Bioactive metabolites of Asparagopsis stabilized in canola oil completely suppress methane emissions in beef cattle fed a feedlot diet.

Asparagopsis taxiformis (Asparagopsis) has been shown to be highly efficacious at inhibiting the production of methane (CH4) in ruminants. To date, Asparagopsis has been primarily produced as a dietary supplement by freeze-drying to retain the volatile bioactive compound bromoform (CHBr3) in the product. Steeping of Asparagopsis bioactive compounds into a vegetable oil carrier (Asp-Oil) is an alternative method of stabilizing Asparagopsis as a ruminant feed additive. A dose-response experimental design used 3 Asp-Oil-canola oil blends, low, medium, and high Asp-Oil which provided 17, 34, and 51 mg Asparagopsis derived CHBr3/kg dry matter intake (DMI), respectively (in addition to a zero CHBr3 canola oil control), in a tempered-barley based feedlot finisher diet, fed for 59 d to 20 Angus heifers (five replicates per treatment). On four occasions, live weight was measured and CH4 emissions were quantified in respiration chambers, and blood, rumen fluid, and fecal samples were collected. At the end of the experiment, all animals were slaughtered, with carcasses graded, and samples of meat and edible offal collected for testing of consumer sensory qualities and residues of CHBr3, bromide, and iodide. All Asp-Oil treatments reduced CH4 yield (g CH4/kg DMI, P = 0.008) from control levels, with the low, medium, and high Asp-Oil achieving 64%, 98%, and 99% reduction, respectively. Dissolved hydrogen increased linearly with increasing Asp-Oil inclusion, by more than 17-fold in the high Asp-Oil group (P = 0.017). There was no effect of Asp-Oil treatment on rumen temperature, pH, reduction potential, volatile fatty acid and ammonia production, rumen pathology, and histopathology (P > 0.10). There were no differences in animal production and carcass parameters (P > 0.10). There was no detectable CHBr3 in feces or any carcass samples (P > 0.10), and iodide and bromide residues in kidneys were at levels unlikely to lead to consumers exceeding recommended maximum intakes. Overall, Asp-Oil was found to be safe for animals and consumers of meat, and effective at reducing CH4 emissions and yield by up to 99% within the range of inclusion levels tested.

Red seaweed, Asparagopsis taxiformis (Asparagopsis), has been shown to be highly effective at inhibiting the production of methane (CH4) in ruminants. An alternative to feeding whole, freeze-dried Asparagopsis is steeping the biomass in vegetable oil to stabilize the bioactive compounds (Asp-Oil) and feeding Asp-Oil to ruminants as a component of their dietary intake. This experiment measured the CH4 reduction potential and safety of Asp-Oil in a trial with 20 Angus heifers, fed iso-fat feedlot diets containing one of the three levels of Asp-Oil, or a control oil. Compared to the control, bromoform inclusion levels of 17, 34, and 51 mg/kg of dry matter (DM; low, medium, high) reduced CH4 yield (g CH4/kg DM intake) by 64%, 98%, and 99%, respectively. There were no effects on animal production or carcass characteristics. There were no impacts on animal health, welfare, or rumen function. Carcasses were safe for human consumption, and there was no bromoform detected in any carcass samples. Overall, Asp-Oil was found to effectively reduce CH4 emissions and is safe for animals and consumers of meat and edible offal.

The effects of stocking density on behavior and biological functioning of penned sheep under continuous heat load conditions.

