In vitro antimicrobial efficacy of Cannabis sativa L. (hemp) crude oil extract on Fusobacterium necrophorum of bovine origin.

Fusobacterium necrophorum commonly causes liver abscesses in grain-finished beef, which is mitigated using antibiotics in feed. However, microbial resistance development necessitates exploration of antibiotic alternatives. Here we report the antimicrobial efficacy of Cannabis sativa L.

Reduction in Rumen Tetracycline-Insensitive Bacteria during a Grain Challenge Using the Isoflavone Biochanin A.

The isoflavone biochanin A was previously shown to promote weight gain in growing steers by selectively inhibiting rumen bacteria-like growth-promoting feed antibiotics. The hypothesis that biochanin A inhibited the action of drug efflux pumps was tested by enumerating tetracycline-insensitive bacteria from steers in a subacute rumen acidosis (SARA) challenge. Steers ( = 3/group) treatment groups were forage only, SARA control, SARA with monensin (0.

Profiling of cool-season forage arabinoxylans a validated HPAEC-PAD method.

Cool-season pasture grasses contain arabinoxylans (AX) as their major cell wall hemicellulosic polysaccharide. AX structural differences may influence enzymatic degradability, but this relationship has not been fully explored in the AX from the vegetative tissues of cool-season forages, primarily because only limited AX structural characterization has been performed in pasture grasses. Structural profiling of forage AX is a necessary foundation for future work assessing enzymatic degradability and may also be useful for assessing forage quality and suitability for ruminant feed.

Water- and Ethanol-Soluble Carbohydrates of Temperate Grass Pastures: a Review of Factors Affecting Concentration and Composition.

Temperate grasses contain both water- and ethanol-soluble carbohydrates. Water-soluble carbohydrates (WSCs) of temperate grasses include glucose, fructose, sucrose, and fructans (fructose-based polymers) of varying lengths. Ethanol-soluble carbohydrates (ESCs) consist of glucose, fructose, sucrose, and shorter fructans.

Ethanol-Soluble Carbohydrates of Cool-Season Grasses: Prediction of Concentration by Near-Infrared Reflectance Spectroscopy (NIRS) and Evaluation of Effects of Cultivar and Management.

Ethanol-soluble carbohydrates (ESCs) of cool-season grasses include mono- and disaccharides and sometimes short-chain fructans, which may exacerbate the risk of pasture-associated laminitis. A calibration for prediction of ESC concentrations by near-infrared reflectance spectroscopy (NIRS) was developed from 323 samples of four cool-season grass species (orchardgrass, Kentucky bluegrass, tall fescue, and perennial ryegrass) across 10 cultivars collected in central Kentucky in the morning and afternoon over two growing seasons. The calibration, which had accuracy above 95%, was used to predict ESC concentrations of 1,532 samples from the second growing season.

Water-Soluble Carbohydrates of Cool-Season Grasses: Prediction of Concentrations by Near-Infrared Reflectance Spectroscopy and Evaluation of Effects of Genetics, Management, and Environment.

An excess of water-soluble carbohydrates (WSCs) can present a risk for horses prone to pasture-associated laminitis or some other metabolic conditions. Determining WSC concentrations in commonly grazed cool-season grasses, at different times of day and under different fertilization treatments, can help optimize grazing times and management strategies. The goals of this study were to develop a near-infrared reflectance spectroscopy (NIRS) calibration curve for WSC and to apply it to four cool-season grass species.

Isoflavone supplementation, via red clover hay, alters the rumen microbial community and promotes weight gain of steers grazing mixed grass pastures.

Biochanin A, an isoflavone present in the pasture legume red clover (Trifloium pratense L.), alters fermentation in the rumen of cattle and other ruminants. Biochanin A inhibits hyper-ammonia-producing bacteria and promotes cellulolytic bacteria and fiber catalysis in vitro and ex vivo.

Seasonal and Diurnal Variation in Water-Soluble Carbohydrate Concentrations of Repeatedly Defoliated Red and White Clovers in Central Kentucky.

Nonstructural carbohydrates of pasture plants, comprising water-soluble carbohydrates (WSCs) and starch, may contribute to excessive consumption of rapidly fermentable carbohydrates by grazing horses. Seasonal and diurnal variation in WSCs were studied in red (Trifolium pratense L.) and white clovers (Trifolium repens L.

