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Effect of white rot basidiomycetes on chemical composition and in vitro digestibility of oat straw and alfalfa stems.

Five white rot basidiomycetes were evaluated for their potential to improve ruminal degradation of oat straw and alfalfa stems. Phanerochaete chrysosporium (PC), Scytinostroma galactinum (SG), Phlebia tremellosa (PT), Phellinus pini (PP), and Pholiota mutabilis (PM) were incubated on oat straw and alfalfa stems for 30 d at 28 degrees C and 90% relative humidity. Detergent fiber and total fiber components (neutral sugars, uronic acids, Klason lignin [KL], and ester- and ether-linked non-core lignin phenolics), core lignin nitrobenzene oxidation products, and IVDMD were determined. Electron microscopy of KMnO4-stained and cellulase/colloidal gold-labeled sections was used to monitor fungal activity. Large losses of DM were noted for all fungal species on both substrates. Lignin (KL and ADL) was removed (P less than .05) from oat straw by PC and PT treatment, but no net loss of lignin was observed for fungal treatment of alfalfa stems. Cell-wall polysaccharides were removed from both substrates by fungal activity. Only PC increased (P less than .05) IVDMD of oat straw, and SG, PT, PP, and PC treatment decreased (P less than .05) IVDMD of alfalfa stems, presumably because the fungi removed the most readily fermentable polysaccharides. Transmission electron microscopy using KMnO4 staining showed a nonselective white rot attack. Cytochemical studies using colloidal gold-labeled exo- and endocellulases were used to map the location of cellulose in the cell wall before and after decay by the white rot fungi. All the white rot fungi tested had eroded and thinned cell walls. Residual cell walls were well-labeled; both endo- and exocellulose-colloidal gold identified the cellulosic wall material that remained.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗

Characteristics of plant cell walls affecting intake and digestibility of forages by ruminants.

Even under the intensive concentrate feeding systems of ruminant animal production in the United States, forages continue to represent the single most important feed resource. Cell-wall concentration and digestibility limit the intake potential and energy availability of forage crops in beef and dairy production. Identification of cell-wall characteristics that should be targets of genetic modification is required if plant breeders and molecular biologists are to successfully improve forages for livestock feeding. As the forage plant cell develops, phenolic acids and lignin are deposited in the maturing cell wall in specific structural conformations, and in a strict developmental sequence. Lignin is the key element that limits cell-wall digestibility, but cross-linkage of lignin and wall polysaccharides by ferulic acid bridges may be a prerequisite for lignin to exert its affect. Lignin composition and p-coumaric acid in the wall are less likely to affect digestibility. Voluntary intake of forages is a critical determinant of animal performance and cell-wall concentration is negatively related to intake of ruminants consuming high-forage diets. Cell walls affect intake by contributing to ruminal fill. A simple model of cell-wall digestion and passage in which ruminal fill is a function of rates of digestion and passage, as well as the indigestible fraction of the cell-wall indicates that cell-wall concentration and rate of passage are the most critical parameters determining ruminal fill. Plant factors that affect rate of passage include those that affect particle size reduction by chewing and those that affect particle buoyancy in the rumen. The latter is primarily affected by 1) the ability of the particulate matter to retain gases, which is probably related to plant anatomy and rate of digestion of the plant tissue, and 2) the rate at which the gas is produced, which is affected by the potentially digestible fraction of the particulate matter and the rate of digestion of this fraction. Increasing rate of digestion should increase rate of passage by diminishing the gas produced and increasing density over time. A reduction in the indigestible cell-wall fraction is beneficial because this will decrease fill and increase digestibility. Animal production and economic benefits from reduced cell-wall concentration and increased digestibility are significant. Because of the high cell-wall concentration and large digestible cell-wall fraction of grasses, reduction in cell-wall concentration would probably be of greater value than improving digestibility in these species. Legumes represent the opposite situation and may benefit more from improvements in the digestibility of their cell walls.

Animal Feed↗

Elucidation of factors associated with the maturity-related decline in degradability of big bluestem cell wall.

