C. R. Ralph

INVITED REVIEW: The usefulness of measuring glucocorticoids for assessing animal welfare

Glucocorticoids (corticosterone in birds and rodents and cortisol in all other mammals) are glucoregulatory hormones that are synthesized in response to a range of stimuli including stress and are regularly measured in the assessment of animal welfare. Glucocorticoids have many normal or non-stress-related functions, and glucocorticoid synthesis can increase in response to pleasure, excitement, and arousal as well as fear, anxiety, and pain. Often, when assessing animal welfare, little consideration is given to normal non-stress-related glucocorticoid functions or the complex mechanisms that regulate the effects of glucocorticoids on physiology. In addition, it is rarely acknowledged that increased glucocorticoid synthesis can indicate positive welfare states or that a stress response can increase fitness and improve the welfare of an animal. In this paper, we review how and when glucocorticoid synthesis increases, the actions mediated through type I and type II glucocorticoid receptors, the importance of corticosteroid-binding globulin, the role of 11 β-hydroxysteroid dehydrogenase, and the key aspects of neurophysiology relevant to activating the hypothalamo-pituitary-adrenal axis. This is discussed in the context of animal welfare assessment, particularly under the biological functioning and affective states frameworks. We contend that extending the assessment of animal welfare to key brain regions afferent to the hypothalamus and incorporating the aspects of glucocorticoid physiology that affect change in target tissue will advance animal welfare science and inspire more comprehensive assessment of the welfare of animals.

Impact of psychosocial stress on gonadotrophins and sexual behaviour in females: role for cortisol?

This review focuses on the importance of cortisol in mediating the inhibitory effects of psychosocial stress on reproduction in females. In particular, we have summarized our research in sheep where we have systematically established whether cortisol is both sufficient and necessary to suppress reproductive hormone secretion and inhibit sexual behaviour. Our findings are put into context with previous work and are used to develop important concepts as well as to identify productive further lines of investigation. It is clear that cortisol is necessary to inhibit some, but not all, aspects of reproduction in female sheep. These actions vary with reproductive state, and there are important interactions with gonadal steroids. The impact of cortisol on the tonic secretion of gonadotrophin-releasing hormone and luteinizing hormone has been investigated extensively, but less is known about the surge secretion of these hormones and their effects on sexual behaviour. Furthermore, there are separate effects of cortisol in the brain (hypothalamus) and at the anterior pituitary, illustrating that there are different mechanisms of action. Thus, although cortisol is important in mediating some of the effects of stress on reproduction, we need to look beyond cortisol and investigate some of the other mechanisms and mediators that relay the effects of stress on reproduction. In this regard, we propose that a group of neurons in the hypothalamus that co-synthesize kisspeptin, neurokinin B and dynorphin, termed KNDy cells, play important roles in mediating the effects of cortisol on reproduction. This hypothesis needs to be rigorously tested.

The hypothalamo-pituitary-adrenal (HPA) axis in sheep is attenuated during lactation in response to psychosocial and predator stress

Activation of the hypothalamo-pituitary-adrenal (HPA) axis by psychosocial stress is attenuated during lactation. We tested the hypothesis that lactating ewes will have attenuated HPA axis responses to isolation and restraint but will have greater responses to predator stress in the form of barking dogs. We imposed two 4 h stressors: psychosocial stress (isolation and restraint of ewes) and predator stress (barking dogs). Blood was collected intravenous every 10 min from nonlactating ewes (n = 6), lactating ewes with lambs present but not able to be suckled (n = 6), and lactating ewes with lambs present and able to be suckled (n = 6). Plasma cortisol and oxytocin were measured. For nonlactating ewes, cortisol increased (P < 0.01) in response to both stressors, and these increases were greater (P < 0.01) than that in the lactating animals. For lactating ewes with lambs present but unable to be suckled, cortisol increased (P < 0.05) in response to both stressors with a greater response to barking dogs (P < 0.05). For lactating ewes with lambs present and able to be suckled, cortisol increased (P < 0.01) in response to barking dogs only. Plasma oxytocin was greater (P < 0.01) in lactating ewes than in nonlactating ewes and did not change in response to the stressors. In conclusion, lactating ewes are likely to have a greater HPA axis response to a stressor that may be perceived to threaten the welfare of themselves and/or their offspring. The role of oxytocin in attenuation of the HPA axis to stress in sheep is unclear from the current research and requires further investigation.

