Effects of prepartum metabolizable protein supply and management strategy on lactational performance and blood biomarkers in dairy cows during early lactation.

Our objective was to investigate the effects of prepartum metabolizable protein (MP) supply and management strategy on milk production and blood biomarkers in early lactation dairy cows. Ninety-six multigravida Holstein cows were used in a randomized complete block design study, blocked by calving date, and then assigned randomly to 1 of 3 treatments within block. Cows on the first treatment were fed a far-off lower MP diet [MP = 83 g/kg of dry matter (DM)] between -55 and -22 d before expected calving and then a close-up lower MP diet (MP = 83 g/kg of DM) until parturition (LPLP). Cows on the second treatment were fed the far-off lower MP diet between -55 to -22 d before expected parturition and then a prepartum higher MP diet (MP = 107 g/kg of DM) until calving (LPHP). Cows on the third treatment had a shortened 43-d dry period and were fed the prepartum higher MP diet from dry-off to parturition (SDHP). After calving, cows received the same fresh diet from d 0 to 14 and the same high diet from d 15 to 84. Data were analyzed separately for wk -6 to -1 and wk 1 to 12, relative to parturition. Dry matter intake from wk -6 to -1 was not different between LPHP and LPLP and increased for SDHP compared with LPLP. In contrast, dry matter intake for wk 1 to 12 postpartum did not change for LPHP versus LPLP or for SDHP versus LPLP. Compared with LPLP cows, LPHP cows had lower energy-corrected milk yield and tended to have decreased milk fat yield during wk 1 to 12 of lactation. Conversely, yields of energy-corrected milk and milk fat and protein were similar for SDHP compared with LPLP. Plasma urea N during wk -3 to -1 increased for LPHP versus LPLP and for SDHP versus LPLP; however, no differences in plasma urea N were observed postpartum. Elevated prepartum MP supply did not modify circulating total fatty acids, ß-hydroxybutyrate, total protein, albumin, or aspartate aminotransferase during the prepartum and postpartum periods. Increased MP supply prepartum combined with a shorter dry period (SDHP vs. LPLP) tended to increase whole-blood ß-hydroxybutyrate postpartum; however, other blood metabolites were not affected. Taken together, under the conditions of this study, elevated MP supply in close-up diets reduced milk production without affecting blood metabolites in multiparous dairy cows during early lactation. A combination of a shorter dry period and increased prepartum MP supply (i.e., SDHP vs. LPLP) improved prepartum dry matter intake without modifying energy-corrected milk yield and blood biomarkers in early lactation cows.

Reducing metabolizable protein supply: Effects on milk production, blood metabolites, and health in early-lactation dairy cows.

Our objective was to evaluate the effect of metabolizable protein (MP) supply on milk production, blood metabolites, and health in dairy cows during early lactation. Three experimental diets were formulated to contain 114, 107, 101 g of MP/kg of dry matter (DM; 114MP, 107MP, and 101MP, respectively) with crude protein contents of 17.0, 16.2, and 15.3% of DM, respectively. One hundred multiparous Holstein cows were fed 1 of these 3 diets during wk 1 to 3 and wk 4 to 13 of lactation in one of the following sequences: (1) 114MP and 107MP (114MP/107MP), (2) 114MP and 101MP (114MP/101MP), or (3) 101MP and 101MP (101MP/101MP). During wk 1 to 3, the 114MP and 101MP treatments were 20 and 27% deficient in estimated MP, respectively. From wk 4 to 13, the 114MP/107MP, 114MP/101MP, and 101MP/101MP treatments were 8, 12, and 13% deficient in estimated MP, respectively. Data were analyzed separately for wk 1 to 3, 4 to 13, and 1 to 13. Dry matter intake and energy-corrected milk (ECM) yield were not affected by treatment during wk 4 to 13 or wk 1 to 13; however, ECM yield decreased for 101MP versus 114MP from wk 1 to 3. Similarly, feed efficiency was not affected by treatment from wk 4 to 13 or wk 1 to 13, and was reduced with 101MP versus 114MP during wk 1 to 3. Milk N efficiency tended to increase for 101MP versus 114MP for wk 1 to 3 and increased with 101MP/101MP and 114MP/101MP relative to 114MP/107MP during wk 4 to 13 and wk 1 to 13. Treatment had no influence on yields and concentrations of milk components from wk 4 to 13 or wk 1 to 13; however, compared with 114MP, feeding 101MP tended to decrease milk fat yield and decreased yields of milk true protein and lactose for wk 1 to 3. Both milk and blood urea N concentrations decreased for 101MP/101MP and 114MP/101MP relative to 114MP/107MP during wk 4 to 13 and wk 1 to 13, and were reduced with feeding 101MP versus 114MP from wk 1 to 3. Treatment had no effect on the incidence of diseases in cows throughout the study. Serum concentrations of total fatty acids, albumin, and aspartate aminotransferase did not differ between 101MP and 114MP; however, serum ß-hydroxybutyrate concentration was lower in cows receiving 101MP during the first 3 wk of lactation. Compared with 114MP, feeding 101MP during wk 1 to 3 increased plasma concentrations of creatinine and 3-methylhistidine (3-MHis) but did not change the ratio of plasma 3-MHis to creatinine. We found no differences in plasma creatinine or the ratio of 3-MHis-to-creatinine among treatments from wk 4 to 13; however, 101MP/101MP and 114MP/101MP had elevated plasma 3-MHis compared with 114MP/107MP. Treatment had no effect on body weight and body condition score over the duration of the study. Collectively, despite reduced milk production for the first 3 wk of lactation, feeding the 101MP/101MP treatment sustained lactational performance and improved milk N efficiency without negatively affecting the frequency of diseases in dairy cows during the first 13 wk postpartum.

