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 ofKazachstaniaspp. 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 ofBlastocystisspp. 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 theKazachstaniagenus, while protists displayed both positive (BlastocystisandEntamoeba) 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.

The molecular basis of compensatory growth in monogastric animals has not yet been fully explored. Herewith, in this study we aim to determine changes in the pig skeletal muscle transcriptome profile during compensatory growth following a feed restriction period. A RNA-Seq experiment was performed with a total of 24 females belonging to a Duroc commercial line. Half of the animals received either a restricted (RE) or ad libitum (AL) diet during the first fattening period (60–125 d of age). After that, all gilts were fed ad libitum for a further ~30 d until the age of ~155 d, when animals were slaughtered and samples of gluteus medius muscle were harvested to perform RNA-Seq analyses and intramuscular fat content determination. During the period following food restriction, RE animals re-fed ad libitum displayed compensatory growth, showed better feed conversion rate and tended to deposit more subcutaneous fat than AL fed animals. Animals were slaughtered in the phase of accelerated growth, when RE animals had not completely compensated the performance of AL group, showing lower live and carcass weights. At intramuscular level, RE gilts showed a higher content of polyunsaturated fatty acids during the compensatory growth phase. The comparison of RE and AL expression profiles allowed the identification of 86 (ǀlog₂Fold-Changeǀ > 1, p_adj < 0.05) differentially expressed (DE) genes. A functional categorization of these DE genes identified AMPK Signaling as the most significantly enriched canonical pathway. This kinase plays a key role in the maintenance of energy homeostasis as well as in the activation of autophagy. Among the DE genes identified as components of AMPK Signaling pathway, five out of six genes were downregulated in RE pigs. Animals re-fed after a restriction period exhibited a less oxidative metabolic profile and catabolic processes in muscle than animals fed ad libitum. The downregulation of autophagy observed in the skeletal muscle of pigs undergoing compensatory growth may constitute a mechanism to increase muscle mass thus ensuring an accelerated growth rate. These results reveal that the downregulation of AMPK Signaling plays an important role in compensatory growth in pigs.

Stress significantly affects the health, welfare, and productivity of farm animals. We performed a longitudinal study to evaluate stress’s effects on pig performance, feeding behavior, and fecal microbiota composition. This study involved 64 Duroc pigs during the fattening period, divided into two experimental groups: a stress group (n = 32) and a control group (n = 32). Stressed groups had less space and were mixed twice during the experiment. We monitored body weight, feed efficiency, feeding behavior, and fecal microbiota composition. Compared to the control group, the stressed pigs exhibited reduced body weight, feed efficiency, fewer feeder visits, and longer meal durations. In the fecal microbiota, resilience was observed, with greater differences between groups when sampling was closer to the stressful stimulus. Stressed pigs showed an increase in opportunistic bacteria, such as Streptococcus, Treponema and members of the Erysipelotrichaceae family, while control pigs had more butyrate- and propionate-producing genera like Anaerobutyricum, Coprococcus and HUN007. Our findings confirm that prolonged stress negatively impacts porcine welfare, behavior, and performance, and alters their gut microbiota. Specific microorganisms identified could serve as non-invasive biomarkers for stress, potentially informing both animal welfare and similar gut-brain axis mechanisms relevant to human research.

