Browsing by Subject "Microbial protein synthesis"

Now showing 1 - 2 of 2
  • Thumbnail Image
    Publication
    Effects of monensin and tannin extract supplementation on methane production and other criteria of rumen fermentation in vitro and in long-term studies with sheep
    (2013) Wischer, Gerald; Rodehutscord, Markus
    Ruminants increasingly attract public concern due to their methane release and contribution to the greenhouse effect. One strategy to reduce the release of methane is to modify microbial fermentation in the rumen by the use of feed additives such as monensin and tannin extracts. However, other characteristics of fermentation including the synthesis of microbial protein may also be affected. The aim of the present studies was to provide a comprehensive evaluation of the effects of monensin and tannin extracts on ruminal fermentation and methane production. The ionophore monensin is known to increase feed efficiency in ruminants. Although the use of silages is common practice in cattle feeding, the effects of monensin on the fermentation of silages in the rumen and microbial protein synthesis are lacking. Monensin has often been described to have indirect effects on methane production resulting from its effects on feed intake, protozoa and Gram-positive bacteria. It has rarely been studied whether monensin can reduce methane production without adverse effects on other criteria of rumen fermentation. The first objective therefore was to investigate the effects of different dosages of monensin on methane production and microbial protein synthesis when supplemented to different silages in two in vitro systems (Study 1). In Experiment 1 of Study 1, 15 g of oven-dried grass silage alone or combined with a concentrate was incubated in a rumen simulation (Rusitec) over a period of 13 d to examine the effects of monensin supplementation (2 or 4 mg/d, n = 4) on the production of total gas, methane, volatile fatty acids (VFA), degradation of nutrients and microbial protein synthesis. In Experiment 2 of Study 1, different dosages of monensin (0.5, 1, 2, 6 and 10 µg) were supplemented to syringes containing 120 mg of grass silage alone, grass silage combined with concentrates, or maize silage alone. After 24 h of incubation the effects of monensin on total gas, methane and VFA production were determined. In Experiment 1 monensin inclusion to grass silage and grass silage combined with concentrate resulted in a decreased total gas, methane and acetate production, while propionate production was increased. Along with a decreased degradation of crude protein, ammonia concentration in the system was reduced. While microbial protein originating from solid associated microbes decreased with monensin inclusion, microbial protein from liquid associated microbes was increased, resulting in an increase in total microbial protein synthesis. In Experiment 2, different dosages of monensin reduced methane production in grass silage (17%), grass silage combined with concentrate (10%) and maize silage (13%) without adverse effects on total gas production. Based on these two in vitro experiments it was concluded that monensin is able to reduce methane production without a major decrease in total gas and VFA production and degradation of organic matter. Although microbial fractions were differently affected, the total microbial protein synthesis was increased upon monensin supplementation. Tannins are secondary plant compounds that are known to complex with feed and microbial proteins. Several products from this heterogeneous group have shown potential to affect rumen fermentation in vivo and, even more, in vitro, but are often accompanied by negative effects on digestibility, feed intake and microbial protein synthesis. In Study 2 of the present work, ten tannin extracts (chestnut, mimosa, myrabolan, quebracho, sumach, tara, valonea, oak, cocoa and grape seed) and four monomers of rapeseed tannin (pelargonidin, catechin, cyanidin and sinapinic acid) were screened in grass silage based diets in successive runs using the Hohenheim Gas Test. The objective was to determine the optimal dosage of each tannin extract to cause a maximal methane reduction without negative effects on total gas production. Whereas the supplementation of pelargonidin and cyanidin to grass silage did not reduce methane production; catechin and sinapinic acid reduced methane production without affecting total gas production. Except tara extract, all tannin extracts reduced methane production by 8 to 28% without adverse effects on total gas production. Based on these results, chestnut, grape seed, myrabolan, sumach and valonea extract were investigated in a second step in a Rusitec to determine their effects on degradation of nutrients, VFA and ammonia production, and particularly on microbial protein synthesis. All tannin extracts were supplemented at similar dosages of 1.5 g to 15 g of grass silage. The supplementation of chestnut resulted in the greatest decrease in methane production (63%), followed by valonea (35%), grape seed (23%), sumach (18%), and myrabolan (7%; not significantly different from the control). While 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 and organic matter was reduced by all tannin extracts, but there were no differences between tannin treatments. Crude protein degradation and ammonia production were also reduced by tannin extract supplementation. Microbial protein synthesis and its efficiency were not affected by tannin supplementation, which indicates that a reduction in methane production due to tannin extract supplementation is possible without negatively affecting microbial protein synthesis. Chestnut and valonea extract had the greatest potential in reducing methane production without negative effects on rumen fermentation of grass silage and microbial protein synthesis. Therefore, these tannin extracts were investigated for their long-term effects in sheep (Study 3). In Experiment 1 of Study 3, sheep receiving the control, chestnut or valonea treatment (each n = 4) were fed 842 g/d of hay (fresh weight). The animals on the control treatment also received 464 g/d of concentrate, and animals on the tannin treatments received the same amount of concentrate but were also fed 20 g of the respective tannin extract. Following initiation of tannin feeding, methane release from sheep was measured in 23.5 h intervals in respiration chambers on day 1, 8, 15, 29, 57, 85, 113, 148, and 190. In three balances periods faeces and urine were collected for 6 and 3 days, respectively. Effects on nutrient digestibility, nitrogen and energy metabolism were evaluated, with microbial protein synthesis estimated from the urinary excretion of purine derivatives. Based on the results of Experiment 1, a second experiment was conducted four month after the start of Experiment 1. Experiment 2 had the same study design and data collected, but the dosage of tannin extracts was doubled compared to Experiment 1 (0.9 vs. 1.7 g tannin extract/kg body weight) and the duration was shorter (85 days). Hay and concentrates used in both experiments were also evaluated using the Hohenheim Gas for their effects on total gas and methane production. In both experiments, methane release was not significantly reduced by tannin extract supplementation when analysed over the whole experimental period. In Experiment 1 the supplementation of chestnut extract on day 190 resulted in a reduced methane release. In both experiments, on day 1 a numeric reduction in methane release for the tannin treatments was observed, with a greater reduction recorded for the higher dosage used in Experiment 2. This trend disappeared by day 57. In the third balance period of Experiment 1, digestibility of dry and organic matter was reduced by tannin supplementation. The digestibility of crude protein was reduced in both experiments, whereas the digestibility of fibre fractions was not influenced. In both experiments a long-lasting shift in nitrogen excretion from urine to faeces was observed, which occurred to a greater extent in Experiment 2. The urinary excretion of purine derivatives was not significantly affected by tannin supplementation, indicating that the microbial protein synthesis was not altered in either experiment. The in vitro methane production was reduced for concentrates containing tannin extracts, but it was not significantly affected when concentrates were incubated with hay. It is concluded that monensin added to different silages caused a decrease in methane production without affecting total gas production but with an increased microbial protein synthesis. Nine of the ten considered tannin extracts and two tannin monomers decreased methane production without affecting total gas production. The Rusitec study confirmed the great potential of chestnut and valonea extract to reduce methane production without negative effects on microbial protein synthesis. However, neither chestnut nor valonea extract reduced the methane release in sheep when fed over a longer period of time. It is assumed, that rumen microbes adapted to the tannin dosages in terms of methane release but not nitrogen metabolism, as there were long-lasting effects on nitrogen excretion. The shift in nitrogen excretion can have a positive effect on the environment due to the reduced potential of ammonia emission from the urine. Both in vitro systems used in the present studies showed effects of tannin extracts that were considerably different from those observed in sheep. The monomers investigated in the present study are the basic units of condensed tannins, whereas the tannin extracts selected in vitro only contain hydrolysable tannins. It is possible that monomers of chestnut and valonea extract may reduce methane production, whereas higher dosages of these tannin extracts cause negative effects on feed intake, digestibility and microbial protein synthesis. Further investigations should focus systematically on the transfer of in vitro studies to estimate in vivo responses. Therefore, a parallel implementation of different in vitro and respiration studies would be of great value.
