Correction: Mixed management in growing and finishing pigs: Differences between gender and their impacts on behavior, growth performance, and physiological parameters.

[This corrects the article DOI: 10.1371/journal.pone.0284481.].

Changes in cortisol and cortisone in hair of pigs reared under heat stress conditions.

Heat stress accounts for millions of dollars in losses for swine producers worldwide. The aim of the present study was to determine and evaluate cortisol and cortisone in hair as indicators of thermal stress in growing pigs reared under high environmental temperatures. The study was carried out in two independent batches of commercial crosses of Lean Duroc and Pietrain in trials 1 and 2, respectively, during the growing period (from 40 to 100 kg; 81 days in trial 1 and 77 days in trial 2) in the same commercial farm in Spain during the summers of 2020 and 2021. In both cases, four rooms were used. In Trial 1, Room 1 had cooling and 11 pigs per pen; Room 2 had no cooling and 13 pigs per pen; Room 3 had no cooling and 11 pigs per pen, and Room 4 had cooling and 13 pigs per pen. In Trial 2, Rooms 2 and 3 had cooling and rooms 1 and 4 had no cooling, and all of them had 13 pigs per pen. Mean THI value was higher (p < 0.0001) in rooms without cooling systems (75.0 trial 1; 74.9 trial 2) than with them (71.3 trial 1; 71.7 trial 2). A total of four pens per room (16 in total) was selected for analysis of hair corticoids and all pigs inside were sampled at the end of the study. Fifty percent of the pigs were males (castrated and intact in trial 1 and 2, respectively) and 50% females. In total, 44, 52, 44, and 52 pigs, respectively, were sampled in four rooms from the first trial and 52 for each of four rooms in Trial 2. Cortisol concentrations in hair did not show any significant change in relation to cooling-non-cooling in any trial. However, hair cortisone concentration was 172.3 pg./mg and 105.8 pg./mg less (p < 0.001) in pigs housed with cooling systems compared to those without them in Trial 1 and 2, respectively. In addition, the cortisone/cortisol ratio, which is an estimator of the activity of 11ß-hydroxysteroid dehydrogenase (11ß-HSD) type 2, was also greater in rooms without cooling than in rooms with cooling in both trials (p < 0.0001 and p = 0.0105 for Trials 1 and 2, respectively). In relation to the sex effect, the results showed greater levels in females than in castrated males both in cortisone and the cortisol/cortisone ratio while cortisol hair levels were greater in intact males than in females. Therefore, the use of cortisone and the estimation of 11ß-HSD type 2 activity in hair is recommended to evaluate the chronic stress produced by high environmental conditions in pigs instead of using hair cortisol concentrations alone.

Reducing Stocking Densities and Using Cooling Systems for More Adapted Pigs to High Temperatures When Reared in Intensive Conditions.

This study is aimed at evaluating the effect of reducing stocking density and using cooling systems to mitigate the negative effects of high temperatures in growing pigs (females and castrated males) reared in intensive conditions (from 25 to 100 kg) during summer (June to October 2020). The experimental design was a 2 × 2 factorial where pigs were provided with an evaporative cooling system and/or raised at regular or at lower stocking densities (i.e., 0.68 to 0.80 m2/animal). Treatments were distributed in four different rooms containing sex-balanced pens with either castrated males or females. Temperature and humidity were recorded throughout the experiment, and the temperature-humidity index was calculated. Heat stress (HS) on pigs was measured through changes in animals' performance, animal-based indicators (dirtiness and activity budget) and physiological indicators (neutrophil/lymphocyte ratio and hair cortisol). The use of cooling, lowering stocking density and the combination of both strategies had positive effects on pigs' final body weight (+5 kg, +3 kg, +9 kg, respectively; p < 0.001). The prevalence of dirtiness was similar at the stocking densities tested, and no clear effect of the cooling system was found. Both mitigation strategies lowered the physiological indicators of stress, although only hair cortisone can be considered an indicator of HS. In conclusion, both mitigation strategies are effective in improving pig welfare and performance, especially when both are combined. The severity of the stocking density effect may depend on the severity of the temperature.

Copy Number Variation on ABCC2-DNMBP Loci Affects the Diversity and Composition of the Fecal Microbiota in Pigs.

