Effects of feeding sulfate trace minerals above recommendations on nutrient digestibility, rumen fermentation, lactational performance, and trace mineral excretion in dairy cows.

Most trace minerals (TM) are fed above dairy cow requirements in commercial herds but their fate and effects on dairy cows have not been well documented. In this study, we evaluated the effects of feeding short-term sulfate TM above recommendations on apparent total-tract digestibility of nutrients, rumen fermentation characteristics, serum concentrations, milk yield and composition as well as milk, fecal, and urinary TM excretion in mid-lactation dairy cows. Eight multiparous Holstein cows [average body weight: 684 (SD: 29) kg at 82 (SD: 10) days in milk] in a quadruple 2 × 2 crossover design were fed a basal diet, differing in sulfate TM supplement concentrations, to provide either 0.11, 17, and 63 (control; CON) or 0.95, 114, and 123 (high trace minerals; HTM) mg of dietary Co, Mn, and Zn/kg of dry matter, respectively. Each experimental period had a 21-d adaptation to the diet, followed by a 10-d sample collection period. Feed ingredients and total feces and urine were collected during 4 consecutive d and rumen fluid was collected 0, 1, 2, 4, and 6 h relative to feeding. Milk yield was recorded daily and milk samples were collected on 4 consecutive milkings. Ingestion of Co, Mn, and Zn was higher for HTM compared with CON group by 216, 233, and 93%, respectively. Dry matter intake averaged 25.0 (SE = 0.6) kg/d, and apparent total-tract digestibility of major nutrients was similar between treatments. There was no measurable effect of HTM on ruminal pH, major volatile fatty acids, and protozoa counts. Isovalerate molar proportion was 9.4% greater for HTM compared with CON group. Neither milk yield (43.5 kg/d; SE = 0.8) nor milk fat and protein concentrations differed between treatments. Milk urea nitrogen concentration was significantly higher for HTM (11.7 mg/dL) compared with CON group (9.7 mg/dL; SE = 0.7). Fecal excretion of Co, Mn, and Zn increased by 223, 198, and 75%, respectively, for HTM compared with CON group. Urinary excretions of TM were marginal compared with feces, and only urinary Co and Mn were significantly higher for HTM than CON cows as similarly obtained for serum Co and Mn concentrations. Milk TM yields were not modified by treatments. In summary, short-term dietary sulfate TM supply over the recommendation did not improve cow performance but significantly increased fecal TM excretion, which could have impacts on TM accumulation in soils where manure is applied and could potentially result in leaching into nearby watersheds. Further studies are needed to evaluate the impact of high fecal TM excretion on the environment using the One Health approach. Moreover, the impacts of TM oversupply on milk production and cow health should be evaluated by long-term experiments.

Longitudinal Data to Assess Relationships among Plasma Folate, Vitamin B12, Non-esterified Fatty Acid, and ß-Hydroxybutyrate Concentrations of Holstein Cows during the Transition Period.

It is well established that the plasma metabolite profile changes during metabolic dysfunction, such as elevated non-esterified fatty acid (NEFA) release when body reserve mobilization is excessive in early lactation cows. Relationships between changes in plasma concentrations of metabolites caused by a metabolic impairment and the status of vitamins, such as folates and vitamin B12, have barely been studied in cattle. This study was undertaken to assess relationships between peripartum plasma concentrations of folates, vitamin B12, NEFA, and beta-hydroxybutyrate (BHB). Longitudinal data of 48 multiparous Holstein cows from 5 studies were taken from days -14 to 21 relative to calving. Blood samples were taken weekly before calving and either twice or thrice per week postpartum, and plasma was analyzed for folate, vitamin B12, NEFA, and BHB concentrations. Postpartum plasma NEFA and BHB concentrations were negatively related to plasma folate concentration at days -14 and -7 relative to parturition, whereas the opposite relationship was noted for the plasma vitamin B12:folate ratio. The plasma folate and NEFA areas under the curve from the whole studied period were negatively associated, and the opposite was observed with the association between the plasma vitamin B12:folate ratio and NEFA as well as the BHB areas under the curve. The results suggest that there is an increased use of folate for metabolic functions during elevated concentrations of plasma NEFA and BHB. Future research should focus on finding an optimal plasma vitamin B12:folate ratio to favor cow health during the challenging period of parturition.

