruminal degradability of corn grain. Two dent and two flint hybrids were harvested at the early dent, half milk line, and black layer stages. The proportion of vitreous endosperm (vitreousness in dent hybrids was 44.3%, while in flint it was 67.0%. There was a linear increase in vitreousness with advancing maturity. Flint hybrids at the early dent stage were more vitreous than dent at the black layer stage. The increase in vitreousness per maturation day was greater for flint hybrids. Grains were incubated in situ in the rumen of 6 cows. The 24-hour ruminal dry matter degradation was 63.3% for dent corn and 52.4% for flint corn. The 72-hour incubation residues of dent and flint hybrids were 7.6% and 15.6%, respectively. Ruminal degradability was similar between hybrids at the early dent and half milk line stages. There was a marked texture effect on ruminal degradability at the black layer stage (quadratic effect of maturity stage and interaction between texture and maturity stage. Use of dent hybrids, compared to flint hybrids, may result in smaller relative reduction in ruminal starch digestion in situations of late grain harvesting.A predominância de endosperma vítreo em milho (Zea mays L. flint de textura dura pode deprimir a digestão ruminal do amido comparativamente ao endosperma farináceo de milho dentado, reduzindo o conteúdo energético do grão. O objetivo deste experimento foi avaliar os efeitos da textura e do estádio de maturidade sobre a degradabilidade ruminal de grãos de milho. Dois híbridos dentados e dois duros foram colhidos nos estádios dentado inicial, metade da linha do leite e linha preta. A proporção de

  6. Níveis de substituição da silagem de milho pela silagem de resíduos industriais de abacaxi sobre a degradabilidade ruminal em bovinos de corte Substitution levels of corn silage by pineapple by-products on ruminal degradability in beef cattle

    Directory of Open Access Journals (Sweden)

    Fabiana Helena Lallo

    2003-06-01

    Full Text Available Objetivou-se avaliar a cinética de degradação ruminal e a degradabilidade efetiva (DE da matéria seca (MS e proteína bruta (PB de quatro rações experimentais, com diferentes níveis de substituição (0, 20, 40 e 60% da silagem de milho pela silagem de resíduos industriais de abacaxi e da silagem de resíduos industriais de abacaxi individualmente. Foram utilizados três bovinos inteiros (½ Nelore x ½ Angus, portadores de cânula ruminal, com peso vivo médio de 350 kg. As amostras das rações e da silagem de resíduos industriais de abacaxi individualmente foram homogeneizadas e moídas em moinho com peneira de 5 mm de crivo. Os alimentos foram incubados no rúmen em sacos de náilon nos tempos 4, 8, 12, 24, 48, 72 e 96 horas. A silagem de resíduos industriais de abacaxi incubada individualmente apresentou valores de 28,8%; 49,4% e 4,2%/h para os parâmetros a, b e c da MS e 55,9%; 34,6% e 5,5%/h para PB. Ainda, apresentou valores de 51,4 e 74,1%, respectivamente para DE da MS e PB para a taxa de passagem estimada em 5%/h. Para a MS, a substituição da silagem de milho pela silagem de resíduos industriais de abacaxi, mostrou comportamento quadrático negativo e para a fração a e quadrático positivo para a fração b. O parâmetro c apresentou comportamento quadrático, com valor máximo para o T40. Para a PB, o parâmetro a apresentou comportamento linear positivo e o parâmetro c linear negativo. Ainda, o parâmetro b da PB apresentou comportamento quadrático positivo. Para a taxa de passagem de 5%/h, a DE da MS foi de 53,0; 51,9; 55,2 e 50,9% e da PB foi de 70,7; 70,5; 66,8 e 72,7%, para os níveis de substituição de 0, 20, 40 e 60% da silagem de milho pela silagem de resíduos industriais de abacaxi.The objective of this experiment was evaluate the ruminal degradation kinects and effective degradability (ED of dry matter (DM and crude protein (CP of four experimental diets, with different substitution levels (0, 20, 40

  7. Growth-based determination and biochemical confirmation of genetic requirements for protein degradation in Saccharomyces cerevisiae.

    Science.gov (United States)

    Watts, Sheldon G; Crowder, Justin J; Coffey, Samuel Z; Rubenstein, Eric M

    2015-02-16

    Regulated protein degradation is crucial for virtually every cellular function. Much of what is known about the molecular mechanisms and genetic requirements for eukaryotic protein degradation was initially established in Saccharomyces cerevisiae. Classical analyses of protein degradation have relied on biochemical pulse-chase and cycloheximide-chase methodologies. While these techniques provide sensitive means for observing protein degradation, they are laborious, time-consuming, and low-throughput. These approaches are not amenable to rapid or large-scale screening for mutations that prevent protein degradation. Here, a yeast growth-based assay for the facile identification of genetic requirements for protein degradation is described. In this assay, a reporter enzyme required for growth under specific selective conditions is fused to an unstable protein. Cells lacking the endogenous reporter enzyme but expressing the fusion protein can grow under selective conditions only when the fusion protein is stabilized (i.e. when protein degradation is compromised). In the growth assay described here, serial dilutions of wild-type and mutant yeast cells harboring a plasmid encoding a fusion protein are spotted onto selective and non-selective medium. Growth under selective conditions is consistent with degradation impairment by a given mutation. Increased protein abundance should be biochemically confirmed. A method for the rapid extraction of yeast proteins in a form suitable for electrophoresis and western blotting is also demonstrated. A growth-based readout for protein stability, combined with a simple protocol for protein extraction for biochemical analysis, facilitates rapid identification of genetic requirements for protein degradation. These techniques can be adapted to monitor degradation of a variety of short-lived proteins. In the example presented, the His3 enzyme, which is required for histidine biosynthesis, was fused to Deg1-Sec62. Deg1-Sec62 is targeted for

  8. Cytokinin inhibits the proteasome-mediated degradation of carbonylated proteins in Arabidopsis leaves

    Science.gov (United States)

    Under normal conditions, plants contain numerous carbonylated proteins, which are thought to be indicative of oxidative stress damage. Conditions that promote formation of reactive oxygen species (ROS) enhance protein carbonylation, and protein degradation is required to reverse the damage. However,...

  9. Effects of feeding corn silage inoculated with microbial additives on the ruminal fermentation, microbial protein yield, and growth performance of lambs.

    Science.gov (United States)

    Basso, F C; Adesogan, A T; Lara, E C; Rabelo, C H S; Berchielli, T T; Teixeira, I A M A; Siqueira, G R; Reis, R A

    2014-12-01

    This study aimed to examine the effects of feeding corn silage inoculated without or with either Lactobacillus buchneri (LB) alone or a combination of LB and Lactobacillus plantarum (LBLP) on the apparent digestibility, ruminal fermentation, microbial protein synthesis, and growth performance of lambs. Thirty Santa Inês×Dorper crossbred intact males lambs weighing 20.4±3.8 kg were blocked by weight into 10 groups. Lambs in each group were randomly assigned to 1 of the following 3 dietary treatments: untreated (Control), LB, and LBLP silage. Lambs were fed experimental diets for 61 d. The apparent digestibility was indirectly estimated from indigestible NDF measured on d 57 to 59. Spot urine samples were collected from all animals on d 59 to estimate microbial protein synthesis. Lambs were slaughtered for carcass evaluation on d 61 when they weighed 32.4±5.2 kg. Six additional ruminally cannulated Santa Inês×Dorper crossbred wethers weighing 40.5±1.8 kg were used to examine dietary effects on ruminal fermentation. Average daily gain was increased when lambs were fed LBLP silage (Psilage. The LBLP silage had the highest (Psilages had greater acetic acid concentrations than the Control silage (Psilage increased intakes of DM, OM, CP, NDF, total carbohydrate (CHO), and GE by the lambs but decreased digestibility of DM, OM, CP, total and nonstructural carbohydrates, and concentration of GE and ME. (Psilages had greater microbial N supply than those on the Control treatment (Psilage fermentation. The intakes of DM, OM, CP, NDF, and GE were improved in the lambs fed corn silage inoculated with L. buchneri alone or combined with L. plantarum. The microbial N supply was enhanced in the lambs fed corn silage inoculated with L. buchneri. The inoculation of L. buchneri combined with L. plantarum reduced the acetate to propionate ratio in ruminal fluid and improved the ADG of lambs.

  10. Post-ruminal digestibility of crude protein from grass and grass silages in cows

    NARCIS (Netherlands)

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

    2006-01-01

    Grass samples were grown on a clay or sandy soil, fertilised with 150 or 300 kg N/ha per year, and harvested on different days during two consecutive growing seasons. The grass samples were stored frozen or ensiled after wilting to approximately 250 or 450 g DM/kg. The recoveries of crude protein

  11. Daily and alternate-day supplementation of urea or biuret to ruminants consuming low-quality forage: III. Effects on ruminal fermentation characteristics in steers.

    Science.gov (United States)

    Currier, T A; Bohnert, D W; Falck, S J; Schauer, C S; Bartle, S J

    2004-05-01

    Five ruminally and duodenally cannulated steers (491 +/- 21 kg BW) were used in an incomplete 5 x 4 Latin square with four 24-d periods to determine the influence of supplemental nonprotein N (NPN) source and supplementation frequency (SF) on the dynamics of ruminal fermentation in steers consuming low-quality grass straw (4% CP). Treatments (TRT) included an unsupplemented control (CON) and a urea or biuret supplement that were placed directly into the rumen at 0700 daily (D) or every other day (2D). The NPN treatments were formulated to provide 90% of the estimated degradable intake protein requirement; therefore, the urea and biuret treatments received the same amount of supplemental N over a 2-d period. Daily TRT were supplemented with CP at 0.04% of BW/d, whereas the 2D TRT were supplemented at 0.08% of BW every other day. Forage was provided at 120% of the previous 5-d average intake in two equal portions at 0715 and 1900. Ruminal fluid was collected 0, 3, 6, 9, 12, and 24 h after supplementation on a day of and a day before supplementation for all TRT. Ruminal NH3-N increased (P biuret. Ruminal NH3-N on the day only daily supplements were provided was greater (P = 0.02) for D compared with 2D. On the day all supplements were provided, D increased (P = 0.05) ruminal indigestible acid detergent fiber passage rate and ruminal fluid volume compared with 2D. These results suggest that urea or biuret can be used effectively as a supplemental N source by steers consuming low-quality forage without adversely affecting ruminal fermentation, even when provided every other day.

  12. Rumen epithelial adaptation to ruminal acidosis in lactating cattle involves the coordinated expression of insulin-like growth factor-binding proteins and a cholesterolgenic enzyme.

    Science.gov (United States)

    Steele, M A; Dionissopoulos, L; AlZahal, O; Doelman, J; McBride, B W

    2012-01-01

    The objective of this study was to characterize the mRNA expression of metabolic and proliferative genes in the rumen epithelium during ruminal acidosis. To meet our objectives, 16 rumen-fistulated, lactating Holstein dairy cattle (618±35 kg of body weight, 221±32 d in milk) were used in a randomized complete block design. All cattle were fed a high-forage diet (HF; 88.9% of dry matter) for 5 wk before the experiment. After the baseline week (wk 0), half of the cattle were randomly assigned and transitioned to a high-concentrate diet (HC; 62.2% of dry matter) which was fed for 3 wk (wk 1, 2, and 3). For the last 48 h of each week, continuous ruminal pH, short-chain fatty acids, and plasma β-hydroxybutyrate were assessed, followed by a rumen papillae biopsy. Milk production was higher in HC cattle compared with HF during wk 1, 2, and 3 (17.4±0.5 vs. 23.4±0.9 kg/d, respectively); however, the mean ruminal pH was decreased (5.75±0.03 vs. 6.30±0.02). The HC cattle spent more time below pH 5.6 (594±54 vs. 3±3 min/d) and displayed greater concentrations of ruminal butyrate (15.8±0.9 vs. 10.2±0.4 mmol) and plasma β-hydroxybutyrate (1,036±63 vs. 778±20 μM) compared with the HF cattle. The mRNA expression of genes involved in ketogenesis (HMGCS2 and PPARA) and short-chain fatty acid transport (MCT1) was unchanged by treatment. However, a downregulation in HMGCS1 (0.72±0.09), one of the cholesterol biosynthesis genes, was observed in HC cattle during wk 1 of the grain challenge. In addition, the relative mRNA expression value of insulin-like growth factor-binding protein 3 was lower (0.78±0.06), whereas insulin-like growth factor-binding protein 5 was higher (1.79±0.15) in HC compared with HF cattle. These results suggest that grain-induced ruminal acidosis alters the mRNA expression of IGF-binding proteins and a cholesterolgenic enzyme in the rumen epithelium of lactating dairy cattle. Copyright © 2012 American Dairy Science Association. Published by

  13. Incomplete proteasomal degradation of green fluorescent proteins in the context of tandem fluorescent protein timers.

    Science.gov (United States)

    Khmelinskii, Anton; Meurer, Matthias; Ho, Chi-Ting; Besenbeck, Birgit; Füller, Julia; Lemberg, Marius K; Bukau, Bernd; Mogk, Axel; Knop, Michael

    2016-01-15

    Tandem fluorescent protein timers (tFTs) report on protein age through time-dependent change in color, which can be exploited to study protein turnover and trafficking. Each tFT, composed of two fluorescent proteins (FPs) that differ in maturation kinetics, is suited to follow protein dynamics within a specific time range determined by the maturation rates of both FPs. So far, tFTs have been constructed by combining slower-maturing red fluorescent proteins (redFPs) with the faster-maturing superfolder green fluorescent protein (sfGFP). Toward a comprehensive characterization of tFTs, we compare here tFTs composed of different faster-maturing green fluorescent proteins (greenFPs) while keeping the slower-maturing redFP constant (mCherry). Our results indicate that the greenFP maturation kinetics influences the time range of a tFT. Moreover, we observe that commonly used greenFPs can partially withstand proteasomal degradation due to the stability of the FP fold, which results in accumulation of tFT fragments in the cell. Depending on the order of FPs in the timer, incomplete proteasomal degradation either shifts the time range of the tFT toward slower time scales or precludes its use for measurements of protein turnover. We identify greenFPs that are efficiently degraded by the proteasome and provide simple guidelines for the design of new tFTs. © 2016 Khmelinskii 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).

  14. The Advantages of Targeted Protein Degradation Over Inhibition: An RTK Case Study.

    Science.gov (United States)

    Burslem, George M; Smith, Blake E; Lai, Ashton C; Jaime-Figueroa, Saul; McQuaid, Daniel C; Bondeson, Daniel P; Toure, Momar; Dong, Hanqing; Qian, Yimin; Wang, Jing; Crew, Andrew P; Hines, John; Crews, Craig M

    2017-10-25

    Proteolysis targeting chimera (PROTAC) technology has emerged over the last two decades as a powerful tool for targeted degradation of endogenous proteins. Herein we describe the development of PROTACs for receptor tyrosine kinases, a protein family yet to be targeted for induced protein degradation. The use of VHL-recruiting PROTACs against this protein family reveals several advantages of degradation over inhibition alone: direct comparisons of fully functional, target-degrading PROTACs with target-inhibiting variants that contain an inactivated E3 ligase-recruiting ligand show that degradation leads to more potent inhibition of cell proliferation and a more durable and sustained downstream signaling response, and thus addresses the kinome rewiring challenge seen with many receptor tyrosine kinase inhibitors. Combined, these findings demonstrate the ability to target receptor tyrosine kinases for degradation using the PROTAC technology and outline the advantages of this degradation-based approach. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. 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

    The Dutch feed evaluation system for ruminants uses assumptions and regression equations to estimate the intestinal digestibility of crude protein (CP), neutral detergent fibre (NDF) and starch. These assumptions and equations are based on many different studies, obtained over a very long period....... and Protein System; CP, crude protein; DM, dry matter; DRUP, intestinal digestion of rumen undegraded feed protein; Fim, Feed into Milk system; NDF, neutral detergent fibre; RUP, rumen undegraded protein; SD, standard deviation; TMR, total mixed ration...

  16. 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 ...

  17. Effect of Rapeseed Oil on Effective Protein Degradability and Intestinal Protein Digestibility of Oat, Rapeseed Meal and Dried Sugarbeet Pulp

    National Research Council Canada - National Science Library

    Potkanski, A; Nowak, W

    2000-01-01

    Potkanski, A. and Nowak, W. 2000. Effect of rapeseed oil on effective protein degradability and intestinal protein digestibility of oat, rapeseed meal and dried sugarbeet pulp. J. Appl. Anim. Res., 18: 81-89...

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

    Directory of Open Access Journals (Sweden)

    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.

  19. Determination of Cell Wall Protein from Selected Feedstuffs and its Relationship with Ruminal Protein Digestibility in Vitro

    Directory of Open Access Journals (Sweden)

    Anuraga Jayanegara

    2016-08-01

    Full Text Available This study was aimed to analyze neutral detergent insoluble crude protein (NDICP and acid detergent insoluble crude protein (ADICP contents of various commonly used forage and concentrate feedstuffs in Indonesia. A number of forages and concentrates, i.e. gliricidia, trichantera, indigofera, calliandra, papaya leaves, cassava leaves, leucaena, rapeseed meal, corn gluten feed, soybean meal, copra meal, palm kernel meal, fish and bone meal and wheat bran were subjected to proximate analysis, cell wall nitrogen determination and in vitro rumen fermentation evaluation. Chemical composition analysis was done in duplicate. The in vitro incubation was conducted in 14 treatments and 3 replicates by following a randomized complete block design. Variables measured after the incubation were total volatile fatty acid (VFA, ammonia, dry matter digestibility (DMD, organic matter digestibility (OMD and crude protein digestibility (CPD. Results revealed that among the forages, cassava leaf had the highest proportion of NDICP while corn gluten feed was the highest among the concentrates. Different from that of NDICP, the highest proportion of ADICP was obtained in papaya leaf and copra meal for the forages and concentrates, respectively. Higher proportion of NDICP tended to decrease CPD (P<0.1. Although higher ADICP apparently tended to decrease CPD as well, the relationship was insignificant due to the higher variation of the dependent variable. It was concluded that cell wall protein proportion in feed may be used as an indicator to determine the quality of protein and its utilization in the rumen.

  20. 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 results showed that increased degradability of the protein in the rumen decreased the intestinal digestibility, which shows that feedstuffs contain a protein fraction which is both undegradable in the rumen and indigestible in the intestine. The results thus indicate that it is possible to calculate...... the intestinal digestibility from information on the intestinal digestibility of the protein in the intact feed at any degradability estimated. The results clearly show that intestinal digestibility of undegraded dietary protein cannot be considered as a constant value as used in most protein evaluation systems...

  1. 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.

  2. Degradation of two soluble proteins--casein and egg protein by a macro in vitro method.

    Science.gov (United States)

    Udén, P

    2013-08-01

    Degradation of casein and egg protein was studied with whole rumen contents (RC) in a macro in vitro system to elucidate previous findings of initial rapid disappearance of soluble proteins in vitro. Five to 7.5 kg of RC from a dry and/or a lactating cow were incubated with buffer and casein or egg protein for 180 min with frequent sampling. Degradation was measured as loss of trichloroacetic acid precipitable N (TCA-N) from the inocula. Normal (39°C) and low (2°C) temperature incubations were examined in Exp. 1, using 1 g of TCA-N from casein. Four levels of casein (0-12 g TCA-N) in Exp. 2 and four levels of egg albumin (0-24 g TCA-N) in Exp. 3 were fermented at 39°C. Initial recovery of casein TCA-N was 106% at 2°C and 56% at 39°C (Exp. 1). Casein (TCA-N) recovered initially increased in Exp. 2 from 21% at 3 g to 86% at 12 g TCA-N, while absolute loss remained relatively constant at 358 mg TCA-N/kg RC (SD=47). Fractional degradation rate was highest (0.03/min) at the intermediate dosage level. In the absence of rumen fluid (Exp. 4), no casein was lost. Initial egg protein recovery was on average 103% (Exp. 3). Recovery seemed unaffected by dosage level, and absolute degradation rate was relatively constant over time and increased with dosage level (p<0.001) from 1.48 to 2.95 mg TCA-N/(kg RC × min). Maximum degradation rate [mg TCA-N/(kg RC × min)] and affinity constant (mg TCA-N/kg RC) were estimated at 261 and 1650, respectively. It is concluded that a surprisingly constant amount of casein disappears immediately from warm rumen fluid and that this does not occur either with chilled RC, in the absence of rumen fluid, or when replaced with egg protein. The mechanisms for this disappearance are yet to be discovered. © 2012 Blackwell Verlag GmbH.

  3. 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. Rec...

  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...

  5. Degradabilidade e produção de gases in vitro de fontes energéticas alternativas na alimentação de ruminantes = Degradability and in vitro gas production of alternative energy sources in ruminant feed

    Directory of Open Access Journals (Sweden)

    Roberto de Camargo Wascheck

    2010-10-01

    Full Text Available O padrão fermentativo in vitro de dietas compostas por fubá de milho, farelo de arroz integral, farelo de arroz parboilizado e suas associações em diferentes proporções foi estudado, na porção concentrada da dieta de ruminantes. Até 12h de incubação, a produçãode gases foi superior (p The in vitro fermentation pattern of diets composed of corn meal, rice bran, parboiled rice bran and their associations was studied in different proportions in the concentrate portion in ruminants diets. Up to 12 hours of incubation, gas production was higher (p < 0.05 for the treatments with rice bran (FAI and parboiled rice bran (FAP; however, after 96 hours, the treatment with 100% corn meal (FMproduced more (p < 0.05 gases (379.41 mL g DM-1. Total degradability of dry matter (DM and organic matter (OM were higher (p < 0.05 for treatments FAI and FAP up to 12 hours; however, after 96 hours of incubation the total degradability of DM and OM was higher (p < 0.05 for the FM treatment (90.46 and 91.08%, respectively. The nutritional potential of rice bran or parboiled rice bran was confirmed in the diet of ruminants, as it provides a higher fractional rate of in vitro gas production (12.39 and 13.17 mL h-1 and faster degradation of DM and OM in initial times.

  6. N-Terminal-Based Targeted, Inducible Protein Degradation in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Karthik Sekar

    Full Text Available Dynamically altering protein concentration is a central activity in synthetic biology. While many tools are available to modulate protein concentration by altering protein synthesis rate, methods for decreasing protein concentration by inactivation or degradation rate are just being realized. Altering protein synthesis rates can quickly increase the concentration of a protein but not decrease, as residual protein will remain for a while. Inducible, targeted protein degradation is an attractive option and some tools have been introduced for higher organisms and bacteria. Current bacterial tools rely on C-terminal fusions, so we have developed an N-terminal fusion (Ntag strategy to increase the possible proteins that can be targeted. We demonstrate Ntag dependent degradation of mCherry and beta-galactosidase and reconfigure the Ntag system to perform dynamic, exogenously inducible degradation of a targeted protein and complement protein depletion by traditional synthesis repression. Model driven analysis that focused on rates, rather than concentrations, was critical to understanding and engineering the system. We expect this tool and our model to enable inducible protein degradation use particularly in metabolic engineering, biological study of essential proteins, and protein circuits.

  7. Suppression of muscle protein turnover and amino acid degradation by dietary protein deficiency

    Science.gov (United States)

    Tawa, N. E. Jr; Goldberg, A. L.

    1992-01-01

    To define the adaptations that conserve amino acids and muscle protein when dietary protein intake is inadequate, rats (60-70 g final wt) were fed a normal or protein-deficient (PD) diet (18 or 1% lactalbumin), and their muscles were studied in vitro. After 7 days on the PD diet, both protein degradation and synthesis fell 30-40% in skeletal muscles and atria. This fall in proteolysis did not result from reduced amino acid supply to the muscle and preceded any clear decrease in plasma amino acids. Oxidation of branched-chain amino acids, glutamine and alanine synthesis, and uptake of alpha-aminoisobutyrate also fell by 30-50% in muscles and adipose tissue of PD rats. After 1 day on the PD diet, muscle protein synthesis and amino acid uptake decreased by 25-40%, and after 3 days proteolysis and leucine oxidation fell 30-45%. Upon refeeding with the normal diet, protein synthesis also rose more rapidly (+30% by 1 day) than proteolysis, which increased significantly after 3 days (+60%). These different time courses suggest distinct endocrine signals for these responses. The high rate of protein synthesis and low rate of proteolysis during the first 3 days of refeeding a normal diet to PD rats contributes to the rapid weight gain ("catch-up growth") of such animals.

  8. Effects of Two Halophytic Plants (Kochia and Atriplex on Digestibility, Fermentation and Protein Synthesis by Ruminal Microbes Maintained in Continuous Culture

    Directory of Open Access Journals (Sweden)

    A. Riasi

    2012-05-01

    Full Text Available Eight continuous culture fermenters were used in a completely randomized design to evaluate various nutritional values of Kochia (Kochia scoparia compared with Atriplex (Atriplex dimorphostegia. Dried and pelleted samples (leaves and stems provided substrate for metabolism by ruminal microbes maintained in a continuous culture fermentation system. Results indicated that there were no differences (p>0.05 in dry matter (DM and crude protein (CP digestibility between the two halophytic plants. Atriplex had higher (p0.05 between the two halophytic plants in molar proportion of acetate and propionate, but the concentration of butyrate and valerate in Kochia were about two fold of Atriplex (p<0.05. When Kochia provided substrate to the microbes, protein synthesis was higher (p<0.05 compared with feeding Atriplex (5.96 vs. 4.85 g N/kg of OM truly digested. It was concluded that Kochia scoparia and Atriplex dimorphostegia had similar digestibility of DM and CP. It appears that these halophytic plants may not have enough digestible energy for high producing ruminants.

  9. Coordinated Pulses of mRNA and of Protein Translation or Degradation Produce EGF-Induced Protein Bursts.

    Science.gov (United States)

    Golan-Lavi, Roni; Giacomelli, Chiara; Fuks, Garold; Zeisel, Amit; Sonntag, Johanna; Sinha, Sanchari; Köstler, Wolfgang; Wiemann, Stefan; Korf, Ulrike; Yarden, Yosef; Domany, Eytan

    2017-03-28

    Protein responses to extracellular cues are governed by gene transcription, mRNA degradation and translation, and protein degradation. In order to understand how these time-dependent processes cooperate to generate dynamic responses, we analyzed the response of human mammary cells to the epidermal growth factor (EGF). Integrating time-dependent transcript and protein data into a mathematical model, we inferred for several proteins their pre-and post-stimulus translation and degradation coefficients and found that they exhibit complex, time-dependent variation. Specifically, we identified strategies of protein production and degradation acting in concert to generate rapid, transient protein bursts in response to EGF. Remarkably, for some proteins, for which the response necessitates rapidly decreased abundance, cells exhibit a transient increase in the corresponding degradation coefficient. Our model and analysis allow inference of the kinetics of mRNA translation and protein degradation, without perturbing cells, and open a way to understanding the fundamental processes governing time-dependent protein abundance profiles. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

  10. Coordinated Pulses of mRNA and of Protein Translation or Degradation Produce EGF-Induced Protein Bursts

    Directory of Open Access Journals (Sweden)

    Roni Golan-Lavi

    2017-03-01

    Full Text Available Protein responses to extracellular cues are governed by gene transcription, mRNA degradation and translation, and protein degradation. In order to understand how these time-dependent processes cooperate to generate dynamic responses, we analyzed the response of human mammary cells to the epidermal growth factor (EGF. Integrating time-dependent transcript and protein data into a mathematical model, we inferred for several proteins their pre-and post-stimulus translation and degradation coefficients and found that they exhibit complex, time-dependent variation. Specifically, we identified strategies of protein production and degradation acting in concert to generate rapid, transient protein bursts in response to EGF. Remarkably, for some proteins, for which the response necessitates rapidly decreased abundance, cells exhibit a transient increase in the corresponding degradation coefficient. Our model and analysis allow inference of the kinetics of mRNA translation and protein degradation, without perturbing cells, and open a way to understanding the fundamental processes governing time-dependent protein abundance profiles.

  11. Ruminal fermentation and degradation patterns, protozoa population, and urinary purine derivatives excretion in goats and wethers fed diets based on two-stage olive cake: effect of PEG supply.

    Science.gov (United States)

    Yáñez Ruiz, D R; Moumen, A; Martín García, A I; Molina Alcaide, E

    2004-07-01

    Three experiments were conducted in Granadina goats and Segureña wethers fed at maintenance level to evaluate the effect of including a mixture of barley and a new by-product derived from olive oil extraction (two-stage dried olive cake) on ruminal degradation and passage kinetics (Exp. 1), fermentation pattern and protozoa population (Exp. 2), and urinary purine derivatives excretion (Exp. 3). Polyethylene glycol was supplied to the animals to evaluate the effects of tannins contained in the by-product. The experimental diets were as follows: alfalfa hay and alfalfa hay plus a concentrate, formulated with two-stage dried olive cake, barley, and a mineral-vitamin mixture either with or without the addition of polyethylene glycol to the drinking water. The inclusion of two-stage dried olive cake in the diet resulted in an increase of condensed tannins. Ruminal VFA concentration in goats and wethers increased (P two-stage dried olive cake decreased (P fermentation patterns were also similar in goats and wethers and were affected by polyethylene glycol supply. In general, Entodiniomorphida and Holotricha protozoa counts were higher (P two-stage olive cake condensed tannins.

  12. Effect of prolonged intravenous glucose and essential amino acid infusion on nitrogen balance, muscle protein degradation and ubiquitin-conjugating enzyme gene expression in calves

    Directory of Open Access Journals (Sweden)

    Scaife Jes R

    2008-02-01

    Full Text Available Abstract Background Intravenous infusions of glucose and amino acids increase both nitrogen balance and muscle accretion. We hypothesised that co-infusion of glucose (to stimulate insulin and essential amino acids (EAA would act additively to improve nitrogen balance by decreasing muscle protein degradation in association with alterations in muscle expression of components of the ubiquitin-proteasome proteolytic pathway. Methods We examined the effect of a 5 day intravenous infusions of saline, glucose, EAA and glucose + EAA, on urinary nitrogen excretion and muscle protein degradation. We carried out the study in 6 restrained calves since ruminants offer the advantage that muscle protein degradation can be assessed by excretion of 3 methyl-histidine and multiple muscle biopsies can be taken from the same animal. On the final day of infusion blood samples were taken for hormone and metabolite measurement and muscle biopsies for expression of ubiquitin, the 14-kDa E2 ubiquitin conjugating enzyme, and proteasome sub-units C2 and C8. Results On day 5 of glucose infusion, plasma glucose, insulin and IGF-1 concentrations were increased while urea nitrogen excretion and myofibrillar protein degradation was decreased. Co-infusion of glucose + EAA prevented the loss of urinary nitrogen observed with EAA infusions alone and enhanced the increase in plasma IGF-1 concentration but there was no synergistic effect of glucose + EAA on the decrease in myofibrillar protein degradation. Muscle mRNA expression of the ubiquitin conjugating enzyme, 14-kDa E2 and proteasome sub-unit C2 were significantly decreased, after glucose but not amino acid infusions, and there was no further response to the combined infusions of glucose + EAA. Conclusion Prolonged glucose infusion decreases myofibrillar protein degradation, prevents the excretion of infused EAA, and acts additively with EAA to increase plasma IGF-1 and improve net nitrogen balance. There was no evidence of

  13. Influence of barley grain particle size and treatment with citric acid on digestibility, ruminal fermentation and microbial protein synthesis in Holstein calves.

    Science.gov (United States)

    Kazemi-Bonchenari, M; Salem, A Z M; López, S

    2017-08-01

    Chemical and physical treatments of barley grain increase ruminally resistant starch and can improve the rumen fermentation pattern. The objective of the present study was to evaluate the effects of chemical (addition of citric acid, CA) and physical (grinding to two different particle sizes, PS) treatment of barley grain on performance, rumen fermentation, microbial protein yield in the rumen and selected blood metabolites in growing calves. In all, 28 male Holstein calves (172±5.1 kg initial BW) were used in a complete randomised design with a factorial arrangement of 2 barley grain particle sizes×2 levels of citric acid. The diets were as follows: (i) small PS (average 1200 µm) barley grain soaked in water (no CA addition); (ii) small PS barley grain soaked in a CA solution (adding 20 g CA/kg barley); (iii) large PS (average 2400 µm) barley grain soaked in water (no citric acid addition) and (iv) large PS barley grain soaked in a citric acid solution (adding 20 g CA/kg barley). Barley grain was then incorporated at 35% in a total mixed ration and fed to the calves for 11 weeks. Feeding small PS barley decreased feed intake (P=0.02) and average daily weight gain (P=0.01). The addition of CA to barley grain did not affect intake but increased weight gain (P0.05). However, the molar proportion of propionate was increased (P=0.03) when barley was more finely ground, and that of acetate was increased (P=0.04) when CA was added to barley grain. The ruminal concentration of ammonia nitrogen was increased (Pcitric acid increased fibre digestibility of total mixed rations, attenuated the decrease in ruminal pH, and improved weight gain and feed efficiency in male Holstein growing calves fed a high-cereal diet (550 g cereal grain/kg diet).

  14. Abnormal proteins of shortened length are preferentially degraded in the cytosol of cultured MRC5 fibroblasts.

    Science.gov (United States)

    Wharton, S A; Hipkiss, A R

    1984-03-12

    Puromycyl peptides were degraded in MRC5 fibroblasts more rapidly than normal proteins labelled for the corresponding length of time for both long and short labelling periods. The degradation of the puromycyl peptides occurred almost exclusively in the cytosol of the cells. Even when the half-lives of normal and puromycyl peptides were manipulated to be similar, proportionally more of the normal proteins were degraded in the lysosomes. The rapid degradation of the puromycyl peptides was not due to the inhibition of protein synthesis brought about by puromycin but was due to the structure of the substrates themselves. The degree and intracellular site of degradation of puromycyl peptides closely mimic those of abnormal (missense) proteins containing amino acid analogues.

