Maceration is an intensive forage-conditioning process that can increase field drying rates by as much as 300%. Because maceration shreds the forage and reduces its rigidity, improvements in bulk density, silage compaction, and ensiling characteristics have been observed. Macerating forage also increases the surface area available for microbial attachment in the rumen, thereby increasing forage digestibility and animal performance. Feeding trials with sheep have shown increases in DMI of 5 to 31% and increases in DM digestibility of from 14 to 16 percentage units. Lactation studies have demonstrated increases in milk production and BW gain for lactating Holstein cows; however, there is a consistent decrease in milk fat percentage when dairy cattle are fed macerated forage. In vitro studies have shown that maceration decreases lag time associated with NDF digestion and increases rate of NDF digestion. In situ digestibility studies have shown that maceration increases the size of the instantly soluble DM pool and decreases lag time associated with NDF digestion, but it may not consistently alter the rate or extent of DM and NDF digestion. (+info)
Novel genes induced during an arbuscular mycorrhizal (AM) symbiosis formed between Medicago truncatula and Glomus versiforme.
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Many terrestrial plant species are able to form symbiotic associations with arbuscular mycorrhizal fungi. Here we have identified three cDNA clones representing genes whose expression is induced during the arbuscular mycorrhizal symbiosis formed between Medicago truncatula and an arbuscular mycorrhizal fungus, Glomus versiforme. The three clones represent M. truncatula genes and encode novel proteins: a xyloglucan endotransglycosylase-related protein, a putative arabinogalactan protein (AGP), and a putative homologue of the mammalian p110 subunit of initiation factor 3 (eIF3). These genes show little or no expression in M. truncatula roots prior to formation of the symbiosis and are significantly induced following colonization by G. versiforme. The genes are not induced in roots in response to increases in phosphate. This suggests that induction of expression during the symbiosis is due to the interaction with the fungus and is not a secondary effect of improved phosphate nutrition. In situ hybridization revealed that the putative AGP is expressed specifically in cortical cells containing arbuscules. The identification of two mycorrhiza-induced genes encoding proteins predicted to be involved in cell wall structure is consistent with previous electron microscopy data that indicated major alterations in the extracellular matrix of the cortical cells following colonization by mycorrhizal fungi. (+info)
NADH-glutamate synthase in alfalfa root nodules. Genetic regulation and cellular expression.
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NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14) is a key enzyme in primary nitrogen assimilation in alfalfa (Medicago sativa L.) root nodules. Here we report that in alfalfa, a single gene, probably with multiple alleles, encodes for NADH-GOGAT. In situ hybridizations were performed to assess the location of NADH-GOGAT transcript in alfalfa root nodules. In wild-type cv Saranac nodules the NADH-GOGAT gene is predominantly expressed in infected cells. Nodules devoid of bacteroids (empty) induced by Sinorhizobium meliloti 7154 had no NADH-GOGAT transcript detectable by in situ hybridization, suggesting that the presence of the bacteroid may be important for NADH-GOGAT expression. The pattern of expression of NADH-GOGAT shifted during root nodule development. Until d 9 after planting, all infected cells appeared to express NADH-GOGAT. By d 19, a gradient of expression from high in the early symbiotic zone to low in the late symbiotic zone was observed. In 33-d-old nodules expression was seen in only a few cell layers in the early symbiotic zone. This pattern of expression was also observed for the nifH transcript but not for leghemoglobin. The promoter of NADH-GOGAT was evaluated in transgenic alfalfa plants carrying chimeric beta-glucuronidase promoter fusions. The results suggest that there are at least four regulatory elements. The region responsible for expression in the infected cell zone contains an 88-bp direct repeat. (+info)
Induction of a protective antibody response to foot and mouth disease virus in mice following oral or parenteral immunization with alfalfa transgenic plants expressing the viral structural protein VP1.