Effects of Harvest Date, Sampling Time, and Cultivar on Total Phenolic Concentrations, Water-Soluble Carbohydrate Concentrations, and Phenolic Profiles of Selected Cool-Season Grasses in Central Kentucky.

Grasses are a source of nutrients for grazing horses. However, water-soluble carbohydrates (WSCs) of grasses have been implicated in some equine health issues. Grasses also contain phenolic compounds, whose sensory and antimicrobial properties may affect forage intake by horses and horse health.

Liberation of recalcitrant cell wall sugars from oak barrels into bourbon whiskey during aging.

Oak barrels have been used by humans for thousands of years to store and transport valuable materials. Early settlers of the United States in Kentucky began charring the interior of new white oak barrels prior to aging distillate to create the distinctively flavored spirit we know as bourbon whiskey. Despite the unique flavor and cultural significance of "America's Spirit", little is known about the wood-distillate interaction that shapes bourbon whiskey.

Hops ( L.) Bitter Acids: Modulation of Rumen Fermentation and Potential As an Alternative Growth Promoter.

Antibiotics can improve ruminant growth and efficiency by altering rumen fermentation selective inhibition of microorganisms. However, antibiotic use is increasingly restricted due to concerns about the spread of antibiotic-resistance. Plant-based antimicrobials are alternatives to antibiotics in animal production.

Mitigation of Ergot Vasoconstriction by Clover Isoflavones in Goats (Capra hircus).

Ergot alkaloids produced by a fungal endophyte (Epichloë coenophiala; formerly Neotyphodium coenophialum) that infects tall fescue (Lolium arundinaceum) can induce persistent constriction of the vasculature in ruminants, hindering their capability to thermo-regulate core body temperature. There is evidence that isoflavones produced by legumes can relax the vasculature, which suggests that they could relieve ergot alkaloid-induced vasoconstriction and mitigate the vulnerability to severe heat stress in ruminants that graze tall fescue. To test if isoflavones can relieve alkaloid-induced vasoconstriction, two pen experiments were conducted with rumen-fistulated goats (Capra hircus) to determine with ultrasonograpy if isoflavones can (1) promote vascular compliance by countering alkaloid-induced vasoconstriction and (2) relieve already imposed alkaloid-induced vasoconstriction.

Blackpatch of Clover, Cause of Slobbers Syndrome: A Review of the Disease and the Pathogen, Rhizoctonia leguminicola.

Rhizoctonia leguminicola Gough and Elliott is a widely used name for the causal agent of blackpatch disease of red clover (Trifolium pratense L.). This fungal pathogen produces alkaloids (slaframine and swainsonine) that affect grazing mammals.

Soluble Phenolic Compounds in Different Cultivars of Red Clover and Alfalfa, and their Implication for Protection against Proteolysis and Ammonia Production in Ruminants.

Red clover (Trifolium pratense) contains soluble phenolic compounds with roles in inhibiting proteolysis and ammonia production. Alfalfa (Medicago sativa) has been found to have a low phenolic content, but few alfalfa and red clover cultivars have been compared for phenolic content. Total soluble phenolics were quantified by a Folin-Ciocalteu colorimetric assay in nine red clover and 27 alfalfa cultivars.

Thin-layer chromatographic (TLC) separations and bioassays of plant extracts to identify antimicrobial compounds.

A common screen for plant antimicrobial compounds consists of separating plant extracts by paper or thin-layer chromatography (PC or TLC), exposing the chromatograms to microbial suspensions (e.g. fungi or bacteria in broth or agar), allowing time for the microbes to grow in a humid environment, and visualizing zones with no microbial growth.

Factors affecting the separation and bioactivity of red clover (Trifolium pratense) extracts assayed against Clostridium sticklandii, a ruminal hyper ammonia-producing bacterium.

Red clover (Trifolium pratense) is rich in phenolic compounds. Both the crude phenolic extract and biochanin A, an isoflavonoid component of the extract, suppress growth of Clostridium sticklandii, a bovine, Gram-positive, ruminal hyper-ammonia producing bacterium (HAB). The purpose of this study was to determine if other components of red clover extract contributed to its anti-HAB activity.

Effects of pH, sample size, and solvent partitioning on recovery of soluble phenolic acids and isoflavonoids in leaves and stems of red clover (Trifolium pratense cv. Kenland).

Several extraction parameters were tested to determine optimal conditions for extracting phenolics from leaves and stems of red clover (Trifolium pratense L. cv. Kenland), with the goal of using extracts in bioassays and in assessment of phenolic profiles.