  15. 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

  16. 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.

  17. Protein Degradation Rate in Arabidopsis thaliana Leaf Growth and Development[OPEN

    Science.gov (United States)

    Nelson, Clark J.; Castleden, Ian

    2017-01-01

    We applied 15N labeling approaches to leaves of the Arabidopsis thaliana rosette to characterize their protein degradation rate and understand its determinants. The progressive labeling of new peptides with 15N and measuring the decrease in the abundance of >60,000 existing peptides over time allowed us to define the degradation rate of 1228 proteins in vivo. We show that Arabidopsis protein half-lives vary from several hours to several months based on the exponential constant of the decay rate for each protein. This rate was calculated from the relative isotope abundance of each peptide and the fold change in protein abundance during growth. Protein complex membership and specific protein domains were found to be strong predictors of degradation rate, while N-end amino acid, hydrophobicity, or aggregation propensity of proteins were not. We discovered rapidly degrading subunits in a variety of protein complexes in plastids and identified the set of plant proteins whose degradation rate changed in different leaves of the rosette and correlated with leaf growth rate. From this information, we have calculated the protein turnover energy costs in different leaves and their key determinants within the proteome. PMID:28138016

  18. Casca de algodão em substituição parcial à silagem de capim-elefante para novilhos. 1. Consumo, degradabilidade e digestibilidade total e parcial Partial replacement of elephantgrass silage with cottonseed hulls. 1. Intake, degradability, and apparent ruminal, intestinal, and total tract digestibilities in steers

    Directory of Open Access Journals (Sweden)

    Mario Luiz Chizzotti

    2005-12-01

    randomly assigned to a 4x4 Latin square to study the effects of replacing elephantgrass silage with cottonseed hulls on intake and apparent ruminal, intestinal, and total tract digestibilities of dry matter (DM, organic matter (OM, ether extract (EE, crude protein (CP, neutral detergent fiber corrected to ash and protein (NDFap, and corrected non-fiber carbohydrates (NFCap. Intake of total digestible nutrients (TDN and ruminal degradabilities of cottonseed hulls, elephantgrass silage, sorghum, and soybean meal also were evaluated. Treatments (60% of forage contained on DM basis: 0, 10, 20 or 30% of cottonseed hulls that partially replaced elephantgrass silage in the diet. Digestibilities were obtained using indigestible acid detergent fiber as an internal marker. To determine ruminal degradability of the feeds two steers were used and the following incubation times were adopted: zero, three, six, 12, 24, 48, 72, 96 and 144 hours. Daily intake of all nutrients expressed either as an amount or as percentage of body weight, increased linearly by partially replacing elephantgrass silage with cottonseed hulls. No effects of increasing levels of cottonseed hulls were observed for apparent ruminal, intestinal, and total tract digestibilities of nutrients in the current trial. Consequently, contents of dietary TDN did not differ and averaged 59.53% across diets. In addition, calculated TDN was 55.52% for cottonseed hulls. Effective DM degradation, assuming a passage rate of 5.06%/h, was 38.65; 41.13; 68.58, and 77.56% for cottonseed hulls, elephantgrass silage, sorghum, and soybean meal, respectively. It can be concluded that cottonseed hulls is a good forage alternative to elephantgrass silage and may supply up to 30% of the total DM in diets of dairy steers.

  19. Forage Management Effects on Protein and Fiber Fractions, Protein Degradability, and Dry Matter Yield of Red Clover Conserved as Silage

    Science.gov (United States)

    Due to the action of o-quinones formed via polyphenol oxidase, conserved red clover (Trifolium pratense L.) contains abundant rumen undegradable protein (RUP), but inadequate rumen degradable protein (RDP) for dairy cattle. This study examined how forage management influences RDP, RUP, crude protein...

  20. 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.

  1. Vaccination with recombinant adenovirus expressing peste des petits ruminants virus-F or -H proteins elicits T cell responses to epitopes that arises during PPRV infection.

    Science.gov (United States)

    Rojas, José Manuel; Avia, Miguel; Pascual, Elena; Sevilla, Noemí; Martín, Verónica

    2017-11-21

    Peste des petits ruminants virus (PPRV) causes an economically important disease that limits productivity in small domestic ruminants and often affects the livestock of the poorest populations in developing countries. Animals that survive PPRV develop strong cellular and humoral responses, which are probably necessary for protection. Vaccination should thus aim at mimicking these natural responses. Immunization strategies against this morbillivirus using recombinant adenoviruses expressing PPRV-F or -H proteins can protect PPRV-challenged animals and permit differentiation of infected from vaccinated animals. Little is known of the T cell repertoire these recombinant vaccines induce. In the present work, we identified several CD4+ and CD8+ T cell epitopes in sheep infected with PPRV. We also show that recombinant adenovirus vaccination induced T cell responses to the same epitopes, and led to memory T cell differentiation. T cells primed by these recombinant adenovirus vaccines expanded after PPRV challenge and probably contributed to protection. These data validate the use of recombinant adenovirus expressing PPRV genes as DIVA strategies to control this highly contagious disease.

  2. Teores de proteína bruta para bovinos alimentados com feno de capim-tifton 85: parâmetros ruminais, eficiência de síntese microbiana e degradabilidade in situ Effects of crude protein levels on microbial efficiency and in situ degradability in steers fed Tifton 85 bermudagrass hay supplemented with different protein sources

    Directory of Open Access Journals (Sweden)

    Edilane Aparecida da Silva

    2007-02-01

    Full Text Available Avaliou-se o efeito da deficiência de PB na dieta sobre a eficiência de síntese microbiana e a degradabilidade in situ da MS e FDN em novilhos mestiços. Os tratamentos consistiram de dietas compostas de feno de tifton 85 (FN suplementadas com uréia (FNUR, farelo de soja (FNFS e farelo de glúten de milho- 60 (FNGL. Foram utilizados oito novilhos canulados no rúmen e no duodeno, distribuídos em dois quadrados latinos 4 x 4. O pH e a concentração de nitrogênio não-amoniacal (N-NH3 foram mensurados no fluido ruminal antes e 2, 4, 6 e 8 horas após o fornecimento da ração, utilizando-se a fibra em detergente ácido indigestível (FDAi como indicador do fluxo duodenal. A eficiência microbiana foi determinada por meio das bases purinas. Em todas as dietas, o pH e a concentração de N-NH3 foram adequados para o crescimento dos microrganismos ruminais. Não houve efeito da deficiência de PB na dieta sobre a ingestão de MS, a eficiência de síntese microbiana e a cinética de degradação da MS e FDN do feno de tifton 85. O teor de PB das dietas não alterou o crescimento microbiano, a eficiência de síntese microbiana e a cinética de degradação ruminal.Eight crossbred steers fitted with ruminal and duodenal cannulas were randomly assigned to two replicated 4 x 4 Latin squares to evaluate the effects of crude protein (CP levels on microbial protein synthesis efficiency and in situ degradability of nutrients. Animals received a control diet containing Tifton 85 bermudagrass hay or the control diet supplemented with urea, soybean meal or corn gluten meal-60. Ruminal pH and concentration of ruminal ammonia (NH3-N were determined in the ruminal fluid at 0 (pre-feeding, 2, 4, 6 and 8 hours after feeding. Indigestible acid detergent fiber was used as the internal marker for measuring duodenal flow of nutrients while total purines were used as the direct microbial marker to measure microbial protein synthesis and microbial efficiency

  3. 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.

  4. Targeted Degradation of Proteins Localized in Subcellular Compartments by Hybrid Small Molecules.

    Science.gov (United States)

    Okuhira, Keiichiro; Shoda, Takuji; Omura, Risa; Ohoka, Nobumichi; Hattori, Takayuki; Shibata, Norihito; Demizu, Yosuke; Sugihara, Ryo; Ichino, Asato; Kawahara, Haruka; Itoh, Yukihiro; Ishikawa, Minoru; Hashimoto, Yuichi; Kurihara, Masaaki; Itoh, Susumu; Saito, Hiroyuki; Naito, Mikihiko

    2017-03-01

    Development of novel small molecules that selectively degrade pathogenic proteins would provide an important advance in targeted therapy. Recently, we have devised a series of hybrid small molecules named SNIPER (specific and nongenetic IAP-dependent protein ERaser) that induces the degradation of target proteins via the ubiquitin-proteasome system. To understand the localization of proteins that can be targeted by this protein knockdown technology, we examined whether SNIPER molecules are able to induce degradation of cellular retinoic acid binding protein II (CRABP-II) proteins localized in subcellular compartments of cells. CRABP-II is genetically fused with subcellular localization signals, and they are expressed in the cells. SNIPER(CRABP) with different IAP-ligands, SNIPER(CRABP)-4 with bestatin and SNIPER(CRABP)-11 with MV1 compound, induce the proteasomal degradation of wild-type (WT), cytosolic, nuclear, and membrane-localized CRABP-II proteins, whereas only SNIPER(CRABP)-11 displayed degradation activity toward the mitochondrial CRABP-II protein. The small interfering RNA-mediated silencing of cIAP1 expression attenuated the knockdown activity of SNIPER(CRABP) against WT and cytosolic CRABP-II proteins, indicating that cIAP1 is the E3 ligase responsible for degradation of these proteins. Against membrane-localized CRABP-II protein, cIAP1 is also a primary E3 ligase in the cells, but another E3 ligase distinct from cIAP2 and X-linked inhibitor of apoptosis protein (XIAP) could also be involved in the SNIPER(CRABP)-11-induced degradation. However, for the degradation of nuclear and mitochondrial CRABP-II proteins, E3 ligases other than cIAP1, cIAP2, and XIAP play a role in the SNIPER-mediated protein knockdown. These results indicate that SNIPER can target cytosolic, nuclear, membrane-localized, and mitochondrial proteins for degradation, but the responsible E3 ligase is different, depending on the localization of the target protein. Copyright © 2017 by

  5. Defective folding and rapid degradation of mutant proteins is a common disease mechanism in genetic disorders

    DEFF Research Database (Denmark)

    Gregersen, N; Bross, P; Jørgensen, M M

    2000-01-01

    Many disease-causing point mutations do not seriously compromise synthesis of the affected polypeptide but rather exert their effects by impairing subsequent protein folding or stability of the folded protein. This often results in rapid degradation of the affected protein. The concepts...

  6. Defective folding and rapid degradation of mutant proteins is a common disease mechanism in genetic disorders

    DEFF Research Database (Denmark)

    Gregersen, Niels; Bross, Peter; Jørgensen, Malene Munk

    2000-01-01

    Many disease-causing point mutations do not seriously compromise synthesis of the affected polypeptides but rather exert their effects by impairing subsequent protein folding or stability of the folded protein. This often results in rapid degradation of the affected protein. The concepts...

  7. Forage Polyphenol Oxidase and Ruminant Livestock Nutrition

    Directory of Open Access Journals (Sweden)

    Michael Richard F. Lee

    2014-12-01

    Full Text Available Polyphenol oxidase (PPO is associated with the detrimental effect of browning fruit and vegetables, however interest within PPO containing forage crops has grown since the brownng reaction was associated with reduced nitrogen (N losses in silo and the rumen. The reduction in protein breakdown in silo of red clover (high PPO forage increased the quality of protein, improving N-use efficiency (NUE when fed to ruminants. A further benefit of red clover silage feeding is a significant reduction in lipolysis in silo and an increase in the deposition of beneficial C18 polyunsaturated fatty acid (PUFA in animal products, which has also been linked to PPO activity. PPOs protection of plant protein and glycerol based-PUFA in silo is related to the deactivation of plant proteases and lipases. This deactivation occurs through PPO catalysing the conversion of diphenols to quinones which bind with cellular nucleophiles such as protein reforming a protein-bound phenol (PBP. If the protein is an enzyme the complexing denatures the enzyme. However, PPO is inactive in the anaerobic rumen and therefore any subsequent protection of plant protein and glycerol based-PUFA in the rumen must be as a result of events that occurred to the forage pre-ingestion. Reduced activity of plant proteases and lipases would have little effect on NUE and glycerol based-PUFA in the rumen due to the greater concentration of rumen microbial proteases and lipases. The mechanism for PPOs protection of plant protein in the rumen is a consequence of complexing plant protein, rather than protease deactivation per se. These complexed proteins reduce protein digestibility in the rumen and subsequently increase un-degraded dietary protein flow to the small intestine. The mechanism for protecting glycerol-based PUFA has yet to be fully elucidated but may be associated with entrapment within PBP reducing access to microbial lipases or differences in rumen digestion kinetics of red clover.

  8. Signal-transduction networks and the regulation of muscle protein degradation.

    Science.gov (United States)

    Szewczyk, Nathaniel J; Jacobson, Lewis A

    2005-10-01

    Protein degradation in muscle functions in maintaining normal physiological homeostasis and adapting to new homeostatic states, and is required for muscle wasting or atrophy in various pathological states. The interplay between protein synthesis and degradation to maintain homeostasis is complex and responds to a variety of autocrine and intercellular signals from neuronal inputs, hormones, cytokines, growth factors and other regulatory molecules. The intracellular events that connect extracellular signals to the molecular control of protein degradation are incompletely understood, but likely involve interacting signal-transduction networks rather than isolated pathways. We review some examples of signal-transduction systems that regulate protein degradation, including effectors of proteolysis inducing factor (PIF), insulin and insulin-like growth factor (IGF) and their receptors, and fibroblast growth factor (FGF) and its receptors.

  9. Wet degradation of keratin proteins : Linking amino acid, elemental and isotopic composition

    NARCIS (Netherlands)

    Von Holstein, I. C C; Penkman, Kirsty E H; Peacock, E.E.; Collins, M. J.

    2014-01-01

    Rationale Archaeological keratin samples are increasingly the subject of palaeodietary, provenancing and dating studies. Keratin samples from wet archaeological contexts are microbiologically and chemically degraded, causing differential diagenesis of protein structures in hair fibres. The effects

  10. Anaerobic degradation of cellulosic substrates - Bionic implementation of the forestomach sysem of a ruminant; Anaerober Abbau cellulosehaltiger Substrate. Bionische Implementierung des Vormagensystems des Wiederkaeuers

    Energy Technology Data Exchange (ETDEWEB)

    Weichgrebe, Dirk [Hannover Univ. (Germany). Inst. fuer Siedlungswasserwirtschaft und Abfalltechnik; Stopp, Paul; Rosenwinkel, Karl-Heinz; Breves, Gerhard; Strecker, Michael

    2011-07-01

    The forestomach system of a ruminant technically can be realized by means of a two-stage arrangement of hydrolysis/acidification (first step) and methanation (second stage). Both stages are connected by a retention of solid substances and by a recirculation of process water. The first stage converts cellulose-rich substrates in short-chain volatile organic acids. The second stage converts these short-chain volatile organic acids into biogas. The technical realization of this two-stage arrangement is investigated by the co-operation project RUMEN-DAUMEN 2.0 of the Institute of Environmental Engineering at the Leibniz University of Hannover (Hannover, Federal Republic of Germany) and the Physiological Institute of the University of Veterinary Medicine Hannover (Hannover, Federal Republic of Germany). The aim of this study is to design a viable implementation of cellulosic waste materials by means of an efficient hydrolysis and to convert produced fatty acids efficiently in biogas.

  11. Monitoring of the Enzymatic Degradation of Protein Corona and Evaluating the Accompanying Cytotoxicity of Nanoparticles.

    Science.gov (United States)

    Ma, Zhifang; Bai, Jing; Jiang, Xiue

    2015-08-19

    Established nanobio interactions face the challenge that the formation of nanoparticle-protein corona complexes shields the inherent properties of the nanoparticles and alters the manner of the interactions between nanoparticles and biological systems. Therefore, many studies have focused on protein corona-mediated nanoparticle binding, internalization, and intracellular transportation. However, there are a few studies to pay attention to if the corona encounters degradation after internalization and how the degradation of the protein corona affects cytotoxicity. To fill this gap, we prepared three types of off/on complexes based on gold nanoparticles (Au NPs) and dye-labeled serum proteins and studied the extracellular and intracellular proteolytic processes of protein coronas as well as their accompanying effects on cytotoxicity through multiple evaluation mechanisms, including cell viability, adenosine triphosphate (ATP) content, mitochondrial membrane potential (MMP), and reactive oxygen species (ROS). The proteolytic process was confirmed by recovery of the fluorescence of the dye-labeled protein molecules that was initially quenched by Au NPs. Our results indicate that the degradation rate of protein corona is dependent on the type of the protein based on systematical evaluation of the extracellular and intracellular degradation processes of the protein coronas formed by human serum albumin (HSA), γ-globulin (HGG), and serum fibrinogen (HSF). Degradation is the fastest for HSA corona and the slowest for HSF corona. Notably, we also find that the Au NP-HSA corona complex induces lower cell viability, slower ATP production, lower MMP, and higher ROS levels. The cytotoxicity of the nanoparticle-protein corona complex may be associated with the protein corona degradation process. All of these results will enrich the database of cytotoxicity induced by nanomaterial-protein corona complexes.

  12. Fermentação e Degradabilidade Ruminal em Bovinos Alimentados com Resíduos de Mandioca e Cana-de-Açúcar ensilados com Polpa Cítrica Peletizada Ruminal Fermentation and Degradability in Bovine Fed Diet with Cassava Residue and Sugar Cane Ensiled with Pelleted Citrus Pulp

    Directory of Open Access Journals (Sweden)

    Roselene Nunes da Silveira

    2002-01-01

    Full Text Available O presente trabalho foi conduzido com o objetivo de avaliar a degradabilidade ruminal da matéria seca (MS, fibra em detergente neutro (FDN e amido, além de pH, amônia e ácidos graxos voláteis ruminais, em bovinos alimentados com silagens de milho (SMi, de raspa de mandioca com polpa cítrica (SRp, de casca de mandioca com polpa cítrica (SCc e de cana-de-açúcar com polpa cítrica (SCn. Foram utilizados quatro novilhos, mestiços, castrados, canulados no rúmen e duodeno, em quatro períodos experimentais, com 11 dias de adaptação à dieta e oito dias de coleta. O delineamento experimental foi o quadrado latino 4x4. Foram adotados oito horários para a incubação das silagens: 3, 6, 12, 24, 48, 72, 96 e 120 horas. A SRp apresentou maior degradação efetiva (Kp 5% da MS e da FDN (48,44 e 45,78%, respectivamente, quando comparada com a SMi (45,50 e 23,75%, a SCc (43,87 e 24,20% e a SCn (40,76 e 25,78%. Para todos os tratamentos, o pH e a concentração de N-NH3 ruminal foram adequados para o crescimento dos microrganismos ruminais. Os valores de AGV para os tratamentos de SMi, SRp e SCc foram semelhantes entre si e superiores aos do tratamento com SCn.The objective of this work was to evaluate the dry matter (DM, neutral detergente fiber (NDF and starch ruminal degradability besides pH, N-NH3 and valatile fatty acids (VFA in bovines fed diet with corn (CS, cassava meal (CMS, cassava hull (CHS, and sugar cane (SCS. The CMS, CHS and SCS were ensiled with citrus pulp (CPP. Four crossbred, castrated, rumen and duodenum cannulated steers were used in four experimental periods, with 11 days for diet adaptation and 8 days for sampling. The experimental design was 4x4 Latin Square. There were eight incubations times: 3, 6, 12, 24, 48, 72, 96 and 120 hours. The CMS showed higher DM, and NDF effective degradation (Kp 5% (48.44 and 45.78%, respectively than CS (45.50, 23.75%, CHS (43.87, 24.20% and SCS (40.76, 25.78%. For all the treatments, the p

  13. Stromal protein degradation is incomplete in Arabidopsis thaliana autophagy mutants undergoing natural senescence

    Directory of Open Access Journals (Sweden)

    Lee Travis A

    2013-01-01

    Full Text Available Abstract Background Degradation of highly abundant stromal proteins plays an important role in the nitrogen economy of the plant during senescence. Lines of evidence supporting proteolysis within the chloroplast and outside the chloroplast have been reported. Two extra-plastidic degradation pathways, chlorophagy and Rubisco Containing Bodies, rely on cytoplasmic autophagy. Results In this work, levels of three stromal proteins (Rubisco large subunit, chloroplast glutamine synthetase and Rubisco activase and one thylakoid protein (the major light harvesting complex protein of photosystem II were measured during natural senescence in WT and in two autophagy T-DNA insertion mutants (atg5 and atg7. Thylakoid-localized protein decreased similarly in all genotypes, but stromal protein degradation was incomplete in the two atg mutants. In addition, degradation of two stromal proteins was observed in chloroplasts isolated from mid-senescence leaves. Conclusions These data suggest that autophagy does contribute to the complete proteolysis of stromal proteins, but does not play a major degenerative role. In addition, support for in organello degradation is provided.

  14. 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.

  15. 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.

  16. Degradação ruminal da fração fibra de silagens de milho sem espigas com cana-de-açúcar e bagaço de mandioca = Fiber fraction ruminal degradation of corn silage without spikes with sugar cane and manioc pulp

    Directory of Open Access Journals (Sweden)

    Marcos Elias Traad da Silva

    2006-10-01

    Full Text Available Avaliou-se a degradação ruminal da fibra em detergente neutro (FDN e em detergente ácido (FDA de silagens de milho sem espigas (SMSE com cana -de-açúcar (CA e bagaço seco de mandioca (BSM, comparadas com a silagem de milho com espigas (SMCE. Os alimentos foram ensilados com (CI e sem inoculante (SI - Lactobacillus plantarum e L. paracasei ssp. paracasei. Estudaram-se: as frações solúvel (A, potencialmente degradável (B e não-degradável (C; a taxa de degradação da fração B (c; a degradabilidade efetiva (DE e potencial (DP. Os tratamentos foram: T1 (SMSE + 20% CA-SI; T2 (SMSE + 20% CA-CI; T3 (SMSE + 35% CA-SI; T4 (SMSE + 35% CA-CI; T5 (SMSE+ 20% BSM-SI; T6 (SMSE + 20% BSM-CI; T7 (SMSE + 35% BSM-SI; T8 (SMSE + 35% BSM-CI; T9 (SMCE-SI e T10 (SMCE-CI. Médias foram comparadas em um delineamento inteiramente casualisado. As mai ores (p0,05 para a fração C da FDA dos tratamentos T3 (C = 20,7%, T4 (C = 22,6%, T9 (C = 22,7% e T10 (C = 24,0%. Os resultados indicam que há possibilidade para a inclusão do BSM no lugar das espigas de milho verde para a produção de silagem até o nível de 35%.This study aimed to evaluate the ruminal degradation of corn silages without spikes (CSWS, with addit ion of sugar cane (SC and dry manioc pulp (DMW, compared to corn silage (CS. The feeds were ensiled with (WI and without (IF inoculants - Lactobacillus plantarum and L. paracasei ssp. paracasei. The following fractionswere studied: water soluble (A, potentially degradable (B, non degradable (C, fraction B degradable ratio (c; effective (ED and potential (PD degradability. The treatments evaluated were: T1 (CSWS + 20% SC -IF; T2 (CSWS + 20% SC-WI; T3 (CSWS + 35% SC-IF; T4 (CSWS + 35% SC -WI; T5 (CSWS + 20% DMW-IF; T6 (CSWS + 20% DMW-WI; T7 (CSWS + 35% DMW -IF; T8 (CSWS + 35% DMW-WI; T9 (CS-WI and T10 (CS-IF. The average differences were compared on a completely randomized model. Higher (p0.05 were observed between ADF C fraction for T3 (C=20.7%, T4

  17. Investigating Molecular Structures of Bio-Fuel and Bio-Oil Seeds as Predictors To Estimate Protein Bioavailability for Ruminants by Advanced Nondestructive Vibrational Molecular Spectroscopy.

    Science.gov (United States)

    Ban, Yajing; L Prates, Luciana; Yu, Peiqiang

    2017-10-18

    This study was conducted to (1) determine protein and carbohydrate molecular structure profiles and (2) quantify the relationship between structural features and protein bioavailability of newly developed carinata and canola seeds for dairy cows by using Fourier transform infrared molecular spectroscopy. Results showed similarity in protein structural makeup within the entire protein structural region between carinata and canola seeds. The highest area ratios related to structural CHO, total CHO, and cellulosic compounds were obtained for carinata seeds. Carinata and canola seeds showed similar carbohydrate and protein molecular structures by multivariate analyses. Carbohydrate molecular structure profiles were highly correlated to protein rumen degradation and intestinal digestion characteristics. In conclusion, the molecular spectroscopy can detect inherent structural characteristics in carinata and canola seeds in which carbohydrate-relative structural features are related to protein metabolism and utilization. Protein and carbohydrate spectral profiles could be used as predictors of rumen protein bioavailability in cows.

  18. 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

  19. Protein degradation corrects for imbalanced subunit stoichiometry in OST complex assembly.

    Science.gov (United States)

    Mueller, Susanne; Wahlander, Asa; Selevsek, Nathalie; Otto, Claudia; Ngwa, Elsy Mankah; Poljak, Kristina; Frey, Alexander D; Aebi, Markus; Gauss, Robert

    2015-07-15

    Protein degradation is essential for cellular homeostasis. We developed a sensitive approach to examining protein degradation rates in Saccharomyces cerevisiae by coupling a SILAC approach to selected reaction monitoring (SRM) mass spectrometry. Combined with genetic tools, this analysis made it possible to study the assembly of the oligosaccharyl transferase complex. The ER-associated degradation machinery compensated for disturbed homeostasis of complex components by degradation of subunits in excess. On a larger scale, protein degradation in the ER was found to be a minor factor in the regulation of protein homeostasis in exponentially growing cells, but ERAD became relevant when the gene dosage was affected, as demonstrated in heterozygous diploid cells. Hence the alleviation of fitness defects due to abnormal gene copy numbers might be an important function of protein degradation. © 2015 Mueller 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).

  20. 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.

  1. 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…

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

    African Journals Online (AJOL)

    Diets were formulated to be iso-nitrogenous and iso-caloric, and differed in undegradable dietary protein content. In Experiment 1 calves were randomly assigned to one of three dietary treatments containing low (LD), medium (MD) or high (HD) levels of rumen degradable protein. In Experiment 2 calves received a starter ...

  3. Degradation of HaloTag-fused nuclear proteins using bestatin-HaloTag ligand hybrid molecules.

    Science.gov (United States)

    Tomoshige, Shusuke; Naito, Mikihiko; Hashimoto, Yuichi; Ishikawa, Minoru

    2015-10-14

    We have developed a protein knockdown technology using hybrid small molecules designed as conjugates of a ligand for the target protein and a ligand for ubiquitin ligase cellular inhibitor of apoptosis protein 1 (cIAP1). However, this technology has several limitations. Here, we report the development of a novel protein knockdown system to address these limitations. In this system, target proteins are fused with HaloTag to provide a common binding site for a degradation inducer. We designed and synthesized small molecules consisting of alkyl chloride as the HaloTag-binding degradation inducer, which binds to HaloTag, linked to BE04 (2), which binds to cIAP1. Using this system, we successfully knocked down HaloTag-fused cAMP responsive element binding protein 1 (HaloTag-CREB1) and HaloTag-fused c-jun (HaloTag-c-jun), which are ligand-unknown nuclear proteins, in living cells. HaloTag-binding degradation inducers can be synthesized easily, and are expected to be useful as biological tools for pan-degradation of HaloTag-fused proteins.

  4. 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…

  5. Chemical composition and ruminal degradability of the sugar cane silage treated with chemical and bacterial additivesComposição química e degradabilidade ruminal de silagens da cana-de-açúcar tratada com aditivos químicos e bacteriano

    Directory of Open Access Journals (Sweden)

    Luiz Eduardo dos Santos

    2012-02-01

    Full Text Available The objective of this trial was to evaluate the effect of the addition of chemical and bacterial additive in the ensiling of sugar cane (Saccharum officinarum L. on chemical composition, pH, kinectic fraction and in situ degradation of nutritions components of silages. Five rumen-cannulated ½ Simental + ½ Zebu steers were allotted to a completely randomized design. The steers were placed in individual cages and they were fed with diets with 76% forage (%DM. Five silages were evaluated: control – sugar cane, no additives; urea – sugar cane + 0.5% of urea (wet basis; inoculant – sugar cane inoculated with LactoSilo® (390 g/40 t forage; NaOH – sugar cane + 1.0% of sodium hydroxide (wet basis; CaOH – sugar cane + 0.6% of calcium hydroxide (wet basis. The silage additives with sodium hydroxide showed the highest pH values before (11.20 and after (4.87 for silage. No differences were observed among the silages for dry matter (26.85, crude protein (5.25 and acid detergent fiber (57.21. Fractionation of dry matter and organic matter of silages showed similar behavior, with higher values of the soluble fraction (fraction A for silages with sodium hydroxide (45.86 and 30.95% and calcium hydroxide (29.47 and 26.13%. The use of sodium hydroxide allowed obtaining higher values for the degradation of cell wall components of silages from cane sugar. The potencial and effective degradability with 3, 5 and 8%/h of passage rate were respectively 88.44, 64.45, 56.73 and 49.83% for NDF and 82.57, 55.51, 46.72 and 38.83% for ADF, indicating that the use of sodium hydroxide as chemical additives can improve the nutritive value of cane sugar silage.Objetivou-se avaliar o efeito da inclusão de aditivos químicos e bacterianos na ensilagem de cana-de-açúcar sobre a composição, pH, fracionamento e cinética de degradação in situ dos componentes nutritivos. Utilizaram-se cinco bovinos ½ Simental + ½ Zebu providos de cânula ruminal, alocados em

  6. Cinética da degradação ruminal dos carboidratos de quatro gramíneas tropicais em diferentes idades de corte e doses de adubação nitrogenada: técnica de produção de gases Kinetic parameters of carbohydrates ruminal degradation of four tropical grasses in different cutting ages and nitrogen fertilizer levels

    Directory of Open Access Journals (Sweden)

    Edenio Detmann

    2009-01-01

    Full Text Available Objetivou-se neste experimento avaliar a cinética de degradação ruminal dos capins setária (Setaria anceps Stapf, hemarthria (Hemarthria altissima [Poir] Stapf. & Hubbard, angola (Brachiaria purpurascens [Raddi] Henr. e acroceres (Acroceras macrum Stapf. adubados com 0, 100, 200, 300 ou 400 kg de N/ha e colhidos aos 28, 42, 56 ou 70 dias de idade. Os resultados foram avaliados por intermédio de análise de fatores. Após redução e avaliação da variação conjunta total das variáveis, optou-se pela adoção de três fatores, que englobaram 86,4% da variação total: o primeiro fator (Fator 1 associou-se ao volume de gás oriundo das frações de lenta e de rápida degradação e à taxa de degradação da fração rapidamente degradável (Vf1, Vf2 e k2; o segundo fator (Fator 2 associou-se à taxa de degradação de lenta digestão e à latência (k1 e L; e o terceiro fator (Fator 3, por sua vez, associou-se à digestibilidade in vitro da matéria seca (DIVMS. Os parâmetros da cinética de degradação foram influenciados pelas idades de corte. A digestibilidade in vitro da matéria seca reduziu com o avançar da idade das plantas. A adubação nitrogenada não promoveu respostas evidentes nos parâmetros avaliados. O capim-hemarthria se destacou dos demais em virtude de maior digestibilidade.The objective of this experiment was to evaluate the effects of levels 0, 100, 200, 300 and 400 kg of nitrogen/ha and cutting ages of 28, 42, 56 and 70 days on the kinetic parameters of ruminal degradation of carbohydrate of the following tropical forages: Setaria grass (Setaria anceps Stapf, Limpo grass (Hemarthria altissima [Poir] Stapf. & Hubbard, California grass (Brachiaria purpurascens [Raddi] Henr. and Nilo grass (Acroceras macrum Stpaf. The results were submitted to a factor analysis. After reduction and evaluation of the combined variation from the total variables, three factors comprising 86.44% of the total variation were considered

  7. Oxidized mitochondrial protein degradation and repair in aging and oxidative stress.

    Science.gov (United States)

    Ugarte, Nicolas; Petropoulos, Isabelle; Friguet, Bertrand

    2010-08-15

    Proteins are main targets for oxidative damage that occurs during aging and in oxidative stress situations. Since the mitochondria is a major source of reactive oxygen species, mitochondrial proteins are especially exposed to oxidative modification, and elimination of oxidized proteins is crucial for maintaining the integrity of this organelle. Hence, enzymatic reversal of protein oxidation and protein degradation is critical for protein homeostasis while protein maintenance failure has been implicated in the age-related accumulation of oxidized proteins. Within the mitochondrial matrix, the ATP-stimulated mitochondrial Lon protease is believed to play an important role in the degradation of oxidized protein, and age-associated impairment of Lon-like protease activity has been suggested to contribute to oxidized protein buildup in the mitochondria. Oxidized protein repair is limited to certain oxidation products of the sulfur-containing amino acids cysteine and methionine. Oxidized protein repair systems, thioredoxin/thioredoxin reductase or glutaredoxin/glutathione/glutathione reductase that catalytically reduce disulfide bridges or sulfenic acids, and methionine sulfoxide reductase that reverses methionine sulfoxide back to methionine within proteins, are present in the mitochondrial matrix. Thus, the role of the mitochondrial Lon protease and the oxidized protein repair system methionine sulfoxide reductase is further addressed in the context of oxidative stress and aging.

  8. Cinética ruminal do feno de Stylosanthes guianensis Ruminal kinetics of Stylosanthes guianensis hay

    Directory of Open Access Journals (Sweden)

    M.M. Ladeira

    2001-04-01

    .1% and 30.3%, respectively. The degradation rate of crude protein (CP was 9.7%/h, the potential degradability was 56.0% and the effective degradabilty was 47.5%. The cellulose presented larger effective degradability than hemicellulose, with the respective values of 22.5% and 8.9%. The passage rate for solids was 2.7%/h. The pH decreased linearly with time of rumen fluid sampling. For the time of 5.13 hours after feeding, the highest ammonia concentration (12.18 mg/100ml was observed. The S. guinanensis hay showed high degradation rates and low ruminal degradability of DM and CP.