Genetic variation in the pig genome partially modulates the composition of porcine gut microbial communities. Previous studies have been focused on the association between single nucleotide polymorphisms (SNPs) and the gut microbiota, but little is known about the relationship between structural variants and fecal microbial traits. The main goal of this study was to explore the association between porcine genome copy number variants (CNVs) and the diversity and composition of pig fecal microbiota. For this purpose, we used whole-genome sequencing data to undertake a comprehensive identification of CNVs followed by a genome-wide association analysis between the estimated CNV status and the fecal bacterial diversity in a commercial Duroc pig population. A CNV predicted as gain (DUP) partially harboring ABCC2-DNMBP loci was associated with richness (P = 5.41 × 10-5, false discovery rate [FDR] = 0.022) and Shannon α-diversity (P = 1.42 × 10-4, FDR = 0.057). The in silico predicted gain of copies was validated by real-time quantitative PCR (qPCR), and its segregation, and positive association with the richness and Shannon α-diversity of the porcine fecal bacterial ecosystem was confirmed in an unrelated F1 (Duroc × Iberian) cross. Our results advise the relevance of considering the role of host-genome structural variants as potential modulators of microbial ecosystems and suggest the ABCC2-DNMBP CNV as a host-genetic factor for the modulation of the diversity and composition of the fecal microbiota in pigs. IMPORTANCE A better understanding of the environmental and host factors modulating gut microbiomes is a topic of greatest interest. Recent evidence suggests that genetic variation in the pig genome partially controls the composition of porcine gut microbiota. However, since previous studies have been focused on the association between single nucleotide polymorphisms and the fecal microbiota, little is known about the relationship between other sources of genetic variation, like the structural variants and microbial traits. Here, we identified, experimentally validated, and replicated in an independent population a positive link between the gain of copies of ABCC2-DNMBP loci and the diversity and composition of pig fecal microbiota. Our results advise the relevance of considering the role of host-genome structural variants as putative modulators of microbial ecosystems and open the possibility of implementing novel holobiont-based management strategies in breeding programs for the simultaneous improvement of microbial traits and host performance.

Mixed management in growing and finishing pigs: Differences between gender and their impacts on behavior, growth performance, and physiological parameters.

Mixing, a common management strategy used to regroup pigs, has been reported to impair individual performance and affect pig welfare because of the establishment of a new social hierarchy after regrouping. In this study we aimed to determine whether mixing management (non-mixed vs. mixed) and gender (gilts vs. barrows) affect the social and non-social behavior, performance, and physiological parameters of pigs. A total of 96 growing pigs (48 barrows and 48 females) were separated into two treatments: control (CT)-pigs that were mixed once during the growing-finishing period; and social stress (SS)-pigs that were mixed thrice during the growing-finishing period. We recorded social and non-social behaviors, injury score, performance, and physiological parameters during the experimental period. Data were grouped by the period, based on each mix performed, and overall values. The statistical analysis performed considered gender and treatment. For treatment, during period-II and III, the SS group presented the highest frequency of agonistic interactions (AI), stayed longer lying laterally (LL) and sternly (LS), and explored more enrichment material (ER) than the CT group. Furthermore, SS pigs presented the highest injury score in the ear, head, and middle and posterior regions. Compared to the females, the barrows spent more time at the electronic feed station and initiated most of the agonistic interactions during period-II, and they presented a higher injury score for the ear and head regions during period-III. In conclusion, repeated regrouping significantly affected social and feeding behavior without severely altering performance and physiological parameters. Furthermore, different patterns of social and feeding behavior, agonistic interactions, and injury scores between barrows and females were observed. This study provides an understanding of the impact of mixing management and gender differences on pigs, and this knowledge can be used to improve swine productivity and welfare.

Hierarchy Establishment in Growing Finishing Pigs: Impacts on Behavior, Growth Performance, and Physiological Parameters.

In recent years, an increased number of studies have dealt with the analysis of social dominance related to animal behavior, physiology, and performance. This study aimed to investigate whether hierarchical ranking affects the coping style, non-social behavior during open field and novel object tests, performance, and physiological parameters of pigs. A total of 48 growing pigs (24 barrows and 24 females) were mixed three times during the growing-finishing period. The social and non-social behaviors of pigs were directly noted, and three behavioral tests were performed during the experimental period. Performance and physiological parameters were also recorded. Statistical analysis considered hierarchical classification (dominant vs. intermediary vs. subordinate) and p-values ≤ 0.05 were considered significant. After three regroupings, the pigs in different hierarchical classifications showed no change in hair cortisol values and open-field and novel object tests. Mean corpuscular hemoglobin concentration and leukocyte values increased in intermediary pigs, and the lowest counts were found in pigs classified as dominants. Furthermore, dominant pigs visited the feeder more but spent shorter time there compared to subordinate and intermediary pigs. Our results suggest that hierarchical classification influenced feeding behavior and physiological parameters without affecting cortisol values and growth performance, demonstrating a possible compensation skill.

