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Sample records for rumen microbial protein

  1. Maximizing efficiency of rumen microbial protein production

    PubMed Central

    Hackmann, Timothy J.; Firkins, Jeffrey L.

    2015-01-01

    Rumen microbes produce cellular protein inefficiently partly because they do not direct all ATP toward growth. They direct some ATP toward maintenance functions, as long-recognized, but they also direct ATP toward reserve carbohydrate synthesis and energy spilling (futile cycles that dissipate heat). Rumen microbes expend ATP by vacillating between (1) accumulation of reserve carbohydrate after feeding (during carbohydrate excess) and (2) mobilization of that carbohydrate thereafter (during carbohydrate limitation). Protozoa account for most accumulation of reserve carbohydrate, and in competition experiments, protozoa accumulated nearly 35-fold more reserve carbohydrate than bacteria. Some pure cultures of bacteria spill energy, but only recently have mixed rumen communities been recognized as capable of the same. When these communities were dosed glucose in vitro, energy spilling could account for nearly 40% of heat production. We suspect that cycling of glycogen (a major reserve carbohydrate) is a major mechanism of spilling; such cycling has already been observed in single-species cultures of protozoa and bacteria. Interconversions of short-chain fatty acids (SCFA) may also expend ATP and depress efficiency of microbial protein production. These interconversions may involve extensive cycling of intermediates, such as cycling of acetate during butyrate production in certain butyrivibrios. We speculate this cycling may expend ATP directly or indirectly. By further quantifying the impact of reserve carbohydrate accumulation, energy spilling, and SCFA interconversions on growth efficiency, we can improve prediction of microbial protein production and guide efforts to improve efficiency of microbial protein production in the rumen. PMID:26029197

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

    PubMed

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

    2015-11-01

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

  3. Effects of Forage Sources on Rumen Fermentation Characteristics, Performance, and Microbial Protein Synthesis in Midlactation Cows

    PubMed Central

    Xu, Jun; Hou, Yujie; Yang, Hongbo; Shi, Renhuang; Wu, Caixia; Huo, Yongjiu; Zhao, Guoqi

    2014-01-01

    Eight multiparous Holstein cows (632±12 kg BW; 135±16 DIM) were used in a replicated 4×4 Latin square design to evaluate the effects of forage sources on rumen fermentation characteristics, performance, and microbial protein (MCP) synthesis. The forage portion of the diets contained alfalfa hay (AH), oat hay (OH), Leymus chinensis (LC), or rice straw (RS) as the primary source of fiber. Diets were isonitrogenous and isocaloric, and cows were fed four corn silages based total mixed rations with equivalent nonfiber carbohydrate (NFC) and forage neutral detergent fiber (NDF). Dry matter intake was not affected by the source of dietary forages, ranging from 18.83 to 19.20 kg/d, consequently, milk yield was similar among diets. Because of the numerical differences in milk fat and milk protein concentrations, 4% FCM and ECM yields were unchanged (p>0.05). Mean rumen pH, NH3-N content, and concentrations of volatile fatty acids in the rumen fluid were not affected by the treatments (p>0.05). Dietary treatments did not affect the total tract apparent digestibility of dry matter, organic matter, and crude protein (p>0.05); however, digestibility of NDF and acid detergent fiber in RS diet was higher compared with AH, OH, and LC diets (p<0.05). Total purine derivative excretion was higher in cows fed AH, OH, and LC diets compared with those fed RS diet (p<0.05), consequently, estimated MCP synthesis was 124.35 g/d higher in cows fed AH diet compared with those fed RS diet (p<0.05). The results indicated that cows fed AH, OH, LC, and RS diets with an equivalent forage NDF and NFC have no unfavourable effect on the ruminal fermentation and productive parameters. PMID:25050001

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

    PubMed

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

    2011-12-01

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

  5. Truly Absorbed Microbial Protein Synthesis, Rumen Bypass Protein, Endogenous Protein, and Total Metabolizable Protein from Starchy and Protein-Rich Raw Materials: Model Comparison and Predictions.

    PubMed

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

    2015-07-29

    This study was carried out to measure truly absorbed microbial protein synthesis, rumen bypass protein, and endogenous protein loss, as well as total metabolizable protein, from starchy and protein-rich raw feed materials with model comparisons. Predictions by the DVE2010 system as a more mechanistic model were compared with those of two other models, DVE1994 and NRC-2001, that are frequently used in common international feeding practice. DVE1994 predictions for intestinally digestible rumen undegradable protein (ARUP) for starchy concentrates were higher (27 vs 18 g/kg DM, p < 0.05, SEM = 1.2) than predictions by the NRC-2001, whereas there was no difference in predictions for ARUP from protein concentrates among the three models. DVE2010 and NRC-2001 had highest estimations of intestinally digestible microbial protein for starchy (92 g/kg DM in DVE2010 vs 46 g/kg DM in NRC-2001 and 67 g/kg DM in DVE1994, p < 0.05 SEM = 4) and protein concentrates (69 g/kg DM in NRC-2001 vs 31 g/kg DM in DVE1994 and 49 g/kg DM in DVE2010, p < 0.05 SEM = 4), respectively. Potential protein supplies predicted by tested models from starchy and protein concentrates are widely different, and comparable direct measurements are needed to evaluate the actual ability of different models to predict the potential protein supply to dairy cows from different feedstuffs.

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

    PubMed

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

    2017-08-29

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

  7. Rumen degradable protein supply affects microbial efficiency in continuous culture and growth in steers.

    PubMed

    Brooks, M A; Harvey, R M; Johnson, N F; Kerley, M S

    2012-12-01

    We hypothesized that microbial efficiency and output from fermentation in the rumen would be optimized when peptide supply was balanced with peptide requirement of ruminal microflora. This study was conducted to measure response of varying rumen degradable peptide (RDPep) supply on ruminal fermentation characteristics and steer growth. A continuous culture experiment was conducted with diets formulated to achieve a predicted RDPep balance (RDPep supplied above RDPep required) of -0.30 to 1.45% CP with rumen degradable N (RDN) balance (RDN supplied above RDN required) above dietary ammonia-N requirement of microbes. Two additional treatments had RDPep balances of -0.30 and 0.78% CP with insufficient ammonia-N supply to meet microbial requirements. Single-flow fermenters (N = 24; n = 6) were inoculated with rumen fluid and maintained anaerobically at 39°C with a 0.06 h(-1) dilution rate. Inadequate RDN decreased OM digestion and microbial N flow, and increased rumen undegradable N (P < 0.01). Microbial efficiency decreased in RDN-deficient diets and was greatest when RDPep balance did not excessively exceed microbial requirement of RDPep predicted (P < 0.01). A growth study was conducted with 49 yearling, crossbred, Angus steers (initial BW 370 ± 34 kg). Animals were assigned to 1 of 4 treatment groups by BW and further divided into 3 pens with 4 steers per pen to achieve similar initial pen weights. Treatments consisted of 4 isonitrogenous diets balanced for RDN but varying in predicted RDPep balance (0.55%, -0.02%, -0.25%, and -0.65% CP). Animals were maintained on treatment for 70 d with individual BW taken on d 0, 1, 21, 42, 70, and 71. Final BW decreased linearly with decreasing RDPep (P = 0.05). Average daily gain and G:F displayed a quadratic effect with greater ADG and G:F at greater and lesser RDPep levels (P = 0.02). We concluded that balancing RDPep supply to predicted requirement improved fermentation efficiency and microbial output, which in turn

  8. Rumen microorganisms, methane production, and microbial protein synthesis affected by mangosteen peel powder supplement in lactating dairy cows.

    PubMed

    Polyorach, Sineenart; Wanapat, Metha; Cherdthong, Anusorn; Kang, Sungchhang

    2016-03-01

    Four crossbred dairy cows (50 % Holstein-Friesian × 50 % Thai native), 404 ± 50.0 kg of body weight (4 years old) and 90 ± 5 day in milk with daily milk production of 9 ± 2.0 kg/day, were randomly assigned according to a 4 × 4 Latin square design to study the effect of mangosteen (Garcinia mangostana) peel powder (MSP) supplementation on rumen microorganisms, methane production, and microbial protein synthesis fed concentrate containing yeast fermented cassava chip protein (YEFECAP). The treatments were different levels of MSP supplementation at 0, 100, 200, and 300 g/head/day. Rice straw was used as a roughage source fed ad libitum, and concentrate containing YEFECAP at 200 g/kg concentrate was offered corresponding to concentrate-to-milk-yield ratio at 1:2. A quantitative real-time PCR approach was used to determine the population densities of ruminal microorganisms. The results revealed that supplementation of MSP did not affect on Fibrobactor succinogenes, Ruminococcus flavefaciens, and Ruminococcus albus (P > 0.05). However, total bacteria was linearly increased (P < 0.01) while methanogens and protozoal population were linearly decreased (P < 0.01) with increasing level of MSP supplementation. Increasing level of MSP supplement could decrease rumen methane production from 27.5 to 23.7 mmol/100 ml(3). Furthermore, cows that received MSP at 300 g/head/day had the highest microbial crude protein and efficiency of rumen microbial N synthesis (416.8 g/day and 16.2 g/kg organic matter truly digested in the rumen (OMDR), respectively). In conclusion, supplementation of MSP at 300 g/head/day with YEFECAP as a protein source in the concentrate mixture revealed an enhancement of rumen fermentation and methane reduction in lactating dairy cows.

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

    PubMed

    Nguyen, Thien Truong Giang; Wanapat, Metha; Phesatcha, Kampanat; Kang, Sungchhang

    2017-02-01

    Leucaena leucocephala (Leucaena) is a perennial tropical legume that can be directly grazed or harvested and offered to ruminants as hay, silage, or fresh. However, Leucaena contain phenolic compounds, which are considered anti-nutritional factors as these may reduce intake, digestibility and thus animal performance. Therefore, the objective of this experiment was to determine effects of Leucaena silage (LS) feeding levels on rumen microbial populations, N-balance and microbial protein synthesis in dairy steers. Four, rumen fistulated dairy steers with initial weight of 167±12 kg were randomly assigned to receive dietary treatments according to a 4×4 Latin square design. Treatments were as followings: T1 = untreated rice straw (RS; Control), T2 = 70% RS+30% LS, T3 = 40% RS+60% LS, and T4 = 100% LS. Dairy steers were fed rice straw and LS ad libitum and supplemented with concentrate at 0.2% of body weight/d. Results revealed that the rumen microbial population, especially cellulolytic, proteolytic bacteria and fungal zoospores were enhanced in steers that received 60% of LS (p<0.05), whereas the amylolytic bacteria population was not affected by treatments (p>0.05). Protozoal population was linearly decreased with increasing level of LS (p<0.05). Moreover, N-balance and microbial protein synthesis were enhanced by LS feeding (p<0.05) and were the highest in 60% LS group. Based on this study, it could be concluded that replacement of RS with 60% LS significantly improved microbial population and microbial protein synthesis in diary steers.

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

    PubMed Central

    Nguyen, Thien Truong Giang; Wanapat, Metha; Phesatcha, Kampanat; Kang, Sungchhang

    2017-01-01

    Objective Leucaena leucocephala (Leucaena) is a perennial tropical legume that can be directly grazed or harvested and offered to ruminants as hay, silage, or fresh. However, Leucaena contain phenolic compounds, which are considered anti-nutritional factors as these may reduce intake, digestibility and thus animal performance. Therefore, the objective of this experiment was to determine effects of Leucaena silage (LS) feeding levels on rumen microbial populations, N-balance and microbial protein synthesis in dairy steers. Methods Four, rumen fistulated dairy steers with initial weight of 167±12 kg were randomly assigned to receive dietary treatments according to a 4×4 Latin square design. Treatments were as followings: T1 = untreated rice straw (RS; Control), T2 = 70% RS+30% LS, T3 = 40% RS+60% LS, and T4 = 100% LS. Dairy steers were fed rice straw and LS ad libitum and supplemented with concentrate at 0.2% of body weight/d. Results Results revealed that the rumen microbial population, especially cellulolytic, proteolytic bacteria and fungal zoospores were enhanced in steers that received 60% of LS (p<0.05), whereas the amylolytic bacteria population was not affected by treatments (p>0.05). Protozoal population was linearly decreased with increasing level of LS (p<0.05). Moreover, N-balance and microbial protein synthesis were enhanced by LS feeding (p<0.05) and were the highest in 60% LS group. Conclusion Based on this study, it could be concluded that replacement of RS with 60% LS significantly improved microbial population and microbial protein synthesis in diary steers. PMID:27165024

  11. Effects of dietary forage sources on rumen microbial protein synthesis and milk performance in early lactating dairy cows.

    PubMed

    Zhu, W; Fu, Y; Wang, B; Wang, C; Ye, J A; Wu, Y M; Liu, J-X

    2013-03-01

    The objective of this study was to evaluate the effects of dietary forage sources on milk performance, rumen microbial protein synthesis, and N utilization in early lactation dairy cows. Twelve primiparous Chinese Holstein dairy cows (45 ± 6.0 DIM) were used in a 3 × 3 Latin square design. Diets were isonitrogenous and isocaloric, with a forage-to-concentrate ratio of 45:55 [dry matter (DM) basis] and contained similar concentrate mixtures. Different forage sources were then added (on a DM basis): 21% corn silage, 19% corn stover, and 5% alfalfa hay (CS); 19% corn silage, 21% Chinese wild rye hay and 5% alfalfa hay (CWR); or 19% corn silage, 9% Chinese wild rye hay, and 17% alfalfa hay (AH). Each period lasted for 21 d, with the first 14 d for an adaptation period. Dry matter intake was not affected by the source of dietary forage. Milk yield was higher for cows fed AH than those fed CS, with an intermediate value for CWR. Milk protein content was higher in the cows fed AH compared with CWR (3.02 vs. 2.92%), with CS (2.95%) at an intermediate position. The contents of milk fat and lactose were not different among the treatments. However, milk efficiency (milk yield/DM intake) was higher for cows fed AH than those fed CS, with those fed CWR intermediate. Cows fed AH had higher microbial protein yield and metabolizable protein than those fed CS or CWR. The concentrations of urea N in the urine, blood, and milk were decreased for cows fed AH, indicating an increased N conversion. The results indicated that corn stover could replace Chinese wild rye grass in the diets for lactating cows and that a high proportion of alfalfa hay in the diet is beneficial for milk protein production by increasing microbial protein yield. This can be attributed to the improving the supply of rumen-available energy.

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

    PubMed

    McSweeney, C S; Denman, S E

    2007-11-01

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

  13. Replacing alfalfa hay with dry corn gluten feed and Chinese wild rye grass: Effects on rumen fermentation, rumen microbial protein synthesis, and lactation performance in lactating dairy cows.

    PubMed

    Hao, X Y; Gao, H; Wang, X Y; Zhang, G N; Zhang, Y G

    2017-04-01

    This experiment was conducted to investigate nutrient digestibility, rumen microbial protein synthesis, and lactation performance when a portion of alfalfa was replaced with combinations of dry corn gluten feed (DCGF) and Chinese wild rye grass in the diet of lactating cows. Six multiparous and 3 primiparous Chinese Holsteins were arranged in a replicated 3 × 3 Latin square experiment for 21-d periods. The animals were fed 1 of 3 treatment diets during each period: (1) 0% DCGF (0DCGF); (2) 6.5% DCGF (7DCGF); and (3) 11% DCGF (11DCGF). Diets were isonitrogenous, and a portion of alfalfa hay was replaced with DCGF and Chinese wild rye grass, with similar concentrate mixtures and corn silage contents. The dry matter intake was greater for 11DCGF (21.9 kg/d) than for 0DCGF (20.7 kg/d) or 7DCGF (21.2 kg/d). The treatment diets did not result in difference in milk production, fat and lactose concentration, or yield. Compared with 0DCGF, the ration containing 11% DCGF improved the milk protein concentration. Dry matter and neutral detergent fiber digestibility was greater for 7DCGF (62.7% and 45.6%) and 11DCGF (63.1% and 47.2%) than for 0DCGF (59.4% and 42.3%), and the nitrogen digestibility was similar for the 3 treatments. The concentration of rumen volatile fatty acids was higher in cows fed the 11DCGF diet than in those fed the 0DCGF diet, with no difference between the 7DCGF and 11DCGF diets. The estimated microbial crude protein yield was greater for the 11DCGF diet (1985.1 g/d) than for the 0DCGF diet (1745.0 g/d), with no difference between the 0DCGF and 7DCGF diets. Thus, it appears that feeding DCGF and Chinese wild rye grass in combination can effectively replace a portion of alfalfa hay in the rations of lactating dairy cows.

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

    PubMed

    Polyorach, S; Wanapat, M

    2015-06-01

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

  15. Effect of dietary energy source and level on nutrient digestibility, rumen microbial protein synthesis, and milk performance in lactating dairy cows.

    PubMed

    Zhou, X Q; Zhang, Y D; Zhao, M; Zhang, T; Zhu, D; Bu, D P; Wang, J Q

    2015-10-01

    This study was conducted to examine the effects of dietary energy source and level on intake, digestion, rumen microbial protein synthesis, and milk production in lactating dairy cows, using corn stover as a forage source. Eight multiparous Holstein cows, 4 of which were fitted with rumen cannulas, were evaluated in a replicated 4 × 4 Latin square design, with each period lasting 21 d. The cows were randomly assigned into 4 treatment groups: low-energy (LE) ground corn (GC), LE steam-flaked corn (SFC), high-energy (HE) GC, and HE SFC. Changes to ruminal energy degradation rates were induced by feeding the cows diets of either finely ground corn or SFC as components of diets with the same total energy level. Milk yield, milk protein content and yield, and milk lactose yield all increased in response to higher levels of dietary energy, whereas contents of milk fat and lactose were unaffected. Cows fed HE diets had a higher crude microbial protein yield and total-tract apparent digestibility than those receiving LE diets. Milk yield, milk protein yield, and microbial protein yield were also higher when SFC replaced GC as the main energy source for lactating cows fed LE diets. These results suggest that an increased dietary energy level and ruminal degradation rate are beneficial to milk protein production, which we suggest is due to increased yields of microbial proteins, when cows are fed corn stover as a dietary forage source.

  16. Effect of Fusarium toxin-contaminated triticale and forage-to-concentrate ratio on fermentation and microbial protein synthesis in the rumen.

    PubMed

    Hildebrand, B; Boguhn, J; Dänicke, S; Rodehutscord, M

    2012-04-01

    In this study, the effect of Fusarium toxin-contaminated triticale (FUS) at high (60%) and low (30%) concentrate proportion in ruminant rations on ruminal fermentation, microbial protein synthesis and digestibility was investigated, using in vivo and in vitro methods. Significant effects of the forage-to-concentrate ratio on ruminal degradation and digestibility of crude nutrients and detergent fibre fractions as well as on the pH value and the concentration of short chain fatty acids (SCFA) in rumen fluid were found. The production of SCFA was affected, and the degradation of crude fibre and neutral detergent fibre in the rumen was reduced by the inclusion of FUS at high concentrate proportion. The efficiency of microbial crude protein synthesis was higher in diets with 60% than in diets with 30% concentrates, but was impaired in the presence of FUS in vitro at the high concentrate level. Marginal effects of FUS on the amino acid pattern of microbial protein were detected. It was concluded that the use of FUS in high concentrate diets can influence ruminal fermentation and microbial protein synthesis at a dietary deoxynivalenol concentration below 5 mg/kg dry matter. © 2011 Blackwell Verlag GmbH.

  17. Rumen microbes and microbial protein synthesis in Thai native beef cattle fed with feed blocks supplemented with a urea-calcium sulphate mixture.

    PubMed

    Cherdthong, Anusorn; Wanapat, Metha

    2013-12-01

    The influence of slow-release urea ( urea-calcium sulphate mixture; U-CaS) in feed blocks on rumen micro-organisms, predominant cellulolytic bacteria, microbial protein synthesis and ecology was studied in Thai native beef cattle. Four animals with an initial body weight of 100 ± 3.0 kg were randomly assigned to a 4 × 4 Latin square design with four dietary treatments (U-CaS in iso-nitrogen feed blocks at 0, 120, 150 and 180 g/kg dry matter (DM), respectively; U-CaS replaced urea). After 21 days of experimental feeding, rumen fluid was collected at 0 and 4 h after feeding. The mean intake of feed blocks and other feedstuffs offered (rice straw and concentrates) amounted to 0.3, 2.3 and 0.6 kg DM/day, respectively. Inclusion of U-CaS did not altered pH and temperature in the rumen. However, ruminal NH3-N concentration decreased quadratically (p < 0.05) in response to U-CaS inclusion, with the lowest value at 180 g U-CaS per kg feed block. With inclusion of U-CaS, the populations of rumen bacteria increased quadratically (p < 0.05) and counts of fungal zoospores were linearly enhanced (p < 0.05), being highest at 180 g U-CaS per kg feed block. Supplementation of U-CaS increased the concentration of total bacteria linearly (p < 0.05) and of Fibrobacter succinogenes quadratically (p < 0.05), whereas Ruminococcus flavefaciens and Ruminococcus albus were not affected by dietary treatments. Microbial crude protein yield and efficiency of microbial nitrogen (N) synthesis were linearly increased with different levels of U-CaS addition. Furthermore, current data clearly indicate that inclusion of U-CaS in feed blocks can affect micro-organism diversity and major cellulolytic bacteria.

  18. Effects of Physical Form and Urea Treatment of Rice Straw on Rumen Fermentation, Microbial Protein Synthesis and Nutrient Digestibility in Dairy Steers

    PubMed Central

    Gunun, P.; Wanapat, M.; Anantasook, N.

    2013-01-01

    This study was designed to determine the effect of physical form and urea treatment of rice straw on rumen fermentation, microbial protein synthesis and nutrient digestibility. Four rumen-fistulated dairy steers were randomly assigned according to a 2 (2 factorial arrangement in a 4 (4 Latin square design to receive four dietary treatments. Factor A was roughage source: untreated rice straw (RS) and urea-treated (3%) rice straw (UTRS), and factor B was type of physical form of rice straw: long form rice straw (LFR) and chopped (4 cm) rice straw (CHR). The steers were offered the concentrate at 0.5% body weight (BW) /d and rice straw was fed ad libitum. DM intake and nutrient digestibility were increased (p<0.05) by urea treatment. Ruminal pH were decreased (p<0.05) in UTRS fed group, while ruminal ammonia nitrogen (NH3-N) and blood urea nitrogen (BUN) were increased (p<0.01) by urea treatment. Total volatile fatty acid (VFA) concentrations increased (p<0.01) when steers were fed UTRS. Furthermore, VFA concentrations were not altered by treatments (p>0.05), except propionic acid (C3) was increased (p<0.05) in UTRS fed group. Nitrogen (N) balance was affected by urea treatment (p<0.05). Microbial protein synthesis (MCP) synthesis were greater by UTRS and CHR group (p<0.05). The efficiency of microbial N synthesis was greater for UTRS than for RS (p<0.05). From these results, it can be concluded that using the long form combined with urea treatment of rice straw improved feed intake, digestibility, rumen fermentation and efficiency of microbial N synthesis in crossbred dairy steers. PMID:25049759

  19. Effects of physical form and urea treatment of rice straw on rumen fermentation, microbial protein synthesis and nutrient digestibility in dairy steers.

    PubMed

    Gunun, P; Wanapat, M; Anantasook, N

    2013-12-01

    This study was designed to determine the effect of physical form and urea treatment of rice straw on rumen fermentation, microbial protein synthesis and nutrient digestibility. Four rumen-fistulated dairy steers were randomly assigned according to a 2 (2 factorial arrangement in a 4 (4 Latin square design to receive four dietary treatments. Factor A was roughage source: untreated rice straw (RS) and urea-treated (3%) rice straw (UTRS), and factor B was type of physical form of rice straw: long form rice straw (LFR) and chopped (4 cm) rice straw (CHR). The steers were offered the concentrate at 0.5% body weight (BW) /d and rice straw was fed ad libitum. DM intake and nutrient digestibility were increased (p<0.05) by urea treatment. Ruminal pH were decreased (p<0.05) in UTRS fed group, while ruminal ammonia nitrogen (NH3-N) and blood urea nitrogen (BUN) were increased (p<0.01) by urea treatment. Total volatile fatty acid (VFA) concentrations increased (p<0.01) when steers were fed UTRS. Furthermore, VFA concentrations were not altered by treatments (p>0.05), except propionic acid (C3) was increased (p<0.05) in UTRS fed group. Nitrogen (N) balance was affected by urea treatment (p<0.05). Microbial protein synthesis (MCP) synthesis were greater by UTRS and CHR group (p<0.05). The efficiency of microbial N synthesis was greater for UTRS than for RS (p<0.05). From these results, it can be concluded that using the long form combined with urea treatment of rice straw improved feed intake, digestibility, rumen fermentation and efficiency of microbial N synthesis in crossbred dairy steers.

  20. Effect of Grape Pomace Powder, Mangosteen Peel Powder and Monensin on Nutrient Digestibility, Rumen Fermentation, Nitrogen Balance and Microbial Protein Synthesis in Dairy Steers.

    PubMed

    Foiklang, S; Wanapat, M; Norrapoke, T

    2016-10-01

    This study was designed to investigate the effect of grape pomace powder (GPP), mangosteen peel powder (MPP) and monensin on feed intake, nutrients digestibility, microorganisms, rumen fermentation characteristic, microbial protein synthesis and nitrogen balance in dairy steers. Four, rumen fistulated dairy steers with initial body weight (BW) of 220±15 kg were randomly assigned according to a 4×4 Latin square design to receive four treatments. The treatments were as follows: T1 = control, T2 = supplementation with monensin at 33 mg/kg diet, T3 = supplementation with GPP at 2% of dry matter intake, and T4 = supplementation with MPP at 30 g/kg diet. The steers were offered the concentrate diet at 0.2% BW and 3% urea treated rice straw (UTRS) was fed ad libitum. It was found that GPP supplemented group had higher UTRS intake and nutrient digestibility in terms of neutral detergent fiber and acid detergent fiber than those in control group (p<0.05). Ammonia nitrogen (NH3-N) and blood urea-nitrogen concentration were higher in monensin, GPP and MPP supplemented groups (p<0.05). Total volatile fatty acids and propionate in the GPP group were higher than those in the control group (p<0.05) while acetate concentration, and acetate to propionate ratio were decreased (p<0.01) when steers were supplemented with GPP, monensin, and MPP, respectively. Moreover, protozoal populations in GPP, MPP, and monensin supplementation were significantly lower than those in the control group (p<0.05), while cellulolytic bacterial population was significantly higher in the control group (p<0.05). Nitrogen retention, microbial crude protein and efficiency of microbial nitrogen synthesis were found significantly higher in steers that received GPP (p<0.05). Based on this study it could be concluded that the GPP has potential as an alternative feed supplement in concentrate diets which can result in improved rumen fermentation efficiency, digestibility and microbial protein synthesis in

  1. Effect of Grape Pomace Powder, Mangosteen Peel Powder and Monensin on Nutrient Digestibility, Rumen Fermentation, Nitrogen Balance and Microbial Protein Synthesis in Dairy Steers

    PubMed Central

    Foiklang, S.; Wanapat, M.; Norrapoke, T.

    2016-01-01

    This study was designed to investigate the effect of grape pomace powder (GPP), mangosteen peel powder (MPP) and monensin on feed intake, nutrients digestibility, microorganisms, rumen fermentation characteristic, microbial protein synthesis and nitrogen balance in dairy steers. Four, rumen fistulated dairy steers with initial body weight (BW) of 220±15 kg were randomly assigned according to a 4×4 Latin square design to receive four treatments. The treatments were as follows: T1 = control, T2 = supplementation with monensin at 33 mg/kg diet, T3 = supplementation with GPP at 2% of dry matter intake, and T4 = supplementation with MPP at 30 g/kg diet. The steers were offered the concentrate diet at 0.2% BW and 3% urea treated rice straw (UTRS) was fed ad libitum. It was found that GPP supplemented group had higher UTRS intake and nutrient digestibility in terms of neutral detergent fiber and acid detergent fiber than those in control group (p<0.05). Ammonia nitrogen (NH3-N) and blood urea-nitrogen concentration were higher in monensin, GPP and MPP supplemented groups (p<0.05). Total volatile fatty acids and propionate in the GPP group were higher than those in the control group (p<0.05) while acetate concentration, and acetate to propionate ratio were decreased (p<0.01) when steers were supplemented with GPP, monensin, and MPP, respectively. Moreover, protozoal populations in GPP, MPP, and monensin supplementation were significantly lower than those in the control group (p<0.05), while cellulolytic bacterial population was significantly higher in the control group (p<0.05). Nitrogen retention, microbial crude protein and efficiency of microbial nitrogen synthesis were found significantly higher in steers that received GPP (p<0.05). Based on this study it could be concluded that the GPP has potential as an alternative feed supplement in concentrate diets which can result in improved rumen fermentation efficiency, digestibility and microbial protein synthesis in

  2. Effect of carbohydrate sources and levels of cotton seed meal in concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in young dairy bulls.

    PubMed

    Wanapat, M; Anantasook, N; Rowlinson, P; Pilajun, R; Gunun, P

    2013-04-01

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

  3. Rumen microbial communities influence metabolic phenotypes in lambs

    PubMed Central

    Morgavi, Diego P.; Rathahao-Paris, Estelle; Popova, Milka; Boccard, Julien; Nielsen, Kristian F.; Boudra, Hamid

    2015-01-01

    The rumen microbiota is an essential part of ruminants shaping their nutrition and health. Despite its importance, it is not fully understood how various groups of rumen microbes affect host-microbe relationships and functions. The aim of the study was to simultaneously explore the rumen microbiota and the metabolic phenotype of lambs for identifying host-microbe associations and potential biomarkers of digestive functions. Twin lambs, separated in two groups after birth were exposed to practices (isolation and gavage with rumen fluid with protozoa or protozoa-depleted) that differentially restricted the acquisition of microbes. Rumen microbiota, fermentation parameters, digestibility and growth were monitored for up to 31 weeks of age. Microbiota assembled in isolation from other ruminants lacked protozoa and had low bacterial and archaeal diversity whereas digestibility was not affected. Exposure to adult sheep microbiota increased bacterial and archaeal diversity independently of protozoa presence. For archaea, Methanomassiliicoccales displaced Methanosphaera. Notwithstanding, protozoa induced differences in functional traits such as digestibility and significantly shaped bacterial community structure, notably Ruminococcaceae and Lachnospiraceae lower up to 6 folds, Prevotellaceae lower by ~40%, and Clostridiaceae and Veillonellaceae higher up to 10 folds compared to microbiota without protozoa. An orthogonal partial least squares-discriminant analysis of urinary metabolome matched differences in microbiota structure. Discriminant metabolites were mainly involved in amino acids and protein metabolic pathways while a negative interaction was observed between methylotrophic methanogens Methanomassiliicoccales and trimethylamine N-oxide. These results stress the influence of gut microbes on animal phenotype and show the potential of metabolomics for monitoring rumen microbial functions. PMID:26528248

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

    PubMed

    Wickersham, T A; Titgemeyer, E C; Cochran, R C; Wickersham, E E; Moore, E S

    2008-11-01

    We evaluated the effect of frequency and amount of rumen-degradable intake protein (DIP) on urea kinetics in steers consuming prairie hay. Five ruminally and duodenally fistulated steers (366 kg of BW) were used in a 5 x 5 Latin square and provided ad libitum access to low-quality prairie hay (4.7% CP). Casein was provided daily in amounts of 61 and 183 mg of N/kg of BW (61/d and 183/d) and every third day in amounts of 61, 183, and 549 mg of N/kg of BW per supplementation event (61/3d, 183/3d, and 549/3d). Periods were 18-d long with 9 d for adaptation and 9 d for collection. Steers were in metabolism crates for total collection of urine and feces. Jugular infusion of (15)N(15)N-urea followed by determination of urinary enrichment of (15)N(15)N-urea and (14)N(15)N-urea was used to determine urea kinetics. Treatment means were separated to evaluate the effects of increasing DIP supplementation and the effects of frequency at the low (61/d vs. 183/3d) and at the high (183/d vs. 549/3d) amounts of DIP provision. Forage OM and total digestible OM intakes were linearly (P < or = 0.05) increased by increasing DIP provision but were not affected by frequency of supplementation at either the low or high amounts. Production and gut entry of urea linearly (P < or = 0.006) increased with DIP provision and tended to be greater (P < or = 0.07) for 549/3d than 183/d but were not different between 61/d and 183/3d. Microbial N flow to the duodenum was linearly (P < 0.001) increased by increasing DIP provision. Additionally, 183/d resulted in greater (P = 0.05) microbial N flow than 549/3d. Incorporation of recycled urea-N into microbial N linearly (P = 0.04) increased with increasing DIP. Microbial incorporation of recycled urea-N was greater for 549/3d than 183/d, with 42 and 23% of microbial N coming from recycled urea-N, respectively. In contrast, there was no difference due to frequency in the incorporation of recycled urea-N by ruminal microbes at the low level of

  5. Effects of an exogenous enzyme preparation on microbial protein synthesis, enzyme activity and attachment to feed in the Rumen Simulation Technique (Rusitec).

    PubMed

    Wang, Y; McAllister, T A; Rode, L M; Beauchemin, K A; Morgavi, D P; Nsereko, V L; Iwaasa, A D; Yang, W

    2001-03-01

    The effects of an exogenous enzyme preparation, the application method and feed type on ruminal fermentation and microbial protein synthesis were investigated using the rumen simulation technique (Rusitec). Steam-rolled barley grain and chopped alfalfa hay were sprayed with water (control, C), an enzyme preparation with a predominant xylanase activity (EF), or autoclaved enzyme (AEF) 24 h prior to feeding, or the enzyme was supplied in the buffer infused into the Rusitec (EI). Microbial N incorporation was measured using (15NH4)2SO4 in the buffer. Spent feed bags were pummelled mechanically in buffer to segregate the feed particle-associated (FPA) and feed particle-bound (FPB) bacterial fractions. Enzymes applied to feed reduced neutral-detergent fibre content, and increased the concentration of reducing sugars in barley grain, but not alfalfa hay. Ruminal cellulolytic bacteria were more numerous with EF than with C. Disappearance of DM from barley grain was higher with EF than with C, but alfalfa was unaffected by EF. Treatment EF increased incorporation of 15N into FPA and FPB fractions at 24 and 48 h. In contrast, AEF reduced the 24 h values, relative to C; AEF and C were similar at 48 h. Infused enzyme (EI) did not affect 15N incorporation. Xylanase activity in effluent was increased by EF and EI, compared to C, but not by AEF. Xylanase activity in FPA was higher at 48 h than at 24 h with all treatments; it was higher with EF than C at 24 and 48 h, but was not altered by AEF or EI. Applying enzymes onto feeds before feeding was more effective than dosing directly into the artificial rumen for increasing ruminal fibrolytic activity.

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

    PubMed

    Hildebrand, Bastian; Boguhn, Jeannette; Rodehutscord, Markus

    2011-10-01

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

  7. Effects of various plant protein sources in high-quality feed block on feed intake, rumen fermentation, and microbial population in swamp buffalo.

    PubMed

    Foiklang, Suban; Wanapat, Metha; Toburan, Wetchasit

    2011-12-01

    This study was designed to determine effect of various plant protein sources in high-quality feed block (HQFB) on feed intake, rumen fermentation, and microbial population in swamp buffalo. Four rumen-fistulated swamp buffaloes (Bubalus bubalis) were randomly assigned according to a 4 × 4 Latin square design. Four kinds of plant protein sources (coarse rice bran (CRB), cassava hay (CH), Phaseolus calcaratus hay, and mulberry hay (MH)) were mixed in the HQFB. HQFBs were allowed to be licked at free choice, and urea-lime-treated rice straw (ULRS) were fed ad libitum. It was found that bacterial population and fungal zoospores in CH-fed group tended to be higher than those in other groups. Moreover, protozoal population in CH, P. calcaratus hay, and MH were lower than those in CRB supplemented group (P < 0.05). Cellulolytic bacterial population was highest in CH-fed group while proteolytic bacteria population was highest in P. calcaratus hay-fed group (P < 0.05). CH-fed group had higher ULRS intake than those in other groups (P < 0.05). Nutrient digestibility of CP, NDF, and ADF in CH-fed group was significantly higher than those in other groups (P < 0.05). Total VFA was highest in CH-fed group (P < 0.05). N absorption was highest in CH-fed group (P < 0.05). Based on this study, it could be concluded that cassava hay, P. calcaratus hay, and mulberry hay are potential to be used as protein sources in the HQFBs especially cassava hay.

  8. Rumen fermentation, microbial protein synthesis, and nutrient flow to the omasum in cattle offered corn silage, grass silage, or whole-crop wheat.

    PubMed

    Owens, D; McGee, M; Boland, T; O'Kiely, P

    2009-02-01

    The objectives of this study were to determine the relative effect of feeding corn silage (CS), fermented whole-crop wheat (FWCW), and urea-treated processed whole-crop wheat (UPWCW) compared with grass silage (GS), each supplemented with concentrates, on forage intake, ruminal fermentation, microbial protein synthesis, some plasma metabolites, and ruminal and total tract digestibility in cattle. Four ruminally fistulated steers with a mean BW of 509 kg (SD 6.3) were used in a 4 x 4 Latin square-designed experiment with each period lasting 21 d. The omasal sampling technique in combination with a triple marker method was used to measure nutrient flows to the omasum with Co-EDTA, Yb acetate, and indigestible NDF as liquid, small particle, and large particle phase markers, respectively. Microbial N flow was assessed from purine base concentrations. Steers fed CS, FWCW, and UPWCW consumed 2.7, 2.4, and 2.6 kg/d more (P < 0.05) forage and total DMI, respectively, than those fed GS-based diets. Rumen pH (P = 0.07) and lactic acid (P = 0.11) concentration did not differ between the forages. Rumen concentration of NH(3)-N was greatest for UPWCW and least for CS (P < 0.001). Total VFA concentrations were greater (P < 0.05) for CS than GS and UPWCW, with FWCW being intermediate. Acetate-to-propionate ratio (P < 0.05) was greater (P < 0.05) for UPWCW than the other forages, which did not differ. Apparent ruminal digestion of OM (P < 0.05) was less for CS, FWCW, and UPWCW than GS. Ruminal NDF digestibility was greater (P < 0.01) for GS than the other forages, which did not differ (P > or = 0.06). Total tract NDF digestibility was less (P < 0.05) for UPWCW than the other forages, with GS being greatest and CS and FWCW being intermediate. Starch intake was less (P < 0.001) for GS than the other forages, but there was no effect of forage on omasal starch flow (P = 0.23) or ruminal digestibility (P = 0.88). Flow of non-NH(3)-N and microbial N was greater (P < 0.05) for CS, FWCW

  9. Effects of quebracho tannin extract (Schinopsis balansae Engl.) and activated charcoal on nitrogen balance, rumen microbial protein synthesis and faecal composition of growing Boer goats.

    PubMed

    Al-Kindi, Amal; Dickhoefer, Uta; Schlecht, Eva; Sundrum, Albert; Schiborra, Anne

    2016-08-01

    Under irrigated arid conditions, organic fertiliser rich in slowly decomposable nitrogen (N) and carbon (C) is needed for soil fertility maintenance. Feeding ruminants with condensed tannins will lower ruminal protein degradation, reduce urinary N excretion and might increase the faecal fraction of slowly decomposable N. Supplementation with activated charcoal (AC) might enrich manure with slowly degrading C. Therefore, we investigated the effects of feeding quebracho tannin extract (QTE) and AC on the N balance of goats, the efficiency of microbial protein synthesis in the rumen (EMPS) and the composition of faeces. The feeding trial comprised three periods; in each period, 12 male Boer goats (28 ± 3.9 kg live weight) were assigned to six treatments: a Control diet (per kg diet 500 g grass hay and 500 g concentrate) and to further five treatments the Control diet was supplemented with QTE (20 g and 40 g/kg; diets QTE2 and QTE4, respectively), with AC (15 g and 30 g/kg, diets AC1.5 and AC3.0, respectively) and a mixture of QTE (20 g/kg) plus AC (15 g/kg) (diet QTEAC). In addition to the N balance, EMPS was calculated from daily excretions of purine derivatives, and the composition of faecal N was determined. There was no effect of QTE and AC supplementation on the intake of organic matter (OM), N and fibre, but apparent total tract digestibility of OM was reduced (p = 0.035). Feeding QTE induced a shift in N excretion from urine to faeces (p ≤ 0.001) without altering N retention. Total N excretion tended to decrease with QTE treatments (p = 0.053), but EMPS was not different between treatments. Faecal C excretion was higher in QTE and AC treatments (p = 0.001) compared with the Control, while the composition of faecal N differed only in concentration of undigested dietary N (p = 0.001). The results demonstrate that QTE can be included into diets of goats up to 40 g/kg, without affecting N utilisation, but simultaneously increasing the

  10. Effects of cold exposure on feed protein degradation, microbial protein synthesis and transfer of plasma urea to the rumen of sheep.

    PubMed

    Kennedy, P M; Christopherson, R J; Milligan, L P

    1982-05-01

    1. Three diets of barley-canola-seed (Brassica campestris), lucerne (Medicago sativa) or chopped brome-grass (Bromus inermis) were given at intervals of 3 h to closely-shorn Suffolk wethers held at a temperature of 1-5 degree (cold) or 22-24 degree (warm). Apparent digestibility of organic matter (OM) and nitrogen was reduced by 0.08-0.05 and 0.04 units respectively for lucerne and brome-grass diets given to cold-exposed sheep, but no treatment effects on digestibility were observed for the barley-CSM diet. Measurements achieved using infusion of the digesta markers 58Co-EDTA and 103Ru-phenanthroline (103Ru-P) showed that cold exposure depressed apparent OM digestion in the stomach and intestines by 33 and 42 g/d for the lucerne diet, and 13 and 35 g/d for the brome-grass diet respectively. 2. The turnover time (h) of the 103Ru-P marker in the rumen of warm sheep was 38.9 for barley-CSM, 18.4 for lucerne, and 15.6 for brome-grass. In cold-exposed sheep, 103Ru-P turnover time (h) tended to be reduced to 32.3, 12.3 and 15.3 for the three diets, respectively. OM fermentation in the stomach was highly related to 103RU-P turnover time for lucerne and brome-grass diets. 3. Cold exposure increased the escape of dietary N from the abomasum by 0.04 and 0.09 of dietary N intake for sheep given lucerne and brome-grass diets respectively. Dietary N degradation was closely related to 103Ru-P turnover time for lucerne, and to the proportion of large particles in rumen digesta for the brome-grass diet. Estimates of feed N degradation made by use of information on the rate of fermentation of the diet in nylon bags and 103Ru-P turnover time were consistently lower than those observed in vivo for barley-CSM and lucerne diets. Intestinal digestibility of non-ammonia N was not significantly changed by cold exposure. 4. Transfer of urea from plasma to the rumen was 1.4-2.5 g N/d for the barley-CSM and lucerne diets, but the value for brome-grass was 4.5-4.9 g N/d. Cold exposure did not

  11. Microbial and chemical composition of liquid-associated bacteria in goats' rumen and fermenters.

    PubMed

    Abecia, L; Soto, E C; Ramos-Morales, E; Molina-Alcaide, E

    2014-10-01

    This study was undertaken to investigate the relationship between chemical composition and microbial profile of rumen liquid-associated bacteria (LAB) in vivo (Murciano-Granadina goats) and in a rumen simulation system (single-flow continuous-culture fermenters). To achieve this aim, analyses of purine bases along with some molecular techniques (quantitative PCR to assess abundance and DGGE to identify biodiversity and bacterial profile) were carried out. A control diet (AHC) based on alfalfa hay (AH) and concentrate (C) in a 1:1 ratio and two experimental diets (AHCBI and AHCBII), in which concentrate was partially replaced with multinutrient blocks, were used. Diets AHCBI and AHCBII included multinutrient blocks differing in the relative amount of two-stage olive cake and the source of protein (sunflower meal vs. fava beans). We aimed to investigate the effect of these blocks on rumen microbiota to evaluate their potential as safe substitutes of cereal-based concentrates. Similar patterns of response to diet were found for chemical composition, microbial abundances and diversity in LAB isolated from goat's rumen and fermenters. Whereas bacterial density (log10 gene copies/g FM: 11.6 and 9.4 for bacteria and methanogens, respectively, in rumen) and diversity indexes (Shannon index: 3.6) were not affected by diet, DGGE analyses showed that bacterial community profile was affected. The cluster analysis suggested differences in bacterial profile between LAB pellets isolated from the rumen of goat and fermenters. A relationship between chemical composition and bacterial community composition in LAB pellets seems to exist. Changes in the former were reflected in the bacterial community profile. Further research is needed to clarify the relationship between chemical and microbial composition of ruminal bacterial pellets with diets of different quality.

  12. The relationship between odd- and branched-chain fatty acids and microbial nucleic acid bases in rumen.

    PubMed

    Liu, Keyuan; Hao, Xiaoyan; Li, Yang; Luo, Guobin; Zhang, Yonggen; Xin, Hangshu

    2017-06-26

    This study aims to identify the relationship between odd- and branched-chain fatty acids (OBCFAs) and microbial nucleic acid base in the rumen, and to establish a model to accurately predict microbial protein flow by using OBCFA. To develop the regression equations, data on the rumen contents of individual cows was obtained from 2 feeding experiments. In the first experiment, 3 rumen-fistulated dry dairy cows arranged in a 3×3 Latin square were fed diets of differing forage to concentration ratios (F:C). The second experiment consisted of 9 lactating Holstein dairy cows of similar body weights at the same fetal time. For each milk stage, 3 cows with similar milk production were selected. The rumen contents were sampled at 4 time points of every two hours after morning feeding 6 h, and then to analyse the concentrations of OBCFA and microbial nucleic acid bases in the rumen samples. The ruminal bacteria nucleic acid bases were significantly influenced by feeding diets of differing forge to concentration ratios and milk stages of dairy (P < 0.05). The concentrations of odd- and branched-chain fatty acids, especially odd-chain fatty acids and C15:0 isomers, strongly correlated with the microbial nucleic acid bases in the rumen (P < 0.05). The equations of ruminal microbial nucleic acid bases which established by ruminal OBCFAs contents showed a good a good predictive capacity, as indicated by reasonably low SEs and high R-squared values. This finding suggests that the rumen OBCFA composition could be used as an internal marker of rumen microbial matter.

  13. Evaluation of DNA extraction methods of rumen microbial populations.

    PubMed

    Villegas-Rivera, Gabriela; Vargas-Cabrera, Yevani; González-Silva, Napoleón; Aguilera-García, Florentino; Gutiérrez-Vázquez, Ernestina; Bravo-Patiño, Alejandro; Cajero-Juárez, Marcos; Baizabal-Aguirre, Víctor Manuel; Valdez-Alarcón, Juan José

    2013-02-01

    The dynamism of microbial populations in the rumen has been studied with molecular methods that analyze single nucleotide polymorphisms of ribosomal RNA gene fragments (rDNA). Therefore DNA of good quality is needed for this kind of analysis. In this work we report the evaluation of four DNA extraction protocols (mechanical lysis or chemical lysis with CTAB, ethylxanthogenate or DNAzol(®)) from ruminal fluid. The suitability of two of these protocols (mechanical lysis and DNAzol(®)) was tested on single-strand conformation polymorphism (SSCP) of rDNA of rumen microbial populations. DNAzol(®) was a simple method that rendered good integrity, yield and purity. With this method, subtle changes in protozoan populations were detected in young bulls fed with slightly different formulations of a supplement of multinutritional blocks of molasses and urea. Sequences related to Eudiplodinium maggi and a non-cultured Entodiniomorphid similar to Entodinium caudatum, were related to major fluctuating populations in an SSCP assay.

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

    PubMed

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

    2012-09-01

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

  15. Board-invited review: Rumen microbiology: Leading the way in microbial ecology

    USDA-ARS?s Scientific Manuscript database

    Robert Hungate, considered the father of rumen microbiology, was the first to initiate a systematic exploration of the microbial ecosystem of the rumen, but he was not alone. The techniques he developed to isolate and identify cellulose-digesting bacteria from the rumen have had a major impact not ...

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

    PubMed

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

    2008-11-01

    We evaluated the effect of increasing amounts of rumen-degradable intake protein (DIP) on urea kinetics in steers consuming prairie hay. Ruminally and duodenally fistulated steers (278 kg of BW) were used in a 4 x 4 Latin square and provided ad libitum access to low-quality prairie hay (4.9% CP). The DIP was provided as casein dosed ruminally once daily in amounts of 0, 59, 118, and 177 mg of N/kg of BW daily. Periods were 13 d long, with 7 d for adaptation and 6 d for collection. Steers were in metabolism crates for total collection of urine and feces. Jugular infusion of (15)N(15)N-urea, followed by determination of urinary enrichment of (15)N(15)N-urea and (14)N(15)N-urea was used to determine urea kinetics. Forage and N intake increased (linear, P < 0.001) with increasing DIP. Retention of N was negative (-2.7 g/d) for steers receiving no DIP and increased linearly (P < 0.001; 11.7, 23.0, and 35.2 g/d for 59, 118, and 177 mg of N/kg of BW daily) with DIP. Urea synthesis was 19.9, 24.8, 42.9, and 50.9 g of urea-N/d for 0, 59, 118, and 177 mg of N/kg of BW daily (linear, P = 0.004). Entry of urea into the gut was 98.9, 98.8, 98.6, and 95.9% of production for 0, 59, 118, and 177 mg of N/kg of BW daily, respectively (quadratic, P = 0.003). The amount of urea-N entering the gastrointestinal tract was greatest for 177 mg of N/kg of BW daily (48.6 g of urea-N/d) and decreased (linear, P = 0.005) to 42.4, 24.5, and 19.8 g of urea-N/d for 118, 59, and 0 mg of N/kg of BW daily. Microbial incorporation of recycled urea-N increased linearly (P = 0.02) from 12.3 g of N/d for 0 mg of N/kg of BW daily to 28.9 g of N/d for 177 mg of N/kg of BW daily. Provision of DIP produced the desired and previously observed increase in forage intake while also increasing N retention. The large percentage of urea synthesis that was recycled to the gut (95.9% even when steers received the greatest amount of DIP) points to the remarkable ability of cattle to conserve N when fed a low-protein

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

    PubMed Central

    Zhang, Litai; Huang, Xiaofeng; Xue, Bai; Peng, Quanhui; Wang, Zhisheng; Yan, Tianhai; Wang, Lizhi

    2015-01-01

    Vaccination through recombinant proteins against rumen methanogenesis provides a mitigation approach to reduce enteric methane (CH4) emissions in ruminants. The objective of present study was to evaluate the in vivo efficacy of a new vaccine candidate protein (EhaF) on methanogenesis and microbial population in the rumen of goats. We amplified the gene mru 1407 encoding protein EhaF using fresh rumen fluid samples of mature goats and successfully expressed recombinant protein (EhaF) in Escherichia coli Rosetta. This product was evaluated using 12 mature goats with half for control and other half injected with 400ug/goat the purified recombinant protein in day 1 and two subsequent booster immunizations in day 35 and 49. All measurements were undertaken from 63 to 68 days after the initial vaccination, with CH4 emissions determined using respiration calorimeter chambers. The results showed that the vaccination caused intensive immune responses in serum and saliva, although it had no significant effect on total enteric CH4 emissions and methanogen population in the rumen, when compared with the control goats. However, the vaccination altered the composition of rumen bacteria, especially the abundance of main phylum Firmicutes and genus Prevotella. The results indicate that protein EhaF might not be an effective vaccine to reduce enteric CH4 emissions but our vaccine have potential to influence the rumen ecosystem of goats. PMID:26445479

  18. Effects of Flavonoids on Rumen Fermentation Activity, Methane Production, and Microbial Population

    PubMed Central

    Abdullah, Norhani; Oskoueian, Armin

    2013-01-01

    This research was carried out to evaluate the effects of flavone, myricetin, naringin, catechin, rutin, quercetin, and kaempferol at the concentration of 4.5% of the substrate (dry matter basis) on the rumen microbial activity in vitro. Mixture of guinea grass and concentrate (60 : 40) was used as the substrate. The results showed that all the flavonoids except naringin and quercetin significantly (P < 0.05) decreased the dry matter degradability. The gas production significantly (P < 0.05) decreased by flavone, myricetin, and kaempferol, whereas naringin, rutin, and quercetin significantly (P < 0.05) increased the gas production. The flavonoids suppressed methane production significantly (P < 0.05). The total VFA concentration significantly (P < 0.05) decreased in the presence of flavone, myricetin, and kaempferol. All flavonoids except naringin and quercetin significantly (P < 0.05) reduced the carboxymethyl cellulase, filter paperase, xylanase, and β-glucosidase activities, purine content, and the efficiency of microbial protein synthesis. Flavone, myricetin, catechin, rutin, and kaempferol significantly (P < 0.05) reduced the population of rumen microbes. Total populations of protozoa and methanogens were significantly (P < 0.05) suppressed by naringin and quercetin. The results of this research demonstrated that naringin and quercetin at the concentration of 4.5% of the substrate (dry matter basis) were potential metabolites to suppress methane production without any negative effects on rumen microbial fermentation. PMID:24175289

  19. Abundance and Genetic Diversity of Microbial Polygalacturonase and Pectate Lyase in the Sheep Rumen Ecosystem

    PubMed Central

    Wang, Yaru; Luo, Huiying; Huang, Huoqing; Shi, Pengjun; Bai, Yingguo; Yang, Peilong; Yao, Bin

    2012-01-01

    Background Efficient degradation of pectin in the rumen is necessary for plant-based feed utilization. The objective of this study was to characterize the diversity, abundance, and functions of pectinases from microorganisms in the sheep rumen. Methodology/Principal Findings A total of 103 unique fragments of polygalacturonase (PF00295) and pectate lyase (PF00544 and PF09492) genes were retrieved from microbial DNA in the rumen of a Small Tail Han sheep, and 66% of the sequences of these fragments had low identities (<65%) with known sequences. Phylogenetic tree building separated the PF00295, PF00544, and PF09492 sequences into five, three, and three clades, respectively. Cellulolytic and noncellulolytic Butyrivibrio, Prevotella, and Fibrobacter species were the major sources of the pectinases. The two most abundant pectate lyase genes were cloned, and their protein products, expressed in Escherichia coli, were characterized. Both enzymes probably act extracellularly as their nucleotide sequences contained signal sequences, and they had optimal activities at the ruminal physiological temperature and complementary pH-dependent activity profiles. Conclusion/Significance This study reveals the specificity, diversity, and abundance of pectinases in the rumen ecosystem and provides two additional ruminal pectinases for potential industrial use under physiological conditions. PMID:22815874

  20. Influence of substrate and microbial interaction on efficiency of rumen microbial growth.

    PubMed

    Demeyer, D; Van Nevel, C

    1986-01-01

    Microbial N produced in the rumen and flowing to the duodenum (Ni) is related to the total amount of OM fermented or apparently digested in the rumen (OMf). This relationship, best expressed as microbial N yield (gNi/kgOMf), is affected mainly by the physical and chemical properties of feed carbohydrates and the amounts ingested. These factors influence yields at three levels of increasing complexity: Bacterial fermentation within one compartment following the continuous culture model. Fermentation pattern as such does not seem to affect yields. High fermentation rates are associated with lactate production, low methane production and transient polysaccharide synthesis. These effects induce acidification and lower yields, partly compensated by faster growth. Protozoal action, determined by the presence of sequestration spaces provided mainly by roughage diets. The presence of protozoa depresses microbial N yield but allows more complete fibre digestion. Compartmentation and differential passage. With roughage diets, optimal microbial N yield seems to require well developed microbial compartmentation, involving a large proportion of microbes in a large-particle pool with a slow turnover, balanced by a small proportion in liquid, small-particle pools with a fast turnover. Such a situation is associated with long roughage feeding. It is hypothesized that microbial N yields in the rumen may vary between two extremes which are associated with the feeding of long roughage on the one hand or with concentrate (starch) feeding on the other.

  1. Studies on potential effects of fumaric acid on rumen microbial fermentation, methane production and microbial community.

    PubMed

    Riede, Susanne; Boguhn, Jeannette; Breves, Gerhard

    2013-01-01

    The greenhouse gas methane (CH4) contributes substantially to global climate change. As a potential approach to decrease ruminal methanogenesis, the effects of different dosages of fumaric acid (FA) on ruminal microbial metabolism and on the microbial community (archaea, bacteria) were studied using a rumen simulation technique (RUSITEC). FA acts as alternative hydrogen acceptor diverting 2H from methanogenesis of archaea towards propionate formation of bacteria. Three identical trials were conducted with 12 fermentation vessels over a period of 14 days. In each trial, four fermentation vessels were assigned to one of the three treatment groups differing in FA dosage: low fumaric acid (LFA), high fumaric acid (HFA) and without FA (control). FA was continuously infused with the buffer. Grass silage and concentrate served as substrate. FA led to decreases in pH and to higher production rates of total short chain fatty acids (SCFA) mediated by increases in propionate for LFA of 1.69 mmol d(-1) and in propionate and acetate production for HFA of 4.49 and 1.10 mmol d(-1), respectively. Concentrations of NH3-N, microbial crude protein synthesis, their efficiency, degradation of crude nutrients and detergent fibre fraction were unchanged. Total gas and CH4 production were not affected by FA. Effects of FA on structure of microbial community by means of single strand conformation polymorphism (SSCP) analyses could not be detected. Given the observed increase in propionate production and the unaffected CH4 production it can be supposed that the availability of reduction equivalents like 2H was not limited by the addition of FA in this study. It has to be concluded from the present study that the application of FA is not an appropriate approach to decrease the ruminal CH4 production.

  2. Influence of supplemental protein source and feeding frequency on rumen fermentation and performance in dairy cows.

    PubMed

    Robinson, P H; McQueen, R E

    1994-05-01

    Multiparous Holstein cows in early lactation were fed a basal mixed ration of 47% (DM) alfalfa and timothy silage and 53% barley and corn concentrate twice daily for ad libitum intake at 1630 and 0600 h. Two supplemental protein sources that differed in their resistance to rumen proteolysis were fed at 9% of total DMI in either two meals per day at 1730 and 0700 h or five meals per day at 1730, 2130, 0200, 0700, and 1200 h. The study was a 4 x 4 Latin square design with six blocks of 4 cows in which one block of cows was fitted with rumen cannulas. Intakes of DM, OM, NDF, and CP were not influenced by treatments. However, cows supplemented with five meals a day tended to consume the mixed ration more rapidly after both the p.m. and a.m. feedings. Milk yield and its content of protein, fat, and lactose also were not influenced by treatments. Average rumen pH was higher, and propionate concentrations were lower, for cows supplemented with five meals, but diurnal patterns were not influenced. Propionate and rumen ammonia N concentrations were lower for cows supplemented with the more resistant protein source; however, rumen VFA, as well as soluble and peptide N concentrations, were not influenced by the type of supplemental protein. Results do not support benefits of synchronized rumen release of energy and N to overall cow production, but rather support previous research that soluble protein or peptide N, or both, may act as a pool to provide N for microbial growth at times of the day when ammonia N concentrations are very low.

  3. Effects of feed intake on composition of sheep rumen contents and their microbial population size.

    PubMed

    Rodríguez, C A; González, J; Alvir, M R; Redondo, R; Cajarville, C

    2003-01-01

    The present study was conducted to determine the effect of feed intake on the composition of the rumen contents of sheep and on their bacterial densities. Whole rumen contents were sampled after a period of continuous inter-rumen infusion of 15NH3 from four rumen-cannulated wethers successively fed on a hay-concentrate diet (2:1, w/w on a DM basis) at two rates of feed intake: 40 and 80 g DM/kg body weight0.75. Total weight and chemical composition of rumen contents, as well as the distribution by size and chemical composition of particles, were determined. The populations of bacteria associated with the liquid (liquid-associated bacteria, LAB) and solid (solid-associated bacteria, SAB) fractions of rumen digesta and the distribution of SAB according to feed particle size were also examined. The greater feed intake caused an increase in the mass of the rumen contents, while its chemical composition did not change, except for a higher content of organic matter (P=0.023). The distribution of feed particles by size was similar at both levels of intake. The concentrations of neutral- and acid-detergent fibre in feed particles decreased and those of total, dietary, and microbial N increased, both with a quadratic response (P=0.001), as particle size decreased. The proportion of LAB in the microbial biomass of rumen digesta reached only 8.0 %. This proportion and the density of LAB were unaffected by the level of feed intake, whereas an apparent reduction (10.4 %) occurred with the SAB biomass in whole rumen contents. A systematic, but not significant, reduction (mean value 11.9 %) in the level of microbial colonisation in the different particle fractions with the increase of feed intake was also observed.

  4. Extraction of PCR-quality plant and microbial DNA from total rumen contents.

    PubMed

    Sharma, Ranjana; John, S Jacob; Damgaard, M; McAllister, Tim A

    2003-01-01

    DNA from rumen digesta has several diagnostic applications such as studying microbial community dynamics, transgene/DNA stability, and population typing of various rumen bacteria. Several DNA extraction procedures are described in the literature for rumen digesta, which describe the removal of tannins, polysaccharides, and other PCR inhibitors. Some of these protocols are time-consuming and impractical when handling a large number of samples routinely. Here we describe a rapid method for the extraction of PCR-quality plant and microbial DNA from total rumen contents that is based on modifications in the cetyltrimethylammonium bromide procedure followed by cleanup using a Qiagen column. This procedure is highly reproducible and relatively short, once the initial grinding of the samples is performed, and it consistently yields PCR-quality DNA.

  5. Plant extracts affect in vitro rumen microbial fermentation.

    PubMed

    Busquet, M; Calsamiglia, S; Ferret, A; Kamel, C

    2006-02-01

    Different doses of 12 plant extracts and 6 secondary plant metabolites were incubated for 24 h in diluted ruminal fluid with a 50:50 forage:concentrate diet. Treatments were: control (no additive), plant extracts (anise oil, cade oil, capsicum oil, cinnamon oil, clove bud oil, dill oil, fenugreek, garlic oil, ginger oil, oregano oil, tea tree oil, and yucca), and secondary plant metabolites (anethol, benzyl salicylate, carvacrol, carvone, cinnamaldehyde, and eugenol). Each treatment was supplied at 3, 30, 300, and 3,000 mg/L of culture fluid. At 3,000 mg/L, most treatments decreased total volatile fatty acid concentration, but cade oil, capsicum oil, dill oil, fenugreek, ginger oil, and yucca had no effect. Different doses of anethol, anise oil, carvone, and tea tree oil decreased the proportion of acetate and propionate, which suggests that these compounds may not be nutritionally beneficial to dairy cattle. Garlic oil (300 and 3,000 mg/L) and benzyl salicylate (300 and 3,000 mg/L) reduced acetate and increased propionate and butyrate proportions, suggesting that methane production was inhibited. At 3,000 mg/L, capsicum oil, carvacrol, carvone, cinnamaldehyde, cinnamon oil, clove bud oil, eugenol, fenugreek, and oregano oil resulted in a 30 to 50% reduction in ammonia N concentration. Careful selection and combination of these extracts may allow the manipulation of rumen microbial fermentation.

  6. Parameters of rumen fermentation in a continuously fed sheep: evidence of a microbial rumination pool.

    PubMed

    Hungate, R E; Reichl, J; Prins, R

    1971-12-01

    The feed and feces of a continuously fed sheep were analyzed for carbon, hydrogen, and nitrogen, with oxygen as the remainder. The daily feed-feces weight difference was used as the reactant in an equation representing the rumen fermentation. The measured products were the daily production of volatile fatty acids (VFA), CH(4), CO(2), and ammonia. The carbon unaccounted for was assumed to be in the microbial cell material produced in the rumen and absorbed before reaching the feces. The ratio of C to H, O, and N in bacteria was used to represent the elemental composition of the microbes formed in the rumen fermentation, completing the following equation:C(20.03)H(36.99)O(17.406)N(1.345) + 5.65 H(2)O --> C(12)H(24)O(10.1) + 0.83 CH(4) VFA + 2.76 CO(2) + 0.50 NH(3) + C(4.44)H(8.88)O(2.35)N(0.785) microbial cells absorbed With C arbitrarily balanced and O balanced by appropriate addition of water, any error is reflected in the H. The H recovery was 98.5%. The turnover rate constant for rumen liquid equilibrating with polyethylene glycol (PEG) was 2.27 per day. Direct counts and volume measurements of the individual types of bacteria and protozoa in the rumen were used to calculate the total microbial cell volume in the rumen, not equilibrating with it. The dry matter in the rumen (582 g) and the nitrogen content (12.05) of the microbes in the rumen were estimated, the latter constituting 85% of the measured N in the rumen. Calculations for rumen dry matter and nitrogen turning over at the PEG rate introduce big discrepancies with other parameters; a rumination pool must be postulated. Its size and composition are estimated. Arguments are presented to support the view that dry matter and some of the microbes, chiefly the protozoa, do not leave the rumen at the PEG rate. One experiment with the same sheep fed twice daily showed significantly less production of microbial cells than did the continuous (each 2 hr) feeding. Analysis of the microbial cell yield suggests that

  7. Parameters of Rumen Fermentation in a Continuously Fed Sheep: Evidence of a Microbial Rumination Pool

    PubMed Central

    Hungate, R. E.; Reichl, J.; Prins, R.

    1971-01-01

    The feed and feces of a continuously fed sheep were analyzed for carbon, hydrogen, and nitrogen, with oxygen as the remainder. The daily feed-feces weight difference was used as the reactant in an equation representing the rumen fermentation. The measured products were the daily production of volatile fatty acids (VFA), CH4, CO2, and ammonia. The carbon unaccounted for was assumed to be in the microbial cell material produced in the rumen and absorbed before reaching the feces. The ratio of C to H, O, and N in bacteria was used to represent the elemental composition of the microbes formed in the rumen fermentation, completing the following equation:C20.03H36.99O17.406N1.345 + 5.65 H2O → C12H24O10.1 + 0.83 CH4 VFA + 2.76 CO2 + 0.50 NH3 + C4.44H8.88O2.35N0.785 microbial cells absorbed With C arbitrarily balanced and O balanced by appropriate addition of water, any error is reflected in the H. The H recovery was 98.5%. The turnover rate constant for rumen liquid equilibrating with polyethylene glycol (PEG) was 2.27 per day. Direct counts and volume measurements of the individual types of bacteria and protozoa in the rumen were used to calculate the total microbial cell volume in the rumen, not equilibrating with it. The dry matter in the rumen (582 g) and the nitrogen content (12.05) of the microbes in the rumen were estimated, the latter constituting 85% of the measured N in the rumen. Calculations for rumen dry matter and nitrogen turning over at the PEG rate introduce big discrepancies with other parameters; a rumination pool must be postulated. Its size and composition are estimated. Arguments are presented to support the view that dry matter and some of the microbes, chiefly the protozoa, do not leave the rumen at the PEG rate. One experiment with the same sheep fed twice daily showed significantly less production of microbial cells than did the continuous (each 2 hr) feeding. Analysis of the microbial cell yield suggests that, on the basis of 11 mg of cells

  8. Effect of field peas, chickpeas, and lentils on rumen fermentation, digestion, microbial protein synthesis, and feedlot performance in receiving diets for beef cattle.

    PubMed

    Gilbery, T C; Lardy, G P; Soto-Navarro, S A; Bauer, M L; Anderson, V L

    2007-11-01

    Two experiments were conducted to evaluate the use of pulse grains in receiving diets for cattle. In Exp. 1, 8 Holstein (615 +/- 97 kg of initial BW) and 8 Angus-crossbred steers (403 +/- 73 kg of initial BW) fitted with ruminal and duodenal cannulas were blocked by breed and used in a randomized complete block design to assess the effects of pulse grain inclusion in receiving diets on intake, ruminal fermentation, and site of digestion. Experiment 2 was a 39-d feedlot receiving trial in which 176 mixed-breed steers (254 +/- 19 kg of initial BW) were used in a randomized complete block design to determine the effects of pulse grains on DMI, ADG, and G:F in newly received feedlot cattle. In both studies, pulse grains (field peas, lentils, or chickpea) replaced corn and canola meal as the grain component in diets fed as a total mixed ration. Treatments included 1) corn and canola meal (control); 2) field pea; 3) lentil; and 4) chickpea. Preplanned orthogonal contrasts were conducted between control vs. chickpea, control vs. field pea, and control vs. lentil. In Exp. 1, there were no differences among treatments for DMI (11.63 kg/d, 2.32% of BW daily, P = 0.63) or OM intake (P = 0.63). No treatment effects for apparent ruminal (P = 0.10) and total tract OM digestibilities (P = 0.40) were detected when pulse grains replaced corn and canola meal. Crude protein intake (P = 0.78), microbial CP flow (P = 0.46), total tract CP digestibility (P = 0.45), and microbial efficiency (P = 0.18) were also not influenced by treatment. Total-tract ADF (P = 0.004) and NDF (P = 0.04) digestibilities were greater with field pea vs. control. Total VFA concentrations were lower for field pea (P = 0.009) and lentil (P < 0.001) compared with control. Chickpea, field pea, and lentil had lower (P < or = 0.03) acetate molar proportion than control. Ruminal pH (P = 0.18) and NH3 (P = 0.14) were not different among treatments. In Exp. 2, calves fed chickpea, field pea, and lentil had greater

  9. Methane Inhibition Alters the Microbial Community, Hydrogen Flow, and Fermentation Response in the Rumen of Cattle

    PubMed Central

    Martinez-Fernandez, Gonzalo; Denman, Stuart E.; Yang, Chunlei; Cheung, Jane; Mitsumori, Makoto; McSweeney, Christopher S.

    2016-01-01

    Management of metabolic hydrogen ([H]) in the rumen has been identified as an important consideration when reducing ruminant CH4 emissions. However, little is known about hydrogen flux and microbial rumen population responses to CH4 inhibition when animals are fed with slowly degradable diets. The effects of the anti-methanogenic compound, chloroform, on rumen fermentation, microbial ecology, and H2/CH4 production were investigated in vivo. Eight rumen fistulated Brahman steers were fed a roughage hay diet (Rhode grass hay) or roughage hay:concentrate diet (60:40) with increasing levels (low, mid, and high) of chloroform in a cyclodextrin matrix. The increasing levels of chloroform resulted in an increase in H2 expelled as CH4 production decreased with no effect on dry matter intakes. The amount of expelled H2 per mole of decreased methane, was lower for the hay diet suggesting a more efficient redirection of hydrogen into other microbial products compared with hay:concentrate diet. A shift in rumen fermentation toward propionate and branched-chain fatty acids was observed for both diets. Animals fed with the hay:concentrate diet had both higher formate concentration and H2 expelled than those fed only roughage hay. Metabolomic analyses revealed an increase in the concentration of amino acids, organic, and nucleic acids in the fluid phase for both diets when methanogenesis was inhibited. These changes in the rumen metabolism were accompanied by a shift in the microbiota with an increase in Bacteroidetes:Firmicutes ratio and a decrease in Archaea and Synergistetes for both diets. Within the Bacteroidetes family, some OTUs assigned to Prevotella were promoted under chloroform treatment. These bacteria may be partly responsible for the increase in amino acids and propionate in the rumen. No significant changes were observed for abundance of fibrolytic bacteria, protozoa, and fungi, which suggests that fiber degradation was not impaired. The observed 30% decrease in

  10. Sensory acceptability of raw and extruded bovine rumen protein in processed meat products.

    PubMed

    Conti-Silva, Ana Carolina; Silva, Maria Elisabeth Machado Pinto e; Arêas, José Alfredo Gomes

    2011-08-01

    The use of bovine rumen protein (raw and extruded) as a replacement for extruded soy protein concentrate in three meat products (pork sausage, chicken hamburger, and kibbe) was investigated. Similarity between rumen and soy protein meat products was assessed using triangle tests and sensory acceptability evaluated by consumer panelists using a nine-point hedonic scale. The addition of raw rumen protein was detected in all meat product types tested, while extruded rumen protein was only detected in kibbe. The addition of raw rumen protein decreased the acceptability of pork sausage aroma and flavor, but improved kibbe appearance, texture and overall acceptability. The addition of extruded rumen protein reduced the acceptability of chicken hamburger texture, but improved pork sausage flavor. Replacement of soy protein by bovine rumen protein is feasible based upon sensory results, but depended upon its form and the type of meat product to which it was added. Copyright © 2011 Elsevier Ltd. All rights reserved.

