Lameness in Pregnant Sows Alters Placental Stress Response.

Pregnant sows from commercial pig farms may experience painful states, such as lameness, an essential indicator in assessing sow welfare. We investigated the effect of lameness during the last third of pregnancy on reproductive performance and placental glucocorticoid concentrations in sows. Periodic locomotion assessments were carried out on two commercial pig farms using a validated 0-5 scoring system (from 0: normal locomotion; to 5: a downer animal). Sows from both farms (N = 511) were grouped based on their average locomotion scores. On Farm 1, 30 sows were selected and grouped as either Not Lame (NL = 16; X¯ = 0-1) or Lame (L = 14; X¯ > 1). On Farm 2, 39 sows were selected and grouped as either Not Lame (G1 = 12; X¯ = 0-1), Moderately Lame (G2 = 13; X¯ = 1.1-2), or Severely Lame (G3 = 14; X¯ ≥ 2.1). Reproductive data (gestation length, litter weight, average piglet weight, litter size, and the number of piglets born alive/mummified/stillborn) were recorded on both farms. Moreover, on Farm 2, piglet intrauterine growth restriction score and the number of piglets dead during the first week were also recorded, and placenta samples were collected to determine their cortisol/cortisone concentrations. A linear mixed model was used to analyze the data. The proportion of lameness in pregnant sows (N = 511) was >40%, and the gestation length tended to decrease with the presence of lameness (p < 0.1) in both farms. G2 sows had a higher placental cortisol/cortisone ratio than G1 and G3 sows (p < 0.01). In conclusion, lameness was high in the sows assessed, which may decrease sow gestation length and reduce placental efficiency in protecting the offspring from the sows' stress response.

Non-invasive estimation of in vivo optical properties and hemodynamic parameters of domestic animals: a preliminary study on horses, dogs, and sheep.

Biosensors applied in veterinary medicine serve as a noninvasive method to determine the health status of animals and, indirectly, their level of welfare. Near infrared spectroscopy (NIRS) has been suggested as a technology with this application. This study presents preliminary in vivo time domain NIRS measurements of optical properties (absorption coefficient, reduced scattering coefficient, and differential pathlength factor) and hemodynamic parameters (concentration of oxygenated hemoglobin, deoxygenated hemoglobin, total hemoglobin, and tissue oxygen saturation) of tissue domestic animals, specifically of skeletal muscle (4 dogs and 6 horses) and head (4 dogs and 19 sheep). The results suggest that TD NIRS in vivo measurements on domestic animals are feasible, and reveal significant variations in the optical and hemodynamic properties among tissue types and species. In horses the different optical and hemodynamic properties of the measured muscles can be attributed to the presence of a thicker adipose layer over the muscle in the Longissimus Dorsi and in the Gluteus Superficialis as compared to the Triceps Brachii. In dogs the absorption coefficient is higher in the head (temporalis musculature) than in skeletal muscles. The smaller absorption coefficient for the head of the sheep as compared to the head of dogs may suggest that in sheep we are indeed reaching the brain cortex while in dog light penetration can be hindered by the strongly absorbing muscle covering the cranium.

Events with Different Emotional Valence Affect the Eye's Lacrimal Caruncle Temperature Changes in Sheep.

Infrared thermography (IRT) has been recently applied to measure lacrimal caruncle temperature non-invasively since this region is related to the sympathetic response, and it seems a promising technique that is able to infer negative emotions in sheep (e.g., fear). However, the scientific literature so far is limited in understanding whether a caruncle's temperature changes also in response to positive emotional states in sheep. Through classical conditioning, we aimed to assess how a positive or a negative event affects the physiological (lacrimal caruncle temperature measured with IRT and cortisol levels) and behavioral responses of sheep (ear position). Fourteen ewes from the same flock were randomly assigned to two treatment groups: positive (n = 7) and negative (n = 7). Each group was then trained through classical conditioning to associate a neutral auditory (ring bell) stimulus to an oncoming event: for the positive group, the presence of a food reward (maize grains), while for the negative one, the opening of an umbrella. After three weeks of training, before (at rest) and after (post-treatment), lacrimal caruncle temperature was non-invasively measured via IRT, and saliva samples were gently collected to measure cortisol levels. During treatment, sheep behavior was videorecorded and then analyzed using a focal animal sampling technique. At rest, the eye's lacrimal caruncle temperature was similar in both groups, while post-treatment, a significant increase was shown only in the negative group (t-test; p = 0.017). In the anticipation phase, sheep in the positive group kept their ears forward longer compared to those in the negative one (Mann-Whitney; p < 0.014), 8.3 ± 2.1 s and 5.2 ± 4.2 s, respectively. The behavioral response observed reflects a learnt association between a neutral stimulus and events with different emotional valence. Cortisol concentration slightly increased in both groups post-treatment. Our results confirm that IRT is a non-invasive technique that can be useful when applied to assess how positive and negative events may affect the physiological response in sheep.

