Control Pigs Research Articles

Effects of β-glucan on Salmonella enterica serovar Typhimurium swine colonization and microbiota alterations

BackgroundThe 2017 Veterinary Feed Directive eliminated the use of medically important antibiotics for growth promotion of food animals; thus, alternative growth promoters are highly desirable by food animal producers to enhance animal health and reduce pathogen colonization, including the human foodborne pathogen Salmonella. β(1-3)(1-6)-d-glucan (β-glucan) is a soluble fiber with prebiotic characteristics; it has been shown to modulate immune and intestinal functions that strengthen swine resistance to health challenges such as bacterial infections when supplemented in the diets of growing pigs. The current study evaluated the effects of a β-glucan product on gut microbial community structure as well as Salmonella shedding and intestinal colonization.ResultsFive-week-old pigs were fed a β-glucan amended diet at 500 g/ton (n = 13) or a non-amended control diet (n = 14) for three weeks, followed by inoculation of the 27 pigs with 1 × 109 colony forming units of Salmonella enterica serovar Typhimurium strain UK1. While remaining on the respective diets, fecal samples collected at 2, 4, 7, and 16 days post-inoculation (dpi) were similar for Salmonella shedding counts between the two diets. At 16 dpi, Salmonella counts were significantly lower in the cecal contents of the β-glucan-fed pigs (P = 0.0339) and a trend towards a reduction was observed in the Peyer’s patches region of the ileum (P = 0.0790) compared to the control pigs. Pigs fed β-glucan for three weeks exhibited an increase in members of the Clostridia class in their fecal microbial communities, and after inoculation with Salmonella, several potentially beneficial microorganisms were enriched in the microbiota of β-glucan-fed pigs (Lactobacillus, Ruminococcaceae, Prevotellaceae, Veillonellaceae, Bifidobacterium and Olsenella).ConclusionAdministration of β-glucan altered the swine gut microbiome and reduced Salmonella colonization in the cecal contents.

The Effect of Dosing of Stromal Cell-Derived Factor 1 on Anal Sphincter Regeneration.

Aim: The aim of this study is to investigate if a high dose of the stromal cell-derived factor-1 (SDF-1) plasmid improves outcome in a minipig model of chronic anal sphincter injury. Methods: Twenty-two female minipigs underwent excision of the posterior hemicircumference of the anal sphincter complex and were allowed to recover for 6 weeks. They were randomly allocated (n = 6) to receive either 5% dextrose (sham) or 2, 4, or 8 mg of SDF-1 plasmid at the defect site. Two control pigs received no surgery/treatment. Outcome measures included anal manometry at preinjury/pretreatment and 2, 4, and 8 weeks after treatment, recording the mean of eight pressure channels and the posterior channel alone, histopathology using Masson's trichrome, and immunohistochemistry using PGP9.5 for staining of neural structures, and CY3 staining for blood vessels. Data are expressed as mean ± standard error. Manometry analysis used two-way analysis of variance (ANOVA) followed by the Holm-Sidak test. Quantification of muscle/fibrosis was analyzed with a Kruskal-Wallis one-way ANOVA on ranks. Results: Posterior anal pressures were significantly decreased in sham treated animals compared with controls (p = 0.04). In contrast, mean anal pressures at the four time points were not significantly different between groups (p > 0.05). The defect area of the sham treated group showed irregular muscle bundles, while all three SDF-1 treatment groups show organized muscle bundles, with the most organization in the higher dose groups. Quantification of Masson-stained slides showed no statistically significant differences between groups, but did show increased muscle volume in the area of defect in the treatment groups compared with sham. PGP9.5 and CY3 staining showed increased fluorescence in the higher dose groups compared with sham treatment. Conclusion: A single higher dose of the plasmid encoding SDF-1 may increase muscle volume in the area of a chronic defect. Impact statement Fecal or bowel incontinence as a result of a torn anal sphincter complex remains undetected for many years. The resulting defect does not respond well to surgical repair. Regenerating the anal sphincter complex with functional muscle has been a long-term goal. Stem cells home to a site of a chronic injury and cause regeneration when a cell signaling mechanism is available. Stromal cell-derived factor-1 is one such cytokine that has been well researched by us and others to have this effect. It is easy to use clinically and has been used in other applications in humans and considered safe.

Inactivation of growth differentiation factor 9 blocks folliculogenesis in pigs†.