A survey of Canadian dairy nutritionists to assess current trace element formulation practices.

The purpose of this study was to identify current practices and perceptions around trace element feeding for dairy cows through a Canadian dairy nutritionist survey. An online survey with 23 questions was used to collect data from Canadian dairy nutritionists with the help of professional associations and social media. The survey was active from November 2021 to April 2022. The first 7 questions collected descriptive information on respondents, and the subsequent 16 questions focused on trace element feeding. A total of 92 participants from all over Canada filled out the survey, and about 26% of Canadian herds and cows were represented by these respondents. The participants had diverse views on the importance of diet formulations for trace elements to optimize cow health and productivity, with perceptions varying from very important to not important. In comparison, macronutrients and selenium were consistently rated as very important by between 58% and 74% of respondents. Software reference values were used by 54%, 72%, and 73% of participants to estimate trace element concentrations of forages, cereals, and protein sources, respectively, highlighting the importance of regularly updating the feed library of the software. More than 60% of nutritionists participating in this study had intentionally formulated diets above trace element software recommendations, considered mineral interactions occurring in the rumen, and used a trace element source known for its better bioavailability (e.g., organic, chelate) when they formulated diets. Herds with more than 80 cows were more likely to be given trace element supplements known for their greater bioavailability. The most used supplement with enhanced bioavailability was selenium. In addition, different trace element feeding strategies pertaining to different stages of lactation and breeds were reported. This finding can be explained by the absence of clear recommendations on trace element feeding by breed. The participants who adjusted trace element feeding according to the stages of lactation considered the transition period as the most challenging period, and they identified the need for a source of trace element known for its greater bioavailability for this period. Further research should aim to identify environmental risk of trace element overfeeding using the One Health approach. Moreover, strategies to avoid trace element overfeeding should be evaluated.

Effects of replacing inorganic salts of trace minerals with organic trace minerals in the pre- and postpartum diets on mineral status, antioxidant biomarkers, and health of dairy cows.

Our objectives were to evaluate the effects of complete replacement of supplementary inorganic salts of trace minerals (ITM; cobalt (Co), copper (Cu), manganese (Mn), zinc (Zn) sulfates and sodium (Na) selenite) by organic trace minerals (OTM; Co, Cu, Mn, Zn proteinates, and selenized yeast) in both pre- and postpartum diets on trace minerals (TM) concentrations in body fluids and liver, antioxidant and inflammation biomarkers in blood, and postpartum health of dairy cows. Pregnant cows were blocked by parity and body condition score and randomly assigned to ITM (n = 136) or OTM (n = 137) 45 d before expected calving. Both groups received the same pre- and postpartum diets except for the source of supplementary TM. The day of calving was considered study d 0 and blood was collected on d -45, -21, -14, -10, -7, -3, 0, 3, 7, 10, 14, 23, 65, and 105 for analyses of TM and biomarkers. Concentrations of TM were also investigated in the liver (d 105), milk (d 7, 23, 65, 105), urine (d -21, 21, 65, 105), ruminal fluid and feces (d -21, 21, 65). Incidence of clinical and subclinical health conditions were evaluated. Complete replacement of ITM by OTM resulted in greater concentration of selenium (Se) in serum (0.084 vs. 0.086 µg/mL; P < 0.01), milk (0.24 vs. 0.31 µg/g; P < 0.01), and ruminal fluid (0.54 vs. 0.58 µg/g; P = 0.06), and reduced concentration of Se in urine (1.54 vs. 1.23 µg/g; P<0.01). For concentration of Co in serum, an interaction between treatment and time was detected (P < 0.01). Cows supplemented with OTM had greater concentrations of Co on d -7 and 0 (0.30 vs. 0.33 ng/mL; P < 0.01) but lower concentrations of Co on d 23, 65, and 105 (0.34 vs. 0.31 ng/mL; P < 0.05), in addition to reduced concentration of Co in feces (1.08 vs. 0.99 µg/g; P = 0.04) and, for multiparous only, in urine (0.019 vs. 0.014 µg/g; P < 0.01). Cows supplemented with OTM had lower postpartum concentrations of glutamate dehydrogenase (20.8 vs. 17.8 U/L; P < 0.05) and higher albumin on d -10 (36.0 vs. 36.7 g/L; P = 0.04) and 23 (36.9 vs. 37.6 g/L; P = 0.03) relative to calving. Primiparous cows fed OTM had lower concentration of ceruloplasmin in plasma (55 vs. 51 mg/L; P ≤ 0.05). Cows supplemented with OTM had less incidence of lameness (14% vs. 7%; P = 0.05), elevated nonesterified fatty acids (NEFA) (61% vs. 44%; P < 0.01), and multiple metabolic problems (35% vs. 20%; P < 0.01). Despite the lack of differences in Cu, Mn, and Zn concentrations and antioxidant capacity, complete replacement of ITM by OTM altered concentrations of Se and Co, supported liver and hoof health, and reduced the risk of postpartum elevated NEFA.

