Can a gradual weaning and separation process reduce weaning distress in dam-reared dairy calves? A comparison with the two-step method.

The weaning and separation phase remains one of the biggest challenges for cow-calf-contact systems, but a gradual process that better mimics the naturally occurring reduction in milk intake has not yet been scientifically investigated. Therefore, the aim of our study was to compare behavioral and physiological indicators of distress in 3 mo-old dam-reared dairy calves (with previous full-time cow-calf contact) weaned and separated either via gradual reduction of contact time with the dam (GR, 1 week half day contact, 1 week morning contact, 1 week fence-line contact before complete separation, n = 18) or via 2-step weaning using a nose flap (NF, 2 weeks access to dam with a nose flap, 1 week fence-line contact before complete separation, n = 18). Behavior was recorded 1 week before (or for lying 3 weeks before) weaning start and during the 3 week weaning and separation period with direct observations on 4 d per week or via accelerometers (locomotor play, lying behavior). Blood and fecal samples were taken twice per week from weaning start until 3 weeks after weaning start. Calves were weighed weekly. Statistical analysis was conducted using (generalized) linear mixed models. Over the whole weaning and separation phase, NF calves showed a stronger decrease in the number of lying bouts, amount of locomotor play and average daily weight gains, as well as a higher increase in total mixed ration feeding time compared with GR calves, whereas GR calves vocalized more often and showed more searching behavior than NF calves. Also, the neutrophil:lymphocyte ratio of NF calves was elevated on d 3 after insertion of the nose flaps compared with baseline, but showed no change for GR calves on any sampling day. Overall, results point toward a favorable effect of a gradual weaning strategy on reduction of weaning and separation distress in dam-reared dairy calves, but the method requires further improvement from the protocol used in our study.

Development and evaluation of an animal health and welfare monitoring system for veterinary supervision of pullet farms.

Regular welfare monitoring throughout rearing of pullets may help to identify problems early and take counteractions timely, which helps in guaranteeing good welfare. The aims of our observational study were (i) to establish and test a welfare monitoring system that can be used during (short) routine veterinary and technical staff visits for pullet flocks, (ii) to use the monitoring system to investigate variability between flocks and (iii) to analyse factors that potentially affect pullets' body weight, uniformity in body weight and mortality. The developed monitoring system tries to minimise the time required while not losing important information. Age-specific recording sheets comprise animal-based indicators of welfare and relevant environmental factors (housing, management, care) to allow for identifying causes of problems and targeted action. Finally, the system was implemented in a cross-sectional study and data collected in 100 flocks (67 organic, 33 conventional) on 28 rearing farms in Austria. Linear mixed models were used to identify factors influencing body weight, uniformity and mortality, both including all flocks (A) and only organic flocks (O) and a linear regression model with all flocks to investigate associations within animal-based indicators. High variability was found between flocks in animal-based indicators. Body weight was higher when the pre-rearing period was shorter (p ≤ 0.001, A&O), with higher intensities of light (p = 0.012, O), with only one compared to more stockpersons (p ≤ 0.007, A&O), with a higher number of flock visits per day (p ≤ 0.018, A&O), and a lower avoidance distance (p = 0.034, A). Body weight uniformity increased, with age and decreased with the duration of the light period (p = 0.046, A), and, amongst others, was higher on organic farms (farming type; p = 0.041). The latter may reflect a more uniform level of welfare due to a lower stocking density and lowered effects of social competition. Within organic flocks mortality was lower if pullets had access to a covered veranda (p = 0.025) resulting in an overall lower stocking density inside the barn, while in the model including all farms mortality was higher in cases where a disease had been diagnosed. We conclude that our monitoring system can easily be implemented in regular veterinary and technical staff visits, but could also be used by the farmers'. Several easy-to-record animal-based indicators of animal welfare could be analysed more frequently to increase early detection of problems. Implementation of such a routine-based monitoring system with easy-to-assess animal-based parameters and input measures can contribute to better animal health and welfare in pullets.

