High-resolution daily profiles of tissue adrenal steroids by portable automated collection.

Rhythms are intrinsic to endocrine systems, and disruption of these hormone oscillations occurs at very early stages of the disease. Because adrenal hormones are secreted with both circadian and ultradian periods, conventional single-time point measurements provide limited information about rhythmicity and, crucially, do not provide information during sleep, when many hormones fluctuate from nadir to peak concentrations. If blood sampling is attempted overnight, then this necessitates admission to a clinical research unit, can be stressful, and disturbs sleep. To overcome this problem and to measure free hormones within their target tissues, we used microdialysis, an ambulatory fraction collector, and liquid chromatography-tandem mass spectrometry to obtain high-resolution profiles of tissue adrenal steroids over 24 hours in 214 healthy volunteers. For validation, we compared tissue against plasma measurements in a further seven healthy volunteers. Sample collection from subcutaneous tissue was safe, well tolerated, and allowed most normal activities to continue. In addition to cortisol, we identified daily and ultradian variation in free cortisone, corticosterone, 18-hydroxycortisol, aldosterone, tetrahydrocortisol and allo-tetrahydrocortisol, and the presence of dehydroepiandrosterone sulfate. We used mathematical and computational methods to quantify the interindividual variability of hormones at different times of the day and develop "dynamic markers" of normality in healthy individuals stratified by sex, age, and body mass index. Our results provide insight into the dynamics of adrenal steroids in tissue in real-world settings and may serve as a normative reference for biomarkers of endocrine disorders (ULTRADIAN, NCT02934399).

Assessing dairy cow welfare during the grazing and housing periods on spring-calving, pasture-based dairy farms.

The different periods characterizing spring-calving, pasture-based dairy systems common in Ireland have seldom been the focus of large-scale dairy cow welfare research. Thus, the aim of this study was to devise and conduct an animal-based welfare assessment during both the grazing and housing periods on spring-calving, pasture-based dairy farms, to identify areas for improvement and establish benchmarks for indicators of good welfare. Assessment of seven animal-based welfare indicators was conducted during two visits (one each at grazing and housing) to 82 commercial dairy farms in southern Ireland. Herd-level descriptive statistics were performed for all welfare indicators at each visit, and differences between visits were analyzed using paired t-tests and Wilcoxon signed-rank tests. A mean of 9% and 10% clinically lame cows (mobility scores 2 and 3) were observed at housing and grazing, respectively. Recommended body condition scores (BCS) were not met for a mean of 13% of cows at grazing and 23% at housing, with more over-conditioned cows present at housing than grazing (P < 0.001). Ocular discharge was uncommon in both periods. Prevalence of moderate and severe nasal discharge combined was lower during housing (5%) than grazing (7%). In both periods, similar mean levels of tail injury were observed: 2% to 3% of cows with tail lacerations, 9% with broken tails, and 8% (measured at housing only) with docked tails. Integument alterations involved primarily hair loss and were most prevalent on the hindquarters (26%) during grazing and on the head-neck-back (66%) and the hindquarter (32%) regions during housing. Cows displayed an avoidance distance of >1 m (indicative of a fearful response) from an approaching human in an average of 82% of grazing cows and 42% to 75% of housed cows, dependent on test location. Opportunities to improve welfare in this system were identified in the areas of tail injury prevention, nasal health, and the management of indoor housing and feeding. The performance of the top 20% of farms for each welfare indicator was used to establish benchmarks of: 0% to 5% clinical lameness, 0% to 12% of cows outside recommended BCS, 0% to 27% ocular discharge, 2% to 16% nasal discharge, 0% tail lacerations and docked tails, 0% to 3% tail breaks, 0% to 14% integument alterations, and 4% to 74% for avoidance distance of >1 m. These represent attainable targets for spring-calving pasture-based farms to promote good dairy cow welfare.

Synchronization of Dairy Cows Does Not Limit the Behavioral Response to Treatment in Mixed Treatment Experimental Designs.

In many research studies, animals assigned to different treatments are housed adjacently or together in a group. One critique of these designs has been the potential role of behavioral facilitation and synchronization between animals on different treatments in mixed treatment groups (heterogeneous groups). To evaluate this, we compared the synchrony of feeding behavior between dairy cows housed in heterogeneous groups to cows exposed to the same treatment simultaneously (homogenous groups). Twenty-four cows were exposed to each of the two treatments over 21 days in a replicated cross-over design. Treatments were two different schedules of timing of feed delivery: (A) feeding at milking time and (B) feeding halfway between milking times. For the last 7 days of each treatment period, feeding behavior was recorded electronically. Kappa coefficients were calculated for each animal within each group, as an estimate of agreement that any two cows within a group (i.e., each individual and each other cow in her group) would both be engaged in feeding activity for any hour of the day. The level of synchrony was similar for cows within homogenous groups (kappa = 0.31 ± 0.030) compared with cows on the same treatment within heterogeneous groups (kappa = 0.32 ± 0.037). Within heterogeneous groups, cows on the same treatment were nearly 50% more synchronized with each other than with those on the other treatment (kappa = 0.22 ± 0.029). These results suggest that synchronization of feeding behavior does not restrict our ability to impose different treatments on individual cows within a group.