  9. 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 ...

  10. The effect of different rumen degradable protein concentrations and ...

    African Journals Online (AJOL)

    Nolte Joubert

    that tryptophan synthesis might be dependent on peptides and AA's. The initial decrease in the ratios of essential AA's to lysine between the control treatment and 40 g supplemental RDP/d probably resulted from enhanced microbial growth rates (Argyle & Baldwin, 1989; Nolte et al., 2003) that lead to reduced protein.

  11. Small-molecule hydrophobic tagging-induced degradation of HaloTag fusion proteins.

    Science.gov (United States)

    Neklesa, Taavi K; Tae, Hyun Seop; Schneekloth, Ashley R; Stulberg, Michael J; Corson, Timothy W; Sundberg, Thomas B; Raina, Kanak; Holley, Scott A; Crews, Craig M

    2011-07-03

    The ability to regulate any protein of interest in living systems with small molecules remains a challenge. We hypothesized that appending a hydrophobic moiety to the surface of a protein would mimic the partially denatured state of the protein, thus engaging the cellular quality control machinery to induce its proteasomal degradation. We designed and synthesized bifunctional small molecules to bind a bacterial dehalogenase (the HaloTag protein) and present a hydrophobic group on its surface. Hydrophobic tagging of the HaloTag protein with an adamantyl moiety induced the degradation of cytosolic, isoprenylated and transmembrane HaloTag fusion proteins in cell culture. We demonstrated the in vivo utility of hydrophobic tagging by degrading proteins expressed in zebrafish embryos and by inhibiting Hras1(G12V)-driven tumor progression in mice. Therefore, hydrophobic tagging of HaloTag fusion proteins affords small-molecule control over any protein of interest, making it an ideal system for validating potential drug targets in disease models.

  12. Small-Molecule Hydrophobic Tagging Induced Degradation of HaloTag Fusion Proteins

    Science.gov (United States)

    Neklesa, Taavi K.; Tae, Hyun Seop; Schneekloth, Ashley R.; Stulberg, Michael J.; Corson, Timothy W.; Sundberg, Thomas B.; Raina, Kanak; Holley, Scott A.; Crews, Craig M.

    2011-01-01

    The ability to regulate any protein of interest in living systems with small molecules remains a challenge. We hypothesized that appending a hydrophobic moiety to the surface of a protein would mimic the partially denatured state of the protein, thus engaging the cellular quality control machinery to induce its proteasomal degradation. We designed and synthesized bifunctional small molecules that bind a bacterial dehalogenase (HaloTag protein) and present a hydrophobic group on its surface. Remarkably, hydrophobic tagging of the HaloTag protein with an adamantyl moiety induced the degradation of cytosolic, isoprenylated, and transmembrane fusion proteins in cell culture. We demonstrated the in vivo utility of hydrophobic tagging by degrading proteins expressed in zebrafish embryos and by inhibiting RasG12V-driven tumor progression in mice. Therefore, hydrophobic tagging of HaloTag fusion proteins affords small molecule control over any protein of interest, making it an ideal system for validating potential drug targets in disease models. PMID:21725302

  13. Characterization of N-Acetyl-Tryptophan Degradation in Protein Therapeutic Formulations.

    Science.gov (United States)

    Hogan, Kyle L; Leiske, Danielle; Salisbury, Cleo M

    2017-12-01

    N-Acetyl-tryptophan (NAT) is used as a stabilizer for preparations of human serum albumin and has more recently been demonstrated to provide oxidative protection for labile Trp residues in monoclonal antibodies. As a component in the formulations of protein therapeutics, NAT is sacrificially degraded; therefore, understanding the identity and quantity of NAT degradants potentially formed in these drug products is essential to understanding the potential patient impact of this additive. Here, we report a simple reversed-phase chromatography approach that allows systematic investigation of NAT degradation in relevant formulations under stressed conditions. Screening a panel of NAT-containing samples following a variety of forced stress conditions led to a range of NAT degradation from minimal (3%) to significant (83%). NAT degradants were observed to be largely conserved between oxidative and thermal stress conditions. Online mass spectrometry and standard compound synthesis allowed for identification of the major degradants in the stressed sample panel. NAT degradation was minimal under recommended storage conditions and in relevant thermal stress conditions for a representative protein therapeutic drug product, suggesting that NAT is stable under normal manufacturing, storage, and handling conditions. This work supports the use of NAT as an antioxidant in liquid drug product formulations. Copyright © 2017 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

  14. 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

    Full Text Available Objetivou-se avaliar o efeito de diferentes relações entre proteína verdadeira e nitrogênio não-proteico sobre o consumo e a dinâmica de trânsito e degradação ruminal da fibra em detergente neutro (FDN da forragem em bovinos em pastejo durante o período das águas. Foram utilizados cinco novilhos mestiços Holandês × Zebu, castrados, com peso corporal inicial de 335±35 kg fistulados no rúmen e no abomaso. Os tratamentos foram: controle (somente pasto; e suplementos com ureia; com 2/3 de compostos nitrogenados oriundos da ureia e 1/3 de compostos nitrogenados oriundos da albumina; com 1/3 de compostos nitrogenados oriundos da ureia e 2/3 de compostos nitrogenados oriundos da albumina; e com albumina. Foram fornecidos 200 g/dia de proteína bruta (PB a partir dos suplementos. O experimento foi conduzido segundo delineamento em quadrado latino 5 × 5, com cinco períodos experimentais de 15 dias. Não foram observados efeitos da suplementação sobre o consumo voluntário, com exceção do consumo de PB, que aumentou com a suplementação. A substituição da ureia por albumina nos suplementos teve efeito linear sobre o consumo de PB. Os consumos dos demais componentes da dieta não foram afetados pela composição dos suplementos. Nenhum efeito foi observado sobre a taxa de passagem ruminal de compostos fibrosos. O fornecimento de suplementos ampliou, em média, a estimativa da taxa comum de latência e degradação da FDN. Contudo, não houve efeito da alteração na composição dos suplementos sobre este parâmetro. A suplementação de bovinos com fontes de compostos nitrogenados degradáveis no rúmen proteicos ou não-proteicos durante o período das águas não afeta o consumo voluntário de pasto.The objective of this study was to evaluate the effect of different true protein:non-protein nitrogen ratios in supplements on intake and ruminal transit and degradation dynamics of neutral detergent fiber (NDF in grazing cattle

  15. Active Degradation Explains the Distribution of Nuclear Proteins during Cellular Senescence.

    Directory of Open Access Journals (Sweden)

    Enrico Giampieri

    Full Text Available The amount of cellular proteins is a crucial parameter that is known to vary between cells as a function of the replicative passages, and can be important during physiological aging. The process of protein degradation is known to be performed by a series of enzymatic reactions, ranging from an initial step of protein ubiquitination to their final fragmentation by the proteasome. In this paper we propose a stochastic dynamical model of nuclear proteins concentration resulting from a balance between a constant production of proteins and their degradation by a cooperative enzymatic reaction. The predictions of this model are compared with experimental data obtained by fluorescence measurements of the amount of nuclear proteins in murine tail fibroblast (MTF undergoing cellular senescence. Our model provides a three-parameter stationary distribution that is in good agreement with the experimental data even during the transition to the senescent state, where the nuclear protein concentration changes abruptly. The estimation of three parameters (cooperativity, saturation threshold, and maximal velocity of the reaction, and their evolution during replicative passages shows that only the maximal velocity varies significantly. Based on our modeling we speculate the reduction of functionality of the protein degradation mechanism as a possible competitive inhibition of the proteasome.

  16. Optimum conditions for enzymatic degradation of some oilseed proteins

    Directory of Open Access Journals (Sweden)

    El-Zanaty, E. A.

    2002-09-01

    Full Text Available Soybean, sesame seed, and rice bran meal proteins were hydrolyzed with two enzymes, namely, papain and bromelain. Experiments were carried out to elucidate the optimum condition for each enzyme when acting on each substrate seperately. Results revealed that the highest relative activities for papain were achieved with E/S 0.06 , 0.29, 0.19 and pH 7.2, 7.0, 7.0 for soybean, sesame,and rice bran meal proteins, respectively. Optimum temperature for papain while hydrolysing the three substrates was 50 ºC. When using bromelain optimum E/S resulting in highest relative activities were 0.067, 0.058 and 0.21 for soybean, sesame,and rice bran meal protein, respectively. Optimum pH was 6.0 and optimum temperature was 45 ºC for bromelain when hydrolysing the protein of the three substrates. A numerical correlation of enzymatic behaviour for the different substrates was calculated.Proteínas de haba de soja, semilla de sésamo y harina de germen de arroz se hidrolizaron con dos enzimas, denominadas, papaina y bromelaina. Se han llevado a cabo experimentos para determinar las condiciones óptimas de cada enzima cuando actúan separadamente sobre cada sustrato. Los resultados mostraron que las mayores actividades relativas para la papaina se consiguieron con una E/S 0,06, 0,29, 0,19 y un pH 7.2, 7.0, 7.0 para las proteínas de haba de soja, sésamo y harina de germen de arroz, respectivamente. La temperatura óptima para la papaina durante la hidrólisis de los tres sustratos fue de 50 ºC. Cuando se usa bromelaina las relaciones E/S óptimas que proporcionaron mayor actividad relativa fueron 0.067, 0.058 y 0.21 para las proteínas de habas de soja, sésamo y harina de germen de arroz respectivamente. El pH óptimo fue 6.0 y la temperatura óptima 45 ºC para la bromelaina cuando se hidroliza la proteína de los tres sustratos. Con estos datos se hizo una correlación numérica del comportamiento enzimático para los diferentes sustratos.

  17. Avaliação da Composição de Vários Alimentos e Determinação da Cinética Ruminal da Proteína, Utilizando o Método de Produção de Gás e Amônia in Vitro Chemical Composition Evaluation and Ruminal Protein Kinetics of Some Feedstuffs Using a Gas and Ammonia Production in vitro Method

    Directory of Open Access Journals (Sweden)

    Fernando Iván Londoño Hernández

    2002-02-01

    nitrogen compounds of 24 concentrate feedstuffs and 10 grasses were made using the concentrations of soluble nitrogen in TCA and gas production. It was used 200 mL of ruminal fluid and 800 mL of medium for 100 vessels. It was used 1.0 g of trypticase and 0.1 mL of microminerals solution to prepare 400 mL of medium. It was used 0.8 g of ammonium bicarbonate and 7 g of sodium bicarbonate to prepare 200 mL of buffer solution and, to prepare 200 mL of macromineral solution, 1.15 g of Na2HPO4 anhydrous, 1.25 g of KH2PO4 anhydrous and 0.1 g of MgSO4.7H2O were weighed. Reducing solution was prepared with 0.64 g of cysteine-HCL, 0.64 g of sodium sulfide and 4 mL of 1N NaOH. The disappearance of nitrogen compounds of feedstuffs was determined at 6 and 12 hours, where 1.875 mg of N was incubated with 0, 33, 67, 100 mg oh starch, 6 mL of medium, 4 mL of ruminal fluid-medium mixture and 0.1 mL of reducing solution using CO2. Data of degradation rates indicated that corn gluten feed, casein, dry grounded peanut grain, cassava rasp, sorghum silage with or without inoculum, corn silage and honeygrass showed the highest rates of protein degradation and the slowest degradation rates were obtained by sugar cane yeast, feather meal, fish meal, broiler litter using wood rind as adsorvent and signalgrass. Estimates of degradation rates of forage feedstuffs were higher than degradation rates estimated previously by an inhibitor method. It is recommended to use 12 hours for incubation of concentrate feedstuffs and 6 hours for grasses.

  18. An assessment of the ability of the stay-green phenotype in lolium species to provide an improved protein supply for ruminants.

    Science.gov (United States)

    Kingston-Smith, Alison H; Bollard, Andrea L; Humphreys, Mervyn O; Theodorou, Michael K

    2002-06-01

    The stay-green phenotype results from a naturally occurring mutation in which senescent leaves retain their chlorophyll and the associated apoprotein, LHCPII. Protection of this protein pool could deliver grass with enhanced protein content and could decrease the extent of protein degradation by plant proteases in the rumen. This would enhance the efficiency of protein utilization in livestock to the benefit of the environment. Field plots of stay-green and wild-type Lolium perenne were defoliated at intervals to simulate grazing. There were variations in foliar protein content and proteolysis throughout the year, but no significant differences between genotypes when material was analysed fresh or after it was cut and dried to simulate hay-making, which possibly induced senescence. In a subsequent experiment with stay-green and wild-type L temulentum, increased protein retention and decreased protein degradability were observed in stay-green leaves that were allowed to senescence naturally and extensively on the plant. That there is no difference between the two L. perenne genotypes suggests that as a field crop in grazed pastures the stay-green genotype would not confer a nutritional advantage in terms of protein degradability. It is possible that grazing promotes a high proportion of non-senescent to senescent leaf material within the sward and thus any advantage conferred by the stay-green phenotype would be effectively masked by an abundance of mature foliage. It is suggested that the stay-green trait would be of benefit in areas where agricultural practice permits extensive natural senescence to occur.

  19. Supplementing energy and protein source at different rate of degradability to mixture of corn waste and coffee pod as basal diet on rumen fermentation kinetic of beef cattle

    Directory of Open Access Journals (Sweden)

    Dicky Pamungkas

    2010-03-01

    Full Text Available The use of corn waste and coffee pod as basal diet needs energy and protein supplementation in order to optimize rumen microbial growth. A research was done to study the appropriate supplement which is suitable based on the result of rumen fermentation kinetics. Four ruminally canulated cows, (205-224 kg of live weight were placed in individual cages. The basal diet (BD offered were corn waste and coffee pod mixture (80:20. Source of high degradable energy (HDE was cassava pomace; while the low degradable energy (LDE was arenga piñata waste. The high degradable protein (HDP was mixed-concentrate while the low degradable protein (LDP was leucaena leave meal. The supplementation of energy and protein to basal diet was in ratio of 50:50 based on dry matter. Feed offered were basal diet and the supplement at ratio of 60: 40 (3 % of LW. Observation was carried out for 4 periods (10 days/period. In each period, animal was fed one of the following diets: A = BD, B = BD+ HDE+LDP, C = BD+LDE+LDP, and D = BD+HDE+HDP. Rumen kinetics observed were: pH, and VFA, NH3 and rumen microbial protein concentrations. Rumen fluid was taken at the end of each period gradually along the course of 24 hours fermentation. The results showed that the diets gave significant effect (P < 0.05 on rumen pH. The lowest rumen pH (5.76 was observed on diet D at 2:00, 12 hours after feeding. Meanwhile, the highest pH (7.22 was found in animal fed diet A at 16:00. The total VFA on diet D of periode:1, 3 and 4 showed the highest level: 68.1 mmol/l; 75.37 mmol/l and 85.14 mmol/l respectively. The highest NH3 concentration was found in diet D followed by diet C, B and A. At 12:00 observation or at 4 h after morning feeding the highest NH3 was observed from diet D (41.94 mg/100 ml. It is concluded that diet D resulted in the best rumen fermentation kinetic, therefore it could be used in feed formulation in cattle diets.

  20. Chemical approaches to targeted protein degradation through modulation of the ubiquitin-proteasome pathway.

    Science.gov (United States)

    Collins, Ian; Wang, Hannah; Caldwell, John J; Chopra, Raj

    2017-03-15

    Manipulation of the ubiquitin-proteasome system to achieve targeted degradation of proteins within cells using chemical tools and drugs has the potential to transform pharmacological and therapeutic approaches in cancer and other diseases. An increased understanding of the molecular mechanism of thalidomide and its analogues following their clinical use has unlocked small-molecule modulation of the substrate specificity of the E3 ligase cereblon (CRBN), which in turn has resulted in the advancement of new immunomodulatory drugs (IMiDs) into the clinic. The degradation of multiple context-specific proteins by these pleiotropic small molecules provides a means to uncover new cell biology and to generate future drug molecules against currently undruggable targets. In parallel, the development of larger bifunctional molecules that bring together highly specific protein targets in complexes with CRBN, von Hippel-Lindau, or other E3 ligases to promote ubiquitin-dependent degradation has progressed to generate selective chemical compounds with potent effects in cells and in vivo models, providing valuable tools for biological target validation and with future potential for therapeutic use. In this review, we survey recent breakthroughs achieved in these two complementary methods and the discovery of new modes of direct and indirect engagement of target proteins with the proteasome. We discuss the experimental characterisation that validates the use of molecules that promote protein degradation as chemical tools, the preclinical and clinical examples disclosed to date, and the future prospects for this exciting area of chemical biology. © 2017 The Author(s).

  1. Rumen degradability and ileal digestibility of proteins and amino acids of feedstuffs for cows

    Directory of Open Access Journals (Sweden)

    Iveta Maskaľová

    2014-01-01

    Full Text Available Knowledge of the profile of amino acids of the rumen-undegradable protein can help in the formulation of diets to provide amino acids that complement microbial protein as well as supply amino acids, which are most limiting for milk production. Three non-lactating cows fitted with rumen cannulas were used to determine the effect of in situ rumen degradation on crude protein and amino acid profile of rumen-undegraded protein of feedstuffs. The obtained values of rumen degradability of crude protein with significant difference (P in vitro modified 3-step method was used to determine intestinal digestibility. Intestinal digestibility of undegraded protein varied from 54.5 ± 1.4% in raw soybean to 95.2 ± 1.0% in corn gluten feed. The absorbable amino acid profile of rumen-undegraded protein for each feedstuff was compared with profiles of the original feedstuff and the rumen-exposed undegraded protein. Absorbable lysine (9.3 ± 1.1 g/kg of crude protein was higher in products of soybean and sunflower cake. Corn gluten feed and meal supplied more absorbable methionine (3.6 ± 0.6 g/kg of crude protein. This study showed that the digestibility factor of crude protein and amino acid based on in situ and in vitro methods for thermal treatment of protein feeds can be used in models to optimize the amino acid nutrition of dairy cows and expand knowledge about rumen degradability and ileal digestibility of amino acids in feedstuffs.

  2. 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.

  3. Influência da monensina no consumo e na fermentação ruminal em bovinos recebendo dietas com teores baixo e alto de proteína Effects of different dietary levels of monensin and protein on intake and ruminal fermentation in bovines

    Directory of Open Access Journals (Sweden)

    Marcus Vinicius Morais de Oliveira

    2005-10-01

    Full Text Available Foram utilizados quatro novilhos holandeses fistulados no rúmen e alimentados quatro vezes ao dia (8, 11, 14 e 17 h com dietas contendo teores baixo e alto de proteína (baixo e alto, com e sem monensina, totalizando quatro dietas experimentais (tratamentos, com o intuito de se verificar a influência da monensina sobre o consumo e a fermentação ruminal. As dietas contendo teores de proteína possuíam, respectivamente, 11,4 e 16,5% de proteína bruta na matéria seca e eram constituídas por 65% de feno de capim-braquiária (Brachiaria decumbens e 35% de concentrado, sendo o concentrado da dieta com baixo teor protéico à base de milho e uréia e o da dieta com alto teor protéico à base de milho e farelo de soja. O nível de ionóforo utilizado foi de 28 mg de monensina/kg de MS consumida. As amostras de líquido ruminal foram coletadas diariamente às 13 h. Utilizou-se delineamento em quadrado latino, com quatro tratamentos (dietas e quatro animais. As dietas com alto teor protéico proporcionaram aumento da concentração ruminal do ácido butírico e da amônia. O fornecimento de monensina sódica, independentemente do teor protéico das dietas, promoveu diminuição no consumo de matéria seca, aumento na concentração de ácido propiônico e redução do teor de ácido butírico, da relação acetato:propionato e da atividade específica de produção de amônia. A monensina, quando associada à dieta com baixo teor protéico, também ocasionou diminuição da concentração do ácido acético e elevação do pH e da síntese de proteína microbiana ruminal. Não foram observadas diferenças estatísticas significativas na concentração de amônia ruminal com a inclusão de monensina.Four ruminally fistulated Holstein steers were fed four times a day (8 a.m., 11 a.m., 2 p.m. and 5 p.m. with four diets with low and high protein content, with or without monensin, to verify the effect of monensin on feed intake and ruminal

  4. Short communication: grain-induced subacute ruminal acidosis is associated with the differential expression of insulin-like growth factor-binding proteins in rumen papillae of lactating dairy cattle.

    Science.gov (United States)

    Steele, M A; Alzahal, O; Walpole, M E; McBride, B W

    2012-10-01

    The objective of this study was to characterize the mRNA expression of genes involved in the insulin-like growth factor (IGF) axis in the rumen epithelium during grain-induced ruminal acidosis. Eight lactating dairy cattle were randomly assigned to a control (38% concentrate) or a high-grain (HG; 57% concentrate) diet in a randomized study. Dry matter intake, milk production, ruminal pH, and rumen papillae gene expression were measured before treatment allocation (d 0) and on the fourth day of treatment. On d 4, no differences were observed in total feed intake and milk production; however, the cattle fed the HG diet displayed lower ruminal pH (587 ± 130 min/d below 5.6; mean ± SE) compared with cattle receiving the control diet (169 ± 145 min/d below 5.6). No change in the relative mRNA expression of IGF-1, IGF-1 receptor (IGF-1R), and IGF-binding protein 6 (IGFBP6) was detected between treatments. However, the relative expression value of IGF-binding protein 3 (IGFBP3) decreased (0.73 ± 0.07 fold, mean ± SE), whereas IGF-binding protein 5 (IGFBP5) expression increased (1.53 ± 0.20 fold). These results indicate that the IGF axis may play a role in rumen epithelial adaptation to HG diets. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  5. Iron-Binding Protein Degradation by Cysteine Proteases of Naegleria fowleri.

    Science.gov (United States)

    Martínez-Castillo, Moisés; Ramírez-Rico, Gerardo; Serrano-Luna, Jesús; Shibayama, Mineko

    2015-01-01

    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.

  6. 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.

  7. Investigation of the use of various plant extracts activity in ruminant

    Science.gov (United States)

    Yüca, Songül; Gül, Mehmet; Ćaǧlayan, Alper

    2016-04-01

    The prohibition of the use of antibiotics and as a result of the adverse effect on health of synthetic products, research has focused on natural feed additives. In recent years, the diet of farm animals many feed additives have been used for various purposes or continues. These include as used in ruminant rations as plant extract thyme, anise, pepper, mint, garlic, rosemary, cinnamon, parsley, bay leaf, coconut, like used herbal extracts and their effects on the performance of ruminants was investigated. Antioxidant, antifungal, antiviral, anti-inflamaotry is known to have effects of plant extract. By stimulating the digestive system of ruminants, they increase the activity of digestive enzymes, to prevent environmental pollution caused by manure, regulations rumen fermentation, inhibition of methane formation and protein degradability in the rumen as well as the animal is known to have many benefits. The structure of essential oils and plant extracts in this collection, examining the use of ruminant livestock events and the importance of the use in animal nutrition into practice will be discussed.

  8. DEGRADABILIDADE IN SITU DA MATÉRIA SECA, DA PROTEÍNA BRUTA E DA FRAÇÃO FIBROSA DE CONCENTRADOS E SUBPRODUTOS AGROINDUSTRIAIS IN SITU DEGRADABILITY OF DRY MATTER, CRUDE PROTEIN AND FIBROUS FRACTION OF CONCENTRATE AND AGROINDUSTRIAL BY-PRODUCTS

    Directory of Open Access Journals (Sweden)

    Gleidson Giordano Pinto de Carvalho Carvalho

    2009-09-01

    Full Text Available Objetivou-se avaliar a degradabilidade ruminal da matéria seca (MS, da proteína bruta (PB, da fibra em detergente neutro (FDN e da fibra em detergente ácido (FDA do milho (Zea mays, do farelo de soja (Glicyne max L., da torta de dendê (Elaeis guineensis Jacq. e do farelo de cacau (Theobroma cacao L.. Incubaram-se amostras de cada alimento no rúmen de três novilhos por períodos de 0; 3; 6; 12; 24 e 48 horas. As degradabilidades efetivas da MS, PB, FDN e FDA, para a taxa de passagem de 5%/hora, foram relativamente baixas (abaixo de 60%, exceto para a PB do farelo de soja (acima de 65%. O farelo de soja apresentou os maiores coeficientes de degradação, tanto para MS e PB como também para os constituintes da parede celular, seguido do milho, torta de dendê e farelo de cacau. O farelo de cacau apresentou as menores taxas de degradação ruminal.

    PALAVRAS-CHAVES: Farelo de cacau, incubação ruminal, torta de dendê. The objective of the experiment was to evaluate the dry matter (DM, crude protein (CP, neutral detergent fiber (NDF and acid detergent fiber (ADF ruminal degradability of corn (Zea mays, soybean meal (Glicyne max L., palm kernel cake (Elaeis guineensis Jacq. and cocoa meal (Theobroma cacao L.. Samples of each feed were incubated in rumens of three steers for periods of 0; 3; 6; 12; 24 and 48 hours. The DM, CP, NDF and ADF effective degradabilities, for a passage rate of 5%/hour, were relatively low (lower than 60%, except for soybean meal CP (higher than 65%. Soybean meal showed the greatest degradation coefficients for DM and CP as so as for cellular wall constituents, followed by corn, palm kernel cake and cocoa meal. Cocoa meal showed the lowest ruminal degradation rates.

    KEY WORDS: Cocoa meal, incubation ruminal, palm kernel cake.

  9. Chemical composition of alfalfa silage with waste date and its feeding effect on ruminal fermentation characteristics and microbial protein synthesis in sheep.

    Science.gov (United States)

    Rajabi, R; Tahmasbi, R; Dayani, O; Khezri, A

    2017-06-01

    This study was conducted to evaluate the effect of feeding ensiled alfalfa with waste date on ruminal fermentation characteristics, microbial protein synthesis, protozoa population and blood parameters in sheep. Eight rams were used in a 2 × 2 change over design. Each experimental period consisted of 21 days including 16 days for adaptation and 5 days for sampling. For ensiling, fresh alfalfa (Medicago sativa L.) with different levels of waste date (Phoenix dactylifera L.) were mixed together and ensiled in 100-l containers for 45 days. Chemical composition of silages such as dry matter (DM), crude protein, NH3 -N, organic matter, NDF, ADF and pH were determined. Then, it was used as 30% (DM basis) in diets. The experimental diets were as follows: (i) control (diet containing alfalfa silage without waste date), (ii) diet containing alfalfa silage with 5 g waste date/95 g DM, (iii) diet containing alfalfa silage with 10 g waste date/90 g DM, and (iv) diet containing alfalfa silage with 15 g waste date/85 g DM. The results of this experiment showed that adding waste date to alfalfa during ensiling, improved silage quality, DM and energy level. Total protozoa population and all of holotrich, cellulolytic and entodinia in rumen fluid were increased linearly by increasing the level of waste date. Nitrogen (N) intake, urinary N excretion and nitrogen retention were affected by dietary treatments. Also, allantoin, uric acid, total purine derivatives, microbial protein synthesis, cholesterol level and blood urea nitrogen were significantly different. In conclusion, direct ensilage of alfalfa can be attained by mixing 15 g waste date/85 g DM and positive associative effects such as increased metabolizable energy (ME) and silage quality occurred. Journal of Animal Physiology and Animal Nutrition © 2016 Blackwell Verlag GmbH.

  10. A unique mechanism for protein processing and degradation in Arabidopsis thaliana

    Science.gov (United States)

    Rojo, Enrique; Zouhar, Jan; Carter, Clay; Kovaleva, Valentina; Raikhel, Natasha V.

    2003-01-01

    Precursor protease vesicles are plant-specific compartments containing precursors of enzymes that are thought to participate in the degradation of cellular components in organs undergoing senescence. We report in vivo evidence that the precursor protease vesicle-localized vacuolar processing enzyme-γ (VPEγ) is critical for maturation of the plant vacuolar protease AtCPY. We also provide biochemical and functional evidence that VPEγ is involved in degradation of the vacuolar invertase AtFruct4 in aging tissues. Moreover, a proteomics-based approach identified various proteins found in the vacuoles of aging vpeγ mutants but not in WT plants, suggesting a unique role of VPEγ in protein processing and degradation in Arabidopsis. PMID:12773619

  11. Ubiquitin ligase gp78 targets unglycosylated prion protein PrP for ubiquitylation and degradation.

    Directory of Open Access Journals (Sweden)

    Jia Shao

    Full Text Available 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 ligase gp78, known for its role in protein quality control, is critical for unglycosylated PrP ubiquitylation and degradation. Furthermore, C-terminal sequences of PrP protein are crucial for its ubiquitylation and degradation. Our study reveals the first ubiquitin ligase specifically involved in prion protein PrP degradation and PrP sequences crucial for its turnover. Our data may lead to a new avenue to control PrP level and pathogenesis.

  12. 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...

  13. Degradation and de novo synthesis of D1 protein and psbA ...

    Indian Academy of Sciences (India)

    synthesis than the visible radiation (Jones and Kok 1966). As compared to vast literature available on visible ..... 1997). The degradation of other PS II core proteins including CP 47 and change in heterogeneity can not be ruled out. The apparent photosynthesis that observed in. UV-B treated C. reinhardtii is the net result of ...

  14. Effect of cleaning agents and additives on Protein A ligand degradation and chromatography performance.

    Science.gov (United States)

    Yang, Lihua; Harding, Jason D; Ivanov, Alexander V; Ramasubramanyan, Natarajan; Dong, Diane D

    2015-03-13

    Protein A chromatography, employing the recombinant Protein A ligand, is widely used as a capture step for antibody and Fc-fusion proteins manufacture. Protein A ligands in these matrices are susceptible to degradation/loss when exposed to cleaning agents such as sodium hydroxide, resulting in loss of capacity on reuse. In this study, MabSelect Protein A ligand and MabSelect SuRe Protein A ligand were chosen to evaluate the impact of alkaline cleaning solutions on the ligands and the packed columns. The Protein A ligands alone and the Protein A columns were incubated or cycled in different concentrations of sodium hydroxide solutions with and without additives, respectively. Ligand integrity (degradation) and ligand function (binding affinity) were studied using SDS-PAGE and customized Biacore technology, surface plasma resonance (SPR) and were successfully correlated with column performance measurement in terms of static binding capacity (SBC), dynamic binding capacity (DBC) and recovery as a function of exposure to cleaning agents with and without additives. The findings and the methodology presented in this study are not only able to determine appropriate cleaning conditions for Protein A chromatography, but also provided tools to enable systematic and rapid study of the cleaning solutions and conditions. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. 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

  16. peste des petits ruminants

    African Journals Online (AJOL)

    HP USER

    of antibodies to peste des petits ruminants. (PPR) virus. Veterinary. Research. Communications, 30:325-330. Singh RK, Balauirjgal V, Bhamuprakash V, Sanen A,. Saravanan P, & Yadev MP (2009). Possible control and eradication of peste des petits ruminants from India: technical aspect. Veterinaria Italiana, 45 (3), ...

  17. A comparison of different legume seeds as protein supplement to optimise the use of low quality forages by ruminants

    DEFF Research Database (Denmark)

    Yánez-Ruiz, David R; Martin-Garcia, Antonio I; Weisbjerg, Martin Riis

    2009-01-01

    The potential of different legume seeds species, including recently new developed varieties (Vicia faba: a commercial variety and varieties Alameda, Palacio and Baraka; Lupinus angustifolius; Pisum sativum and Cicer arietinum: varieties Fardon and Zegr ) as protein supplements to low quality...

  18. Consumo e fermentação ruminal de proteínas em função de suplementação alimentar energética e protéica em novilhos Feed intake and ruminal fermentation of proteins as a function of energetic and proteic food supplementation of steers

    Directory of Open Access Journals (Sweden)

    Natália Guarino Souza Barbosa

    2001-10-01

    Full Text Available Objetivou-se analisar o efeito de suplementação energética sobre o consumo e a fermentação ruminal de proteínas em dois níveis de proteína dietética (6 e 20%. Quatro novilhos mestiços fistulados no rúmen foram utilizados em um quadrado latino e os tratamentos arranjados em um fatorial 2 × 2, em que as dietas testadas constituíram-se de: A. só feno; B. feno + 2,8 kg de farelo de soja; C. feno + 2,8 kg de fubá de milho; e D. feno + 2,8 kg de farelo de soja + 2,8 kg de fubá de milho. Foram realizados quatro períodos experimentais constituídos de 18 dias, sendo sete dias para adaptação dos animais, sete dias para determinação do consumo de matéria seca e quatro dias para coleta de amostras. As coletas foram feitas através de fístula ruminal nos tempos zero, dois, quatro e seis horas após a alimentação, sendo utilizadas para determinações das concentrações de amônia, pH e AGV do líquido ruminal. Não houve interações entre o fubá de milho e o farelo de soja, assim como não ocorreu efeito do tempo de coleta sobre os parâmetros estudados. O farelo de soja reduziu a relação acetato:propionato (A:P em 13%, aumentou o consumo de matéria seca total (CMSt em 41,2%, sem alterar o consumo de forragem e o pH, e aumentou as concentrações de amônia (790%, propionato (48%, AGV total (39%, isobutirato (165%, isovalerato (208% e valerato (201%. O fubá de milho não afetou a fermentação de proteína, embora tenha causado aumento no CMSt (35% e decréscimo no pH ruminal (6% e na relação A:P (18%.The objective was to evaluate the effect of energetic supplementation on feed intake and ruminal fermentation of proteins in two protein levels in the diet (6 and 20%. There were used four rumen fistulated crossbred steers in a Latim square and the treatments were arranjed in a 2 × 2 factorial, in which the combinations were: A. only hay; B. hay + 2.8 kg of soybean meal; C. hay + 2.8 kg of corn meal; and D. hay + 2.8 kg of

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

    Directory of Open Access Journals (Sweden)

    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.