Welfare of calves.

This Scientific Opinion addresses a European Commission request on the welfare of calves as part of the Farm to Fork strategy. EFSA was asked to provide a description of common husbandry systems and related welfare consequences, as well as measures to prevent or mitigate the hazards leading to them. In addition, recommendations on three specific issues were requested: welfare of calves reared for white veal (space, group housing, requirements of iron and fibre); risk of limited cow-calf contact; and animal-based measures (ABMs) to monitor on-farm welfare in slaughterhouses. The methodology developed by EFSA to address similar requests was followed. Fifteen highly relevant welfare consequences were identified, with respiratory disorders, inability to perform exploratory or foraging behaviour, gastroenteric disorders and group stress being the most frequent across husbandry systems. Recommendations to improve the welfare of calves include increasing space allowance, keeping calves in stable groups from an early age, ensuring good colostrum management and increasing the amounts of milk fed to dairy calves. In addition, calves should be provided with deformable lying surfaces, water via an open surface and long-cut roughage in racks. Regarding specific recommendations for veal systems, calves should be kept in small groups (2-7 animals) within the first week of life, provided with ~ 20 m2/calf and fed on average 1 kg neutral detergent fibre (NDF) per day, preferably using long-cut hay. Recommendations on cow-calf contact include keeping the calf with the dam for a minimum of 1 day post-partum. Longer contact should progressively be implemented, but research is needed to guide this implementation in practice. The ABMs body condition, carcass condemnations, abomasal lesions, lung lesions, carcass colour and bursa swelling may be collected in slaughterhouses to monitor on-farm welfare but should be complemented with behavioural ABMs collected on farm.

Fecal cortisol metabolites reflect transport stress in 3-month-old dairy calves pre- and postweaning: A pilot study.

Measurement of fecal cortisol metabolites (FGCM) is a well-established, noninvasive method to assess stress in adult dairy cattle. However, this procedure has not yet been validated for unweaned dairy calves, and it can be expected that the milk proportion of the diet may influence the resulting FGCM concentrations. The aim of this study was therefore to assess whether a peak in FGCM concentrations in response to a stressor can be measured in unweaned dairy calves on a largely milk-based diet. If so, further objectives were to examine whether maximum FGCM concentrations, as well as the time lag until they are reached, are comparable to the values in the same calves on a solid-based diet after weaning. For this study, 5 German Holstein calves of about 3 mo of age (93 to 102 d preweaning) were exposed to a 45 min transport stressor once before and once after weaning, which was 3 wk apart. All voided fecal samples were collected for 24 h after termination of the transport. Fecal cortisol metabolites were analyzed with an 11-oxoetiocholanolone enzyme immunoassay and changes in FGCM concentrations relative to the individual baseline (FGCMrel) were calculated. Results showed a clear peak in FGCM concentrations on both diet types. The peak FGCMrel concentrations tended to be higher when the calves were on the preweaning diet (at peak: +233 ± 25% increase relative to baseline) in comparison to the postweaning diet (+124 ± 23%). Considering the whole 24 h sampling period, the FGCMrel concentrations for all calves were significantly higher on the preweaning diet than on the postweaning diet. There was also a numerical difference in the delay between occurrence of the stressor and appearance of the peak FGCMrel concentrations in feces, as the time lag was 1.5 ± 1.2 h longer when the calves were on the preweaning diet compared with the postweaning diet. In conclusion, our results suggest that FGCM concentrations are a useful stress marker for unweaned dairy calves in the same way they are for older cattle, but that FGCMrel concentrations tend to be higher in unweaned than in weaned calves and are thus not directly comparable.

Maternal deprivation affects goat kids' social behavior before and after weaning.

