Low Residual Feed Intake Pigs Research Articles

Hygiene of housing conditions and proinflammatory signals alter gene expressions in porcine adipose tissues and blood cells.

Adipose tissue is an organ with metabolic, endocrine and immune functions. In this tissue, the expressions of genes associated with several metabolic pathways, including lipid metabolism, have been shown to be affected by genetic selection for feed efficiency, an important trait to consider in livestock. We hypothesized that the stimulation of immune system caused by poor hygiene conditions of housing impacts the molecular and cellular features of adipose tissue and that the impact may differ between pigs that diverge in feed efficiency. At the age of 12 weeks, Large White pigs from two genetic lines divergent for residual feed intake (RFI) were housed in two contrasting hygiene conditions (good vs poor). After six weeks of exposure, pigs were slaughtered (n=36). Samples of blood, subcutaneous (SCAT) and perirenal (PRAT) adipose tissues were collected for cell response and gene expression investigations. The decrease in the relative weight of PRAT was associated with a decline in mRNA levels of FASN, ME, LCN2 and TLR4 (P<0.05) in pigs housed in poor conditions compared with pigs housed in good conditions for both RFI lines. In SCAT, the expressions of only two key genes (PPARG and TLR4) were significantly affected by the hygiene of housing conditions. Besides, the mRNA levels of both LCN2 and GPX3 were influenced by the RFI line (P<0.05). Because we suspected an effect of poor hygiene at the cellular levels, we investigated the differentiation of stromal vascular cells isolated from SCAT in vitro in the absence or presence of a pro-inflammatory cytokine, Tumor Necrosis Factor-α (TNF-α). The ability of these cells to differentiate in the absence or presence of TNF-α did not differ among the four groups of animals (P>0.05). We also investigated the expressions of genes involved in the immune response and lipid metabolism in whole blood cells cultured in the absence and presence of LPS. The hygiene conditions had no effect but, the relative expression of the GPX3 gene was higher (P<0.001) in high RFI than in low RFI pigs while the expressions of IL-10 (P=0.027), TGFβ1 (P=0.023) and ADIPOR2 (P=0.05) genes were lower in high RFI than in low RFI pigs. Overall, the current study indicates that the hygiene of housing had similar effects on both RFI lines on the expression of genes in adipose tissues and on the features of SCAT adipose cells and whole blood cells in response to TNF-α and LPS. It further demonstrates that the number of genes with expression impacted by housing conditions was higher in PRAT than in SCAT. It suggests a depot-specific response of adipose tissue to the current challenge.

Poor hygiene of housing conditions influences energy metabolism in a muscle type-dependent manner in growing pigs differing in feed efficiency

The ability of pigs to cope with inflammatory challenges may by modified by selection for residual feed intake (RFI), a measure of feed efficiency. In the current study, we evaluated skeletal muscle metabolic responses to degraded hygiene conditions in pigs divergently selected for RFI. At 82 d of age, low RFI and high RFI pigs were housed in either poor or good hygiene conditions. After a 6-week challenge, the poor hygiene conditions induced a decrease in growth performance (P < 0.001) and in plasma IGF-I concentrations (P < 0.003) in both lines. In the slow-twitch oxidative semispinalis muscle, poor hygiene conditions induced a shift towards a more oxidative metabolism and an activation of the AMPK pathway in pigs of both RFI lines. In the fast-twitch glycolytic longississimus muscle, poor hygiene conditions were associated to a less glycolytic metabolism in the HRFI line only. Poor hygiene conditions also increased the protein level of lipidation of microtubule-associated protein 1 light-chain 3β (LC3-II) in both RFI lines, suggesting an activation of the autophagy pathway. Altogether, the data revealed muscle-type specific metabolic adaptations to poor hygiene conditions, which may be related to different strategies to fuel the activated immune system.

Effect of moderate forced physical activity on behaviour, lameness and osteochondrosis in growing pigs from two divergent lines selected for feed efficiency

In pig farming, physical constraints and genetic selection for high production are risk factors for the development of leg disorders, such as degraded locomotor activity. Interactions between both factors need to be explored. The study was carried out on two replicates of 80 pure-bred Large White growing-finishing pigs from the 8th generation of two divergent lines selected for low and high residual feed intake (LRFI, HRFI). Each replicate included 40 LRFI pigs and 40 HRFI pigs, housed on partly slatted flooring in a room equipped with a sorter allowing access to electronic self-feeders during two replicates. Ear tags determined the side of the room to which the pigs were oriented after the sorter exit and the distance back to the sorter (short: spontaneous activity, long: forced activity (FA)). Lameness was assessed individually weekly using visual gait scoring. At slaughter (weight of 100 kg), postmortem quantification of osteochondrosis (OC) lesions was performed on both the proximal and distal extremities of the humerus and femur. Low RFI pigs showed a lower feed conversion ratio (P < 0.001). They also showed lower individual numbers of sorter crossings per day and a lower proportion of standing pigs, which confirmed their lower physical activity. Forced activity clearly increased the number of sorter crossings/d/pig (P < 0.001), and the magnitude of the effect of FA was clearly lower in LRFI pigs than in HRFI pigs. The occurrence of gait was low (less than 9% of recorded scores). The proportion of scores classified as stiffness was higher for LRFI pigs than in HRFI pigs (P < 0.0001). The average lameness score was also higher for LRFI pigs and lower with FA (P < 0.05). The pigs of the LRFI line showed higher OC scores on both the proximal humerus and femur (P < 0.001) and lower OC scores on the distal humerus with surface evaluation (P < 0.05). The carcasses of LRFI pigs were heavier with a higher lean meat percentage (P < 0.001). Most OC scores were unaffected by FA. Only the OC scores of the distal femur (slice method) were higher with increased activity in LRFI pigs, whereas they were lower in HRFI pigs (P < 0.05). Seric biomarkers of cartilage synthesis and degradation were higher for pigs from the LRFI line, but no correlation could be observed between individual OC scores and cartilage biomarker contents.

