Influence of prepartum dietary cation-anion difference and the magnitude of calcium decline at the onset of lactation on mineral metabolism and physiological responses.

The onset of lactation is characterized by substantially altered calcium (Ca) metabolism; recently, emphasis has been placed on understanding the dynamics of blood Ca in the peripartal cow in response to this change. Thus, the aim of our study was to delineate how prepartum dietary cation-anion difference (DCAD) diets and the magnitude of Ca decline at the onset of lactation altered blood Ca dynamics in the periparturient cow. Thirty-two multiparous Holstein cows were blocked by parity, previous 305-d milk yield and expected parturition date, and randomly allocated to either a positive (+120 mEq/kg) or negative (-120 mEq/kg) DCAD diet from 251 d of gestation until parturition (n = 16/diet). Immediately after parturition cows were continuously infused for 24 h with (1) an intravenous solution of 10% dextrose or (2) Ca gluconate (CaGlc) to maintain blood ionized (iCa) concentrations at ∼1.2 mM (normocalcemia) to form 4 treatment groups (n = 8/treatment). Blood was sampled every 6 h from 102 h before parturition until 96 h after parturition and every 30 min during 24 h continuous infusion. Cows fed a negative DCAD diet prepartum exhibited a less pronounced decline in blood iCa approaching parturition with lesser magnitude of decline relative to positive DCAD-fed cows. Cows fed a negative DCAD diet prepartum required lower rates of CaGlc infusion to maintain normocalcemia in the 24 h postpartum relative to positive DCAD-fed cows. Infusion of CaGlc disrupted blood Ca and P dynamics in the immediate 24 h after parturition and in the days following infusion. Collectively, these data demonstrate that prepartum negative DCAD diets facilitate a more transient hypocalcemia and improve blood Ca profiles at the onset of lactation whereas CaGlc infusion disrupts mineral metabolism.

Income is not an equalizer: health development inequities by ethnoracial backgrounds in California kindergartners.

BACKGROUND: Early childhood health development is positively associated with income, but the strength of this relationship with ethnoracial background remains unclear. This study examined the extent of health development inequities among California kindergarteners based on ethnoracial backgrounds and neighborhood-level income. METHODS: This cross-sectional study assessed health development inequities by analyzing neighborhood-level income, ethnoracial background, and health development data for California kindergarteners. Student-level data (n = 106,574) were collected through teacher report between 2010-2020 across 52 school districts and 964 schools. Student addresses were geocoded and linked to American Community Survey neighborhood income levels. Health development was measured using the Early Development Instrument, a population-level measure which includes physical health and well-being, social competence, emotional maturity, language and cognitive development, and communication skills and general knowledge domains. Outcomes included being "on-track" in each domain as well as overall health development. RESULTS: Using a Generalized Estimation Equation with a log-link function, while accounting for interactions between ethnoracial background, income, and income-squared, we found significant health development inequities by ethnoracial background and neighborhood-level income. Regarding overall health development, as well as the physical, social and emotional domains, Black students had a lower likelihood of being on-track compared to the weighted average across income levels, whereas Asian students surpassed the weighted average. White students exhibited the steepest slope, and at the lowest income levels, their health development scores were akin to their Black and Hispanic/Latino/a low-income counterparts but resembled their Asian counterparts at higher income levels. For the general knowledge and communication domain, white students consistently had the highest likelihood of being on-track, while Hispanic/Latino/a students had the lowest likelihood across all income levels. CONCLUSION: This study examines health development inequities among California kindergarteners in diverse communities. Our analysis shows that the relationship between neighborhood-level income and kindergartners' health development varies by domain and is weaker for students of color. Given the scarcity of population-level data on health development outcomes, these analyses offer valuable insights for identifying ecosystems necessitating support in promoting equitable early childhood health development.

Supplementing pre- and probiotic ingredients to feedlot steers: effects on health, growth performance, and physiological responses.

Feedlot diets are often enriched with additives to mitigate health disorders and promote cattle performance, including the feed-grade antimicrobials monensin and tylosin. However, alternative feeding strategies are warranted given the increasing regulations regarding the use of antimicrobials in feedlot diets. This study evaluated the performance, physiological, and health responses of feedlot cattle offered a synbiotic supplement (yeast-derived prebiotic + Bacillus subtilis probiotic), which replaced or was fed in conjunction with monensin and tylosin. Angus-influenced steers (n = 192) from four different cowherds were weaned on day -1 and transported (800 km) to the feedlot. Steers were allocated to 1 of 24 pens (eight steers/pen) upon arrival on day 0. Pens were assigned to receive (n = 8/treatment) a total-mixed ration (TMR) containing: (1) monensin and tylosin (RT; 360 mg/steer daily from Rumensin and 90 mg/steer daily from Tylan; Elanco Animal Health, Greenfield, IN, USA), (2) yeast-derived ingredient and B. subtilis probiotic (CC; 18 g/steer daily of Celmanax and 28 g/steer daily of Certillus; Church and Dwight Co., Inc., Princeton, NJ, USA), or (3) a combination of RT and CC (RTCC). Steers were slaughtered according to BW in four groups balanced by treatment and pens and received treatments for 252 ± 4 days. No treatment effects were detected (P ≥ 0.17) for steer BW gain and morbidity responses. Mean TMR intake was greater and gain:feed ratio was less (P ≤ 0.01) in CC compared with RT and RTCC steers. Mean plasma leptin concentration was greater (P ≤ 0.05) in CC compared with RT and RTCC steers. Steers receiving CC had greater (P ≤ 0.04) concentrations of plasma cortisol, haptoglobin, glucose, and beta-hydroxybutyrate, and less (P ≤ 0.05) concentration of non-esterified fatty acids compared with RT and RTCC steers on day 14 of the experiment. Carcass marbling was greater (P = 0.01) in CC compared with RT steers and tended to be greater (P = 0.07) in RTCC compared with RT steers. Proportion of carcasses that graded Choice or better and Longissimus muscle area were greater (P ≤ 0.05) in CC and RTCC compared with RT steers. Incidence of liver abscesses was less (P = 0.01) in RTCC compared with CC steers and tended to be less (P = 0.09) in RT compared with CC steers. Results from this experiment indicate that the synbiotic supplement may replace monensin and tylosin without reducing steer BW gain, with potential improvements to carcass quality traits.