A validated method for gas chromatographic analysis of gamma-aminobutyric acid in tall fescue herbage.

Gamma-aminobutyric acid (GABA) is an inhibitory neurotransmitter in animals that is also found in plants and has been associated with plant responses to stress. A simple and relatively rapid method of GABA separation and quantification was developed from a commercially available kit for serum amino acids (Phenomenex EZ:faast) and validated for tall fescue (Festuca arundinacea). Extraction in ethanol/water (80:20, v/v) at ambient temperature yielded detectable amounts of GABA.

A functional genomics investigation of allelochemical biosynthesis in Sorghum bicolor root hairs.

Sorghum is considered to be one of the more allelopathic crop species, producing phytotoxins such as the potent benzoquinone sorgoleone (2-hydroxy-5-methoxy-3-[(Z,Z)-8',11',14'-pentadecatriene]-p-benzoquinone) and its analogs. Sorgoleone likely accounts for much of the allelopathy of Sorghum spp., typically representing the predominant constituent of Sorghum bicolor root exudates.

Increased alpha-tocopherol content in soybean seed overexpressing the Perilla frutescens gamma-tocopherol methyltransferase gene.

Tocopherols, with antioxidant properties, are synthesized by photosynthetic organisms and play important roles in human and animal nutrition. In soybean, gamma-tocopherol, the biosynthetic precursor to alpha-tocopherol, is the predominant form found in the seed, whereas alpha-tocopherol is the most bioactive component. This suggests that the final step of the alpha-tocopherol biosynthetic pathway catalyzed by gamma-tocopherol methyltransferase (gamma-TMT) is limiting in soybean seed.

Detoxification and transcriptome response in Arabidopsis seedlings exposed to the allelochemical benzoxazolin-2(3H)-one.

Benzoxazolin-2(3H)-one (BOA) is an allelochemical most commonly associated with monocot species, formed from the O-glucoside of 2,4-dihydroxy-2H-1,4-benzoxazin-3(4H)-one by a two-step degradation process. The capacity of Arabidopsis to detoxify exogenously supplied BOA was analyzed by quantification of the major known metabolites BOA-6-OH, BOA-6-O-glucoside, and glucoside carbamate, revealing that detoxification occurs predominantly through O-glucosylation of the intermediate BOA-6-OH, most likely requiring the sequential action of as-yet-unidentified cytochrome P450 and UDP-glucosyltransferase activities. Transcriptional profiling experiments were also performed with Arabidopsis seedlings exposed to BOA concentrations, representing I(50) and I(80) levels based on root elongation inhibition assays.

Chromatographic separation and in vitro activity of sorgoleone congeners from the roots of sorghum bicolor.

Sorgoleone, 2-hydroxy-5-methoxy-3-[(8'Z,11'Z)-8',11',14'-pentadecatriene]-p-benzoquinone (1), and its corresponding hydroquinone are the major components of the root exudate of Sorghum bicolor. The name sorgoleone includes minor analogues differing in the length or degree of unsaturation of the 3-alkyl side chain. These compounds are known to be phytotoxic, probably through inhibition of photosystem II (PSII) driven oxygen evolution, as previously demonstrated for 1.

Elucidation of the biosynthetic pathway of the allelochemical sorgoleone using retrobiosynthetic NMR analysis.

NMR analyses of the labeling pattern obtained using various 13C-labeled precursors indicated that both the lipid tail and the quinone head of sorgoleone, the main allelopathic component of the oily root exudate of Sorghum bicolor, were derived from acetate units, but that the two moieties were synthesized in different subcellular compartments. The 16:3 fatty acid precursor of the tail is synthesized by the combined action of fatty-acid synthase and desaturases most likely in the plastids. It is then exported out of the plastids and converted to 5-pentadecatriene resorcinol by a polyketide synthase.

Cross-talk between the cytosolic mevalonate and the plastidial methylerythritol phosphate pathways in tobacco bright yellow-2 cells.

In plants, two pathways are utilized for the synthesis of isopentenyl diphosphate, the universal precursor for isoprenoid biosynthesis. The key enzyme of the cytoplasmic mevalonic acid (MVA) pathway is 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR). Treatment of Tobacco Bright Yellow-2 (TBY-2) cells by the HMGR-specific inhibitor mevinolin led to growth reduction and induction of apparent HMGR activity, in parallel to an increase in protein representing two HMGR isozymes.