Growth differentiation factor 9 (GDF9) is a secreted protein belonging to the transforming growth factor beta superfamily and has been well characterized for its role during folliculogenesis in the ovary. Although previous studies in mice and sheep have shown that mutations in GDF9 disrupt follicular progression, the exact role of GDF9 in pigs has yet to be elucidated. The objective of this study was to understand the role of GDF9 in ovarian function by rapidly generating GDF9 knockout (GDF9-/-) pigs by using the CRISPR/Cas9 system. Three single-guide RNAs designed to disrupt porcine GDF9 were injected with Cas9 mRNA into zygotes, and blastocyst-stage embryos were transferred into surrogates. One pregnancy was sacrificed on day 100 of gestation to investigate the role of GDF9 during oogenesis. Four female fetuses were recovered with one predicted to be GDF9-/- and the others with in-frame mutations. All four had fully formed oocytes within primordial follicles, confirming that knockout of GDF9 does not disrupt oogenesis. Four GDF9 mutant gilts were generated and were grown past puberty. One gilt was predicted to completely lack functional GDF9 (GDF9-/-), and the gilt never demonstrated standing estrus and had a severely underdeveloped reproductive tract with large ovarian cysts. Further examination revealed that the follicles from the GDF9-/- gilt did not progress past preantral stages, and the uterine vasculature was less extensive than the control pigs. By using the CRISPR/Cas9 system, we demonstrated that GDF9 is a critical growth factor for proper ovarian development and function in pigs.

Effect of intramuscular vaccination against Lawsonia intracellularis on production parameters, diarrhea occurrence, antimicrobial treatment, bacterial shedding, and lean meat percentage in two Danish naturally infected finisher pig herds

Lawsonia intracellularis (LI) is an economically important enteric pathogen in pigs with a worldwide endemic prevalence. The objective of this study was to evaluate the effect of an intramuscularly administrated LI vaccine (Porcilis®Lawsonia Vet.) in Danish finisher pigs (30–115 kg) measured on key production figures, antimicrobial (AB) treatments, occurrence of diarrhea and LI shedding.The study was a group-randomized block-trial with parallel groups in two herds, Herd 1 and Herd 2, experiencing a natural subclinical-clinical LI infection in early finisher period. Vaccination occurred at weaning, but the study focused on the first eight weeks in the finisher period. Further, slaughterhouse data were included.In total, 52 and 50 finisher pens comprising 2184 and 2254 finisher pigs were included in each of two herds, respectively. LI vaccination significantly reduced feed conversion ratio (FCR) by 0.05 and 0.09 FU/kg (p = 0.007 and p < 0.001) alongside a significantly increased average daily weight gain (ADWG) by 31 and 43 gr/day (p = 0.001 and p < 0.001) in each of the herds, respectively. In the vaccinated group, less variation was found in ADWG compared to the control group (p < 0.001 in both herds) as an expression of a more uniform growth, which was further confirmed by less variation in lean meat percent in the vaccinated group in one herd (p = 0.007). No significant difference between groups were found in mortality and pigs excluded due to welfare reasons. AB flock treatment against diarrhea was significantly reduced in Herd 1 with all pens treated in the control group compared to 30.8 % in the vaccinated group (p < 0.001). In Herd 2, the difference was non-significant with 68.0 % in the control group compared to 50.0 % in the vaccination group (p = 0.252). Low levels of individual treatments against diarrhea were seen in both herds (≤ 5.0 %) but still significantly reduced in vaccinated pigs compared to control pigs (p < 0.050 in both herds). Mean diarrheic blot counts were significantly reduced in vaccinated pens compared to control pens (p < 0.001 in both herds). In vaccinated pigs, shedding of LI was reduced in both prevalence (p < 0.001 in both herds), excretion level in positive samples (p < 0.001 in both herds) and, in one herd, also in duration (p = 0.003) when compared to control pigs.In conclusion, pigs vaccinated with Porcilis®Lawsonia Vet against LI in both of two high-health and high-productive finisher herds had, compared to non-vaccinated pigs, significantly improved key production figures, and reduced AB treatment, occurrence of diarrhea, LI shedding, and growth variation.

Dietary embelin supplementation during mid-to-late gestation improves performance and maternal-fetal glucose metabolism of pigs.

This study aimed to evaluate the effects of dietary embelin supplementation during late gestation (from days 60 to 110) on performance and maternal-fetal glucose metabolism of pigs. Sixty sows (Duroc × Yorkshire × Landrace; parity = 1.68 ± 0.03; N = 20) were randomly divided into three gestation (day 60 of pregnancy) treatments, Control pigs (CON) were fed a basal diet, and the other animals were fed a basal diet supplemented with 200 or 600 mg/kg embelin per kg of feed. The body weight, backfat thickness and litter size of the sows, and birth weight and mortality of piglets were recorded. Sows' blood and piglets' umbilical cord blood were collected for the measurements of hematological parameters and anti-oxidative and immune indexes, and maternal-fetal glucose metabolism parameters, respectively. The colostrum and milk and fecal samples of the sows were also collected for analysis of milk composition and apparent total tract nutrient digestibility. Dietary embelin had no effect on the BW and backfat thickness of the sows but significantly increased the birth weight of piglets (P < 0.05) and decreased the mortality (P < 0.05). Moreover, the white blood cell counts (day 90), neutrophil count and mean cell hemoglobin (day 110), total anti-oxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) content of the sows were increased significantly (P < 0.05) in the embelin groups than that in the CON group, whereas the malondialdehyde (MDA) content was decreased (P < 0.05). Embelin significantly increased immunoglobulin A (IgA) and immunoglobulin G (IgG) content in plasma of piglets as well as those in colostrum and milk of sows than the CON treatment (P < 0.05). In addition, dry matter, ash, and ether extract in the colostrum were similar between groups (P > 0.05), whereas the embelin significantly increased the crude protein in the milk. The apparent total tract nutrient digestibility was similar between treatments (P > 0.05). The embelin treatment significantly increased the glucose levels and lactate dehydrogenase B (LDHB) activity in sows plasma, and decreased the lactate levels in both sows and fetuses plasma (P < 0.05). Collectively, this study indicates that sows fed with embelin in mid-to-late gestation showed improved maternal health and anti-oxidative status, milk protein content, and maternal-fetal glucose metabolism, showing promise in natural plant extract nutrition for sows.