Characterizing ruminal acidosis risk: A multiherd, multicountry study.

A multicenter observational study was conducted on early lactation Holstein cows (n = 261) from 32 herds from 3 regions (Australia, AU; California, CA; and Canada, CAN) to characterize their risk of acidosis into 3 groups (high, medium, or low) using a discriminant analysis model previously developed. Diets ranged from pasture supplemented with concentrates to total mixed ration (nonfiber carbohydrates = 17 to 47 and neutral detergent fiber = 27 to 58% of dry matter). Rumen fluid samples were collected <3 h after feeding and analyzed for pH, and ammonia, d- and l-lactate, and volatile fatty acid (VFA) concentrations. Eigenvectors were produced using cluster and discriminant analysis from a combination of rumen pH, and ammonia, d-lactate, and individual VFA concentrations and were used to calculate the probability of the risk of ruminal acidosis based on proximity to the centroid of 3 clusters. Bacterial 16S ribosomal DNA sequence data were analyzed to characterize bacteria. Individual cow milk volume, fat, protein, and somatic cell count values were obtained from the closest herd test to the rumen sampling date (median = 1 d before rumen sampling). Mixed model analyses were performed on the markers of rumen fermentation, production characteristics, and the probability of acidosis. A total of 26.1% of the cows were classified as high risk for acidosis, 26.8% as medium risk, and 47.1% as low risk. Acidosis risk differed among regions with AU (37.2%) and CA (39.2%) having similar prevalence of high-risk cows and CAN only 5.2%. The high-risk group had rumen phyla, fermentation, and production characteristics consistent with a model of acidosis that reflected a rapid rate of carbohydrate fermentation. Namely, acetate to propionate ratio (1.98 ± 0.11), concentrations of valerate (2.93 ± 0.14 mM), milk fat to protein ratio (1.11 ± 0.047), and a positive association with abundance of phylum Firmicutes. The medium-risk group contains cows that may be inappetant or that had not eaten recently or were in recovery from acidosis. The low-risk group may represent cattle that are well fed with a stable rumen and a slower rumen fermentation of carbohydrates. The high risk for acidosis group had lower diversity of bacteria than the other groups, whereas CAN had a greater diversity than AU and CA. Rumen fermentation profile, abundance of ruminal bacterial phyla, and production characteristics of early lactation dairy cattle from 3 regions were successfully categorized in 3 different acidosis risk states, with characteristics differing between acidosis risk groups. The prevalence of acidosis risk also differed between regions.

Associations among the genome, rumen metabolome, ruminal bacteria, and milk production in early-lactation Holsteins.

A multicenter observational study to evaluate genome-wide association was conducted in early-lactation Holstein cows (n = 293) from 36 herds in Canada, the USA, and Australia. Phenotypic observations included rumen metabolome, acidosis risk, ruminal bacterial taxa, and milk composition and yield measures. Diets ranged from pasture supplemented with concentrates to total mixed rations (nonfiber carbohydrates = 17 to 47, and neutral detergent fiber = 27 to 58% of dry matter). Rumen samples were collected <3 h after feeding and analyzed for pH, ammonia, d- and l-lactate, volatile fatty acid (VFA) concentrations, and abundance of bacterial phyla and families. Eigenvectors were produced using cluster and discriminant analyses from a combination of pH and ammonia, d-lactate, and VFA concentrations, and were used to estimate the probability of the risk of ruminal acidosis based on proximity to the centroid of 3 clusters, termed high (24.0% of cows), medium (24.2%), and low risk (51.8%) for acidosis. DNA of sufficient quality was successfully extracted from whole blood (218 cows) or hair (65 cows) collected simultaneously with the rumen samples and sequenced using the Geneseek Genomic Profiler Bovine 150K Illumina SNPchip. Genome-wide association used an additive model and linear regression with principal component analysis (PCA) population stratification and a Bonferroni correction for multiple comparisons. Population structure was visualized using PCA plots. Single genomic markers were associated with milk protein percent and the center logged ratio abundance of the phyla Chloroflexi, SR1, and Spirochaetes, and tended to be associated with milk fat yield, rumen acetate, butyrate, and isovalerate concentrations and with the probability of being in the low-risk acidosis group. More than one genomic marker was associated or tended to be associated with rumen isobutyrate and caproate concentrations, and the center log ratio of the phyla Bacteroidetes and Firmicutes and center log ratio of the families Prevotellaceae, BS11, S24-7, Acidaminococcaceae, Carnobacteriaceae, Lactobacillaceae, Leuconostocaceae, and Streptococcaceae. The provisional NTN4 gene, involved in several functions, had pleiotropy with 10 bacterial families, the phyla Bacteroidetes and Firmicutes, and butyrate. The ATP2CA1 gene, involved in the ATPase secretory pathway for Ca2+ transport, overlapped for the families Prevotellaceae, S24-7, and Streptococcaceae, the phylum Bacteroidetes, and isobutyrate. No genomic markers were associated with milk yield, fat percentage, protein yield, total solids, energy-corrected milk, somatic cell count, rumen pH, ammonia, propionate, valerate, total VFA, and d-, l-, or total lactate concentrations, or probability of being in the high- or medium-risk acidosis groups. Genome-wide associations with the rumen metabolome, microbial taxa, and milk composition were present across a wide geographical and management range of herds, suggesting the existence of markers for the rumen environment but not for acidosis susceptibility. The variation in pathogenesis of ruminal acidosis in the small population of cattle in the high risk for acidosis group and the dynamic nature of the rumen as cows cycle through a bout of acidosis may have precluded the identification of markers for acidosis susceptibility. Despite a limited sample size, this study provides evidence of interactions between the mammalian genome, the rumen metabolome, ruminal bacteria, and milk protein percentage.

Performance, health, and physiological responses of newly received feedlot cattle supplemented with pre- and probiotic ingredients.