Big bluestem (Andropogon gerardii) forage samples were collected from three ungrazed, annually burned pastures at 38, 58, and 97 d after burning. Cell wall material received five treatments: chlorite delignification, chlorite delignification plus alkali extraction, NaOH, NaOCH3 in methanol, or NaBH4. Untreated and treated cell walls were analyzed for carbohydrate composition (glucose, xylose, arabinose, galactose, and uronic acids), acetyl bromide lignin, acid detergent lignin, alkali-labile phenolics (p-coumaric and ferulic acids), acetyl groups, and 24- and 72-h in vitro degradabilities of neutral monosaccharides. A number of compositional features, notably concentrations of xylose, core lignin as measured by acetyl bromide lignin, alkali-labile phenolics, and acetyl groups, were well related to the decline in cell wall degradability that occurred with increasing maturity of big bluestem. p-Coumaric acid increased with increasing maturity to a greater extent than did ferulic acid. Acid detergent lignin was not well related to degradability of the cell wall for either the untreated or chemically treated cell walls. Chemical treatments failed to identify any particular cell wall component as being most inhibitory. However, all treatments improved in vitro degradability of the carbohydrate fraction, indicating that components contributing to the undegradability of big bluestem cell wall are sensitive to chemical alteration. Treatments involving alkali were most effective for improving degradability of big bluestem cell walls.

Aging↗

Pretreatment of sugar cane bagasse for enhanced ruminal digestion.

Crop residues, such as sugar cane bagasse (SCB), have been largely used for cattle feeding. However, the close association that exists among the three major plant cell-wall components, cellulose, hemicellulose, and lignin, limits the efficiency by which ruminants can degrade these materials. Previously, we have shown that pretreatment with 3% (w/w) phosphoric acid, under relatively mild conditions, increased considerably the nutritional value for SCB. However, in this preliminary study, pretreated residues were not washed prior to in situ degradability assays because we wanted to explore the high initial solvability of lowmol-wt substances that were produced during pretreatment. We have now studied the suitability of water-and/or alkali-washed residues to in situ ruminal digestion. Alkali washing increased substrate cellulose content by removing most of the lignin and other residual soluble substances. As a result the ruminal degradability of these cleaner materials had first-order rate constants five times higher than those substrates with higher lignin content (e.g., stem-exploded bagasse). However, alkali washing also increased the time of ruminal lag phase of the cellulosic residue, probably because of hemicellulose and/or lignin removal and to the development of substrates with higher degree of crystallinity. Therefore, longer lag phases appear to be related to low microbial adherence after extensive water and alkali extraction, as Novell as to the slower process of cellulase induction during ruminal growth. The kinetic data on ruminal digestion were shown to be very well adjusted by a nonlinear model. Although pretreatment enhances substrate accessibility, the occurrence of an exceedingly high amount of lignin byproducts within the pretreated material reduces considerably its potential degradability.

Alkalies↗

Quantitative trait loci for cell-wall components in recombinant inbred lines of maize (Zea mays L.) I: stalk tissue.

Maize silage is a significant energy source for animal production operations, and the efficiency of the conversion of forage into animal mass is an important consideration when selecting cultivars for use as feed. Fiber and lignin are negatively correlated with digestibility of feed, so the development of forage with reduced levels of these cell-wall components (CWCs) is desirable. While variability for fiber and lignin is present in maize germplasm, traditional selection has focused on the yield of the ear rather than the forage quality of the whole plant, and little information is available concerning the genetics of fiber and lignin. The objectives of this study were to map quantitative trait loci (QTLs) for fiber and lignin in the maize stalk and compare them with QTLs from other populations. Stalk samples were harvested from 191 recombinant inbred lines (RILs) of B73 (an inbred line with low-to-intermediate levels of CWCs) x De811 (an inbred line with high levels of CWCs) at two locations in 1998 and one in 1999 and assayed for neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL). The QTLs were detected on nine chromosomes, mostly clustered in concordance with the high genetic correlations between NDF and ADF. Adjustment of NDF for ADF and ADF for ADL revealed that most of the variability for CWCs in this population is in ADF. Many of the QTLs detected in this study have also been detected in other populations, and several are linked to candidate genes for cellulose or starch biosynthesis. The genetic information obtained in this study should be useful to breeding efforts aimed at improving the quality of maize silage.