Should Animal Welfare be Included in Educational Programs? Attitudes of Secondary and University Students from Eight EU Countries.

Animal Welfare (AW) educational programs aim to promote positive attitudes of future generations toward animal production systems. This study investigated whether secondary and university students in the majors that are not related to AW teaching believe that this concept should be included also in their educational programs. The determinant factors affecting students' attitudes toward such a decision were analyzed. This research has focused on eight European countries (Spain, the United Kingdom, Poland, Greece, Lithuania, Romania, Italy, and Sweden) targeting 3,881 respondents composed of 1,952 secondary and 1,929 university students. The results showed that female university students with a high level of subjective and objective knowledge on AW and who required more restrictive AW regulations, gave support to include the concept in their educational programs. However, Students who support medical experiments that use animals to improve human health were less likely to accept AW education. Furthermore, students in Italy compared to those in Sweden were prone to support AW educational programs. Results highlight the importance of teaching the AW concept as a comprehensive teaching tool at universities and schools' programs as it may constitute a starting point for a more sustainable society toward improving animal living conditions, mainly in the Mediterranean countries in secondary schools.

Induced Stress and Tactile Stimulation Applied to Primiparous does and their Consequences on Maternal Behavior, Human-Animal Relationships, and Future Offspring's Sexual Disorders.

The aim of this study was to investigate whether tactile stimulation in rabbits during the gestation phase improve the maternal behavior and human-animal relationships as well as the effects on reproductive behavior of male kits when reached maturity compared to induced stress. A total of 33 primiparous New Zealand does were selected after pregnancy confirmation and allocated in a randomized complete block design. The treatments applied were as follows: (C) animals not stimulated during the experimental period; (TS) animals that received tactile stimulation; and (SS) does which were immobilized. The nest building behavior as well as the weight, sexual behavior, mortality, and semen analysis of the offspring was recorded. In addition, the novel object, flight distance, social isolation, and human-approach tests were conducted. Under the conditions of the present trial, TS animals showed more trust in the unfamiliar observer when compared to the other two treatments. The treatments applied to the females (TS and SS) were sufficient to confirm that the control group presented better values for the number of stillbirths and the proportion of deaths in the first week. Finally, the handling of does reduce the males' ejaculation and sperm presence but not inhibited sexual behavior or impaired semen quality. It is possible to conclude that TS did not impair does welfare or maternal behavior and it improved the human-animal relationship, however there was a negative impact on the litter. More studies that directly assess impact on the future reproductive capacity of the offspring are necessary.

Consumers' Expectations about Meat from Surgical Castrated or Immunocastrated Male and Female Iberian Pigs.

A common practice in Iberian pigs is the castration of both males and females, and it can be carried out surgically or by immunization against gonadotropin-releasing factor (GnRF). The aim of this work was to determine consumers' overall liking and expectations towards Iberian pork from five different sex types (castrated females, entire females, GnRF-vaccinated females, castrated males and GnRF-vaccinated males), as well as to know the attitudes and beliefs of consumers towards castration and immunocastration. Loins from 83 Iberian pigs were collected and evaluated by 252 consumers in Barcelona and Madrid. Consumers evaluated the five types of meat in three situations: blind condition (tasting the product), expectations (without tasting) and informed condition. Finally, attitudes and beliefs towards castration and immunocastration were also determined. Results distinguished three segments of consumers labeled as "Indifferent", "Against castration and immunocastration" and "Against immunocastration". Meat from castrated males had higher overall liking scores in the blind condition. Expectations towards pork depending on its sex type affect consumer-informed acceptability; thus, it is important to consider marketing strategies to avoid or direct the effect of the information provided on the acceptability of the pork.

Leveraging host-genetics and gut microbiota to determine immunocompetence in pigs.

