Discrepancies of bovine haptoglobin concentrations between serum and plasma using two different anticoagulants and a colorimetric assay based on peroxidase activity.

BACKGROUND: Haptoglobin (Hp) is an emerging diagnostic marker in cattle, and knowledge of suitable sample types and measurement methods is important. OBJECTIVES: The aims of this study were to compare the results of a colorimetric assay (CA) and an ELISA for bovine Hp using serum, EDTA plasma, and lithium-heparinized (LH) plasma, respectively, and to assess the diagnostic potential for puerperal metritis. METHODS: In experiment 1, Hp was measured in pooled aliquots of serum (n = 10), EDTA plasma (n = 10), and LH plasma (n = 10) of 100 healthy fresh lactating dairy cows from 10 farms using both the CA and the ELISA. In experiment 2, five healthy and five cows with acute puerperal metritis were sampled, and Hp was determined using both assays for all three sample types. In experiment 3, aliquots of serum and LH plasma from cows in different lactation stages were transferred into plain, EDTA-coated, and LH-coated tubes and mixed before colorimetric analyses. Distilled water was also placed into each tube type and treated similarly. RESULTS: Plasma samples measured with the CA showed on average 2.3 (EDTA) and 2.5 (LH) times higher Hp concentrations compared with serum, whereas no differences were seen with the ELISA results between sample types. Based on a clinical cut-off value, both methods differentiated sick from healthy cows. Haptoglobin measurements with the ELISA were less precise compared with CA measurements due to high dilutions. No influence of the anticoagulants on the CA was observed. CONCLUSIONS: Due to measurement discrepancies between serum and plasma, CAs for bovine Hp based on peroxidase activity should be performed with serum, or specific reference ranges for plasma samples should be established. In this study, CA results obtained with LH plasma were more precise than results obtained with EDTA plasma. Both the CA and the ELISA are suitable diagnostic tools for the diagnosis of puerperal metritis, but CA measurements were more precise in this study.

Effect of 2 dosages of prepartum cholecalciferol injection on blood minerals, vitamin D metabolites, and milk production in multiparous dairy cows: A randomized clinical trial.

The objective of the present study was to evaluate the effect of 2 dosages of prepartum cholecalciferol injection on blood minerals, vitamin D metabolites, and milk production. Cows entering their second or greater lactation (n = 158) were randomly assigned to a control group (CON) or one of 2 treatment groups receiving either 6 × 106 IU (6VitD) or 12 × 106 IU (12VitD) cholecalciferol intramuscularly on d 275 ± 1.2 (SD) of gestation. Concentrations of serum total Ca (tCa), phosphate, and Mg were determined on 1, 2, 3, 5, 7, and 10 d in milk (DIM). For a subsample of 30 cows entering the third lactation (n = 10/group), these samples were analyzed for cholecalciferol, 25-hydroxycholecalciferol (25-OHD3), and 24,25-dihydroxycholecalciferol (24,25-[OH]2D3). In these cows, we also determined 1,25-dihydroxycholecalciferol (1,25-[OH]2D3), the biologically most active metabolite, on 1, 2, 3, and 5 DIM. Repeated measures ANOVA was performed to evaluate the effect of different dosages of cholecalciferol on blood minerals, vitamin D metabolites, and milk yield over the first 5 test days after calving. Binary outcomes such as retained placenta and metritis were analyzed using a chi-squared test. Although the 12VitD treatment increased tCa concentrations on 1, 2, and 3 DIM compared with CON, administration of 6VitD increased tCa concentrations only on 1 DIM. Compared with CON cows and 6VitD cows, 12VitD cows had greater serum phosphate concentration during the first 10 DIM. Furthermore, 6VitD cows had greater serum phosphate concentrations compared with CON cows. On the contrary, 12VitD cows had lower serum Mg concentrations during the first 10 DIM compared with CON and 6VitD cows. Cholecalciferol was increased by the treatment and decreased quickly until 10 DIM. In respect to 25-OHD3, the 6VitD treatment resulted in a 4.1-fold increase in comparison to the CON group, while a 6.5-fold increase was observed in 12VitD animals. The vitamin D metabolite 24,25-(OH)2D3 increased linearly with 25-OHD3 serum levels, resulting in the highest concentrations in the 12VitD group. An increase of 1,25-(OH)2D3 until 3 DIM was observed in all cows. However, this rise was most pronounced in the CON group. The incidence of retained placenta was 1.9%, 11.5%, and 29.6%, and that of metritis was 11.5%, 15.4%, and 31.5% for CON, 6VitD, and 12VitD cows, respectively. Although none of the treated cows exerted clinical signs of hypocalcemia, one cow in CON incurred clinical hypocalcemia. Cows of the 12VitD group had a lower milk yield over the first 5 monthly test days compared with the control and 6VitD group (42.2 ± 0.5, 42.0, ± 0.5 and 40.7 ± 0.5 kg for control cows, 6VitD cows and 12VitD cows, respectively). Although no negative side effects were observed in 6VitD cows, we do not recommend the general application of 6 × 106 IU cholecalciferol before calving as positive effects on calcium homeostasis were marginal and restricted to the first DIM. The present findings confirm that the application of 12 × 106 IU cholecalciferol negatively affected milk production on this farm.

Effects of a single transdermal administration of flunixin meglumine in early postpartum Holstein Friesian dairy cows: Part 1. Inflammatory and metabolic markers, uterine health, and indicators of pain.