Breeding↗

Unpolluted fractionation of wheat straw by steam explosion and ethanol extraction.

An unpolluted process of wheat straw fractionation by steam explosion coupled with ethanol extraction was studied. The wheat straw was steam exploded for 4.5 min with moisture of 34.01%, a pressure of 1.5 MPa without acid or alkali. Hemicellulose sugars were recovered by water countercurrent extraction and decolored with chelating ion exchange resin D412. The gas chromatography (GC) and high-performance liquid chromatography (HPLC) analysis results indicated that there were organic acids in the hemicellulose sugars and the ratio of monosaccharides to oligosaccharides was 1:9 and the main component, xylose, was 85.9% in content. The total recovery rate of hemicellulose was 80%. Water washed materials were subsequently extracted with ethanol. The optimum extraction conditions in this work were 40% ethanol, fiber/liquor ratio 1:50 (w/v), severity log(R)=3.657 (180 degrees C for 20 min), 0.1% NaOH. The lignin yield was 75% by acid precipitation and 85% ethanol solvent was recovered. The lignin was purified using Björkman method. Infrared spectrometry (IR) results indicated that the lignin belonged to GSH (guaiacyl (G) syringyl (S) and p-hydroxyphenyl (H)) lignin and its purity rate reached 85.3%. The cellulose recovery rate was 94% and the results of electron spectroscopy for chemical analysis (ESCA) and infrared spectrometry (IR) showed that hemicellulose and lignin content decreased after steam explosion and ethanol extraction.

Chemical Fractionation↗

Wet strength improvement of unbleached kraft pulp through laccase catalyzed oxidation.

Previous investigations have shown that laccase catalyzed oxidation of lignin containing wood fibers can enhance the strength of medium density fiberboards. In the present work it was investigated if laccase treatment had any impact on the tensile strength of a high yield unbleached kraft pulp. Treatment with laccase alone had only a very little effect on the wet strength of the pulp, whereas addition of lignin rich extractives increased the wet strength after the enzyme treatment significantly. A mediated oxidation gave a similar improvement of the wet tensile strength although no lignin was added to the fiber suspension. Furthermore, it was found that a heat treatment combined with a mediated oxidation gave a higher improvement in wet tensile strength than could be accounted for by the individual treatments. No change in dry tensile strength from the laccase treatment was observed. It is suggested that the observed improvement in wet tensile strength is related to polymerization of lignin on fibers in the hand sheet and/or coupling of phenoxy radicals on lignin associated to adjacent fibers. For the different mediators studied, a correlation was found between oxygen consumption upon mediated oxidation and generation of wet strength in the pulp.

Journal Article↗

Ozonation Products of Organosolvolytic Extracts from Vegetal Materials.

The ozonation of lignin in an organic solution was used to produce oxyaromatic compounds. Corn (Zea mays) stalk and poplar (Populus deltoides) wood lignin solutions (organosolvolytic juices) obtained from their respective autohydrolyzed materials and an organosolv lignin were treated with ozone, and the time course trend of the identified reaction products was studied. GC/MS and GC/FID were used to identify and quantify (internal standard method) the reaction products. The following acids were identified: glycolic, oxalic, malic, glyceric, p-hydroxybenzoic, and vanillic. In addition, p-hydroxybenzaldehyde, vanillin, syringaldehyde, and hydroquinone were also identified. All the oxyaromatic aldehydes reached maximum yields at short reaction times (between 1 and 8 min). These results were used to determine the ratio between the different phenylpropane units of their constitutive lignins. The ratio of p-hydroxyphenyl:guaiacyl:syringyl units for corn stalk, poplar wood, and organosolv lignins was 5.3:2:1, 0.1:1:1.1, and 0:1:1.1, respectively.

Journal Article↗

Caffeoyl-coenzyme A 3-O-methyltransferase enzyme activity, protein and transcript accumulation in flax (Linum usitatissimum) stem during development.