A second iron injection administered to piglets during lactation improves hemoglobin concentration, growth performance, and carcass characteristics at slaughter.

An experiment was conducted to evaluate the effects of a second injection of iron dextran administered on days 6 to 8 of age. A total of 144 crossbred pigs (equal barrows and gilts; initial age 6 to 8 d; initial body weight [BW] = 2.86 ± 0.01 kg) were assigned to either the control (CON) or an added-injection treatment (+Fe). Pigs were paired by sex and BW within a litter and randomly assigned to the iron treatment within each pair. All pigs had received an initial intramuscular (IM) injection of iron dextran (200 mg Fe) <24 h after birth. Pigs assigned to the +Fe treatment received a second IM injection of iron dextran (200 mg Fe) on days 6 to 8. All pigs were weaned at 22 to 25 d, housed 6 pigs/pen, and received a common corn-soybean meal diet. BW and feed disappearance were recorded every 2 wk. Hemoglobin (Hb) concentrations were measured at birth, initiation of experiment (days 6 to 8), weaning, and the end of the nursery and end of the study. At the end of the study, 1 pig/pen (n = 12 pigs/treatment), closest to the pen mean was selected and slaughtered for carcass characteristic measures. The individual pig served as the experimental unit for BW, Hb, average daily gain (ADG), and carcass characteristic data whereas the pen served as the experimental unit for average daily feed intake, and gain/feed ratio data. The +Fe pigs had a greater Hb at weaning (13.1 vs. 10.7 g/dL, respectively; P < 0.01) and end of the nursery (12.1 vs. 11.7 g/dL, respectively; P = 0.01) compared to CON pigs. During the finisher period, +Fe pigs had a greater ADG (0.94 vs. 0.91 kg, respectively; P = 0.05) compared to CON pigs. Overall, pigs receiving the second iron injection had an ~4% increase in ADG (P = 0.04) from weaning to the end of study. The cumulative improvement in ADG from weaning to the end of study observed for +Fe group resulted in +Fe pigs having a heavier BW at the end of the study (~3 kg; P = 0.04). Following slaughter, +Fe pigs had ~7.2% heavier trimmed loin (P = 0.04) compared to the CON pigs. In conclusion, administering a second iron injection resulted in greater Hb at weaning and the end of the nursery as well as improved growth performance from weaning to the end of study weight and increased carcass weight at slaughter.

Compensatory gain based on lysine level in finishing pigs after being fed lysine deficient 97% corn diets for 3 or 6 wk.

The objective of this experiment was to evaluate increasing the concentration of lysine on the compensatory gain of finishing pigs during their recovery period after being fed a 97% corn holding diet for 3 or 6 wk. One thousand six hundred and eighty pigs with a starting body weight of 73.5 ± 2.2 kg were blocked by starting body weight and assigned to a nested arrangement. Twenty replicates of seven treatments were comprised of two restriction lengths [3 weeks (3 wk) vs. 6 weeks (6 wk)], and three lysine concentrations during recovery (Lys:ME same as control:100; control + 10%: 110; control + 20%: 120) plus one control (CONT) that remained nutrient unrestricted. Pen weight and feed intake were recorded on days 0, 21, 41, and at marketing. Whole pens were marketed when the pen average met 130 kg and carcass measurements were collected. Data were analyzed by pen with the fixed effects of restriction length and lysine within restriction length. Nutrient restriction lowered (P < 0.01) average daily gain (ADG) compared to control, with 1.2, 0.4, and 0.5 kg for control, 3 wk, and 6 wk treatments, respectively. Restricted pigs showed decreased feed intake while restricted. After the respective restriction period, pigs were allowed a recovery diet until market. Previously restricted pigs had 16.7% and 27.3% greater (P < 0.01) ADG over control pigs for 3 and 6 wk treatments, respectively, in the first 3-wk of recovery. The lysine concentration in the recovery diet impacted (P < 0.01) the ADG with pigs allowed the highest lysine concentration having a 10% greater ADG than pigs fed the lower Lys:ME concentrations, for both restriction treatments. The increase in ADG was not paralleled by an increase in feed intake over control, thus, there was an improvement (P < 0.01) in gain to feed ratio in the recovery period. Control pigs reached market weight (131.5 kg) on experiment day 49 while pigs fed corn diets for 3 wk or 6 wk were slower to market (57 and 69 days, respectively; P < 0.01). Restricted pigs had greater backfat (CONT: 1.47, 3 wk: 1.55, 6 wk: 1.65 cm; P < 0.01), and decreased loin depth (CONT: 7.32, 3 wk: 7.03, 6 wk: 6.61 cm, P < 0.02) which was also impacted (P < 0.01) by lysine concentration. In conclusion, the use of restrictive diets reduced ADG and increased days to market. The use of recovery diets in which the Lys:ME ratio was greater than control pigs, resulted in increased compensatory growth.

