Odor exploration behavior of the domestic pig (Sus scrofa) as indicator of enriching properties of odors.

Introduction and aim: Although the sense of smell in pigs is widely recognized as being highly developed, surprisingly little is known about their sensory ability. This study aimed to (a) identify which non-social odors pigs were able to detect and distinguish between, (b) investigate the types of behavior expressed when exploring odors and, (c) compare pigs' responses to the different odors to evaluate their interest in the odors. Methods: Growing pigs (N = 192) of crossbred commercial breeds were enrolled in the experiment (32-110 days of age, weighing 64.9 ± 10.1kg). Littermate pairs of opposite sex were tested in test pens with two odor insertion points in the pen wall, 55 cm apart. All pigs were habituated to the test pens and experimenters. Twelve odors were tested (eight essential oils and four synthetic perfumes) in groups of three odors, with each pig pair tested once with one set of three odors (all possible orders of the three odors were tested on 24 pairs in total), always against a non-odor control (demineralized water). In a test, each of the three odors were presented during three trials in a row (a total of 9 trials per test; trial duration: 1 min; inter-trial breaks: 2 min; total test duration: 25 min). Response variables included: duration of sniffing, feeding-related behavior (licking, biting and rooting), agonistic behavior (biting, displacement and pushing) and no approach of the odor or control, recorded throughout each 1-min odor presentation. Results: All pigs sniffed an odor less when repeatedly presented (LMM: all odors P < 0.05), and significantly longer at the subsequent presentation of a new odor [LMM (3rd vs. 1st presentations): P < 0.001]. Specific odor and odor type (essential oil vs. synthetic perfume) had no significant effect on sniffing duration. Overall, feeding-related behavior and agonistic behavior were expressed significantly more when pigs explored the odor compared with the control insertion point (Paired t-tests: P < 0.001), and specific odor only affected the expression of feeding-related behavior. Conclusion: Collectively, pigs express sniffing, agonistic, and feeding-related behavior when exploring odors, which suggests that pigs perceive odors of non-social origin as a resource. Odors may thus constitute relevant enrichment material for pigs.

Erratum: As a mature scientific discipline animal welfare must be subject to debate and opinion - ERRATUM.

[This corrects the article DOI: 10.1017/awf.2023.89.].

Browse or browsing: Investigating goat preferences for feeding posture, feeding height and feed type.

Goats naturally browse different forages in various postures; this differs from typical farm practice, thus there are opportunities to improve goat welfare by understanding what and how they like to eat. We investigated if feeding preference was related to posture, feeder height relative to the ground, and type of feed. Sixteen adult, Saanen cross females participated in two experiments comparing a floor-level feeder (grazing posture; farm standard), with an elevated feeder (browsing posture; Exp1) and a platform-level feeder (raised, grazing posture; Exp2), when two forages (leaves, grass) were offered. Measurements included feed intake (g of DM/feeder), feeder switching frequency, first feeder visited, latency to visit first feeder and exploration and non-feeding activity time. Effects of posture (Exp1), height (Exp2) and feed type were analyzed. Type of feed affected preference for feeding posture and height. All goats consumed leaves over grass (Exp1: POP: 188 ± 6.52 g, GRA: 20.3 ± 7.19 g; Exp2: POP: 191 ± 6.15 g, GRA: 0.231 ± 6.91 g; P < 0.001), and the feeder containing leaves was often visited first (Exp 1: GRA/POP: 94% of visits, P < 0.001, POP/GRA: 53%, P = 0.724; Exp 2: GRA/POP: 91%, P < 0.001; POP/GRA: 69%, P = 0.041). When goats received only leaves, they consumed more from the floor-level (162 ± 22.2 g) vs. elevated level (102 ± 21.9 g) feeder (P = 0.039). When goats received only grass, there was no posture or height preference; however, they changed feeders more frequently (at least 4x (Exp1) and 2x (Exp2) more than other combinations; P > 0.01). Feed intake was negatively affected by exploring time (Exp1 only: r = -0.541; P < 0.001) and performing non-feeding activities (Exp1: r = -0.698; P < 0.001; Exp2: r = -0.673; P < 0.001). We did not identify a preference for elevated feeding posture; however, we suggest that our short test (compared to previous work) encouraged goats to make choices based on line-of-sight and also that the elevated feeder design (replicated from previous work) made leaf access harder. Nonetheless, we highlight that some goats actively used the elevated feeder; this coupled with the clear preference for leaves over grass, suggests that offering feed type and presentation diversity would allow individuals to express their natural feeding behavior more fully.