Maternal deprivation has been shown to disrupt the development of neonates. Nevertheless, separating the young animals from their dams soon after birth is a common practice in dairy farming. We investigated the effects of maternal deprivation on goat kids' (Capra hircus) social behavior and social ontogeny before and after weaning. Twenty female kids were raised together with their dams (DR kids) and other lactating goats and kids, whereas 20 female kids were separated from their dams 3 days after birth and artificially reared together (AR kids). At weaning, each treatment group was split in half and moved into two new pens where they were mixed with the other treatment group. Social behaviors were recorded before and after weaning. Before weaning, AR kids were observed performing more play-fighting, racing, stepping on each other, and standing in contact with each other than DR kids, but AR allogroomed less and spent less time resting alone than DR kids. After weaning and mixing of the treatments, DR kids initiated more and received less agonistic interactions than AR kids, but this difference reduced across the 5 weeks of observations as AR kids appeared to progressively change their social behavior after interacting with DR kids.

Maternal deprivation affects goat kids' stress coping behaviour.

Maternal deprivation early in life has been shown to disrupt neonates' development. Nevertheless, separating the young animals from their dams soon after birth remains a common practice in dairy farm husbandry. This study investigated the effects of different rearing conditions on goat kids' stress coping abilities. Twenty female kids were raised together with their dams ('dam-reared') in a herd composed of other lactating goats and kids, while twenty female kids were separated from their dams three days after birth and reared together with same-age peers ('artificially-reared') and visually separated from the lactating herd. All kids shared the same father and two thirds of the kids were twins allocated to each treatment. At one month of age, kids were individually submitted to a series of tests: a novel arena test, a novel goat test, and a novel object test. These tests happened consecutively in this order, and lasted 180 s each. The kids' behaviour was video-recorded and analysed post-hoc by an observer blind to treatments. Five weeks after weaning, the kids were also subjected to human-animal relationship tests. During the three behavioural tests, artificially-reared kids vocalized more (P < 0.001), reared more (P < 0.001), ran more (P = 0.002) and jumped more (P < 0.001) than dam-reared kids, but self-groomed less (P = 0.01) and urinated less (P = 0.05) than dam-reared kids. During the novel goat test and the novel object test, artificially-reared kids gazed less at the novel goat and the novel object (P = 0.02) and initiated contact more quickly (P = 0.05) with the novel goat and the novel object than dam-reared kids. The treatments however did not differ significantly in salivary cortisol response to the tests (P = 0.96). Artificially-reared kids showed significantly less avoidance of humans than dam-reared kids during the human-animal relationship tests after weaning (P < 0.001). The higher intensity of their behavioural reaction showed that artificially-reared kids react to stressful situations more actively than dam-reared kids. The difference between the three tests were only minor, suggesting a general change in the kids' response to stressful situations rather than a specific change in their social response tested with an unfamiliar adult. Hence, artificial rearing affects goat kids' behavioural response to challenges, probably maternal deprivation being the main factor.

Mechanical Nociceptive Threshold, Tissue Alterations and Horn Growth in Calves after Injection of Isoeugenol or Clove Oil under the Horn Bud.

Disbudding of calves is a common, painful intervention. Due to cytotoxic and anesthetic properties, the injection of clove oil or its component isoeugenol may be less detrimental to animal welfare. We investigated mechanical nociceptive threshold (MNT), possible tissue alterations and horn growth for up to 12 weeks after injection of 1.5 mL clove oil (CLOV), isoeugenol (ISO) or saline (CON) or after hot-iron disbudding (BURN; with local anesthesia and sedation, n = 10/treatment). MNT was measured using von Frey filaments and a pressure algometer at four locations around the horn bud. There was a treatment*time point interaction (linear mixed model, p < 0.05). MNT decreased most strongly and for the longest time for BURN in most calves at least for 3 weeks. For ISO, the decrease was less distinct and most calves' values returned to baseline after 1-2 weeks. MNT in CLOV was intermediate, with decreased values up to 3 weeks in some animals. 12 weeks after the treatment, horn growth was prevented in about 50% of the horns in CLOV and ISO. Tissue alterations such as swellings of the eyelids often occurred in CLOV, but less so in ISO. Our results suggest that injection of isoeugenol causes less pain and thus seems to be beneficial compared to hot-iron disbudding, while clove oil was not advantageous. Regarding the effectiveness of isoeugenol to prevent horn growth, more studies are needed.