Effect of stocking density and effective fiber on the ruminal bacterial communities in lactating Holstein cows.

Overstocking can be a major issue in the dairy cattle industry, leading to negative changes in feeding and resting behavior. Additional stress imposed and alterations in feeding behavior may significantly impact the rumen microbiome. The rumen microbiome is responsible for the successful conversion of feed to usable energy for its host. Thus, understanding the effects of stocking density on the rumen microbiome is imperative for further elucidation of potentially negative consequences of overstocking in dairy cattle. This study implemented a Latin Square design accounting for four pens of cattle and four treatment periods so that all treatment combinations were assigned to every pen during one period of the study. Two treatment factors, including two levels of physically effective neutral detergent fiber, achieved with addition of chopped straw, and stocking density (100% vs. 142%) of freestalls and headlocks, were combined and tested within a factorial treatment design. Within each pen, three or four cannulated cows (n = 15 total) were sampled for rumen content on the final day of each treatment period. Each treatment was randomly assigned to a single pen for a 14-day period. The V1-V3 hypervariable regions of the 16S rRNA gene were targeted for bacterial analyses. Variables with approximately normally-distributed residuals and a Shapiro-Wilk statistic of ≥0.85 were analyzed using a mixed model analysis of variance with the GLIMMIX procedure with fixed effects of feed (straw vs. no straw), stocking density (100% vs. 142%), and the interaction of feed × stocking density, and random effects of pen, period, feed × stocking × pen × period. Pen was included as the experimental unit in a given period and the sampling unit as cow. Variables included Shannon's Diversity Index, Faith's phylogenetic diversity index, chao1, observed OTU, and Simpson's evenness E as well as most individual taxa. Data were analyzed in SAS 9.4 utilizing the GLIMMIX procedure to perform mixed model analysis of variance. If data were not normally distributed, a ranked analysis was performed. No differences were observed in α-diversity metrics by fiber or stocking density (P > 0.05). Beta diversity was assessed using weighted and unweighted Unifrac distances in QIIME 1.9.1 and analyzed using ANOSIM. No differences were observed in weighted (P = 0.6660; R = -0.0121) nor unweighted (P = 0.9190; R = -0.0261) metrics and R values suggested similar bacterial communities among treatments. At the phylum level, Tenericutes differed among treatments with an interaction of stocking density by feed (P = 0.0066). At the genus level, several differences were observed by treatment, including Atopobium (P = 0.0129), unidentified members of order RF39 (P = 0.0139), and unidentified members of family Succinivibrionaceae (P = 0.0480). Although no diversity differences were observed, taxa differences may indicate that specific taxa are affected by the treatments, which may, in turn, affect animal production.

Nutrition-induced ketosis alters metabolic and signaling gene networks in liver of periparturient dairy cows.