Response to gonadotrophins differs for gilts from female- and male-biased litters

In several species, females masculinised by abnormal androgen exposure in utero have poor reproductive performance and gilts born into litters with a male bias are likely exposed to greater androgen concentrations prenatally than gilts born into female-biased litters. At 24h of age, piglet plasma testosterone concentrations in gilts from male-biased litters (>60% male; n=22) or female-biased litters (>60% female; n=27) were not different. At 18 wks of age, all gilts received an injection of 400IU equine chorionic gonadotrophin plus 200IU human chorionic gonadotrophin to stimulate oestrus. Two weeks after the injection gilts were slaughtered and ovaries collected for determination of numbers of corpora lutea (CL). Compared to gilts from female-biased litters, gilts from male-biased litters were more likely to ovulate (86.0% vs 59.5%, P=0.047) and had more CL (13.1±1.5 vs 7.2±1.7, P=0.015). The present data indicate an effect of birth litter sex-bias on pre-pubertal physiological development, possibly involving organisational effects at the ovarian cellular level impacting on future ovarian function. Potential impacts on subsequent fertility remain to be determined.

Hormones, stress and the welfare of animals

There are numerous endocrine (hormonal) responses during stress and these are often complex. This complexity makes the study of endocrine stress responses challenging and the challenges are intensified when attempts are made to use measures of hormones to assess the welfare of animals because so many endocrine systems are activated during stress and because there are countless stimuli that trigger these systems. Most research has concentrated on only a small number of these endocrine systems, particularly the hypothalamo–pituitary adrenal axis and the sympathoadrenal system, and there is a need to broaden the scope of endocrine systems that are studied. Furthermore, systematic approaches are required to establish when the actions of hormones associated with stress responses result in physiological and/or behavioural consequences that will have negative or positive effects on the welfare of animals.

Neurophysiological assessment of animal welfare

Livestock industries such as the pork industry are striving to continuously improve the welfare of animals. Inherent to the success of this is the ability to rigorously assess the welfare of animals in the field. While much progress has been made towards the development of methodology to assess the welfare of animals, there have been major challenges to establishing practical and definitive procedures to assess the welfare of animals. These include, but are not limited to, establishing a universally accepted definition of animal welfare and the choice of measures that are taken from the animal to assess its welfare. Measures of biological functioning and affective (emotional) state of the animal have been common, but there have been many limitations in terms of practical application. Some of the reasons for this include the choice of physiological measures, which are often restrictive in providing information about welfare, affective measures being restricted to specific behavioural measures and the biological-functioning and affective-states approaches being undertaken in isolation. Biological and affective functioning are integrated and controlled by the brain. Many of the regions of the brain involved in the regulation of biological and emotional functioning have been identified. Furthermore, there is considerable knowledge about the roles and interactions among the neurophysiological systems in these brain regions. We propose a strategy to use this knowledge to develop procedures to assess animal welfare. The initial phase is to identify the neural pathways that regulate the physiological and emotional processes that allow animals to adapt and cope. The next phase is to determine the activity of these pathways in conscious animals in the field. This requires the identification of biomarkers of specific neuronal activity that can be measured in the conscious animal in the field. Emerging technologies are offering promise in the identification of such biomarkers and some of these are already applicable to the pig. There is now the opportunity to apply this strategy within the pork industry to assess the welfare of pigs throughout the value chain.

Environmental Enrichment for Sucker and Weaner Pigs: The Effect of Enrichment Block Shape on the Behavioural Interaction by Pigs with the Blocks

Simple Summary How often do intensively-housed pigs interact with enrichment devices, especially smaller pigs during the sucker and weaner phases of production? Understanding whether smaller pigs use such devices, whether habituation occurs, and whether device-shape influences the level of interaction by pigs are all relevant questions surrounding justification for including enrichment devices in pig pens. We provided litters of pigs from 10 to 60 days old with one of three different shaped enrichment blocks (cube, brick, or wedge) and recorded oro-nasal contact with the blocks on two days each week. Fresh blocks were provided weekly. While pig interaction with blocks was infrequent before about 25 days of age, contact with the blocks steadily increased thereafter. Brick-shaped blocks were used more than the other shapes, possibly because the brick-shape presented a wider surface for contact, allowing multiple pigs to simultaneously interact with the block. Pigs interacted more with 1-day-old (i.e., ‘fresh’) than 4-day-old blocks, suggesting that habituation to the blocks may have occurred. Abstract This experiment tested the effect of enrichment-block shape on oro-nasal contact by young pigs, and possible habituation to the blocks. Nineteen litters (197 piglets) were randomly allocated to one of three block-shape treatments: Cube, Brick, or Wedge. Oro-nasal contact with blocks was infrequent before 25 days of age. Thereafter, contact steadily increased, suggesting enrichment blocks may not need to be provided until week 4 of lactation. Brick-shaped blocks attracted more oro-nasal contact than the cube and wedge shapes (p = 0.002). Oro-nasal contact was more frequent (p < 0.001) during the first 24 h after block introduction than when blocks were four days old. From 25 to 60 days of age, oro-nasal bouts were longer (p = 0.014) during the first 30 min of exposure to a fresh block, than for the remainder of the 24 h, or on day 4 after block replacement. Therefore, habituation to blocks may have occurred by 24 h after block introduction. Brick-shaped blocks may present a wider surface for oro-nasal contact, where multiple pigs could simultaneously interact with the block. We speculate that simultaneous interaction with brick-shaped blocks may be similar to a litter co-operatively massaging the sow’s udder prior to suckling bouts.