  11. Rumen transfaunation.

    PubMed

    DePeters, E J; George, L W

    2014-12-01

    The aim of this invited mini-review is to summarize the rumen transfaunation literature. Rumen transfaunation using the cud from a healthy donor animal to treat a sick recipient animal was practiced long before our understanding of rumen microorganisms. Around the mid-1900 s, scientists began to explore the benefits of rumen transfaunation and the associated microbial populations. Rumen transfaunation has been used clinically to treat indigestion and to enhance the return of normal rumen function following surgical correction of a left-displaced abomasum. Rumen transfaunation was also used to introduce unique rumen microorganisms into animals that were exposed to toxic compounds in plants. Rumen liquor contains chemical constituents that likely contribute to the beneficial effects of re-establishing a normal reticulo-rumen anaerobic fermentation. Recommendations for collecting rumen fluid, storage and volumes transferred are discussed. Rumen transfaunation is a common practice to treat indigestion on dairy and livestock operations. The support of a healthy microbial community in the digestive tract is also used for humans. Fecal microbiota transplantation has been used to treat digestive disorders in humans. Rumen transfaunation, although not widely studied with respect to mode of action, is an effective, practical, and easy method to treat simple indigestion of ruminants.

  12. Effect of DNA extraction methods and sampling techniques on the apparent structure of cow and sheep rumen microbial communities.

    PubMed

    Henderson, Gemma; Cox, Faith; Kittelmann, Sandra; Miri, Vahideh Heidarian; Zethof, Michael; Noel, Samantha J; Waghorn, Garry C; Janssen, Peter H

    2013-01-01

    Molecular microbial ecology techniques are widely used to study the composition of the rumen microbiota and to increase understanding of the roles they play. Therefore, sampling and DNA extraction methods that result in adequate yields of microbial DNA that also accurately represents the microbial community are crucial. Fifteen different methods were used to extract DNA from cow and sheep rumen samples. The DNA yield and quality, and its suitability for downstream PCR amplifications varied considerably, depending on the DNA extraction method used. DNA extracts from nine extraction methods that passed these first quality criteria were evaluated further by quantitative PCR enumeration of microbial marker loci. Absolute microbial numbers, determined on the same rumen samples, differed by more than 100-fold, depending on the DNA extraction method used. The apparent compositions of the archaeal, bacterial, ciliate protozoal, and fungal communities in identical rumen samples were assessed using 454 Titanium pyrosequencing. Significant differences in microbial community composition were observed between extraction methods, for example in the relative abundances of members of the phyla Bacteroidetes and Firmicutes. Microbial communities in parallel samples collected from cows by oral stomach-tubing or through a rumen fistula, and in liquid and solid rumen digesta fractions, were compared using one of the DNA extraction methods. Community representations were generally similar, regardless of the rumen sampling technique used, but significant differences in the abundances of some microbial taxa such as the Clostridiales and the Methanobrevibacter ruminantium clade were observed. The apparent microbial community composition differed between rumen sample fractions, and Prevotellaceae were most abundant in the liquid fraction. DNA extraction methods that involved phenol-chloroform extraction and mechanical lysis steps tended to be more comparable. However, comparison of data

  13. Effect of DNA Extraction Methods and Sampling Techniques on the Apparent Structure of Cow and Sheep Rumen Microbial Communities

    PubMed Central

    Henderson, Gemma; Cox, Faith; Kittelmann, Sandra; Miri, Vahideh Heidarian; Zethof, Michael; Noel, Samantha J.; Waghorn, Garry C.; Janssen, Peter H.

    2013-01-01

    Molecular microbial ecology techniques are widely used to study the composition of the rumen microbiota and to increase understanding of the roles they play. Therefore, sampling and DNA extraction methods that result in adequate yields of microbial DNA that also accurately represents the microbial community are crucial. Fifteen different methods were used to extract DNA from cow and sheep rumen samples. The DNA yield and quality, and its suitability for downstream PCR amplifications varied considerably, depending on the DNA extraction method used. DNA extracts from nine extraction methods that passed these first quality criteria were evaluated further by quantitative PCR enumeration of microbial marker loci. Absolute microbial numbers, determined on the same rumen samples, differed by more than 100-fold, depending on the DNA extraction method used. The apparent compositions of the archaeal, bacterial, ciliate protozoal, and fungal communities in identical rumen samples were assessed using 454 Titanium pyrosequencing. Significant differences in microbial community composition were observed between extraction methods, for example in the relative abundances of members of the phyla Bacteroidetes and Firmicutes. Microbial communities in parallel samples collected from cows by oral stomach-tubing or through a rumen fistula, and in liquid and solid rumen digesta fractions, were compared using one of the DNA extraction methods. Community representations were generally similar, regardless of the rumen sampling technique used, but significant differences in the abundances of some microbial taxa such as the Clostridiales and the Methanobrevibacter ruminantium clade were observed. The apparent microbial community composition differed between rumen sample fractions, and Prevotellaceae were most abundant in the liquid fraction. DNA extraction methods that involved phenol-chloroform extraction and mechanical lysis steps tended to be more comparable. However, comparison of data

  14. Metagenomic insights into the rumen microbial fibrolytic enzymes in Indian crossbred cattle fed finger millet straw.

    PubMed

    Jose, V Lyju; Appoothy, Thulasi; More, Ravi P; Arun, A Sha

    2017-12-01

    The rumen is a unique natural habitat, exhibiting an unparalleled genetic resource of fibrolytic enzymes of microbial origin that degrade plant polysaccharides. The objectives of this study were to identify the principal plant cell wall-degrading enzymes and the taxonomic profile of rumen microbial communities that are associated with it. The cattle rumen microflora and the carbohydrate-active enzymes were functionally classified through a whole metagenomic sequencing approach. Analysis of the assembled sequences by the Carbohydrate-active enzyme analysis Toolkit identified the candidate genes encoding fibrolytic enzymes belonging to different classes of glycoside hydrolases(11,010 contigs), glycosyltransferases (6366 contigs), carbohydrate esterases (4945 contigs), carbohydrate-binding modules (1975 contigs), polysaccharide lyases (480 contigs), and auxiliary activities (115 contigs). Phylogenetic analysis of CAZyme encoding contigs revealed that a significant proportion of CAZymes were contributed by bacteria belonging to genera Prevotella, Bacteroides, Fibrobacter, Clostridium, and Ruminococcus. The results indicated that the cattle rumen microbiome and the CAZymes are highly complex, structurally similar but compositionally distinct from other ruminants. The unique characteristics of rumen microbiota and the enzymes produced by resident microbes provide opportunities to improve the feed conversion efficiency in ruminants and serve as a reservoir of industrially important enzymes for cellulosic biofuel production.

  15. The microbial composition and metabolic potential of the ovine rumen

    USDA-ARS?s Scientific Manuscript database

    The rumen is efficient at biotransforming nitroaromatic explosive compounds, such as TNT, RDX, and HMX, which have been used widely in US military munitions. These compounds are present in > 4,000 military items, from large bombs to very small igniters. However, their manufacturing processes have g...

  16. Rumen Microbial Population Dynamics during Adaptation to a High-Grain Diet ▿ †

    PubMed Central

    Fernando, S. C.; Purvis, H. T.; Najar, F. Z.; Sukharnikov, L. O.; Krehbiel, C. R.; Nagaraja, T. G.; Roe, B. A.; DeSilva, U.

    2010-01-01

    High-grain adaptation programs are widely used with feedlot cattle to balance enhanced growth performance against the risk of acidosis. This adaptation to a high-grain diet from a high-forage diet is known to change the rumen microbial population structure and help establish a stable microbial population within the rumen. Therefore, to evaluate bacterial population dynamics during adaptation to a high-grain diet, 4 ruminally cannulated beef steers were adapted to a high-grain diet using a step-up diet regimen containing grain and hay at ratios of 20:80, 40:60, 60:40, and 80:20. The rumen bacterial populations were evaluated at each stage of the step-up diet after 1 week of adaptation, before the steers were transitioned to the next stage of the diet, using terminal restriction fragment length polymorphism (T-RFLP) analysis, 16S rRNA gene libraries, and quantitative real-time PCR. The T-RFLP analysis displayed a shift in the rumen microbial population structure during the final two stages of the step-up diet. The 16S rRNA gene libraries demonstrated two distinct rumen microbial populations in hay-fed and high-grain-fed animals and detected only 24 common operational taxonomic units out of 398 and 315, respectively. The 16S rRNA gene libraries of hay-fed animals contained a significantly higher number of bacteria belonging to the phylum Fibrobacteres, whereas the 16S rRNA gene libraries of grain-fed animals contained a significantly higher number of bacteria belonging to the phylum Bacteroidetes. Real-time PCR analysis detected significant fold increases in the Megasphaera elsdenii, Streptococcus bovis, Selenomonas ruminantium, and Prevotella bryantii populations during adaptation to the high-concentrate (high-grain) diet, whereas the Butyrivibrio fibrisolvens and Fibrobacter succinogenes populations gradually decreased as the animals were adapted to the high-concentrate diet. This study evaluates the rumen microbial population using several molecular approaches and

  17. Dietary supplementation of Rosmarinus officinalis L. leaves in sheep affects the abundance of rumen methanogens and other microbial populations.

    PubMed

    Cobellis, Gabriella; Yu, Zhongtang; Forte, Claudio; Acuti, Gabriele; Trabalza-Marinucci, Massimo

    2016-01-01

    Rumen microbiome has a great influence on ruminant health and productivity. Different plant extracts have been tested for their ability to modulate the rumen microbiome to improve feed digestion and fermentation. Among the evaluated plant extracts, essential oils, tannins, and saponins appeared to have positive effects on rumen protein metabolism, volatile fatty acids production, and methane and ammonia production. The objective of this study was to evaluate the effect of rosemary (Rosmarinus officinalis L.) leaves and essential oils on rumen microbial populations. Four ruminally cannulated sheep were used in a 4×4 Latin square design fed (21 d/period): 1) a control diet composed of alfalfa hay and concentrate pellet (CTR), 2) CTR supplemented with 7 g/d/sheep of rosemary essential oil adsorbed on an inert support (EO), 3) CTR with 10 g/d/sheep of dried and ground rosemary leaves (RL), and 4) CTR with 10 g/d of dried and ground rosemary leaves pelleted into concentrate (RL pellet). Abundance of total bacteria, archaea, protozoa, and some select bacterial species or groups was quantified using qPCR, while the community of bacteria and archaea was profiled using denaturing gradient gel electrophoresis. No difference in abundance was noted for total bacteria, protozoa, or Ruminococcus flavefaciens between the control and the treatments, but the rosemary leaves, either in loose form or in pellet, decreased the abundance of archaea and the genus Prevotella (P < 0.001). The rosemary leaves in loose form also decreased (P < 0.001) the abundance of Ruminococcus albus and Clostridium aminophilum, while the EO increased (P < 0.001) the abundance of Fibrobacter succinogenes. The community of bacteria and archaea was not affected by any of the supplements. Being able to affect the abundance of several groups of rumen microbes that are known to be involved in degradation of protein and fiber and production of methane and ammonia, rosemary leaves may be used to

  18. Effects of low rumen-degradable protein or abomasal fructan infusion on diet digestibility and urinary nitrogen excretion in lactating dairy cows.

    PubMed

    Gressley, T F; Armentano, L E

    2007-03-01

    Post-ileal carbohydrate fermentation in dairy cows converts blood urea nitrogen (BUN) into fecal microbial protein. This should reduce urinary N, increase fecal N, and reduce manure NH3 volatilization. However, if intestinal BUN recycling competes with ruminal BUN recycling, hindgut fermentation may reduce NH3 for rumen microbial protein synthesis. Eight lactating Holstein cows were used in a replicated 4 x 4 Latin square design with 14-d periods. Treatments were arranged as a 2 x 2 factorial. Diets contained either adequate rumen-degradable protein (RDP; high RDP) or were 28% below predicted RDP requirements (low RDP). Cows received abomasal infusions of either 10 L/d of saline or 10 L/d of saline containing 1 kg/d of inulin. We hypothesized that reducing ruminal NH3, either by restricting RDP intake or by diverting BUN to feces with inulin, would reduce rumen microbial protein synthesis, as would be evidenced by significant main effects of treatments on rumen NH3, milk production, and urinary purine derivative excretion. Furthermore, we thought it likely that effects of inulin might be greater when rumen NH3 was already low, as would be indicated by significant interactions between inulin infusion and dietary RDP level on rumen NH3, milk production, and urinary purine derivative excretion. Rumen NH3 was reduced by the low-RDP diet, but urinary purine derivative excretion and milk production were unaffected. However, the low-RDP diet reduced apparent total tract digestibility of OM and starch and reduced in situ rumen NDF digestibility. Abomasal inulin reduced the BUN concentration but did not affect milk yield or rumen NH3, suggesting that RDP requirements are not affected by hindgut fermentation. Inulin shifted 23 g/d of N from urine to feces. However, based on fecal purine excretion, we estimated that only 8 g/d of the increased fecal N was due to increased fecal microbial output. Inulin reduced true digestibility of dietary protein or increased nonmicrobial as

  19. Efficient anaerobic transformation of raw wheat straw by a robust cow rumen-derived microbial consortium.

    PubMed

    Lazuka, Adèle; Auer, Lucas; Bozonnet, Sophie; Morgavi, Diego P; O'Donohue, Michael; Hernandez-Raquet, Guillermina

    2015-11-01

    A rumen-derived microbial consortium was enriched on raw wheat straw as sole carbon source in a sequential batch-reactor (SBR) process under strict mesophilic anaerobic conditions. After five cycles of enrichment the procedure enabled to select a stable and efficient lignocellulolytic microbial consortium, mainly constituted by members of Firmicutes and Bacteroidetes phyla. The enriched community, designed rumen-wheat straw-derived consortium (RWS) efficiently hydrolyzed lignocellulosic biomass, degrading 55.5% w/w of raw wheat straw over 15days at 35°C and accumulating carboxylates as main products. Cellulolytic and hemicellulolytic activities, mainly detected on the cell bound fraction, were produced in the earlier steps of degradation, their production being correlated with the maximal lignocellulose degradation rates. Overall, these results demonstrate the potential of RWS to convert unpretreated lignocellulosic substrates into useful chemicals.

  20. Microbial degradation of usnic acid in the reindeer rumen

    NASA Astrophysics Data System (ADS)

    Sundset, Monica A.; Barboza, Perry S.; Green, Thomas K.; Folkow, Lars P.; Blix, Arnoldus Schytte; Mathiesen, Svein D.

    2010-03-01

    Reindeer ( Rangifer tarandus) eat and utilize lichens as an important source of energy and nutrients in winter. Lichens synthesize and accumulate a wide variety of phenolic secondary compounds, such as usnic acid, as a defense against herbivores and to protect against damage by UV-light in solar radiation. We have examined where and to what extent these phenolic compounds are degraded in the digestive tract of the reindeer, with particular focus on usnic acid. Three male reindeer were given ad libitum access to a control diet containing no usnic acid for three weeks and then fed lichens ad libitum (primarily Cladonia stellaris) containing 9.1 mg/g DM usnic acid for 4 weeks. Usnic acid intake in reindeer on the lichen diet was 91-117 mg/kg BM/day. In spite of this, no trace of usnic acid or conjugates of usnic acid was found either in fresh rumen fluid, urine, or feces. This suggests that usnic acid is rapidly degraded by rumen microbes, and that it consequently is not absorbed by the animal. This apparent ability to detoxify lichen phenolic compounds may gain increased importance with future enhanced UV-B radiation expected to cause increased protective usnic acid/phenol production in lichens.

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

    PubMed

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

    2013-07-01

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

  2. Fatty acid profiles associated with microbial colonization of freshly ingested grass and rumen biohydrogenation.

    PubMed

    Kim, E J; Sanderson, R; Dhanoa, M S; Dewhurst, R J

    2005-09-01

    Two in situ studies were conducted to examine the use of odd-chain fatty acid profiles to study microbial colonization of freshly ingested herbage in the rumen as well as fatty acid biohydrogenation. In the first study, fresh perennial ryegrass was subjected to a range of sample preparation methods before incubation in the rumen for 2 or 7 h. In the second study, fresh perennial ryegrass was chopped into 1-cm lengths and incubated in polyester bags in the rumen for 2, 8, and 24 h. After removal of bags from the rumen, 4 different washing methods, ranging from manual squeezing to machine washing, were applied. Fatty acids were extracted from washed residues and determined, as methyl esters, by gas chromatography. The main odd-chain fatty acids (with the exception of anteiso C(15:0)) were not found in fresh grass and were useful markers of the effects of incubation time, sample preparation method, and washing method on microbial colonization/contamination. The concentration of these and other odd-chain fatty acids increased with incubation time in both studies. The results indicate rapid and continued microbial colonization of freshly ingested forages, although patterns of odd-chain fatty acids did not reveal any further information about the types of bacteria-colonizing herbage. Principal component, biplot analysis provided a useful overall description of the processes of microbial colonization and degradation of plant fatty acids on fresh herbage incubated in the rumen. Bolus formation during mastication and ingestion results in extensive damage to herbage; none of the techniques (cutting, crushing, and drying/grinding) investigated in this work was able to replicate the effects of bolus formation in the animal. The study provided further evidence of loss of unfermented feed particles through polyester bag pores, especially when feeds are dried and ground. Biohydrogenation of the polyunsaturated fatty acids of fresh herbage was used principally by solid

  3. Buccal Swabbing as a Noninvasive Method To Determine Bacterial, Archaeal, and Eukaryotic Microbial Community Structures in the Rumen

    PubMed Central

    Kirk, Michelle R.; Jonker, Arjan; McCulloch, Alan

    2015-01-01

    Analysis of rumen microbial community structure based on small-subunit rRNA marker genes in metagenomic DNA samples provides important insights into the dominant taxa present in the rumen and allows assessment of community differences between individuals or in response to treatments applied to ruminants. However, natural animal-to-animal variation in rumen microbial community composition can limit the power of a study considerably, especially when only subtle differences are expected between treatment groups. Thus, trials with large numbers of animals may be necessary to overcome this variation. Because ruminants pass large amounts of rumen material to their oral cavities when they chew their cud, oral samples may contain good representations of the rumen microbiota and be useful in lieu of rumen samples to study rumen microbial communities. We compared bacterial, archaeal, and eukaryotic community structures in DNAs extracted from buccal swabs to those in DNAs from samples collected directly from the rumen by use of a stomach tube for sheep on four different diets. After bioinformatic depletion of potential oral taxa from libraries of samples collected via buccal swabs, bacterial communities showed significant clustering by diet (R = 0.37; analysis of similarity [ANOSIM]) rather than by sampling method (R = 0.07). Archaeal, ciliate protozoal, and anaerobic fungal communities also showed significant clustering by diet rather than by sampling method, even without adjustment for potentially orally associated microorganisms. These findings indicate that buccal swabs may in future allow quick and noninvasive sampling for analysis of rumen microbial communities in large numbers of ruminants. PMID:26276109

  4. Effect of Soybean Meal and Soluble Starch on Biogenic Amine Production and Microbial Diversity Using In vitro Rumen Fermentation.

    PubMed

    Jeong, Chang-Dae; Mamuad, Lovelia L; Kim, Seon-Ho; Choi, Yeon Jae; Soriano, Alvin P; Cho, Kwang Keun; Jeon, Che-Ok; Lee, Sung Sil; Lee, Sang-Suk

    2015-01-01

    ), R8 (Selenomonas ruminantium gene) and R9 (Selenomonas ruminantium strain LongY6) were found in samples with higher proportions of SS. Different feed ratios affect rumen fermentation in terms of pH, NH3-N, CH4, BA, volatile fatty acid and other metabolite concentrations and microbial diversity. Balanced protein and carbohydrate ratios are needed for rumen fermentation.

  5. Microbial populations and fermentation profiles in rumen liquid and solids of Holstein cows respond differently to dietary barley processing.

    PubMed

    Metzler-Zebeli, B U; Khol-Parisini, A; Gruber, L; Zebeli, Q

    2015-12-01

    To evaluate the effects of treating barley grain with lactic acid (LA) and heat on postprandial dynamics of 19 microbial taxa and fermentation in the rumen of dairy cows. This study was designed as a double 3 × 3 Latin square with six rumen-cannulated cows and three diets either containing untreated control barley or barley treated with 1% LA and 1% LA and heat (LAH, 55°C). Microbial populations, pH and volatile fatty acids were assessed in rumen liquid and solids during the postprandial period. Propionate increased and butyrate decreased in rumen solids of cows fed LA and LAH treated barley compared to the control barley. The LA but not LAH treatment depressed Fibrobacter succinogenes in rumen liquid and solids, whereas the opposite effect was observed for Ruminococcus albus in both fractions and Ruminococcus flavefaciens in rumen solids. LA promoted Ruminobacter amylophilus with the effect being more pronounced with LAH. The Lactobacillus group and Megasphaera elsdenii increased in both fractions with LA but not with LAH. LA and LAH treatment of barley differently altered ruminal abundance of certain bacterial taxa and fungi and increased propionate fermentation in rumen solids, whereby LA and LAH effects were consistent and mostly independent of the rumen fraction and time after barley feeding. Results provided evidence that LA and LAH treatment of barley can enhance rumen propionate fermentation without adversely affecting rumen pH. As propionate is the major contributor to gluconeogenesis in ruminants, the present barley treatment may have practical application to enhance energy supply in dairy cows. © 2015 The Society for Applied Microbiology.

  6. Effects of harvest timing on estimates of rumen degradable protein from alfalfa forages.

    USDA-ARS?s Scientific Manuscript database

    Alfalfa (Medicago sativa L.) proteins ingested by dairy cows typically degrade at rapid rates, and a low percentage escapes the rumen intact. Our objective was to quantify rumen degradable intake protein (DIP) for 'Affinity ' alfalfa harvested over a range of ages (0, 5, 10, 15, and 20 d from Stage ...

  7. Effect of fumarate reducing bacteria on in vitro rumen fermentation, methane mitigation and microbial diversity.

    PubMed

    Mamuad, Lovelia; Kim, Seon Ho; Jeong, Chang Dae; Choi, Yeon Jae; Jeon, Che Ok; Lee, Sang-Suk

    2014-02-01

    The metabolic pathways involved in hydrogen (H(2)) production, utilization and the activity of methanogens are the important factors that should be considered in controlling methane (CH(4)) emissions by ruminants. H(2) as one of the major substrate for CH(4) production is therefore should be controlled. One of the strategies on reducing CH(4) is through the use of hydrogenotrophic microorganisms such as fumarate reducing bacteria. This study determined the effect of fumarate reducing bacteria, Mitsuokella jalaludinii, supplementation on in vitro rumen fermentation, CH(4) production, diversity and quantity. M. jalaludinii significantly reduced CH(4) at 48 and 72 h of incubation and significantly increased succinate at 24 h. Although not significantly different, propionate was found to be highest in treatment containing M. jalaludinii at 12 and 48 h of incubation. These results suggest that supplementation of fumarate reducing bacteria to ruminal fermentation reduces CH(4) production and quantity, increases succinate and changes the rumen microbial diversity.

  8. Electricity generation from cellulose by rumen microorganisms in microbial fuel cells.

    PubMed

    Rismani-Yazdi, Hamid; Christy, Ann D; Dehority, Burk A; Morrison, Mark; Yu, Zhongtang; Tuovinen, Olli H

    2007-08-15

    In microbial fuel cells (MFCs) bacteria generate electricity by mediating the oxidation of organic compounds and transferring the resulting electrons to an anode electrode. The objective of this study was to test the possibility of generating electricity with rumen microorganisms as biocatalysts and cellulose as the electron donor in two-compartment MFCs. The anode and cathode chambers were separated by a proton exchange membrane and graphite plates were used as electrodes. The medium in the anode chamber was inoculated with rumen microorganisms, and the catholyte in the cathode compartment was ferricyanide solution. Maximum power density reached 55 mW/m(2) (1.5 mA, 313 mV) with cellulose as the electron donor. Cellulose hydrolysis and electrode reduction were shown to support the production of current. The electrical current was sustained for over 2 months with periodic cellulose addition. Clarified rumen fluid and a soluble carbohydrate mixture, serving as the electron donors, could also sustain power output. Denaturing gradient gel electrophoresis (DGGE) of PCR amplified 16S rRNA genes revealed that the microbial communities differed when different substrates were used in the MFCs. The anode-attached and the suspended consortia were shown to be different within the same MFC. Cloning and sequencing analysis of 16S rRNA genes indicated that the most predominant bacteria in the anode-attached consortia were related to Clostridium spp., while Comamonas spp. abounded in the suspended consortia. The results demonstrated that electricity can be generated from cellulose by exploiting rumen microorganisms as biocatalysts, but both technical and biological optimization is needed to maximize power output. (c) 2007 Wiley Periodicals, Inc.

  9. Natural and artificial feeding management before weaning promote different rumen microbial colonization but not differences in gene expression levels at the rumen epithelium of newborn goats

    PubMed Central

    Abecia, Leticia; Jiménez, Elisabeth; Martínez-Fernandez, Gonzalo; Martín-García, A. Ignacio; Ramos-Morales, Eva; Pinloche, Eric; Denman, Stuart E.; Newbold, C. Jamie

    2017-01-01

    The aim of this work was to evaluate the effect of feeding management during the first month of life (natural with the mother, NAT, or artificial with milk replacer, ART) on the rumen microbial colonization and the host innate immune response. Thirty pregnant goats carrying two fetuses were used. At birth one kid was taken immediately away from the doe and fed milk replacer (ART) while the other remained with the mother (NAT). Kids from groups received colostrum during first 2 days of life. Groups of four kids (from ART and NAT experimental groups) were slaughtered at 1, 3, 7, 14, 21 and 28 days of life. On the sampling day, after slaughtering, the rumen content was sampled and epithelial rumen tissue was collected. Pyrosequencing analyses of the bacterial community structure on samples collected at 3, 7, 14 and 28 days showed that both systems promoted significantly different colonization patterns (P = 0.001). Diversity indices increased with age and were higher in NAT feeding system. Lower mRNA abundance was detected in TLR2, TLR8 and TLR10 in days 3 and 5 compared to the other days (7, 14, 21 and 28). Only TLR5 showed a significantly different level of expression according to the feeding system, presenting higher mRNA abundances in ART kids. PGLYRP1 showed significantly higher abundance levels in days 3, 5 and 7, and then experienced a decline independently of the feeding system. These observations confirmed a highly diverse microbial colonisation from the first day of life in the undeveloped rumen, and show that the colonization pattern substantially differs between pre-ruminants reared under natural or artificial milk feeding systems. However, the rumen epithelial immune development does not differentially respond to distinct microbial colonization patterns. PMID:28813529

  10. In depth analysis of rumen microbial and carbohydrate-active enzymes profile in Indian crossbred cattle.

    PubMed

    Jose, V Lyju; More, Ravi P; Appoothy, Thulasi; Arun, A Sha

    2017-04-01

    Rumen houses a plethora of symbiotic microorganisms empowering the host to hydrolyze plant lignocellulose. In this study, NGS based metagenomic approach coupled with bioinformatic analysis was employed to gain an insight into the deconstruction of lignocellulose by carbohydrate-active enzymes (CAZymes) in Indian crossbred Holstein-Friesian cattle. Cattle rumen metagenomic DNA was sequenced using Illumina-MiSeq and 1.9 gigabases of data generated with an average read length of 871 bp. Analysis of the assembled sequences by Pfam-based Carbohydrate-active enzyme Analysis Toolkit identified 17,164 putative protein-encoding CAZymes belonging to different families of glycoside hydrolases (7574), glycosyltransferases (5185), carbohydrate-binding modules (2418), carbohydrate esterases (1516), auxiliary activities (434) and polysaccharide lyases (37). Phylogenetic analysis of putative CAZymes revealed that a significant proportion of CAZymes were contributed by bacteria belonging to the phylum Bacteroidetes (40%), Firmicutes (30%) and Proteobacteria (10%). The comparative analysis of HF cross rumen metagenome with other herbivore metagenomes indicated that Indian crossbred cattle rumen is endowed with a battery of CAZymes that may play a central role in lignocellulose deconstruction. The extensive catalog of enzymes reported in our study that hydrolyzes plant lignocellulose biomass, can be further explored for the better feed utilization in ruminants and also for different industrial applications.

  11. Comparison of rumen microbial inhibition resulting from various essential oils isolated from relatively unpalatable plant species.

    PubMed

    Oh, H K; Jones, M B; Longhurst, W M

    1968-01-01

    Essential oils were isolated from eight plant species which were relatively unpalatable to sheep and deer. The inhibitory potency of these essential oils upon sheep and deer rumen microorganisms was compared, in terms of total gas and volatile fatty acid (VFA) production, by use of an anaerobic manometric technique. Inhibitory effects of oils from the eight plant species may be placed in four groups: (i) essential oils from vinegar weed (Trichostema lanceoletum) and California bay (Umbellularia californica) inhibited rumen microbial activity most; (ii) lesser inhibition was exhibited by rosemary (Rosmarinus officinalis) and California mugwort (Artemisia douglasiana) oils, followed by (iii) blue-gum eucalyptus (Eucalyptus globulus) and sagebrush (Artemisia tridentata) oils; and (iv) oils from Douglas fir (Psuedotsuga menziesii) and Jerusalem oak (chenopodium botrys) resulted in the least inhibition, when 0.3 ml of each oil was used. A highly significant correlation coefficient (r = 0.98(**)) between total gas and VFA production indicated the validity of either method to measure the activity of rumen microorganisms. Our results are discussed in relation to the hypothesis that the selectivity and voluntary consumption of ruminants are related to the characteristic odor and antibacterial action of essential oils isolated from relatively unpalatable plant species.

  12. Comparison of Rumen Microbial Inhibition Resulting from Various Essential Oils Isolated from Relatively Unpalatable Plant Species

    PubMed Central

    Oh, Hi Kon; Jones, M. B.; Longhurst, W. M.

    1968-01-01

    Essential oils were isolated from eight plant species which were relatively unpalatable to sheep and deer. The inhibitory potency of these essential oils upon sheep and deer rumen microorganisms was compared, in terms of total gas and volatile fatty acid (VFA) production, by use of an anaerobic manometric technique. Inhibitory effects of oils from the eight plant species may be placed in four groups: (i) essential oils from vinegar weed (Trichostema lanceoletum) and California bay (Umbellularia californica) inhibited rumen microbial activity most; (ii) lesser inhibition was exhibited by rosemary (Rosmarinus officinalis) and California mugwort (Artemisia douglasiana) oils, followed by (iii) blue-gum eucalyptus (Eucalyptus globulus) and sagebrush (Artemisia tridentata) oils; and (iv) oils from Douglas fir (Psuedotsuga menziesii) and Jerusalem oak (chenopodium botrys) resulted in the least inhibition, when 0.3 ml of each oil was used. A highly significant correlation coefficient (r = 0.98**) between total gas and VFA production indicated the validity of either method to measure the activity of rumen microorganisms. Our results are discussed in relation to the hypothesis that the selectivity and voluntary consumption of ruminants are related to the characteristic odor and antibacterial action of essential oils isolated from relatively unpalatable plant species. PMID:5636470

  13. Processing barley grain with lactic acid and tannic acid ameliorates rumen microbial fermentation and degradation of dietary fibre in vitro.

    PubMed

    Deckardt, Kathrin; Metzler-Zebeli, Barbara U; Zebeli, Qendrim

    2016-01-15

    Cereal grains are important ingredients of ruminant diets, but their rapid degradation seriously impairs rumen fermentation and the host's health. The goal of this study was to evaluate whether processing of barley grain with 1 or 5% lactic acid (LA) and 1 or 5% tannic acid (TA), without or with an additional heat treatment (1% LAH or 1% TAH), may affect microbial ruminal abundance, fermentation profile, and nutrient degradation in vitro. Processing with LA lowered (P < 0.05) the concentration of short-chain fatty acids (SCFAs), proportions of branched-chain SCFA, and the acetate-to-propionate ratio. Treatment with 1% TAH and 1% LAH lowered (P < 0.05) gene copy numbers of total protozoa, rumen lipopolysaccharide, and degradation of crude protein, and tended (P = 0.08) to lower the proportion of the genus Prevotella. Treatment of barley grain with 1% LA or 1% LAH stimulated Clostridium cluster XIV. Degradation of fibre was enhanced (P < 0.05) by all LA and TA treatments. Chemical and heat treatment of barley grain modulated the ruminal fermentation profile and enhanced fibre degradation; however, processing of grain with LA seems to be superior because this effect was not associated with a concomitant depression in ruminal degradation of organic matter. © 2015 Society of Chemical Industry.

  14. Changes in rumen microbial fermentation are due to a combined effect of type of diet and pH.

    PubMed

    Calsamiglia, S; Cardozo, P W; Ferret, A; Bach, A

    2008-03-01

    Low ruminal pH may occur when feeding high-concentrate diets. However, because the reduction in pH occurs at the same time as the amount of concentrate fed increases, the changes observed in rumen fermentation may be attributed to pH or the type of substrate being fermented. Our objective was to determine the contribution of pH and type of substrate being fermented to the changes observed in rumen fermentation after supplying a high-concentrate diet. Eight dual-flow, continuous culture fermenters (1,400 mL) were used in 4 periods to study the effect of pH and type of diet being fermented on rumen microbial fermentation. Temperature (39 degrees C), solid (5%/h), and liquid (10%/h) dilution rates, and feeding schedule were maintained constant. Treatments were the type of diet (FOR = 60% ryegrass and alfalfa hays and 40% concentrate; CON = 10% straw and 90% concentrate) and pH (4.9, 5.2, 5.5, 5.8, 6.1, 6.4, 6.7, and 7.0). Diets were formulated to have similar CP and ruminally undegradable protein levels. Data were analyzed as a mixed-effects model considering the linear, quadratic, and cubic effects of pH, the effects of diet, and their interactions. Semipartial correlations of each independent variable were calculated to estimate the contribution of each factor to the overall relationship. True digestion of OM and NDF were affected by pH, but not by type of diet. Total VFA were reduced by pH and were greater in CON than in FOR. Acetate and butyrate concentrations were reduced by pH but were not affected by diet. Propionate concentration increased as the pH decreased and was greater in CON than in FOR. Ammonia-N concentration decreased with decreasing pH and was lower in CON than in FOR. Microbial N flow was affected by pH, diet, and their interaction. Dietary N flow increased as pH decreased and was greater in CON than in FOR. The degradation of CP followed the opposite pattern, increasing as pH increased, and was less in CON than in FOR. The efficiency of microbial

  15. Oral Samples as Non-Invasive Proxies for Assessing the Composition of the Rumen Microbial Community.

    PubMed

    Tapio, Ilma; Shingfield, Kevin J; McKain, Nest; Bonin, Aurélie; Fischer, Daniel; Bayat, Ali R; Vilkki, Johanna; Taberlet, Pierre; Snelling, Timothy J; Wallace, R John

    2016-01-01

    Microbial community analysis was carried out on ruminal digesta obtained directly via rumen fistula and buccal fluid, regurgitated digesta (bolus) and faeces of dairy cattle to assess if non-invasive samples could be used as proxies for ruminal digesta. Samples were collected from five cows receiving grass silage based diets containing no additional lipid or four different lipid supplements in a 5 x 5 Latin square design. Extracted DNA was analysed by qPCR and by sequencing 16S and 18S rRNA genes or the fungal ITS1 amplicons. Faeces contained few protozoa, and bacterial, fungal and archaeal communities were substantially different to ruminal digesta. Buccal and bolus samples gave much more similar profiles to ruminal digesta, although fewer archaea were detected in buccal and bolus samples. Bolus samples overall were most similar to ruminal samples. The differences between both buccal and bolus samples and ruminal digesta were consistent across all treatments. It can be concluded that either proxy sample type could be used as a predictor of the rumen microbial community, thereby enabling more convenient large-scale animal sampling for phenotyping and possible use in future animal breeding programs aimed at selecting cattle with a lower environmental footprint.

  16. Oral Samples as Non-Invasive Proxies for Assessing the Composition of the Rumen Microbial Community

    PubMed Central

    Tapio, Ilma; Shingfield, Kevin J.; McKain, Nest; Bonin, Aurélie; Fischer, Daniel; Bayat, Ali R.; Vilkki, Johanna; Taberlet, Pierre; Snelling, Timothy J.; Wallace, R. John

    2016-01-01

    Microbial community analysis was carried out on ruminal digesta obtained directly via rumen fistula and buccal fluid, regurgitated digesta (bolus) and faeces of dairy cattle to assess if non-invasive samples could be used as proxies for ruminal digesta. Samples were collected from five cows receiving grass silage based diets containing no additional lipid or four different lipid supplements in a 5 x 5 Latin square design. Extracted DNA was analysed by qPCR and by sequencing 16S and 18S rRNA genes or the fungal ITS1 amplicons. Faeces contained few protozoa, and bacterial, fungal and archaeal communities were substantially different to ruminal digesta. Buccal and bolus samples gave much more similar profiles to ruminal digesta, although fewer archaea were detected in buccal and bolus samples. Bolus samples overall were most similar to ruminal samples. The differences between both buccal and bolus samples and ruminal digesta were consistent across all treatments. It can be concluded that either proxy sample type could be used as a predictor of the rumen microbial community, thereby enabling more convenient large-scale animal sampling for phenotyping and possible use in future animal breeding programs aimed at selecting cattle with a lower environmental footprint. PMID:26986467

  17. Metagenomic analysis of the rumen microbial community following inhibition of methane formation by a halogenated methane analog

    PubMed Central

    Denman, Stuart E.; Martinez Fernandez, Gonzalo; Shinkai, Takumi; Mitsumori, Makoto; McSweeney, Christopher S.

    2015-01-01

    Japanese goats fed a diet of 50% Timothy grass and 50% concentrate with increasing levels of the anti-methanogenic compound, bromochloromethane (BCM) were investigated with respect to the microbial population and functional shifts in the rumen. Microbial ecology methods identified species that exhibited positive and negative responses to the increasing levels of BCM. The methane-inhibited rumen appeared to adapt to the higher H2 levels by shifting fermentation to propionate which was mediated by an increase in the population of H2-consuming Prevotella and Selenomonas spp. Metagenomic analysis of propionate production pathways was dominated by genomic content from these species. Reductive acetogenic marker gene libraries and metagenomics analysis indicate that reductive acetogenic species do not play a major role in the BCM treated rumen. PMID:26528253

  18. The influence of an increased cobalt supply on ruminal parameters and microbial vitamin B12 synthesis in the rumen of dairy cows.

    PubMed

    Stemme, Kirsten; Lebzien, Peter; Flachowsky, Gerhard; Scholz, Henner

    2008-06-01

    The aim of the study was to examine the effects of an elevated dietary cobalt supply to dairy cows on rumen fermentation parameters and microbial vitamin B12 synthesis in the rumen. Five lactating dairy cows fitted with a ruminal and a duodenal cannula were subsequently fed either a ration containing only the native cobalt content (0.17 mg Co/ kg DM) or a ration supplemented with cobalt sulphate (0.29 mg Co/kg DM). The pH-value, the ammonia concentration as well as the concentration and the molar proportions of short chain fatty acids in the rumen were not significantly influenced by feeding the ration with the higher cobalt content. While there was no difference in microbial protein flow, the cobalamin flow at the duodenum was significantly elevated in supplemented animals (3.67 +/- 0.69 vs. 8.63 +/- 2.22 mg B12/d). The efficiency of cobalt utilisation for ruminal vitamin B12 synthesis was calculated to be 7.1 +/- 1.3% for the unsupplemented and 9.5 +/- 2.4% for the supplemented ration. Further investigation has to prove if there are any benefits for cows resulting from the elevated cobalamin synthesis measured, caused by feeding higher amounts of dietary cobalt.

  19. Comparative characterization of reticular and duodenal digesta and possibilities of estimating microbial outflow from the rumen based on reticular sampling in dairy cows.

    PubMed

    Hristov, A N

    2007-10-01

    The objective of this experiment was to investigate the possibility of estimating the outflow of nutrients and microbial protein from the rumen based on sampling reticular contents as an alternative to duodenal sampling. Microbial protein flow estimates were also compared with a third method based on sampling of ruminal contents. Reticular and duodenal digesta and ruminal contents were recovered from 4 cows used in a 4 x 4 Latin square design experiment, in which the ruminal effects of 4 exogenous enzyme preparations were studied. Large and small particulate and fluid markers were used to estimate digesta flow in a triple-marker model; 15N was used as a microbial marker. Reticular and duodenal digesta were segregated into small and large particles (SP and LP, respectively) and a fluid phase, and ruminal digesta was segregated into particulate and fluid phases. Compared with digesta recovered at the duodenum, reticular digesta had lower OM and greater NDF contents. The proportion of microbial N was notably greater in the fluid phase of reticular digesta. Ruminal outflow of DM and OM was greater (by 17 and 28%) and that of NDF was lower (by 14%) when estimated from duodenal compared with reticular samples. There was no difference in the estimated flow of starch and nonammonia and microbial N between the reticular and duodenal techniques. Microbial N flow estimated based on ruminal sampling was similar to those based on duodenal and reticular sampling. The ruminal method, however, grossly overestimated flow of DM, OM, and NDF. This study supports the concept that microbial protein outflow from the rumen can be measured based on sampling of ruminal or reticular digesta. The reticular sampling technique can also provide reliable estimates for ruminal digestibility of OM, N, and fiber fractions. These findings need to be confirmed in experiments with basal diets varying in structure and forage-to-concentrate ratios.

  20. Effects of an Abrupt Change in Ration from All Roughage to High Concentrate upon Rumen Microbial Numbers in Sheep1

    PubMed Central

    Grubb, Jean A.; Dehority, B. A.

    1975-01-01

    When three sheep were abruptly changed from a ration of 100% orchardgrass hay to 60% cracked corn-40% orchardgrass hay, fed at equal dry-matter intakes, significant increases in concentration were observed in the rumen microbial population. Bacterial numbers (colony counts) per gram of rumen contents did not appear to have stabilized within 21 days after the ration change; however, protozoan numbers per milliliter plateaued after 5 days. The concentration of cellulose-digesting bacteria varied considerably between animals and decreased in all animals with the change. Changes were observed in total and molar percentages of volatile fatty acids, which were typical for the two types of rations. Although the concentration of protozoa increased after the ration change, only minor differences were observed in their percent generic distribution. A significant decrease in rumen volume was measured in two of the three sheep with the change in ration; however, fluid turnover rates were not significantly affected. Rates of rumen dry-matter turnover were slower with the concentrate ration, although rumen dry-matter digestion was increased. Calculation of total bacterial numbers based on total rumen volume completely negated the effect of ration change in one animal, whereas total numbers in the other two animals were still significantly different between rations and very similar between animals. Adjustment of total protozoa numbers did not alter the trends seen previously with concentration values. PMID:1180549

  1. Effects of quebracho tannin extract on rumen fermentation and yield and composition of microbial mass in heifers.

    PubMed

    Dickhoefer, U; Ahnert, S; Susenbeth, A

    2016-04-01

    To determine the effects of incremental quebracho tannin extract (QTE) infusions on rumen fermentation and yield and composition of microbial mass, a study was conducted that consisted of 1 control period (Control I) without QTE dosing; 4 periods when all animals received 1, 2, 4, or 6% QTE of the diet (DM basis); and a second control period (Control II). Each period consisted of 9 d of adaptation, 6 d of urine collection, and 1 d of rumen sampling. Three rumen-fistulated heifers (495 kg BW [SE 14]) were offered 2.6 kg/d grass hay, 2.6 kg/d concentrates, and 60 g/d mineral-vitamin premix (as fed) in 2 equal meals. Half the daily QTE dosage was intraruminally administered at every meal. Urine and rumen fluids were analyzed for purine derivatives and short-chain fatty acids, respectively. Nitrogen and purine base (PB) concentrations were determined in liquid-associated microbes in rumen fluid (LAMF), liquid-associated microbes in the solid phase (LAMS), and particle-associated microbes (PAM). Increasing QTE dosages linearly increased propionate and butyrate proportions in rumen fluid ( = 0.37, = 0.004 and = 0.51, < 0.001, respectively). Instead, proportions of acetate ( = 0.75, < 0.001), isobutyrate ( = 0.66, < 0.001), and isovalerate ( = 0.49, < 0.001) and urinary purine derivatives excretions ( = 0.66, < 0.001) linearly decreased with increasing QTE infusions. Inconsistent differences were observed for LAMF, but concentrations of PB in LAMS ( ≤ 0.018) and of N in PAM ( < 0.001) were greater at 6% QTE than at Control I, II, and 1% QTE. Hence, low to moderate QTE dosages hamper rumen fermentation and microbial biomass yields. Alongside the nutritional consequences for the host, the methodological implications of these effects should be considered in studies evaluating the targeted use of tannins in ruminant feeding.

  2. Effects of a specific blend of essential oils on apparent nutrient digestion, rumen fermentation and rumen microbial populations in sheep fed a 50:50 alfalfa hay:concentrate diet

    PubMed Central

    Khateri, N.; Azizi, O.; Jahani-Azizabadi, H.

    2017-01-01

    Objective An experiment was conducted to investigate the effects of a specific mixture of essential oils (MEO), containing thyme, clove and cinnamon EO, on rumen microbial fermentation, nutrient apparent digestibility and blood metabolites in fistulated sheep. Methods Six sheep fitted with ruminal fistulas were used in a repeated measurement design with two 24-d periods to investigate the effect of adding MEO at 0 (control), 0.8, and 1.6 mL/d on apparent nutrient digestibility, rumen fermentation characteristics, rumen microbial population and blood chemical metabolites. Animals were fed with a 50:50 alfalfa hay:concentrate diet. Results Ruminal pH, total volatile fatty acids (VFA) concentration, molar proportion of individual VFA, acetate: propionate ratio and methane production were not affected with MEO. Relative to the control, Small peptides plus amino acid nitrogen and large peptides nitrogen concentration in rumen fluid were not affected with MEO supplementation; while, rumen fluid ammonia nitrogen concentration at 0 and 6 h after morning feeding in sheep fed with 1.6 mL/d of MEO was lower (p<0.05) compared to the control and 0.8 mL/d of MEO. At 0 h after morning feeding, ammonia nitrogen concentration was higher (p<0.05) in sheep fed 0.8 mL/d of MEO relative to 1.6 mL/d and control diet. Ruminal protozoa and hyper ammonia producing (HAP) bacteria counts were not affected by addition of MEO in the diet. Relative to the control, no changes were observed in the red and white blood cells, hemoglobin, hematocrit, glucose, beta-hydroxybutyric acid, cholesterol, total protein, albumin, blood urea nitrogen and aspartate aminotransferase and alanine aminotransferase concentration. Apparent total tract digestibility of dry matter, crude proten, organic matter, and neutral detergent fiber were not influenced by MEO supplementation. Conclusion The results of the present study suggested that supplementation of MEO may have limited effects on apparent nutrient

  3. Metabolism of soluble rapeseed meal (Brassica rapa L.) protein during incubations with buffered bovine rumen contents in vitro.

    PubMed

    Stefański, T; Ahvenjärvi, S; Huhtanen, P; Shingfield, K J

    2013-01-01

    Accurate quantitative information on the fate of dietary protein in the rumen is central to modern metabolizable protein systems developed to improve the efficiency of nitrogen utilization in ruminants. An in vitro method was developed to estimate the rate of soluble rapeseed meal (Brassica rapa L.) protein (SRMP) degradation. Unlabeled and (15)N-labeled solvent-extracted rapeseed meal were incubated alone or as an equal mixture (125 mg of N/L) with buffered rumen contents and a mixture of carbohydrates formulated to provide a constant source of fermentable energy during the course of all incubations. Incubations were made over 0.33, 0.67, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 6.0, 8.0, and 10.0 h. Enrichment of (14)N and (15)N isotopes in total N of ammonia (AN), soluble nonammonia (SNAN), and insoluble (ISN) fractions liberated during incubations with test proteins was determined. A model with 4 pools that accounted for both intracellular and extracellular N transformations was used to estimate the rate of SRMP degradation. Parameter values used in the model were adjusted based on the size of A(14)N, A(15)N, SNA(14)N, SNA(15)N, IS(14)N, and IS(15)N pools, measured at different time points during incubations with buffered rumen fluid. The mean rate of N degradation for SRMP was estimated at 0.126 (SD 0.0499) h(-1). No substantive difference in the rate of protein degradation or microbial protein synthesis was observed during incubations of labeled and unlabeled substrates with rumen fluid. In conclusion, combined use of data from incubations of unlabeled and (15)N-labeled rapeseed protein with buffered rumen inoculum provided sufficient information to allow for estimation of parameter values in a complex dynamic model of soluble protein degradation. Results indicate the potential of the technique to evaluate the degradability of SNAN of other dietary protein sources and implicate ruminal escape of soluble rapeseed protein as an important source of amino acids in ruminants

  4. Isolation and identification of cellulose-binding proteins from sheep rumen contents.

    PubMed

    Toyoda, Atsushi; Iio, Wataru; Mitsumori, Makoto; Minato, Hajime

    2009-03-01

    To extend our understanding of the mechanisms of plant cell wall degradation in the rumen, cellulose-binding proteins (CBPs) from the contents of a sheep rumen were directly isolated and identified using a metaproteomics approach. The rumen CBPs were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and some CBPs revealed endoglucanase activities toward carboxymethyl cellulose. Using mass spectrometry analyses, four CBPs were identified and annotated as known proteins from the predominant rumen cellulolytic bacterium Fibrobacter succinogenes: tetratricopeptide repeat domain protein, OmpA family protein, fibro-slime domain protein, and cellulose-binding endoglucanase F (EGF). Another CBP was identified as the cellulosomal glycosyl hydrolase family 6 exoglucanase, Cel6A, of Piromyces equi. F. succinogenes cells expressing EGF were found to be major members of the bacterial community on the surface or at the inner surface of hay stems by immunohistochemical analyses using anti-EGF antibody. The finding that four of the five CBPs isolated and identified from sheep rumen contents were from F. succinogenes indicates that F. succinogenes is significantly involved in cellulose degradation in the rumen.

  5. Effect of Phenotypic Residual Feed Intake and Dietary Forage Content on the Rumen Microbial Community of Beef Cattle

    PubMed Central

    Carberry, Ciara A.; Kenny, David A.; Han, Sukkyan; McCabe, Matthew S.

    2012-01-01

    Feed-efficient animals have lower production costs and reduced environmental impact. Given that rumen microbial fermentation plays a pivotal role in host nutrition, the premise that rumen microbiota may contribute to host feed efficiency is gaining momentum. Since diet is a major factor in determining rumen community structure and fermentation patterns, we investigated the effect of divergence in phenotypic residual feed intake (RFI) on ruminal community structure of beef cattle across two contrasting diets. PCR-denaturing gradient gel electrophoresis (DGGE) and quantitative PCR (qPCR) were performed to profile the rumen bacterial population and to quantify the ruminal populations of Entodinium spp., protozoa, Fibrobacter succinogenes, Ruminococcus flavefaciens, Ruminococcus albus, Prevotella brevis, the genus Prevotella, and fungi in 14 low (efficient)- and 14 high (inefficient)-RFI animals offered a low-energy, high-forage diet, followed by a high-energy, low-forage diet. Canonical correspondence and Spearman correlation analyses were used to investigate associations between physiological variables and rumen microbial structure and specific microbial populations, respectively. The effect of RFI on bacterial profiles was influenced by diet, with the association between RFI group and PCR-DGGE profiles stronger for the higher forage diet. qPCR showed that Prevotella abundance was higher (P < 0.0001) in inefficient animals. A higher (P < 0.0001) abundance of Entodinium and Prevotella spp. and a lower (P < 0.0001) abundance of Fibrobacter succinogenes were observed when animals were offered the low-forage diet. Thus, differences in the ruminal microflora may contribute to host feed efficiency, although this effect may also be modulated by the diet offered. PMID:22562991

  6. Effect of phenotypic residual feed intake and dietary forage content on the rumen microbial community of beef cattle.

    PubMed

    Carberry, Ciara A; Kenny, David A; Han, Sukkyan; McCabe, Matthew S; Waters, Sinead M

    2012-07-01

    Feed-efficient animals have lower production costs and reduced environmental impact. Given that rumen microbial fermentation plays a pivotal role in host nutrition, the premise that rumen microbiota may contribute to host feed efficiency is gaining momentum. Since diet is a major factor in determining rumen community structure and fermentation patterns, we investigated the effect of divergence in phenotypic residual feed intake (RFI) on ruminal community structure of beef cattle across two contrasting diets. PCR-denaturing gradient gel electrophoresis (DGGE) and quantitative PCR (qPCR) were performed to profile the rumen bacterial population and to quantify the ruminal populations of Entodinium spp., protozoa, Fibrobacter succinogenes, Ruminococcus flavefaciens, Ruminococcus albus, Prevotella brevis, the genus Prevotella, and fungi in 14 low (efficient)- and 14 high (inefficient)-RFI animals offered a low-energy, high-forage diet, followed by a high-energy, low-forage diet. Canonical correspondence and Spearman correlation analyses were used to investigate associations between physiological variables and rumen microbial structure and specific microbial populations, respectively. The effect of RFI on bacterial profiles was influenced by diet, with the association between RFI group and PCR-DGGE profiles stronger for the higher forage diet. qPCR showed that Prevotella abundance was higher (P < 0.0001) in inefficient animals. A higher (P < 0.0001) abundance of Entodinium and Prevotella spp. and a lower (P < 0.0001) abundance of Fibrobacter succinogenes were observed when animals were offered the low-forage diet. Thus, differences in the ruminal microflora may contribute to host feed efficiency, although this effect may also be modulated by the diet offered.

  7. Microbial population in the rumen of swamp buffalo (Bubalus bubalis) as influenced by coconut oil and mangosteen peel supplementation.

    PubMed

    Pilajun, R; Wanapat, M

    2013-06-01

    Four, rumen fistulated swamp buffalo bulls were used to study microbial populations in the rumen when supplemented with coconut oil and mangosteen peel. Animals were randomly assigned to a 4 × 4 Latin square design. Four treatments were un-supplemented (Control), supplementation with coconut oil at 50 g/kg (CO5), supplementation with mangosteen peel at 30 g/kg (MP3) and supplementation with CO5 and MP3 (COM), of total DM intake. Animals received concentrate at 10 g/kg of BW, and rice straw was given ad libitum. Abundance of total bacteria was increased by CO5 supplementation, whereas populations of protozoa and Fibrobacter succinogenes were reduced by CO5 and COM supplementation. Dietary supplementation did not affect methanogen, Ruminococcus flavefaciens or Ruminococcus albus abundances. Dietary treatments changed denaturing gradient gel electrophoresis (DGGE) band patterns of methanogens and protozoa when compared with the control group, especially when supplemented with MP3. Supplementation of COM resulted in the greatest difference in pattern of DGGE bands for total bacteria compared with the control. Coconut oil and mangosteen peel supplementation resulted in changing of rumen microbial abundances and communities; however, combination of them could be more benefit to improve rumen fermentation of swamp buffalo fed on rice straw. © 2012 Blackwell Verlag GmbH.

  8. Rumen microbial abundance and fermentation profile during severe subacute ruminal acidosis and its modulation by plant derived alkaloids in vitro.

    PubMed

    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.

  9. Dietary n-6:n-3 Fatty Acid Ratios Alter Rumen Fermentation Parameters and Microbial Populations in Goats.

    PubMed

    Ebrahimi, Mahdi; Rajion, Mohamed Ali; Adeyemi, Kazeem Dauda; Jafari, Saeid; Jahromi, Mohammad Faseleh; Oskoueian, Ehsan; Meng, Goh Yong; Ghaffari, Morteza Hosseini

    2017-02-01

    Revealing the ruminal fermentation patterns and microbial populations as affected by dietary n-6:n-3 PUFA ratio would be useful for further clarifying the role of the rumen in the lipid metabolism of ruminants. The objective of the present study was to investigate the effects of dietary n-6:n-3 PUFA ratios on fermentation characteristics, fatty acid (FA) profiles, and microbial populations in the rumen of goats. A total of twenty-one goats were randomly assigned to three dietary treatments with different n-6:n-3 PUFA ratios of 2.27:1 (low ratio, LR), 5.01:1 (medium ratio, MR), and 10.38:1 (high ratio, HR). After 100 days of feeding, all goats were slaughtered. Dietary n-6:n-3 PUFA ratios had no effect (P > 0.05) on rumen pH and NH3N concentration. Goats fed HR diet had lower (P < 0.05) propionate and total volatile fatty acids and higher (P < 0.05) butyrate compared with those fed the MR and LR diets. The proportion of C18:0 decreased (P < 0.05) as dietary n-6:n-3 PUFA ratios increased. The proportions of C18:1 trans-11, C18:2n-6, cis-9 trans-11 CLA, and C20:4n-6 were greater in the HR goats compared with the MR and LR goats. Lowering dietary n-6:n-3 PUFA ratios enhanced (P < 0.05) the proportion of C18:3n-3 and total n-3 PUFA in the rumen fluid of goats. The populations of R. albus and R. flavefaciens decreased (P < 0.05) as the n-6:n-3 PUFA ratios increased in diet. Diet had no effect (P > 0.05) on the ruminal populations of F. succinogenes, total bacteria, methanogens, total protozoa, Entiodinium, and Holotrich. The population of B. fibrisolvens was lower (P < 0.05) in the LR goats compared with the MR and HR goats. It was concluded that HR would increase the concentration of cis-9 trans-11 CLA and C18:1 trans-11 in the rumen. However, LR whould decrease the B. fibrisolvens population, which is involved in the BH process in the rumen. Further research is needed to evaluate the potential role and contribution of rumen microbiome in the metabolism of FA in the

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

    PubMed Central

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

    2015-01-01

    This study evaluated the in situ ruminal degradability, and subsequent small intestinal digestibility (SID) of dry matter, crude protein (CP), and amino acids (AA) of cottonseed meal (CSM), sunflower seed meal (SFSM) and distillers dried grains with solubles (DDGS) by using the modified three-step in vitro procedure. The ruminal degradability and subsequent SID of AA in rumen-undegradable protein (RUP-AA) varied among three protein supplements. The result show that the effective degradability of DM for SFSM, CSM, and DDGS was 60.8%, 56.4%, and 41.0% and their ruminal fermentable organic matter was 60.0%, 55.9%, and 39.9%, respectively. The ruminal degradable protein (RDP) content in CP for SFSM, CSM, and DDGS was 68.3%, 39.0%, and 32.9%, respectively, at the ruminal solid passage rate of 1.84%/h. The SFSM is a good source of RDP for rumen micro-organisms; however, the SID of RUP of SFSM was lower. The DDGS and CSM are good sources of RUP for lambs to digest in the small intestine to complement ruminal microbial AA of growing lambs. Individual RUP-AA from each protein source was selectively removed by the rumen micro-organisms, especially for Trp, Arg, His, and Lys (p<0.01). The SID of individual RUP-AA was different within specific RUP origin (p<0.01). Limiting amino acid was Leu for RUP of CSM and Lys for both RUP of SFSM and DDGS, respectively. Therefore, different protein supplements with specific limitations should be selected and combined carefully in growing lambs ration to optimize AA balance. PMID:25656208

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

    PubMed

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

    2015-04-01

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

  12. Rumen development process in goats as affected by supplemental feeding v. grazing: age-related anatomic development, functional achievement and microbial colonisation.

    PubMed

    Jiao, Jinzhen; Li, Xiaopeng; Beauchemin, Karen A; Tan, Zhiliang; Tang, Shaoxun; Zhou, Chuanshe

    2015-03-28

    The aim of the present study was to describe age-related changes in anatomic, functional and microbial variables during the rumen development process, as affected by the feeding system (supplemental feeding v. grazing), in goats. Goats were slaughtered at seven time points that were selected to reflect the non-rumination (0, 7 and 14 d), transition (28 and 42 d) and rumination (56 and 70 d) phases of rumen development. Total volatile fatty acid (TVFA) concentration (P= 0·002), liquid-associated bacterial and archaeal copy numbers (P< 0·01) were greater for supplemental feeding v. grazing, while rumen pH (P< 0·001), acetate molar proportion (P= 0·003) and solid-associated microbial copy numbers (P< 0·05) were less. Rumen papillae length (P= 0·097) and extracellular (P= 0·093) and total (P= 0·073) protease activity potentials in supplemented goats tended to be greater than those in grazing goats. Furthermore, from 0 to 70 d, irrespective of the feeding system, rumen weight, rumen wall thickness, rumen papillae length and area, TVFA concentration, xylanase, carboxymethylcellulase activity potentials, and microbial copy numbers increased (P< 0·01) with age, while the greatest amylase and protease activity potentials occurred at 28 d. Most anatomic and functional variables evolved progressively from 14 to 42 d, while microbial colonisation was fastest from birth to 28 d. These outcomes suggest that the supplemental feeding system is more effective in promoting rumen development than the grazing system; in addition, for both the feeding systems, microbial colonisation in the rumen is achieved at 1 month, functional achievement at 2 months, and anatomic development after 2 months.

  13. Effects of alfalfa flavonoids extract on the microbial flora of dairy cow rumen.

    PubMed

    Zhan, Jinshun; Liu, Mingmei; Wu, Caixia; Su, Xiaoshuang; Zhan, Kang; Zhao, Guo Qi

    2017-09-01

    The effect of flavonoids from alfalfa on the microbial flora was determined using molecular techniques of 16S ribosome deoxyribonucleic acid (rDNA) analysis. Four primiparous Holstein heifers fitted with ruminal cannulas were used in a 4×4 Latin square design and fed a total mixed ration to which alfalfa flavonoids extract (AFE) was added at the rates of 0 (A, control), 20 (B), 60 (C), or 100 (D) mg per kg of heifer BW. The number of operational taxonomic units in heifers given higher levels of flavonoid extract (C and D) was higher than for the two other treatments. The Shannon, Ace, and Chao indices for treatment C were significantly higher than for the other treatments (p<0.05). The number of phyla and genera increased linearly with increasing dietary supplementation of AFE (p<0.05). The principal co-ordinates analysis plot showed substantial differences in the microbial flora for the four treatments. The microbial flora in treatment A was similar to that in B, C, and D were similar by the weighted analysis. The richness of Tenericutes at the phylum level tended to increase with increasing AFE (p = 0.10). The proportion of Euryarchaeota at the phylum level increased linearly, whereas the proportion of Fusobacteria decreased linearly with increasing AFE supplementation (p = 0.04). The percentage of Mogibacterium, Pyramidobacter, and Asteroleplasma at the genus level decreased linearly with increasing AFE (p<0.05). The abundance of Spirochaeta, Succinivibrio, and Suttonella at the genus level tended to decrease linearly with increasing AFE (0.05microbial composition of the rumen; however its effect on nutrient digestibility remains to be determined.

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

    PubMed Central

    Henderson, Gemma; Cox, Faith; Ganesh, Siva; Jonker, Arjan; Young, Wayne; Abecia, Leticia; Angarita, Erika; Aravena, Paula; Nora Arenas, Graciela; Ariza, Claudia; Attwood, Graeme T.; Mauricio Avila, Jose; Avila-Stagno, Jorge; Bannink, André; Barahona, Rolando; Batistotti, Mariano; Bertelsen, Mads F.; Brown-Kav, Aya; Carvajal, Andres M.; Cersosimo, Laura; Vieira Chaves, Alexandre; Church, John; Clipson, Nicholas; Cobos-Peralta, Mario A.; Cookson, Adrian L.; Cravero, Silvio; Cristobal Carballo, Omar; Crosley, Katie; Cruz, Gustavo; Cerón Cucchi, María; de la Barra, Rodrigo; De Menezes, Alexandre B.; Detmann, Edenio; Dieho, Kasper; Dijkstra, Jan; dos Reis, William L. S.; Dugan, Mike E. R.; Hadi Ebrahimi, Seyed; Eythórsdóttir, Emma; Nde Fon, Fabian; Fraga, Martín; Franco, Francisco; Friedeman, Chris; Fukuma, Naoki; Gagić, Dragana; Gangnat, Isabelle; Javier Grilli, Diego; Guan, Le Luo; Heidarian Miri, Vahideh; Hernandez-Sanabria, Emma; Gomez, Alma Ximena Ibarra; Isah, Olubukola A.; Ishaq, Suzanne; Jami, Elie; Jelincic, Juan; Kantanen, Juha; Kelly, William J.; Kim, Seon-Ho; Klieve, Athol; Kobayashi, Yasuo; Koike, Satoshi; Kopecny, Jan; Nygaard Kristensen, Torsten; Julie Krizsan, Sophie; LaChance, Hannah; Lachman, Medora; Lamberson, William R.; Lambie, Suzanne; Lassen, Jan; Leahy, Sinead C.; Lee, Sang-Suk; Leiber, Florian; Lewis, Eva; Lin, Bo; Lira, Raúl; Lund, Peter; Macipe, Edgar; Mamuad, Lovelia L.; Cuquetto Mantovani, Hilário; Marcoppido, Gisela Ariana; Márquez, Cristian; Martin, Cécile; Martinez, Gonzalo; Eugenia Martinez, Maria; Lucía Mayorga, Olga; McAllister, Tim A.; McSweeney, Chris; Mestre, Lorena; Minnee, Elena; Mitsumori, Makoto; Mizrahi, Itzhak; Molina, Isabel; Muenger, Andreas; Munoz, Camila; Murovec, Bostjan; Newbold, John; Nsereko, Victor; O’Donovan, Michael; Okunade, Sunday; O’Neill, Brendan; Ospina, Sonia; Ouwerkerk, Diane; Parra, Diana; Pereira, Luiz Gustavo Ribeiro; Pinares-Patino, Cesar; Pope, Phil B.; Poulsen, Morten; Rodehutscord, Markus; Rodriguez, Tatiana; Saito, Kunihiko; Sales, Francisco; Sauer, Catherine; Shingfield, Kevin; Shoji, Noriaki; Simunek, Jiri; Stojanović-Radić, Zorica; Stres, Blaz; Sun, Xuezhao; Swartz, Jeffery; Liang Tan, Zhi; Tapio, Ilma; Taxis, Tasia M.; Tomkins, Nigel; Ungerfeld, Emilio; Valizadeh, Reza; van Adrichem, Peter; Van Hamme, Jonathan; Van Hoven, Woulter; Waghorn, Garry; John Wallace, R.; Wang, Min; Waters, Sinéad M.; Keogh, Kate; Witzig, Maren; Wright, Andre-Denis G.; Yamano, Hidehisa; Yan, Tianhai; Yanez-Ruiz, David R.; Yeoman, Carl J.; Zambrano, Ricardo; Zeitz, Johanna; Zhou, Mi; Wei Zhou, Hua; Xia Zou, Cai; Zunino, Pablo; Janssen, Peter H.

    2015-01-01

    Ruminant livestock are important sources of human food and global greenhouse gas emissions. Feed degradation and methane formation by ruminants rely on metabolic interactions between rumen microbes and affect ruminant productivity. Rumen and camelid foregut microbial community composition was determined in 742 samples from 32 animal species and 35 countries, to estimate if this was influenced by diet, host species, or geography. Similar bacteria and archaea dominated in nearly all samples, while protozoal communities were more variable. The dominant bacteria are poorly characterised, but the methanogenic archaea are better known and highly conserved across the world. This universality and limited diversity could make it possible to mitigate methane emissions by developing strategies that target the few dominant methanogens. Differences in microbial community compositions were predominantly attributable to diet, with the host being less influential. There were few strong co-occurrence patterns between microbes, suggesting that major metabolic interactions are non-selective rather than specific. PMID:26449758

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

    PubMed

    Henderson, Gemma; Cox, Faith; Ganesh, Siva; Jonker, Arjan; Young, Wayne; Janssen, Peter H

    2015-10-09

    Ruminant livestock are important sources of human food and global greenhouse gas emissions. Feed degradation and methane formation by ruminants rely on metabolic interactions between rumen microbes and affect ruminant productivity. Rumen and camelid foregut microbial community composition was determined in 742 samples from 32 animal species and 35 countries, to estimate if this was influenced by diet, host species, or geography. Similar bacteria and archaea dominated in nearly all samples, while protozoal communities were more variable. The dominant bacteria are poorly characterised, but the methanogenic archaea are better known and highly conserved across the world. This universality and limited diversity could make it possible to mitigate methane emissions by developing strategies that target the few dominant methanogens. Differences in microbial community compositions were predominantly attributable to diet, with the host being less influential. There were few strong co-occurrence patterns between microbes, suggesting that major metabolic interactions are non-selective rather than specific.

  16. Effects of different sources of physically effective fiber on rumen microbial populations.

    PubMed

    Shaw, C N; Kim, M; Eastridge, M L; Yu, Z

    2016-03-01

    Physically effective fiber is needed by dairy cattle to prevent ruminal acidosis. This study aimed to examine the effects of different sources of physically effective fiber on the populations of fibrolytic bacteria and methanogens. Five ruminally cannulated Holstein cows were each fed five diets differing in physically effective fiber sources over 15 weeks (21 days/period) in a Latin Square design: (1) 44.1% corn silage, (2) 34.0% corn silage plus 11.5% alfalfa hay, (3) 34.0% corn silage plus 5.1% wheat straw, (4) 36.1% corn silage plus 10.1% wheat straw, and (5) 34.0% corn silage plus 5.5% corn stover. The impact of the physically effective fiber sources on total bacteria and archaea were examined using denaturing gradient gel electrophoresis. Specific real-time PCR assays were used to quantify total bacteria, total archaea, the genus Butyrivibrio, Fibrobacter succinogenes, Ruminococcus albus, Ruminococcus flavefaciens and three uncultured rumen bacteria that were identified from adhering ruminal fractions in a previous study. No significant differences were observed among the different sources of physical effective fiber with respect to the microbial populations quantified. Any of the physically effective fiber sources may be fed to dairy cattle without negative impact on the ruminal microbial community.

