Neurofilament light chain levels in pregnant multiple sclerosis patients: a prospective cohort study.

BACKGROUND AND PURPOSE: Neurofilament light chain is a cytoskeletal protein of neurons. Its levels are increasingly recognized as measures of neuroaxonal damage. The aim of this study was to explore serum neurofilament light chain (sNfL) levels in multiple sclerosis (MS) patients and healthy controls during pregnancy and puerperium. METHODS: This was a prospective, longitudinal, single-center study. sNfL concentration was assessed using a highly sensitive single-molecule array during pregnancy and in puerperium, in a cohort of 39 pregnant patients with relapsing multiple sclerosis (P-MS). Twenty-one healthy pregnant women (HPW) served as a control group. Eight P-MS suffered relapses during pregnancy (P-MS-R) in the first or second trimesters. RESULTS: No differences in pregnancy and delivery data were observed between P-MS and HPW. P-MS showed higher sNfL values than HPW in the first trimester, independently of the presence (P = 0.002) or not (P = 0.02) of relapses during pregnancy. However, in the third trimester, only P-MS-R showed higher sNfL values than HPW (P = 0.001). These differences extended to the puerperium, where P-MS-R showed higher sNfL values than those with no relapses during gestation (P = 0.02). CONCLUSION: These data strongly suggest that sNfL levels reflect MS activity during pregnancy. Additionally, the absence of relapses during pregnancy may have a beneficial effect on neurodegeneration during puerperium.

Clinical usefulness of prognostic biomarkers in optic neuritis.

BACKGROUND AND PURPOSE: Different biological and radiological biomarkers predict clinical conversion to multiple sclerosis (MS) after a clinically isolated syndrome (CIS). The aim was to explore their role in predicting the outcome of patients with optic neuritis (ON), a CIS considered to have a benign prognosis. METHODS: Sixty-eight consecutive ON patients were followed prospectively. Magnetic resonance imaging (MRI) and cerebrospinal fluid studies including oligoclonal immunoglobulin G (IgG) bands (OCGBs), lipid-specific oligoclonal IgM bands (LS-OCMBs) and neurofilament light chain quantification were performed at disease onset. Conversion to clinically definite MS (CDMS) was monitored. RESULTS: The mean time of follow-up of our series was 46.4 months. Twenty-five patients (36.7%) developed CDMS during follow-up. Neurofilament light chain levels did not predict clinical conversion. By contrast, an abnormal MRI increased the risk of CDMS [hazard ratio (HR) 12.5, P = 0.013]. The clearest association was found in patients with more than three T2 lesions. OCGBs also predicted the onset of CDMS (HR 21.3, P = 0.003) and LS-OCMBs were associated with a shorter time to CDMS (HR = 116.6, P < 0.001). CONCLUSIONS: Magnetic resonance imaging and OCGBs predicted conversion to CDMS after an ON episode. In addition, LS-OCMBs identified the ON patients more likely to develop MS early. These results, applicable to the everyday clinical setting, may be of interest for therapeutic decisions.

Pig cognitive bias affects the conversion of muscle into meat by antioxidant and autophagy mechanisms.