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The utilization of transgenic plants expressing recombinant antigens to be used in the formulation of experimental immunogens has been recently communicated. We report here the development of transgenic plants of alfalfa expressing the structural protein VP1 of foot and mouth disease virus (FMDV). The presence of the transgenes in the plants was confirmed by PCR and their specific transcription was demonstrated by RT-PCR. Mice parenterally immunized using leaf extracts or receiving in their diet freshly harvested leaves from the transgenic plants developed a virus-specific immune response. Animals immunized by either method elicited a specific antibody response to a synthetic peptide representing amino acid residues 135-160 of VP1, to the structural protein VP1, and to intact FMDV particles. Additionally, the immunized mice were protected against experimental challenge with the virus. We believe this is the first report demonstrating the induction of a protective systemic antibody response in animals fed transgenic plants expressing a viral antigen. These results support the feasibility of producing edible vaccines in transgenic forage plants, such as alfalfa, commonly used in the diet of domestic animals even for those antigens for which a systemic immune response is required. (+info)
Supplemental cracked corn for steers fed fresh alfalfa: I. Effects on digestion of organic matter, fiber, and starch.
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The effect of supplementation with different levels of cracked corn on the sites of OM, total dietary fiber (TDF), ADF, and starch digestion in steers fed fresh alfalfa indoors was determined. Six Angus steers (338 +/- 19 kg) fitted with cannulas in the rumen, duodenum, and ileum consumed 1) alfalfa (20.4% CP, 41.6% NDF) ad libitum (AALF); 2), 3), and 4) AALF supplemented (S) with .4, .8, or 1.2%, respectively, of BW of corn; or 5) alfalfa restricted at the average level of forage intake of S steers (RALF), in a 5 x 5 Latin square design. Total OM intake was lower (P < .01) in steers fed RALF than in those fed AALF but level of forage intake did not affect sites of OM, TDF, or starch digestion (P > .05). Forage OM intake decreased (P < .01) linearly (8,496 to 5,840 g/d) but total OM intake increased (P = .03) linearly (8,496 to 9,344 g/d) as corn increased from .4 to 1.2% BW. Ruminal apparent and true OM disappearance was not affected, but OM disappearing in the small intestine increased (P < .01) linearly with increasing levels of corn. Total tract OM digestibility (71.2 to 76.2%) and the proportion of OM intake that was digested in the small intestine (15.4 to 24.5%) increased (P < .01) linearly as corn increased. The TDF and ADF intakes decreased (P < .01) linearly as level of corn increased. Total tract TDF and ADF digestibilities were not different among treatments (average 62.9 and 57.8%, respectively). Starch intake and starch digested in the rumen and small and large intestine increased (P < .01) linearly with increasing corn level. Ruminal pH and VFA concentrations decreased and increased (P < .01), respectively, with increasing corn. Supplementation with corn increased OM intake, decreased forage OM intake, and increased the proportion of OM that was digested in the small intestine, but fiber digestion was not affected. (+info)
Supplemental cracked corn for steers fed fresh alfalfa: II. Protein and amino acid digestion.
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The effects of different levels of cracked corn on N intake, ruminal bacterial CP synthesis, and duodenal flows and small intestinal digestion of amino acids (AA) in steers fed fresh alfalfa indoors were determined. Angus steers (n = 6; average BW 338 +/- 19 kg) cannulated in the rumen, duodenum, and ileum were fed each of five diets over five periods in a Latin square design with an extra animal. Steers consumed 1) alfalfa (20.4% CP, 41.6% NDF) ad libitum (AALF); 2), 3), and 4) AALF supplemented (S) with three levels of corn (.4, .8, or 1.2% of BW, respectively), or 5) alfalfa restricted (RALF) to the average forage intake of S steers. Average N intake and duodenal flow of nonammonia N (NAN) were greater (P < .01) in S than in RALF steers. Greater duodenal flows of NAN in S compared with RALF were due to a trend toward higher (P = .06) flows of both bacterial and dietary N. Levels of corn decreased (P < .01) linearly N intake and increased (P < .01) linearly duodenal flow of NAN owing to a numerical linear increase in nonbacterial N (P = .15) with no increase in bacterial N flow. Duodenal NAN flows as percentages of N intake increased (P < .01) linearly (69.3 to 91.0%) as corn increased. Ruminal NH3 N concentration, ruminal CP degradability, and the proportion of bacterial N in duodenal NAN were decreased (P < .01) linearly as corn increased. Efficiency of net microbial CP synthesis was not affected (P > .05) by treatment (average 42.6 and 30.9 g N/kg of OM apparently or truly digested in the rumen, respectively). Small intestinal disappearance of total N and individual AA, except for threonine and lysine, and small intestinal digestibility of N and individual AA, except for methionine, histidine, and proline, increased (P < .01) linearly with level of corn and were greater (P < .01) in S than in RALF steers. Supplementing corn to steers fed fresh alfalfa reduced ruminal N losses and CP degradability and increased the duodenal flow and the small intestinal disappearance and digestibility of total N and total, essential, and nonessential AA. (+info)
Fractionation of fiber and crude protein in fresh forages during the spring growth.