Metabolic insights and background from naturally affected pigs during Streptococcus suis outbreaks.

Streptococcus suis (S. suis) is an endemic zoonotic pathogen still lacking adequate prevention in pigs. The present case study looked back to the occurrence and consequences of S. suis outbreaks in our swine research facilities in search of new metabolic and physiological insight. From a series of outbreaks, a dataset was created including 56 pigs sampled during disease detection based on clinical signs. Pigs suspected with S. suis infection were defined as diseased (n = 28) and included pigs defined as neurologically diseased (n = 20) when severe neurological signs (central nervous system dysfunctions, i.e., opisthotonos, ataxia, and generalized tremor) were observed. Another set of 28 pigs included respective pen mates from each case and were defined as control. Representative deaths were confirmed to be caused by S. suis. Tonsillar swabs were collected and analyzed by quantitative polymerase chain reaction (qPCR) for total bacteria, total S. suis, and S. suis serotypes (SS) 2 (and/or 1/2) and 9. Blood and sera were analyzed to quantify blood gases, minerals, and S. suis reactive immunoglobulins against current isolates. Data collected included litter sibling associations, birth and weaning body weight (BW), and average daily gain (ADG) 7 d after the disease detection. In general, the disease increased pH, sO2 and the incidence of alkalosis, but reduced pCO2, glucose, Ca, P, Mg, K, and Na in blood/serum compared to control. The SS2 (and/or SS1/2) prevalence was significantly (P < 0.05) increased in neurologically diseased pigs and its relative abundance tended (P < 0.10) to increase in tonsils. In contrast, the relative abundance of total S. suis was lower (P > 0.05) in diseased pigs than control pigs. Levels of S. suis reactive IgG2 were lower, but IgM were higher (P < 0.03) in neurologically affected pigs compared to control. Furthermore, there was an increased proportion of sibling pigs that were diseased compared to control. In conclusion, our results evidence that naturally affected pigs were associated to average performing pigs without any predisease trait to highlight but a sow/litter effect. Besides, neurologically affected pigs had increased S. suis (SS2 and/or 1/2) prevalence and relative abundance, a respiratory alkalosis profile, and mineral loss.

МИКРОБИОМ КИШЕЧНИКА ПОРОСЯТ В ПЕРИОД ДОРАЩИВАНИЯ ПРИ ИСПОЛЬЗОВАНИИ ПРОБИОТИКА «ЛИКВАФИД»

The article presents the results of a study of the effect of the LiquaFid probiotic on the colonic microbiome of pigs during the growing period. When using the real-time PCR method, it was found that the probiotic complex contributed to an increase in the total number of bacteria in the contents of the colon of the intestine relative to the group of control pigs. In experimental piglets, an increase in the proportion of normoflora representatives in the bacterial community is noted. In the structure of the normoflora of the colon calving in experimental animals, there is a quantitative increase in representatives of the genera Prevotella spp., Porphyromonas spp., Eubacterium spp., Lachnobacterium spp., Clostridium spp., Megasphaera spp., Veillonella spp., Dialister spp. and Lactobacillus spp. Against the background of the growth of representatives of the normoflora, a decrease in opportunistic and pathogenic microorganisms is observed.

Effects of low-protein diet and feed restriction on mRNA expression of cationic amino acid transporters in porcine skeletal muscles.

We investigated the effects of a low-protein diet and feed restriction on the mRNA expression of cationic amino acid transporters (CATs) in the longissimus dorsi (LD), rhomboideus (RH), and biceps femoris (BF) muscles of pigs. Eighteen piglets were divided into three groups: a control (CP21%), low-protein diet (LP, CP16%), and feed-restricted diet (FR, CP21%, 76% feed intake of control pigs) groups. The expression levels of CAT-1 in the LD and BF muscles of LP pigs were higher than that of control pigs, whereas that of FR pigs showed no difference. The CAT-2A expression levels in the RH muscle of FR pigs were higher than that of control pigs. The free lysine concentrations in all muscles of LP and FR pigs were lower than that of control pigs. To examine the factors that affect CATs mRNA expression, we evaluated the effects of lysine, arginine, insulin-like growth factor-I, and dexamethasone on the expression of CATs in C2C12 myotubes. CAT-1 expression levels increased in lysine and/or arginine deprivation. We show that CAT-1 and CAT-2A expression levels in skeletal muscles differ in response to dietary treatments and CAT-1 expression in skeletal muscles appears to increase in response to low free lysine concentrations.