Stocking density may impact sheep welfare during live export voyages that occur under hot and humid conditions. The aim of this study was to assess the welfare implications for sheep housed at three allometric stocking densities (k = 0.030, 0.033, 0.042), while exposed to hot and humid climatic conditions. For 21 d, Merino wethers (n = 216) were housed in 12 pens of 18 wethers, in two climate-controlled rooms where wet-bulb temperature (TWB) mimicked the conditions of a live export voyage with high heat and humidity, and limited diurnal variation. Scan sampling of standing and lying behaviors was conducted on days 2, 5, 8, 11, 15, 18, and 20, at hourly intervals. Agonistic interactions were scored continuously on the same days between 1750 and 1800 h. Liveweights were recorded at the start and end of the study. For a subset of focal wethers (3 per pen), whole blood variables were assessed at the start and end of the experiment, along with fecal glucocorticoid metabolites (FGCM), which were also assessed on days 7 and 14. Rumen temperatures (TRUM) of focal wethers were recorded at 10-min intervals, and their respiration rates (RR) were measured every 2 h on days 1, 3, and from days 7 to 21. Focal wethers were slaughtered for necropsy after the study, and both adrenal glands were excised and weighed. The expression of some lying positions was impaired at high stocking densities, and lying with outstretched legs increased at high TWB. For respiration rates, there was an interaction between stocking density and TWB, such that RR was reduced by the provision of additional space at high TWB. TRUM was relatively unaffected by stocking density but increased at higher TWB, and any effects of stocking density on FGCM concentrations, liveweights (LW), adrenal gland weights or blood variables were minimal. Necropsy examination showed no indication that the wethers had experienced ongoing respiratory distress. These results suggest that the wethers were able to cope with these increases in stocking density under the conditions imposed. However, based on this evidence, the provision of additional space under hot conditions may be beneficial to facilitating the expression of some lying positions. Whilst the experiment was designed to emulate certain conditions relevant during live export voyages, other factors that may induce stress during this mode of transport were not present, and so the conclusions must be interpreted in the context of the experimental conditions.

There is a high demand for Australian sheep to be exported to the Middle East, and for live export voyages which depart during an Australian winter, heat, and humidity increase rapidly as ships cross the equator and approach destination countries. Concern about sheep becoming heat stressed during these voyages has increased, and industry attention has focused on the potential role of stocking density in determining heat stress risk in this context. High stocking densities limit the body surface area available for heat loss and can increase heat exchange between individual sheep. This study aimed to assess the welfare implications of three stocking densities, for sheep exposed to climatic conditions similar to those experienced during a live export voyage to the Middle East. Higher stocking densities restricted the ability of sheep to lie in some positions, but stocking density had limited effects on heat stress indicators or the physiology of the sheep. These results suggested that the sheep were able to cope with these increases in stocking density under the conditions imposed, but the conclusions must be interpreted in the context of the controlled experimental conditions.

Effect of 3-nitrooxypropanol on enteric methane emissions of feedlot cattle fed with a tempered barley-based diet with canola oil.

A dose-response experiment was designed to examine the effect of 3-nitrooxypropanol (3-NOP) on methane (CH4) emissions, rumen function and performance of feedlot cattle fed a tempered barley-based diet with canola oil. Twenty Angus steers of initial body weight (BW) of 356 ±â€…14.4 kg were allocated in a randomized complete block design. Initial BW was used as the blocking criterion. Cattle were housed in individual indoor pens for 112 d, including the first 21 d of adaptation followed by a 90-d finishing period when five different 3-NOP inclusion rates were compared: 0 mg/kg dry matter (DM; control), 50 mg/kg DM, 75 mg/kg DM, 100 mg/kg DM, and 125 mg/kg DM. Daily CH4 production was measured on day 7 (last day of starter diet), day 14 (last day of the first intermediate diet), and day 21 (last day of the second intermediate diet) of the adaptation period and on days 28, 49, 70, 91, and 112 of the finisher period using open circuit respiration chambers. Rumen digesta samples were collected from each steer on the day prior to chamber measurement postfeeding, and prefeeding on the day after the chamber measurement, for determination of rumen volatile fatty acids (VFA), ammonium-N, protozoa enumeration, pH, and reduction potential. Dry matter intake (DMI) was recorded daily and BW weekly. Data were analyzed in a mixed model including period, 3-NOP dose and their interaction as fixed effects, and block as a random effect. Our results demonstrated both a linear and quadratic (decreasing rate of change) effect on CH4 production (g/d) and CH4 yield (g/kg DMI) as 3-NOP dose increased (P < 0.01). The achieved mitigation for CH4 yield in our study ranged from approximately 65.5% up to 87.6% relative to control steers fed a finishing feedlot diet. Our results revealed that 3-NOP dose did not alter rumen fermentation parameters such as ammonium-N, VFA concentration nor VFA molar proportions. Although this experimental design was not focused on the effect of 3-NOP dose on feedlot performance, no negative effects of any 3-NOP dose were detected on animal production parameters. Ultimately, the knowledge on the CH4 suppression pattern of 3-NOP may facilitate sustainable pathways for the feedlot industry to lower its carbon footprint.