The impact of housing conditions on porcine mesenchymal stromal/stem cell populations differ between adipose tissue and skeletal muscle

Background. In pigs, the ratio between lean mass and fat mass in the carcass determines production efficiency and is strongly influenced by the number and size of cells in tissues. During growth, the increase in the number of cells results from the recruitment of different populations of multipotent mesenchymal stromal/stem cells (MSCs) residing in the tissues. We hypothesized that the impact of hygiene of housing conditions during growth on the proportions of MSCs in adipose tissue and skeletal muscle may differ between pigs with different residual feed intake (RFI), a measure of feed efficiency. Methods. At the age of 12 weeks, Large White pigs from two lines divergently selected for low and high RFI were housed in two contrasting hygiene conditions (good vs poor). After six weeks, pigs were slaughtered (n = 30; 5-9/group). Samples of subcutaneous adipose tissue and longissimus skeletal muscle were collected, and cells from the stromal vascular fraction (SVF), which includes mesenchymal stromal/stem cell populations, were isolated from each tissue. Adipose and muscle cell populations from the SVF were phenotyped by flow cytometry using antibodies that targeted different cell surface markers (CD45 to separate hematopoietic cells from MSCs; CD34, CD38, CD56 and CD140a to identify MSC populations with adipogenic and/or myogenic potential). Results. Adipose tissue and muscle shared some common MSC populations although MSC diversity was higher in muscle than in adipose tissue. In muscle, the CD45-CD56+CD34-CD140a+ and CD45-CD56+CD34+CD140a+ cell populations were abundant. Of these two cell populations, only the proportions of CD45-CD56+CD34+CD140a+ cells increased (P ≤ 0.05) in pigs housed in poor hygiene as compared with pigs in good hygiene conditions. For the CD45-CD56-CD34- cell population, present in low proportion, there was an interaction between hygiene condition and genetic line (P < 0.05) with a decrease in low RFI pigs housed in poor hygiene conditions. In adipose tissue, the two abundant MSC populations were CD45-CD56-CD34- and CD45-CD56+CD34-. The proportion of CD45-CD56-CD34- cells increased (P < 0.05) whereas the proportion of CD45-CD56+CD34- tended to decrease (P < 0.1) in pigs housed in poor conditions. This study shows that the proportions of some MSC populations were affected by hygiene of housing conditions in a tissue-dependent manner in pigs of both RFI lines. It suggests that MSCs may play a significant role in adipose tissue and skeletal muscle homeostasis and may influence later growth and body composition in growing animals.

Novel Observations of Peroxiredoxin-2 Profile and Protein Oxidation in Skeletal Muscle From Pigs of Differing Residual Feed Intake and Health Status

This study’s objective was to determine the impact of a dual respiratory and enteric bacterial health challenge on the antioxidant protein peroxiredoxin-2 (Prdx-2) profile and protein oxidation in the skeletal muscle of pigs from 2 lines that were divergently selected for residual feed intake (RFI). The hypotheses were that (1) differences exist in the Prdx-2 profile between 2 RFI lines and infection status and (2) muscle from less efficient high-RFI and health-challenged pigs have greater cellular protein oxidation. Barrows (50 ± 7 kg, N = 24) from the 11th generation of the high-RFI (n = 12) and low-RFI (n = 12) Iowa State University lines were used. Pigs (n = 6 per line) were inoculated with Mycoplasma hyopneumoniae and Lawsonia intracellularis (MhLI) on day 0 post infection to induce a respiratory and enteric health challenge. Uninoculated pigs served as controls (n = 6 per line). Necropsy was at 21 d post infection. Sarcoplasmic protein oxidation, various forms of Prdx-2, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) content were determined. Neither RFI line nor infection status significantly affected protein carbonylation. Under nonreducing conditions, MhLI pigs had a greater amount of a slower-migrating GAPDH band (P = 0.017), indicating oxidative modification. Regardless of health status, the low-RFI pigs had less total Prdx-2 (P = 0.035), Prdx-2 decamer (P = 0.0007), and a higher ratio of hyperoxidized peroxiredoxin relative to Prdx-2 (P = 0.028) than the high-RFI pigs. The increased pool of active Prdx-2 in high-RFI pigs suggests greater oxidative stress in muscle in high- versus low-RFI pigs. The increase in oxidized GAPDH seen in muscle from MhLI pigs—particularly the high-RFI MhLI pigs—may be a response to the greater oxidative stress in the high-RFI MhLI. This work suggests that antioxidant proteins are important in growth and health-challenge situations.

Selection for feed efficiency elicits different postprandial plasma metabolite profiles in response to poor hygiene of housing conditions in growing pigs.

