The Role of Salmonella Genomic Island 4 in Metal Tolerance of Salmonella enterica Serovar I 4,[5],12:i:- Pork Outbreak Isolate USDA15WA-1.

Multidrug-resistant (MDR; resistance to >3 antimicrobial classes) Salmonella enterica serovar I 4,[5],12:i:- strains were linked to a 2015 foodborne outbreak from pork. Strain USDA15WA-1, associated with the outbreak, harbors an MDR module and the metal tolerance element Salmonella Genomic Island 4 (SGI-4). Characterization of SGI-4 revealed that conjugational transfer of SGI-4 resulted in the mobile genetic element (MGE) replicating as a plasmid or integrating into the chromosome. Tolerance to copper, arsenic, and antimony compounds was increased in Salmonella strains containing SGI-4 compared to strains lacking the MGE. Following Salmonella exposure to copper, RNA-seq transcriptional analysis demonstrated significant differential expression of diverse genes and pathways, including induction of at least 38 metal tolerance genes (copper, arsenic, silver, and mercury). Evaluation of swine administered elevated concentrations of zinc oxide (2000 mg/kg) and copper sulfate (200 mg/kg) as an antimicrobial feed additive (Zn+Cu) in their diet for four weeks prior to and three weeks post-inoculation with serovar I 4,[5],12:i:- indicated that Salmonella shedding levels declined at a slower rate in pigs receiving in-feed Zn+Cu compared to control pigs (no Zn+Cu). The presence of metal tolerance genes in MDR Salmonella serovar I 4,[5],12:i:- may provide benefits for environmental survival or swine colonization in metal-containing settings.

Identification of C9-C11 unsaturated aldehydes as prediction markers of growth and feed intake for non-ruminant animals fed oxidized soybean oil.

BACKGROUND: The benefits of using the oxidized oils from rendering and recycling as an economic source of lipids and energy in animal feed always coexist with the concerns that diverse degradation products in these oxidized oils can negatively affect animal health and performance. Therefore, the quality markers that predict growth performance could be useful when feeding oxidized oils to non-ruminants. However, the correlations between growth performance and chemical profiles of oxidized oils have not been well examined. In this study, six thermally oxidized soybean oils (OSOs) with a wide range of quality measures were prepared under different processing temperatures and processing durations, including 45 °C-336 h; 67.5 °C-168 h; 90 °C-84 h; 135 °C-42 h; 180 °C-21 h; and 225 °C-10.5 h. Broilers and nursery pigs were randomly assigned to diets containing either unheated control soybean oil or one of six OSOs. Animal performance was determined by measuring body weight gain, feed intake, and gain to feed ratio. The chemical profiles of OSOs were first evaluated by common indicative tests, including peroxide value, thiobarbituric acid reactive substances, p-anisidine value, free fatty acids, oxidized fatty acids, unsaponifiable matter, insoluble impurities, and moisture, and then analyzed by the liquid chromatography-mass spectrometry-based chemometric analysis. RESULTS: Among common quality indicators, p-anisidine value (AnV), which reflects the level of carbonyl compounds, had the greatest inverse correlation with the growth performance of both broilers and pigs, followed by free fatty acids and oxidized fatty acids. Among the 17 aldehydes identified in OSOs, C9-C11 alkenals, especially 2-decenal and 2-undecenal, had stronger inverse correlations (r < - 0.8) with animal performance compared to C5-C8 saturated alkanals, suggesting that chain length and unsaturation level affect the toxicity of aldehydes. CONCLUSIONS: As the major lipid oxidation products contributing to the AnV, individual C9-C11 unsaturated aldehydes in heavily-oxidized oils could function as effective prediction markers of growth and feed intake in feeding non-ruminants.

Impact of viral disease hypophagia on pig jejunal function and integrity.