Nutritional strategies that optimize immunity of feedlot cattle are warranted due to increasing regulations with the use of feed-grade antimicrobials. This study evaluated physiological, health, and performance responses of cattle receiving a synbiotic supplement (yeast-derived prebiotic + Bacillus subtilis probiotic), which replaced feed-grade antimicrobials or were fed in conjunction with monensin during the initial 45 days in the feedlot. Angus-influenced steers (n = 256) were acquired from an auction facility on day -2, and transported (800 km) to the feedlot. Shrunk BW was recorded upon arrival (day -1). Steers were allocated to 1 of 18 pens (day 0), and pens were assigned to receive (n = 6/treatment) a free-choice diet containing: (1) monensin and tylosin (RT; 360 mg/steer daily from Rumensin and 90 mg/steer daily from Tylan; Elanco Animal Health, Greenfield, IN, USA), (2) yeast-derived ingredient and B. subtilis probiotic (CC; 18 g/steer daily of Celmanax and 28 g/steer daily of Certillus; Church and Dwight Co., Inc., Princeton, NJ, USA), or (3) monensin in addition to yeast-derived and B. subtilis ingredients (RCC) as in RT and CC. Steers were assessed for bovine respiratory disease (BRD) and DMI daily. Steer BW was recorded on days 45 and 46, and averaged for final BW. Blood samples were collected on days 0, 7, 17, 31, and 45. Feed intake was greater (P ≤ 0.05) in CC vs. RCC and RT during the initial 3 weeks upon feedlot arrival. No treatment differences were noted (P ≥ 0.41) for average daily gain, BW, and feed efficiency. Incidence of BRD did not differ (P = 0.77) between treatments (average 80.1%). A greater proportion (P ≤ 0.03) of RT steers diagnosed with BRD required a second antimicrobial treatment compared with CC and RCC (57.3, 37.3, and 38.6%, respectively). Removal of steers from the trial due to severe morbidity + mortality was greater (P = 0.02) in RT vs. CC (22.4 and 7.0%), and did not differ (P ≥ 0.16) among RCC (12.9%) vs. RT and CC. Plasma glucose concentrations were greater (P ≤ 0.02) in CC vs. RCC and RT on day 7. Plasma concentrations of nonesterified fatty acids were greater (P ≤ 0.02) in RT and RCC vs. CC on day 7, and in RT vs. CC on day 17. Steers receiving the synbiotic supplement had improved response to BRD treatment, suggesting heightened immunocompetence from partially enhanced metabolism and the nutraceutical effects of B. subtilis and yeast compounds.

Effects of mannan-oligosaccharide and Bacillus subtilis supplementation to preweaning Holstein dairy heifers on body weight gain, diarrhea, and shedding of fecal pathogens.

The objective of this clinical trial was to evaluate the effectiveness of probiotic, prebiotic, and synbiotic supplementation on average daily weight gain (ADG), duration of diarrhea, age at incidence of diarrhea, fecal shedding of Cryptosporidium oocysts, enteric pathogens, and the odds of pneumonia in preweaning dairy heifer calves on a commercial dairy. Feeding prebiotics and probiotics may improve health and production of calves. Hence, healthy Holstein heifer calves (n = 1,801) from a large California dairy were enrolled at 4 to 12 h of age and remained in this study until weaning at 60 d of age. Calves were block randomized to 1 of 4 treatments: (1) control, (2) yeast culture enriched with mannan-oligosaccharide (prebiotic), (3) Bacillus subtilis (probiotic), and (4) combination of both products (synbiotic), which were fed in milk twice daily from enrollment until weaning. Serum total protein at enrollment and body weight at 7, 42, and 56 d of age were measured. Fecal consistency was assessed daily for the entire preweaning period. A subgroup of 200 calves had fecal samples collected at 7, 14, 21, and 42 d for microbial culture and enumeration of Cryptosporidium oocysts by direct fluorescent antibody staining. Synbiotic-treated calves had 19 g increased ADG compared with control calves for overall ADG, from 7 to 56 d. From 42 to 56 d, prebiotic-treated calves had 85 g greater ADG and synbiotic-treated calves had 78 g greater ADG than control calves. There was no difference in duration of the first diarrhea episode, hazard of diarrhea, or odds of pneumonia per calf with treatment. Probiotic-treated calves had 100 times lower fecal shedding of Cryptosporidium oocysts at 14 d and prebiotic-treated calves had fewer Escherichia coli and pathogenic E. coli at 42 d compared with control calves. Although there were no effects on duration of diarrhea or pneumonia incidence, greater ADG in the late preweaning period may reflect treatment effects on enteric pathogens during the rearing process. The decreased shedding of Cryptosporidium should reduce infectious pressure, environmental contamination, and public health risks from Cryptosporidium. Our findings suggest ADG and potential health benefits for calves fed prebiotics, probiotics, and synbiotics and can help the dairy industry make informed decisions on the use of these products in dairy production.

Duration and degree of diet-induced metabolic acidosis prepartum alter tissue responses to insulin in dairy cows.