  20. Estimativa da degradabilidade ruminal de alimentos utilizando a técnica de produção de gás em bovinos, ovinos e caprinos Estimate of the ruminal degradability of some feeds using gas production technique in cattle, sheep, and goats

    Directory of Open Access Journals (Sweden)

    Roseli Aparecida dos Santos

    2003-06-01

    Full Text Available Conduziu-se este trabalho com o objetivo de validar a técnica de produção de gás na avaliação de alimentos concentrados e volumosos e as possíveis diferenças entre bovinos, ovinos e caprinos quanto à degradação desses alimentos. O experimento foi conduzido no Laboratório de Pesquisa Animal do Departamento de Zootecnia da UFLA. O líquido ruminal (inóculo foi originado de três vacas Holandesas, três ovelhas e três cabras sem raça definida, fistuladas no rúmen. Foram avaliadas as frações solúvel em detergente neutro (SDN e fibra em detergente neutro(FDN de fubá de milho, farelo de soja, farelo de algodão, caroço de algodão, farelo de trigo, polpa cítrica, feno de alfafa, feno de coast cross, silagem de milho e silagem de capim. As leituras do volume cumulativo de gás foram feitas 1, 2, 3, 4, 5, 6, 12, 18, 24, 30, 36, 48, 60 e 72 horas após a incubação. Não houve diferença (P>0,05 entre as espécies, para nenhuma das variáveis estudadas. A fração SDN apresentou maior produção de gás e taxa de degradação em relação a FDN. A técnica de produção de gás permitiu estimar as taxas de digestão das frações insolúveis e, principalmente, das frações solúveis dos carboidratos totais.The objective of this work was to validate the technique of gas production in the evaluation of concentrated feeds and roughages, and the possible differences between cattle, sheep, and goats, regarding the degradation of these feeds. The experiment was carried out in the Laboratory of Animal Research at the Animal Science Department - Federal University of Lavras, Brazil. The ruminal liquor came from three Holstein cows, three sheep, and three goats, without defined race. All were fistulated in the rumen. The neutral detergent soluble fraction (NDS and neutral detergent fiber (NDF, of ground corn, soybean meal, cotton meal, cotton seed, wheat meal, citrus pulp, alfalfa hay, coast cross hay, corn silage and grass silage were

  1. 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.

  2. Effects of ruminally undegradable protein levels on nitrogen and phosphorus balance and their excretion in Saanen goats fed oil palm fronds

    Directory of Open Access Journals (Sweden)

    Basery, M.

    2004-01-01

    Full Text Available Twelve Saanen goats 3-4 months old with initial live weight (mean±SD of 18.8±2.2 kg were used in a 4×4 Latin square design. Diets were isonitrogenous (15% CP and isocaloric (3.6 Mcal/kg DM and were fed ad libitum. Total mixed diets were composed of steamed oil palm fronds (OPF and concentrate [urea, soybean meal (SBM, cassava waste, molasses, sulphur and commercial mineral and vitamin mix]. The treatments were four levels of ruminally undegradable protein (RUP supplementation viz, 0, 2, 4 or 6% (0%RUP, 2%RUP, 4%RUP or 6%RUP, respectively. Nitrogen (N and phosphorus (P intakes tended to be increased linearly (p=0.08 and p=0.09 as a consequence of additional of RUP supplementation. Similarly, %N absorption and retention, dry matter intake (DMI and crude protein digestibility increased linearly (p>0.05 as a consequence of the additional of RUP supplementation. There were no effects of dietary treat- ment on P balance. However, P intake, absorption and retention tended to be increased as a consequence of additional of RUP supplementation. The linear equation between RUP (% and P intake (g/d of goats was Pintake = 2.05+0.056RUP; (p=0.07. Nitrogen and P retention also tended to increase with increasing levels of RUP supplementation. Manipulation of diet by supplementation of RUP in the rations can be used to reduce the amount of N excreted, especially N in urine, which in turn reduces ammonia emissions.

  3. Degradation of the disease-associated prion protein by a serine protease from lichens

    Science.gov (United States)

    Johnson, Christopher J.; Bennett, James P.; Biro, S.M.; Duque-Velasquez, J. C.; Rodriguez, Cynthia M.; Bessen, R.A.; Rocke, Tonie E.

    2011-01-01

    The disease-associated prion protein (PrPTSE), 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 PrPTSE 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 PrPTSE. 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 PrPTSE-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 PrPTSE 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.

  4. The C-terminal sequence of RhoB directs protein degradation through an endo-lysosomal pathway.

    Directory of Open Access Journals (Sweden)

    Dolores Pérez-Sala

    Full Text Available BACKGROUND: Protein degradation is essential for cell homeostasis. Targeting of proteins for degradation is often achieved by specific protein sequences or posttranslational modifications such as ubiquitination. METHODOLOGY/PRINCIPAL FINDINGS: By using biochemical and genetic tools we have monitored the localization and degradation of endogenous and chimeric proteins in live primary cells by confocal microscopy and ultra-structural analysis. Here we identify an eight amino acid sequence from the C-terminus of the short-lived GTPase RhoB that directs the rapid degradation of both RhoB and chimeric proteins bearing this sequence through a lysosomal pathway. Elucidation of the RhoB degradation pathway unveils a mechanism dependent on protein isoprenylation and palmitoylation that involves sorting of the protein into multivesicular bodies, mediated by the ESCRT machinery. Moreover, RhoB sorting is regulated by late endosome specific lipid dynamics and is altered in human genetic lipid traffic disease. CONCLUSIONS/SIGNIFICANCE: Our findings characterize a short-lived cytosolic protein that is degraded through a lysosomal pathway. In addition, we define a novel motif for protein sorting and rapid degradation, which allows controlling protein levels by means of clinically used drugs.

  5. 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…

  6. Identification of disulfide bonds in protein proteolytic degradation products using de novo-protein unique sequence tags approach.

    Science.gov (United States)

    Shen, Yufeng; Tolić, Nikola; Purvine, Samuel O; Smith, Richard D

    2010-08-06

    Disulfide bonds are a form of post-translational modification that often determines protein structure(s) and function(s). In this work, we report a mass spectrometry method for identification of disulfides in degradation products of proteins, specifically endogenous peptides in the human blood plasma peptidome. LC-Fourier transform tandem mass spectrometry (FT MS/MS) was used for acquiring mass spectra that were de novo sequenced and then searched against the IPI human protein database. Through the use of unique sequence tags (UStags), we unambiguously correlated the spectra to specific database proteins. Examination of the UStags' prefix and/or suffix sequences that contain cysteine(s) in conjunction with sequences of the UStags-specified database proteins is shown to enable the unambigious determination of disulfide bonds. Using this method, we identified the intermolecular and intramolecular disulfides in human blood plasma peptidome peptides that have molecular weights of up to approximately 10 kDa.

  7. Involvement of Two Latex-Clearing Proteins during Rubber Degradation and Insights into the Subsequent Degradation Pathway Revealed by the Genome Sequence of Gordonia polyisoprenivorans Strain VH2

    Science.gov (United States)

    Hiessl, Sebastian; Schuldes, Jörg; Thürmer, Andrea; Halbsguth, Tobias; Bröker, Daniel; Angelov, Angel; Liebl, Wolfgang; Daniel, Rolf

    2012-01-01

    The increasing production of synthetic and natural poly(cis-1,4-isoprene) rubber leads to huge challenges in waste management. Only a few bacteria are known to degrade rubber, and little is known about the mechanism of microbial rubber degradation. The genome of Gordonia polyisoprenivorans strain VH2, which is one of the most effective rubber-degrading bacteria, was sequenced and annotated to elucidate the degradation pathway and other features of this actinomycete. The genome consists of a circular chromosome of 5,669,805 bp and a circular plasmid of 174,494 bp with average GC contents of 67.0% and 65.7%, respectively. It contains 5,110 putative protein-coding sequences, including many candidate genes responsible for rubber degradation and other biotechnically relevant pathways. Furthermore, we detected two homologues of a latex-clearing protein, which is supposed to be a key enzyme in rubber degradation. The deletion of these two genes for the first time revealed clear evidence that latex-clearing protein is essential for the microbial utilization of rubber. Based on the genome sequence, we predict a pathway for the microbial degradation of rubber which is supported by previous and current data on transposon mutagenesis, deletion mutants, applied comparative genomics, and literature search. PMID:22327575

  8. Bromatological characteristics and ruminal digestibility of grain corn hybrids with different vitreousness in silage maturity

    Directory of Open Access Journals (Sweden)

    Evandrei Santos Rossi

    2016-06-01

    Full Text Available The aim of this research was to evaluate vitreousness in commercial corn hybrids with flint and dent grains and to study the effect of this characteristic on agronomic behavior, nutritional value, and ruminal degradability of grains harvested in silage maturity. Twelve commercial corn hybrids were evaluated. They were divided in two groups (six with flint grains and six with dent grains. The experiments were conducted in two municipalities represented by Guarapuava and Laranjeiras do Sul, both in Paraná State. The harvest for the grain quality analysis was performed at ¾ of the milk line stage in the grain. The following characteristics were evaluated: grain yield, grain yield in silage maturity, vitreousness, ruminal digestibility of grain, neutral detergent fiber, acid detergent fiber and crude protein. The grains in the group of dent hybrids exhibited better degradability associated with lower vitreousness than the group of flint hybrids. The group of dent hybrids exhibited better ruminal digestibility of the grains associated with lower vitreousness versus the group of flint hybrids. There is a negative correlation between vitreousness and the digestibility of the grain; thus, vitreousness can be a criterion for selecting genotypes for forage production.

  9. 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.

  10. Mitochondrial protein oxidation and degradation in response to oxidative stress and aging.

    Science.gov (United States)

    Bulteau, Anne-Laure; Szweda, Luke I; Friguet, Bertrand

    2006-07-01

    Mitochondria are a major source of intracellular reactive oxygen species (ROS), the production of which increases with age. These organelles are also targets of oxidative damage. The deleterious effects of ROS may be responsible for impairment of mitochondrial function observed during various pathophysiological states associated with oxidative stress and aging. An important factor for protein maintenance in the presence of oxidative stress is enzymatic reversal of oxidative modifications and/or protein degradation. Failure of these protein maintenance systems is likely a critical component of the aging process. Mitochondrial matrix proteins are sensitive to oxidative inactivation and oxidized proteins are known to accumulate during aging. The ATP-stimulated mitochondrial Lon protease is a highly conserved protease found in prokaryotes and the mitochondrial compartment of eukaryotes and is believed to play an important role in the degradation of oxidized mitochondrial matrix proteins. Age-dependent declines in the activity and regulation of this proteolytic system may underlie accumulation of oxidatively modified and dysfunctional protein and loss in mitochondrial viability.

  11. 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.

  12. 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.

  13. Individual and Collective Contributions of Chaperoning and Degradation to Protein Homeostasis in E. coli

    Directory of Open Access Journals (Sweden)

    Younhee Cho

    2015-04-01

    Full Text Available The folding fate of a protein in vivo is determined by the interplay between a protein’s folding energy landscape and the actions of the proteostasis network, including molecular chaperones and degradation enzymes. The mechanisms of individual components of the E. coli proteostasis network have been studied extensively, but much less is known about how they function as a system. We used an integrated experimental and computational approach to quantitatively analyze the folding outcomes (native folding versus aggregation versus degradation of three test proteins biosynthesized in E. coli under a variety of conditions. Overexpression of the entire proteostasis network benefited all three test proteins, but the effect of upregulating individual chaperones or the major degradation enzyme, Lon, varied for proteins with different biophysical properties. In sum, the impact of the E. coli proteostasis network is a consequence of concerted action by the Hsp70 system (DnaK/DnaJ/GrpE, the Hsp60 system (GroEL/GroES, and Lon.

  14. Protection of a protein against irradiation-induced degradation by additives in the solid state

    Science.gov (United States)

    Shalaev, E.; Reddy, R.; Kimball, R. N.; Weinschenk, M. F.; Guinn, M.; Margulis, L.

    2003-03-01

    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.

  15. Preliminary results on the effects of grape (Vitis vinifera) seed condensed tannins on in vitro intestinal digestibility of the lupin (Lupinus angustifolius) seed protein fraction in small ruminants.

    Science.gov (United States)

    Bruno-Soares, A M; Soares-Pereira, A L; Matos, T J S; Ricardo-da-Silva, J M

    2011-08-01

    Condensed tannins (CT) from grape seeds (Vitis vinifera L.) were added to complex the protein fraction of Lupinus angustifolius seeds. Three CT/protein ratios were used: 96 mg/g (T(1)), 180 mg/g (T(2)) and 0 mg/g (T(0)). The CP losses in the rumen were assessed by the nylon-bag technique and CP intestinal digestibility (CPID) was estimated using an in vitro assay applying a three-step procedure: samples were subject to rumen degradation (in situ, 16 h) and the remaining residues were subject to the digestive enzymes of the abomasum and pancreas in vitro. A positive effect (p 0.05) by the presence of CT. These findings suggest that the use of grape seed CT might have the potential to improve the efficiency of utilisation of the protein fraction from lupin seeds. © 2010 Blackwell Verlag GmbH.

  16. Different Stability and Proteasome-Mediated Degradation Rate of SMN Protein Isoforms.

    Directory of Open Access Journals (Sweden)

    Denise Locatelli

    Full Text Available The key pathogenic steps leading to spinal muscular atrophy (SMA, a genetic disease characterized by selective motor neuron degeneration, are not fully clarified. The full-length SMN protein (FL-SMN, the main protein product of the disease gene SMN1, plays an established role in the cytoplasm in snRNP biogenesis ultimately leading to mRNA splicing within the nucleus. It is also involved in the mRNA axonal transport. However, to what extent the impairment of these two SMN functions contributes to SMA pathogenesis remains unknown. A shorter SMN isoform, axonal-SMN or a-SMN, with more specific axonal localization, has been discovered, but whether it might act in concert with FL-SMN in SMA pathogenesis is not known. As a first step in defining common or divergent intracellular roles of FL-SMN vs a-SMN proteins, we here characterized the turn-over of both proteins and investigated which pathway contributed to a-SMN degradation. We performed real time western blot and confocal immunofluorescence analysis in easily controllable in vitro settings. We analyzed co-transfected NSC34 and HeLa cells and cell clones stably expressing both a-SMN and FL-SMN proteins after specific blocking of transcript or protein synthesis and inhibition of known intracellular degradation pathways. Our data indicated that whereas the stability of both FL-SMN and a-SMN transcripts was comparable, the a-SMN protein was characterized by a much shorter half-life than FL-SMN. In addition, as already demonstrated for FL-SMN, the Ub/proteasome pathway played a major role in the a-SMN protein degradation. We hypothesize that the faster degradation rate of a-SMN vs FL-SMN is related to the protection provided by the protein complex in which FL-SMN is assembled. The diverse a-SMN vs FL-SMN C-terminus may dictate different protein interactions and complex formation explaining the different localization and role in the neuronal compartment, and the lower expression and stability of a-SMN.

  17. Global analysis of phosphorylation and ubiquitylation cross-talk in protein degradation.

    Science.gov (United States)

    Swaney, Danielle L; Beltrao, Pedro; Starita, Lea; Guo, Ailan; Rush, John; Fields, Stanley; Krogan, Nevan J; Villén, Judit

    2013-07-01

    Cross-talk between different types of post-translational modifications on the same protein molecule adds specificity and combinatorial logic to signal processing, but it has not been characterized on a large-scale basis. We developed two methods to identify protein isoforms that are both phosphorylated and ubiquitylated in the yeast Saccharomyces cerevisiae, identifying 466 proteins with 2,100 phosphorylation sites co-occurring with 2,189 ubiquitylation sites. We applied these methods quantitatively to identify phosphorylation sites that regulate protein degradation via the ubiquitin-proteasome system. Our results demonstrate that distinct phosphorylation sites are often used in conjunction with ubiquitylation and that these sites are more highly conserved than the entire set of phosphorylation sites. Finally, we investigated how the phosphorylation machinery can be regulated by ubiquitylation. We found evidence for novel regulatory mechanisms of kinases and 14-3-3 scaffold proteins via proteasome-independent ubiquitylation.

  18. Primary transmission of chronic wasting disease versus scrapie prions from small ruminants to transgenic mice expressing ovine or cervid prion protein.

    Science.gov (United States)

    Madsen-Bouterse, Sally A; Schneider, David A; Zhuang, Dongyue; Dassanayake, Rohana P; Balachandran, Aru; Mitchell, Gordon B; O'Rourke, Katherine I

    2016-09-01

    Development of mice expressing either ovine (Tg338) or cervid (TgElk) prion protein (PrP) have aided in characterization of scrapie and chronic wasting disease (CWD), respectively. Experimental inoculation of sheep with CWD prions has demonstrated the potential for interspecies transmission but, infection with CWD versus classical scrapie prions may be difficult to differentiate using validated diagnostic platforms. In this study, mouse bioassay in Tg338 and TgElk was utilized to evaluate transmission of CWD versus scrapie prions from small ruminants. Mice (≥5 per homogenate) were inoculated with brain homogenates from clinically affected sheep or goats with naturally acquired classical scrapie, white-tailed deer with naturally acquired CWD (WTD-CWD) or sheep with experimentally acquired CWD derived from elk (sheep-passaged-CWD). Survival time (time to clinical disease) and attack rates (brain accumulation of protease resistant PrP, PrPres) were determined. Inoculation with classical scrapie prions resulted in clinical disease and 100 % attack rates in Tg338, but no clinical disease at endpoint (>300 days post-inoculation, p.i.) and low attack rates (6.8 %) in TgElk. Inoculation with WTD-CWD prions yielded no clinical disease or brain PrPres accumulation in Tg338 at endpoint (>500 days p.i.), but rapid onset of clinical disease (~121 days p.i.) and 100 % attack rate in TgElk. Sheep-passaged-CWD resulted in transmission to both mouse lines with 100 % attack rates at endpoint in Tg338 and an attack rate of ~73 % in TgElk with some culled due to clinical disease. These primary transmission observations demonstrate the potential of bioassay in Tg338 and TgElk to help differentiate possible infection with CWD versus classical scrapie prions in sheep and goats.

  19. 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.

  20. Gold nanoparticles enhance the X-ray-induced degradation of human centrin 2 protein

    Science.gov (United States)

    Brun, Emilie; Duchambon, Patricia; Blouquit, Yves; Keller, Gérard; Sanche, Léon; Sicard-Roselli, Cécile

    2009-03-01

    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 2×10 -4 and with the higher X-ray energy of 49 keV.

  1. Pepsin degradation of Cry1A(b) protein purified from genetically modified maize (Zea mays).

    Science.gov (United States)

    de Luis, Ruth; Lavilla, María; Sánchez, Lourdes; Calvo, Miguel; Pérez, María D

    2010-02-24

    The aim of this work was to study the in vitro digestion of Cry1A(b) protein by pepsin. To perform this work, a protein fraction purified from transgenic maize by immunoadsorption was employed. The undigested fraction showed several bands of molecular weight ranging between 14 and 70 kDa when assayed by SDS-PAGE. These bands were identified as corresponding to Cry1A(b) protein by immunochemical techniques and mass spectrometry. The rate of degradation of the purified fraction by pepsin estimated by ELISA was found to be about 75% within 30 min, and the protein concentration remained constant up to 4 h. In all treated samples, the full-length protein and fragments present in Cry1A(b) fraction were absent and peptides of less than 8.5 kDa were mainly found by SDS-PAGE and mass spectrometry. These peptides did not react with antiserum against Cry1A(b) protein by Western blotting. These results suggest that Cry1A(b) fraction purified from transgenic maize is rapidly and extensively degraded by pepsin, giving peptides of low molecular mass.

  2. Effects of ruminal doses of sucrose, lactose, and corn starch on ruminal fermentation and expression of genes in ruminal epithelial cells.

    Science.gov (United States)

    Oba, M; Mewis, J L; Zhining, Z

    2015-01-01

    The objective was to evaluate effects of a ruminal dose of sucrose, lactose, and corn starch on ruminal fermentation and expression of genes in ruminal epithelial cells. Six ruminally cannulated nonlactating nonpregnant Holstein cows (body weight=725±69.6kg) were assigned to treatments in a 3×3 Latin square design with 7-d periods; 1d for data and sample collection followed by a 6-d washout period. Cows were fed a diet containing whole-crop barley silage and dry ground corn, and dietary neutral detergent fiber and crude protein contents were 41.8 and 13.2% [dry matter (DM) basis], respectively. Treatment was a pulse-dose of sucrose, lactose, and corn starch (3.0, 3.0, and 2.85kg of DM, respectively; providing similar amounts of hexose across the treatments) through the ruminal cannulas. All treatments were given with alfalfa silage (1.75kg DM) to prevent acute rumen acidosis. Rumen pH was continuously monitored, and rumen fluid was sampled at 0, 30, 60, 90, 120, 150, and 180min after the dose. In addition, ruminal papillae were sampled from the ventral sac at 180min after the dose. Ruminal dosing with sucrose and lactose, compared with corn starch, increased ruminal total volatile fatty acid concentration and molar proportion of butyrate from 60 to 180min after the dose, and expression of genes for sodium hydrogen exchanger isoforms 1 and 2, and ATPase isoform 1 in ruminal epithelial cells. Ruminal dosing with sucrose, compared with lactose and corn starch, decreased rumen pH from 120 to 180min after the dose and molar proportion of acetate in ruminal fluid from 60 to 150min after the dose, and increased molar proportion of propionate in ruminal fluid from 60 to 150min, and expression of genes involved in butyrate metabolism (3-hydroxy-3-methylglutaryl-coenzyme A synthase isoform 1) and anion exchange across ruminal apical cell membrane (putative anion transporter isoform 1). These results suggest that replacing dietary starch with sugars may affect ruminal

  3. HaloPROTACS: Use of Small Molecule PROTACs to Induce Degradation of HaloTag Fusion Proteins.

    Science.gov (United States)

    Buckley, Dennis L; Raina, Kanak; Darricarrere, Nicole; Hines, John; Gustafson, Jeffrey L; Smith, Ian E; Miah, Afjal H; Harling, John D; Crews, Craig M

    2015-08-21

    Small molecule-induced protein degradation is an attractive strategy for the development of chemical probes. One method for inducing targeted protein degradation involves the use of PROTACs, heterobifunctional molecules that can recruit specific E3 ligases to a desired protein of interest. PROTACs have been successfully used to degrade numerous proteins in cells, but the peptidic E3 ligase ligands used in previous PROTACs have hindered their development into more mature chemical probes or therapeutics. We report the design of a novel class of PROTACs that incorporate small molecule VHL ligands to successfully degrade HaloTag7 fusion proteins. These HaloPROTACs will inspire the development of future PROTACs with more drug-like properties. Additionally, these HaloPROTACs are useful chemical genetic tools, due to their ability to chemically knock down widely used HaloTag7 fusion proteins in a general fashion.

  4. 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.

  5. Effect of Estradiol-17beta on protein synthesis and degradation rates in fused bovine satellite cell cultures.

    Science.gov (United States)

    Kamanga-Sollo, E; White, M E; Hathaway, M R; Weber, W J; Dayton, W R

    2010-07-01

    Although androgenic and estrogenic steroids are widely used to enhance muscle growth and increase feed efficiency in feedlot cattle, their mechanism of action is not well understood. Further, in vivo studies indicate that estradiol (E2) affects muscle protein synthesis and/or degradation, but in vitro results are inconsistent. We have examined the effects of E2 treatment on protein synthesis and degradation rates in fused bovine satellite cell (BSC) cultures. Additionally, to learn more about the mechanisms involved in E2-enhanced muscle growth, we have examined the effects of compounds that interfere with binding of E2 or insulin-like growth factor (IGF)-1 to their respective receptors on E2-induced alterations in protein synthesis and degradation rates in BSC cultures. Treatment of fused BSC cultures with E2 results in a concentration-dependent increase (P < 0.05) in protein synthesis rate and a decrease (P < 0.05) in protein degradation rate. The pure estrogen antagonist ICI 182 780 suppresses (P < 0.05) E2-induced alterations in protein synthesis and degradation in fused BSC cultures. The G-protein coupled receptor (GPR)-30 agonist G1 does not affect either synthesis or degradation rate, which establishes that GPR30 does not play a role in E2-induced alterations in protein synthesis or degradation. JB1, a competitive inhibitor of IGF-1 binding to the Type 1 insulin-like growth factor receptor (IGFR-1), suppresses (P < 0.05) E2-induced alterations in protein synthesis and degradation. In summary, our data show that E2 treatment directly alters both protein synthesis and degradation rates in fused BSC cultures via mechanisms involving both the classical estrogen receptor (ER) and IGFR-1.

  6. Evaluation of feed value of a by-product of pickled radish for ruminants: analyses of nutrient composition, storage stability, and in vitro ruminal fermentation

    Directory of Open Access Journals (Sweden)

    Seoyoung Jeon

    2016-09-01

    Full Text Available Abstract Background By-products of pickled radish (BPR are considered food waste. Approximately 300 g/kg of the total mass of raw materials becomes BPR. Production of pickled radish has grown continuously and is presently about 40,000 metric tons annually in Korea. The objective of the present study was thus to explore the possibility of using BPR as a ruminant feed ingredient. Results BPR contained a large amount of moisture (more than 800 g/kg and ash, and comprised mostly sodium (103 g/kg DM and chloride (142 g/kg DM. On a dry matter basis, the crude protein (CP and ether extract (EE levels in BPR were 75 g/kg and 7 g/kg, respectively. The total digestible nutrient (TDN level was 527 g/kg and the major portion of digestible nutrients was carbohydrate; 88 % organic matter (OM was carbohydrate and 65 % of total carbohydrate was soluble or degradable fiber. The coefficient of variation (CV of nutrient contents among production batches ranged from 4.65 to 33.83 %. The smallest CV was observed in OM, and the largest, in EE. The variation in CP content was relatively small (10.11 %. The storage stability test revealed that storage of BPR at 20 °C (room temperature might not cause spoilage for 4 d, and possibly longer. If BPR is refrigerated, spoilage can be deferred for 21 d and longer. The in vitro ruminal fermentation study showed that substitution of annual ryegrass straw with BPR improved ruminal fermentation, as evidenced by an increase in VFA concentration, DM degradability, and total gas production. Conclusion The major portion of nutrients in BPR is soluble or degradable fiber that can be easily fermented in the rumen without adverse effects, to provide energy to ruminant animals. Although its high sodium chloride content needs to be considered when formulating a ration, BPR can be successfully used as a feed ingredient in a ruminant diet, particularly if it is one component of a total mixed ration.

  7. An RNA element in human interleukin 6 confers escape from degradation by the gammaherpesvirus SOX protein.

    Science.gov (United States)

    Hutin, Stephanie; Lee, Yeon; Glaunsinger, Britt A

    2013-04-01

    Several viruses express factors to silence host gene expression via widespread mRNA degradation. This phenotype is the result of the coordinated activity of the viral endonuclease SOX and the cellular RNA degradation enzyme Xrn1 during lytic Kaposi's sarcoma-associated herpesvirus (KSHV) infection. While most cellular transcripts are highly downregulated, a subset of host mRNA escapes turnover via unknown mechanisms. One of the most prominent escapees is the interleukin 6 (IL-6) mRNA, which accumulates robustly during KSHV lytic infection and is not subjected to SOX-induced degradation. Here we reveal that the IL-6 mRNA contains a dominant, cis-acting ∼100-nucleotide element within its 3' untranslated region (UTR) that renders it directly refractory to cleavage by SOX. This element specifically interacts with a cellular protein complex both in SOX-transfected cells and in KSHV-infected B cells. Using a directed RNA pulldown approach, we identified two components of this complex to be the AU-rich element (ARE) binding proteins AUF1 and HuR. Depletion of these proteins significantly reduced the protective capacity of the IL-6 RNA element in SOX-expressing cells. These findings suggest that SOX activity may be directly counteracted by select RNA regulatory complexes and reveal a novel mechanism contributing to the robust expression of IL-6 during KSHV replication.

  8. High-Risk Human Papillomavirus E7 Proteins Target PTPN14 for Degradation

    Directory of Open Access Journals (Sweden)

    Elizabeth A. White

    2016-09-01

    Full Text Available The major transformation activity of the high-risk human papillomaviruses (HPV is associated with the E7 oncoprotein. The interaction of HPV E7 with retinoblastoma family proteins is important for several E7 activities; however, this interaction does not fully account for the high-risk E7-specific cellular immortalization and transformation activities. We have determined that the cellular non-receptor protein tyrosine phosphatase PTPN14 interacts with HPV E7 from many genus alpha and beta HPV types. We find that high-risk genus alpha HPV E7, but not low-risk genus alpha or beta HPV E7, is necessary and sufficient to reduce the steady-state level of PTPN14 in cells. High-risk E7 proteins target PTPN14 for proteasome-mediated degradation, which requires the ubiquitin ligase UBR4, and PTPN14 is degraded by the proteasome in HPV-positive cervical cancer cell lines. Residues in the C terminus of E7 interact with the C-terminal phosphatase domain of PTPN14, and interference with the E7-PTPN14 interaction restores PTPN14 levels in cells. Finally, PTPN14 degradation correlates with the retinoblastoma-independent transforming activity of high-risk HPV E7.

  9. Assessing essentiality of transketolase in Mycobacterium tuberculosis using an inducible protein degradation system.

    Science.gov (United States)

    Kolly, Gaëlle S; Sala, Claudia; Vocat, Anthony; Cole, Stewart T

    2014-09-01

    Improved genetic tools are required to identify new drug targets in Mycobacterium tuberculosis. To this aim, genetic approaches, targeting either transcription and/or protein degradation, have been developed to appraise gene essentiality and to test the impact of gene silencing on bacterial survival. Here, we successfully combined the Tet-Pip OFF system, which downregulates transcription through the TetR and Pip repressors, with SspB-mediated protein degradation to study depletion of the transketolase encoded by the tkt (rv1449c) gene. We show that depletion of Tkt using the RNA silencing and protein degradation (RSPD) system arrested growth of M. tuberculosis in vitro faster than the Tet-Pip OFF system alone. In addition, we extended the new combined approach to an ex vivo model of M. tuberculosis infection in THP-1 cells. Tkt-depleted bacteria displayed reduced virulence as compared to wild type bacilli, thus confirming the essentiality of the enzyme for intracellular growth. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  10. The Clp Proteases of Bacillus subtilis Are Directly Involved in Degradation of Misfolded Proteins

    OpenAIRE

    Krüger, Elke; Witt, Elke; Ohlmeier, Steffen; Hanschke, Renate; Hecker, Michael

    2000-01-01

    The presence of the heat stress response-related ATPases ClpC and ClpX or the peptidase ClpP in the cell is crucial for tolerance of many forms of stress in Bacillus subtilis. Assays for detection of defects in protein degradation suggest that ClpC, ClpP, and ClpX participate directly in overall proteolysis of misfolded proteins. Turnover rates for abnormal puromycyl peptides are significantly decreased in clpC, clpP, and clpX mutant cells. Electron-dense aggregates, most likely due to the ac...

  11. 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.

  12. Fungal phytases: characteristics and amelioration of nutritional quality and growth of non-ruminants.

    Science.gov (United States)

    Singh, B; Satyanarayana, T

    2015-08-01

    Fungal phytases are histidine acid phosphatases, a subclass of acid phosphatases, which catalyse the hydrolysis of phytic acid resulting in the release of phosphate moieties and thus mitigate its antinutritional properties. The supplementation of feed with phytases increases the bioavailability of phosphorus and minerals in non-ruminant animals and reduces the phosphorus pollution due to phosphorus excretion in the areas of intensive livestock production. Although phytases are reported in plants, animals and micro-organisms, fungal sources are used extensively for the production of phytases on a commercial scale. Phytases have been produced by fungi in both solid-state fermentation (SSF) and submerged fermentation (SmF). The fungal phytases are high molecular weight proteins ranging from 35 to 500 kDa. They are optimally active within pH and temperature ranges between 4.5 and 6.0, and 45 and 70 °C respectively. Phytate degradation leads to amelioration in the nutritional status of foods and feeds by improving the availability of minerals, phosphorus and proteins in non-ruminant animals and human beings and thus mitigates the environmental phosphorus pollution. Our article focuses on the role of fungal phytases in improving nutritional value of foods and feeds with concomitant increase in growth of non-ruminant animals and mitigating environmental phosphorus pollution. Journal of Animal Physiology and Animal Nutrition © 2014 Blackwell Verlag GmbH.

  13. 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.

  14. Effects of feeding lauric acid or coconut oil on ruminal protozoa numbers, fermentation pattern, digestion, omasal nutrient flow, and milk production in dairy cows.