Flax (Linum usitatissimum) is a commercially important fiber crop in Europe. Lignification of its phloem fibers, although weak, causes a decrease in their commercial quality. In flax, fiber lignin mainly consists of guaiacyl (G) units in contrast to the mixed guaiacyl-syringyl (G-S) lignin type occurring in xylem fibers. G lignins are reported as more condensed polymers due to a higher frequency of 5-5 linkages, whereas the deposition of syringyl end groups in lignins increases the proportion of alky-aryl ether linkages as beta-O-4-bonds. The type of linkages within a lignin polymer depends on the methylation of either the 3-hydroxyl groups or both 3-OH and 5-OH groups, which is controlled by two enzymes: caffeate 3-O-methyltransferase (COMT) and caffeoyl-coenzyme A 3-O-methyltransferase (CCoAOMT). First, we measured the in vitro activity of both OMTs in the flax stem tissues during stem development. CCoAOMT activity varied in the same way as COMT, i.e. increased gradually with stem maturity, from the top to the bottom of the stem, was maximum at the flowering stage and was lower, but still scorable, in the outer fiber-bearing tissues than in the xylem cells. In a second step, we focused our studies on the characterization of CCoAOMT in order to understand the implication of this enzyme in the lignification of flax fiber cells. CCoAOMT activity appeared to be associated with the accumulation of an acidic 33-kDa polypeptide identified as a CCoAOMT after immunological cross-reactivity with a poplar CCoAOMT and microsequencing. The differential accumulation of the CCoAOMT protein was confirmed by immunolocalization on tissue prints and correlated with that of the transcripts, suggesting a transcriptional regulation of CCoAOMT in the flax stem.

Journal Article↗

Apoplastic Peroxidases and Lignification in Needles of Norway Spruce (Picea abies L.).

The objective of the present study was to investigate the correlation of soluble apoplastic peroxidase activity with lignification in needles of field-grown Norway spruce (Picea abies L.) trees. Apoplastic peroxidases (EC 1.11.1.7) were obtained by vacuum infiltration of needles. The lignin content of isolated cell walls was determined by the acetyl bromide method. Accumulation of lignin and seasonal variations of apoplastic peroxidase activities were studied in the first year of needle development. The major phase of lignification started after bud break and was terminated about 4 weeks later. This phase correlated with a transient increase in apoplastic guaiacol and coniferyl alcohol peroxidase activity. NADH oxidase activity, which is thought to sustain peroxidase activity by production of H2O2, peaked sharply after bud break and decreased during the lignification period. Histochemical localization of peroxidase with guaiacol indicated that high activities were present in lignifying cell walls. In mature needles, lignin was localized in walls of most needle tissues including mesophyll cells, and corresponded to 80 to 130 [mu]mol lignin monomers/g needle dry weight. Isoelectric focusing of apoplastic washing fluids and activity staining with guaiacol showed the presence of strongly alkaline peroxidases (isoelectric point [greater than or equal to] 9) in all developmental stages investigated. New isozymes with isoelectric points of 7.1 and 8.1 appeared during the major phase of lignification. These isozymes disappeared after lignification was terminated. A strong increase in peroxidase activity in autumn was associated with the appearance of acidic peroxidases (isoelectric point [less than or equal to] 3). These results suggest that soluble alkaline apoplastic peroxidases participate in lignin formation. Soluble acidic apoplastic peroxidases were apparently unrelated to developmentally regulated lignification in spruce needles.

Journal Article↗

Relative contributions of bacteria and fungi to rates of degradation of lignocellulosic detritus in salt-marsh sediments.