The objective of this experiment was to determine the effects of altering the dietary cation-anion difference (DCAD) fed for the last 21 or 42 d of gestation on glucose metabolism and tissue insulin responsiveness. Ninety parous Holstein cows at 232 d of gestation were assigned randomly to dietary treatments with 2 levels of DCAD (-70 or -180 mEq/kg) fed for 2 durations (short: the last 21 d of gestation; long: the last 42 d of gestation). For the short treatments, a diet with +110 mEq/kg was fed from 232 to 254 d of gestation. Intravenous glucose tolerance tests (IVGTT) were performed at either 250 or 270 d of gestation by infusing 0.25 g of dextrose/kg of body weight within 1 min. The following day, cows underwent an insulin challenge (IC) and received 0.1 IU of insulin/kg of body weight intravenously. Blood was sampled at min -15, -5, and 0 to establish a baseline and from 5 to 180 min relative to infusions; plasma concentrations of glucose, insulin, and fatty acids were determined, and the respective areas under the curves (AUC) were calculated. Liver was sampled after the IVGTT, and adipose tissue was sampled after the IVGTT and IC for quantification of mRNA expression and protein abundance. Reducing the DCAD altered acid-base balance compatible with a compensated metabolic acidosis. At 250 d, reducing the DCAD increased the AUC for glucose and reduced that of insulin following the IVGTT, whereas during the IC, clearance rate decreased and time to half-life of insulin increased with reducing DCAD, resulting in a tendency to a larger AUC for fatty acids. At 270 d, quantitative insulin sensitivity check index and the revised quantitative insulin sensitivity check index were smaller in cows fed the acidogenic diets for the last 42 d of gestation compared with the last 21 d of gestation, thereby suggesting reduced insulin sensitivity. In addition, cows fed for the long duration tended to have greater AUC for glucose but smaller AUC for insulin following an IVGTT than those fed for the short duration, thereby suggesting reduced insulin release and glucose disposal. Treatments did not affect hepatic mRNA expression of G6PC, PCK1, PCK2, and PC or adipose tissue mRNA expression of ATGL, ACC, B2AR, HSL, and PLIN1. On the other hand, for proteins, reducing the DCAD linearly reduced abundance of rabbit anti-mouse protein kinase B (AKT) and tended to reduce rabbit anti-human phosphorylated (Ser-9) glycogen synthase kinase-3 ß (pGSK) and the pGSK:rabbit anti-human glycogen synthase kinase-3 ß (GSK) ratio in hepatic tissue, whereas a linear increase in rabbit anti-human hormone-sensitive lipase (HSL) and rabbit anti-mouse phosphorylated (Ser-660) hormone-sensitive lipase (pHSL) in adipose tissue was observed after the IVGTT at 250 d. Moreover, reducing the DCAD resulted in a linear reduction of AKT and tended to reduce rabbit anti-human acetyl-CoA carboxylase (ACC) but increased pHSL linearly in adipose tissue after an IC at 250 d. Cows fed acidogenic diets for a short duration tended to have less pHSL in adipose tissue than those fed for a long duration after an IVGTT at 270 d. Associations were observed between blood pH and mRNA and protein abundance in hepatic and adipose tissues. Diet-induced metabolic acidosis altered insulin release and insulin signaling, resulting in a shift in adipose tissue metabolism that would favor lipolysis over lipogenesis.

Comparison of milk production, intake, and total-tract nutrient digestion in lactating dairy cattle fed diets containing either wheat middlings and urea, commercial fermentation by-product, or rumen-protected soybean meal.

The objective of this study was to evaluate the effects of a commercially available fermentation by-product in a diet containing adequate rumen-degradable protein (RDP) on milk performance, intake, and total-tract nutrient digestion in lactating dairy cattle. Primiparous (n = 48) and multiparous (n = 144) lactating dairy cattle were stratified by milk production and randomly allocated into 12 pens containing 4 primiparous and 12 multiparous animals each. Cattle averaged 118 d in milk and 712 kg of body weight at trial start. Treatment diets, on a dry matter (DM) basis, consisted of 42% corn silage, 13% alfalfa hay and silage, 20% grain corn, and 25% protein premix containing either soybean meal, wheat middlings, and urea (SBM+U), soybean meal and fermentation by-product (SBM+F), or soybean meal and rumen-protected soybean meal (RP-SBM). All 3 diets provided a similar level (DM basis) of neutral detergent fiber analyzed using α-amylase and sodium sulfite and corrected for ash content (31%), crude protein (CP, 14.9%), starch (26%), and metabolizable energy (2.7 Mcal/kg), and differed in sources of RDP. The trial consisted of a 2-wk adaptation and covariate period during which all cows were fed the RP-SBM diet and covariate measurements were taken. Pens were then randomly allocated to treatments, and weekly measurements of milk production, intake, body weight, and condition score were taken for 10 wk. All data were analyzed using the Proc Mixed procedure in SAS (SAS Institute Inc., Cary, NC). Increased DM intake was observed for cows fed SBM+F compared with cows fed SBM+U and RP-SBM (28.3 vs. 26.9 and 26.7 kg/d, respectively). Cows fed SBM+F produced more energy-corrected milk (45.3 kg/d) compared with cows fed SBM+U and RP-SBM (43.6 and 43.7 kg/d, respectively). Milk protein yield was also increased in cows fed SBM+F. No differences were observed with body weight or condition score gain throughout the trial. Apparent total-tract digestibility of fiber was decreased in cows fed SBM+F, likely as a result of increased intake. Responses are consistent with previous research in our laboratory that demonstrated a decrease in ruminal CP degradation, leading to an increase in metabolizable protein supply in the small intestine. The fermentation by-product might be useful in diets containing adequate amounts of RDP from soybean meal or alfalfa. The results from this experiment demonstrate beneficial milk performance responses to fermentation by-product when fed with a source of RDP.

Effect of enzymatically hydrolyzed yeast on health and performance of transition dairy cattle.

The objective of this study was to evaluate the effect of supplementing enzymatically hydrolyzed yeast (EHY; Celmanax Arm and Hammer Animal Nutrition, Princeton, NJ) on transition dairy cattle. Forty multiparous Holstein cows were blocked by predicted transmitting ability and randomly assigned to 1 of 2 treatments (EHY, n = 20; or control, CON, n = 20) from 21 d before expected calving to 60 d postpartum. The EHY cows received 56 and 28 g/d in close-up and lactating diets, respectively. Dry matter intake, health events, milk production parameters, feed efficiency, colostrum quality, reproductive parameters, body weight, and body condition score were monitored. Fecal samples collected on -21, -14, -7, 0, 1, 3, 5, 7, and 14 d relative to calving were analyzed for total coliforms, Clostridium perfringens, Salmonella, and Escherichia coli O157:H7. Blood samples were collected at 7, 14, and 21 d postpartum for analysis of ß-hydroxybutyrate. Sterile quarter milk samples collected at dry-off, calving, and wk 1, 2, and 3 of lactation were analyzed for milk pathogens and somatic cell count. Pre- or postpartum dry matter intake, body weight, body condition score, milk yield, and milk protein and fat yields did not differ among treatments. Milk fat and protein concentrations were greater in EHY cows than CON cows. ß-Hydroxybutyrate and health events were not different among treatments. The presence of fecal C. perfringens did not differ prepartum, but was lower in EHY cows postpartum. Milk pathogens and total intramammary infections did not differ between treatments at dry-off, calving, wk 1, or wk 2, but more EHY cows were infected with Staphylococcus sp. during wk 3 than CON cows. The EHY cows showed heat earlier than CON cows, but no other reproductive parameters were affected. The EHY supplementation during the transition period did not affect dry matter intake, milk yield, health events, or reproductive parameters but did increase milk protein and fat concentrations.

