Correction: Gonçalves Vero et al. Transport of Pigs of Two Market Weights at Two Space Allowances: Effects on Behaviour, Blood Parameters, and Meat Quality under Summer and Winter Conditions. Animals 2023, 13, 2767.

In the original publication [...].

Transport of Pigs of Two Market Weights at Two Space Allowances: Effects on Behaviour, Blood Parameters, and Meat Quality under Summer and Winter Conditions.

This study was carried out through two separate experiments aiming at evaluating the effects of two space allowances (0.54 and 0.62 m2/pig in summer and 0.44 m2/pig and 0.50 m2/pig in winter) on the behavioural and physiological response, and meat quality of pigs of two slaughter weights (120 kg and 140 kg). In summer, higher blood haematocrit levels were found at slaughter in heavier pigs transported at a smaller space allowance (p = 0.04). During lairage, pigs transported at a smaller space allowance started fighting later (p = 0.04). Fighting behaviour was greater in heavier pigs (p ≤ 0.05), whilst their drinking activity was lower (p < 0.05). This resulted in greater exsanguination blood CK levels (p < 0.01) and drier hams (p = 0.05) in heavier pigs. In winter, only lower space allowance influenced some meat quality traits (p < 0.05), but these effects were minor. The effects of space allowance during summer transports on within-truck ambient conditions, post-transport pigs' welfare, and meat quality are similar. Mixing heavier pigs may result in greater aggressiveness and more fatigue-related meat quality variation during summer. Overall, winter transport results may have been biased by the short journey and within-truck load distribution.

Road Transport: A Review of Its Effects on the Welfare of Piglets.

The purpose of this review is to present the best available scientific knowledge on key effects of pre-transport and transport factors influencing the response of piglets to transport stress and post-transport recovery. To date, research on piglet transportation particularly focused on the effects of season (i.e., heat and cold stress), vehicle design features (ventilation type and deck/compartment location), space allowance and transport duration, and piglet genetics. More specifically, in this review the effect of transport duration has been dealt with through its impact on death rate, behaviour and physiological response, and feeling of hunger and thirst. Based on the available literature, clear conclusions can be drawn on the vulnerability of piglets to heat stress during transport. Both short and long transportation have an effect on piglet welfare, with effects being biased by the genetic background, ambient conditions and vehicle design. Further studies investigating the impact of factors such as vehicle design, truck stocking density and environment, piglet genetic background, and weaning age are needed.

Welfare of equidae during transport.

In the framework of its Farm to Fork Strategy, the Commission is undertaking a comprehensive evaluation of animal welfare legislation. This opinion deals with the protection of horses and donkeys during transport. While the opinion focuses primarily on road transport of horses, there are specific sections dealing with the transport of horses on roll-on-roll-off ferries, horses transported by air and the transport of donkeys. In addition, the opinion covers welfare concerns in relation to a specific scenario identified by the European Commission related to the transport of horses on long journeys to slaughterhouses. Current practices related to transport of horses during the different stages (preparation, loading and unloading, transit and the journey breaks) are described. Overall, 13 welfare consequences were identified as being highly relevant for the welfare of horses during transport based on severity, duration and frequency of occurrence: gastro-enteric disorders, handling stress, heat stress, injuries, isolation stress, motion stress, prolonged hunger, prolonged thirst, respiratory disorders, resting problems, restriction of movement, sensory overstimulation and separation stress. These welfare consequences and their animal-based measures are described. A variety of hazards were identified related to factors such as inexperienced/untrained handlers, lack of horse training, structural deficiencies of vehicles/facilities, poor driving skills/conditions, horse separation/regrouping, unfavourable microclimatic and environmental conditions and poor husbandry practices. The opinion contains general and specific conclusions in relation to the different stages of transport. Recommendations to prevent hazards and correct or mitigate welfare consequences have been developed. Recommendations were also developed to define quantitative thresholds for microclimatic conditions within the means of transport and for space allowance. The development of welfare consequences over time was assessed in relation to maximum journey time.

Welfare of pigs during transport.