Assessment of Seasonal Variation in Methane Emissions of Mediterranean Buffaloes Using a Laser Methane Detector.

A direct assessment of the methane (CH4) emission level and its variability factors is needed in each animal species in order to target the best mitigation strategy for the livestock sector. Therefore, the present study aimed to (1) test a laser methane detector (LMD) for the first time in Italian Mediterranean buffaloes (IMB), a non-invasive tool to quantify CH4 emissions; (2) test the effect of season on the emissions; and (3) compare the results measured directly with the ones estimated with the existing equations. CH4 emissions of twenty non-productive IMB, under the same feeding regimen, were monitored for 12 days in summer and winter. Significantly higher THI (74.46 ± 1.88 vs. 49.62 ± 4.87; p < 0.001), lower DMI (2.24 ± 0.04 vs. 2.51 ± 0.03% DMI/kg live weight; p < 0.001) and lower emission intensities (0.61 ± 0.15 vs. 0.75 ± 0.13; p < 0.001) were found during the summer period when compared with winter. LMD was found to be a versatile tool to be used in buffaloes, and it was clear that a summer increase in THI could act as a stressor for the animals, influencing their emissions. In addition, measured emissions were significantly higher than when estimated with the existing equations (p < 0.001), suggesting the need for further research in this area.

A Magnetic Resonance-Relaxometry-Based Technique to Identify Blood Products in Brain Parenchyma: An Experimental Study on a Rabbit Model.

Magnetic resonance relaxometry is a quantitative technique that estimates T1/T2 tissue relaxation times. This has been proven to increase MRI diagnostic accuracy of brain disorders in human medicine. However, literature in the veterinary field is scarce. In this work, a T1 and T2-based relaxometry approach has been developed. The aim is to investigate its performance in characterizing subtle brain lesions obtained with autologous blood injections in rabbits. This study was performed with a low-field scanner, typically present in veterinary clinics. The approach consisted of a semi-automatic hierarchical classification of different regions, selected from a T2 map. The classification was driven according to the relaxometry properties extracted from a set of regions selected by the radiologist to compare the suspected lesion with the healthy parenchyma. Histopathological analyses were performed to estimate the performance of the proposed classifier through receiver operating characteristic curve analyses. The classifier resulted in moderate accuracy in terms of lesion characterization.

Animal Welfare and Parasite Infections in Organic and Conventional Dairy Farms: A Comparative Pilot Study in Central Italy.

The study investigated and compared welfare conditions and gastrointestinal (GI) parasites distribution among organic (ORG) and conventional (CONV) farms in central Italy. Five ORG and five CONV farms were assessed for animal welfare with an adapted version of the AssureWel protocol. Faecal samples collected from the rectum of the animals both in ORG (n = 150) and CONV (n = 150) were analysed using conventional copromiscroscopy. The presence of skin damages in the rear legs was significantly predominant (p < 0.001) in CONV (26.7%) compared with ORG farms (10.0%). No differences were found for lameness, cleanliness, Body Condition Score, hair loss, body lesions and swelling prevalence. Data concerning the productive performances, e.g., total milk, fat and protein yields standardised in mature equivalent (ME) were collected. ME milk yield (ORG: 9656.9 ± 1620.7 kg; CONV: 12,047.2 ± 2635.3) and ME fat yield (ORG: 396.6 ± 66.8; CONV: 450.3 ± 102.8) were significantly lower in ORG farms (p < 0.001). Anthelmintics were used regularly in 4/5 CONV and 0/5 ORG farms. In 2 CONV farms (40%) and 4 ORGs (80%) at least one animal tested positive for GI parasites. No significant differences in parasites prevalence emerged (ORG = 10.7%; CONV = 8%). These data indicate that ORG farming does not influence parasite prevalence and animal welfare status.

