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Hair cortisol concentration (HCC) might represent a promising marker for retrospective welfare assessment of dairy cows. The objective of the study was to explore the dynamics of HCC in diseased and healthy cows from eight-week ante partum (AP) to eight-week post partum (PP). Twenty-four pregnant cows were followed from drying off to week eight PP. Tail hair was used to measure cortisol at five different time points. The occurrence of peripartum diseases, lameness and the body condition score (BCS) were monitored on a weekly basis. Blood β-hydroxybutyric acid, non-esterified fatty acids, calcium and insulin-like growth factor-1 (IGF-1) concentrations were measured. The temperature-humidity index (THI) was continuously recorded. The median values of HCC in all cows were 0.4, 0.3, 0.6, 0.8 and 0.5 pg/mg at weeks eight, four AP, calving, weeks four, eight PP, respectively. There was no association between HCC and the occurrence of peripartum diseases (P ≥ 0.05). A positive correlation between HCC and BCS loss (P

Newborn ungulates depend on the timely supply of colostrum containing sufficient immunoglobulins to obtain passive immunity against disease. Brix refractometry enables a rapid on-farm estimation of colostrum quality and has been intensively studied in bovines. However, the suitability of Brix refractometers for assessing colostrum quality in goats and ewes has been scarcely evaluated. The present study compared bovine, caprine, and ovine colostrum quality estimation using an optical Brix refractometer. In addition, between-species variations in the relationships between Brix values and colostrum constituents (IgG, fat, protein, and lactose) and the accuracy of Brix refractometry at different cutoff values were evaluated by a receiver operating characteristic curve analysis. We measured the Brix value and contents of IgG, fat, protein, and lactose in 324 colostrum samples (108 cows, 116 does, and 100 ewes). Thresholds for classification of good colostrum quality (as determined by ELISA) were set at 50 mg IgG/mL in cows and 20 mg/mL in does and ewes. Bovine colostrum showed the greatest IgG concentrations compared with caprine and ovine colostrum. Fat and protein content was higher in sheep colostrum compared with the other species, whereas the highest lactose concentrations were detected in goat colostrum. Brix values ranged from 11.4 to 34.6% (22.1 ± 4.2%; mean ± standard deviation), 15.4 to 40.0% (28.5 ± 6.8%), and 8.8 to 39.8% (21.6 ± 5.3%) in bovine, ovine, and caprine colostrum, respectively. In all 3 species, Brix was highly correlated with IgG and protein concentrations (cows, r = 0.83 and 0.98; goats, r = 0.83 and 0.89; sheep, r = 0.75 and 0.87). Optimal cutoff points for greatest accuracy of Brix measurements were 19.3% Brix in cows [with 87.1% sensitivity (Se) and 100% specificity (Sp)], 20.7% Brix in does (with 53.5% Se and 100% Sp), and 26.5% Brix in ewes (with 75% Se and 91.3% Sp). In conclusion, Brix refractometry is an acceptable tool for on-farm estimations of colostrum quality in does and ewes despite distinct between-species variations in colostrum composition.

Glucocorticoids, particularly cortisol and corticosterone, are key homeostatic regulators during metabolic and endocrine adaptations including inflammatory responses. Besides the established response of total cortisol (TC) concentrations during inflammatory processes in dairy cows, we investigated changes of corticosterone, free cortisol (FC), and serum albumin as the main protein of unspecific cortisol binding, in response to an intramammary lipopolysaccharide (LPS) challenge. Furthermore, we evaluated relationships of glucocorticoid responses with concomitant alterations of metabolites and their endocrine regulators, insulin and glucagon. Blood samples of 10 multiparous Holstein dairy cows (26.8 ± 3.4 d in milk, previous lactation yield: 7,601 ± 938 kg; mean ± SD) were obtained every 30 min up to 5 h after the LPS instillation, and rectal temperature and heart rate were measured in parallel. Corticosterone was measured by enzyme immunoassay, TC by radioimmunoassay, and the proportion of FC by ultrafiltration. A mixed model was used to evaluate differences within the investigated parameters among selected time points (0, 3.5, and 5 h relative to the intramammary LPS administration). Rectal temperature increased up to 41.6 ± 0.1°C at 5 h after the LPS application. Concentrations of TC and corticosterone increased until 3.5 h, and the proportion of FC relative to TC more than doubled until 3.5 h after LPS administration. Serum albumin concentration was reduced at 5 h compared with initial values, whereas concentrations of insulin, glucagon, and glucose were increased after 5 h compared with 0 h. In conclusion, the stimulation of the immune system by the intramammary LPS administration is accompanied by distinct metabolic and endocrine changes. Corticosterone and TC concentrations react similarly in response to the LPS challenge and earlier compared with metabolic adaptations. The increased need of active cortisol is covered by both increased secretion and a higher percentage of FC.

