Effect of dietary phosphorus deprivation during the dry period on the liver transcriptome of high-yielding periparturient dairy cows.

Although dietary phosphorus (P) deprivation extending from the dry period into early lactation impairs health and productivity of cows, restricting dietary P supply during the dry period not only appears to be innocuous but rather effectively mitigates hypocalcemia during the first wk of lactation. To investigate possible negative metabolic effects of P deprivation during the dry period, the present study tested the hypothesis that restricted dietary P supply during the dry period alters the liver transcriptome of dairy cows during the periparturient period. Thirty late-pregnant multiparous Holstein-Friesian dairy cows entering their second, third, or fourth lactation were assigned to either a dry cow ration with low (LP, 0.16% P in DM) or adequate P content (AP, 0.35% in DM) during the last 4 wk of the dry period (n = 15/group). Liver transcriptomics, which was carried out in a subset of 5 second-parity cows of each group (n = 5), and determination of selected hormones and metabolites in blood of all cows, was performed ∼1 wk before calving and on d 3 postpartum. Liver tissue specimens and blood samples were obtained by a micro-invasive biopsy technique from the right tenth intercostal space and puncture of a jugular vein, respectively. One hundred seventy-five hepatic transcripts were expressed differentially between LP versus AP cows in late pregnancy, and 165 transcripts differed between LP versus AP cows in early lactation (fold change >1.3 and <-1.3, P < 0.05). In late pregnancy, the enriched biological processes of the upregulated and the downregulated transcripts were mainly related to immune processes and signal transduction (P < 0.05), respectively. In early lactation, the enriched biological processes of the upregulated and the downregulated transcripts were involved in mineral transport and biotransformation (P < 0.05), respectively. The plasma concentrations of the hormones and acute-phase proteins (progesterone, insulin-like growth factor 1, serum amyloid α, haptoglobin, and 17ß-estradiol) determined were not affected by P supply. These results suggest that P deprivation during the dry period moderately affects the liver transcriptome of cows in late pregnancy and early lactation, and causes no effects on important plasma hormones and acute-phase proteins indicating no obvious impairment of health or metabolism of the cows.

Evaluation of the relationship between ionized and total calcium concentrations in blood during the first week of lactation in dairy cows.

BACKGROUND: Diagnosis of subclinical hypocalcemia in cattle is based on concentration of total Ca ([TCa]) in serum or plasma below the reference range, with [TCa] serving as proxy for the concentration of ionized Ca ([iCa]). HYPOTHESIS/OBJECTIVES: To investigate the relation between [iCa] and [TCa] in periparturient cows over time, and its association with various metabolites. ANIMALS: Thirty periparturient dairy cows. METHODS: Prospective observational study. Blood from periparturient cows was obtained between 4 days before and 7 days after calving. Samples underwent blood gas analysis and blood biochemical analysis. The ratio of [iCa] : [TCa] was computed. Repeated measures linear regression analyses, spearman correlation-, Deming regression- and receiver operating characteristic (ROC) analyses were conducted. RESULTS: Correlations between [iCa] and [TCa] ranged from r = 0.55 to r = 0.84. The diagnostic value of [TCa] to identify cows with [iCa] below the arbitrary cut-off of 1.1 mmol/L was weak in particular during the first 24 hours of lactation, but continuously improved toward the end of the study with areas under the ROC curve increasing from 0.64 at d 0 to 0.93 at d +4 after calving. Strongest association with [iCa] : [TCa] were found for albumin (r2 = 0.58, P < .0001), pCO2 (r2 = 0.45, P = .0003), the standard [HCO3 ] (r2 = 0.22, P = .01), lactate (r2 = 0.16, P = .04) and [NEFA] (r2 = 0.15, P = .05). CONCLUSION AND CLINICAL IMPORTANCE: The [TCa] is of limited value to identify cows with subnormal [iCa] in the first hours and days of lactation, a finding apparently attributable to the increased variation of a number of metabolic variables that affect the ratio of [iCa] : [TCa].

Effects of oral or parenteral iron supplementation on haematological parameters, blood iron status and growth in newborn calves fed milk replacer.