BACKGROUND: The gut microbiota influences host performance playing a relevant role in homeostasis and function of the immune system. The aim of the present work was to identify microbial signatures linked to immunity traits and to characterize the contribution of host-genome and gut microbiota to the immunocompetence in healthy pigs. RESULTS: To achieve this goal, we undertook a combination of network, mixed model and microbial-wide association studies (MWAS) for 21 immunity traits and the relative abundance of gut bacterial communities in 389 pigs genotyped for 70K SNPs. The heritability (h2; proportion of phenotypic variance explained by the host genetics) and microbiability (m2; proportion of variance explained by the microbial composition) showed similar values for most of the analyzed immunity traits, except for both IgM and IgG in plasma that was dominated by the host genetics, and the haptoglobin in serum which was the trait with larger m2 (0.275) compared to h2 (0.138). Results from the MWAS suggested a polymicrobial nature of the immunocompetence in pigs and revealed associations between pigs gut microbiota composition and 15 of the analyzed traits. The lymphocytes phagocytic capacity (quantified as mean fluorescence) and the total number of monocytes in blood were the traits associated with the largest number of taxa (6 taxa). Among the associations identified by MWAS, 30% were confirmed by an information theory network approach. The strongest confirmed associations were between Fibrobacter and phagocytic capacity of lymphocytes (r = 0.37), followed by correlations between Streptococcus and the percentage of phagocytic lymphocytes (r = -0.34) and between Megasphaera and serum concentration of haptoglobin (r = 0.26). In the interaction network, Streptococcus and percentage of phagocytic lymphocytes were the keystone bacterial and immune-trait, respectively. CONCLUSIONS: Overall, our findings reveal an important connection between gut microbiota composition and immunity traits in pigs, and highlight the need to consider both sources of information, host genome and microbial levels, to accurately characterize immunocompetence in pigs.

Intramuscular vs. Intradermic Needle-Free Vaccination in Piglets: Relevance for Animal Welfare Based on an Aversion Learning Test and Vocalizations.

The aim of the present study was to compare intramuscular injection with a needle and intradermic needle-free vaccinations against porcine reproductive and respiratory syndrome (PRRS) in piglets at 28 days old by studying behavioral and physiological reactions. A total of 72 piglets divided into 2 sex-balanced batches were assessed. Within each batch, the piglets were divided into three treatments, which were Hipradermic (0.2 ml of UNISTRAIN® PRRS vaccine administered with an intradermic needle-free device), Intramuscular (IM, 2.0 ml of vaccine), and Control (not vaccinated). Before the vaccination, the piglets were trained to cross a 4-m-long raceway to perform an aversion learning test. The day of vaccination, the time taken to cross the raceway was registered for each piglet at different times: prior to the vaccination and 10 min, 2, 24, 48, and 72 h after the vaccination, to measure variations in these times as signs of aversion to the vaccination process. Vocalizations, as potential signs of pain, were recorded as well at the end of this raceway to analyze their frequency (Hz), duration, and level of pressure (dB) at the moment of vaccination. Salivary cortisol, as a sign of the HPA-axis activity, was assessed 10 min after the vaccination. In addition, activity budgets, local reaction to the vaccine, and serological titer were also considered in the study. Ten minutes after the vaccination, the IM piglets took longer (p < 0.001) to cross the raceway than did the Hipradermic and Control piglets. Vocalizations were significantly different between the three treatments: the Control piglets produced vocalizations with the lowest frequency (p < 0.001) and level of pressure (p < 0.001), and IM with the highest, with Hipradermic in a significant intermediate position (p < 0.001). Accordingly, the day of the vaccination, IM and Hipradermic animals were lying on the side of the vaccine administration a greater proportion of time than were the Control piglets (10, 11, and 6%, respectively; p = 0.027). Salivary cortisol was not significantly different between treatments. The serum titer of antibodies against the PRRS was higher (p < 0.001) in both vaccinated treatments in comparison to the Control piglets. It is concluded that the Hipradermic needle-free vaccination may result in a less aversive experience in piglets than did intramuscular vaccination.

A gene co-association network regulating gut microbial communities in a Duroc pig population.

