Framework for integrating animal welfare into life cycle sustainability assessment

Purpose

This study seeks to provide a framework for integrating animal welfare as a fourth pillar into a life cycle sustainability assessment and presents three alternative animal welfare indicators.

Methods

Animal welfare is assessed during farm life and during slaughter. The indicators differ in how they value premature death. All three consider (1) the life quality of an animal such as space allowance, (2) the slaughter age either as life duration or life fraction, and (3) the number of animals affected for providing a product unit, e.g. 1 Mcal. One of the indicators additionally takes into account a moral value denoting their intelligence and self-awareness. The framework allows for comparisons across studies and products and for applications at large spatial scales. To illustrate the framework, eight products were analysed and compared: beef, pork, poultry, milk, eggs, salmon, shrimps, and, as a novel protein source, insects.

Results and discussion

Insects are granted to live longer fractions of their normal life spans, and their life quality is less compromised due to a lower assumed sentience. Still, they perform worst according to all three indicators, as their small body sizes only yield low product quantities. Therefore, we discourage from eating insects. In contrast, milk is the product that reduces animal welfare the least according to two of the three indicators and it performs relatively better than other animal products in most categories. The difference in animal welfare is mostly larger for different animal products than for different production systems of the same product. This implies that, besides less consumption of animal-based products, a shift to other animal products can significantly improve animal welfare.

Conclusions

While the animal welfare assessment is simplified, it allows for a direct integration into life cycle sustainability assessment. There is a trade-off between applicability and indicator complexity, but even a simple estimate of animal welfare is much better than ignoring the issue, as is the common practice in life cycle sustainability assessments. Future research should be directed towards elaborating the life quality criterion and extending the product coverage.

     

Transfer of passive immunity in dairy calves: the effectiveness of providing a supplementary colostrum meal in addition to nursing from the dam

Failed transfer of passive immunity (FTPI) in dairy calves – which is often due to the low amount of colostrum provided within a few hours after birth – remains a crucial issue. Enabling dairy calves to nurse colostrum from their dams could be useful in increasing intake and thus avoiding FTPI, but further potential effects on the health and welfare of both calves and dams should also be considered. In this study, 107 calf-dam pairs from two Italian dairy farms were alternately assigned to one of the following colostrum provision methods (CPMs): ‘hand-fed method’ (HFM) – the calf was separated from the dam immediately after birth and colostrum was provided by nipple-bottle (n = 50); ‘nursing method’ (NM) – the calf nursed colostrum from the dam for the first 12 h of life without farmer assistance (n = 30); and ‘mixed method’ (MM) – the nursing calf received a supplementary colostrum meal by nipple-bottle (n = 27). Serum of calves (1 to 5 days of age) and samples of their first colostrum meal were analysed by electrophoresis to assess immunoglobulin (Ig) concentration. Additionally, behavioural indicators of separation distress (calf and dam vocalisations; calf refusal of the first meal after separation; undesirable dam behaviour at milking) in the following 24 h were recorded as binary variables (Yes/No), and the health status of calves (disease occurrence and mortality) and dams (postpartum disorders and mastitis occurrence) were monitored for the first 3 months of life and 7 days after parturition, respectively. The lowest FTPI occurrence (calf serum Ig concentration <10.0 g/l) was found in the MM (11.1%) and the HFM (22.0%) compared with the NM (60.0%) (P<0.05), and the highest percentage of calves with optimal transfer of passive immunity (serum Ig concentration ≥16.0 g/l) was observed in the MM (55.6%). The lowest calf–dam separation distress was observed in the HFM (P<0.05). The highest calf disease occurrence was recorded in the HFM (64.0%) and the lowest in the NM (33.3%), with an intermediate value for the MM (44.4%) (P<0.05). No effect of the CPM was observed on dam health or calf mortality (P>0.05). The results of this study indicated that providing calves with a supplementary colostrum meal in addition to nursing from the dam (MM) is truly effective in maximizing passive immunity transfer. Anyway, specific strategies should be studied to minimise calf-dam separation distress.     