Effects of a commercial fermentation byproduct or urea on milk production, rumen metabolism, and omasal flow of nutrients in lactating dairy cattle.

The objective of this study was to evaluate the effects of a fermentation byproduct on rumen fermentation and microbial yield in high producing lactating dairy cattle. Eight ruminally cannulated multiparous Holstein cows averaging (mean ± standard deviation) 60 ± 10 d in milk and 637 ± 38 kg of body weight were assigned to 1 of 2 treatment sequences in a switchback design. Treatment diets contained (dry matter basis) 44% corn silage, 13% alfalfa silage, 12% ground corn, and 31% premix containing either a control mix of urea and wheat middlings (CON) or a commercial fermentation byproduct meal (Fermenten, Arm and Hammer Animal Nutrition, Princeton, NJ) at 3% diet inclusion rate (EXP). Diets were formulated to be isonitrogenous and isocaloric, with similar levels of neutral detergent fiber and starch. The trial consisted of three 28-d experimental periods, where each period consisted of 21 d of diet adaptation and 7 d of data and sample collection. Omasal nutrient flows were determined using a triple-marker technique and double-labeled 15N15N-urea. The EXP diet provided 18 g/d more nonammonia N versus the CON diet, representing 3.0% of total N intake. Energy-corrected milk yield (41.7 and 43.1 kg/d for CON and EXP, respectively), milk fat, and protein yield and content did not differ between treatments. Total dry matter intake was similar between treatments (25.5 and 26.4 kg/d for CON and EXP, respectively). Ammonia N concentration and pool size in the rumen was greater in cows fed the EXP diet. No differences were observed in rumen or total-tract dry matter, organic matter, or neutral detergent fiber digestibility. Ruminal degradation of feed N was 15% lower in cows fed EXP diets, resulting in differences in omasal N flows. Results demonstrated the fermentation byproduct meal had a sparing effect on degradable feed protein, but did not increase microbial N flow from the rumen.

Rumen digestion kinetics, microbial yield, and omasal flows of nonmicrobial, bacterial, and protozoal amino acids in lactating dairy cattle fed fermentation by-products or urea as a soluble nitrogen source.

The objective of this study was to evaluate the effect of a fermentation by-product on rumen function, microbial yield, and composition and flows of nutrients from the rumen in high-producing lactating dairy cattle. Eight ruminally cannulated multiparous Holstein cows averaging (mean ± standard deviation) 60 ± 10 d in milk and 637 ± 38 kg of body weight were randomly assigned to 1 of 2 treatment sequences in a switchback design. Treatment diets contained (dry matter basis) 44% corn silage, 13% alfalfa silage, 12% ground corn, and 31% protein premix, containing either a control mix of urea and wheat middlings (CON) or a commercial fermentation by-product meal (Fermenten, Arm and Hammer Animal Nutrition, Princeton, NJ) at 3% diet inclusion rate (EXP). The trial consisted of three 28-d experimental periods, where each period consisted of 21 d of diet adaptation and 7 d of data and sample collection. A triple-marker technique and double-labeled 15N15N-urea were used to were used to measure protozoal, bacterial, and nonmicrobial omasal flow of AA. Rumen pool sizes and omasal flows were used to determine digestion parameters, including fractional rates of carbohydrate digestion, microbial growth, and yield of microbial biomass per gram of degraded substrate. Fermentation by-product inclusion in EXP diets increased microbial N and amino acid N content in microbes relative to microbes from CON cows fed the urea control. Microbial AA profile did not differ between diets. Daily omasal flows of AA were increased in EXP cows as a result of decreased degradation of feed protein. The inclusion of the fermentation by-product increased nonmicrobial AA flow in cows fed EXP versus CON. Average protozoal contribution to microbial N flow was 16.8%, yet protozoa accounted for 21% of the microbial AA flow, with a range of 8 to 46% for individual AA. Cows in this study maintained an average rumen pool size of 320 g of microbial N, and bacterial and protozoal pools were estimated at 4 different theoretical levels of selective protozoa retention. Fractional growth rate of all microbes was estimated to be 0.069 h-1, with a yield of 0.44 g of microbial biomass per gram of carbohydrate degraded. Results indicated that fermentation by-product can increase omasal flow of AA while maintaining adequate rumen N available for microbial growth and protein synthesis. Simulations from a developmental version of the Cornell Net Carbohydrate and Protein System indicated strong agreement between predicted and observed values, with some areas key for improvement in AA flow and bacterial versus protozoal N partitioning.

Meta-analysis of the effects of prepartum dietary cation-anion difference on performance and health of dairy cows.

The objectives were to use meta-analytic methods to determine the effects of changes in dietary cation-anion difference (DCAD) prepartum on productive performance and health of dairy cows. The literature was systematically reviewed, searching randomized experiments with transition cows that manipulated the prepartum DCAD or experiments with acidogenic diets in which dietary Ca, P, or Mg was manipulated. Forty-two experiments, including 134 treatment means and 1,803 cows, were included in the meta-analysis. Of those, 5 experiments with 15 treatment means reported responses for 151 nulliparous cows. Data collected included the mineral composition of prepartum diets, parity group prepartum, breed, days on treatment, and means and respective measure of variance for urine pH, dry matter intake (DMI), body weight, body condition, productive performance, concentrations of minerals and metabolites in blood, and incidence of diseases. Mixed effects meta-analyses were conducted weighting by the inverse of standard error of the means squared to account for the precision of each experiment. Models include the effects of DCAD, parity group prepartum, interaction between DCAD and parity group, and other covariates that showed significance in univariable analysis. Final models were selected based on parsimony and model fit. Reducing the prepartum DCAD reduced intake prepartum but improved intake postpartum in both parity groups. Interactions between DCAD and parity group occurred for yields of milk, fat-corrected milk (FCM), fat, and protein because reducing the DCAD improved those responses in parous cows; however, reducing the DCAD either had no effect on yields of milk and protein or reduced the yield of FCM and fat in nulliparous cows. The resulting equations from the statistical models predicted that reducing the DCAD from +200 to -100 mEq/kg would increase blood total Ca on the day of calving from 1.86 to 2.04 ± 0.05 mM, DMI postpartum 1.0 kg/d, and milk yield 1.7 kg/d in parous cows. The increased concentrations of blood total Ca at calving and postpartum explained the marked reduction in risk of milk fever in parous cows with a reduction in DCAD. As the DCAD decreased, the risk of retained placenta and metritis also decreased, resulting in fewer disease events per cow in both nulliparous and parous cows. Dietary concentrations of Ca, P, or Mg prepartum had no effect on DMI or yields of milk and FCM; however, increasing dietary Ca within the study range of 0.16 to 1.98% of dry matter tended to increase the risk of milk fever in parous cows regardless of DCAD fed. Collectively, results support the recommendation of prepartum acidogenic diets to result in a negative DCAD to parous cows with improvements in lactation performance and reduced risk of diseases; however, the range of DCAD fed did not allow for detection of an optimum value for postpartum performance. On the other hand, despite improvements in blood concentrations of Ca and reduction in uterine diseases with a reduction in DCAD fed to nulliparous cows, productive performance was either depressed or unaffected and the limited number of experiments did not provide sufficient evidence for a recommended DCAD for this group of cows.