Pathogen challenges are often accompanied by reductions in feed intake, making it difficult to differentiate impacts of reduced feed intake from impacts of pathogen on various response parameters. Therefore, the objective of this study was to determine the impact of Porcine Reproductive and Respiratory Syndrome virus (PRRSV) and feed intake on parameters of jejunal function and integrity in growing pigs. Twenty-four pigs (11.34 ± 1.54 kg BW) were randomly selected and allotted to 1 of 3 treatments (n = 8 pigs/treatment): 1) PRRSV naïve, ad libitum fed (Ad), 2) PRRSV-inoculated, ad libitum fed (PRRS+), and 3) PRRSV naïve, pair-fed to the PRRS+ pigs' daily feed intake (PF). At 17 days post inoculation, all pigs were euthanized and the jejunum was collected for analysis. At days post inoculation 17, PRRS+ and PF pigs had decreased (P < 0.05) transepithelial resistance compared with Ad pigs; whereas fluorescein isothiocyanate-dextran 4 kDa permeability was not different among treatments. Active glucose transport was increased (P < 0.05) in PRRS+ and PF pigs compared with Ad pigs. Brush border carbohydrase activity was reduced in PRRS+ pigs compared with PF pigs for lactase (55%; P = 0.015), sucrase (37%; P = 0.002), and maltase (30%; P = 0.015). For all three carbohydrases, Ad pigs had activities intermediate that of PRRS+ and PF pigs. The mRNA abundance of the tight junction proteins claudin 2, claudin 3, claudin 4, occludin, and zonula occludens-1 were reduced in PRRS+ pigs compared with Ad pigs; however, neither the total protein abundance nor the cellular compartmentalization of these tight junction proteins differed among treatments. Taken together, this study demonstrates that the changes that occur to intestinal epithelium structure, function, and integrity during a systemic PRRSV challenge can be partially explained by reductions in feed intake. Further, long term adaptation to PRRSV challenge and caloric restriction does reduce intestinal transepithelial resistance but does not appear to reduce the integrity of tight junction protein complexes.

Lack of interactive effects between diet composition and acid addition with drying method on amino acid digestibility values in porcine ileal digesta.

Two experiments were conducted to determine the effect of oven drying (OD) or freeze drying (FD) on apparent ileal digestibility (AID) of AA in diets fed to pigs. In experiment 1, 15 barrows (88.4 ± 6.4 kg) were allotted to either a corn starch-soybean meal (CST), potato starch-soybean meal (PST), or corn-soybean meal (CSBM) diet, over 2 collection periods. Following collection, samples were pooled within pig and subdivided into either OD or FD, resulting in 10 observations per diet by drying-method combination. In experiment 2, 11 barrows (63.3 ± 3.8 kg) were fed a CST diet, and, following collection, samples were pooled within pig and subdivided and either adjusted to pH 4 or remain unadjusted. Subsets of these samples were then subdivided to be either FD or OD, resulting in 11 observations per pH level by drying-method combination. OD was accomplished by drying samples in a forced air oven at either 100 °C (experiment 1) or 75 °C (experiment 2). In experiment 1, there was no diet type by drying-method interactions noted for any of the AA (P > 0.10).OD resulted in a higher AID of AA compared with samples which were FD (P ≤ 0.10), for all AA except for Gly and Tyr. Averaged across all AA, AID of AA was 3.3% greater if the sample was OD compared with FD. Differences in AID of AA among the 3 diets were noted for all AA (P ≤ 0.07), except for Cys (P = 0.33), due to the fact that CST and PST diets only contained soybean meal (SBM) as an AA-providing ingredient while the CSBM diet contained both corn and SBM. Pigs fed the PST diet had greater SID for all AA compared with pigs fed the CST diet (P ≤ 0.05), except for His, Lys, Cys, and Glu. In experiment 2, there were no pH-adjustment by drying-method interactions noted on AID for any of the AA (P > 0.10). Adjusting ileal digesta to a pH of 4.0 had little effect on AID for most of the AA, except for a lowered AID of Arg, His, Lys, Trp, and Ser (P ≤ 0.10). OD resulted in a higher AID for all AA (P ≤ 0.09) except for Ile, Thr, Val, Ala, Asp, Glu, and Gly. Averaged across all AA, the increase in AID of AA was 1.7% greater if the sample was OD compared with FD. On average, OD of ileal digesta resulted in a 2.5% greater estimate of AID of AA compared with samples that were FD, and was not diet-, pH-, or AA-dependent. Because the majority of the data on AA digestibility are based on FD, a bias factor may be necessary to adjust AA digestibility data obtained on an OD-basis relative to an FD-basis for use in feed formulation.

Amino acid composition and digestible amino acid content in animal protein by-product meals fed to growing pigs1.