Design,synthesis and biological evaluation of novel 4,5-dihydro-[1,2,4]triazolo[4,3-f]pteridine derivatives as potential BRD4 inhibitors

Recently, diverse kinase inhibitors were reported having interaction with BRD4. It provided a strategy for developing a new structural framework for the next-generation BRD4-selective inhibitors. Starting from PLK1 kinase inhibitor BI-2536, we designed 18 compounds by modifying dihydropteridine core. Compound 23 showed potent BRD4 inhibitory activities with IC₅₀ of 79 nM and no inhibitory activities for PLK1. Cell antiproliferation assay was performed and potent inhibitory activity against MV4;11 with IC₅₀ of 1.53 μM. Cell apoptosis and western blotting indicated compound 23 induced apoptosis by down-regulating c-Myc. These novel selective BRD4 inhibitors provided new lead compounds for further drug development.     

Associations between behaviour and health outcomes in conventional and slow-growing breeds of broiler chicken

Broiler chickens are prone to a range of complex health and welfare issues. To support informed selection of welfare traits whilst minimising impact on production efficiency and to address a major gap in understanding, we systematically explored associations between health and behavioural indicators of broiler welfare. One conventional (CNV, n = 350) and two slow-growing broiler breeds (SGH and SGN, respectively n = 400) were reared from hatch in pens of 50 birds. Birds were assessed for health (gait, plumage cover and dirtiness, pododermatitis, hockburn, and leg deviations) at 2.2 kg liveweight according to the Royal Society for the Prevention of Cruelty to Animals Broiler Breed Welfare Assessment Protocol. Behaviour and resource-use of 10% of birds per pen, on days 29 (all breeds) and 43 (SGH and SGN), was (i) scan sampled every 60 min between three to six and between twelve to fifteen hours after photoperiod onset; and (ii) continuously sampled sequentially from focal birds for 3 min each in a random order, during 15 min observation periods at three and twelve hours after photoperiod onset. Binary logistic generalized linear models were used, to assess respective associations between pen prevalence of each health outcome and (i) pen mean percentage scans of behaviour, and (ii) pen mean frequency and duration per 3 min focal observation of behaviour. Better growth rate and feed conversion but poorer health outcomes (mortality, gait, pododermatitis, feather cover) were more prevalent in CNV. Strong associations between behaviour and several heath indicators revealed, (i) increases in side-lying inactive, sitting inactive, and use of the litter relative to other resources, as primary and general indicators of poorer health, and (ii) increases in standing inactive, perch use, walking, Comfort, High Energy and Exploratory behaviour as primary and general indicators of better health. Of these, changes in side-lying, standing inactive, walking, Comfort and High Energy behaviour were particularly sensitive to small differences in health outcomes important for breed acceptance in high-welfare schemes. Crucially these behavioural measures additionally represent motivational and affective aspects of welfare not captured by health measures and allow opportunity for earlier intervention. Thus, to provide a comprehensive assessment of broiler experience, behaviour should be incorporated into broiler welfare assessments.     

The effect of calf jacket usage on performance,behaviour and physiological responses of group-housed dairy calves