Evaluation of Brix Refractometry to Estimate Immunoglobulin G Content in Buffalo Colostrum and Neonatal Calf Serum.

Brix refractometry has been widely demonstrated to be a useful tool for monitoring colostrum management program and passive immunity transfer (PIT) in Bovines, but its suitability has never been verified in Buffalo. Therefore, the objective of this study was to evaluate the utility of a simple and rapid tool such as a digital Brix refractometer to estimate colostrum quality and for evaluating the success of passive transfer of immunoglobulin G (IgG) in Buffalo calves. The optimal cut points levels for Brix Refractometry for distinguishing good- and poor-quality colostrum and for assessing the adequacy of passive immunity transfer in calves were determined. For this aim, 26 first-milking maternal colostrum (MC) were collected from first-calf heifers. Blood samples were obtained from their calves at birth (T0) and 72 hours after (T3). Colostrum and Serum IgG content were determined by indirect enzyme-linked immunosorbent assay (ELISA), whereas total protein (TP, g/dL) and percentage Brix (%Brix) by means of a digital Brix refractometer. The mean colostrum IgG was 64.9 ± 29.3 mg/mL. The mean serum %Brix at T3 was 9.6 ± 0.9 %. The mean serum IgG content at T3 was 11.1 ± 2.0 mg/mL. Pearson's correlation coefficient (rp) was determined between Brix and ELISA measurements: colostrum %Brix showed a significant correlation with serum %Brix (rp = 0.82, p < 0.001); serum %Brix was highly correlated with serum TP (STP, g/dL) (rp = 0.98, p < 0.001) and serum IgG (mg/mL) (rp = 0.85, p < 0.001). A cut point of 18% Brix to estimate samples of MC ≥ 50 mg/mL from first-calf heifers was more appropriate for the buffalo. A cut point of 8.4% Brix resulted in the greatest percentage of calf serum samples being correctly classified. Based on our findings, a digital Brix refractometer could be a useful tool to monitor colostrum quality and to estimate PIT in Buffalo calves.

Maternal and Neonatal Behaviour in Italian Mediterranean Buffaloes.

The aim of this study was to describe the neonatal and maternal behaviour of Italian Mediterranean buffaloes. Thirty primiparous buffaloes were moved into individual pens 12.5 (±2.5) days before calving. Maternal and neonatal behaviours were recorded for 48 h after calving and the analysis was performed in continuous sampling with the software BORIS. Calves' clinical evaluations (temperature, weight, and heart and respiratory rates) were performed at different time intervals and correlated with behavioural data from the dam. Data were analysed with parametric and non-parametric methods after controlling their distribution. The maternal behavioural pattern found highlighted buffaloes' priorities during the post-partum period: firstly, they stand and start grooming to ensure proper care for the calf; it is only after this that they dedicate time to maintenance behaviours (feeding and lying). The dams mainly groomed the calf during the first six hours after calving (average time in the 1-6-h interval: 7.7 ± 2.5 min., F = (2.5, 60.2) = 75.0; p < 0.001) to ensure the formation of the mother-infant bond; thereafter, the behaviour decreased over time. As reported in the literature, inexperienced mothers could sometimes delay the calf's first suckling with aggressive or rejection behaviours. In this regard, 16 buffalo dams showed at least one maternal rejection behaviour, which was found to negatively correlate with calves' daily weight gain (DWG) at 14 (rs = -0.5, p = 0.02) and 21 days (rs = -0.7, p < 0.001). The calves took on average 212.0 ± 110.0 min to suckle, and this behaviour was mainly shown during the first six hours. Overall, suckling behaviour was correlated with standing: (rs = 0.6, p < 0.001) and walking (rs = 0.9, p < 0.001). The calves' live weight and DWG were consistently higher than the values reported in the literature. Our results present a detailed description of maternal and neonatal behaviour in the early post-partum period in Italian Mediterranean buffaloes. We also found that maternal rejection behaviours can negatively influence the calves' growth. Finally, we think that such results can improve the management of buffaloes during the period around parturition.