BACKGROUND: Analyses of gut microbiome composition in livestock species have shown its potential to contribute to the regulation of complex phenotypes. However, little is known about the host genetic control over the gut microbial communities. In pigs, previous studies are based on classical "single-gene-single-trait" approaches and have evaluated the role of host genome controlling gut prokaryote and eukaryote communities separately. RESULTS: In order to determine the ability of the host genome to control the diversity and composition of microbial communities in healthy pigs, we undertook genome-wide association studies (GWAS) for 39 microbial phenotypes that included 2 diversity indexes, and the relative abundance of 31 bacterial and six commensal protist genera in 390 pigs genotyped for 70 K SNPs. The GWAS results were processed through a 3-step analytical pipeline comprised of (1) association weight matrix; (2) regulatory impact factor; and (3) partial correlation and information theory. The inferred gene regulatory network comprised 3561 genes (within a 5 kb distance from a relevant SNP-P < 0.05) and 738,913 connections (SNP-to-SNP co-associations). Our findings highlight the complexity and polygenic nature of the pig gut microbial ecosystem. Prominent within the network were 5 regulators, PRDM15, STAT1, ssc-mir-371, SOX9 and RUNX2 which gathered 942, 607, 588, 284 and 273 connections, respectively. PRDM15 modulates the transcription of upstream regulators of WNT and MAPK-ERK signaling to safeguard naive pluripotency and regulates the production of Th1- and Th2-type immune response. The signal transducer STAT1 has long been associated with immune processes and was recently identified as a potential regulator of vaccine response to porcine reproductive and respiratory syndrome. The list of regulators was enriched for immune-related pathways, and the list of predicted targets includes candidate genes previously reported as associated with microbiota profile in pigs, mice and human, such as SLIT3, SLC39A8, NOS1, IL1R2, DAB1, TOX3, SPP1, THSD7B, ELF2, PIANP, A2ML1, and IFNAR1. Moreover, we show the existence of host-genetic variants jointly associated with the relative abundance of butyrate producer bacteria and host performance. CONCLUSIONS: Taken together, our results identified regulators, candidate genes, and mechanisms linked with microbiome modulation by the host. They further highlight the value of the proposed analytical pipeline to exploit pleiotropy and the crosstalk between bacteria and protists as significant contributors to host-microbiome interactions and identify genetic markers and candidate genes that can be incorporated in breeding program to improve host-performance and microbial traits. Video Abstract.

Sex Differences in Group Composition and Habitat Use of Iberian Free-Range Pigs.

The aim of the present work was to study group size, group composition and habitat use of Iberian pigs along the year when reared outdoor. This consists of a regimen in which animals are reared free range from 2 months of age until at least 14 months of age. In a first stage, animals are supplemented with concentrates, and in a second, called montanera, pigs eat just natural resources in areas with no more than two pigs per hectare. In these systems, males are castrated to avoid boar taint and females spayed to avoid the attraction and mounting by wild boars. The study was carried out in five different farms allocated in the south-west of Spain during 2 consecutive years, from March 2012 to February 2014, under the montanera regimen, and with a total of 995 animals observed (498 males and 497 females). The data were analyzed with SAS by means of general models and proc mixed. Mean group size along the year was of 17 ± 12.9 individuals, but this was significantly lower (P < 0.05) during the montanera (12 ± 0.8) and at midday (13 ± 0.8). Groups were bigger (P < 0.05) when they were more than 50 m from a tree (23 ± 1.8), or <10 m from the shelter (25 ± 1.5), the feeding area (31 ± 3.1) and the water-bath area (25 ± 1.5). Nine percent of the groups were solitary animals, being higher (P = 0.0286) during the montanera (11%) than the rest of the year (8%) and being formed in 68% by males. Males were less involved in mixed groups than were females (75% vs. 91%), especially in spring, where the largest (P < 0.0001) male groups were found. Female groups were less frequent and smaller (P < 0.0001) than were male and mixed groups. In conclusion, although males were castrated at a very young age, they showed a different behavior than females, forming in bachelor groups during the spring and being less involved in mixed groups and with more solitary animals. During the montanera, when animals were feeding on acorns and other natural resources, groups were smaller and closer to the trees, solitary males reaching a maximum percent.

To Provide a Double Feeder in Growing Pigs Housed under High Environmental Temperatures Reduces Social Interactions but Does Not Improve Weight Gains.