Poor performance and ill-health of calves in the pre-wean period can affect future productivity. Increasing numbers of producers are opting to use calf jackets as a means of mitigating the potential negative effects of low ambient temperatures, wind speed and precipitation on growth and health. This study aimed to use a range of noninvasive monitoring technologies to investigate the effects of using calf jackets in the first 3 weeks of life on calf performance and behavioural and physiological parameters. Ninety Holstein-Friesian calves were allocated to one of the two treatments: (i) Jacketed until 21 days of age and (J; n = 44) ii. Nonjacketed (NJ; n = 46). Calves were group housed and fed milk replacer (MR) and concentrate solid feed via automatic feeders. Calves were weaned at day 56, and the experiment was completed at day 63. Health assessments were conducted on a daily basis throughout the experiment using predefined faecal and respiratory scoring protocols. A range of novel, noninvasive monitoring technologies were used to examine the activity, heart rate and thermal profiles of calves on an individual basis throughout the experimental period. There were no differences in calf live weight (LWT), average daily gain (ADG) or feed conversion efficiency (FCE) in J and NJ calves between days 5 to 20. However, NJ calves consumed more MR and had more unrewarded visits to the milk feeder than J calves during this period. Although calf LWT was comparable across treatments in the week following jacket removal (days 21 to 28), both ADG and FCE tended to be greater in NJ calves. There were no treatment differences in calf LWT at the end of the study (d63). When measured over a period of 24 h and at a mean ambient temperature of 7.7°C, skin surface temperature was 6.37°C higher in J calves. Core body temperature was higher in J calves between days 5 to 20; however, there were no differences in IR eye or IR rectal temperature. No differences in lying behaviour occurred, with calves spending 18 and 17 h/day lying between days 5 to 20 and days 21 to 28, respectively. Under the climatic and management conditions described, no significant benefits to calf performance were found as a result of the provision of calf jackets to group-housed calves in the first 3 weeks of life. The higher frequency of unrewarded visits to the milk feeder in NJ calves during the first 3 weeks of life could be suggestive of a lack of satiety in these calves.     

Evaluation of on-farm veal calves’ responses to unfamiliar humans and potential influencing factors

The human–animal relationship is an important component of the welfare of farm animals and for this reason animal responsiveness tests to humans are included in on-farm welfare assessment schemes that provide indicators for this. However, apart from the behaviour of stockpersons towards their animals, other factors may also influence animals’ reactivity to humans as observed through behavioural tests, which can add a further layer of complexity to the interpretation of test results. Knowledge of these factors may help a better interpretation of differences from one farm to another in the outcome of human–animal relationship tests, and may provide clues for improving the relationship between animals and humans. The main objective of this study was to identify whether management or environmental factors could influence the outcome of human–animal relationship tests in veal calves. Two tests were performed when calves were aged 14.9 ± 1.6 (SD) weeks in 148 veal farms: the voluntary approach of an unfamiliar human standing at the feeding fence and the reaction towards an unfamiliar human who entered the home pen and tried to touch each calf in a standardised way (Calf Escape Test (CET) – score 0 to 4). Questionnaires were filled in and interviews with the stockpersons were performed in order to obtain information on stockpersons, management, animal and building characteristics. The latency to touch an unfamiliar human at the feeding fence was significantly correlated with the CET scores. Total number of calves on the farm, space allowance, breed, environmental enrichment, stockperson's experience and season of observation influenced the percentage of calves that scored 0 in CET (i.e. calves that could not be approached). Type of milk distribution, type of breed and number of calves per stockperson influenced the percentage of calves that scored 4 in CET (i.e. calves could be touched). For both CET0 and CET4, the level of self-reported contacts by the stockperson (analysed only on the French subset of 36 farms) did not influence the results. This paper concludes that according to the tests conducted on veal calves on commercial farms, factors such as milk distribution method, breed of the calves or the level of experience of stockpersons with veal farming can have an impact on the results of tests focusing on human–animal relationships.     

Indicators used in livestock to assess unconsciousness after stunning: a review

Assessing unconsciousness is important to safeguard animal welfare shortly after stunning at the slaughter plant. Indicators that can be visually evaluated are most often used when assessing unconsciousness, as they can be easily applied in slaughter plants. These indicators include reflexes originating from the brain stem (e.g. eye reflexes) or from the spinal cord (e.g. pedal reflex) and behavioural indicators such as loss of posture, vocalisations and rhythmic breathing. When physically stunning an animal, for example, captive bolt, most important indicators looked at are posture, righting reflex, rhythmic breathing and the corneal or palpebral reflex that should all be absent if the animal is unconscious. Spinal reflexes are difficult as a measure of unconsciousness with this type of stunning, as they may occur more vigorous. For stunning methods that do not physically destroy the brain, for example, electrical and gas stunning, most important indicators looked at are posture, righting reflex, natural blinking response, rhythmic breathing, vocalisations and focused eye movement that should all be absent if the animal is unconscious. Brain stem reflexes such as the cornea reflex are difficult as measures of unconsciousness in electrically stunned animals, as they may reflect residual brain stem activity and not necessarily consciousness. Under commercial conditions, none of the indicators mentioned above should be used as a single indicator to determine unconsciousness after stunning. Multiple indicators should be used to determine unconsciousness and sufficient time should be left for the animal to die following exsanguination before starting invasive dressing procedures such as scalding or skinning. The recording and subsequent assessment of brain activity, as presented in an electroencephalogram (EEG), is considered the most objective way to assess unconsciousness compared with reflexes and behavioural indicators, but is only applied in experimental set-ups. Studies performed in an experimental set-up have often looked at either the EEG or reflexes and behavioural indicators and there is a scarcity of studies that correlate these different readout parameters. It is recommended to study these correlations in more detail to investigate the validity of reflexes and behavioural indicators and to accurately determine the point in time at which the animal loses consciousness.     

