Effects of enzyme supplementation of a total mixed ration on microbial fermentation in continuous culture, maintained at high and low pH.
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A dual-flow continuous culture system was used to investigate the effects of pH and addition of an enzyme mixture to a total mixed ration (TMR) on fermentation, nutrient digestion, and microbial protein synthesis. A 4 x 4 Latin square design with a factorial arrangement of treatments was used, with four 9-d periods consisting of 6 d for adaptation and 3 d for measurements. Treatments were as follows: 1) high pH with control TMR, 2) high pH with TMR treated with enzyme, 3) low pH with control TMR, and 4) low pH with TMR treated with enzyme. Ranges of pH were 6.0 to 6.6 and 5.4 to 6.0 for high and low, respectively. Fermenters were fed twice daily a TMR consisting of 30% alfalfa hay, 30% corn silage, and 40% rolled corn (DM basis). The silage was milled fresh and the TMR was fed to the fermenters in fresh form (64% DM). The enzyme mixture was a commercial product of almost exclusive protease activity; it was applied daily to the fresh TMR and stored at 4 degrees C for at least 12 h before feeding. Degradability of OM, NDF, ADF, and cellulose was decreased (P < 0.05) by low pH. Hemicellulose and protein degradation were not affected by pH. Enzyme addition increased (P < 0.01) NDF degradability (by 43% and 25% at high and low pH, respectively), largely as a result of an increase in hemicellulose degradation (by 79% and 51% at high and low pH, respectively). This improvement was supported by an increase (P < 0.05) in the xylanase and cellulase activities in the liquid phase of the fermenter contents. Total VFA were decreased (P < 0.05) by low pH, but were not affected by enzyme addition. Total bacterial numbers were increased (P < 0.03) at low pH and tended (P < 0.13) to increase with enzyme addition. Cellulolytic bacteria in effluent fluid were decreased (P < 0.02) at low pH but were unaffected by enzyme addition. Despite a large increase (P < 0.001) in protease activity, protein degradation was only numerically increased by enzyme addition. Microbial protein synthesis was higher (P < 0.10) at high pH but was not affected by enzyme addition. Methane production, expressed as a proportion of total gases, was decreased (P < 0.001) at low pH but was not affected by enzyme addition. It is concluded that it is possible to adapt the continuous culture system to use fresh feeds instead of dried feeds. Overall, the results indicate that the enzyme product used in this study has a potential to increase fiber degradability without increasing methane production. (+info)
Screening of exogenous enzymes for ruminant diets: relationship between biochemical characteristics and in vitro ruminal degradation.
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With the objective of developing a rational approach for the selection of feed enzymes for ruminants, 22 commercial enzyme products were examined in terms of protein concentration, enzymic activities on model substrates, and hydrolytic capacity, the latter determined from the release of reducing sugars from alfalfa hay and corn silage. An in vitro ruminal degradation assessment was carried out using the same substrates, untreated or treated with the 22 enzyme products at 1.5 microL/g forage DM. Stepwise regressions were then performed to establish relationships between these factors. Protein concentration and enzymic activities explained at least 84% (P < 0.01) of the variation in the release of reducing sugars from alfalfa and corn silage. Alfalfa DM degradation after incubation with ruminal fluid for 18 h was positively related to xylanase activity (R2 = 0.29, P < 0.01), but the same activity was negatively related to DM degradation of corn silage (R2 = 0.19, P < 0.05). Protease activity explained a further 10% of the alfalfa DM degradation (P < 0.10). Following sequential steps involving the determination of rate and extent of DM and fiber degradation, the best candidates for alfalfa and corn silage were selected. Enzyme products effective with alfalfa hay seemed to exert part of their effect during the pretreatment period, whereas enzymes effective with corn silage worked exclusively after ruminal fluid was added. This finding suggests that different modes of action of exogenous enzymes are attacking different substrates and may partly explain enzyme-feed specificity. In alfalfa, it seems that effective enzymes work by removing structural barriers that retard the microbial colonization of digestible fractions, increasing the rate of degradation. In corn silage, effective enzymes seem to interact with ruminal enzymes to degrade the forage more rapidly, which is consistent with previous findings of synergism between exogenous and ruminal enzymes. (+info)
Composition and digestive tract retention time of ruminal particles with functional specific gravity greater or less than 1.02.