  17. Metagenomic analysis of rumen microbial population in dairy heifers fed a high grain diet supplemented with dicarboxylic acids or polyphenols.

    PubMed

    De Nardi, Roberta; Marchesini, Giorgio; Li, Shucong; Khafipour, Ehsan; Plaizier, Kees J C; Gianesella, Matteo; Ricci, Rebecca; Andrighetto, Igino; Segato, Severino

    2016-02-19

    The aim of this study was to investigate the effects of two feed supplements on rumen bacterial communities of heifers fed a high grain diet. Six Holstein-Friesian heifers received one of the following dietary treatments according to a Latin square design: no supplement (control, C), 60 g/day of fumarate-malate (organic acid, O) and 100 g/day of polyphenol-essential oil (P). Rumen fluid was analyzed to assess the microbial population using Illumina sequencing and quantitative real time PCR. The P treatment had the highest number of observed species (P < 0.10), Chao1 index (P < 0.05), abundance based coverage estimated (ACE) (P < 0.05), and Fisher's alpha diversity (P < 0.10). The O treatment had intermediate values between C and P treatments with the exception of the Chao1 index. The PCoA with unweighted Unifrac distance showed a separation among dietary treatments (P = 0.09), above all between the C and P (P = 0.05). The O and P treatments showed a significant increase of the family Christenenellaceae and a decline of Prevotella brevis compared to C. Additionally, the P treatment enhanced the abundance of many taxa belonging to Bacteroidetes, Firmicutes and Tenericutes phyla due to a potential antimicrobial activity of flavonoids that increased competition among bacteria. Organic acid and polyphenols significantly modified rumen bacterial populations during high-grain feeding in dairy heifers. In particular the polyphenol treatment increased the richness and diversity of rumen microbiota, which are usually high in conditions of physiological rumen pH and rumen function.

  18. Simultaneous Amplicon Sequencing to Explore Co-Occurrence Patterns of Bacterial, Archaeal and Eukaryotic Microorganisms in Rumen Microbial Communities

    PubMed Central

    Kittelmann, Sandra; Seedorf, Henning; Walters, William A.; Clemente, Jose C.; Knight, Rob; Gordon, Jeffrey I.; Janssen, Peter H.

    2013-01-01

    Ruminants rely on a complex rumen microbial community to convert dietary plant material to energy-yielding products. Here we developed a method to simultaneously analyze the community's bacterial and archaeal 16S rRNA genes, ciliate 18S rRNA genes and anaerobic fungal internal transcribed spacer 1 genes using 12 DNA samples derived from 11 different rumen samples from three host species (Ovis aries, Bos taurus, Cervus elephas) and multiplex 454 Titanium pyrosequencing. We show that the mixing ratio of the group-specific DNA templates before emulsion PCR is crucial to compensate for differences in amplicon length. This method, in contrast to using a non-specific universal primer pair, avoids sequencing non-targeted DNA, such as plant- or endophyte-derived rRNA genes, and allows increased or decreased levels of community structure resolution for each microbial group as needed. Communities analyzed with different primers always grouped by sample origin rather than by the primers used. However, primer choice had a greater impact on apparent archaeal community structure than on bacterial community structure, and biases for certain methanogen groups were detected. Co-occurrence analysis of microbial taxa from all three domains of life suggested strong within- and between-domain correlations between different groups of microorganisms within the rumen. The approach used to simultaneously characterize bacterial, archaeal and eukaryotic components of a microbiota should be applicable to other communities occupying diverse habitats. PMID:23408926

  19. The Role of Ciliate Protozoa in the Rumen

    PubMed Central

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

    2015-01-01

    First described in 1843, Rumen protozoa with their striking appearance were assumed to be important for the welfare of their host. However, despite contributing up to 50% of the bio-mass in the rumen, the role of protozoa in rumen microbial ecosystem remains unclear. Phylogenetic analysis of 18S rDNA libraries generated from the rumen of cattle, sheep, and goats has revealed an unexpected diversity of ciliated protozoa although variation in gene copy number between species makes it difficult to obtain absolute quantification. Despite repeated attempts it has proven impossible to maintain rumen protozoa in axenic culture. Thus it has been difficult to establish conclusively a role of ciliate protozoa in rumen fiber degradation. The development of techniques to clone and express ciliate genes in λ phage, together with bioinformatic indices to confirm the ciliate origin of the genes has allowed the isolation and characterization of fibrolytic genes from rumen protozoa. Elimination of the ciliate protozoa increases microbial protein supply by up to 30% and reduces methane production by up to 11%. Our recent findings suggest that holotrich protozoa play a disproportionate role in supporting methanogenesis whilst the small Entodinium are responsible for much of the bacterial protein turnover. As yet no method to control protozoa in the rumen that is safe and practically applicable has been developed, however a range of plant extract capable of controlling if not completely eliminating rumen protozoa have been described. PMID:26635774

  20. The Role of Ciliate Protozoa in the Rumen.

    PubMed

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

    2015-01-01

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

  1. Rumen microbial diversity in Svalbard reindeer, with particular emphasis on methanogenic archaea.

    PubMed

    Sundset, Monica A; Edwards, Joan E; Cheng, Yan Fen; Senosiain, Roberto S; Fraile, Maria N; Northwood, Korinne S; Praesteng, Kirsti E; Glad, Trine; Mathiesen, Svein D; Wright, André-Denis G

    2009-12-01

    Ruminal methanogens, bacteria and ciliate protozoa of Svalbard reindeer grazing natural pastures in October (late fall) and April (late winter) were investigated using molecular-based approaches. The appetite of the Svalbard reindeer peaks in August (summer) and is at its lowest in March (winter). Microbial numbers, quantified by real-time PCR, did not change significantly between October and April, when food intakes are at similar levels, although the numbers of methanogens tended to be higher in October (P=0.074), and ciliate numbers tended to be higher in April (P=0.055). Similarly, no change was detected in the bacterial and protozoal population composition by rRNA gene-based denaturing gradient gel electrophoresis analysis. Dominant methanogens were identified using a 16S rRNA gene library (97 clones) prepared from pooled PCR products from reindeer on October pasture (n=5). Eleven of the 22 distinct operational taxonomic units (OTUs) generated exhibited a high degree of sequence similarity to methanogens affiliated with Methanobacteriales (eight OTUs), Methanomicrobiales (one OTU) and Methanosarcinales (two OTUs). The remaining 11 OTUs (53% of the clones) were associated with a cluster of uncultivated ruminal archaea. This study has provided important insights into the rumen microbiome of a high-arctic herbivorous animal living under harsh nutritional conditions, and evidence suggesting that host type affects the population size of ruminal methanogens.

  2. Analysis of rumen microbial populations in lactating dairy cattle fed diets varying in carbohydrate profiles and Saccharomyces cerevisiae fermentation product.

    PubMed

    Mullins, C R; Mamedova, L K; Carpenter, A J; Ying, Y; Allen, M S; Yoon, I; Bradford, B J

    2013-09-01

    The rumen microbial ecosystem is a critical factor that links diets to bovine physiology and productivity; however, information about dietary effects on microbial populations has generally been limited to small numbers of samples and qualitative assessment. To assess whether consistent shifts in microbial populations occur in response to common dietary manipulations in dairy cattle, samples of rumen contents were collected from 2 studies for analysis by quantitative real-time PCR (qPCR). In one study, lactating Holstein cows (n=8) were fed diets in which a nonforage fiber source replaced an increasing proportion of forages and concentrates in a 4×4 Latin square design, and samples of ruminal digesta were collected at 9-h intervals over 3 d at the end of each period. In the second study, lactating Holstein cows (n=15) were fed diets with or without the inclusion of a Saccharomyces cerevisiae fermentation product (SCFP) in a crossover design. In this study, rumen liquid and solid samples were collected during total rumen evacuations before and after feeding in a 42-h period. In total, 146 samples of ruminal digesta were used for microbial DNA isolation and analysis by qPCR. Validated primer sets were used to quantify total bacterial and anaerobic fungal populations as well as 12 well-studied bacterial taxa. The relative abundance of the target populations was similar to those previously reported. No significant treatment effects were observed for any target population. A significant interaction of treatment and dry matter intake was observed, however, for the abundance of Eubacterium ruminantium. Increasing dry matter intake was associated with a quadratic decrease in E. ruminantium populations in control animals but with a quadratic increase in E.ruminantium populations in cows fed SCFP. Analysis of sample time effects revealed that Fibrobacter succinogenes and fungal populations were more abundant postfeeding, whereas Ruminococcus albus tended to be more abundant

  3. Effect of monensin withdrawal on rumen fermentation, methanogenesis and microbial populations in cattle.

    PubMed

    Abrar, Arfan; Tsukahara, Takamitsu; Kondo, Makoto; Ban-Tokuda, Tomomi; Chao, Wang; Matsui, Hiroki

    2015-09-01

    This study was designed to obtain information on the residual influence of dietary monensin on ruminant fermentation, methanogenesis and bacterial population. Three ruminally cannulated crossbreed heifers (14 months old, 363 ± 11 kg) were fed Italian ryegrass straw and concentrate supplemented with monensin for 21 days before sampling. Rumen fluid samples were collected for analysis of short chain fatty acid (SCFA) profiles, monensin concentration, methanogens and rumen bacterial density. Post-feeding rumen fluid was also collected to determine in vitro gas production. Monensin was eliminated from the rumen fluid within 3 days. The composition of SCFA varied after elimination of monensin, while total production of SCFA was 1.78 times higher than on the first day. Methane production increased 7 days after monensin administration ceased, whereas hydrogen production decreased. The methanogens and rumen bacterial copy numbers were unaffected by the withdrawal of monensin. © 2015 Japanese Society of Animal Science.

  4. The degradation of different protein supplements in the rumen of steers and the effects of these supplements on carbohydrate digestion.

    PubMed

    McAllan, A B; Cockburn, J E; Williams, A P; Smith, R H

    1988-11-01

    1. Four steers with simple rumen and abomasal cannulas were given diets consisting of ground and pelleted alkali-treated straw, rolled barley and tapioca supplemented with urea (diet U) or containing single-cell protein (diet SCP), maize-gluten meal (diet MGM) or rapeseed meal (diet RSM) in place of some of the tapioca. The isoenergetic diets were given in a 4 x 4 Latin square design in eight feeds/d at 3-h intervals and provided sufficient metabolizable energy to support a growth rate of approximately 0.5 kg/d. Chromic oxide and polyethylene glycol were given as markers and appropriate samples taken from the rumen and abomasum. Flows (g/d) at the abomasum of organic matter and nitrogenous and carbohydrate constituents were calculated. 2. Rumen ammonia levels were similar with all three protein supplements at about 9 mmol/l, which was significantly lower (P less than 0.05) than that in animals on diet U (16 mmol/l). Rumen liquid outflow rates (/h) were 0.099, 0.139, 0.125 and 0.160 for diets U, SCP, MGM and RSM respectively; the difference between diet U and diet RSM was significant (P less than 0.05). Corresponding values for Cr2O3 outflow rates were 0.027, 0.032, 0.027 and 0.030/h respectively, which did not differ significantly from each other. 3. RNA, 35S and diaminopimelic acid (DAP) were used as microbial markers. Efficiencies of microbial-N (MN) synthesis, expressed as g MN/kg apparently digestible organic matter, truly digestible organic matter or carbohydrate fermented, were generally not significantly affected by the diet and averaged 29, 22 and 29 respectively based on mean RNA and 35S markers. Corresponding values derived from DAP of 22, 16 and 21 g MN/kg respectively were all significantly (P less than 0.001) lower. Using 35S as microbial marker, MN flows at the abomasum as a proportion of non-ammonia-nitrogen flow were 0.78, 0.64, 0.51 and 0.78 for diets U, SCP, MGM and RSM respectively. Derived true rumen degradability values (g/g intake) of the

  5. The Contribution of Mathematical Modeling to Understanding Dynamic Aspects of Rumen Metabolism

    PubMed Central

    Bannink, André; van Lingen, Henk J.; Ellis, Jennifer L.; France, James; Dijkstra, Jan

    2016-01-01

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

  6. Molecular comparative assessment of the microbial ecosystem in rumen and faeces of goats fed alfalfa hay alone or combined with oats.

    PubMed

    Mohammadzadeh, Hamid; Yáñez-Ruiz, David R; Martínez-Fernandez, Gonzalo; Abecia, Leticia

    2014-10-01

    The objective of this work was to compare the biomass and community structure of bacteria, protozoa and archaea communities in samples of rumen and faeces of goats and to what extent the diet (alfalfa hay with or without supplemented oats) offered to them exert an influence. Four cannulated adult goats fistulated in the rumen were used in a cross over design experiment in two experimental periods of 26 days, consisting in 14 days of adaptation, 7 days of sampling rumen contents and 5 days of digestibility measurement. Bacterial, protozoa and archaeal biomass and the communities' structure was assessed by real time PCR (qPCR) and denaturing gradient gel electrophoresis (DGGE), respectively. The numbers of archaea and bacteria in both rumen and faeces were higher and lower, respectively, in animals fed AH diet (P < 0.005). Contrary, protozoal numbers were not affected by the diet but were lower (P < 0.001) in faeces than in rumen. The analysis of the community structure revealed a consistently different population in structure in rumen and faeces for the three studied microbial groups and that supplementing alfalfa hay with oats led to a decrease in the similarity between sites in the rumen and faeces: similarity indexes for bacteria (57 and 27%), archaea (26 and 9%) and protozoa (62 and 22%) in animals fed AH and AHO diets, respectively.

  7. Effect of DNA Extraction Methods on the Apparent Structure of Yak Rumen Microbial Communities as Revealed by 16S rDNA Sequencing.

    PubMed

    Chen, Ya-Bing; Lan, Dao-Liang; Tang, Cheng; Yang, Xiao-Nong; Li, Jian

    2015-01-01

    To more efficiently identify the microbial community of the yak rumen, the standardization of DNA extraction is key to ensure fidelity while studying environmental microbial communities. In this study, we systematically compared the efficiency of several extraction methods based on DNA yield, purity, and 16S rDNA sequencing to determine the optimal DNA extraction methods whose DNA products reflect complete bacterial communities. The results indicate that method 6 (hexadecyltrimethylammomium bromide-lysozyme-physical lysis by bead beating) is recommended for the DNA isolation of the rumen microbial community due to its high yield, operational taxonomic unit, bacterial diversity, and excellent cell-breaking capability. The results also indicate that the bead-beating step is necessary to effectively break down the cell walls of all of the microbes, especially Gram-positive bacteria. Another aim of this study was to preliminarily analyze the bacterial community via 16S rDNA sequencing. The microbial community spanned approximately 21 phyla, 35 classes, 75 families, and 112 genera. A comparative analysis showed some variations in the microbial community between yaks and cattle that may be attributed to diet and environmental differences. Interestingly, numerous uncultured or unclassified bacteria were found in yak rumen, suggesting that further research is required to determine the specific functional and ecological roles of these bacteria in yak rumen. In summary, the investigation of the optimal DNA extraction methods and the preliminary evaluation of the bacterial community composition of yak rumen support further identification of the specificity of the rumen microbial community in yak and the discovery of distinct gene resources.

  8. Influence of Sire Breed on the Interplay among Rumen Microbial Populations Inhabiting the Rumen Liquid of the Progeny in Beef Cattle

    PubMed Central

    Hernandez-Sanabria, Emma; Goonewardene, Laksiri A.; Wang, Zhiquan; Zhou, Mi; Moore, Stephen S.; Guan, Le Luo

    2013-01-01

    This study aimed to evaluate whether the host genetic background impact the ruminal microbial communities of the progeny of sires from three different breeds under different diets. Eighty five bacterial and twenty eight methanogen phylotypes from 49 individuals of diverging sire breed (Angus, ANG; Charolais, CHA; and Hybrid, HYB), fed high energy density (HE) and low energy density (LE) diets were determined and correlated with breed, rumen fermentation and phenotypic variables, using multivariate statistical approaches. When bacterial phylotypes were compared between diets, ANG offspring showed the lowest number of diet-associated phylotypes, whereas CHA and HYB progenies had seventeen and twenty-three diet-associated phylotypes, respectively. For the methanogen phylotypes, there were no sire breed-associated phylotypes; however, seven phylotypes were significantly different among breeds on either diet (P<0.05). Sire breed did not influence the metabolic variables measured when high energy diet was fed. A correlation matrix of all pairwise comparisons among frequencies of bacterial and methanogen phylotypes uncovered their relationships with sire breed. A cluster containing methanogen phylotypes M16 (Methanobrevibacter gottschalkii) and M20 (Methanobrevibacter smithii), and bacterial phylotype B62 (Robinsoniella sp.) in Angus offspring fed low energy diet reflected the metabolic interactions among microbial consortia. The clustering of the phylotype frequencies from the three breeds indicated that phylotypes detected in CHA and HYB progenies are more similar among them, compared to ANG animals. Our results revealed that the frequency of particular microbial phylotypes in the progeny of cattle may be influenced by the sire breed when different diets are fed and ultimately further impact host metabolic functions, such as feed efficiency. PMID:23520513

  9. Distribution and Genetic Diversity of Bacteriocin Gene Clusters in Rumen Microbial Genomes

    PubMed Central

    Azevedo, Analice C.; Bento, Cláudia B. P.; Ruiz, Jeronimo C.; Queiroz, Marisa V.

    2015-01-01

    Some species of ruminal bacteria are known to produce antimicrobial peptides, but the screening procedures have mostly been based on in vitro assays using standardized methods. Recent sequencing efforts have made available the genome sequences of hundreds of ruminal microorganisms. In this work, we performed genome mining of the complete and partial genome sequences of 224 ruminal bacteria and 5 ruminal archaea to determine the distribution and diversity of bacteriocin gene clusters. A total of 46 bacteriocin gene clusters were identified in 33 strains of ruminal bacteria. Twenty gene clusters were related to lanthipeptide biosynthesis, while 11 gene clusters were associated with sactipeptide production, 7 gene clusters were associated with class II bacteriocin production, and 8 gene clusters were associated with class III bacteriocin production. The frequency of strains whose genomes encode putative antimicrobial peptide precursors was 14.4%. Clusters related to the production of sactipeptides were identified for the first time among ruminal bacteria. BLAST analysis indicated that the majority of the gene clusters (88%) encoding putative lanthipeptides contained all the essential genes required for lanthipeptide biosynthesis. Most strains of Streptococcus (66.6%) harbored complete lanthipeptide gene clusters, in addition to an open reading frame encoding a putative class II bacteriocin. Albusin B-like proteins were found in 100% of the Ruminococcus albus strains screened in this study. The in silico analysis provided evidence of novel biosynthetic gene clusters in bacterial species not previously related to bacteriocin production, suggesting that the rumen microbiota represents an underexplored source of antimicrobial peptides. PMID:26253660

  10. Thermodynamic Driving Force of Hydrogen on Rumen Microbial Metabolism: A Theoretical Investigation.

    PubMed

    van Lingen, Henk J; Plugge, Caroline M; Fadel, James G; Kebreab, Ermias; Bannink, André; Dijkstra, Jan

    2016-01-01

    Hydrogen is a key product of rumen fermentation and has been suggested to thermodynamically control the production of the various volatile fatty acids (VFA). Previous studies, however, have not accounted for the fact that only thermodynamic near-equilibrium conditions control the magnitude of reaction rate. Furthermore, the role of NAD, which is affected by hydrogen partial pressure (PH2), has often not been considered. The aim of this study was to quantify the control of PH2 on reaction rates of specific fermentation pathways, methanogenesis and NADH oxidation in rumen microbes. The control of PH2 was quantified using the thermodynamic potential factor (FT), which is a dimensionless factor that corrects a predicted kinetic reaction rate for the thermodynamic control exerted. Unity FT was calculated for all glucose fermentation pathways considered, indicating no inhibition of PH2 on the production of a specific type of VFA (e.g., acetate, propionate and butyrate) in the rumen. For NADH oxidation without ferredoxin oxidation, increasing PH2 within the rumen physiological range decreased FT from unity to zero for different NAD+ to NADH ratios and pH of 6.2 and 7.0, which indicates thermodynamic control of PH2. For NADH oxidation with ferredoxin oxidation, increasing PH2 within the rumen physiological range decreased FT from unity at pH of 7.0 only. For the acetate to propionate conversion, FT increased from 0.65 to unity with increasing PH2, which indicates thermodynamic control. For propionate to acetate and butyrate to acetate conversions, FT decreased to zero below the rumen range of PH2, indicating full thermodynamic suppression. For methanogenesis by archaea without cytochromes, FT differed from unity only below the rumen range of PH2, indicating no thermodynamic control. This theoretical investigation shows that thermodynamic control of PH2 on individual VFA produced and associated yield of hydrogen and methane cannot be explained without considering NADH

  11. Thermodynamic Driving Force of Hydrogen on Rumen Microbial Metabolism: A Theoretical Investigation

    PubMed Central

    van Lingen, Henk J.; Plugge, Caroline M.; Fadel, James G.; Kebreab, Ermias; Bannink, André; Dijkstra, Jan

    2016-01-01

    Hydrogen is a key product of rumen fermentation and has been suggested to thermodynamically control the production of the various volatile fatty acids (VFA). Previous studies, however, have not accounted for the fact that only thermodynamic near-equilibrium conditions control the magnitude of reaction rate. Furthermore, the role of NAD, which is affected by hydrogen partial pressure (PH2), has often not been considered. The aim of this study was to quantify the control of PH2 on reaction rates of specific fermentation pathways, methanogenesis and NADH oxidation in rumen microbes. The control of PH2 was quantified using the thermodynamic potential factor (FT), which is a dimensionless factor that corrects a predicted kinetic reaction rate for the thermodynamic control exerted. Unity FT was calculated for all glucose fermentation pathways considered, indicating no inhibition of PH2 on the production of a specific type of VFA (e.g., acetate, propionate and butyrate) in the rumen. For NADH oxidation without ferredoxin oxidation, increasing PH2 within the rumen physiological range decreased FT from unity to zero for different NAD+ to NADH ratios and pH of 6.2 and 7.0, which indicates thermodynamic control of PH2. For NADH oxidation with ferredoxin oxidation, increasing PH2 within the rumen physiological range decreased FT from unity at pH of 7.0 only. For the acetate to propionate conversion, FT increased from 0.65 to unity with increasing PH2, which indicates thermodynamic control. For propionate to acetate and butyrate to acetate conversions, FT decreased to zero below the rumen range of PH2, indicating full thermodynamic suppression. For methanogenesis by archaea without cytochromes, FT differed from unity only below the rumen range of PH2, indicating no thermodynamic control. This theoretical investigation shows that thermodynamic control of PH2 on individual VFA produced and associated yield of hydrogen and methane cannot be explained without considering NADH

  12. Changes in Rumen Microbial Community Composition during Adaption to an In Vitro System and the Impact of Different Forages

    PubMed Central

    Lengowski, Melanie B.; Zuber, Karin H. R.; Witzig, Maren; Möhring, Jens; Boguhn, Jeannette; Rodehutscord, Markus

    2016-01-01

    This study examined ruminal microbial community composition alterations during initial adaption to and following incubation in a rumen simulation system (Rusitec) using grass or corn silage as substrates. Samples were collected from fermenter liquids at 0, 2, 4, 12, 24, and 48 h and from feed residues at 0, 24, and 48 h after initiation of incubation (period 1) and on day 13 (period 2). Microbial DNA was extracted and real-time qPCR was used to quantify differences in the abundance of protozoa, methanogens, total bacteria, Fibrobacter succinogenes, Ruminococcus albus, Ruminobacter amylophilus, Prevotella bryantii, Selenomonas ruminantium, and Clostridium aminophilum. We found that forage source and sampling time significantly influenced the ruminal microbial community. The gene copy numbers of most microbial species (except C. aminophilum) decreased in period 1; however, adaption continued through period 2 for several species. The addition of fresh substrate in period 2 led to increasing copy numbers of all microbial species during the first 2–4 h in the fermenter liquid except protozoa, which showed a postprandial decrease. Corn silage enhanced the growth of R. amylophilus and F. succinogenes, and grass silage enhanced R. albus, P. bryantii, and C. aminophilum. No effect of forage source was detected on total bacteria, protozoa, S. ruminantium, or methanogens or on total gas production, although grass silage enhanced methane production. This study showed that the Rusitec provides a stable system after an adaption phase that should last longer than 48 h, and that the forage source influenced several microbial species. PMID:26928330

  13. Effects of supplementing an active dry yeast product on rumen microbial community composition and on subsequent rumen fermentation of lactating cows in the mid-to-late lactation period.

    PubMed

    Uyeno, Yutaka; Akiyama, Kiyoshi; Hasunuma, Toshiya; Yamamoto, Hiroshi; Yokokawa, Hiroaki; Yamaguchi, Tsuneko; Kawashima, Kenji; Itoh, Minoru; Kushibiki, Shiro; Hirako, Makoto

    2017-01-01

    The effects of supplementing feed of cows in mid-to-late lactation with an active yeast product (Actisaf Sc 47) were evaluated using 15 Holstein cows in a replicated 3 × 3 Latin square design. The animals were fed a mixed ration with 33% neutral detergent fiber, consisting of timothy hay (29.8%), a commercial concentrate (70.0%) and commercial calcium triphosphate (0.2%), twice daily to meet 105% of their energy requirement. Yeast supplement was set at 0, 5 and 10 g per day over 21-day periods, each of which consisted of 14 days for adaptation followed by 7 days of data collection. Milking performance, plasma metabolite parameters, rumen volatile fatty acids, lipopolysaccharide and microbial properties were measured. Although there were no significant differences in feeding and milking performance or blood parameters associated with supplementation, the acetate to propionate ratio in the rumen fluid tended to decrease (P = 0.08). The population of Bacteroidetes tended to be less prominent (P = 0.07) and the fibrolytic bacterium Fibrobacter significantly increased (P < 0.05) in the rumen fluid of the yeast 10 g group compared with that of the control. These data suggest that effects of supplementing live yeast to cows in mid-to-late lactation may be limited to microbial composition and fermentation characteristics in the rumen. © 2016 Japanese Society of Animal Science.

  14. Effects of Phytoecdysteroids (PEDS) Extracted from Cyanotis arachnoidea on Rumen Fermentation, Enzyme Activity and Microbial Efficiency in a Continuous-Culture System

    PubMed Central

    Li, Deyong; Zhang, Yawei; Cui, Zhenliang; He, Liwen; Chen, Wanbao; Meng, Qingxiang; Ren, Liping

    2016-01-01

    The objective of this study was to evaluate the effects of supplementation of phytoecdysteroids (PEDS) extracted from Cyanotis arachnoidea on rumen fermentation, enzymes activity and microbial efficiency in a dual flow continuous-culture system. A single-factor experimental design was used with twelve fermenters in 4 groups with 3 replicates each. Fermenters were incubated for a total of 7 days that included first 4 days for adaptation and last 3 days for sampling. PEDS was added at levels of zero (as control), 5, 10, and 15 mg/g of the substrate (DM). The results showed that increasing supplementation levels of PEDS resulted in incremental digestibility of dry matter (DMD) (quadratic, P = 0.001) and organic matter (OMD) (quadratic, P = 0.031), but unchanged digestibility of neutral detergent fiber (NDFD), crude protein (CPD) and acid detergent acid (ADFD). As supplementation levels of PEDS increased, there were decreased response in the concentration of ammonia nitrogen (NH3-N) (linear, P = 0.015) and increased response in molar proportions of butyrate (linear, P = 0.004), but unchanged response in total volatile fatty acid (TVFA) and the molar proportion of acetate and propionate, respectively. Increasing PEDS supplementation levels decreased the ratio of acetate to propionate (linear, P = 0.038), suggesting an alteration of rumen fermentation pattern occurring due to PEDS supplementation in the diet. Supplementation of PEDS significantly increased activities of glutamate dehydrogenase (quadratic, P = 0.001), alanine dehydrogenase (quadratic, P = 0.004), glutamate synthase (linear, P = 0.038), glutamine synthetase (quadratic, P = 0.011), respectively. There were no discernible differences in the activity of carboxymethyl cellulose (CMCase), xylanase and protease regardless of the treatments. The daily production of microbial nitrogen (linear, P = 0.002) and microbial efficiency (MOEEF) (linear, P = 0.001) increased linearly as supplementation levels of PEDS

  15. A novel feruloyl esterase from rumen microbial metagenome: Gene cloning and enzyme characterization in the release of mono- and diferulic acids

    USDA-ARS?s Scientific Manuscript database

    A feruloyl esterase (FAE) gene was isolated from a rumen microbial metagenome, cloned into E. coli, and expressed in active form. The enzyme (RuFae4) was classified as a Type D feruloyl esterase based on its action on synthetic substrates and ability to release diferulates. The RuFae4 alone releas...

  16. Effects of rumen fluid collection site on microbial population structure during in vitro fermentation of the different substrates quantified by 16S rRNA hybridisation.

    PubMed

    Muetzel, S; Krishnamoorthy, U; Becker, K

    2001-01-01

    Rumen fluid samples from a cow were withdrawn manually from the feed mat (solid phase) or the liquid phase below this mat and incubated in vitro with wheat straw, sorghum hay and a concentrate mixture. From the inoculum and several samples collected during in vitro incubation RNA was extracted to assess microbial population size and structure. RNA content recovered from the solid phase rumen fluid was significantly higher than from the liquid phase. The composition of the microbial population in the solid phase material was characterised by a high proportion of Ruminococci. Neither the proportion of other cell wall degrading organisms (Fibrobacter and Chytridiomycetes) nor the Eukarya and Archaea populations differed between the two sampling sites. Gas production was higher when substrates were incubated with solid phase than with liquid phase rumen fluid regardless of sampling time. However, the higher level of gas production was not accompanied by a corresponding increase in true digestibility. The RNA probes showed that during in vitro incubation with liquid phase rumen fluid, the eukaryotic population was inactive no matter which substrate was used and the activity of methanogens (Archaea) was lower than with solid phase rumen fluid. The population pattern of the cell wall degrading organisms was influenced mainly by the substrate fermented, and to a smaller extent by the inoculum used for in vitro fermentation.

  17. Rumen fermentation and microbial population in lactating dairy cows receiving diets containing oilseeds rich in C-18 fatty acids.

    PubMed

    Ivan, M; Petit, H V; Chiquette, J; Wright, A-D G

    2013-04-14

    Sixteen Holstein rumen-cannulated primiparous milking dairy cows were fed a control diet (CN) based on maize silage and soyabean meal during a 4-week period before the start of a 21-d experiment with oilseeds containing high concentration of linoleic acid (Linola™) or linolenic acid (NuLin™). Thereafter, four cows received ad libitum one of each of four dietary treatments comprising of CN, Linola (LN), NuLin (NL) and LN/NL (50/50 % combination). Each LN, NL and LN/NL treatment contained 6 % oil of DM. Rumen digesta samples were collected on days 6, 11, 16 and 21 and milk samples on days 13, 15 and 17. There were no effects (P>0.05) of the oilseeds on pH and concentrations of NH3-N and total volatile fatty acids, while the acetate:propionate ratio was decreased (P< 0.05). The oilseeds also decreased (P< 0.05) protozoa and increased (P< 0.1) total cellulolytic bacteria in rumen fluid, especially when containing high dietary linoleic acid (P< 0.05). The milk protein concentration was increased (P< 0.1) by the dietary linoleic acid, which produced most beneficial results. It was concluded that supplements of linoleic acid in diets of ruminants might contribute to better digestion of dietary fibre and increased quality of milk.

  18. Impact of dietary rumen undegradable protein and rumen-protected choline on intake, peripartum liver triacylglyceride, plasma metabolites and milk production in transition dairy cows.

    PubMed

    Hartwell, J R; Cecava, M J; Donkin, S S

    2000-12-01

    The objectives of the present study were to determine the effects of rumen undegradable protein (RUP) level of prepartum diets, the supplementation of a rumen-protected choline product, and their interactions on milk production, feed intake, body weight and condition, blood metabolites, and liver triacylglycerides in dairy cows. Rumen-protected choline (RPC) was fed with two levels of RUP to 48 multiparous Holstein cows in a 3 x 2 factorial arrangement of treatments. Beginning 28 d before expected calving, cows were fed 10% rumen degradable protein, either 0, 6, or 12 g/d of RPC as CapShure (Balchem Corp., Slate Hill, NY) and either 4.0 or 6.2% RUP. After calving and through 120 d of lactation, cows received a common diet and continued RPC as per their prepartum assignment. Prepartum dry matter intake (kg/d) was not affected by RPC or RUP. Postpartum intake decreased when 6.2% RUP was fed prepartum. Milk production to 56 d in milk was decreased when cows were fed 6.2% RUP prepartum. Milk protein (kg/d) decreased when additional RUP was fed prepartum. Cows fed RPC lost more weight during the study period and tended to lose more body condition. Plasma urea nitrogen levels in the prepartum period were reduced for cows fed 4.0% RUP prepartum. Mean liver triacylglyceride determined from samples obtained at -28, -14, +1, +28, and +56 d in milk was not affected by RPC, prepartum RUP, or their combinations. Feeding 12 g of RPC/d in conjunction with 4.0% RUP increased milk production, but feeding RPC with 6.2% RUP prepartum and through 56 d in milk decreased production. The data indicate that 6.2% RUP does not benefit close-up dry cows, and the response to RPC depends the RUP content of the prepartum diet.

  19. Effects of protein sources on concentrations of hydrogen sulphide in the rumen headspace gas of dairy cows.

    PubMed

    Fonseca, A J M; Cabrita, A R J; Pinho, L A O; Kim, E J; Dewhurst, R J

    2013-01-01

    Two Latin square design experiments investigated the relationship between hydrogen sulphide concentration in the rumen headspace gas of dairy cows and the early stages of protein degradation in the rumen. In Expt 1, three protein sources differing in rumen N (nitrogen) degradability (maize gluten feed (MGF); sunflower meal (SFM); and soyabean meal (SBM)) were used, whereas in Expt 2 four different batches of the same feed (MGF) differing in colour (CIE L*, a*, b* (CIELAB) scale) were used. After allowing the concentration of hydrogen sulphide in rumen gas to decline close to zero, a fixed amount of protein sources was offered to cows and the concentrations of hydrogen sulphide were recorded in rumen headspace gas at 30-min intervals. In Expt 1, the concentration of hydrogen sulphide showed considerable variation between protein sources, with MGF having the highest concentration followed by SFM and SBM resulting in very low concentrations. The N wash losses (zero time measurements with nylon bags) ranked the feeds in the same way, from MGF (highest; 61%) to SBM (lowest; 26%). There were marked differences in the degradation of cystine and methionine between protein sources, although the degradation of cystine was always higher than for methionine. MGF (Expt 2) led to increased concentrations of hydrogen sulphide, with peak concentrations achieved between 1 and 2 h after feeding. The concentrations of hydrogen sulphide were higher for MGF1, intermediate for MGF2 and lower for MGF3 and MGF4, agreeing with colour scale. Differences in the early stages of dietary sulphur degradation corresponded with differences in hydrogen sulphide concentrations in rumen gas. The results suggest that hydrogen sulphide concentrations in the rumen headspace gas could be useful to evaluate nutritional parameters not measured by the in sacco technique, contributing to a better understanding of the response of dairy cows to different protein supplements.

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

    PubMed

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

    2013-05-01

    The methodological procedures for studying the fibrolytic activity of rumen bacteria are not clearly established. In this study the efficiency of sonication treatment and buffer composition (i.e. buffer varying in tonicity or pH) on the level of protein extraction from the residue of forage samples incubated in the rumen of a grazing steer and the effect of buffer composition or CaCl₂ concentration on the carboxymethylcellulase (CMCase) activity of the released protein were evaluated. The amount of protein released from the residue of incubation was higher (P < 0.05) for the sonicated material and increased linearly with increasing buffer pH (P < 0.05). The CMCase activity of the released protein was not improved by sonication treatment, whereas it was higher (P < 0.05) for hypotonic than for hypertonic buffer. Both linear and quadratic effects (P < 0.05) of buffer pH on CMCase activity were significant, with CMCase activity being maximal at pH 5.4-6.1. CMCase activity was higher (P < 0.05) at a CaCl₂ concentration of 1 mmol L(-1) compared with lower values. Although sonication treatment increases the amount of protein extracted from rumen bacteria adhered to the residue of incubation, the CMCase activity of the released protein might be measured without sonication treatment and should be carried out with a hypotonic buffer solution that includes a calcium source. When pH is not a treatment factor, the buffer pH should be between 5.5 and 6. © 2012 Society of Chemical Industry.

  1. Effect of dietary level of rumen-degraded protein on production and nitrogen metabolism in lactating dairy cows.

    PubMed

    Reynal, S M; Broderick, G A

    2005-11-01

    Twenty-eight (8 with ruminal cannulas) lactating Holstein cows were assigned to 4 x 4 Latin squares and fed diets with different levels of rumen-degraded protein (RDP) to study the effect of RDP on production and N metabolism. Diets contained [dry matter (DM) basis] 37% corn silage, 13% alfalfa silage, and 50% concentrate. The concentrate contained solvent and lignosulfonate-treated soybean meal and urea, and was adjusted to provide RDP at: 13.2, 12.3, 11.7, and 10.6% of DM in diets A to D, respectively. Intake of DM and yield of milk, fat-corrected milk, and fat were not affected by treatments. Dietary RDP had positive linear effects on milk true protein content and microbial non-ammonia N (NAN) flow at the omasal canal, and a quadratic effect on true protein yield, with maximal protein production at 12.3% RDP. However, dietary RDP had a positive linear effect on total N excretion, with urinary N accounting for most of the increase, and a negative linear effect on environmental N efficiency (kg of milk produced per kg of N excreted). Therefore, a compromise between profitability and environmental quality was achieved at a dietary RDP level of 11.7% of DM. Observed microbial NAN flow and RDP supply were higher and RUP flow was lower than those predicted by the NRC (2001) model. The NRC (2001) model overpredicted production responses to RUP compared with the results in this study. Replacing default NRC degradation rates for protein supplements with rates measured in vivo resulted in similar observed and predicted values, suggesting that in situ degradation rates used by the NRC are slower than apparent rates in this study.

  2. Effects of rare earth element lanthanum on rumen methane and volatile fatty acid production and microbial flora in vitro.

    PubMed

    Zhang, T T; Zhao, G Y; Zheng, W S; Niu, W J; Wei, C; Lin, S X

    2015-06-01

    The objectives of the trial were to study the effects of rare earth element (REE) lanthanum (La) on the in vitro rumen methane (CH4 ) and volatile fatty acid (VFA) production and the microbial flora of feeds. Four feed mixtures with different levels of neutral detergent fibre (NDF), that is 20.0% (I), 31.0% (II), 41.9% (III) and 52.7% (IV), were formulated as substrates. Five levels of LaCl3 , that is 0, 0.4, 0.6, 0.8 and 1.0 mmol/kg dry matter (DM), were added to the feed mixtures, respectively, as experimental treatments in a two-factor 5 × 4 randomized design. The in vitro incubation lasted for 24 h. The results showed that supplementing LaCl3 increased the total gas (p < 0.001) production and tended to increase the total VFA production (p = 0.072) and decreased the CH4 production (p = 0.001) and the ratios of acetate/propionate (p = 0.019) and CH4 /total VFA (p < 0.001). Interactions between LaCl3 and NDF were significant in total gas production (p = 0.030) and tended to be significant in CH4 production (p = 0.071). Supplementing LaCl3 at the level of 0.8 mmol/g DM decreased the relative abundance of methanogens and protozoa in the total bacterial 16S rDNA analysed using the real-time PCR (p < 0.0001), increased F. succinogenes (p = 0.0003) and decreased R. flavefaciens (p < 0.0001) whereas did not affect R. albus and anaerobic fungi (p > 0.05). It was concluded that LaCl3 decreased the CH4 production without negatively affecting feed digestion through manipulating rumen microbial flora when feed mixtures with different levels of NDF were used as substrates.

  3. In vitro-in vivo study on the effects of plant compounds on rumen fermentation, microbial abundances and methane emissions in goats.

    PubMed

    Martínez-Fernández, G; Abecia, L; Martín-García, A I; Ramos-Morales, E; Hervás, G; Molina-Alcaide, E; Yáñez-Ruiz, D R

    2013-12-01

    Two in vitro and one in vivo experiments were conducted to investigate the effects of a selection of plant compounds on rumen fermentation, microbial concentration and methane emissions in goats. Treatments were: control (no additive), carvacrol (CAR), cinnamaldehyde (CIN), eugenol (EUG), propyl propane thiosulfinate (PTS), propyl propane thiosulfonate (PTSO), diallyl disulfide (DDS), a mixture (40 : 60) of PTS and PTSO (PTS+PTSO), and bromochloromethane (BCM) as positive control with proven antimethanogenic effectiveness. Four doses (40, 80, 160 and 320 µl/l) of the different compounds were incubated in vitro for 24 h in diluted rumen fluid from goats using two diets differing in starch and protein source within the concentrate (Experiment 1).The total gas production was linearly decreased (P<0.012) by all compounds, with the exception of EUG and PTS+PTSO (P≥ 0.366). Total volatile fatty-acid (VFA) concentration decreased (P≤ 0.018) only with PTS, PTSO and CAR, whereas the acetate:propionate ratio decreased (P≤ 0.002) with PTS, PTSO and BCM, and a tendency (P=0.064) was observed for DDS. On the basis of results from Experiment 1, two doses of PTS, CAR, CIN, BCM (160 and 320 µl/l), PTSO (40 and 160 µl/l) and DDS (80 and 320 µl/l) were further tested in vitro for 72 h (Experiment 2). The gas production kinetics were affected (P≤ 0.045) by all compounds, and digested NDF (DNDF) after 72 h of incubation was only linearly decreased (P≤ 0.004) by CAR and PTS. The addition of all compounds linearly decreased (P≤ 0.009) methane production, although the greatest reductions were observed for PTS (up to 96%), DDS (62%) and BCM (95%). No diet-dose interaction was observed. To further test the results obtained in vitro, two groups of 16 adult non-pregnant goats were used to study in vivo the effect of adding PTS (50, 100 and 200 mg/l rumen content per day) and BCM (50, 100 and 160 mg/l rumen content per day) during the 9 days on methane emissions (Experiment 3

  4. Bovine Host Genetic Variation Influences Rumen Microbial Methane Production with Best Selection Criterion for Low Methane Emitting and Efficiently Feed Converting Hosts Based on Metagenomic Gene Abundance.

    PubMed

    Roehe, Rainer; Dewhurst, Richard J; Duthie, Carol-Anne; Rooke, John A; McKain, Nest; Ross, Dave W; Hyslop, Jimmy J; Waterhouse, Anthony; Freeman, Tom C; Watson, Mick; Wallace, R John

    2016-02-01

    Methane produced by methanogenic archaea in ruminants contributes significantly to anthropogenic greenhouse gas emissions. The host genetic link controlling microbial methane production is unknown and appropriate genetic selection strategies are not developed. We used sire progeny group differences to estimate the host genetic influence on rumen microbial methane production in a factorial experiment consisting of crossbred breed types and diets. Rumen metagenomic profiling was undertaken to investigate links between microbial genes and methane emissions or feed conversion efficiency. Sire progeny groups differed significantly in their methane emissions measured in respiration chambers. Ranking of the sire progeny groups based on methane emissions or relative archaeal abundance was consistent overall and within diet, suggesting that archaeal abundance in ruminal digesta is under host genetic control and can be used to genetically select animals without measuring methane directly. In the metagenomic analysis of rumen contents, we identified 3970 microbial genes of which 20 and 49 genes were significantly associated with methane emissions and feed conversion efficiency respectively. These explained 81% and 86% of the respective variation and were clustered in distinct functional gene networks. Methanogenesis genes (e.g. mcrA and fmdB) were associated with methane emissions, whilst host-microbiome cross talk genes (e.g. TSTA3 and FucI) were associated with feed conversion efficiency. These results strengthen the idea that the host animal controls its own microbiota to a significant extent and open up the implementation of effective breeding strategies using rumen microbial gene abundance as a predictor for difficult-to-measure traits on a large number of hosts. Generally, the results provide a proof of principle to use the relative abundance of microbial genes in the gastrointestinal tract of different species to predict their influence on traits e.g. human metabolism

  5. Bovine Host Genetic Variation Influences Rumen Microbial Methane Production with Best Selection Criterion for Low Methane Emitting and Efficiently Feed Converting Hosts Based on Metagenomic Gene Abundance

    PubMed Central

    Roehe, Rainer; Dewhurst, Richard J.; Duthie, Carol-Anne; Rooke, John A.; McKain, Nest; Ross, Dave W.; Hyslop, Jimmy J.; Waterhouse, Anthony; Freeman, Tom C.

    2016-01-01

    Methane produced by methanogenic archaea in ruminants contributes significantly to anthropogenic greenhouse gas emissions. The host genetic link controlling microbial methane production is unknown and appropriate genetic selection strategies are not developed. We used sire progeny group differences to estimate the host genetic influence on rumen microbial methane production in a factorial experiment consisting of crossbred breed types and diets. Rumen metagenomic profiling was undertaken to investigate links between microbial genes and methane emissions or feed conversion efficiency. Sire progeny groups differed significantly in their methane emissions measured in respiration chambers. Ranking of the sire progeny groups based on methane emissions or relative archaeal abundance was consistent overall and within diet, suggesting that archaeal abundance in ruminal digesta is under host genetic control and can be used to genetically select animals without measuring methane directly. In the metagenomic analysis of rumen contents, we identified 3970 microbial genes of which 20 and 49 genes were significantly associated with methane emissions and feed conversion efficiency respectively. These explained 81% and 86% of the respective variation and were clustered in distinct functional gene networks. Methanogenesis genes (e.g. mcrA and fmdB) were associated with methane emissions, whilst host-microbiome cross talk genes (e.g. TSTA3 and FucI) were associated with feed conversion efficiency. These results strengthen the idea that the host animal controls its own microbiota to a significant extent and open up the implementation of effective breeding strategies using rumen microbial gene abundance as a predictor for difficult-to-measure traits on a large number of hosts. Generally, the results provide a proof of principle to use the relative abundance of microbial genes in the gastrointestinal tract of different species to predict their influence on traits e.g. human metabolism

  6. Strong associations among rumen endotoxin and acute phase proteins with plasma minerals in lactating cows fed graded amounts of concentrate.

    PubMed

    Zebeli, Q; Dunn, S M; Ametaj, B N

    2010-04-01

    The objective of this investigation was to determine associations among rumen endotoxin, plasma serum amyloid A (SAA), and C-reactive protein (CRP) with plasma Ca, Fe, Zn, and Cu in lactating cows challenged with graded amounts of rolled barley grain in the diet (i.e., 0, 15, 30, and 45% of DMI). Correlative relationships among variables were determined by linear and nonlinear regression procedures adjusted for the effects of day, animal, and experimental period. Increasing the amount of grain in the diet was successful in inducing an acute phase response, as assessed by augmentation of rumen endotoxin and plasma CRP and SAA (P < 0.01). The correlative analysis revealed inverse, nonlinear relationships of rumen endotoxin and plasma SAA with circulating Ca. Interestingly, plasma Ca reached the asymptotic plateau at 10.6 mg/dL. The increase in rumen endotoxin was associated with an abrupt decrease in plasma Fe (R(2) = 0.91; P < 0.001). A similar relationship, although at a reduced estimation accuracy (R(2) = 0.21; P < 0.01), was observed between rumen endotoxin and plasma Zn. Augmentation of rumen endotoxin and plasma CRP resulted in a positive, biphasic response of plasma Cu. In conclusion, the increase in rumen endotoxin in response to high-grain diets, and the resulting increases in plasma SAA and CRP, were strongly correlated with fluctuations of plasma minerals. Results suggest that new feeding strategies should be developed to curb the release of endotoxin in the rumen fluid to prevent perturbing minerals in the plasma.

  7. Effect of chestnut tannin on fermentation quality, proteolysis, and protein rumen degradability of alfalfa silage.

    PubMed

    Tabacco, E; Borreani, G; Crovetto, G M; Galassi, G; Colombo, D; Cavallarin, L

    2006-12-01

    Two experiments were conducted on alfalfa to investigate the effects of the addition of commercial chestnut hydrolyzable tannin at ensiling on 1) silage fermentation quality in lab-scale silos and protein degradation in the rumen, and 2) silage fermentation quality and proteolysis in bale silages. Wilted alfalfa was prepared with 4 tannin levels (0, 2, 4, and 6% on a dry matter (DM) basis; T0, T1, T2, T3, respectively) and ensiled in lab-scale silos. Silages (33% DM) were analyzed for fermentation quality, protein rumen degradability in situ, and organic matter digestibility in vitro through gas production after 120 d of conservation. Wilted alfalfa containing 0 and 4% tannin (T0 and T2) was harvested at 40% DM (wilting level I) and 53% DM (wilting level II) for bale (600 mm diameter) silage. Silages were analyzed for fermentation quality after 78 d of conservation. All the silages were well fermented with no butyric acid. Lab-scale silages showed reductions in ammonia, nonprotein nitrogen (NPN) and DM losses in T2 and T3 treatments, while the fermentation acid profiles were unaffected. In experiment 1, the untreated silage (T0) had the highest protein degradability after being incubated in the rumen. The addition of tannin reduced crude protein ruminal disappearance in a dose-dependent manner. However, the tannin reduced the organic matter digestibility by 5.1% for all of the tannin addition levels. The tannin positively affected the silage quality in the round bale silages, in particular reducing ammonia and NPN in the lowest wilting level. In both experiments, T2 treatment reduced proteolysis without any influence of DM on the binding reaction and reduced the NPN by 15% in comparison to the control.

  8. Milk fatty acid composition, rumen microbial population, and animal performances in response to diets rich in linoleic acid supplemented with chestnut or quebracho tannins in dairy ewes.

    PubMed

    Buccioni, A; Pauselli, M; Viti, C; Minieri, S; Pallara, G; Roscini, V; Rapaccini, S; Marinucci, M Trabalza; Lupi, P; Conte, G; Mele, M

    2015-02-01

    The aim of the study was to evaluate milk fatty acid (FA) profile, animal performance, and rumen microbial population in response to diets containing soybean oil supplemented or not with chestnut and quebracho tannins in dairy ewes. Eighteen Comisana ewes at 122±6 d in milking were allotted into 3 experimental groups. Diets were characterized by chopped grass hay administered ad libitum and by 800 g/head and day of 3 experimental concentrates containing 84.5 g of soybean oil/kg of dry matter (DM) and 52.8 g/kg of DM of bentonite (control diet), chestnut tannin extract (CHT diet), or quebracho tannin extract (QUE diet). The trial lasted 4 wk. Milk yield was recorded daily, and milk composition and blood parameters were analyzed weekly. At the end of the experiment, samples of rumen fluid were collected to analyze pH, volatile fatty acid profile, and the relative proportions of Butyrivibrio fibrisolvens and Butyrivibrio proteoclasticus in the rumen microbial population. Hepatic functionality, milk yield, and gross composition were not affected by tannin extracts, whereas milk FA composition was characterized by significant changes in the concentration of linoleic acid (CHT +2.77% and QUE +9.23%), vaccenic acid (CHT +7.07% and QUE +13.88%), rumenic acid (CHT -1.88% and QUE +24.24%), stearic acid (CHT + 8.71% and QUE -11.45%), and saturated fatty acids (CHT -0.47% and QUE -3.38%). These differences were probably due to the ability of condensed versus hydrolyzable tannins to interfere with rumen microbial metabolism, as indirectly confirmed by changes in the relative proportions of B. fibrisolvens and B. proteoclasticus populations and by changes in the molar proportions of volatile fatty acids. The effect of the CHT diet on the milk FA profile and microbial species considered in this trial was intermediate between that of QUE and the control diet, suggesting a differential effect of condensed and hydrolyzable tannins on rumen microbes. Compared with control animals

  9. Vaccination of cattle with a methanogen protein produces specific antibodies in the saliva which are stable in the rumen.

    PubMed

    Subharat, Supatsak; Shu, Dairu; Zheng, Tao; Buddle, Bryce M; Janssen, Peter H; Luo, Dongwen; Wedlock, D Neil

    2015-04-15

    Methane is produced in the rumen of cattle by a group of archaea (single-celled organisms forming a domain distinct from bacteria and eucarya) called methanogens. Vaccination against methanogens has the potential to reduce methane emissions by inducing antibodies in saliva which are transferred to the rumen and diminish the ability of methanogens to produce methane. Since it is likely that an effective vaccination strategy will need to produce high levels of methanogen-specific antibody in the saliva; the choice of adjuvant, route of vaccination and stability of saliva-derived antibody in the rumen all need to be considered. In this study, stability of IgA and IgG in rumen fluid was determined using an in vitro assay. IgA levels in cattle saliva were reduced by only 40% after 8h exposure to rumen contents while IgG levels were reduced by 80%. These results indicated that antibody is relatively stable in the bovine rumen. A trial was conducted in cattle to investigate induction of immune responses to a methanogen protein, recombinant glycosyl transferase protein (rGT2) from Methanobrevibacter ruminantium M1. Groups of cattle (n=6) were vaccinated subcutaneously with rGT2, formulated with Montanide ISA61 with or without the TLR4 agonist, monophosphoryl lipid A (MPL). A control group (n=6) was not vaccinated. Strong antigen-specific IgG and moderate IgA responses were measured in the serum and saliva of the vaccinated animals and antibody was also detected in the rumen.

  10. From rumen to industry

    PubMed Central

    2012-01-01

    The rumen is one of the most complicated and most fascinating microbial ecosystems in nature. A wide variety of microbial species, including bacteria, fungi and protozoa act together to bioconvert (ligno)cellulosic plant material into compounds, which can be taken up and metabolized by the ruminant. Thus, the rumen perfectly resembles a solution to a current industrial problem: the biorefinery, which aims at the bioconversion of lignocellulosic material into fuels and chemicals. We suggest to intensify the studies of the ruminal microbial ecosystem from an industrial microbiologists point of view in order to make use of this rich source of organisms and enzymes. PMID:22963386

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

    PubMed Central

    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

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

    PubMed

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

    2014-01-01

    Live yeast (Saccharomyces cerevisiae) constitutes an effective additive for animal production; its probiotic effect may be related to the concentrate-to-forage ratio (CTFR). The objective of this study was to assess the effects of S. cerevisiae (SC) on fiber degradation and rumen microbial populations in steers fed diets with different levels of dietary concentrate. Ten Simmental × Local crossbred steers (450 ± 50 kg BW) were assigned to a control group or an SC group. Both groups were fed the same basal diet but the SC group received SC supplementation (8 × 10(9) cfu/h/d through the ruminal fistula) following a two-period crossover design. Each period consisted of four phases, each of which lasted 17 d: 10 d for dietary adaptation, 6 d for degradation study, and 1 d for rumen sample collection. From the 1(st) to the 4(th) phase, steers were fed in a stepwise fashion with increasing CTFRs, i.e., 30:70, 50:50, 70:30, and 90:10. The kinetics of dry matter and fiber degradation of alfalfa pellets were evaluated; the rumen microbial populations were detected using real-time PCR. The results revealed no significant (P > 0.05) interactions between dietary CTFR and SC for most parameters. Dietary CTFR had a significant effect (P < 0.01) on degradation characteristics of alfalfa pellets and the copies of rumen microorganism; the increasing concentrate level resulted in linear, quadratic or cubic variation trend for these parameters. SC supplementation significantly (P < 0.05) affected dry matter (DM) and neutral detergent fiber (NDF) degradation rates (c DM, c NDF) and NDF effective degradability (EDNDF). Compared with the control group, there was an increasing trend of rumen fungi and protozoa in SC group (P < 0.1); copies of total bacteria in SC group were significantly higher (P < 0.05). Additionally, percentage of Ruminobacter amylophilus was significantly lower (P < 0.05) but percentage of Selenomonas ruminantium was

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

    PubMed Central

    2014-01-01

    Live yeast (Saccharomyces cerevisiae) constitutes an effective additive for animal production; its probiotic effect may be related to the concentrate-to-forage ratio (CTFR). The objective of this study was to assess the effects of S. cerevisiae (SC) on fiber degradation and rumen microbial populations in steers fed diets with different levels of dietary concentrate. Ten Simmental × Local crossbred steers (450 ± 50 kg BW) were assigned to a control group or an SC group. Both groups were fed the same basal diet but the SC group received SC supplementation (8 × 109 cfu/h/d through the ruminal fistula) following a two-period crossover design. Each period consisted of four phases, each of which lasted 17 d: 10 d for dietary adaptation, 6 d for degradation study, and 1 d for rumen sample collection. From the 1st to the 4th phase, steers were fed in a stepwise fashion with increasing CTFRs, i.e., 30:70, 50:50, 70:30, and 90:10. The kinetics of dry matter and fiber degradation of alfalfa pellets were evaluated; the rumen microbial populations were detected using real-time PCR. The results revealed no significant (P > 0.05) interactions between dietary CTFR and SC for most parameters. Dietary CTFR had a significant effect (P < 0.01) on degradation characteristics of alfalfa pellets and the copies of rumen microorganism; the increasing concentrate level resulted in linear, quadratic or cubic variation trend for these parameters. SC supplementation significantly (P < 0.05) affected dry matter (DM) and neutral detergent fiber (NDF) degradation rates (cDM, cNDF) and NDF effective degradability (EDNDF). Compared with the control group, there was an increasing trend of rumen fungi and protozoa in SC group (P < 0.1); copies of total bacteria in SC group were significantly higher (P < 0.05). Additionally, percentage of Ruminobacter amylophilus was significantly lower (P < 0.05) but percentage of Selenomonas ruminantium was significantly

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

    PubMed

    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

  15. Opportunities and challenges of applying recent advances in dairy cattle protein nutrition to beef cattle nutrition

    USDA-ARS?s Scientific Manuscript database

    Ruminant protein nutrition is greatly affected by the fermentation in the rumen because the microbial population in the rumen converts a majority of protein provided in the diet to microbial protein. This has both positive and negative consequences on ruminant protein nutrition. On one hand, the rum...

  16. Effect of casein infusion in the rumen, duodenum or both sites on factors affecting forage intake and performance of dairy cows fed red clover-grass silage.

    PubMed

    Khalili, H; Huhtanen, P

    2002-04-01

    Four lactating dairy cows were used in a balanced 4 x 4 Latin square design to examine the effects of casein infusion in the rumen, duodenum, or both on the intake of red clover-grass silage, chewing behavior, diet digestion, microbial protein synthesis, rumen fermentation, digestion, and passage kinetics, milk production, and milk composition. Duodenal infusion increased eating time, silage intake, rumen neutral detergent fiber pool, yields of milk, protein, and lactose, and concentration of milk protein, and tended to decrease that of milk fat. Ruminal infusion tended to decrease eating time, and increased significantly microbial protein synthesis, rumen ammonia-N concentration, molar proportions of isovalerate and valerate, digestion rate of digestible neutral detergent fiber, passage rate of indigestible neutral detergent fiber, and milk urea content, and decreased rumen neutral detergent fiber pool. Silage intake, and yields of milk, protein, and lactose were highest when casein was simultaneously infused in both sites.

  17. Increasing linseed supply in dairy cow diets based on hay or corn silage: Effect on enteric methane emission, rumen microbial fermentation, and digestion.

    PubMed

    Martin, C; Ferlay, A; Mosoni, P; Rochette, Y; Chilliard, Y; Doreau, M

    2016-05-01

    We investigated the effects of increasing extruded linseed supply in diets based on hay (H; experiment 1) or corn silage (CS; experiment 2) on enteric methane (CH4) emission, rumen microbial and fermentation parameters, and rumen and total-tract digestibility. In each experiment, 4 lactating Holstein cows fitted with cannulas at the rumen and proximal duodenum were used in a 4×4 Latin square design (28-d periods). Cows were fed ad libitum a diet [50:50 and 60:40 forage:concentrate on a dry matter (DM) basis for experiments 1 and 2, respectively] without supplementation (H0, CS0) or supplemented with extruded linseed at 5% (H5, CS5), 10% (H10, CS10), and 15% (H15, CS15) of dietary DM (i.e., 1.8, 3.6 and 5.4% total fatty acids added, respectively). All measurements were carried out during the last 8 d of each period. Linseed supply linearly decreased daily CH4 emission in cows fed H diets (from 486 to 289g/d for H0 to H15, on average) and CS diets (from 354 to 207g/d for CS0 to CS15, on average). The average decrease in CH4 per kilogram of DM intake was, respectively, -7, -15, and -38% for H5, H10, H15 compared with the H0 diet, and -4, -8, and -34% for CS5, CS10, and CS15 compared with the CS0 diet. The same dose-response effect was observed on CH4 emission in percent of gross energy intake, per kilogram of nutrient digested, and per kilogram of 4% fat- and 3.3% protein-corrected milk (FPCM) in both experiments. Changes in the composition of rumen volatile fatty acids in response to increasing linseed supply resulted in a moderate or marked linear decrease in acetate:propionate ratio for H or CS diets, respectively. The depressive effect of linseed on total protozoa concentration was linear for H diets (-15 to -40%, on average, for H5 to H15 compared with H0) and quadratic for CS diets (-17 to -83%, on average, for CS5 to CS15 compared with CS0). Concentration of methanogens was similar among H or CS diets. The energetic benefits from the decreased CH4 emission

  18. Evaluation of the National Research Council (2001) dairy model and derivation of new prediction equations. 2. Rumen degradable and undegradable protein.

    PubMed

    White, R R; Roman-Garcia, Y; Firkins, J L; Kononoff, P; VandeHaar, M J; Tran, H; McGill, T; Garnett, R; Hanigan, M D

    2017-05-01

    This work evaluated the National Research Council (NRC) dairy model (2001) predictions of rumen undegradable (RUP) and degradable (RDP) protein compared with measured postruminal non-ammonia, nonmicrobial (NANMN) and microbial N flows. Models were evaluated using the root mean squared prediction error (RMSPE) as a percent of the observed mean, mean and slope biases as percentages of mean squared prediction error (MSPE), and concordance correlation coefficient (CCC). The NRC (2001) over-estimated NANMN by 18% and under-estimated microbial N by 14%. Both responses had large mean biases (19% and 20% of MSPE, respectively), and NANMN had a slope bias (22% of MSPE). The NRC NANMN estimate had high RMSPE (46% of observed mean) and low CCC (0.37); updating feed library A, B, and C protein fractions and degradation rate (Kd) estimates with newer literature only marginally improved fit. The re-fit NRC models for NANMN and microbial N had CCC of 0.89 and 0.94, respectively. When compared with a prediction of NANMN as a static mean fraction of N intake, the re-derived NRC approach did not have improved fit. A protein system of intermediate complexity was derived in an attempt to estimate NANMN with improved fit compared with the static mean NANMN model. In this system, postruminal appearance of A, B, and C protein fractions were predicted in a feed-type specific manner rather than from estimated passage and degradation rates. In a comparison to independent data achieved through cross-validation, the new protein system improved RMSPE (34 vs. 36% of observed mean) and CCC (0.42 vs. 0.30) compared with the static mean NANMN model. When the NRC microbial N equation was re-derived, the RDP term dropped from the model. Consequently, 2 new microbial protein equations were formulated, both used a saturating (increasing at a decreasing rate) form: one saturated with respect to TDN and the other saturated over increasing intakes of rumen degraded starch and NDF. Both equations

  19. Effect of protein source and soluble carbohydrate addition on rumen fermentation and lactation performance of Holstein cows.

    PubMed

    McCormick, M E; Redfearn, D D; Ward, J D; Blouin, D C

    2001-07-01

    Rumen in vitro and in vivo experiments were conducted to evaluate the effect of rumen undegradable protein and soluble carbohydrates on rumen ammonia N release and lactation performance of Holstein cows. In the in vitro experiment, freeze-dried annual ryegrass (Lolium multiflorum, LAM) pasture was supplemented 1:1 with ground corn-based grain supplements containing expeller or solvent soybean meal with sucrose or lactose supplements at 0, 2.5, or 5% of dry matter (DM). The ammonia release rate was slower with expeller compared with solvent soybean meal-supplemented diets. Sucrose supplementation at the 5% level lowered rumen ammonia concentrations, but lactose-fortification of grain supplements was without effect. In the in vivo study, 32 multiparous Holstein cows were blocked according to milk yield and randomly assigned to corn-based grain supplements containing 1) solvent soybean meal, 2) solvent soybean meal + 5% sucrose supplement, 3) expeller soybean meal, or 4) expeller soybean meal + 5% sucrose supplement. Grain supplements and fresh annual ryegrass were component fed at approximately a 1:1 grain to forage ratio (DM basis). Forage DM intake was higher for cows receiving solvent soybean meal supplemented grain supplements than those receiving expeller soybean meal (12.2 +/- 2.1 vs. 11.4 +/- 2.2 kg/d), but total DM intake was similar for all diets (22.8 +/- 2.9 kg/d). Fat-corrected milk yield was similar for all diets averaging 37.5, 38.2, 39.1, and 37.6 kg/d for diets 1 to 4, respectively. Rumen fermentation, milk urea nitrogen, and body condition were unaffected by supplements; however, cows fed grain supplement 1 utilized dietary energy more efficiently than cows offered the other dietary treatments. High dietary crude protein concentrations may have limited lactation response to rumen undegradable protein and sugar.

  20. Copper Sulfate-induced Fermentation Changes in Continuous Cultures of the Rumen Microbial Ecosystem1

    PubMed Central

    Slyter, L. L.; Wolin, M. J.

    1967-01-01

    The effect of CuSO4 on fermentation was studied in a continuously cultured rumen ecosystem. CuSO4, introduced at a level of 50 mg/500 ml of culture volume twice daily, caused a marked inhibition of fermentation of concentrates. Fermentation of alfalfa hay was not inhibited by the same CuSO4 concentration when the inoculum for the culture was obtained from a cow maintained on a normal concentrate ration. When the inoculum was from a cow on a high concentrate ration, hay fermentation was partially inhibited by CuSO4. Concentrations of CuSO4 that did not inhibit the fermentation of alfalfa hay or hay-concentrate mixtures caused preferential production of propionic acid and decreased production of methane. PMID:6077413

  1. Effects of one-seed juniper and polyethylene glycol on intake, rumen fermentation, and plasma amino acids in sheep and goats fed supplemental protein and tannins.

    USDA-ARS?s Scientific Manuscript database

    We tested the effect of polyethylene glycol (PEG) on juniper and total intake, rumen fermentation, and plasma amino acids (AA) of 12 does and 12 ewes fed sudangrass and basal diets containing 10% quebracho tannins with no protein supplement (Control; 5% CP) or high rumen degradable (RDP 15% CP) or u...

  2. Diversity of phytases in the rumen.

    PubMed

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

    2007-01-01

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

  3. Estimating Rumen Degradable Protein in Forage Legume Hays and Silages by In Situ Disappearance Kinetics vs. Alternative Methods

    USDA-ARS?s Scientific Manuscript database

    Currently, the in situ kinetic method is considered the “gold standard” for estimating rumen degradable protein (RDP) in forages. When such estimates are impractical (e.g. numerous samples, limited sample quantities, or lack of ruminally fistulated cattle) alternative RDP methods are used, but their...

  4. Effects of extracts of Humulus lupulus (hops) and Yucca schidigera applied alone or in combination with monensin on rumen fermentation and microbial populations in vitro.

    PubMed

    Narvaez, Nelmy; Wang, Yuxi; McAllister, Tim

    2013-08-15

    β-Acids in hops (Humulus lupulus) and saponins in yucca (Yucca schidigera) have been found to possess antimicrobial properties similar to that of monensin and could be an alternative to in-feed antibiotics. The effects of monensin (MON) and ethanol extracts of hops (HE) and Y. schidigera (YE) alone and in combination with MON were assessed on ruminal microbial composition and fermentation in vitro of a barley-based diet. All treatments decreased (P < 0.05) CH4 production (per unit of dry matter), microbial protein (mg), and NH3 -N accumulation. All treatments reduced (P < 0.01) the acetate:propionate (A:P) ratio and molar proportions of butyrate, but increased (P < 0.01) those of propionate, whereas those of acetate decreased (P < 0.001) with addition of MON (10 µg mL(-1)) and combined with HE or YE. Methane produced per unit of true digested dry matter decreased (P < 0.001) with all treatments except YE. Monensin reduced (P < 0.001) proportions of 16S rRNA copies of Ruminococcus flavefaciens, but increased (P < 0.01) those of Selenomonas ruminantium. Hops extract alone or combined with MON reduced (P < 0.01) proportions of R. flavefaciens but combined with MON tended (P < 0.1) to increase those of S. ruminantium. Yucca extract combined with MON increased (P < 0.01) the proportions of R. flavefaciens and S. ruminantium. All treatments except MON (2.5 µg mL(-1)) reduced (P < 0.01) the relative abundance of methanogens. Hops extract and YE altered rumen microbes and fermentation in a manner similar to MON with many responses being additive when applied in combination. © 2013 Society of Chemical Industry.

  5. Estimation of feed crude protein concentration and rumen degradability by Fourier-transform infrared spectroscopy.

    PubMed

    Belanche, A; Weisbjerg, M R; Allison, G G; Newbold, C J; Moorby, J M

    2013-01-01

    Currently, rapid methods are needed for feed analysis. This study examined the potential of Fourier-transform infrared (FTIR) spectroscopy to predict the nutritional value of a wide range of feeds for ruminants, as an alternative to the in situ technique. Moreover, we investigated whether universal equations could be developed that would allow the low-cost determination of crude protein (CP) concentrations and their kinetics of degradation into the rumen. Protein nutritional values of 663 samples comprising 80 different feed types were determined in terms of concentrations of CP, water-soluble CP (CP(WS)), total-tract mobile bag CP digestibility (CP(TTD)), and in situ CP degradability, including the rumen soluble fraction (CP(A)), the degradable but not soluble fraction (CP(B)), rate of CP(B) degradation (CP(C)), effective degradability (CP(ED)), and potential degradability (CPPD). Infrared spectra of dry samples were collected by attenuated total reflectance from 4000 to 600 cm(-1). Models were developed by partial least squares (PLS) regression in a randomly selected subset of samples, and the precision of the equations was confirmed by using an external validation set. Analysis by FTIR spectroscopy was sufficiently sensitive to allow the accurate prediction of sample CP concentration (R(2)=0.92) and to classify feeds according to their CPWS concentrations using universal models (R(2)=0.78) that included all sample types. Moreover, substantial improvements in predictions were observed when samples were subdivided in groups. Models for forages led to accurate predictions of CP(WS) and fractions CP(A) and CP(B) (R(2)>0.83), whereas models for CP(TTD) and CP(ED) could be used for screening purposes (R(2)>0.67). This study showed that models for protein-rich concentrates alone could also be used for screening according to the feed concentrations of CP(WS), CP(TTD), CP(ED), CP(A), and CP(B), but models for energy-rich concentrates gave relatively poor predictions. The

  6. Direct electricity recovery from Canna indica by an air-cathode microbial fuel cell inoculated with rumen microorganisms.