Slaughter is a crucial step in the meat production chain that could induce psychological stress on each animal, resulting in a physiological response that can differ among individuals. The aim of this study was to investigate the relationship between an animal's emotional state, the subsequent psychological stress at slaughter and the cellular damage as an effect. In all, 36 entire male pigs were reared at an experimental farm and a cognitive bias test was used to classify them into positive bias (PB) or negative bias (NB) groups depending on their decision-making capabilities. Half of the animals, slaughtered in the same batch, were used for a complete study of biomarkers of stress, including brain neurotransmitters and some muscle biomarkers of oxidative stress. After slaughter, specific brain areas were excised and the levels of catecholamines (noradrenaline (NA) and dopamine (DA)) and indoleamines (5-hydroxyindoleacetic acid and serotonin (5HT)) were analyzed. In addition, muscle proteasome activity (20S), antioxidant defence (total antioxidant activity (TAA)), oxidative damage (lipid peroxidation (LPO)) and autophagy biomarkers (Beclin-1, microtubule-associated protein I light chain 3 (LC3-I) and LC3-II) were monitored during early postmortem maturation (0 to 24 h). Compared with PB animals, NB pigs were more susceptible to stress, showing higher 5HT levels (P<0.01) in the hippocampus and lower DA (P<0.001) in the pre-frontal cortex. Furthermore, NB pigs had more intense proteolytic processes and triggered primary muscle cell survival mechanisms immediately after slaughter (0 h postmortem), thus showing higher TAA (P<0.001) and earlier proteasome activity (P<0.001) and autophagy (Beclin-1, P<0.05; LC3-II/LC3-I, P<0.001) than PB pigs, in order to counteract the induced increase in oxidative stress, that was significantly higher in the muscle of NB pigs at 0 h postmortem (LPO, P<0.001). Our study is the first to demonstrate that pig's cognitive bias influences the animal's susceptibility to stress and has important effects on the postmortem muscle metabolism, particularly on the cell antioxidant defences and the autophagy onset. These results expand the current knowledge regarding biomarkers of animal welfare and highlight the potential use of biomarkers of the proteasome, the autophagy (Beclin-1, LC3-II/LC3-I ratio) and the muscle antioxidant defence (TAA, LPO) for detection of peri-slaughter stress.

Urgent request on avian influenza.

Highly pathogenic avian influenza (HPAI) H5N8 is currently causing an epizootic in Europe, infecting many poultry holdings as well as captive and wild bird species in more than 10 countries. Given the clear clinical manifestation, passive surveillance is considered the most effective means of detecting infected wild and domestic birds. Testing samples from new species and non-previously reported areas is key to determine the geographic spread of HPAIV H5N8 2016 in wild birds. Testing limited numbers of dead wild birds in previously reported areas is useful when it is relevant to know whether the virus is still present in the area or not, e.g. before restrictive measures in poultry are to be lifted. To prevent introduction of HPAIV from wild birds into poultry, strict biosecurity implemented and maintained by the poultry farmers is the most important measure. Providing holding-specific biosecurity guidance is strongly recommended as it is expected to have a high impact on the achieved biosecurity level of the holding. This is preferably done during peace time to increase preparedness for future outbreaks. The location and size of control and in particular monitoring areas for poultry associated with positive wild bird findings are best based on knowledge of the wider habitat and flight distance of the affected wild bird species. It is recommended to increase awareness among poultry farmers in these established areas in order to enhance passive surveillance and to implement enhanced biosecurity measures including poultry confinement. There is no scientific evidence suggesting a different effectiveness of the protection measures on the introduction into poultry holdings and subsequent spread of HPAIV when applied to H5N8, H5N1 or other notifiable HPAI viruses.

One Welfare - a platform for improving human and animal welfare.

Adopting the concept of One Welfare could help to improve animal welfare and human wellbeing worldwide, argue Rebeca García Pinillos, Michael Appleby, Xavier Manteca, Freda Scott-Park, Charles Smith and Antonio Velarde.

Can the monitoring of animal welfare parameters predict pork meat quality variation through the supply chain (from farm to slaughter)?