Characteristics and Welfare of Long-Term Shelter Dogs.

To identify characteristics that distinguish long-term (LT: stay > 1 year) from short-term shelter dogs (ST: ≤5 months) and to investigate if a long-term stay impairs welfare, we compared ST and LT dogs in Austrian no-kill shelters. Analyses including characteristics such as breed, sex, or age (shelter records), problem behaviour, and personality (questionnaires completed by staff) showed that LT dogs were significantly more often a "dangerous breed", male, and older when admitted to the shelter. They were rated higher on "aggression" and "high arousal" and lower on the personality dimension "amicability". A welfare assessment protocol including reaction toward humans (Shelter Quality Protocol), and in-kennel observations were used to assess the effect of the long-term stay. LT dogs tended to show more signs of aggression toward an unfamiliar human, but welfare assessment revealed no difference. During resting periods, LT dogs spent more time resting head up and had more bouts resting head down. Prior to feeding, they stood, vocalised, and yawned more. LT dogs are characterised by specific features such as being aroused easily and having difficulties to relax. Whether this is a result of the long-term stay or personality-associated, consequently causing lower adoption rates, remains to be determined.

Gentle interactions with restrained and free-moving cows: Effects on the improvement of the animal-human relationship.

The animal-human relationship is essential for farm animal welfare and production. Generally, gentle tactile and vocal interactions improve the animal-human relationship in cattle. However, cows that are fearful of humans avoid their close presence and touch; thus, the animal-human relationship first has to be improved to a point where the animals accept stroking before their perception of the interactions and consequently the animal-human relationship can become positive. We tested whether the animal-human relationship of cows fearful of humans is improved more effectively by gentle interactions during restraint, allowing physical contact from the beginning, or if the gentle interactions are offered while the animals are free to move, giving them more control over the situation and thus probably a higher level of agency and a more positive perception of the interactions. Thirty-six dairy cows (median avoidance distance 1.6 m) were assigned to three treatments (each n = 12): gentle vocal and tactile interactions during restraint in the feeding rack (LOCK); gentle vocal and, if possible, tactile interactions while free in the barn (FREE); routine management without additional interactions (CON). Treatments were applied for 3 min per cow on 10 d per fortnight for 6 weeks (i.e., three periods). Avoidance and approach behaviour towards humans was tested before the start of the treatment period, and then at 2-week intervals. The recorded variables were reduced to one score by Principal Component Analysis. The resulting relationship score (higher values implying a better relationship with humans) increased in all groups; the increase was stronger in FREE than in CON, with the increase in LOCK being not significantly different from the other treatment groups. Thus, we recommend that gentle interactions with cows should take place while they are unrestrained, if possible.

The Power of a Positive Human-Animal Relationship for Animal Welfare.

Domestic animals often seek and enjoy interacting with humans. Positive human-animal relationships can elicit positive emotions and other positive welfare outcomes. Nevertheless, our understanding of the underlying processes that govern the positive perception of humans by animals is incomplete. We cover the potential mechanisms involved in the development and maintenance of positive human-animal relationships from the perspective of the animal. This encompasses habituation, associative learning, and possibly attachment or bonding based on communication and social cognition. We review the indicators from the literature to assess a positive human-animal relationship. We operationally define this positive relationship as the animal showing voluntary approach and spatial proximity (seeking) and signs of anticipation, pleasure, relaxation, or other indicators of a rewarding experience from interacting with the human. For research, we recommend accounting for the baseline human-animal relationship in the animal's everyday life, and incorporating a control treatment rather than only comparing positive to negative interaction treatments. Furthermore, animal characteristics, such as previous experience, genetics, and individual predisposition, as well as contextual characteristics related to the social and physical environment, may modulate the perception of humans by animals. The human-animal relationship is also influenced by human characteristics, such as the person's familiarity to the animal, attitudes, skills, and knowledge. We highlight implications for current practices and suggest simple solutions, such as paying attention to the animal's behavioral response to humans and providing choice and control to the animal in terms of when and how to interact with humans. Practical applications to achieve a positive perception of humans could be better utilized, such as by incorporating training principles, while keeping in mind trust and safety of both partners. Overall, there is growing evidence in the scientific literature that a positive human-animal relationship can bring intrinsic rewards to the animals and thereby benefit animal welfare. Further research is needed on the underlying processes to establish an effective positive human-animal relationship, especially in regard to the type, frequency, and length of human interaction necessary. In particular, the importance of providing animals with a sense of agency over their interactions with humans remains poorly understood.