An industry survey and animal experiment were conducted to evaluate the amino acid (AA) compositional variability and standardized ileal digestibility (SID) of AA in animal protein by-products fed to growing pigs. Animal protein by-product meals (212) were categorized into 8 groupings (blood meal, chicken by-product meal, chicken meal, feather meal, meat and bone meal, meat meal, poultry by-product meal, and poultry meal) and analyzed for total AA. Amino acid analysis among (e.g., Lys in blood meal averaged 9.20% compared with 2.31% for feather meal, DM basis) and within (e.g., Lys range of 1.54% in blood meal and 1.44% in feather meal, DM basis) the by-product classifications varied as expected, but on average the total AA values were similar to that reported in the literature. For the determination of the SID of AA, 15 barrows (average initial and final BW of 31.6 and 78.7 kg, respectively) were fitted with a T-cannula in the distal ileum and allotted to 15 diets over nine 7-d periods, resulting in 9 replications per diet. Pigs were fed a basal diet based on soybean meal and dehulled-degermed corn, 13 diets containing 17.5% animal protein by-product meal to partially replace a portion of the soybean meal and dehulled-degermed corn in the basal diet, or a N-free diet. Pigs were re-allotted to diets based on minimizing the number to times that the N-free diet would precede or follow feeding either of the blood or feather meal diets because of concern with inadequate diet consumption, as well as to prevent diets from being re-fed to the same pig during the next or subsequent periods. Values for the apparent ileal AA digestibility of each diet were determined, adjusted to SID based upon the endogenous AA losses determined by feeding the N-free diet, and the SID of AA in each animal protein by-product meal calculated using the difference procedure. The SID of AA varied among (e.g., SID of Lys averaged 91% in chicken meal but 47% in feather meal) and within (e.g., SID of Lys in three meat and bone meals was 80%, 71%, and 54%) the animal protein by-product meals, as would be expected and are consistent with variation reported in the literature. Overall, the data provide total AA composition for 212 samples and SID of AA for 13 samples of animal protein by-product meals, including data on their variability, which is critical for their use in feed formulation programs.

Impact of porcine reproductive and respiratory syndrome virus on muscle metabolism of growing pigs1.

Porcine reproductive and respiratory syndrome (PRRS) virus is one of the most economically significant pig pathogens worldwide. However, the metabolic explanation for reductions in tissue accretion observed in growing pigs remains poorly defined. Additionally, PRRS virus challenge is often accompanied by reduced feed intake, making it difficult to discern which effects are virus vs. feed intake driven. To account for this, a pair-fed model was employed to examine the effects of PRRS challenge and nutrient restriction on skeletal muscle and liver metabolism. Forty-eight pigs were randomly selected (13.1 ± 1.97 kg BW) and allotted to 1 of 3 treatments (n = 16 pigs/treatment): 1) PRRS naïve, ad libitum fed (Ad), 2) PRRS-inoculated, ad libitum fed (PRRS+), and 3) PRRS naïve, pair-fed to the PRRS-inoculated pigs' daily feed intake (PF). At days postinoculation (dpi) 10 and 17, 8 pigs per treatment were euthanized and tissues collected. Tissues were assayed for markers of proteolysis (LM only), protein synthesis (LM only), oxidative stress (LM only), gluconeogenesis (liver), and glycogen concentrations (LM and liver). Growth performance, feed intake, and feed efficiency were all reduced in both PRRS+ and PF pigs compared with Ad pigs (P < 0.001). Furthermore, growth performance and feed efficiency were additionally reduced in PRRS+ pigs compared with PF pigs (P < 0.05). Activity of most markers of LM proteolysis (µ-calpain, 20S proteasome, and caspase 3/7) was not increased (P > 0.10) in PRRS+ pigs compared with Ad pigs, although activity of m-calpain was increased in PRRS+ pigs compared with Ad pigs (P = 0.025) at dpi 17. Muscle reactive oxygen species production was not increased (P > 0.10) in PRRS+ pigs compared with Ad pigs. However, phosphorylation of protein synthesis markers was decreased in PRRS+ pigs compared with both Ad (P < 0.05) and PF (P < 0.05) pigs. Liver gluconeogenesis was not increased as a result of PRRS; however, liver glycogen was decreased (P < 0.01) in PRRS+ pigs compared with Ad and PF pigs at both time points. Taken together, this work demonstrates the differential impact a viral challenge and nutrient restriction have on metabolism of growing pigs. Although markers of skeletal muscle proteolysis showed limited evidence of increase, markers of skeletal muscle synthesis were reduced during PRRS viral challenge. Furthermore, liver glycogenolysis seems to provide PRRS+ pigs with glucose needed to fuel the immune response during viral challenge.