Heat stress and competition for food are two major challenges in pigs reared in intensive conditions. The aim of the present work was to study the effect of providing a double feeder for pigs reared under two different environmental temperatures. In addition, two types of flooring, of 100% slat and 30% slat 70% concrete, were also considered. A total of 256 pigs in the growing-finishing period (from 27 kg to 110 kg) were housed using two environmental temperatures: control (from 18 °C to 25 °C) and heat stress (above 30 °C six hours a day). They were housed in 32 pens of 8 pigs each, distributed into 4 rooms (16 with one feeder and 16 with two). Pigs subjected to temperatures above 30 °C up to six hours had lower body weight gains than pigs subjected to a maximum temperature of 25 °C, confirming that thermal stress negatively affects performance in pigs. In addition, heat stress affected the final product by decreasing the lean percentage of carcasses by 2.6%. A double feeder reduced the presence of negative social behavior, especially in the feeding area, but body weight was lower than when one single feeder was used. A 30% slat 70% concrete floor showed better results in the pig stress indicators and body weights than 100% slat. It is concluded that providing a double feeder in the pens, although reducing the presence of negative social interactions, negatively affected body weight, in comparison to pigs fed with just one feeder.

Animal Welfare Assessment Protocol for Does, Bucks, and Kit Rabbits Reared for Production.

Rabbits are the third species in terms of number of animals reared for meat production in the world. However, in comparison to other species, very few studies have focused on their welfare. The aim of the present study is to implement an animal welfare-assessment protocol developed through a multidimensional approach and containing a number of animal-based measures for bucks, does, and kit rabbits. Thirty Spanish farms with conventional cages in the first year of integration in an animal welfare certification scheme were visited during 2019 and audited by the same auditor. The protocol is divided into four principles and 11 criteria. The Good Feeding principle includes eight parameters (one animal-based), Good Housing includes 15 parameters (six animal-based), Good Health includes 26 parameters (16 animal-based), and Appropriate Behavior contains nine parameters (four animal-based). In general, the main problems found were the absence of platforms, low space allowance and low height of the cage, inappropriate systems for performing emergency killing, insufficient protection of does from other adjacent does when housed individually, and absence of enrichment material. To a minor degree, but also found in an important number of farms, was a lack of temperature data records, high replacement rates of does, and lack of mortality-rate data records. However, in general, most of the farms obtained a good overall score, the maximum found being 73 out of 100 points. Nevertheless, none of the farms reached an excellent score, and four farms were scored below the 55 points required in the animal welfare certification scheme. The Good Feeding principle obtained the highest score, reaching excellent in all farms, and Appropriate Behavior the lowest one, with values ranging from 21 to 41 points out of 100. The results probably show how, for years, rabbit producers have been very focused on feeding needs and very little attention has been paid to behavioral needs.

Testing an Animal Welfare Assessment Protocol for Growing-Rabbits Reared for Meat Production Based on the Welfare Quality Approach.

The objective of the present study is to present an animal welfare assessment protocol for growing-rabbits for discussion after its implementation in 32 farms from Spain and Portugal. The protocol comprises the principles of Good Feeding, Good Housing, Good Health and Appropriate Behaviour of the Welfare Quality protocols and includes 36 welfare parameters. Overall, the protocol showed a good capacity for discrimination between farms, with scores ranging 44 to 82 points. The protocol seems reliable for the assessment of animal welfare after proper training of auditors. However, for the criteria social behaviour and other behaviours, further research is needed to ascertain if the methodology and times of observation used are appropriate. Some farms had high mortality rates with a low prevalence of health problems, while others had low mortality rates with high prevalence of health problems due to different managements of culling. The protocol should be improved, to impede farms with high mortality rates but a low prevalence of health issues the day of the audit from obtaining better scores than the second type of farms, by limiting the compensation in key measures. The main points to be solved in the growing-rabbit farms were: to provide more space to the animals; register the number of animals culled accurately; change cervical dislocation for another killing method and provide the farmers training in animal welfare.

Environmental and Management Factors Affecting the Time Budgets of Free-Ranging Iberian Pigs Reared in Spain.

Understanding the natural behaviour of pigs in free-range conditions facilitates interpretation of their behaviour in intensive conditions. Studying six different farms over two years at different seasons, with climatic and management variations, allowed for a general description of Iberian pig behaviour and which factors have an influence on it. The main activity found was resting (56.5% of the time observed), followed by exploratory behaviour. However, this exploratory behaviour was higher when animals were fed only with natural resources than when fed with concentrates (50% versus 17.8%, respectively). In addition, pigs used bathing areas in summer that were not visited in winter. Negative social behaviour was seen more frequently than positive social behaviour, accounting, in total, for 1% of the total activity of animals. Pigs situated at the centre of the groups tended to remain more relaxed, while the peripheral animals remained more alert and vigilant. Our results indicate that foraging behaviour accounts for a significant proportion of pigs' active time, but this proportion is much more reduced when pigs are fed concentrates. Therefore, behavioural needs in pigs reared in intensive conditions should consider that exploratory behaviour is reduced when pigs are fed with concentrates.