The objective of this study was to assess the relationship between the animal welfare conditions evaluated through the supply chain and pork quality variation. A total of 4,680 pigs from 12 farms-5 animal welfare improved raising system (AWIRS) and 7 conventional raising system (CON) farms-were assessed from farm to slaughter through a comprehensive audit protocol merging the European Welfare Quality, the Canadian Animal Care Assessment, and American Meat Institute audit guide criteria. At the abattoir, a subsample of 1,440 pigs (120 pigs/farm) was randomly chosen out of 24 loads (2 farms per wk) transported by 2 drivers (driver A and driver B) for the assessment of stunning effectiveness, carcass bruises, blood lactate levels, and meat quality traits. Meat quality was assessed in the longissimus lumborum (LL) muscle 24 h postmortem by measuring ultimate pH (pHu), color (L*, a*, and b*), and drip loss. Data were analyzed by the MIXED, GLIMMIX, and NAPAR1WAY procedures of SAS. Spearman correlations were calculated to determine the relationship between audit scores and meat quality traits. Better animal welfare conditions, as showed by greater final scores for good housing (GHo; = 0.001) and good health ( = 0.006) principles, were recorded at AWIRS farms. Pigs from AWIRS farms handled by driver B displayed a greater percentage of turning back ( = 0.01) and slips ( < 0.001) during unloading and a greater ( = 0.02) frequency of falls in the stunning chute. A greater ( = 0.02) reluctance to move at loading was found in CON pigs loaded by driver A compared with driver B, whereas a greater ( < 0.001) reluctance to move was found in these pigs at unloading when they were unloaded by driver B. Drip loss was higher ( = 0.003) and pale, soft, and exudative pork percentage was greater ( < 0.001) in the LL muscle of the heavier AWIRS pigs. The GHO principle was best correlated with pHu ( = -0.75, = 0.01) and Minolta L* value ( = 0.87, < 0.001) of the LL muscle. Overall, drip loss variation in the LL muscle was correlated with the frequency of slips at unloading ( = 0.63, = 0.001) and in the restrainer area ( = 0.74, < 0.001). The results of this study showed that the quality of the raising system and truck driver skills as assessed by animal welfare audit protocols are important sources of variation in the behavioral response of pigs to preslaughter handling and may affect pork quality variation. However, the different live weight between CON and AWIRS pigs may have biased the meat quality results in this study.

Development of a new welfare assessment protocol for practical application in long-term dog shelters.

In many European shelters, dogs may spend many years confined. A poor environment and inappropriate management may lead to a low quality of life. The absence of harmonised European regulatory frameworks defining the minimum requirements for shelter facilities makes the definition of welfare standards for kennelled dogs challenging. Here, a new protocol was developed and tested to help identify the main welfare issues for shelter dogs. Twenty-six indicators were identified including management, resource and animal based measures. Accuracy and interobserver reliability were checked between four assessors. The protocol was applied in 29 shelters (n=1308 dogs) in six European countries. Overall prevalence of poor health conditions was below 10%. Test-retest reliability and validity of the protocol were investigated with encouraging results. A logistic regression was carried out to assess the potential of the protocol as a tool to identify welfare hazards in shelter environments. Inappropriate space allowance, for example, was found to be a risk factor potentially affecting the animal's cleanliness, skin condition and body condition. The protocol was designed to be concise and easy to implement. Systematic data collection could help identify welfare problems that are likely to arise in certain shelter designs and thus determine improvement in animal care standards.

Comparison of behaviour, performance and mortality in restricted and ad libitum-fed growing rabbits.

The objective of this study was to determine whether rabbits fed in a restricted regimen (75%) showed increased competition for feeding, drinking and use of specific areas of the cages as compared with those provided feed ad libitum. This evaluation was carried out by measuring their space utilisation in the cage, the incidence of agonistic behaviour and rates of mortality. In total, 504 rabbits between 31 and 66 days of age were used in this study. A total of 200 heavy-weight rabbits and 56 light-weight rabbits were randomly housed in 32 cages, each cage containing eight rabbits: 25 cages housing heavy rabbits and seven cages housing the light-weight ones. They were all fed ad libitum (AD). In addition, a total of 208 heavy-weight rabbits and 40 light-weight rabbits were randomly housed in 31 cages, each of them containing eight rabbits: 26 cages housing heavy weight rabbits and five cages housing light-weight ones. They were all fed a restricted diet (R) regimen. The restriction was calculated to be 75% of the feed consumed by the AD group. The total space available in the cage was 3252 cm(2), with a stocking density of 24.6 animals/m(2). Animals between 32 and 60 days of age from 20 different cages were observed nine times per week (morning or afternoon) by means of scan and focal sampling by one observer. During each period, cages were assessed for 5 min, registering every minute the position of all the animals in relation to Area A (feeder), Area B (central part) or Area C (back and drinker area). The incidence of agonistic behaviour such as displacement, biting and jumping on each other was also assessed. Performance variables such as daily gain and feed conversion ratio, in addition to general health status and mortality rates, were recorded for all rabbits. When the rabbits were under restricted feeding, the competition for feed and drink increased with clear signs of agonistic behaviour such as biting, displacement and animals jumping on top of each other. Although this competition was maintained during the entire growing period, the BW homogeneity between animals in the same cage was similar in both cases, suggesting that all animals could consume similar quantities of feed. The possible advantages of a restricted diet, such as better feed conversion ratio, were observed in this study only in the last few weeks of the growing period.