Effects of prepartum dietary cation-anion difference intake on production and health of dairy cows: A meta-analysis.

Prepartum diets influence cow performance for weeks to months postpartum. This observation leads to questions about milk yield and physiological and health responses to diets with negative dietary cation-anion difference (DCAD). Further, responses to increased intake of a diet with lower DCAD (Eq/d) have not been explored using meta-analysis. Our objectives were to explore the effects of prepartum DCAD intake on metabolism and production and health as well as the potential for differences in intake of other macrominerals to influence responses to differences in DCAD intake using classical meta-analytical methods. Not all treated groups were fed a diet with negative DCAD, and the effect studied is that of reducing the DCAD. We hypothesized that reducing DCAD intake would improve Ca metabolism and postpartum performance. We used a maximum of 58 comparisons from 31 experiments and a total of 1,571 cows. Intakes of DCAD were 2.28 Eq/d and -0.64 Eq/d for the control, higher DCAD and treated, lower DCAD groups, respectively. Diets with lower DCAD reduced urine pH [standardized mean difference (SMD) = 1.90 and weighted mean difference (WMD) -1.23 pH]. Intake of lower DCAD decreased prepartum DMI (SMD = 0.23; WMD = 0.29 kg/d), increased postpartum DMI (SMD = 0.40; WMD = 0.63 kg/d), and increased milk yield (SMD = 0.172). However, we found an interaction with parity; diets with lower DCAD increased milk yield in parous cows (SMD = 0.29; WMD = 1.1 kg/d) but resulted in numerically lower milk yield in nulliparous cows (SMD = -0.20; WMD = 1.28 kg/d) compared with controls. The FCM yield increased with treatment (SMD = 0.12; WMD = 0.56 kg/d); however, yield of treated cows tended to be greater in parous cows but smaller for nulliparous cows compared with controls. Milk fat percentage, milk fat yield, and milk protein percentages were not affected by treatment, although milk protein yield tended to increase in cows fed the lower DCAD diet (SMD = 0.21; WMD = 0.02 kg/d). Treatment increased blood Ca (SMD = 0.53; WMD = 0.13 mM) and P (SMD = 0.40; WMD = 0.13 mM) on the day of calving and Ca postpartum (SMD = 0.36; WMD = 0.06 mM). Treated cows had smaller concentration of blood BHB before calving than controls (SMD = -0.39; WMD = -0.04 mM). Reducing DCAD in cows resulted in decreased risks of clinical hypocalcemia (risk ratio = 0.60) and retained placenta (risk ratio = 0.59), and reduced the odds of metritis (odds ratio = 0.46) and overall disease (OR = 0.61). We observed no effect on risk of abomasal displacement or mastitis and no effect of differences between treated and control cows in Ca intake (g/d) on the outcomes evaluated. A positive role for increased Mg intake between groups for increased milk fat yield and in reducing the risk of retained placenta was identified. Diets with lower DCAD improved performance of parous dairy cows, and our findings suggest a need for more studies on the effects of a lower DCAD on nulliparous transition cows.

Effect of dietary cation-anion difference on acid-base status and dry matter intake in dry pregnant cows.

The objective was to determine if the reduction in dry matter (DM) intake of acidogenic diets is mediated by inclusion of acidogenic products, content of salts containing Cl, or changes in acid-base status. The hypothesis was that a decrease in intake is mediated by metabolic acidosis. Ten primigravid Holstein cows at 148 ± 8 d of gestation were used in a duplicated 5 × 5 Latin square design. The dietary cation-anion difference (DCAD) of diets and acid-base status of cows were manipulated by incorporating an acidogenic product or by adding salts containing Cl, Na, and K to the diets. Treatments were a base diet (T1; 1.42% K, 0.04% Na, 0.26% Cl; DCAD = 196 mEq/kg); the base diet with added 1% NaCl and 1% KCl (T2; 1.83% K, 0.42% Na, 1.23% Cl; DCAD = 194 mEq/kg); the base diet with added 7.5% acidogenic product, 1.5% NaHCO3, and 1% K2CO3 (T3; 1.71% K, 0.54% Na, 0.89% Cl; DCAD = 192 mEq/kg); the base diet with added 7.5% acidogenic product (T4; 1.29% K, 0.13% Na, 0.91% Cl; DCAD = -114 mEq/kg); and the base diet with 7.5% acidogenic product, 1% NaCl, and 1% KCl (T5; 1.78% K, 0.53% Na, 2.03% Cl; DCAD = -113 mEq/kg). Periods lasted 14 d with the last 7 d used for data collection. Feeding behavior was evaluated for 12 h in the last 2 d of each period. Reducing the DCAD by feeding an acidogenic product reduced blood pH (T1 = 7.450 vs. T2 = 7.436 vs. T3 = 7.435 vs. T4 = 7.420 vs. T5 = 7.416) and induced a compensated metabolic acidosis with a reduction in bicarbonate, base excess, and partial pressure of CO2 in blood, and reduced pH and strong ion difference in urine. Reducing the DCAD reduced DM intake 0.6 kg/d (T1 = 10.3 vs. T4 = 9.7 kg/d), which was caused by the change in acid-base status (T2 + T3 = 10.2 vs. T4 + T5 = 9.6 kg/d) because counteracting the acidifying action of the acidogenic product by adding salts with strong cations to the diet prevented the decline in intake. The decline in intake caused by metabolic acidosis also was observed when adjusted for body weight (T2 + T3 = 1.75 vs. T4 + T5 = 1.66% BW). Altering the acid-base status with acidogenic diets reduced eating (T2 + T3 = 6.7 vs. T4 + T5 = 5.9 bouts/12 h) and chewing (T2 + T3 = 14.6 vs. T4 + T5 = 13.5 bouts/12 h) bouts, and extended meal duration (T2 + T3 = 19.8 vs. T4 + T5 = 22.0 min/meal) and intermeal interval (T2 + T3 = 92.0 vs. T4 + T5 = 107.7 min). Results indicate that reducing the DCAD induced a compensated metabolic acidosis and reduced DM intake, but correcting the metabolic acidosis prevented the decline in DM intake in dry cows. The decrease in DM intake in diets with negative DCAD was mediated by metabolic acidosis and not by addition of acidogenic product or salts containing Cl.