Discovery of a new class of PROTAC BRD4 degraders based on a dihydroquinazolinone derivative and lenalidomide/pomalidomide

BRD4 has emerged as an attractive target for anticancer therapy. However, BRD4 inhibitors treatment leads to BRD4 protein accumulation, together with the reversible nature of inhibitors binding to BRD4, which may limit the efficacy of BRD4 inhibitors. To address these problems, a protein degradation strategy based on the proteolysis targeting chimera (PROTAC) technology has been developed to target BRD4 recently. Herein, we present our design, synthesis and biological evaluation of a new class of PROTAC BRD4 degraders, which were based on a potent dihydroquinazolinone-based BRD4 inhibitor compound 6 and lenalidomide/pomalidomide as ligand for E3 ligase cereblon. Gratifyingly, several compounds showed excellent inhibitory activity against BRD4, and high anti-proliferative potency against human monocyte lymphoma cell line THP-1. Especially, compound 21 (BRD4 BD1, IC₅₀ = 41.8 nM) achieved a submicromolar IC₅₀ value of 0.81 μM in inhibiting the growth of THP-1 cell line, and was 4 times more potent than compound 6. Moreover, the mechanism study established that 21 could effectively induce the degradation of BRD4 protein and suppression of c-Myc. All of these results suggested that 21 was an efficacious BRD4 degrader for further investigation.     

Influence of dietary strategy on progression of health and behaviour in mule ducks reared for fatty liver production

Overfeeding in ducks is questioned because forced introduction of food into the animal and metabolic overload may induce damage to health and discomfort. In this context, the objective of our experiment was to measure the impact of dietary strategy on the progression of animal status evaluated through 28 health and behavioural indicators in ducks reared for fatty liver production. To do this, 320 ducks were forced-fed twice a day from 70 to 90 days of age including 10 days of overfeeding (87 to 96 days). They were divided into two groups differing in the feeding strategy during overfeeding period: a moderate feed intake (MI, an average of 376 g of maize flour per meal, n = 160 ducks) or at high feed intake (HI, 414 g/meal, n = 160 ducks). We evaluated 28 indicators related to Good feeding (n = 3), Good housing (n = 4), Good health (n = 10) and Appropriate behaviour (n = 11) principles, taken from the European Welfare Quality Consortium® at four stages: before overfeeding (BEF; 80 days), at the beginning (88 days), the middle (MID; 92 days) or the end of overfeeding (END; 96 days). Animals were slaughtered at 93 and 97 days to measure fatty liver weight at MID and END stages (n = 80 per group). The results showed that dietary strategy influenced the fatty liver weight at MID (+23% in HI v. MI group; P < 0;05) and END stage (+23%; P < 0.05). Assessment stage influenced 13 of the 28 indicators measured. Among these 13 indicators, (i) BEF differed from END stage for 7 indicators and (ii) the dietary strategy degraded all the indicators chosen to evaluate the Good feeding (2/2) principle, but had no effect on the indicators related to the Good health (0/4) principle while (iii) most of the indicators that evaluated Good housing (2/3) and Appropriate behaviour (2/4) principles were affected by an interaction between both factors. Our results suggest that (i) duck fattening status, including the fatty liver weight, and several welfare indicators progressed during the fatty liver production process; and (ii) feeding strategy influenced or even interacted with this progression.     