H2O2 Mediates VEGF- and Flow-Induced Dilations of Coronary Arterioles in Early Type 1 Diabetes: Role of Vascular Arginase and PI3K-Linked eNOS Uncoupling

In diabetes, the enzyme arginase is upregulated, which may compete with endothelial nitric oxide (NO) synthase (eNOS) for their common substrate L-arginine and compromise NO-mediated vasodilation. However, this eNOS uncoupling can lead to superoxide production and possibly vasodilator hydrogen peroxide (H2O2) formation to compensate for NO deficiency. This hypothesis was tested in coronary arterioles isolated from pigs with 2-week diabetes after streptozocin injection. The NO-mediated vasodilation induced by flow and VEGF was abolished by NOS inhibitor L-NAME and phosphoinositide 3-kinase (PI3K) inhibitor wortmannin but was not affected by arginase inhibitor Nω-hydroxy-nor-L-arginine (nor-NOHA) or H2O2 scavenger catalase in control pigs. With diabetes, this vasodilation was partially blunted, and the remaining vasodilation was abolished by catalase and wortmannin. Administration of L-arginine or nor-NOHA restored flow-induced vasodilation in an L-NAME sensitive manner. Diabetes did not alter vascular superoxide dismutase 1, catalase, and glutathione peroxidase mRNA levels. This study demonstrates that endothelium-dependent NO-mediated coronary arteriolar dilation is partially compromised in early type 1 diabetes by reducing eNOS substrate L-arginine via arginase activation. It appears that upregulated arginase contributes to endothelial NO deficiency in early diabetes, but production of H2O2 during PI3K-linked eNOS uncoupling likely compensates for and masks this disturbance.

Attenuation of streptozotocin induced high fat diet exacerbated dyslipidemia mediated hepatic and aortic injuries in male pigs by camel milk

ABSTRACT Dyslipidemia, diabetes, and the upregulation of receptors for oxidized low-density lipoprotein all contribute to hepatic and aortic endothelial dysfunction. The current research examined how camel milk could possibly decrease induced Streptozotocin-high fat diet-aggravated dyslipidemia-mediated damage in male pig hepatic and aortic tissues. Twenty-five pigs were separated into five groups of five pigs each, with five non-atherosclerotic pigs in the control group and twenty atherosclerotic pigs in the experimental groups. Groups 1 and 2 received distilled water as the standard control and atherosclerotic control groups, respectively, while Groups 3 and 4 received camel milk at 250 mL/day and 500 mL/day, respectively, and Group 5 received Metformin at 500 mg/day. The experiment lasted ten weeks. After 10 weeks, all of the pigs were euthanized. The lipid profile and liver enzyme levels were determined from the analysis of blood serum and the histology of the liver and aortic. When pigs administered, camel milk were compared to atherosclerotic control pigs, there was a significant decrease (p 0.05) in cholesterol, triglyceride, low density lipoprotein (LDL), and very low-density lipoprotein (VLDL) levels, while a significant increase in high density lipoprotein levels was observed in pigs that received camel milk. Also, pigs in groups that received camel milk had their liver enzyme parameters (ALP, AST, ALT, GGT, LDH, and CK-MB) significantly lowered when compared with atherosclerotic control pigs. Similarly, treatment with camel milk ameliorates aortic tissues from degradation and reduces histological liver steatosis, and it also decreases the expression of LOX-1 mRNA in the aorta of atherosclerotic pigs.

Feasibility of a new bariatric fully endoscopic duodenal-jejunal bypass: a pilot study in adult obese pigs

This study assessed a new natural orifice transluminal endoscopic surgery (NOTES) bariatric endoscopic procedure in obese adult pigs. This 14-week prospective study compared 6 adult male Yucatan test pigs with baseline mean age 1.5 years to 2 control pigs. The test pigs received a fully endoscopic NOTES-based duodenal-jejunal bypass including measurement of the bypassed limb and creation of a gastrojejunal anastomosis (GJA) using a gastrojejunal lumen-apposing metal stent (GJ-LAMS) at Week 0, placement of a duodenal exclusion device (DED) at Week 2, and endoscopic examinations at Weeks 6 and 10. At Week 14, the pigs were sacrificed for necropsy. All endoscopic procedures were technically successful. By Week 14, the controls had gained a mean 1.1 ± 2.1 kg (+ 1.6%) while the intervention animals lost a mean 6.8 ± 3.9 kg (− 10.5%) since baseline. GJ-LAMS migrations occurred in 3 pigs, two of which also had DED migration and/or partial dislocation. Two other pigs died, one with aberrant splenic vein positioning near the GJA and the other from an unknown cause. An endoscopic bariatric bypass procedure with controlled bypass length was technically successful in all the cases and led to weight loss in test animals. Safety concerns must be further addressed.

