Roughage

The experiment started individually for each cow with the expected day 42 ante partum (ap) and ended at day 110 pp. A total of 59 pluriparous German Holstein dairy cows, including eight rumen- and duodenum-cannulated cows, were assigned to two groups, a control (CON, n = 30) and an L-carnitine group (CAR, n = 29), balanced for numbers of lactation (2–5 lactations), body weight (568–1008 kg), body condition score (2.5–4.75) and fat-corrected milk yield of previous lactation. To circumvent ruminal degradation, the cows in CAR received 125 g of a rumen-protected L-carnitine product (Carneon 20 Rumin-Pro, Kaesler Nutrition GmbH, Cuxhaven, Germany) per cow and day, which was included in the concentrate feed. This amount corresponded to a daily L-carnitine intake of 25 g per cow and day. To balance the fat content of the L-carnitine product, CON obtained an equivalent fat product (BergaFat F-100 HP, Berg+Schmidt GmbH & Co. KG, Hamburg, Germany) as used for the rumen protection of the L-carnitine. The cows were kept in a free-stall barn with slatted floors and cubicles with rubber pads and were rehoused for calving in the calving pen, where a maximum of two cows were kept in one straw bedding box.
Both groups were fed with a partial mixed ration (PMR). Whereas the composition of roughage remained unchanged during the whole trial (70% maize silage and 30% grass silage), the proportion of roughage to concentrate was variable in accordance with the recommendation of nutrient and energy supply of the Society of Nutrition Physiology (GfE). Initially, up to day 1 ap, diets of 80% roughage and 20% concentrate were fed. The amount of concentrate was increased from 30% to 50% up to 14 days pp and from then on, 50% concentrate was constantly fed up to day 110 pp. The PMR was offered by feed-weigh troughs (Roughage Intake Control, System Insentec B.V., Marknesse, The Netherlands) and the supplementary, restricted, pelleted concentrate was provided via concentrate feeding stations (Insentec B.V., Marknesse, The Netherlands). Water was offered for ad libitum intake. The components and the chemical composition of roughages and concentrate feed are shown in Table 1.

Sheep were trained over a period of two months to undertake a judgement bias test (see below), animals remained on pasture during the training period. The 30 sheep that reached the learning criteria in the judgement bias test (see below) were pseudo-randomly split into two groups of 15 animals each, which were allocated to chronic stress and control treatments.
The chronic stress group was housed in individual pens (0.9 x 2 m) in an indoor sheep facility and restricted from lying comfortably for 18 h/day for a period of nine days. An uncomfortable lying surface comprised of a 0.9 x 2 x 0.14 m wooden grid was placed on the floor in each pen during the lying restricted times. Previous research has shown that seven days of lying restriction was sufficient to induce chronic stress response in dairy cows [30 (link)]. A vacant pen between each sheep prevented physical interactions but permitted visual and auditory contact between animals. Sheep were moved into an adjacent pen without the grids for a six hour interval each day at unpredictable times (often during daylight hours), thus allowing access to a metal mesh floor to lie on. Contact between handlers and animals was kept to a minimum and restricted to the most necessary husbandry procedures (e.g., feeding, weighing, etc). The chronic stress group also received an additional 200 g of oaten chaff (total 400 g oaten chaff) in order to ensure sufficient roughage intake and to compensate for the possibility of a small amount of grass intake in the control group (see below).
The control group was kept in a paddock grazed bare of feed (in order to prevent confounding effects of diet) adjacent to the judgement bias test facility. Control sheep also received two small palatable rewards (total 50 g of a lupin and barley mixture) per day; once after an experimental manipulation had been applied (e.g., a blood sample had been collected) and once at a different time while in the paddock. The animals were handled gently (i.e., use of calm voice, no sudden movements, quiet environment etc) and were allowed to interact positively with the handlers if the sheep chose to do so (e.g., voluntary grooming).
The measurement protocol is summarised in Table 1. Commencement of chronic stress and control treatments was called day 1. Activity sensors (IceQubes, IceRobotics, South Queensferry, UK) were fitted to the right front leg of the Control group (day 2 and 3) and the chronic stress group (day 4 and 5) to measure the time spent lying. As insufficient devices were available to measure lying behaviour of all animals simultaneously, a serial design was used so that we could ensure that data was collected during the same daytime hours (between 10:00 h and15:00 h comfort time and between 16:00 h and 8:00 h the next day for restricted time) for both treatments (the chronic stress group was lying restricted during this period on all other days to induce the chronic stress, see Table 1) on 2 different days to account for variation in diurnal patterns of activity with time of day and also between days. Judgement bias was measured on day 6 and a combined CRH/AVP challenge was conducted on day 7. On day 8, attention bias was assessed and after this sheep were fitted with intravaginal temperature loggers. On day 9, sheep were exposed to shearing as an acute stressor. Body temperatures, ACTH and cortisol concentrations were monitored before and after shearing, and immediately after shearing judgement bias was assessed. Animals continued to receive their chronic stress or control treatments until all testing was completed at the end of day 9.

Forty heifers (16.7±1.3 months age, initial body weight [BW] 318.5±27.9 kg) were randomly allocated into three experimental groups according to the target daily weight gain, consisting of five animals per pen in a total of 8 pens (T-0.2, 2 pens; T-0.4, and −0.6, 3 pens) based on similar BW and age in months. The experiment was conducted for a total of 63 days. The appropriate average daily gain (ADG) for heifers suggested by National Institute of Animal Science (NIAS; 2012, 2017) [1 ,2 ] is 0.55 kg/d. Therefore, the target ADG in this experiment was set at 0.2, 0.4, and 0.6 kg/d, respectively. Feeds were offered according to the target ADG based on NIAS (2017) [2 ], and there was no adaptation period because the animals were fed the same mixed concentrates (formula feed) and roughage (rice straw) that were previously consumed. In order to minimize hunger stress of T-0.2 and −0.4, the feeding ratio of rice straw was set to 55%, 50%, and 45%, respectively, so that there was no difference in DM intake (DMI) among treatment groups, but the energy and protein levels in the feed became different. Feeds were offered twice equally at 07:00 and 17:00 h daily. Animals could access fresh water and mineral block without any restriction during the whole period. The nutrient content of the feed used in the experiment and the mixing ratio of the formulated feed are shown in Table 1. The BW was measured at the initial stage of the experiment, the second (36 d), and the end of the experiment (63 d). The feed intake was measured every week for calculating feed conversion ratio (FCR; feed/gain).

Roughage-rich diets have been shown to lessen the risk of obesity, CV disease, and several malignancies. DF shows the chances of reduction of several diseases that occurs due to lack of fiber in meat products after incorporating DF from different fruit and vegetable sourced wastes. Foods high in DF have a bulking effect. Over the past few years, an increasing body of research has focused on how to reap the health benefits of DF, mainly by adding fiber to a diet that is otherwise deficient in recent diet due to highly processed food product. DF acts as a laxative, reduces blood cholesterol, and provides antioxidant activity; these are just three of the many benefits that have been tallied during the last half-century. Some DF can’t do what they are credited with doing, while others can’t do it as well or as often.

Animals in all groups were fed by group feeding, and the tested diets were offered in equal amounts for all groups twice daily at 8 a.m. and 3 p.m., while fresh water was always available. The offered amounts of CFM and roughage were adjusted every 2 weeks according to changes in the animals' body weight. Diets were formulated to meet the requirements of growing goats according to (21 ).