Talking to Cows: Reactions to Different Auditory Stimuli During Gentle Human-Animal Interactions.

The quality of the animal-human relationship and, consequently, the welfare of animals can be improved by gentle interactions such as stroking and talking. The perception of different stimuli during these interactions likely plays a key role in their emotional experience, but studies are scarce. During experiments, the standardization of verbal stimuli could be increased by using a recording. However, the use of a playback might influence the perception differently than "live" talking, which is closer to on-farm practice. Thus, we compared heifers' (n = 28) reactions to stroking while an experimenter was talking soothingly ("live") or while a recording of the experimenter talking soothingly was played ("playback"). Each animal was tested three times per condition and each trial comprised three phases: pre-stimulus, stimulus (stroking and talking) and post-stimulus. In both conditions, similar phrases with positive content were spoken calmly, using long low-pitched vowels. All tests were video recorded and analyzed for behaviors associated with different affective states. Effects on the heifers' cardiac parameters were assessed using analysis of heart rate variability. Independently of the auditory stimuli, longer durations of neck stretching occurred during stroking, supporting our hypothesis of a positive perception of stroking. Observation of ear positions revealed longer durations of the "back up" position and less ear flicking and changes of ear positions during stroking. The predicted decrease in HR during stroking was not confirmed; instead we found a slightly increased mean HR during stroking with a subsequent decrease in HR, which was stronger after stroking with live talking. In combination with differences in HRV parameters, our findings suggest that live talking might have been more pleasurable to the animals and had a stronger relaxing effect than "playback." The results regarding the effects of the degree of standardization of the stimulus on the variability of the data were inconclusive. We thus conclude that the use of recorded auditory stimuli to promote positive affective states during human-animal interactions in experimental settings is possible, but not necessarily preferable.

Short- and long-term effects of rearing dairy calves with contact to their mother on their reactions towards humans.

In this Research Communication we address the hypotheses that reduced contact with humans during the first week of life would impair the relationship of dairy calves reared in dam-calf-contact systems to humans in comparison with artificially reared animals, but that this difference would vanish over time. Artificially reared calves (Artificial) that had been separated from their mother within 12 h after birth were bottle-fed with colostrum for 5 d and thereafter sucked milk from an automatic milk feeder. Animals reared with dam-calf contact (Dam-contact) were kept in the calving pen with their dam for 5 d, and then had permanent access to the cow barn and thus to their dam. Calves were weaned at an age of 12 weeks and kept in young stock groups mixed of both treatments until integration into the cow herd. We tested the animals' relationship with humans by assessing the animals' responses towards an unfamiliar person in an avoidance distance (AD) test in the home environment at 4 weeks of age, at 15 months and at 33 months. In calves, we additionally measured AD in a novel arena after a stationary person test. Artificial animals had lower AD, i.e. showed lower level of fear, than Dam-contact calves. However, the AD in Dam-contact calves decreased with increasing number of days they experienced assistance for suckling. Further, there was no significant difference in later ages. In conclusion, gentle human contact in combination with feeding during the first 5 d of life improved calves' relationship to humans leading to differences between the two treatments as well as within the Dam-contact calves. Potential effects under different conditions regarding quantity and quality of human-animal interactions need further research.