This study was conducted to compare postprandial plasma concentrations of insulin, energy-related metabolites, and amino acids measured after a 6-week challenge consisting of exposure to good or poor hygiene of housing conditions of 24 growing pigs divergently selected for low-RFI (LRFI) and high-RFI (HRFI). Blood indicators of immune responses were assessed from samples collected before 0 (W0), and 3 (W3), and 6 weeks (W6) after pigs transfer to their respective hygiene of housing conditions. Plasma haptoglobin concentrations and blood neutrophil granulocyte numbers were greater in poor than in good hygiene of housing conditions at W3. Plasma concentrations of total immunoglobulin G were greater (p = 0.04) in poor than in good hygiene of housing conditions at W6. At W6, pigs were fitted with an intravenous catheter for serial blood samplings. Low-RFI pigs had greater insulin (p < 0.001) and lower triglyceride (p = 0.04) average plasma concentrations than HRFI pigs in both conditions. In poor hygiene of housing conditions, the peaks of insulin and glucose were observed earlier and that of insulin was greater in LRFI than in HRFI pigs. Irrespective of genetic line, average plasma concentrations of histidine, isoleucine, leucine, methionine, threonine, valine, and alanine were greater in poor compared with good hygiene of housing conditions. Only HRFI pigs had greater lysine, asparagine, proline, and tyrosine plasma concentrations in poor than in good hygiene of housing conditions. Conversely, arginine, tryptophan, proline, and tyrosine plasma concentrations were lower only for LRFI pigs housed in poor hygiene conditions. Our results suggest that, contrary to HRFI, LRFI pigs increase or maintain their utilization of tryptophan, arginine, and lysine when housed in poor hygiene conditions. This indicates that this difference may contribute to the better capacity of LRFI to cope with poor hygiene of housing conditions.

Identification of the relationship between the gut microbiome and feed efficiency in a commercial pig cohort.

Feed efficiency (FE) is an economically important trait in pig production. Gut microbiota plays an important role in energy harvest, nutrient metabolism, and fermentation of dietary indigestible components. Whether and which gut microbes affect FE in pigs are largely unknown. Here, a total of 208 healthy Duroc pigs were used as experimental materials. Feces and serum samples were collected at the age of 140 d. We first performed 16S rRNA gene and metagenomic sequencing analysis to investigate the relationship between the gut microbiome and porcine residual feed intake (RFI). 16S rRNA gene sequencing analysis detected 21 operational taxonomic units showing the tendency to correlation with the RFI (P < 0.01). Metagenomic sequencing further identified that the members of Clostridiales, e.g., Ruminococcus flavefaoiens, Lachnospiraceae bacterium 28-4, and Lachnospiraceae phytofermentans, were enriched in pigs with low RFI (high-FE), while 11 bacterial species including 5 Prevotella spp., especially, the Prevotella copri, had higher abundance in pigs with high RFI. Functional capacity analysis suggested that the gut microbiome of low RFI pigs had a high abundance of the pathways related to amino acid metabolism and biosynthesis, but a low abundance of the pathways associated with monosaccharide metabolism and lipopolysaccharide biosynthesis. Serum metabolome and fecal short-chain fatty acids were determined by UPLC-QTOF/MS and gas chromatography, respectively. Propionic acid in feces and the serum metabolites related to amino acid metabolism were negatively correlated with the RFI. The results from this study may provide potential gut microbial biomarkers that could be used for improving FE in pig production industry.

Transcriptome Analysis of Skeletal Muscle in Pigs with Divergent Residual Feed Intake Phenotypes.

Residual feed intake (RFI) is defined as the difference between the observed and expected feed intake for maintenance and growth requirements. In this study, the expression profiles of mRNAs and long noncoding RNAs (lncRNAs) from skeletal muscle in Duroc pigs with divergent RFI phenotypes were investigated by Illumina sequencing. Finally, a total of 2195 annotated lncRNAs and 1976 novel lncRNAs were obtained. About 210 mRNAs and 43 lncRNAs were differentially expressed among high and low RFI pigs. The differentially expressed mRNAs were potentially involved in the biological processes of lipid metabolism, extracellular matrix organization, cell proliferation, and cell adhesion. The lipolysis in skeletal muscle was increased in high RFI pigs, suggesting that high RFI pigs might need more energy than low RFI pigs. However, skeletal muscle development was increased in low RFI pigs. These results suggested that low RFI pigs might be more efficient in energy utilization during skeletal muscle growth. The function of lncRNA was also analyzed by target prediction. Nine lncRNAs might be candidate lncRNAs for the determination of RFI phenotype, by the regulation of the biological processes of lipid metabolism, cell proliferation, and cell adhesion. This study should facilitate a further understanding of the molecular mechanism for the determination of RFI phenotype in pigs.