The objectives of this study were to assess the effects of a single transdermal administration of flunixin meglumine (FM) in early postpartum Holstein Friesian dairy cows on serum concentrations of inflammatory and metabolic markers, uterine health, and indicators of pain. The hypothesis was that the anti-inflammatory, antipyretic, and analgetic effects of the pharmaceutic agent would reduce systemic inflammation, resulting in improved metabolic and inflammatory profile, diminished incidence of metritis, and reduced expression of pain. A total of 500 cows (153 primiparous, 347 multiparous) from 3 different commercial dairy farms in the northeast of Germany were included in a randomized controlled clinical trial. Farms were preselected based on high haptoglobin concentrations in their fresh lactating cows. Cows were excluded if they had experienced dystocia, stillbirth, or twin birth, or if they showed any signs of milk fever, retained fetal membranes, or fever (>40°C). The cows were treated once with either FM (3.33 mg/kg) or a placebo as control (CON) through transdermal administration between 24 to 36 h postpartum (d 2). General health examinations were performed (daily from d 2-8 and additionally on d 15 postpartum), vaginal discharge was assessed using the Metricheck device (d 8 and 15 postpartum) and serum samples were analyzed for inflammatory and metabolic markers (d 2, 4, and 6 postpartum). Effects of treatment, parity, sampling day, and their interactions were evaluated using mixed effects models. Primiparous cows treated with FM showed lower serum haptoglobin concentrations (0.90 ± 0.08 vs. 1.17 ± 0.07 g/L; ± standard error of the mean) and higher serum albumin concentrations (35.5 ± 0.31 vs. 34.8 ± 0.31 g/L) on d 6 postpartum. They also had a lower risk for purulent vaginal discharge with or without a fever compared with CON cows on d 15 postpartum (odds ratio for CON vs. FM: 1.63, 95% CI: 1.26-2.00), and body temperature was lower throughout the first 15 d in milk (39.1 ± 0.11 vs. 39.2 ± 0.11°C). Multiparous cows treated with FM had lower serum ß-hydroxybutyrate concentrations on d 4 postpartum (0.71 ± 0.05 vs. 0.78 ± 0.05 mmol/L) and d 6 postpartum (0.74 ± 0.05 vs. 0.80 ± 0.05 mmol/L). Regardless of parity, FM-treated cows were significantly less likely to abduct their tail from their body (14.3 vs. 23.6%) and show an arched back (27.9 vs. 39.7%) on the day after treatment compared with CON cows. It can be concluded that FM treatment slightly reduced inflammation and diminished the risk for metritis in primiparous cows, improved metabolic profile in multiparous cows, and reduced expressions of pain in all cows.

Effects of a single transdermal administration of flunixin meglumine in early postpartum Holstein Friesian dairy cows: Part 2. Milk yield, culling risk, and reproductive performance.

This study was conducted to assess the effects of a single transdermal administration of flunixin meglumine (FM) in early postpartum Holstein Friesian dairy cows on milk yield, culling risk, and reproductive performance. We hypothesized that FM treatment would reduce systemic inflammation, leading to higher milk yield, reduced culling risk, and better reproductive performance in the subsequent lactation. Holstein Friesian dairy cows [n = 500, 153 primiparous (PRIM), 347 multiparous (MULT)] from 3 farms in northeast Germany were enrolled in a prospective, randomized controlled clinical trial. Farms at risk for cows with excessive postpartum inflammation were identified in a preliminary trial by measuring serum haptoglobin concentrations in their fresh lactating cows. Only cows that had a eutocic birth and delivered a singleton calf alive, with no signs of milk fever or retained fetal membranes and rectal temperature ≤40°C at first clinical examination, were included within 24 to 36 h postpartum. Treatment included a single transdermal administration of either FM (3.33 mg/kg) or a placebo as control (CON). Milk production, milk solids, urea, and somatic cell count were recorded monthly for 8 mo after calving. Culling risk, first-service conception risk, and days open were retrieved from the farms' herd management software. Separate models for PRIM and MULT cows were built for most parameters because of significant effects of parity and parity × treatment interaction. Energy-corrected milk yield from 8 monthly Dairy Herd Improvement-equivalent tests was slightly greater in PRIM cows treated with FM (29.51 and 30.73 ± 1.35 kg, CON vs. FM), whereas it was reduced in treated MULT cows (38.23 and 37.47 ± 1.17 kg, CON vs. FM) compared with CON. Milk fat and protein yields were greater in FM-treated PRIM cows and lower in treated MULT cows compared with CON. Milk urea and somatic cell count were not affected by treatment. No differences in culling risk, first-service conception risk, or days open were observed. We conclude that a single transdermal administration of FM in early postpartum dairy cows on farms at risk for excessive postpartum inflammation slightly increased milk, milk fat, and milk protein yields in PRIM cows and decreased these variables in MULT cows. Neither culling risk nor fertility was affected by treatment in this study.

[Analyses of the relationship between the concentrations of essential trace elements in total mixed ration and faeces samples from Holstein Friesian dairy cows and the estimation of faeces reference values]. / Untersuchungen zum Zusammenhang der Konzentrationen an essenziellen Spurenelementen in totalen Misch­rationen- und Kotproben bei Holstein Friesian Milchkühen und Ableitung von Kotreferenzwerten.