Identification of novel antisense long non-coding RNA APMAP-AS that modulates porcine adipogenic differentiation and inflammatory responses

Long non-coding RNAs (lncRNAs) are emerging as powerful regulators of adipocyte differentiation, fat metabolism and gene expression. However, the functional roles and mechanisms of lncRNAs in these processes remain unclear. Here, we identified a novel antisense transcript, named APMAP-AS, transcribed from adipocyte membrane-associated protein (APMAP) in the pig genome. APMAP-AS and APMAP were highly expressed in retroperitoneal adipose of obese pigs, compared with that in control pigs. Using a bone mesenchymal stem cells (BMSCs) adipogenic differentiation model, we found that APMAP-AS positively regulated adipogenic differentiation. APMAP-AS had the potential to form an RNA–RNA duplex with APMAP, and increased the stability of APMAP mRNA. Additionally, APMAP-AS promoted lipid metabolism and inhibited the expression of inflammatory factors. These findings of a natural antisense transcript for a regulatory gene associated with lipid synthesis might further our understanding of lncRNAs in driving adaptive adipose tissue remodeling and preserving metabolic health.

Mechanical Ventilation in ARDS With an Automatic Resuscitator.

The Oxylator is an automatic resuscitator, powered only by an oxygen cylinder with no electricity required, that could be used in acute respiratory failure in situations in which standard mechanical ventilation is not available or feasible. We aimed to assess the feasibility and safety of mechanical ventilation by using this automatic resuscitator in an animal model of ARDS. A randomized experimental study in a porcine ARDS model with 12 pigs randomized to the Oxylator group or the control group (6 per group) and ventilated for 4 h. In the Oxylator group, peak pressure was set at 20 cm H2O and PEEP was set at the lowest observed breathing frequency during a decremental PEEP titration. The control pigs were ventilated with a conventional ventilator by using protective settings and PEEP at the crossing point of collapse and overdistention, as indicated by electrical impedance tomography. Our end points were feasibility and safety as well as respiratory mechanics, gas exchange, and hemodynamics. After lung injury, the mean ± SD respiratory system compliance and PaO2 /FIO2 were 13 ± 2 mL/cm H2O and 61 ± 17 mm Hg, respectively. The mean ± SD total PEEP was 10 ± 2 cm H2O and 13 ± 2 cm H2O in the control and Oxylator groups, respectively (P = .046). The mean plateau pressure was kept to < 30 cm H2O in both groups. In the Oxylator group, the tidal volume was transiently > 8 mL/kg but was 6 ± 0.4 mL/kg at 4 h, whereas the breathing frequency increased from 38 ± 4 to 48 ± 3 breaths/min (P < .001). There was no difference in driving pressure, compliance, PaO2 /FIO2 , and pulmonary shunt between the groups. The mean ± SD PaCO2 was higher in the Oxylator group after 4 h, 74 ± 9 mm Hg versus 58 ± 6 mm Hg (P < .001). There were no differences in hemodynamics between the groups, including blood pressure and cardiac output. Short-term mechanical ventilation by using an automatic resuscitator and a fixed pressure setting in an ARDS animal model was feasible and safe.

Cardiac epigenetic changes in VEGF signaling genes associate with myocardial microvascular rarefaction in experimental chronic kidney disease.

Chronic kidney disease (CKD) is common in patients with heart failure and often results in left ventricular diastolic dysfunction (LVDD). However, the mechanisms responsible for cardiac damage in CKD-LVDD remain to be elucidated. Epigenetic alterations may impose long-lasting effects on cellular transcription and function, but their exact role in CKD-LVDD is unknown. We investigate whether changes in cardiac site-specific DNA methylation profiles might be implicated in cardiac abnormalities in CKD-LVDD. CKD-LVDD and normal control pigs (n = 6 each) were studied for 14 wk. Renal and cardiac hemodynamics were quantified by multidetector CT and echocardiography. In randomly selected pigs (n = 3/group), cardiac site-specific 5-methylcytosine (5mC) immunoprecipitation (MeDIP)- and mRNA-sequencing (seq) were performed, followed by integrated (MeDiP-seq/mRNA-seq analysis), and confirmatory ex vivo studies. MeDIP-seq analysis revealed 261 genes with higher (fold change > 1.4; P < 0.05) and 162 genes with lower (fold change < 0.7; P < 0.05) 5mC levels in CKD-LVDD versus normal pigs, which were primarily implicated in vascular endothelial growth factor (VEGF)-related signaling and angiogenesis. Integrated MeDiP-seq/mRNA-seq analysis identified a select group of VEGF-related genes in which 5mC levels were higher, but mRNA expression was lower in CKD-LVDD versus normal pigs. Cardiac VEGF signaling gene and VEGF protein expression were blunted in CKD-LVDD compared with controls and were associated with decreased subendocardial microvascular density. Cardiac epigenetic changes in VEGF-related genes are associated with impaired angiogenesis and cardiac microvascular rarefaction in swine CKD-LVDD. These observations may assist in developing novel therapies to ameliorate cardiac damage in CKD-LVDD.NEW & NOTEWORTHY Chronic kidney disease (CKD) often leads to left ventricular diastolic dysfunction (LVDD) and heart failure. Using a novel translational swine model of CKD-LVDD, we characterize the cardiac epigenetic landscape, identifying site-specific 5-methylcytosine changes in vascular endothelial growth factor (VEGF)-related genes associated with impaired angiogenesis and cardiac microvascular rarefaction. These observations shed light on the mechanisms of cardiac microvascular damage in CKD-LVDD and may assist in developing novel therapies for these patients.