Colonic microbiome profiles for improved feed efficiency can be identified despite major effects of farm of origin and contemporary group in pigs

While feed efficiency (FE) is a trait of great economic importance to the pig industry, the influence of the intestinal microbiome in determining FE is not well understood. The objective of this experiment was to determine the relative influence of FE and farm of birth on the pig colonic microbiome. Animals divergent in residual feed intake (RFI) were sourced from two geographically distinct locations (farms A + B) in Ireland. The 8 most efficient (low RFI (LRFI)) and 8 least efficient (high RFI, (HRFI)) pigs from farm A and 12 LRFI and 12 HRFI pigs from farm B were sacrificed. Colonic digesta was collected for microbial analysis using 16S ribosomal RNA gene sequencing and also for volatile fatty acid analysis. The α-diversity differed between the farms in this study, with pigs from farm A having greater diversity based on Shannon and InvSimpson measures compared to pigs from farm B (P < 0.05), with no difference identified in either Chao1 or observed measures of diversity (P > 0.05). In the analysis of β-diversity, pigs clustered based on farm of birth rather than RFI. Variation in the management of piglets, weight of the piglets, season of the year, sanitary status and dam dietary influence could potentially be causative factors in this large variation between farms. However, despite significant variation in the microbial profile between farms, consistent taxonomic differences were identified between RFI groups. Within the phylum Bacteroidetes, the LRFI pigs had increased abundance of BS11 (P < 0.05) and a tendency toward increased Bacteroidaceae (P < 0.10) relative to the HRFI group. At genus level, the LRFI pigs had increased abundance of Colinsella (P < 0.05), a tendency toward increased Bacteroides and CF231 (P < 0.10). At species level, Ruminococcus flavefaciens had increased abundance in the LRFI compared to the HRFI animals. In conclusion, while farm of birth has a substantial influence on microbial diversity in the pig colon, a microbial signature indicative of FE status was apparent.

Effect of a dual enteric and respiratory pathogen challenge on swine growth, efficiency, carcass composition, and pork quality1.

The objective of this study was to determine the influence of a dual respiratory and enteric pathogen challenge on growth performance, carcass composition, and pork quality of high and low feed efficient pigs. Pigs divergently selected for low and high residual feed intake (RFI, ~68 kg) from the 11th generation of Iowa State University RFI project were used to represent high and low feed efficiency. To elicit a dual pathogen challenge, half of the pigs (n = 12/line) were inoculated with Mycoplasma hyopneumoniae (Mh) and Lawsonia intracellularis (MhLI) on days post-inoculation (dpi) 0. Pigs in a separate room of the barn were not inoculated and used as controls (n = 12/RFI line). Pigs were weighed and feed intake was recorded to calculate ADG, ADFI, and G:F for the acclimation period (period 1: dpi -21 to 0), during peak infection (period 2: dpi 0 to 42), and during the remaining growth period to reach market weight (period 3: dpi 42 to harvest). At ~125 kg, pigs were harvested using standard commercial procedures. Carcasses were evaluated for composition (weight, fat free lean, loin eye area, 10th rib fat depth) and meat quality (pH decline, temperature decline, Hunter L, a, and b, subjective color and marbling, star probe, drip loss, cook loss, proximate composition, and desmin degradation). Challenged pigs had lesser ADFI than controls during period 2 (P < 0.05), but had greater ADG and G:F during period 3 (P < 0.05). Selection for feed efficiency did not result in a differential response to MhLI (P > 0.05). Loin chops from the less feed efficient, high RFI pigs, had greater drip loss, greater cook loss, lesser moisture content, greater Hunter L values, and greater Hunter b values (P < 0.05) than loin chops from low RFI pigs. Infection status did not significantly affect carcass composition or pork quality traits (P > 0.05). These results indicate that a MhLI challenge early in growth did not significantly affect ultimate carcass composition or meat quality traits. Selection for greater feed efficiency in pigs did not affect their response to pathogenic challenge.

Tissue-specific responses of antioxidant pathways to poor hygiene conditions in growing pigs divergently selected for feed efficiency

BackgroundPoor hygiene of housing induces a systemic inflammatory response. Because inflammation and oxidative stress are processes that can sustain each other, the ways pigs are able to activate their antioxidant defenses are critical for production performance and health during periods when the immune system is solicited. Selection for production performance can also influence reactive oxygen species (ROS) production and expression levels of genes involved in cellular response to oxidative stress in different tissues. To establish the extent by which poor hygiene and selection for feed efficiency affected redox status, pigs divergently selected for residual feed intake (RFI) were housed in poor or good hygiene during 6 weeks. At the end, blood was collected in all pigs, and half of them were killed for tissue sampling. The remaining pigs were reared in good hygiene conditions during a recovery period of 7–8 weeks.ResultsAt week 6, poor hygiene was associated with a lower total antioxidant capacity assessed by plasma ferric reducing ability in all pigs, and with greater plasma levels of hydrogen peroxides in the high RFI pigs (less efficient). Adipose tissue of high RFI pigs exhibited higher activities of catalase and glutathione reductase, and greater thiobarbituric acid reactive substances (TBARS) concentrations when compared with the low RFI pigs (more efficient). Poor hygiene conditions activated the antioxidant enzymes activities (glutathione reductase, superoxide dismutase and catalase) in adipose tissue of both lines, but led to higher ROS production by mature adipocytes isolated from the high RFI pigs only. In liver and muscle, there were only minor changes in antioxidant molecules due to genetics and hygiene conditions. After the resilience period, adipose tissue of pigs previously challenged by poor hygiene maintained higher antioxidant enzyme activities, and for the high RFI line, displayed higher TBARS concentrations.ConclusionsPigs selected for improved feed efficiency showed a lower susceptibility to oxidative stress induced by poor hygiene conditions. This could led to a lower inflammatory response and less impaired growth when these pigs are facing sanitary challenges during the production period.