INTRODUCTION: Undersupply and oversupply of dairy cows with essential trace elements should be avoided from the point of view of animal health and environmental pollution. The aim of the study was to proof the results faecal analyzes on the nutritional supply situation with essential trace elements in comparison to measurements from total mixed ration (TMR) analyses. Concentrations of iron (Fe), copper (Cu), zinc (Zn), Mangan (Mn), selenium (Se), cobalt (Co), molybdenum (Mo) were measured as representative multicentric random sample in 574 TMR, 600 pooled faecal and blood samples from dairy cows. Trace elements, Fe in 0 %, Se 18 %, Zn 43 %, Cu 52 %, Co 53 % and Mn 59 %, were within the target range. Undersupply was found in 0 to 8 % and an oversupply in 39 to 100 % of the TMR samples. The tolerance limit for Fe was exceeded in 11 % and for Mo in 13 %. The results indicate the need for optimized ration design. The trace element concentrations in the faecal samples varied over a wide range. Results of pooled faecal samples (n=10 per pool) corresponded closely with the mean values calculated from the individual samples (r2 > 0,9). Correlation between element concentrations in the TMR and faecal samples for Fe (r=0,687), Cu (r=0,675), Zn (r=0,635), Mn (r=0,656), Se (r=0,573), Co (r=0,795), and Mo (r=0,708) were highly significant. No correlations were detected between the element concentrations in the TMR and in the blood samples. The regression analyzes showed a highly significant linear adjustment of the measured value distribution around the regression lines over the entire value range including the zero point. Reference values were calculated for the faecal concentrations of Cu, Zn, Mn, Se and Co. Tolerance reference limits in the TMR were calculated for the faecal concentrations of Fe and Mo. The trace element concentrations in the TMR samples reflect the nutritional supply situation. Knowledge of the nutritional supply situation of the herd is of great importance for the stock management of dairy cow. Fecal sample analyzes can be used as an alternative to TMR examinations to assess the nutritional supply situation.

INTRODUCTION: Les sous-approvisionnements et les surapprovisionnements des vaches laitières en oligo-éléments essentiels doivent être évités du point de vue de la santé animale et de la pollution de l'environnement. L'objectif de cette étude était de vérifier la pertinence des analyses de fèces pour évaluer l'apport nutritionnel en oligo-éléments essentiels en les comparant aux valeurs mesurées dans les analyses de RTM (ration totale mélangée). Dans un échantillon aléatoire multicentrique représentatif, les concentrations de fer (Fe), de cuivre (Cu), de zinc (Zn), de manganèse (Mn), de sélénium (Se), de cobalt (Co) et de molybdène (Mo) ont été mesurées dans 574 échantillons de RTM et 600 échantillons de fèces regroupés de vaches laitières et des échantillons de sang ont été analysés. La plage cible pour la teneur en oligo-éléments dans la RTM a été respectée pour Fe à 0 %, Se à 18 %, Zn à 43 %, Cu à 52 %, Co à 53 % et Mn à 59 %. La plage de sous-alimentation a été constatée dans 0 à 8 % des échantillons de RTM, la plage de suralimentation dans 39 à 100 %. La limite de tolérance pour le Fe a été dépassée dans 11 %, pour le Mo dans 13 %. Ces résultats indiquent la nécessité d'optimiser les rations. Dans les échantillons de fèces, les concentrations en oligo-éléments variaient sur une large plage. Les résultats d'analyse des échantillons de fèces regroupés (n=10 par pool) correspondaient étroitement, avec une valeur r2 supérieure à 0,9, aux valeurs moyennes calculées à partir des échantillons individuels. Il y avait des corrélations très significatives entre les concentrations d'éléments dans les échantillons de RTM et de fèces pour Fe (r=0,687), Cu (r=0,675), Zn (r=0,635), Mn (r=0,656), Se (r=0,573), Co (r=0,795), et Mo (r=0,708). Aucune corrélation n'a pu être établie entre les concentrations d'éléments dans le RTM et dans les échantillons de sang. Les analyses de régression ont montré un ajustement linéaire hautement significatif de la distribution des valeurs mesurées autour de la droite de régression sur l'ensemble de la plage de valeurs, y compris le point zéro. Des valeurs de référence ont été calculées pour les concentrations de Cu, Zn, Mn, Se et Co dans les fèces. Pour les concentrations de Fe et Mo dans les fèces, des limites de référence ont été calculées pour les limites de tolérance dans la RTM. Les concentrations en oligo-éléments dans les échantillons de RTM reflètent la situation nutritionnelle. Pour le suivi des troupeaux de vaches laitières, il est très important de connaître l'état nutritionnel du troupeau. Les analyses d'échantillons de fèces peuvent être utilisées comme alternative aux analyses de RTM pour évaluer la situation nutritionnelle.

Randomized clinical trial to evaluate the effects of a prepartum cholecalciferol injection on postpartum serum calcium dynamics and health and performance in early-lactation multiparous dairy cows.