Seven steps to enhance Open Science practices in animal science.

The Open Science movement aims at ensuring accessibility, reproducibility, and transparency of research. The adoption of Open Science practices in animal science, however, is still at an early stage. To move ahead as a field, we here provide seven practical steps to embrace Open Science in animal science. We hope that this paper contributes to the shift in research practices of animal scientists towards open, reproducible, and transparent science, enabling the field to gain additional public trust and deal with future challenges to guarantee reliable research. Although the paper targets primarily animal science researchers, the steps discussed here are also applicable to other research domains.

Stage 1 Registered Report: Refinement of tickling protocols to improve positive animal welfare in laboratory rats.

Rat tickling is a heterospecific interaction for experimenters to mimic the interactions of rat play, where they produce 50 kHz ultrasonic vocalisations (USV), symptoms of positive affect; tickling can improve laboratory rat welfare. The standard rat tickling protocol involves gently pinning the rat in a supine position. However, individual response to this protocol varies. This suggests there is a risk that some rats may perceive tickling as only a neutral experience, while others as a positive one, depending on how tickling is performed. Based on our research experiences of the standard tickling protocol we have developed a playful handling (PH) protocol, with reduced emphasis on pinning, intended to mimic more closely the dynamic nature of play. We will test whether our PH protocol gives rise to more uniform increases in positive affect across individuals relative to protocols involving pinning. We will compare the response of juvenile male and female Wistar rats as: Control (hand remains still against the side of the test arena), P0 (PH with no pinning), P1 (PH with one pin), P4 (PH with four pins). P1 and P4 consist of a background of PH, with treatments involving administration of an increasing dosage of pinning per PH session. We hypothesise that rats exposed to handling protocols that maximise playful interactions (where pinning number per session decreases) will show an overall increase in total 50 kHz USV as an indicator of positive affect, with less variability. We will explore whether behavioural and physiological changes associated with alterations in PH experience are less variable. We propose that maximising the numbers of rats experiencing tickling as a positive experience will reduce the variation in response variables affected by tickling and increase the repeatability of research where tickling is applied either as a social enrichment or as a treatment.

Impacts of the COVID-19 pandemic on animal behaviour and welfare researchers.

The COVID-19 pandemic has resulted in many changes in the way research is conducted. Some specific groups (e.g. women) and activities (e.g. teaching) may have been disproportionally affected. Our aim was to assess the impact of the COVID-19 pandemic on animal behaviour and welfare researchers' work experience and productivity, focussing on gender, care role, career stage and teaching load. An online survey asked researchers about childcare, research and teaching load and associated changes due to the pandemic, among others, and included the Perceived Stress Scale (PSS) and the Inventory of Socially Supportive Behaviours (ISSB). From June-July 2020, 117 completed responses were received from 28 countries. Time available for writing papers and grants either increased (36 %), decreased (31 %) or these tasks were halted completely (12 %). Perceived productivity was significantly lower for caregivers (P < 0.001) and for men as compared to women (P < 0.001); and low productivity was associated with more stress (higher PSS: P < 0.001). Respondents' experience of the pandemic related to the PSS (b = -0.03 ± 0.02; P = 0.03) and to self-assessed personality traits (P = 0.01). The average PSS of 21 ± 6.5 was greater than the reference value of 15, and was higher when respondents had low job security (P < 0.001) and when they more strongly characterised themselves as perfectionists, hard-working, empathetic and worried (P = 0.02). Respondents who had an intense care role received most social support (P = 0.04). Teaching load increased for 25 % of the respondents but did not significantly relate to any of the response variables. Overall, caregivers and early career researchers faced the most difficulties, and personality traits had a major impact on the ability to cope with the changes caused by the pandemic.