Use of an anti-GnRF vaccine to suppress estrus in crossbred Iberian female pigs.

Antibodies against GnRF elicited by vaccination with Improvac are expected to have the same effects on the gonads of female pigs as those observed in males and thus suppress estrus. The objective of this study was to evaluate whether a 3- or a 4-dose regimen of Improvac was effective in inducing and maintaining suppression of estrus in intact Iberian female pigs through 14 months of age. Sixty 18-week-old gilts were randomly assigned to one of the following three treatment groups: control (PBS injected), V3 (3-dose Improvac regimen), and V4 (4-dose Improvac regimen). Animals were tested for the presence of standing estrus in the presence of a boar three times per week until study completion at 60 weeks of age. Blood samples were regularly collected to determine the serum levels of anti-GnRF antibodies and progesterone. The weight and size of the uterus and the weight and activity of the ovaries were assessed at the time of euthanasia. For both treatment groups V3 and V4, a large reduction in the incidence of standing estrus was observed when compared to the control group (P < 0.0001). No significant differences between the V3 and V4 groups were observed. Both V3 and V4 groups had higher serum anti-GnRF antibody levels than the control group from 28 days after the first vaccination until the end of the study. In addition, both V3 and V4 groups had lower levels of progesterone than the control group from Day 112 until the end of the study (P < 0.0001 in all cases). Mean ovary weights at study end in treatment groups V3 and V4 were significantly lower (P < 0.0001) than those for the control group and not significantly different between the V3 and V4 groups. An onset of immunity of 2 weeks after the second vaccination and a long-lasting immunity of at least 20 weeks after the third vaccination were observed. In summary, both the 3-and 4-dose Improvac regimens were efficacious in reducing the incidence of standing estrus, serum progesterone levels, and the development of the uterus and ovaries.

Effect of animal mixing as a stressor on biomarkers of autophagy and oxidative stress during pig muscle maturation.

The objective of this work was to study the postmortem evolution of potential biomarkers of autophagy (Beclin 1, LC3-II/LC3-I ratio) and oxidative stress (total antioxidant activity, TAA; superoxide dismutase activity, SOD and catalase activity, CAT) in the Longissimus dorsi muscle of entire male ((Large White×Landrace)×Duroc) pigs subjected to different management treatments that may promote stress, such as mixing unfamiliar animals at the farm and/or during transport and lairage before slaughter. During the rearing period at the farm, five animals were never mixed after the initial formation of the experimental groups (unmixed group at the farm, UF), whereas 10 animals were subjected to a common routine of being mixed with unfamiliar animals (mixed group at the farm, MF). Furthermore, two different treatments were used during the transport and lairage before slaughter: 10 pigs were not mixed (unmixed group during transport and lairage, UTL), whereas five pigs were mixed with unfamiliar animals on the lorry and during lairage (mixed group during transport and lairage, MTL). These mixing treatments were then combined into three pre-slaughter treatments - namely, UF-UTL, MF-UTL and MF-MTL. The results show that MF-UTL and MF-MTL increased significantly the muscle antioxidant defense (TAA, SOD and CAT) at short postmortem times (4 and 8 h; P<0.001), followed by an earlier depletion of the antioxidant activity at 24 h postmortem (P<0.05). We also found that mixing unfamiliar animals, both at the farm and during transport and lairage, triggers postmortem muscle autophagy, which showed an earlier activation (higher expression of Beclin 1 and LC3-II/LC3-I ratio at 4 h postmortem followed by a decreasing pattern of this ratio along first 24 h postmortem) in the muscle tissues of animals from the MF-UTL and MF-MTL groups, as an adaptive strategy of the muscle cells for counteracting induced stress. From these results, we propose that monitoring the evolution of the main biomarkers of autophagy (Beclin 1, LC3-II/LC3-I ratio) and muscle antioxidant defense (TAA, SOD, CAT) in the muscle tissue within the first 24 h postmortem may help the detection of animal stress and its potential effect on the postmortem muscle metabolism.