Specifically radiolabeled [C-lignin]lignocellulose and [C-polysaccharide]lignocellulose from the salt-marsh cordgrass Spartina alterniflora were incubated with an intact salt-marsh sediment microbial assemblage, with a mixed (size-fractionated) bacterial assemblage, and with each of three marine fungi, Buergenerula spartinae, Phaeosphaeria typharum, and Leptosphaeria obiones, isolated from decaying S. alterniflora. The bacterial assemblage alone mineralized the lignin and polysaccharide components of S. alterniflora lignocellulose at approximately the same rate as did intact salt-marsh sediment inocula. The polysaccharide component was mineralized twice as fast as the lignin component; after 23 days of incubation, ca. 10% of the lignin component and 20% of the polysaccharide component of S. alterniflora lignocellulose were mineralized. Relative to the total sediment and bacterial inocula, the three species of fungi mediated only very slow mineralization of the lignin and polysaccharide components of S. alterniflora lignocellulose. Experiments with uniformly C-labeled S. alterniflora material indicated that the three fungi and the bacterial assemblage were capable of degrading the non-lignocellulosic fraction of S. alterniflora material, but only the bacterial assemblage significantly degraded the lignocellulosic fraction. Our results suggest that bacteria are the predominant degraders of lignocellulosic detritus in salt-marsh sediments.

Journal Article↗

Characterization of palo podrido, a natural process of delignification in wood.

Chemical and morphological changes of incipient to advanced stages of palo podrido, an extensively delignified wood, and other types of white rot decay found in the temperate forests of southern Chile were investigated. Palo podrido is a general term for white rot decay that is either selective or nonselective for the removal of lignin, whereas palo blanco describes the white decayed wood that has advanced stages of delignification. Selective delignification occurs mainly in trunks of Eucryphia cordifolia and Nothofagus dombeyi, which have the lowest lignin content and whose lignins have the largest amount of beta-aryl ether bonds and the highest syringyl/guaiacyl ratio of all the native woods included in this study. A Ganoderma species was the main white rot fungus associated with the decay. The structural changes in lignin during the white rot degradation were examined by thioacidolysis, which revealed that the beta-aryl ether-linked syringyl units were more specifically degraded than the guaiacyl ones, particularly in the case of selective delignification. Ultrastructural studies showed that the delignification process was diffuse throughout the cell wall. Lignin was first removed from the secondary wall nearest the lumen and then throughout the secondary wall toward the middle lamella. The middle lamella and cell corners were the last areas to be degraded. Black manganese deposits were found in some, but not all, selectively delignified samples. In advanced stages of delignification, almost pure cellulose could be found, although with a reduced degree of polymerization. Cellulolytic enzymes appeared to be responsible for depolymerization. A high brightness and an easy refining capacity were found in an unbleached pulp made from selectively delignified N. dombeyi wood. Its low viscosity, however, resulted in poor resistance properties of the pulp. The last stage of degradation (i.e., decomposition of cellulose-rich secondary wall layers) resulted in a gelatinlike substance. Ultrastructural and chemical analyses of this substance showed the matrix to have no microfibrillar structure characteristic of woody cell walls but to still be rich in glucan.

Journal Article↗

Comparison of extracellular peroxidase- and esterase-deficient mutants of Streptomyces viridosporus T7A.

Peroxidase-deficient mutants of the lignin-degrading bacterium Streptomyces viridosporus T7A were screened for their production of acid-precipitable polymeric lignin, extracellular peroxidases and esterases, and immunoreactivities against a polyclonal antibody produced against electrophoretically purified peroxidase isoform P3 of wild-type S. viridosporus. The mutants showed diminished abilities to solubilize lignin and produce acid-precipitable polymeric lignin. Their peroxidase activities were decreased, and their esterase production patterns were altered. Western immunoblots demonstrated that the mutants produced proteins immunologically reactive with the antibody, but with different mobilities from those of wild-type proteins. These findings confirm a direct role for peroxidases in lignin solubilization. They also indicate a possible role for esterases.

Esterases↗

Lignocellulose Degradation during Solid-State Fermentation: Pleurotus ostreatus versus Phanerochaete chrysosporium.