    Science.gov (United States)

    Faciola, A P; Broderick, G A

    2014-01-01

    The objectives of this study were to evaluate the feeding of coconut oil (CO), in which lauric acid (La) comprises about 50% of the fatty acid composition, as a practical rumen protozoa (RP) suppressing agent, to assess whether the source of La affects ruminal fermentation and animal performance and to test whether suppressing RP improves N utilization, nutrient digestion, nutrient flow at the omasal canal, and milk production. Fifteen multiparous Holstein cows (3 fitted with ruminal cannulas) and 15 primiparous Holstein cows (3 fitted with ruminal cannulas) were used in a replicated 3×3 Latin square experiment with 14d of adaptation and 14d of sample collection. Diets were fed as total mixed ration and contained (dry matter basis) 10% corn silage, 50% alfalfa silage, and 40% concentrate. The control diet contained 3% (dry matter basis) calcium soaps of palm oil fatty acids (Megalac, Church & Dwight Co. Inc., Princeton, NJ) as a ruminally inert fat source and had no added La or CO. Diets with La and CO were formulated to contain equal amounts of La (1.3%, dry matter basis). Dry matter intake was not affected by treatment. Both CO and La reduced RP numbers by about 40%. Lauric acid reduced yield of milk and milk components; however, CO did not affect yield of milk and yields of milk components. Both La and CO caused small reductions in total VFA concentration; CO increased molar proportion of ruminal propionate, reduced ruminal ammonia and branched-chain volatile fatty acids, suggesting reduced protein degradation, and reduced milk urea N and blood urea N concentrations, suggesting improved protein efficiency. Lauric acid reduced total-tract apparent digestibility of neutral detergent fiber and acid detergent fiber as well as ruminal apparent digestibility of neutral detergent fiber and acid detergent fiber as measured at the omasal canal; however, CO did not alter fiber digestion. Microbial protein flow at the omasal canal, as well as the flow of N fractions at

  15. Aditivos químicos ou biológicos na ensilagem de cana-de-açúcar: 2. parâmetros ruminais e degradabilidade da matéria seca e das frações fibrosas Chemical and biological additives in sugar cane silages: 2. ruminal parameters and DM and fiber degradabilities

    Directory of Open Access Journals (Sweden)

    Patrick Schmidt

    2007-10-01

    Full Text Available Objetivou-se avaliar o efeito da inclusão de aditivos na ensilagem de cana-de-açúcar (Saccharum officinarum L. sobre a degradação de MS e de componentes da parede celular e sobre os parâmetros de fermentação ruminal em bovinos alimentados com dietas contendo essas silagens. Utilizaram-se cinco novilhos da raça Nelore providos de cânula ruminal, alocados em delineamento quadrado latino 5 ´ 5 e alimentados com dietas com 65% de volumoso (% MS. Foram avaliadas cinco silagens (base úmida: controle - cana-de-açúcar, sem aditivos; uréia - cana-de-açúcar + 0,5% ureia; benzoato - cana-de-açúcar + 0,1% de benzoato de sódio; LP - cana-de-açúcar inoculada com Lactobacillus plantarum (1 ´ 10(6 ufc/g MV; LB - cana-de-açúcar inoculada com L. buchneri (3,6 ´ 10(5 ufc/g forragem. A forragem foi armazenada em silos do tipo poço por 90 dias antes do fornecimento aos animais. Os parâmetros ruminais foram afetados de forma moderada pelas silagens e tiveram forte efeito do horário de coleta de amostras. As concentrações molares médias dos ácidos acético, propiônico e butírico foram de 60,9; 19,3 e 10,2 mM, respectivamente. O ambiente ruminal proporcionado por dietas formuladas com silagens de cana-de-açúcar foi satisfatório e similar ao tradicionalmente observado em dietas contendo cana. O uso de aditivos na ensilagem influenciou, de forma não-significativa, a degradabilidade ruminal da MS e da MO, mas não alterou a degradabilidade ruminal da fração fibrosa. Os aditivos aplicados à cana-de-açúcar resultaram em pequenas alterações na maior parte das variáveis avaliadas. Apesar de a degradabilidade ruminal das silagens ter sido pouco afetada pelo uso de aditivos, os valores observados foram próximos aos observados para a cana-de-açúcar in natura.The objective was to evaluate the addition of additives in the ensiling of sugar cane (Saccharum officinarum L. on the degradation of DM and components of cellular wall and

  16. 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.

  17. Vermicomposting: Tool for Sustainable Ruminant Manure Management

    Directory of Open Access Journals (Sweden)

    A. Nasiru

    2013-01-01

    Full Text Available Ruminants are important sources of meat and milk. Their production is associated with manure excretion. Estimates of over 3,900,000 million metric tonnes of manure are produced daily from ruminants worldwide. Storage and spread of this waste on land pose health risks and environmental problems. Efficient and sustainable way of handling ruminant manure is required. Composting and vermicomposting are considered two of the best techniques for solid biomass waste management. This paper presents vermicomposting as an effective tool for ruminant manure management. Vermicomposting is a mesophilic biooxidation and stabilisation process of organic materials that involves the joint action of earthworm and microorganism. Compared with composting, vermicomposting has higher rate of stabilisation and it is greatly modifying its physical and biochemical properties, with low C : N ratio and homogenous end product. It is also costeffective and ecofriendly waste management. Due to its innate biological, biochemical and physicochemical properties, vermicomposting can be used to promote sustainable ruminant manure management. Vermicomposts are excellent sources of biofertiliser and their addition improves the physiochemical and biological properties of agricultural soils. In addition, earthworms from the vermicomposting can be used as source of protein to fishes and monogastric animals. Vermicompost can also be used as raw materials for bioindustries.

  18. 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

  19. In vitro protein degradation of 38 sainfoin accessions and its relationship to tannin content by different assays.

    Science.gov (United States)

    Lorenz, Martin M; Hayot Carbonero, Christine; Smith, Lydia; Udén, Peter

    2012-05-23

    This study compared 38 sainfoin and 2 Lotus accessions to their respective tannin contents, N buffer solubility, and in vitro protein degradation. Tannin contents were measured by a protein precipitation method using either bovine serum albumin or Rubisco and by the colorimetric HCl/butanol method. Precipitation of bovine serum albumin and Rubisco was highly correlated (R(2) = 0.939). Correlations between the protein precipitation variants and the HCl/butanol method were relatively low (R(2) < 0.6). Protein degradation was measured at 4 h of incubation in an inhibited in vitro system and could not be explained by any of the tannin assays (R(2) < 0.03) and only partially by N buffer solubility (R(2) ≤ 0.433). Decisive factors other than the quantity of tannins or their ability to precipitate proteins must be considered. Resistance of soluble protein toward degradation can possibly be caused by tannin protein binding.

  20. Nitrogen fractionation of certain conventional- and lesser-known by-products for ruminants

    Directory of Open Access Journals (Sweden)

    M.S. Mahesh

    2017-06-01

    Full Text Available Dietary proteins for ruminants are fractionated according to solubility, degradability and digestibility. In the present experiment, 11 vegetable protein meals and cakes used in ruminant nutrition were included with a main focus on determining various nitrogen (N fractions in vitro. Total N (N × 6.25 content varied from 22.98% (mahua cake to 65.16% (maize gluten meal, respectively. Guar meal korma contained the lowest and rice gluten meal had the highest acid detergent insoluble nitrogen (ADIN; N × 6.25. Borate-phosphate insoluble N (BIN, N × 6.25 and Streptomyces griseus protease insoluble N (PIN; N × 6.25 were higher (P < 0.01 in maize gluten meal than in other feeds, whereas groundnut cake and sunflower cake had lower (P < 0.01 BIN, and PIN, respectively. Available N, calculated with the assumption that ADIN is indigestible, was maximum in guar meal korma and minimum in rice gluten meal. Furthermore, rapid and slowly degradable N (N × 6.25 was found to be higher (P < 0.01 in groundnut cake and coconut cake, respectively. Intestinal digestion of rumen undegradable protein, expressed as percent of PIN, was maximum in guar meal korma and minimum in rice gluten meal. It was concluded that vegetable protein meals differed considerably in N fractions, and therefore, a selective inclusion of particular ingredient is needed to achieve desired level of N fractions to aid precision N rationing for an improved production performance of ruminants.

  1. Estimativa da degradabilidade ruminal de quatro genótipos de sorgo (Sorghum bicolor (L. Moench utilizando a técnica in situ - DOI: 10.4025/actascianimsci.v27i4.1150 Estimation of ruminal degradability of four genotypes of sorghum (Sorghum bicolor (L. Moench using in situ technique - DOI: 10.4025/actascianimsci.v27i4.1150

    Directory of Open Access Journals (Sweden)

    Eloísa de Oliveira Simões Saliba

    2005-03-01

    Full Text Available O experimento foi conduzido com o objetivo de avaliar a degradabilidade in situ da matéria seca (MS, da fibra em detergente neutro (FDN e da fibra em detergente ácido (FDA do material original de quatro genótipos de sorgo (ATF53*9929036; ATF54*9929036; CMSXS217*9929012 e VOLUMAX. Foram utilizadas 4 vacas Holandesas providas de fistula ruminal alimentadas com silagem de sorgo “ad libitun” e dois quilos de concentrado. O delineamento experimental foi o de blocos inteiramente ao acaso, com quatro repetições (animais, em arranjo de parcelas subdivididas. Os genótipos constituíram as parcelas e os tempos de digestão as sub-parcelas. O genótipo VOLUMAX foi o que apresentou a maior degradabilidade efetiva (DE da MS (56,22; 53,35 e 50,90% em relação aos demais em todas as taxas de passagem (2, 5 e 8%/h respectivamente e os genótipos ATF53*992903 e ATF54*9929036 obtiveram a maior DE da FDN (32,17 e 33,47%, respectivamente e FDA (34,81 e 35,50%, respectivamente para uma taxa de passagem de 2%/hThis experiment was carried out to evaluate “in situ” degradability of dry matter, neutral detergent fiber (NDF and acid detergent fiber (ADF of four sorghum genotypes (ATF53*9929036; ATF54*9929036; 217*9929012 and VOLUMAX. Four Holstein cows with ruminal fistula were fed on sorghum silage “ad libitum”, and 2.0 kg of concentrate. The animals were alloted in a randomized block design, with four replicates, in a split plot arrangement. The genotypes were considered the parcels and the incubation time the sub-parcels. The higher effective dry matter degradability were found in VOLUMAX genotype, (56.22; 53.35 and 50.90% for all passage rates, (2; 5 and 8%/h, respectively. The genotypes ATF53*992903 and ATF54*9929036 showed higher NDF effective degradability, 32.17 and 33.47% respectively, and ADF effective degradability, 34.81 and 35.50%, at 2%/h passage rate

  2. Strain-Dependent Effect of Macroautophagy on Abnormally Folded Prion Protein Degradation in Infected Neuronal Cells.

    Directory of Open Access Journals (Sweden)

    Daisuke Ishibashi

    Full Text Available Prion diseases are neurodegenerative disorders caused by the accumulation of abnormal prion protein (PrPSc in the central nervous system. With the aim of elucidating the mechanism underlying the accumulation and degradation of PrPSc, we investigated the role of autophagy in its degradation, using cultured cells stably infected with distinct prion strains. The effects of pharmacological compounds that inhibit or stimulate the cellular signal transduction pathways that mediate autophagy during PrPSc degradation were evaluated. The accumulation of PrPSc in cells persistently infected with the prion strain Fukuoka-1 (FK, derived from a patient with Gerstmann-Sträussler-Scheinker syndrome, was significantly increased in cultures treated with the macroautophagy inhibitor 3-methyladenine (3MA but substantially reduced in those treated with the macroautophagy inducer rapamycin. The decrease in FK-derived PrPSc levels was mediated, at least in part, by the phosphatidylinositol 3-kinase/MEK signalling pathway. By contrast, neither rapamycin nor 3MA had any apparently effect on PrPSc from either the 22L or the Chandler strain, indicating that the degradation of PrPSc in host cells might be strain-dependent.

  3. Metformin induces degradation of mTOR protein in breast cancer cells.

    Science.gov (United States)

    Alalem, Mohamed; Ray, Alpana; Ray, Bimal K

    2016-11-01

    Activation of mTOR is implicated in the development and progression of breast cancer. mTOR inhibition exhibited promising antitumor effects in breast cancer; however, its effect is compromised by several feedback mechanisms. One of such mechanisms is the upregulation of mTOR pathway in breast cancer cells. Despite the established role of mTOR activation in breast cancer, the status of total mTOR protein and its impact on the tumor behavior and response to treatment are poorly understood. Besides, the mechanisms underlying mTOR protein degradation in normal and cancer breast cells are still largely unknown. We and others found that total mTOR protein level is elevated in breast cancer cells compared to their nonmalignant counterparts. We have detected defective proteolysis of mTOR protein in breast cancer cells, which could, at least in part, explain the high level of mTOR protein in these cells. We show that metformin treatment in MCF-7 breast cancer cells induced degradation of mTOR and sequestration of this protein in a perinuclear region. The decrease in mTOR protein level in these cells correlated positively with a concomitant inhibition of proliferation and migration potentials of these cells. These findings provided a novel mechanism for the metformin action in breast cancer treatment. Understanding the proteolytic mechanism responsible for mTOR level in breast cancer may pave the way for improving the efficacy of breast cancer treatment regimens and mitigating drug resistance as well as providing a basis for potential novel therapeutic modalities for breast cancer. © 2016 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.

  4. Calpain-2-mediated PTEN degradation contributes to BDNF-induced stimulation of dendritic protein synthesis

    Science.gov (United States)

    Briz, Victor; Hsu, Yu-Tien; Li, Yi; Lee, Erin; Bi, Xiaoning; Baudry, Michel

    2013-01-01

    Memory consolidation has been suggested to be protein synthesis-dependent. Recent data indicate that BDNF-induced dendritic protein synthesis is a key event in memory formation through activation of the mammalian target of rapamycin (mTOR) pathway. BDNF also activates calpain, a calcium-dependent cysteine protease, which has been shown to play a critical role in learning and memory. This study was therefore directed at testing the hypothesis that calpain activity is required for BDNF-stimulated local protein synthesis, and at identifying the underlying molecular mechanism. In rat hippocampal slices, cortical synaptoneurosomes, and cultured neurons, BDNF-induced mTOR pathway activation and protein translation were blocked by calpain inhibition. BDNF treatment rapidly reduced levels of hamartin and tuberin, negative regulators of mTOR, in a calpain-dependent manner. Treatment of brain homogenates with purified calpain-1 and calpain-2 truncated both proteins. BDNF treatment increased phosphorylation of both Akt and ERK, but only the effect on Akt was blocked by calpain inhibition. Levels of PTEN (phosphatase and tensin homolog deleted on chromosome ten), a phosphatase that inactivates Akt, were decreased following BDNF treatment, and calpain inhibition reversed this effect. Calpain-2 but not calpain-1 treatment of brain homogenates resulted in PTEN degradation. In cultured cortical neurons, knock-down of calpain-2 but not calpain-1 by siRNA completely suppressed the effect of BDNF on mTOR activation. Our results reveal a critical role for calpain-2 in BDNF-induced mTOR signaling and dendritic protein synthesis via PTEN, hamartin and tuberin degradation. This mechanism therefore provides a link between proteolysis and protein synthesis that might contribute to synaptic plasticity. PMID:23467348

  5. Review: Use of human-edible animal feeds by ruminant livestock.

    Science.gov (United States)

    Wilkinson, J M; Lee, M R F

    2017-09-12

    The drive to increase the output of animal product in some sectors of ruminant livestock production has led to greater use of feeds such as cereal grains and soyabean meal that are potentially human-edible. This trend has caused concern since, by so doing, ruminants compete not only with monogastric livestock but also with the human population for a limited global area of cultivatable land on which to produce grain crops. Reasons for using potentially human-edible feeds in ruminant diets include increased total daily energy intake, greater supply of essential amino acids and improved ruminal balance between fermentable energy and degradable protein. Soyabean meal, produced on land that has been in arable cultivation for many years can fulfil a useful role as a supplier of undegraded dietary protein in diets for high-yielding dairy cows. However, in the context of sustaining the production of high-quality foods from livestock to meet the demands of a growing human population, the use of potentially human-edible feed resources by livestock should be restricted to livestock with the highest daily nutrient requirements; that is, potentially human-edible feed inputs should be constrained to meeting requirements for energy and protein and to rectifying imbalances in nutrient supply from pastures and forage crops such as high concentrations of nitrogen (N). There is therefore a role for human-edible feeds in milk production because forage-only systems are associated with relatively low output per head and also low N use efficiency compared with systems with greater reliance on human-edible feeds. Profitability on farm is driven by control of input costs as well as product value and examples are given of low-cost bovine milk and meat production with little or no reliance on potentially human-edible feeds. In beef production, the forage-only systems currently under detailed real-time life-cycle analysis at the North Wyke Farm Platform, can sustain high levels of animal

  6. Hepatitis B Virus X Protein Promotes Degradation of SMC5/6 to Enhance HBV Replication

    Directory of Open Access Journals (Sweden)

    Christopher M. Murphy

    2016-09-01

    Full Text Available The hepatitis B virus (HBV regulatory protein X (HBx activates gene expression from the HBV covalently closed circular DNA (cccDNA genome. Interaction of HBx with the DDB1-CUL4-ROC1 (CRL4 E3 ligase is critical for this function. Using substrate-trapping proteomics, we identified the structural maintenance of chromosomes (SMC complex proteins SMC5 and SMC6 as CRL4HBx substrates. HBx expression and HBV infection degraded the SMC5/6 complex in human hepatocytes in vitro and in humanized mice in vivo. HBx targets SMC5/6 for ubiquitylation by the CRL4HBx E3 ligase and subsequent degradation by the proteasome. Using a minicircle HBV (mcHBV reporter system with HBx-dependent activity, we demonstrate that SMC5/6 knockdown, or inhibition with a dominant-negative SMC6, enhance HBx null mcHBV-Gluc gene expression. Furthermore, SMC5/6 knockdown rescued HBx-deficient HBV replication in human hepatocytes. These results indicate that a primary function of HBx is to degrade SMC5/6, which restricts HBV replication by inhibiting HBV gene expression.

  7. A parasite cysteine protease is key to host protein degradation and iron acquisition.

    Science.gov (United States)

    O'Brien, Theresa C; Mackey, Zachary B; Fetter, Richard D; Choe, Youngchool; O'Donoghue, Anthony J; Zhou, Min; Craik, Charles S; Caffrey, Conor R; McKerrow, James H

    2008-10-24

    Cysteine proteases of the Clan CA (papain) family are the predominant protease group in primitive invertebrates. Cysteine protease inhibitors arrest infection by the protozoan parasite, Trypanosoma brucei. RNA interference studies implicated a cathepsin B-like protease, tbcatB, as a key inhibitor target. Utilizing parasites in which one of the two alleles of tbcatb has been deleted, the key role of this protease in degradation of endocytosed host proteins is delineated. TbcatB deficiency results in a decreased growth rate and dysmorphism of the flagellar pocket and the subjacent endocytic compartment. Western blot and microscopic analysis indicate that deficiency in tbcatB results in accumulation of both host and parasite proteins, including the lysosomal marker p67. A critical function for parasitism is the degradation of host transferrin, which is necessary for iron acquisition. Substrate specificity analysis of recombinant tbcatB revealed the optimal peptide cleavage sequences for the enzyme and these were confirmed experimentally using FRET-based substrates. Degradation of transferrin was validated by SDS-PAGE and the specific cleavage sites identified by N-terminal sequencing. Because even a modest deficiency in tbcatB is lethal for the parasite, tbcatB is a logical target for the development of new anti-trypanosomal chemotherapy.

  8. Organic Zinc as Feed Additive for Ruminants

    Directory of Open Access Journals (Sweden)

    Suprijati

    2013-09-01

    Full Text Available Zinc is an essential micro mineral required by ruminants and is a component of over 300 enzymes which play important role in the metabolisms of carbohydrates, proteins and fats. Recently, the chemical and biotechnology processes have been developed for synthesizing organic Zn. Organic Zn is the product of a chelating process of dissolved Zn anorganic salts with amino acids or hydrolyzed protein. The utilization of organic Zn as feed additive in ruminants diets tends to increase, due to the absorption of Zn in the organic form is more effective than the inorganic form. This paper reviewes the research from comparative studies of inorganic and organic Zn on the growth, production and reproduction in ruminants. The studies found that the supplementation of organic Zn improved nutrient digestibility, growth, feed efficiency, milk production, Zn retention and absorption, production and reproductive performances of ruminants. However, a little variation response was found due to different degrees of chelating on Zn incorporation of amino acids or hydrolyzed protein.

  9. 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.

  10. Particulate matter air pollution disrupts endothelial cell barrier via calpain-mediated tight junction protein degradation.

    Science.gov (United States)

    Wang, Ting; Wang, Lichun; Moreno-Vinasco, Liliana; Lang, Gabriel D; Siegler, Jessica H; Mathew, Biji; Usatyuk, Peter V; Samet, Jonathan M; Geyh, Alison S; Breysse, Patrick N; Natarajan, Viswanathan; Garcia, Joe G N

    2012-08-29

    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. 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. 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. 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. 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.

  11. Insulin-degrading enzyme is exported via an unconventional protein secretion pathway

    Directory of Open Access Journals (Sweden)

    Leissring Malcolm A

    2009-01-01

    Full Text Available Abstract Insulin-degrading enzyme (IDE is a ubiquitously expressed zinc-metalloprotease that degrades several pathophysiologically significant extracellular substrates, including insulin and the amyloid β-protein (Aβ, and accumulating evidence suggests that IDE dysfunction may be operative in both type 2 diabetes mellitus and Alzheimer disease (AD. Although IDE is well known to be secreted by a variety of cell types, the underlying trafficking pathway(s remain poorly understood. To address this topic, we investigated the effects of known inhibitors or stimulators of protein secretion on the secretion of IDE from murine hepatocytes and HeLa cells. IDE secretion was found to be unaffected by the classical secretion inhibitors brefeldin A (BFA, monensin, or nocodazole, treatments that readily inhibited the secretion of α1-antitrypsin (AAT overexpressed in the same cells. Using a novel cell-based Aβ-degradation assay, we show further that IDE secretion was similarly unaffected by multiple stimulators of protein secretion, including glyburide and 3'-O-(4-benzoylbenzoyl-ATP (Bz-ATP. The calcium ionophore, A23187, increased extracellular IDE activity, but only under conditions that also elicited cytotoxicity. Our results provide the first biochemical evidence that IDE export is not dependent upon the classical secretion pathway, thereby identifying IDE as a novel member of the select class of unconventionally secreted proteins. Further elucidation of the mechanisms underlying IDE secretion, which would be facilitated by the assays described herein, promises to uncover processes that might be defective in disease or manipulated for therapeutic benefit.

  12. Legume-Cereal Intercropping Improves Forage Yield, Quality and Degradability.

    Science.gov (United States)

    Zhang, Jie; Yin, Binjie; Xie, Yuhuai; Li, Jing; Yang, Zaibin; Zhang, Guiguo

    2015-01-01

    Intercropping legume with cereal is an extensively applied planting pattern in crop cultivation. However, forage potential and the degradability of harvested mixtures from intercropping system remain unclear. To investigate the feasibility of applying an intercropping system as a forage supply source to ruminants, two consecutive experiments (experiments 1 and 2) involving a field cultivation trial and a subsequent in vivo degradable experiment were conducted to determine the forage production performance and the ruminally degradable characteristics of a harvested mixture from an alfalfa/corn-rye intercropping system. In experiment 1, the intercropping system was established by alternating alfalfa and corn or rye with a row ratio of 5:2. Dry matter (DM) and nutrient yields were determined. In experiment 2, forages harvested from the different treatments were used as feedstuff to identify nutrient degradation kinetics and distribution of components between the rapidly degradable (a), potentially degradable (b) and the degradation rate constant (c) of 'b' fraction by in sacco method in Small-Tail Han wether Sheep. The intercropping system of alfalfa and corn-rye provided higher forage production performance with net increases of 9.52% and 34.81% in DM yield, 42.13% and 16.74% in crude protein (CP) yield, 25.94% and 69.99% in degradable DM yield, and 16.96% and 5.50% in degradable CP yield than rotation and alfalfa sole cropping systems, respectively. In addition, the harvest mixture from intercropping system also had greater 'a' fraction, 'b' fraction, 'c' values, and effective degradability (E value) of DM and CP than corn or rye hay harvested from rotation system. After 48-h exposure to rumen microbes, intercropping harvest materials were degraded to a higher extent than separately degraded crop stems from the sole system as indicated by visual microscopic examination with more tissues disappeared. Thus, the intercropping of alfalfa and corn-rye exhibited a greater

  13. 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

  14. Recent advances in modeling nutrient utilization in ruminants.

    Science.gov (United States)

    Kebreab, E; Dijkstra, J; Bannink, A; France, J

    2009-04-01

    Mathematical modeling techniques have been applied to study various aspects of the ruminant, such as rumen function, postabsorptive metabolism, and product composition. This review focuses on advances made in modeling rumen fermentation and its associated rumen disorders, and energy and nutrient utilization and excretion with respect to environmental issues. Accurate prediction of fermentation stoichiometry has an impact on estimating the type of energy-yielding substrate available to the animal, and the ratio of lipogenic to glucogenic VFA is an important determinant of methanogenesis. Recent advances in modeling VFA stoichiometry offer ways for dietary manipulation to shift the fermentation in favor of glucogenic VFA. Increasing energy to the animal by supplementing with starch can lead to health problems such as subacute rumen acidosis caused by rumen pH depression. Mathematical models have been developed to describe changes in rumen pH and rumen fermentation. Models that relate rumen temperature to rumen pH have also been developed and have the potential to aid in the diagnosis of subacute rumen acidosis. The effect of pH has been studied mechanistically, and in such models, fractional passage rate has a large impact on substrate degradation and microbial efficiency in the rumen and should be an important theme in future studies. The efficiency with which energy is utilized by ruminants has been updated in recent studies. Mechanistic models of N utilization indicate that reducing dietary protein concentration, matching protein degradability to the microbial requirement, and increasing the energy status of the animal will reduce the output of N as waste. Recent mechanistic P models calculate the P requirement by taking into account P recycled through saliva and endogenous losses. Mechanistic P models suggest reducing current P amounts for lactating dairy cattle to at least 0.35% P in the diet, with a potential reduction of up to 1.3 kt/yr. A model that

  15. Rumination: Relationships with Physical Health

    OpenAIRE

    Sansone, Randy A.; Sansone, Lori A.

    2012-01-01

    Rumination is a form of perserverative cognition that focuses on negative content, generally past and present, and results in emotional distress. Initial studies of rumination emerged in the psychological literature, particularly with regard to studies examining specific facets of rumination (e.g., positive vs. negative rumination, brooding vs. self-reflection, relationships with catastrophic thinking, role of impaired disengagement, state vs. trait features) as well as the presence of rumina...

  16. Ectomycorrhizal Fungal Protein Degradation Ability Predicted by Soil Organic Nitrogen Availability.

    Science.gov (United States)

    Rineau, Francois; Stas, Jelle; Nguyen, Nhu H; Kuyper, Thomas W; Carleer, Robert; Vangronsveld, Jaco; Colpaert, Jan V; Kennedy, Peter G

    2015-12-18

    In temperate and boreal forest ecosystems, nitrogen (N) limitation of tree metabolism is alleviated by ectomycorrhizal (ECM) fungi. As forest soils age, the primary source of N in soil switches from inorganic (NH4 (+) and NO3 (-)) to organic (mostly proteins). It has been hypothesized that ECM fungi adapt to the most common N source in their environment, which implies that fungi growing in older forests would have greater protein degradation abilities. Moreover, recent results for a model ECM fungal species suggest that organic N uptake requires a glucose supply. To test the generality of these hypotheses, we screened 55 strains of 13 Suillus species with different ecological preferences for their in vitro protein degradation abilities. Suillus species preferentially occurring in mature forests, where soil contains more organic matter, had significantly higher protease activity than those from young forests with low-organic-matter soils or species indifferent to forest age. Within species, the protease activities of ecotypes from soils with high or low soil organic N content did not differ significantly, suggesting resource partitioning between mineral and organic soil layers. The secreted protease mixtures were strongly dominated by aspartic peptidases. Glucose addition had variable effects on secreted protease activity; in some species, it triggered activity, but in others, activity was repressed at high concentrations. Collectively, our results indicate that protease activity, a key ectomycorrhizal functional trait, is positively related to environmental N source availability but is also influenced by additional factors, such as carbon availability. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  17. 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...

  18. 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.

  19. [Local anaesthesia in ruminants].

    Science.gov (United States)

    Nuss, Karl; Schwarz, Andrea; Ringer, Simone

    2017-06-20

    The use of local anaesthesia in ruminants allows many surgical procedures to be conducted free of pain, efficiently and inexpensively in the field. Local anaesthesia combined with sedation and immobilisation of the animal can replace general anaesthesia for many procedures (e. g. castration, claw amputation). The level of difficulty differs among various local anaesthetic techniques: local infiltration of tissue or anaesthesia of the cornual nerve are easily performed, whereas local anaesthesia of the eye, regional anaesthesia in limbs or anaesthesia for umbilical surgery are more difficult to carry out. This article presents an illustrated overview of the most common local anaesthetic procedures in cattle as well as in small ruminants and serves as a practical guide for veterinarians in the field. In principle, these techniques can likewise be applied in other ruminants or artiodactyls.

  20. Interpretation bias characterizes trait rumination.

    Science.gov (United States)

    Mor, Nilly; Hertel, Paula; Ngo, Thuy Anh; Shachar, Tal; Redak, Shimrit

    2014-03-01

    Rumination, a maladaptive cognitive style of responding to negative mood, is thought to be maintained by a variety of cognitive biases. However, it is unknown whether rumination is characterized by interpretation biases. Two experiments examined the link between rumination and interpretation biases, revealed in lexical-decision tasks (LDT). A homograph with both benign and ruminative or otherwise negative meaning was presented on each trial and followed by a letter string, to which participants responded by judging whether it was a word or a non-word. Letter strings were non-words or words related or unrelated to one meaning of the homograph. In both experiments, faster latencies to respond to targets related to the ruminative meaning of the homographs were produced by students with higher scores on self-report measures of rumination. Moreover, these biases were associated with both brooding, the maladaptive form of rumination, and reflection, the more adaptive component. No measure of rumination was significantly correlated with general biases toward negative meaning (Experiment 1) or with threatening interpretations of homographs (Experiment 2). The paucity of available rumination-related homographs dictated the use of non-fully randomized stimuli presentation (Experiment 1) or the use of only one set of the meanings associated with the homographs (Experiment 2). Rumination is associated with a tendency to interpret ambiguous information in a rumination-consistent manner. This tendency may exacerbate ruminative thinking and can possibly be a target for future intervention. Copyright © 2013 Elsevier Ltd. All rights reserved.

  1. Ribosomal Protein Mutations Result in Constitutive p53 Protein Degradation through Impairment of the AKT Pathway

    NARCIS (Netherlands)

    Antunes, Ana T.; Goos, Yvonne J.; Pereboom, Tamara C.; Hermkens, Dorien; Wlodarski, Marcin W.; Da Costa, Lydie; MacInnes, Alyson W.

    Mutations in ribosomal protein (RP) genes can result in the loss of erythrocyte progenitor cells and cause severe anemia. This is seen in patients with Diamond-Blackfan anemia (DBA), a pure red cell aplasia and bone marrow failure syndrome that is almost exclusively linked to RP gene

  2. Staphylococcus epidermidis Esp degrades specific proteins associated with Staphylococcus aureus biofilm formation and host-pathogen interaction.

    Science.gov (United States)

    Sugimoto, Shinya; Iwamoto, Takeo; Takada, Koji; Okuda, Ken-Ichi; Tajima, Akiko; Iwase, Tadayuki; Mizunoe, Yoshimitsu

    2013-04-01

    Staphylococcus aureus exhibits a strong capacity to attach to abiotic or biotic surfaces and form biofilms, which lead to chronic infections. We have recently shown that Esp, a serine protease secreted by commensal Staphylococcus epidermidis, disassembles preformed biofilms of S. aureus and inhibits its colonization. Esp was expected to degrade protein determinants of the adhesive and cohesive strength of S. aureus biofilms. The aim of this study was to elucidate the substrate specificity and target proteins of Esp and thereby determine the mechanism by which Esp disassembles S. aureus biofilms. We used a mutant Esp protein (Esp(S235A)) with defective proteolytic activity; this protein did not disassemble the biofilm formed by a clinically isolated methicillin-resistant S. aureus (MRSA) strain, thereby indicating that the proteolytic activity of Esp is essential for biofilm disassembly. Esp degraded specific proteins in the biofilm matrix and cell wall fractions, in contrast to proteinase K, which is frequently used for testing biofilm robustness and showed no preference for proteolysis. Proteomic and immunological analyses showed that Esp degrades at least 75 proteins, including 11 biofilm formation- and colonization-associated proteins, such as the extracellular adherence protein, the extracellular matrix protein-binding protein, fibronectin-binding protein A, and protein A. In addition, Esp selectively degraded several human receptor proteins of S. aureus (e.g., fibronectin, fibrinogen, and vitronectin) that are involved in its colonization or infection. These results suggest that Esp inhibits S. aureus colonization and biofilm formation by degrading specific proteins that are crucial for biofilm construction and host-pathogen interaction.

  3. Rumen Degradability and Small Intestinal Digestibility of the Amino Acids in Four Protein Supplements

    Science.gov (United States)

    Wang, Y.; Jin, L.; Wen, Q. N.; Kopparapu, N. K.; Liu, J.; Liu, X. L.; Zhang, Y. G.