Gut eukaryotic communities in pigs: diversity, composition and host genetics contribution.

BACKGROUND: The pig gut microbiome harbors thousands of species of archaea, bacteria, viruses and eukaryotes such as protists and fungi. However, since the majority of published studies have been focused on prokaryotes, little is known about the diversity, host-genetic control, and contributions to host performance of the gut eukaryotic counterparts. Here we report the first study that aims at characterizing the diversity and composition of gut commensal eukaryotes in pigs, exploring their putative control by host genetics, and analyzing their association with piglets body weight. RESULTS: Fungi and protists from the faeces of 514 healthy Duroc pigs of two sexes and two different ages were characterized by 18S and ITS ribosomal RNA gene sequencing. The pig gut mycobiota was dominated by yeasts, with a high prevalence and abundance of Kazachstania spp. Regarding protists, representatives of four genera (Blastocystis, Neobalantidium, Tetratrichomonas and Trichomitus) were predominant in more than the 80% of the pigs. Heritabilities for the diversity and abundance of gut eukaryotic communities were estimated with the subset of 60d aged piglets (N = 390). The heritabilities of α-diversity and of the abundance of fungal and protists genera were low, ranging from 0.15 to 0.28. A genome wide association study reported genetic variants related to the fungal α-diversity and to the abundance of Blastocystis spp. Annotated candidate genes were mainly associated with immunity, gut homeostasis and metabolic processes. Additionally, we explored the association of gut commensal eukaryotes with piglet body weight. Our results pointed to a positive contribution of fungi from the Kazachstania genus, while protists displayed both positive (Blastocystis and Entamoeba) and negative (Trichomitus) associations with piglet body weight. CONCLUSIONS: Our results point towards a minor and taxa specific genetic control over the diversity and composition of the pig gut eukaryotic communities. Moreover, we provide evidences of the associations between piglets' body weight after weaning and members from the gut fungal and protist eukaryote community. Overall, this study highlights the relevance of considering, along with that of bacteria, the contribution of the gut eukaryote communities to better understand host-microbiome association and their role on pig performance, welfare and health.

Should Animal Welfare Regulations Be More Restrictive? A Case Study in Eight European Union Countries.

Increasingly, intensive livestock production systems have increased societal concern regarding the current animal welfare standards. We investigated whether individuals in their roles as consumers and citizens believe that the current European regulations regarding animal welfare should be more restrictive. Factors affecting this decision were assessed by analyzing respondents' understanding of animal welfare-related issues, their subjective and objective knowledge levels, the credibility they assign to different information sources, their perceptions toward the current restrictiveness of animal welfare standards, and their socioeconomic characteristics. Data were collected using a semi-structured questionnaire distributed in eight European Union (EU) countries (Spain, the United Kingdom, Poland, Greece, Lithuania, Romania, Italy, and Sweden) with 3860 total responses. The results showed that consumers are more reluctant to adopt more restrictive regulations than respondents in the role of citizens. Respondents from northern European countries (Poland and Sweden) are more likely to support regulations that are more restrictive than the current minimum requirements than respondents from southern countries (Spain and Italy). Women were found to be more concerned with the welfare of pigs and laying hens-lending credibility to the Internet as an information source-and were more likely to support more restrictive animal welfare legislation.

Does the location of enrichment material affect behavior and dirtiness in growing female pigs?

The objective of this study was to investigate if female growing pigs spend similar amounts of time exploring a wooden stick as they do exploring enrichment material regardless of its proximity to the feeder. Forty-eight pigs aged 18 to 26 weeks allocated to 16 pens with three pigs per pen were studied. Fifty percent of the pens had a wooden stick beside the feeder, and the rest had a similar stick opposite to the feeder. Two observers assessed the pigs by means of scan and focal sampling. The pigs spent more time (p < .0001) exploring the wood during the first week than during the rest of the study (10.9% vs 3.6%). The pigs with the wood close to the feeder spent less (p = .0001) time resting (29.9%) and more (p < .0001) time exploring (6.3%) the wood than did pigs with the wood opposite to the feeder (32.4% and 2.5%, respectively). In conclusion, a wooden stick placed close to the feeder was associated with more exploratory behaviorcompared with a similar stick placed opposite to the feeder.