The intact blood-milk barrier (BMB) prevents an uncontrolled exchange of soluble and cellular components between blood and milk in the mammary gland. It enables the sustainability of the optimal milk composition for the nourishment of the offspring. Endothelial cells, connective tissue, the basal membrane, and mainly the epithelial cells provide the semipermeability of this barrier, allowing only a selective transfer of components necessary for milk production. The epithelial cells are closely connected to each other by different formations, in which the tight junctions are the most critical for separating the milk-containing compartments from the surrounding extracellular fluid and vasculature. During mastitis, the integrity of the BMB is reduced. This facilitates the transfer of immune cells and immune factors such as antibodies from blood into milk. Simultaneously, the transfer of soluble blood constituents without an obvious immune function into milk is promoted. Furthermore, a reduced BMB integrity causes a loss of milk constituents into the blood circulation. Different mechanisms are responsible for the barrier impairment including tight junction opening, but also cell degradation. To promote the cure of mastitis, the targeted manipulation of the BMB permeability may be a tool to optimize the immune function of the mammary gland. An intensified opening of the BMB supports the antibody transfer from blood into milk, which is supposed to increase the contribution of the specific immune system in the immune defense. On the contrary, a fast closure of the BMB during the recovery from mastitis can accelerate the normalization of milk composition and milk yield. Various agents have been experimentally shown to either open (e.g., pathogens and pathogen-associated molecular patterns, several nonsteroidal anti-inflammatory drugs, oxytocin, calcium chelators) or close (e.g., glucocorticoids, nonsteroidal anti-inflammatory drugs, natural anti-inflammatory drugs) the BMB.

The aim of the present study was to measure salivary cortisol concentrations of horses before and after hypothalamic-pituitary-adrenal (HPA) axis stimulation by means of liquid chromatography-tandem-mass spectrometry (LC-MS/MS) and an immunoassay (cELISA) for method comparison. Nine clinically healthy horses participated in the study. An ACTH stimulation test was performed. Saliva samples were collected before (T0) and 60 (T60) min after intravenous injection of 1 μg/kg BW synthetic ACTH1-24. LC-MS/MS was assessed for the determination of equine salivary cortisol. The results of these measurements were then compared to the results obtained by a cELISA, which has previously been validated for use in horses. The Pearson correlation coefficient was calculated and showed no correlation at T0 (r = -0.2452; P = 0.5249) and significantly correlated results at T60 (r = 0.8334; P = 0.0053). Bland-Altman-Plots of T60 revealed that immunoassay measurements led to higher outcome values than LC-MS/MS. On average, immunoassay results were 2.3 times higher. Poor agreement between both methods at T0 is potentially a consequence of inaccuracy in the very low measuring range of the immunoassay, and to a smaller extent, structurally similar cross-reacting agents and matrix effects, which might bias the results. Overestimation of immunoassay results at T60 might be due to different standardization of both methods, non-avoidable matrix effects on the antigen-antibody interaction in the ELISA, and possibly cross-reactions of other steroids. While immunoassay measurements of equine salivary cortisol yielded higher but reasonably correlated results for elevated cortisol concentrations after stimulation of the HPA axis, LC-MS/MS provided more accurate results, particularly for baseline cortisol concentrations close to the limit of detection of the ELISA.

Mastitis is responsible for substantial economic loss and significant animal welfare concerns for the dairy industry. Sensors that measure electrical conductivity (EC) and enzyme concentrations of lactate dehydrogenase (LDH) are presently used for automatic detection of mastitis. However, EC is not sensitive enough to detect mastitis, and the ability of LDH activity to identify mastitis caused by different pathogens is a potential option that needs to be investigated. This study was conducted to test the following hypotheses: (a) strict foremilk before milk ejection is more informative in detecting mastitis, in general, than foremilk removed after cows were stimulated for milk ejection; and (b) the value of LDH activity as a mastitis indicator depends on the type of pathogen associated with the infection. Milk samples (before afternoon milking) from 48 Holstein-Friesian cows at the University of Sydney's dairy farm (Camden, New South Wales, Australia) with EC > 7.5 mS/cm in any of the 4 quarters were collected over a period of 2 mo. Quarter milk samples (n = 343) from 48 cows were collected manually in the automatic milking rotary in 3 steps: foremilk before (strict foremilk) and after milk ejection, followed by an aseptic sample for bacteriological culture. The EC (mS), LDH (U/L), SCC (cells/mL), and milk protein and fat content (%) of foremilk in both sampling times were compared and used as predictors for gram-positive and gram-negative mastitis. Quarter (n = 515) observations from 44 cows were analyzed using a logistic mixed or linear mixed model, with cow and quarter nested within cow as random effects. Milk from both sampling times was also assessed by producing a receiver operating characteristic (ROC) curve and calculating the area under the curve (AUC) to determine ability to detect mastitis. Overall, EC and LDH were greater and milk protein (%) was lower in strict foremilk than in milk fractions obtained after milk ejection. Data from strict foremilk samples had slightly higher AUC values (0.98 to 0.99 vs. 0.97 to 0.98, respectively) than did the after-ejection milk samples. Although gram-negative coliform mastitis had significantly higher LDH activity than did gram-positive mastitis (6.19 vs. 5.34 log10 U/L), the robustness of this result is questionable due to limited sample size. We concluded that milk samples taken before ejection can influence major mastitis indicators, suggesting that automatic milking system sensors could be modified to monitor milk before ejection for more efficient mastitis detection.