Livestock methane (CH4) is the main source of greenhouse gases (GHGs) in agriculture, contributing to 11.6% of global GHGs emissions from human-related activities. Therefore, mitigating CH4 emissions from ruminant animals is a great opportunity for meeting the current climate targets. In this experiment, increasing inclusion rates of a promising CH4-mitigating compound, 3-nitrooxypropanol (3-NOP, from 50 to 125 mg of 3-NOP/kg of dry matter [DM]), were added to a barley-based feedlot diet containing 25 ppm of monensin and 7% fat (DM-basis) and fed to Angus steers. Under these conditions, increasing inclusion rate of 3-NOP reduced both production and yield of CH4 by up to 90%. Rumen fermentation, feed intake, and average daily gain were not affected by the 3-NOP dose. Our results on the potential CH4 suppression of 3-NOP may assist the feedlot industry towards sustainability by lowering its GHG output.

The effects of antibiotic-free supplementation on the ruminal pH variability and methane emissions of beef cattle under the challenge of subacute ruminal acidosis (SARA).

Subacute ruminal acidosis (SARA) in feedlot cattle during the feed transition to grain-based diets is a significant constraint to animal health and productivity. This experiment assessed an antibiotic-free supplement (ProTect®) effects on ruminal pH variability and methane (CH4) emissions of cattle during the challenge of SARA. Ten 18-month-old Angus steers (472 ± 4.8 kg) were randomly allocated into monensin (n = 5) and ProTect® groups (n = 5) and progressively introduced to grain diets incorporating monensin or ProTect® for 36 days of the experiment [starter (7 days; 45% grain), T1 (7 days; 56% grain), T2 (7 days; 67% grain), finisher (15 days; 78% grain)]. The pH variability on the finisher period was reduced by the ProTect® supplement (6.6% vs. 5.2%; P < 0.01), with CH4 emissions being significantly higher relative to the monensin group [88.2 g/day (9.3 g CH4/kg DMI) vs. 133.7 g/day (14.1 g CH4/kg DMI); P < 0.01]. There was no difference between treatments in the time spent on the ruminal pH < 5.6 or < 5.8 (P > 0.05). The model evaluation for the ruminal pH variation indicated that the mean absolute error (MAE) proportion for both groups was good within the same range [4.05% (monensin) vs. 4.25% (ProTect®)] with identical root mean square prediction error (RMSPE) (0.34). It is concluded that the ProTect® supplement is an effective alternative to monensin for preventing SARA in feedlot cattle by managing ruminal pH variation during the transition to high-grain diets. Both monensin and ProTect® supplemented cattle exhibited lower CH4 yield compared to cattle fed forages and low-concentrate diets.

Corrigendum: Stocking density, restricted trough space, and implications for sheep behaviour and biological functioning.

[This corrects the article DOI: 10.3389/fvets.2022.965635.].

Stocking density, restricted trough space, and implications for sheep behaviour and biological functioning.

Stocking density and trough space allowance can potentially impact sheep welfare during live export voyages. The aim of this study was to assess the welfare implications for sheep housed at five allometric stocking densities, with either unrestricted or restricted trough space allowance. Merino wethers (n = 720) were housed in 40 pens of 18 heads for 18 days. Two 5-min continuous focal animal observations (n = 3/pen) were conducted on days 3, 5, 11, and 17. Scan sampling of standing and lying behaviours were conducted on the same days at hourly intervals. Live weights and immune cell counts were quantified at the start and end of the experiment, as well as faecal glucocorticoid metabolites (FGCMs), which were also assessed on days 6 and 12. Focal animals housed at higher stocking densities spent less time lying during one of the continuous observation periods, but no important effects on the overall number of animals lying or on the synchronicity of lying were evident. The scan sampling results indicated that the expression of some preferred lying positions was impaired at high stocking densities, and that high stocking densities also resulted in increased agonistic social interactions and displacement events at the start of the trial. There was a slight reduction in day 18 live weights for animals housed at higher stocking densities, but FGCM concentrations and immune cell counts were essentially unaffected. Trough space had no important effects on day 18 live weight, FGCM concentrations, or immune cell counts, and had limited effects on sheep behaviour. The lack of important impacts on biological fitness traits suggests that the behavioural responses observed were sufficient in allowing sheep to cope with their environment. However, we provide evidence that the provision of additional space is beneficial in reducing the time it takes for animals to adapt to their environment and to facilitate the expression of some preferred lying positions. While designed to emulate certain conditions relevant during live export voyages, some factors that may induce stress during this mode of transport were not present such as heat and ocean swell, so the conclusions must be interpreted in the context of the experimental conditions.