Analysis of Hindgut Microbiome of Sheep and Effect of Different Husbandry Conditions.

The microbiome is now seen as an important resource to understand animal health and welfare in many species. However, there are few studies aiming at identifying the association between fecal microbiome composition and husbandry conditions in sheep. A wide range of stressors associated with management and housing of animals increases the hypothalamic-pituitary axis activity, with growing evidence that the microbiome composition can be modified. Therefore, the purpose of the present study was to describe the core microbiome in sheep, characterized using 16S rRNA gene sequencing, and to explore whether exposure to stressful husbandry conditions changed sheep hindgut microbiome composition. Sheep (n = 10) were divided in two groups: isolated group (individually separated for 3 h/day) and control group (housed in the home pen for the entire trial period). Sheep core microbiome was dominated by Firmicutes (43.6%), Bacteroidetes (30.38%), Proteobacteria (10.14%), and Verrucomicrobia (7.55%). Comparative results revealed few operational taxonomic units (OTUs) with significantly different relative abundance between groups. Chao1, abundance-based coverage estimator (ACE), and Fisher's alpha indices did not show differences between groups. OTU-based Bray-Curtis distances between groups were not significant (p-value = 0.07). In conclusion, these results describing the core microbiome of sheep do not suggest a strong effect of stressful husbandry conditions on microbial composition.

Reliability of fNIRS for noninvasive monitoring of brain function and emotion in sheep.

The aim of this work was to critically assess if functional near infrared spectroscopy (fNIRS) can be profitably used as a tool for noninvasive recording of brain functions and emotions in sheep. We considered an experimental design including advances in instrumentation (customized wireless multi-distance fNIRS system), more accurate physical modelling (two-layer model for photon diffusion and 3D Monte Carlo simulations), support from neuroanatomical tools (positioning of the fNIRS probe by MRI and DTI data of the very same animals), and rigorous protocols (motor task, startling test) for testing the behavioral response of freely moving sheep. Almost no hemodynamic response was found in the extra-cerebral region in both the motor task and the startling test. In the motor task, as expected we found a canonical hemodynamic response in the cerebral region when sheep were walking. In the startling test, the measured hemodynamic response in the cerebral region was mainly from movement. Overall, these results indicate that with the current setup and probe positioning we are primarily measuring the motor area of the sheep brain, and not probing the too deeply located cortical areas related to processing of emotions.

Venous blood gas parameters in healthy Mediterranean buffalo calves in the first 72 hours of life.

In the first hours after calving the neonate radically modifies its anatomy and physiology to adapt to extrauterine life. These modifications, however, make the calf prone to respiratory and metabolic dysfunction. Thus, stillbirth, defined as the death of the neonate within 48 h after birth, represents a relevant issue for buffalo farming. Of the clinical procedures, the blood gas analysis of venous samples provides useful information regarding the acid-base status of the neonate, allowing the optimal management of potential illness calves. However, the blood gas parameters have never been studied in the buffalo calf in the first 72 h of life. Thus, this study aimed to report blood gas parameters at calving and 1, 12, 24, and 72 h after birth in healthy buffalo neonates born after normal parturition. The study involved 20 healthy buffalo calves, born from normal pregnancy and parturition. The parturition and the first hours after were monitored by closed-circuit television cameras, and within 1 min after calving the buffalo neonate received a clinical evaluation (APGAR, temperature), venous blood sampling for blood gas (pH, partial pressure carbon dioxide - pCO2, partial pressure oxygen - pO2, base excess, bicarbonate concentration, oxygen saturation, total carbon dioxide - tCO2, and lactate), and glycemia analysis. The same evaluations were performed after 1, 12, 24, and 72 h. The present study showed a moderate mixed acidosis at calving, similar to that reported in the bovine neonate, likely due to the high venous blood pCO2 and lactate. This acidemia resolved spontaneously within 12 h as a reduction of pCO2 (P ≤ 0.05). Lactate reduced significantly (P ≤ 0.05) only at 72 h, similarly to the trend reported previously for the bovine calf. Blood lactate concentration was correlated with glycemia at calving and 1 h (r = 0.677, P = 0.014), and with stage 2 duration (r = 0.647, P = 0.006). These findings support a possible mechanism of glucose generation induced by catecholamines coupled to anaerobic metabolism soon after calving. After calving, all the blood gas parameters changed rapidly reaching values comparable with those reported in the literature for older buffalo calves from 72 h after calving. No correlations were found between APGAR score and venous blood gas parameters, likely due to the inclusion criteria that selected only health buffalo calves born after normal parturition. The findings suggested that the venous blood gas analysis could be considered supportive rather than an alternative to the clinical evaluation of the buffalo calf.