Machine learning applied to transcriptomic data to identify genes associated with feed efficiency in pigs

BackgroundTo date, the molecular mechanisms that underlie residual feed intake (RFI) in pigs are unknown. Results from different genome-wide association studies and gene expression analyses are not always consistent. The aim of this research was to use machine learning to identify genes associated with feed efficiency (FE) using transcriptomic (RNA-Seq) data from pigs that are phenotypically extreme for RFI.MethodsRFI was computed by considering within-sex regression on mean metabolic body weight, average daily gain, and average backfat gain. RNA-Seq analyses were performed on liver and duodenum tissue from 32 high and 33 low RFI pigs collected at 153 d of age. Machine-learning algorithms were used to predict RFI class based on gene expression levels in liver and duodenum after adjusting for batch effects. Genes were ranked according to their contribution to the classification using the permutation accuracy importance score in an unbiased random forest (RF) algorithm based on conditional inference. Support vector machine, RF, elastic net (ENET) and nearest shrunken centroid algorithms were tested using different subsets of the top rank genes. Nested resampling for hyperparameter tuning was implemented with tenfold cross-validation in the outer and inner loops.ResultsThe best classification was obtained with ENET using the expression of 200 genes in liver [area under the receiver operating characteristic curve (AUROC): 0.85; accuracy: 0.78] and 100 genes in duodenum (AUROC: 0.76; accuracy: 0.69). Canonical pathways and candidate genes that were previously reported as associated with FE in several species were identified. The most remarkable pathways and genes identified were NRF2-mediated oxidative stress response and aldosterone signalling in epithelial cells, the DNAJC6, DNAJC1, MAPK8, PRKD3 genes in duodenum, and melatonin degradation II, PPARα/RXRα activation, and GPCR-mediated nutrient sensing in enteroendocrine cells and SMOX, IL4I1, PRKAR2B, CLOCK and CCK genes in liver.ConclusionsML algorithms and RNA-Seq expression data were found to provide good performance for classifying pigs into high or low RFI groups. Classification was better with gene expression data from liver than from duodenum. Genes associated with FE in liver and duodenum tissue that can be used as predictive biomarkers for this trait were identified.

Responsiveness of swine divergently selected for feed efficiency to exogenous adrenocorticotropic hormone and glucose challenges

Increasing the feed efficiency of lean tissue gains is an important goal for improving sustainable pork production and profitability for swine producers. To study feed efficiency, genetic selection based on residual feed intake (RFI) was used to create two divergent lines. Low-RFI pigs consume less feed for equal weight gain compared with their less-efficient, high-RFI counterparts. As cortisol and insulin are important energy control and growth regulators, our objective was to evaluate the role of the adrenocorticotropic hormone (ACTH)-cortisol and the glucose-insulin axes in pigs divergently selected for RFI. Adrenocorticotropic hormone (0.2 IU/kg BW)–stimulated cortisol and non-esterified fatty acids (NEFA) concentrations and intravenous glucose tolerance test (IVGTT; 0.25 g/kg BW)–stimulated glucose, insulin, and NEFA concentrations were assessed in six low-RFI and six high-RFI gilts (68 ± 5.2 kg). Before the ACTH challenge, low-RFI gilts tended to have less baseline plasma cortisol (P = 0.08) but no difference in NEFA concentrations (P = 0.63) compared with high-RFI gilts. After the ACTH challenge, low-RFI gilts had less cortisol (P = 0.04) and NEFA concentrations (P = 0.05) compared with high-RFI gilts. Glucose, insulin, and NEFA concentrations did not differ between genetic lines before the IVGTT. After glucose infusion, low-RFI gilts had greater insulin concentrations (P = 0.003) but did not differ in glucose or NEFA concentrations compared with high-RFI gilts. These results indicate that genetic selection for reduced RFI (improved feed efficiency) resulted in less stress responsiveness and an increase in insulin after glucose infusion. These data have implications for identifying and selecting more feed efficient pigs and for understanding the physiological mechanisms underlying feed efficiency.

Responses to weaning in two pig lines divergently selected for residual feed intake depending on diet.

Weaning is a stress every piglet has to face. It is a main cause of antibiotic uses due to digestive disorders. In this study, response to weaning was analyzed in pigs from two lines divergently selected for residual feed intake (RFI) during growth. A total of 132 pigs from each line, housed per line and diet in conventional postweaning units of 12 castrated males and 12 females, were fed either a conventional control (two successive diets) or a complex (three successive diets) dietary sequence during the postweaning period (4 to 10 wk of age). BWs were recorded at weaning (days 0 and 28 of age), days 1, 2, 6, 12, 19, 26, and 42 (10 wk of age), and at 23 wk of age. Feces texture was examined before weaning (day -1), at day 1, 2, 6, 12, and 19. Feed intake was recorded at pen level from days 0 to 42 after weaning, and individually thereafter. Plasma was collected after blood samplings at days -1, 6, 19, and 42 on half of the piglets: all piglets of a given sex in each pen were sampled, to achieve a balanced number across factors. Pigs of the low RFI (LRFI) line were heavier at weaning, had greater glucose concentration, and lower levels of diarrhea at days 1 and 2 than pigs from the high RFI (HRFI) line (P < 0.01). At day 42, there was no BW difference between lines, and G:F ratio did not differ between lines (P = 0.40). The LRFI pigs had lower feed intake and growth rate from day 0 to day 19 (P < 0.005), and greater plasma concentration of non-esterified fatty acid (P < 0.001), indicating an increased mobilization of body lipids and proteins immediately after weaning compared with HRFI pigs. They also had greater levels of diarrhea at day 6 (22% for LRFI vs. 14% for HRFI, P = 0.002), but the concentration of plasma haptoglobin did not indicate acute inflammation. The complex diet sequence improved feed intake and growth, and reduced diarrhea, mainly in the LRFI line (P < 0.001). To conclude, pigs from the LRFI line were more negatively affected by weaning stress, but managed to recover afterwards. The complex diet sequence ameliorated some of the negative effects that weaning had on the LRFI pigs, but limited effects of nursery period feeding sequence on growth performance were observed during the growing-finishing period.