The objectives of the present study were (1) to evaluate the effect of prepartum cholecalciferol treatment on serum Ca concentration during the first 10 d after calving and (2) to evaluate the effect of treatment on subsequent health and performance. Multiparous Holstein cows (n = 377) from one dairy farm were fed a negative dietary cation-anion difference diet (-31 mEq/kg of DM) for the last 21 d of gestation. On d 275, the animals were randomly assigned to a control or a treatment group. Cows in the control group were left untreated, and cows in the treatment group received an injection of 12 × 106 IU of cholecalciferol intramuscularly on the day of enrollment. If treated cows did not deliver the calf within 6 d, they were reinjected with 10 × 106 IU of cholecalciferol. Blood samples were drawn on 1, 2, 3, 5, 7, and 10 days in milk (DIM) and analyzed for serum Ca, P, and Mg concentrations. In a subsample of cows (50 control cows, 35 cows treated once with cholecalciferol, and 15 cows treated twice) serum haptoglobin, nonesterified fatty acids, ß-hydroxybutyrate, and 25-hydroxycholecalciferol concentrations were analyzed on 1, 5, and 10 DIM. Binary data [retained placenta (RP), metritis] were analyzed using logistic regression models. Repeated measures ANOVA with first-order autoregressive covariance was performed to evaluate the treatment effect on milk yield over the first 10 test days after parturition, 25-hydroxycholecalciferol, serum Ca, P, Mg, ß-hydroxybutyrate, nonesterified fatty acids, and haptoglobin concentrations. Cox proportional hazards were used to model the time to event outcomes (time to pregnancy within 200 d, culling until 300 DIM). After enrollment of 31.4% of cows and a preliminary analysis, adverse reactions became apparent, and the study was stopped. Cows treated with cholecalciferol had a greater risk of incurring RP and metritis. The adjusted mean incidences were 2.0%, 7.7%, and 4.0% for RP, and 21.6%, 39.3%, and 33.3% for metritis for control cows, cows treated once, and cows treated twice with cholecalciferol, respectively. Compared with control cows, cows injected once with 12 × 106 IU of cholecalciferol produced less energy-corrected milk on the first (-3.76 kg) and second (-2.75 kg) test days, respectively. Cows injected twice with cholecalciferol (12 × 106 IU of cholecalciferol and 10 × 106 IU 1 wk later) had a reduced milk yield only at first test day (-3.80 kg). Treatment with cholecalciferol led to a significant increase in 25-hydroxycholecalciferol on d 1, 5, and 10 after calving. Serum Ca and P concentrations were significantly increased in cows treated with cholecalciferol, but serum Mg concentrations were significantly reduced. Haptoglobin concentrations were significantly increased on 5 DIM in cows injected once with 12 × 106 IU of cholecalciferol. Although we observed no effect of treatment on culling until 300 DIM, time to pregnancy was delayed by 34 d in cows injected once with 12 × 106 IU of cholecalciferol. In the present study, injection with 12 × 106 IU of cholecalciferol had detrimental effects on health and milk production despite the beneficial effects on Ca homeostasis.

Association between serum calcium dynamics around parturition and common postpartum diseases in dairy cows.

The objective of this study was to compare periparturient serum Ca dynamics (CaDyn) in cows with and without diseases in early lactation. The study enrolled 1,949 cows from a commercial dairy farm in northern Germany. Blood samples were drawn 7 d before expected calving date and on d 0, 1, 3, and 7 after calving and analyzed for serum Ca concentration. Cows were monitored for clinical hypocalcemia (CH), ketosis, left displaced abomasum (LDA), retained placenta, acute puerperal metritis (APM), mastitis, and pneumonia. To evaluate the association between CaDyn and diseases during the transition period, repeated measures ANOVA with first-order autoregressive covariance were performed. Serum CaDyn of healthy cows (i.e., without any of the aforementioned diseases) was compared with CaDyn of cows with one of the aforementioned diseases (CH, ketosis, APM, mastitis, LDA, and pneumonia), and cows with multiple diseases (CH+, ketosis+, APM+, mastitis+, LDA+, and pneumonia+). Separate models were built for primiparous and multiparous cows. For primiparous cows, we evaluated the association between CaDyn and ketosis (healthy cows vs. cows with ketosis vs. cows with ketosis+) and CaDyn and APM (healthy cows vs. cows with APM vs. cows with APM+). The same models were built for multiparous cows. Three additional models were built for multiparous cows to evaluate the association between CaDyn and CH (healthy cows vs. cows with CH vs. cows with CH+), mastitis (healthy cows vs. cows with mastitis vs. cows with mastitis+), or LDA (healthy cows vs. cows with LDA vs. cows with LDA+). In primiparous cows, serum Ca concentrations of cows with ketosis, APM, and APM+ were significantly reduced on d 3 and 7 after calving, compared with healthy cows. Serum Ca concentrations of primiparous cows with ketosis+ were reduced on d 3, but not on d 7 after calving. Multiparous cows with CH had significantly reduced serum Ca concentrations on d 0, 1, and 3 compared with healthy cows. On d 3 and 7, serum Ca concentration of CH+ cows was significantly reduced compared with healthy multiparous cows. Multiparous cows with ketosis and ketosis+ had significantly reduced serum Ca concentrations on d 1 and 3 compared with healthy cows. Cows with APM+ had significantly increased serum Ca concentrations on d 0 and reduced serum Ca concentrations on d 3, compared with healthy cows. Whereas multiparous cows with mastitis had a reduced serum Ca concentration on d 1, mastitis+ cows had a reduced serum Ca concentration on d 1 and 3, compared with healthy multiparous cows. Overall, multiparous cows with LDA+ had reduced serum Ca concentrations. Especially a delayed onset of hypocalcemia (d 3 and 7) was indicative for the development of disease in primiparous cows. In multiparous cows, reduced serum Ca concentrations on d 1 and 3 were associated with occurrence of diseases. Future studies should evaluate whether reduced serum Ca concentrations are a cause or concomitant circumstance of diseases in early lactation.

Serum calcium dynamics within the first 3 days in milk and the associated risk of acute puerperal metritis.