Sensory Abilities of Horses and Their Importance for Equitation Science.

Vision, hearing, olfaction, taste, and touch comprise the sensory modalities of most vertebrates. With these senses, the animal receives information about its environment. How this information is organized, interpreted, and experienced is known as perception. The study of the sensory abilities of animals and their implications for behavior is central not only to ethology but also to animal welfare. Sensory ability, perception, and behavior are closely linked. Horses and humans share the five most common sensory modalities, however, their ranges and capacities differ, so that horses are unlikely to perceive their surroundings in a similar manner to humans. Understanding equine perceptual abilities and their differences is important when horses and human interact, as these abilities are pivotal for the response of the horse to any changes in its surroundings. This review aims to provide an overview of the current knowledge on the sensory abilities of horses. The information is discussed within an evolutionary context and also includes a practical perspective, outlining potential ways to mitigate risks of injuries and enhance positive horse-human interactions. The equine sensory apparatus includes panoramic visual capacities with acuities similar to those of red-green color-blind humans as well as aural abilities that, in some respects exceed human hearing and a highly developed sense of smell, all of which influence how horses react in various situations. Equine sensitivity to touch has been studied surprisingly sparingly despite tactile stimulation being the major interface of horse training. We discuss the potential use of sensory enrichment/positive sensory stimulation to improve the welfare of horses in various situations e.g. using odors, touch or sound to enrich the environment or to appease horses. In addition, equine perception is affected by factors such as breed, individuality, age, and in some cases even color, emphasizing that different horses may need different types of management. Understanding the sensory abilities of horses is central to the emerging discipline of equitation science, which comprises the gamut of horse-human interactions. Therefore, sensory abilities continue to warrant scientific focus, with more research to enable us to understand different horses and their various needs.

Odour conditioning of positive affective states: Rats can learn to associate an odour with being tickled.

Most associative learning tests in rodents use negative stimuli, such as electric shocks. We investigated if young rats can learn to associate the presence of an odour with the experience of being tickled (i.e. using an experimenter's hand to mimic rough-and-tumble play), shown to elicit 50 kHz ultrasonic vocalisations (USVs), which are indicative of positive affect. Male, pair-housed Wistar rats (N = 24) were all exposed to two neutral odours (A and B) presented in a perforated container on alternate days in a test arena. Following 60s of exposure, the rats were either tickled on days when odour A (n = 8) or odour B (n = 8) was present, or never tickled (n = 8). When tickled, rats produced significantly more 50 kHz USVs compared to the days when not being tickled, and compared to control rats. The level of anticipatory 50 kHz USVs in the 60s prior to tickling did not differ significantly between the tickled and control rats. As a retrieval test following the odour conditioning, rats were exposed successively in the same arena to three odours: an unknown neutral odour, extract of fox faeces, and either odours A or B. Compared to controls, 50 kHz USVs of tickled rats increased when exposed to the odour they had previously experienced when tickled, indicating that these rats had learned to associate the odour with the positive experience of being tickled. In a test with free access for 5 min to both arms of a T-maze, each containing one of the odours, rats tickled with odour A spent more time in the arm with this odour. This work is the first to test in a fully balanced design whether rats can learn to associate an odour with tickling, and indicates that positive odour conditioning has potential to be used as an alternative to negative conditioning tests.

Effects of choice feeding and lower ambient temperature on feed intake, growth, foot health, and panting of fast- and slow-growing broiler strains.