Sex bias of the birth litter affects surge but not tonic LH secretion in gilts

The physiology and behavior of gilts that develop in a male-biased litter can differ from gilts that develop in a female-biased litter. We hypothesized that gilts from male-biased litters will have a delayed and attenuated luteinizing hormone (LH) surge, and reduced LH pulse frequency and amplitude compared to gilts from female-biased litters. Gilts were selected at birth from male-biased (>60% males n = 10) or female-biased (>60% females n = 9) litters. From 18 wk of age, detection of puberty using daily boar contact began and their subsequent estrous periods were synchronized with oral progestogen (altrenogest). On day 3 after altrenogest withdrawal, blood samples were obtained from 6 gilts per sex bias group at 10 min intervals from 0900 to 2100 h to determine LH pulse amplitude and frequency. From 0900 on day 4, all 19 gilts were sampled every 4 h until the end of estrus to characterize LH surge dynamics. There were no differences between groups in LH pulse characteristics. Compared to gilts from female-biased litters, the LH surge in gilts from male-biased litters was delayed [56.00 ± 3.32 h vs. 43.11 ± 3.76 h (mean ± standard error of the mean (SEM)), P < 0.05], the duration was decreased [29.78 ± 2.12 h vs. 37.71 ± 1.19 h (mean ± SEM), P < 0.05] and the total secretion as measured by area under the curve was decreased (91.42 ± 9.52 ng/mL vs. 120.28 ± 9.48 ng/mL, P < 0.05). Our results indicate that a male-biased uterine environment has different effects on the tonic secretion of LH than the LH surge, with only some elements of the LH surge being affected.

Anogenital distance reflects the sex ratio of a gilt’s birth litter and predicts her reproductive success1

Anogenital distance (AGD) has been used to reflect masculinization in litter-bearing species. As masculinization affects behavior and reproduction, AGD could be measured to assist in selecting gilts with a temperament more suited to commercial production and greater reproductive potential. We hypothesized that gilts from a male-biased litter would have a longer AGD and poorer reproductive performance. In Exp. 1, AGD and weight were measured at day 1, day 21, and week 16 of age for gilts from male-biased litters (≥60% males; n = 51) and female-biased litters (≥60% females; n = 51). Sow AGD was measured 3 d after farrowing. In Exp. 2, AGD was measured at gilt selection at approximately 24 wk of age and gilts followed to second parity. Litter sex ratio affected AGD at 16 wk of age, with gilts from female-biased litters having longer AGD (mean ± SEM, 9.1 ± 0.7 mm vs. 11.0 ± 0.6 mm, P = 0.013). Anogenital distance was not different on day 1 or day 21. There was no effect of sex ratio on weight at any time, and sow AGD was not associated with the sex ratio of her litter. Gilts with an AGD longer than the mean of 11.55 mm were heavier (mean ± SEM, 118.8 ± 0.4 kg vs. 117.7 ± 0.4 kg, P = 0.023), were achieved puberty earlier (179.6 ± 0.6d vs. 182.2 ± 0.6 d, P = 0.001), were mated younger (200.6 ± 0.6 d vs. 203.2 ± 0.6 d, P = 0.001), and were more likely to be mated (91% vs. 83%, P = 0.005) than gilts with an AGD shorter than the mean. Gilts with an AGD greater than 11.55 mm had a greater born alive litter size (11.79 ± 0.20 vs. 11.20 ± 0.19, P = 0.018) compared with gilts with an AGD shorter than 11.55 mm. At 16 wk, AGD was associated with sex bias and could be used as a selection tool to predict reproductive success of the first parity, with a longer AGD being associated with gilts that had been born into a female-biased litter and that had better reproductive performance.

The sex ratio of a gilt’s birth litter can affect her fitness as a breeding female

Lifetime reproductive performance and behaviour are established in utero driven by genetics and the steroidal milieu of the uterine environment. Developing males synthesise androgens that can be absorbed by females in the litter. Consequently, the proportion of males in a litter has the potential to affect both lifetime reproductive performance and behaviour of female littermates. Although reproductive potential is understandably important for gilt selection, behaviour is becoming progressively more important as group housing of sows increases. Aggression can be a cause for removal from the breeding herd, and can also negatively affect reproductive performance and herd longevity. This review covers existing evidence of masculinisation in rodents and sheep, the mechanism causing masculinisation, and the knowledge gaps surrounding masculinisation in pigs. Premature culling of gilts is an economic and animal welfare issue in the pork industry resulting in low retention of gilts and reduced productivity. The selection of gilts needs to be improved to incorporate both lifetime reproductive potential and behaviour. We determined that there is cause to investigate the effect of the birth litter sex ratio of a gilt on her suitability for selection into the breeding herd.