In the framework of its Farm to Fork Strategy, the Commission is undertaking a comprehensive evaluation of the animal welfare legislation. The present Opinion deals with protection of pigs during transport. The welfare of pigs during transport by road is the main focus, but other means of transport are also covered. Current practices related to transport of pigs during the different stages (preparation, loading/unloading, transit and journey breaks) are described. Overall, 10 welfare consequences were identified as highly relevant for the welfare of pigs during transport based on the severity, duration and frequency of occurrence: group stress, handling stress, heat stress, injuries, motion stress, prolonged hunger, prolonged thirst, restriction of movement, resting problems and sensory overstimulation. These welfare consequences and their animal-based measures are described. A variety of hazards were identified, mainly relating to factors such as mixing of unfamiliar pigs, inappropriate handling methods and devices, the use of pick-up pens, inexperienced/untrained handlers, structural deficiencies of vehicles and facilities, poor driving conditions, unfavourable microclimatic and environmental conditions and poor husbandry practices leading to these welfare consequences. The Opinion contains general and specific conclusions relating to the different stages of transport of pigs. Recommendations to prevent hazards and to correct or mitigate welfare consequences are made. Recommendations were also developed to define quantitative thresholds for microclimatic conditions and minimum space allowance within means of transport. The development of the welfare consequences over time was assessed in relation to maximum journey duration. The Opinion covers specific animal transport scenarios identified by the European Commission relating to transport of cull sows and 'special health status animals', and lists welfare concerns associated with these.

Welfare of small ruminants during transport.

In the framework of its Farm to Fork Strategy, the Commission is undertaking a comprehensive evaluation of animal welfare legislation. The present Opinion deals with the protection of small ruminants (sheep and goats) during transport. The main focus is on welfare of sheep during transport by road but other means of transport and concerns for welfare of goats during transport are also covered. Current practices related to transport of sheep during the different stages (preparation, loading and unloading, transit and journey breaks) are described. Overall, 11 welfare consequences were identified as being highly relevant for the welfare of sheep during transport based on severity, duration and frequency of occurrence: group stress, handling stress, heat stress, injuries, motion stress, predation stress, prolonged hunger, prolonged thirst, restriction of movement, resting problems and sensory overstimulation. These welfare consequences and their animal-based measures are described. A wide variety of hazards, mainly relating to inappropriate or aggressive handling of animals, structural deficiencies of vehicles and facilities, unfavourable microclimatic and environmental conditions and poor husbandry practices, leading to these welfare consequences were identified. The Opinion contains general and specific conclusions in relation to the different stages of transport. Recommendations to prevent hazards and to correct or mitigate welfare consequences have been developed. Recommendations were also developed to define quantitative thresholds for microclimatic conditions within the means of transport and spatial thresholds (minimum space allowance). The development of welfare consequences over time were assessed in relation to maximum journey time. The Opinion covers specific animal transport scenarios identified by the European Commission relating to the export of sheep by livestock vessels, export of sheep by road, roll-on-roll-off vessels and 'special health status animals', and lists welfare concerns associated with these.

Welfare of cattle during transport.

In the framework of its Farm to Fork Strategy, the Commission is undertaking a comprehensive evaluation of the animal welfare legislation. The present Opinion deals with protection of cattle (including calves) during transport. Welfare of cattle during transport by road is the main focus, but other means of transport are also covered. Current practices related to transport of cattle during the different stages (preparation, loading/unloading, transit and journey breaks) are described. Overall, 11 welfare consequences were identified as being highly relevant for the welfare of cattle during transport based on severity, duration and frequency of occurrence: group stress, handling stress, heat stress, injuries, motion stress, prolonged hunger, prolonged thirst, respiratory disorders, restriction of movement, resting problems and sensory overstimulation. These welfare consequences and their animal-based measures are described. A variety of hazards, mainly relating to inexperienced/untrained handlers, inappropriate handling, structural deficiencies of vehicles and facilities, poor driving conditions, unfavourable microclimatic and environmental conditions, and poor husbandry practices leading to these welfare consequences were identified. The Opinion contains general and specific conclusions relating to the different stages of transport for cattle. Recommendations to prevent hazards and to correct or mitigate welfare consequences have been developed. Recommendations were also developed to define quantitative thresholds for microclimatic conditions within the means of transport and spatial thresholds (minimum space allowance). The development of welfare consequences over time was assessed in relation to maximum journey duration. The Opinion covers specific animal transport scenarios identified by the European Commission relating to transport of unweaned calves, cull cows, the export of cattle by livestock vessels, the export of cattle by road, roll-on-roll-off ferries and 'special health status animals', and lists welfare concerns associated with these.

Behavior, blood stress indicators, skin lesions, and meat quality in pigs transported to slaughter at different loading densities.