Colostrum represents the sole source to acquire humoral immunity and is an important energy source for newborn lambs and goat kids. However, colostrum composition (i.e., the contents of IgG, fat, protein, and lactose) is affected by various factors such as parity and litter size and, potentially, by breed. In the present study, we examined the colostrum composition of different goat and sheep breeds raised for milk and meat production in Switzerland and Germany. Ten goat breeds (Anglo-Nubian, Appenzell, Boer, Bunte Deutsche Edelziege, Chamois-colored, Grisons Striped, Peacock, Saanen, Toggenburg, and Valais Blackneck) and 10 sheep breeds (Brown-Headed Meat, East Friesian Milk, German Blackheaded Mutton, Gray Horned Heath, Lacaune Dairy, Merino Land, Swiss Black-Brown Mountain, Swiss Charollais, Swiss White Alpine, and Valais Blacknose) were involved in this study. First colostrum samples were obtained from ewes (n = 100) and goats (n = 116) between 10 and 390 min after parturition and analyzed for total IgG, fat, protein, and lactose contents. Colostral IgG concentrations varied between 4.8 and 75.0 mg/mL in goats, and between 6.2 and 65.4 mg/mL in ewes, and the time interval between milking and parturition did not affect colostral IgG concentrations. In goats, the highest IgG concentrations were found in Boer (meat-type; 61.0 ± 10.3 mg/mL; mean ± SD) and the lowest concentrations were observed in Bunte Deutsche Edelziege (milk-type; 26.5 ± 12.5 mg/mL). In sheep, East Friesian Milk and Lacaune Dairy showed the lowest colostral IgG concentrations (17.9 ± 7.3 and 20.2 ± 8.0 mg/mL, respectively), and the highest values were observed in the Merino Land breed (44.2 ± 15.7 mg/mL). The lowest fat and protein concentrations and concomitantly highest lactose concentrations were observed in colostrum of East Friesian Milk and Lacaune Dairy sheep. Parity number did not affect colostrum composition in sheep or goats. In contrast, colostral fat content was higher in ewes bearing twins and triplets than in those carrying singletons. Increasing litter size tended to be associated with higher protein and lower lactose concentrations in ovine (i.e., singletons vs. twins vs. triplets) and caprine colostrum (i.e., singletons vs. twins), whereas colostral IgG concentrations were not affected by litter size. In conclusion, IgG and concentrations of other colostrum constituents showed a wide range in goats and ewes and were mainly affected by the type of breed.

Serotonin (5-hydroxytryptamine, 5-HT) affects many physiological functions because it is involved in glucose and lipid metabolism, calcium homeostasis, and regulation of lactation in dairy cows. This study aimed to examine physiological differences in serum 5-HT concentrations (high vs. low) and their association with metabolic status and milk production at the onset of lactation. Twelve multiparous Holstein dairy cows were milked within 4 h of calving, and blood and milk samples were collected at the first 6 subsequent milkings after parturition and at the evening milkings on d 5, 8, 10, and 14. Cows were retrospectively divided into 2 groups (6 cows/group): low serum 5-HT (LSS) and high serum 5-HT (HSS) according to their calculated areas under the curve (AUC) for serum 5-HT for the entire experimental period (cut-off: 46,000 ng/mL × 324 h). Concentrations of 5-HT, free fatty acids (FFA), β-hydroxybutyrate (BHB), glucose, calcium, and IGF-1 were measured in blood. Milk was analyzed for fat, protein, lactose, and 5-HT concentrations. Milk yield was recorded at each milking and energy-corrected milk yield was calculated. Serum 5-HT concentrations were higher in HSS than in LSS [AUC (ng/mL × 324 h): 57,830 ± 4,810 vs. 25,005 ± 5,930]. The amount of energy-corrected milk was lower in HSS than in LSS. The HSS group produced less colostrum and had decreased milk yield, specifically during the first 6 milkings. Concentrations of FFA, BHB, and glucose in plasma did not differ between groups. Concentrations of IGF-1 in serum were elevated in HSS compared with LSS throughout the experiment. Total circulating calcium concentrations in serum tended to be higher in HSS than in LSS. Milk fat and protein yields were decreased in HSS compared with LSS. Milk 5-HT decreased overall during the experimental period, with LSS maintaining higher 5-HT concentrations than HSS until d 14 of lactation. In conclusion, cows with high serum 5-HT concentrations showed a reduced metabolic load at the onset of lactation, concomitantly lower milk yield, and a reduced energy output via milk.