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The objective of this study was to determine composition, particle size distribution, and in vivo kinetics of ruminal particles having functional specific gravity (FSG) greater or less than FSG of particles found in the omasum and reticulum of lactating dairy cows. Particles from the reticulum and the omasal had FSG of 1.03 and 1.02, respectively. Particles from ruminal contents with FSG higher (HP) or lower (LP) than 1.02 were isolated and labeled with Er or Dy, respectively. Four ruminally cannulated, lactating Ayrshire dairy cows were fed all-grass silage (AS) or 54% grass silage:46% concentrate (SC) diets in a cross-over design trial and used to study chemical composition and ruminal and total tract kinetics of HP and LP. Labeled particles were pulse dosed into the rumen of the cows and disappearance of the markers from ruminal HP and LP pools and excretion in feces was monitored for 72 and 120 h, respectively. Fecal marker excretion data were fitted using two-compartment mathematical age-dependent/age-independent (Gn-->G1) models. Inclusion of concentrate in the diet (SC) increased (P < 0.05) apparent total tract digestibility of dietary DM, OM and N. Digestibility of fiber fractions, NDF and ADF, was lower (P < 0.01 and P < 0.05, respectively) for SC compared with AS. The heavy particles had higher (P < 0.01) indigestible NDF and lower (P < 0.01) N concentration than LP. Particles from the HP pool passed from the rumen more rapidly (P < 0.01) than particles from LP (0.044 and 0.019 h(-1), respectively). Diet had no effect on particle rate of disappearance or pool size in the rumen. Across diets, pool size of LP was consistently larger (P < 0.05) than that of HP. Diet had no effect on total tract mean retention time (MRT) of LP or HP. Total tract MRT of LP was greater (P < 0.05) than MRT of HP (59.6 vs. 49.0 h, respectively). Results from this study support the hypothesis that functional specific gravity is an important factor determining the rate of outflow and residence time of feed particles within the reticulo-rumen and total digestive tract. Our data indicate that digesta particles with functional specific gravity greater or less than 1.02 have different composition and flow characteristics. Heavier particles contain more indigestible fiber and less N and are likely depleted of substrate available for microbial fermentation, are smaller in size, and have a higher passage rate/shorter retention time in the digestive tract than lighter particles. (+info)
Kinetics of hydration and effect of liquid uptake on specific gravity of small hay and silage particles.
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Kinetics of hydration of ground hay and silage particles (2-mm screen), determined by a pycnometric technique, was best described by a two- and one-pool exponential model, respectively. Fractional rates of hydration of the large pool, detected in hay particles only, and of the small pool present in both hay and silage particles averaged .135 and .021 min-1, respectively. When hydration was complete, liquid associated with particles averaged 1.16, 1.90, and .83 g/g of insoluble DM for bromegrass hay, alfalfa hay, and alfalfa silage, respectively. Functional specific gravity, which accounts for the effect of associated gas volume, averaged 1.54, 1.46, and 1.54, but unit specific gravity, calculated to include the effect of gases and liquid of hydration, averaged 1.22, 1.14, and 1.26 for bromegrass hay, alfalfa hay, and alfalfa silage, respectively. Preservation of forage as silage not only lowered gas volume, but also reduced water-holding capacity, both of which contribute to greater unit specific gravity and faster rate of escape from the rumen. In addition, estimates of unit specific gravity of approximately 1.2 indicate that even in the absence of associated gas, hydrated forage particles would tend to escape the rumen at a slower rate than that achieved by more dense particles. (+info)
Survival of Listeria monocytogenes in wilted and additive-treated grass silage.
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Grass was field-dried to 3 different dry matter (DM) levels (200, 430 and 540 g/kg) and inoculated with 10(6)-10(7) cfu/g of a Listeria monocytogenes strain sharing a phagovar occasionally involved in foodborne outbreaks of listeriosis. Formic acid (3 ml/kg) or lactic acid bacteria (8 x 10(5)/g) with cellulolytic enzymes were applied only to forages with low and intermediate DM levels. Forages were ensiled in laboratory silos (1700 ml) and were stored at 25 degrees C for 30 or 90 days. After 90 days of storage, L. monocytogenes could not be detected in any silo, except one with the high dry matter grass without additive. After 30 days of storage, between 10(2) and 10(6) cfu L. monocytogenes/g silage were isolated from the untreated silages. Increasing the DM content from 200 to 540 g/kg did not reduce listeria counts possibly because of the lower production of fermentation acids (higher pH). In silages treated with additives, counts of L. monocytogenes were always lower than in silages without additive. In wet silages (DM 200 g/kg) both additives were effective, but in the wilted silages (DM 430 g/kg) only the bacterial additive reduced listeria counts below detection level. Listeria counts were highly correlated to silage pH (r = 0.92), the concentration of lactic acid (r = -0.80) and the pooled amount of undissociated acids (r = -0.83). (+info)
High-throughput metabolic fingerprinting of legume silage fermentations via Fourier transform infrared spectroscopy and chemometrics.