Abstract 11537: IGF-1 Reduces Atherosclerosis in Rapacz Pigs

Introduction: Cardiovascular disease, including atherosclerosis is the leading cause of mortality in the Western world. We have reported that insulin-like growth factor I (IGF-1) reduced atherosclerosis in the murine model, however it is unknown whether IGF-1 exerts protective effect on human-like atherosclerosis. Hypothesis: Rapacz pigs with familial hypercholesterolemia (FH pigs) fed with high-cholesterol diet develop human-like advanced plaques in coronary arteries. We hypothesize that IGF-1 suppresses atherosclerosis in FH pigs. Methods: Human recombinant IGF-1 ( 50 ug/kg) or saline (control) was injected daily into high-fat diet fed FH swine (n=9/group) for 6 months. Atherosclerotic burden was quantified by intravascular ultrasound (IVUS) at basal level (T0), after 3 months on diet (T3) and at sacrificing (T6). Results: IGF-1-injected pigs had 2.6-fold increase in total plasma IGF-1 (ELISA, p&lt;0.001 vs. control pigs). Pigs in IGF-1 and control groups had a gradual increase in vessel volume in the right coronary artery (RCA), and left anterior descending artery (LAD) with time as quantified by IVUS. However, IGF-1-injected pigs had a larger increase in vessel volume after 3 and 6 months compared to controls suggesting vascular hypertrophy. The atherosclerotic burden was increased in RCA, and LAD in both groups after 3 and 6 months of injections compared to the pre-injection time point. We found a significant reduction in atherosclerotic burden in RCA and LAD in IGF-1 group compared to control at 6 months’ time-point (RCA: IGF-1, 203±13%, vs. control, 244±8; LAD: IGF-1, 200±6%, vs. control, 228±11%). The lumen volume of coronaries was decreased in both groups, however, coronaries in IGF-1-injected pigs had on average 24±3% larger lumen volume compared to control pigs at 6 months’ time-point. Conclusions: We found that IGF-1 increased total volume of pig coronaries suggesting hypertrophy and these data are consistent with growth-stimulating effects of IGF-1. We found reduction in atherosclerotic burden and increase in lumen volume in coronaries in the IGF-1 group after 6 months of injections consistent with an anti-atherosclerotic effect of IGF-1. Our data support the feasibility of using IGF-1 for as an anti-atherogenic therapy.

Olfactory enrichment using a maternal pheromone improved post-weaning pig performance and behavior.

The post-weaning environment in commercial pig nursery buildings is typically devoid of both the sow and her semiochemicals. Among other factors, the loss of maternal odors may contribute to post-weaning stress. In this work, we report four studies (studies 1-4) using 192 weaned pigs to evaluate the effects of a maternal pheromone (MP) containing Myristic Acid and Skatole on weaned pig behavior and performance. Study 1 examined behavior among weaned pigs with a focus on lying, aggression and feeding behaviors. All studies used body weight gain and the percentage of pigs that lost weight the first 7 days after weaning as key response variables. The MP stimulated early feeding behavior and reduced aggressive behaviors among weaned pigs (p < 0.01). In study one, pigs were over 4 kg heavier 16 weeks post-weaning when the MP was present for 48 h post-weaning compared to control pigs (p = 0.05). The three other studies used a 7-days bioassay to assess the best dose and form of delivery of the MP among weaned pigs. The single measure that responded uniformly was the percentage of pigs that lost body weight in the first 7 days after weaning. Among control pigs in four separate studies, 25% of the weaned pigs in each study lost body weight by 7 days after weaning, while in 3 of the 4 bioassay studies, 0% of MP-exposed pigs lost weight after weaning (one study had 6% of pigs losing body weight with the MP). This MP effect is both highly statistically significant and, if found on commercial farms, would have a large positive economic benefit. Simultaneously, having fewer pigs losing weight and exhibiting less aggressive behavior will improve the health and welfare of weaned pigs. Providing biologically-relevant olfactory enrichment can improve pig health and welfare.