Phylogenetic variation in cortical layer II immature neuron reservoir of mammals.

The adult mammalian brain is mainly composed of mature neurons. A limited amount of stem cell-driven neurogenesis persists in postnatal life and is reduced in large-brained species. Another source of immature neurons in adult brains is cortical layer II. These cortical immature neurons (cINs) retain developmentally undifferentiated states in adulthood, though they are generated before birth. Here, the occurrence, distribution and cellular features of cINs were systematically studied in 12 diverse mammalian species spanning from small-lissencephalic to large-gyrencephalic brains. In spite of well-preserved morphological and molecular features, the distribution of cINs was highly heterogeneous, particularly in neocortex. While virtually absent in rodents, they are present in the entire neocortex of many other species and their linear density in cortical layer II generally increased with brain size. These findings suggest an evolutionary developmental mechanism for plasticity that varies among mammalian species, granting a reservoir of young cells for the cerebral cortex.

To acquire new skills or recover after injuries, the mammalian brain relies on plasticity, the ability for the brain to change its architecture and its connections during the lifetime of an animal. Creating new nerve cells is one way to achieve plasticity, but this process is rarer in humans than it is in mammals with smaller brains. In particular, it is absent in the human cortex: this region is enlarged in species with large brains, where it carries out complex tasks such as learning and memory. Producing new cells in the cortex would threaten the stability of the structures that retain long-term memories. Another route to plasticity is to reshape the connections between existing, mature nerve cells. This process takes place in the human brain during childhood and adolescence, as some connections are strengthened and others pruned away. An alternative mechanism relies on keeping some nerve cells in an immature, 'adolescent' state. When needed, these nerve cells emerge from their state of arrested development and 'grow up', connecting with the appropriate brain circuits. This mechanism does not involve producing new nerve cells, and so it would be suitable to maintain plasticity in the cortex. Consistent with this idea, in mice some dormant nerve cells are present in a small, primitive part of the cortex. La Rosa et al. therefore wanted to determine if the location and number of immature cells in the cortex differed between mammals, and if so, whether these differences depended on brain size. The study spanned 12 mammal species, from small-brained species like mice to larger-brained animals including sheep and non-human primates. Microscopy imaging was used to identify immature nerve cells in brain samples, which revealed that the cortex in larger-brained species contained more adolescent cells than its mouse counterpart. The difference was greatest in a region called the neocortex, which has evolved most recently. This area is most pronounced in primates ­ especially humans ­ where it carries out high-level cognitive tasks. These results identify immature nerve cells as a potential mechanism for plasticity in the cortex. La Rosa et al. hope that the work will inspire searches for similar reservoirs of young cells in humans, which could perhaps lead to new treatments for brain disorders like dementia.

Effects of three different diets on the fatty acid profile and sensory properties of fresh Pecorino cheese "Primo Sale".