A total of 1,936 pigs were transported through 8 trips (8.4 ± 0.5 hr) from 2 grow-to-finish farms to a commercial slaughter plant, all located in Southern Brazil. On each trip, a sub-sample of each load (36 barrows/load, weighing 118.9 ± 9.8 kg) was randomly allocated into one of the following loading densities during transport: 200 kg/m2 (D200), 235 kg/m2 (D235), and 270 kg/m2 (D270). Behavioral recordings of postures and activities were made during transportation and lairage using video-cameras. At slaughter, blood samples were collected to assess the concentrations of creatine kinase (CK) and lactate. Carcass weights and skin lesion scores were assessed on-line, and meat quality was evaluated in the longissimus thoracis (LT) muscle by assessing pH, color, and drip loss. During transportation, the proportion of animals lying down was higher (P < 0.05) in D200 and D235 groups compared with D270. The proportion of sitting animals during transportation was higher (P < 0.01) in D270 compared with D200. In lairage, D200 and D235 pigs stood more compared with D270 (P = 0.01), while the proportion of lying pigs was higher (P = 0.02) for D270 pigs compared with D200 and D235. The frequency of drinking bouts in lairage was higher (P < 0.05) for D200 group compared with D235 and D270. The levels of CK were lower (P < 0.05) in D200 pigs transported compared with D270. Lesion scores tended to be higher (P = 0.06) in D270 carcasses compared with D200 and D235. A tendency for lower (P = 0.10) pH1 values in the LT muscle of D270 pigs compared with D200 pigs was also found. Based on the results of this study, the application of lower loading densities (≤235 kg/m2) in the truck allows pigs to have sufficient space to rest, travel more comfortably and arrive less fatigued at the slaughter plant.

Correlation between L-Lactate Concentrations in Beef Cattle, Obtained Using a Hand-Held Lactate Analyzer and a Lactate Assay Colorimetric Kit.

Lactate is a product of anaerobic glycolysis, used in animal research as an indicator of muscle fatigue. Therefore, it has been used as an indicator of cattle response to long distance transportation. The aim of this study was to assess the relationship of L-lactate concentrations measured using a Lactate Scout+ analyzer and a traditional lactate assay colorimetric kit. Blood samples were collected by venipuncture from 96 steers (Black or Red Angus × Hereford/Simmental and Black or Red Angus × Charolais; 247 ± 38.2 kg BW) prior to loading (LO1) and after 36 h of transport, and prior to reloading and after an additional 4 h of road transportation, and on d 1, 2, 3, 5, 14, and 28 after transport. The Lactate Scout+ analyzer strip was dipped in blood at the time of sampling, while blood samples were collected into sodium fluoride tubes for use in the colorimetric analysis. Pearson correlations were calculated to assess the strength of the relationship between the experimental methods for the quantification of L-lactate concentrations. The magnitude and direction of the correlation, and the level of statistical significance varied over the observed time points, ranging from r = -0.03 (p = 0.75; LO1) to r = 0.75 (p < 0.0001; d 3). The correlation for the pooled data was weak but statistically significant (r = 0.33, p < 0.0001). Based on the low magnitude of the correlation due to variability across sampling time points in this study, the Lactate Scout+ analyzer is not a suitable alternative to a lab-based assay (considered the gold standard) for measuring L-lactate in transported cattle.

The effects of feeding finishing pigs of two genders with a high fiber and high fat diet on muscle glycolytic potential at slaughter and meat quality.

A total of 160 pigs, in groups of 8 pigs of mixed genders, were fed four finishing feeding strategies with the aim to reduce muscle glycolytic potential and improve meat quality. Pigs were fed a control diet (C; fat = 5.0%, ADF = 3.0%, NDF = 8.8%), a high-fat and high-fiber diet (HFF; fat = 11.2%, ADF = 9.1%, NDF = 19.5%), a blend of 50-50% C and HFF diets (fat = 8.2%, ADF = 6.7%, NDF = 14.2%) or the C diet and transferred to the HFF diet after a diet transition. Dietary treatments alone or in interaction with gender had no effect on pig growth performance, carcass quality traits, Longissimus and Semimembranosus muscle glycolytic potential and meat quality (P > 0.10). The inefficiency of the dietary treatments applied in this study may be due to the low ratio between fat and digestible carbohydrate in the diets combined with the mild pre-slaughter stress conditions pigs were exposed to.

Effects of energy supplements on the differentiation of skeletal muscle satellite cells.