Trace minerals (TM) are important for oxidative balance and immunity of cows. Different forms of TM are available for dietary supplementation of dairy cows. We tested whether replacing inorganic salts of TM by organic sources of TM in both pre- and postpartum diets improve TM concentration in body fluids and liver, antioxidant capacity in blood, and postpartum health of dairy cows. Despite the lack of difference in antioxidant capacity and in concentrations of Cu, Mn, and Zn, the complete replacement of inorganic salts by organic sources altered concentrations of Se and Co in circulation, and reduced the concentration of biomarkers associated with inflammation and liver damage, and the risk of lameness and postpartum metabolic problems.

Toward Precision Feeding Regarding Minerals: What Is the Current Practice in Commercial Dairy Herds in Québec, Canada?

This analysis is performed to obtain information on the current situation regarding phosphorus (P), cobalt (Co), copper (Cu), iron (Fe), manganese (Mn), and zinc (Zn) concentrations in cow diets of commercial dairy herds in Québec, Canada, and to compare them with National Research Council recommendations. Data are collected on 100 Holstein dairy herds in Québec, Canada, and 4430 cows were involved. Rations are analyzed for selected minerals and cow requirements relative to the recommendations were calculated. Median percentages of mineral recommendations fulfilled by forage were 55%, 196%, 54%, 776%, 181%, and 44% for P, Co, Cu, Fe, Mn, and Zn, respectively. Daily dietary concentrations of P, Cu, Mn, and Zn decreased as lactation progressed, whereas Co and Fe were stable throughout lactation. Phosphorus was the mineral fed the closest to the requirements, cows below 21 days in milk were even underfed by 11%. All studied trace minerals were fed in excess for the majority of cows. Cobalt was fed on average 480% above requirements regardless of the stage of lactation. For Cu, Fe, Mn, and Zn, rations for cows below 21 days in milk were fed 23% (95% confidence interval: 15-32), 930% (849-1019), 281% (251-314), and 35% (22-47) above the recommendations, respectively, and were closer to the requirements than after 21 days in milk. These results show that most nutritionists are aware that precision feeding regarding P is important to minimize detrimental environmental impacts of dairy production. However, some efforts should be made to limit trace mineral overfeeding to ensure environmental resiliency.

Refining Knowledge of Factors Affecting Vitamin B12 Concentration in Bovine Milk.

Milk is an excellent source of vitamin B12 (B12) for humans. Therefore, being able to guarantee a high and consistent concentration of this vitamin would enhance consumer perception of milk as a health food. The aim of the paper was to gather additional knowledge on factors that could explain B12 variation in cow milk through two observational studies: (1) to explore the relationship between milk B12 and ruminal conditions, such as pH and volatile fatty acid concentrations; and (2) to examine the impact of bedding on B12 concentrations in bulk tank milk. For study 1, a total of 72 milk and ruminal liquid samples were obtained from 45 Holstein cows fitted with ruminal cannula between 10 and 392 days of lactation. For study 2, bulk tank milk samples were obtained from 83 commercial herds; 26 herds used recycled manure solid bedding and 57 used straw bedding. Milk samples were analyzed for B12 using radioassay. Using principal component regression analysis, we observed that ruminal pH and the acetate:propionate ratio for cows receiving the early lactation ration were positively correlated with milk B12. Bedding did not influence milk B12 in bulk tanks, which averaged 4276 pg/mL. In conclusion, as B12 is synthesized by ruminal bacteria, optimizing ruminal conditions had a positive effect on milk B12, while bedding management had no influence.