    PubMed

    Zang, Guo-Long; Sheng, Guo-Ping; Tong, Zhong-Hua; Liu, Xian-Wei; Teng, Shao-Xiang; Li, Wen-Wei; Yu, Han-Qing

    2010-04-01

    Aquatic plants are widely used for phytoremediation, and effective disposal methods should be pursued for their utilization and to avoid further environmental pollution problems. This study demonstrated that, using an air-cathode microbial fuel cell (MFC) inoculated with rumen microorganisms, electricity could be directly produced with a maximum power density of 0.405 W/m(3) from Canna indica (canna), a lignocellulosic aquatic plant rich in cellulose, hemicellulose, and lignin, without pretreatment. The mechanisms of the Canna indica degradation in the MFC were elucidated through analyzing the changes of canna structure and intermediates, that is, soluble sugars and volatile fatty acids (VFAs), in the electricity generation process. The results showed that lignin was partially removed and more cellulose became exposed on the sample surface during the electricity generation in the MFC. The electron transfer in this MFC was mainly completed through electron shuttling via self-produced mediators. This work presents an attempt to understand how complex substrates like aquatic plants are decomposed in an MFC during electricity generation. It might, hopefully, provide a promising way to utilize lignocellulosic biomass for energy generation.

  7. Rumen microbial and fermentation characteristics are affected differently by bacterial probiotic supplementation during induced lactic and subacute acidosis in sheep

    PubMed Central

    2012-01-01

    Background Ruminal disbiosis induced by feeding is the cause of ruminal acidosis, a digestive disorder prevalent in high-producing ruminants. Because probiotic microorganisms can modulate the gastrointestinal microbiota, propionibacteria- and lactobacilli-based probiotics were tested for their effectiveness in preventing different forms of acidosis. Results Lactic acidosis, butyric and propionic subacute ruminal acidosis (SARA) were induced by feed chalenges in three groups of four wethers intraruminally dosed with wheat, corn or beet pulp. In each group, wethers were either not supplemented (C) or supplemented with Propionibacterium P63 alone (P) or combined with L. plantarum (Lp + P) or L. rhamnosus (Lr + P). Compared with C, all the probiotics stimulated lactobacilli proliferation, which reached up to 25% of total bacteria during wheat-induced lactic acidosis. This induced a large increase in lactate concentration, which decreased ruminal pH. During the corn-induced butyric SARA, Lp + P decreased Prevotella spp. proportion with a concomitant decrease in microbial amylase activity and total volatile fatty acids concentration, and an increase in xylanase activity and pH. Relative to the beet pulp-induced propionic SARA, P and Lr + P improved ruminal pH without affecting the microbial or fermentation characteristics. Regardless of acidosis type, denaturing gradient gel electrophoresis revealed that probiotic supplementations modified the bacterial community structure. Conclusion This work showed that the effectiveness of the bacterial probiotics tested depended on the acidosis type. Although these probiotics were ineffective in lactic acidosis because of a deeply disturbed rumen microbiota, some of the probiotics tested may be useful to minimize the occurrence of butyric and propionic SARA in sheep. However, their modes of action need to be further investigated. PMID:22812531

  8. Effects of variation of dietary sulfur on movement of sulfur in sheep rumen

    SciTech Connect

    Kandylis, K.; Bray, A.C.

    1987-01-01

    Effects of variations in dietary sulfur on rumen sulfur dynamics were studied under steady state conditions. In the first experimental period, three sheep were given 33.3 g of a pelleted diet hourly containing 1.59 g sulfur/kg (low) and in the second period the sulfur content was increased to 3.21 g/kg (high) by the addition of sodium sulfate. The daily sulfur intake was 1.158 g on the low sulfur diet and .545 g of this passed from the rumen in protein, .614 g was calculated to be absorbed from the rumen as sulfide, and .052 g was estimated to be recycled to the rumen. For sheep with daily intakes of 2.317 g sulfur, 1.212 g passed from the rumen in protein, 1.078 g was absorbed from the rumen, and .093 g was estimated to be recycled. It was estimated that 127 and 165 g microbial protein were synthesized/kg organic matter truly digested in the rumen for low and high sulfur diets, respectively. A simple model using simultaneous equations was proposed to describe rumen sulfur metabolism.

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

    PubMed

    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

  10. Biochemical and microbial analysis of ovine rumen fluid incubated with 1,3,5-trinitro-1,3,5-triazacyclohexane (RDX).

    PubMed

    Perumbakkam, Sudeep; Craig, A Morrie

    2012-08-01

    In this study, the rumen was assessed for its potential to detoxify RDX using molecular microbial ecology as well as analytical chemistry techniques. Results indicated significant loss (P < 0.05) of RDX in <8-h post incubation, and qualitative LC-MS/MS analysis showed evidence for the formation of 1-NO-RDX (M-O + HCOO) and methylenedinitramine metabolites. A total of 1106 16S rRNA-V3 clones were sequenced, and most sequences associated with either the phyla Bacteroidetes or Firmicutes. A LibCompare analysis for the RDX treatment showed an enrichment (P < 0.01) of the genus Prevotella. From these results, it can be concluded that the rumen is capable of detoxifying RDX, and the members of the genus Prevotella are linked to this detoxification.

  11. Differences in Ureolytic Bacterial Composition between the Rumen Digesta and Rumen Wall Based on ureC Gene Classification.

    PubMed

    Jin, Di; Zhao, Shengguo; Zheng, Nan; Bu, Dengpan; Beckers, Yves; Denman, Stuart E; McSweeney, Christopher S; Wang, Jiaqi

    2017-01-01

    Ureolytic bacteria are key organisms in the rumen producing urease enzymes to catalyze the breakdown of urea to ammonia for the synthesis of microbial protein. However, little is known about the diversity and distribution of rumen ureolytic microorganisms. The urease gene (ureC) has been the target gene of choice for analysis of the urea-degrading microorganisms in various environments. In this study, we investigated the predominant ureC genes of the ureolytic bacteria in the rumen of dairy cows using high-throughput sequencing. Six dairy cows with rumen fistulas were assigned to a two-period cross-over trial. A control group (n = 3) were fed a total mixed ration without urea and the treatment group (n = 3) were fed rations plus 180 g urea per cow per day at three separate times. Rumen bacterial samples from liquid and solid digesta and rumen wall fractions were collected for ureC gene amplification and sequencing using Miseq. The wall-adherent bacteria (WAB) had a distinct ureolytic bacterial profile compared to the solid-adherent bacteria (SAB) and liquid-associated bacteria (LAB) but more than 55% of the ureC sequences did not affiliate with any known taxonomically assigned urease genes. Diversity analysis of the ureC genes showed that the Shannon and Chao1 indices for the rumen WAB was lower than those observed for the SAB and LAB (P < 0.01). The most abundant ureC genes were affiliated with Methylococcaceae, Clostridiaceae, Paenibacillaceae, Helicobacteraceae, and Methylophilaceae families. Compared with the rumen LAB and SAB, relative abundance of the OTUs affiliated with Methylophilus and Marinobacter genera were significantly higher (P < 0.05) in the WAB. Supplementation with urea did not alter the composition of the detected ureolytic bacteria. This study has identified significant populations of ureolytic WAB representing genera that have not been recognized or studied previously in the rumen. The taxonomic classification of rumen ureC genes in the dairy

  12. Differences in Ureolytic Bacterial Composition between the Rumen Digesta and Rumen Wall Based on ureC Gene Classification

    PubMed Central

    Jin, Di; Zhao, Shengguo; Zheng, Nan; Bu, Dengpan; Beckers, Yves; Denman, Stuart E.; McSweeney, Christopher S.; Wang, Jiaqi

    2017-01-01

    Ureolytic bacteria are key organisms in the rumen producing urease enzymes to catalyze the breakdown of urea to ammonia for the synthesis of microbial protein. However, little is known about the diversity and distribution of rumen ureolytic microorganisms. The urease gene (ureC) has been the target gene of choice for analysis of the urea-degrading microorganisms in various environments. In this study, we investigated the predominant ureC genes of the ureolytic bacteria in the rumen of dairy cows using high-throughput sequencing. Six dairy cows with rumen fistulas were assigned to a two-period cross-over trial. A control group (n = 3) were fed a total mixed ration without urea and the treatment group (n = 3) were fed rations plus 180 g urea per cow per day at three separate times. Rumen bacterial samples from liquid and solid digesta and rumen wall fractions were collected for ureC gene amplification and sequencing using Miseq. The wall-adherent bacteria (WAB) had a distinct ureolytic bacterial profile compared to the solid-adherent bacteria (SAB) and liquid-associated bacteria (LAB) but more than 55% of the ureC sequences did not affiliate with any known taxonomically assigned urease genes. Diversity analysis of the ureC genes showed that the Shannon and Chao1 indices for the rumen WAB was lower than those observed for the SAB and LAB (P < 0.01). The most abundant ureC genes were affiliated with Methylococcaceae, Clostridiaceae, Paenibacillaceae, Helicobacteraceae, and Methylophilaceae families. Compared with the rumen LAB and SAB, relative abundance of the OTUs affiliated with Methylophilus and Marinobacter genera were significantly higher (P < 0.05) in the WAB. Supplementation with urea did not alter the composition of the detected ureolytic bacteria. This study has identified significant populations of ureolytic WAB representing genera that have not been recognized or studied previously in the rumen. The taxonomic classification of rumen ureC genes in the dairy

  13. Biochemical characterization of cellulose-binding proteins (CBPA and CBPB) from the rumen cellulolytic bacterium Eubacterium cellulosolvens 5.

    PubMed

    Yoshimatsu, Miho; Toyoda, Atsushi; Onizawa, Naoki; Nakamura, Yutaka; Minato, Hajime

    2007-10-01

    The cellulose-binding proteins, CBPA and CBPB, of rumen cellulolytic bacterium Eubacterium cellulosolvens 5 were biochemically characterized, and their properties were compared. Recombinant CBPA and CBPB were a typical 1,4-beta-endoglucanase. Both proteins bound to insoluble polysaccharides such as Avicel cellulose, acid swollen cellulose, lichenan, chitin, and oat spelt xylan. On the other hand, only recombinant CBPB bound to agarose and starch.

  14. Rumen dry matter and crude protein degradability of extracted or untreated oilseeds and Leucaena leucocephala leaves.

    PubMed

    Gralak, M A; Kamalu, T; von Keyserlingk, M A; Kulasek, G W

    1997-01-01

    A study was undertaken to determine the rumen DM and CP degradability characteristics of soyabean, canola seed, peanut, palm kernel and Leucaena leucocephala leaves. The oilseeds were either treated with n-hexane to extract the fat or left untreated. Nylon bags were incubated in each of four rumen cannulated sheep for 0, 2, 4, 6, 12, 24 and 48 h. Animals were fed on a diet consisting of meadow hay (ad libitum) and 150 g of concentrate twice daily. Fat extraction caused a decrease (P < or = 0.05) in DM disappearance of soyabean at 0, 2, 4, 6 and 12 h and of peanuts at all incubation times. CP disappearance from peanuts was reduced (P < or = 0.05) as a result of fat extraction at 0, 2, 4, 6 and 12 h. Fat extraction of canola seed increased CP disappearance at 0, 2, 4, 6 and 24 h (P < or = 0.05). However, in the case of defatted canola seed, an increase in DM disappearance (P < or = 0.05) was observed in the first 4 incubation times and a decrease (P < or = 0.05) in the later times. Fat extraction increased (P < or = 0.05) DM disappearance of palm kernel at 0 and 48 h, but reduced it at 4, 6 and 24 h. CP disappearance of palm kernel was improved by treatment (P < or = 0.05) at 0, 4, 24 and 48 h and decreased at 12 h. In the case of palm kernel the largest differences in DM and CP disappearance occurred between the 24 and 48 h incubation times. Degradability characteristics for DM and CP of full-fat soyabean, canola seed and peanut were comparable to those of the full fat samples. Effective DM degradability of soyabean, canola seed and peanuts was 72.2 and 71.9; 74.1 and 66.8; and 85.9 and 70.8 for full fat and extracted feeds, respectively. Effective CP degradability was similar in all oilseeds with the exception of the extracted canola seed. Therefore, the incorporation of full-fat soyabean, canola seed and peanut into ruminant rations can be considered as a means of increasing the energy balance. Both palm kernel DM and CP degradabilities were characterized by slow

  15. Comparative Survey of Rumen Microbial Communities and Metabolites across One Caprine and Three Bovine Groups, Using Bar-Coded Pyrosequencing and 1H Nuclear Magnetic Resonance Spectroscopy

    PubMed Central

    Lee, Hyo Jung; Jung, Ji Young; Oh, Young Kyoon; Lee, Sang-Suk; Madsen, Eugene L.

    2012-01-01

    Pyrosequencing of 16S rRNA genes (targeting Bacteria and Archaea) and 1H nuclear magnetic resonance were applied to investigate the rumen microbiota and metabolites of Hanwoo steers in the growth stage (HGS), Hanwoo steers in the late fattening stage (HFS), Holstein-Friesian dairy cattle (HDC), and Korean native goats (KNG) in the late fattening stage. This was a two-part investigation. We began by comparing metabolites and microbiota of Hanwoo steers at two stages of husbandry. Statistical comparisons of metabolites and microbial communities showed no significant differences between HFS and HGS (differing by a dietary shift at 24 months and age [67 months versus 12 months]). We then augmented the study by extending the investigation to HDC and KNG. Overall, pyrosequencing of 16S rRNA genes showed that the rumens had highly diverse microbial communities containing many previously undescribed microorganisms. Bioinformatic analysis revealed that the bacterial sequences were predominantly affiliated with four phyla—Bacteroidetes, Firmicutes, Fibrobacteres, and Proteobacteria—in all ruminants. However, interestingly, the bacterial reads belonging to Fibrobacteres were present at a very low abundance (<0.1%) in KNG. Archaeal community analysis showed that almost all of these reads fell into a clade related to, but distinct from, known cultivated methanogens. Statistical analyses showed that the microbial communities and metabolites of KNG were clearly distinct from those of other ruminants. In addition, bacterial communities and metabolite profiles of HGS and HDC, fed similar diets, were distinctive. Our data indicate that bovine host breeds override diet as the key factor that determines bacterial community and metabolite profiles in the rumen. PMID:22706048

  16. Effects of essential oils on digestion, ruminal fermentation, rumen microbial populations, milk production, and milk composition in dairy cows fed alfalfa silage or corn silage.

    PubMed

    Benchaar, C; Petit, H V; Berthiaume, R; Ouellet, D R; Chiquette, J; Chouinard, P Y

    2007-02-01

    Four Holstein cows fitted with ruminal cannulas were used in a 4 x 4 Latin square design (28-d periods) with a 2 x 2 factorial arrangement of treatments to investigate the effects of addition of a specific mixture of essential oil compounds (MEO; 0 vs. 750 mg/d) and silage source [alfalfa silage (AS) vs. corn silage (CS)] on digestion, ruminal fermentation, rumen microbial populations, milk production, and milk composition. Total mixed rations containing either AS or CS as the sole forage source were balanced to be isocaloric and isonitrogenous. In general, no interactions between MEO addition and silage source were observed. Except for ruminal pH and milk lactose content, which were increased by MEO supplementation, no changes attributable to the administration of MEO were observed for feed intake, nutrient digestibility, end-products of ruminal fermentation, microbial counts, and milk performance. Dry matter intake and milk production were not affected by replacing AS with CS in the diet. However, cows fed CS-based diets produced milk with lower fat and higher protein and urea N concentrations than cows fed AS-based diets. Replacing AS with CS increased the concentration of NH(3)-N and reduced the acetate-to-propionate ratio in ruminal fluid. Total viable bacteria, cellulolytic bacteria, and protozoa were not influenced by MEO supplementation, but the total viable bacteria count was higher with CS- than with AS-based diets. The apparent digestibility of crude protein did not differ between the AS and CS treatments, but digestibilities of neutral detergent fiber and acid detergent fiber were lower when cows were fed CS-based diets than when they were fed AS-based diets. Duodenal bacterial N flow, estimated using urinary purine derivatives and the amount of N retained, increased in cows fed CS-based diets compared with those fed AS-based diets. Feeding cows AS increased the milk fat contents of cis-9, trans-11 18:2 (conjugated linoleic acid) and 18:3 (n-3 fatty

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

    PubMed Central

    Norrapoke, T.; Wanapat, M.; Wanapat, S.

    2012-01-01

    Four, lactating dairy crossbreds (50%×50% Holstein Friesian×Native Zebu cattle) were randomly assigned according to a 2×2 factorial arrangement (two protein levels and two levels of mangosteen peel pellets (Mago-pel)) in a 4×4 Latin square design to receive four dietary treatments. All cows received concentrate at a proportion of 1 kg concentrate per 2 kg of milk yield, and urea-treated 5% rice straw (UTRS) was given ad libitum. It was found that total dry matter intakes, nutrient digestibility, ruminal pH and NH3-N concentrations were not affected (p>0.05) by treatments. Concentrations of ruminal pH and NH3-N were not affected by dietary treatments although the concentration of BUN varied significantly (p<0.05) between protein levels (p<0.05). The populations of rumen bacteria and fungal zoospores did not differ among treatments (p>0.05); however, the population of protozoa was decreased (p<0.05) when cows received Mago-pel supplementation. The composition of the population of bacteria, identified by real-time PCR technique, including total bacteria, methanogens, Fibrobacter succinogenes and Ruminococcus albus was similar (p>0.05) among dietary treatments (p>0.05); however, copy numbers of Ruminococcus flavefaciens was increased when protein level increased (p<0.05). Microbial protein synthesis, in terms of both quantity and efficiency, was enriched by Mago-pel supplementation. Milk yield was greatest in cows fed UTRS based diets with concentrate containing protein at 16% CP with Mago-pel, but were lowest without Mago-pel (p<0.05). In addition, protein level and supplementation of Mago-pel did not affect (p>0.05) milk composition except solids-not-fat which was higher in cows fed the diet with 19% CP. Therefore, feeding a concentrate containing 16% CP together with 300 g/hd/d Mago-pel supplementation results in changes in rumen fermentation and microbial population and improvements in milk production in lactating dairy crossbreds fed on UTRS. PMID:25049652

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

    PubMed

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

    2009-08-01

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

  19. Effect of the combination of crude extracts of Penicillium griseofulvum and Fusarium graminearum containing patulin and zearalenone on rumen microbial fermentation and on their metabolism in continuous culture fermenters.

    PubMed

    Riccio, María Belén; Tapia, María Ofelia; Martínez, Guadalupe; Aranguren, Sandra Mariela; Dieguez, Susana Neyi; Soraci, Alejandro Luis; Rodríguez, Edgardo

    2014-01-01

    Six single-flow continuous cultures were used to study the effects of the mycotoxins patulin (PAT) and zearalenone (ZEN) alone or in combination on rumen microbial fermentation. In each of the four 7-d periods, the fermenters were supplemented in a 2 × 3 factorial arrangement with two levels of PAT (0 and 20 mg/l) and three levels of ZEN (0, 5 and 10 mg/l). The treatments did not affect the apparent and true digestibility of organic matter. PAT alone decreased the digestibility of neutral detergent fibre (NDF) and acid detergent fibre (ADF) (p < 0.01), but in the presence of 5 or 10 mg/l of ZEN, there were no effects of PAT. In contrast, the digestibility of NDF and ADF was decreased at 10 mg/l of ZEN in the absence of PAT (p < 0.05). The pH of the fermenters increased after 2 and 3 d of PAT treatment (p < 0.01). PAT decreased the concentration of total volatile acids (VFA), the molar proportion of acetate and the acetate:proportionate ratio (p < 0.01). The molar concentrations of other VFA were unchanged. Ammonia N (NH3-N) flow increased (p < 0.05) and there was a tendency to a higher NH3-N concentration (p < 0.1) in fermenters with PAT. Total N, non-ammonia N and bacterial N as well as efficiency of microbial protein synthesis and efficiency of N utilisation were not affected by treatments. PAT was nearly completely degraded during incubation. The mean recovery of ZEN, α-zearalenol and β-zearalenol expressed as a proportion of administered ZEN was less than 50% in effluents from fermenters receiving only ZEN and ZEN plus PAT, respectively. With exception of fibre digestion, the co-administration of PAT and ZEN did not elicit interaction effects on most measured parameters of rumen metabolism.

  20. Microbial β-glucosidases from cow rumen metagenome enhance the saccharification of lignocellulose in combination with commercial cellulase cocktail

    PubMed Central

    2012-01-01

    Background A complete saccharification of plant polymers is the critical step in the efficient production of bio-alcohols. Beta-glucosidases acting in the degradation of intermediate gluco-oligosaccharides produced by cellulases limit the yield of the final product. Results In the present work, we have identified and then successfully cloned, expressed, purified and characterised 4 highly active beta-glucosidases from fibre-adherent microbial community from the cow rumen. The enzymes were most active at temperatures 45–55°C and pH 4.0-7.0 and exhibited high affinity and activity towards synthetic substrates such as p-nitrophenyl-beta-D-glucopyranoside (pNPbetaG) and pNP-beta-cellobiose, as well as to natural cello-oligosaccharides ranging from cellobiose to cellopentaose. The apparent capability of the most active beta-glucosidase, herein named LAB25g2, was tested for its ability to improve, at low dosage (31.25 units g-1 dry biomass, using pNPbetaG as substrate), the hydrolysis of pre-treated corn stover (dry matter content of 20%; 350 g glucan kg-1 dry biomass) in combination with a beta-glucosidase-deficient commercial Trichoderma reseei cellulase cocktail (5 units g-1 dry biomass in the basis of pNPbetaG). LAB25g2 increased the final hydrolysis yield by a factor of 20% (44.5 ± 1.7% vs. 34.5 ± 1.5% in control conditions) after 96–120 h as compared to control reactions in its absence or in the presence of other commercial beta-glucosidase preparations. The high stability (half-life higher than 5 days at 50°C and pH 5.2) and 2–38000 fold higher (as compared with reported beta-glucosidases) activity towards cello-oligosaccharides may account for its performance in supplementation assays. Conclusions The results suggest that beta-glucosidases from yet uncultured bacteria from animal digestomes may be of a potential interest for biotechnological processes related to the effective bio-ethanol production in combination with low dosage of commercial cellulases

  1. Tuning microbial hosts for membrane protein production

    PubMed Central

    2009-01-01

    The last four years have brought exciting progress in membrane protein research. Finally those many efforts that have been put into expression of eukaryotic membrane proteins are coming to fruition and enable to solve an ever-growing number of high resolution structures. In the past, many skilful optimization steps were required to achieve sufficient expression of functional membrane proteins. Optimization was performed individually for every membrane protein, but provided insight about commonly encountered bottlenecks and, more importantly, general guidelines how to alleviate cellular limitations during microbial membrane protein expression. Lately, system-wide analyses are emerging as powerful means to decipher cellular bottlenecks during heterologous protein production and their use in microbial membrane protein expression has grown in popularity during the past months. This review covers the most prominent solutions and pitfalls in expression of eukaryotic membrane proteins using microbial hosts (prokaryotes, yeasts), highlights skilful applications of our basic understanding to improve membrane protein production. Omics technologies provide new concepts to engineer microbial hosts for membrane protein production. PMID:20040113

  2. Rumen Metagenomics

    USDA-ARS?s Scientific Manuscript database

    The rumen microbiome plays a critical role in normal physiology and nutrition of ruminants. Alterations in the rumen microbiome have important physiological and pathological implications. The advent of next-generation sequencing technologies and rapid development of computational tools and reference...

  3. Supplementation of cattle fed tropical grasses with microalgae increases microbial protein production and average daily gain.

    PubMed

    Costa, D F A; Quigley, S P; Isherwood, P; McLennan, S R; Poppi, D P

    2016-05-01

    A series of 3 experiments were conducted to evaluate the use of microalgae as supplements for ruminants consuming low-CP tropical grasses. In Exp. 1, the chemical composition and in vitro protein degradability of 9 algae species and 4 protein supplements were determined. In Exp. 2, rumen function and microbial protein (MCP) production were determined in steers fed speargrass hay alone or supplemented with , , , or cottonseed meal (CSM). In Exp. 3, DMI and ADG were determined in steers fed speargrass hay alone or supplemented with increasing amounts of NPN (urea combined with ammonia sulfate), CSM, or . In Exp. 1, the CP content of and (675 and 580 g/kg DM) was highest among the algae species and higher than the other protein supplements evaluated, and sp. had the highest crude lipid (CL) content (198 g/kg DM). In Exp. 2, supplementation increased speargrass hay intake, the efficiency of MCP production, the fractional outflow rate of digesta from the rumen, the concentration of NHN, and the molar proportion of branched-chain fatty acids in the rumen fluid of steers above all other treatments. acceptance by steers was low and this resulted in no significant difference to unsupplemented steers for all parameters measured for this algae supplement. In Exp. 3, ADG linearly increased with increasing supplementary N intake from both and NPN, with no difference between the 2 supplements. In contrast, ADG quadratically increased with increasing supplementary N intake from CSM. It was concluded that and may potentially be used as protein sources for cattle grazing low-CP pastures.

  4. Rumen degradability characteristics of normal maize stover and silage, and quality protein maize silage-based diets offered to cows.

    PubMed

    Tamir, Berhan; Gebrehawariat, Ephrem; Tegegne, Azage; Kortu, Mohammed Y

    2012-10-01

    Rumen degradability characteristics of dry matter (DM), organic matter (OM) and crude protein (CP) of normal maize (NM) stover (T1)-, NM silage (T2)- and quality protein maize (QPM) silage (T3)-based diets were studied using three rumen-fistulated Boran × Friesian non-lactating cows (371 ± 32.00 kg) in 3 × 3 Latin Square Design. Cows were supplemented with a similar concentrate mix. In sacco degradability of DM and OM indicated that the (a) values of DM (128) and OM (114) for NM stover were lower (P < 0.001) than that for NM silage (268 and 253) and for QPM silage (323 and 303), respectively. The (a) value for CP was lower (P < 0.05) for QPM silage (286) than for NM stover (404) and NM silage (326). The (b) values of DM in NM stover (597) and NM silage (535) were higher (P < 0.05) than in QPM silage (499). The (b) value of CP in NM stover (372) was lower (P < 0.05) than in NM silage (655) and in QPM silage (608). Rate of degradation of OM in NM stover and NM silage, each with 0.03, was faster (P < 0.01) than in QPM silage (0.02). Moreover, QPM silage had higher potentially degradable fraction for DM (821) (P < 0.05) and OM (840) (P < 0.01) than DM (725) and OM (712) in NM stover. The mean rumen ammonia concentration (209 mg/l) of QPM silage was higher (P < 0.05) than that of NM stover (179 mg/l) and NM silage (170 mg/l). The average rumen pH (6.1) in cows fed QPM silage was lowest (P < 0.05) compared to pH (6.3) in cows fed either NM stover or silage. The concentration of total volatile fatty acids (116 mmol/l) in the rumen of cows incubated with QPM silage was higher (P < 0.001) than in those incubated with NM stover (113 mmol/l) and NM silage (110 mmol/l). It was concluded that QPM silage-based diet was superior in DM and OM degradability, and had higher ammonia and VFA concentration than NM stover-based diet. No differences have been observed in all parameters measured between QPM and NM silages.

  5. Changes of Microbial Population in the Rumen of Dairy Steers as Influenced by Plant Containing Tannins and Saponins and Roughage to Concentrate Ratio

    PubMed Central

    Anantasook, N.; Wanapat, M.; Cherdthong, A.; Gunun, P.

    2013-01-01

    The objective of this study was to investigate microbial population in the rumen of dairy steers as influenced by supplementing with dietary condensed tannins and saponins and different roughage to concentrate ratios. Four, rumen fistulated dairy steers (Bos indicus) were used in a 2×2 factorial arrangement in a 4×4 Latin square design. The main factors were two roughage to concentrate ratios (R:C, 60:40 and 40:60) and two supplementations of rain tree pod meal (RPM) (0 and 60 g/kg of total DM intake). Chopped 30 g/kg urea treated rice straw was used as a roughage source. All animals received feed according to respective R:C ratios at 25 g/kg body weight. The RPM contained crude tannins and saponins at 84 and 143 g/kg of DM, respectively. It was found that ruminal pH decreased while ruminal temperature increased by a higher concentrate ratio (R:C 40:60) (p<0.05). In contrast, total bacterial, Ruminococus albus and viable proteolytic bacteria were not affected by dietary supplementation. Numbers of fungi, cellulolytic bacteria, Fibrobactor succinogenes and Ruminococus flavefaciens were higher while amylolytic bacteria was lower when steers were fed at 400 g/kg of concentrate. The population of Fibrobactor succinogenes, was found to be higher with RPM supplementation. In addition, the use of real-time PCR technique indicated that the population of protozoa and methanogens were decreased (p<0.05) with supplementation of RPM and with an increasing concentrate ratio. Supplementation of RPM and feeding different concentrate ratios resulted in changing the rumen microbes especially, when the animals were fed at 600 g/kg of concentrate and supplemented with RPM which significantly reduced the protozoa and methanogens population. PMID:25049745

  6. Effects of Adaptation of In vitro Rumen Culture to Garlic Oil, Nitrate, and Saponin and Their Combinations on Methanogenesis, Fermentation, and Abundances and Diversity of Microbial Populations.

    PubMed

    Patra, Amlan K; Yu, Zhongtang

    2015-01-01

    This study investigated the effects of garlic oil (0.25 g/L), nitrate (5 mM), and quillaja saponin (0.6 g/L), alone and in binary or ternary combinations, on methanogenesis, rumen fermentation, and abundances of select microbial populations using in vitro rumen cultures. Potential adaptation to these compounds was also examined by repeated transfers of the cultures on alternate days until day 18. All treatments except saponin alone significantly decreased methanogenesis. Ternary combinations of garlic oil, nitrate, and saponin additively/synergistically suppressed methane production by 65% at day 2 and by 40% at day 18. Feed digestion was not adversely affected by any of the treatments at day 2, but was decreased by the combinations (binary and ternary) of garlic oil with the other inhibitors at days 10 and 18. Saponin, alone or in combinations, and garlic oil alone lowered ammonia concentration at day 2, while nitrate increased ammonia concentration at days 10 and 18. Total volatile fatty acid concentration was decreased by garlic oil alone or garlic oil-saponin combination. Molar proportions of acetate and propionate were affected to different extents by the different treatments. The abundances of methanogens were similar among treatments at day 2; however, garlic oil and its combination with saponin and/or nitrate at day 10 and all treatments except saponin at day 18 significantly decreased the abundances of methanogens. All the inhibitors, either alone or in combinations, did not adversely affect the abundances of total bacteria or Ruminococcus flavefaciens. However, at day 18 the abundances of Fibrobacter succinogenes and Ruminococcus albus were lowered in the presence of garlic oil and saponin, respectively. The results suggest that garlic oil-nitrate-saponin combination (at the doses used in this study) can effectively decreases methanogenesis in the rumen, but its efficacy may decrease while inhibition to feed digestion can increase over time.

  7. Effects of Adaptation of In vitro Rumen Culture to Garlic Oil, Nitrate, and Saponin and Their Combinations on Methanogenesis, Fermentation, and Abundances and Diversity of Microbial Populations

    PubMed Central

    Patra, Amlan K.; Yu, Zhongtang

    2015-01-01

    This study investigated the effects of garlic oil (0.25 g/L), nitrate (5 mM), and quillaja saponin (0.6 g/L), alone and in binary or ternary combinations, on methanogenesis, rumen fermentation, and abundances of select microbial populations using in vitro rumen cultures. Potential adaptation to these compounds was also examined by repeated transfers of the cultures on alternate days until day 18. All treatments except saponin alone significantly decreased methanogenesis. Ternary combinations of garlic oil, nitrate, and saponin additively/synergistically suppressed methane production by 65% at day 2 and by 40% at day 18. Feed digestion was not adversely affected by any of the treatments at day 2, but was decreased by the combinations (binary and ternary) of garlic oil with the other inhibitors at days 10 and 18. Saponin, alone or in combinations, and garlic oil alone lowered ammonia concentration at day 2, while nitrate increased ammonia concentration at days 10 and 18. Total volatile fatty acid concentration was decreased by garlic oil alone or garlic oil-saponin combination. Molar proportions of acetate and propionate were affected to different extents by the different treatments. The abundances of methanogens were similar among treatments at day 2; however, garlic oil and its combination with saponin and/or nitrate at day 10 and all treatments except saponin at day 18 significantly decreased the abundances of methanogens. All the inhibitors, either alone or in combinations, did not adversely affect the abundances of total bacteria or Ruminococcus flavefaciens. However, at day 18 the abundances of Fibrobacter succinogenes and Ruminococcus albus were lowered in the presence of garlic oil and saponin, respectively. The results suggest that garlic oil-nitrate-saponin combination (at the doses used in this study) can effectively decreases methanogenesis in the rumen, but its efficacy may decrease while inhibition to feed digestion can increase over time. PMID:26733975

  8. Effects of replacing dietary starch with neutral detergent-soluble fibre on ruminal fermentation, microbial synthesis and populations of ruminal cellulolytic bacteria using the rumen simulation technique (RUSITEC).

    PubMed

    Zhao, X H; Liu, C J; Liu, Y; Li, C Y; Yao, J H

    2013-12-01

    A rumen simulation technique (RUSITEC) apparatus with eight 800 ml fermenters was used to investigate the effects of replacing dietary starch with neutral detergent-soluble fibre (NDSF) by inclusion of sugar beet pulp in diets on ruminal fermentation, microbial synthesis and populations of ruminal cellulolytic bacteria. Experimental diets contained 12.7, 16.4, 20.1 or 23.8% NDSF substituted for starch on a dry matter basis. The experiment was conducted over two independent 15-day incubation periods with the last 8 days used for data collection. There was a tendency that 16.4% NDSF in the diet increased the apparent disappearance of organic matter (OM) and neutral detergent fibre (NDF). Increasing dietary NDSF level increased carboxymethylcellulase and xylanase activity in the solid fraction and apparent disappearance of acid detergent fibre (ADF) but reduced the 16S rDNA copy numbers of Ruminococcus albus in both liquid and solid fractions and R. flavefaciens in the solid fraction. The apparent disappearance of dietary nitrogen (N) was reduced by 29.6% with increased dietary NDSF. Substituting NDSF for starch appeared to increase the ratios of acetate/propionate and methane/volatile fatty acids (VFA) (mol/mol). Replacing dietary starch with NDSF reduced the daily production of ammonia-N and increased the growth of the solid-associated microbial pellets (SAM). Total microbial N flow and efficiency of microbial synthesis (EMS), expressed as g microbial N/kg OM fermented, tended to increase with increased dietary NDSF, but the numerical increase did not continue as dietary NDSF exceeded 20.1% of diet DM. Results suggested that substituting NDSF for starch up to 16.4% of diet DM increased digestion of nutrients (except for N) and microbial synthesis, and further increases (from 16.4% to 23.8%) in dietary NDSF did not repress microbial synthesis but did significantly reduce digestion of dietary N.

  9. A comparison of methods for the estimation of the proportion of microbial nitrogen in duodenal digesta, and of correction for microbial contamination in nylon bags incubated in the rumen of sheep.

    PubMed

    Kennedy, P M; Hazlewood, G P; Milligan, L P

    1984-09-01

    Four sheep, each fitted with cannulas in the rumen and proximal duodenum, were given two diets (1390 g dry matter (DM)/d) consisting of lucerne (Medicago sativa) pellets (24.2 g nitrogen/kg DM) plus pelleted reed canary grass (Phalaris arundinacea; 14.1 g N/kg DM) or chopped hay (11.8 g N/kg DM) at intervals of 2 h. Flow of duodenal digesta measured by reference to the markers 51Cr-EDTA and 103Ru-phenanthroline indicated a net gain of 5.8-7.5 g non-ammonia-N (NAN) between mouth and duodenum. The proportion of microbial N in duodenal digesta N was estimated using 15N and 35S incorporation into bacteria and digesta. Two methods of analysis for 35S content, the Bird & Fountain (1970; B&F method) and the Mathers & Miller (1980; M&M method), were used. (15NH4)2SO4 and Na2(35)SO4 were infused into the rumen for 3.5 d before and 4.0 d during sampling. A bacterial fraction was prepared from the fluid phases of sampled duodenal digesta and rumen contents by differential centrifugation. In addition, samples of ground canary grass and of lucerne were incubated in nylon bags in the rumen for 3-48 h during the infusion. Each of the 35S analytical methods yielded similar values of 35S content of isolated rumen or duodenal bacteria, but there was more (P less than 0.05) incorporation of 15N into rumen than into duodenal bacteria. Relative to values obtained using the M&M method and 15N incorporation, the B&F method for S analysis yielded higher (P less than 0.05) estimates of microbial content of duodenal digesta from sheep given chopped reed canary grass. 35S activity associated with washed nylon-bag residues increased rapidly with time-period of incubation and was substantially greater (P less than 0.05) when analysed by the B&F method compared with the M&M method. The 35S content (/g DM) of adherent bacteria removed from nylon-bag residues by homogenization in a second experiment varied from 0.65 to 1.88 that of free-living bacteria isolated from rumen fluid by differential

  10. The large bowel--a supplementary rumen?

    PubMed

    Argenzio, R A; Stevens, C E

    1984-01-01

    The rumen and the mammalian large intestine are similar in many respects. Microbial protein appears to be synthesized and degraded in the digesta of both organs in a comparable manner. The VFA end-products of carbohydrate fermentation are produced in similar concentrations. Digesta pH is maintained with buffer added by the saliva or ileal fluid, HCO3 released into the lumen and rapid absorption of the organic acids. VFA are absorbed at equivalent rates by rumen epithelium and large intestinal mucosa. Over-production of VFA produces similar adverse effects. There is a considerable amount of species variation in the relative length and volume as well as the extent of sacculation of the large intestine. The caecum is the primary site for retention of digesta and microbial fermentation in the large intestine of rabbits, rodents and a few other species. However, the proximal colon is the major site of retention and fermentation in most mammals. Absorptions of Na and VFA appear to account for absorption of most of the water removed during passage of digesta through the large intestine. A relatively slow rate of Na absorption and release of HCO3 appears to provide the fluid and buffering capacity needed for efficient microbial digestion in the rumen and in the large intestine of some species. A more rapid absorption of Na by the large intestine of other species would aid in the conservation of Na and water. The many similarities between the large intestine and the rumen suggest that further comparison can provide additional information on both the function and diseases of these two organs. The rumen has proved to be accessible to a variety of procedures useful for the study of microbial digestive processes and its epithelium has provided a non-glandular tissue for studies of inorganic ion transport as well as the transport and metabolism of VFA. Comparative studies of the large intestine also can provide a better understanding of the functions and malfunctions of the

  11. Comparison of fermentation of diets of variable composition and microbial populations in the rumen of sheep and Rusitec fermenters. II. Protozoa population and diversity of bacterial communities.

    PubMed

    Martínez, M E; Ranilla, M J; Tejido, M L; Saro, C; Carro, M D

    2010-08-01

    Four ruminally and duodenally cannulated sheep and 8 Rusitec fermenters were used to determine the effects of dietary characteristics on microbial populations and bacterial diversity. The purpose of the study was to assess how closely fermenters can mimic the differences between diets found in vivo. The 4 experimental diets contained forage to concentrate (F:C) ratios of 70:30 (high forage; HF) or 30:70 (high concentrate; HC) with either alfalfa hay (A) or grass hay (G) as the forage. Total bacterial numbers were greater in the rumen of sheep fed HF diets compared with those fed HC diets, whereas the opposite was found in fermenters. The numbers of cellulolytic bacteria were not affected by F:C ratio in any fermentation system, but cellulolytic numbers were 2.7 and 1.8 times greater in sheep than in fermenters for HF and HC diets, respectively. Neither total bacterial nor cellulolytic numbers were affected by the type of forage in sheep or fermenters. Decreasing F:C ratio increased total protozoa and Entodiniae numbers in sheep by about 29 and 25%, respectively, but it had no effect in fermenters. Isotrichidae and Ophryoscolecinae numbers in sheep were not affected by changing F:C ratio, but both disappeared completely from fermenters fed HC diets. Total protozoa and Entodiniae numbers were greater in sheep fed A diets than in those fed G diets, whereas the opposite was found in fermenters. Results indicate that under the conditions of the present study, protozoa population in Rusitec fermenters was not representative of that in the rumen of sheep fed the same diets. In addition, protozoa numbers in fermenters were 121 and 226 times lower than those in the sheep rumen for HF and HC diets, respectively. The automated ribosomal intergenic spacer analysis of the 16S ribosomal DNA was used to analyze the diversity of liquid- and solid-associated bacteria in both systems. A total of 170 peaks were detected in the automated ribosomal intergenic spacer analysis

  12. Biohydrogenation of dietary n-3 PUFA and stability of ingested vitamin E in the rumen, and their effects on microbial activity in sheep.

    PubMed

    Chikunya, S; Demirel, G; Enser, M; Wood, J D; Wilkinson, R G; Sinclair, L A

    2004-04-01

    The present study investigated the susceptibility of dietary n-3 PUFA to ruminal biohydrogenation, the stability of ingested vitamin E in the rumen and the subsequent uptake of PUFA and vitamin E into plasma. Six cannulated sheep were assigned to six diets over five 33 d periods, in an incomplete 6x5 Latin square. The diets, based on dried grass, were formulated to supply 50 g fatty acids/kg DM using three lipid sources: Megalac (calcium soap of palm fatty acid distillate; Volac Ltd, Royston, Herts., UK), linseed (formaldehyde-treated; Trouw Nutrition, Northwich, Ches., UK) and linseed-fish oil (formaldehyde-treated linseed+fish oil). The diets were supplemented with 100 or 500 mg alpha-tocopheryl acetate/kg DM. Fat source or level of vitamin E in the diet did not alter microbial activity in the rumen. Biohydrogenation of linoleic acid (18 : 3n-6; 85-90 %), linolenic acid (18 : 3n-3; 88-93 %), docosahexaenoic acid (22 : 6n-3; 91 %) and EPA (20 : 5n-3; 92 %) was extensive. Feeding formaldehyde-treated linseed elevated concentrations of 18 : 3n-3 in plasma, whilst 22 : 6n-3 and 20 : 5n-3 were only increased by feeding the linseed-fish oil blend. Duodenal recovery of ingested vitamin E was high (range 0.79-0.92 mg/mg fed). High dietary vitamin E was associated with increased plasma alpha-tocopherol (2.57 v. 1.46 microg/ml for 500 and 100 mg alpha-tocopheryl acetate/kg DM respectively), although all concentrations were low. Plasma vitamin E levels, however, tended to decrease as the type and quantity of PUFA in the diet increased. The present study illustrates that nutritionally beneficial PUFA in both fish and linseed oils are highly susceptible to biohydrogenation in the rumen. Although alpha-tocopheryl acetate resisted degradation in the rumen, plasma vitamin E status remained deficient to borderline, suggesting either that uptake may have been impaired or metabolism post-absorption increased.

  13. Bovine somatotropin and rumen-undegradable protein effects on skeletal growth in prepubertal dairy heifers.

    PubMed

    Moallem, U; Dahl, G E; Duffey, E K; Capuco, A V; Erdman, R A

    2004-11-01

    The objectives of this study were to determine the effects of dietary rumen-undegradable protein (RUP) and bovine somatotropin (bST) during the period from weaning until puberty on body weight (BW) and skeletal growth rates and age at puberty. Fifty-one Holstein heifers at 90 d of age were randomly assigned to 4 treatment groups consisting of 0.1 mg/kg BW per day of bST and 2% added dietary RUP (dry matter basis) applied in a 2 x 2 factorial design (n = 13 per group, except bST with no RUP group, n = 12). From 90 to 314 d, bST increased average daily gain (ADG) by 0.07 kg/d and BW by 16.2 kg, while added RUP increased ADG by 0.10 kg/d and BW by 21.4 kg. Both bST and added RUP effects on BW and ADG were additive. Skeletal growth rates, as measured by withers height (WH) and hip height (HH) were increased by both bST and added RUP. Somatotropin and RUP increased WH by 1.8 and 2.7 cm and hip height by 2.5 and 4.0 cm, respectively, at 314 d of age. Growth curves showed that added RUP effects on rates of BW, WH, and HH growth were greatest from 90 to 150 d age and diminished thereafter, suggesting that protein was limiting during this time period. Conversely, bST effects tended to be greater as the heifers approached puberty, but only in the presence of added RUP. Age at puberty was not affected by treatment, averaging 314 d of age across treatments. From 314 to 644 d of age, rates of BW, WH, and HH growth were similar among treatment groups. However, treatment differences present at 314 d of age persisted through 644 d of age, more than 10 mo after treatments ceased. These results suggest that protein during the early postweaning period and bST during the 200 to 300 d of age period just prior to puberty could be used to accelerate simultaneous increases in both BW and skeletal growth rates in dairy heifers without reducing age at puberty.

  14. Effects of corn silage and grass silage in ruminant rations on diurnal changes of microbial populations in the rumen of dairy cows.

    PubMed

    Lengowski, Melanie B; Witzig, Maren; Möhring, Jens; Seyfang, Gero M; Rodehutscord, Markus

    2016-12-01

    Here, we examined diurnal changes in the ruminal microbial community and fermentation characteristics of dairy cows fed total mixed rations containing either corn silage (CS) or grass silage (GS) as forage. The rations, which consisted of 52% concentrate and 48% GS or CS, were offered for ad libitum intake over 20 days to three ruminal-fistulated lactating Jersey cows during three consecutive feeding periods. Feed intake, ruminal pH, concentrations of short chain fatty acids and ammonia in rumen liquid, as well as abundance change in the microbial populations in liquid and solid fractions, were monitored in 4-h intervals on days 18 and 20. The abundance of total bacteria and Fibrobacter succinogenes increased in solids in cows fed CS instead of GS, and that of protozoa increased in both solid and liquid fractions. Feeding GS favored numbers of F. succinogenes and Selenomonas ruminantium in the liquid fraction as well as the numbers of Ruminobacter amylophilus, Prevotella bryantii and ruminococci in both fractions. Minor effects of silage were detected on populations of methanogens. Despite quantitative changes in the composition of the microbial community, fermentation characteristics were less affected by forage source. These results suggest a functional adaptability of the ruminal microbiota to total mixed rations containing either GS or CS as the source of forage. Diurnal changes in microbial populations were primarily affected by feed intake and differed between species and fractions, with fewer temporal fluctuations evident in the solid than in the liquid fraction. Interactions between forage source and sampling time were of minor importance to most of the microbial species examined. Thus, diurnal changes of microbial populations and fermentative activity were less affected by the two silages.

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

    PubMed

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

    2015-10-01

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

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

    PubMed Central

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

    2015-01-01

    This study aimed to investigate the effect of the ratio of non-fibrous carbohydrates to neutral detergent fibre (NFC/NDF) and undegraded dietary protein (UDP) on rumen fermentation and nitrogen metabolism in lambs. Four Dorper×thin-tailed Han crossbred lambs, averaging 62.3±1.9 kg of body weight and 10 mo of age, were randomly assigned to four dietary treatments of combinations of two levels of NFC/NDF (1.0 and 1.7) and two levels of UDP (35% and 50% of crude protein [CP]). Duodenal nutrient flows were measured with dual markers of Yb and Co, and microbial N (MN) synthesis was estimated using 15N. High UDP decreased organic matter (OM) intake (p = 0.002) and CP intake (p = 0.005). Ruminal pH (p<0.001), ammonia nitrogen (NH3-N; p = 0.008), and total volatile fatty acids (p<0.001) were affected by dietary NFC/NDF. The ruminal concentration of NH3-N was also affected by UDP (p<0.001). The duodenal flow of total MN (p = 0.007) was greater for lambs fed the high NFC/NDF diet. The amount of metabolisable N increased with increasing dietary NFC:NDF (p = 0.02) or UDP (p = 0.04). In conclusion, the diets with high NFC/NDF (1.7) and UDP (50% of CP) improved metabolisable N supply to lambs. PMID:26323398

  17. Tea saponins affect in vitro fermentation and methanogenesis in faunated and defaunated rumen fluid*

    PubMed Central

    Hu, Wei-lian; Wu, Yue-ming; Liu, Jian-xin; Guo, Yan-qiu; Ye, Jun-an

    2005-01-01

    The effect of tea saponins (TS) on rumen fermentation and methane emission was examined using an in vitro gas production technique named Reading Pressure Technique. Three levels of TS addition (0, 0.2, 0.4 mg/ml) were evaluated in the faunated and defaunated rumen fluid. Compared to the control, TS addition decreased the 24 h gas production in the faunated rumen fluid, but had a minor effect on gas yield in the defaunated rumen fluid. The TS significantly reduced methane production in vitro. In the faunated rumen fluid, 0.2 or 0.4 mg/ml TS decreased the 24 h methane emission by 12.7% or 14.0%, respectively. Rumen fluid pH value was affected neither by TS addition nor by defaunation. The TS addition had only minor effects on volatile fatty acids, but the yield and pattern of volatile fatty acids were greatly affected by defaunation. While the molar proportion of acetate was not affected by defaunation, the propionate was significantly increased and the butyrate significantly decreased. Ammonia-N concentration and microbial protein yield were influenced by TS inclusion and defaunation. Inclusion of 0.4 mg/ml TS increased the microbial protein mass by 18.4% and 13.8% and decreased the ammonia-N concentration by 8.3% and 19.6% in the faunated and defaunated rumen fluid, respectively. Protozoa counts were significantly reduced by TS inclusion. The current study demonstrated the beneficial effect of TS on methane production and rumen fermentation, and indicated that this may be due to the effect of the associated depression on protozoa counts. PMID:16052712

  18. Quantitative comparisons of select cultured and uncultured microbial populations in the rumen of cattle fed different diets.

    PubMed

    Kim, Minseok; Yu, Zhongtang

    2012-09-07

    The number and diversity of uncultured ruminal bacterial and archaeal species revealed by 16S rRNA gene (rrs) sequences greatly exceeds that of cultured bacteria and archaea. However, the significance of uncultured microbes remains undetermined. The objective of this study was to assess the numeric importance of select uncultured bacteria and cultured bacteria and the impact of diets and microenvironments within cow rumen in a comparative manner. Liquid and adherent fractions were obtained from the rumen of Jersey cattle fed hay alone and Holstein cattle fed hay plus grain. The populations of cultured and uncultured bacteria present in each fraction were quantified using specific real-time PCR assays. The population of total bacteria was similar between fractions or diets, while total archaea was numerically higher in the hay-fed Jersey cattle than in the hay-grain-fed Holstein cattle. The population of the genus Prevotella was about one log smaller than that of total bacteria. The populations of Fibrobacter succinogenes, Ruminococcus flavefaciens, the genus Butyrivibrio, and R. albus was at least one log smaller than that of genus Prevotella. Four of the six uncultured bacteria quantified were as abundant as F. succinogenes, R. flavefaciens and the genus Butyrivibrio. In addition, the populations of several uncultured bacteria were significantly higher in the adherent fractions than in the liquid fractions. These uncultured bacteria may be associated with fiber degradation. Some uncultured bacteria are as abundant as those of major cultured bacteria in the rumen. Uncultured bacteria may have important contribution to ruminal fermentation. Population dynamic studies of uncultured bacteria in a comparative manner can help reveal their ecological features and importance to rumen functions.

  19. Effects of one-seed juniper on intake, rumen fermentation, and plasma amino acids in sheep and goats fed supplemental protein

    USDA-ARS?s Scientific Manuscript database

    We tested the effect of feeding one-seed juniper on total intake, VFA profile, and plasma amino acids (AA) of 12 does and 12 ewes fed sudangrass and a basal diet with no protein supplement (Control; 5% CP) or rumen degradable (SBM; RDP 15% CP) or undegradable (FM; RUP 15% CP) protein supplement. Aft...

  20. Microbial ecology of the rumen evaluated by 454 GS FLX pyrosequencing is affected by starch and oil supplementation of diets.

    PubMed

    Zened, Asma; Combes, Sylvie; Cauquil, Laurent; Mariette, Jérôme; Klopp, Christophe; Bouchez, Olivier; Troegeler-Meynadier, Annabelle; Enjalbert, Francis

    2013-02-01

    To provide a comprehensive examination of the bacterial diversity in the rumen content of cows fed different diets, high-throughput 16S rRNA gene-based pyrosequencing was used. Four rumen fistulated nonlactating Holstein cows received 12 kg of dry matter per day of four diets based on maize silage during four periods: the low-starch diet (22% starch, 3% fat); the high-starch diet, supplemented with wheat plus barley (35% starch, 3% fat); the low-starch plus oil diet, supplemented with 5% of sunflower oil (20% starch, 7.6% fat) and the high-starch plus oil diet (33% starch, 7.3% fat). Samples were taken after 12 days of adaptation, 5 h postfeeding. Whatever the diet, bacterial community of sieved rumen contents was dominated by Firmicutes and Bacteroidetes. Lachnospiraceae, Ruminococcaceae, Prevotellaceae, and Rikenellaceae families were highly present and were clearly affected by cow diet. The highest abundance of Prevotellaceae and the lowest abundance of Ruminococcaceae and Rikenellaceae were found with the high-starch plus oil diet. Dietary starch increased the relative abundance of only three genera: Barnesiella, Oribacterium and Olsenella, but decreased the relative abundances of several genera, with very significant effects for Rikenellaceae_RC9 and Butyrivibrio-Pseudobutyrivibrio. Oil alone had a limited effect, but interestingly, starch plus oil addition differently affected the bacterial populations compared to starch addition without oil.

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

    PubMed

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

    2017-09-01

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

  2. Comparative Metabolite Fingerprinting of the Rumen System during Colonisation of Three Forage Grass (Lolium perenne L.) Varieties

    PubMed Central

    Kingston-Smith, Alison H.; Davies, Teri E.; Rees Stevens, Pauline; Mur, Luis A. J.

    2013-01-01

    The rumen microbiota enable ruminants to degrade complex ligno-cellulosic compounds to produce high quality protein for human consumption. However, enteric fermentation by domestic ruminants generates negative by-products: greenhouse gases (methane) and environmental nitrogen pollution. The current lack of cultured isolates representative of the totality of rumen microbial species creates an information gap about the in vivo function of the rumen microbiota and limits our ability to apply predictive biology for improvement of feed for ruminants. In this work we took a whole ecosystem approach to understanding how the metabolism of the microbial population responds to introduction of its substrate. Fourier Transform Infra Red (FTIR) spectroscopy-based metabolite fingerprinting was used to discriminate differences in the plant-microbial interactome of the rumen when using three forage grass varieties (Lolium perenne L. cv AberDart, AberMagic and Premium) as substrates for microbial colonisation and fermentation. Specific examination of spectral regions associated with fatty acids, amides, sugars and alkanes indicated that although the three forages were apparently similar by traditional nutritional analysis, patterns of metabolite flux within the plant-microbial interactome were distinct and plant genotype dependent. Thus, the utilisation pattern of forage nutrients by the rumen microbiota can be influenced by subtleties determined by forage genotypes. These data suggest that our interactomic approach represents an important means to improve forages and ultimately the livestock environment. PMID:24312434

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

    PubMed

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

    2009-12-01

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

  4. Effect of Carob Pod Extract on Cellulolysis, Proteolysis, Deamination, and Protein Biosynthesis in an Artificial Rumen1

    PubMed Central

    Tagari, H.; Henis, Y.; Tamir, Musha; Volcani, R.

    1965-01-01

    Carob pod extract and its tannin and sugar fractions were compared with gallotannic acid and sucrose for their effect on the cellulolytic, proteolytic, protein biosynthetic, and deaminative activities of rumen microorganisms. The inhibitory effects of carob pod extract upon the cellulolysis and deamination were correlated mainly with its sugar, rather than its tannin components. On the other hand, proteolytic activity and protein biosynthesis were more significantly affected by the tannin fraction. In contrast to the tannin fraction of carob pod extract, gallotannic acid inhibited cellulolytic activity. The harmful effect of a low concentration of tannins on protein biosynthesis could be prevented by the addition of carbohydrates to the reaction mixture. At high tannin concentration (40 μg/ml), however, the addition of carbohydrates did not prevent the inhibition. PMID:14325284

  5. Metagenomic Insights into the RDX-Degrading Potential of the Ovine Rumen Microbiome

    PubMed Central

    Li, Robert W.; Giarrizzo, Juan Gabriel; Wu, Sitao; Li, Weizhong; Duringer, Jennifer M.; Craig, A. Morrie

    2014-01-01

    The manufacturing processes of royal demolition explosive (RDX), or hexahydro-1,3,5-trinitro-1,3,5-triazine, have resulted in serious water contamination. As a potential carcinogen, RDX can cause a broad range of harmful effects to humans and animals. The ovine rumen is capable of rapid degradation of nitroaromatic compounds, including RDX. While ruminal RDX-degrading bacteria have been identified, the genes and pathways responsible for RDX degradation in the rumen have yet to be characterized. In this study, we characterized the metabolic potential of the ovine rumen using metagenomic approaches. Sequences homologous to at least five RDX-degrading genes cloned from environmental samples (diaA, xenA, xenB, xplA, and xplB) were present in the ovine rumen microbiome. Among them, diaA was the most abundant, likely reflective of the predominance of the genus Clostridium in the ovine rumen. At least ten genera known to harbor RDX-degrading microorganisms were detectable. Metagenomic sequences were also annotated using public databases, such as Pfam, COG, and KEGG. Five of the six Pfam protein families known to be responsible for RDX degradation in environmental samples were identified in the ovine rumen. However, increased substrate availability did not appear to enhance the proliferation of RDX-degrading bacteria and alter the microbial composition of the ovine rumen. This implies that the RDX-degrading capacity of the ovine rumen microbiome is likely regulated at the transcription level. Our results provide metagenomic insights into the RDX-degrading potential of the ovine rumen, and they will facilitate the development of novel and economic bioremediation strategies. PMID:25383623

  6. Metagenomic insights into the RDX-degrading potential of the ovine rumen microbiome.

    PubMed

    Li, Robert W; Giarrizzo, Juan Gabriel; Wu, Sitao; Li, Weizhong; Duringer, Jennifer M; Craig, A Morrie

    2014-01-01

    The manufacturing processes of royal demolition explosive (RDX), or hexahydro-1,3,5-trinitro-1,3,5-triazine, have resulted in serious water contamination. As a potential carcinogen, RDX can cause a broad range of harmful effects to humans and animals. The ovine rumen is capable of rapid degradation of nitroaromatic compounds, including RDX. While ruminal RDX-degrading bacteria have been identified, the genes and pathways responsible for RDX degradation in the rumen have yet to be characterized. In this study, we characterized the metabolic potential of the ovine rumen using metagenomic approaches. Sequences homologous to at least five RDX-degrading genes cloned from environmental samples (diaA, xenA, xenB, xplA, and xplB) were present in the ovine rumen microbiome. Among them, diaA was the most abundant, likely reflective of the predominance of the genus Clostridium in the ovine rumen. At least ten genera known to harbor RDX-degrading microorganisms were detectable. Metagenomic sequences were also annotated using public databases, such as Pfam, COG, and KEGG. Five of the six Pfam protein families known to be responsible for RDX degradation in environmental samples were identified in the ovine rumen. However, increased substrate availability did not appear to enhance the proliferation of RDX-degrading bacteria and alter the microbial composition of the ovine rumen. This implies that the RDX-degrading capacity of the ovine rumen microbiome is likely regulated at the transcription level. Our results provide metagenomic insights into the RDX-degrading potential of the ovine rumen, and they will facilitate the development of novel and economic bioremediation strategies.

  7. Comparison of fermentation of diets of variable composition and microbial populations in the rumen of sheep and Rusitec fermenters. I. Digestibility, fermentation parameters, and microbial growth.

    PubMed

    Martínez, M E; Ranilla, M J; Tejido, M L; Ramos, S; Carro, M D

    2010-08-01

    Four ruminally and duodenally cannulated sheep and 8 Rusitec fermenters were used to determine the effects of forage to concentrate (F:C) ratio and type of forage in the diet on ruminal fermentation and microbial protein synthesis. The purpose of the study was to assess how closely fermenters can mimic the dietary differences found in vivo. The 4 experimental diets contained F:C ratios of 70:30 or 30:70 with either alfalfa hay or grass hay as the forage. Microbial growth was determined in both systems using (15)N as a microbial marker. Rusitec fermenters detected differences between diets similar to those observed in sheep by changing F:C ratio on pH; neutral detergent fiber digestibility; total volatile fatty acid concentrations; molar proportions of acetate, propionate, butyrate, isovalerate, and caproate; and amylase activity. In contrast, Rusitec fermenters did not reproduce the dietary differences found in sheep for NH(3)-N and lactate concentrations, dry matter (DM) digestibility, proportions of isobutyrate and valerate, carboxymethylcellulase and xylanase activities, and microbial growth and its efficiency. Regarding the effect of the type of forage in the diet, Rusitec fermenters detected differences between diets similar to those found in sheep for most determined parameters, with the exception of pH, DM digestibility, butyrate proportion, and carboxymethylcellulase activity. Minimum pH and maximal volatile fatty acid concentrations were reached at 2h and at 6 to 8h postfeeding in sheep and fermenters, respectively, indicating that feed fermentation was slower in fermenters compared with that in sheep. There were differences between systems in the magnitude of most determined parameters. In general, fermenters showed lower lactate concentrations, neutral detergent fiber digestibility, acetate:propionate ratios, and enzymatic activities. On the contrary, fermenters showed greater NH(3)-N concentrations, DM digestibility, and proportions of propionate

  8. Measurement of the Intestinal Digestibility of Rumen Undegraded Protein Using Different Methods and Correlation Analysis.

    PubMed

    Wang, Y; Zhang, Y G; Liu, Xiaolan; Kopparapu, N K; Xin, Hangshu; Liu, J; Guo, Jianhua

    2015-10-01

    Four methods were adopted, including the mobile nylon bag (MNB) method, modified three-step in vitro (MTS) method, original three-step in vitro (OTS) method, and acid detergent insoluble nitrogen (ADIN) estimating method, to evaluate the intestinal digestibility of rumen undegradable protein (DRUP) of 10 types of concentrates and 7 types of roughages. After correlation analysis to determine the DRUP values using the MNB, MTS, OTS, and ADIN methods, the study aimed to find out appropriate methods to replace the MNB method due to its disadvantages such as high price, long time period, and use of a duodenal T-fistula. Three dairy cows with a permanent ruminal fistula and duodenal T-fistula were used in a single-factor experimental design. The results showed that the determined DRUP values using the MNB method for soybean meal, cottonseed meal, rapeseed meal, sunflower meal, corn germ meal, corn, rice bran, barley, wheat bran, corn fiber feed, Alfalfa (Zhao dong), Alfalfa (Long mu 801), Alfalfa (Long mu 803), grass (North), Grass (Inner Mongolia), corn silage and corn straw were 98.13%, 87.37%, 88.47%, 82.60%, 75.40%, 93.23%, 69.27%, 91.27%, 72.37%, 79.03%, 66.72%, 68.64%, 73.57%, 50.47%, 51.52%, 54.05%, and 43.84%, respectively. The coefficient of determination (R (2) = 0.964) of the results between the MTS method and the MNB method was higher than that (R (2) = 0.942) between the OTS method and the MNB method. The coefficient of determination of the DRUP values of the concentrates among the in vitro method (including the MTS and OTS methods) and the MNB method was higher than that of the roughage. There was a weak correlation between the determined DRUP values in concentrates obtained from the ADIN method and those from the MNB method, and there was a significant correlation (p<0.01) between the determined DRUP values of the roughage obtained from the MNB method and those obtained from ADIN method. The DRUP values were significantly correlated with the nutritional

  9. Effects of dietary supplementing tannic acid in the ration of beef cattle on rumen fermentation, methane emission, microbial flora and nutrient digestibility.

    PubMed

    Yang, K; Wei, C; Zhao, G Y; Xu, Z W; Lin, S X

    2017-04-01

    Four adult Simmental male cattle (376 ± 9.0 kg initial BW), fitted with permanent rumen cannulas, were used in a 4 × 4 Latin square design to investigate the effects of dietary supplementing tannic acid (TA) on rumen fermentation, methane (CH4 ) production, rumen microbes, nutrient digestibility and plasma biochemical parameters. Four levels of TA, that is 0, 6.5, 13.0 or 26.0 g/kg dry matter (DM), were added to the basal ration (composed of corn silage and concentrate mixture) as experimental treatments respectively. Each experimental period consisted of a 12-day adaptation phase followed by a 3-day sampling phase. The results showed that supplementing TA at 26.0 g/kg DM decreased the relative abundance of protozoa, methanogens and Ruminococcus albus to the total ruminal bacterial 16S rDNA in beef cattle (p < 0.05). The results also showed that supplementing TA at 6.5, 13.0 or 26.0 g/kg DM decreased (p < 0.01) the CH4 production (l/kg DM intake) by 11.1%, 14.7% and 33.6% respectively. Supplementing TA at 13.0 or 26.0 g/kg DM decreased the ratio of acetate to propionate and ammonia nitrogen (NH3 -N) (p < 0.05) and tended to decrease the total volatile fatty acid (VFA) concentration of rumen fluid (p = 0.07). Supplementing TA at 26.0 g/kg DM decreased DM and organic matter (OM) digestibility (p < 0.05), supplementing TA at 6.5, 13.0 or 26.0 g/kg DM decreased (p < 0.01) crude protein (CP) digestibility by 5.0%, 8.6% and 15.7%, respectively, and supplementing TA at 6.5, 13.0 or 26.0 g/kg DM increased (p < 0.05) the plasma total antioxidant capability. It was concluded that supplementing TA in the ration of beef cattle decreased the CH4 production and digestibility of CP of beef cattle. Supplementing TA could be an effective option to mitigate CH4 emission form cattle, further research is necessary to study the effects of TA on the performance of cattle.

  10. The effects of ruminally degraded protein on rumen fermentation and ammonia losses from manure in dairy cows.

    PubMed

    Agle, M; Hristov, A N; Zaman, S; Schneider, C; Ndegwa, P; Vaddella, V K

    2010-04-01

    This experiment investigated the effect of dietary crude protein (CP) and ruminally degraded protein (RDP) levels on rumen fermentation, digestibility, ammonia emission from manure, and performance of lactating dairy cows. The experiment was a replicated 3 x 3 Latin square design with 6 cows. Three diets varying in CP concentration were tested (CP, % of dry matter): 15.4 (high CP, control), 13.4 (medium CP), and 12.9% (low CP). These diets provided metabolizable protein balances of 323, -44, and 40 g/d and RDP balances of 162, -326, and -636 g/d (high, medium, and low, respectively). Both the medium and low CP diets decreased ruminal pH compared with high CP, most likely because of the higher nonfiber carbohydrate concentration in the former diets. Ruminal ammonia pool size (rumen ammonia N was labeled with (15)N) and the concentration of total free amino acids were greater for the high CP diet than for the RDP-deficient diets. Apparent total-tract nutrient digestibilities were not affected by treatment. Both the medium and low CP diets resulted in lower absolute and relative excretion of urinary N compared with the high CP diet, as a proportion of N intake. Excretion of fecal N and milk yield and composition were not affected by diet. Milk N efficiency (milk N / N intake) and the cumulative secretion of ammonia-(15)N in milk protein were greater for the RDP-deficient diets, and milk urea N concentration was greater for the high CP diet. Both medium and low CP diets decreased the irreversible loss of ruminal ammonia N compared with the high CP diet. The rate and cumulative ammonia emissions from manure were lower for the medium and low CP diets compared with the high CP diet. Overall, this study demonstrated that dairy diets with reduced CP and RDP concentrations will produce manure with lower ammonia-emitting potential without affecting cow performance, if metabolizable protein requirements are met.

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

    PubMed

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

    2017-03-08

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

  12. Manipulation of rumen ecology by dietary lemongrass (Cymbopogon citratus Stapf.) powder supplementation.

    PubMed

    Wanapat, M; Cherdthong, A; Pakdee, P; Wanapat, S

    2008-12-01

    This experiment was conducted to investigate the effect of lemongrass [Cymbopogon citratus (DC.) Stapf.] powder (LGP) on rumen ecology, rumen microorganisms, and digestibility of nutrients. Four ruminally fistulated crossbred (Brahman native) beef cattle were randomly assigned according to a 4 x 4 Latin square design. The dietary treatments were LGP supplementation at 0, 100, 200, and 300 g/d with urea-treated rice straw (5%) fed to allow ad libitum intake. Digestibilities of DM, ether extract, and NDF were significantly different among treatments and were greatest at 100 g/d of supplementation. However, digestibility of CP was decreased with LGP supplementation (P < 0.05), whereas ruminal NH(3)-N and plasma urea N were decreased with incremental additions of LGP (P < 0.05). Ruminal VFA concentrations were similar among supplementation concentrations (P > 0.05). Total viable bacteria, amylolytic bacteria, and cellulolytic bacteria were significantly different among treatments and were greatest at 100 g/d of supplementation (4.7 x 10(9), 1.7 x 10(7), and 2.0 x 10(9) cfu/mL, respectively). Protozoal populations were significantly decreased by LGP supplementation. In addition, efficiency of rumen microbial N synthesis based on OM truly digested in the rumen was enriched by LGP supplementation, especially at 100 g/d (34.2 g of N/kg of OM truly digested in the rumen). Based on this study, it could be concluded that supplementation of LGP at 100 g/d improved digestibilities of nutrients, rumen microbial population, and microbial protein synthesis efficiency, thus improving rumen ecology in beef cattle.

  13. Rumen-degradable protein in roll conditioned or macerated legume hays and silages estimated by in situ kinetics vs. alternative methods

    USDA-ARS?s Scientific Manuscript database

    Alternatives to the in situ method for estimating rumen-degradable protein (RDP) in diverse forage legumes should be validated. In this study, RDP in roll conditioned or macerated silages and hays of Medicago, Lotus, and Trifolium species with differing polyphenol compositions were estimated from in...

  14. Supplementing rumen-protected methionine and lysine in low-protein diets based on corn distillers grains fed to lactating dairy cows

    USDA-ARS?s Scientific Manuscript database

    Feeding rumen-protected methionine (RPM) and lysine (RPL) may allow feeding lower crude protein (CP) diets to dairy cows, thereby increasing nitrogen efficiency and reducing environmental impact. Moreover, RPL supplementation may improve the value of corn distillers dried grains plus solubles (DDGS)...

  15. Brazilian propolis extract used as an additive to decrease methane emissions from the rumen microbial population in vitro.