BACKGROUND: This study aimed to assess the influence of early life iron supplementation on the haematological parameters of calves fed milk replacer on a high plane of nutrition. METHODS: Thirty calves were allocated to receive either a sham treatment (CON), injection of 1000 mg Fe3+ (INJ) or oral administration of 1050 mg Fe3+ (ORAL), all administered less than 1 hour after birth. Blood was obtained before treatment, on days 1, 3, 5 and 7 of life and once weekly until week 9. Samples were analysed for haemoglobin (Hb), packed cell volume (PCV) and estimated transferrin saturation (%TSAT). RESULTS: The Hb and PCV of calves in the INJ and ORAL groups were above the values of those in the CON group throughout the study. Hb and PCV of the CON group remained within the reference range for calves. The %TSAT exceeded reported ranges in the ORAL group in the first week. Values for the CON group were below the INJ and ORAL groups throughout the entire study. Average daily gain did not differ between treatments. LIMITATIONS: The sample size was insufficient to assess the influence of iron supplementation on disease development. Furthermore, the study was set in a controlled environment and not performed under field conditions. CONCLUSIONS: Oral and subcutaneous iron administration had similar effects on haematological development but resulted in numerically different transferrin saturation. Control animals showed lower Hb and PCV but did not develop overt anaemia.

Influence of birth weight, sex and disinfection on the involution of umbilical structures in calves.

BACKGROUND: Umbilical disinfection for calves is often recommended, but scientific studies do not always show a positive effect on disease prevention. In addition, there is little to no metric data on umbilical involution or on factors influencing the size of umbilical structures. Therefore, the aim of this study was to investigate the physiological involution of umbilical structures and the effect of two different umbilical treatment methods. METHODS: After birth, calves' umbilical cord stumps were treated with either an iodine tincture (n = 43) or an antibiotic spray (n = 41) or were left untreated (n = 42). On days 0, 1, 3, 7 and 14 after birth, calves were examined and external umbilical structures were measured. In addition, on day 14 (±2 days), a sonographic examination of umbilical structures was conducted. RESULTS: The treatment applied had no significant effect on the size of umbilical structures during the first 14 days of life. However, the diameter of the umbilical stalk was larger in male calves and calves with a higher birth weight than in female calves and calves with a lower birth weight. LIMITATION: The study was carried out on only one farm, so the generalisability of the findings may be limited. CONCLUSION: Umbilical structures showed significant involution over time and were influenced by birth weight and sex.

A Pilot Study on Across-Generation Impacts of Maternal Heat Stress on Blood Metabolites of Female Holstein Dairy Calves.

Heat stress (HS) during late gestation implies unfavorable effects on dairy cows and their in-utero heat stressed offspring. The objective of the present study was to elucidate the effect of intrauterine (maternal) HS during the last week of gestation on blood metabolite concentrations of female dairy calves during their first week of life. We defined the mean temperature humidity index (mTHI) during the last gestation week of ≥60 as threshold for maternal HS. In this regard, we compared differences in metabolite concentrations of maternally heat stressed (MHSCALVES) (n = 14) and not heat stressed (NMHSCALVES) (n = 33) calves. We identified 15 metabolites from five different biochemical classes (phosphatidylcholines, cholesteryl esters, sphingomyelins, cresols and hexoses) as potential biomarkers for maternal HS in calves. The plasma concentrations of all significantly affected metabolites were lower in MHSCALVES when compared to NMHSCALVES. The effect of maternal HS during the last week of gestation on blood metabolite concentrations of the female offspring during the first week after birth might be due to HS induced intergenerational physiological alterations, impaired colostrum quality or epigenetic modifications of the calf genome. The results of this pilot study should be validated in ongoing fully standardized studies.

Effect of Dietary Phosphate Deprivation on Red Blood Cell Parameters of Periparturient Dairy Cows.