To investigate the effects of the activator of AMPK and high glucose on the differentiation of mouse SMSCs, primary SMSCs were isolated from mouse extensor digitorum longus muscle and grown to near confluence (80%). Postconfluent cells were cultured in a growth medium with different inductors: AICAR, glucose, and AICAR mixed with glucose. The specific protein expressions of SMSCs, myoblasts, adipocytes, and brown adipocytes were analyzed on days 0, 3, 5, 7, and 10. The results showed treatment with AICAR in SMSCs markedly activated AMPK phosphorylation (p < .05) and increased protein expression of Pax7 and MyoD (p < .05), high concentrations of intracellular glucose upregulated UCP-1 protein expression and enhanced lipid accumulation (p < .05), the cowork of AICAR and glucose affected a decrease on MyoD, PPARg, and UCP-1 expression (p < .05) and an increase on Pax7. The present study indicated that the certain energy supplements influence the direction of SMSC differentiation which may contribution on the structure of muscle and meat quality, sequentially.

Association Between Tail-Biting and Intestinal Microbiota Composition in Pigs.

Tail-biting (TB) in pigs is a serious behavioral disorder. It is an important challenge in swine production as it impacts animal welfare and health and the economics and safety of the pork meat supply chain. To prevent TB, approaches including enrichment material and tail docking are proposed but none are optimal. Nutrition appears to be an important factor in TB behavior, perhaps by modulating the intestinal microbiota (IM). Our aim was to assess the association between TB behavior and IM in pigs through comparisons of IM in groups of biter, bitten and non-biter/non-bitten pigs. Each group composed of 12 pigs was formed at the beginning of the growing/finishing phase based on a target behavior analysis centered on TB behavior for the biter group and a score of damages caused to the tail for the bitten group. Blood and fecal samples were collected from each pig during a TB episode, at time 0, t0, and when the TB episode was considered finished, 4 weeks later, at time 1, t1. Serum cortisol level was determined by ELISA and used as an indicator of stress. The pig's fecal microbiota was analyzed from DNA extracted from freshly collected fecal matter using amplicon sequencing of the V4 hypervariable region of the 16S rRNA gene. Serum cortisol levels were significantly higher in either the biter or bitten pig groups compared to the negative control group (p = 0.02 and p = 0.01, respectively). The microbiota alpha-diversity was not significantly different between all groups, biter, bitten and negative control. Analyses of beta-diversity, however, revealed a significant difference between either the biter or the bitten group in comparison to the non-biter/non-bitten negative control group in terms of structure and composition of the microbiota. Lactobacillus were significantly more abundant in the negative control group compared to the two other groups (p = 0.001). No significant difference was revealed between the biter and bitten groups. Quantitative real-time PCR (qPCR) confirmed that lactobacilli were more abundant in the negative control group. Our study indicates that TB behavior is associated with the IM composition in pigs.

Evaluation of physical euthanasia for neonatal piglets on-farm.

Twenty-seven neonate piglets (range from 0.35 to 1.17 kg) were evaluated for the effectiveness of blunt force trauma as a method of on-farm cull. Brainstem function, brain injury, and hemorrhage scores (increasing from 0 to 3) were assessed after striking the head against a concrete floor. Electroencephalograms (EEG) from a subset of 15 piglets were recorded before and after blunt force trauma for electrophysiological assessments. Blunt force trauma was performed by a single experienced farmer in a commercial farm by holding the piglet by its both hind legs and striking the head against the concrete floor. All piglets remained recumbent and did not show brainstem reflexes. Only one piglet did not presented tonic/clonic physical activity. The mean time to the onset of persistent isoelectric EEG was 64.3 ± 7.3 s (range 18 to 115). Total power, theta, alpha, and beta power decreased to approximately 45%, 30%, 20%, and 15% from pretreatment power, respectively, by 15-s post-impact. There were no periods of normal-like EEG after the culling. Bruises in the neck and shoulder were found in 67% and 70% of piglets, respectively. All piglets presented skull fractures with 20% having the nasal bone(s) fractured. Brain damage was found in all piglets, mainly in the frontal lobe(s). The occipital lobe(s) presented the greatest frequency of severe damage. The analysis of the radiographs also found a high frequency of fractures in this region. Hemorrhage was most frequent in the frontal, parietal, occipital lobes, and midbrain. When performed correctly with the appropriate weight class, blunt force trauma can be used as an effective method for the on-farm killing of nursing piglets resulting in death. However, this method should not be promoted over more reliable and repeatable cull methods such as captive bolt gun. As with blunt force trauma, there is a significant potential for animal welfare harm associated with inappropriate practice, lack of accuracy, issues with repeatability, and operator fatigue.