    2016-01-01

    The supplementation of livestock feed with animal protein is a present cause for public concern, and plant protein shortages have become increasingly prominent in China. This conflict may be resolved by fully utilizing currently available sources of plant protein. We estimated the rumen degradability and the small intestinal digestibility of the amino acids (AA) in rapeseed meal (RSM), soybean meal (SBM), sunflower seed meal (SFM) and sesame meal (SSM) using the mobile nylon bag method to determine the absorbable AA content of these protein supplements as a guide towards dietary formulations for the dairy industry. Overall, this study aimed to utilize protein supplements effectively to guide dietary formulations to increase milk yield and save plant protein resources. To this end, we studied four cows with a permanent rumen fistula and duodenal T-shape fistula in a 4×4 Latin square experimental design. The results showed that the total small intestine absorbable amino acids and small intestine absorbable essential amino acids were higher in the SBM (26.34% and 13.11% dry matter [DM], respectively) than in the SFM (13.97% and 6.89% DM, respectively). The small intestine absorbable Lys contents of the SFM, SSM, RSM and SBM were 0.86%, 0.88%, 1.43%, and 2.12% (DM basis), respectively, and the absorbable Met contents of these meals were 0.28%, 1.03%, 0.52%, and 0.47% (DM basis), respectively. Among the examined food sources, the milk protein score of the SBM (0.181) was highest followed by those of the RSM (0.136), SSM (0.108) and SFM (0.106). The absorbable amino acid contents of the protein supplements accurately reflected protein availability, which is an important indicator of the balance of feed formulation. Therefore, a database detailing the absorbable AA should be established. PMID:26732449

  4. Wet degradation of keratin proteins: linking amino acid, elemental and isotopic composition.

    Science.gov (United States)

    von Holstein, I C C; Penkman, K E H; Peacock, E E; Collins, M J

    2014-10-15

    Archaeological keratin samples are increasingly the subject of palaeodietary, provenancing and dating studies. Keratin samples from wet archaeological contexts are microbiologically and chemically degraded, causing differential diagenesis of protein structures in hair fibres. The effects of these processes on the analytical parameters of interest are currently unknown. This study examined the impact of degradation of wool fibres on isotopic (δ(13)C, δ(15)N, un-exchangeable δ(2)H and δ(18)O values) composition. It compared two models of archaeological protein degradation in wet burial environments: (1) short term (up to 8 years) experimental burial in three contrasting soil environments; and (2) laboratory wet conditions, in which elevated temperature (80 °C, 110 °C, and 140 °C) and pressure simulated longer exposure. Elemental and amino acid (AA) composition were also measured. In experimentally soil-buried samples, AA, elemental and isotopic composition changes were small, despite extensive macroscopic alteration. Isothermally heated samples showed preferential loss of hydrophilic AAs (Asx, Glx, Ser, Gly) from wool residues, with depletion in (2)H and (18)O at higher temperatures (up to -73‰ change in δ(2)H and -2.6‰ in δ(18)O values). The δ(13)C and δ(15)N values showed little change except in densely pigmented samples at low temperatures only. Samples dyed with madder/alum were better preserved than undyed samples. Diagenesis in experimentally soil-buried wool textiles was consistent with microbiological, non-protein-selective activity, in contrast to highly AA-selective hydrolytic behaviour under laboratory wet conditions. Changes in δ(2)H and δ(18)O values were correlated with degree of AA change, but the δ(13)C and δ(15)N values were not. The results contribute to a baseline for interpreting analytical data from archaeological hair samples preserved by burial in wet environments. Copyright © 2014 John Wiley & Sons, Ltd.

  5. DHA Inhibits Protein Degradation More Efficiently than EPA by Regulating the PPARγ/NFκB Pathway in C2C12 Myotubes

    Directory of Open Access Journals (Sweden)

    Yue Wang

    2013-01-01

    Full Text Available This study was conducted to evaluate the mechanism by which n-3 PUFA regulated the protein degradation in C2C12 myotubes. Compared with the BSA control, EPA at concentrations from 400 to 600 µM decreased total protein degradation (P0.05. Interestingly, EPA and DHA both still decreased the total protein degradation, although PPARγ knockdown attenuated the suppressive effects of EPA and DHA on the total protein degradation (P<0.01. These results revealed that DHA inhibits protein degradation more efficiently than EPA by regulating the PPARγ/NF-κB pathway in C2C12 myotubes.

  6. 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.

  7. BDNF Induces Striatal-Enriched Protein Tyrosine Phosphatase 61 Degradation Through the Proteasome.

    Science.gov (United States)

    Saavedra, Ana; Puigdellívol, Mar; Tyebji, Shiraz; Kurup, Pradeep; Xu, Jian; Ginés, Silvia; Alberch, Jordi; Lombroso, Paul J; Pérez-Navarro, Esther

    2016-08-01

    Brain-derived neurotrophic factor (BDNF) promotes synaptic strengthening through the regulation of kinase and phosphatase activity. Conversely, striatal-enriched protein tyrosine phosphatase (STEP) opposes synaptic strengthening through inactivation or internalization of signaling molecules. Here, we investigated whether BDNF regulates STEP levels/activity. BDNF induced a reduction of STEP61 levels in primary cortical neurons, an effect that was prevented by inhibition of tyrosine kinases, phospholipase C gamma, or the ubiquitin-proteasome system (UPS). The levels of pGluN2B(Tyr1472) and pERK1/2(Thr202/Tyr204), two STEP substrates, increased in BDNF-treated cultures, and blockade of the UPS prevented STEP61 degradation and reduced BDNF-induced GluN2B and ERK1/2 phosphorylation. Moreover, brief or sustained cell depolarization reduced STEP61 levels in cortical neurons by different mechanisms. BDNF also promoted UPS-mediated STEP61 degradation in cultured striatal and hippocampal neurons. In contrast, nerve growth factor and neurotrophin-3 had no effect on STEP61 levels. Our results thus indicate that STEP61 degradation is an important event in BDNF-mediated effects.

  8. Estimation of indigestible NDF in feedstuffs for ruminants

    DEFF Research Database (Denmark)

    Krämer, Monika; Weisbjerg, Martin Riis; Lund, Peter

    2010-01-01

    Intrinsic properties of plant cell walls determine the digestibility of ruminant diets, as they establish the maximum degree of the rate and extent of cell wall digestion in ruminants. The determination of INDF is important for the estimation of potentially digestible NDF (DNDF), and has been shown...... to be a suitable parameter in prediction models of energy and protein values in feedstuffs for ruminants, as the NorFor system. Therefore, there is a need to develop laboratory methods, applicable in practice, that determine the INDF content in feedstuffs. The present paper aims at presenting correlations...

  9. A toolkit for GFP-mediated tissue-specific protein degradation in C. elegans.

    Science.gov (United States)

    Wang, Shaohe; Tang, Ngang Heok; Lara-Gonzalez, Pablo; Zhao, Zhiling; Cheerambathur, Dhanya K; Prevo, Bram; Chisholm, Andrew D; Desai, Arshad; Oegema, Karen

    2017-07-15

    Proteins that are essential for embryo production, cell division and early embryonic events are frequently reused later in embryogenesis, during organismal development or in the adult. Examining protein function across these different biological contexts requires tissue-specific perturbation. Here, we describe a method that uses expression of a fusion between a GFP-targeting nanobody and a SOCS-box containing ubiquitin ligase adaptor to target GFP-tagged proteins for degradation. When combined with endogenous locus GFP tagging by CRISPR-Cas9 or with rescue of a null mutant with a GFP fusion, this approach enables routine and efficient tissue-specific protein ablation. We show that this approach works in multiple tissues - the epidermis, intestine, body wall muscle, ciliated sensory neurons and touch receptor neurons - where it recapitulates expected loss-of-function mutant phenotypes. The transgene toolkit and the strain set described here will complement existing approaches to enable routine analysis of the tissue-specific roles of C. elegans proteins. © 2017. Published by The Company of Biologists Ltd.

  10. Protein Phosphatase Methyl-Esterase PME-1 Protects Protein Phosphatase 2A from Ubiquitin/Proteasome Degradation.

    Directory of Open Access Journals (Sweden)

    Ryotaro Yabe

    Full Text Available Protein phosphatase 2A (PP2A is a conserved essential enzyme that is implicated as a tumor suppressor based on its central role in phosphorylation-dependent signaling pathways. Protein phosphatase methyl esterase (PME-1 catalyzes specifically the demethylation of the C-terminal Leu309 residue of PP2A catalytic subunit (PP2Ac. It has been shown that PME-1 affects the activity of PP2A by demethylating PP2Ac, but also by directly binding to the phosphatase active site, suggesting loss of PME-1 in cells would enhance PP2A activity. However, here we show that PME-1 knockout mouse embryonic fibroblasts (MEFs exhibit lower PP2A activity than wild type MEFs. Loss of PME-1 enhanced poly-ubiquitination of PP2Ac and shortened the half-life of PP2Ac protein resulting in reduced PP2Ac levels. Chemical inhibition of PME-1 and rescue experiments with wild type and mutated PME-1 revealed methyl-esterase activity was necessary to maintain PP2Ac protein levels. Our data demonstrate that PME-1 methyl-esterase activity protects PP2Ac from ubiquitin/proteasome degradation.

  11. Limitations to ruminal absorption of volatile fatty acids in lactating dairy cows

    DEFF Research Database (Denmark)

    Storm, Adam Christian

    bodies are released for the benefit of the host animal. The main end product of ruminal fermentation is volatile fatty acids (VFA), which provide the cow with the majority of energetic precursors for metabolic processes. Ruminal VFA are therefore, quantitatively the most important nutrient in cattle...... to ruminal VFA absorption other than the ruminal production rates of VFA in lactating dairy cows, under the hypothesis that ruminal VFA absorption can be limited in three ways: a) VFA absorption can be limited by the extent of intraruminal VFA equilibration of fluid and associated VFA, from the site......The symbiotic relationship between ruminants and the microbial inhabitants of the rumen constitutes a unique feature of the ruminant digestive system. Through the microbial utilization of feed carbohydrates and protein in the rumen, substantial amounts of fermentation products and microbial cell...

  12. The key to unlock the Hsp100/Clp protein degradation machines of Mycobacterium.

    Science.gov (United States)

    Molière, Noël; Turgay, Kürşad

    2014-08-01

    Hsp100/Clp protease complexes are molecular machines important for cellular protein homeostasis and are concurrently embedded in the control of various signal transduction networks by regulatory proteolysis. In Mycobacteria, the genes encoding the components of these Hsp100/Clp protease complexes are essential for growth and were identified as targets for antibiotics, with a new antimicrobial mechanism, that are active on slow growing or even dormant cells. Schmitz and Sauer (2014) report the biochemical characterization of mycobacterial Hsp100/Clp protease complexes actively degrading folded substrate proteins. Their results suggest an unusual activation mechanism for this protease complex and will set the stage for further mechanistic studies of antibiotics acting on this new cellular target. © 2014 John Wiley & Sons Ltd.

  13. RUMINANT NUTRITION SYMPOSIUM: Use of genomics and transcriptomics to identify strategies to lower ruminal methanogenesis.

    Science.gov (United States)

    McAllister, T A; Meale, S J; Valle, E; Guan, L L; Zhou, M; Kelly, W J; Henderson, G; Attwood, G T; Janssen, P H

    2015-04-01

    Globally, methane (CH4) emissions account for 40% to 45% of greenhouse gas emissions from ruminant livestock, with over 90% of these emissions arising from enteric fermentation. Reduction of carbon dioxide to CH4 is critical for efficient ruminal fermentation because it prevents the accumulation of reducing equivalents in the rumen. Methanogens exist in a symbiotic relationship with rumen protozoa and fungi and within biofilms associated with feed and the rumen wall. Genomics and transcriptomics are playing an increasingly important role in defining the ecology of ruminal methanogenesis and identifying avenues for its mitigation. Metagenomic approaches have provided information on changes in abundances as well as the species composition of the methanogen community among ruminants that vary naturally in their CH4 emissions, their feed efficiency, and their response to CH4 mitigators. Sequencing the genomes of rumen methanogens has provided insight into surface proteins that may prove useful in the development of vaccines and has allowed assembly of biochemical pathways for use in chemogenomic approaches to lowering ruminal CH4 emissions. Metagenomics and metatranscriptomic analysis of entire rumen microbial communities are providing new perspectives on how methanogens interact with other members of this ecosystem and how these relationships may be altered to reduce methanogenesis. Identification of community members that produce antimethanogen agents that either inhibit or kill methanogens could lead to the identification of new mitigation approaches. Discovery of a lytic archaeophage that specifically lyses methanogens is 1 such example. Efforts in using genomic data to alter methanogenesis have been hampered by a lack of sequence information that is specific to the microbial community of the rumen. Programs such as Hungate1000 and the Global Rumen Census are increasing the breadth and depth of our understanding of global ruminal microbial communities, steps that

  14. Transient degradation of NF-κB proteins in macrophages after interaction with mast cell granules

    Directory of Open Access Journals (Sweden)

    Noriko Ito

    1998-01-01

    Full Text Available The exposure of the macrophage cell line, J774 to mast cell granules (MCG led to the form ation of altered nuclear transcription factor proteins (NFκBx, which had faster electrophoretic mobility than the p50 homodimer of NF-κB, but retained comparable DNA binding capacity. Antibodies to N-terminal peptides of p50, p52, p65 or c-Rel supershifted only a fraction of NF-κBx. Western blot analyses revealed that nuclear p65 and c-Rel were progressively degraded after exposure to MCG, whereas nuclear p50 appeared to be unaffected. In contrast, cytoplasmic p50, p65, c-Rel as well as IkBα remained intact after MCG treatment, although p52 was clearly degraded. In comparison to J774 cells, incubation of m ouse peritoneal macrophages with MCG resulted in more extensive alterations to NF-κB proteins. The alterations in NF-κB proteins did not affect the expression of inducible nitric oxide synthase (iNOS or TNF-α mRNA in J774 cells. These data indicate that exposure of J774 cells to MCG leads to generation of altered nuclear p52, p65 and c-Rel, which retain intact N-terminal peptides, specific oligonucleotide binding and transactivating activity. On the other hand, in peritoneal macrophages, MCG induce more extensive modifications to NF-κB proteins with associated inhibition of iNOS or TNF-α mRNA expression.

  15. Hepatitis C virus induces E6AP-dependent degradation of the retinoblastoma protein.

    Directory of Open Access Journals (Sweden)

    Tsubasa Munakata

    2007-09-01

    Full Text Available Hepatitis C virus (HCV is a positive-strand RNA virus that frequently causes persistent infections and is uniquely associated with the development of hepatocellular carcinoma. While the mechanism(s by which the virus promotes cancer are poorly defined, previous studies indicate that the HCV RNA-dependent RNA polymerase, nonstructural protein 5B (NS5B, forms a complex with the retinoblastoma tumor suppressor protein (pRb, targeting it for degradation, activating E2F-responsive promoters, and stimulating cellular proliferation. Here, we describe the mechanism underlying pRb regulation by HCV and its relevance to HCV infection. We show that the abundance of pRb is strongly downregulated, and its normal nuclear localization altered to include a major cytoplasmic component, following infection of cultured hepatoma cells with either genotype 1a or 2a HCV. We further demonstrate that this is due to NS5B-dependent ubiquitination of pRb and its subsequent degradation via the proteasome. The NS5B-dependent ubiquitination of pRb requires the ubiquitin ligase activity of E6-associated protein (E6AP, as pRb abundance was restored by siRNA knockdown of E6AP or overexpression of a dominant-negative E6AP mutant in cells containing HCV RNA replicons. E6AP also forms a complex with pRb in an NS5B-dependent manner. These findings suggest a novel mechanism for the regulation of pRb in which the HCV NS5B protein traps pRb in the cytoplasm, and subsequently recruits E6AP to this complex in a process that leads to the ubiquitination of pRb. The disruption of pRb/E2F regulatory pathways in cells infected with HCV is likely to promote hepatocellular proliferation and chromosomal instability, factors important for the development of liver cancer.

  16. Degradation of peptides and proteins of different sizes by homogenates of human MRC5 lung fibroblasts. Aged cells have a decreased ability to degrade shortened proteins.

    Science.gov (United States)

    Wharton, S A; Hipkiss, A R

    1985-05-20

    The degradation of haemoglobin and haemoglobin-derived peptide fragments by homogenates of MRC5 fibroblasts has been investigated. Results show that the smaller fragments were degraded more rapidly than larger substrates at both pH 5.5 and pH 7.5. Only the smallest of the soluble cyanogen bromide peptides (Mr 3500) was degraded at pH 7.5. Degradation at pH 5.5 proceeded more rapidly than that at pH 7.5 for all substrates tested but was more marked with the larger substrates. Homogenates prepared from aged cells degraded puromycin peptides and, to a lesser extent, cyanogen bromide peptides at a slower rate, at pH 7.5, than those prepared from younger cells. We suggest that cytosolic degradation is less selective and at least one cytosolic proteolytic activity decreases as cells age.

  17. LINGO-1 promotes lysosomal degradation of amyloid-β protein precursor

    Directory of Open Access Journals (Sweden)

    Rian de Laat

    2015-03-01

    Full Text Available Sequential proteolytic cleavages of amyloid-β protein precursor (AβPP by β-secretase and γ-secretase generate amyloid β (Aβ peptides, which are thought to contribute to Alzheimer's disease (AD. Much of this processing occurs in endosomes following endocytosis of AβPP from the plasma membrane. However, this pathogenic mode of processing AβPP may occur in competition with lysosomal degradation of AβPP, a common fate of membrane proteins trafficking through the endosomal system. Following up on published reports that LINGO-1 binds and promotes the amyloidogenic processing of AβPP we have examined the consequences of LINGO-1/AβPP interactions. We report that LINGO-1 and its paralogs, LINGO-2 and LINGO-3, decrease processing of AβPP in the amyloidogenic pathway by promoting lysosomal degradation of AβPP. We also report that LINGO-1 levels are reduced in AD brain, representing a possible pathogenic mechanism stimulating the generation of Aβ peptides in AD.

  18. LINGO-1 promotes lysosomal degradation of amyloid-β protein precursor.

    Science.gov (United States)

    de Laat, Rian; Meabon, James S; Wiley, Jesse C; Hudson, Mark P; Montine, Thomas J; Bothwell, Mark

    2015-01-01

    Sequential proteolytic cleavages of amyloid-β protein precursor (AβPP) by β-secretase and γ-secretase generate amyloid β (Aβ) peptides, which are thought to contribute to Alzheimer's disease (AD). Much of this processing occurs in endosomes following endocytosis of AβPP from the plasma membrane. However, this pathogenic mode of processing AβPP may occur in competition with lysosomal degradation of AβPP, a common fate of membrane proteins trafficking through the endosomal system. Following up on published reports that LINGO-1 binds and promotes the amyloidogenic processing of AβPP we have examined the consequences of LINGO-1/AβPP interactions. We report that LINGO-1 and its paralogs, LINGO-2 and LINGO-3, decrease processing of AβPP in the amyloidogenic pathway by promoting lysosomal degradation of AβPP. We also report that LINGO-1 levels are reduced in AD brain, representing a possible pathogenic mechanism stimulating the generation of Aβ peptides in AD.

  19. Investigating ER-Associated Degradation with RNAi Screening - and Searching for Model Proteins to Do It with

    DEFF Research Database (Denmark)

    Jensen, Njal Winther

    Abstract In eukaryotes, secretory proteins are translocated into the endoplasmic reticulum (ER) for folding assistance, acquisition of posttranslational modifications and sorting. Proteins that do not obtain their native conformation are eliminated by ER-associated degradation (ERAD). ERAD...... is a sophisticated pathway that recognizes misfolded proteins and targets them for degradation by the 26S proteasome residing in the cytosol. More than 60 diseases including Alzheimer’s disease, Huntington’s disease and Parkinson’s disease have been linked to the ERAD pathway underscoring its crucial role...... of this project was to identify endogenous proteins suitable as model proteins. Unfortunately, the methodological approach proved unsuccessful. Instead, mutants of two polytopic membrane proteins were investigated for their potential as ERAD substrates and a mutant of one of these proteins, ATP13A2, turned out...

  20. Ação da própolis sobre a desaminação de aminoácidos e a fermentação ruminal Effect of the propolis on amino acids deamination and ruminal fermentation

    Directory of Open Access Journals (Sweden)

    Deolindo Stradiotti Júnior

    2004-08-01

    Holstein steers were used, in two experimental periods, fed a 35.0% concentrate diet and submitted to the control and propolis extract treatments. The propolis extract obtained with ethanol at 70% in water was in vitro more efficient than that obtained with ethanol at 99.5% in water. Values up to 78% of SAAP inhibition in relation to the control were obtained. The propolis extract did not affect dry matter intake, ruminal pH, ammonia and microbial protein concentrations. There were no differences for molar proportions of the volatile fat acid (VFA, acetic, propionic and butyric in the rumen fluid. However, the propolis extract increased the total VFA concentration as well as inhibited SAAP by the ruminal microorganisms, demonstrating that, in spite of not having reduced the ruminal ammonia level, it seems to exist potential of this effect to happen in other situations, as in diets with high rate of degradable protein/fermentable carbohydrate, observed in new grass pastures or grass pastures consociated with legumes.

  1. Activity dependent protein degradation is critical for the formation and stability of fear memory in the amygdala.

    Directory of Open Access Journals (Sweden)

    Timothy J Jarome

    Full Text Available Protein degradation through the ubiquitin-proteasome system [UPS] plays a critical role in some forms of synaptic plasticity. However, its role in memory formation in the amygdala, a site critical for the formation of fear memories, currently remains unknown. Here we provide the first evidence that protein degradation through the UPS is critically engaged at amygdala synapses during memory formation and retrieval. Fear conditioning results in NMDA-dependent increases in degradation-specific polyubiquitination in the amygdala, targeting proteins involved in translational control and synaptic structure and blocking the degradation of these proteins significantly impairs long-term memory. Furthermore, retrieval of fear memory results in a second wave of NMDA-dependent polyubiquitination that targets proteins involved in translational silencing and synaptic structure and is critical for memory updating following recall. These results indicate that UPS-mediated protein degradation is a major regulator of synaptic plasticity necessary for the formation and stability of long-term memories at amygdala synapses.

  2. Interplay between Molecular Chaperones and the Ubiquitin-Proteasome System in Targeting of Misfolded Proteins for Degradation

    DEFF Research Database (Denmark)

    Poulsen, Esben Guldahl

    interacting with purified 26S proteasomes, and the subsequent characterization of two novel proteasome interacting proteins. The third study was aimed at analyzing the chaperone-assisted pathway leading to degradation of misfolded kinetochore proteins in S. pombe. In this study chaperones, E2s, E3s and DUBs...

  3. Linseed: a valuable feedstuff for ruminants

    Directory of Open Access Journals (Sweden)

    Doreau Michel

    2015-11-01

    Full Text Available Linseeds are used in ruminant feeding for a long time, but this feedstuff knows now increasing interest. Linseeds are rich in alpha-linolenic acid, a fatty acid from the omega-3 series. Despite an extensive biohydrogenation of dietary alpha-linolenic acid in the rumen, its concentration in milk and beef meat increases with linseed incorporation in diets; this increase is accompanied by that of other fatty acids produced during biohydrogenation, especially conjugated linoleic acids and trans 18:1 fatty acids. The increase in cow fertility due to omega-3 fatty acids has not been demonstrated. Furthermore, linseed incorporation in ruminant diets is one of the most efficient ways to decrease enteric methane emissions. In addition to a global mitigating effect of all lipid sources on methane, linseeds have a specific effect due to changes in rumen microbial ecosystem. The practical use of linseeds in ruminant feeding at a large scale requires the absence of negative effect at any step of the ruminant production system. An excessive supply of lipids from linseeds can have deleterious effects on digestive efficiency, milk fat and protein content, beef susceptibility to oxidation, milk and beef fatty acid composition, but when linseed incorporation in the diet does not exceed ca. 3% of additional fat, only positive effects are remaining. A challenge is the increase in linseed cropping to meet increased needs for animal feeding.

  4. Evaluation of Plasma Fibrinogen Degradation Products and Total Serum Protein Concentration in Oral Submucous Fibrosis

    Science.gov (United States)

    B.N.V.S., Satish; B., Maharudrappa; K.M., Prashant; Hugar, Deepa; Allad, Umesh; Prabhu, Prasanth S.

    2014-01-01

    Background: Oral submucous fibrosis (OSMF) is a potentially malignant disorder with a multifactorial etiology. Malnutrition is a major problem for the inhabitants of most countries where OSMF is prevalent. Recently, a new direction in the etiopathogenesis was provided by the identification of fibrinogen degradation products (FDP) in the plasma of OSMF patients. Aims and Objectives: To assess the role of FDP in the etiology of OSMF and to correlate with the nutritional status by evaluating the total serum protein level. The study also determines to evaluate the correlation between the levels of plasma FDP with respect to the staging and grading of OSMF. Correlation between the levels of Total Serum Protein (TSP) with respect to the staging and grading of OSMF was also evaluated. Materials and Methods: The study included 30 cases clinically and histopathologically diagnosed as oral submucous fibrosis. The FDP levels were assessed using both qualitative and semi quantitative method as supplied by ‘Tulip Diagnostics (P) Ltd. Total Serum Protein (TSP) estimation was done by Biuret method using Liquixx Protein kit by Erba, Manheim. Results: The study indicates that in qualitative assessment of FDP only 14 subjects showed the presence of FDP levels>200ng/ml. In semiquantitative assessment there is no significant association between varying clinical stages and histopathological grades and FDP levels. Total serum Protein level showed a marginal increase in all subjects. The study revealed a positive correlation between FDP and TSP in all OSMF subjects. Conclusion: A larger sample size which would be a better representation of the population and the use of different methods which have higher sensitivities and specificities to evaluate FDP level and detailed fractional analysis of protein along with immunoglobulin profiling would facilitate in attaining more conclusive results. PMID:24995245

  5. Protein degradation by ubiquitin–proteasome system in formation and labilization of contextual conditioning memory

    Science.gov (United States)

    Sol Fustiñana, María; de la Fuente, Verónica; Federman, Noel; Freudenthal, Ramiro

    2014-01-01

    The ubiquitin–proteasome system (UPS) of protein degradation has been evaluated in different forms of neural plasticity and memory. The role of UPS in such processes is controversial. Several results support the idea that the activation of this system in memory consolidation is necessary to overcome negative constrains for plasticity. In this case, the inhibition of the UPS during consolidation impairs memory. Similar results were reported for memory reconsolidation. However, in other cases, the inhibition of UPS had no effect on memory consolidation and reconsolidation but impedes the amnesic action of protein synthesis inhibition after retrieval. The last finding suggests a specific action of the UPS inhibitor on memory labilization. However, another interpretation is possible in terms of the synthesis/degradation balance of positive and negative elements in neural plasticity, as was found in the case of long-term potentiation. To evaluate these alternative interpretations, other reconsolidation-interfering drugs than translation inhibitors should be tested. Here we analyzed initially the UPS inhibitor effect in contextual conditioning in crabs. We found that UPS inhibition during consolidation impaired long-term memory. In contrast, UPS inhibition did not affect memory reconsolidation after contextual retrieval but, in fact, impeded memory labilization, blocking the action of drugs that does not affect directly the protein synthesis. To extend these finding to vertebrates, we performed similar experiments in contextual fear memory in mice. We found that the UPS inhibitor in hippocampus affected memory consolidation and blocked memory labilization after retrieval. These findings exclude alternative interpretations to the requirement of UPS in memory labilization and give evidence of this mechanism in both vertebrates and invertebrates. PMID:25135196

  6. Masseter muscle myofibrillar protein synthesis and degradation in an experimental critical illness myopathy model.

    Science.gov (United States)

    Akkad, Hazem; Corpeno, Rebeca; Larsson, Lars

    2014-01-01

    Critical illness myopathy (CIM) is a debilitating common consequence of modern intensive care, characterized by severe muscle wasting, weakness and a decreased myosin/actin (M/A) ratio. Limb/trunk muscles are primarily affected by this myopathy while cranial nerve innervated muscles are spared or less affected, but the mechanisms underlying these muscle-specific differences remain unknown. In this time-resolved study, the cranial nerve innervated masseter muscle was studied in a unique experimental rat intensive care unit (ICU) model, where animals were exposed to sedation, neuromuscular blockade (NMB), mechanical ventilation, and immobilization for durations varying between 6 h and 14d. Gel electrophoresis, immunoblotting, RT-PCR and morphological staining techniques were used to analyze M/A ratios, myofiber size, synthesis and degradation of myofibrillar proteins, and levels of heat shock proteins (HSPs). Results obtained in the masseter muscle were compared with previous observations in experimental and clinical studies of limb muscles. Significant muscle-specific differences were observed, i.e., in the masseter, the decline in M/A ratio and muscle fiber size was small and delayed. Furthermore, transcriptional regulation of myosin and actin synthesis was maintained, and Akt phosphorylation was only briefly reduced. In studied degradation pathways, only mRNA, but not protein levels of MuRF1, atrogin-1 and the autophagy marker LC3b were activated by the ICU condition. The matrix metalloproteinase MMP-2 was inhibited and protective HSPs were up-regulated early. These results confirm that the cranial nerve innervated masticatory muscles is less affected by the ICU-stress response than limb muscles, in accordance with clinical observation in ICU patients with CIM, supporting the model' credibility as a valid CIM model.

  7. Preparation of cell aggregates incorporating gelatin hydrogel microspheres containing bone morphogenic protein-2 with different degradabilities.

    Science.gov (United States)

    Tajima, Shuhei; Tabata, Yasuhiko

    2017-07-31

    The objective of this study is to evaluate the survival and functions of cells in cell aggregates incorporating gelatin hydrogel microspheres (GM) containing bone morphogenic protein-2 (BMP2). Gelatin was dehydrothermally crosslinked in a water-in-oil emulsion state at 140 °C for various time periods to prepare GM with different degradabilities. BMP2 was dropped onto the GM freeze dried, followed by leaving at 25 °C to obtain GM containing BMP2 (GM-BMP2). MC3T3-E1 cells were cultured with GM-BMP2 and GM in round U-bottom wells of 96-multiwell microplates which had been coated with poly (vinyl alcohol) (PVA), to allow to form cell aggregates containing GM-BMP2 and GM, respectively. Higher MC3T3-E1 cell proliferation and the L-lactic acid/glucose ratio were observed for MC3T3-E1 cell aggregates cultured with the GM of slower degradation. The runt-related transcription factor 2 (RUNX2) messenger ribonucleic acid (mRNA) expression, alkaline phosphatase (ALP) activity, and calcium content of MC3T3-E1 cells in cell aggregates were assayed to evaluate their osteogenic differentiation. When cultured for 7 days with GM-BMP2 or free BMP2, the RUNX2 mRNA expression and ALP activity were higher for MC3T3-E1 cell aggregates cultured with the GM-BMP2 of faster degradation than those of free BMP2 added into the medium. After 21 days culture, the ALP activity and calcium content were higher for the GM-BMP2 of medium degradation compared with other experimental groups. It is concluded that BMP2 of GM-BMP2 incorporated in the cell aggregates enhanced the osteogenic differentiation of cells compared with free BMP2 added externally. The degradability of GM-BMP2 affected the extent of osteogenic differentiation.

  8. Pu-Erh Tea Extract Induces the Degradation of FET Family Proteins Involved in the Pathogenesis of Amyotrophic Lateral Sclerosis

    Directory of Open Access Journals (Sweden)

    Yang Yu

    2014-01-01

    Full Text Available FET family proteins consist of fused in sarcoma/translocated in liposarcoma (FUS/TLS, Ewing's sarcoma (EWS, and TATA-binding protein-associated factor 15 (TAF15. Mutations in the copper/zinc superoxide dismutase (SOD1, TAR DNA-binding protein 43 (TDP-43, and FET family proteins are associated with the development of amyotrophic lateral sclerosis (ALS, a fatal neurodegenerative disease. There is currently no cure for this disease and few effective treatments are available. Epidemiological studies indicate that the consumption of tea is associated with a reduced risk of developing neurodegenerative diseases. The results of this study revealed that components of a pu-erh tea extract (PTE interacted with FET family proteins but not with TDP-43 or SOD1. PTE induced the degradation of FET family proteins but had no effects on TDP-43 or SOD1. The most frequently occurring ALS-linked FUS/TLS mutant protein, R521C FUS/TLS, was also degraded in the presence of PTE. Furthermore, ammonium chloride, a lysosome inhibitor, but not lactacystin, a proteasome inhibitor, reduced the degradation of FUS/TLS protein by PTE. PTE significantly reduced the incorporation of R521C FUS/TLS into stress granules under stress conditions. These findings suggest that PTE may have beneficial health effects, including preventing the onset of FET family protein-associated neurodegenerative diseases and delaying the progression of ALS by inhibiting the cytoplasmic aggregation of FET family proteins.

  9. Pu-Erh Tea Extract Induces the Degradation of FET Family Proteins Involved in the Pathogenesis of Amyotrophic Lateral Sclerosis

    Science.gov (United States)

    Cai, Xianbin; Fang, Chongye; Shi, Wei; Sheng, Jun

    2014-01-01

    FET family proteins consist of fused in sarcoma/translocated in liposarcoma (FUS/TLS), Ewing's sarcoma (EWS), and TATA-binding protein-associated factor 15 (TAF15). Mutations in the copper/zinc superoxide dismutase (SOD1), TAR DNA-binding protein 43 (TDP-43), and FET family proteins are associated with the development of amyotrophic lateral sclerosis (ALS), a fatal neurodegenerative disease. There is currently no cure for this disease and few effective treatments are available. Epidemiological studies indicate that the consumption of tea is associated with a reduced risk of developing neurodegenerative diseases. The results of this study revealed that components of a pu-erh tea extract (PTE) interacted with FET family proteins but not with TDP-43 or SOD1. PTE induced the degradation of FET family proteins but had no effects on TDP-43 or SOD1. The most frequently occurring ALS-linked FUS/TLS mutant protein, R521C FUS/TLS, was also degraded in the presence of PTE. Furthermore, ammonium chloride, a lysosome inhibitor, but not lactacystin, a proteasome inhibitor, reduced the degradation of FUS/TLS protein by PTE. PTE significantly reduced the incorporation of R521C FUS/TLS into stress granules under stress conditions. These findings suggest that PTE may have beneficial health effects, including preventing the onset of FET family protein-associated neurodegenerative diseases and delaying the progression of ALS by inhibiting the cytoplasmic aggregation of FET family proteins. PMID:24804206

  10. Rabbit Feces as Feed for Ruminants and as an Energy Source

    Directory of Open Access Journals (Sweden)

    Pier Giorgio Peiretti

    2014-12-01

    Full Text Available There are prospects for using novel feeds from various sources to provide ruminants with alternative sources of protein and energy such as by-products, and animal wastes. Rabbit feces are a concentrated source of fiber and could have commercial potential both as input biomass in anaerobic processes for biogas production, as well as a fibrous source for ruminal degradation. The aims of this work were to assess the potential as ruminant feeding and as biogas production of rabbit feces, in comparison with 12 crops. The chemical composition and the potential and experimental in vitro true digestibility (IVTD and neutral detergent fiber digestibility (NDFD of 148 feces samples were determined by using chemical methods, Daisy system digestibility and/or NIRS predictions. The average biomethane potential (BMP was 286 ± 10 lCH4/kg SV with −4% vs. the crops average. Milk forage unit (milk FU, IVTD and NDFD of feces were 0.54 ± 0.06 milk FU/kg DM, 74% ± 3% and 50% ± 5%, respectively, with comparisons of −19%, −11% and −24% vs. the crops average. Reconstruction of the potential values based on the chemical constituents but using the crop partial least square model well agreed with the NIRS calibrations and cross-validation. In a global NIRS calibration of the feces and crops the relative predicted deviation for IVTD, NDFD and milk FU were 3.1, 2.9 and 2.6, respectively, and only 1.5 for BMP. Running the Daisy system for rabbit feces in rumen fluid gave some inconsistencies, weakened the functional relationships, and appeared not to be correlated with the potential values of IVTD and NDFD. Nevertheless, the energetic potential of feces appears to be similar to some conventional crops at different degrees of maturity. Thus we conclude that rabbit feces has potential value as a ruminant feed and for biogas production.

