Protein is an important fraction within bovine milk. This milk protein is not only vital for calf growth and development, but also includes bioactive proteins and peptides that have been shown to enhance the health of animals and humans. Research efforts are focusing on factors, such as nutrition, that can influence the quantity and profile of proteins within the bovine milk proteome. The research outlined herein investigated the impact of diet on the bovine milk proteome. The first experiment examined whether dietary inclusion of grape marc (GM), a condensed tannin (CT) containing by-product from the viticulture industry, could alter the bovine milk proteome through altered nitrogen (N) metabolism. In this experiment, 10 lactating Holstein cows were fed either 2.0 kg dry matter (DM)/ cow/ day of beet pulp: soy hulls in a 50% mixture (control), or 1.5 kg DM/ cow/ day of GM as part of a balanced dairy cow ration for a 28-d trial. Milk samples were obtained for analysis of the high- and low-abundance protein fractions. Skimmed milk samples collected for high-abundance protein analysis were measured using high performance liquid chromatography (HPLC), and liquid-chromatography tandem mass spectrometry (LC-MS/MS) was used to identify proteins in the low-abundance protein enriched fraction. Skimmed milk samples collected for low-abundance milk protein analysis were fractionated and enriched to remove higher abundance proteins. Enriched milk samples were then digested and labeled with isobaric tandem mass tags (TMT) prior to protein identification using LC-MS/MS analysis. There were no changes in the high-abundance protein fraction in response to diet; however, 16 of 127 low-abundance proteins were identified at different relative-abundances due to diet (P ≤ 0.05). While there were no alterations in the metabolic or N status of animals due to GM supplementation, the 12% change in the low-abundance milk protein fraction highlighted the potential for dietary alteration of the bovine milk proteome. A second experiment evaluated the inclusion of alternative forage crops (AFC) as a means to alter the bovine milk proteome. In this experiment, both the skimmed milk and milk fat globule membrane (MFGM) protein fractions were included in analysis. Milk samples were collected from 16 lactating Jersey cattle included in a 21-d grazing experiment, where cows were offered one of two diets. The control group (CON, n=8) grazed a grass-legume pasture mixture containing orchardgrass (Dactylis glomerata), timothy (Phleum pratense), Kentucky bluegrass (Poa pratensis), and white clover (Trifolium repens). The treatment group (AFC, n=8) grazed a similar base pasture that was strip-tilled with oat (Avena sativa), buckwheat (Fagopyrum esculentum), and chickling vetch (Lathyrus sativus) so that the AFC species comprised 10% of the AFC group’s pasture DM intake (DMI). Milk samples were collected for HPLC analysis of the high abundance milk proteins, and LC-MS/MS analysis of the low abundance protein enriched skim milk fraction and MFGM-associated protein fraction. Cows that grazed pastures containing AFC had higher αs1-CAS content (P = 0.005), and higher relative-abundances of 7 low-abundance proteins within the skim milk and MFGM fractions (P ≤ 0.05). While it is plausible that the inclusion of AFC in pasture increased nutrient availability to the mammary gland, the specific mechanisms that could have caused the shifts observed remain unclear. Further investigation is necessary to fully understand the role of diet and the milk protein profile.

This experiment examined the forage potential of hemp (Cannabis sativa) and kenaf (Hibiscus cannabinus). The objectives were to evaluate yield and forage nutritive value (i.e. NDF, ADF, ADL, IVTD, and CP) fluctuations over the course of a growing season based on planting date, morphological composition, and management. Three types of hemp (grain, fiber, and a dual- purpose type) and kenaf were planted on two dates and were sampled approximately every two weeks throughout the growing season at the University of Kentucky (UK) Research Farm in Lexington, KY. Subsamples were separated into morphological components (i.e. leaf, flowers, stem, core fiber, and bast fiber) while the remainder of the sample was ground for laboratory analysis. All samples were scanned in Foss 6500 NIRS and wet chemistry analytical methods were utilized on a subset of samples to develop equations to predict the nutritive value of the remaining samples. Significant interactions for forage type, planting date, and harvest time were observed for yield, % floral components, % bast, and ADL. Significant interactions occurred between planting date and harvest date as well as type and harvest date for NDF, ADF, digestibility, crude protein, % leaf, % core, and % stem. Overall, forage nutritive value declined with increased plant maturity. The later planting date reduced the vegetative growth period, resulting in reduced leaf content, yield, and forage nutritive value. The performance of kenaf in this study indicates that it may be a better alternative forage than hemp due to remaining vegetative longer and having superior nutritive value. Better selection and the development of new hemp varieties with different photoperiod requirements could lengthen the vegetative state and may result in yields and nutritive values that are more competitive with kenaf and other typical forages.

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