Effects of lowering ambient temperature in the second half of the growth period were studied in 2 broiler strains with different genetic potential for growth (Ross from 1972 and 2004) when given ad libitum access or pre-mixed increasing proportion of whole wheat in their diet. A total of 48 groups of day-old broilers (n = 64) from the 2 strains were housed as hatched in 48 pens in 4 houses and slaughtered at 6 wk of age. Half of the groups had access to pelleted feed pre-mixed with increasing proportion of whole wheat, the other half had access to pelleted feed and whole wheat in separate troughs. Ambient temperature from 28 d of age was either 21 or 15°C. The fast-growing strain had greater (P < 0.001) live weight throughout (mean slaughter weights 2.6 (±0.014) kg and 1.0 (±0.006) kg). When given a choice, the fast-growing strain consumed a lower (P ≤ 0.004) proportion of whole wheat in their diet in weeks 5 and 6 (20 and 26 (±0.9) % whole wheat), and the slow-growing strain chose a higher (P < 0.001) wheat proportion (45 and 56 (±1.7) %) than the pre-mixed inclusion of 25 and 30%, respectively. The slow-growing strain grew slightly faster in the colder ambient temperature, and slower when given a free choice of whole wheat. Both strains drank more water per feed consumed in the warmer ambient temperature treatment. Foot pad dermatitis, only seen in the fast-growing birds, had a higher occurrence in the colder and in the choice feeding treatments. Gait scoring on day 34 showed perfect gait in more than 85% of the slow-growing and less than 1% of the fast-growing birds. Activity was higher and panting levels were lower in the slow-growing strain (P < 0.001) toward the end of the growth period, with no effect of the temperature treatment on panting. Slow-growing strains increase their energy intake, and fast-growing strains increase their protein intake when given a choice.

Sexual responses of male rats to odours from female rats in oestrus are not affected by female germ-free status.

Rats detect and use odorant molecules as a source of information about their environment. Some of these molecules come from conspecifics, and many arise as by-products from microbial activity. Thus, compared to conventionally housed rats, germ-free rats are raised in an environment with fewer odorants, but this reduction is rarely quantified. Using gas chromatography-mass spectrometry, we found that germ-free rat faeces samples contained half as many volatile molecules than conventional rat faeces (52 vs 109 (±2.4) molecules; P < 0.001) and overall these were only 12% as abundant. We then investigated if odours from female germ-free rats in oestrus would have pro-erectile effects in conventional male rats. For this aim, conventionally housed Brown Norway (BN) rats (n = 16) with sexual experience with either Fischer or BN females, were exposed to four different odour types: faeces from germ-free Fischer rat in oestrus, faeces from conventional rats in oestrus and di-oestrus (either from Fischer or BN), and a control (either 1-hexanol or male rat faeces). The number of penile erections per test as well as the duration of freezing behaviour was significantly higher with the oestrous odours (germ-free and conventional) compared to the control, with intermediate responses to the di-oestrous faeces. The findings indicate that, despite a significantly reduced composition in terms of volatiles compared to conventionally housed rats, the faeces of germ-free rats contain sufficient odorants to evoke sexual responses in conventional male rats. Oestrous odours of rats thus appear not to be of microbial origin.

Prepartum Maternal Behavior of Domesticated Cattle: A Comparison with Managed, Feral, and Wild Ungulates.

The event of giving birth is an essential part of animal production. In dairy cattle production, there are substantial economical and welfare-related challenges arising around the time of parturition, and hence increased focus on efficient management of the calving cow. Drawing on the research literature on prepartum maternal behavior, this review compares cattle to other members of the ungulate clade with the aim of understanding the biological basis of bovine prepartum behavior with main emphasis on dairy cows. Ultimately, this knowledge may be used in future development of housing systems and recommendations for the management of calving cows. Maternal prepartum behavior varies among species, but the final goal of ungulate mothers is the same: ensuring a calm parturition and optimal environment for the onset of postpartum maternal behavior by locating an appropriate birth site, with low risk of predators, disturbances and mistaken identity of offspring. Features of chosen birth sites vary among species and depend largely on the environment, as ungulate females display a considerable ability to adapt to their surroundings. However, within commercial housing conditions in dairy production, the animals' ability to adapt behaviorally appears to be challenged. Confinement alongside high stocking densities leave little room to express birth-site selection behavior, posing a high risk of agonistic social behavior, disturbances, and mismothering, as well as exposure to olfactory cues influencing both prepartum and postpartum maternal behavior. Dairy cows are thus exposed to several factors in a commercial calving environment, which may thwart their maternal motivations and influence their behavior. In addition, prepartum cattle may be more affected by olfactory cues than other ungulate species (e.g., sheep) because they are attracted to birth fluids already before calving. Hence, providing dairy cows with an environment where they can perform the maternal behavior they are motivated for, may aid a calm and secure calving and provide optimal surroundings for postpartum maternal behavior. Future research should focus on designing motivation-based housing systems allowing freedom to express prepartum maternal behavior and investigate in more detail the effects of the environment on the welfare of calving cows and their offspring.