Design,synthesis and biological evaluation of novel 6-phenyl-1,3a,4,10b-tetrahydro-2H-benzo[c]thiazolo[4,5-e]azepin-2-one derivatives as potential BRD4 inhibitors

Bromodomain-containing protein 4 (BRD4) is a key epigenetic regulator in cancer, and inhibitors targeting BRD4 exhibit great anticancer activity. By replacing the methyltriazole ring of the BRD4 inhibitor I-BET-762 with an N-methylthiazolidone heterocyclic ring, fifteen novel BRD4 inhibitors were designed and synthesized. Compound 13f had a hydrophobic acetylcyclopentanyl side chain, showing the most potent BRD4 inhibitory activity in the BRD4-BD1 inhibition assay (IC₅₀ value of 110 nM), it also significantly suppressed the proliferation of MV-4-11 cells with high BRD4 level (IC₅₀ value of 0.42 μM). Furthermore, the potent apoptosis-promoting and G0/G1 cycle-arresting activity of compound 13f were indicated by flow cytometry. As the downstream-protein of BRD4, c-Myc was in significantly low expression by compound 13f treatment in a dose-dependent manner. All the findings supported that this novel compound 13f provided a perspective for developing effective BRD4 inhibitors.     

Exploiting a water network to achieve enthalpy-driven,bromodomain-selective BET inhibitors

Within the last decade, the Bromodomain and Extra-Terminal domain family (BET) of proteins have emerged as promising drug targets in diverse clinical indications including oncology, auto-immune disease, heart failure, and male contraception. The BET family consists of four isoforms (BRD2, BRD3, BRD4, and BRDT/BRDT6) which are distinguished by the presence of two tandem bromodomains (BD1 and BD2) that independently recognize acetylated-lysine (KAc) residues and appear to have distinct biological roles. BET BD1 and BD2 bromodomains differ at five positions near the substrate binding pocket: the variation in the ZA channel induces different water networks nearby. We designed a set of congeneric 2- and 3-heteroaryl substituted tetrahydroquinolines (THQ) to differentially engage bound waters in the ZA channel with the goal of achieving bromodomain selectivity. SJ830599 (9) showed modest, but consistent, selectivity for BRD2-BD2. Using isothermal titration calorimetry, we showed that the binding of all THQ analogs in our study to either of the two bromodomains was enthalpy driven. Remarkably, the binding of 9 to BRD2-BD2 was marked by negative entropy and was entirely driven by enthalpy, consistent with significant restriction of conformational flexibility and/or engagement with bound waters. Co-crystallography studies confirmed that 9 did indeed stabilize a water-mediated hydrogen bond network. Finally, we report that 9 retained cytotoxicity against several pediatric cancer cell lines with EC₅₀ values comparable to BET inhibitor (BETi) clinical candidates.     

Scientists' Assessment of the Impact of Housing and Management on Animal Welfare

A total of 22 scientists contributed to a consensus-oriented conceptual framework for assessment of farm animal welfare, addressing priority issues in cattle, pigs, and chickens. They used the Delphi method, in which participants contributed anonymously through e-mail. The framework puts welfare in an evolutionary context. Welfare problems arise when animals are kept in environments to which they have difficulty adapting. For integrated welfare assessment, design criteria and welfare performance criteria must be used in which the former may be weighted based on the latter, which, in turn, may be weighted based on the aspects of intensity, duration, and incidence. The main performance criteria were pathological states, abnormal behaviors, and behavioral restrictions. Space, social conditions, and substrates were the main design criteria. Conventional housing systems generally ranked lower than alternative systems. In this article, we show that a broad consensus basis exists among welfare scientists concerning the assessment of farm animal welfare.     