Correlations between the Composition of the Bovine Microbiota and Vitamin B12 Abundance.

Vitamin B12 is synthesized by prokaryotes in the rumens of dairy cows-and this has implications in human nutrition since humans rely on consumption of dairy products for vitamin B12 acquisition. However, the concentration of vitamin B12 in milk is highly variable, and there is interest in determining what causes vitamin B12 variability. We collected 92 temporally linked rumen, fecal, blood, and milk sample sets from Holstein cows at various stages of lactation fitted with rumen cannula and attempted to define which bacterial genera correlated well with vitamin B12 abundance. The level of vitamin B12 present in each sample was measured, and the bacterial population of each rumen, fecal, and milk sample (n = 263) was analyzed by 16S rRNA-targeted amplicon sequencing of the V4 region. The bacterial populations present in the rumen, small intestine, and milk were highly dissimilar. Combined diet and lactation status had significant effects on the composition of the microbiota in the rumen and in the feces. A high ruminal concentration of vitamin B12 was correlated with the increased abundance of Prevotella, while a low ruminal concentration of vitamin B12 was correlated with increased abundance of Bacteroidetes, Ruminiclostridium, and Butyrivibrio The ultimate concentration of vitamin B12 is controlled by the complex interaction of several factors, including the composition of the microbiota. Bacterial consumption of vitamin B12 in the rumen may be more important in determining overall levels than bacterial production.IMPORTANCE In this paper, we examined the microbiome of the bovine rumen, feces, and milk and attempted to understand how the bacterial communities at each site affected the production and movement of vitamin B12 around the animal's body. It was determined that the composition of the bovine rumen microbiome correlates well with vitamin B12 concentration, indicating that the rumen microbiota may be a good target for manipulation to improve production of this important vitamin.

Milk ß-hydroxybutyrate concentration measured by Fourier-transform infrared and flow-injection analyses from samples taken at different times relative to milking.

Analysis of milk BHB concentration by Fourier-transform infrared (FTIR) spectrometry more frequently than regular milk testing could help dairy producers in decision making, particularly if it would be possible to use small hand-stripped samples (hereinafter simply called samples) taken between dairy herd improvement (DHI) test-samples analysed using DHI algorithms. The aim of this Research Communication was to evaluate milk BHB concentration and the prevalence of elevated milk BHB concentration analysed by FTIR spectrometry compared with flow-injection analysis (SKALAR) from samples taken at different times relative to the milking. A total of 293 early-lactation cows in 44 commercial dairy herds were involved in the study. Herds were visited once during the morning milking when a routine DHI test-sample was obtained using in-line milk samplers. Additional milk samples were taken by hand stripping as follows: (1) Just before connecting the milking machine; (2) immediately after removing the milking machine; (3) 3 h after milking and (4) 6 h after milking. Milk samples were analysed for BHB concentration by FTIR and SKALAR, the latter being the reference method. Milk BHB concentration from samples taken before milking was different between FTIR and SKALAR whereas no difference was noted for other sampling times, although milk BHB concentration rose as time after milking increased. Except for DHI test-samples for which prevalence was not different between analysis methods, prevalence of elevated milk BHB concentration (≥0.15 mmol/l) was greater for FTIR analysis. However, no difference in prevalence was observed between SKALAR and FTIR when using a threshold of ≥0.20 mmol/l. In summary, hand-stripped milk samples taken any time after removing the milking machine until 6 h after the milking can be recommended for FTIR analysis of elevated milk BHB concentration prevalence provided a threshold of 0.20 mmol/l is used.