Lignocellulose degradation and activities related to lignin degradation were studied in the solid-state fermentation of cotton stalks by comparing two white rot fungi, Pleurotus ostreatus and Phanerochaete chrysosporium. P. chrysosporium grew vigorously, resulting in rapid, nonselective degradation of 55% of the organic components of the cotton stalks within 15 days. In contrast, P. ostreatus grew more slowly with obvious selectivity for lignin degradation and resulting in the degradation of only 20% of the organic matter after 30 days of incubation. The kinetics of C-lignin mineralization exhibited similar differences. In cultures of P. chrysosporium, mineralization ceased after 18 days, resulting in the release of 12% of the total radioactivity as CO(2). In P. ostreatus, on the other hand, 17% of the total radioactivity was released in a steady rate throughout a period of 60 days of incubation. Laccase activity was only detected in water extracts of the P. ostreatus fermentation. No lignin peroxidase activity was detected in either the water extract or liquid cultures of this fungus. 2-Keto-4-thiomethyl butyric acid cleavage to ethylene correlated to lignin degradation in both fungi. A study of fungal activity under solid-state conditions, in contrast to those done under defined liquid culture, may help to better understand the mechanisms involved in lignocellulose degradation.

Journal Article↗

Heterogeneity and regulation of manganese peroxidases from Phanerochaete chrysosporium.

Lignin and Mn peroxidases are two families of isozymes produced by the lignin-degrading fungus Phanerochaete chrysosporium under nutrient nitrogen or carbon limitation. We purified to homogeneity the three major Mn peroxidase isozymes, H3 (pI = 4.9), H4 (pI = 4.5), and H5 (pI = 4.2). Amino-terminal sequencing of these isozymes demonstrates that they are encoded by different genes. We also analyzed the regulation of these isozymes in carbon- and nitrogen-limited cultures and found not only that the lignin and Mn peroxidases are differentially regulated but also that differential regulation occurs within the Mn peroxidase isozyme family. The isozyme profile and the time at which each isozyme appears in secondary metabolism differ in both nitrogen- and carbon-limited cultures. Each isozyme also responded differently to the addition of a putative inducer, divalent Mn. The stability of the Mn peroxidases in carbon- and nitrogen-limited cultures was also characterized after cycloheximide addition. The Mn peroxidases are more stable in carbon-limited cultures than in nitrogen-limited cultures. They are also more stable than the lignin peroxidases. These data collectively suggest that the Mn peroxidase isozymes serve different functions in lignin biodegradation.

Amino Acid Sequence↗

Assessment of microencapsulated formulations for improved residual activity of Bacillus thuringiensis.

Bacillus thuringiensis Berliner is a highly efficacious bioinsecticide used to control lepidopteran pests in the field. Unfortunately, it has limited residual activity on plants because sunlight inactivates spores and crystals and they can be washed off by rain. To minimize loss of activity, formulations must contain UV protectants, stickers, or both. We tested approximately 80 formulations and determined optimal combinations of ingredients and spray drying conditions for improving B. thuringiensis residual activity after simulated rain and simulated sunlight. B. thuringiensis stability, after simulated sunlight (xenon light/8 h) and rain (5 cm/50 min), was improved using formulations based on lignin, corn flours, or both, with up to 20% of the active ingredient, when compared with technical powder or Dipel 2x in laboratory assays. Two formulations, made with corn flours or lignin + pregelatinized corn flour (PCF), killed 51.6 and 75.3% of Ostrinia nubilalis (Hübner) neonates after rain, respectively, versus 27% for technical powder. When the insecticidal activity was tested after simulated sunlight, corn flour-based formulations killed 78.5% of test larvae, and the lignin + PCF formulation killed 70.4%, in contrast to technical powder which caused an average of 29% mortality. Formulations made with Dipel 2x rather than technical powder, caused 62.5% mortality (corn flour-based formulations), and 72.3% mortality (lignin + PCF), versus 53.4% for Dipel 2x after rain. When tested after simulated sunlight, formulations killed 95% of the larvae (average of both formulations) versus 82% for Dipel 2x. In a field test, formulations were applied to cabbage and insecticidal activity was determined against Trichoplusia ni (Hübner) neonates exposed to treated leaves. Insecticidal activity of the corn flour-based formulations was comparable to Dipel 2x for 4 d after treatment, but was significantly better than Dipel 2x 7 d after application. A lignin and PCF-based formulation showed significantly higher residual activity than Dipel 2x, 4 and 7 d after application.