Application of a β -Mannanase Enzyme in Diets with a Reduced Net Energy Content Results in Reduced Production Costs Per Kg of Carcass Weight

β-Mannans are strongly anti-nutritive polysaccharide fibres found in most vegetable feed ingredients. They belong to the hemicellulose fraction and have a backbone composed entirely of mannose, as in mannans and galactomannans, or of mannose and glucose, as in glucomannans and galacto-glucomannans. The estimated content of soluble β-mannans in common fattening diets is only 0.20-0.35%, and in vitro studies have demonstrated that as little as 0.05% soluble β-mannan in feed can elicit a strong innate immune response. This innate response is often referred to as a feed induced immune response or FIIR, which suppresses growth to protect the liver and reserve energy and nutrients for high priority immune functions. Hemicell HT (Elanco Animal Health) is a β-mannanase enzyme for animal feed that breaks down β-mannans and thereby prevents economic losses from the wasteful immune response to β-mannans. The aim of the present study was to compare pig performance and carcass parameters on a control diet and a reformulated diet with lower energy content - a 65 kcal reduction in net energy per kg – and inclusion of a β-mannanase enzyme. An eighteen-week feeding trial was conducted on a commercial fattening unit with DanBred x Belgian Piétrain pigs starting at 10 weeks of age. Standard two-phase control diets (Control group) were compared to reformulated diets with an energy reduction of 65 kcal NE/kg and inclusion of a β-mannanase enzyme (Hemicell HT; Elanco) at 300 g/tonne (Enzyme treated group). Standard production and health data were collected including days in fattening, bodyweight at start, at 28 days and at 127 days (prior to slaughter), feed intake, mortality and antibiotic use. Additionally, Average Daily Weight Gain (ADWG), Average Daily Feed Intake (ADFI) and Feed Conversion Rate (FCR) were calculated from the collected data. Furthermore, pigs were slaughtered on separate slaughter days to reliably collect relevant carcass parameters at slaughterhouse level. The data were analysed using JMP 15.0 statistical program. Overall, performance data did not differ significantly between trial groups in both Phase 1 and Phase 2, except for mortality that was significantly higher during Phase 1 in the Control group (3.19% vs. 0.00% in the Enzyme treated group). Carcass quality did only significantly (P &lt; 0.05) differ for muscle depth (73.58 ± 0.66 mm vs. 75.33 ± 0.59 mm in the Control and Enzyme treated group, respectively). Following calculation of feed costs per kg carcass weight and considering the cost of enzyme inclusion, Enzyme treated pigs had € 0.033 lower feed costs per kg carcass weight as compared to Control pigs. The trial demonstrated that inclusion of Hemicell HT in reformulated diets with a lower energy content (65 kcal NE/kg) was able to degrade β-mannans in diets and therefore maintain production performance and overall carcass quality. Reduction in mortality may indicate an improved overall immunity due to decreased FIIR by the breakdown of β-mannans in the feed. Inclusion of a β-mannanase enzyme resulted in an overall reduction in production costs of € 0.033 per kg of carcass weight, which is an additional advantage considering the current feed prices globally.

An oral gavage of lysine elicited early satiation while gavages of lysine, leucine, or isoleucine prolonged satiety in pigs.

Excess dietary amino acids (AA) may negatively affect feed intake in pigs. Previous results showed that Lys, Leu, Ile, Phe, and Glu significantly increased gut peptide secretion (i.e., cholecystokinin, glucagon-like peptide 1). However, the link between dietary AA and gut peptide secretion with changes in feeding behavior patterns has not been demonstrated to date in pigs. The aim of the present study was to determine the effect of Lys, Leu, Ile, Phe, and Glu, on feed intake and meal patterns in young pigs. Twelve male pigs (Landrace × Large White, body weight = 16.10 ± 2.69 kg) were administered an oral gavage of water (control) or Lys, Leu, Ile, Phe, Glu, or glucose (positive control) at 3 mmol.kg-1 following an overnight fasting. The experiment consisted in measuring individual feed disappearance and changes in meal pattern (including latency to first meal, first meal duration, intermeal interval, second meal duration, and number of meals) based on video footage. Compared to the control group Lys significantly (P ≤ 0.01) reduced feed intake during the first 30 min and up to 2.5 h post-gavage, including a reduction (P ≤ 0.05) in the first meal duration. Similarly, Leu and Ile also significantly decreased feed intake up to 3 h post-gavage on a cumulative count. However, the strongest (P ≤ 0.01) impacts on feed intake by the two branched chained AA were observed after the first- or second-hour post-gavage for Leu or Ile, respectively. In addition, Leu or Ile did not affect the first meal duration (P ≥ 0.05). Leu significantly increased (P ≤ 0.01) the intermeal interval while decreasing (P ≤ 0.05) the number of meals during the initial 2 h following the gavage when compared with the control group. In contrast, the oral gavages of Phe or Glu had no significant impact (P > 0.05) on the feeding behavior parameters measured relative to the control pigs. In turn, glucose had a short-lived effect on appetite by reducing (P < 0.05) feed intake for 30 min after the first-hour post-gavage. In conclusion, the impact of an oral gavage of Lys on feeding behavior is compatible with a stimulation of early satiation and an increased duration of satiety. The main impact of the oral gavages of Leu and Ile was an increase in the duration of satiety. The gastrointestinal mechanisms associated with non-bound dietary AA sensing and the impact on voluntary feed intake warrant further investigations.