Play and social behaviour of calves with or without access to their dam and other cows.

The aim of the study reported in this Research Communication was to compare play behaviour and social interactions of dairy calves either separated from their mother and reared in a calf group (Artificial) or with access to their mother and the cow herd (cow-calf contact: Contact). Contact calves had access to a calf area and also to the cow barn where they could suckle their dam. Artificial calves were fed whole milk up to 16 kg per day via an automatic milk feeder and were only kept in the calf area. We observed the animals on 3 d during the first three months of life. Contact calves showed solitary play, consisting predominantly of locomotor play, for longer than Artificial calves and mainly in the cow barn. This indicates higher welfare in Contact calves. In addition, Artificial calves hardly experienced any agonistic interaction, while Contact calves both initiated and received agonistic interactions, which might contribute to the development of higher social competence.

Methodology for studying human attitudes and behaviour to cow-calf contact systems.

This position paper describes a common stand on methodology of human attitudes and behaviour that is suitable to use in studies regarding cow-calf contact (CCC) in dairy production, in order to create a common knowledge base and foundation for future recommendations of CCC systems. We describe how different quantitative and qualitative methods can be used to study human attitudes to CCC as well as farmer or consumer behaviour. We aim to contribute to a better understanding of the available methods, and hope that this paper can be used as a guideline for future studies in this area.

Methodological terminology and definitions for research and discussion of cow-calf contact systems.

Due to increasing public concern regarding separation of the dairy cow and calf within the first days after birth, alternative systems, where cows and calves stay in contact for an extended period, are receiving increasing interest from a broad array of researchers and other stakeholders. With more research in the area, there is a risk of inconsistencies emerging in the use of terminology. To create a better consensus in further discussions, the aim of this Research Reflection is to provide definitions and propose a common terminology for cow-calf contact in dairy production. We also suggest definitions for various systems allowing cow-calf contact and describe the distinct phases of cow-calf contact systems.

Methodology for experimental and observational animal studies in cow-calf contact systems.

In this Research Reflection we describe a common standpoint on suitable methodology for controlled and observational studies in cow-calf contact systems in dairy production. Different methods to assess behaviour, health and production in cow-calf contact systems are outlined. Knowledge and experience from researchers working in this field supplement scientific literature whenever relevant. Specific methods including study design, early behaviour of cow and calf, social behaviour relevant to cow-calf contact systems, human-animal relationships and aspects related to management (milking, weaning and separation, health) are reviewed, and recommendations formed. We expect that this paper can contribute to a better understanding of the complexity of cow-calf contact systems and help to advance research in this area of dairy production.

Nociceptive Threshold of Calves and Goat Kids Undergoing Injection of Clove Oil or Isoeugenol for Disbudding.

In this preliminary study, we compared changes in mechanical nociceptive thresholds (MNT) of calves and goat kids injected with clove oil or isoeugenol under the horn bud as a potential, more welfare-friendly alternative to hot-iron disbudding. Twenty male calves and goat kids were randomly allocated to clove oil (n = 10 per species) or isoeugenol (n = 10 per species) injection under the horn buds. MNT was measured via a pressure algometer in calves and kids at several locations around the horn buds at several time points before and up to 24 h after injection. In kids, von Frey filaments were used additionally at the same time points. In calves, linear mixed models revealed an effect on MNT of time point (p = 0.010) and side (p = 0.007), but not of injection (p = 0.298), nor of the interaction 'injection*time point', MNT waslowest 9 h post-injection. In goats, there was an effect of injection depending on time point (interaction injection*time point, p = 0.03) with MNT being lowest 24 h post-injection for clove oil, while MNT was similar to pre-injection in isoeugenol. In both species, variation in the individual response post-injection was very high. Our results suggest that clove oil and isoeugenol induced hypersensitivity, which was higher for clove oil, in goat kids, but they also suggest a transient anaesthetic effect in some animals and locations.