Feedlot Factors Influencing the Incidence of Dark Cutting in Australian Grain-Fed Beef.

It has been well-established that dark cutting (DC) is a multifactorial issue that is associated with numerous animal and management factors. However, there is limited understanding of the feedlot-based factors that contribute to the influence of DC. The aim of this study was to evaluate the effect of climate, animal, and feedlot factors on the incidence of pH non-compliance in Australian grain-fed cattle. For this study, feedlot and abattoir records from 142,228 individual cattle over a 1-year period were investigated. These data incorporated records from seven feedlots that consigned cattle to three abattoirs. The average incidence of DC in these carcasses was 2.8%. The production factors that were associated with increased risk of DC included feedlot, sex, hormone growth promotants (HGP), cattle health, and days on feed (DOF). Additionally, DC also increased by reduced solar radiation (SR, W/m2), lower wind speeds (WS, m/s), increased ambient temperature (TA, °C), higher rainfall, a higher average temperature-humidity index (THI), and increased duration of time above heat-load-index threshold of 86 (HLI ≥ 86) during the 7 days prior to feedlot departure. This study identified the feedlot factors that increase the risk of DC from a feedlot-management perspective.

A Pilot Study Using Accelerometers to Characterise the Licking Behaviour of Penned Cattle at a Mineral Block Supplement.

Identifying the licking behaviour in beef cattle may provide a means to measure time spent licking for estimating individual block supplement intake. This study aimed to determine the effectiveness of tri-axial accelerometers deployed in a neck-collar and an ear-tag, to characterise the licking behaviour of beef cattle in individual pens. Four, 2-year-old Angus steers weighing 368 ± 9.3 kg (mean ± SD) were used in a 14-day study. Four machine learning (ML) algorithms (decision trees [DT], random forest [RF], support vector machine [SVM] and k-nearest neighbour [kNN]) were employed to develop behaviour classification models using three different ethograms: (1) licking vs. eating vs. standing vs. lying; (2) licking vs. eating vs. inactive; and (3) licking vs. non-licking. Activities were video-recorded from 1000 to 1600 h daily when access to supplement was provided. The RF algorithm exhibited a superior performance in all ethograms across the two deployment modes with an overall accuracy ranging from 88% to 98%. The neck-collar accelerometers had a better performance than the ear-tag accelerometers across all ethograms with sensitivity and positive predictive value (PPV) ranging from 95% to 99% and 91% to 96%, respectively. Overall, the tri-axial accelerometer was capable of identifying licking behaviour of beef cattle in a controlled environment. Further research is required to test the model under actual grazing conditions.

Abattoir Factors Influencing the Incidence of Dark Cutting in Australian Grain-Fed Beef.

The aim of this study was to evaluate the effect of carcass traits, lairage time and weather conditions during lairage and abattoir factors that impact the incidence of dark cutting in 142,228 grain-fed carcasses, as defined by Meat Standards Australia (MSA) guidelines. This study was conducted over a 12-month period analysing data from cattle that were supplied from seven feedlots and processed at three abattoirs. Abattoir data indicated that the average incidence of dark cutting within the study was 2.8%. Increased wind speeds (WSs) and rain during lairage at the abattoir was associated with an increased risk of dark cutting, whereas variation in ambient temperature and/or relative humidity did not influence dark cutting. Heavier carcasses with whiter fat, larger hump heights, more rib fat, higher marble scores and lower ossification had lower incidences of dark cutting. The factors abattoir, time in lairage, time to grading and grader within Abattoir had significant effects on the incidence of dark cutting. The results from this study suggest that reducing the time in lairage and increasing the time between slaughter and grading are the two major ways to reduce dark cutting in MSA carcasses.

The Influence of Predictability and Controllability on Stress Responses to the Aversive Component of a Virtual Fence.