Animals↗

The effect of physical form of orchardgrass hay on the passage of particulate matter through the rumen of sheep.

Four Texel wethers (60 to 64 kg) fitted with ruminal and duodenal cannulas were used to study the kinetics of particulate matter in the rumen and the series of processes involved in their selection and passage. They were fed, in eight equal meals, 1,200 g of a mixture of a chopped orchardgrass hay and ground (8-mm screen) and pelleted orchardgrass hay in 90/10, 50/50, 30/70, or 10/90 ratios, according to a 4 x 4 Latin square design. The particle size distributions in feed, chewed feed, and ruminal, reticular, and duodenal digesta were determined by a wet-sieving procedure. Indigestible lignin was used as an internal marker to trace the passage of particles through the rumen. Digesta flow measurement was performed using the double-marker technique. We used a three-pool model, which partitions particles through the large, medium, and small particle pools, to determine the passage of lignin through those pools. Particle pool sizes and rumen and pool mean retention times (MRT) of lignin and of the rumen MRT of an ideal marker introduced separately in each pool were corrected for the "filter bed" effect. Grinding and pelleting of hay decreased the MRT of the indigestible lignin pool in the rumen. Particle MRT decreased and then reached a plateau with increased proportion of ground/pelleted hay in the diet. The diet with a ratio of 50/50 of chopped and ground/pelleted hay was the most favorable for the exit of particles from the rumen because of both a higher outflow rate from the rumen of particles eligible to exit and a sufficient comminution rate of larger particles to supply particles that were able to pass. For all diets, the large-particle comminution rate was always higher than the small-particle outflow rate, indicating that comminution was not the limiting step for passage. These results were the consequence of the curvilinear and opposite evolutions of both the particulate lignin pool in, and outflow from, the rumen. Those results contribute to an improved explanation of the mechanisms involved in the outflow of particles from the rumen.

Animal Feed↗

Biological activity of phenolic compounds. Hepatic cytochrome P-450, cytochrome b5, and NADPH cytochrome c reductase in chicks and rats fed phenolic monomers, polymers, and glycosides.

Eight experiments were conducted to determine effects of a phenolic polymer (Kraft wood lignin, Indulin), phenolic glycosides (cane molasses and wood molasses), and phenolic monomers (vanillin, vanillic acid, ferulic acid, and p-coumaric acid) on liver cytochromes P-450, cytochrome b5, and NADPH cytochrome c reductase in chicks and rats. Chicks fed 6.0% lignin had a higher (P less than 0.01) cytochromes P-450 content than did chicks fed 0% fiber, 6.0% wood cellulose (Solka Floc), or 6.0% arenaceous flour. NADPH cytochrome c reductase activity was not affected by treatment. Chicks fed 12.0% wood molasses had a higher (P less than 0.06) cytochromes P-450 level than did chicks fed 0% fiber or 6.0% wood molasses. Cane molasses incorporated at both 6.0 and 12.0% of the diet induced (P less than 0.05) cytochromes P-450 content over those of control-fed birds. Chicks fed 6.0% lignin, with or without antibiotic (bacitracin:neomycin sulfate, 2:1), had a higher (P less than 0.01) cytochromes P-450 level than did chicks fed control diets, with or without antibiotic. Additionally, chicks fed 6.0% lignin had lower (P less than 0.01) intestinal diaminopimelic acid (DAP) levels than did chicks fed 0% fiber. Rats fed 0% fiber, 6.0% wood cellulose, 6.0% arenaceous flour, or 6.0% lignin exhibited no difference in cytochrome level or activity among treatments. Chicks fed 0.5% vanillin, 0.5% vanillic acid, 0.5% ferulic acid, or 0.5% p-coumaric acid had comparable cytochromes level and activity compared with chicks fed no phenolics. Chicks fed 0.5% p-coumaric acid had lower (P less than 0.05) rates of gain than did chicks fed control or other phenolic-containing diets. Rats fed these phenolics had similar cytochromes P-450 content among treatments.

Age Factors↗