The objectives of this study were to evaluate postpartum serum calcium dynamics for different parity groups of dairy cows and to assess whether serum calcium concentration on d 0, 1, and 3 postpartum was associated with the risk of developing acute puerperal metritis (APM). The study took place on a commercial dairy farm in northern Germany and included 4,043 Holstein dairy cows. Calving difficulties, such as dystocia, twins, or stillbirth, were recorded. Blood samples were obtained on d 0, 1, and 3 after calving for analysis of serum calcium concentration. Animals were examined daily for clinical symptoms of retained placenta, APM, mastitis, and displaced abomasum until 10 d in milk. To determine serum calcium dynamics postpartum, we performed repeated-measures ANOVA with first-order autoregressive covariance. A logistic regression model was used to evaluate the association of serum calcium concentration with the risk of developing metritis. Serum calcium concentration was affected by time relative to calving, parity, and APM. Increasing parity negatively affected serum calcium concentration on d 0 and 1. Serum calcium concentration reached its lowest level on d 1 and 3 in multiparous and primiparous cows, respectively. The concentration increased from d 1 to 3 in multiparous cows and decreased from d 0 to 3 in primiparous cows. The association of APM and serum calcium dynamics varied by parity. On d 3, serum calcium concentration was significantly lower in animals with subsequent APM than in those without APM. The overall incidence of APM was 12.0% (primiparous cows, 20.4%; multiparous cows, 8.6%). An association existed between serum calcium concentration on d 3 after calving and APM. Primiparous cows had an odds ratio of 0.12 for serum calcium concentration on d 3, indicating that a primiparous cow with serum calcium concentration of 2.5 mmol/L had a 88% lower chance of developing APM compared with a cow with a concentration of 1.5 mmol/L. Multiparous cows had an odds ratio of 0.34 for serum calcium concentration on d 3, indicating that a multiparous cow with serum calcium concentration of 2.5 mmol/L had a 66% lower chance of developing APM compared with a cow with a concentration of 1.5 mmol/L. Primiparous cows with low serum calcium concentration had the highest predicted probability of developing APM. Our results reveal a dynamic in serum calcium concentration in the first 3 d in milk. Consequently, the day of sampling and the observed risk period for hypocalcemia are important when conducting epidemiological studies to evaluate associations between hypocalcemia and clinical diseases.

[Evaluation of sulfur status in dairy cows in Germany]. / Untersuchungen zur Schwefelversorgung von Milchkühen in Deutschland.

OBJECTIVE: Sulfur deficiency and toxicity in livestock is associated with decreased performance and diseases. However, little is known about the diagnostics of sulfur status in dairy herds. The objectives of this study were to describe sulfur concentrations in feed rations in German dairy herds, to assess associations between feed sulfur concentrations and health and milk yield, to determine suitable laboratory diagnostic methods of sulfur status, and to propose reference values for the assessment of sulfur status in different sample media. MATERIALS AND METHODS: Between 2006 and 2014, a total of 569 dairy herds were visited. Blood, urine, and hair samples were obtained from 10 animals per group in the groups -3 to 0, 0 to 1, 3 to 5, and 15 to 18 weeks post partum (p. p.) (farms > 200 animals) or -3 to 0, 0 to 5, and 6 to 20 weeks p. p. (farms ≤ 200 animals). Pooled whole blood, serum, plasma, urine, and hair samples (n = 5663) were analyzed by ICP-OES. The sulfur content of the total mixed ration (TMR) of the group -3 to 0 weeks p. p. was determined in a certified feed laboratory (n = 625). Data analyses were performed using ANOVA, Chi-squared test and Spearman-correlation. Reference values were determined using the 2.5% and 97.5% percentiles. RESULTS: Median sulfur concentration in the feed rations was 2.3 g/kg dry matter (DM). Deficiency (< 1.6 g/kg DM; 31% of farms) and oversupply (> 4.0 g/kg DM; 11% of farms) of sulfur in feed rations were regularly diagnosed. Sulfur deficiency was associated with reduced milk yield (p  <   0.001), retained placenta (OR  =   1.74; p  =  0.037), milk fever (OR  =  2.68; p  <  0.001) and silent heat (OR  = 2.56; p   =  0.014). Positive correlations were found between sulfur concentration in feed rations and sulfur content in urine (r  =  0.50), serum (r  =  0.17), plasma (r  =  0.18) and whole blood (r = 0.16). Reference values for sulfur diagnostics in dairy herds were proposed. CONCLUSION AND CLINICAL RELEVANCE: Diagnostics of sulfur status in dairy herds is relevant for veterinary practitioners and can be performed using TMR analysis and clinical laboratory parameters. Urine can be considered a short-term parameter, serum, plasma, and whole blood as intermediate parameters, and hair as a long-term parameter for the evaluation of the sulfur status.

[Milk yield and environmental factors: Multiple regression analysis of the association between milk yield and udder health, fertility data and replacement rate]. / Milchleistung und Umweltfaktoren. Multiple Regressionsanalyse über die Beziehungen der Milchleistungshöhe zur Eutergesundheit, Fruchtbarkeit und Remontierungsrate.

OBJECTIVE: The relationship between milk yield and both fertility and general animal health in dairy herds is discussed from opposing viewpoints. The hypothesis (1) that raising the herd milk yield would decrease fertility results, the number of milk cells as an indicator for udder health and the replacement rate as a global indicator for animal health as well as increasing the occurrence of specific diseases as a herd problem was compared to the opposing hypotheses that there is no relationship (2) or that there is a differentiated and changing relationship (3). MATERIAL AND METHODS: A total of 743 herd examinations, considered independent, were performed in 489 herds between 1995 and 2010. The milk yield, fertility rate, milk cell count, replacement rate, categorized herd problems and management information were recorded. The relationship between the milk yield and both the fertility data and animal health was evaluated using simple and multiple regression analyses. RESULTS: The period between calving and the first service displayed no significant relationship to the herd milk yield. Simple regression analysis showed that the period between calving and gestation, the calving interval and the insemination number were significantly positively associated with the herd milk yield. This positive correlation was lost in multiple regression analysis. The milk cell count and replacement rate using both the simple and multiple regression analyses displayed a significant negative relationship to the milk yield. CONCLUSION AND CLINICAL RELEVANCE: The alternative hypothesis (3) was confirmed. A higher milk yield has no negative influence on the milk cell count and the replacement rate in terms of the udder and general health. When parameterizing the fertility, the herd milk yield should be considered. Extending the resting time may increase the milk yield while preventing a decline in the insemination index.