    PubMed

    Santos, Nadine Woruby; Zeoula, Lucia Maria; Yoshimura, Emerson Henri; Machado, Erica; Macheboeuf, Didier; Cornu, Agnès

    2016-06-01

    Propolis is a product that is rich in phenolic compounds and can be utilized in animal nutrition as a dietary additive. In this study, the effects of a Brazilian green propolis extract on rumen fermentation and gas production were determined. The fate of propolis phenolic compounds in the rumen medium was also investigated. Fermentation was done in 24-h batches over three periods. Inoculates were obtained from cows fed on grassland hay and concentrate. Propolis extract in a hydroalcoholic solution was applied at increasing doses to the substrate (1 to 56 g/kg). The fermentation substrate consisted on a mixture of alfalfa hay, soybean meal, and wheat grain mixture in dry matter. After 24 h of fermentation, seven new compounds were observed in the medium in amounts that correlated to the propolis dose. The dose of propolis extract linearly decreased the pH of the medium and linearly increased propionate production, which reduced the acetate-to-propionate ratio and influenced the total production of short-chain fatty acids. Propolis also linearly reduced methane production and increased the carbon dioxide-to-methane ratio. Ammonia nitrogen levels and in vitro digestibility of organic matter were similar among the treatments. The combination of increased propionate production and decreased methane production suggests better energy utilization from the feed.

  16. Effect of monensin on in vitro fermentation of silages and microbial protein synthesis.

    PubMed

    Wischer, Gerald; Boguhn, Jeannette; Steingaß, Herbert; Schollenberger, Margit; Hartung, Karin; Rodehutscord, Markus

    2013-06-01

    The objective of the study was to investigate the effects of monensin on silage fermentation and microbial net protein synthesis. In Experiment 1, monensin (0.5, 1, 2, 4, 6, or 10 µg) was added to syringes that contained 120 mg of grass silage (GS), grass silage and concentrate (GS + C), or maize silage (MS), resulting in concentrations of 4.2, 8.3, 16.7, 33.3, 50.0 and 83.3 mg monensin/kg feed. Samples were incubated for 24 h to determine the monensin concentration that resulted in the maximum reduction in methane production without effects on the total gas production. In Experiment 2, GS and GS + C were incubated in a rumen simulation technique (Rusitec) to assess the monensin effects (133 and 266 mg/kg feed) on the production of total gas, methane and volatile fatty acids (VFA), degradation of nutrients and microbial net protein synthesis. In Experiment 1, methane production was reduced without significant effects on the total gas production; the reductions were 17% (GS), 10% (GS + C) and 13% (MS) with 16.7 (GS), 50.0 (GS + C) and 33.3 (MS) mg monensin/kg feed. Monensin reduced the total gas and methane production in GS and GS + C in Experiment 2. Propionate production was enhanced by monensin, accompanied by a decrease in acetate production. Along with a reduction in crude protein (CP) degradation, monensin reduced the ammonia nitrogen concentration in the effluent of both treatments. While the protein produced by liquid-associated microbes increased with monensin, protein production by solid-associated microbes was reduced. Total microbial net protein synthesis increased in the presence of monensin. Monensin influenced the production of total gas, methane and VFA from the silages without an effect on the degradation of organic matter (OM). Different microbial fractions were affected differently by monensin supplementation. If monensin is used as a tool to reduce methane emission, the supplementation level must be carefully chosen to avoid negative effects on

  17. Determination of succession of rumen bacterial species in nursing beef calves

    USDA-ARS?s Scientific Manuscript database

    Ruminants are typically born with a non-functional rumen essentially devoid of microorganisms. The succession of the microbial population in the rumen from birth to animal maturity is of interest due to the key role that the rumen microbial population plays in the overall health and productivity of ...

  18. Perspectives on ruminant nutrition and metabolism I. Metabolism in the rumen.

    PubMed

    Annison, E F; Bryden, W L

    1998-12-01

    Advances in knowledge of ruminant nutrition and metabolism during the second half of the twentieth century have been reviewed. Part I is concerned with metabolism in the rumen: Part II discusses utilization of nutrients absorbed from the rumen and lower tract to support growth and reproduction. The time frame was prompted by the crucial advances in ruminant physiology which arose from the work of Sir Jospeh Barcroft and his colleagues at Cambridge in the 1940s and 50s, and by the brilliant studies of Robert Hungate on rumen microbiology at much the same time. In reviewing the growth of knowledge of the role of bacteria, protozoa, fungi and bacteriophages in the rumen, outstanding developments have included the identification and characterization of fungi and the recognition that the utilization of polysaccharides in the rumen is accomplished by the sequential activities of consortia of rumen microorganisms. The role of protozoa is discussed in relation to the long standing debate on whether or not the removal of protozoa (defaunation) improves the efficiency of ruminant production. In relation to nitrogen (N) metabolism, the predation of bacteria by protozoa increases protein turnover in the rumen and reduces the efficiency of microbial protein production. This may account for the beneficial effects of defaunation where dietary N intakes are low and possibly rate limiting for growth and production. Current approaches to the measurement of rates of production of short chain fatty acids (SCFA) in the rumen based on the mathematical modelling of isotope dilution data are outlined. The absorption of SCFA from the rumen and hindgut is primarily a passive permeation process. The role of microorganisms in N metabolism in the rumen has been discussed in relation to ammonia and urea interrelationships and to current inadequacies in the measurement of both protein degradation in the rumen and microbial protein synthesis. The growth of knowledge of digestion and absorption of

  19. Effects of dietary protein on milk, rumen, and blood parameters in dairy cattle fed low fiber diets.

    PubMed

    Jaquette, R D; Rakes, A H; Croom, W J

    1986-04-01

    Eighteen multiparous and 9 primiparous Holstein cows were used to determine the effects of a 13 and 23% crude protein concentrate on milk fat depression during early lactation. Beginning on d 22 postpartum, cows were fed a high fiber diet (27% acid detergent fiber) for 3 wk and then switched to a low fiber diet (9 to 10% acid detergent fiber) for 6 wk. Crude protein percentages calculated from dry matter consumption were 13.5 and 17.9% during the high fiber period and 12.7 and 22.3% during the low fiber period. Daily milk and fat yields for both primiparous and multiparous cows were greater for the high protein treatment. The magnitude of decline in milk fat percentage (from high to low fiber) was greater for the low protein treatment, as determined by nonlinear regression. The high protein treatment was more effective in reducing the severity of milk fat depression in primiparous cows than in multiparous cows. Dietary crude protein had no effect on milk protein or solids-not-fat percentages, rumen volatile fatty acid molar proportions, or serum acetate concentration. The mechanism by which the high protein ration minimized the fat depression response to low fiber rations by primiparous cows is unknown.

  20. Effect of donor animals and their diet on in vitro nutrient degradation and microbial protein synthesis using grass and corn silages.

    PubMed

    Boguhn, J; Zuber, T; Rodehutscord, M

    2013-06-01

    Two nonlactating cows and two wether sheep, all fitted with a permanent cannula into the rumen, were fed either hay plus concentrate, grass silage or corn silage to study the effect of the donor animal and its diet on in vitro fermentation and microbial protein synthesis. Rumen inoculum was obtained before the morning feeding. Grass silage or corn silage was incubated in a semi-continuous rumen simulation system for 14 days. Four replicated vessels were used per treatment. Degradation of crude nutrients and detergent fibre fractions as well as microbial protein synthesis and the production of volatile fatty acids were studied. Additionally, total gas and methane production was measured with a standard in vitro gas test. Gas production and methane concentration was higher when the inoculum used was from sheep than that from cows. The donor animal also affected the degradation of organic matter and ether extract as well as the amount of propionate and butyrate, and the acetate-to-propionate ratio. The effect of the diet fed to the donor animal on fermentation was much greater than the effect of the donor animal itself. Feeding hay plus concentrate resulted in higher gas production and degradation of acid detergent fibre, but in lower degradation of ether extract and reduced microbial protein synthesis. Additionally, the pattern of volatile fatty acids changed significantly when the diet of the donor animals was hay plus concentrate or one of the silages. These results show that in vitro fermentation and microbial protein synthesis is different when based on inoculum from either cattle or sheep. The diet fed to the donor animal is more important than the animal species and is probably mediated by an adjusted microbial activity. With regard to standardized feed evaluations, these results further support the need to harmonize in vitro approaches used in different laboratories.

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

    PubMed

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

    2011-01-01

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

  2. Recovery and upgrading bovine rumen protein by extrusion: effect of lipid content on protein disulphide cross-linking, solubility and molecular weight.

    PubMed

    Vaz, L C M A; Arêas, J A G

    2010-01-01

    Bovine rumen protein with two levels of residual lipids (1.9% or 3.8%) was subjected to thermoplastic extrusion under different temperatures and moisture contents. Protein solubility in different buffers, disulphide cross-linking and molecular weight distribution were determined on the extrudates. After extrusion, samples with 1.9% residual lipids content had a higher concentration of protein insoluble by undetermined forces, irrespective of feed moisture and processing temperature used. Lipid content of 3.8% in the feed material resulted in more protein participating in the extrudate network through non-covalent interactions (hydrophobic and electrostatic) and disulphide bonds. A small dependency of the extrusion process on moisture and temperature and a marked dependency on lipid content, especially phospholipid, was observed, Electrophoresis under non-reducing conditions showed that protein extrusion with low feed moisture promoted high molecular breakdown inside the barrel, probably due to intense shear force, and further protein aggregation at the die end.

  3. Pelleted beet pulp substituted for high-moisture corn: 3. Effects on ruminal fermentation, pH, and microbial protein efficiency in lactating dairy cows.

    PubMed

    Voelker, J A; Allen, M S

    2003-11-01

    The effects of increasing concentrations of dried, pelleted beet pulp substituted for high-moisture corn on ruminal fermentation, pH, and microbial efficiency were evaluated using eight ruminally and duodenally cannulated multiparous Holstein cows in a duplicated 4 x 4 Latin square design with 21-d periods. Cows were 79 +/- 17 (mean +/- SD) DIM at the beginning of the experiment. Experimental diets with 40% forage (corn silage and alfalfa silage) and 60% concentrate contained 0, 6.1, 12.1, or 24.3% beet pulp substituted for high-moisture corn on a DM basis. Diet concentrations of NDF and starch were 24.3 and 34.6% (0% beet pulp), 26.2 and 30.5% (6% beet pulp), 28.0, and 26.5% (12% beet pulp), and 31.6 and 18.4% (24% beet pulp), respectively. Substituting beet pulp for corn did not affect daily mean or minimum ruminal pH but tended to reduce pH range. Ruminal acetate:propionate responded in a positive exponential relationship to added beet pulp. Rate of valerate absorption from the rumen was not affected by treatment. Substituting beet pulp for corn up to 24% of diet DM did not affect efficiency of ruminal microbial protein production, expressed as microbial N flow to the duodenum as a percentage of OM truly digested in the rumen. Microbial efficiency was not correlated to mean pH or daily minimum pH. While microbial efficiency was not directly related to concentration of beet pulp fed, it was positively correlated with passage rate of particulate matter, as represented by starch and indigestible NDF, probably due to reduced turnover of microbial protein in the rumen.

  4. Metagenomics detects functional shifts in the bovine rumen microbiota in response to propionate intake

    USDA-ARS?s Scientific Manuscript database

    Optimizing rumen microbial ecosystem output is essential towards improved ruminant agriculture. Ruminal infusion or intake of propionate, one of the predominant volatile fatty acids, has important implications for host physiology. However, how the rumen microbiota responds to propionate administrat...

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

    PubMed

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

    2017-02-01

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

  6. Rumen Microbiology

    USDA-ARS?s Scientific Manuscript database

    Ruminant animals such as cattle, sheep, and goats are able to digest low-quality fibrous feedstuffs because they maintain a mutually beneficial relationship with microorganisms resident in their forestomach, the rumen. Ruminal microorganisms are bacteria, protozoa, fungi, and viruses that live in a...

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

    PubMed

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

    2007-06-01

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

  8. Exploring the sheep rumen microbiome for carbohydrate-active enzymes.

    PubMed

    Lopes, Lucas Dantas; de Souza Lima, André Oliveira; Taketani, Rodrigo Gouvêa; Darias, Phillip; da Silva, Lília Raquel Fé; Romagnoli, Emiliana Manesco; Louvandini, Helder; Abdalla, Adibe Luiz; Mendes, Rodrigo

    2015-07-01

    The rumen is a complex ecosystem enriched for microorganisms able to degrade biomass during the animal's digestion process. The recovery of new enzymes from naturally evolved biomass-degrading microbial communities is a promising strategy to overcome the inefficient enzymatic plant destruction in industrial production of biofuels. In this context, this study aimed to describe the bacterial composition and functions in the sheep rumen microbiome, focusing on carbohydrate-active enzymes (CAE). Here, we used phylogenetic profiling analysis (inventory of 16S rRNA genes) combined with metagenomics to access the rumen microbiome of four sheep and explore its potential to identify fibrolytic enzymes. The bacterial community was dominated by Bacteroidetes and Firmicutes, followed by Proteobacteria. As observed for other ruminants, Prevotella was the dominant genus in the microbiome, comprising more than 30 % of the total bacterial community. Multivariate analysis of the phylogenetic profiling data and chemical parameters showed a positive correlation between the abundance of Prevotellaceae (Bacteroidetes phylum) and organic matter degradability. A negative correlation was observed between Succinivibrionaceae (Proteobacteria phylum) and methane production. An average of 2 % of the shotgun metagenomic reads was assigned to putative CAE when considering nine protein databases. In addition, assembled contigs allowed recognition of 67 putative partial CAE (NCBI-Refseq) representing 12 glycosyl hydrolase families (Pfam database). Overall, we identified a total of 28 lignocellulases, 22 amylases and 9 other putative CAE, showing the sheep rumen microbiome as a promising source of new fibrolytic enzymes.

  9. Using a complex non-TDN based model (the DVE/OEB system) to predict microbial protein synthesis, endogenous protein, degradation balance, and total truly absorbed protein supply of different varieties of cereal oats for ruminants.

    PubMed

    Yu, Peiqiang; Niu, Zhiyuan

    2009-06-01

    Recently a new super-genotype of oat has been developed in the Crop Development Center called CDC SO-I ('SuperOat': low lignin and high fat). In a previous study, we evaluated total metabolizable protein using a TDN-based model-NRC-2001 which is popular in North America. However, the TDN-based NRC model is not accepted universally. The objectives of this study were to use a complex non-TDN based model, the DVE/OEB system, to evaluate potential nutrient supply to ruminants from the SuperOat in comparison with two normal varieties of oats (CDC Dancer and Derby) in western Canada. The quantitative predictions were made in terms of: (i) truly absorbed rumen synthesized microbial proteins in the small intestine; (ii) truly absorbed rumen undegraded feed protein in the small intestine; (iii) endogenous protein in the digestive tract; (iv) total truly absorbed protein in the small intestine; and (v) protein degraded balance. Results showed that using the DVE/OEB system to predict the potential nutrient supply, it was found that the SuperOat had similar truly absorbed rumen synthesized microbial protein levels (61, 63, 59 g/kg DM, P > 0.05, for SuperOat, CDC Dancer and Derby, respectively), higher truly absorbed rumen undegraded feed protein than CDC Dancer (22 vs. 17 g/kg DM P < 0.05, for SuperOat, CDC Dancer, respectively), but similar to Derby (22 vs. 21 g/kg DM; P > 0.05), and similar endogenous protein (16, 16, 18 g/kg DM; P > 0.05). Total truly absorbed protein in the small intestine is only numerically higher in the SuperOat (67 vs. 65, 62 g/kg DM, P > 0.05, for CDC Dancer and Derby, respectively). However, the protein degraded balance was significantly different (P < 0.05) with a positive value for the SuperOat (7.0 g/kg DM) and negative values for two normal varieties (-1.5, -6.8 g/kg DM for CDC Dancer and Derby, respectively). In conclusion, the model predicted significantly different protein degradation balance. The SuperOat had positive degradation balance but

  10. Supplementation of methionine and selection of highly digestible rumen undegradable protein to improve nitrogen efficiency for milk production.

    PubMed

    Noftsger, S; St-Pierre, N R

    2003-03-01

    Metabolizable protein (MP) supply and amino acid balance were manipulated through selection of highly digestible rumen-undegradable protein (RUP) sources and methionine (Met) supplementation. Effects on production efficiency and N utilization of lactating dairy cows were determined. Thirty-two multiparous (647 kg) and 28 primiparous (550 kg) Holstein cows were assigned during the fourth week of lactation to one of four dietary treatments. Treatments were 1) 18.3% crude protein (CP) with low estimated intestinal digestibility of RUP (HiCP-LoDRUP), 2) 18.3% CP with high digestibility RUP (HiCP-HiDRUP), 3) 16.9% CP with high digestibility RUP (LoCP-HiDRUP), and 4) 17.0% CP with high digestibility RUP and supplemental Met (LoCP-HiDRUP + Met). Diets were balanced to have equal concentrations of net energy for lactation (NE(L)), acid detergent fiber (ADF), neutral detergent fiber (NDF), and ash. Milk yields (40.8, 46.2, 42.9, 46.6 kg/d), protein percentages (2.95, 2.98, 2.99, 3.09%), and fat percentages (3.42, 3.64, 3.66, 3.73%) are reported here for HiCP-LoDRUP, HiCP-HiDRUP, LoCP-HiDRUP, and LoCP-HiDRUP + Met, respectively. Milk urea N and BUN decreased when feeding a lower CP diet. Efficiency of use of N for milk protein production was higher when feeding higher digestibility RUP, especially with the LoCP-HiDRUP + Met diet. A digestibility study followed the production trial, with six cows per treatment group continuing on the same treatment for an additional week. The experimental periods were 5 d long, with 1 d of adjustment and 4 d of total collection of urine and feces. Dry matter intake, milk production, milk protein production, and N digestibility were not significantly different among treatments during the collection trial, whereas N intake and N absorbed increased with the higher CP diets. The quantity of N in feces did not change with diet, but quantity of N in urine decreased in the low CP diets. Milk N as a percentage of intake N and milk N as a percentage

  11. Protease activities of rumen protozoa.

    PubMed Central

    Forsberg, C W; Lovelock, L K; Krumholz, L; Buchanan-Smith, J G

    1984-01-01

    Intact, metabolically active rumen protozoa prepared by gravity sedimentation and washing in a mineral solution at 10 to 15 degrees C had comparatively low proteolytic activity on azocasein and low endogenous proteolytic activity. Protozoa washed in 0.1 M potassium phosphate buffer (pH 6.8) at 4 degrees C and stored on ice autolysed when they were warmed to 39 degrees C. They also exhibited low proteolytic activity on azocasein, but they had a high endogenous proteolytic activity with a pH optimum of 5.8. The endogenous proteolytic activity was inhibited by cysteine proteinase inhibitors, for example, iodoacetate (63.1%) and the aspartic proteinase inhibitor, pepstatin (43.9%). Inhibitors specific for serine proteinases and metalloproteinases were without effect. The serine and cysteine proteinase inhibitors of microbial origin, including antipain, chymostatin, and leupeptin, caused up to 67% inhibition of endogenous proteolysis. Hydrolysis of casein by protozoa autolysates was also inhibited by cysteine proteinase inhibitors. Some of the inhibitors decreased endogenous deamination, in particular, phosphoramidon, which had little inhibitory effect on proteolysis. Protozoal and bacterial preparations exhibited low hydrolytic activities on synthetic proteinase and carboxypeptidase substrates, although the protozoa had 10 to 78 times greater hydrolytic activity (per milligram of protein) than bacteria on the synthetic aminopeptidase substrates L-leucine-p-nitroanilide, L-leucine-beta-naphthylamide, and L-leucinamide. The aminopeptidase activity was partially inhibited by bestatin. It was concluded that cysteine proteinases and, to a lesser extent, aspartic proteinases are primarily responsible for proteolysis in autolysates of rumen protozoa. The protozoal autolysates had high aminopeptidase activity; low deaminase activity was observed on endogenous amino acids. PMID:6364968

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

    PubMed

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

    2009-09-01

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

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

    PubMed

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

    2016-02-01

    The goals of ruminant protein nutrition are to provide adequate amounts of RDP for optimal ruminal efficiency and to obtain the desired animal productivity with a minimum amount of dietary CP. The aim of the present study was to examine effects of decreasing dietary protein by decreasing RDP with the optimum concentration of RUP on production performance, nutrient digestibility, N retention, rumen fermentation parameters, and blood metabolites in high-producing Holstein cows in early lactation. Nine multiparous lactating cows (second parities, averaging 50 ± 12 d in milk and milk yield of 48 ± 5 kg/d) were used in a triplicate 3 × 3 Latin square design with 3 rations: 1) a total mixed ration (TMR) containing 16.4% CP (10.9% RDP based on DM), 2) a TMR containing 15.6% CP (10% RDP), and 3) a TMR containing 14.8% CP (9.3% RDP). The level of RUP was constant at 5.5% DM across the treatments. All diets were calculated to supply a postruminal lysine to methionine ratio of about 3:1. Dry matter intake, milk yield and composition, 4% fat-corrected milk, and energy-corrected milk were not significantly affected by decreasing dietary CP and RDP levels. Cows fed 16.4% CP diets had greater ( < 0.01) CP and RDP intakes, which resulted in a trend toward greater concentrations of plasma urea N compared with other treatments. Daily N intake linearly decreased ( < 0.01) with decreasing dietary CP and RDP levels, whereas the intake of RUP and fecal N excretion (g/d) did not change. Apparent digestibility of nutrients, ruminal pH, and NH-N concentration were not affected with decreasing dietary CP and RDP levels. Apparent N efficiency increased, and RDP N intake and predicted urine N output decreased with decreased concentration of dietary CP and RDP in the diets ( < 0.01). Blood metabolites were not affected by treatments. In conclusion, to improve the efficiency of N utilization by early-lactation dairy cows, 9.3% RDP in rations provides adequate protein to optimize milk

  14. Effects of parity and supply of rumen-degraded and undegraded protein on production and nitrogen balance in Holsteins.

    PubMed

    Flis, S A; Wattiaux, M A

    2005-06-01

    Eight Holstein cows (4 primiparous and 4 multiparous) were used in a replicated 4 x 4 Latin square design to determine milk production response and N balance when diets had no NRC-predicted excess of rumen-undegradable protein (RUP) or rumen-degradable protein (RDP), 10% RUP excess, 10% RDP excess, or 10% excess of both RUP and RDP. Diets were fed as a total mixed ration with (dry matter basis) 25% alfalfa silage, 25% corn silage, 19 to 21% corn grain, and varying proportions of solvent soybean meal and expeller soybean meal as primary sources of supplemental RDP and RUP, respectively. Milk yield and dry matter intake (DMI) were recorded daily, and total collection of feces and urine was completed in the last 3 d of each 21-d period. Dietary crude protein averaged 17.5 and 18.5% for the recommended and excess RDP diets, respectively, and 17.3 and 18.4% for the recommended and excess RUP diets, respectively. When cows were fed excess RUP diets in the form of expeller soybean meal, DMI and milk production increased, but the opposite was true when the diets contained excess RDP in the form of solvent soybean meal. Milk composition was not affected by RDP, RUP, or by parity, and there were no parity x RDP interactions for any of the measurements. However, apparent digestibility of neutral detergent fiber, dry matter, and N increased in multiparous cows but not in primiparous cows because of excess RUP. The increase in the yield of milk N with excess RUP was not influenced by parity, but multiparous cows retained more of the additional N apparently absorbed, whereas primiparous cows excreted the additional apparently absorbed N in the urine. Overall, the difference in urinary N due to parity (70 g/d) was about 4 times greater than the impact of dietary treatments (17 g/d). Our results suggest that multiparous cows have either a much larger urea pool or a greater demand to restore body protein mobilized earlier in lactation compared with primiparous cows. Reduction in

  15. Milk production and plasma gossypol of cows fed cottonseed and oilseed meals with or without rumen-undegradable protein.

    PubMed

    Blackwelder, J T; Hopkins, B A; Diaz, D E; Whitlow, L W; Brownie, C

    1998-11-01

    Twenty-four multiparous Holstein cows were randomly assigned at calving to treatment diets using a modified split-plot design to determine the effects of protein source on milk production and composition. The treatment diets consisted of an 80:20 combination of corn and alfalfa silages and whole cottonseed at 12% of the dietary dry matter (DM). The treatment diets were formulated to contain 17% crude protein (CP) and 20% acid detergent fiber on a DM basis. One of the following sources of supplemental CP was included in each treatment diet: 1) cottonseed meal, 2) cottonseed meal plus a rumen-undegradable protein (RUP) supplement, 3) soybean meal, and 4) soybean meal plus an RUP supplement. Cows were fed the initial treatment diet for 6 wk and then were switched to the other oilseed meal source but continued to receive the same amount of RUP during the second period of the study. Milk production and composition were not affected by treatment diet. Cows fed treatment diets without RUP supplementation consumed more DM and thus more CP. Supplementation with RUP resulted in greater milk production efficiency per unit of DM consumed. Cows fed treatment diets containing cottonseed meal had higher plasma gossypol concentrations than did cows fed treatment diets containing soybean meal. Plasma gossypol concentrations for all cows in each group were below the recommended upper limit that is considered to be safe. Data suggest that cottonseed meal in the diet can be substituted for soybean meal, resulting in similar milk production and composition.

  16. A simplified management of the in situ evaluation of feedstuffs in ruminants: Application to the study of the digestive availability of protein and amino acids corrected for the ruminal microbial contamination.

    PubMed

    González, Javier; Ouarti, Mafhoud; Rodríguez, Carlos Alberto; Centeno, Carmen

    2009-01-01

    The ruminal effective degradability (RED) and intestinal effective digestibility (IED) for dry matter, crude protein (CP) and amino acids (AA) were estimated by a simplified in situ method using pooled samples from rumen-incubated residues, which represented the ruminal outflow of undegraded feed. The effect of microbial contamination in the rumen was corrected using (15)N infusion techniques. Studies were carried out for soybean meal (SBM), barley grain (BG) and lucerne hay (LH) in three wethers cannulated in the rumen and the duodenum. Uncorrected values of RED for CP obtained either by mathematical integration or our simplified method were similar in all feeds. Microbial N in the pooled samples of SBM, BG and LH were 2%, 11% and 24% of total N, respectively. However, intestinal incubation eliminated this microbial charge by 100%, 99% and 88%, respectively. With microbial corrections, RED showed an increase, and IED showed a decrease, except for SBM. With this correction, intestinal digested CP was reduced by 2% in SBM, 13% in BG and 34% in LH. Corrected IED of AA was relatively similar in SBM (97-99%). However, large variations were observed in BG (74-93%) and in LH (10-88%). Digestion in the rumen and intestine changed the essential AA pattern. Overall, our results support that AA digestion is affected by the characteristics of their radicals and their contents in plant cell wall proteins. The accurate estimation of feed metabolisable AA or protein requires effective measures that are corrected by ruminal microbial contamination. The proposed in situ method largely simplifies these tasks and allows a more complete and less expensive feed evaluation.

  17. Abundance and Diversity of Dockerin-Containing Proteins in the Fiber-Degrading Rumen Bacterium, Ruminococcus flavefaciens FD-1

    PubMed Central

    Flint, Harry J.; Travis, Anthony J.; Jindou, Sadanari; Borovok, Ilya; Lamed, Raphael; Bayer, Edward A.; Henrissat, Bernard; Coutinho, Pedro M.; Antonopoulos, Dion A.; Berg Miller, Margret E.; White, Bryan A.

    2010-01-01

    protein, in response to their role in the rumen environment. PMID:20814577

  18. Effect of progressive inoculation of fauna-free sheep with holotrich protozoa and total-fauna on rumen fermentation, microbial diversity and methane emissions.

    PubMed

    Belanche, Alejandro; de la Fuente, Gabriel; Newbold, Charles J

    2015-03-01

    Rumen methanogenesis represents an energy waste for the ruminant and an important source of greenhouse gas; thus, integrated studies are needed to fully understand this process. Eight fauna-free sheep were used to investigate the effect of successive inoculation with holotrich protozoa then with total fauna on rumen methanogenesis. Holotrichs inoculation neither altered rumen fermentation rate nor diet digestibility, but increased concentrations of acetate (+15%), butyrate (+57%), anaerobic fungi (+0.82 log), methanogens (+0.41 log) and methanogenesis (+54%). Further inoculation with total fauna increased rumen concentrations of protozoa (+1.0 log), bacteria (+0.29 log), anaerobic fungi (+0.78 log), VFA (+8%), ammonia and fibre digestibility (+17%) without affecting levels of methanogens or methanogenesis. Rumen methanogens population was fairly stable in terms of structure and diversity, while the bacterial community was highly affected by the treatments. Inoculation with holotrich protozoa increased bacterial diversity. Further inoculation with total fauna lowered bacterial diversity but increased concentrations of certain propionate and lactate-producing bacteria, suggesting that alternative H2 sinks could be relevant. This experiment suggests that holotrich protozoa have a greater impact on rumen methanogenesis than entodiniomorphids. Thus, further research is warranted to understand the effect of holotrich protozoa on methane formation and evaluate their elimination from the rumen as a potential methane mitigation strategy. © Federation of European Microbiological Society 2014.

  19. Effect of progressive inoculation of fauna-free sheep with holotrich protozoa and total-fauna on rumen fermentation, microbial diversity and methane emissions

    PubMed Central

    Belanche, Alejandro; de la Fuente, Gabriel; Newbold, Charles J.

    2014-01-01

    Rumen methanogenesis represents an energy waste for the ruminant and an important source of greenhouse gas; thus, integrated studies are needed to fully understand this process. Eight fauna-free sheep were used to investigate the effect of successive inoculation with holotrich protozoa then with total fauna on rumen methanogenesis. Holotrichs inoculation neither altered rumen fermentation rate nor diet digestibility, but increased concentrations of acetate (+15%), butyrate (+57%), anaerobic fungi (+0.82 log), methanogens (+0.41 log) and methanogenesis (+54%). Further inoculation with total fauna increased rumen concentrations of protozoa (+1.0 log), bacteria (+0.29 log), anaerobic fungi (+0.78 log), VFA (+8%), ammonia and fibre digestibility (+17%) without affecting levels of methanogens or methanogenesis. Rumen methanogens population was fairly stable in terms of structure and diversity, while the bacterial community was highly affected by the treatments. Inoculation with holotrich protozoa increased bacterial diversity. Further inoculation with total fauna lowered bacterial diversity but increased concentrations of certain propionate and lactate-producing bacteria, suggesting that alternative H2 sinks could be relevant. This experiment suggests that holotrich protozoa have a greater impact on rumen methanogenesis than entodiniomorphids. Thus, further research is warranted to understand the effect of holotrich protozoa on methane formation and evaluate their elimination from the rumen as a potential methane mitigation strategy. PMID:25764558

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

    SciTech Connect

    Yu,P.

    2007-01-01

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

  1. Effect of supplementation of mustard oil cake on intake, digestibility and microbial protein synthesis of cattle in a straw-based diet in Bangladesh.

    PubMed

    Khandaker, Zahirul Haque; Uddin, Mohammad Mohi; Sultana, Nadira; Peters, Kurt J

    2012-04-01

    The objective of this study was to analyse the effects of different levels of rumen-degradable protein (RDP) on intake, digestibility and microbial protein synthesis by supplementing mustard oil cake (MOC) on rice straw-based diet of cattle (Bos indicus) in Bangladesh. A 4 × 4 Latin square design was applied. Four diets having constant energy (7.0 MJ/kg of dry matter (DM)) with varying levels of RDP (M(0) = 4.1 g/MJ (control), M(1) = 6.3 g/MJ, M(2) = 8.3 g/MJ and M(3) = 12.4 g/MJ of metabolizable energy (ME)) were received by each animal for a period of 28 days. A metabolism trial was conducted for 7 days. Results indicate that with increasing levels of RDP, crude protein (CP) and RDP intake increased significantly (P < 0.01). The significant (P < 0.01) increase in digestibility values are obtained for DM, organic matter, CP and digestible organic matter in the rumen. The digestibility of neutral detergent fibre and acid detergent fibre was also increased significantly (P < 0.05). The total nitrogen (N), ammonia-N and total volatile fatty acids increase significantly (P < 0.01) while the rumen pH increased from M(0) to M(2) and decreased thereafter. The efficiency microbial N intake increased significantly (P < 0.01) but showed a curvilinear response with higher RDP level (12.40 g/RDP/MJ ME). This study concludes that supplementation of RDP from MOC enhances the intake, digestibility and microbial protein synthesis which ultimately increases utilization of low-quality feed resources that can be used for developing cost-effective feeding systems on a straw-based diet in tropical regions.

  2. Effect of precision feeding high- and low-quality forage with different rumen protein degradability levels on nutrient utilization by dairy heifers.

    PubMed

    Ding, L M; Lascano, G J; Heinrichs, A J

    2015-06-01

    One approach to improving efficiency of protein and nitrogen use for optimal productivity in dairy heifers is to provide adequate levels of protein with differing rumen degradabilities. The objective of this experiment was to determine effects of manipulating RDP in diets of dairy heifers precision fed low- and high-quality forage. Eight rumen-cannulated Holstein heifers (448 ± 19 kg BW) were randomly used and assigned to an 85% forage diet with 2 forage qualities, low quality (LQ, 48% NDF) and high quality (HQ, 38% NDF), and to 4 RDP levels (62%, 68%, 74%, and 81% of CP as RDP) within forage quality administered according to a split-plot two 4 × 4 Latin square design (20-d periods). Similar N intake was provided (1.70 g N/kg BW0.75), and alterations of RDP levels were made by exchanging canola meal with heat-treated soy protein with slow-release urea. Heifers offered the HQ diet had greater apparent total tract DM and OM digestibilities than those fed the LQ diet. Apparent N digestibility tended to be higher for the HQ diet but was not affected by the levels of RDP in diets. No difference was observed for rumen VFA concentration between forage treatments, but acetate to propionate ratio was higher for LQ-fed heifers. No difference in rumen carbohydrate fermentation was detected with increasing levels of RDP. Feeding HQ forage improved DM, OM, and N apparent digestibility. Increasing RDP in LQ and HQ had no effects on apparent digestibility of any measured parameter or N dynamics in dairy heifers.

  3. Postprandial variations in the activity of polysaccharide-degrading enzymes in microbial populations from the digesta solids and liquor fractions of rumen contents.

    PubMed

    Williams, A G; Withers, S E; Strachan, N H

    1989-01-01

    The diurnal variations in the specific activities of polysaccharide-degrading enzymes after feeding were monitored in adherent and non-adherent microbial populations separated from bovine rumen liquor and digesta solids. There were marked differences in the activity profiles of the enzymes within the subpopulations. Enzymes involved in the degradation of soluble carbohydrates were more active in the non-adherent populations, and in the liquor phase subpopulation activities increased in the 1-2 h post-feed period. The muralytic enzymes were most active in the adherent population. Specific activities increased by up to 20-fold over the 24 h period, with an initial five-fold increase occurring between 8 h and 12 h after feeding. Enzyme levels in the three non-adherent populations were similar at the end of the postprandial period. In the population recovered from the liquid associated with the digesta particles, however, the activities did not increase until the latter stages of the period, whereas in the non-adherent population from the digesta solids the activities varied little during the diurnal cycle. The numbers of micro-organisms associated with the digesta solids were similar at 2 h and 20 h after feeding; the variations in enzyme levels did not occur as a result of a population increase but were due to increased activities in an established population. The plant cell wall structural polysaccharides were degraded at different rates. There was no appreciable cellulose digestion during the first 8 h of the postprandial period and although hemicellulosic constituents were removed continuously the rate of loss of both polymers was increased in the later stages of the diurnal cycle when enzyme activities were maximal.

  4. Diversity of rumen bacteria in canadian cervids.

    PubMed

    Gruninger, Robert J; Sensen, Christoph W; McAllister, Timothy A; Forster, Robert J

    2014-01-01

    Interest in the bacteria responsible for the breakdown of lignocellulosic feedstuffs within the rumen has increased due to their potential utility in industrial applications. To date, most studies have focused on bacteria from domesticated ruminants. We have expanded the knowledge of the microbial ecology of ruminants by examining the bacterial populations found in the rumen of non-domesticated ruminants found in Canada. Next-generation sequencing of 16S rDNA was employed to characterize the liquid and solid-associated bacterial communities in the rumen of elk (Cervus canadensis), and white tailed deer (Odocoileus virginianus). Despite variability in the microbial populations between animals, principle component and weighted UniFrac analysis indicated that bacterial communities in the rumen of elk and white tail deer are distinct. Populations clustered according to individual host animal and not the association with liquid or solid phase of the rumen contents. In all instances, Bacteroidetes and Firmicutes were the dominant bacterial phyla, although the relative abundance of these differed among ruminant species and between phases of rumen digesta, respectively. In the elk samples Bacteroidetes were more predominant in the liquid phase whereas Firmicutes was the most prevalent phyla in the solid digesta (P = 1×10(-5)). There were also statistically significant differences in the abundance of OTUs classified as Fibrobacteres (P = 5×10(-3)) and Spirochaetes (P = 3×10(-4)) in the solid digesta of the elk samples. We identified a number of OTUs that were classified as phylotypes not previously observed in the rumen environment. Our results suggest that although the bacterial diversity in wild North American ruminants shows overall similarities to domesticated ruminants, we observed a number of OTUs not previously described. Previous studies primarily focusing on domesticated ruminants do not fully represent the microbial diversity of the rumen and

  5. Diversity of Rumen Bacteria in Canadian Cervids

    PubMed Central

    Gruninger, Robert J.; Sensen, Christoph W.; McAllister, Timothy A.; Forster, Robert J.

    2014-01-01

    Interest in the bacteria responsible for the breakdown of lignocellulosic feedstuffs within the rumen has increased due to their potential utility in industrial applications. To date, most studies have focused on bacteria from domesticated ruminants. We have expanded the knowledge of the microbial ecology of ruminants by examining the bacterial populations found in the rumen of non-domesticated ruminants found in Canada. Next-generation sequencing of 16S rDNA was employed to characterize the liquid and solid-associated bacterial communities in the rumen of elk (Cervus canadensis), and white tailed deer (Odocoileus virginianus). Despite variability in the microbial populations between animals, principle component and weighted UniFrac analysis indicated that bacterial communities in the rumen of elk and white tail deer are distinct. Populations clustered according to individual host animal and not the association with liquid or solid phase of the rumen contents. In all instances, Bacteroidetes and Firmicutes were the dominant bacterial phyla, although the relative abundance of these differed among ruminant species and between phases of rumen digesta, respectively. In the elk samples Bacteroidetes were more predominant in the liquid phase whereas Firmicutes was the most prevalent phyla in the solid digesta (P = 1×10−5). There were also statistically significant differences in the abundance of OTUs classified as Fibrobacteres (P = 5×10−3) and Spirochaetes (P = 3×10−4) in the solid digesta of the elk samples. We identified a number of OTUs that were classified as phylotypes not previously observed in the rumen environment. Our results suggest that although the bacterial diversity in wild North American ruminants shows overall similarities to domesticated ruminants, we observed a number of OTUs not previously described. Previous studies primarily focusing on domesticated ruminants do not fully represent the microbial diversity of the rumen and

  6. Microbial protein production: maximizing protein production efficiency in Space habitats

    NASA Astrophysics Data System (ADS)

    Clauwaert, Peter; Alloul, Abbas; Muys, Maarten; Sui, Yixing; Boon, Nico; Luther, Amanda; Christiaens, Marlies E. R.; Ilgrande, Chiara; Lindeboom, Ralph E. F.; Rabaey, Korneel; Vlaeminck, Siegfried

    2016-07-01

    On top of the goal of a closed material cycle for Space habitats or deep Space missions with food production, extreme requirements apply to such Life Support Systems (LSS) in terms of mass, volume, crew time, energy consumption and controllability. Although relatively high water recovery efficiencies (~70-90%) can be achieved, all Space missions until now have relied on terrestrial food resupply and thus no nutrient recovery has been achieved so far. Researchers and Space agencies have typically been focussing on the cultivation of higher plants to produce food for crew members for future Space LSS. It can be assumed that the required surface area (50-500 m2 per crew member), plant evaporation rates (~200 kg per crew member per day), power consumption (~65 kW per crew member) and the degree of controllability of a higher plant compartment will have a great impact on the feasibility of realizing a future closed loop LSS in Space for the first time. As the food production density is so critical in a LSS, a combination of higher plant cultivation and microbial protein production might increase the chances of success of future Space LSS's since the production densities are significantly higher. Higher plants in Space LSS's would typically have an average specific protein production rate in the order of 0-4 kg protein m-3 year-1 (calculated from Do, Owens et al. (2016)), whereas bacterial biomass can be produced continuously at a rate up to ~1000 kg protein m-3 year-1. Several routes for microbial food production will be discussed in this presentation, ranging from aerobic heterotrophic production with for instance Candida ingens (Strayer, Finger et al. 1997), photoheterotrophic production with PNSB such as Rhodospirillum rubrum (Hendrickx, De Wever et al. 2006) and hydrogenotrophic production with HOB such as Cupriavidus necator (Matassa, Boon et al. 2015)) and photoautotrophic production of oxygen and microbial food (e.g. Arthrospira sp. (Hendrickx, De Wever et al

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

    PubMed

    Patra, Amlan K; Saxena, Jyotisna

    2011-01-15

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

  8. Effects of nitrate addition to a diet on fermentation and microbial populations in the rumen of goats, with special reference to Selenomonas ruminantium having the ability to reduce nitrate and nitrite.

    PubMed

    Asanuma, Narito; Yokoyama, Shota; Hino, Tsuneo

    2015-04-01

    This study investigated the effects of dietary nitrate addition on ruminal fermentation characteristics and microbial populations in goats. The involvement of Selenomonas ruminantium in nitrate and nitrite reduction in the rumen was also examined. As the result of nitrate feeding, the total concentration of ruminal volatile fatty acids decreased, whereas the acetate : propionate ratio and the concentrations of ammonia and lactate increased. Populations of methanogens, protozoa and fungi, as estimated by real-time PCR, were greatly decreased as a result of nitrate inclusion in the diet. There was modest or little impact of nitrate on the populations of prevailing species or genus of bacteria in the rumen, whereas Streptococcus bovis and S. ruminantium significantly increased. Both the activities of nitrate reductase (NaR) and nitrite reductase (NiR) per total mass of ruminal bacteria were increased by nitrate feeding. Quantification of the genes encoding NaR and NiR by real-time PCR with primers specific for S. ruminantium showed that these genes were increased by feeding nitrate, suggesting that the growth of nitrate- and nitrite-reducing S. ruminantium is stimulated by nitrate addition. Thus, S. ruminantium is likely to play a major role in nitrate and nitrite reduction in the rumen. © 2014 Japanese Society of Animal Science.

  9. Effects of BioChlor and Fermenten on microbial protein synthesis in continuous culture fermenters.

    PubMed

    Lean, I J; Webster, T K Miller; Hoover, W; Chalupa, W; Sniffen, C J; Evans, E; Block, E; Rabiee, A R

    2005-07-01

    Meta analysis models were constructed from a data-set of 15 continuous culture fermenter trials and 118 observations on studies with either BioChlor (n = 23 observations) or Fermenten (n = 95) included at 10 and 3%, respectively, of dietary dry matter (DM) to evaluate effects of the ingredients BioChlor and Fermenten (B/F) on rumen function. Digestibility of crude protein was significantly increased by 11% with B/F treatment. This was reflected in significant increases in digestibility of DM and organic matter (OM) by 3.6 and 7.9%, respectively. Increased amounts of sugar in the diet in the presence of B/F tended to reduce digestibility of non-structural carbohydrates (NSC); however, the net effect on NSC digestion was small. There was no effect of treatment on most individual volatile fatty acids (VFA) or total VFA production. Propionate production, however, was significantly reduced in treated fermenters. The main effect of B/F as well as of starch and soluble fiber when combined with the treatment was to increase propionate production; however, the interaction between B/F treatment and sugar decreased propionate production markedly, resulting in a net decrease. The acetate-to-propionate ratio increased by 6% with B/F, largely as a result of the decrease in propionate. Production of nonammonia nitrogen was 1% less in B/F-treated fermenters, and interactions between treatment and starch, sugar, or soluble fiber were significant. Treated fermenters produced 15.7% more microbial nitrogen, in association with a significant 37% increase in rumen protein digestion. Interactions between treatment and starch, soluble fiber, or sugar influenced these results. The interaction of B/F and sugar resulted in a decrease in undegradable protein N and an increase in microbial nitrogen production. Ammonia nitrogen concentrations were increased by 24.6% in treated fermenters. Efficiency of microbial nitrogen production from DM, OM, or carbohydrate was significantly increased by B

  10. Novel technologies in rumen microbiology: What have we learned?

    USDA-ARS?s Scientific Manuscript database

    Beginning in the 1950s, rumen microbiology enjoyed a golden age in which the secrets of the rumen began to be teased out through the isolation of numerous new microbial species and the discovery of such important concepts as interspecies hydrogen transfer and the quantitative aspects of bacterial gr...

  11. Characterization of the rumen microbiome of Indian Kankrej cattle (Bos indicus) adapted to different forage diet.

    PubMed

    Patel, Vilas; Patel, Amrutlal K; Parmar, Nidhi R; Patel, Anand B; Reddy, Bhaskar; Joshi, Chaitanya G

    2014-12-01

    Present study described rumen microbiome of Indian cattle (Kankrej breed) to better understand the microbial diversity and largely unknown functional capacity of the rumen microbiome under different dietary treatments. Kankrej cattle were gradually adapted to a high-forage diet (four animals with dry forage and four with green forage) containing 50 % (K1), 75 % (K2) to 100 % (K3) forage, and remaining concentrate diet, each for 6 weeks followed by analysis of rumen fiber adherent and fiber-free metagenomic community by shotgun sequencing using ion torrent PGM platform and EBI-metagenomics annotation pipeline. Taxonomic analysis indicated that rumen microbiome was dominated by Bacteroidetes followed by Firmicutes, Fibrobacter, Proteobacteria, and Tenericutes. Functional analysis based on gene ontology classified all reads in total 157 categories based on their functional role in biological, molecular, and cellular component with abundance of genes associated with hydrolase activity, membrane, transport, transferase, and different metabolism (such as carbohydrate and protein). Statistical analysis using STAMP revealed significant differences (P < 0.05) between solid and liquid fraction of rumen (in 65 categories), between all three treatments (in 56 categories), and between green and dry roughage (17 categories). Diet treatment also exerted significant difference in environmental gene tags (EGTs) involved in metabolic pathways for production of volatile fatty acids. EGTs for butyrate production were abundant in K2, whereas EGTs for propionate production was abundant during K1. Principal component analysis also demonstrated that diet proportion, fraction of rumen, and type of forage affected rumen microbiome at taxonomic as well as functional level.

  12. Energy utilization, nitrogen balance and microbial protein supply in cattle fed Pennisetum purpureum and condensed tannins.

    PubMed

    Piñeiro-Vázquez, A T; Canul-Solis, J R; Alayón-Gamboa, J A; Chay-Canul, A J; Ayala-Burgos, A J; Solorio-Sánchez, F J; Aguilar-Pérez, C F; Ku-Vera, J C

    2017-02-01

    The aim of the experiment was to assess the effect of condensed tannins (CT) on feed intake, dry matter digestibility, nitrogen balance, supply of microbial protein to the small intestine and energy utilization in cattle fed a basal ration of Pennisetum purpureum grass. Five heifers (Bos taurus × Bos indicus) with an average live weight of 295 ± 19 kg were allotted to five treatments consisting of increasing levels of CT (0, 1, 2, 3 and 4% CT/kg DM) in a 5 × 5 Latin square design. Dry matter intake (DMI) was similar (p > 0.05) between treatments containing 0, 1, 2 and 3% of CT/kg DM and it was reduced (p < 0.05) to 4% CT (5.71 kg DM/day) with respect to that observed with 0% CT (6.65 kg DM/day). Nitrogen balance, purine derivatives excretion in urine, microbial protein synthesis and efficiency of synthesis of microbial nitrogen in the rumen were not affected (p ≥ 0.05) by the increase in the levels of condensed tannins in the ration. Energy loss as CH4 was on average 2.7% of the gross energy consumed daily. Metabolizable energy intake was 49.06 MJ/day in cattle fed low-quality tropical grass with a DMI of 6.27 kg/day. It is concluded that concentrations of CT between 2 and 3% of DM of ration reduced energy loss as CH4 by 31.3% and 47.6%, respectively, without affecting intakes of dry and organic matter; however, digestibilities of dry and organic matter are negatively affected. Journal of Animal Physiology and Animal Nutrition © 2016 Blackwell Verlag GmbH.

  13. The Escherichia coli O157:H7 bovine rumen fluid proteome reflects adaptive bacterial responses.

    PubMed

    Kudva, Indira T; Stanton, Thaddeus B; Lippolis, John D

    2014-02-21

    To obtain insights into Escherichia coli O157:H7 (O157) survival mechanisms in the bovine rumen, we defined the growth characteristics and proteome of O157 cultured in rumen fluid (RF; pH 6.0-7.2 and low volatile fatty acid content) obtained from rumen-fistulated cattle fed low protein content "maintenance diet" under diverse in vitro conditions. Bottom-up proteomics (LC-MS/MS) of whole cell-lysates of O157 cultured under anaerobic conditions in filter-sterilized RF (fRF; devoid of normal ruminal microbiota) and nutrient-depleted and filtered RF (dRF) resulted in an anaerobic O157 fRF-and dRF-proteome comprising 35 proteins functionally associated with cell structure, motility, transport, metabolism and regulation, but interestingly, not with O157 virulence. Shotgun proteomics-based analysis using isobaric tags for relative and absolute quantitation used to further study differential protein expression in unfiltered RF (uRF; RF containing normal rumen microbial flora) complemented these results. Our results indicate that in the rumen, the first anatomical compartment encountered by this human pathogen within the cattle gastrointestinal tract (GIT), O157 initiates a program of specific gene expression that enables it to adapt to the in vivo environment, and successfully transit to its colonization sites in the bovine GIT. Further experiments in vitro using uRF from animals fed different diets and with additional O157 strains, and in vivo using rumen-fistulated cattle will provide a comprehensive understanding of the adaptive mechanisms involved, and help direct evolution of novel modalities for blocking O157 infection of cattle.

  14. The Escherichia coli O157:H7 bovine rumen fluid proteome reflects adaptive bacterial responses

    PubMed Central

    2014-01-01

    Background To obtain insights into Escherichia coli O157:H7 (O157) survival mechanisms in the bovine rumen, we defined the growth characteristics and proteome of O157 cultured in rumen fluid (RF; pH 6.0-7.2 and low volatile fatty acid content) obtained from rumen-fistulated cattle fed low protein content “maintenance diet” under diverse in vitro conditions. Results Bottom-up proteomics (LC-MS/MS) of whole cell-lysates of O157 cultured under anaerobic conditions in filter-sterilized RF (fRF; devoid of normal ruminal microbiota) and nutrient-depleted and filtered RF (dRF) resulted in an anaerobic O157 fRF-and dRF-proteome comprising 35 proteins functionally associated with cell structure, motility, transport, metabolism and regulation, but interestingly, not with O157 virulence. Shotgun proteomics-based analysis using isobaric tags for relative and absolute quantitation used to further study differential protein expression in unfiltered RF (uRF; RF containing normal rumen microbial flora) complemented these results. Conclusions Our results indicate that in the rumen, the first anatomical compartment encountered by this human pathogen within the cattle gastrointestinal tract (GIT), O157 initiates a program of specific gene expression that enables it to adapt to the in vivo environment, and successfully transit to its colonization sites in the bovine GIT. Further experiments in vitro using uRF from animals fed different diets and with additional O157 strains, and in vivo using rumen-fistulated cattle will provide a comprehensive understanding of the adaptive mechanisms involved, and help direct evolution of novel modalities for blocking O157 infection of cattle. PMID:24559513

  15. Molecular basis of protein structure in combined feeds (hulless barley with bioethanol coproduct of wheat dried distillers grains with solubles) in relation to protein rumen degradation kinetics and intestinal availability in dairy cattle.

    PubMed

    Zhang, X; Yu, P

    2012-06-01

    The objectives of this study were to reveal protein molecular structure in relation to rumen degradation kinetics and intestinal availability in combined feeds of hulless barley with bioethanol coproduct [pure wheat dried distillers grains with solubles (DDGS)] at 5 different ratios (100:0, 75:25, 50:50, 25:75, and 0:100) in dairy cattle. The parameters assessed included 1) protein chemical profiles, 2) protein subfractions partitioned by the Cornell Net Carbohydrate and Protein System, 3) in situ protein degradation kinetics, 4) truly absorbed protein supply in the small intestine (DVE), metabolizable protein characteristics and degraded protein balance (OEB), 5) protein molecular structure spectral profiles, and 6) correlation between protein molecular structure and protein nutrient profiles and metabolic characteristics. We found that 1) with increasing inclusion of wheat DDGS in feed combinations, protein chemical compositions of crude protein (CP), neutral detergent-insoluble CP, acid detergent-insoluble CP, and nonprotein N were increased, whereas soluble CP was decreased linearly; CP subfractions A, B₃, and C were increased linearly, but CP subfractions B₁ and B₂ were decreased; truly digestible CP increased but total digestible nutrients at 1× maintenance decreased linearly; protein degradation rate was decreased without affecting potentially soluble, potentially degradable, and potentially undegradable fractions, and both rumen-degradable protein and rumen-undegradable protein were increased; by using the DVE/OEB system, the DVE and OEB values were increased from 98 to 226 g/kg of dry matter and -1 to 105 g/kg of dry matter, respectively; 2) by using the molecular spectroscopy technique, the spectral differences in protein molecular structure were detected among the feed combinations; in the original combined feeds, amide I and II peak area and ratio of amide I to II were increased linearly; although no difference existed in α-helix and

  16. Effects of replacing soybean meal with canola meal differing in rumen-undegradable protein content on ruminal fermentation and gas production kinetics using 2 in vitro systems.

    PubMed

    Paula, E M; Monteiro, H F; Silva, L G; Benedeti, P D B; Daniel, J L P; Shenkoru, T; Broderick, G A; Faciola, A P

    2017-07-01

    Previous research indicated that there were significant differences in rumen-undegradable protein (RUP) among canola meals (CM), which could influence the nutritional value of CM. The objectives of this study were to (1) evaluate the effects of feeding CM with different RUP contents on ruminal fermentation, nutrient digestion, and microbial growth using a dual-flow continuous culture system (experiment 1) and (2) evaluate ruminal gas production kinetics, in vitro organic matter (OM) digestibility, and methane (CH4) production of soybean meal (SBM) and CM with low or high RUP in the diet or as a sole ingredient using a gas production system (experiments 2 and 3). In experiment 1, diets were randomly assigned to 6 fermentors in a replicated 3 × 3 Latin square. The only ingredient that differed among diets was the protein supplement. The treatments were (1) solvent-extracted SBM, (2) low-RUP solvent-extracted CM (38% RUP as a percentage of crude protein), and (3) high-RUP solvent-extracted CM (50% RUP). Diets were prepared as 3 concentrate mixtures that were combined with 25% orchardgrass hay and 15% wheat straw (dry matter basis). Experiments 2 and 3 had the same design with 24 bottles incubated 3 times for 48 h each. During the 48-h incubation, the cumulative pressure was recorded to determine gas production kinetics, in vitro OM digestibility, and CH4 production. In experiment 1, N flow (g/d), efficiency of N use, efficiency of bacterial N synthesis, total volatile fatty acids (mM), and molar proportion of acetate, propionate, and isobutyrate were not affected by treatments. There were tendencies for a decrease in ruminal NH3-N and an increase in molar proportion of butyrate for the SBM diet compared with both CM diets. The molar proportion of valerate was greater in both CM diets, whereas the molar proportion of isovalerate and total branched-chain volatile fatty acids was lower for the CM diets compared with the SBM diet. In experiments 2 and 3, the SBM diet had

  17. Replacement of alfalfa hay (Medicago sativa L.) with subabul (Leucaena leucocephala) leaf meal in diets of Najdi goats: effect on digestion activity of rumen microorganisms.

    PubMed

    Mohammadabadi, Tahereh; Jolazadeh, Alireza

    2017-08-01

    This study investigated the effect of replacing alfalfa hay by subabul leaf meal (SLM) on digestion, fermentation parameters and rumen bacteria and fungi activity of Najdi goats. Six Najdi goats (150 ± 15 days of age and initial body weight of 35 ± 1.1 kg) were randomly assigned to one of two dietary treatments in a balanced completely randomized design (three goats per treatment) for 56 days. Experimental treatments included alfalfa hay as control group and diet containing SLM (SLM replacing alfalfa hay at 50% level). Bacterial and fungi activity and rumen fermentation parameters of animals fed experimental diets were determined. Dry matter disappearance (DMD) was unaffected by replacing SLM with alfalfa hay for both rumen bacteria and fungi in different incubation times, except for 48 h of incubation in specific culture medium of mixed rumen bacteria, which decreased for SLM group (P > 0.05). NDF disappearance (NDFD) and ADF disappearance (ADFD) after 24 and 48 h of incubation in specific culture medium of mixed rumen bacteria was not affected by experimental diets (P > 0.05). However, 72 h after incubation, NDFD in SLM treatment decreased (P > 0.05). Gas production parameters of rumen bacteria were similar for both experimental diets, but partitioning factor (PF), efficiency microbial biomass production (EMBP), microbial protein production (MP), and organic matter truly digested (OMTD) decreased (p < 0.05) in specific culture medium of mixed rumen fungi for SLM diet. There was significant decrease in total protozoa population for diet containing SLM (p < 0.05). Results indicated that replacement of alfalfa hay by SLM had no major effect on rumen microorganisms' activity of Najdi goats, so it may be used as an alternative for alfalfa (at 50% level) in susceptible areas.

  18. Comparison of grass and legume silages for milk production. 2. In vivo and in sacco evaluations of rumen function.

    PubMed

    Dewhurst, R J; Evans, R T; Scollan, N D; Moorby, J M; Merry, R J; Wilkins, R J

    2003-08-01

    Two experiments were conducted to investigate the basis for higher voluntary intakes and increased alpha-linolenic acid content in milk from cows offered clover silages. Six cows with rumen and duodenal cannulae were used in a four-period changeover-design experiment. Cows received 8 kg/d of dairy concentrate and had ad libitum access to one of six silage treatments: grass, red clover, white clover, alfalfa, and 50/50 (dry matter basis) mixtures of grass with red clover or white clover. The rumen fermentability of grass, red clover, white clover, and grass/red clover silages was also evaluated in a nylon bag study. Legume silages led to increased dry matter intake and milk production in comparison with grass silage. There was no significant effect of legume silages on rumen pH and volatile fatty acid concentrations, but a significant increase in rumen ammonia concentration with the legume silages, reflecting their higher protein content. The inclusion of white clover or alfalfa silage, but not red clover silage, in diets led to an increase in molar proportions of isobutyric, iso-valeric, and n-valeric acids in comparison with diets based on grass silage. Rumen fill was significantly lower, and rumen passage rates were significantly higher for cows offered alfalfa or white clover silages. However, the markedly different particle size distribution of rumen contents with these feeds suggests very different mechanisms for the high intake characteristics: high rates of particle breakdown and passage with alfalfa, and high rates of fermentation and passage with white clover. Microbial energetic efficiency (grams microbial N per kilogram organic matter apparently digested in the rumen) was highest for cows offered alfalfa silage, intermediate for clover silage, and lowest for cows offered grass silage. These differences reflect the higher rumen outflow rates for legume silages in comparison with grass silage. However, the effect of these differences on N-use efficiency

  19. Metabolic Fate of Cysteine and Methionine in Rumen Digesta

    PubMed Central

    Nader, C. J.; Walker, D. J.

    1970-01-01

    Estimates were obtained of the extent to which cysteine and methionine were incorporated into the protein of the microbes of rumen digesta without prior degradation and resynthesis. By using the amino acids labeled with both 35S and 14C, it was observed that a large proportion of the 35S appeared in the sulfide pool and of the 14C appeared in volatile fatty acids. By isolating the appropriate amino acid, obtaining the 14C to 35S ratio, and comparing this with the ratio in the added amino acid, the degree of direct incorporation was calculated. For cysteine it was estimated that at most 1% and for methionine, at most 11% of the amino acid in the free pool was incorporated unchanged into microbial protein. As a consequence of these findings, it is considered that the method for measuring microbial protein synthesis in rumen digesta based upon incorporation of 35S from the free sulfide pool is not seriously affected by direct utilization of sulfur amino acids arising from dietary sources. PMID:5485079

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

    PubMed

    Patra, Amlan K; Yu, Zhongtang

    2014-01-01

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

  1. Effect of bovine somatotropin and rumen-undegradable protein on mammary growth of prepubertal dairy heifers and subsequent milk production.

    PubMed

    Capuco, A V; Dahl, G E; Wood, D L; Moallem, U; Erdman, R E

    2004-11-01

    Rapid body growth during the prepubertal period may be associated with reductions in mammary parenchymal growth and subsequent milk yield. The objective of this study was to test effects of dietary rumen-undegradable protein (RUP) and administration of recombinant bovine somatotropin (bST) during the prepubertal period on mammary growth and milk yield of dairy heifers. Seventy-two Holstein heifers were used in the experiment. At 90 d of age, 8 heifers were slaughtered before initiation of treatment. Remaining heifers were assigned randomly to 1 of 4 treatments. Treatments consisted of a control diet (5.9% RUP, 14.9% CP, DM basis) or RUP-supplemented diet (control diet plus 2% added RUP) with or without 0.1 mg of bST/kg of BW per day applied in a 2 x 2 factorial design. A total of 6 heifers per treatment (3 each at 5 and 10 mo of age) were slaughtered for mammary tissue analysis. Remaining heifers were bred to evaluate impact of treatment on subsequent milk yield and composition. Mammary parenchymal growth was not affected by RUP or bST treatment. Total parenchymal mass increased from 16 to 364 g, and parenchymal DNA from 58 to 1022 mg from 3 to 10 mo of age, respectively. Furthermore, number of mammary epithelial cells likely was not affected by diet or bST because the epithelial cell proliferation index, assessed by Ki-67 labeling, was not affected by treatment, nor was total parenchymal DNA and lipid content. Neither deleterious effects of increased rates of gain nor positive effects of bST were evident in prepubertal mammary growth. Subsequent milk production and composition was not different among treatments.

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

    PubMed

    Ohlsson, C; Houmøller, L P; Weisbjerg, M R; Lund, P; Hvelplund, T

    2007-12-01

    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 was described in terms of immediately rumen soluble fraction (a fraction, for DM and CP only as NDF does not contain a soluble fraction), the degradable but not soluble faction (b fraction) and the rate of degradation of the b fraction (c value). Overall effective degradability of DM, CP and NDF was also calculated. Near infrared reflectance spectroscopy was examined for its ability to predict degradation parameters and to make a direct prediction of effective degradation from scans of the original samples of perennial ryegrass and orchardgrass. Prediction of effective degradation of the different feed fractions showed different accuracy. The coefficients of determination (R(2)) from regressions of predicted vs. measured effective degradation, using a cross-validation method, were 0.92 for DM, 0.78 for CP and 0.61 for NDF. The attempt to predict the degradation parameters (a, b and c) by NIRS was less successful as the coefficients of determination for the degradation parameters were low. Concentrations of CP and NDF in the original samples were predicted by using NIRS and the validated R(2) value was 0.98 for CP and 0.92 for NDF. It is concluded that using NIRS predictions from scans of original samples is a promising method to obtain values for the effective degradation of DM, CP and NDF in ruminant feeds, but that larger calibration sets are necessary for obtaining improved

  3. Effect of phytosterols on rumen fermentation in vitro.

    PubMed

    Xi, Y M; Jin, Z H; Lin, L J; Han, Z Y

    2014-05-16

    We investigated the effect of phytosterols on rumen fermentation in vitro using gas syringes as incubators. Phytosterols were dissolved in ethyl acetate (8.3%) and added at various concentrations to the common diet in rumen fluid. In vitro gas production (GP) was recorded after 3, 6, 12, 18, and 24 h incubation. Incubation was stopped at 6, 12, and 24 h and the inoculants were then tested for pH, dry matter digestibility (DMD), microbial protein yield (MCP), lactic acid, NH3-N, and volatile fatty acids (VFAs). GP was consistently higher than the control; particularly, treatments at 12, 18, and 24 h reached extremely significant levels (P < 0.01). Compared to the control group, the pH of ruminal fluid was slightly lower after incubation, and DMD and MCP increased with increasing phytosterol level except for the content of MCP at 6 h, which changed only minimally. Lactate was significantly lower after treatment compared to the control at 12 h (P < 0.01) and 24 h (P < 0.05), while NH3-N at 12 h (P < 0.05) and 24 h (P < 0.01) after treatment decreased significantly. Acetate, propionate, butyrate, and total VFA for all treatments were higher than those of the control, particularly for butyrate at 6 h (P < 0.01). These results suggest that phytosterols modify rumen fermentation by inhibiting released harmful products and promoting the release of beneficial product, which may be useful for improving nutrient utilization and animal health.

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

    PubMed

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

    2014-01-01

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

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

    PubMed Central

    Soliva, Carla R.; Amelchanka, Sergej L.; Kreuzer, Michael

    2015-01-01

    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 for 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 by

  6. Feeding rumen-protected methionine pre- and postpartum increases milk protein content and yield in early lactation

    USDA-ARS?s Scientific Manuscript database

    Objectives were to evaluate the effects of feeding rumen-protected methionine (MET) from 23 d (±12) before calving until 98 days in milk (DIM) on lactation performance, dry matter intake (DMI), body condition score (BCS) and body weight (BW) change of dairy cows. Multiparous Holstein cows (n = 223) ...

  7. Effect of incremental urea supplementation of a conventional corn silage-based diet on ruminal ammonia concentration and synthesis of microbial protein.

    PubMed

    Boucher, S E; Ordway, R S; Whitehouse, N L; Lundy, F P; Kononoff, P J; Schwab, C G

    2007-12-01

    One primiparous and 3 multiparous lactating Holstein cows fitted with ruminal and duodenal cannulas were used in a 4 x 4 Latin square design to determine the efficacy of adding urea to a corn silage-based diet on ruminal fermentation and microbial protein synthesis. Dietary treatments were 0, 0.3, 0.6, and 0.9% urea in diet dry matter (DM); urea was manually top dressed and incorporated into the ration. The basal diet contained (DM basis) 52% forage (with 61% of forage provided as corn silage) and 48% concentrate ingredients. The basal diet was formulated to meet National Research Council (NRC, 2001) requirements for energy and all nutrients except rumen-degradable protein (RDP) and metabolizable protein. Experimental periods lasted 14 d with the first 9 d for adaptation. The basal diet, without urea addition, contained 9.2% RDP in DM and had a predicted RDP balance of -167 g/d (NRC, 2001). There were no effects of dietary treatment on ruminal true digestibility of organic matter or ruminal apparent digestibility of neutral detergent fiber and acid detergent fiber. Total ruminal volatile fatty acid concentrations increased linearly with increasing urea level. Feeding increasing amounts of urea quadratically increased rumen ammonia N concentrations (9.0, 11.9, 12.8, and 17.4 mg/dL at 0, 0.3, 0.6, and 0.9% urea supplementation, respectively), passage of microbial N, and microbial N in duodenal digesta as a percentage of nonammonia N. The results of this study indicate that there were some positive effects of adding urea to the described lactating dairy cow diet, and that microbial protein synthesis was maximized at an average ruminal ammonia N concentration of 12.8 mg/dL when urea was added at 0.6% in diet DM.

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

    PubMed

    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

  9. Development of feeding systems and strategies of supplementation to enhance rumen fermentation and ruminant production in the tropics

    PubMed Central

    2013-01-01

    The availability of local feed resources in various seasons can contribute as essential sources of carbohydrate and protein which significantly impact rumen fermentation and the subsequent productivity of the ruminant. Recent developments, based on enriching protein in cassava chips, have yielded yeast fermented cassava chip protein (YEFECAP) providing up to 47.5% crude protein (CP), which can be used to replace soybean meal. The use of fodder trees has been developed through the process of pelleting; Leucaena leucocephala leaf pellets (LLP), mulberry leaf pellets (MUP) and mangosteen peel and/or garlic pellets, can be used as good sources of protein to supplement ruminant feeding. Apart from producing volatile fatty acids and microbial proteins, greenhouse gases such as methane are also produced in the rumen. Several methods have been used to reduce rumen methane. However, among many approaches, nutritional manipulation using feed formulation and feeding management, especially the use of plant extracts or plants containing secondary compounds (condensed tannins and saponins) and plant oils, has been reported. This approach could help todecrease rumen protozoa and methanogens and thus mitigate the production of methane. At present, more research concerning this burning issue - the role of livestock in global warming - warrants undertaking further research with regard to economic viability and practical feasibility. PMID:23981662

  10. Determining the culturability of the rumen bacterial microbiome

    PubMed Central

    Creevey, Christopher J; Kelly, William J; Henderson, Gemma; Leahy, Sinead C

    2014-01-01

    The goal of the Hungate1000 project is to generate a reference set of rumen microbial genome sequences. Toward this goal we have carried out a meta-analysis using information from culture collections, scientific literature, and the NCBI and RDP databases and linked this with a comparative study of several rumen 16S rRNA gene-based surveys. In this way we have attempted to capture a snapshot of rumen bacterial diversity to examine the culturable fraction of the rumen bacterial microbiome. Our analyses have revealed that for cultured rumen bacteria, there are many genera without a reference genome sequence. Our examination of culture-independent studies highlights that there are few novel but many uncultured taxa within the rumen bacterial microbiome. Taken together these results have allowed us to compile a list of cultured rumen isolates that are representative of abundant, novel and core bacterial species in the rumen. In addition, we have identified taxa, particularly within the phylum Bacteroidetes, where further cultivation efforts are clearly required. This information is being used to guide the isolation efforts and selection of bacteria from the rumen microbiota for sequencing through the Hungate1000. PMID:24986151

  11. Determination of the dry matter disappearance and the crude protein degradability of the formaldehyde treated feeds in the rumen by in situ method.

    PubMed

    Ceresnáková, Z; Chrenková, M; Sommer, A; Szakács, J

    1989-01-01

    Sunflower cake (SC), horse bean (HB), and alfalfa meal (AM) were treated with rising doses of formaldehyde (F) from 0.0, 0.2 ... 2.0 g F per 100 g crude protein. We have followed the differences of crude protein solubility, its degradability and disappearance of dry matter in the rumen as well as the amount of reversible bonded F from the F dose. With AM it was found a significant influence of ADF on the degradability and on the amount of acid-labile bonded F and/or on the decrease of N solubility. From the total N content the share of soluble nitrogen decreased in SC by 31%, HB by 24%, and in AM only by 12.5%. With the increase of the F dose its protective effect on proteins against the degradability mainly in SC and HB also rose significantly. The degradability of AM protein was influenced more by the incubation time than by the effect of treatment. The degradability of the SC proteins fell from 95% with untreated to 27% treated with the maximum dose. With AM it fell slowly from 84% to 74% by the determination coefficient R2 = 0.77. It was observed a marked decrease in HB only from the dose of 1.28% F. The changes in dry matter disappearance were similar to the changes in crude protein degradability and to the changes of solubility in SC and AM. The in situ method proved very suitable for the determination of the extent of degradability of the feed proteins in the rumen.

  12. Does Dietary Mitigation of Enteric Methane Production Affect Rumen Function and Animal Productivity in Dairy Cows?

    PubMed Central

    Veneman, Jolien B.; Muetzel, Stefan; Hart, Kenton J.; Faulkner, Catherine L.; Moorby, Jon M.; Perdok, Hink B.; Newbold, Charles J.

