Experimental facility had a greater effect on growth performance, gut microbiome, and metabolome in weaned pigs than feeding diets containing subtherapeutic levels of antibiotics: A case study.

The objective of this study was to define changes in the intestinal metabolome and microbiome associated with growth performance of weaned pigs fed subtherapeutic concentrations of antibiotics. Three experiments with the same antibiotic treatments were conducted on the same research farm but in two different facilities (nursery and wean-finish) using pigs weaned at 20-days of age from the same source herd and genotype, and fed the same diets formulated without antibiotics (NC) or with 0.01% chlortetracycline and 0.01% sulfamethazine (AB). Pigs were weighed and feed disappearance was determined on days (d) 10, 21, and 42 post-weaning to calculate average daily gain (ADG), average daily feed intake (ADFI), and gain:feed (G:F). On d 42, one pig/pen was selected for blood and ileal and cecal content collection. Targeted and untargeted metabolomic profiles were determined in serum and cecal contents using liquid chromatography-mass spectrometry, and composition of bacterial communities in intestinal content samples was determined by sequencing the V4 region of the 16s rRNA gene. Metabolomics and microbiome data were analyzed using diverse multivariate and machine learning methods. Pigs fed AB had significantly greater (P < 0.05) overall ADG and ADFI compared with those fed NC, and pig body weight, ADG, and G:F were also significantly different (P < 0.05) between experiments. Differences (P < 0.05) in serum metabolome along with ileal and cecal microbiome beta diversity were observed between experiments, but there were no differences in microbiome alpha diversity between experiments or treatments. Bacteria from the families Clostridiaceae, Streptomycetaceae, Peptostreptomycetaceae, and Leuconostocaceae were significant biomarkers for the AB treatment. In addition, pigs fed AB had increased serum arginine, histidine, lysine, and phenylalanine concentrations compared with NC. Percentage error from a random forest analysis indicated that most of the variation (8% error) in the microbiome was explained by the facility where the experiments were conducted. These results indicate that facility had a greater effect on growth performance, metabolome, and microbiome responses than feeding diets containing subtherapeutic levels of antibiotics.

Growth Performance, Metabolomics, and Microbiome Responses of Weaned Pigs Fed Diets Containing Growth-Promoting Antibiotics and Various Feed Additives.

The objective of this study was to determine the potential biological mechanisms of improved growth performance associated with potential changes in the metabolic profiles and intestinal microbiome composition of weaned pigs fed various feed additives. Three separate 42 day experiments were conducted to evaluate the following dietary treatments: chlortetracycline and sulfamethazine (PC), herbal blends, turmeric, garlic, bitter orange extract, sweet orange extract, volatile and semi-volatile milk-derived substances, yeast nucleotide, and cell wall products, compared with feeding a non-supplemented diet (NC). In all three experiments, only pigs fed PC had improved (p < 0.05) ADG and ADFI compared with pigs fed NC. No differences in metabolome and microbiome responses were observed between feed additive treatments and NC. None of the feed additives affected alpha or beta microbiome diversity in the ileum and cecum, but the abundance of specific bacterial taxa was affected by some dietary treatments. Except for feeding antibiotics, none of the other feed additives were effective in improving growth performance or significantly altering the metabolomic profiles, but some additives (e.g., herbal blends and garlic) increased (p < 0.05) the relative abundance of potentially protective bacterial genera that may be beneficial during disease challenge in weaned pigs.

Effects of medium chain fatty acids as a mitigation or prevention strategy against porcine epidemic diarrhea virus in swine feed.