Metabolic adaptation of pigs to a Mycoplasma hyopneumoniae and Lawsonia intracellularis dual challenge.

Respiratory and enteric pathogens such as Mycoplasma hyopneumoniae (Mh) and Lawsonia intracellularis (LI) reduce lean accretion and feed efficiency (FE) in growing pigs. However, the metabolic mechanism by which this occurs is still unknown. Therefore, the primary aim of this study was to examine the metabolic adaptation of pigs presented with a dual Mh and LI challenge (MhLI). A secondary objective was to examine if selection for high FE, modeled by selection for low residual feed intake (RFI), alters molecular response to disease. Using a 2 × 2 factorial design, 6 littermate pairs from a high RFI (HRFI) and 6 littermate pairs from a low RFI (LRFI) line (barrows, 66 ± 2 kg BW) were selected, with 1 pig from each pair assigned to individual pens in either the challenge or the nonchallenge (control) rooms (n = 6 barrows per line/challenge). On days post inoculation (dpi) 0, MhLI pigs were inoculated intragastrically with LI and intratracheally with Mh. Pig and feeder weights were recorded at dpi 0, 7, 14, and 21. On dpi 21, pigs were euthanized and tissues and blood were collected. Markers of oxidative stress, skeletal muscle metabolism and proteolysis, and liver gluconeogenesis were evaluated to determine the effects of MhLI, RFI line, and their interaction. The interaction of line and challenge was not significant (P > 0.05) for any measure. Overall, MhLI pigs had lower ADG (38%, P < 0.001), ADFI (25%, P < 0.001), and G:F (19%, P = 0.012) compared with controls. As expected, LRFI pigs had lower ADFI (P = 0.028) for the same ADG, giving them greater G:F (P = 0.021) than HRFI pigs. Challenged pigs had greater reactive oxygen species (ROS) production in the LM and liver (P < 0.10) but did not have greater skeletal muscle proteolysis. Liver gluconeogenesis was also not upregulated (P > 0.05) due to MhLI. These results provide further evidence that selection for LRFI does not negatively affect response to disease. In addition, these results suggest that postabsorptive metabolic functions are altered due to MhLI challenge. The MhLI challenge induced mitochondrial dysfunction, evident by greater ROS production, and caused pigs to favor glycolytic energy generation. However, skeletal muscle proteolysis and liver gluconeogenesis were not upregulated during MhLI challenge. These data suggest that during mild disease stress, pigs can meet energy demands without reliance on nutrient mobilization and gluconeogenesis.

Interactions between metabolically active bacteria and host gene expression at the cecal mucosa in pigs of diverging feed efficiency.

Little is known about the role of the gut mucosal microbiota and microbe-host signaling in the variation of pig's feed efficiency (FE). This study therefore aimed to investigate the FE-related differences in the metabolically active mucosal bacterial microbiota and expression of genes for innate immune response, barrier function, nutrient uptake, and incretins in the cecum of finishing pigs. Pigs (n = 72) were ranked for their residual feed intake (RFI; metric for FE) between days 42 and 91 postweaning and were stratified within litter and sex into high (HRFI; n = 8) and low RFI (LRFI; n = 8). Cecal mucosa and digesta were collected on day 137-141 of life. After isolating total RNA from the mucosa, the RNA was transcribed into cDNA which was used for gene expression analysis, total bacterial quantification, and high-throughput sequencing (Illumina MiSeq) of the hypervariable V3-V4 region of the 16S rRNA gene. The RFI differed by 2.1 kg between low RFI (LRFI; good FE) and high RFI (HRFI; poor FE) pigs (P < 0.001). The cecal mucosa was mainly colonized by Helicobacteraceae, Campylobacteraceae, Veillonellaceae, Lachnospiraceae, and Prevotellaceae. Despite the lack of differences in microbial diversity and absolute abundance, RFI-associated compositional differences were found. The predominant genus Campylobacter tended (P < 0.10) to be 0.4-fold more abundant in LRFI pigs, whereas low abundant Escherichia/Shigella (P < 0.05), Ruminobacter (P < 0.05), and Veillonella (P < 0.10) were 3.4-, 6.6-, and 4.4-fold less abundant at the cecal mucosa of LRFI compared to HRFI pigs. Moreover, mucin 2 and zona occludens-1 were less expressed (P < 0.05) in the cecal mucosa of LRFI compared to HRFI pigs. Cecal mucosal expression of monocarboxylate transporter-1, glucagon-like peptide-1, and peptide YY further tended (P < 0.10) to be downregulated in LRFI compared to HRFI pigs, indicating an enhanced VFA uptake and signaling in HRFI pigs. Sparse partial least square regression and relevance networking support the hypothesis that certain mucosal bacteria and luminal microbial metabolites were more associated than others with differences in RFI and cecal gene expression. However, present results do not allow the determination of whether mucosal bacterial changes contributed to variation in FE or were rather a consequence of FE-related changes in the pig's physiology or feeding behavior.