  • Thumbnail Image
    Publication
    Fermentations- und Syntheseleistung der mikrobiellen Gemeinschaft des Pansens in vitro bei Variation der Grobfutter- und Stickstoffquellen
    (2017) Zuber, Karin Helga Renate; Rodehutscord, Markus
    In the first part of this doctoral thesis five batches of maize silage (MS), five batches of grass silage (GS) and three batches of alfalfa silage were incubated in the Hohenheim gas test. The variation of silages based on in vitro gas production kinetics and ammonia-nitrogen-concentration (NH3-N-concentration) in the mixture of rumen liquid and buffer solution over time was determined. For this purpose, 10 glass syringes per silage batch were used per experimental run. 3 glass syringes were used to determine the gas volume over 72 hours. The remaining 7 glass syringes were removed from the incubator at 7 time points and the NH3-N-concentration in the mixture of rumen liquid and buffer solution was determined. Upon the incubation of the 13 silages both silage species and batch had an influence on the potential gas production and on the rate constant of gas production. The determined potential gas production was between 62.5–74.2, 56.0–64.9 and 39.9–59.6 mL/200 mg organic matter (OM) for MS, GS and alfalfa silages. The rate constant of gas production amounted to 5.5–7.3, 3.8–7.1 and 5.0–7.7 %/h for MS, GS and alfalfa silages. Both silage species and batch as well as the time point and their interactions had an influence on the NH3-N-concentration in the mixture of rumen liquid and buffer solution. In the second part of this work one MS and one GS were incubated in the rumen simulation Rusitec. The influence of the forage source without supplementation of concentrates on the NH3-N-concentration in fermenter liquids over time and the fermentation and synthesis characteristics of the ruminal microbial community were investigated in vitro. Degradation of nutrients, gas, methane and short chain fatty acid (SCFA) production as well as NH3-N in effluent and microbial protein synthesis (MPS) were measured. The NH3-N-concentration in fermenter liquids was determined at different time points within two periods. Upon the incubation of GS, degradation of OM and fibre fractions, amount of NH3-N in the effluent as well as MPS and its efficiency (EMPS) was higher than with incubation of MS. Degradation of crude protein (CP) and total amount of SCFA were unaffected by silage. N-efficiency was higher with incubation of MS than with incubation of GS. During period 1, NH3-N-concentration in fermenter liquids increased for all treatments within the first 24 hours and was not different between the treatments. For GS, NH3-N-concentration subsequently continued to rise up to a maximum value at the last time point of measurement in period 1. NH3-N-concentrations in fermenter liquids in period 2 remained on a relatively constant level for MS and GS, differing between the two silages at all five time points of measurement. Mean NH3-N-concentration in fermenter liquids measured in period 2 corresponded in level with NH3-N-concentration determined in the effluent of both silages. In the third part of this work, the influence of different N-supplements to MS compared to GS on fermentation and synthesis characteristics of the ruminal microbial community in vitro was investigated. GS and MS were incubated in a Rusitec, the latter being either unsupplemented or supplemented with urea, pea protein, pea peptone or a mixture of amino acids to adjust N-content of MS to that of GS. The NH3-N-concentration in fermenter liquids was determined 0, 2, 4, 12 and 24 hours after changing the feed bag on day 12. Results concerning degradation of OM, CP and N-free extracts showed a positive influence of N-supplementations except for MS+pea protein. Furthermore, degradation of detergent fibres were partially improved through N-supplementations. The values of MPS and EMPS were enhanced through all N-supplementations. Thereby supplementation of urea and pea peptone to MS resulted in the largest increase in EMPS. However, through none of the N-supplements the level of GS in EMPS could be achieved. The determined course of NH3-N-concentration in fermenter liquids was largely similar between the treatments. Variation in nutrient composition of MS, GS and alfalfa silages were reflected in a large variation both in gas production kinetics and curve shape of NH3-N-concentration in the mixture of rumen liquid and buffer solution. Upon the sole incubation of MS and GS in the Rusitec, GS promoted MPS and EMPS stronger than MS. Supplementation of MS with different N-sources resulted in an increase in MPS and EMPS compared to MS without N-supplementation. Thus the assumption of an insufficient N-supply of ruminal microbes during the sole incubation of MS in vitro was confirmed. However, through none of the N-supplementations level of GS in EMPS could be achieved.