Relationship between body condition score and ultrasound measurement of backfat thickness in multiparous Holstein dairy cows at different production phases.

BACKGROUND: The validity of the body condition score (BCS) for evaluating subcutaneous fat has been questioned because of its subjectivity. With the advent of ultrasonography, backfat thickness (BFT) can be objectively measured and used to assess the energy status of dairy cows in conjunction with BCS. OBJECTIVE: Determine the relationship between BCS and ultrasound measurements of BFT throughout the lactation cycle. METHODS: Cows (n = 1123) were selected randomly and classified according to their lactation stage into four different groups. BCS was determined using a 5-point scale and BFT measurements were obtained using a 5-MHz linear transducer. RESULTS: The correlation coefficient (r) for the relationship between BCS and BFT varied according to the stage of lactation. There was a high correlation (r = 0.96-0.98) and relationship (r(2) = 0.92-0.96) between BCS and BFT. The highest correlation was found at BFT values of 10-35 mm and BCS of 2-4.5 units. CONCLUSION: BCS is a practical tool for evaluating body condition; furthermore, it may be as valid as BFT for the estimation of subcutaneous fat in Holstein dairy cows over time.

Alkalinizing effect of NaHCO3 with and without glucose when administered orally to euhydrated neonatal dairy calves.

The use of oral rehydration solutions (ORS) is well established as an effective treatment to correct water-, electrolyte-, and acid-base balance in diarrheic calves. The main ingredients of a commercial ORS are Na, glucose, and alkalinizing agents, such as NaHCO3. Particular importance is attributed to the combination of glucose and Na at a specific ratio to optimize intestinal sodium, and thereby water uptake, through the sodium-glucose co-transport. Enhancing intestinal Na absorption by combining glucose and Na in an ORS has the potential to improve the alkalinizing effect of an ORS according the strong ion theory. The objective of this study was to investigate the effect of glucose on the alkalinizing effect of NaHCO3 when administered orally. Nine healthy neonatal Holstein-Friesian calves underwent 3 oral treatments with 2-L solutions of NaHCO3 (150 mmol/L), glucose (300 mmol/L), and glucose + NaHCO3 (300 mmol/L + 150 mmol/L, respectively) in randomized order. Arterial and venous blood was obtained before treatment and in 30-min intervals thereafter for blood gas analysis and determination of plasma protein and electrolyte concentrations. Urine was collected volumetrically to determine urine volume, osmolality, pH, net acid excretion, and renal Na excretion after treatment. Plasma volume changes were extrapolated from plasma protein concentration changes. Treatment and time effects were tested with repeated measures ANOVA. Only subtle differences between oral administration of NaHCO3, with and without glucose, were observed for the change of the standard HCO3 concentration relative to baseline. No differences in plasma Na, plasma volume expansion, renal Na, net base excretion, urine volume, or pH could be identified between animals treated orally with NaHCO3 with and without glucose. Similarly, no differences in blood glucose concentration, plasma volume expansion, urine volume, or renal glucose excretion were observed in the 8h after treatment when comparing oral glucose treatment with and without NaHCO3. Our results indicate that combination of NaHCO3 with glucose in a hypertonic ORS only had a minor effect on the alkalinizing effect of NaHCO3, which is unlikely to be of clinical relevance. The combination of NaHCO3 and glucose neither improved Na, glucose, nor water absorption in euhydrated neonatal dairy calves, questioning the relevance of a specific ratio between Na and glucose in ORS for calves.

Variations in copper concentration and ceruloplasmin activity of dairy cows in relation to lactation stages with regard to ceruloplasmin to copper ratios.

Variations of copper (Cu) concentration and ceruloplasmin (Cp) activity in serum and plasma of dairy cows at different stages of lactation were assessed in 240 Holstein dairy cows. Furthermore, ceruloplasmin to copper ratios were also investigated. The cows were classified according to their lactation stages into four different groups as close-up stage (3-1 week(s) antepartum), fresh lactation stage (0-1 week postpartum), early lactation stage (3-5 weeks postpartum), and mid-lactation stage (15-18 weeks postpartum). Each group consisted of 60 multiparous cows. Serum and heparinised plasma samples were obtained from each cow. Concentrations of serum copper (sCu) and plasma copper (pCu) as well as activities of plasma Cp (pCp) were higher in the group of fresh lactation stage than other groups (P < 0.05). Serum Cp (sCp) exhibited no significant difference between fresh lactation and early lactation groups (P > 0.05). Low correlations were obtained between sCp and pCu, sCp and sCu, and sCu and pCu. Plasma copper concentration and plasma ceruloplasmin activity were higher than serum copper concentration and serum ceruloplasmin activity, respectively (P < 0.001). The ratios of Cp activity to Cu concentration (Cp/Cu) were not significantly changed in the different lactation stages of dairy cows (P > 0.05). Use of sCp/pCu and sCp/sCu rather than pCp/pCu will reduce the calculated value of Cp/Cu. Furthermore, for evaluation of copper status, use of sCp/sCu or sCp/pCu identified more animals as 'low' and 'marginal' than using pCp/pCu (P < 0.001). It can be concluded that ceruloplasmin and copper undergo a physiological increase just after calving; thus, their values should be interpreted with caution during assessment of copper status. Plasma measurements should be used for calculation of Cp/Cu, and further research is required to refine diagnosis criteria for use of such ratio in determining copper status in dairy cows.