    2015-01-01

    It has been suggested that the rumen microbiome and rumen function might be disrupted if methane production in the rumen is decreased. Furthermore concerns have been voiced that geography and management might influence the underlying microbial population and hence the response of the rumen to mitigation strategies. Here we report the effect of the dietary additives: linseed oil and nitrate on methane emissions, rumen fermentation, and the rumen microbiome in two experiments from New Zealand (Dairy 1) and the UK (Dairy 2). Dairy 1 was a randomized block design with 18 multiparous lactating cows. Dairy 2 was a complete replicated 3 x 3 Latin Square using 6 rumen cannulated, lactating dairy cows. Treatments consisted of a control total mixed ration (TMR), supplementation with linseed oil (4% of feed DM) and supplementation with nitrate (2% of feed DM) in both experiments. Methane emissions were measured in open circuit respiration chambers and rumen samples were analyzed for rumen fermentation parameters and microbial population structure using qPCR and next generation sequencing (NGS). Supplementation with nitrate, but not linseed oil, decreased methane yield (g/kg DMI; P<0.02) and increased hydrogen (P<0.03) emissions in both experiments. Furthermore, the effect of nitrate on gaseous emissions was accompanied by an increased rumen acetate to propionate ratio and consistent changes in the rumen microbial populations including a decreased abundance of the main genus Prevotella and a decrease in archaeal mcrA (log10 copies/ g rumen DM content). These results demonstrate that methane emissions can be significantly decreased with nitrate supplementation with only minor, but consistent, effects on the rumen microbial population and its function, with no evidence that the response to dietary additives differed due to geography and different underlying microbial populations. PMID:26509835

  13. Does Dietary Mitigation of Enteric Methane Production Affect Rumen Function and Animal Productivity in Dairy Cows?

    PubMed

    Veneman, Jolien B; Muetzel, Stefan; Hart, Kenton J; Faulkner, Catherine L; Moorby, Jon M; Perdok, Hink B; Newbold, Charles J

    2015-01-01

    It has been suggested that the rumen microbiome and rumen function might be disrupted if methane production in the rumen is decreased. Furthermore concerns have been voiced that geography and management might influence the underlying microbial population and hence the response of the rumen to mitigation strategies. Here we report the effect of the dietary additives: linseed oil and nitrate on methane emissions, rumen fermentation, and the rumen microbiome in two experiments from New Zealand (Dairy 1) and the UK (Dairy 2). Dairy 1 was a randomized block design with 18 multiparous lactating cows. Dairy 2 was a complete replicated 3 x 3 Latin Square using 6 rumen cannulated, lactating dairy cows. Treatments consisted of a control total mixed ration (TMR), supplementation with linseed oil (4% of feed DM) and supplementation with nitrate (2% of feed DM) in both experiments. Methane emissions were measured in open circuit respiration chambers and rumen samples were analyzed for rumen fermentation parameters and microbial population structure using qPCR and next generation sequencing (NGS). Supplementation with nitrate, but not linseed oil, decreased methane yield (g/kg DMI; P<0.02) and increased hydrogen (P<0.03) emissions in both experiments. Furthermore, the effect of nitrate on gaseous emissions was accompanied by an increased rumen acetate to propionate ratio and consistent changes in the rumen microbial populations including a decreased abundance of the main genus Prevotella and a decrease in archaeal mcrA (log10 copies/g rumen DM content). These results demonstrate that methane emissions can be significantly decreased with nitrate supplementation with only minor, but consistent, effects on the rumen microbial population and its function, with no evidence that the response to dietary additives differed due to geography and different underlying microbial populations.

  14. Effect of protein level and urea in concentrate mixture on feed intake and rumen fermentation in swamp buffaloes fed rice straw-based diet.

    PubMed

    Kang, Sungchhang; Wanapat, Metha; Phesatcha, Kampanat; Norrapoke, Thitima

    2015-04-01

    Four rumen-fistulated Thai native swamp buffaloes were randomly assigned according to a 2 × 2 factorial arrangement in a 4 × 4 Latin square design to assess the effect of protein (CP) level and urea (U) source in concentrate diet on feed utilization and rumen ecology. The treatments were as follows: concentrate containing CP at 120 g/kg (soybean meal, SBM) (T1), 160 g/kg (SBM) (T2), 120 g/kg (U) (T3), and 160 g/kg (U) (T4), respectively. All buffaloes were fed concentrate at 10 g/kg of body weight, and rice straw was offered ad libitum. Feed intake and digestibilities of CP, neutral detergent fiber, and acid detergent fiber increased (P < 0.05) in treatments with higher level of CP especially with U source (P < 0.05). In contrast, CP level and source in concentrate did not affect on ruminal pH and temperature (P > 0.05), while concentration of ruminal ammonia (N), blood urea (U), volatile fatty acids profile, microorganism populations, and variable bacterial growth increased in buffaloes consumed concentrate containing CP at 160 g/kg (T2 and T4; P < 0.05). Fecal and urinary N excretions decreased in buffaloes consumed concentrate containing higher CP level especially with U source while purine derivatives increased which resulted in a higher N balance as compared to lower CP level and SBM source treatments (P < 0.05). In summary, higher CP level in concentrate improved feed intake, nutrient digestibility, purine derivatives, and rumen ecology, and U had shown better result than SBM. Concentrate mixtures containing 16 g/kg CP with U 40 g/kg could improved nutrients utilization with no adverse effects for swamp buffaloes fed on rice straw.

  15. Gene expression in bovine rumen epithelium during weaning identifies molecular regulators of rumen development and growth.

    PubMed

    Connor, Erin E; Baldwin, Ransom L; Li, Cong-jun; Li, Robert W; Chung, Hoyoung

    2013-03-01

    During weaning, epithelial cell function in the rumen transitions in response to conversion from a pre-ruminant to a true ruminant environment to ensure efficient nutrient absorption and metabolism. To identify gene networks affected by weaning in bovine rumen, Holstein bull calves were fed commercial milk replacer only (MRO) until 42 days of age, then were provided diets of either milk + orchardgrass hay (MH) or milk + grain-based calf starter (MG). Rumen epithelial RNA was extracted from calves sacrificed at four time points: day 14 (n = 3) and day 42 (n = 3) of age while fed the MRO diet and day 56 (n = 3/diet) and day 70 (n = 3/diet) while fed the MH and MG diets for transcript profiling by microarray hybridization. Five two-group comparisons were made using Permutation Analysis of Differential Expression® to identify differentially expressed genes over time and developmental stage between days 14 and 42 within the MRO diet, between day 42 on the MRO diet and day 56 on the MG or MH diets, and between the MG and MH diets at days 56 and 70. Ingenuity Pathway Analysis (IPA) of differentially expressed genes during weaning indicated the top 5 gene networks involving molecules participating in lipid metabolism, cell morphology and death, cellular growth and proliferation, molecular transport, and the cell cycle. Putative genes functioning in the establishment of the rumen microbial population and associated rumen epithelial inflammation during weaning were identified. Activation of transcription factor PPAR-α was identified by IPA software as an important regulator of molecular changes in rumen epithelium that function in papillary development and fatty acid oxidation during the transition from pre-rumination to rumination. Thus, molecular markers of rumen development and gene networks regulating differentiation and growth of rumen epithelium were identified for selecting targets and methods for improving and assessing rumen development and

  16. Effect of total mixed ration composition on fermentation and efficiency of ruminal microbial crude protein synthesis in vitro.

    PubMed

    Boguhn, J; Kluth, H; Rodehutscord, M

    2006-05-01

    The goal of this study was to identify dietary factors that affect fermentation and efficiency of microbial crude protein (CP(M)) synthesis in the rumen in vitro. We used 16 total mixed, dairy cow rations with known digestibilities that varied in ingredient composition and nutrient content. Each ration was incubated in a Rusitec (n = 3) for 15 d, and fermentation of different fractions was assessed. Observed extents of fermentation in 24 h were 35 to 47% for organic matter, 25 to 60% for crude protein, 3 to 28% for neutral detergent fiber, and 31 to 45% for gross energy. Organic matter fermentation depended on the content of crude protein and neutral detergent fiber in the ration. We studied net synthesis of CP(M) using an 15N dilution technique and found that 7 d of continuous 15N application are needed to achieve an 15N enrichment plateau in the N of isolated microbes in this type of study. The efficiency of CP(M) synthesis was 141 to 286 g/kg of fermented organic matter or 4.9 to 11.1 g/MJ of metabolizable energy, and these ranges agree with those found in the literature. Multiple regressions to predict the efficiency of CP(M) synthesis by diet data showed that crude protein was the only dietary chemical fraction that had a significant effect. Fat content and the inclusion rate of corn silage in the ration also tended to improve efficiency. We suggest that microbial need for preformed amino acids may explain the crude protein effect. A large part of the variation in efficiency of microbial activity still remains unexplained.

  17. Effects of rumen-undegradable protein sources and supplemental 2-hydroxy-4-(methylthio)-butanoic acid and lysine-HCl on lactation performance in dairy cows.

    PubMed

    Johnson-VanWieringen, L M; Harrison, J H; Davidson, D; Swift, M L; von Keyserlingk, M A G; Vazquez-Anon, M; Wright, D; Chalupa, W

    2007-11-01

    One hundred primiparous and multiparous Holstein cows were used in an experiment to evaluate the effect of supplementing diets with either a plant- or an animal-based source of rumen-undegradable protein (RUP), with or without AA supplementation, during the transition period and early lactation on milk production response. The experimental design was a randomized block design with approximately one-third of the cows being primiparous. Cows were assigned to 1 of 4 prepartum diets introduced 3 wk before the expected calving date and switched to the corresponding postpartum diet at calving. Diets 1 (AMI) and 2 (AMI+) included a vegetable RUP source (heat- and lignosulfonate-treated canola meal), with diet 2 containing supplemental Lys x HCl and Met hydroxy analog sources [D,L-2 hydroxy-4-(methylthio)-butanoic acid; Alimet feed supplement]. Diets 3 (PRO) and 4 (PRO+) consisted of a blend of animal RUP sources (blood meal, fish meal, feather meal, and porcine meat and bone meal), with diet 4 containing supplemental Lys x HCl and Met hydroxy analog sources [D,L-2 hydroxy-4-(methylthio)-butanoic acid; Alimet]. During the first 4 wk of lactation, dry matter intake was less when synthetic Lys x HCl and Alimet were supplemented, but this effect was no longer evident in wk 5 to 9 of the experiment. Interestingly, despite the initial decrease in dry matter intake in the cows fed AA-supplemented diets, there was no effect of treatment on milk production or the ratio of fat-corrected milk to dry matter intake throughout the 17 wk of the study. Undegradable protein source (vegetable vs. animal) did not affect dry matter intake, milk production, or 3.5% fat-corrected milk production for the first 17 wk of lactation. The results of this study indicate that heat- and lignosulfonate-treated canola meal can be used as a source of undegradable protein in place of high-quality rumen-undegradable animal protein sources without negative effects on milk production when diets are equivalent

  18. An Integrated Multi-Omics Approach Reveals the Effects of Supplementing Grass or Grass Hay with Vitamin E on the Rumen Microbiome and Its Function

    PubMed Central

    Belanche, Alejandro; Kingston-Smith, Alison H.; Newbold, Charles J.

    2016-01-01

    Rumen function is generally suboptimal leading to losses in methane and nitrogen. Analysis of the rumen microbiome is thus important to understanding the underlying microbial activity under different feeding strategies. This study investigated the effect of forage conservation method and vitamin E supplementation on rumen function using a rumen simulation technique. Ryegrass (GRA) or ryegrass hay (HAY) was supplemented with 20% concentrate containing zero or 50 IU/d vitamin E, as α-tocopheryl acetate, according to a 2 × 2 factorial design. The forage conservation method did not substantially change the nutrient composition but had a profound impact on the structure and diversity of the rumen microbiome. HAY diets promoted a more complex bacterial community (+38 OTUs) dominated by Firmicutes. This bacterial adaptation, together with increased rumen protozoa levels and methanogen diversity, was associated with greater fiber disappearance (+12%) in HAY diets, but also with greater rumen true N degradability (+7%) than GRA diets. HAY diets also had a higher metabolic H recovery and methane production (+35%) suggesting more efficient inter-species H transfer between bacteria, protozoa and methanogens. Contrarily, GRA diets promoted more simplified methanogen and bacterial communities, which were dominated by Bacteroidetes and Lactobacillus, thus lactate formation may have acted as an alternative H sink in GRA diets. Moreover the structure of the bacterial community with GRA diets was highly correlated with N utilization, and GRA diets promoted greater bacterial growth and microbial protein synthesis (+16%), as well as a more efficient microbial protein synthesis (+22%). A dose-response experiment using batch cultures revealed that vitamin E supplementation increased rumen fermentation in terms of total VFA and gas production, with protozoal activity higher when supplying α-tocopheryl acetate vs. α-tocopherol. Moreover, α-tocopheryl acetate promoted a small increase

  19. The giraffe (Giraffa camelopardalis) rumen microbiome.

    PubMed

    Roggenbuck, Michael; Sauer, Cathrine; Poulsen, Morten; Bertelsen, Mads F; Sørensen, Søren J

    2014-10-01

    Recent studies have shown that wild ruminants are sources of previously undescribed microorganisms, knowledge of which can improve our understanding of the complex microbial interactions in the foregut. Here, we investigated the microbial community of seven wild-caught giraffes (Giraffa camelopardalis), three of which were fed natural browse and four were fed Boskos pellets, leafy alfalfa hay, and cut savanna browse, by characterizing the 16S rRNA gene diversity using 454 FLX high-throughput sequencing. The microbial community composition varied according to diet, but differed little between the ruminal fluid and solid fraction. The giraffe rumen contained large levels of the phyla of Firmicutes and Bacteroidetes independent of diet, while Prevotella, Succinclasticium, and Methanobrevibacter accounted for the largest abundant taxonomic assigned genera. However, up to 21% of the generated sequences could not been assigned to any known bacterial phyla, and c. 70% not to genus, revealing that the giraffe rumen hosts a variety of previously undescribed bacteria.

  20. Linseed oil and DGAT1 K232A polymorphism: Effects on methane emission, energy and nitrogen metabolism, lactation performance, ruminal fermentation, and rumen microbial composition of Holstein-Friesian cows.

    PubMed

    van Gastelen, S; Visker, M H P W; Edwards, J E; Antunes-Fernandes, E C; Hettinga, K A; Alferink, S J J; Hendriks, W H; Bovenhuis, H; Smidt, H; Dijkstra, J

    2017-09-13

    Complex interactions between rumen microbiota, cow genetics, and diet composition may exist. Therefore, the effect of linseed oil, DGAT1 K232A polymorphism (DGAT1), and the interaction between linseed oil and DGAT1 on CH4 and H2 emission, energy and N metabolism, lactation performance, ruminal fermentation, and rumen bacterial and archaeal composition was investigated. Twenty-four lactating Holstein-Friesian cows (i.e., 12 with DGAT1 KK genotype and 12 with DGAT1 AA genotype) were fed 2 diets in a crossover design: a control diet and a linseed oil diet (LSO) with a difference of 22 g/kg of dry matter (DM) in fat content between the 2 diets. Both diets consisted of 40% corn silage, 30% grass silage, and 30% concentrates (DM basis). Apparent digestibility, lactation performance, N and energy balance, and CH4 emission were measured in climate respiration chambers, and rumen fluid samples were collected using the oral stomach tube technique. No linseed oil by DGAT1 interactions were observed for digestibility, milk production and composition, energy and N balance, CH4 and H2 emissions, and rumen volatile fatty acid concentrations. The DGAT1 KK genotype was associated with a lower proportion of polyunsaturated fatty acids in milk fat, and with a higher milk fat and protein content, and proportion of saturated fatty acids in milk fat compared with the DGAT1 AA genotype, whereas the fat- and protein-corrected milk yield was unaffected by DGAT1. Also, DGAT1 did not affect nutrient digestibility, CH4 or H2 emission, ruminal fermentation or ruminal archaeal and bacterial concentrations. Rumen bacterial and archaeal composition was also unaffected in terms of the whole community, whereas at the genus level the relative abundances of some bacterial genera were found to be affected by DGAT1. The DGAT1 KK genotype was associated with a lower metabolizability (i.e., ratio of metabolizable to gross energy intake), and with a tendency for a lower milk N efficiency compared with the

  1. Detection of Proteobacteria from the rumen by PCR using methanotroph-specific primers.

    PubMed

    Mitsumori, M; Ajisaka, N; Tajima, K; Kajikawa, H; Kurihara, M

    2002-01-01

    To detect Proteobacteria, including methanotrophs, from the rumen fluid and the bacteria inhabiting the rumen epithelium. Proteobacteria inhabiting the rumen were detected by PCR using methanotroph-specific primers. The detected Proteobacteria were divided into clusters A, B, and C in addition to one clone, which was distinct from the clusters and closely related to Nitrosomonas sp. The clusters A, B, and C were close to Succinivibrio dextrinosolvens, Enterobacter cloacae, and Actinobacillus minor, respectively. The clones obtained from the rumen fluid each belonged to cluster A or B. The clones obtained from the rumen epithelium belonged to cluster B or C or to Nitrosomonas sp. It has been assumed that the rumen fluid and the rumen epithelium host different populations of Proteobacteria. Moreover, detection of Nitrosomonas from the rumen epithelium would indicate the possibility that the bacterium oxidizes ammonia and methane on the rumen surface. These findings suggest that the rumen fluid and the epithelium support different microbial populations, which would play specific roles in rumen function. Future study should focus on the relationship between these communities and physiological functions in the rumen.

  2. Detect changes in protein structure of carinata meal during rumen fermentation in relation to basic chemical profile and comparison with canola meal using ATR-FT/IR molecular spectroscopy with chemometrics

    NASA Astrophysics Data System (ADS)

    Xin, Hangshu; Yu, Peiqiang

    2013-08-01

    As far as we know, no study has been carried out on whether protein structure changes in the feed during rumen fermentation from other research team. This study was conducted to characterize protein structure spectral changes in carinata meal during ruminal fermentation using Fourier transform infrared spectroscopy (FT/IR) technique with ATR. The objectives were to find out whether (1) protein internal structure (in terms of protein amide profile and protein secondary structure profile) changed after in situ ruminal fermentation at 0, 12, 24 and 48 h in carinata meal and conventional canola meal was used as a reference; (2) there was any correlation between protein spectral parameters and basic chemical profile in in situ rumen residue samples; and (3) the protein structural chemical make-up of carinata meal differed from canola meal during 48 h rumen incubation. The results showed that protein structure features in both carinata meal and canola meal were altered as incubation time increased (P < 0.0001) and linear and curvilinear relationships (P < 0.05) on amide II height and area, height and area ratio of amide I and II as well as height ratio of α-helix and β-sheet were observed within 48 h ruminal fermentation. And the amide I height and area as well as α-helix height and β-sheet height were in the highest level of IR absorbance at 0 h and then gradually declined linearly (P < 0.0001) by 30-38% after 48 h incubation. These results indicated that not only quantities decreased but also inherent structure changed in protein chemical make-up during ruminal fermentation. Meanwhile, strong correlations were found between protein spectral parameters and some basic nutrients profile such as CP (positively) and NDF (negatively). And both AHCA and PCA results showed that in situ rumen residues from carinata meal was not distinguished from those from canola meal, suggesting some relationship in structural make-up exhibited between them within protein region during 48

  3. Detect changes in protein structure of carinata meal during rumen fermentation in relation to basic chemical profile and comparison with canola meal using ATR-FT/IR molecular spectroscopy with chemometrics.

    PubMed

    Xin, Hangshu; Yu, Peiqiang

    2013-08-01

    As far as we know, no study has been carried out on whether protein structure changes in the feed during rumen fermentation from other research team. This study was conducted to characterize protein structure spectral changes in carinata meal during ruminal fermentation using Fourier transform infrared spectroscopy (FT/IR) technique with ATR. The objectives were to find out whether (1) protein internal structure (in terms of protein amide profile and protein secondary structure profile) changed after in situ ruminal fermentation at 0, 12, 24 and 48 h in carinata meal and conventional canola meal was used as a reference; (2) there was any correlation between protein spectral parameters and basic chemical profile in in situ rumen residue samples; and (3) the protein structural chemical make-up of carinata meal differed from canola meal during 48 h rumen incubation. The results showed that protein structure features in both carinata meal and canola meal were altered as incubation time increased (P<0.0001) and linear and curvilinear relationships (P<0.05) on amide II height and area, height and area ratio of amide I and II as well as height ratio of α-helix and β-sheet were observed within 48 h ruminal fermentation. And the amide I height and area as well as α-helix height and β-sheet height were in the highest level of IR absorbance at 0 h and then gradually declined linearly (P<0.0001) by 30-38% after 48 h incubation. These results indicated that not only quantities decreased but also inherent structure changed in protein chemical make-up during ruminal fermentation. Meanwhile, strong correlations were found between protein spectral parameters and some basic nutrients profile such as CP (positively) and NDF (negatively). And both AHCA and PCA results showed that in situ rumen residues from carinata meal was not distinguished from those from canola meal, suggesting some relationship in structural make-up exhibited between them within protein region during 48 h

  4. Metagenomics of rumen bacteriophage from thirteen lactating dairy cattle

    PubMed Central

    2013-01-01

    Background The bovine rumen hosts a diverse and complex community of Eukarya, Bacteria, Archea and viruses (including bacteriophage). The rumen viral population (the rumen virome) has received little attention compared to the rumen microbial population (the rumen microbiome). We used massively parallel sequencing of virus like particles to investigate the diversity of the rumen virome in thirteen lactating Australian Holstein dairy cattle all housed in the same location, 12 of which were sampled on the same day. Results Fourteen putative viral sequence fragments over 30 Kbp in length were assembled and annotated. Many of the putative genes in the assembled contigs showed no homology to previously annotated genes, highlighting the large amount of work still required to fully annotate the functions encoded in viral genomes. The abundance of the contig sequences varied widely between animals, even though the cattle were of the same age, stage of lactation and fed the same diets. Additionally the twelve animals which were co-habited shared a number of their dominant viral contigs. We compared the functional characteristics of our bovine viromes with that of other viromes, as well as rumen microbiomes. At the functional level, we found strong similarities between all of the viral samples, which were highly distinct from the rumen microbiome samples. Conclusions Our findings suggest a large amount of between animal variation in the bovine rumen virome and that co-habiting animals may have more similar viromes than non co-habited animals. We report the deepest sequencing to date of the rumen virome. This work highlights the enormous amount of novelty and variation present in the rumen virome. PMID:24180266

  5. Metabolism of Peptides by Rumen Microorganisms

    PubMed Central

    Wright, D. E.

    1967-01-01

    Rumen microorganisms utilize tryptic peptides from Chlorella protein, forming carbon dioxide, volatile fatty acids, and bacterial protein. Peptide carbon is more efficiently converted into bacterial protein than is amino acid carbon. A progressive degradation of the peptides was demonstrated by use of columns of Sephadex G-25. PMID:6035045

  6. Variability of Actinobacteria, a minor component of rumen microflora.

    PubMed

    Suľák, M; Sikorová, L; Jankuvová, J; Javorský, P; Pristaš, P

    2012-07-01

    Actinobacteria (Actinomycetes) are a significant and interesting group of gram-positive bacteria. They are regular, though infrequent, members of the microbial life in the rumen and represent up to 3 % of total rumen bacteria; there is considerable lack of information about ecology and biology of rumen actinobacteria. During the characterization of variability of rumen treponemas using non-cultivation approach, we also noted the variability of rumen actinobacteria. By using Treponema-specific primers a specific 16S rRNA gene library was prepared from cow and sheep rumen total DNA. About 10 % of recombinant clones contained actinobacteria-like sequences. Phylogenetic analyses of 11 clones obtained showed the high variability of actinobacteria in the ruminant digestive system. While some sequences are nearly identical to known sequences of actinobacteria, we detected completely new clusters of actinobacteria-like sequences, representing probably new, as yet undiscovered, group of rumen Actinobacteria. Further research will be necessary for understanding their nature and functions in the rumen.

  7. Fluorescent proteins in microbial biotechnology--new proteins and new applications.

    PubMed

    Vizcaino-Caston, Isaac; Wyre, Chris; Overton, Tim W

    2012-02-01

    The recent advances over the past 5 years in the utilisation of fluorescent proteins in microbial biotechnology applications, including recombinant protein production, food processing, and environmental biotechnology, are reviewed. We highlight possible areas where fluorescent proteins currently used in other bioscience disciplines could be adapted for use in biotechnological applications and also outline novel uses for recently developed fluorescent proteins.

  8. Role and function of short chain fatty acids in rumen epithelial metabolism, development and importance of the rumen epithelium in understanding control of transcriptome

    USDA-ARS?s Scientific Manuscript database

    The epithelial lining of the rumen is uniquely placed to have impact on the nutrient metabolism of the animal. The symbiotic relationship with the microbial populations that inhabit the rumen, serves to provide a constant supply of nutrients from roughage that would otherwise be unusable. Metaboli...

  9. Symbiosis and Rumen Protozoa

    ERIC Educational Resources Information Center

    Dillon, Raymond D.

    1970-01-01

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

  10. Symbiosis and Rumen Protozoa

    ERIC Educational Resources Information Center

    Dillon, Raymond D.

    1970-01-01

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

  11. Effect of supplemental concentrate during the dry period or early lactation on rumen epithelium gene and protein expression in dairy cattle during the transition period.

    PubMed

    Dieho, K; van Baal, J; Kruijt, L; Bannink, A; Schonewille, J T; Carreño, D; Hendriks, W H; Dijkstra, J

    2017-09-01

    We previously reported 2 experiments with rumen-cannulated Holstein-Friesian dairy cows showing that during the transition period, rumen papillae surface area, and fractional absorption rate of volatile fatty acids (VFA) increase after calving. However, supplemental concentrate during the dry period and rate of increase of concentrate allowance during lactation affected papillae surface area, but not VFA absorption. Here we report the changes in gene and protein expression in rumen papillae related to tissue growth and VFA utilization. The lactation experiment treatment consisted of a rapid [RAP; 1.0 kg of dry matter (DM)/d; n = 6] or gradual (GRAD; 0.25 kg of DM/d; n = 6) increase of concentrate allowance (up to 10.9 kg of DM/d), starting at 4 d postpartum (pp). The dry period experiment treatment consisted of 3.0 kg of DM/d of concentrate (n = 4) or no concentrate (n = 5) during the last 28 d of the dry period. Real-time quantitative PCR analysis of rumen papillae showed that the expression of apoptosis-related genes was neither affected by day nor its interaction with treatment for both experiments. Expression of epithelial transporter genes was not affected by day or treatment in the lactation experiment, except for NBC1. In the dry period experiment, expression of MCT1, NBC1, DRA, NHE2, NHE3, and UT-B generally decreased after calving. A day and treatment interaction was observed for ATP1A1 in the dry period experiment, with greater expression at 18 and 8 d antepartum for concentrate than no concentrate. Generally, expression of VFA metabolism-related genes was not affected by day or its interaction with treatment. In the lactation experiment, immunoblotting of 5 selected genes showed that protein expression of DRA and PCCA was greater at 16 d pp compared with 3 and 44 d pp. Expression of NHE2 was greater, and that of ATP1A1 lower, at 16 and 44 d pp compared with 3 d pp, suggesting alterations in intracellular pH regulation and sodium homeostasis. Both MCT1

  12. Microbial degradation of dissolved proteins in seawater

    SciTech Connect

    Hollibaugh, J.T.; Azam, F.

    1983-11-01

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

  13. Rumen-protected methionine and lysine: effects on milk production and plasma amino acids of dairy cows with reference to metabolisable protein status.

    PubMed

    Awawdeh, Mofleh S

    2016-05-01

    Two experiments were conducted to study the effects of rumen-protected Met (RPM) alone or with rumen-protected Lys (RPL) on milk yield and plasma amino acids of dairy cows. In experiment 1, 24 multiparous Holstein cows (154 DIM) were assigned to one of 3 groups where each cow received 0 g/d of RPM and RPL (C), 30 g/d of RPM (M), or 30 g/d of RPM plus 25 g of RPL (ML). The study lasted for 8 weeks where milk yield and composition were determined weekly. Daily milk yield averaged 28·0, 27·8, and 29·7 kg/cow for the C, M, and ML groups, respectively. Dietary treatments had no effects (P ≥ 0·54) on milk contents of fat, lactose, solid non-fat or total solids. Milk protein content in the ML group was greater (P < 0·05) than the C and M groups. Plasma levels of all AA were not significantly (P ≥ 0·09) affected by supplemental RPL and/or RPM. In experiment 2, 30 multiparous Holstein cows (100 DIM) were assigned to one of 3 groups where each cow received 0 g/d of RPM and RPL (C), 50 g/d of RPM (M), or 50 g/d of RPM plus 25 g/d of RPL (ML). The study lasted for 5 weeks. Cows in the M (30·5 kg) and ML (31·4 kg) groups produced (P < 0·05) more milk than those of the C group (29·1 kg). Under conditions of this study, RPM plus RPL improved milk yield and protein contents of dairy cows and was better than supplying RPM alone. Response in milk yield to RPM and RPL was affected by the MP status of cows which deserves further investigation.

  14. Microbial Protein-Antigenome Determination (MAD) Technology: A Proteomics-Based Strategy for Rapid Identification of Microbial Targets of Host Humoral Immune Responses

    USDA-ARS?s Scientific Manuscript database

    Immunogenic, pathogen-specific proteins have excellent potential for development of novel management modalities. Here, we describe an innovative application of proteomics called Microbial protein-Antigenome Determination (MAD) Technology for rapid identification of native microbial proteins that eli...

  15. Microbial Protein-Antigenome Determination (MAD) Technology: A Proteomics-Based Strategy for Rapid Identification of Microbial Targets of Host Humoral Immune Responses

    USDA-ARS?s Scientific Manuscript database

    Immunogenic, pathogen-specific proteins have excellent potential for development of novel management modalities. Here, we describe an innovative application of proteomics called Microbial protein-Antigenome Determination (MAD) Technology for rapid identification of native microbial proteins that el...

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

    PubMed

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

    2017-01-18

    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 (P<0.01) and microbial nitrogen synthesis in the rumen tended to decrease by adding CA to barley. Treating barley grain with citric 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).

  17. Effect of dietary protein level and rumen-protected amino acid supplementation on amino acid utilization for milk protein in lactating dairy cows.

    PubMed

    Lee, C; Giallongo, F; Hristov, A N; Lapierre, H; Cassidy, T W; Heyler, K S; Varga, G A; Parys, C

    2015-03-01

    This study investigated the effect of metabolizable protein (MP) supply and rumen-protected (RP) Lys and Met supplementation on productivity, nutrient digestibility, urinary N losses, apparent total-tract digestibility of dietary AA, and the efficiency of AA utilization for milk protein synthesis in dairy cows. The experiment was conducted with 8 ruminally cannulated Holstein cows in a replicated 4×4 Latin square design trial with 21-d periods. Treatments were (1) MP-adequate diet (AMP; MP balance of -24 g/d); (2) MP-deficient diet (DMP; MP balance of -281 g/d); (3) DMP supplemented with 100 g of RPLys/cow per day (estimated digestible Lys supply=24 g/d; DMPL; MP balance of -305g/d); and (4) DMPL supplemented with 24 g of RPMet/cow per day (estimated digestible Met supply=15 g/d; DMPLM; MP balance of -256g/d). Diet had no effect on total-tract nutrient digestibility, milk production, and milk composition, but the DMP diets decreased urinary N excretion and the ammonia emitting potential of manure. Plasma Met concentration was increased by DMPLM compared with AMP. Supplementation with RPLys had no effect on plasma Lys. Concentration of most AA in milk protein was increased or tended to be increased by DMPLM compared with DMPL. Except for the AA supplemented as RPAA (i.e., Met and Lys), apparent total-tract digestibility of all dietary AA was generally greater for the DMP diets and ranged from 33% (Arg, AMP diet) to 67% (Thr, DMPL diet). Apparent recovery of dietary AA in milk protein followed the same trends, being greater for the DMP diets than AMP and generally lower for Lys and Met with the RPAA-supplemented diets versus AMP and DMP. The RPAA were apparently not used for milk protein synthesis in the conditions of this experiment. The AA recoveries in milk protein varied from around 17% (Ala) to 70% (Pro). Milk protein recoveries of essential AA (EAA) were around 54% for the DMP diet and 49% for AMP. The estimated efficiency of utilization of digestible EAA for

  18. High throughput whole rumen metagenome profiling using untargeted massively parallel sequencing

    PubMed Central

    2012-01-01

    Background Variation of microorganism communities in the rumen of cattle (Bos taurus) is of great interest because of possible links to economically or environmentally important traits, such as feed conversion efficiency or methane emission levels. The resolution of studies investigating this variation may be improved by utilizing untargeted massively parallel sequencing (MPS), that is, sequencing without targeted amplification of genes. The objective of this study was to develop a method which used MPS to generate “rumen metagenome profiles”, and to investigate if these profiles were repeatable among samples taken from the same cow. Given faecal samples are much easier to obtain than rumen fluid samples; we also investigated whether rumen metagenome profiles were predictive of faecal metagenome profiles. Results Rather than focusing on individual organisms within the rumen, our method used MPS data to generate quantitative rumen micro-biome profiles, regardless of taxonomic classifications. The method requires a previously assembled reference metagenome. A number of such reference metagenomes were considered, including two rumen derived metagenomes, a human faecal microflora metagenome and a reference metagenome made up of publically available prokaryote sequences. Sequence reads from each test sample were aligned to these references. The “rumen metagenome profile” was generated from the number of the reads that aligned to each contig in the database. We used this method to test the hypothesis that rumen fluid microbial community profiles vary more between cows than within multiple samples from the same cow. Rumen fluid samples were taken from three cows, at three locations within the rumen. DNA from the samples was sequenced on the Illumina GAIIx. When the reads were aligned to a rumen metagenome reference, the rumen metagenome profiles were repeatable (P < 0.00001) by cow regardless of location of sampling rumen fluid. The repeatability was estimated

  19. Effects of Supplementation of Eucalyptus (E. Camaldulensis) Leaf Meal on Feed Intake and Rumen Fermentation Efficiency in Swamp Buffaloes

    PubMed Central

    Thao, N. T.; Wanapat, M.; Kang, S.; Cherdthong, A.

    2015-01-01

    Four rumen fistulated swamp buffaloes were randomly assigned according to a 4×4 Latin square design to investigate the effects of Eucalyptus (E. Camaldulensis) leaf meal (ELM) supplementation as a rumen enhancer on feed intake and rumen fermentation characteristics. The dietary treatments were as follows: T1 = 0 g ELM/hd/d; T2 = 40 g ELM/hd/d; T3 = 80 g ELM/hd/d; T4 = 120 g ELM/hd/d, respectively. Experimental animals were kept in individual pens and concentrate was offered at 0.3% BW while rice straw was fed ad libitum. The results revealed that voluntary feed intake and digestion coefficients of nutrients were similar among treatments. Ruminal pH, temperature and blood urea nitrogen concentrations were not affected by ELM supplementation; however, ELM supplementation resulted in lower concentration of ruminal ammonia nitrogen. Total volatile fatty acids, propionate concentration increased with the increasing level of EML (p<0.05) while the proportion of acetate was decreased (p<0.05). Methane production was linearly decreased (p<0.05) with the increasing level of ELM supplementation. Protozoa count and proteolytic bacteria population were reduced (p<0.05) while fungal zoospores and total viable bacteria, amylolytic, cellulolytic bacteria were unchanged. In addition, nitrogen utilization and microbial protein synthesis tended to increase by the dietary treatments. Based on the present findings, it is suggested that ELM could modify the rumen fermentation and is potentially used as a rumen enhancer in methane mitigation and rumen fermentation efficiency. PMID:26104399

  20. Insights into Abundant Rumen Ureolytic Bacterial Community Using Rumen Simulation System

    PubMed Central

    Jin, Di; Zhao, Shengguo; Wang, Pengpeng; Zheng, Nan; Bu, Dengpan; Beckers, Yves; Wang, Jiaqi

    2016-01-01

    Urea, a non-protein nitrogen for dairy cows, is rapidly hydrolyzed to ammonia by urease produced by ureolytic bacteria in the rumen, and the ammonia is used as nitrogen for rumen bacterial growth. However, there is limited knowledge with regard to the ureolytic bacteria community in the rumen. To explore the ruminal ureolytic bacterial community, urea, or acetohydroxamic acid (AHA, an inhibitor of urea hydrolysis) were supplemented into the rumen simulation systems. The bacterial 16S rRNA genes were sequenced by Miseq high-throughput sequencing and used to reveal the ureoltyic bacteria by comparing different treatments. The results revealed that urea supplementation significantly increased the ammonia concentration, and AHA addition inhibited urea hydrolysis. Urea supplementation significantly increased the richness of bacterial community and the proportion of ureC genes. The composition of bacterial community following urea or AHA supplementation showed no significant difference compared to the groups without supplementation. The abundance of Bacillus and unclassified Succinivibrionaceae increased significantly following urea supplementation. Pseudomonas, Haemophilus, Neisseria, Streptococcus, and Actinomyces exhibited a positive response to urea supplementation and a negative response to AHA addition. Results retrieved from the NCBI protein database and publications confirmed that the representative bacteria in these genera mentioned above had urease genes or urease activities. Therefore, the rumen ureolytic bacteria were abundant in the genera of Pseudomonas, Haemophilus, Neisseria, Streptococcus, Actinomyces, Bacillus, and unclassified Succinivibrionaceae. Insights into abundant rumen ureolytic bacteria provide the regulation targets to mitigate urea hydrolysis and increase efficiency of urea nitrogen utilization in ruminants. PMID:27446045

  1. Suppressive subtractive hybridization as a tool for identifying genetic diversity in an environmental metagenome: the rumen as a model.

    PubMed

    Galbraith, Elizabeth A; Antonopoulos, Dionysios A; White, Bryan A

    2004-09-01

    Molecular techniques previously used for genome comparisons of closely related bacterial species could prove extremely valuable for comparisons of complex microbial communities, or metagenomes. Our study aimed to determine the breadth and value of suppressive subtractive hybridization (SSH) in a pilot-scale analysis of metagenomic DNA from communities of microorganisms in the rumen. Suppressive subtractive hybridization was performed using total genomic DNA isolated from rumen fluid samples of two hay-fed steers, arbitrarily designated as tester or driver. Ninety-six subtraction DNA fragments from the tester metagenome were amplified, cloned and the DNA sequences were determined. Verification of the isolation of DNA fragments unique to the tester metagenome was accomplished through dot blot and Southern blot hybridizations. Tester-specific SSH fragments were found in 95 of 96 randomly selected clones. DNA sequences of subtraction fragments were analysed by computer assisted DNA and amino acid comparisons. Putative translations of 26 (32.1%) subtractive hybridization fragments exhibited significant similarity to Bacterial proteins, whereas 15 (18.5%) distinctive subtracted fragments had significant similarity to proteins from Archaea. The remainder of the subtractive hybridization fragments displayed no similarity to GenBank sequences. This metagenomic approach has exposed an unexpectedly large difference in Archaeal community structure between the rumen microbial populations of two steers fed identical diets and housed together. 16S rRNA dot blot hybridizations revealed similar proportions of Bacteria and Archaea in both rumen samples and suggest that the differences uncovered by SSH are the result of varying community structural composition. Our study demonstrates a novel approach to comparative analyses of environmental microbial communities through the use of SSH.

  2. Variation of in situ rumen degradation of crude protein and amino acids and in vitro digestibility of undegraded feed protein in rapeseed meals.

    PubMed

    Steingass, H; Kneer, G; Wischer, G; Rodehutscord, M

    2013-07-01

    In this study, 10 samples of rapeseed meal (RSM) from 10 different oil plants in Germany were examined. In situ rumen degradation of CP was determined by incubation over 1, 2, 4, 8, 16, 32 and 72 h in duplicate per time point using three rumen fistulated dry cows. Degradation kinetics were estimated by an exponential model and effective CP degradation was calculated. Degradation was corrected for small particle loss as the difference between washing loss and water-soluble fraction. Amino acid analysis was carried out in the samples and in the residues after 8 and 16 h of incubation in situ and degradation of individual amino acids was calculated for these incubation times. In vitro pepsin-pancreatin digestibility of CP (IPD) was determined in the samples as well as in the 8 and 16 h residues. Effective CP degradation for a rumen outflow rate of 8%/h (ED8) averaged 54.3% with a considerable variation among samples ranging from 44.3% to 62.7%. A multiple regression equation containing acid detergent insoluble N, total glucosinolates and petroleum ether extract as independent variables predicted ED8 with satisfying accuracy (R 2 = 0.74; RSD = 6.4%). Degradation of amino acids was different from that of CP for most amino acids studied, especially after 8 h of incubation. Compared with CP, degradation of essential amino acids was predominantly lower while degradation of non-essential amino acids was higher in most cases. However, for lysine and methionine no distinct difference with CP degradation was found. Degradation of individual amino acids was predicted from CP degradation with high accuracy using linear regression equations. Average IPD of RSM was 79.8 ± 2.6%. IPD was lower in the incubation residues and decreased with longer incubation time and increasing rumen degradation, respectively.

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

    PubMed

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

    2012-10-01

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

  4. Performance of beef cows and calves fed different sources of rumen-degradable protein when grazing stockpiled limpograss pastures.

    PubMed

    Aguiar, A D; Vendramini, J M B; Arthington, J D; Sollenberger, L E; DiLorenzo, N; Hersom, M J

    2015-04-01

    Two experiments evaluated the effects of different sources of RDP on forage characteristics, animal performance, and ruminal and blood parameters of beef cattle grazing stockpiled limpograss (Hemarthria altissima) from January to May 2011 and 2012. In Exp. 1, 24 mature lactating beef cows and their respective calves were allocated to 8 stockpiled limpograss pastures (3 pairs/pasture). Treatments were 2 different sources of RDP, urea or cottonseed (Gossypium spp.) meal (CSM), distributed in a completely randomized design with 4 replicates. Feather meal and corn (Zea mays) meal were added to the urea treatments to balance RUP and energy. Treatments were mixed in sugarcane (Saccharum officinarum) molasses, which resulted in 3 kg DM/cow per day of supplement. There were no differences (P > 0.10) in herbage mass (HM; 3,200 ± 400 kg DM/ha), herbage allowance (HA; 1.9 ± 0.2 kg DM/kg of BW), CP (5.2 ± 0.2%), and in vitro digestible organic matter (IVDOM; 47 ± 0.5%) concentrations. There was a decrease (P < 0.10) in HM (from 4,100 to 2,600 kg/ha), IVDOM (from 46 to 39.9%), and HA (from 2.5 to 1.4 kg DM/kg BW) from January to March. Cow ADG (0.23 ± 0.08 kg/d), BCS (4.6 ± 0.2), milk yield (7.0 ± 0.4 kg/d), and plasma urea nitrogen (PUN; 16.1 ± 0.8 mg/dL) and calf ADG (0.71 ± 0.05 kg/d) were similar (P > 0.10) among treatments. Sixteen cow-calf pairs were moved to 8 drylot pens after Exp. 1, maintained on the same treatment, and evaluated for forage and total DMI. There was no difference in forage (P = 0.16; 2.1 ± 0.1% BW) and total DMI (P = 0.12; 2.5 ± 0.1% BW) between treatments. In Exp. 2, 2 rumen-cannulated steers were used in a 2 × 2 Latin square design, replicated in 2011 and 2012, to test the effects of the same treatments on rumen fluid and blood parameters. There was no difference (P > 0.10) in ruminal NH3-N (12.9 ± 0.3 mg/dL), pH (6.5 ± 0.1), propionic acid (25 ± 2.2 mol/100 mol), acetic acid (69.2 ± 2.9 mol/100 mol), and butyric acid (4.5 ± 0.5 mol

  5. Rumen microbiota and dietary fat: a mutual shaping.

    PubMed

    Enjalbert, F; Combes, S; Zened, A; Meynadier, A

    2017-10-01

    Although fat content in usual ruminant diets is very low, fat supplements can be given to farm ruminants to modulate rumen activity or the fatty acid (FA) profile of meat and milk. Unsaturated FAs, which are dominant in common fat sources for ruminants, have negative effects on microbial growth, especially protozoa and fibrolytic bacteria. In turn, the rumen microbiota detoxifies unsaturated FAs (UFAs) through a biohydrogenation (BH) process, transforming dietary UFAs with cis geometrical double-bonds into mainly trans UFAs and, finally, into saturated FAs. Culture studies have provided a large amount of data regarding bacterial species and strains that are affected by UFAs or involved in lipolysis or BH, with a major focus on the Butyrivibrio genus. More recent data using molecular approaches to rumen microbiota extend and challenge these data, but further research will be necessary to improve our understanding of fat and rumen microbiota interactions. © 2017 The Society for Applied Microbiology.

  6. The complete genome sequence of Eubacterium limosum SA11, a metabolically versatile rumen acetogen.

    PubMed

    Kelly, William J; Henderson, Gemma; Pacheco, Diana M; Li, Dong; Reilly, Kerri; Naylor, Graham E; Janssen, Peter H; Attwood, Graeme T; Altermann, Eric; Leahy, Sinead C

    2016-01-01

    Acetogens are a specialized group of anaerobic bacteria able to produce acetate from CO2 and H2 via the Wood-Ljungdahl pathway. In some gut environments acetogens can compete with methanogens for H2, and as a result rumen acetogens are of interest in the development of microbial approaches for methane mitigation. The acetogen Eubacterium limosum SA11 was isolated from the rumen of a New Zealand sheep and its genome has been sequenced to examine its potential application in methane mitigation strategies, particularly in situations where hydrogenotrophic methanogens are inhibited resulting in increased H2 levels in the rumen. The 4.15 Mb chromosome of SA11 has an average G + C content of 47 %, and encodes 3805 protein-coding genes. There is a single prophage inserted in the chromosome, and several other gene clusters appear to have been acquired by horizontal transfer. These include genes for cell wall glycopolymers, a type VII secretion system, cell surface proteins and chemotaxis. SA11 is able to use a variety of organic substrates in addition to H2/CO2, with acetate and butyrate as the principal fermentation end-products, and genes involved in these metabolic pathways have been identified. An unusual feature is the presence of 39 genes encoding trimethylamine methyltransferase family proteins, more than any other bacterial genome. Overall, SA11 is a metabolically versatile organism, but its ability to grow on such a wide range of substrates suggests it may not be a suitable candidate to take the place of hydrogen-utilizing methanogens in the rumen.

  7. Effects of correcting in situ ruminal microbial colonization of feed particles on the relationship between ruminally undegraded and intestinally digested crude protein in concentrate feeds.

    PubMed

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

    2017-07-07

    In situ estimates of ruminally undegraded protein (RUP) and intestinally digested protein (IDP) of ten concentrates, uncorrected or corrected for the ruminal microbial colonization, were used to examine the effects of this correction on the relationship between IDP and RUP values. Both variables were established for three rumen and duodenum cannulated wethers using (15) N labeling-techniques and considering measured rates of ruminal particle comminution (kc ) and outflow (kp ). A covariance analysis showed that the close relationship found between both variables (IDP = -0.0132 ± 0.00679 + 0.776 ± 0.0002 RUP; n = 60; P < 0.001; r = 0.960) is not affected by correcting for microbial colonization (P = 0.682). The IDP content in concentrates and industrial by-products can be predicted from RUP values, thus avoiding the laborious and complex procedure of determining intestinal digestibility; however, a larger sample of feeds is necessary to achieve more accurate predictions. The lack of influence of the correction for microbial contamination on the prediction observed in the present study increases the data available for this prediction. However, only the use of corrected values may provide an accurate evaluation. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  8. Effects of different tannin-rich extracts and rapeseed tannin monomers on methane formation and microbial protein synthesis in vitro.

    PubMed

    Wischer, G; Boguhn, J; Steingaß, H; Schollenberger, M; Rodehutscord, M

    2013-11-01

    Tannins, polyphenolic compounds found in plants, are known to complex with proteins of feed and rumen bacteria. This group of substances has the potential to reduce methane production either with or without negative effects on digestibility and microbial yield. In the first step of this study, 10 tannin-rich extracts from chestnut, mimosa, myrabolan, quebracho, sumach, tara, valonea, oak, cocoa and grape seed, and four rapeseed tannin monomers (pelargonidin, catechin, cyanidin and sinapinic acid) were used in a series of in vitro trials using the Hohenheim gas test, with grass silage as substrate. The objective was to screen the potential of various tannin-rich extracts to reduce methane production without a significant effect on total gas production (GP). Supplementation with pelargonidin and cyanidin did not reduce methane production; however, catechin and sinapinic acid reduced methane production without altering GP. All tannin-rich extracts, except for tara extract, significantly reduced methane production by 8% to 28% without altering GP. On the basis of these results, five tannin-rich extracts were selected and further investigated in a second step using a Rusitec system. Each tannin-rich extract (1.5 g) was supplemented to grass silage (15 g). In this experiment, nutrient degradation, microbial protein synthesis and volatile fatty acid production were used as additional response criteria. Chestnut extract caused the greatest reduction in methane production followed by valonea, grape seed and sumach, whereas myrabolan extract did not reduce methane production. Whereas chestnut extract reduced acetate production by 19%, supplementation with grape seed or myrabolan extract increased acetate production. However, degradation of fibre fractions was reduced in all tannin treatments. Degradation of dry matter and organic matter was also reduced by tannin supplementation, and no differences were found between the tannin-rich extracts. CP degradation and ammonia

  9. Rumen metabolism of swamp buffaloes fed rice straw supplemented with cassava hay and urea.

    PubMed

    Ampapon, Thiwakorn; Wanapat, Metha; Kang, Sungchhang

    2016-04-01

    The objectives of this experiment were to investigate effects of cassava hay (CH) and urea (U) supplementation on feed intake, digestibility, rumen fermentation, and microbial protein synthesis of swamp buffaloes fed on rice straw. Four rumen-fistulated swamp buffaloes, 365 ± 15.0 kg, were randomly assigned according to a 4 × 4 Latin square design to receive four dietary treatments: T1 = CH 400 g/head/day + U 0 g/head/day, T2 = CH + U 30 g/head/day, T3 = CH + U 60 g/head/day, and T4 = CH + U 90 g/head/day, respectively. Results revealed that feed intake was not affected while nutrient digestibilities were increased (P < 0.05) with increasing U level supplementation especially at 90 g/head/day. Ruminal pH and temperature were not altered by urea supplementation, whereas ammonia nitrogen (NH3-N) and blood urea nitrogen were increased with urea supplement (P < 0.05). In addition, total volatile fatty acid and butyric acid were similar among treatments, while propionic acid (C3) was increased by level of urea supplement (P < 0.05), but acetic acid (C2) and C2/C3 ratio were significantly decreased (P < 0.05). On the other hand, protozoal population and methane production were decreased by CH and urea supplement, while bacterial population particularly those of proteolytic, cellulolytic, and amylolytic bacteria and efficiency of microbial nitrogen synthesis were linearly increased (P < 0.05). Based on this experiment, it suggested that supplementation of urea and cassava hay for buffaloes fed rice straw improved rumen ecology and increased fermentation end products and microbial protein synthesis while reducing protozoal populations and methane production. Urea supplements of 60-90 g/head/day when fed with cassava hay are recommended for swamp buffaloes consuming rice straw.

  10. Rumen microbial response in production of CLA and methane to safflower oil in association with fish oil or/and fumarate.

    PubMed

    Li, Xiang Z; Long, Rui J; Yan, Chang G; Lee, Hong G; Kim, Young J; Song, Man K

    2011-06-01

    Supplementation effect of fish oil and/or fumarate on production of conjugated linoleic acid (CLA) and methane by rumen microbes was examined when incubated with safflower oil. One hundred and twenty milligrams of safflower oil (SO), safflower oil with 24 mg fish oil (SOFO), safflower oil with 24 mmol/L fumarate (SOFA), or safflower oil with 24 mg fish oil and 24 mmol/L fumarate (SOFOFA) were added to the 90 mL culture solution. The culture solution was also made without any supplements (control). The SOFA and SOFOFA increased pH and propionate (C3) compared to other treatments from 3 h incubation time. An accumulated amount of total methane (CH(4) ) for 12 h incubation was decreased by all the supplements compared to control. The concentrations of c9,t11CLA for all the incubation times were increased in the treatments of SOFO, SOFA and SOFOFA compared to SO. The highest concentration of c9,t11CLA was observed from SOFOFA among all the treatments at all incubation times. Overall data indicate that supplementation of combined fumarate and/or fish oil when incubated with safflower oil could depress CH(4) generation and increase production of C(3) and CLA under the condition of current in vitro study.

  11. Manipulation of Rumen Microbial Fermentation by Polyphenol Rich Solvent Fractions from Papaya Leaf to Reduce Green-House Gas Methane and Biohydrogenation of C18 PUFA.

    PubMed

    Jafari, Saeid; Meng, Goh Yong; Rajion, Mohamed Ali; Jahromi, Mohammad Faseleh; Ebrahimi, Mahdi

    2016-06-08

    Different solvents (hexane, chloroform, ethyl acetate, butanol, and water) were used to identify the effect of papaya leaf (PL) fractions (PLFs) on ruminal biohydrogenation (BH) and ruminal methanogenesis in an in vitro study. PLFs at a concentration of 0 (control, CON) and 15 mg/250 mg dry matter (DM) were mixed with 30 mL of buffered rumen fluid and were incubated for 24 h. Methane (CH4) production (mL/250 mg DM) was the highest (P < 0.05) for CON (7.65) and lowest for the chloroform fraction (5.41) compared to those of other PLFs at 24 h of incubation. Acetate to propionate ratio was the lowest for PLFs compared to that of CON. Supplementation of the diet with PLFs significantly (P < 0.05) decreased the rate of BH of C18:1n-9 (oleic acid; OA), C18:2n-6 (linoleic acid; LA), and C18:3n-3 (α-linolenic acid; LNA) compared to that of CON after 24 h of incubation. Real time PCR indicated that total protozoa and total methanogen population in PLFs decreased (P < 0.05) compared to those of CON.

  12. Perturbation Dynamics of the Rumen Microbiota in Response to Exogenous Butyrate

    PubMed Central

    Li, Robert W.; Wu, Sitao; Baldwin, Ransom L.; Li, Weizhong; Li, Congjun

    2012-01-01

    The capacity of the rumen microbiota to produce volatile fatty acids (VFAs) has important implications in animal well-being and production. We investigated temporal changes of the rumen microbiota in response to butyrate infusion using pyrosequencing of the 16S rRNA gene. Twenty one phyla were identified in the rumen microbiota of dairy cows. The rumen microbiota harbored 54.5±6.1 genera (mean ± SD) and 127.3±4.4 operational taxonomic units (OTUs), respectively. However, the core microbiome comprised of 26 genera and 82 OTUs. Butyrate infusion altered molar percentages of 3 major VFAs. Butyrate perturbation had a profound impact on the rumen microbial composition. A 72 h-infusion led to a significant change in the numbers of sequence reads derived from 4 phyla, including 2 most abundant phyla, Bacteroidetes and Firmicutes. As many as 19 genera and 43 OTUs were significantly impacted by butyrate infusion. Elevated butyrate levels in the rumen seemingly had a stimulating effect on butyrate-producing bacteria populations. The resilience of the rumen microbial ecosystem was evident as the abundance of the microorganisms returned to their pre-disturbed status after infusion withdrawal. Our findings provide insight into perturbation dynamics of the rumen microbial ecosystem and should guide efforts in formulating optimal uses of probiotic bacteria treating human diseases. PMID:22253719

  13. [In vitro estimation using radioactive phosphorus of the phosphorus requirements of rumen microorganisms].

    PubMed

    Durand, M; Beaumatin, P; Dumay, C

    1983-01-01

    Microbial requirements for P were assumed to be a function of the amount of microbial protein synthesis (microbial growth) and of the quantity of organic matter (OM) fermented in the rumen. The relationships among P incorporation into microbial matter and protein synthesis, ammonia utilization, volatile fatty acid (VFA) production and organic matter fermented (OMF) were studied in short-term incubations (3 h) using 32P-labelled phosphate. The amount of P incorporated was calculated from extracellular phosphate pool specific activity and the radioactivity incorporated into the microbial sediment during incubation (table 1). The inocula came from sheep fed a protein-free purified diet. In order to vary the intensity of fermentation, carbohydrates with a wide range of degrees of enzymatic susceptibility were used as substrates and the medium was either provided or was deficient in S and trace elements (table 4). Nitrogen was supplied as ammonium salts. Linear regression analyses showed that P incorporation was positively correlated with the criteria of protein synthesis and OM fermentation (figs. 1, 2, 3, 4). However, there was significant phosphorus incorporation when the value for nitrogen incorporation was zero (equation A: (Pi (mg) = 0.162 NH3-N + 0.376; r = 0.9). This was assumed to result either from energetic uncoupling (fermentation without concomitant bacterial growth) or from the lysis of cold microbial cells only. Equation A would reflect total P incorporation and equation A' Pi (mg) = 0.162 NH3-N (mg), net P incorporation. It was assumed that in vitro microbial requirements for P were in the range of 30-70 mg of P/liter of medium for 3-hour incubation, depending on the intensity of fermentation. From a mean value of microbial N yield of 30 g/kg of DOMR (organic matter apparently digested in the rumen), it was calculated that the total and net P requirements in vivo were 6 and 4.9 g/kg of DOMR, respectively, corresponding to 3.9 and 3.2 g/kg of DOM

  14. Amino acid composition of rumen bacteria and protozoa in cattle.

    PubMed

    Sok, M; Ouellet, D R; Firkins, J L; Pellerin, D; Lapierre, H

    2017-07-01

    Because microbial crude protein (MCP) constitutes more than 50% of the protein digested in cattle, its AA composition is needed to adequately estimate AA supply. Our objective was to update the AA contributions of the rumen microbial AA flowing to the duodenum using only studies from cattle, differentiating between fluid-associated bacteria (FAB), particle-associated bacteria (PAB), and protozoa, based on published literature (53, 16, and 18 treatment means were used for each type of microorganism, respectively). In addition, Cys and Met reported concentrations were retained only when an adequate protection of the sulfur groups was performed before the acid hydrolysis. The total AA (or true protein) fraction represented 82.4% of CP in bacteria. For 10 AA, including 4 essential AA, the AA composition differed between protozoa and bacteria. The most noticeable differences were a 45% lower Lys concentration and 40% higher Ala concentration in bacteria than in protozoa. Differences between FAB and PAB were less pronounced than differences between bacteria and protozoa. Assuming 33% FAB, 50% PAB, and 17% of protozoa in MCP duodenal flow, the updated concentrations of AA would decrease supply estimates of Met, Thr, and Val originating from MCP and increase those of Lys and Phe by 5 to 10% compared with those calculated using the FAB composition reported previously. Therefore, inclusion of the contribution of PAB and protozoa to the duodenal MCP flow is needed to adequately estimate AA supply from microbial origin when a factorial method is used to estimate duodenal AA flow. Furthermore, acknowledging the fact that hydrolysis of 1 kg of true microbial protein yields 1.16 kg of free AA substantially increases the estimates of AA supply from MCP. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  15. In situ protein degradation of alfalfa and birdsfoot trefoil hays and silages as influenced by condensed tannin concentration

    USDA-ARS?s Scientific Manuscript database

    Dairy cattle often make poor use of protein when offered diets comprised of high proportions of alfalfa (Medicago sativa L.) hay or silage because non-protein N (NPN) formed during forage conservation and ruminal fermentation exceeds requirements for rumen microbial protein synthesis; however, conde...

  16. Lotus corniculatus condensed tannins decrease in vivo populations of proteolytic bacteria and affect nitrogen metabolism in the rumen of sheep.

    PubMed

    Min, B R; Attwood, G T; Reilly, K; Sun, W; Peters, J S; Barry, T N; McNabb, W C

    2002-10-01

    . The Lotus corniculatus diet reduced rumen nitrogen digestibility (P < 0.05) and ammonia pool size and increased the flow of undegraded feed nitrogen to the abomasum. The nitrogen intake, rumen non-ammonia nitrogen pool size, rumen microbial non-ammonia nitrogen pool size, and abomasal microbial non-ammonia nitrogen fluxes were similar both in sheep fed only Lotus corniculatus and in sheep fed Lotus corniculatus plus PEG, but nonmicrobial non-ammonia nitrogen flux to the abomasum was higher (P < 0.01) for the sheep fed only Lotus corniculatus. Although condensed tannins in Lotus corniculatus reduced the populations of some proteolytic bacteria, total ruminal microbial protein and microbial protein outflow to the abomasum were unchanged, suggesting a species-specific effect of condensed tannins on bacteria in the rumen.

  17. Effects of feeding formate-treated alfalfa silage or red clover silage on omasal nutrient flow and microbial protein synthesis in lactating dairy cows.

    PubMed

    Brito, A F; Broderick, G A; Olmos Colmenero, J J; Reynal, S M

    2007-03-01

    Eight ruminally cannulated Holstein cows that were part of a larger lactation trial were blocked by days in milk and randomly assigned to replicated 4 x 4 Latin squares to quantify effects of nonprotein N (NPN) content of alfalfa silage (AS) and red clover silage (RCS) on omasal nutrient flows. Diets, fed as total mixed rations, contained 50% dry matter from control AS (CAS), ammonium tetraformate-treated AS (TAS), late maturity RCS (RCS1), or early maturity RCS (RCS2). Silages differed in NPN and acid detergent insoluble N (% of total N): 50 and 4% (CAS); 45 and 3% (TAS); 27 and 8% (RCS1); 29 and 4% (RCS2). The CAS, TAS, and RCS2 diets had 36% high-moisture shelled corn and 3% soybean meal, and the RCS1 diet had 31% high-moisture shelled corn and 9% soybean meal. All diets contained 10% corn silage, 27% neutral detergent fiber, and 17 to 18% crude protein. Compared with RCS, feeding AS increased the supply of rumen-degraded protein and omasal flows of nonammonia N and microbial protein, which may explain the improved milk yield observed in the companion lactation trial. However, omasal flow of rumen-undegraded protein was 34% greater on RCS. Except for Arg, omasal flows of individual AA, branched-chain AA, nonessential AA, essential AA, and total AA did not differ between cows fed AS vs. RCS. Within AS diets, no differences in omasal AA flows were observed. However, omasal flows of Asp, Ser, Glu, Cys, Val, Ile, Tyr, Lys, total nonessential AA, and total AA all were higher in cows fed RCS1 vs. cows fed RCS2. In this trial, there was no advantage to reducing NPN content of hay-crop silage.

  18. Effect of dietary nitrate level on enteric methane production, hydrogen emission, rumen fermentation, and nutrient digestibility in dairy cows.

    PubMed

    Olijhoek, D W; Hellwing, A L F; Brask, M; Weisbjerg, M R; Højberg, O; Larsen, M K; Dijkstra, J; Erlandsen, E J; Lund, P

    2016-08-01

    Nitrate may lower methane production in ruminants by competing with methanogenesis for available hydrogen in the rumen. This study evaluated the effect of 4 levels of dietary nitrate addition on enteric methane production, hydrogen emission, feed intake, rumen fermentation, nutrient digestibility, microbial protein synthesis, and blood methemoglobin. In a 4×4 Latin square design 4 lactating Danish Holstein dairy cows fitted with rumen, duodenal, and ileal cannulas were assigned to 4 calcium ammonium nitrate addition levels: control, low, medium, and high [0, 5.3, 13.6, and 21.1g of nitrate/kg of dry matter (DM), respectively]. Diets were made isonitrogenous by replacing urea. Cows were fed ad libitum and, after a 6-d period of gradual introduction of nitrate, adapted to the corn-silage-based total mixed ration (forage:concentrate ratio 50:50 on DM basis) for 16d before sampling. Digesta content from duodenum, ileum, and feces, and rumen liquid were collected, after which methane production and hydrogen emissions were measured in respiration chambers. Methane production [L/kg of dry matter intake (DMI)] linearly decreased with increasing nitrate concentrations compared with the control, corresponding to a reduction of 6, 13, and 23% for the low, medium, and high diets, respectively. Methane production was lowered with apparent efficiencies (measured methane reduction relative to potential methane reduction) of 82.3, 71.9, and 79.4% for the low, medium, and high diets, respectively. Addition of nitrate increased hydrogen emissions (L/kg of DMI) quadratically by a factor of 2.5, 3.4, and 3.0 (as L/kg of DMI) for the low, medium, and high diets, respectively, compared with the control. Blood methemoglobin levels and nitrate concentrations in milk and urine increased with increasing nitrate intake, but did not constitute a threat for animal health and human food safety. Microbial crude protein synthesis and efficiency were unaffected. Total volatile fatty acid

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

    PubMed

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

    2009-08-01

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

  20. Rumen microbial and fermentation characteristics are affected differently by acarbose addition during two nutritional types of simulated severe subacute ruminal acidosis in vitro.

    PubMed

    Wang, Yue; Liu, Junhua; Yin, Yuyang; Zhu, Weiyun; Mao, Shengyong

    2017-10-01

    Little information is available on whether or not the effect of an alpha-glucosidase inhibitor on the prevention of ruminal acidosis is influenced by the type of diet during ruminant feeding. This study was conducted to explore the effect of acarbose addition on the prevention of severe subacute ruminal acidosis induced by either cracked wheat or beet pulp in vitro. Cracked wheat and beet pulp were fermented in vitro by rumen microorganisms obtained from three dairy cows. When cracked wheat was used as the substrate and fermented for 24 h, compared with the control, acarbose addition decreased the concentrations of acetate, propionate, butyrate, total volatile fatty acids, and lactate (P < 0.05), while linearly increased the ratio of acetate to propionate, pH value, and the ammonia-nitrogen level (P < 0.05). Applying Illumina MiSeq sequencing of a fragment of the 16S rRNA gene revealed that the relative abundance of Firmicutes and Bacteroidetes as well as the ACE (abundance-based coverage estimator) value, Chao 1 value, and Shannon index increased significantly (P < 0.05), while there was a significant reduction (P < 0.05) in the relative abundance of Tenericutes as well as Proteobacteria after adding acarbose compared to the control. On the other hand, when beet pulp was used as the substrate, acarbose addition had no significant effects (P > 0.05) on the fermentation parameters and the Chao 1 value, the Shannon index, and the proportion of Firmicutes and Bacteroidetes. In general, these findings indicate that acarbose had more effects on ruminal fermentation when wheat was used as the substrate, whereas it exhibited little effect on ruminal fermentation when beet pulp was used as the substrate. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Short communication: Effects of increasing protein and energy in the milk replacer with or without direct-fed microbial supplementation on growth and performance of preweaned Holstein calves.

    PubMed

    Geiger, A J; Ward, S H; Williams, C C; Rude, B J; Cabrera, C J; Kalestch, K N; Voelz, B E

    2014-11-01

    Forty-four Holstein calves were fed a direct-fed microbial (DFM) and 1 of 2 milk replacers to evaluate calf performance and growth. Treatments were (1) a control milk replacer [22:20; 22% crude protein (CP) and 20% fat], (2) an accelerated milk replacer (27:10; 27% CP and 10% fat), (3) the control milk replacer with added DFM (22:20+D), and (4) the accelerated milk replacer with added DFM (27:10+D). Dry matter intake, rectal temperatures, respiration scores and rates, and fecal scores were collected daily. Body weight, hip and withers height, heart girth, blood, and rumen fluid samples were collected weekly. Effects of treatment, sex, week, and their interactions were analyzed. Calves fed an accelerated milk replacer, regardless of DFM supplementation, consumed more CP and metabolizable energy in the milk replacer. No treatment differences were found for starter intake or intake of neutral detergent fiber or acid detergent fiber in the starter. Calves fed the accelerated milk replacer had greater preweaning and weaning body weight compared with calves fed the control milk replacer. Average daily gain was greater during the preweaning period for calves fed the accelerated milk replacer, but the same pattern did not hold true during the postweaning period. Feed efficiency did not differ among treatments. Hip height tended to be and withers height and heart girth were greater at weaning for calves fed the accelerated milk replacer compared with calves fed the control milk replacer. Fecal scores were greatest in calves fed DFM. Overall acetate, propionate, butyrate, and n-valerate concentrations were lower in calves fed the accelerated milk replacer, but DFM did not have an effect. Rumen pH was not different. Blood metabolites were unaffected by DFM supplementation, but calves fed the accelerated milk replacer had increased partial pressure of CO2, bicarbonate, and total bicarbonate in the blood. Direct-fed microbial supplementation did not appear to benefit the calf

  2. Bovine somatotropin and rumen-undegradable protein effects in prepubertal dairy heifers: effects on body composition and organ and tissue weights.

    PubMed

    Moallem, U; Dahl, G E; Duffey, E K; Capuco, A V; Wood, D L; McLeod, K R; Baldwin, R L; Erdman, R A

    2004-11-01

    The objectives of this study were to determine the effect of recombinant bovine somatotropin (bST) and added dietary rumen undegradable protein (RUP) on organ and tissue weights and body composition in growing dairy heifers. Thirty-two Holstein heifers were in the experiment, 8 killed initially at 3 mo of age, with the remaining 24 Holstein heifers randomly assigned to treatments (n = 6) consisting of 0.1 mg/kg of body weight per day of bST and 2% added dietary RUP (dry matter basis) applied in a 2 x2 factorial design. A total of 6 heifers per treatment group (3 each at 5 and 10 mo of age), were slaughtered to determine body composition and organ masses. Feed intake measured from group intakes were increased by 0.25 and 0.35 kg/d with bST and RUP, respectively. Administration of bST tended to increase the weights of visceral organs including heart, kidney, and spleen by 16, 16, and 38%, respectively. At 10 mo of age, there was a trend for increased empty body weights (EBW) and non-carcass components for heifers treated with bST, but there were no effects of RUP. Body components and organ weights, expressed as a percentage of BW were not affected by RUP or bST. Somatotropin increased ash weight at 10 mo without affecting amounts of protein, fat, and energy. Rates of ash deposition between 3 and 10 mo of age were increased 7 and 4 g/d by bST and RUP, respectively. There were no treatment effects on rates of body fat, protein, and energy deposition. Bovine somatotropin and RUP altered the metabolism of growing heifers in a manner that was consistent with increased rates of skeletal growth. This suggests that nutritional and endocrine manipulations could increase growth rates of skeletal tissues without increasing fat deposition in prepubertal dairy heifers.

  3. Rumen management during aphagia.

    PubMed

    Shakespeare, A S

    2008-09-01

    Ruminants that for any reason are unable to eat enough to survive can be supported via rumen fistulation. To successfully accomplish this task, an understanding of rumen physiology is necessary. Some adaptation and modification of the normal physiological processes will be necessary because the extended time normally required to ingest food will, for obvious practical reasons, be reduced to a few minutes repeated once to three times a day. The physiology of significance to aphagic or dysphagic animals is discussed and relevant examples of clinical cases are used to illustrate practical applications.