Dairy cows are highly susceptible after parturition to developing liver lipidosis and ketosis, which are costly diseases to farmers. A bovine microarray platform consisting of 13,257-annotated oligonucleotides was used to study hepatic gene networks underlying nutrition-induced ketosis. On day 5 postpartum, 14 Holstein cows were randomly assigned to ketosis-induction (n = 7) or control (n = 7) groups. Cows in the ketosis-induction group were fed at 50% of day 4 intake until they developed signs of clinical ketosis, and cows in the control group were fed ad libitum throughout the treatment period. Liver was biopsied at 10-14 (ketosis) or 14 days postpartum (controls). Feed restriction increased blood concentrations of nonesterified fatty acids and beta-hydroxybutyrate, but decreased glucose. Liver triacylglycerol concentration also increased. A total of 2,415 genes were altered by ketosis (false discovery rate = 0.05). Ingenuity Pathway Analysis revealed downregulation of genes associated with oxidative phosphorylation, protein ubiquitination, and ubiquinone biosynthesis with ketosis. Other molecular adaptations included upregulation of genes and nuclear receptors associated with cytokine signaling, fatty acid uptake/transport, and fatty acid oxidation. Genes downregulated during ketosis included several associated with cholesterol metabolism, growth hormone signaling, proton transport, and fatty acid desaturation. Feed restriction and ketosis resulted in previously unrecognized alterations in gene network expression underlying key cellular functions and discrete metabolic events. These responses might help explain well-documented physiological adaptations to reduced feed intake in early postpartum cows and, thus, provide molecular targets that might be useful in prevention and treatment of liver lipidosis and ketosis.

Plane of nutrition prepartum alters hepatic gene expression and function in dairy cows as assessed by longitudinal transcript and metabolic profiling.

Liver metabolism and health in dairy cows during the periparturient period are affected by plane of nutrition prepartum. Long-term adaptations in hepatic gene expression are important for complete understanding of liver function. We examined temporal gene expression profiles during the dry period and early lactation in liver of Holstein cows fed moderate dietary energy ad libitum or restricted during the entire dry period using a microarray consisting of 7,872 annotated cattle cDNA inserts and quantitative RT-PCR. We identified 85 genes with expression patterns that were affected by level of energy intake prepartum over time. Restricted energy intake prepartum resulted in more pronounced upregulation of genes with key functions in hepatic fatty acid oxidation (CPT1A, ADIPOR2), gluconeogenesis (PC), and cholesterol synthesis (SC4MOL). Ad libitum feeding upregulated a number of genes associated with liver triacylglycerol synthesis (DGAT1) and proinflammatory cytokines (TNFAIP3). Genomic responses to ad libitum feeding were accompanied by increased incorporation of palmitate to esterified products in vitro and increased liver triacylglycerol concentration in vivo. Overall, gene expression profiles due to plane of nutrition prepartum partly explained differences in rates of liver palmitate metabolism, blood serum metabolite concentrations, and liver tissue triacylglycerol concentration. Our data show that moderate overfeeding of energy in the dry period, in the absence of obesity, results in transcriptional changes predisposing cows to fatty liver and perhaps compromising overall liver health during the periparturient period. In this context, controlled energy intake may confer an advantage to the cow by triggering hepatic molecular adaptations well ahead of parturition.

Temporal gene expression profiling of liver from periparturient dairy cows reveals complex adaptive mechanisms in hepatic function.

Long-term molecular adaptations in liver from high-producing dairy cows are virtually unknown. Liver from five Holstein cows was biopsied at -65, -30, -14, +1, +14, +28, and +49 days relative to parturition for transcript profiling using a microarray consisting of 7,872 annotated cattle cDNA inserts. More than 5,000 cDNA elements represented on the microarray were expressed in liver. From this set we identified 62 differentially expressed genes related to physiological state, with a false discovery rate threshold of P = 0.20. The dominant expression pattern consisted of upregulation from day -30 through day +1, followed by downregulation through day +28. There was a threefold decrease from day -65 through day +14 in expression of IGFBP3, GSTM5, and PDPK1. These genes mediate IGF-I transport, oxidative stress, and glucose homeostasis, respectively. IGFBP3, EIF4B, and GSTM5 mRNA levels were positively correlated with blood serum total protein. Correlation analysis showed positive associations between serum nonesterified fatty acids and mRNA expression for SAA1, CPT1A, ACADVL, and TFAP2A. Transcript levels of ACSL1, PPARA, and TFAP2A were positively correlated with serum beta-hydroxybutyrate. Expression patterns for certain genes (e.g., IGFBP3, HNF4A, GPAM) revealed adaptations commencing well ahead of parturition, suggesting they are regulated by factors other than periparturient hormonal environment. Results provide evidence that hepatic inflammatory responses occurring near parturition initiate or augment adipose catabolism. In this context, cytokines, acute-phase proteins, and serum nonesterified fatty acids are key players in periparturient cow metabolism. We propose a model for integrating gene expression, metabolite, and liver composition data to explain physiological events in placenta, adipose, and liver during the periparturient period.