Feed has been shown to be a vector for viral transmission. Four experiments were conducted to: 1) determine if medium chain fatty acids (MCFA) are effective mitigants when applied to feed both pre- and post-porcine epidemic diarrhea virus (PEDV) inoculation measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR), 2) evaluate varying levels and combinations of MCFA measured by qRT-PCR, and 3) evaluate selected treatments in bioassay to determine infectivity. In exp. 1, treatments were arranged in a 2 × 2 + 1 factorial with main effects of treatment (0.3% commercial formaldehyde [CF] product, Sal CURB [Kemin Industries, Inc.; Des Moines, IA], or 1% MCFA blend (Blend) of 1:1:1 C6:C8:C10 [PMI, Arden Hills, MN]) and timing of application (pre- or post-inoculation with PEDV) plus a positive control (PC; feed inoculated with PEDV and no treatment). All combinations of treatment and timing decreased detectable PEDV compared with the PC (P < 0.05). Pre-inoculation treatment elicited decreased magnitude of PEDV detection (cycle threshold value) compared with post-inoculation (P = 0.009). Magnitude of PEDV detection was decreased for CF compared with Blend (P < 0.0001). In exp. 2, pre-inoculation treatments consisted of: 1) PC, 2) 0.3% CF, 3 to 5) 0.125% to 0.33% C6:0, 6 to 8) 0.125% to 0.33% C8:0, 9 to 11) 0.125% to 0.33% C10:0, and 12 to 15) 0.125% to 0.66% C5:0. Treating feed with 0.33% C8:0 resulted in decreased (P < 0.05) PEDV detection compared with all other treatments. Increasing concentration of each individual MCFA decreased PEDV detectability (P < 0.042). In exp. 3, pre-inoculation treatments consisted of: 1) PC, 2) 0.3% CF, 3 to 7) 0.25% to 1% Blend, 8 to 10) 0.125% to 0.33% C6:0 + C8:0, 11 to 13) 0.125% to 0.33% C6:0 + C10:0, and 14 to 16) 0.125% to 0.33% C8:0 + C10:0. Treating feed with CF, 0.5% Blend, 0.75% Blend, 1% Blend, all levels of C6:0+C8:0, 0.25% C6:0 + 0.25% C10:0, 0.33% C6:0 + 0.33% C10:0, 0.25% C8:0 + 0.25% C10:0, or 0.33% C8:0 + 0.33% C10:0 elicited decreased detection of PEDV compared with PC (P < 0.05). Increasing concentration of each MCFA combination decreased PEDV detectability (linear, P < 0.012). In exp. 4, feed was treated pre-inoculation with: 1) no treatment (PC), 2) 0.3% CF, 3) 0.5% Blend, or 4) 0.3% C8:0 and analyzed via qRT-PCR and bioassay. Adding 0.5% Blend or 0.3% C8:0 resulted in decreased PEDV compared with PC and only PC resulted in a positive bioassay. Therefore, MCFA can decrease detection of PEDV in feed. Further, inclusion of lower levels of MCFA than previously evaluated are effective against PEDV.

Microbiome profiling of commercial pigs from farrow to finish.

Balanced bacterial communities within the gastrointestinal (GI) tract of animals are a key component of gut health, resulting in optimal performance and the prevention of disease. The purpose of this study was to characterize the commercial pig's baseline bacterial microbiome over time and across anatomical site. Several anatomical sites (duodenum/jejunum, ileum, cecum, and colon) were examined across multiple ages (days 0, 10, 21, 33, 62, 84, and market) for bacterial microbiome structure using 16S rRNA V4 region sequencing with Illumina MiSeq. General trends in the succession of the bacterial microbiome were observed over age, such as increasing populations of Clostridia and decreasing populations of Gammaproteobacteria (P < 0.05). However, apparent disruptions in the microbiome were also observed that did not follow these trends, specifically at sampling 24 h post-weaning where Lactobacillaceae were drastically reduced in relative abundance (P < 0.05). The introduction of solid feed between days 21 and 33 had the greatest overall impact on bacterial community structure as compared with the effects of age, changes in solid feed type, and pig movement. A core bacterial microbiome was identified across all anatomical sites consisting of the dominant operational taxonomic units (OTUs); samples were only differentiated based upon anatomical site when considering less abundant OTUs and differences in relative abundance. When considering mucosal vs. digesta samples from the cecum and ileum, several taxa were of significantly higher relative abundance in the mucosa (P < 0.05), including Anaerovibrio, Bacteroides, Desulfovibrio, Helicobacter, Oscillospira, Phascolarctobacterium, and Prevotella. Correlations between several genus-level taxa and pig weight were observed. Overall, this study provides an expanded view of the dynamic pig GI microbiome from farrow to finish.

Endotoxin decreases 51CrCl3 uptake in early weaned pigs.

Endotoxin, a product of bacterial infections, was used to investigate the effects acute infections on chromium (Cr) uptake in early weaned (14 d of age) pigs. At d 18, pigs were anesthetized, and a 2-mm silastic jugular catheter was inserted and passed subcutaneously to a dorsal position behind the ear. At d 21, pigs were deprived of food for 7 h and injected intraperitoneally with saline or 25 microg endotoxin (lipopolysaccharide from Escherichia coli serotype 0111:B4) per kg body wt suspended in 9 g/L saline. One hour after dosing, an oral dose of 0.7 mCi of 51CrCl3 was given by micropipet. Blood was sampled from the catheter at intervals until necropsy at 8 h after the 51CrCl3 dose. Blood and tissue samples were counted in a gamma counter. 51Cr in blood was significantly lower at 3, 4, 5, 6, and at 8 h after dosing in endotoxin-injected pigs compared to controls. Eight hours after 51CrCl3 dosing, 51Cr retention was significantly lower in the liver, heart, and kidney in endotoxin-treated pigs and tended to be lower in spleen (p<0.06) and in urine (p<0.16) with endotoxin treatment. These data suggest that during acute infection, there might be decreased Cr uptake and retention.