Effect of lower-energy, higher-fiber diets on pigs divergently selected for residual feed intake when fed higher-energy, lower-fiber diets.

Residual feed intake (RFI) is the difference between observed and predicted feed intake of an animal, based on growth and maintenance requirements. In Yorkshire pigs, divergent selection for increased (Low RFI) and decreased (High RFI) RFI was carried out over 10 generations (G) while feeding a corn- and soybean-meal-based, higher-energy, lower-fiber (HELF) diet. In G8 to G10, representing 4 replicates, barrows and gilts (n = 649) of the RFI lines were fed the HELF diet and a diet incorporating coproducts that were lower in energy and higher in dietary fiber (LEHF). The diets differed in ME, 3.32 vs. 2.87 Mcal/kg, and in neutral detergent fiber (NDF), 9.4% vs. 25.9%, respectively. The impact of the LEHF diet on 1) performance and growth, 2) diet digestibility, 3) genetic parameter estimates, and 4) responses to selection for RFI, when fed the HELF, was assessed. In general, the LEHF diet reduced the performance of both lines. When fed the HELF diet, the Low RFI pigs had lower (P < 0.05) ADFI (-12%), energy intake (-12%), ADG (-6%), and backfat depth (-12%); similar (P > 0.05) loin muscle area (LMA; +5%); and greater (P < 0.05) feed efficiency (i.e., 8% higher G:F and 7% lower RFI) than the High RFI line. These patterns of line differences were still present under the LEHF diet but differences for ADFI (-11%), energy intake (-10%), G:F (+2%), and RFI (-6%) were reduced compared to the HELF diet. Apparent total tract digestibility (ATTD) of the HELF and LEHF diets was assessed using 116 barrows and gilts from G8. When fed the HELF diet, ATTD of DM, GE, N, and NDF were similar between lines (P ≥ 0.27), but when fed the LEHF diet, the Low RFI pigs had greater digestibility (7%, 7%, 10%, and 32%) than the High RFI line (P ≤ 0.04). To measure responses to selection for RFI and estimate genetic parameters, data from all 10 generations were used (HELF; n = 2,310; LEHF, n = 317). Heritability estimates of performance traits ranged from 0.19 to 0.63, and genetic correlations of traits between diets were high and positive, ranging from 0.87 (RFI) to 0.99 (LMA). By G10, RFI in the Low RFI line was 3.86 and 1.50 genetic SD lower than in the High RFI line when fed the HELF and LEHF diets, respectively. Taken together, the results of this study demonstrate that responses to selection for RFI when fed a HELF diet are not fully realized when pigs are fed an extremely LEHF diet. Thus, feeding diets that differ from those used for selection may not maximize genetic potential for feed efficiency.

Impact of hygiene of housing conditions on performance and health of two pig genetic lines divergent for residual feed intake

Pigs selected for high performance may be more at risk of developing diseases. This study aimed to assess the health and performance of two pig lines divergently selected for residual feed intake (RFI) (low RFI (LRFI) v. high RFI (HRFI)) and housed in two contrasted hygiene conditions (poor v. good) using a 2×2 factorial design (n=40/group). The challenge period (Period 1), started on week zero (W0) when 12-week-old pigs were transferred to good or poor housing conditions. At week 6 (W6), half of the pigs in each group were slaughtered. During a recovery period (Period 2) from W6 to W13 to W14, the remaining pigs (n=20/group) were transferred in good hygiene conditions before being slaughtered. Blood was collected every three (Period 1) or 2 weeks (Period 2) to assess blood indicators of immune and inflammatory responses. Pulmonary lesions at slaughter and performance traits were evaluated. At W6, pneumonia prevalence was greater for pigs housed in poor than in good conditions (51% v. 8%, respectively, P<0.001). Irrespective of hygiene conditions, lung lesion scores were lower for LRFI pigs than for HRFI pigs (P=0.03). At W3, LRFI in poor conditions had the highest number of blood granulocytes (hygiene×line, P=0.03) and at W6, HRFI pigs in poor conditions had the greatest plasma haptoglobin concentrations (hygiene×line, P=0.02). During Period 1, growth rate and growth-to-feed ratio were less affected by poor hygiene in LRFI pigs than in HRFI pigs (hygiene×line, P=0.001 and P=0.02, respectively). Low residual feed intake pigs in poor conditions ate more than the other groups (hygiene×line, P=0.002). Irrespective of the line, fasting plasma glucose concentrations were higher in poor conditions, whereas fasting free fatty acids concentrations were lower than in good conditions. At the end of Period 2, pneumonia prevalence was similar for both housing conditions (39% v. 38%, respectively). During Period 2, plasma protein concentrations were greater for pigs previously housed in poor than in good conditions during Period 1. Immune traits, gain-to-feed ratio, BW gain and feed consumption did not differ during Period 2. Nevertheless, at W12, BW of HRFI previously housed in poor conditions was 13.4 kg lower than BW of HRFI pigs (P<0.001) previously housed in good conditions. In conclusion, health of the most feed efficient LRFI pigs was less impaired by poor hygiene conditions. This line was able to preserve its health, growth performance and its feed ingestion to a greater extent than the less efficient HRFI line.