  11. Microwave irradiation induced changes in protein molecular structures of barley grains: relationship to changes in protein chemical profile, protein subfractions, and digestion in dairy cows.

    Science.gov (United States)

    Yan, Xiaogang; Khan, Nazir A; Zhang, Fangyu; Yang, Ling; Yu, Peiqiang

    2014-07-16

    The objectives of this study were to evaluate microwave irradiation (MIR) induced changes in crude protein (CP) subfraction profiles, ruminal CP degradation characteristics and intestinal digestibility of rumen undegraded protein (RUP), and protein molecular structures in barley (Hordeum vulgare) grains. Samples from hulled (n = 1) and hulless cultivars (n = 2) of barley, harvested from four replicate plots in two consecutive years, were evaluated. The samples were either kept as raw or irradiated in a microwave for 3 min (MIR3) or 5 min (MIR5). Compared to raw grains, MIR5 decreased the contents of rapidly degradable CP subfraction (from 45.22 to 6.36% CP) and the ruminal degradation rate (from 8.16 to 3.53%/h) of potentially degradable subfraction. As a consequence, the effective ruminal degradability of CP decreased (from 55.70 to 34.08% CP) and RUP supply (from 43.31 to 65.92% CP) to the postruminal tract increased. The MIR decreased the spectral intensities of amide 1, amide II, α-helix, and β-sheet and increased their ratios. The changes in protein spectral intensities were strongly correlated with the changes in CP subfractions and digestive kinetics. These results show that MIR for a short period (5 min) with a lower energy input can improve the nutritive value and utilization of CP in barely grains.

  12. Ruminal acidosis: A review with detailed reference to the controlling ...

    African Journals Online (AJOL)

    Apart from stepwise transition from roughage to concentrates, methods adopted to reduce SARA include grain treatment to reduce starch degradation, feed additives such as buffers to control ruminal pH, dicarboxylic acids to stimulate the growth of lactate utilisers, antibiotics such as virginiamycin and the ionophores which ...

  13. A comparison of lysosomal involvements in the degradation of normal and abnormal endogenous proteins of differing half-lives in MRC5 cells.

    Science.gov (United States)

    Wharton, S A; Riley, P A

    1983-05-15

    Protein degradation by diploid human-embryo lung fibroblasts (MRC5 cells) in monolayer culture was studied. 1. Varying the labelling period of proteins was found to alter the half-lives of labelled abnormal canavanine-containing proteins to an extent very similar to that obtained with normal proteins. 2. By manipulating the times of labelling it was possible to generate a species of abnormal protein with a greater half-life than that of a species of normal protein. A comparison of the lysosomal involvement in their degradation as determined both by inhibition by methylamine, a lysosomotropic agent, and by the degree of increase in protein degradation in step-down conditions, indicated that the degree of lysosomal involvement was not entirely dependent upon the half-life of the protein, but that abnormal proteins are preferentially degraded non-lysosomally. 3. The microtubule inhibitors colchicine and vinblastine were found to stimulate statistically basal protein degradation of normal long-labelled protein, whereas they had less effect upon the basal degradation of the other species of proteins studied and very little effect upon step-down degradation of all proteins studied. The stimulation in protein degradation found did not seem to involve the acid proteinases of lysosomes.

  14. 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 ...

  15. Pseudomonas aeruginosa elastase provides an escape from phagocytosis by degrading the pulmonary surfactant protein-A.

    Directory of Open Access Journals (Sweden)

    Zhizhou Kuang

    Full Text Available Pseudomonas aeruginosa is an opportunistic pathogen that causes both acute pneumonitis in immunocompromised patients and chronic lung infections in individuals with cystic fibrosis and other bronchiectasis. Over 75% of clinical isolates of P. aeruginosa secrete elastase B (LasB, an elastolytic metalloproteinase that is encoded by the lasB gene. Previously, in vitro studies have demonstrated that LasB degrades a number of components in both the innate and adaptive immune systems. These include surfactant proteins, antibacterial peptides, cytokines, chemokines and immunoglobulins. However, the contribution of LasB to lung infection by P. aeruginosa and to inactivation of pulmonary innate immunity in vivo needs more clarification. In this study, we examined the mechanisms underlying enhanced clearance of the ΔlasB mutant in mouse lungs. The ΔlasB mutant was attenuated in virulence when compared to the wild-type strain PAO1 during lung infection in SP-A+/+ mice. However, the ΔlasB mutant was as virulent as PAO1 in the lungs of SP-A⁻/⁻ mice. Detailed analysis showed that the ΔlasB mutant was more susceptible to SP-A-mediated opsonization but not membrane permeabilization. In vitro and in vivo phagocytosis experiments revealed that SP-A augmented the phagocytosis of ΔlasB mutant bacteria more efficiently than the isogenic wild-type PAO1. The ΔlasB mutant was found to have a severely reduced ability to degrade SP-A, consequently making it unable to evade opsonization by the collectin during phagocytosis. These results suggest that P. aeruginosa LasB protects against SP-A-mediated opsonization by degrading the collectin.

  16. Role of phosphorolytic cleavage in cellobiose and cellodextrin metabolism by the ruminal bacterium Prevotella ruminicola.

    Science.gov (United States)

    Lou, J; Dawson, K A; Strobel, H J

    1996-05-01

    In bacteria, cellobiose and cellodextrins are usually degraded by either hydrolytic or phosphorolytic cleavage. Prevotella ruminicola B(1)4 is a noncellulolytic ruminal bacterium which has the ability to utilize the products of cellulose degradation. In this organism, cellobiose hydrolytic cleavage activity was threefold greater than phosphorolytic cleavage activity (113 versus 34 nmol/min/mg of protein), as measured by an enzymatic assay. Cellobiose phosphorylase activity (measured as the release of P(i)) was found in cellobiose-, mannose-, xylose-, lactose-, and cellodextrin-grown cells (> 92 nmol of P(i)/min/mg of protein), but the activity was reduced by more than 74% for cells grown on fructose, L-arabinose, sucrose, maltose, or glucose. A small amount of cellodextrin phosphorylase activity (19 nmol/min/mg of protein) was also detected, and both phosphorylase activities were located in the cytoplasm. Degradation involving phosphorolytic cleavage conserves more metabolic energy than simple hydrolysis, and such degradation is consistent with substrate-limiting conditions such as those often found in the rumen.

  17. Uptake of proteins and degradation of human serum albumin by Plasmodium falciparum – infected human erythrocytes

    Directory of Open Access Journals (Sweden)

    Malhotra Pawan

    2003-05-01

    Full Text Available Abstract Background Intraerythrocytic malaria parasites actively import obligate nutrients from serum and export proteins and lipids to erythrocyte cytoplasm and membrane. The import of macromolecules in the malaria parasite has been the subject of many debates. To understand the import of macromolecules by the parasite, we studied the uptake of proteins by Plasmodium falciparum infected human erythrocyte. Methods Proteins were biotin labelled individually, purified on a gel filtration column and added to uninfected and infected asynchronized culture. The uptake of these proteins by malaria parasites was determined by western blot analysis of parasite pellet and their different fractions using streptavidin-horseradish conjugate. To further confirm this import, we studied the uptake of 125I-labelled proteins by western blot analysis as well as used direct immunofluorescence method. Results Here we show that biotin labelled and radio-iodinated polypeptides of molecular sizes in the range of 45 to 206 kDa, when added in the culture medium, get direct access to the parasite membrane through a membrane network by by-passing the erythrocyte cytosol. The import of these polypeptides is ATP-dependent as sodium azide treatment blocks this uptake. We also show that malaria parasites have the ability to take up and degrade biotin labelled human serum albumin, which has been shown to be essential for the parasite growth. Conclusions These results can be used, as a basis to explore the role of human serum albumin in the intraerythrocytic development of parasites, and this in turn can be an important adjunct to the development of novel antimalarial drugs.

  18. 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

  19. Differential Expression in Phanerochaete chrysosporium of Membrane-Associated Proteins Relevant to Lignin Degradation

    Energy Technology Data Exchange (ETDEWEB)

    Shary, Semarjit; Kapich, Alexander N.; Panisko, Ellen A.; Magnuson, Jon K.; Cullen, Dan; Hammel, Ken

    2008-10-02

    Fungal lignin-degrading systems must include membrane-associated proteins that participate in diverse processes such as uptake and oxidation of lignin fragments, secretion of ligninolytic secondary metabolites, and defense of the mycelium against ligninolytic oxidants. Despite their importance, little is known about the nature or regulation of these membrane-associated components. We grew the white rot basidiomycete Phanerochaete chrysosporium on cellulose or glucose as the carbon source and monitored the mineralization of a 14C-labeled synthetic lignin by these cultures to assess their ligninolytic competence. The results showed that the cellulose-grown cultures were ligninolytic, whereas the glucose-grown ones were not. We isolated microsomal membrane fractions from both types of culture and analyzed tryptic digests of them by shotgun liquid chromatography/tandem mass spectrometry. Comparison of the results against the predicted P. chrysosporium proteome showed that a catalase (Joint Genome Institute P. chrysosporium protein I.D. 124398), an alcohol oxidase (126879), two transporters (137220 and 132234), and two cytochrome P450s (5011 and 8912) were up-regulated under ligninolytic conditions. Real time reverse transcription polymerase chain reaction assays showed that RNA transcripts encoding all of these proteins were also up-regulated in ligninolytic cultures. Catalase 124398, alcohol oxidase 126879, and transporter 137220 were found in a proteomic analysis of partially purified plasma membranes from ligninolytic P. chrysosporium, and are therefore most likely associated with the outer envelope of the fungus.

  20. Identification and Characterization of a Large Protein Essential for Degradation of the Crystalline Region of Cellulose by Cytophaga hutchinsonii.

    Science.gov (United States)

    Wang, Sen; Zhao, Dong; Bai, Xinfeng; Zhang, Weican; Lu, Xuemei

    2017-01-01

    Cytophaga hutchinsonii is a Gram-negative bacterium that can efficiently degrade crystalline cellulose by a unique mechanism different from the free cellulase or cellulosome strategy. In this study, chu_3220, encoding the hypothetical protein CHU_3220 (205 kDa), was identified by insertional mutation and gene deletion as the first gene essential for degradation of the crystalline region but not the amorphous region of cellulose by C. hutchinsonii A chu_3220 deletion mutant was defective in the degradation of crystalline cellulose and increased the degree of crystallinity of Avicel PH101 but could still degrade amorphous cellulose completely. CHU_3220 was found to be located on the outer surface of the outer membrane and could bind to cellulose. It contains 15 PbH1 domains and a C-terminal domain (CHU_C) that was proved to be critical for the localization of CHU_3220 on the cell surface and the function of CHU_3220 in crystalline cellulose degradation. Moreover, the degradation of crystalline cellulose was intact-cell dependent and inhibited by NaN3 Further study showed that chu_3220 was induced by cellulose and that the endoglucanase activity on the cell surface was significantly reduced without chu_3220 Real-time PCR revealed that the transcription of most genes encoding endoglucanases located on the cell surface was decreased in the chu_3220 deletion mutant, indicating that chu_3220 might also play a role in the regulation of the expression of some endoglucanases. Cytophaga hutchinsonii could efficiently degrade crystalline cellulose with a unique mechanism without cellulosomes and free cellulases. It lacks proteins that are thought to play important roles in disruption of the crystalline region of cellulose, including exoglucanases, lytic polysaccharide monooxygenases, expansins, expansin-like proteins, or swollenins, and most of its endoglucanases lack carbohydrate binding modules. The mechanism of the degradation of crystalline cellulose is still unknown. In

  1. Aspergillus protein degradation pathways with different secreted protease sets at neutral and acidic pH.

    Science.gov (United States)

    Sriranganadane, Dev; Waridel, Patrice; Salamin, Karine; Reichard, Utz; Grouzmann, Eric; Neuhaus, Jean-Marc; Quadroni, Manfredo; Monod, Michel

    2010-07-02

    Aspergillus fumigatus grows well at neutral and acidic pH in a medium containing protein as the sole nitrogen source by secreting two different sets of proteases. Neutral pH favors the secretion of neutral and alkaline endoproteases, leucine aminopeptidases (Laps) which are nonspecific monoaminopeptidases, and an X-prolyl dipeptidase (DppIV). Acidic pH environment promotes the secretion of an aspartic endoprotease of pepsin family (Pep1) and tripeptidyl-peptidases of the sedolisin family (SedB and SedD). A novel prolyl peptidase, AfuS28, was found to be secreted in both alkaline and acidic conditions. In previous studies, Laps were shown to degrade peptides from their N-terminus until an X-Pro sequence acts as a stop signal. X-Pro sequences can be then removed by DppIV, which allows Laps access to the following residues. We have shown that at acidic pH Seds degrade large peptides from their N-terminus into tripeptides until Pro in P1 or P'1 position acts as a stop for these exopeptidases. However, X-X-Pro and X-X-X-Pro sequences can be removed by AfuS28 thus allowing Seds further sequential proteolysis. In conclusion, both alkaline and acidic sets of proteases contain exoprotease activity capable of cleaving after proline residues that cannot be removed during sequential digestion by nonspecific exopeptidases.

  2. Chemical biology based on target-selective degradation of proteins and carbohydrates using light-activatable organic molecules.

    Science.gov (United States)

    Toshima, Kazunobu

    2013-05-01

    Proteins and carbohydrates play crucial roles in a wide range of biological processes, including serious diseases. The development of novel and innovative methods for selective control of specific proteins and carbohydrates functions has attracted much attention in the field of chemical biology. In this account article, the development of novel chemical tools, which can degrade target proteins and carbohydrates by irradiation with a specific wavelength of light under mild conditions without any additives, is introduced. This novel class of photochemical agents promise bright prospects for finding not only molecular-targeted bioprobes for understanding of the structure-activity relationships of proteins and carbohydrates but also novel therapeutic drugs targeting proteins and carbohydrates.

  3. Effect of ethephon on protein degradation and the accumulation of pathogensis-related (PR) proteins in tomato leaf discs. [Lycopersicon esculentum

    Energy Technology Data Exchange (ETDEWEB)

    Vera, P.; Conejero, V. (Universidad Politecnica de Valencia (Spain))

    1990-01-01

    The effect of ethephon (2-chloroetylphosphonic acid) on the degradation of proteins and on the induction of Lycopersicon esculentum pathogenesis-related (PR) proteins was studied in tomato leaf discs. The rate of ribulose, -1,5-bisphosphate carboxylase/oxygenase (Rubisco) degradation was maximal in discs after 48 hours of incubation with 1 millimolar ethephon, leading to complete disappearance of Rubisco after 96 hours. This effect was correlated with an increase in PR protein synthesis and the induction of the previously reported alkaline proteolytic enzyme PR-P69. In vivo pulse-chase experiments demonstrated that ethephon not only affected Rubisco content but that of many other {sup 35}S-labeled proteins as well, indicating that ethylene activates a general and nonspecific mechanism of protein degradation. This effect was partially inhibited in vivo by the action of pCMB, a selective inhibitor of cysteine-proteinases such as P69. These data reinforce the hypothesis that P69 and perhaps other PR proteins are involved in the mechanism of accelerated protein degradation activated by ethylene.

  4. Stay-green regulates chlorophyll and chlorophyll-binding protein degradation during senescence.

    Science.gov (United States)

    Hörtensteiner, Stefan

    2009-03-01

    Stay-green mutants are delayed in leaf senescence and have been identified from different plant species, including many crops. Functional stay-greens have the potential to increase plant productivity. In cosmetic stay-greens, however, retention of chlorophyll during senescence is uncoupled from a decline of photosynthetic capacity in these mutants. For many cosmetic stay-green mutants, including Gregor Mendel's famous green cotyledon pea variety, molecular defects were recently identified in orthologous stay-green genes. Stay-green genes encode members of a new family of chloroplast-located proteins, which are likely to function in dismantling of photosynthetic chlorophyll-apoprotein complexes. Their activity is considered as a prerequisite for both chlorophyll and apoprotein degradation during senescence.

  5. Study of messenger RNA inactivation and protein degradation in an Escherichia coli cell-free expression system

    Directory of Open Access Journals (Sweden)

    Noireaux Vincent

    2010-07-01

    Full Text Available Abstract Background A large amount of recombinant proteins can be synthesized in a few hours with Escherichia coli cell-free expression systems based on bacteriophage transcription. These cytoplasmic extracts are used in many applications that require large-scale protein production such as proteomics and high throughput techniques. In recent years, cell-free systems have also been used to engineer complex informational processes. These works, however, have been limited by the current available cell-free systems, which are not well adapted to these types of studies. In particular, no method has been proposed to increase the mRNA inactivation rate and the protein degradation rate in cell-free reactions. The construction of in vitro informational processes with interesting dynamics requires a balance between mRNA and protein synthesis (the source, and mRNA inactivation and protein degradation (the sink. Results Two quantitative studies are presented to characterize and to increase the global mRNA inactivation rate, and to accelerate the degradation of the synthesized proteins in an E. coli cell-free expression system driven by the endogenous RNA polymerase and sigma factor 70. The E. coli mRNA interferase MazF was used to increase and to adjust the mRNA inactivation rate of the Firefly luciferase (Luc and of the enhanced green fluorescent protein (eGFP. Peptide tags specific to the endogenous E. coli AAA + proteases were used to induce and to adjust the protein degradation rate of eGFP. Messenger RNA inactivation rate, protein degradation rate, maturation time of Luc and eGFP were measured. Conclusions The global mRNA turnover and the protein degradation rate can be accelerated and tuned in a biologically relevant range in a cell-free reaction with quantitative procedures easy to implement. These features broaden the capabilities of cell-free systems with a better control of gene expression. This cell-free extract could find some applications in

  6. Parkin Promotes Degradation of the Mitochondrial Pro-Apoptotic ARTS Protein

    Science.gov (United States)

    Kemeny, Stav; Dery, Dikla; Loboda, Yelena; Rovner, Marshall; Lev, Tali; Zuri, Dotan; Finberg, John P. M.; Larisch, Sarit

    2012-01-01

    Parkinson’s disease (PD) is associated with excessive cell death causing selective loss of dopaminergic neurons. Dysfunction of the Ubiquitin Proteasome System (UPS) is associated with the pathophysiology of PD. Mutations in Parkin which impair its E3-ligase activity play a major role in the pathogenesis of inherited PD. ARTS (Sept4_i2) is a mitochondrial protein, which initiates caspase activation upstream of cytochrome c release in the mitochondrial apoptotic pathway. Here we show that Parkin serves as an E3-ubiquitin ligase to restrict the levels of ARTS through UPS-mediated degradation. Though Parkin binds equally to ARTS and Sept4_i1 (H5/PNUTL2), the non-apoptotic splice variant of Sept4, Parkin ubiquitinates and degrades only ARTS. Thus, the effect of Parkin on ARTS is specific and probably related to its pro-apoptotic function. High levels of ARTS are sufficient to promote apoptosis in cultured neuronal cells, and rat brains treated with 6-OHDA reveal high levels of ARTS. However, over-expression of Parkin can protect cells from ARTS-induced apoptosis. Furthermore, Parkin loss-of-function experiments reveal that reduction of Parkin causes increased levels of ARTS and apoptosis. We propose that in brain cells in which the E3-ligase activity of Parkin is compromised, ARTS levels increase and facilitate apoptosis. Thus, ARTS is a novel substrate of Parkin. These observations link Parkin directly to a pro-apoptotic protein and reveal a novel connection between Parkin, apoptosis, and PD. PMID:22792159

  7. Ubiquitination and degradation of the hominoid-specific oncoprotein TBC1D3 is regulated by protein palmitoylation

    Energy Technology Data Exchange (ETDEWEB)

    Kong, Chen; Lange, Jeffrey J.; Samovski, Dmitri [Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110 (United States); Su, Xiong [Department of Internal Medicine, Center for Human Nutrition Washington University School of Medicine, St. Louis, MO 63110 (United States); Liu, Jialiu [Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110 (United States); Sundaresan, Sinju [Department of Internal Medicine, Center for Human Nutrition Washington University School of Medicine, St. Louis, MO 63110 (United States); Stahl, Philip D., E-mail: pstahl@wustl.edu [Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110 (United States)

    2013-05-03

    Highlights: •Hominoid-specific oncogene TBC1D3 is targeted to plasma membrane by palmitoylation. •TBC1D3 is palmitoylated on two cysteine residues: 318 and 325. •TBC1D3 palmitoylation governs growth factors-induced TBC1D3 degradation. •Post-translational modifications may regulate oncogenic properties of TBC1D3. -- Abstract: Expression of the hominoid-specific oncoprotein TBC1D3 promotes enhanced cell growth and proliferation by increased activation of signal transduction through several growth factors. Recently we documented the role of CUL7 E3 ligase in growth factors-induced ubiquitination and degradation of TBC1D3. Here we expanded our study to discover additional molecular mechanisms that control TBC1D3 protein turnover. We report that TBC1D3 is palmitoylated on two cysteine residues: 318 and 325. The expression of double palmitoylation mutant TBC1D3:C318/325S resulted in protein mislocalization and enhanced growth factors-induced TBC1D3 degradation. Moreover, ubiquitination of TBC1D3 via CUL7 E3 ligase complex was increased by mutating the palmitoylation sites, suggesting that depalmitoylation of TBC1D3 makes the protein more available for ubiquitination and degradation. The results reported here provide novel insights into the molecular mechanisms that govern TBC1D3 protein degradation. Dysregulation of these mechanisms in vivo could potentially result in aberrant TBC1D3 expression and promote oncogenesis.

  8. PLAA Mutations Cause a Lethal Infantile Epileptic Encephalopathy by Disrupting Ubiquitin-Mediated Endolysosomal Degradation of Synaptic Proteins.

    Science.gov (United States)

    Hall, Emma A; Nahorski, Michael S; Murray, Lyndsay M; Shaheen, Ranad; Perkins, Emma; Dissanayake, Kosala N; Kristaryanto, Yosua; Jones, Ross A; Vogt, Julie; Rivagorda, Manon; Handley, Mark T; Mali, Girish R; Quidwai, Tooba; Soares, Dinesh C; Keighren, Margaret A; McKie, Lisa; Mort, Richard L; Gammoh, Noor; Garcia-Munoz, Amaya; Davey, Tracey; Vermeren, Matthieu; Walsh, Diana; Budd, Peter; Aligianis, Irene A; Faqeih, Eissa; Quigley, Alan J; Jackson, Ian J; Kulathu, Yogesh; Jackson, Mandy; Ribchester, Richard R; von Kriegsheim, Alex; Alkuraya, Fowzan S; Woods, C Geoffrey; Maher, Eamonn R; Mill, Pleasantine

    2017-05-04

    During neurotransmission, synaptic vesicles undergo multiple rounds of exo-endocytosis, involving recycling and/or degradation of synaptic proteins. While ubiquitin signaling at synapses is essential for neural function, it has been assumed that synaptic proteostasis requires the ubiquitin-proteasome system (UPS). We demonstrate here that turnover of synaptic membrane proteins via the endolysosomal pathway is essential for synaptic function. In both human and mouse, hypomorphic mutations in the ubiquitin adaptor protein PLAA cause an infantile-lethal neurodysfunction syndrome with seizures. Resulting from perturbed endolysosomal degradation, Plaa mutant neurons accumulate K63-polyubiquitylated proteins and synaptic membrane proteins, disrupting synaptic vesicle recycling and neurotransmission. Through characterization of this neurological intracellular trafficking disorder, we establish the importance of ubiquitin-mediated endolysosomal trafficking at the synapse. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

  9. Degradation of Amino Acids and Structure in Model Proteins and Bacteriophage MS2 by Chlorine, Bromine, and Ozone.

    Science.gov (United States)

    Choe, Jong Kwon; Richards, David H; Wilson, Corey J; Mitch, William A

    2015-11-17

    Proteins are important targets of chemical disinfectants. To improve the understanding of disinfectant-protein reactions, this study characterized the disinfectant:protein molar ratios at which 50% degradation of oxidizable amino acids (i.e., Met, Tyr, Trp, His, Lys) and structure were observed during HOCl, HOBr, and O3 treatment of three well-characterized model proteins and bacteriophage MS2. A critical question is the extent to which the targeting of amino acids is driven by their disinfectant rate constants rather than their geometrical arrangement. Across the model proteins and bacteriophage MS2 (coat protein), differing widely in structure, methionine was preferentially targeted, forming predominantly methionine sulfoxide. This targeting concurs with its high disinfectant rate constants and supports its hypothesized role as a sacrificial antioxidant. Despite higher HOCl and HOBr rate constants with histidine and lysine than for tyrosine, tyrosine generally was degraded in preference to histidine, and to a lesser extent, lysine. These results concur with the prevalence of geometrical motifs featuring histidines or lysines near tyrosines, facilitating histidine and lysine regeneration upon Cl[+1] transfer from their chloramines to tyrosines. Lysine nitrile formation occurred at or above oxidant doses where 3,5-dihalotyrosine products began to degrade. For O3, which lacks a similar oxidant transfer pathway, histidine, tyrosine, and lysine degradation followed their relative O3 rate constants. Except for its low reactivity with lysine, the O3 doses required to degrade amino acids were as low as or lower than for HOCl or HOBr, indicating its oxidative efficiency. Loss of structure did not correlate with loss of particular amino acids, suggesting the need to characterize the oxidation of specific geometric motifs to understand structural degradation.

  10. Comparison of protein degradation, protein oxidation, and μ-calpain activation between pale, soft, and exudative and red, firm, and nonexudative pork during postmortem aging.

    Science.gov (United States)

    Yin, Y; Zhang, W G; Zhou, G H; Guo, B

    2014-08-01

    The objective of this study was to investigate the differences in protein modifications between pale, soft, and exudative (PSE) and red, firm, and nonexudative (RFN) pork during postmortem (PM) aging. Longissimus dorsi (LD) including 8 PSE and 8 RFN muscles were individually removed from 16 carcasses. These 16 LD muscles were vacuum packaged at 24 h after slaughter and stored at 4°C for 1, 3, and 5 d. The centrifugation loss, drip loss, color, protein solubility, protein oxidation, protein degradation including desmin, troponin T, and integrin, and μ-calpain activation were determined. The pH of PSE samples was significantly lower than that of RFN samples at both 1 and 24 h PM (P 0.05). In addition, PSE pork presented a lower solubility of sarcoplasmic protein, myofibrillar protein, and total protein than RFN pork except the solubility of myofibrillar protein at d 1 (P degradation products of integrin were detected in PSE pork compared to that of RFN pork at d 1 (P Red, firm, and nonexudative pork presented lower intensity of intact 80 kDa calpain and greater intensity of autolyzed 76 kDa product compared to PSE pork (P degradation including desmin and integrin, and the level of protein solubility in PSE pork could contribute to its low water holding capacity during PM storage.

  11. The implications of condensed tannins on the nutritive value of temperate forages fed to ruminants.

    Science.gov (United States)

    Barry, T N; McNabb, W C

    1999-04-01

    New methodology for measuring forage condensed tannin (CT) content is described and the effects of CT upon forage feeding and nutritive value for ruminant animals are reviewed. CT react with forage proteins in a pH-reversible manner, with reactivity determined by the concentration, structure and molecular mass of the CT. Increasing concentrations of CT in Lotus corniculatus and Lotus pedunculatus reduce the rates of solubilization and degradation of fraction 1 leaf protein in the rumen and increase duodenal non-NH3 N flow. Action of medium concentrations of total CT in Lotus corniculatus (30-40 g/kg DM) increased the absorption of essential amino acids from the small intestine and increased wool growth, milk secretion and reproductive rate in grazing sheep without affecting voluntary feed intake, thus improving the efficiency of food conversion. High concentrations of CT in Lotus pedunculatus (75-100 g/kg DM) depressed voluntary feed intake and rumen carbohydrate digestion and depressed rates of body and wool growth in grazing sheep. The minimum concentration of CT to prevent rumen frothy bloat in cattle is defined as 5 g/kg DM and sheep grazing CT-containing legumes were shown to better tolerate internal parasite infections than sheep grazing non CT-containing forages. It was concluded that defined concentrations of forage CT can be used to increase the efficiencies of protein digestion and animal productivity in forage-fed ruminants and to develop more ecologically sustainable systems of controlling some diseases under grazing.

  12. Microbial degradation of tannins--a current perspective.

    Science.gov (United States)

    Bhat, T K; Singh, B; Sharma, O P

    1998-01-01

    Tannins are water-soluble polyphenolic compounds having wide prevalence in plants. Hydrolysable and condensed tannins are the two major classes of tannins. These compounds have a range of effects on various organisms--from toxic effects on animals to growth inhibition of microorganisms. Some microbes are, however, resistant to tannins, and have developed various mechanisms and pathways for tannin degradation in their natural milieu. The microbial degradation of condensed tannins is, however, less than hydrolysable tannins in both aerobic and anaerobic environments. A number of microbes have also been isolated from the gastrointestinal tract of animals, which have the ability to break tannin-protein complexes and degrade tannins, especially hydrolysable tannins. Tannase, a key enzyme in the degradation of hydrolysable tannins, is present in a diverse group of microorganisms, including rumen bacteria. This enzyme is being increasingly used in a number of processes. Presently, there is a need for increased understanding of the biodegradation of condensed tannins, particularly in ruminants.

  13. Composição química e cinética da degradação ruminal de gramíneas do gênero Cynodon em diferentes idades ao corte - DOI: 10.4025/actascianimsci.v27i2.1221 Chemical composition and ruminal degradation kinetic of Cynodon grasses in different cut age- DOI: 10.4025/actascianimsci.v27i2.1221

    Directory of Open Access Journals (Sweden)

    Ulysses Cecato

    2005-03-01

    Full Text Available O experimento teve por objetivos avaliar a composição química, estimar a energia metabolizável, a digestibilidade in vitro da matéria seca (DIVMS e da matéria orgânica (DIVMO com o uso do rúmen artificial desenvolvido pela Ankom e, também determinar a degradabilidade in situ da MS (DEMS de três cultivares do gênero Cynodon (Tifton 44, Tifton 85 e Coast-cross, colhidos com idades de 21, 42 e 63 dias no verão. Para a degradabilidade in situ utilizou-se três vacas com fístulas ruminais, com tempos de incubação de 96, 48, 12, 6 e 0 hora. No entanto, o tempo 0 foi determinado através da lavagem dos sacos com água em máquina de lavar roupas. Utilizou-se o delineamento experimental de parcelas subdivididas (gramíneas como parcelas e idade ao corte como sub-parcelas com três repetições. Para o fator idade ao corte, foi usado regressão e os modelos foram escolhidos baseados na análise de identidade. Houve aumento (p 0,05 para a taxa de degradação (c, da MS. A degradabilidade efetiva da MS com taxas de passagem de 2, 5 e 8%/h foi influenciada (p The objectives of the experiment were to estimate the chemical composition, to estimate the metabolically energy, dry matter in vitro digestibility (DMIVD, organic matter in vitro digestibility (OMIVD through an artificial rumen developed by Ankom and also dry matter in situ degradability (DMISD of three Cynodon grasses (Tifton 85, Tifton 44 and Coast-cross collected at the ages of 21, 42 and 63 days in the summer. Three cows with ruminal fistula were used to determine the in situ degradability. The following incubation times were 96, 48, 12, 6, 0 hours. However, the zero time was determinate through of washing with water in washing machine. The experimental design was a split-plot (plots like grasses and sub-plots like age of cut with three repetitions. The cut age factor was evaluated by regression and the models were chosen based on the analysis of identity. The neutral detergent

  14. Neuroprotective mechanism of Kai Xin San: upregulation of hippocampal insulin-degrading enzyme protein expression and acceleration of amyloid-beta degradation

    Directory of Open Access Journals (Sweden)

    Na Wang

    2017-01-01

    Full Text Available Kai Xin San is a Chinese herbal formula composed of Radix Ginseng , Poria , Radix Polygalae and Acorus Tatarinowii Rhizome . It has been used in China for many years for treating amnesia. Kai Xin San ameliorates amyloid-β (Aβ-induced cognitive dysfunction and is neuroprotective in vivo , but its precise mechanism remains unclear. Expression of insulin-degrading enzyme (IDE, which degrades Aβ, is strongly correlated with cognitive function. Here, we injected rats with exogenous Aβ42 (200 μM, 5 μL into the hippocampus and subsequently administered Kai Xin San (0.54 or 1.08 g/kg/d intragastrically for 21 consecutive days. Hematoxylin-eosin and Nissl staining revealed that Kai Xin San protected neurons against Aβ-induced damage. Furthermore, enzyme-linked immunosorbent assay, western blot and polymerase chain reaction results showed that Kai Xin San decreased Aβ42 protein levels and increased expression of IDE protein, but not mRNA, in the hippocampus. Our findings reveal that Kai Xin San facilitates hippocampal Aβ degradation and increases IDE expression, which leads, at least in part, to the alleviation of hippocampal neuron injury in rats.