Effects of level of dietary cation-anion difference and duration of prepartum feeding on performance and metabolism of dairy cows.

The objectives were to evaluate the effects of feeding diets with 2 levels of negative dietary cation-anion differences (DCAD) during the last 42 or 21 d of gestation on performance and metabolism in dairy cows. The hypothesis was that extending feeding from 21 to 42 d and reducing the DCAD from -70 to -180 mEq/kg of dry matter (DM) would not be detrimental to performance. Holstein cows at 230 d of gestation were blocked by parity prepartum (48 entering their second lactation and 66 entering their third or greater lactation) and 305-d milk yield, and randomly assigned to 1 of 4 treatments arranged as a 2 × 2 factorial. The 2 levels of DCAD, -70 or -180 mEq/kg of DM, and 2 feeding durations, the last 21 d (short) or the last 42 d (long) prepartum resulted in 4 treatments, short -70 (n = 29), short -180 (n = 29), long -70 (n = 28) and long -180 (n = 28). Cows in the short treatments were fed a diet with DCAD of +110 mEq/kg of DM from -42 to -22 d relative to calving. After calving, cows were fed the same diet and production and disease incidence were evaluated for 42 d in milk, whereas reproduction and survival was evaluated for 305 d in milk. Blood was sampled pre- and postpartum for quantification of metabolites and minerals. Reducing the DCAD linearly decreased prepartum DM intake between -42 and -22 d relative to calving (+110 mEq/kg of DM = 11.5 vs. -70 mEq/kg of DM = 10.7 vs. -180 mEq/kg of DM = 10.2 ± 0.4), and a more acidogenic diet in the last 21 d of the dry period reduced intake by 1.1 kg/d (-70 mEq/kg of DM = 10.8 vs. -180 mEq/kg of DM = 9.7 ± 0.5 kg/d). Cows fed the -180 mEq/kg of DM diet had increased concentrations of ionized Ca in blood on the day of calving (-70 mEq/kg of DM = 1.063 vs. -180 mEq/kg of DM = 1.128 ± 0.020 mM). Extending the duration of feeding the diets with negative DCAD from 21 to 42 d reduced gestation length by 2 d (short = 277.2 vs. long = 275.3 d), milk yield by 2.5 kg/d (short = 40.4 vs. long = 37.9 ± 1.0 kg/d) and tended to increase days open because of reduced pregnancy per artificial insemination (short = 35.0 vs. long = 22.6%). Results suggest that increasing the duration of feeding diets with negative DCAD from 21 to 42 d prepartum might influence milk yield and reproduction of cows in the subsequent lactation, although yields of 3.5% fat- and energy-corrected milk did not differ with treatments. Reducing the DCAD from -70 to -180 mEq/kg of DM induced a more severe metabolic acidosis, increased ionized Ca concentrations prepartum and on the day of calving, and decreased colostrum yield in the first milking, but had no effects on performance in the subsequent lactation. Collectively, these data suggest that extending the feeding of an acidogenic diet beyond 21 d is unnecessary and might be detrimental to dairy cows, and a reduction in the DCAD from -70 to -180 mEq/kg of DM is not needed.

Associations between bone and energy metabolism in cows fed diets differing in level of dietary cation-anion difference and supplemented with cholecalciferol or calcidiol.

Bone-derived hormones play an important role in metabolism. This study examined the hypothesis that interactions between bone and energy metabolism, particularly those involving osteocalcin, are present in dairy cattle and have feedback mechanisms over time. Associations between metabolites in blood were examined in 32 Holstein cows blocked by parity and milk yield and randomly allocated to diets containing either 0.27 mg/kg dry matter (DM) calcidiol or cholecalciferol for an anticipated intake of 3 mg/d (120,000 IU/d) at 11 kg of DM, and positive (+130 mEq/kg DM) or negative (-130 mEq/kg DM) dietary cation-anion difference (DCAD) from 252 d of gestation to calving. Blood was sampled every 3 d, from 9 d prepartum to 30 d postpartum, and plasma concentrations of vitamin D3, 25-hydroxyvitamin D3, adiponectin, C-telopeptide of type 1 collagen (CTX1), glucose, insulin-like growth factor 1 (IGF1), insulin, undercarboxylated osteocalcin (uOC), and carboxylated osteocalcin (cOC) were determined. Feeding calcidiol compared with cholecalciferol increased plasma concentrations of 25-hydroxyvitamin D3 pre- (264.2 ± 8.0 vs. 61.3 ± 8.0 ng/mL) and postpartum (170.8 ± 6.2 vs. 51.3 ± 6.2 ng/mL) but decreased concentrations of vitamin D3 pre- (1.2 ± 0.6 vs. 14.5 ± 0.6 ng/mL) and postpartum (1.9 ± 0.4 vs. 3.2 ± 0.6 ng/mL). Prepartum, cows fed the negative DCAD diet had reduced concentrations of vitamin D3 and glucose compared with cows fed a positive DCAD. The combination of negative DCAD and cholecalciferol reduced IGF1 concentrations prepartum. The DCAD treatment had no effect on postpartum concentrations of metabolites. Nulliparous cows had increased concentrations of OC, CTX1, IGF1, glucose, and insulin compared with parous cows. Time series analysis identified associations between metabolites on the same day and over 3-d lags up to ±9 d that suggest feedback between 25-hydroxyvitamin D3 and vitamin D3 in the negative lags, indicating that 25-hydroxyvitamin D3 may exert feedback on vitamin D3 but not vice versa. We found evidence of a feedback mechanism between vitamin D3 and IGF1, with positive effect size (ES) on the same day and 3 d later, and negative ES 9 d later, that was more evident in cholecalciferol-fed cows. This suggests an important role of IGF1 in integrating bone metabolism with energy and protein metabolic pathways. Evidence of feedback was found between uOC and particularly cOC with IGF1, with positive ES on the same day but negative ES 6 d before and 6 d after. An association between uOC or cOC and IGF1 has not been previously identified in cattle and suggests that both uOC and cOC may have marked biological activity. Associations between OC and insulin identified in mice were not observed herein, although associations between OC and glucose were similar to those between IGF1 and glucose, supporting associations between glucose, OC, and IGF1. We provide further statistical evidence of crosstalk between vitamin D compounds, bone hormones, and energy metabolism in cattle. In particular, associations between uOC or cOC and IGF1 may provide links between prepartum diets and observations of prolonged increases in milk production and allow better control of peripartum metabolism.