Breeding animals to produce more robust and disease-resistant pig populations becomes a complementary strategy to the more conventional methods of biosecurity and vaccination. The objective of this study was to explore the ability of a panel of genetic markers and immunity parameters to predict the survival rates during a natural PRRSV outbreak. Ten-week-old female Duroc pigs (n = 129), obtained from 61 sows and 20 boars, were naturally infected with a highly pathogenic PRRSV genotype 1 strain. Prior to infection, piglets were screened for immunity parameters (IgG levels in plasma and SOX13 mRNA expression in blood) and genetic markers previously associated to PRRSV immune response and immunity traits. Additionally, the 20 boars were genotyped with a panel of 132 single nucleotide polymorphisms (SNPs). Survival analysis showed that mortality was significantly higher for animals with low basal IgG levels in plasma and/or high SOX13 mRNA expression in blood. The genotypes of sires for SNPs associated with IgG plasma levels, CRP in serum, percentage of γδ T cells, lymphocyte phagocytic capacity, total number of lymphocytes and leukocytes, and MCV and MCH were significantly associated with the number of surviving offspring. Furthermore, CD163 and GBP5 markers were also associated to piglet survival. The effects of these SNPs were polygenic and cumulative, survival decreased from 94 to 21% as more susceptible alleles were accumulated for the different markers. Our results confirmed the existence of genetic variability in survival after PRRSV infection and provided a set of genetic markers and immunity traits associated with PRRS resistance.

Organic livestock farming is committed to high environmental and animal welfare standards, although pathologies such as post-weaning diarrhoea (PWD) may appear. The main objective of this study was to assess nutritional strategies to prevent PWD in organic piglets. A total of 134 weaned piglets were fed one of three diets: high crude protein (17.8%, HCP), low crude protein (16.8%, LCP), and low crude protein supplemented with liquid whey (LCP+W). Piglets were assessed weekly for four weeks on the following parameters: diarrhoea incidence, additional health parameters, average daily gain, and behaviour. Faecal samples were taken to analyse the intestinal microbiota composition. Data were analysed using LMM and GLMM models and Shannon and Whittaker indexes. No significant effect of diet on diarrhoea incidence was found, but the LCP+W diet increased average daily gain. Pigs fed the LCP+W diet presented a lower percentage of drinking and negative social behaviour compared with the HCP diet, and LCP pigs presented higher exploration compared with HCP. In addition, LCP+W piglets showed a higher abundance of the beneficial genus Frisingicoccus. Although liquid whey did not reduce diarrhoea incidence, the benefits found in growth, microbiota composition, and reduced negative social behaviour indicate that it could be an optimal supplement to organic diets.

Background: The modulation, activation, and differentiation of several immune cells is highly dependent on lipid metabolism. The objective of this study was to analyse the genetic determinism of the porcine plasma lipidome and its association to the animal immune capacity and production performance. The analysis of the blood lipidome of 300 60-day-old Duroc pigs allowed semi-quantification of 982 circulating lipid molecules. We evaluated the genetic determinism of the lipidome abundances, as well as their phenotypic and genetic correlations with health, stress, and carcass phenotypes. Results: Triacylglycerols were the most abundant lipid class among the plasma lipid features, followed by glycerophosphocholines, glycerophosphoethanolamines, diacylglycerols, and fatty acids/esters. Lipidome abundances showed low to moderate phenotypic correlations with the health and production traits, which clustered in two groups with opposite phenotypic correlation patterns with the lipidome. Mean heritability estimates for the circulating lipids abundance was generally low, but 184 lipid molecules showed significant heritability ranging between 0.25 and 0.85. At the genetic level, the percentage and phagocytic capacity of lymphocytes, the proportion of γδ T lymphocytes, and the cortisol concentration in hair were especially correlated with the lipidome, showing more than 200 significant genetic correlations with different lipidic compounds. Putative identification of associated metabolites by mass similarity revealed a large presence of phospholipids and glycerolipids among lipid molecules genetically correlated with immunity traits. Regarding production traits, fatness and lean meat measures showed an opposite pattern of genetic correlations with the porcine lipidome. Lipids positively correlated with fatness were mainly composed of diacyl- and triacyl-glycerides, while potential ceramides and phospholipids were more abundant among the lipids positively correlated with lean meat content at the genetic level. Conclusions: Our results demonstrate a genetic determinism of the porcine blood lipidomic profile and suggest genetic correlations of the lipidome abundances with health and production performance phenotypes. We identify potential lipid biomarkers for assessing animal health and productivity.