Mycoplasma hyopneumoniae-Lawsonia intracellularis dual challenge modulates intestinal integrity and function1.

Lawsonia intracellularis (LI) and Mycoplasma hyopneumoniae (Mh) are 2 globally distributed pathogens that cause significant morbidity and mortality in grow-finish pigs. However, mechanisms that reduce growth and feed efficiency during LI and Mh infection are poorly defined. We hypothesized that reductions in performance are partially due to declines in intestinal function and integrity; thus, this study aimed to evaluate intestinal function and integrity of pigs during a 21-d Mh and LI dual challenge (MhLI). Littermate pairs of barrows (48.1 ± 6.7 kg BW) were selected; 1 pig from each pair was assigned to either MhLI challenge or nonchallenge treatments (n = 12). Pigs were individually housed, fed a corn-soybean diet, and allowed to acclimate for 21 d prior to inoculation. On days postinoculation (dpi) 0, MhLI pigs were dual inoculated with LI and Mh. On dpi 21, all pigs were euthanized for ileal and colon tissue collection. Formalin-fixed tissues were clinically scored and morphology analyzed, frozen tissues assayed for digestive enzyme activities, and fresh tissues mounted into modified Ussing Chambers to assess active nutrient transport, barrier integrity, and bacterial translocation. Data were analyzed using the Mixed Procedure of SAS with treatment as a fixed effect, age and start BW as covariates, and litter as a random effect. Compared with controls, MhLI pigs had decreased ADG (38%, P < 0.001), ADFI (25%, P < 0.001), and G:F (19%, P = 0.012). The MhLI dual challenge did not alter ileum morphology or transepithelial resistance (P > 0.10); however, ex vivo mucosal to serosal translocation of S. Typhimurium in the colon was increased (60%, P = 0.003) in MhLI pigs compared with controls. Additionally, MhLI pigs had increased ileal glucose transport (30%, P = 0.05) and decreased sucrase activity (30%, P = 0.049) compared with controls. This MhLI challenge antagonized intestinal function and integrity, and this may be a contributing factor to reduced pig performance.

Evaluating nursery pig responses to in-feed sub-therapeutic antibiotics.

Antibiotics have been used for over 60 years by the swine industry to improve growth performance and feed efficiency. With rising concerns over antimicrobial resistance and government restrictions such as the Veterinary Feed Directive on usage of in-feed antibiotics, alternatives to feeding antibiotic growth promoters (AGPs) to nursery pigs are needed. However, the mechanism of action by which AGPs work is poorly understood. Thus, the objective of this study was to investigate the mechanisms of action by which AGPs increase nursery pig performance. Over two replicates, 24 weaned pigs (6.75 ± 0.75 kg body weight) were randomly allotted to either control (CON, n = 12) or sub-therapeutic antibiotic (sCTC, n = 12) treatments and housed individually. A 2-phase corn-soybean-based nursery diet was fed, with the sCTC diets containing 40 ppm feed-grade chlortetracycline. Individual pig average daily gain (ADG), average daily feed intake (ADFI), and gain to feed ratio (G:F) were calculated weekly for 5 weeks. Thereafter, all pigs were euthanized and necropsied for tissue collection. The overall performance data indicated that sCTC pigs had increased ADG (0.43 vs. 0.32 kg/d, P = 0.001) and ADFI (0.51 vs. 0.37 kg/d, P = 0.002) compared with CON pigs; however, G:F was not different as a result of dietary treatment (0.85 vs. 0.88, P = 0.617). Intestinal barrier permeability, ileal active nutrient transport, and cecal short chain fatty acid concentrations did not differ (P > 0.10) due to dietary treatment, however changes in several ileum mRNA transcripts suggest that inflammation may be reduced in sCTC pigs. Further, the changes observed in the proteomes of the ileum, colon, skeletal muscle, and liver suggest that the sub-therapeutic mode of action of AGPs may include post-absorptive changes and warrants further investigation.

Effects of distillers dried grains with solubles on amino acid digestibility, growth performance, and carcass characteristics of growing pigs.