Postparturient hemoglobinuria is a sporadic disease characterized by intravascular hemolysis and hemoglobinuria in early lactating dairy cows. The condition has empirically been associated with phosphorus (P) deficiency or hypophosphatemia; however, the exact etiology remains obscure. This paper summarizes two controlled studies investigating the effect of P deprivation during the transition period. In Study I, 36 late pregnant dairy cows were randomly assigned to either a diet with low, or adequate, P content from four weeks before calving to four weeks after calving. In Study II, 30 late pregnant dairy cows were again assigned to either a diet with low, or adequate, P for the last four weeks before calving only. Pronounced hypophosphatemia developed during periods of restricted P supply. In early lactation, a subtle decline of the red blood cell count occurred independently of the dietary P supply. In Study I, anemia developed in 11 cows on deficient P supply, which was associated with hemoglobinuria in five cases. Neither erythrocyte total P content nor osmotic resistance of erythrocytes were altered by dietary P deprivation. Restricted dietary P supply, particularly in early lactation, may lead to postparturient hemoglobinuria, but more frequently causes clinically inapparent hemolysis and anemia in cows.

Liver phosphorus content and liver function in states of phosphorus deficiency in transition dairy cows.

Phosphorus (P) deficiency in early lactating dairy cows is receiving increased attention because of incentives aiming at curtailing environmental pollution with P by reducing dietary P in ruminant diets. An in-vitro study using bovine hepatocytes incubated for 7 days with phosphate (Pi) concentrations of 0.9, 1.8 or 2.7 mmol/L, and an in-vivo study feeding late pregnant dairy cows diets with either adequate (0.28% and 0.44% in DM ante-partum and post-partum respectively) or low P content (0.15% and 0.20% in DM ante-partum and post-partum respectively) from 4 weeks before to 4 weeks after calving were conducted to explore effects of P deprivation on liver function. In vitro the relative abundance of mRNA of key enzymes of the carbohydrate metabolism in incubated hepatocytes and liver metabolites in culture medium were determined. In vivo health and productivity of experimental cows on low and adequate dietary P supply were monitored, and liver tissue and blood samples were obtained repeatedly. Liver tissue was assayed for its triacylglycerol-, mineral and water content as well as for the relative abundance of mRNA of enzymes of the carbohydrate-, fat- and protein metabolism. Reduced Pi-availability was not associated with altered enzyme transcription rates or metabolic activity in-vitro. The most prominent clinical finding associated with P deprivation in-vivo was feed intake depression developing after the first week of lactation. Accordingly cows on low P diets had lower milk yield and showed more pronounced increases in liver triacylglycerol after calving. Although the liver P content decreased in P deficient cows, neither negative effects on enzyme transcription rates nor on blood parameters indicative of impaired liver metabolic activity or liver injury were identified. These results indicate the P deprivation only indirectly affects the liver through exacerbation of the negative energy balance occurring as P deficient cows become anorectic.

Changes of the erythrocyte phenotype and blood biochemistry in dairy calves during the first ten weeks of age.

BACKGROUND: Bovine erythrocytes undergo important changes in their morphology and chemical composition during the first weeks of age, which must be understood to accurately interpret hematology results in calves. The objectives of this prospective cohort study were to describe physiological changes of calf erythrocytes and to investigate mechanisms potentially causing these changes. METHODS: Blood samples from 30 clinically healthy dairy calves were obtained from birth to the tenth week of age in weekly intervals. Hematological and plasma biochemical parameters as well as the mineral electrolyte content of erythrocytes were determined and followed over time. The changes of parameters characterizing the erythrocyte phenotype over time were compared to the changes of plasma and erythrocyte biochemical parameters and possible associations were investigated using correlation and stepwise regression analyses. RESULTS: Although the erythrocyte mean corpuscular volume (MCV) declined from 43.6 ± 3.7 fL to 35.6 ± 3.2 fL between the first and seventh week, the red blood cell count (RBC) increased from 7.2 ± 1.1 × 1012/L to 9.3 ± 1.0 × 1012/L until the fifth week of age. The blood hemoglobin (Hb) concentration increased from 0.96 ± 0.16 g/L to 1.16 ± 0.11 g/L in the first three weeks of age and remained at this level until the end of the study. Changes in MCV were accompanied by a decline of the erythrocyte potassium content (KERY) from 91.9 ± 13.5 to 24.6 ± 7.2 mmol/L and a concomitant increase of the erythrocyte sodium content from 45.0 ± 32.0 to 102.7 ± 26.5 mmol/L. MCV was found to be associated with KERY, the primary determinant of the intra-erythrocyte osmotic pressure from the sixth week of age and with blood hemoglobin, the primary determinant of the intra-erythrocyte oncotic pressure from the eighth week of age, when KERY, blood Hb and MCV already had reached or approached normal levels of adult cattle. The plasma iron concentration was not found to be associated to any of the studied hematological parameters. CONCLUSION: A volume reduction of 20% in bovine neonatal erythrocytes is a physiological change occurring during the first weeks of age and is neither associated with sideropenia nor with anemia in healthy calves. The mechanism driving the observed erythrocyte volume change could not be identified. Results of the correlation and regression analyses indicate that changes in intra-erythrocyte osmotic or oncotic pressure are improbable underlying causes. Results reported here show that KERY is an unreliable indicator for the K homeostasis of the intracellular space in neonatal calves and that a decrease in MCV in early life per-se is an unreliable indicator for the development of microcytic anemia.