Supplements of vitamins B9 and B12 affect hepatic and mammary gland gene expression profiles in lactating dairy cows.

BACKGROUND: A combined supplement of vitamins B9 and B12 was reported to increase milk and milk component yields of dairy cows without effect on feed intake. The present study was undertaken to verify whether this supplementation positively modifies the pathways involved in milk and milk component synthesis. Thus, by studying the transcriptome activity in these tissues, the effect of supplements of both vitamins on the metabolism of both liver and mammary gland, was investigated. For this study, 24 multiparous Holstein dairy cows were assigned to 6 blocks of 4 animals each according to previous 305-day milk production. Within each block, cows were randomly assigned to weekly intramuscular injections of 5 mL of either saline 0.9 % NaCl, 320 mg of vitamin B9, 10 mg of vitamin B12 or a combination of both vitamins (B9 + B12). The experimental period began 3 weeks before the expected calving date and lasted 9 weeks of lactation. Liver and mammary biopsies were performed on lactating dairy cows 64 ± 3 days after calving. Samples from both tissues were analyzed by microarray and qPCR to identify genes differentially expressed in hepatic and mammary tissues. RESULTS: Microarray analysis identified 47 genes in hepatic tissue and 16 genes in the mammary gland whose expression was modified by the vitamin supplements. Gene ontology (GO) categorizes genes in non-overlapping domains of molecular biology. Panther is one of the online GO resources used for gene function classification. It classifies the 63 genes according to Molecular Function, Biological Process and Protein Class. Most of the biological processes modulated by the vitamin supplements were associated to developmental process, protein metabolic process, transport and response to inflammation. In the liver, most of the genes modulated by the vitamin treatments involved protein metabolic process while developmental process appeared to be more affected by the treatments in mammary gland. Out of 25 genes analysed by qPCR, 7 were validated. CONCLUSION: The results indicate that several metabolic processes were modulated by the supplementation of vitamins in early-lactating dairy cows. In addition, the results suggest that the vitamin supplements promoted liver regeneration and reduced catabolism of lipids in early lactation.

Optimization and validation of a reversed-phase high performance liquid chromatography method for the measurement of bovine liver methylmalonyl-coenzyme a mutase activity.

BACKGROUND: Methylmalonyl-CoA mutase (MCM) is an adenosylcobalamin-dependent enzyme that catalyses the interconversion of (2R)-methylmalonyl-CoA to succinyl-CoA. In humans, a deficit in activity of MCM, due to an impairment of intracellular formation of adenosylcobalamin and methylcobalamin results in a wide spectrum of clinical manifestations ranging from moderate to fatal. Consequently, MCM is the subject of abundant literature. However, there is a lack of consensus on the reliable method to monitor its activity. This metabolic pathway is highly solicited in ruminants because it is essential for the utilization of propionate formed during ruminal fermentation. In lactating dairy cows, propionate is the major substrate for glucose formation. In present study, a reversed-phase high performance liquid chromatography (RP-HPLC) was optimized and validated to evaluate MCM activity in bovine liver. The major aim of the study was to describe the conditions to optimize reproducibility of the method and to determine stability of the enzyme and its product during storage and processing of samples. RESULTS: Specificity of the method was good, as there was no interfering peak from liver extract at the retention times corresponding to methylmalonyl-CoA or succinyl-CoA. Repeatability of the method was improved as compared to previous RP-HPLC published data. Using 66 µg of protein, intra-assay coefficient of variation (CV) of specific activities, ranged from 0.90 to 8.05% and the CV inter-day was 7.40%. Storage and processing conditions (frozen homogenate of fresh tissue vs. fresh homogenate of tissue snapped in liquid nitrogen) did not alter the enzyme activity. The analyte was also stable in liver crude extract for three frozen/thawed cycles when stored at -20°C and thawed to room temperature. CONCLUSIONS: The improved method provides a way for studying the effects of stages of lactation, diet composition, and physiology in cattle on MCM activity over long periods of time, such as a complete lactation period. Interestingly, this sensitive and accurate method could benefit the study of the cobalamin status in experimental studies and clinical cases.