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The composition of the fiber and CP of alfalfa, bromegrass, and endophyte-free and -infected tall fescue forages was compared during the spring growth from vegetative to reproductive stages. Forages were sampled from April 27 to June 6 in 1994, and from April 27 to June 11 in 1995, with 11 and 12 harvest dates, respectively. Total dietary fiber (TDF) was fractionated into insoluble and soluble fiber (SF). The CP of the forages was fractionated into nonprotein N (A), soluble CP (B1), insoluble CP that was soluble in neutral detergent (B2), CP insoluble in neutral detergent but soluble in acid detergent (B3), and CP insoluble in acid detergent (C). Effects of year, forage species, and harvest dates (day as a covariable) were included in the model. Across harvest dates, alfalfa (A) had lower (P < .01) TDF and higher (P < .01) SF concentrations than grasses (GR) (A: 49.9 and 14.4% and GR: 60.4 and 4.5% [OM basis] for TDF and SF, respectively). Alfalfa had higher (P < .01) CP (20.6% DM) than GR (15.3%). The rate of decrease in CP (% DM) across days was higher (P < .01) for bromegrass (-.4%/d) than for the other forages (-.29%/d). Fraction A (% of CP) was not different (P = .24) among forages (22.5%), but B1 was higher (P < .01) in A (17.1%) than in GR (13.2%). The B2 fraction (% of CP) was higher (P < .01) in A compared with GR (51.6 vs 45.9%, respectively). Alfalfa had lower (P < .01) B3 (3.0% of CP) than bromegrass (18.6%) and tall fescue (13.2%). Fraction C was not different (P = .23) among forages (3.8%). Fractions A, B1, and C (% of CP) did not change (P > .05) across days for all forages. Fraction B2 (% of CP) decreased across days in A (-.21%/d) but was not affected in GR. Fraction B3 (% of CP) increased (P < .05) in A (.1%/d), decreased in endophyte-infected tall fescue (-.20%/d), and did not change (P > .05) in the other forages. Crude protein and fiber composition were affected more by forage species than by maturity. The CP and NDF concentrations were more affected by maturity. Insoluble fractions but not soluble fractions of CP were affected by maturity. (+info)
Degradation of two protein sources at three solids retention times in continuous culture.
(8/896)
Effects of solids retention times (SRT) of 10, 20, and 30 h on protein degradation and microbial metabolism were studied in continuous cultures of ruminal contents. Liquid dilution rate was constant across all retention times at .12 h(-1) (8.3 h mean retention time). Two semipurified diets that contained either soybean meal (SBM) or alfalfa hay (ALFH) as the sole nitrogen source were provided in amounts that decreased as SRT was increased. Digestion coefficients for DM, NDF, and ADF increased with increasing SRT. Digestion coefficients for nonstructural carbohydrates were higher in the SBM diet than in the ALFH diet but were not affected by SRT. Protein degradation in the ALFH diet averaged 51% and was unaffected by retention time. In the SBM diet, digestion of protein was 77, 78, and 96% at 10-, 20-, and 30-h retention times, respectively. Microbial efficiency decreased with increasing SRT and was greater for the SBM than for the ALFH diet. Efficiencies ranged from 30.6 to 35.7 and 20.8 to 29.2 g of N/kg of digested DM for the SBM and ALFH diets, respectively, as SRT decreased from 30 to 10 h. The diaminopimelic acid content of the microbes increased as SRT increased, indicating that changes in microbial species occurred owing to passage rates. From these results, we concluded that the digestibility decreases associated with increased ruminal turnover rates may be less for nonstructural carbohydrates and protein than for the fiber fractions. (+info)