Electrical stunning effectiveness with current levels lower than 1 A in lambs and kid goats.

An experiment with 360 lambs grouped into three Spanish commercial categories, (Pascual, 13-16 kg; Recental, 9-13 kg and Lechal <7 kg carcass weight) and kid goats (7 kg) was performed to assess stunning effectiveness after head-only (HO) and head-to-body (HB) electrical stunning with intensity currents of 0.3, 0.5 and 0.7 Amperes (A) compared to 1.0 A. After stunning, all animals showed tonic-clonic muscular activity and epileptiform EEG, absence of rhythmic breathing, corneal reflex, spontaneous blinking and pain sensibility. The quiescent EEG occurred earlier (P < 0.05) in HB compared to HO in all categories. More animals recovered corneal reflex and rhythmic breathing before onset of the quiescent activity after HO (from 15 to 50%) compared to HB (from 0 to 15%) (P < 0.05). Concluding, HO and HB electrical stunning with 0.3, 0.5 and 0.7 A induce effective stunning similar to 1.0 A in lambs and kid goats. After stunning and sticking, brain failure occurs earlier in HB than HO system.

The effects of lairage time and handling procedure prior to slaughter on stress and meat quality parameters in pigs.

Lairage time (short - 8min to 2.7h, n=28 vs. long - 14 to 21.5h, n=72) and pig handling (gentle - no use of stick or electric prod, pig not slipping, falling, nor emitting high-pitched vocalizations vs. rough - where any of these occurred) effects on pig stress and meat quality were measured. Blood lactate and cortisol, plus post-mortem pH (pH60min; pH24h), temperature (T60min), drip loss, sensory and instrumental color and meat quality for the longissimus dorsi, pars lumbalis derived meat were determined. Carcass rigor mortis and skin damages were measured. Lairage time significantly affected blood lactate, carcass rigor mortis, skin damages, drip loss, color and meat quality. Handling procedure influenced blood lactate, pH60min and T60min. Long lairage was more stressful, and was detrimental to carcass quality, but caused better meat quality compared to short lairage. Rough handling was related to higher lactate and lower meat quality.

Religious slaughter: evaluation of current practices in selected countries.

As part of the project "Religious slaughter (DIALREL): improving knowledge and expertise through dialogue and debate on issues of welfare, legislation and socio-economic aspects", this paper discusses an evaluation of current practices during Halal and Shechita slaughter in cattle, sheep, goats and poultry. During religious slaughter, animals are killed with and without stunning by a transverse incision across the neck that is cutting the skin, muscles (brachiocephalic, sternocephalic, sternohyoid, and sternothyroid), trachea, esophagus, carotid arteries, jugular veins and the major, superficial and deep nerves of the cervical plexus. In this report, the restraint methods, stunning, neck cutting, exsanguination, slaughter techniques and postcut handling in the abattoir were assessed for religious slaughter. Information about the procedures used during religious slaughter in Belgium, Germany, Italy, the Netherlands, Spain, the UK, Turkey and Australia was collected by means of spot visits to abattoirs. To standardize the information gathered during the spot visits three guidelines were designed, one for each species, and translated into the national languages of the countries involved. The document included questions on the handling and restraint methods (stunning, neck cutting/exsanguination/slaughter techniques and postcut handling performed under religious practices) and for pain and distress of the animal during the restraint, neck cutting and induction to death in each abattoir. Results showed differences in the time from restraining to stun and to cut in the neck cutting procedures and in the time from cut to death.

Use of infrared thermography to assess the influence of high environmental temperature on rabbits.