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

    PubMed

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

    2017-10-01

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

  5. Rumen Microbiome from Steers Differing in Feed Efficiency

    PubMed Central

    2015-01-01

    The cattle rumen has a diverse microbial ecosystem that is essential for the host to digest plant material. Extremes in body weight (BW) gain in mice and humans have been associated with different intestinal microbial populations. The objective of this study was to characterize the microbiome of the cattle rumen among steers differing in feed efficiency. Two contemporary groups of steers (n=148 and n=197) were fed a ration (dry matter basis) of 57.35% dry-rolled corn, 30% wet distillers grain with solubles, 8% alfalfa hay, 4.25% supplement, and 0.4% urea for 63 days. Individual feed intake (FI) and BW gain were determined. Within contemporary group, the four steers within each Cartesian quadrant were sampled (n=16/group) from the bivariate distribution of average daily BW gain and average daily FI. Bacterial 16S rRNA gene amplicons were sequenced from the harvested bovine rumen fluid samples using next-generation sequencing technology. No significant changes in diversity or richness were indicated, and UniFrac principal coordinate analysis did not show any separation of microbial communities within the rumen. However, the abundances of relative microbial populations and operational taxonomic units did reveal significant differences with reference to feed efficiency groups. Bacteroidetes and Firmicutes were the dominant phyla in all ruminal groups, with significant population shifts in relevant ruminal taxa, including phyla Firmicutes and Lentisphaerae, as well as genera Succiniclasticum, Lactobacillus, Ruminococcus, and Prevotella. This study suggests the involvement of the rumen microbiome as a component influencing the efficiency of weight gain at the 16S level, which can be utilized to better understand variations in microbial ecology as well as host factors that will improve feed efficiency. PMID:26030887

  6. Control of rumen methanogenesis.

    PubMed

    Van Nevel, C J; Demeyer, D I

    1996-09-01

    During the last decades, considerable research on methane production in the rumen and its inhibition has been carried out. Initially, as methane production represents a significant loss of gross energy in the feed (2-15%), the ultimate goal of such intervention in rumen fermentation was an increase in feed efficiency. A second reason favouring research on methane inhibition is its role in the global warming phenomenon and in the destruction of the ozone layer. In this review, the authors describe briefly several interventions for reducing methane emission by ruminants. The objective can be reached by intervention at the dietary level by ration manipulation (composition, feeding level) or by the use of additives or supplements. Examples of additives are polyhalogenated compounds, ionophores and other antibiotics. Supplementation of the ration with lipids also lowered methanogenesis. More biotechnological interventions, e.g., defaunation, probiotics and introduction of reductive acetogenesis in the rumen, are also mentioned. It can be concluded that drastic inhibition of methane production is not unequivocally successful as a result of several factors, such as: instantaneous inhibition often followed by restoration of methanogenesis due to adaptation of the microbes or degradation of the additive, toxicity for the host animal, negative effects on overall digestion and productive performance. Therefore, methanogenesis and its inhibition cannot be considered as a separate part of rumen fermentation and its consequences on the animal should be taken into account.

  7. The bacterial community composition of the bovine rumen detected using pyrosequencing of 16S rRNA genes

    USDA-ARS?s Scientific Manuscript database

    The rumen as a complex microbial ecosystem plays a critical role in sustainable agriculture. Rumen microorganisms perform important biochemical conversions, including the fermentation of plant fiber to small molecules such as short-chain fatty acids for meat and dairy production. In this study, we s...

  8. Hydrogen production from proteins via electrohydrogenesis in microbial electrolysis cells.

    PubMed

    Lu, Lu; Xing, Defeng; Xie, Tianhui; Ren, Nanqi; Logan, Bruce E

    2010-08-15

    Microorganisms can produce hydrogen gas (H(2)) at high rates by fermentation of carbohydrates, but not from proteins. However, it is possible to produce H(2) at high rates and yields from proteins by electrohydrogenesis in microbial electrolysis cells (MECs). Hydrogen gas was generated using bovine serum albumin (BSA, 700 mg/L) in a single-chamber MEC at a rate of Q=0.42+/-0.07 m(3)/m(3)/day and a yield of Y(H2) = 21.0 +/- 5.0 mmol-H2/g-COD, with an energy recovery (relative to electrical input) of eta(E)=75+/-12% (applied voltage of 0.6 V). Hydrogen production was substantially reduced using a complex protein (peptone) under the same conditions, to Q=0.05+/-0.01 m(3)/m(3)/day, YH2 = 2.6 +/- 0.1 mmol-H2/g-COD, and eta(E)=14+/-3%. There was good removal of organic matter for both substrates in terms of either protein (87+/-6 -97 +/-2%) or total COD (86+/-2 - 91+/-2%). Electron recycling likely occurred as Coulombic efficiencies exceeded 100% using BSA. The use of a two-chamber design, with either a CEM or AEM membrane, reduced the hydrogen production rate, but did not appreciably affect the hydrogen yield or energy efficiency. When an MEC was first acclimated to acetate, and then switched to BSA, performance was substantially reduced and was similar to that obtained using peptone. These results demonstrate that electrohydrogenesis can be used to produce H(2) from proteins, and it can also be used as a method for treatment of protein-containing wastewaters.

  9. Effect of camelina oil or live yeasts (Saccharomyces cerevisiae) on ruminal methane production, rumen fermentation, and milk fatty acid composition in lactating cows fed grass silage diets.

    PubMed

    Bayat, A R; Kairenius, P; Stefański, T; Leskinen, H; Comtet-Marre, S; Forano, E; Chaucheyras-Durand, F; Shingfield, K J

    2015-05-01

    The potential of dietary supplements of 2 live yeast strains (Saccharomyces cerevisiae) or camelina oil to lower ruminal methane (CH4) and carbon dioxide (CO2) production and the associated effects on animal performance, rumen fermentation, rumen microbial populations, nutrient metabolism, and milk fatty acid (FA) composition of cows fed grass silage-based diets were examined. Four Finnish Ayrshire cows (53±7 d in milk) fitted with rumen cannula were used in a 4×4 Latin square with four 42-d periods. Cows received a basal total mixed ration (control treatment) with a 50:50 forage-to-concentrate ratio [on a dry matter (DM) basis] containing grass silage, the same basal total mixed ration supplemented with 1 of 2 live yeasts, A or B, administered directly in the rumen at 10(10) cfu/d (treatments A and B), or supplements of 60g of camelina oil/kg of diet DM that replaced concentrate ingredients in the basal total mixed ration (treatment CO). Relative to the control, treatments A and B had no effects on DM intake, rumen fermentation, ruminal gas production, or apparent total-tract nutrient digestibility. In contrast, treatment CO lowered DM intake and ruminal CH4 and CO2 production, responses associated with numerical nonsignificant decreases in total-tract organic matter digestibility, but no alterations in rumen fermentation characteristics or changes in the total numbers of rumen bacteria, methanogens, protozoa, and fungi. Compared with the control, treatment CO decreased the yields of milk, milk fat, lactose, and protein. Relative to treatment B, treatment CO improved nitrogen utilization due to a lower crude protein intake. Treatment A had no influence on milk FA composition, whereas treatment B increased cis-9 10:1 and decreased 11-cyclohexyl 11:0 and 24:0 concentrations. Treatment CO decreased milk fat 8:0 to 16:0 and total saturated FA, and increased 18:0, 18:1, 18:2, conjugated linoleic acid, 18:3n-3, and trans FA concentrations. Decreases in ruminal CH4

  10. Relationship between proteolysis in the silo and efficiency of utilization of dietary protein by lactating dairy cows

    USDA-ARS?s Scientific Manuscript database

    Ensiling is used widely to conserve forages for feeding to dairy cows. However, the protein in hay-crop silages is particularly susceptible to microbial breakdown in the rumen, and utilization of protein in alfalfa and grass silages by dairy cows is particularly poor. Dependent on maturity, hay-crop...

  11. MannDB: A microbial annotation database for protein characterization

    SciTech Connect

    Zhou, C; Lam, M; Smith, J; Zemla, A; Dyer, M; Kuczmarski, T; Vitalis, E; Slezak, T

    2006-05-19

    MannDB was created to meet a need for rapid, comprehensive automated protein sequence analyses to support selection of proteins suitable as targets for driving the development of reagents for pathogen or protein toxin detection. Because a large number of open-source tools were needed, it was necessary to produce a software system to scale the computations for whole-proteome analysis. Thus, we built a fully automated system for executing software tools and for storage, integration, and display of automated protein sequence analysis and annotation data. MannDB is a relational database that organizes data resulting from fully automated, high-throughput protein-sequence analyses using open-source tools. Types of analyses provided include predictions of cleavage, chemical properties, classification, features, functional assignment, post-translational modifications, motifs, antigenicity, and secondary structure. Proteomes (lists of hypothetical and known proteins) are downloaded and parsed from Genbank and then inserted into MannDB, and annotations from SwissProt are downloaded when identifiers are found in the Genbank entry or when identical sequences are identified. Currently 36 open-source tools are run against MannDB protein sequences either on local systems or by means of batch submission to external servers. In addition, BLAST against protein entries in MvirDB, our database of microbial virulence factors, is performed. A web client browser enables viewing of computational results and downloaded annotations, and a query tool enables structured and free-text search capabilities. When available, links to external databases, including MvirDB, are provided. MannDB contains whole-proteome analyses for at least one representative organism from each category of biological threat organism listed by APHIS, CDC, HHS, NIAID, USDA, USFDA, and WHO. MannDB comprises a large number of genomes and comprehensive protein sequence analyses representing organisms listed as high

  12. Chemical markers for rumen methanogens and methanogenesis.

    PubMed

    McCartney, C A; Bull, I D; Dewhurst, R J

    2013-06-01

    appears to be subject to considerable between-animal variation. This variation was also evident in the weak relationship between archaeol concentrations in rumen digesta and faeces. We speculate that variation in the distribution and kinetics of methanogens in the rumen may affect the survival and functioning of Archaea in the rumen and therefore contribute to genetic variation in methane production. Indeed, variation in the relationship between the numbers of micro-organisms present in the rumen and those leaving the rumen may explain variation in relationships between methane production and both milk fatty acid profiles and faecal archaeol. As a result, microbial markers in the faeces and milk are unlikely to relate well back to methanogenesis in the rumen. This work has also highlighted the need to describe methanogen abundance in all rumen fractions and this may explain the difficulty interpreting results on the basis of samples taken using stomach tubes or rumenocentesis.

  13. Selenite Reduction by Anaerobic Microbial Aggregates: Microbial Community Structure, and Proteins Associated to the Produced Selenium Spheres

    PubMed Central

    Gonzalez-Gil, Graciela; Lens, Piet N. L.; Saikaly, Pascal E.

    2016-01-01

    Certain types of anaerobic granular sludge, which consists of microbial aggregates, can reduce selenium oxyanions. To envisage strategies for removing those oxyanions from wastewater and recovering the produced elemental selenium (Se0), insights into the microbial community structure and synthesis of Se0 within these microbial aggregates are required. High-throughput sequencing showed that Veillonellaceae (c.a. 20%) and Pseudomonadaceae (c.a.10%) were the most abundant microbial phylotypes in selenite reducing microbial aggregates. The majority of the Pseudomonadaceae sequences were affiliated to the genus Pseudomonas. A distinct outer layer (∼200 μm) of selenium deposits indicated that bioreduction occurred in the outer zone of the microbial aggregates. In that outer layer, SEM analysis showed abundant intracellular and extracellular Se0 (nano)spheres, with some cells having high numbers of intracellular Se0 spheres. Electron tomography showed that microbial cells can harbor a single large intracellular sphere that stretches the cell body. The Se0 spheres produced by the microorganisms were capped with organic material. X-ray photoelectron spectroscopy (XPS) analysis of extracted Se0 spheres, combined with a mathematical approach to analyzing XPS spectra from biological origin, indicated that proteins and lipids were components of the capping material associated to the Se0 spheres. The most abundant proteins associated to the spheres were identified by proteomic analysis. Most of the proteins or peptide sequences capping the Se0 spheres were identified as periplasmic outer membrane porins and as the cytoplasmic elongation factor Tu protein, suggesting an intracellular formation of the Se0 spheres. In view of these and previous findings, a schematic model for the synthesis of Se0 spheres by the microorganisms inhabiting the granular sludge is proposed. PMID:27199909

  14. Selenite Reduction by Anaerobic Microbial Aggregates: Microbial Community Structure, and Proteins Associated to the Produced Selenium Spheres.

    PubMed

    Gonzalez-Gil, Graciela; Lens, Piet N L; Saikaly, Pascal E

    2016-01-01

    Certain types of anaerobic granular sludge, which consists of microbial aggregates, can reduce selenium oxyanions. To envisage strategies for removing those oxyanions from wastewater and recovering the produced elemental selenium (Se(0)), insights into the microbial community structure and synthesis of Se(0) within these microbial aggregates are required. High-throughput sequencing showed that Veillonellaceae (c.a. 20%) and Pseudomonadaceae (c.a.10%) were the most abundant microbial phylotypes in selenite reducing microbial aggregates. The majority of the Pseudomonadaceae sequences were affiliated to the genus Pseudomonas. A distinct outer layer (∼200 μm) of selenium deposits indicated that bioreduction occurred in the outer zone of the microbial aggregates. In that outer layer, SEM analysis showed abundant intracellular and extracellular Se(0) (nano)spheres, with some cells having high numbers of intracellular Se(0) spheres. Electron tomography showed that microbial cells can harbor a single large intracellular sphere that stretches the cell body. The Se(0) spheres produced by the microorganisms were capped with organic material. X-ray photoelectron spectroscopy (XPS) analysis of extracted Se(0) spheres, combined with a mathematical approach to analyzing XPS spectra from biological origin, indicated that proteins and lipids were components of the capping material associated to the Se(0) spheres. The most abundant proteins associated to the spheres were identified by proteomic analysis. Most of the proteins or peptide sequences capping the Se(0) spheres were identified as periplasmic outer membrane porins and as the cytoplasmic elongation factor Tu protein, suggesting an intracellular formation of the Se(0) spheres. In view of these and previous findings, a schematic model for the synthesis of Se(0) spheres by the microorganisms inhabiting the granular sludge is proposed.

  15. A Structural and Functional Elucidation of the Rumen Microbiome Influenced by Various Diets and Microenvironments

    PubMed Central

    Deusch, Simon; Camarinha-Silva, Amélia; Conrad, Jürgen; Beifuss, Uwe; Rodehutscord, Markus; Seifert, Jana

    2017-01-01

    abundant in the corn silage whereas oligosaccharide transporters showed a higher abundance in the fiber-rich diets. Proteins involved in carbon metabolism were detected in high numbers and identification of metabolites like short-chain fatty acids, methylamines and phenylpropionate by NMR enabled linkage between producers and products. This study forms a solid basis to retrieve deeper insight into the complex network of microbial adaptation in the rumen. PMID:28883813

  16. Comparative analysis of Escherichia coli O157 growth and protein-expression, in vitro and in vivo, in rumen fluid of cattle

    USDA-ARS?s Scientific Manuscript database

    Cattle are the primary reservoirs for Escherichia coli O157 (O157), a Shiga toxin-producing E. coli, with potential for serious extraintestinal sequelae in humans. In a recent study (Kudva IT et al. BMC Microbiol. 2014; 14:48), we reported that when cultured in rumen fluid from dairy cattle on the ...

  17. Fermentation of Ammonia Fiber Expansion Treated and Untreated Barley Straw in a Rumen Simulation Technique Using Rumen Inoculum from Cattle with Slow versus Fast Rate of Fiber Disappearance

    PubMed Central

    Griffith, Candace L.; Ribeiro, Gabriel O.; Oba, Masahito; McAllister, Tim A.; Beauchemin, Karen A.

    2016-01-01

    The purpose of this study was to determine the effect of rumen inoculum from heifers with fast vs. slow rate of in situ fiber digestion on the fermentation of complex versus easily digested fiber sources in the forms of untreated and Ammonia Fiber Expansion (AFEX) treated barley straw, respectively, using an artificial rumen simulation technique (Rusitec). In situ fiber digestion was measured in a previous study by incubating untreated barley straw in the rumen of 16 heifers fed a diet consisting of 700 g/kg barley straw and 300 g/kg concentrate. The two heifers with fastest rate of digestion (Fast ≥ 4.18% h-1) and the two heifers with the slowest rate of digestion (Slow ≤ 3.17% h-1) were chosen as inoculum donors for this study. Two Rusitec apparatuses each equipped with eight fermenters were used in a completely randomized block design with two blocks (apparatus) and four treatments in a 2 × 2 factorial arrangement of treatments (Fast or Slow rumen inoculum and untreated or AFEX treated straw). Fast rumen inoculum and AFEX straw both increased (P < 0.05) disappearance of dry matter (DMD), organic matter, true DMD, neutral detergent fiber, acid detergent fiber, and nitrogen (N) with an interactive effect between the two (P < 0.05). Fast rumen inoculum increased (P > 0.05) methane production per gram of digested material for both untreated and AFEX straw, and reduced (interaction, P < 0.05) acetate: propionate ratio for untreated straw. Greater relative populations of Ruminococcus albus (P < 0.05) and increased microbial N production (P = 0.045) were observed in Fast rumen inoculum. AFEX straw in Fast inoculum had greater total bacterial populations than Slow, but for untreated straw this result was reversed (interaction, P = 0.013). These findings indicate that differences in microbial populations in rumen fluid contribute to differences in the capacity of rumen inoculum to digest fiber. PMID:27899919

  18. Fermentation of Ammonia Fiber Expansion Treated and Untreated Barley Straw in a Rumen Simulation Technique Using Rumen Inoculum from Cattle with Slow versus Fast Rate of Fiber Disappearance.

    PubMed

    Griffith, Candace L; Ribeiro, Gabriel O; Oba, Masahito; McAllister, Tim A; Beauchemin, Karen A

    2016-01-01

    The purpose of this study was to determine the effect of rumen inoculum from heifers with fast vs. slow rate of in situ fiber digestion on the fermentation of complex versus easily digested fiber sources in the forms of untreated and Ammonia Fiber Expansion (AFEX) treated barley straw, respectively, using an artificial rumen simulation technique (Rusitec). In situ fiber digestion was measured in a previous study by incubating untreated barley straw in the rumen of 16 heifers fed a diet consisting of 700 g/kg barley straw and 300 g/kg concentrate. The two heifers with fastest rate of digestion (Fast ≥ 4.18% h(-1)) and the two heifers with the slowest rate of digestion (Slow ≤ 3.17% h(-1)) were chosen as inoculum donors for this study. Two Rusitec apparatuses each equipped with eight fermenters were used in a completely randomized block design with two blocks (apparatus) and four treatments in a 2 × 2 factorial arrangement of treatments (Fast or Slow rumen inoculum and untreated or AFEX treated straw). Fast rumen inoculum and AFEX straw both increased (P < 0.05) disappearance of dry matter (DMD), organic matter, true DMD, neutral detergent fiber, acid detergent fiber, and nitrogen (N) with an interactive effect between the two (P < 0.05). Fast rumen inoculum increased (P > 0.05) methane production per gram of digested material for both untreated and AFEX straw, and reduced (interaction, P < 0.05) acetate: propionate ratio for untreated straw. Greater relative populations of Ruminococcus albus (P < 0.05) and increased microbial N production (P = 0.045) were observed in Fast rumen inoculum. AFEX straw in Fast inoculum had greater total bacterial populations than Slow, but for untreated straw this result was reversed (interaction, P = 0.013). These findings indicate that differences in microbial populations in rumen fluid contribute to differences in the capacity of rumen inoculum to digest fiber.

  19. Protein folding and conformational stress in microbial cells producing recombinant proteins: a host comparative overview

    PubMed Central

    Gasser, Brigitte; Saloheimo, Markku; Rinas, Ursula; Dragosits, Martin; Rodríguez-Carmona, Escarlata; Baumann, Kristin; Giuliani, Maria; Parrilli, Ermenegilda; Branduardi, Paola; Lang, Christine; Porro, Danilo; Ferrer, Pau; Tutino, Maria Luisa; Mattanovich, Diethard; Villaverde, Antonio

    2008-01-01

    Different species of microorganisms including yeasts, filamentous fungi and bacteria have been used in the past 25 years for the controlled production of foreign proteins of scientific, pharmacological or industrial interest. A major obstacle for protein production processes and a limit to overall success has been the abundance of misfolded polypeptides, which fail to reach their native conformation. The presence of misfolded or folding-reluctant protein species causes considerable stress in host cells. The characterization of such adverse conditions and the elicited cell responses have permitted to better understand the physiology and molecular biology of conformational stress. Therefore, microbial cell factories for recombinant protein production are depicted here as a source of knowledge that has considerably helped to picture the extremely rich landscape of in vivo protein folding, and the main cellular players of this complex process are described for the most important cell factories used for biotechnological purposes. PMID:18394160

  20. The effects of frequency of feeding on some quantitative aspects of digestion in the rumens of growing steers.

    PubMed

    McAllan, A B; Lewis, P E; Griffith, E S

    1987-09-01

    Three steers with simple rumen and abomasal cannulas were given ground and pelleted diets containing predominantly dried grass meal (DG) or rolled barley (RB). Diets were given at frequencies of two or eight feeds/d in a simple changeover design. Chromic oxide and polyethylene glycol were given as flow markers and flows (g/24 h) of organic matter (OM), nitrogenous and carbohydrate compounds were calculated. Ribonucleic acid and 35S were used as microbial markers and diaminopimelic acid (DAP) as a bacterial marker. Frequency of feeding had no significant effect on mean rumen pH, ammonia levels or liquid outflow rates with either diet. Rumen volume was decreased and abomasal digesta flow increased on Diet DG with more feeds but these parameters were unaffected with Diet RB. Increased feeding frequency with both feeds resulted in increased numbers of protozoa. There were no significant effects of feeding frequency of Diet DG on the abomasal flows of any of the nitrogenous constituents measured. However, there was a significant increase in microbial-N flow from 33 to 43 g/d with more frequent feeding of diet RB which was not reflected in bacterial-N flow as measured by DAP. The apparent digestion of OM in the rumen, expressed as g/g intake with diet DG was 0.41 and 0.31 for two feeds and eight feeds/d respectively. Corresponding values for diet RB were 0.56 and 0.63 respectively. The reduction in OM digestion with frequent feeding of diet DG was reflected in similarly reduced rumen digestibilities of all dietary carbohydrate components whereas the increase in OM digestion with diet RB was reflected only by the component sugars of the dietary fibre. The efficiencies of microbial protein synthesis (expressed as gMN/kg ADOM) increased from 36 to 46 when the feeding frequency of diet DG was increased from two to eight times/d. No significant effect of frequency of feeding was found for diet RB. Mouth to abomasum degradation of feed-N (expressed as g/g intake) of 0.64 was

  1. Isolation and characterization of a non-specific endoglucanase from a metagenomic library of goat rumen.

    PubMed

    Cheng, Jianbo; Huang, Shuai; Jiang, Haiqin; Zhang, Yunhai; Li, Lvmu; Wang, Juhua; Fan, Caiyun

    2016-01-01

    A cellulase gene (cel28a) was isolated from a rumen microbial metagenome library of goat rumen microorganisms, cloned into E. coli, and expressed in active form. The gene has a length of 1596 bp obtained using a genome walking Kit and encodes a protein of 509 amino acids with a calculated MW of 55 kDa. The deduced amino acid sequence was homologous with cellulases belonging to the glycosyl hydrolase family 5 (GH5). The expressed protein showed activity toward carboxymethylcellulose (CMC) and xylan, suggesting non-specific endoglucanase activity. The optimal conditions for endoglucanase and xylanase activities were 50 °C and pH 5.0. The metal ions (Ca(2+), Fe(2+), Mn(2+) and Co(2+)) stimulated the cellulase activity of cel28a, while the other metal ions and chemicals (Ni(2+), Mg(2+), Zn(2+), Cu(2+), SDS and EDTA) inhibited the cellulase activity. Further examination of substrate preference showed a higher activity with CMC, oat spelt xylan and birchwood xylan than with filter paper and microcrystalline cellulose, again suggesting that the protein was an endoglucanase with xylanase activity.

  2. Microbial protein in soil: influence of extraction method and C amendment on extraction and recovery.

    PubMed

    Taylor, Erin B; Williams, Mark A

    2010-02-01

    The capacity to study the content and resolve the dynamics of the proteome of diverse microbial communities would help to revolutionize the way microbiologists study the function and activity of microorganisms in soil. To better understand the limitations of a proteomic approach to studying soil microbial communities, we characterized extractable soil microbial proteins using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Two methods were utilized to extract proteins from microorganisms residing in a Quitman and Benfield soil: (1) direct extraction of bulk protein from soil and (2) separation of the microorganisms from soil using density gradient centrifugation and subsequent extraction (DGC-EXT) of microbial protein. In addition, glucose and toluene amendments to soil were used to stimulate the growth of a subset of the microbial community. A bacterial culture and bovine serum albumin (BSA) were added to the soil to qualitatively assess their recovery following extraction. Direct extraction and resolution of microbial proteins using SDS-PAGE generally resulted in smeared and unresolved banding patterns on gels. DGC-EXT of microbial protein from soil followed by separation using SDS-PAGE, however, did resolve six to 10 bands in the Benfield but not the Quitman soil. DGC-EXT of microbial protein, but not direct extraction following the addition of glucose and toluene, markedly increased the number of bands (approximately 40) on the gels in both Benfield and Quitman soils. Low recoveries of added culture and BSA proteins using the direct extraction method suggest that proteins either bind to soil organic matter and mineral particles or that partial degradation takes place during extraction. Interestingly, DGC may have been preferentially selected for actively growing cells, as gauged by the 10-100x lower cy19:0/18:1omega7 ratio of the fatty acid methyl esters in the isolated community compared to that for the whole soil. DGC can be used to

  3. Effect of quebracho-chestnut tannin extracts at 2 dietary crude protein levels on performance, rumen fermentation, and nitrogen partitioning in dairy cows.

    PubMed

    Aguerre, M J; Capozzolo, M C; Lencioni, P; Cabral, C; Wattiaux, M A

    2016-06-01

    Our objective was to determine the effects of a tannin mixture extract on lactating cow performance, rumen fermentation, and N partitioning, and whether responses were affected by dietary crude protein (CP). The experiment was conducted as a split-plot with 24 Holstein cows (mean ± standard deviation; 669±55kg of body weight; 87±36 d in milk; 8 ruminally cannulated) randomly assigned to a diet of [dry matter (DM) basis] 15.3 or 16.6% CP (whole plot) and 0, 0.45, 0.90, or 1.80% of a tannin mixture in three 4×4 Latin squares within each level of CP (sub-plot). Tannin extract mixture was from quebracho and chestnut trees (2:1 ratio). Dietary CP level did not influence responses to tannin supplementation. A linear decrease in DM intake (25.5 to 23.4kg/d) was found, as well as a linear increase in milk/DM intake (1.62 to 1.75) and a trend for a linear decrease in fat-and-protein-corrected milk (38.4 to 37.1kg/d) with increasing levels of tannin supplementation. In addition, there was a negative linear effect for milk urea N (14.0 to 12.9mg/dL), milk protein yield (1.20 to 1.15kg), and concentration (2.87 to 2.83%). Furthermore, the change in milk protein concentration tended to be quadratic, and predicted maximum was 2.89% for a tannin mixture fed at 0.47% of dietary DM. Tannin supplementation reduced ruminal NH3-N (11.3 to 8.8mg/dL), total branched-chain volatile fatty acid concentration (2.97 to 2.47mol/100mol), DM, organic matter, CP, and neutral detergent fiber digestibility. Dietary tannin had no effect on intake N (587±63g/d), milk N (175±32g/d), or N utilization efficiency (29.7±4.4%). However, feeding tannin extracts linearly increased fecal N excretion (214 to 256g/d), but reduced urinary N (213 to 177g/d) and urinary urea N (141 to 116g/d) excretion. Decreasing dietary CP did not influence milk production, but increased N utilization efficiency (milk N/N intake; 0.27 to 0.33), and decreased milk urea N (15.4 to 11.8mg/dL), ruminal NH3-N (11.0 to 9.3mg

  4. The fate of glycerol entering the rumen of dairy cows and sheep.

    PubMed

    Werner Omazic, A; Kronqvist, C; Zhongyan, L; Martens, H; Holtenius, K

    2015-04-01

    This study investigated the fate of glycerol entering the rumen, in particular whether glycerol could be absorbed across the rumen epithelium. Three non-lactating rumen-fistulated cows were used to calculate the overall disappearance rate of glycerol in vivo and evaluate the rate of ruminal glycerol absorption. Rumen epithelial tissues isolated from sheep were used to characterise glycerol transport properties. The rate of rumen microbial degradation of glycerol was then studied in an in vitro system under anaerobic and thermo-regulated conditions. The results showed that glycerol can be absorbed from the rumen in significant amounts. The fractional rate of absorption of glycerol was not affected by variations in glycerol concentration in the buffer solution in the in vivo study. The glycerol absorption apparently occurred largely by passive diffusion and was probably not facilitated by carriers. Glycerol also disappeared via microbial digestion and outflow from the rumen through the omasal orifice. Journal of Animal Physiology and Animal Nutrition © 2014 Blackwell Verlag GmbH.

  5. Metaproteomics reveals functional shifts in microbial and human proteins during a preterm infant gut colonization case.

    PubMed

    Young, Jacque C; Pan, Chongle; Adams, Rachel M; Brooks, Brandon; Banfield, Jillian F; Morowitz, Michael J; Hettich, Robert L

    2015-10-01

    Microbial colonization of the human gastrointestinal tract plays an important role in establishing health and homeostasis. However, the time-dependent functional signatures of microbial and human proteins during early colonization of the gut have yet to be determined. To this end, we employed shotgun proteomics to simultaneously monitor microbial and human proteins in fecal samples from a preterm infant during the first month of life. Microbial community complexity increased over time, with compositional changes that were consistent with previous metagenomic and rRNA gene data. More specifically, the function of the microbial community initially involved biomass growth, protein production, and lipid metabolism, and then switched to more complex metabolic functions, such as carbohydrate metabolism, once the community stabilized and matured. Human proteins detected included those responsible for epithelial barrier function and antimicrobial activity. Some neutrophil-derived proteins increased in abundance early in the study period, suggesting activation of the innate immune system. Likewise, abundances of cytoskeletal and mucin proteins increased later in the time course, suggestive of subsequent adjustment to the increased microbial load. This study provides the first snapshot of coordinated human and microbial protein expression in a preterm infant's gut during early development.

  6. Metaproteomics Reveals Functional Shifts in Microbial and Human Proteins During a Preterm Infant Gut Colonization Case

    PubMed Central

    Young, Jacque C.; Pan, Chongle; Adams, Rachel; Brooks, Brandon; Banfield, Jillian F.; Morowitz, Michael J.; Hettich, Robert L.

    2015-01-01

    Microbial colonization of the human gastrointestinal tract plays an important role in establishing health and homeostasis. However, the time-dependent functional signatures of microbial and human proteins during early colonization of the gut have yet to be determined. To this end, we employed shotgun proteomics to simultaneously monitor microbial and human proteins in fecal samples from a preterm infant during the first month of life. Microbial community complexity increased over time, with compositional changes that were consistent with previous metagenomic and rRNA gene data. More specifically, the function of the microbial community initially involved biomass growth, protein production, and lipid metabolism, and then switched to more complex metabolic functions, such as carbohydrate metabolism, once the community stabilized and matured. Human proteins detected included those responsible for epithelial barrier function and antimicrobial activity. Some neutrophil-derived proteins increased in abundance early in the study period, suggesting activation of the innate immune system. Likewise, abundances of cytoskeletal and mucin proteins increased later in the time course, suggestive of subsequent adjustment to the increased microbial load. This study provides the first snapshot of coordinated human and microbial protein expression in a preterm infant’s gut during early development. PMID:26077811

  7. Comparison of feed intake, digestion and rumen function among domestic ruminant species grazing in upland vegetation communities.

    PubMed

    Ferreira, L M M; Hervás, G; Belenguer, A; Celaya, R; Rodrigues, M A M; García, U; Frutos, P; Osoro, K

    2016-04-14

    This study aimed to compare feed intake, digestion, rumen fermentation parameters and bacterial community of 5 beef cows, 12 crossed ewes and 12 goats grazing together in spring-early summer on heather-gorse vegetation communities with an adjacent area of improved pasture. Organic matter intake (OMI) and digestibility (OMD) were estimated using alkane markers. Ruminal fluid samples were collected for measuring fermentation parameters, and studying the bacterial community using terminal restriction fragment length polymorphism (T-RFLP). Spot samples of urine were taken to determine purine derivative (PD) and creatinine concentrations to estimate microbial protein synthesis in the rumen. Herbaceous species were the main dietary component in all animal species. Cattle had higher (p < 0.05) daily OMI (g/kg LW(0.75) ) and OMD, whereas sheep and goats showed similar values. The highest ammonia concentration was observed in sheep. Total VFA, acetate and butyrate concentrations were not influenced by animal species, while propionate concentrations in goats were 1.8 times lower (p < 0.05) than in sheep. Acetate:propionate ratio was greater (p < 0.05) in goats, whereas cattle excreted more allantoin (p < 0.05). Estimated supply of microbial N was higher in cows (p < 0.01), whereas the efficiency of microbial protein synthesis was lower (p < 0.01) in this animal species. Hierarchical clustering analysis indicated a clear effect of animal species on rumen bacterial structure. Differences among animal species were also observed in the relative frequency of several T-RFs. Certain T-RFs compatible with Lachnospiraceae, Proteobacteria and Clostridiales species were not found in goats, while these animals showed high relative frequencies of some fragments compatible with the Ruminococcaceae family that were not detected in sheep and cattle. Results suggest a close relationship between animals' grazing behaviour and rumen bacterial structure and its function. Goats seem

  8. Ruminal fermentation characteristics and microbial nitrogen assimilation in sheep fed differently composed grass silages.

    PubMed

    Seip, Katharina; Breves, Gerhard; Isselstein, Johannes; Daş, Gürbüz; Abel, Hansjörg

    2012-06-01

    The investigation aimed at examining if the composition of grassland silage affects the microbial nitrogen assimilation in the rumen of sheep. The silages were made of vegetative summer re-growths consisting of 48% grasses, 28% legumes and 24% other forbs (GCF) or of pure grass (G). Silage GCF contained more intermediately degradable non-structural and less slowly degradable carbohydrates, more crude protein (CP), a narrower ratio between slow and very slow degradable nitrogen (N), and exhibited higher in situ degradability of organic matter and CP than Silage G. Four adult wethers equipped with rumen fistulae were used in a two factorial trial. Feed was offered either as silage alone or as a mixture of silage and barley (60:40). Microbial N was estimated using continuous intraruminal 15N infusion and measurement of 15N-enrichment in microbes isolated from rumen liquor samples. With the exception of trends for ruminal butyrate concentrations, no interactions were detected between silage and barley feeding. Sheep receiving Silage GCF exhibited larger diurnal fluctuations of ammonia, and produced more microbial N (p < 0.05) than sheep on Silage G. Feeding the silages with barley decreased ruminal pH and elevated the concentrations of butyrate (p < 0.05). The 15N incorporation into microbial N was reduced by barley feeding (p < 0.05) along with a trend to accelerated rumen fluid turnover, resulting in similar microbial N yields as found in sheep receiving silage without barley. It is concluded that the larger and better balanced amounts of intermediately degradable carbohydrate- and N-containing fractions favoured the ruminal microbial protein synthesis in sheep consuming Silage GCF instead of Silage G.

  9. Metaproteomics Reveals Functional Shifts in Microbial and Human Proteins During Infant Gut Colonization Case

    SciTech Connect

    Young, Jacque C.; Pan, Chongle; Adams, Rachel M.; Brooks, Brandon; Banfield, Jillian F.; Morowitz, Michael J.; Robert L. Hettich

    2015-01-01

    The microbial colonization of the human gastrointestinal tract plays an important role in establishing health and homeostasis. However, the time-dependent functional signatures of microbial and human proteins during early colonization of the gut have yet to be determined. Thus, we employed shotgun proteomics to simultaneously monitor microbial and human proteins in fecal samples from a preterm infant during the first month of life. Microbial community complexity and functions increased over time, with compositional changes that were consistent with previous metagenomic and rRNA gene data indicating three distinct colonization phases. Overall microbial community functions were established relatively early in development and remained stable. Human proteins detected included those responsible for epithelial barrier function and antimicrobial activity. Some neutrophil-derived proteins increased in abundance early in the study period, suggesting activation of the innate immune system. Moreover, abundances of cytoskeletal and mucin proteins increased later in the time course, suggestive of subsequent adjustment to the increased microbial load. Our study provides the first snapshot of coordinated human and microbial protein expression in the infant gut during early development.

  10. Metaproteomics Reveals Functional Shifts in Microbial and Human Proteins During Infant Gut Colonization Case

    DOE PAGES

    Young, Jacque C.; Pan, Chongle; Adams, Rachel M.; ...

    2015-01-01

    The microbial colonization of the human gastrointestinal tract plays an important role in establishing health and homeostasis. However, the time-dependent functional signatures of microbial and human proteins during early colonization of the gut have yet to be determined. Thus, we employed shotgun proteomics to simultaneously monitor microbial and human proteins in fecal samples from a preterm infant during the first month of life. Microbial community complexity and functions increased over time, with compositional changes that were consistent with previous metagenomic and rRNA gene data indicating three distinct colonization phases. Overall microbial community functions were established relatively early in development andmore » remained stable. Human proteins detected included those responsible for epithelial barrier function and antimicrobial activity. Some neutrophil-derived proteins increased in abundance early in the study period, suggesting activation of the innate immune system. Moreover, abundances of cytoskeletal and mucin proteins increased later in the time course, suggestive of subsequent adjustment to the increased microbial load. Our study provides the first snapshot of coordinated human and microbial protein expression in the infant gut during early development.« less

  11. Research note: investigation on the metabolism of glycerol in the rumen of bulls.

    PubMed

    Kijora, C; Bergner, H; Götz, K P; Bartelt, J; Szakács, J; Sommer, A

    1998-01-01

    Two bulls, each fitted with rumen and duodenal cannulas, received (in addition to a hay-grain diet) twice daily an infusion of 200 g glycerol into the rumen over a period of six days. During this preliminary in vivo investigation, the influence of a glycerol application on the rumen environment over a six-day adaptation period was examined. Samples of rumen fluid were collected daily, two hours after glycerol infusion. An additional 15N-urea application into the rumen was given on days 1 (without glycerol infusion), 3 and 7 (with glycerol infusion). Extra samples of rumen fluid and blood plasma (from puncture of vena jungularis) were taken through the 12th hour following urea application. Rumen fluid pH was reduced due to glycerol intake from 6.3 (day 1, without glycerol) to 5.4 by day 7. Molar proportion of acetic acid to propionic acid decreased from 3.5 (day 1) to 2.1 (days 6 and 7). Average glycerol disappearance rate from the rumen was 4.7 gl-1 h-1 for the first hour. Only small amounts of glycerol could be detected in the duodenal digesta. Blood plasma glycerol content was significantly higher after glycerol application (0.061 mmol l-1 vs. 0.019 mmol l-1). The incorporation of 15N into the rumen bacteria and the proportion of bacterial N (as percent of TCA-precipitable N in the rumen fluid) were lower after glycerol influsion. These results, coupled with the lower concentration of iso-acids (isobutyric and isovaleric acids) in the rumen fluid, indicate that the high amount of glycerol infusion (10% of DMI) reduced protein metabolism of rumen bacteria throughout the experimental period.

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

    PubMed Central

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

    2015-01-01

    Aim: The present study was conducted to examine the rumen modulatory effect of thyme, clove and peppermint oils on rumen fermentation pattern in vitro using roughage based diet. Materials and Methods: Thyme, clove and peppermint oils were tested at concentration of 0, 30, 300 and 600 mg/l (ppm) of total culture fluid using in vitro gas production technique in wheat straw based diet (concentrate: Wheat straw 50:50). Different in vitro parameters e.g., total gas production, methane production, nutrient degradability, volatile fatty acid (VFA) production and ammonia nitrogen concentration were studied using buffalo rumen liquor. Results: Thyme oil at higher dose level (600 ppm) reduced (p<0.05) total gas production, feed degradability and ammonia nitrogen (NH3-N) concentration whereas total VFA concentration was significantly lower (p>0.05) in 300 and 600 ppm dose levels. 600 ppm dose level of clove oil reduced (p<0.05) total gas production, feed degradability, total VFA and acetate to propionate ratio. Methane production was significantly reduced (p<0.05) in 300 and 600 ppm dose levels of clove and peppermint oil. Conclusion: Right combination of these essential oils may prove to enhance performance of animals by reducing methane production and inhibiting protein degradation in rumen. PMID:27047073

  13. Manipulating rumen microbiome and fermentation through interventions during early life: a review

    PubMed Central

    Yáñez-Ruiz, David R.; Abecia, Leticia; Newbold, Charles J.

    2015-01-01

    The nutritional manipulations of the rumen microbiome to enhance productivity and health are rather limited by the resilience of the ecosystem once established in the mature rumen. Based on recent studies, it has been suggested that the microbial colonization that occurs soon after birth opens a possibility of manipulation with potential to produce lasting effects into adult life. This paper presents the state-of-the-art in relation to early life nutritional interventions by addressing three areas: the development of the rumen as an organ in regards to the nutrition of the new-born, the main factors that determine the microbial population that first colonizes and establishes in the rumen, and the key immunity players that contribute to shaping the commensal microbiota in the early stage of life to understand host-microbiome specificity. The development of the rumen epithelium and muscularization are differently affected by the nature of the diet and special care should be taken with regards to transition from liquid (milk) to solid feed. The rumen is quickly colonized by all type of microorganisms straight after birth and the colonization pattern may be influenced by several factors such as presence/absence of adult animals, the first solid diet provided, and the inclusion of compounds that prevent/facilitate the establishment of some microorganisms or the direct inoculation of specific strains. The results presented show how early life events may be related to the microbial community structure and/or the rumen activity in the animals post-weaning. This would create differences in adaptive capacity due to different early life experiences and leads to the idea of microbial programming. However, many elements need to be further studied such as: the most sensitive window of time for interventions, the best means to test long term effectiveness, the role of key microbial groups and host-immune regulations. PMID:26528276

  14. Manipulating rumen microbiome and fermentation through interventions during early life: a review.

    PubMed

    Yáñez-Ruiz, David R; Abecia, Leticia; Newbold, Charles J

    2015-01-01

    The nutritional manipulations of the rumen microbiome to enhance productivity and health are rather limited by the resilience of the ecosystem once established in the mature rumen. Based on recent studies, it has been suggested that the microbial colonization that occurs soon after birth opens a possibility of manipulation with potential to produce lasting effects into adult life. This paper presents the state-of-the-art in relation to early life nutritional interventions by addressing three areas: the development of the rumen as an organ in regards to the nutrition of the new-born, the main factors that determine the microbial population that first colonizes and establishes in the rumen, and the key immunity players that contribute to shaping the commensal microbiota in the early stage of life to understand host-microbiome specificity. The development of the rumen epithelium and muscularization are differently affected by the nature of the diet and special care should be taken with regards to transition from liquid (milk) to solid feed. The rumen is quickly colonized by all type of microorganisms straight after birth and the colonization pattern may be influenced by several factors such as presence/absence of adult animals, the first solid diet provided, and the inclusion of compounds that prevent/facilitate the establishment of some microorganisms or the direct inoculation of specific strains. The results presented show how early life events may be related to the microbial community structure and/or the rumen activity in the animals post-weaning. This would create differences in adaptive capacity due to different early life experiences and leads to the idea of microbial programming. However, many elements need to be further studied such as: the most sensitive window of time for interventions, the best means to test long term effectiveness, the role of key microbial groups and host-immune regulations.

  15. Effects of Eucalyptus Crude Oils Supplementation on Rumen Fermentation, Microorganism and Nutrient Digestibility in Swamp Buffaloes

    PubMed Central

    Thao, N. T.; Wanapat, M.; Cherdthong, A.; Kang, S.

    2014-01-01

    not affect nitrogen utilization. Both allantoin excretion and absorption and microbial nitrogen supply were increased by UTRS whereas efficiency of microbial protein synthesis was similar in all treatments. Findings of present study suggested that EuO could be used as a feed additive to modify the rumen fermentation in reducing methane production both in RS and UTRS. Feeding UTRS could improve feed intake and efficiency of rumen fermentation in swamp buffaloes. However, more research is warranted to determine the effect of EuO supplementation in production animals. PMID:25049925

  16. Effect of the type of silage on milk yield, intake and rumen metabolism of dairy cows grazing swards with low herbage mass.

    PubMed

    Ruiz-Albarrán, Miguel; Balocchi, Oscar A; Noro, Mirela; Wittwer, Fernando; Pulido, Rubén G

    2016-07-01

    The aim of this study was to evaluate the effect of herbage allowance (HA) and type of silage supplemented (TS) on milk yield, dry matter intake (DMI) and metabolism of dairy cows in early lactation. Thirty-six Holstein-Friesian dairy cows were allocated to four treatments derived from an arrangement of two HA (LHA = 17 or HHA = 25 kg of DM/cow/day) and two TS (grass (GS) or maize (MS)). Herbage allowance had no effect on DMI or milk yield. Rumen pH and NH3 -N concentration were not affected by HA. The efficiency of microbial protein synthesis in the rumen (microbial protein (MP)) was affected by HA with 21.5 and 23.9 g microbial nitrogen per kg ruminal digestible organic matter for LHA and HHA, respectively (P < 0.05). Supplementation with MS showed higher values of milk yield by 2.4 kg/cow/day (P < 0.001), milk protein content by 0.10 % (P < 0.023) and herbage DMI by 2.2 kg/cow/day, and showed lower values for milk urea compared to GS (P < 0.001). The former results suggest that TS had a greater effect on milk yield, total feed intake and energy intake than increase in herbage allowance; however, increase in HA had greater effects on MP than TS. © 2015 Japanese Society of Animal Science.

  17. Short communication: Intestinal digestibility of amino acids in fluid- and particle-associated rumen bacteria determined using a precision-fed cecectomized rooster bioassay.

    PubMed

    Fonseca, A C; Fredin, S M; Ferraretto, L F; Parsons, C M; Utterback, P L; Shaver, R D

    2014-01-01

    Microbial protein represents the majority of metabolizable protein absorbed by ruminant animals. Enhanced understanding of the AA digestibility of rumen microbes will improve estimates of metabolizable protein. The objective of this experiment was to determine the digestibility of AA in fluid- (FAB) and particle-associated bacteria (PAB) using the precision-fed cecectomized rooster bioassay. Bacteria were isolated from 4 ruminally cannulated lactating Holstein cows by differential centrifugation, including particle suspension in 0.1% Tween-80 for increased removal of PAB from ruminal digesta. Samples of FAB and PAB were fed to 9 cecectomized roosters to determine standardized digestibility of AA. Total AA digestibility was 76.8 and 75.5% for FAB and PAB, respectively, but did not differ. Differences existed in AA digestibilities within bacterial type when compared with the mean essential AA digestibility value. Compared with previous literature estimates of AA digestibility in microbes (mean = 76%; range = 57-87%) and relative to National Research Council estimates of total AA from rumen bacteria (80%), the precision-fed cecectomized rooster assay is an acceptable in vivo model to determine AA digestibility of rumen bacteria. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

    PubMed

    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.

  19. The effect of rumen ciliates on chitinolytic activity, chitin content and the number of fungal zoospores in the rumen fluid of sheep.

    PubMed

    Miltko, Renata; Bełżecki, Grzegorz; Herman, Andrzej; Kowalik, Barbara; Skomiał, Jacek

    2016-12-01

    The objective of this study was to investigate the effect of selected protozoa on the degradation and concentration of chitin and the numbers of fungal zoospores in the rumen fluid of sheep. Three adult ewes were fed a hay-concentrate diet, defaunated, then monofaunated with Entodinium caudatum or Diploplastron affine alone and refaunated with natural rumen fauna. The average density of the protozoa population varied from 6.1 · 10(4) (D. affine) to 42.2 · 10(4) cells/ml rumen fluid (natural rumen fauna). The inoculation of protozoa in the rumen of defaunated sheep increased the total activity of chitinolytic enzymes from 2.9 to 3.6 μmol N-acetylglucosamine/g dry matter (DM) of rumen fluid per min, the chitin concentration from 6.3 to 7.2 mg/g DM of rumen fluid and the number of fungal zoospores from 8.1 to 10.9 · 10(5) cells/ml rumen fluid. All examined indices showed diurnal variations. Ciliate population density was highest immediately prior to feeding and lowest at 4 h thereafter. The opposite effects were observed for the numbers of fungal zoospores, the chitin concentration and chitinolytic activity. Furthermore, it was found that chitin from zoospores may account for up to 95% of total microbial chitin in the rumen fluid of sheep. In summary, the examined ciliate species showed the ability of chitin degradation as well as a positive influence on the development of the ruminal fungal population.

  20. The effects of oral magnesium hydroxide administration on rumen fluid in cattle.

    PubMed

    Smith, Geoffrey W; Correa, Maria T

    2004-01-01

    This study was conducted to determine the effects of oral magnesium hydroxide administration on rumen fluid in cattle. Six lactating Holstein cows (4-7 years of age) with rumen fistulas were studied. Cattle were randomly assigned to receive boluses of magnesium hydroxide (162 g) or a powdered form (450 g dissolved in 3.5 L of water) PO daily for 3 days. Analysis of rumen fluid, blood gas tensions, and pH and measurement of serum magnesium concentrations were conducted daily. The study was discontinued after 72 hours, or sooner if rumen pH exceeded 8.0. After at least 3 weeks, the study was repeated with each cow receiving the other form of magnesium hydroxide (powder or bolus). Compared with baseline rumen pH (mean +/- SD: 6.22 +/- 0.28), magnesium hydroxide boluses caused a significant increase (P < .05) in rumen pH after 48 (7.27 +/- 0.11) and 72 (8.01 +/- 0.16) hours of administration, whereas the powdered form caused a significant increase (P < .05) in rumen pH after 24 (7.54 +/- 0.19) and 48 (8.43 +/- 0.22) hours of administration. Both the powdered and bolus forms of magnesium hydroxide decreased rumen protozoal numbers and increased methylene blue reduction times compared with baseline values. There was no change in blood pH, bicarbonate, or base excess values. Serum magnesium concentrations were significantly increased (P < .05) in cows that received the magnesium hydroxide powder. The results of this study indicate that magnesium hydroxide has a potent alkalinizing effect on rumen pH and significantly decreases rumen microbial activity.

  1. Effects of fish meals on rumen bacterial fermentation in continuous culture.

    PubMed

    Hoover, W H; Miller, T K; Stokes, S R; Thayne, W V

    1989-11-01

    Effects of various forms of fish meal on microbial metabolism were investigated in continuous cultures of rumen contents. Five diets were formulated to contain 12% ruminally degradable protein and 47 to 48% nonstructural carbohydrate. Soybean meal was the major protein source in the control diet, whereas in the other four diets, various fish meals were substituted for 6% of total diet DM. Fish meals were: fish meal containing 34.4% FFA, fish meal containing 34.4% FFA with CaCl2 added, fish meal containing 65.6% FFA, and fish meal defatted using 1:1 ethanol:ether extraction. The five treatments were fermented with pH either held constant at 6.2 or not controlled. When pH was maintained at 6.2, the inclusion of any fish meal except defatted fish meal reduced the acetate:propionate ratio, decreased protein digestion, and reduced microbial N produced/per kilogram DM digested when compared with the soybean control. When not controlled, pH decreased after feeding to 6.0 or lower. Under these conditions, the soybean control had a lower acetate:propionate ratio and lower NDF digestion than all diets containing fish meal. In this study, oil-containing fish meal affected microbial metabolism more negatively when the fermentation pH was held at 6.2 than when the pH was 6.0 or lower.

  2. Genome sequencing of rumen bacteria and archaea and its application to methane mitigation strategies.

    PubMed

    Leahy, S C; Kelly, W J; Ronimus, R S; Wedlock, N; Altermann, E; Attwood, G T

    2013-06-01

    Ruminant-derived methane (CH4), a potent greenhouse gas, is a consequence of microbial fermentation in the digestive tract of livestock. Development of mitigation strategies to reduce CH4 emissions from farmed animals is currently the subject of both scientific and environmental interest. Methanogens are the sole producers of ruminant CH4, and therefore CH4 abatement strategies can either target the methanogens themselves or target the other members of the rumen microbial community that produce substrates necessary for methanogenesis. Understanding the relationship that methanogens have with other rumen microbes is crucial when considering CH4 mitigation strategies for ruminant livestock. Genome sequencing of rumen microbes is an important tool to improve our knowledge of the processes that underpin those relationships. Currently, several rumen bacterial and archaeal genome projects are either complete or underway. Genome sequencing is providing information directly applicable to CH4 mitigation strategies based on vaccine and small molecule inhibitor approaches. In addition, genome sequencing is contributing information relevant to other CH4 mitigation strategies. These include the selection and breeding of low CH4-emitting animals through the interpretation of large-scale DNA and RNA sequencing studies and the modification of other microbial groups within the rumen, thereby changing the dynamics of microbial fermentation.

  3. Effect of administration of rumen fungi on production performance of lactating buffaloes.

    PubMed

    Saxena, S; Sehgal, J P; Puniya, A K; Singh, K

    2010-06-01

    Anaerobic fungi were orally dosed to lactating buffaloes to study their effect on the digestibility of a diet (composed of 50% wheat straw and 50% concentrate along with six kg maize green/animal/day), rumen fermentation patterns and milk production. Group I (control) was administered with fungus-free anaerobic broth, while group II and III were administered with Orpinomyces sp. C-14 or Piromyces sp. WNG-12 (250 ml; 3-5 days of growth/animal/ week), respectively. Milk production was higher in group II and III (8.42 and 8.48 kg/d) than in the control (8.03 kg/d) with virtually the same feed intake (i.e. 11.50 and 10.62 and 11.79 kg, respectively). There was an increase of 6% fat-corrected milk yield/animal/day in group II and III, respectively compared to the control. The milk fat was higher in the fungal culture administered groups than in the control group. The digestibility of dry matter, crude protein, neutral detergent fibre, acid detergent fibre, cellulose and digestible energy also increased significantly in group II and III. The pH and ammonia nitrogen were lower, whereas total volatile fatty acids, total nitrogen, trichloroacid precipitable nitrogen and number of zoospores/ml of rumen liquor were higher in group II and III when compared to the control. Hence, it can be stated that rumen fungi can be used as a direct-fed microbial in lactating buffaloes, to enhance the digestibility of wheat straw based diets leading to higher production.

  4. Insights from quantitative metaproteomics and protein-stable isotope probing into microbial ecology.

    PubMed

    von Bergen, Martin; Jehmlich, Nico; Taubert, Martin; Vogt, Carsten; Bastida, Felipe; Herbst, Florian-Alexander; Schmidt, Frank; Richnow, Hans-Hermann; Seifert, Jana

    2013-10-01

    The recent development of metaproteomics has enabled the direct identification and quantification of expressed proteins from microbial communities in situ, without the need for microbial enrichment. This became possible by (1) significant increases in quality and quantity of metagenome data and by improvements of (2) accuracy and (3) sensitivity of modern mass spectrometers (MS). The identification of physiologically relevant enzymes can help to understand the role of specific species within a community or an ecological niche. Beside identification, relative and absolute quantitation is also crucial. We will review label-free and label-based methods of quantitation in MS-based proteome analysis and the contribution of quantitative proteome data to microbial ecology. Additionally, approaches of protein-based stable isotope probing (protein-SIP) for deciphering community structures are reviewed. Information on the species-specific metabolic activity can be obtained when substrates or nutrients are labeled with stable isotopes in a protein-SIP approach. The stable isotopes ((13)C, (15)N, (36)S) are incorporated into proteins and the rate of incorporation can be used for assessing the metabolic activity of the corresponding species. We will focus on the relevance of the metabolic and phylogenetic information retrieved with protein-SIP studies and for detecting and quantifying the carbon flux within microbial consortia. Furthermore, the combination of protein-SIP with established tools in microbial ecology such as other stable isotope probing techniques are discussed.

  5. In vitro study (Rusitec) of the action of abierixin, a new ionophore antibiotic, on the end products of fermentation and the degradation of nitrogen in the rumen.

    PubMed

    Gomez, L; Hillaire, M C; Jouany, J P

    1990-03-01

    We investigated the capacity of a new ionophore antibiotic, abierixin, to modify fermentations in the rumen using a semi-continuous fermenter (Rusitec). As in the studies carried out on a "batch" fermenter (HILLAIRE et al., 1989a), abierixin failed to alter volatile fatty acids and gas productions but, in contrast, it limited the degradation of dietary nitrogen without affecting microbial synthesis. This molecule which has a low level of toxicity and is capable of improving the use of dietary proteins by ruminants, was found to be more effective when used at very low dose levels.

  6. Molecular cloning and characterization of a novel SGNH arylesterase from the goat rumen contents.

    PubMed

    Wang, Guozeng; Meng, Kun; Luo, Huiying; Wang, Yaru; Huang, Huoqing; Shi, Pengjun; Pan, Xia; Yang, Peilong; Yao, Bin

    2011-09-01

    An esterase-encoding gene, estR5, was directly obtained from the genomic DNA of goat rumen contents. The 555-bp full-length gene encodes a 184-residue polypeptide (EstR5) without putative signal peptide. Deduced EstR5 shared the highest identity (50%) to a putative arylesterase from Ruminococcaceae bacterium D16. Phylogenetic analysis indicated that EstR5 was closely related with microbial esterases of gastrointestinal source. A comparison of the conserved motifs shared with GDSL proteins revealed that EstR5 could be grouped into the GDSL family and was further classified into the subfamily of SGNH hydrolases. The gene estR5 was expressed in Escherichia coli BL21 (DE3) and purified to electrophoretic homogeneity. Recombinant EstR5 exhibited highest catalytic efficiency towards α-naphthyl acetate followed by phenyl acetate and p-nitrophenyl acetate and had no activity towards PNP esters with acyl chains longer than C8. The enzyme exhibited optimal activity at around 60°C and pH 8.0, was stable at pH ranging from 6.0 to 11.0 and was slightly activated by detergent Tween, Nonidet P-40, and Triton X-100. These properties suggest that EstR5 has great potential for basic research and industrial applications. To our knowledge, this is the first arylesterase obtained from rumen microenvironment.

  7. High-throughput Methods Redefine the Rumen Microbiome and Its Relationship with Nutrition and Metabolism

    PubMed Central

    McCann, Joshua C.; Wickersham, Tryon A.; Loor, Juan J.

    2014-01-01

    Diversity in the forestomach microbiome is one of the key features of ruminant animals. The diverse microbial community adapts to a wide array of dietary feedstuffs and management strategies. Understanding rumen microbiome composition, adaptation, and function has global implications ranging from climatology to applied animal production. Classical knowledge of rumen microbiology was based on anaerobic, culture-dependent methods. Next-generation sequencing and other molecular techniques have uncovered novel features of the rumen microbiome. For instance, pyrosequencing of the 16S ribosomal RNA gene has revealed the taxonomic identity of bacteria and archaea to the genus level, and when complemented with barcoding adds multiple samples to a single run. Whole genome shotgun sequencing generates true metagenomic sequences to predict the functional capability of a microbiome, and can also be used to construct genomes of isolated organisms. Integration of high-throughput data describing the rumen microbiome with classic fermentation and animal performance parameters has produced meaningful advances and opened additional areas for study. In this review, we highlight recent studies of the rumen microbiome in the context of cattle production focusing on nutrition, rumen development, animal efficiency, and microbial function. PMID:24940050

  8. Metatranscriptomic Profiling Reveals Linkages between the Active Rumen Microbiome and Feed Efficiency in Beef Cattle.

    PubMed

    Li, Fuyong; Guan, Le Luo

    2017-05-01

    Exploring compositional and functional characteristics of the rumen microbiome can improve the understanding of its role in rumen function and cattle feed efficiency. In this study, we applied metatranscriptomics to characterize the active rumen microbiomes of beef cattle with different feed efficiencies (efficient, n = 10; inefficient, n = 10) using total RNA sequencing. Active bacterial and archaeal compositions were estimated based on 16S rRNAs, and active microbial metabolic functions including carbohydrate-active enzymes (CAZymes) were assessed based on mRNAs from the same metatranscriptomic data sets. In total, six bacterial phyla (Proteobacteria, Firmicutes, Bacteroidetes, Spirochaetes, Cyanobacteria, and Synergistetes), eight bacterial families (Succinivibrionaceae, Prevotellaceae, Ruminococcaceae, Lachnospiraceae, Veillonellaceae, Spirochaetaceae, Dethiosulfovibrionaceae, and Mogibacteriaceae), four archaeal clades (Methanomassiliicoccales, Methanobrevibacter ruminantium, Methanobrevibacter gottschalkii, and Methanosphaera), 112 metabolic pathways, and 126 CAZymes were identified as core components of the active rumen microbiome. As determined by comparative analysis, three bacterial families (Lachnospiraceae, Lactobacillaceae, and Veillonellaceae) tended to be more abundant in low-feed-efficiency (inefficient) animals (P < 0.10), and one archaeal taxon (Methanomassiliicoccales) tended to be more abundant in high-feed-efficiency (efficient) cattle (P < 0.10). Meanwhile, 32 microbial metabolic pathways and 12 CAZymes were differentially abundant (linear discriminant analysis score of >2 with a P value of <0.05) between two groups. Among them, 30 metabolic pathways and 11 CAZymes were more abundant in the rumen of inefficient cattle, while 2 metabolic pathways and 1 CAZyme were more abundant in efficient animals. These findings suggest that the rumen microbiomes of inefficient cattle have more diverse activities than those of efficient cattle, which may be

  9. Comparative efficiency of microbial enzyme preparations versus pancreatin for in vitro alimentary protein digestion.

    PubMed

    Andriamihaja, Mireille; Guillot, Alain; Svendsen, Allan; Hagedorn, Joerg; Rakotondratohanina, Sahondra; Tomé, Daniel; Blachier, François

    2013-02-01

    Utilisation of microbial enzymes may represent an alternative strategy to the use of conventional pancreatin obtained from pig pancreas for the treatment of severe pancreatic insufficiency. In this study, we focused on the capacity of two microbial preparations for their capacity to digest alimentary proteins (caseins and soya proteins) in comparison with pancreatin. These microbial enzymatic preparations were found to be able to generate small, medium-size and larger polypeptides from caseins and soya proteins but were inactivated at pH 3.0. As determined by Liquid Chromatography-Mass Spectrometry analysis, microbial enzymes generated very different peptides from caseins when compared with peptides generated through pancreatin action. These microbial preparations were characterised by relatively low trypsin- and low carboxypeptidase-like activities but high chymotrypsin-like activities and strong capacity for cleavage of caseins at the methionine sites. Although the efficiency of these microbial preparations to increase the rate of absorption of nitrogen-containing compounds in severe pancreatic insufficiency remains to be tested in vivo, our in vitro data indicate proteolytic capacities of such preparations for alimentary protein digestion.

  10. Effects of addition of Aspergillus oryzae culture and 2-hydroxyl-4-(methylthio) butanoic acid on milk performance and rumen fermentation of dairy cows.

    PubMed

    Sun, Hua; Wu, Yueming; Wang, Yanming; Wang, Chong; Liu, Jianxin

    2017-04-01

    To investigate effects of Aspergillus oryzae culture (AOC) and 2-hydroxy-4-(methylthio) butanoic acid (HMB) on milk performance and rumen fermentation of dairy cows. Sixty-four multiparous Chinese Holstein cows were randomly allocated into four experimental diets: (i) Control diet; (ii) AOC diet: 5 g AOC/day per head; (iii) HMB diet: 25 g HMB/day; and (iv) AH diet: 5 g AOC plus 25 g HMB/day. Added HMB tended to increase the yield of milk protein (P = 0.06) and 3.5% fat-corrected milk (P = 0.08) and milk fat content (P = 0.09). Milk fat yield (P = 0.03) and the contents of milk protein (P = 0.05) were increased by adding HMB. The cows fed on AOC diet had a tendency for higher body weight (BW) gain (P = 0.08). Addition of AOC, HMB and AH increased content of microbial protein (MCP) and total volatile fatty acids (VFA) (P < 0.01) in rumen fluid. Populations of rumen fungi, Fibrobacter succinogenes and Ruminococcus flavefaciens relative to total bacterial 16S rDNA (P ≤ 0.03) and activity of carboxymethylcellulase (CMCase) (P < 0.01) were increased with added AOC or HMB. It is inferred that added AOC or HMB can increase the contents of MCP and total VFA potentially by stimulating rumen microbe populations and CMCase activity. © 2016 Japanese Society of Animal Science.

  11. Impairment of rumen biohydrogenation and bacteria of the Butyrivibrio group in the rumen of goats through a 20:5 n-3 (EPA) rich supplement.

    PubMed

    Lv, Xuejiao; Mao, Shengyong; Zhu, Weiyun

    2016-01-30

    Marine products can inhibit biohydrogenation in the rumen, but the mechanism is not clear. This study investigated a 20:5 n-3 rich supplement effects on rumen biohydrogenation, microbial change and fermentation characteristics in goats. The supplementation decreased 18:0 proportions in rumen fatty acids (P < 0.001), while it increased cis-9, trans-11 conjugated linoleic acid (CLA) (P < 0.001) and trans-10, cis-12 CLA proportions (P < 0.001). The supplement reduced the number of Butyrivibrio spp. and B. proteoclasticus (P < 0.01). Denaturing gradient gel electrophoresis redundancy analysis indicated that some species, mainly from the rumen of goats receiving the 2.5 and 5.0 g d(-1) supplement, were positively correlated with cis-9, trans-11 CLA proportions; some species, mainly from the rumen of control goats, were positively correlated with 18:0 proportions. The supplement reduced the NH3 -N concentrations and acetate molar proportions in the rumen (P < 0.05), but increased propionate and butyrate molar proportions (P < 0.01), and had no effect on total volatile fatty acid concentration. The supplement rich in 20:5 n-3 reduced the biohydrogenation of 18-carbon unsaturated fatty acids with a significant reduction of the 18:0 proportion and this was coupled with the suppression of the abundance of biohydrogenating bacteria and unknown bacteria. © 2015 Society of Chemical Industry.

  12. Redirection of Metabolic Hydrogen by Inhibiting Methanogenesis in the Rumen Simulation Technique (RUSITEC)

    PubMed Central

    Guyader, Jessie; Ungerfeld, Emilio M.; Beauchemin, Karen A.

    2017-01-01

    A decrease in methanogenesis is expected to improve ruminant performance by allocating rumen metabolic hydrogen ([2H]) to more energy-rendering fermentation pathways for the animal. However, decreases in methane (CH4) emissions of up to 30% are not always linked with greater performance. Therefore, the aim of this study was to understand the fate of [2H] when CH4 production in the rumen is inhibited by known methanogenesis inhibitors (nitrate, NIT; 3-nitrooxypropanol, NOP; anthraquinone, AQ) in comparison with a control treatment (CON) with the Rumen Simulation Technique (RUSITEC). Measurements started after 1 week adaptation. Substrate disappearance was not modified by methanogenesis inhibitors. Nitrate mostly seemed to decrease [2H] availability by acting as an electron acceptor competing with methanogenesis. As a consequence, NIT decreased CH4 production (−75%), dissolved dihydrogen (H2) concentration (−30%) and the percentages of reduced volatile fatty acids (butyrate, isobutyrate, valerate, isovalerate, caproate and heptanoate) except propionate, but increased acetate molar percentage, ethanol concentration and the efficiency of microbial nitrogen synthesis (+14%) without affecting gaseous H2. Nitrooxypropanol decreased methanogenesis (−75%) while increasing both gaseous and dissolved H2 concentrations (+81% and +24%, respectively). Moreover, NOP decreased acetate and isovalerate molar percentages and increased butyrate, valerate, caproate and heptanoate molar percentages as well as n-propanol and ammonium concentrations. Methanogenesis inhibition with AQ (−26%) was associated with higher gaseous H2 production (+70%) but lower dissolved H2 concentration (−76%), evidencing a lack of relationship between the two H2 forms. Anthraquinone increased ammonium concentration, caproate and heptanoate molar percentages but decreased acetate and isobutyrate molar percentages, total microbial nitrogen production and efficiency of microbial protein synthesis (

  13. Effect of bromochloromethane on methane emission, rumen fermentation pattern, milk yield, and fatty acid profile in lactating dairy goats.