To ensure animal welfare is not compromised, virtual fencing must be predictable and controllable, and this is achieved through associative learning. To assess the influence of predictability and controllability on physiological and behavioral responses to the aversive component of a virtual fence, two methods of training animals were compared. In the first method, positive punishment training involved sheep learning that after an audio stimulus, an electrical stimulus would follow only when they did not respond by stopping or turning at the virtual fence (predictable controllability). In the second method, classical conditioning was used to associate an audio stimulus with an electrical stimulus on all occasions (predictable uncontrollability). Eighty Merino ewes received one of the following treatments: control (no training and no stimuli in testing); positive punishment training with an audio stimulus in testing (PP); classical conditioning training with only an audio stimulus in testing (CC1); and classical conditioning training with an audio stimulus followed by electrical stimulus in testing (CC2). The stimuli were applied manually with an electronic collar. Training occurred on 4 consecutive days with one session per sheep per day. Sheep were then assessed for stress responses to the cues by measuring plasma cortisol, body temperature and behaviors. Predictable controllability (PP) sheep showed no differences in behavioral and physiological responses compared with the control treatment (P < 0.05). Predictable uncontrollability of receiving the aversive stimulus (CC2) induced a higher cortisol and body temperature response compared to the control but was not different to CC1 and PP treatments. CC2 treatment sheep showed a higher number of turning behaviors (P < 0.001), and more time spent running (P < 0.001) than the control and PP treatment groups, indicating that predictability without controllability was stressful. The behavior results also indicate that predicting the event without receiving it (CC1) was less stressful than predicting the event then receiving it (CC2), suggesting that there is a cost to confirmation of uncontrollability. These results demonstrate that a situation of predictability and controllability such as experienced when an animal successfully learns to avoid the aversive component of a virtual fence, induces a comparatively minimal stress response and does not compromise animal welfare.

Core and peripheral site measurement of body temperature in short wool sheep.

Understanding circadian rhythms of body temperature is important for the interpretation of single body temperature measurements and the assessment of the physiological state of an animal. The ability to measure body temperature at peripheral locations may also be important in the development of minimally invasive tools for remote temperature measurement in livestock. This study aimed to investigate how well body temperature measured at peripheral sites reflected a commonly used core measurement (vaginal temperature) and the circadian rhythmicity of the body temperature of sheep with a view to practical application in extensive sheep production systems. Eleven crossbred ewes were implanted with peripheral temperature sensing microchips (LifeChip®) which were positioned transversely in the sternocleidomastoid (neck) muscle and subcutaneously under the tail. iButton® temperature loggers were placed intravaginally to record core body temperature measurements (Tv). The body temperature measurements observed at the peripheral sites in the neck (Tn) and tail (Tt) differed significantly to those measured at the core site, Tv (P < 0.05), with Tn lower than Tv and Tt lower than both Tv and Tn. Similarities in circadian rhythm patterns were observed across the day between Tv, Tn and Tt in repeated measures analysis, with a short period of difference between Tv and Tn (from 1400 to 1600 h) and a long period of difference between Tv and Tt (from 1000 to 2100 h) (P < 0.05). These results suggest that neck muscle temperature measurements may have utility in detecting circadian rhythm patterns in core temperature in sheep, but may not accurately reflect absolute core temperatures. Peripheral measures may require adjustment or correction to more accurately reflect absolute core temperature with respect to determining accurate clinical thresholds relative to the expected normal temperature for the time of day observed. Further investigation into the utility and application of peripheral measurement of body temperature is warranted.

The Effect of Virtual Fencing Stimuli on Stress Responses and Behavior in Sheep.

To understand the animal welfare impact of virtual fencing stimuli (audio cue 'beep' and electrical stimulus) on naïve sheep, it is necessary to assess stress responses during the animal's first encounters with these stimuli. Eighty Merino ewes were exposed to one of the following treatments (n = 16 animals per treatment): Control (no stimuli), beep, dog bark, manual restraint, and electrical stimulus. Collars were used to apply the audio and electrical stimuli. The restraint treatment showed an elevated cortisol response compared with the control (p < 0.05), but there were no differences between the other treatments and the control. There were no differences between treatments in vaginal temperature (p > 0.05). For behaviors, the sheep receiving the bark and beep treatments were more vigilant compared to the control (p < 0.05), there were more aversive responses observed in the electrical stimulus treatment compared to the control. Together, the responses showed that the beep stimuli were largely benign, the bark stimuli was minimally aversive, the electrical stimuli was acutely aversive, and the restraint was moderately aversive. These data suggest that, for sheep, their first exposure to the virtual fencing stimuli should be perceived as less aversive than a commonly used restraint procedure.