The aim of this work was to ascertain if infrared thermography (IRT) can be used on rabbits to assess differences in surface body temperature when they are subjected to two different environmental temperatures outside the comfort zone. Rabbits housed in room A were maintained at a temperature of below 30°C and rabbits in room B at a temperature of above 32°C for a year. Faeces were collected six times during the year to assess stress by means of faecal cortisol metabolites (FCM). The assessment of IRT was carried out to assess maximum and minimum temperatures on the eyes, nose and ears. FCM concentration was higher in room B than A, to confirm that stress conditions were higher in room B. Significant differences in IRT were found between the animals housed in both rooms. It was observed that it was more difficult for animals from room B to maintain a regular heat loss. Although all the body zones used to assess temperature with IRT gave statistical differences, the correlations found between the eyes, nose and ears were moderate, suggesting that they were giving different information. In addition, differences up to 3.36°C were found in the eye temperature of rabbits housed in the same room, with a clear effect of their position in relation to extractors and heating equipments. Therefore, IRT could be a good tool to assess heat stress in animals housed on typical rabbit farm buildings, giving a measure of how the animal is perceiving a combination of humidity, temperature and ventilation. Some face areas were better for analysing images. Minimum temperature on eyes and temperatures on nose are suggested to assess heat losses and critical areas of the farm for heat stress in rabbits.

Influence of social dominance on production, welfare and the quality of meat from beef bulls.

The aim of this study was to assess the effect of social dominance on some indicators of welfare, production and meat quality of young bulls. A total of 60 bulls of the Gasconne breed, 9 months old, housed indoors were used. Indices of success order were calculated to reflect social dominance of each bull into three ranking categories (low, middle and high). Blood samples were taken to measure cortisol, lactate, glucose, creatine kinase, non-esterified fatty acid and neutrophil/lymphocyte ratio (N/L). M. longissimus samples were analysed in terms of pH, water holding capacity (WHC), texture, colour and sensorial attributes. Social rank influenced cattle stress response, which had an effect on productive performance but not on meat quality traits, with the exception of the sensory traits. These results emphasize the importance of implementing best management practices during pre-harvest handling of cattle in order to modulate any possible risk factor for social stress.

Assessment of unconsciousness in pigs during exposure to nitrogen and carbon dioxide mixtures.

The aim of this study was to assess unconsciousness in pigs during and after the exposure to gas mixtures of 70% nitrogen (N(2)) and 30% carbon dioxide (CO(2)) (70N30C), 80% N(2) and 20% CO(2) (80N20C) and 85% N(2) and 15% CO(2) (85N15C) compared with 90% CO(2) in air (90C) by means of the Index of Consciousness(®)(IoC), their behaviour and the absence of brain stem reflexes. The experiment included three trials of 24 pigs divided into four groups according to the number of treatments. Half of the group was exposed for a short time and the other half for a long time (3 and 5 min for the N(2)/CO(2) mixtures exposure and 2 and 3 min in 90C exposure, respectively). During exposure, the IoC and the electroencephalography suppression rate (ESR) were assessed, as well as the time to onset and percentage of gasping, loss of balance, vocalizations, muscular excitation and gagging. At the end of the exposure, the corneal reflex, rhythmic breathing and sensitivity to pain were each assessed at 10 s intervals for 5 min. Brain activity decreased significantly (P < 0.05) 37.60 s after the start of the exposure to 90% CO(2), which was significantly earlier than in 70N30C, 80N20C and 85N15C exposure, (45.18 s, 46.92 s and 43.27 s, respectively). Before brain activity decreased, all pigs experienced gasping and loss of balance and a 98% muscular excitation. The duration of the muscular excitation was longer in animals exposed to 70N30C, 80N20C and 85N15C than 90C (P < 0.01). After a long exposure time, all animals exposed to 90C died, whereas the 30.4% of animals exposed to N(2)/CO(2) gas mixtures survived. Pigs exposed to 85N15C recovered corneal reflex and sensitivity to pain significantly earlier than when exposed to 90C. Exposure to 90C causes a higher aversive reaction but a quicker loss of consciousness than N(2)/CO(2) gas mixtures. Exposure to N(2)/CO(2) gas mixtures causes a lower percentage of deaths and an earlier recovery of the brain stem activity than 90C, whereas the time to recover the cortical activity is similar. In conclusion, the inhalation of N(2)/CO(2) gas mixtures reduces the aversion compared with high concentrations of CO(2); however, the period of exposure for inducing unconsciousness may be longer in N(2)/CO(2) gas mixtures, and the signs of recovery appear earlier, compared to CO(2).