Plasma potassium-lowering effect of oral glucose, sodium bicarbonate, and the combination thereof in healthy neonatal dairy calves.

Hyperkalemia is a common complication in neonatal diarrheic dairy calves and is corrected by administration of glucose or sodium bicarbonate. Although the hypokalemic effect of glucose is well established in other species, controversial results are reported for sodium bicarbonate. Our objective was to study the effect and mechanism of action of glucose and sodium bicarbonate on the potassium homeostasis of healthy neonatal dairy calves. Nine healthy neonatal Holstein-Friesian calves underwent 3 oral treatments with 2L of NaHCO(3) (150 mmol/L), glucose (300 mmol/L), and glucose+NaHCO(3) solution (300 mmol/L+150 mmol/L) in randomized order. Blood was obtained before treatment (T(0)) and at 30-min intervals thereafter. Changes between each time point and T(0) were determined for all parameters. Urine was collected volumetrically to determine total renal potassium excretion over an 8-h posttreatment period. Plasma volume changes were extrapolated from changes in plasma protein concentration. Treatment and time effects were tested with repeated-measures ANOVA. Multivariate stepwise regression analysis using dummy variable coding was conducted to identify associations between changes in plasma potassium concentration ([K]) and changes in plasma glucose concentration ([glucose]), blood base excess, and plasma volume. Oral glucose and sodium bicarbonate treatments decreased [K] by 25 and 19%, respectively, whereas the combination of both compounds caused an intermediate [K] decline (22%). For the glucose treatment, the decline in [K] was only associated with changes in plasma [glucose] (partial R(2)=0.19). In NaHCO(3)-treated calves, [K] decline was associated with change of extracellular volume (partial R(2)=0.31) and blood base excess (partial R(2)=0.19). When glucose and NaHCO(3) were combined, [K] decline was associated with changes in plasma volume (partial R(2)=0.30), BE (R(2)=0.22), and [glucose] (partial R(2)=0.03). Our results indicate that glucose lowers plasma [K] mainly through an insulin-dependent intracellular translocation of K, whereas NaHCO(3) causes hypokalemia through hemodilution followed by intracellular translocation of K caused by the strong ion effect. The combination of glucose and NaHCO(3) at the dosage used in this study does not have an additive hypokalemic effect. When combined, hemodilution and strong ion effect have the strongest effect on plasma [K], whereas the insulin-dependent effect of glucose appears to be blunted.

Comparison of the impact of different anionic salts on the acid-base status and calcium metabolism in non-lactating, non-pregnant dairy cows.

Eleven mature, non-lactating, non-pregnant, Holstein-Friesian crossbred cows were given, via a ruminal cannula, 2000mEq of one of three chloride salts, four sulfate salts, two combinations of anionic salts (AS), sodium chloride (as neutral salt), or water as control. The salts and controls were assigned in an 11x11 Latin square and the cows were randomly distributed. All of the AS induced a metabolic acidosis that resulted in a small reduction of blood pH, base excess, and bicarbonate (P<0.001), and notable changes in urinary pH, net acid base excretion (P<0.001), and urinary calcium excretion (P<0.001). Only calcium chloride had a significantly greater impact on acid-base status (ABS) than the sulfate salts. The effect of other chloride salts did not differ from calcium sulfate. There was no indication that chloride salts in general have a greater impact than sulfate salts on the ABS.

The effect of various anionic salts on ruminant pH and short-chain fatty acids in non-pregnant and non-lactating cows.

AIM: To evaluate whether the different anionic salts used in the prevention of parturient paresis have an impact on the ruminal pH and the production of short-chain fatty acids (SCFA) in mature non-pregnant, non-lactating cows. METHODS: Eleven Holstein-Friesian crossbred cows were administered 2,000 mEq of either one of three chloride salts, viz CaCl2, MgCl2, or NH4Cl; four sulphate salts, viz CaSO4, CaSO4 with a grain size of 10 microm, MgSO4, or (NH4)2SO4; two combinations of anionic salts, viz CaCl2+MgSO4, or CaSO4+NH4Cl; NaCl; or water, via a ruminal cannula over a 14-day treatment period. The salts and controls were assigned in an 11 x 11 Latin square, and the cows were distributed randomly. Ruminal fluid was collected four times in each treatment period for monitoring the ruminal pH, and four times a day at Days 7 and 14 for monitoring any changes in the concentrations of SCFA. RESULTS: Feeding anionic salts did not change the ruminal pH, total concentration of SCFA, or distribution pattern of the main SCFA acetic acid, propionic acid, butyric acid or valeric acid (p>0.05). CONCLUSIONS AND CLINICAL RELEVANCE: Feeding anionic salts has no negative side effect on the ruminal pH and concentrations of SCFA in mature non-pregnant, non-lactating cattle. Impaired function of the rumen due to the feeding of anionic salts is not likely.

Liver phosphorus content in Holstein-Friesian cows during the transition period.