Novel Observations of Peroxiredoxin-2 Profile and Protein Oxidation in Skeletal Muscle from Pigs that Differ In Residual Feed Intake and Health Status

ObjectivesEfficiency of pork production is critical as demand for animal protein rises with increases in global populations and income. Feed efficiency and response to disease challenge are affected by stressors that increase reactive oxygen species in muscle. Reactive oxygen species are detrimental to growth, as they require the expenditure of energy for tissue repair that could be used for weight gain. Peroxiredoxin-2 (Prdx-2) is an antioxidant enzyme that converts hydrogen peroxide to water using reactive cysteines, thus mitigating oxidation. Prdx-2 can exist in multiple oxidation states (reduced, oxidized, and hyperoxidized) and quantenary structures (dimer and decamer). Prdx-2 concentration is greater in more color stable muscles and may be an indicator of tenderness. The objective of this study was to determine differences in Prdx-2 profile and protein oxidation between pigs that differed in feed efficiency and health status. Materials and MethodsPigs selected for differing feed efficiency based on residual feed intake (RFI) were used in a 2 × 2 factorial design for this study. At 50 ± 7 kg in body weight, high RFI (less efficient) and low RFI (more efficient) barrows were distributed between 2 rooms in the same building. One room was inoculated with a dual respiratory/enteric bacterial health challenge. At 21 d post-infection (projected peak of illness), pigs were necropsied (total = 24, n = 6 per group) and longissimus muscle samples were collected. Reduced and nonreduced sarcoplasmic protein samples (β-mercaptoethanol) were used for western blot analyses. Reduced samples were used to quantify total Prdx-2, while nonreduced samples were used to evaluate hyperoxidized peroxiredoxin, Prdx-2 decamer, and Prdx-2 profile in nonreducing gels (2 distinct bands analyzed). Carbonyl content and diagonal gel electrophoresis were performed on sarcoplasmic protein samples to determine protein oxidation. Statistical analysis was performed using SAS v. 9.4 (SAS Inst. Inc., Cary, NC) with fixed effects of RFI line, infection status, and RFI line × infection status interaction. Gel repetition and necropsy day were included as random effects. ResultsInfection status was significant for the Prdx-2 band comparison in nonreducing gels (P = 0.014) indicating potential differences in posttranslational modifications. Compared to low RFI pigs, high RFI pigs had greater total Prdx-2 (P = 0.035), greater Prdx-2 decamer (P = 0.0007), greater Prdx-2 in the second band of nonreducing gels (P = 0.0006), and less hyperoxidized peroxiredoxin relative to the total immunoreactive protein (P = 0.028). An interaction existed between RFI line and infection status for the Prdx-2 band comparison (P = 0.02), with high RFI challenged pigs having greater ratios, and high RFI control pigs having lesser ratios relative to low RFI pigs. Visual analysis of diagonal gels showed more proteins with intermolecular disulfide formation in low RFI muscle samples. No significant difference was seen for carbonyl content. ConclusionDifferences exist in Prdx-2 profile based on RFI line and infection status. High and low RFI pigs may respond differently to health challenges at the molecular level based on differences in antioxidant protein response. By defining Prdx-2 in livestock muscle, we may be better able to manage animals to meet demand for meat without increasing inputs.

Effects of Diet and Genetics on Growth Performance of Pigs in Response to Repeated Exposure to Heat Stress.

Heat stress (HS) is one of the costliest issues in the U.S. pork industry. Aims of the present study were to determine the consequences of repeated exposure to HS on growth performance, and the effects of a high fiber diet, the genetic potential for high lean tissue accretion, and the genetic potential for residual feed intake (RFI) on resilience to HS. Barrows (n = 97) from three genetic lines (commercial, high RFI, low RFI) where subjected three times to a 4-day HS treatment (HS1, HS2, and HS3) which was preceded by a 9-day neutral (TN) adaptation period (TN1) and alternated by 7-day periods of neutral temperatures (TN2, TN3, and TN4). Body weight gain (BWG), feed intake (FI), feed conversion efficiency (FCE), RFI, and the drop in BWG and FI between TN and HS were estimated for each period, and slaughter traits were measured at the end of TN4. Commercial pigs had lower FI when fed a high fiber diet compared to a regular diet (2.70 ± 0.08 vs. 2.96 ± 0.08 kg/d; P < 0.05), while no differences were found for BWG, RFI or FCE. HS reduced FI, BWG, and FCE, increased RFI, and resulted in leaner pigs that generate smaller carcasses at slaughter. In TN, commercial pigs grew faster than the low and high RFI pigs (1.22 ± 0.06 vs. 0.720 ± 0.05 and 0.657 ± 0.07; P < 0.001) but growth rates were not significantly different between the lines during HS. Growth rates for the low RFI and high RFI pigs were similar both during TN and during HS. Pigs of interest for genetic improvement are those that are able to maintain growth rates during HS. Our results show that response in growth to HS was repeatable over subsequent 4-d HS cycles, which suggests the potential for including this response in the breeding index. The best performing animals during HS are likely those that are not highly superior for growth in TN.