Application of extended feed withdrawal time preslaughter and its effects on animal welfare and carcass and meat quality of enriched-housed pigs.

A total of 144 barrows were distributed into four treatment groups. One group was housed conventionally and fasted for 16 h preslaughter, while the other three groups were housed in enriched conditions (straw-bedding and low density) and fasted for 16, 24 and 32 h preslaughter. When compared with conventionally-housed pigs, enriched-housed pigs were heavier (P < .05) and fatter (P < .01), but easier to handle at loading (P < .05) and produced less exudative pork (P < .05) possibly resulting from a lower post-mortem muscle glycolytic potential (P = .10). Extending fasting time up to 32 h reduced dressing yield (P < .001) and increased skin lesion scores (P < .01), but did not result in any clear or major effect on muscle metabolism and meat quality. In conclusion, given the beneficial effects of enriched housing conditions on stress response and pork quality, the application of an extended fasting time (32h) preslaughter is not required for this production.

Validation of Anatomical Sites for the Measurement of Infrared Body Surface Temperature Variation in Response to Handling and Transport.

This study aimed at validating the anatomical sites for the measurement of infrared (IR) body surface temperature as a tool to monitor the pigs' response to handling and transport stress. The selected anatomical sites were the neck (infrared neck temperature-IRNT), rump (infrared rump temperature-IRRT), orbital (infrared orbital temperature-IROT) and behind ears (infrared behind ears temperature-IRBET) regions. A total of 120 pigs were handled from the finishing pen to the loading dock through a handling test course. Two handling types (gentle vs. rough) and number of laps (1 vs. 3) were applied according to a 2 × 2 factorial design. After loading, pigs were transported for 40 min and returned to their home pens. Animal behavior, heart rate, rectal temperature and salivary cortisol concentration were measured for validation. Increased IR body temperature, heart rate and salivary cortisol levels were observed in response to rough handling and longer distance walk (P < 0.05 for all). The greatest correlations were found between IROT and IRBET temperatures and salivary cortisol concentration at the end of the handling test (r = 0.49 and r = 0.50, respectively; P < 0.001 for both). Therefore, IR pig's head surface temperature may be useful for a comprehensive assessment of the physiological response to handling and transport stress.

A Review of Swine Transportation Research on Priority Welfare Issues: A Canadian Perspective.

The purpose of this review is to present the best available scientific knowledge on key animal welfare issues during swine transport, such as transport duration and distance, time off feed and water, rest intervals, environmental conditions, loading density, and transport of young animals, based on their impact on stress, injury, fatigue, dehydration, body temperature, mortality, and carcass and meat quality. The review was limited to this set of priority welfare issues which were identified by the National Farm Animal Care Council (NFACC) Scientific Committee to help with the development of the livestock transportation Codes of Practice. This review focuses primarily on research related to the transport of market pigs (100-135 kg) which is a reflection of the current literature available on pig transportation. This information presented here can be used to support other animal welfare codes, guidelines, standards or legislations regulating the welfare of pigs during transport. Based on the available literature, clear conclusions can be drawn on the impact of vehicle design, pre-transport fasting, control of environmental conditions and loading density on the welfare of pigs during transport and on pork quality. However, the effects of journey duration are still unclear and a recommendation on the maximum transport time cannot be provided. Further studies investigating the impact of factors, such as ambient conditions within the transport vehicle, loading density at extreme ambient conditions, travel distances, maximum travel duration, rest/stop duration, and management of pigs during rest stops are required. More specifically, further research in relation to the welfare of market weight, newly weaned and breeding pigs, and cull sows and boars during transport is needed.

Effects of Transport Duration and Environmental Conditions in Winter or Summer on the Concentrations of Insulin-Like Growth Factors and Insulin-Like Growth Factor-Binding Proteins in the Plasma of Market-Weight Pigs.