Two experiments were conducted to compare the standardized ileal digestibility (SID) of AA by growing pigs in European distillers dried grains with solubles (DDGS) produced from wheat, maize, or wheat-maize mixtures and to test the effect of increasing the inclusion levels of wheat DDGS on growth performance of growing-finishing pigs fed diets balanced for NE and SID Lys. In Exp. 1, 12 barrows (initial BW: 23.0 ± 2.2 kg) were surgically equipped with a T-cannula in the distal ileum and randomly allotted to a replicated 6 × 6 Latin square design with six diets and six periods. Five sources of European DDGS were used: wheat DDGS from 2011, wheat DDGS from 2012, wheat-80 DDGS (80% wheat and 20% maize), wheat-70 DDGS (70% wheat and 30% maize), and maize DDGS. Each diet contained one source of DDGS as the sole source of AA and an N-free diet was used to determine basal endogenous losses of AA. Results indicated that the SID of CP was greater (P < 0.05) in maize DDGS compared with wheat DDGS from 2011, wheat DDGS from 2012, and wheat-70 DDGS. The SID of all indispensable AA except Trp was also greater (P < 0.05) in maize DDGS compared with all other DDGS sources used in this experiment. For Trp, the SID in wheat-80 DDGS, wheat DDGS from 2011, and wheat DDGS from 2012 were not different from maize DDGS, but were greater (P < 0.05) than in wheat-70 DDGS. The SID for all indispensable AA except Ile and Trp in wheat-70 DDGS were not different from the values calculated for wheat DDGS from 2011 and wheat DDGS from 2012, and no differences between SID values for AA in wheat DDGS from 2011 and wheat DDGS from 2012 were observed. In Exp. 2, 36 growing pigs (initial BW: 38.3 ± 1.97 kg) were randomly allotted to one of four dietary treatments (one pig/pen and nine replicate pigs/treatment) in a 2-phase feeding program (35 to 65, and 35 to 105 kg BW). The four dietary treatments included diets containing 0%, 10%, 20%, or 30% wheat DDGS. Results indicated that there was no effect of wheat DDGS on pig growth performance or carcass quality. However, addition of wheat DDGS increased linearly (P < 0.015) the indole concentration in the carcasses of the pigs. In conclusion, the SID of AA in maize DDGS produced in Europe is greater than in European wheat DDGS and DDGS produced from mixtures of wheat and maize, but inclusion of 30% wheat DDGS in diets fed to growing-finishing pigs did not affect growth performance or carcass quality.

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.

Amino acid digestibility in low-fat distillers dried grains with solubles fed to growing pigs.

The objective of this experiment was to determine the standardized ileal digestibility (SID) of amino acids (AA) in 3 sources of distillers dried grains with solubles (DDGS) with different concentrations of fat. Twelve growing barrows (initial body weight: 76.1 [Formula: see text] 6.2 kg) were randomly allotted to a replicated 6 × 4 Youden square design with 6 diets and 4 periods. The fat content of the 3 sources of DDGS were 11.5, 7.5, and 6.9% respectively. Diets contained 60% DDGS and fat concentration of the diets were 7.5, 5.2, and 5.2%, respectively. Two additional diets containing the 2 sources of DDGS with 7.5 and 6.9% fat were also formulated, and corn oil was added to these diets to increase the concentration of fat in the diets to levels that were calculated to be similar to the diet containing conventional DDGS with 11.5% fat. A N-free diet was also formulated to calculate endogenous losses of crude protein (CP) and AA from the pigs. Pigs were fed experimental diets during four 7-d periods. The first 5 d of each period were an adaptation period and ileal digesta were collected on d 6 and 7 of each period. The apparent ileal digestibililty (AID) and SID of CP and all indispensable AA, except AID Pro and SID of Trp, were greater (P < 0.01) in conventional DDGS than in the 2 sources of DDGS with reduced fat. Adding oil to the diets containing the 2 sources of DDGS with reduced fat did not consistently increase SID of AA. In conclusion, conventional DDGS has greater SID values for most AA compared with DDGS that contains less fat and inclusion of additional oil to diets containing low-fat DDGS does not increase AID or SID of AA. The lower AA digestibility in low-fat DDGS could not be overcome by the inclusion of additional fat to the diets.