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Silage quality is typically assessed by the measurement of several individual parameters, including pH, lactic acid, acetic acid, bacterial numbers, and protein content. The objective of this study was to use a holistic metabolic fingerprinting approach, combining a high-throughput microtiter plate-based fermentation system with Fourier transform infrared (FT-IR) spectroscopy, to obtain a snapshot of the sample metabolome (typically low-molecular-weight compounds) at a given time. The aim was to study the dynamics of red clover or grass silage fermentations in response to various inoculants incorporating lactic acid bacteria (LAB). The hyperspectral multivariate datasets generated by FT-IR spectroscopy are difficult to interpret visually, so chemometrics methods were used to deconvolute the data. Two-phase principal component-discriminant function analysis allowed discrimination between herbage types and different LAB inoculants and modeling of fermentation dynamics over time. Further analysis of FT-IR spectra by the use of genetic algorithms to identify the underlying biochemical differences between treatments revealed that the amide I and amide II regions (wavenumbers of 1,550 to 1,750 cm(-1)) of the spectra were most frequently selected (reflecting changes in proteins and free amino acids) in comparisons between control and inoculant-treated fermentations. This corresponds to the known importance of rapid fermentation for the efficient conservation of forage proteins. (+info)
Technical note: A proposed method to determine the extent of degradation of a feed in the rumen from the degradation profile obtained with the in vitro gas production technique using feces as the inoculum.
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A method is proposed to determine the extent of degradation in the rumen involving a two-stage mathematical modeling process. In the first stage, a statistical model shifts (or maps) the gas accumulation profile obtained using a fecal inoculum to a ruminal gas profile. Then, a kinetic model determines the extent of degradation in the rumen from the shifted profile. The kinetic model is presented as a generalized mathematical function, allowing any one of a number of alternative equation forms to be selected. This method might allow the gas production technique to become an approach for determining extent of degradation in the rumen, decreasing the need for surgically modified animals while still maintaining the link with the animal. Further research is needed before the proposed methodology can be used as a standard method across a range of feeds. (+info)
Meat composition and palatability of Holstein and beef steers as influenced by forage type and protein source.
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This experiment determined meat composition and palatability changes resulting from feeding Holstein (HOL) and crossbred beef (XB) steers diets containing corn silage (CS) or alfalfa haylage (AH) (forage type) and soybean meal (SM) or fish meal (FM) (protein source). Fifty-nine steers (30 HOL and 29 XB) were randomly assigned to diet combinations for a 2 x 2 x 2 (breed x forage x protein) factorial arrangement. Steers were fed to a fat-constant end point (fat depth over the longissimus muscle measured by ultrasound: 1.0 cm XB, .6 cm HOL). Proximate and fatty acid analysis and sensory evaluation were conducted on a rib eye roast and steaks, respectively, removed from the left side of each carcass at ribs 9 to 12. Proximate analysis of the longissimus muscle showed no significant difference (P greater than .05) in moisture, protein, or fat content due to breed, forage, or protein treatment. Forage type had no significant effect (P greater than .05) on amount of individual fatty acids found in longissimus muscle. However, total polyunsaturated fatty acids were higher (P greater than .05) for AH than for CS-fed animals. Longissimus muscle from steers fed FM had higher palmitoleic and lower stearic acid contents (both P less than .05) than longissimus muscle from animals fed SM. Muscle from HOL had higher palmitoleic and lower stearic acid contents than that from XB steers (both P less than .05). There was no significant interaction (P greater than .05) of breed with either diet treatment for individual fatty acid contents.(ABSTRACT TRUNCATED AT 250 WORDS) (+info)