  15. Neuroprotective mechanism of Kai Xin San: upregulation of hippocampal insulin-degrading enzyme protein expression and acceleration of amyloid-beta degradation.

    Science.gov (United States)

    Wang, Na; Jia, Yong-Ming; Zhang, Bo; Xue, Di; Reeju, Maharjan; Li, Yan; Huang, Shu-Ming; Liu, Xue-Wei

    2017-04-01

    Kai Xin San is a Chinese herbal formula composed of Radix Ginseng, Poria, Radix Polygalae and Acorus Tatarinowii Rhizome. It has been used in China for many years for treating amnesia. Kai Xin San ameliorates amyloid-β (Aβ)-induced cognitive dysfunction and is neuroprotective in vivo, but its precise mechanism remains unclear. Expression of insulin-degrading enzyme (IDE), which degrades Aβ, is strongly correlated with cognitive function. Here, we injected rats with exogenous Aβ42 (200 μM, 5 μL) into the hippocampus and subsequently administered Kai Xin San (0.54 or 1.08 g/kg/d) intragastrically for 21 consecutive days. Hematoxylin-eosin and Nissl staining revealed that Kai Xin San protected neurons against Aβ-induced damage. Furthermore, enzyme-linked immunosorbent assay, western blot and polymerase chain reaction results showed that Kai Xin San decreased Aβ42 protein levels and increased expression of IDE protein, but not mRNA, in the hippocampus. Our findings reveal that Kai Xin San facilitates hippocampal Aβ degradation and increases IDE expression, which leads, at least in part, to the alleviation of hippocampal neuron injury in rats.

  16. Substrate recognition in ER-associated degradation mediated by Eps1, a member of the protein disulfide isomerase family

    OpenAIRE

    Wang, Qiongqing; Chang, Amy

    2003-01-01

    Pma1-D378N is a misfolded plasma membrane protein in yeast that is prevented from delivery to the cell surface and targeted instead for ER-associated degradation (ERAD). Degradation of Pma1-D378N is dependent on the ubiquitin ligase Doa10 and the ubiquitin chaperone Cdc48. Recognition of Pma1-D378N by the ERAD pathway is dependent on Eps1, a transmembrane member of the protein disulfide isomerase (PDI) oxidoreductase family. Eps1 has two thioredoxin-like domains containing a CPHC and a CDKC a...

  17. Dominant negative mutant Cyclin T1 proteins inhibit HIV transcription by specifically degrading Tat

    Directory of Open Access Journals (Sweden)

    Okamoto Takashi

    2008-07-01

    Full Text Available Abstract Background The positive transcription elongation factor b (P-TEFb is an essential cellular co-factor for the transcription of the human immunodeficiency virus type 1 (HIV-1. The cyclin T1 (CycT1 subunit of P-TEFb associates with a viral protein, Tat, at the transactivation response element (TAR. This represents a critical and necessary step for the stimulation of transcriptional elongation. Therefore, CycT1 may serve as a potential target for the development of anti-HIV therapies. Results To create effective inhibitors of HIV transcription, mutant CycT1 proteins were constructed based upon sequence similarities between CycT1 and other cyclin molecules, as well as the defined crystal structure of CycT1. One of these mutants, termed CycT1-U7, showed a potent dominant negative effect on Tat-dependent HIV transcription despite a remarkably low steady-state expression level. Surprisingly, the expression levels of Tat proteins co-expressed with CycT1-U7 were significantly lower than Tat co-expressed with wild type CycT1. However, the expression levels of CycT1-U7 and Tat were restored by treatment with proteasome inhibitors. Concomitantly, the dominant negative effect of CycT1-U7 was abolished by these inhibitors. Conclusion These results suggest that CycT1-U7 inhibits HIV transcription by promoting a rapid degradation of Tat. These mutant CycT1 proteins represent a novel class of specific inhibitors for HIV transcription that could potentially be used in the design of anti-viral therapy.

  18. Rumen microbial abundance and fermentation profile during severe subacute ruminal acidosis and its modulation by plant derived alkaloids in vitro.

    Science.gov (United States)

    Mickdam, Elsayed; Khiaosa-Ard, Ratchaneewan; Metzler-Zebeli, Barbara U; Klevenhusen, Fenja; Chizzola, Remigius; Zebeli, Qendrim

    2016-06-01

    Rumen microbiota have important metabolic functions for the host animal. This study aimed at characterizing changes in rumen microbial abundances and fermentation profiles using a severe subacute ruminal acidosis (SARA) in vitro model, and to evaluate a potential modulatory role of plant derived alkaloids (PDA), containing quaternary benzophenanthridine and protopine alkaloids, of which sanguinarine and chelerythrine were the major bioactive compounds. Induction of severe SARA strongly affected the rumen microbial composition and fermentation variables without suppressing the abundance of total bacteria. Protozoa and fungi were more sensitive to the low ruminal pH condition than bacteria. Induction of severe SARA clearly depressed degradation of fiber (P < 0.001), which came along with a decreased relative abundance of fibrolytic Ruminococcus albus and Fibrobacter succinogenes (P < 0.001). Under severe SARA conditions, the genus Prevotella, Lactobacillus group, Megasphaera elsdenii, and Entodinium spp. (P < 0.001) were more abundant, whereas Ruminobacter amylophilus was less abundant. SARA largely suppressed methane formation (-70%, P < 0.001), although total methanogenic 16S rRNA gene abundance was not affected. According to principal component analysis, Methanobrevibacter spp. correlated to methane concentration. Addition of PDA modulated ruminal fermentation under normal conditions such as enhanced (P < 0.05) concentration of total SCFA, propionate and valerate, and increased (P < 0.05) degradation of crude protein compared with the unsupplemented control diet. Our results indicate strong shifts in the microbial community during severe SARA compared to normal conditions. Supplementation of PDA positively modulates ruminal fermentation under normal ruminal pH conditions. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Development and psychometric evaluation of the Utrecht grief rumination scale

    NARCIS (Netherlands)

    Eisma, Maarten C.; Stroebe, Margaret; Schut, Henk A W; Van Den Bout, Jan; Boelen, Paul A.; Stroebe, Wolfgang

    2014-01-01

    Two different types of ruminative coping, depressive rumination and grief rumination, negatively influence bereavement outcome. Although grief-specific rumination is likely to be relevant in the bereavement context no internationally validated scale to measure grief rumination exists. Therefore, the

  20. Memory formation for trace fear conditioning requires ubiquitin-proteasome mediated protein degradation in the prefrontal cortex.

    Directory of Open Access Journals (Sweden)

    David S Reis

    2013-10-01

    Full Text Available The cellular mechanisms supporting plasticity during memory consolidation have been a subject of considerable interest. De novo protein and mRNA synthesis in several brain areas are critical, and more recently protein degradation, mediated by the ubiquitin-proteasome system (UPS, has been shown to be important. Previous work clearly establishes a relationship between protein synthesis and protein degradation in the amygdala, but it is unclear whether cortical mechanisms of memory consolidation are similar to those in the amygdala. Recent work demonstrating a critical role for prefrontal cortex (PFC in the acquisition and consolidation of fear memory allows us to address this question. Here we use a PFC-dependent fear conditioning protocol to determine whether UPS mediated protein degradation is necessary for memory consolidation in PFC. Groups of rats were trained with auditory delay or trace fear conditioning and sacrificed 60 min after training. PFC tissue was then analyzed to quantify the amount of polyubiquinated protein. Other animals were trained with similar procedures but were infused with either a proteasome inhibitor (clasto-lactacystin β-lactone or a translation inhibitor (anisomycin in the PFC immediately after training. Our results show increased UPS-mediated protein degradation in the PFC following trace but not delay fear conditioning. Additionally, post-training proteasome or translation inhibition significantly impaired trace but not delay fear memory when tested the next day. Our results further support the idea that the PFC is critical for trace but not delay fear conditioning highlight the role of UPS-mediated degradation as critical for synaptic plasticity.

  1. Complete genome sequence of the anaerobic, protein-degrading hyperthermophilic crenarchaeon Desulfurococcus kamchatkensis.

    Science.gov (United States)

    Ravin, Nikolai V; Mardanov, Andrey V; Beletsky, Alexey V; Kublanov, Ilya V; Kolganova, Tatiana V; Lebedinsky, Alexander V; Chernyh, Nikolai A; Bonch-Osmolovskaya, Elizaveta A; Skryabin, Konstantin G

    2009-04-01

    Desulfurococcus kamchatkensis is an anaerobic organotrophic hyperthermophilic crenarchaeon isolated from a terrestrial hot spring. Its genome consists of a single circular chromosome of 1,365,223 bp with no extrachromosomal elements. A total of 1,474 protein-encoding genes were annotated, among which 205 are exclusive for D. kamchatkensis. The search for a replication origin site revealed a single region coinciding with a global extreme of the nucleotide composition disparity curve and containing a set of crenarchaeon-type origin recognition boxes. Unlike in most archaea, two genes encoding homologs of the eukaryotic initiator proteins Orc1 and Cdc6 are located distantly from this site. A number of mobile elements are present in the genome, including seven transposons representing IS607 and IS200/IS605 families and multiple copies of miniature inverted repeat transposable elements. Two large clusters of regularly interspaced repeats are present; none of the spacer sequences matches known archaeal extrachromosomal elements, except one spacer matches the sequence of a resident gene of D. kamchatkensis. Many of the predicted metabolic enzymes are associated with the fermentation of peptides and sugars, including more than 30 peptidases with diverse specificities, a number of polysaccharide degradation enzymes, and many transporters. Consistently, the genome encodes both enzymes of the modified Embden-Meyerhof pathway of glucose oxidation and a set of enzymes needed for gluconeogenesis. The genome structure and content reflect the organism's nutritionally diverse, competitive natural environment, which is periodically invaded by viruses and other mobile elements.

  2. The interplay of Hrd3 and the molecular chaperone system ensures efficient degradation of malfolded secretory proteins

    Science.gov (United States)

    Mehnert, Martin; Sommermeyer, Franziska; Berger, Maren; Kumar Lakshmipathy, Sathish; Gauss, Robert; Aebi, Markus; Jarosch, Ernst; Sommer, Thomas

    2015-01-01

    Misfolded proteins of the secretory pathway are extracted from the endoplasmic reticulum (ER), polyubiquitylated by a protein complex termed the Hmg-CoA reductase degradation ligase (HRD-ligase), and degraded by cytosolic 26S proteasomes. This process is termed ER-associated protein degradation (ERAD). We previously showed that the membrane protein Der1, which is a subunit of the HRD-ligase, is involved in the export of aberrant polypeptides from the ER. Unexpectedly, we also uncovered a close spatial proximity of Der1 and the substrate receptor Hrd3 in the ER lumen. We report here on a mutant Hrd3KR that is selectively defective for ERAD of soluble proteins. Hrd3KR displays subtle structural changes that affect its positioning toward Der1. Furthermore, increased quantities of the ER-resident Hsp70-type chaperone Kar2 and the Hsp40-type cochaperone Scj1 bind to Hrd3KR. Of note, deletion of SCJ1 impairs ERAD of model substrates and causes the accumulation of client proteins at Hrd3. Our data imply a function of Scj1 in the removal of malfolded proteins from the receptor Hrd3, which facilitates their delivery to downstream-acting components like Der1. PMID:25428985

  3. Effect of Rapid Chilling on Beef Quality and Cytoskeletal Protein Degradation in M. longissimus of Chinese Yellow Crossbred Bulls.

    Science.gov (United States)

    Mao, Yanwei; Zhang, Yimin; Liang, Rongrong; Ren, Lulu; Zhu, He; Li, Ke; Zhu, Lixian; Luo, Xin

    2012-08-01

    The objective of this study was to investigate the effect of rapid chilling (RC) on beef quality and the degradation of cytoskeletal proteins. Twenty Chinese Yellow crossbred bulls were selected and randomly divided into two groups. RC and conventional chilling (CC) were applied to left and right sides of the carcasses respectively after slaughtering. To determine whether electrical stimulation (ES) treatment can alleviate the potential hazard of RC on meat quality, ES was applied to one group. The effects of RC and ES were determined by meat color, shear force and cytoskeletal protein degradation postmortem (PM). The results showed that RC decreased beef tenderness at 1 d and 3 d postmortem, but had no detrimental effect on meat color. Western blotting showed that RC decreased the degradation rate of desmin and troponin-T, but the effects weakened gradually as postmortem aging extended. Degradation rates of both desmin and troponin-T were accelerated by ES. The combination of RC and ES could improve beef color, accelerate degradation rate of cytoskeletal protein and improve beef tenderness.

  4. Inhibition of ABCA1 protein degradation promotes HDL cholesterol efflux capacity and RCT and reduces atherosclerosis in mice.

    Science.gov (United States)

    Huang, LinZhang; Fan, BaoYan; Ma, Ang; Shaul, Philip W; Zhu, HaiBo

    2015-05-01

    ABCA1 plays a key role in the initial lipidation of apoA-I, which generates circulating HDL cholesterol. Whereas it is known that the transcriptional upregulation of ABCA1 promotes HDL formation and reverse cholesterol transport (RCT), it is not known how the inhibition of ABCA1 protein degradation impacts HDL function. Employing the small molecule triacetyl-3-hydroxyphenyladenosine (IMM-H007), we determined how the attenuation of ABCA1 protein degradation affects HDL cholesterol efflux capacity, RCT, and atherosclerotic lesion formation. Pulse-chase analysis revealed that IMM-H007 inhibits ABCA1 degradation and facilitates its cell-surface localization in macrophages, and additional studies in macrophages showed that IMM-H007 thereby promotes cholesterol efflux. IMM-H007 treatment of Paigen diet-fed mice caused an increase in circulating HDL level, it increased the cholesterol efflux capacity of HDL, and it enhanced in vivo RCT from macrophages to the plasma, liver, and feces. Furthermore, ABCA1 degradation suppression by IMM-H007 reduced atherosclerotic plaque formation in apoE(-/-) mice. Thus, via effects on both ABCA1-expressing cells and circulating HDL function, the inhibition of ABCA1 protein degradation by IMM-H007 promotes HDL cholesterol efflux capacity and RCT and attenuates atherogenesis. IMM-H007 potentially represents a lead compound for the development of agents to augment HDL function. Copyright © 2015 by the American Society for Biochemistry and Molecular Biology, Inc.

  5. Effect of Rapid Chilling on Beef Quality and Cytoskeletal Protein Degradation in of Chinese Yellow Crossbred Bulls

    Directory of Open Access Journals (Sweden)

    Yanwei Mao

    2012-08-01

    Full Text Available The objective of this study was to investigate the effect of rapid chilling (RC on beef quality and the degradation of cytoskeletal proteins. Twenty Chinese Yellow crossbred bulls were selected and randomly divided into two groups. RC and conventional chilling (CC were applied to left and right sides of the carcasses respectively after slaughtering. To determine whether electrical stimulation (ES treatment can alleviate the potential hazard of RC on meat quality, ES was applied to one group. The effects of RC and ES were determined by meat color, shear force and cytoskeletal protein degradation postmortem (PM. The results showed that RC decreased beef tenderness at 1 d and 3 d postmortem, but had no detrimental effect on meat color. Western blotting showed that RC decreased the degradation rate of desmin and troponin-T, but the effects weakened gradually as postmortem aging extended. Degradation rates of both desmin and troponin-T were accelerated by ES. The combination of RC and ES could improve beef color, accelerate degradation rate of cytoskeletal protein and improve beef tenderness.

  6. Antimicrobial Effects of Red Clover (Trifolium pratense) Phenolic Extract on the Ruminal Hyper Ammonia-producing Bacterium, Clostridium sticklandii SR

    Science.gov (United States)

    Ruminal proteolysis and subsequent amino acid degradation represent considerable economic loss in ruminant production. The hyper ammonia-producing bacteria (HAB) are responsible for amino acid deamination in the rumen. HAB can be controlled with ionophores, but they are also susceptible to plant-sy...

  7. Plasma urea nitrogen and progesterone concentrations and follicular dynamics in ewes fed proteins of different degradability

    Directory of Open Access Journals (Sweden)

    Gustavo Bianchi Lazarin

    2012-07-01

    Full Text Available The effects of overfeeding with protein of different degradability on body condition, plasma urea nitrogen and progesterone concentrations, ovulation number and follicular dynamics were assessed in Santa Ines ewes. Twelve ewes were assigned to a randomized block design according to body weight and received overfeeding with soybean meal or with corn gluten meal or maintenance diet for 28 days before ovulation and during the next estrous cycle. Blood samples were taken on days 7, 14, 21, and 28 after the beginning of treatments for analysis of plasma urea nitrogen and on days 3, 6, 9, 12, and 15 into the estrous cycle for analysis of plasma urea nitrogen and progesterone. Follicular dynamics was monitored daily by ultrasound during one estrous cycle. Dry matter and crude protein intake, weight gain, plasma urea nitrogen concentration before ovulation, number of ovulations, diameter of the largest follicle of the 1st and of the 2nd waves and the growth rate of the largest follicle of the 1st wave were higher in the ewes that received overfeeding. The growth rate of the largest follicle of the 3rd wave was higher in the ewes fed maintenance diet. The back fat thickness, plasma urea nitrogen before ovulation and progesterone concentrations, diameter of the largest follicle of the 2nd wave and growth rate of the largest follicle of the 3rd wave were higher in ewes that received overfeeding with soybean meal. The growth rate of the largest follicle of the 1st wave was higher in ewes that received overfeeding with corn gluten meal. Overfeeding with protein-rich feeds may increase the ovulation number and with soybean meal, it may be effective in increasing plasma progesterone concentration in ewes.

  8. SUMOylation of GTF2IRD1 regulates protein partner interactions and ubiquitin-mediated degradation.

    Directory of Open Access Journals (Sweden)

    Jocelyn Widagdo

    Full Text Available GTF2IRD1 is one of the genes implicated in Williams-Beuren syndrome, a disease caused by haploinsufficiency of certain dosage-sensitive genes within a hemizygous microdeletion of chromosome 7. GTF2IRD1 is a prime candidate for some of the major features of the disease, presumably caused by abnormally reduced abundance of this putative transcriptional repressor protein. GTF2IRD1 has been shown to interact with the E3 SUMO ligase PIASxβ, but the significance of this relationship is largely unexplored. Here, we demonstrate that GTF2IRD1 can be SUMOylated by the SUMO E2 ligase UBC9 and the level of SUMOylation is enhanced by PIASxβ. A major SUMOylation site was mapped to lysine 495 within a conserved SUMO consensus motif. SUMOylation of GTF2IRD1 alters the affinity of the protein for binding partners that contain SUMO-interacting motifs, including a novel family member of the HDAC repressor complex, ZMYM5, and PIASxβ itself. In addition, we show that GTF2IRD1 is targeted for ubiquitination and proteasomal degradation. Cross regulation by SUMOylation modulates this process, thus potentially regulating the level of GTF2IRD1 protein in the cell. These findings, concerning post-translational control over the activity and stability of GTF2IRD1, together with previous work showing how GTF2IRD1 directly regulates its own transcription levels suggest an evolutionary requirement for fine control over GTF2IRD1 activity in the cell.

  9. Astaxanthin protects against oxidative stress and calcium-induced porcine lens protein degradation.

    Science.gov (United States)

    Wu, Tzu-Hua; Liao, Jiahn-Haur; Hou, Wen-Chi; Huang, Fu-Yung; Maher, Timothy J; Hu, Chao-Chien

    2006-03-22

    Astaxanthin (ASTX), a carotenoid with potent antioxidant properties, exists naturally in various plants, algae, and seafoods. In this study, we investigated the in vitro ability of ASTX to protect porcine lens crystallins from oxidative damage by iron-mediated hydroxyl radicals or by calcium ion-activated protease (calpain), in addition to the possible underlying biochemical mechanisms. ASTX (1 mM) was capable of protecting lens crystallins from being oxidized, as measured by changes in tryptophan fluorescence, in the presence of a Fenton reaction solution containing 0.2 mM Fe2+ and 2 mM H2O2. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis demonstrated that beta(high)-crystallin was the most vulnerable protein under these conditions of free radical exposure. The proteolysis of lens crystallins induced by calcium ion-activated calpain was also inhibited by ASTX (0.03-1 mM) as determined by daily measurement of the light-scattering intensity at 405 nm for five consecutive days. ASTX at 1 mM was as potent as a concentration of 0.1 mM calpain inhibitor E64 in protecting the oxidative damage/hydrolysis of porcine crystallins. At a concentration of 1 mM, ASTX provided better protection than the endogenous antioxidant glutathione in terms of suppressing calcium-induced turbidity of lens proteins. Thin-layer chromatography analysis indicated that ASTX interacted with calcium ions to form complexes, which we believe interfere with the hydrolysis of lens crystallins by calcium-activated calpain. This in vitro study shows that ASTX is capable of protecting porcine lens proteins from oxidative insults and degradation by calcium-induced calpain.

  10. Role of cysteine-protease CGHC motifs of ER-60, a protein disulfide isomerase, in hepatic apolipoprotein B100 degradation.

    Science.gov (United States)

    Rutledge, Angela C; Qiu, Wei; Zhang, Rianna; Urade, Reiko; Adeli, Khosrow

    2013-09-01

    Apolipoprotein B100 (apoB), the structural component of very low density lipoproteins (VLDL), is susceptible to misfolding and subsequent degradation by several intracellular pathways. ER-60, which has been implicated in apoB degradation, is a protein disulfide isomerase (PDI) that forms or rearranges disulfide bonds in substrate proteins and also possesses cysteine protease activity. To determine which ER-60 function is important for apoB degradation, adenoviruses encoding wild-type human ER-60 or a mutant form of human ER-60 (C60A, C409A) that lacked cysteine protease activity were overexpressed in HepG2 cells. Overexpression of wild-type ER-60 in HepG2 cells promoted apoB degradation and impaired apoB secretion, but mutant ER-60 overexpression did not. In McArdle RH-7777 cells, VLDL secretion was markedly inhibited following overexpression of wild-type but not mutant ER-60, an effect that could be blocked by oleate treatment. Mutant ER-60 was not trapped on apoB as it was with the control substrate tapasin, suggesting that ER-60's role in apoB degradation is likely unrelated to its protein disulfide isomerase activity. Thus, ER-60 may participate in apoB degradation by acting as a cysteine protease. We postulate that apoB cleavage by ER-60 within the ER lumen could facilitate proteasomal degradation of the C-terminus of translocationally-arrested apoB. Copyright © 2013 Elsevier Inc. All rights reserved.

  11. Trafficking and degradation pathways in pathogenic conversion of prions and prion-like proteins in neurodegenerative diseases.

    Science.gov (United States)

    Victoria, Guiliana Soraya; Zurzolo, Chiara

    2015-09-02

    Several neurodegenerative diseases such as transmissible spongiform encephalopathies, Alzheimer's and Parkinson's diseases are caused by the conversion of cellular proteins to a pathogenic conformer. Despite differences in the primary structure and subcellular localization of these proteins, which include the prion protein, α-synuclein and amyloid precursor protein (APP), striking similarity has been observed in their ability to seed and convert naïve protein molecules as well as transfer between cells. This review aims to cover what is known about the intracellular trafficking of these proteins as well as their degradation mechanisms and highlight similarities in their movement through the endocytic pathway that could contribute to the pathogenic conversion and seeding of these proteins which underlies the basis of these diseases. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Ubiquitination and degradation of the hominoid-specific oncoprotein TBC1D3 is regulated by protein palmitoylation.

    Science.gov (United States)

    Kong, Chen; Lange, Jeffrey J; Samovski, Dmitri; Su, Xiong; Liu, Jialiu; Sundaresan, Sinju; Stahl, Philip D

    2013-05-03

    Expression of the hominoid-specific oncoprotein TBC1D3 promotes enhanced cell growth and proliferation by increased activation of signal transduction through several growth factors. Recently we documented the role of CUL7 E3 ligase in growth factors-induced ubiquitination and degradation of TBC1D3. Here we expanded our study to discover additional molecular mechanisms that control TBC1D3 protein turnover. We report that TBC1D3 is palmitoylated on two cysteine residues: 318 and 325. The expression of double palmitoylation mutant TBC1D3:C318/325S resulted in protein mislocalization and enhanced growth factors-induced TBC1D3 degradation. Moreover, ubiquitination of TBC1D3 via CUL7 E3 ligase complex was increased by mutating the palmitoylation sites, suggesting that depalmitoylation of TBC1D3 makes the protein more available for ubiquitination and degradation. The results reported here provide novel insights into the molecular mechanisms that govern TBC1D3 protein degradation. Dysregulation of these mechanisms in vivo could potentially result in aberrant TBC1D3 expression and promote oncogenesis. Copyright © 2013 Elsevier Inc. All rights reserved.

  13. Export of a Cysteine-Free Misfolded Secretory Protein from the Endoplasmic Reticulum for Degradation Requires Interaction with Protein Disulfide Isomerase

    Science.gov (United States)

    Gillece, Pauline; Luz, José Manuel; Lennarz, William J.; de la Cruz, Francisco Javier; Römisch, Karin

    1999-01-01

    Protein disulfide isomerase (PDI) interacts with secretory proteins, irrespective of their thiol content, late during translocation into the ER; thus, PDI may be part of the quality control machinery in the ER. We used yeast pdi1 mutants with deletions in the putative peptide binding region of the molecule to investigate its role in the recognition of misfolded secretory proteins in the ER and their export to the cytosol for degradation. Our pdi1 deletion mutants are deficient in the export of a misfolded cysteine-free secretory protein across the ER membrane to the cytosol for degradation, but ER-to-Golgi complex transport of properly folded secretory proteins is only marginally affected. We demonstrate by chemical cross-linking that PDI specifically interacts with the misfolded secretory protein and that mutant forms of PDI have a lower affinity for this protein. In the ER of the pdi1 mutants, a higher proportion of the misfolded secretory protein remains associated with BiP, and in export-deficient sec61 mutants, the misfolded secretory protein remain bounds to PDI. We conclude that the chaperone PDI is part of the quality control machinery in the ER that recognizes terminally misfolded secretory proteins and targets them to the export channel in the ER membrane. PMID:10613903

  14. Degradation of proteins by enzymes exuded by Allium porrum roots - a potentially important strategy for acquiring organic nitrogen by plants.

    Science.gov (United States)

    Adamczyk, Bartosz; Godlewski, Mirosław; Smolander, Aino; Kitunen, Veikko

    2009-10-01

    Nitrogen is one of the crucial elements that regulate plant growth and development. It is well-established that plants can acquire nitrogen from soil in the form of low-molecular-mass compounds, namely nitrate and ammonium, but also as amino acids. Nevertheless, nitrogen in the soil occurs mainly as proteins or proteins complexed with other organic compounds. Proteins are believed not to be available to plants. However, there is increasing evidence to suggest that plants can actively participate in proteolysis by exudation of proteases by roots and can obtain nitrogen from digested proteins. To gain insight into the process of organic nitrogen acquisition from proteins by leek roots (Allium porrum L. cv. Bartek), casein, bovine serum albumin and oxidized B-chain of insulin were used; their degradation products, after exposure to plant culture medium, were studied using liquid chromatography-mass spectrometry (LC-MS). Casein was degraded to a great extent, but the level of degradation of bovine serum albumin and the B-chain of insulin was lower. Proteases exuded by roots cleaved proteins, releasing low-molecular-mass peptides that can be taken up by roots. Various peptide fragments produced by digestion of the oxidized B-chain of insulin suggested that endopeptidase, but also exopeptidase activity was present. After identification, proteases were similar to cysteine protease from Arabidopsis thaliana. In conclusion, proteases exuded by roots may have great potential in the plant nitrogen nutrition.

  15. Rumination following bereavement : An overview

    NARCIS (Netherlands)

    Eisma, Maarten C.; Stroebe, Margaret S.

    2017-01-01

    This contribution provides an overview of rumination (i.e., thinking repetitively and recurrently about negative events and/or negative emotions) in adjustment to bereavement. First, we summarise a growing literature on rumination and mental health outcomes of bereavement. Next, we compare two main

  16. Wogonin inhibits tumor angiogenesis via degradation of HIF-1α protein

    Energy Technology Data Exchange (ETDEWEB)

    Song, Xiuming; Yao, Jing; Wang, Fei; Zhou, Mi; Zhou, Yuxin; Wang, Hu; Wei, Libin; Zhao, Li; Li, Zhiyu; Lu, Na, E-mail: luna555@163.com; Guo, Qinglong, E-mail: anticancer_drug@yahoo.com.cn

    2013-09-01

    Wogonin, a plant-derived flavone, has been shown recently to have antitumor effects. However, the mechanisms that wogonin inhibits tumor angiogenesis are not well known. In this study, we investigated the effects of wogonin on expression of hypoxia-inducible factor-1α (HIF-1α) and secretion of vascular endothelial growth factor (VEGF) in tumor cells. We found that wogonin decreased the expression of HIF-1α by affecting its stability and reduced the secretion of VEGF, which suppressed angiogenesis in cancer. Wogonin promoted the degradation of HIF-1α by increasing its prolyl hydroxylation, which depended on prolyl hydroxylase (PHD) and the von Hippel–Lindau tumor suppressor (VHL). Intriguingly, wogonin impeded the binding between heat-shock protein 90 (Hsp90) and HIF-1α. In addition, wogonin down-regulated the Hsp90 client proteins EGFR, Cdk4 and survivin, but did not affect the level of Hsp90. Wogonin also increased ubiquitination of HIF-1α and promoted its degradation in proteasome. We also found that wogonin could inhibit nuclear translocation of HIF-1α. Electrophoresis mobility shift assay (EMSA) showed that wogonin decreased the binding activity of exogenous consensus DNA oligonucleotide with HIF-1α in nuclear extracts from MCF-7 cells. Chromatin immunoprecipitation (ChIP) assay also revealed that HIF-1α directly binded to endogenous hypoxia-responsive element (HRE) and this binding was significantly decreased in MCF-7 cells treated with wogonin. Preliminary results indicated in vivo activity of wogonin against xenograft-induced angiogenesis in nude mice. Taken together, the results suggested that wogonin was a potent inhibitor of HIF-1α and provided a new insight into the mechanisms of wogonin against cancers. - Highlights: • Wogonin is an all around inhibitor of VEGF signaling. • We firstly demonstrate that wogonin inhibits secretion of VEGF by decreasing HIF-1α. • Wogonin enhances PDH and VHL expression and inhibits Hsp90 function.

  17. Acidose ruminal em caprinos

    OpenAIRE

    Cunha, José Diogo de Oliveira e silva Ribeiro da

    2012-01-01

    Dissertação de Mestrado Integrado em Medicina Veterinária Com este trabalho pretendeu-se fazer uma revisão acerca da acidose ruminal em caprinos, visto haver pouca bibliografia sobre o tema. Este trabalho foi baseado na revisão bibliográfica de artigos científicos e completado através da observação de casos clínicos ocorridos durante o estágio curricular. O objectivo deste estudo foi realizar uma breve revisão da anatomia e fisiologia do tracto gastrointestinal dos caprinos assim com...

  18. Oncogenic human papillomavirus E6 proteins target the discs large tumour suppressor for proteasome-mediated degradation.

    Science.gov (United States)

    Gardiol, D; Kühne, C; Glaunsinger, B; Lee, S S; Javier, R; Banks, L

    1999-09-30

    Previous studies have shown that the oncogenic HPV E6 proteins form a complex with the human homologue of the Drosophila tumour suppressor protein, discs large (Dlg). This is mediated by the carboxy terminus of the E6 proteins and involves recognition of at least one PDZ domain of Dlg. This region of E6 is not conserved amongst E6 proteins from the low risk papillomavirus types and, hence, binding of HPV E6 proteins to Dlg correlates with the oncogenic potential of these viruses. We have performed studies to investigate the consequences of the interaction between E6 and Dlg. Mutational analysis of both the HPV18 E6 and Dlg proteins has further defined the regions of E6 and Dlg necessary for complex formation. Strikingly, co-expression of wild type HPV18 E6 with Dlg in vitro or in vivo results in a dramatic decrease in the amount of Dlg protein, whereas mutants of E6 which fail to complex with Dlg have minimal effect on Dlg protein levels. The oncogenic HPV16 E6 also decreased the Dlg levels, but this was not observed with the low risk HPV11 E6 protein. Moreover, a region within the first 544 amino acids of Dlg containing the three PDZ domains confers susceptibility to E6 mediated degradation. Finally, treatment of cells with a proteasome inhibitor overrides the capacity of E6 to degrade Dlg. These results demonstrate that Dlg is targeted by high risk HPV E6 proteins for proteasome mediated degradation.

  19. Determining nutrients degradation kinetics of chickpea (Cicer arietinum straw using nylon bag technique in sheep

    Directory of Open Access Journals (Sweden)

    A. Mirzaei-Aghsaghali

    2012-05-01

    Full Text Available Straw a by-product from grain legume crops is produced in large quantities in Iran. Straw is constant component of ruminant diets on small holder farms; however, there is little information about its nutritive value. Accordingly experiment was conducted to determine the chemical composition and ruminal organic matter (OM and crude protein (CP degradability of chickpea straw using nylon bags (in situ technique. Replicated samples were incubated at 0, 2, 4, 8, 12, 24, 48 and 72 hours in three rumen canulated Ghezel rams with 50±3 kg body weight. Dry matter (DM, CP, ether extract (EE, OM, crude fiber (CF and nitrogen free extract (NFE content of chickpea straws were 92.2, 6.1, 5.5, 92.0, 34.3 and 46.2%, respectively. The soluble fraction (a of the OM and CP of chickpea straw was 17.5 and 40.8% and potential degradability (a+b of OM and CP was 56.7 and 72.0%, res