The welfare of growing pigs in five different production systems: assessment of feeding and housing.

Ninety-one farms were visited over a 2-year period to assess the welfare of growing pigs in five different production systems found either in France or in Spain using the Welfare Quality® protocol. This study focused on animal-based measures as indicators of 'good feeding' and 'good housing'. Multiple Generalized Linear Mixed Models were performed for each measure to evaluate the differences between production systems and to detect possible causal factors. Pigs in the conventional system presented the lowest prevalence of poor body condition, whereas extensive Mallorcan Black pigs and extensive Iberian pigs were associated with a decreased prevalence of bursitis and pig dirtiness. The straw-bedded system presented a lower prevalence of bursitis, but poorer hygiene and more susceptibility of poor body condition than the conventional system. The age of the animals had a significant effect on the appearance of bursitis in the three intensive systems studied. The type of floor was a significant causal factor of bursitis and pig dirtiness in the conventional system and among intensive Iberian pigs. The feeding system was another causal factor of pig dirtiness on more than 50% of the body in the conventional system, whereas pig dirtiness on less than 50% of the body was influenced by the age of the animals. The prevalence of huddling animals in the conventional system was associated with the highest stocking densities and the lowest environmental temperatures. The results indicate that there were important differences between production systems based on animal-based indicators of the good feeding and housing principles. The recording of the age of the animals, type of floor, feeding system, stocking density and environmental temperature can be useful to predict the appearance of a given welfare measure of 'good housing' on a farm.

Stunning pigs with nitrogen and carbon dioxide mixtures: effects on animal welfare and meat quality.

The aim of this study was to assess the effect of exposure to the gas mixtures of 70% nitrogen (N2) and 30% carbon dioxide (CO2; 70N30C), 80% N2 and 20% CO2 (80N20C) and 85% N2 and 15% CO2 (85N15C) on aversion, stunning effectiveness and carcass, as well as meat quality in pigs, and to compare them with the commercial stunning of 90% CO2 (90C). A total of 68 female pigs were divided into four groups and stunned with one of the gas mixtures. During the exposure to the gas, behavioural variables (retreat attempts, escape attempts, gasping, loss of balance, muscular excitation and vocalizations) were recorded, and at the end of the stunning, corneal reflex and rhythmic breathing were assessed. After slaughter, meat quality parameters such as pH at 45 min post mortem (pH45) and at 24 h post mortem (pHu), electrical conductivity, drip loss and colour, in the Longissimus thoracis (LT) and Semimembranosus (SM) muscles were measured, and the presence of ecchymosis on the hams was noted. The PROC MIXED and the PROC GENMOD of SAS® were used to analyse the parametric and binomial variables, respectively. The 'gas mixture' was always considered a fixed effect and the 'live weight' as a covariate. To assess the correlation between meat quality and behaviour measures, PROC CORR was used. Pigs exposed to 90C showed a higher percentage of escape attempts and gasping, a lower percentage of vocalization and shorter muscular excitation phase than pigs exposed to the other N2 and CO2 mixtures (P < 0.05). After stunning, no pig exposed to 90C showed corneal reflex or rhythmic breathing, whereas 85% and 92% of the animals exposed to N2 and CO2 mixtures showed corneal reflex and rhythmic breathing, respectively. Animals stunned with 80N20C and 85N15C had a lower pH45 (P < 0.01) than animals exposed to 90C. Electrical conductivity in the SM muscle was lower (P < 0.001) in 90C and 70N30C pigs than in 80N20C and 85N15C pigs, whereas in LT, it was lower (P < 0.05) in 90C pigs than in 85N15C. As the CO2 concentration of the gas mixture was decreased, the prevalence of exudative pork increased. Twenty-five percent of animals exposed to N2 and CO2 mixtures (n = 68) had ecchymosis in their carcasses, whereas no animal stunned with 90C had ecchymosis. In conclusion, although N2 and CO2 stunning exhibit fewer signs of aversion than 90C, their induction time to unconsciousness is longer, and this may negatively affect meat and carcass quality.