OBJECTIVE: This study aimed to evaluate and compare the effects of three different diets on the fatty acids (FA) profile and sensory properties of a characteristic Italian fresh cheese: Pecorino "Primo Sale" (PS). METHODS: Fifty-four sheep were divided into three feeding groups: total mixed ration (TMR) enriched with extruded linseed (TL), control diet with TMR without any integration (TC), and pasture (P). During cheese production, six cheeses per experimental group were produced each week, stored for 10 days at 4°C, and then analyzed for chemical composition, FA profile, and sensory properties. RESULTS: Saturated fatty acids (SFA) were significantly higher in PS from group TC (82.11%) than in cheese from other two groups (P 75.48% and TL 66.83%). TL and P groups presented higher values of polyunsaturated fatty acids, 4.35 and 3.65%, respectively, than that of TC group (2.31%). The lowest SFA/unsaturated fatty acid ratio was found in TL and P groups, while the highest was found in the TC group. Vaccenic acid and conjugated linoleic acid (CLA) were higher in group P (p<0.05) than in groups L and TC. Sensory properties of cheese from group P received the highest scores for odor intensity and friability, while control group had a greater chewing consistency. Overall, all cheeses received good scores for acceptability. CONCLUSION: In conclusion, this study showed how the integration of extruded linseed improved the FA profile of fresh pecorino cheese PS preserving its sensory properties. Levels of CLA in the PS group achieved using this approach was not higher than that in a grazing diet. Cheeses from groups P and L contained a higher value of FA, with nutritional implications for humans, when compared with un-supplemented diet. Linseed may be a good feeding strategy when pasture is not available.

Effect of Ewe Diet on Milk and Muscle Fatty Acid Composition of Suckling Lambs of the Protected Geographical Origin Abbacchio Romano.

Consumers increasingly pay more attention to the lipid profile of meat products and consume less meat to reduce cholesterol and heart disease. In Italy, sheep producers are increasingly feeding sheep fresh forage. We investigated whether the supplementation of dam diet with extruded linseed would be an alternative strategy to pasture for improving the intramuscular and subcutaneous FA compositions of their suckling lambs. The ewe diets were enriched with either extruded linseed (L), un-supplemented farm diet (F), or pasture (P). Milk saturated fatty acids (SFA) decreased in P and L compared with F, while the opposite pattern was observed for polyunsaturated FA (PUFA) and conjugated linoleic acids after seven days. The FA composition of lamb meat was similar to that of their dam's milk, showing higher PUFA in P and L compared to F, while SFA was higher in F. Regarding the lamb meat obtained from barn-held ewes, L had lower n-6/n-3 content compared to F, while an intermediate content was found in P. These results indicate a better n-3 FA profile in milk and lamb's meat from pasture and linseed-enriched diets. No changes in lamb performance were observed.

Salivary IgG levels in neonatal calves and its association to serum IgG: an observational pilot study.

The diagnosis of inadequate transfer of colostrum immunoglobulin G (IgG) to calf serum, often known as failure of passive transfer (<10 g/L IgG1 at 24 to 48 h), necessitates blood sampling from the calf and in some instances the presence of a veterinarian. Sampling saliva is both less invasive and easy for the producer. Previous research has shown that quantification of saliva IgG is possible in juvenile and adult cattle. The objectives of this observational pilot study were to investigate whether IgG can be quantified in neonatal calf saliva, if it is correlated to serum IgG concentrations, and if the indirect quantification of saliva IgG is achievable by use of a digital refractometer. Paired blood and saliva samples were collected from 20 healthy dairy calves aged 1 to 3 d. In these samples, IgG was quantified directly with single radial immunodiffusion and indirectly by use of a digital refractometer indicating Brix % (a subsample of n = 12 saliva samples). A strong positive correlation (r = 0.7, P < 0.001) between saliva IgG (mean ± SD; 0.2 ± 0.11 g/L) and serum IgG (32.1 ± 11.94 g/L) was found. Saliva IgG ranged from the lowest detectable value, 0.1 g/L (n = 6 samples) to 0.6 g/L. Saliva Brix (1.2 ± 0.69%) was not significantly correlated to serum IgG (n = 12, r = 0.43, P = 0.155); however, it was significantly correlated to saliva IgG (n = 12, r = 0.7, P = 0.018) and Brix in serum (n = 12, r = 0.7, P = 0.013). We conclude that IgG was quantifiable in most of the saliva samples. For saliva IgG to be of any value with regards to detecting failure of passive transfer, future studies should investigate methods that can detect IgG <0.1 g/L. The results indicate that saliva IgG can be used to predict serum IgG at levels above 10 g/L, which may warrant further exploration of the use of saliva in the surveillance of failure of passive transfer. The results of the current pilot study did not support the potential usage of a Brix % refractometer to quantify saliva IgG.