Where is the TMT? GC-MS analyses of fox feces and behavioral responses of rats to fear-inducing odors.

TMT (2,5-dihydro-2,4,5-trimethylthiazoline) is known as a component of fox feces inducing fear in rodents. However, no recent chemical analyses of fox feces are available, and few studies make direct comparisons between TMT and fox feces. Fox feces from 3 individuals were used to prepare 24 samples to be analyzed for the presence of TMT using gas chromatography-mass spectrometry (GC-MS). When TMT was added in low amounts (50-2000 nmol/g), TMT was detected in 10 out of 11 samples. When no TMT was added, TMT was detected in only 1 out of 13 samples. In a second experiment, we tested the behavioral response of male Brown Norway (BN) and Wistar rats to either fox feces, a low amount of TMT (0.6 nmol) or 1-hexanol. TMT induced freezing in the rats, but fox feces induced significantly more freezing episodes and longer total duration of freezing in both rat strains. In experiment 3, male BN rats were exposed over several days to fox feces, rat feces, 1-hexanol, cadaverine, 2-phenylethylamine, and TMT, one odor at a time. Fox feces induced significantly more freezing episodes of a longer total duration than any of the other odors, with rat feces and 1-hexanol giving rise to the lowest amount of freezing. This finding, together with our inability to verify the presence of TMT in fox feces, indicates that the concentration of TMT in our fox feces samples was below 50 nmol/g. It may also be that other compounds in fox feces play a role in its fear-inducing properties.

Lameness Affects Cow Feeding But Not Rumination Behavior as Characterized from Sensor Data.

Using automatic sensor data, this is the first study to characterize individual cow feeding and rumination behavior simultaneously as affected by lameness. A group of mixed-parity, lactating Holstein cows were loose-housed with free access to 24 cubicles and 12 automatic feed stations. Cows were milked three times/day. Fresh feed was delivered once daily. During 24 days with effectively 22 days of data, 13,908 feed station visits and 7,697 rumination events obtained from neck-mounted accelerometers on 16 cows were analyzed. During the same period, cows were locomotion scored on four occasions and categorized as lame (n = 9) or not lame (n = 7) throughout the study. Rumination time, number of rumination events, feeding time, feeding frequency, feeding rate, feed intake, and milk yield were calculated per day, and coefficients of variation were used to estimate variation between and within cows. Based on daily sums, using each characteristic as response, the effects of lameness and stage of lactation were tested in a mixed model. With rumination time as response, each of the four feeding characteristics, milk yield, and lameness were tested in a second mixed model. On a visit basis, effects of feeding duration, lameness, and milk yield on feed intake were tested in a third mixed model. Overall, intra-individual variation was <15% and inter-individual variation was up to 50%. Lameness introduced more inter-individual variation in feeding characteristics (26-50%) compared to non-lame cows (17-29%). Lameness decreased daily feeding time and daily feeding frequency, but increased daily feeding rate. Interestingly, lameness did not affect daily rumination behaviors, fresh matter intake, or milk yield. On a visit basis, a high feeding rate was associated with a higher feed intake, a relationship that was exacerbated in the lame cows. In conclusion, cows can be characterized in particular by their feeding behavior, and lame cows differ from their non-lame pen-mates in terms of fewer feed station visits, faster eating, less time spent feeding, and more variable feeding behavior. Further, daily rumination time was slightly negatively associated with feeding rate, a relationship which calls for more research to quantify rumination efficiency relative to feeding rate.

Behavioral responses to odors from other species: introducing a complementary model of allelochemics involving vertebrates.