Interaction of 5-hydroxy-l-tryptophan and negative dietary cation-anion difference on calcium homeostasis in multiparous peripartum dairy cows.

Hypocalcemia affects almost 50% of all dairy cows. Our laboratory has previously demonstrated that infusions of the serotonin precursor 5-hydroxy-l-tryptophan (5-HTP) increase circulating calcium concentrations in the Holstein transition cow. It is unknown whether feeding a negative dietary cation-anion difference (DCAD) diet alters the relationship between 5-HTP and hypocalcemia. The main objective of this study was to determine whether feeding a negative DCAD (-DCAD) diet before calving in conjunction with 5-HTP treatment could further diminish the magnitude of hypocalcemia at the time of calving. We used a randomized complete block design with a 2 × 2 factorial arrangement. Thirty-one multiparous Holstein cows were fed either a positive (+13 mEq/100 g) or negative (-13 mEq/100 g) DCAD diet 21 d before parturition and were intravenously infused daily with saline or 5-HTP (1 mg/kg) starting 7 d before the estimated date of parturition. Cows were blocked by parity and were randomly assigned to 1 of 4 treatment groups: positive DCAD plus saline, positive DCAD plus 5-HTP, negative DCAD plus saline, and negative DCAD plus 5-HTP, resulting in n = 8 per group. Total calcium (tCa), ionized calcium (iCa), and feed intake were recorded. The iCa was elevated prepartum in the -DCAD/5-HTP group compared with the other treatment groups as well as on d 0 and 1 postpartum. Although differences in tCa were not significant across the pre- or postpartum periods, tCa was numerically higher on d 0 and significantly higher on d 1 in -DCAD/5-HTP cows compared with all other groups. Prepartum the -DCAD/5-HTP treatment group ate less than the other treatment groups; however, postpartum dry matter intake differences were not significant. These findings demonstrate that feeding a -DCAD diet in conjunction with 5-HTP prepartum can increase postpartum circulating iCa concentrations and therefore diminish the magnitude of hypocalcemia at the time of parturition.

Effects of prepartum dietary cation-anion difference and source of vitamin D in dairy cows: Lactation performance and energy metabolism.

The objectives of this experiment were to evaluate the effects of feeding diets with 2 dietary cation-anion difference (DCAD) levels and supplemented with either cholecalciferol (CH) or calcidiol (CA) during late gestation on lactation performance and energetic metabolism in dairy cows. The hypothesis was that combining a prepartum acidogenic diet with calcidiol supplementation would benefit peripartum Ca metabolism and, thus, improve energy metabolism and lactation performance compared with cows fed an alkalogenic diet or cholecalciferol. Holstein cows at 252 d of gestation were blocked by parity (28 nulliparous and 51 parous cows) and milk yield within parous cows, and randomly assigned to 1 of 4 treatments arranged as a 2 × 2 factorial, with 2 levels of DCAD (positive, +130, and negative, -130 mEq/kg) and 2 sources of vitamin D, CH or CA, fed at 3 mg per 11 kg of diet dry matter (DM). The resulting treatment combinations were positive DCAD with CH (PCH), positive DCAD with CA (PCA), negative DCAD with CH (NCH), or negative DCAD with CA (NCA), which were fed for the last 21 d of gestation. After calving, cows were fed the same lactation diet. Body weight and body condition were evaluated prepartum and for the first 49 d postpartum. Blood was sampled thrice weekly prepartum, and on d 0, 1, 2, 3, and every 3 d thereafter until 30 d postpartum for quantification of hormones and metabolites. Lactation performance was evaluated for the first 49 d postpartum. Feeding a diet with negative DCAD reduced DM intake in parous cows by 2.1 kg/d, but no effect was observed in nulliparous cows. The negative DCAD reduced concentrations of glucose (positive = 4.05 vs. negative = 3.95 mM), insulin (positive = 0.57 vs. negative = 0.45 ng/mL), and insulin-like growth factor-1 (positive = 110 vs. negative = 95 ng/mL) prepartum. Treatments did not affect DM intake postpartum, but CA-supplemented cows tended to produce more colostrum (PCH = 5.86, PCA = 7.68 NCH = 6.21, NCA = 7.96 ± 1.06 kg) and produced more fat-corrected milk (PCH = 37.0, PCA = 40.1 NCH = 37.5, NCA = 41.9 ± 1.8 kg) and milk components compared with CH-supplemented cows. Feeding the negative DCAD numerically increased yield of fat-corrected milk by 1.0 kg/d in both nulliparous and 1.4 kg/d in parous cows. Minor differences were observed in postpartum concentrations of hormones and metabolites linked to energy metabolism among treatments. Results from this experiment indicate that replacing CH with CA supplemented at 3 mg/d during the prepartum period improved postpartum lactation performance in dairy cows.