Suitability of oral administration of monosodium phosphate, disodium phosphate, and magnesium phosphate for the rapid correction of hypophosphatemia in cattle.

BACKGROUND: Hypophosphatemia is commonly associated with disease and decreased productivity in dairy cows particularly in early lactation. Oral supplementation with phosphate salts is recognized as suitable for the rapid correction of hypophosphatemia. Little information is available about the differences in efficacy between salts used for oral phosphorus supplementation. OBJECTIVES: Comparison of efficacy of oral administration of NaH2 PO4 , Na2 HPO4 , and MgHPO4 in treating hypophosphatemia in cattle. ANIMALS: 12 healthy dairy cows in the fourth week of lactation in their second to fifth lactation. METHODS: Randomized clinical study. Phosphorus deficient, hypophosphatemic cows underwent a sham treatment and were afterwards assigned to 1 of 3 treatments-NaH2 PO4 , Na2 HPO4 , or MgHPO4 (each provided the equivalent of 60 g of phosphorus). Blood samples were obtained immediately before and repeatedly after treatment. RESULTS: Treatment with NaH2 PO4 and Na2 HPO4 resulted in rapid and sustained increases of plasma phosphate concentrations ([Pi]). Significant effects were apparent within 1 hour (NaH2 PO4 : P = .0044; Na2 HPO4 : P = .0077). Peak increments of plasma [Pi] of 5.33 mg/dL [5.26-5.36] and 4.30 mg/dL [3.59-4.68] (median and interquartile range) were reached after 7 and 6 hours in animals treated with NaPH2 PO4 and Na2 HPO4 , respectively, whereas treatment with MgHPO4 led to peak increments 14 hours after treatment (3.19 mg/dL [2.11-4.04]). CONCLUSIONS AND CLINICAL IMPORTANCE: NaH2 PO4 and Na2 HPO4 are suitable to rapidly correct hypophosphatemia in cattle. Because of the protracted and weaker effect, MgHPO4 cannot be recommended for this purpose. Despite important differences in solubility of NaH2 PO4 and Na2 HPO4 only small plasma [Pi] differences were observed after treatment.

Is the metabolism of 25-hydroxyvitamin D3 age-dependent in dairy cows?

It has recently been demonstrated that prepartum administered 25-hydroxyvitamin D3 (25-OHD3) is a promising candidate to assist the maintenance of peripartal calcium homeostasis in dairy cows. Since the incidence of peripartal hypocalcemia and the reported beneficial effects of the treatment are both associated with the lactation number, we investigated pharmacokinetic aspects of 25-OHD3 related to the age of dairy cows. The daily oral administration of 3mg 25-OHD3 in rapeseed oil as well as a treatment with 4 and 6mg included in the feed during the last eight to ten days of gestation resulted in linear dosage- and age-dependent increases in plasma 25-OHD3. After parturition the administration was stopped and blood samples were taken to calculate the plasma half-life. Irrespective of the supplemented dosage, cows starting the 2nd lactation showed a significantly longer plasma half-life of 25-OHD3 than cows starting the 3rd or higher lactation. Age-dependent differences in the increase of plasma 25-OHD3 could already be found before parturition when calcium homeostasis was not yet significantly challenged. Additionally, no correlations between plasma half-life of 25-OHD3 and 1,25-dihydroxyvitamin D3, PTH or the bone resorption marker CrossLaps were observed after parturition. Thus we conclude that the influence of the lactation number on the pharmacokinetics of 25-OHD3 is related directly to the age of the cows. This article is part of a Special Issue entitled 'Vitamin D Workshop'.