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Background The aim of this study was to investigate if calves’ play behaviour and non-nutritive sucking behaviour, as indirect measures of welfare status, are associated with the age of the calf when group housed, age when observed, age difference within the group, pen size, milk feeding system, current or previous sicknesses, access to dry teat, indoor/outdoor rearing, sex, organic/conventional farm, group size and regrouping events. An observational study was conducted on 176 Danish dairy calves in the age range of 1–12 weeks, on both conventional (n = 17) and organic (n = 5) farms. All calves had been group housed before 8 weeks of age and had spent various periods of time with the dam and/or individually housed before being group housed. Behaviour was recorded continuously by filming each individual calf over a period of 30 min. Results The calf’s age when group housed for the first time was not found to be significantly associated with duration of either play behaviour (P = 0.55) or non-nutritive sucking behaviour (P = 0.44). It was found that calves had significantly reduced odds of playing for longer than the mean play duration (5.5 s) for each day of their lives (OR = 0.97, P = 0.003). Also, they had reduced odds of performing non-nutritive sucking behaviour for longer than the mean non-nutritive sucking duration (145.5 s) when milk was allocated by drinker buckets fitted with a teat compared to by bowl or trough (OR = 0.06, P = 0.02). Conclusion No significant associations were found between calves’ age when group housed for the first time and play and non-nutritive sucking behaviour. It was found that calves’ play behaviour decreased with increasing age, and that non-nutritive sucking behaviour decreased when milk was allocated with a teat compared to no teat.

The extraction of bioactive compounds and cellulose saccharification are potential directions for the valorization of spent mushroom substrate (SMS). Therefore, investigating the suitability of different extraction methods for recovering bioactive compounds from SMS and how the extraction affects the enzymatic saccharification is of uppermost relevance. In this work, bioactive compounds were extracted from Pleurotus spp. SMS using four extraction methods. For Soxhlet extraction (SoE), a 40:60 ethanol/water mixture gave the highest extraction efficiency (EE) (69.9–71.1%) among the seven solvent systems assayed. Reflux extraction with 40:60 ethanol/water increased the extraction yield and EE compared to SoE. A shorter reflux time yielded a higher extraction of carbohydrates than SoE, while a longer time was more effective for extracting phenolics. The extracts from 240 min of reflux had comparable antioxidant activity (0.3–0.5 mM GAE) with that achieved for SoE. Ultrasound-assisted extraction (UAE) at 65 °C for 60 min allowed an EE (~82%) higher than that achieved by either reflux for up to 150 min or SoE. Subcritical water extraction (SWE) at 150 °C resulted in the best extraction parameters among all the tested methods. Vanillic acid and chlorogenic acid were the primary phenolic acids identified in the extracts. A good correlation between the concentration of caffeic acid and the antioxidant activity of the extracts was found. Saccharification tests revealed an enhancement of the enzymatic digestibility of SMS cellulose after the extraction of bioactive compounds. The findings of this initial study provide indications on new research directions for maximizing the recovery of bioactive compounds and fermentable sugars from SMS.

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