Effects of Different Stroking Styles on Behaviour and Cardiac Parameters in Heifers.

Gentle animal-human interactions, such as stroking, can promote positive emotions and thus welfare in cattle. While previous studies showed that stroking at the ventral neck elicited the most positive reactions in cows, intra-specific allogrooming in cattle includes different body regions and is probably guided partly by the receiver. Thus, we compared heifers' (n = 28) reactions to stroking with the experimenter either reactively responding to perceived momentary preferences of the heifers or exclusively stroking the ventral neck. Independently of the stroking style, longer durations of neck stretching and contact occurred during stroking, supporting our hypothesis of a positive perception of stroking. We did not confirm the predicted decrease in heart rate and increase in heart rate variability, but instead found a slightly increased mean heart rate during stroking. The different stroking styles elicited differences in the heifers' ear positions: "reactive" stroking led to longer durations of low ear positions during stroking, while during "ventral neck" stroking, the duration of back up increased. However, no other behaviours differed significantly between different stroking styles, indicating that the exact manner of stroking applied in our treatments seemed to be less important in the promotion of positive affective states in cattle through gentle human-animal interactions.

Salivary oxytocin in pigs, cattle, and goats during positive human-animal interactions.

Research on oxytocin (OT) has provided crucial insight into the neuroendocrine mechanisms of social behavior and human-animal interactions. Nonetheless, this peptide hormone remains challenging to sample, usually requiring invasive measures in the brain or blood, which limits its use. We conducted an analytical validation of the measurement of OT in the saliva of three common farm animal species: pigs, cattle and goats. We then investigated the biological significance of salivary OT concentrations by using a positive human-animal interaction paradigm. The paradigm varied in terms of human familiarity and included a control condition where the animal was alone in the testing pen. A total of 18 pigs, 18 cattle and 9 goats were subjected to each condition using a within-individual counterbalanced design. Animals were free to choose to interact with the human. Saliva samples were collected immediately before and after the 10-min test, and behavior during the test was analyzed. Oxytocin could be reliably measured by ELISA in the saliva of goats (without extraction), and in the saliva of pigs and cattle after extraction. Salivary OT concentration did not change consistently according to testing condition. However, salivary OT concentration was significantly associated with some behaviors, with different relationships depending on the species. In conclusion, OT can be reliably measured in the saliva of goats and, after extraction, in the saliva of pigs and cattle. The relationships of OT with human-animal interactions and animal behavior were complex and to some extent species-dependent, possibly reflecting the perception of the interaction by the animal.

The Effects of Play Behavior, Feeding, and Time of Day on Salivary Concentrations of sIgA in Calves.

The focus of animal welfare science has shifted over the last decades from efforts to avoid negative states to ways of allowing animals the experience of positive emotions. They may influence physiological processes in farmed animals, potentially providing health benefits; in addition, the physiological changes might be used as indicators of emotional states. We investigated calves' salivary secretory immunoglobulin A (sIgA) concentrations with regard to a possible circadian rhythm and two situations that elicit positive emotions. Ten saliva samples of 14 calves were taken on two consecutive days; within the course of a day we observed a significant decline in salivary sIgA concentrations at 14:00 h. Further, we probed the animals before and after milk feeding and, contrarily to our prediction, detected lower sIgA concentrations 5 min after feeding than 15 min before. A probable explanation might be an increase in salivary flow rate caused by milk ingestion. We also took samples before and after we stimulated play behavior in calves. There was no significant difference in sIgA concentrations between samples taken before and after play. Although there was a significant correlation between the change in sIgA concentrations and the amount of play behavior shown, the correlation depended on an unexpected decrease of sIgA in animals that played little, and thus, does not support our hypothesis. In general, the data showed a large variability that might arise from different factors that are difficult to standardize in animals. Thus, the use of salivary sIgA concentrations as a marker of positive emotions in calves is not supported conclusively by the present data.