It has long been known that the behavior of an animal can be affected by odors from another species. Such interspecific effects of odorous compounds (allelochemics) are usually characterized according to who benefits (emitter, receiver, or both) and the odors categorized accordingly (allomones, kairomones, and synomones, respectively), which has its origin in the definition of pheromones, i.e., intraspecific communication via volatile compounds. When considering vertebrates, however, interspecific odor-based effects exist which do not fit well in this paradigm. Three aspects in particular do not encompass all interspecific semiochemical effects: one relates to the innateness of the behavioral response, another to the origin of the odor, and the third to the intent of the message. In this review we focus on vertebrates, and present examples of behavioral responses of animals to odors from other species with specific reference to these three aspects. Searching for a more useful classification of allelochemical effects we examine the relationship between the valence of odors (attractive through to aversive), and the relative contributions of learned and unconditioned (innate) behavioral responses to odors from other species. We propose that these two factors (odor valence and learning) may offer an alternative way to describe the nature of interspecific olfactory effects involving vertebrates compared to the current focus on who benefits.

Using scalar products to refine the interpretative value of an orientation choice test.

Choice tests, which are often used to examine animal preferences, can be difficult to interpret when no clear choice has been made or when using very young animals which exclude test repetition. We present a new method to evaluate the behavior in a choice test based on the orientation of the animal and illustrate its use when facing those conditions. Using rat pups in an open field maze with a choice of odors, we obtained x,y coordinates of 2 markers (head and body center) using a video-tracking freeware. Two vectors were calculated: an animal orientation vector (body to head) and a perfect orientation vector (body to odor source). The angle between the 2 vectors in each frame was converted into a scalar product ranging from 1 (pup oriented directly towards the odor source) to -1 (facing the opposite direction). A mean scalar product was calculated for each odor source, with the difference between the 2 mean scalar products indicating degree of preference for an odor. The information provided by the mean scalar product difference (MSPD) could not be obtained from other measures, such as binary choice, velocity, or distance moved. The MSPD provides a single, noncategorical value for each animal to describe degree of preference in a choice test. This variable was more effective in differentiating animals, thus allowing a reduction in the number of animals or tests necessary to reach significance.

Behavioural response of sexually naïve and experienced male rats to the smell of 6-methyl-5-hepten-2-one and female rat faeces.

Sexually experienced male rats display penile erections when exposed to faeces from mammalian females in oestrus (Rampin et al., Behav Brain Res, 172:169, 2006), suggesting that specific odours indicate female receptiveness across species. However, it is unknown to what extent the sexual response observed results from an odorous conditioning acquired during sexual experience. We tested the behavioural response of male Brown Norway rats both when sexually naïve and experienced to four odours, including oestrous rat faeces and 6-methyl-5-hepten-2-one (methylheptenone; a molecule found in higher concentrations during oestrus in female rats, foxes and horses). Odour had a significant effect on the sexual response of the naïve rats, with oestrus faeces provoking significantly more erections than herb odour, and with methylheptenone and di-oestrus faeces being intermediate. This indicates that sexually naïve male rats have an unconditioned ability to detect oestrous mediated via odour. After gaining sexual experience, the response to methylheptenone, di- and oestrus faeces was significantly higher than that observed with herb odour. These results strongly suggest that methylheptenone is part of the odorous bouquet of oestrus and contributes to the olfactory determination of female receptiveness.

A mixture of odorant molecules potentially indicating oestrus in mammals elicits penile erections in male rats.

A common set of odorous molecules may indicate female receptiveness across species, as male rats display sexual arousal when exposed to the odour of oestrous faeces from rats, vixens and mares. More than 900 different compounds were identified by GC-MS analyses performed on faeces samples from di-oestrous and oestrous females and from males of the three species. Five carboxylic acids were found in lower concentrations in faeces from all oestrous females. We subjected 12 sexually trained male rats to a 30 min exposure to different dilutions of a mixture of these five molecules in the same proportions as found in female oestrous faeces. The behavioural responses of the rats were compared to those displayed when exposed to water (negative control) and faeces from oestrous female rats (positive control). Frequency of penile erections were found to be significantly dependent on mixture dilution, with two intermediate dilutions eliciting frequencies of penile erections that did not differ from those obtained during exposure to oestrous female rat faeces. Higher and lower dilutions did not elicit more penile erections than observed with water. These results support our hypothesis that a small set of odorous molecules may indicate sexual receptiveness in mammalian females.