Scientists' Assessment of the Impact of Housing and Management on Animal Welfare

A total of 22 scientists contributed to a consensus-oriented conceptual framework for assessment of farm animal welfare, addressing priority issues in cattle, pigs, and chickens. They used the Delphi method, in which participants contributed anonymously through e-mail. The framework puts welfare in an evolutionary context. Welfare problems arise when animals are kept in environments to which they have difficulty adapting. For integrated welfare assessment, design criteria and welfare performance criteria must be used in which the former may be weighted based on the latter, which, in turn, may be weighted based on the aspects of intensity, duration, and incidence. The main performance criteria were pathological states, abnormal behaviors, and behavioral restrictions. Space, social conditions, and substrates were the main design criteria. Conventional housing systems generally ranked lower than alternative systems. In this article, we show that a broad consensus basis exists among welfare scientists concerning the assessment of farm animal welfare.     

Stunning fish with CO2 or electricity: contradictory results on behavioural and physiological stress responses

Studies that address fish welfare before slaughter have concluded that many of the traditional systems used to stun fish including CO₂ narcosis are unacceptable as they cause avoidable stress before death. One system recommended as a better alternative is electrical stunning, however, the welfare aspects of this method are not yet fully understood. To assess welfare in aquaculture both behavioural and physiological measurements have been used, but few studies have examined the relationship between these variables. In an on-site study aversive behaviours and several physiological stress indicators, including plasma levels of cortisol and ions as well as blood physiological variables, were compared in Arctic char (Salvelinus alpinus) stunned with CO₂ or electricity. Exposure to water saturated with CO₂ triggered aversive struggling and escape responses for several minutes before immobilization, whereas in fish exposed to an electric current immobilization was close to instant. On average, it took 5 min for the fish to recover from electrical stunning, whereas fish stunned with CO₂ did not recover. Despite this, the electrically stunned fish had more than double the plasma levels of cortisol compared with fish stunned with CO₂. This result is surprising considering that the behavioural reactions were much more pronounced following CO₂ exposure. These contradictory results are discussed with regard to animal welfare and stress physiological responses. The present results emphasise the importance of using an integrative and interdisciplinary approach and to include both behavioural and physiological stress indicators in order to make accurate welfare assessments of fish in aquaculture.     

Animal performance and stress at weaning when dairy cows suckle their calves for short versus long daily durations

Calves in most dairy farms are separated from their dams either immediately or within a few hours after birth, prompting increasing concern of the society for reasons of animal welfare. The aim of this study was to identify systems to maintain cow-calf contact (CCC) that balance the benefits for calf growth and health against the negative impacts on sellable milk and stress at weaning. We tested reuniting cows and calves for 20 min before (Before-group) or 2.5 h after (After-group) morning milking (in Trial 1) or for a 9 h period between the morning and evening milkings (Half-day-group, in Trial 2). In Control-groups, calves were separated from their dam at birth and fed with artificial nipple with tank milk provided daily at 13% (Trial 1) and 14% (Trial 2) of their BW. In both trials, each practice was applied on a group of 14 dam–calf pairs (7 Holstein [Ho] and 7 Montbéliarde [Mo]). All calves were weaned at a BW of at least 100 kg. In Trial 1, the After-group was prematurely stopped when the calves were eight weeks of age as calf growth became limited (340 g/d) due to low milk intakes (2.97 kg/d). During the first eight weeks of lactation, milk yield at the parlour was 29%, 51% and 42% lower in After-, Before- and Half-day-cows respectively compared to Controls. From week 14 to 16 when all calves were separated from their dam, Before-cows still produced 25% less milk than Control-cows while Half-day-cows reached the milk yield of Control-cows within a week. There were no significant differences in milk somatic cell count and in frequency of health disorders (cows and calves) between suckling and Control-groups. Compared to Control-calves, calf growth until weaning was higher in the suckling calves in Trial 1 (861 vs 699 g/d) and similar in Trial 2 (943 vs 929 g/d). At weaning, Before- and Half-day-calves started to vocalise earlier and continued to vocalise longer than Controls. In conclusion, the best compromise between cow milk yield and calf growth is a long period of CCC (9 h) between the morning and evening milkings. Still abrupt weaning stresses both cows and calves even if CCC has been restricted before separation.