    PubMed

    Abecia, L; Toral, P G; Martín-García, A I; Martínez, G; Tomkins, N W; Molina-Alcaide, E; Newbold, C J; Yáñez-Ruiz, D R

    2012-04-01

    Several technologies have been tested to reduce enteric methanogenesis, but very few have been successfully used in practical conditions for livestock. Furthermore, the consequences of reduced rumen methane production on animal performance and milk quality are poorly understood. The aim of this work was to investigate the effect of feeding bromochloromethane (BCM), a halogenated aliphatic hydrocarbon with potential antimethanogenic activity, to dairy goats on rumen methane production, fermentation pattern, the abundance of major microbial groups, and on animal performance and milk composition. Eighteen goats were allocated to 2 experimental groups of 9 animals each: treated (BCM+) or not (BCM-) with 0.30 g of BCM/100 kg of body weight per day. The BCM was administered per os in 2 equal doses per day from parturition to 2 wk postweaning (10 wk). After weaning, methane emissions were recorded over 2 consecutive days (d 57 and 58 on treatment) in polycarbonate chambers. On d 59, individual rumen fluid samples were collected for volatile fatty acid (VFA) analysis and quantification of bacterial, protozoal, and archaeal numbers by real-time PCR. On d 69 and 70, daily milk production was recorded and samples were collected for determination of fat, protein, lactose, casein, and total solids concentration by infrared spectrophotometry, and fatty acid composition by gas chromatography. Treatment with BCM reduced methane production by 33% (21.6 vs. 14.4 L/kg of DMI) compared with nontreated animals, although it did not affect the abundance of rumen bacteria, protozoa, and total methanogenic archaea. The observed improvement in the efficiency of digestive processes was accompanied by a 36% increase in milk yield, probably due to the more propionic type of rumen fermentation and an increase in VFA production. The increase in milk yield was not accompanied by any changes in the concentrations or yields of fat, protein, or lactose. Despite the substantial decrease in methane

  14. Invited review: adhesion mechanisms of rumen cellulolytic bacteria.

    PubMed

    Miron, J; Ben-Ghedalia, D; Morrison, M

    2001-06-01

    We divided the adhesion process of the predominant cellulolytic rumen bacteria Fibrobacter succinogenes, Ruminococcus flavefaciens, and Ruminococcus albus into four phases: 1) transport of the nonmotile bacteria to the substrate; 2) initial nonspecific adhesion of bacteria to unprotected sites of the substrate that is dominated by constitutive elements of bacterial glycocalyx; 3) specific adhesion via adhesins or ligands formation with the substrate, which can be dominated by several bacterial organelles including cellulosome complexes, fimbriae connections, glycosylated epitopes of cellulose-binding protein (CBP) or glycocalyx, and cellulose-binding domain (CBD) of enzymes; 4) proliferation of the attached bacteria on potentially digestible tissues of the substrate. Each of the phases and its significance in the adhesion process are described. Factors affecting bacterial adhesion are described including: 1) factors related to bacterial age, glycocalyx condition, and microbial competition; 2) factors related to the nature of substrate including, cuticle protection, surface area, hydration, and ionic charge; and 3) environmental factors including pH, temperature, and presence of cations and soluble carbohydrate. Based on the information available from the literature, it appears that each of the predominant rumen bacteria--F. succinogenes, R. flavefaciens, and R. albus--has a specific mechanism of adhesion to cellulose. In F. succinogenes, both the glycosidic residues of the outer membrane CBP and especially of the 180-kDa CBP, and the distinct CBD of EG2 EGF and Cl-stimulated cellobiosidase, may play a role in the adhesion to cellulose. No direct evidence, except scanning electron microscopy observations, yet supports the existence of either cellulosome complex or fimbriae structures involved in the adhesion mechanism of F. succinogenes. At least two mechanisms, cellulosome-like complexes and carbohydrate epitopes of the glycocalyx layer are involved in the specific

  15. Effects of corn and soybean meal types on rumen fermentation, nitrogen metabolism and productivity in dairy cows.

    PubMed

    Shen, J S; Song, L J; Sun, H Z; Wang, B; Chai, Z; Chacher, B; Liu, J X

    2015-03-01

    Twelve multiparous Holstein dairy cows in mid-lactation were selected for a replicated 4×4 Latin square design with a 2 ×2 factorial arrangement to investigate the effects of corn and soybean meal (SBM) types on rumen fermentation, N metabolism and lactation performance in dairy cows. Two types of corn (dry ground [DGC] and steam-flaked corn [SFC]) and two types of SBM (solvent-extracted and heat-treated SBM) with different ruminal degradation rates and extents were used to formulate four diets with the same basal ingredients. Each period lasted for 21 days, including 14 d for adaptation and 7 d for sample collection. Cows receiving SFC had a lower dry matter (DM) and total N intake than those fed DGC. However, the milk yield and milk protein yield were not influenced by the corn type, resulting in higher feed and N utilization efficiency in SFC-fed cows than those receiving DGC. Ruminal acetate concentrations was greater and total volatile fatty acids concentrations tended to be greater for cows receiving DGC relative to cows fed SFC, but milk fat content was not influenced by corn type. The SFC-fed cows had lower ruminal ammonia-N, less urea N in their blood and milk, and lower fecal N excretion than those on DGC. Compared with solvent-extracted SBM-fed cows, cows receiving heat-treated SBM had lower microbial protein yield in the rumen, but similar total tract apparent nutrient digestibility, N metabolism measurements, and productivity. Excessive supply of metabolizable protein in all diets may have caused the lack of difference in lactation performance between SBM types. Results of the present study indicated that increasing the energy degradability in the rumen could improve feed efficiency, and reduce environmental pollution.

  16. Effects of Corn and Soybean Meal Types on Rumen Fermentation, Nitrogen Metabolism and Productivity in Dairy Cows

    PubMed Central

    Shen, J. S.; Song, L. J.; Sun, H. Z.; Wang, B.; Chai, Z.; Chacher, B.; Liu, J. X.

    2015-01-01

    Twelve multiparous Holstein dairy cows in mid-lactation were selected for a replicated 4×4 Latin square design with a 2 ×2 factorial arrangement to investigate the effects of corn and soybean meal (SBM) types on rumen fermentation, N metabolism and lactation performance in dairy cows. Two types of corn (dry ground [DGC] and steam-flaked corn [SFC]) and two types of SBM (solvent-extracted and heat-treated SBM) with different ruminal degradation rates and extents were used to formulate four diets with the same basal ingredients. Each period lasted for 21 days, including 14 d for adaptation and 7 d for sample collection. Cows receiving SFC had a lower dry matter (DM) and total N intake than those fed DGC. However, the milk yield and milk protein yield were not influenced by the corn type, resulting in higher feed and N utilization efficiency in SFC-fed cows than those receiving DGC. Ruminal acetate concentrations was greater and total volatile fatty acids concentrations tended to be greater for cows receiving DGC relative to cows fed SFC, but milk fat content was not influenced by corn type. The SFC-fed cows had lower ruminal ammonia-N, less urea N in their blood and milk, and lower fecal N excretion than those on DGC. Compared with solvent-extracted SBM-fed cows, cows receiving heat-treated SBM had lower microbial protein yield in the rumen, but similar total tract apparent nutrient digestibility, N metabolism measurements, and productivity. Excessive supply of metabolizable protein in all diets may have caused the lack of difference in lactation performance between SBM types. Results of the present study indicated that increasing the energy degradability in the rumen could improve feed efficiency, and reduce environmental pollution. PMID:25656206

  17. A Metagenomics Approach to Evaluate the Impact of Dietary Supplementation with Ascophyllum nodosum or Laminaria digitata on Rumen Function in Rusitec Fermenters.

    PubMed

    Belanche, Alejandro; Jones, Eleanor; Parveen, Ifat; Newbold, Charles J

    2016-01-01

    There is an increasing need to identify alternative feeds for livestock that do not compete with foods for humans. Seaweed might provide such a resource, but there is limited information available on its value as an animal feed. Here we use a multi-omics approach to investigate the value of two brown seaweeds, Ascophyllum nodosum (ASC) and Laminaria digitata (LAM), as alternative feeds for ruminants. These seaweeds were supplemented at 5% inclusion rate into a control diet (CON) in a rumen simulation fermenter. The seaweeds had no substantial effect on rumen fermentation, feed degradability or methane emissions. Concentrations of total bacteria, anaerobic fungi, biodiversity indices and abundances of the main bacterial and methanogen genera were also unaffected. However, species-specific effects of brown seaweed on the rumen function were noted: ASC promoted a substantial decrease in N degradability (-24%) due to its high phlorotannins content. Canonical correspondence analysis of the bacterial community revealed that low N availability led to a change in the structure of the bacterial community. ASC also decreased the concentration of Escherichia coli O157:H7 post-inoculation. In contrast, LAM which has a much lower phlorotannin content did not cause detrimental effects on N degradability nor modified the structure of the bacterial community in comparison to CON. This adaptation of the microbial community to LAM diets led to a greater microbial ability to digest xylan (+70%) and carboxy-methyl-cellulose (+41%). These differences among brown seaweeds resulted in greater microbial protein synthesis (+15%) and non-ammonia N flow (+11%) in LAM than in ASC diets and thus should led to a greater amino acid supply to the intestine of the animal. In conclusion, it was demonstrated that incorporation of brown seaweed into the diet can be considered as a suitable nutritional strategy for ruminants; however, special care must be taken with those seaweeds with high

  18. A Metagenomics Approach to Evaluate the Impact of Dietary Supplementation with Ascophyllum nodosum or Laminaria digitata on Rumen Function in Rusitec Fermenters

    PubMed Central

    Belanche, Alejandro; Jones, Eleanor; Parveen, Ifat; Newbold, Charles J.

    2016-01-01

    There is an increasing need to identify alternative feeds for livestock that do not compete with foods for humans. Seaweed might provide such a resource, but there is limited information available on its value as an animal feed. Here we use a multi-omics approach to investigate the value of two brown seaweeds, Ascophyllum nodosum (ASC) and Laminaria digitata (LAM), as alternative feeds for ruminants. These seaweeds were supplemented at 5% inclusion rate into a control diet (CON) in a rumen simulation fermenter. The seaweeds had no substantial effect on rumen fermentation, feed degradability or methane emissions. Concentrations of total bacteria, anaerobic fungi, biodiversity indices and abundances of the main bacterial and methanogen genera were also unaffected. However, species-specific effects of brown seaweed on the rumen function were noted: ASC promoted a substantial decrease in N degradability (−24%) due to its high phlorotannins content. Canonical correspondence analysis of the bacterial community revealed that low N availability led to a change in the structure of the bacterial community. ASC also decreased the concentration of Escherichia coli O157:H7 post-inoculation. In contrast, LAM which has a much lower phlorotannin content did not cause detrimental effects on N degradability nor modified the structure of the bacterial community in comparison to CON. This adaptation of the microbial community to LAM diets led to a greater microbial ability to digest xylan (+70%) and carboxy-methyl-cellulose (+41%). These differences among brown seaweeds resulted in greater microbial protein synthesis (+15%) and non-ammonia N flow (+11%) in LAM than in ASC diets and thus should led to a greater amino acid supply to the intestine of the animal. In conclusion, it was demonstrated that incorporation of brown seaweed into the diet can be considered as a suitable nutritional strategy for ruminants; however, special care must be taken with those seaweeds with high

  19. Rumen Bacterial Community Composition in Holstein and Jersey Cows Is Different under Same Dietary Condition and Is Not Affected by Sampling Method.

    PubMed

    Paz, Henry A; Anderson, Christopher L; Muller, Makala J; Kononoff, Paul J; Fernando, Samodha C

    2016-01-01

    The rumen microbial community in dairy cows plays a critical role in efficient milk production. However, there is a lack of data comparing the composition of the rumen bacterial community of the main dairy breeds. This study utilizes 16S rRNA gene sequencing to describe the rumen bacterial community composition in Holstein and Jersey cows fed the same diet by sampling the rumen microbiota via the rumen cannula (Holstein cows) or esophageal tubing (both Holstein and Jersey cows). After collection of the rumen sample via esophageal tubing, particles attached to the strainer were added to the sample to ensure representative sampling of both the liquid and solid fraction of the rumen contents. Alpha diversity metrics, Chao1 and observed OTUs estimates, displayed higher (P = 0.02) bacterial richness in Holstein compared to Jersey cows and no difference (P > 0.70) in bacterial community richness due to sampling method. The principal coordinate analysis displayed distinct clustering of bacterial communities by breed suggesting that Holstein and Jersey cows harbor different rumen bacterial communities. Family level classification of most abundant (>1%) differential OTUs displayed that OTUs from the bacterial families Lachnospiraceae and p-2534-18B5 to be predominant in Holstein cows compared to Jersey cows. Additionally, OTUs belonging to family Prevotellaceae were differentially abundant in the two breeds. Overall, the results from this study suggest that the bacterial community between Holstein and Jersey cows differ and that esophageal tubing with collection of feed particles associated with the strainer provides a representative rumen sample similar to a sample collected via the rumen cannula. Thus, in future studies esophageal tubing with addition of retained particles can be used to collect rumen samples reducing the cost of cannulation and increasing the number of animals used in microbiome investigations, thus increasing the statistical power of rumen microbial

  20. Rumen Bacterial Community Composition in Holstein and Jersey Cows Is Different under Same Dietary Condition and Is Not Affected by Sampling Method

    PubMed Central

    Paz, Henry A.; Anderson, Christopher L.; Muller, Makala J.; Kononoff, Paul J.; Fernando, Samodha C.

    2016-01-01

    The rumen microbial community in dairy cows plays a critical role in efficient milk production. However, there is a lack of data comparing the composition of the rumen bacterial community of the main dairy breeds. This study utilizes 16S rRNA gene sequencing to describe the rumen bacterial community composition in Holstein and Jersey cows fed the same diet by sampling the rumen microbiota via the rumen cannula (Holstein cows) or esophageal tubing (both Holstein and Jersey cows). After collection of the rumen sample via esophageal tubing, particles attached to the strainer were added to the sample to ensure representative sampling of both the liquid and solid fraction of the rumen contents. Alpha diversity metrics, Chao1 and observed OTUs estimates, displayed higher (P = 0.02) bacterial richness in Holstein compared to Jersey cows and no difference (P > 0.70) in bacterial community richness due to sampling method. The principal coordinate analysis displayed distinct clustering of bacterial communities by breed suggesting that Holstein and Jersey cows harbor different rumen bacterial communities. Family level classification of most abundant (>1%) differential OTUs displayed that OTUs from the bacterial families Lachnospiraceae and p-2534-18B5 to be predominant in Holstein cows compared to Jersey cows. Additionally, OTUs belonging to family Prevotellaceae were differentially abundant in the two breeds. Overall, the results from this study suggest that the bacterial community between Holstein and Jersey cows differ and that esophageal tubing with collection of feed particles associated with the strainer provides a representative rumen sample similar to a sample collected via the rumen cannula. Thus, in future studies esophageal tubing with addition of retained particles can be used to collect rumen samples reducing the cost of cannulation and increasing the number of animals used in microbiome investigations, thus increasing the statistical power of rumen microbial

  1. Development of microbial spoilage and lipid and protein oxidation in rabbit meat.

    PubMed

    Nakyinsige, K; Sazili, A Q; Aghwan, Z A; Zulkifli, I; Goh, Y M; Abu Bakar, F; Sarah, S A

    2015-10-01

    This experiment aimed to determine microbial spoilage and lipid and protein oxidation during aerobic refrigerated (4°C) storage of rabbit meat. Forty male New Zealand white rabbits were slaughtered according to the Halal slaughter procedure. The hind limbs were used for microbial analysis while the Longissimus lumborum m. was used for determination of lipid and protein oxidation. Bacterial counts generally increased with aging time and the limit for fresh meat (10(8)cfu/g) was reached at d 7 postmortem. Significant differences in malondialdehyde content were observed after 3d of storage. The thiol concentration significantly decreased with increase in aging time. The band intensities of myosin heavy chain and troponin T significantly reduced with increased refrigerated storage while actin remained relatively stable. This study thus proposes protein oxidation as a potential deteriorative change in refrigerated rabbit meat along with microbial spoilage and lipid oxidation.

  2. Interaction of unsaturated fat or coconut oil with monensin in lactating dairy cows fed 12 times daily. I. Protozoal abundance, nutrient digestibility, and microbial protein flow to the omasum.

    PubMed

    Reveneau, C; Karnati, S K R; Oelker, E R; Firkins, J L

    2012-04-01

    Monensin (tradename: Rumensin) should reduce the extent of amino acid deamination in the rumen, and supplemental fat should decrease protozoal abundance and intraruminal N recycling. Because animal-vegetable (AV) fat can be biohydrogenated in the rumen and decrease its effectiveness as an anti-protozoal agent, we included diets supplemented with coconut oil (CNO) to inhibit protozoa. In a 6 × 6 Latin square design with a 2 × 3 factorial arrangement of treatments, 6 rumen-cannulated cows were fed diets without or with Rumensin (12 g/909 kg) and either no fat (control), 5% AV fat, or 5% CNO. The log10 concentrations (cells/mL) of total protozoa were not different between control (5.97) and AV fat (5.95) but were decreased by CNO (4.79; main effect of fat source). Entodinium and Dasytricha decreased as a proportion of total cells from feeding CNO, whereas Epidinium was unchanged in total abundance and thus increased proportionately. Total volatile fatty acid concentration was not affected by diet, but the acetate:propionate ratio decreased for CNO (1.85) versus control (2.95) or AV fat (2.58). Feeding CNO (23.8%) decreased ruminal neutral detergent fiber digestibility compared with control (31.1%) and AV fat (30.5%). The total-tract digestibility of NDF was lower for CNO (45.8%) versus control (57.0%) and AV fat (54.6%), with no difference in apparent organic matter digestibility (averaging 69.8%). The omasal flows of microbial N and non-ammonia N were lower for CNO versus control and AV fat, but efficiency of microbial protein synthesis was not affected. The dry matter intake was 4.5 kg/d lower with CNO, which decreased milk production by 3.1 kg/d. Main effect means of dry matter intake and milk yield tended to decrease by 0.7 and 1.2 kg/d, respectively, when Rumensin was added. Both percentage and production of milk fat decreased for CNO (main effect of fat source). An interaction was observed such that AV decreased milk fat yield more when combined with Rumensin

  3. Determination of Microbial Growth by Protein Assay in an Air-Cathode Single Chamber Microbial Fuel Cell.

    PubMed

    Li, Na; Kakarla, Ramesh; Moon, Jung Mi; Min, Booki

    2015-07-01

    Microbial fuel cells (MFCs) have gathered attention as a novel bioenergy technology to simultaneously treat wastewater with less sludge production than the conventional activated sludge system. In two different operations of the MFC and aerobic process, microbial growth was determined by the protein assay method and their biomass yields using real wastewater were compared. The biomass yield on the anode electrode of the MFC was 0.02 g-COD-cell/g- COD-substrate and the anolyte planktonic biomass was 0.14 g-COD-cell/g-COD-substrate. An MFC without anode electrode resulted in the biomass yield of 0.07 ± 0.03 g-COD-cell/g-COD-substrate, suggesting that oxygen diffusion from the cathode possibly supported the microbial growth. In a comparative test, the biomass yield under aerobic environment was 0.46 ± 0.07 g-COD-cell/g-COD-substrate, which was about 3 times higher than the total biomass value in the MFC operation.

  4. Effect of dietary concentrate on rumen fermentation, digestibility, and nitrogen losses in dairy cows.

    PubMed

    Agle, M; Hristov, A N; Zaman, S; Schneider, C; Ndegwa, P M; Vaddella, V K

    2010-09-01

    The objective of this experiment was to investigate the effect of level of dietary concentrate on rumen fermentation, digestibility, and N losses in lactating dairy cows. The experiment was a replicated 3x3 Latin square design with 6 cows and 16-d adaptation periods. Ruminal contents were exchanged between cows at the beginning of each adaptation period. Data for 2 of the diets tested in this experiment are presented here. The diets contained (dry matter basis): 52% (LowC; control) and 72% (HighC) concentrate feeds. Crude protein contents of the diets were 16.5 and 16.4%, respectively. The HighC diet decreased ruminal pH and ammonia concentration and increased propionate concentration compared with LowC. Acetate:propionate ratio was greater for LowC than for HighC. Rumen methane production and microbial protein synthesis were unaffected by diet. Dry matter intake was similar among diets, but milk yield was increased by HighC compared with LowC (36.0 and 33.2 kg/d, respectively). Milk fat percentage and yield and total-tract apparent NDF digestibility were decreased by HighC compared with LowC. More ruminal ammonia N was transferred into milk protein with HighC than with LowC. Urinary N excretion, plasma urea N, and milk urea N concentration were not affected by diet. The ammonia emitting potential of manure was similar between LowC and HighC diets. Increased concentrate proportion in the diet of dairy cows resulted in reduced ruminal ammonia concentration and enhanced ammonia utilization for milk protein synthesis. These effects, however, did not result in reduced urinary N losses and only marginally improved milk N efficiency. Increasing dietary concentrate was not a successful strategy to mitigate enteric methane production and ammonia emissions from manure.

  5. Interaction of molasses and monensin in alfalfa hay- or corn silage-based diets on rumen fermentation, total tract digestibility, and milk production by Holstein cows.

    PubMed

    Oelker, E R; Reveneau, C; Firkins, J L

    2009-01-01

    Sugar supplementation can stimulate rumen microbial growth and possibly fiber digestibility; however, excess ruminal carbohydrate availability relative to rumen-degradable protein (RDP) can promote energy spilling by microbes, decrease rumen pH, or depress fiber digestibility. Both RDP supply and rumen pH might be altered by forage source and monensin. Therefore, the objective of this study was to evaluate interactions of a sugar source (molasses) with monensin and 2 forage sources on rumen fermentation, total tract digestibility, and production and fatty acid composition of milk. Seven ruminally cannulated lactating Holstein cows were used in a 5 x 7 incomplete Latin square design with five 28-d periods. Four corn silage diets consisted of 1) control (C), 2) 2.6% molasses (M), 3) 2.6% molasses plus 0.45% urea (MU), or 4) 2.6% molasses plus 0.45% urea plus monensin sodium (Rumensin, at the intermediate dosage from the label, 16 g/909 kg of dry matter; MUR). Three chopped alfalfa hay diets consisted of 1) control (C), 2) 2.6% molasses (M), or 3) 2.6% molasses plus Rumensin (MR). Urea was added to corn silage diets to provide RDP comparable to alfalfa hay diets with no urea. Corn silage C and M diets were balanced to have 16.2% crude protein; and the remaining diets, 17.2% crude protein. Dry matter intake was not affected by treatment, but there was a trend for lower milk production in alfalfa hay diets compared with corn silage diets. Despite increased total volatile fatty acid and acetate concentrations in the rumen, total tract organic matter digestibility was lower for alfalfa hay-fed cows. Rumensin did not affect volatile fatty acid concentrations but decreased milk fat from 3.22 to 2.72% in corn silage diets but less in alfalfa hay diets. Medium-chain milk fatty acids (% of total fat) were lower for alfalfa hay compared with corn silage diets, and short-chain milk fatty acids tended to decrease when Rumensin was added. In whole rumen contents, concentrations of

  6. Feeding, evaluating, and controlling rumen function

    USDA-ARS?s Scientific Manuscript database

    Optimal rumen function is pivotal to health, production, and profit in dairy systems. Achieving that function requires an understanding of feeds and systems of nutritional evaluation. A key influence on optimal rumen function includes dry matter intake. The function of feeds in the rumen depends on ...

  7. Effect of Origanum vulgare L. leaves on rumen fermentation, production, and milk fatty acid composition in lactating dairy cows.

    PubMed

    Hristov, A N; Lee, C; Cassidy, T; Heyler, K; Tekippe, J A; Varga, G A; Corl, B; Brandt, R C

    2013-02-01

    This experiment investigated the effects of dietary supplementation of Origanum vulgare L. leaf material (OR) on rumen fermentation, production, and milk fatty acid composition in dairy cows. The experimental design was a replicated 4 × 4 Latin square with 8 rumen-cannulated Holstein cows and 20-d experimental periods. Treatments were control (no OR supplementation), 250 g/cow per day OR (LOR), 500 g/d OR (MOR), and 750 g/d OR (HOR). Oregano supplementation had no effect on rumen pH, volatile fatty acid concentrations, and estimated microbial protein synthesis, but decreased ammonia concentration and linearly decreased methane production per unit of dry matter intake (DMI) compared with the unsupplemented control: 18.2, 16.5, 11.7, and 13.6g of methane/kg of DMI, respectively. Proportions of rumen bacterial, methanogen, and fungal populations were not affected by treatment. Treatment had no effect on total-tract apparent digestibility of dietary nutrients, except neutral detergent fiber digestibility was slightly decreased by all OR treatments compared with the control. Urinary N losses and manure odor were not affected by OR, except the proportion of urinary urea N in the total excreted urine N tended to be decreased compared with the control. Oregano linearly decreased DMI (28.3, 28.3, 27.5, and 26.7 kg/d for control, LOR, MOR, and HOR, respectively). Milk yield was not affected by treatment: 43.4, 45.2, 44.1, and 43.4 kg/d, respectively. Feed efficiency was linearly increased with OR supplementation and was greater than the control (1.46, 1.59, 1.60, and 1.63 kg/kg, respectively). Milk composition was unaffected by OR, except milk urea-N concentration was decreased. Milk fatty acid composition was not affected by treatment. In this short-term study, OR fed at 250 to 750 g/d decreased rumen methane production in dairy cows within 8h after feeding, but the effect over a 24-h feeding cycle has not been determined. Supplementation of the diet with OR linearly

  8. Microbial protein: future sustainable food supply route with low environmental footprint.

    PubMed

    Matassa, Silvio; Boon, Nico; Pikaar, Ilje; Verstraete, Willy

    2016-09-01

    Microbial biotechnology has a long history of producing feeds and foods. The key feature of today's market economy is that protein production by conventional agriculture based food supply chains is becoming a major issue in terms of global environmental pollution such as diffuse nutrient and greenhouse gas emissions, land use and water footprint. Time has come to re-assess the current potentials of producing protein-rich feed or food additives in the form of algae, yeasts, fungi and plain bacterial cellular biomass, producible with a lower environmental footprint compared with other plant or animal-based alternatives. A major driver is the need to no longer disintegrate but rather upgrade a variety of low-value organic and inorganic side streams in our current non-cyclic economy. In this context, microbial bioconversions of such valuable matters to nutritive microbial cells and cell components are a powerful asset. The worldwide market of animal protein is of the order of several hundred million tons per year, that of plant protein several billion tons of protein per year; hence, the expansion of the production of microbial protein does not pose disruptive challenges towards the process of the latter. Besides protein as nutritive compounds, also other cellular components such as lipids (single cell oil), polyhydroxybuthyrate, exopolymeric saccharides, carotenoids, ectorines, (pro)vitamins and essential amino acids can be of value for the growing domain of novel nutrition. In order for microbial protein as feed or food to become a major and sustainable alternative, addressing the challenges of creating awareness and achieving public and broader regulatory acceptance are real and need to be addressed with care and expedience.

  9. Functional Metagenomics Unveils a Multifunctional Glycosyl Hydrolase from the Family 43 Catalysing the Breakdown of Plant Polymers in the Calf Rumen

    PubMed Central

    Vieites, José María; López-Cortés, Nieves; Marín-Navarro, Julia; Nechitaylo, Taras Y.; Guazzaroni, María-Eugenia; Polaina, Julio; Waliczek, Agnes; Chernikova, Tatyana N.; Reva, Oleg N.; Golyshina, Olga V.; Golyshin, Peter N.

    2012-01-01

    Microbial communities from cow rumen are known for their ability to degrade diverse plant polymers at high rates. In this work, we identified 15 hydrolases through an activity-centred metagenome analysis of a fibre-adherent microbial community from dairy cow rumen. Among them, 7 glycosyl hydrolases (GHs) and 1 feruloyl esterase were successfully cloned, expressed, purified and characterised. The most striking result was a protein of GH family 43 (GHF43), hereinafter designated as R_09-02, which had characteristics very distinct from the other proteins in this family with mono-functional β-xylosidase, α-xylanase, α-L-arabinase and α-L-arabinofuranosidase activities. R_09-02 is the first multifunctional enzyme to exhibit β-1,4 xylosidase, α-1,5 arabinofur(pyr)anosidase, β-1,4 lactase, α-1,6 raffinase, α-1,6 stachyase, β-galactosidase and α-1,4 glucosidase activities. The R_09-02 protein appears to originate from the chromosome of a member of Clostridia, a class of phylum Firmicutes, members of which are highly abundant in ruminal environment. The evolution of R_09-02 is suggested to be driven from the xylose- and arabinose-specific activities, typical for GHF43 members, toward a broader specificity to the glucose- and galactose-containing components of lignocellulose. The apparent capability of enzymes from the GHF43 family to utilise xylose-, arabinose-, glucose- and galactose-containing oligosaccharides has thus far been neglected by, or could not be predicted from, genome and metagenome sequencing data analyses. Taking into account the abundance of GHF43-encoding gene sequences in the rumen (up to 7% of all GH-genes) and the multifunctional phenotype herein described, our findings suggest that the ecological role of this GH family in the digestion of ligno-cellulosic matter should be significantly reconsidered. PMID:22761666

  10. Metagenomic Analysis of the Rumen Microbiome of Steers with Wheat-Induced Frothy Bloat

    PubMed Central

    Pitta, D. W.; Pinchak, W. E.; Indugu, N.; Vecchiarelli, B.; Sinha, R.; Fulford, J. D.

    2016-01-01

    Frothy bloat is a serious metabolic disorder that affects stocker cattle grazing hard red winter wheat forage in the Southern Great Plains causing reduced performance, morbidity, and mortality. We hypothesize that a microbial dysbiosis develops in the rumen microbiome of stocker cattle when grazing on high quality winter wheat pasture that predisposes them to frothy bloat risk. In this study, rumen contents were harvested from six cannulated steers grazing hard red winter wheat (three with bloat score “2” and three with bloat score “0”), extracted for genomic DNA and subjected to 16S rDNA and shotgun sequencing on 454/Roche platform. Approximately 1.5 million reads were sequenced, assembled and assigned for phylogenetic and functional annotations. Bacteria predominated up to 84% of the sequences while archaea contributed to nearly 5% of the sequences. The abundance of archaea was higher in bloated animals (P < 0.05) and dominated by Methanobrevibacter. Predominant bacterial phyla were Firmicutes (65%), Actinobacteria (13%), Bacteroidetes (10%), and Proteobacteria (6%) across all samples. Genera from Firmicutes such as Clostridium, Eubacterium, and Butyrivibrio increased (P < 0.05) while Prevotella from Bacteroidetes decreased in bloated samples. Co-occurrence analysis revealed syntrophic associations between bacteria and archaea in non-bloated samples, however; such interactions faded in bloated samples. Functional annotations of assembled reads to Subsystems database revealed the abundance of several metabolic pathways, with carbohydrate and protein metabolism well represented. Assignment of contigs to CaZy database revealed a greater diversity of Glycosyl Hydrolases dominated by oligosaccharide breaking enzymes (>70%) in non-bloated samples. However, the abundance and diversity of CaZymes were greatly reduced in bloated samples indicating the disruption of carbohydrate metabolism. We conclude that mild to moderate frothy bloat results from tradeoffs both

  11. Microbial Protein-Protein Interactions (MiPPI) Data from the Genomics: GTL Center for Molecular and Cellular Systems (CMCS)

    DOE Data Explorer

    The Genomic Science Center for Molecular and Cellular Systems (CMCS), established in 2002, seeks to identify and characterize the complete set of protein complexes within a cell to provide a mechanistic basis for the understanding of biochemical functions. The CMCS is anchored at ORNL and PNNL. CMCS initially focused on the identification and characterization of protein complexes in two microbial systems,Rhodopseudomonas palustris (R. palustris) and Shewanella oneidensis (S. oneidensis). These two organisms have also been the focus of major DOE Genomic Science/Microbial Cell Program (MCP) projects. To develop an approach for identifying the diverse types of complexes present in microbial organisms, CMCS incorporates a number of molecular biology, microbiology, analytical and computational tools in an integrated pipeline.

  12. Next generation sequencing to define prokaryotic and fungal diversity in the bovine rumen.

    PubMed

    Fouts, Derrick E; Szpakowski, Sebastian; Purushe, Janaki; Torralba, Manolito; Waterman, Richard C; MacNeil, Michael D; Alexander, Leeson J; Nelson, Karen E

    2012-01-01

    A combination of Sanger and 454 sequences of small subunit rRNA loci were used to interrogate microbial diversity in the bovine rumen of 12 cows consuming a forage diet. Observed bacterial species richness, based on the V1-V3 region of the 16S rRNA gene, was between 1,903 to 2,432 species-level operational taxonomic units (OTUs) when 5,520 reads were sampled per animal. Eighty percent of species-level OTUs were dominated by members of the order Clostridiales, Bacteroidales, Erysipelotrichales and unclassified TM7. Abundance of Prevotella species varied widely among the 12 animals. Archaeal species richness, also based on 16S rRNA, was between 8 and 13 OTUs, representing 5 genera. The majority of archaeal OTUs (84%) found in this study were previously observed in public databases with only two new OTUs discovered. Observed rumen fungal species richness, based on the 18S rRNA gene, was between 21 and 40 OTUs with 98.4-99.9% of OTUs represented by more than one read, using Good's coverage. Examination of the fungal community identified numerous novel groups. Prevotella and Tannerella were overrepresented in the liquid fraction of the rumen while Butyrivibrio and Blautia were significantly overrepresented in the solid fraction of the rumen. No statistical difference was observed between the liquid and solid fractions in biodiversity of archaea and fungi. The survey of microbial communities and analysis of cross-domain correlations suggested there is a far greater extent of microbial diversity in the bovine rumen than previously appreciated, and that next generation sequencing technologies promise to reveal novel species, interactions and pathways that can be studied further in order to better understand how rumen microbial community structure and function affects ruminant feed efficiency, biofuel production, and environmental impact.

  13. Can perfluoroalkyl acids biodegrade in the rumen simulation technique (RUSITEC)?

    PubMed

    Kowalczyk, J; Riede, S; Schafft, H; Breves, G; Lahrssen-Wiederholt, M

    2015-01-01

    The behaviour of perfluoroalkyl acids (PFAAs) in tissues of ruminants has been shown to differ from that of monogastrics (J Agric Food Chem 61(12):2903-2912 doi:10.1021/jf304680j, 2013; J Agric Food Chem 62(28):6861-6870, 2014). This may be a consequence of the complex microbial ecosystem in the rumen. To evaluate this hypothesis, the recovery of PFAAs was studied using the rumen simulation technique as an indication for biodegradation in rumen. The PFAA-recovery from a microbial fermentation of feed containing PFAAs was compared to the same feed in the absence of ruminal microorganisms (MOs). Release of PFAAs from feed into fermentation fluid was found to be faster for perfluorobutane sulfonic acid (PFBS) than for perfluorooctane sulfonic acid (PFOS). Differences between perfluoroalkyl carboxylic acids (PFCAs) could not be observed. Proportions of PFAAs recovered in the fermentation fluids decreased by increasing chain lengths for the perfluoroalkyl sulfonic acids (PFSAs) (31 % PFBS, 28 % perfluorohexane sulfonic acid [PFHxS], 20 % perfluoroheptane sulfonic acid [PFHpS], 11 % PFOS) and PFCAs (33 % perfluorohexane carboxylic acid [PFHxA], 32 % perfluoroheptane carboxylic acid [PFHpA], 24 % perfluorooctanoic acid [PFOA]). In contrast, levels in feed increased with increasing chain length for both PFSAs and PFCAs. The attachment of MOs to feed particles was assumed to account for higher PFAA levels in fermented feeds and for lower levels in the fermentation fluids. Total recovery of PFAAs was significantly lower in presence of ruminal MOs compared to experimental procedure under sterile conditions. Although, there are optimal reductive conditions for MOs in rumen, our results do not univocally indicate whether PFAAs were degraded by ruminal fermentation.

  14. Supplementation with non-fibrous carbohydrates reduced fiber digestibility and did not improve microbial protein synthesis in sheep fed fresh forage of two nutritive values.

    PubMed

    Tebot, I; Cajarville, C; Repetto, J L; Cirio, A

    2012-04-01

    To determine whether non-fibrous carbohydrate (NFC) supplementation improves fiber digestibility and microbial protein synthesis, 18 Corriedale ewes with a fixed intake level (40 g dry matter (DM)/kg BW0.75) were assigned to three (n = 6) diets: F = 100% fresh temperate forage, FG = 70% forage + 30% barley grain and FGM = 70% forage + 15% barley grain + 15% molasses-based product (MBP, Kalori 3000). Two experimental periods were carried out, with late (P1) and early (P2) vegetative stage forage. For P2, ewes were fitted with ruminal catheters. Forage was distributed at 0900 h, 1300 h, 1800 h and 2300 h, and supplement added at 0900 h and 1800 h meals. Digestibility of the different components of the diets, retained N and rumen microbial protein synthesis were determined. At the end of P2, ruminal pH and N-NH3 concentration were determined hourly for 24 h. Supplementation increased digestibility of DM (P < 0.001) and organic matter (OM; P < 0.001) and reduced NDF digestibility (P = 0.043) in both periods, with greater values in P2 (P = 0.008) for the three diets. Daily mean ruminal pH differed (P < 0.05) among treatments: 6.33 (F), 6.15 (FG) and 6.51 (FGM). The high pH in FGM was attributed to Ca(OH)2 in MBP. Therefore, the decreased fiber digestibility in supplemented diets could not be attributed to pH changes. The mean ruminal concentration of N-NH3 was 18.0 mg/dl, without differences among treatments or sampling hours. Microbial protein synthesis was greater in P2 (8.0 g/day) than in P1 (6.1 g/day; P = 0.006), but treatments did not enhance this parameter. The efficiency of protein synthesis tended to be lower in supplemented groups (16.4, 13.9 and 13.4 in P1, and 20.8, 16.7 and 16.2 g N/kg digestible OM ingested in P2, for F, FG and FGM, respectively; P = 0.07) without differences between supplements. The same tendency was observed for retained N: 2.55, 1.38 and 1.98 in P1, and 2.28, 1.23 and 1.10 g/day in P2, for F, FG and FGM, respectively; P = 0.05). The

  15. Effect of dietary starch level and high rumen-undegradable protein on endocrine-metabolic status, milk yield, and milk composition in dairy cows during early and late lactation.

    PubMed

    Piccioli-Cappelli, F; Loor, J J; Seal, C J; Minuti, A; Trevisi, E

    2014-12-01

    Diet composition defines the amount and type of nutrients absorbed by dairy cows. Endocrine-metabolic interactions can influence these parameters, and so nutrient availability for the mammary gland can significantly vary and affect milk yield and its composition. Six dairy cows in early and then late lactation received, for 28 d in a changeover design, 2 diets designed to provide, within the same stage of lactation, similar amounts of rumen fermentable material but either high starch plus sugar (HS) content or low starch plus sugar content (LS). All diets had similar dietary crude protein and calculated supply of essential amino acids. Dry matter intake within each stage of lactation was similar between groups. Milk yield was similar between groups in early lactation, whereas a higher milk yield was observed in late lactation when feeding HS. At the metabolic level, the main difference observed between the diets in both stages of lactation was lower blood glucose in cows fed LS. The lower glucose availability during consumption of LS caused substantial modifications in the circulating and postprandial pattern of metabolic hormones. Feeding LS versus HS resulted in an increase in the ratio of bovine somatotropin to insulin. This increased mobilization of lipid reserves resulted in higher blood concentrations of nonesterified fatty acids and β-hydroxybutyrate, which contributed to the higher milk fat content in both stages of lactation in the LS group. This greater recourse to body fat stores was confirmed by the greater loss of body weight during early lactation and the slower recovery of body weight in late lactation in cows fed LS. The lower insulin to glucagon ratio observed in cows fed LS in early and late lactation likely caused an increase in hepatic uptake and catabolism of amino acids, as confirmed by the higher blood urea concentrations. Despite the higher catabolism of amino acids in LS in early lactation, similar milk protein output was observed for both

  16. Structure and Function of Microbial Metal-Reduction Proteins

    SciTech Connect

    Xu, Ying; Crawford, Oakly H.; Xu, Dong; Larimer, Frank W.; Uberbacher, Edward C.; Zhou, Jizhong

    2009-09-02

    In this project, we proposed (i) identification of metal-reduction genes, (ii) development of new threading techniques and (iii) fold recognition and structure prediction of metal-reduction proteins. However, due to the reduction of the budget, we revised our plan to focus on two specific aims of (i) developing a new threading-based protein structure prediction method, and (ii) developing an expert system for protein structure prediction.

  17. Supplementation of Flemingia macrophylla and cassava foliage as a rumen enhancer on fermentation efficiency and estimated methane production in dairy steers.

    PubMed

    Phesatcha, Burarat; Wanapat, Metha; Phesatcha, Kampanat; Ampapon, Thiwakorn; Kang, Sungchhang

    2016-10-01

    Four rumen-fistulated dairy steers, 3 years old with 180 ± 15 kg body weight (BW), were randomly assigned according to a 4 × 4 Latin square design to investigate on the effect of Flemingia macrophylla hay meal (FMH) and cassava hay meal (CH) supplementation on rumen fermentation efficiency and estimated methane production. The treatments were as follows: T1 = non-supplement, T2 = CH supplementation at 150 g/head/day, T3 = FMH supplementation at 150 g/head/day, and T4 = CH + FMH supplementation at 75 and 75 g/head/day. All steers were fed rice straw ad libitum and concentrate was offered at 0.5 % of BW. Results revealed that supplementation of CH and/or FMH did not affect on feed intake (P > 0.05) while digestibility of crude protein and neutral detergent fiber were increased especially in steers receiving FMH and CH+FMH (P < 0.05). Ruminal pH, temperature, and blood urea nitrogen were similar among treatments while ammonia nitrogen was increased in all supplemented groups (P < 0.05). Furthermore, propionic acid (C3) was increased while acetic acid (C2), C2:C3 ratio, and estimated methane production were decreased by dietary treatments. Protozoa and fungi population were not affected by dietary supplement while viable bacteria count increased in steers receiving FMH. Supplementation of FMH and/or FMH+CH increased microbial crude protein and efficiency of microbial nitrogen supply. This study concluded FMH (150 g/head/day) and/or CH+FMH (75 and 75 g/head/day) supplementation could be used as a rumen enhancer for increasing nutrient digestibility, rumen fermentation efficiency, and microbial protein synthesis while decreasing estimated methane production without adverse effect on voluntary feed intake of dairy steers fed rice straw.

  18. Next generation sequencing to define prokaryotic and fungal diversity in the bovine rumen

    USDA-ARS?s Scientific Manuscript database

    A combination of Sanger and 454 sequences of small subunit rRNA loci were used to interrogate the microbial diversity in the bovine rumen of 14 pasture-fed animals. The observed bacterial species richness, based on the V1-V3 region of the 15S rRNA gene, was between 1902 to 2596 species-level operati...

  19. Transcriptome differences in the rumen of beef steers with variation in feed intake and gain

    USDA-ARS?s Scientific Manuscript database

    Feed efficiency is an economically important trait in beef production. The rumen wall interacts with feed, microbial populations and volatile fatty acids important to ruminant nutrition indicating it may play a critical role in the beef steer’s ability to utilize feedstuffs efficiently. To identif...

  20. Determining the amount of rumen-protected methionine supplement that corresponds to the optimal levels of methionine in metabolizable protein for maximizing milk protein production and profit on dairy farms.

    PubMed

    Cho, J; Overton, T R; Schwab, C G; Tauer, L W

    2007-10-01

    The profitability of feeding rumen-protected Met (RPMet) sources to produce milk protein was estimated using a 2-step procedure: First, the effect of Met in metabolizable protein (MP) on milk protein production was estimated by using a quadratic Box-Cox functional form. Then, using these estimation results, the amounts of RPMet supplement that corresponded to the optimal levels of Met in MP for maximizing milk protein production and profit on dairy farms were determined. The data used in this study were modified from data used to determine the optimal level of Met in MP for lactating cows in the Nutrient Requirements of Dairy Cattle (NRC, 2001). The data used in this study differ from that in the NRC (2001) data in 2 ways. First, because dairy feed generally contains 1.80 to 1.90% Met in MP, this study adjusts the reference production value (RPV) from 2.06 to 1.80 or 1.90%. Consequently, the milk protein production response is also modified to an RPV of 1.80 or 1.90% Met in MP. Second, because this study is especially interested in how much additional Met, beyond the 1.80 or 1.90% already contained in the basal diet, is required to maximize farm profits, the data used are limited to concentrations of Met in MP above 1.80 or 1.90%. This allowed us to calculate any additional cost to farmers based solely on the price of an RPMet supplement and eliminated the need to estimate the dollar value of each gram of Met already contained in the basal diet. Results indicated that the optimal level of Met in MP for maximizing milk protein production was 2.40 and 2.42%, where the RPV was 1.80 and 1.90%, respectively. These optimal levels were almost identical to the recommended level of Met in MP of 2.40% in the NRC (2001). The amounts of RPMet required to increase the percentage of Met in MP from each RPV to 2.40 and 2.42% were 21.6 and 18.5 g/d, respectively. On the other hand, the optimal levels of Met in MP for maximizing profit were 2.32 and 2.34%, respectively. The amounts

  1. Effects and mode of action of chitosan and ivy fruit saponins on the microbiome, fermentation and methanogenesis in the rumen simulation technique.

    PubMed

    Belanche, Alejandro; Pinloche, Eric; Preskett, David; Newbold, C Jamie

    2016-01-01

    This study investigates the effects of supplementing a control diet (CON) with chitosan (CHI) or ivy fruit saponins (IVY) as natural feed additives. Both additives had similar abilities to decrease rumen methanogenesis (-42% and -40%, respectively) using different mechanisms: due to its antimicrobial and nutritional properties CHI promoted a shift in the fermentation pattern towards propionate production which explained about two thirds of the decrease in methanogenesis. This shift was achieved by a simplification of the structure in the bacterial community and a substitution of fibrolytic (Firmicutes and Fibrobacteres) by amylolytic bacteria (Bacteroidetes and Proteobacteria) which led to greater amylase activity, lactate and microbial protein yield with no detrimental effect on feed digestibility. Contrarily, IVY had negligible nutritional properties promoting minor changes in the fermentation pattern and on the bacterial community. Instead, IVY modified the structure of the methanogen community and decreased its diversity. This specific antimicrobial effect of IVY against methanogens was considered its main antimethanogenic mechanism. IVY had however a negative impact on microbial protein synthesis. Therefore, CHI and IVY should be further investigated in vivo to determine the optimum doses which maintain low methanogenesis but prevent negative effects on the rumen fermentation and animal metabolism.

  2. Monensin and Nisin Affect Rumen Fermentation and Microbiota Differently In Vitro

    PubMed Central

    Shen, Junshi; Liu, Zhuang; Yu, Zhongtang; Zhu, Weiyun

    2017-01-01

    Nisin, a bacteriocin, is a potential alternative to antibiotics to modulate rumen fermentation. However, little is known about its impacts on rumen microbes. This study evaluated the effects of nisin (1 and 5 μM) on in vitro rumen fermentation characteristics, microbiota, and select groups of rumen microbes in comparison with monensin (5 μM), one of the most commonly used ionophores in ruminants. Nisin had greater effects than monensin in inhibiting methane production and decreasing acetate/propionate ratio. Unlike monensin, nisin had no adverse effect on dry matter digestibility. Real-time PCR analysis showed that both monensin and nisin reduced the populations of total bacteria, fungi, and methanogens, while the population of protozoa was reduced only by monensin. Principal component analysis of bacterial 16S rRNA gene amplicons showed a clear separation between the microbiota shaped by monensin and by nisin. Comparative analysis also revealed a significant difference in relative abundance of some bacteria in different taxa between monensin and nisin. The different effects of monensin and nisin on microbial populations and bacterial communities are probably responsible for the discrepancy in their effects on rumen fermentation. Nisin may have advantages over monensin in modulating ruminal microbial ecology and reducing ruminant methane production without adversely affecting feed digestion, and thus it may be used as a potential alternative to monensin fed to ruminants. PMID:28670304

  3. The effect of dietary Chlorella vulgaris supplementation on micro-organism community, enzyme activities and fatty acid profile in the rumen liquid of goats.

    PubMed

    Tsiplakou, E; Abdullah, M A M; Skliros, D; Chatzikonstantinou, M; Flemetakis, E; Labrou, N; Zervas, G

    2017-04-01

    Microalgae might be considered as an alternative source of fat and/or protein for ruminant's diets. However, changes in populations of ruminal micro-organisms associated with biohydrogenation process, methane and ammonia production in response to microalgae dietary supplementation have not been well characterized. Thus, 16 cross-bred goats were divided into two groups. Each goat of both groups was fed individually with alfalfa hay and concentrates separately. The concentrates of the control group had no microalgae while those of the treated group were supplemented with 10 g lyophilized Chlorella vulgaris/kg concentrate (chlor). On the 30th experimental day, samples of rumen fluid were collected for microbial DNA extraction, fatty acid profile and enzyme activity analyses. The results showed that the chlor diet compared with the control increased significantly the populations of Methanosphaera stadtmanae, Methanobrevibacter ruminantium and Methanogens bacteria and protozoa in the rumen of goats. A significant reduction in the cellulase activity and in the abundance of Ruminococcus albus, and a significant increase in the protease activity and in the abundance of Clostridium sticklandii in the rumen liquid of goats fed with the chlor diet, compared with the control, were found. Chlorella vulgaris supplementation promoted the formation of trans C18:1 , trans-11 C18:1 and monounsaturated fatty acids (MUFA), while the proportions of C18:0 and long-chain fatty acids (LCFA) reduced significantly in the rumen liquid of goats. This shift in ruminal biohydrogenation pathway was accompanied by a significant increase in Butyrivibrio fibrisolvens trans C18:1 -producing bacteria. In conclusion, the supplementation of diets with microalgae needs further investigation because it enhances the populations of methane-producing bacteria and protozoa.

  4. Identification of a Class of Protein ADP-Ribosylating Sirtuins in Microbial Pathogens

    PubMed Central

    Rack, Johannes Gregor Matthias; Morra, Rosa; Barkauskaite, Eva; Kraehenbuehl, Rolf; Ariza, Antonio; Qu, Yue; Ortmayer, Mary; Leidecker, Orsolya; Cameron, David R.; Matic, Ivan; Peleg, Anton Y.; Leys, David; Traven, Ana; Ahel, Ivan

    2015-01-01

    Summary Sirtuins are an ancient family of NAD+-dependent deacylases connected with the regulation of fundamental cellular processes including metabolic homeostasis and genome integrity. We show the existence of a hitherto unrecognized class of sirtuins, found predominantly in microbial pathogens. In contrast to earlier described classes, these sirtuins exhibit robust protein ADP-ribosylation activity. In our model organisms, Staphylococcus aureus and Streptococcus pyogenes, the activity is dependent on prior lipoylation of the target protein and can be reversed by a sirtuin-associated macrodomain protein. Together, our data describe a sirtuin-dependent reversible protein ADP-ribosylation system and establish a crosstalk between lipoylation and mono-ADP-ribosylation. We propose that these posttranslational modifications modulate microbial virulence by regulating the response to host-derived reactive oxygen species. PMID:26166706

  5. Microbial drug efflux proteins of the major facilitator superfamily.

    PubMed

    Saidijam, Massoud; Benedetti, Giulia; Ren, Qinghu; Xu, Zhiqiang; Hoyle, Christopher J; Palmer, Sarah L; Ward, Alison; Bettaney, Kim E; Szakonyi, Gerda; Meuller, Johan; Morrison, Scott; Pos, Martin K; Butaye, Patrick; Walravens, Karl; Langton, Kate; Herbert, Richard B; Skurray, Ronald A; Paulsen, Ian T; O'reilly, John; Rutherford, Nicholas G; Brown, Melissa H; Bill, Roslyn M; Henderson, Peter J F

    2006-07-01

    Drug efflux proteins are widespread amongst microorganisms, including pathogens. They can contribute to both natural insensitivity to antibiotics and to emerging antibiotic resistance and so are potential targets for the development of new antibacterial drugs. The design of such drugs would be greatly facilitated by knowledge of the structures of these transport proteins, which are poorly understood, because of the difficulties of obtaining crystals of quality. We describe a structural genomics approach for the amplified expression, purification and characterisation of prokaryotic drug efflux proteins of the 'Major Facilitator Superfamily' (MFS) of transport proteins from Helicobacter pylori, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, Bacillus subtilis, Brucella melitensis, Campylobacter jejuni, Neisseria meningitides and Streptomyces coelicolor. The H. pylori putative drug resistance protein, HP1092, and the S. aureus QacA proteins are used as detailed examples. This strategy is an important step towards reproducible production of transport proteins for the screening of drug binding and for optimisation of crystallisation conditions to enable subsequent structure determination.

  6. Utilization of digital differential display to identify differentially expressed genes related to rumen development.

    PubMed

    Kato, Daichi; Suzuki, Yutaka; Haga, Satoshi; So, KyoungHa; Yamauchi, Eri; Nakano, Miwa; Ishizaki, Hiroshi; Choi, Kichoon; Katoh, Kazuo; Roh, Sang-Gun

    2016-04-01

    This study aimed to identify the genes associated with the development of the rumen epithelium by screening for candidate genes by digital differential display (DDD) in silico. Using DDD in NCBI's UniGene database, expressed sequence tag (EST)-based gene expression profiles were analyzed in rumen, reticulum, omasum, abomasum and other tissues in cattle. One hundred and ten candidate genes with high expression in the rumen were derived from a library of all tissues. The expression levels of 11 genes in all candidate genes were analyzed in the rumen, reticulum, omasum and abomasum of nine Japanese Black male calves (5-week-old pre-weaning: n = 3; 15-week-old weaned calves: n = 6). Among the 11 genes, only 3-hydroxy-3-methylglutaryl-CoA synthase 2 (HMGCS2), aldo-keto reductase family 1, member C1-like (AKR1C1), and fatty acid binding protein 3 (FABP3) showed significant changes in the levels of gene expression in the rumen between the pre- and post-weaning of calves. These results indicate that DDD analysis in silico can be useful for screening candidate genes related to rumen development, and that the changes in expression levels of three genes in the rumen may have been caused by weaning, aging or both.

  7. Microbial response to single-cell protein production and brewery wastewater treatment

    PubMed Central

    Lee, Jackson Z; Logan, Andrew; Terry, Seth; Spear, John R

    2015-01-01

    As global fisheries decline, microbial single-cell protein (SCP) produced from brewery process water has been highlighted as a potential source of protein for sustainable animal feed. However, biotechnological investigation of SCP is difficult because of the natural variation and complexity of microbial ecology in wastewater bioreactors. In this study, we investigate microbial response across a full-scale brewery wastewater treatment plant and a parallel pilot bioreactor modified to produce an SCP product. A pyrosequencing survey of the brewery treatment plant showed that each unit process selected for a unique microbial community. Notably, flow equalization basins were dominated by Prevotella, methanogenesis effluent had the highest levels of diversity, and clarifier wet-well samples were sources of sequences for the candidate bacterial phyla of TM7 and BD1-5. Next, the microbial response of a pilot bioreactor producing SCP was tracked over 1 year, showing that two different production trials produced two different communities originating from the same starting influent. However, SCP production resulted generally in enrichment of several clades of rhizospheric diazotrophs of Alphaproteobacteria and Betaproteobacteria in the bioreactor and even more so in the final product. These diazotrophs are potentially useful as the basis of a SCP product for commercial feed production. PMID:24837420

  8. A perfusion-capable microfluidic bioreactor for assessing microbial heterologous protein production

    PubMed Central

    Mozdzierz, Nicholas J.; Love, Kerry R.; Lee, Kevin S.; Lee, Harry L. T.; Shah, Kartik A.; Ram, Rajeev J.

    2015-01-01

    We present an integrated microfluidic bioreactor for fully continuous perfusion cultivation of suspended microbial cell cultures. This system allowed continuous and stable heterologous protein expression by sustaining the cultivation of Pichia pastoris over 11 days. This technical capability also allowed testing the impact of perfusion conditions on protein expression. This advance should enable small-scale models for process optimization in continuous biomanufacturing. PMID:26055071

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

    PubMed

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

    2016-12-01

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

  10. Downregulation of Cellular Protective Factors of Rumen Epithelium in Goats Fed High Energy Diet

    PubMed Central

    Hollmann, Manfred; Miller, Ingrid; Hummel, Karin; Sabitzer, Sonja; Metzler-Zebeli, Barbara U.; Razzazi-Fazeli, Ebrahim; Zebeli, Qendrim

    2013-01-01

    Energy-rich diets can challenge metabolic and protective functions of the rumen epithelial cells, but the underlying factors are unclear. This study sought to evaluate proteomic changes of the rumen epithelium in goats fed a low, medium, or high energy diet. Expression of protein changes were compared by two-dimensional differential gel electrophoresis followed by protein identification with matrix assisted laser desorption ionisation tandem time-of-flight mass spectrometry. Of about 2,000 spots commonly detected in all gels, 64 spots were significantly regulated, which were traced back to 24 unique proteins. Interestingly, the expression profiles of several chaperone proteins with important cellular protective functions such as heat shock cognate 71 kDa protein, peroxiredoxin-6, serpin H1, protein disulfide-isomerase, and selenium-binding protein were collectively downregulated in response to high dietary energy supply. Similar regulation patterns were obtained for some other proteins involved in transport or metabolic functions. In contrast, metabolic enzymes like retinal dehydrogenase 1 and ATP synthase subunit beta, mitochondrial precursor were upregulated in response to high energy diet. Lower expressions of chaperone proteins in the rumen epithelial cells in response to high energy supply may suggest that these cells were less protected against the potentially harmful rumen toxic compounds, which might have consequences for rumen and systemic health. Our findings also suggest that energy-rich diets and the resulting acidotic insult may render rumen epithelial cells more vulnerable to cellular damage by attenuating their cell defense system, hence facilitating the impairment of rumen barrier function, typically observed in energy-rich fed ruminants. PMID:24349094

  11. [Effects of zinc-finger proteins and artificial zinc-finger proteins on microbial metabolisms--a review].

    PubMed

    Liu, Zhuo; Zhang, Fei; Zhao, Xinqing; Bai, Fengwu

    2014-03-01

    Zinc-finger proteins have been widely studied due to their highly conserved structures and DNA-binding specificity of zinc-finger domains. However, researches on the zinc-finger proteins from microorganisms, especially those from prokaryotes, are still very limited. This review focuses on the latest progress on microbial zinc-finger proteins, especially those from prokaryotes and the application of artificial zinc-finger proteins in the breeding of robust strains. Artificial zinc-finger proteins with transcriptional activation or repression domain can regulate the global gene transcription of microbial cells to acquire improved phenotypes, such as stress tolerance to heat, ethanol, butanol, and osmotic pressure. Using the zinc-finger domain as DNA scaffold in the construction of enzymatic system can enhance the catalytic efficiency and subsequently the production of specific metabolites. Currently, zinc-finger domains used in the construction of artificial transcription factor are usually isolated from mammalian cells. In the near future, novel transcription factors can be designed for strain development based on the natural zinc-finger domains from different microbes, which may be used to regulate the global gene expression of microbial cells more efficiently.

  12. In vitro effects of sodium bicarbonate buffer on rumen fermentation, levels of lipopolysaccharide and biogenic amine, and composition of rumen microbiota.

    PubMed

    Mao, Shengyong; Huo, Wenjie; Liu, Junhua; Zhang, Ruiyang; Zhu, Weiyun

    2017-03-01

    Diets containing high levels of carbohydrates provoke a rapid decrease of rumen pH and high levels of biogenic amines and lipopolysaccharides (LPS), which severely impair the health and performance of ruminants. The goal of this study was to evaluate the effects of sodium bicarbonate (BC) buffer on rumen fermentation, levels of LPS and biogenic amine, and composition of rumen microbiota using in vitro rumen cultures. Sodium bicarbonate supplementation increased (P < 0.05) the final pH levels and concentrations of total volatile fatty acids and LPS, as well as the proportions of acetate, propionate, isobutyrate, isovalerate and valerate, and it decreased (P < 0.05) the proportion of butyrate and the levels of lactic acid, methylamine, tryptamine, tyramine, histamine and putrescine compared with the control. Pyrosequencing of the 16S rRNA gene showed that BC inclusion increased (P < 0.05) the bacterial diversity index compared with the control. Adding BC also decreased (P < 0.05) the relative abundance of Streptococcus and Butyrivibrio and increased (P < 0.05) the proportions of Ruminococcus, Succinivibrio and Prevotella. Sodium bicarbonate supplementation has beneficial effects in the reduction of bioamine levels and the increase in ruminal pH, and in modifying the microbial ecology of the rumen; however, it results in an accumulation of LPS under high-grain diet conditions. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  13. The effects of additives in napier grass silages on chemical composition, feed intake, nutrient digestibility and rumen fermentation.

    PubMed

    Bureenok, Smerjai; Yuangklang, Chalermpon; Vasupen, Kraisit; Schonewille, J Thomas; Kawamoto, Yasuhiro

    2012-09-01

    The effect of silage additives on ensiling characteristics and nutritive value of Napier grass (Pennisetum purpureum) silages was studied. Napier grass silages were made with no additive, fermented juice of epiphytic lactic acid bacteria (FJLB), molasses or cassava meal. The ensiling characteristics were determined by ensiling Napier grass silages in airtight plastic pouches for 2, 4, 7, 14, 21 and 45 d. The effect of Napier grass silages treated with these additives on voluntary feed intake, digestibility, rumen fermentation and microbial rumen fermentation was determined in 4 fistulated cows using 4×4 Latin square design. The pH value of the treated silages rapidly decreased, and reached to the lowest value within 7 d of the start of fermentation, as compared to the control. Lactic acid content of silages treated with FJLB was stable at 14 d of fermentation and constant until 45 d of ensiling. At 45 d of ensiling, neutral detergent fiber (NDF) and acid detergent fiber (ADF) of silage treated with cassava meal were significantly lower (p<0.05) than the others. In the feeding trial, the intake of silage increased (p<0.05) in the cow fed with the treated silage. Among the treatments, dry matter intake was the lowest in the silage treated with cassava meal. The organic matter, crude protein and NDF digestibility of the silage treated with molasses was higher than the silage without additive and the silage treated with FJLB. The rumen parameters: ruminal pH, ammonia-nitrogen (NH3-N), volatile fatty acid (VFA), blood urea nitrogen (BUN) and bacterial populations were not significantly different among the treatments. In conclusion, these studies confirmed that the applying of molasses improved fermentative quality, feed intake and digestibility of Napier grass.

  14. The Effects of Additives in Napier Grass Silages on Chemical Composition, Feed Intake, Nutrient Digestibility and Rumen Fermentation

    PubMed Central

    Bureenok, Smerjai; Yuangklang, Chalermpon; Vasupen, Kraisit; Schonewille, J. Thomas; Kawamoto, Yasuhiro

    2012-01-01

    The effect of silage additives on ensiling characteristics and nutritive value of Napier grass (Pennisetum purpureum) silages was studied. Napier grass silages were made with no additive, fermented juice of epiphytic lactic acid bacteria (FJLB), molasses or cassava meal. The ensiling characteristics were determined by ensiling Napier grass silages in airtight plastic pouches for 2, 4, 7, 14, 21 and 45 d. The effect of Napier grass silages treated with these additives on voluntary feed intake, digestibility, rumen fermentation and microbial rumen fermentation was determined in 4 fistulated cows using 4×4 Latin square design. The pH value of the treated silages rapidly decreased, and reached to the lowest value within 7 d of the start of fermentation, as compared to the control. Lactic acid content of silages treated with FJLB was stable at 14 d of fermentation and constant until 45 d of ensiling. At 45 d of ensiling, neutral detergent fiber (NDF) and acid detergent fiber (ADF) of silage treated with cassava meal were significantly lower (p<0.05) than the others. In the feeding trial, the intake of silage increased (p<0.05) in the cow fed with the treated silage. Among the treatments, dry matter intake was the lowest in the silage treated with cassava meal. The organic matter, crude protein and NDF digestibility of the silage treated with molasses was higher than the silage without additive and the silage treated with FJLB. The rumen parameters: ruminal pH, ammonia-nitrogen (NH3-N), volatile fatty acid (VFA), blood urea nitrogen (BUN) and bacterial populations were not significantly different among the treatments. In conclusion, these studies confirmed that the applying of molasses improved fermentative quality, feed intake and digestibility of Napier grass. PMID:25049687

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

    PubMed

    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.

  16. Human consumption of rumen flukes of cattle in India.

    PubMed

    Sarmah, P C; Laha, R; Bhattacharjee, K; Goswami, A; Raquib, M; Kakati, P

    2014-01-01

    The practice of eating rumen flukes of cattle by a section of people living in Meghalaya, a north eastern State of India, is reported in this communication. Economically backward, some rural people belonging to Khasi, Jaintia, Garo, and Karbi tribes of Christian and Nepali communities who eat beef are accustomed to consuming cooked flukes during breakfast, meals, and also along with rice beer or alcohol. Inspection of the rumens of cattle during slaughter indicated a prevalence of flukes belonging to Cotylophoron, Paramphistomum, Calicophoron, Gastrothylax, and Fischoederius genera in 74% cases, and their collection from rumen ranged approximately from 50 g to 600 g. Biochemical analysis of flukes found 12.60% total protein, 0.78% fat, and 0.87% ash on fresh weight basis. High prevalence of flukes, easy visualization in rumen, their bulk collection, presence of nutritive value, absence of any ill effect, and lack of imminent danger of transmissibility are believed to be the rationales influencing their consumption by people. It is suggested that dietary benefits obtained from flukes might contribute to the energy transfer and inclusion in the food web.

  17. Effect of rumen-protected methionine on feed intake, milk production, true milk protein concentration, and true milk protein yield, and the factors that influence these effects: a meta-analysis.

    PubMed

    Patton, R A

    2010-05-01

    A meta-analysis of published studies was used to investigate the effect of rumen-protected methionine (RPM) added to the diets of lactating dairy cattle on dry matter intake, milk production, true milk protein (TMP) production, and milk fat yield. Differences in responses between 2 commonly used RPM products, Mepron (Evonik Industries, Hanau, Germany) and Smartamine (Adisseo, Antony, France), were investigated as well as dietary and animal factors that could influence responses. Diets were coded with respect to the amino acid (AA) deficiency of the control diet as predicted by the AminoCow model (version 3.5.2, http://www.makemilknotmanure.com/aminocow.php; 0=no AA deficiency, 1=Met deficiency, 2=Met and Lys deficiency, 3=Met and Lys plus at least 1 other AA deficiency) to test the effect of AA deficiencies on RPM response. Thirty-five studies were identified, 17 studies evaluating Mepron, 18 studies evaluating Smartamine, and 1 study evaluating both. This permitted 75 dietary comparisons between control and RPM-added diets. Diets were entered into the AminoCow and the 2001 National Research Council models to compare predictions of Met, Lys, and metabolizable protein (MP) flow. Mean Met and Lys in diets where RPM was fed were estimated to be 2.35 and 6.33% of MP, respectively. Predictions of flows between models were similar. Overall, RPM addition to diets increased production of TMP, both as percentage (0.07%) and yield (27 g/d). Dry matter intake and milk fat percentage were slightly decreased, whereas milk production was slightly increased. Differences between products were detected for all production variables, with Mepron-fed cows producing less TMP percentage but greater milk production, resulting in twice as much TMP yield. Milk protein response to RPM was not related to predicted AA deficiency, calculated Met deficiency, or Met as a percentage of MP. Other dietary factors, including Lys flow (g/d), Lys as percentage of MP, neutral detergent fiber percentage

  18. Evaluation of isoquinoline alkaloid supplementation levels on ruminal fermentation, characteristics of digestion, and microbial protein synthesis in steers fed a high-energy diet.

    PubMed

    Aguilar-Hernández, J A; Urías-Estrada, J D; López-Soto, M A; Barreras, A; Plascencia, A; Montaño, M; González-Vizcarra, V M; Estrada-Angulo, A; Castro-Pérez, B I; Barajas, R; Rogge, H I; Zinn, R A

    2016-01-01

    Four Holstein steers with ruminal and duodenal cannulas were used in a 4 × 4 Latin square design to examine the effect of daily intake of 0, 2, 4 or 6 g/steer of standardized plant extract containing a mixture of quaternary benzophenanthridine alkaloids and protopine alkaloids (QBA+PA) on the characteristics of ruminal fermentation and characteristics of digestion. The basal diet consisted of a steam-flaked corn-based finishing diet that contained 62% corn and 12% sudangrass hay and the rest of diet was composed of mainly dried distillers grains, molasses, fat, and minerals. The source of QBA+PA used was Sangrovit-RS (Phytobiotics Futterzusatzstoffe GmbH, Eltville, Germany) and supplementation levels of 2, 4, and 6 g Sangrovit-RS∙steer∙d, which represented a net daily ingestion of approximately 6, 12, and 18 mg of QBA+PA compounds, respectively. Inclusion of QBA+PA linearly increased ( = 0.04) flow to the duodenum of nonammonia N and linearly decreased ( < 0.01) duodenal flows of ammonia N. Ruminal microbial efficiency (duodenal microbial N; g/kg OM fermented in the rumen) and protein efficiency (duodenal nonammonia N; g/g N intake) were increased ( < 0.05) as the level of QBA+PA increased. There were no effects of QBA+PA supplementation on ruminal, postruminal, and total tract digestion of OM, starch, and NDF, but postruminal and total tract digestion of N increased ( < 0.01) as the level of QBA+PA increased. Digestible energy of the diet tended to increase (linear affect, = 0.09) with QBA+PA supplementation. Ruminal pH and total VFA molar concentrations were not different between treatments. Ruminal NH-N concentration linearly decreased ( = 0.02) with QBA+PA supplementation. Ruminal molar proportion of acetate increased ( = 0.04) as the supplementation level of QBA+PA increased. It is concluded that QBA+PA supplementation enhances efficiency of N utilization in feedlot steers fed a steam-flaked corn-based finishing diet. This effect was due, in part, to

  19. Rendered-protein hydrolysates for microbial synthesis of cyanophycin biopolymer

    USDA-ARS?s Scientific Manuscript database

    Cyanophycin is a poly(arginyl-aspartate) biopolymer produced and stored intracellularly by bacteria. Cyanophycin has been proposed as a renewable replacement for petrochemical-based industrial products. An abundant source of amino acids and nitrogen such as in the form of protein hydrolysates is n...

  20. Characterising microbial protein test substances and establishing their equivalence with plant-produced proteins for use in risk assessments of transgenic crops.

    PubMed

    Raybould, Alan; Kilby, Peter; Graser, Gerson

    2013-04-01

    Most commercial transgenic crops are genetically engineered to produce new proteins. Studies to assess the risks to human and animal health, and to the environment, from the use of these crops require grams of the transgenic proteins. It is often extremely difficult to produce sufficient purified transgenic protein from the crop. Nevertheless, ample protein of acceptable purity may be produced by over-expressing the protein in microbes such as Escherichia coli. When using microbial proteins in a study for risk assessment, it is essential that their suitability as surrogates for the plant-produced transgenic proteins is established; that is, the proteins are equivalent for the purposes of the study. Equivalence does not imply that the plant and microbial proteins are identical, but that the microbial protein is sufficiently similar biochemically and functionally to the plant protein such that studies using the microbial protein provide reliable information for risk assessment of the transgenic crop. Equivalence is a judgement based on a weight of evidence from comparisons of relevant properties of the microbial and plant proteins, including activity, molecular weight, amino acid sequence, glycosylation and immuno-reactivity. We describe a typical set of methods used to compare proteins in regulatory risk assessments for transgenic crops, and discuss how risk assessors may use comparisons of proteins to judge equivalence.

  1. Detailed Dimethylacetal and Fatty Acid Composition of Rumen Content from Lambs Fed Lucerne or Concentrate Supplemented with Soybean Oil

    PubMed Central

    Alves, Susana P.; Santos-Silva, José; Cabrita, Ana R. J.; Fonseca, António J. M.; Bessa, Rui J. B.

    2013-01-01

    Lipid metabolism in the rumen is responsible for the complex fatty acid profile of rumen outflow compared with the dietary fatty acid composition, contributing to the lipid profile of ruminant products. A method for the detailed dimethylacetal and fatty acid analysis of rumen contents was developed and applied to rumen content collected from lambs fed lucerne or concentrate based diets supplemented with soybean oil. The methodological approach developed consisted on a basic/acid direct transesterification followed by thin-layer chromatography to isolate fatty acid methyl esters from dimethylacetal, oxo- fatty acid and fatty acid dimethylesters. The dimethylacetal composition was quite similar to the fatty acid composition, presenting even-, odd- and branched-chain structures. Total and individual odd- and branched-chain dimethylacetals were mostly affected by basal diet. The presence of 18∶1 dimethylacetals indicates that biohydrogenation intermediates might be incorporated in structural microbial lipids. Moreover, medium-chain fatty acid dimethylesters were identified for the first time in the rumen content despite their concentration being relatively low. The fatty acids containing 18 carbon-chain lengths comprise the majority of the fatty acids present in the rumen content, most of them being biohydrogenation intermediates of 18∶2n−6 and 18∶3n−3. Additionally, three oxo- fatty acids were identified in rumen samples, and 16-O-18∶0 might be produced during biohydrogenation of the 18∶3n−3. PMID