Heart rate sensor validation and seasonal and diurnal variation of body temperature and heart rate in domestic sheep.

Advantages of low input livestock production on large pastures, including animal welfare, biodiversity and low production costs are challenged by losses due to undetected disease, accidents and predation. Precision livestock farming (PLF) enables remote monitoring on individual level with potential for predictive warning. Body temperature (Tb) and heart rate (HR) could be used for early detection of diseases, stress or death. We tested physiological sensors in free-grazing Norwegian white sheep in Norway. Forty Tb sensors and thirty HR sensors were surgically implanted in 40 lambs and 10 ewes. Eight (27%) of the HR and eight (20%) of the Tb sensors were lost during the study period. Two Tb sensors migrated from the abdominal cavity in to the digestive system. ECG based validation of the HR sensors revealed a measurement error of 0.2 bpm (SD 5.2 bpm) and correct measurement quality was assigned in 90% of the measurements. Maximum and minimum HR confirmed by ECG was 197 bpm and 68 bpm respectively. Mean passive HR was 90 bpm (SD = 13 bpm) for ewes and 112 bpm (SD = 13 bpm) for lambs. Mean Tb for all animals was 39.6°C (range 36.9 to 41.8°C). Tb displayed 24-hour circadian rhythms during 80.7 % but HR only during 41.0 % of the studied period. We established baseline values and conclude that these sensors deliver good quality. For a wide agricultural use, the sensor implantation method has to be further developed and real-time communication technology added.

Thermography as a Non-Invasive Measure of Stress and Fear of Humans in Sheep.

No data have been published on the use of infrared thermography (IRT) to evaluate sheep emotions. We assessed whether this technique can be used as a non-invasive measure of negative emotions. Two voluntary animal approach (VAA) tests were conducted (and filmed) on five ewes before and after being restrained. The restraining process was performed by a handler for five minutes. IRT was used during restraint and the VAA tests. The lacrimal caruncle temperature was significantly higher during restraint and in the VAA test after the restraint compared with the VAA test before the restraint (Wilcoxon's test; p = 0.04). The latency period until first contact was longer in the second VAA test (132 s) than in the first one (60 s). Our preliminary results suggest that IRT, combined with behavioral data, is a non-invasive technique that can be useful to assess stress and infer about negative emotions in sheep.

Evaluation of Sheep Anticipatory Response to a Food Reward by Means of Functional Near-Infrared Spectroscopy.

Anticipatory behaviour to an oncoming food reward can be triggered via classical conditioning, implies the activation of neural networks, and may serve to study the emotional state of animals. The aim of this study was to investigate how the anticipatory response to a food reward affects the cerebral cortex activity in sheep. Eight ewes from the same flock were trained to associate a neutral auditory stimulus (water bubble) to the presence of a food reward (maize grains). Once conditioned, sheep were trained to wait 15 s behind a gate before accessing a bucket with food (anticipation phase). For 6 days, sheep were submitted to two sessions of six consecutive trials each. Behavioural reaction was filmed and changes in cortical oxy- and deoxy-hemoglobin concentration ([ΔO2Hb] and [ΔHHb] respectively) following neuronal activation were recorded by functional near infrared spectroscopy (fNIRS). Compared to baseline, during the anticipation phase sheep increased their active behaviour, kept the head oriented to the gate (Wilcoxon's signed rank test; p ≤ 0.001), and showed more asymmetric ear posture (Wilcoxon's signed rank test; p ≤ 0.01), most likely reflecting a learnt association and an increased arousal. Results of trial-averaged [ΔO2Hb] and [ΔHHb] within individual sheep showed in almost every sheep a cortical activation during the anticipation phase (Student T-test; p ≤ 0.05). The sheep showed a greater response of the right hemisphere compared to the left hemisphere, possibly indicating a negative affective state, such as frustration. Behavioural and cortical changes observed during anticipation of a food reward reflect a learnt association and an increased arousal, but no clear emotional valence of the sheep subjective experience. Future work should take into consideration possible factors affecting the accurateness of measures, such as probe's location and scalp vascularization.