Hepatic lipidosis and hypophosphatemia are frequently observed in high-yielding periparturient dairy cows. Objectives of this study were to investigate the association of the liver P content with the degree of liver fat accumulation and serum P concentration and to characterize the change in liver P content throughout the transition period. In a cross-sectional study, liver biopsies obtained from 33 Holstein-Friesian cows 14 d postpartum (p.p.) were assayed for total lipid (TLip), triacylglycerol, DNA, P, Mg, K, Na, and Ca content. Serum samples obtained at the time of biopsy were analyzed for indices of liver function and injury and the serum P concentration was determined. From this cross-sectional study, 6 cows were selected for a longitudinal study and liver tissue obtained from the 6 cows on d -65, -30, -14, 1, 14, 28, and 49 relative to calving was assayed. The amounts of P, K, Mg, Na, and Ca were expressed as amount in dry weight (DW), wet weight (WW), nonfat wet weight (NFWW), and indexed to DNA. In the cross-sectional study, P(DW) and P(WW) decreased with increasing TLip, whereas P(NFWW) and P(DNA) were independent of TLip. Values for P(DNA) varied widely, whereas P(NFWW) varied within a narrow range. Stepwise regression analysis revealed the strongest associations between P(DW) and the amount of tissue water (partial R2 = 0.74) and the log to the base 10 of triacylglycerol (partial R2 = 0.05). The P(WW) was associated with the log to the base 10 of triacylglycerol (partial R2 = 0.20), but no associations were found for P(NFWW). These findings indicate that decreased electrolyte content in dry and wet liver tissue with increased liver lipid content is predominantly due to the decrease in tissue water and therefore the distribution volume of electrolytes. In the longitudinal study, P(DW), P(WW), and P(NFWW) were decreased on d 14 p.p. Similar directional decreases were found for K, Mg, and Na, but P was the only electrolyte that was significantly decreased in liver tissue at d 14 p.p. This finding indicates that the P content of liver tissue decreases in early lactation due to a reduction in hepatocellular cytosol volume as well as a decrease in cytosolic P concentration, with the latter having biological relevance. The clinical significance of decreased cytosolic P concentration in the hepatocytes of dairy cows in early lactation remains to be determined.

Performance and metabolic and endocrine changes with emphasis on glucose metabolism in high-yielding dairy cows with high and low fat content in liver after calving.

Elevated liver fat content occurs in high-yielding dairy cows during the transition from pregnancy to lactation after fat mobilization and may affect hepatic glucose metabolism, but the degree of liver fat storage is highly variable. Therefore, we studied metabolic and endocrine changes and hepatic glucose metabolism in cows that markedly differ in liver fat content. Multiparous cows from the same herd with high (HFL; n = 10) and low (LFL; n = 10) liver fat contents (mean of d 1, 10, and 21 after calving for each cow, respectively) were studied from 60 d before expected calving to 56 d in milk. Cows were fed ad libitum and all cows received the same diets. Liver samples were taken on d 1, 10, and 21 after calving; mean fat content (+/-SEM) in liver of HFL cows was 174 +/- 9.6 mg/g, whereas mean liver fat content in LFL cows was 77 +/- 3.3 mg/g. Blood samples were taken 20 and 7 d before expected calving and 0, 7, 14, 28, and 56 d after calving to measure plasma concentrations of nonesterified fatty acids, beta-hydroxybutyrate, glucose, insulin, glucagon, insulin-like growth factor-I, and leptin. In liver, glycogen content as well as mRNA levels of phosphoenolpyruvate carboxykinase, pyruvate carboxylase, glucose-6-phosphatase, and glucose transporter were measured by quantitative real-time PCR. Back fat thickness decreased and dry matter intake increased with onset of lactation, and back fat thickness was higher but dry matter intake was lower in HFL than in LFL. Energy-corrected milk yield did not differ between groups, but milk fat content was higher and lactose content was lower in HFL than LFL at the beginning of lactation. Energy balance was more negative in HFL than in LFL. Plasma nonesterified fatty acids and beta-hydroxybutyrate concentrations increased and plasma glucose concentration tended to decrease more in HFL than LFL with onset of lactation. Glucagon to insulin ratios increased more in HFL than LFL with onset of lactation. Hepatic glycogen content was higher in LFL than HFL, whereas mRNA levels of glucose-6-phosphatase and pyruvate carboxylase were higher in HFL than in LFL, and cytosolic phosphoenolpyruvate carboxykinase mRNA level increased similarly after parturition in both groups. In conclusion, an elevated liver fat content was related to greater fat mobilization and reduced feed intake and was associated with effects on hepatic glucose metabolism. As environment and feeding management were the same, individual cow factors were responsible for differences in energy metabolism during the transition period.

Prepartal concentration of estradiol-17beta in heifers with stillborn calves.

This study was conducted to investigate hormonal imbalances preceding stillbirths and dystocia in primiparous heifers. The study was conducted between 2003 and 2004 on a German dairy farm, including 433 heifers. Starting 3 wk before calving, a weekly blood sample was collected. At calving, another blood sample was obtained, and the calving ease (grade 0 = unassisted to grade 2 = heavy pull with mechanical calf puller), sex, birth weight, as well as vitality status (stillborn, alive) of the calf were recorded. The blood serum was analyzed for estradiol-17beta and progesterone concentration. At parturition, the measured estradiol-17beta concentration was greater in heifers delivering bulls than in those with female calves and was increasing with greater birth weight of the calf and increasing calving difficulty score. Already 2 wk before calving, the serum estradiol-17beta concentration was significantly smaller in heifers with stillborn than live calves. On the other hand, the progesterone concentration was greater 2 wk before calving in heifers with stillborn calves, but it was unaffected by the birth weight or sex of the calf or the calving difficulty score. Stillborn and live calves did not differ in birth weight or pregnancy duration. The smaller estradiol-17beta concentrations of the heifers with stillborn calves could indicate an abnormality of the placenta or an abnormality of hormonal signals from the calf to the placenta in the weeks before the calving.