In previous work using market-weight pigs, we had demonstrated that insulin-like growth factors (IGFs) and insulin-like growth factor binding proteins (IGFBPs) are regulated during shipment characterized by changing conditions of stress due to loading or unloading, transportation, lairage, and slaughter. In addition, we found in a previous study that IGFBP-2 concentrations were lower in pigs transported for longer periods of time. Therefore, we performed a more detailed study on the effects of transport duration and season on the plasma concentrations of IGFs and IGFBPs in adult pigs. For the study, exsanguination blood was collected from 240 market-weight barrows that were transported for 6, 12, or 18 h in January or July. IGF-I and -II were detected using commercial ELISAs whereas IGFBPs were quantified by quantitative Western ligand blotting. In addition, established markers of stress and metabolism were studied in the animals. The results show that plasma concentrations of IGFBP-3 were significantly reduced after 18 h of transport compared to shorter transport durations (6 and 12 h; p < 0.05). The concentrations of IGF-I in plasma were higher (p < 0.001) in pigs transported 12 h compared to shorter or longer durations. Season influenced plasma concentrations of IGFBP-3 and IGF-II (p < 0.05 and p < 0.01, respectively). Neither transport duration nor differential environmental conditions of winter or summer had an effect on glucocorticoids, albumin, triglycerides, or glucose concentrations (p > 0.05). However, low-density lipoprotein concentrations decreased after 18 h compared to 6 h of transport (p < 0.05), whereas high-density lipoprotein concentrations were higher (p < 0.05) in pigs transported for 12 or 18 h compared to those transported for only 6 h. Our findings indicate differential regulation of IGF-compounds in response to longer transport duration or seasonal changes and support current evidence of IGFs and IGFBPs as innovative animal-based indicators of psycho-social or metabolic stress in pigs.

Preslaughter handling practices and their effects on animal welfare and pork quality.

At all times, prior to slaughter, pigs may experience stress from a range of handling practices, such as fasting, loading and transport, mixing, and interaction with humans. These factors can affect the welfare of pigs and carcass and meat quality, both individually and collectively. Preslaughter stress is both an animal welfare and a meat quality issue. Behavioral and physiological studies have revealed that poor handling practices at the farm, during transport and at the slaughter plant, have an adverse effect on pigs and may result in the loss of profits due to animal losses during transport and in lairage. Also, poor preslaughter handling can also lead to losses in carcass value as a result of reduced yield, the presence of lesions and bacterial contamination, and meat quality defects (e.g., pale, soft, exudative and dark, firm, dry pork). These economic losses can be limited by improving the design of facilities, controlling the environmental conditions, and implementing training programs for the correct animal handling at any stage preslaughter. The objective of this review was to review research findings on the effects of preslaughter practices on ante-mortem behavioral and physiological response in pigs, including muscle metabolism, and to provide recommendations aimed at limiting the impact of preslaughter handling on animal losses and pork quality.

Interference of stress with the somatotropic axis in pigs - lights on new biomarkers.

The acceptance of animal products is increasingly associated with standardized animal welfare, which relates to appropriate animal husbandry from birth to slaughter. In particular, shipment to the slaughterhouse is considered as a critical process exposing the animals to a number of, in part severe, stressors. New biomarkers may be useful for the assessment of animal welfare. The IGF-system has been assessed in a commercial pig transport in conjunction with established markers of stress response. Furthermore, the effect of repeated restraint as an experimental model for repeated acute stress was investigated. During shipment from farm to slaughterhouse, plasma concentrations of IGFBP-3 and IGFBP-2 were significantly reduced (p < 0.01). After shipment, the plasma concentrations of IGFBP-5, glucocorticoids and IL-2 increased but decreased after lairage (p < 0.05) whereas IGF-1 decreased after shipment (p < 0.01). Repeated acute stress increased concentrations of IGFBP-3 and IGF-1 in exsanguination blood (p < 0.05). Differential IGF- signatures can indicate altered endocrine or metabolic control and thus contain complex animal-related information. The somatotropic axis may be of particular interest when established biomarkers such as cortisol, glucose, or lactate cannot be used for the assessment of animal stress or welfare.

Can the way pigs are handled alter behavioural and physiological measures of affective state?

Research on human-animal relationship in animal production has been mainly focused on its effect on stress, productivity and meat quality. Only few studies have assessed its effects on the animals' affective state. In the present study, the influence of positive and negative handling (pH and NH, respectively) on affective state and fear as assessed by the cognitive bias test, the novel object test and the defence cascade test was studied in 56 pigs. Serum, saliva and hair were sampled during the study for the analysis of cortisol concentration. Results showed no differences between pH and NH pigs in the behavioural tests, which may be either due to the lack of previous handling effect on the test results, the lack of validity or the low sensitivity of these tests or a combination of them. Moreover, no differences were found in cortisol concentrations between handling groups. However, correlations between tests were found (p<0.05) suggesting that there are individual factors such as the fear level, the motivation or the coping style, that have a similar effect on the response to these tests. Moreover, pigs who were more fearful had higher (r=0.37; p=0.014) levels of serum cortisol at slaughter.