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The rumen is a critical organ mediating nutrient uptake and use in cattle. Healthy rumen development is essential to ensure animal feed efficiency. In this work, we present an analysis of transcriptomic dynamics in rumen epithelium during the transition from pre-rumination to rumination in cattle-fed hay or concentrated diets at weaning (eighteen Holstein bull calves, 3 X 6 groups). These two distinct phases of rumen development and function in cattle are tightly regulated by a series of signaling events and clusters of effectors on key pathways. Our analysis identifies putative signaling events and effectors. Gene activity shifts indicated the transcriptomic reprogramming required to induce developmental changes in ruminal epithelium and functional transitions. A principal component analysis distinguished the temporal expression patterns that clustered separately between pre- and post-weaning groups. A GO-term enrichment analysis reflected functional (physical and metabolic) development of ruminal epithelium and revealed the greatest number of DEGs were enriched in biological processes related to energy metabolism. Canonical pathway and upstream regulator analyses revealed transcription reprogramming with clusters of critical pathways and upstream regulators controlling functional and developmental transitions with no significant differences between hay- and concentrate-fed groups at weaning. The most highly activated transcription factors expressed during the weaning transition were PPARGC1A, INSR, NFE2L2, MYC, MYCN, and PPARA. Overall, the dietary shift from liquid to solid feeds prompted transcriptional reprogramming in rumen epithelial tissue reflecting critical nutrient-gene interactions occurring during the developmental progression of ruminant digestion.

The importance of the microbiota in the gastrointestinal tract of animals is recognized as a critical player in host health. Recently, the significance of the mycobiome has been recognized, but culture-independent studies are limited, especially in swine. Weaning is a time of stress, dietary changes, and a predisposition to infections, making it a time point of interest to industry. In this pilot study, we sought to assess and characterize the mycobiome in the feces of swine from birth through the critical weaning transition to investigate the mycobiome population and its temporal dynamics in piglet feces. Cultured fecal samples demonstrate a significant increase in fungal burden following weaning that does not differ from adult levels, suggesting stable colonization. Culturable fungi were not found in any environmental samples tested, including water, food, sow milk or colostrum. To determine the fungal diversity present and to address the problem of unculturable fungi, we performed a pilot study utilizing ITS and 16S rRNA focused primers for high-throughput sequencing of fungal and bacterial species, respectively. Bacterial populations increase in diversity over the experimental timeline (days 1 to 35 postbirth), but the fungal populations do not demonstrate the same temporal trend. Following weaning, there is a dynamic shift in the feces to a Saccharomycetaceae-dominated population. The shift in fungal population was because of the dominance of Kazachstania slooffiae, a poorly characterized colonizer of animal gastrointestinal tracts. This study provides insights into the early colonization and subsequent establishment of fungi during the weaning transition in piglets. Future studies will investigate the effect of the mycobiome on piglet growth and health during the weaning transition.

Increased plasma α-1 acid glycoprotein (AGP) is correlated with reduced growth rates in neonatal swine. The specific physiological mechanisms contributing to this relationship are unknown. This study was performed to determine if AGP can modify muscle metabolism by examining glucose oxidation and protein synthesis in the C2C12 muscle cell line. Cells were used for experiments 4 days post-fusion as myotubes. Myotubes were exposed to AGP for 24 h, with the last 4 h used to monitor 14C-glucose oxidation or to measure protein synthesis by incorporation of 3H-tyrosine. Treatment of C2C12 myotubes with mouse AGP (100 µg/ml) reduced glucose oxidation (P0.05, n=6 trials), whereas incubation with both AGP and insulin reduced 3H-tyrosine release by 15% (P

Embryo transfer is an advanced technology and has great potential in increasing desired livestock without physical boundaries. Multiple embryos can be produced in-vitro or in-vivo using semen from desired males. Early stage embryos (zygotes) are also ideal tools for genome-editing to alter the desired traits. However, the small number of mature oocytes produced each cycle is a major limitation of this technology. Scientists have used drugs to stimulate follicles to increase the number of oocytes/embryos. Extra oocytes/embryos produced can be cryopreserved or shipped to distant locations, to disseminate genetics around the world, at a fraction of cost. Embryos produced from superovulated animals can be transferred in recipient does, to produce desired number of animals. The goal of this study was to estimate the rate of ovulation and oocyte recovery after drug induced superovulation in goats. Eight cycling does were synchronized for estrus, using intravaginal CIDR dispensers, for 11 days during natural breeding season. Each doe was superovulated with a total of 200 mg of pFSH administered intramuscularly in 6 dosages (50 mg, 30 mg, and 20 mg given twice a day at 12 h interval) starting 48 h prior to CIDR removal. Each doe also received 8 mg of PGF2α, 24 h prior to CIDR removal, and 86 µg of GnRH 24 h after CIDR removal. The superovulatory response was evaluated by counting corpus luteum (CL) on each ovary after 48 h of GnRH injection. The ovulated oocytes were recovered surgically from oviducts by flushing with TL Hepes medium. Mean and standard deviation of CL calculated was 10.86±6.34 and 11.67±6.15 CL per doe (n = 8) in left and right ovaries, respectively. Similarly, mean and SD of mature oocytes recovered was 7.67±6.50 and 9.67±5.10 for left and right oviducts, respectively. Average percent recovery of oocytes was 71.23 % / doe.

The importance of the microbiota in the gastrointestinal (GI) tract of animals is recognized as a critical player in host health. Recently, the significance of the mycobiome has been recognized, but culture-independent studies are limited, especially in swine. Weaning is a time of stress, dietary changes, and a predisposition to infections, making it a time of interest to industry. In this study, we sought to assess and characterize the mycobiome and microbiome in the feces and GI tract of swine from birth through the critical weaning transition (days 1–35 post-birth). In addition, we investigated environmental factors that may alter microorganisms present in piglets. Fecal bacterial populations increased in diversity over the experimental timeline and demonstrated a transition from an Enterobacteriaceae-dominated population, to a Prevotellaceae and Ruminococcaceae-dominated population by days 24–35 post-birth. These later populations are capable of fiber degradation and short chain fatty acid production. In fecal fungal populations, richness and diversity peaked at weaning and declined post-weaning. There was also a dynamic shift in the mycobiome to a Saccharomycetaceae-dominated population that remained stable into adulthood. Fungal organisms contributing to this colonization were not found in environmental samples including water, colostrum, and feed. Despite fungal populations present in the feces of sows, these maternal fungi were not similar to the piglet mycobiome and thus did not indicate a maternally-derived effect. Furthermore, the microbiomes of the GI tract showed decreased richness and diversity in the upper GI compared to the lower GI, and a high degree of individual variation and litter effect throughout the organs. This study provides insights into the early colonization and establishment of fungi during the weaning transition. Future studies will investigate the effect of the mycobiome on piglet growth and health during the weaning transition, including their role in fast- versus slow-growing piglets.

Background Recent increases in intra-litter variability in weaning weight have raised swine production costs. A contributor to this variability is the normal birth weight pig that grows at a slower rate than littermates of similar birth weight. The goal of this study was to interrogate biochemical profiles manifested in skeletal muscle originating from slow growing (SG) and faster growing littermates (control), with the aim of identifying differences in metabolic pathway utilization between skeletal muscle of the SG pig relative to its littermates. Samples of longissimus muscle from littermate pairs of pigs were collected at 21 d of age for metabolomic analysis (Metabolon, Inc., Durham, NC). Results Birth weights did not differ between littermate pairs of SG and Control pigs (P > 0.05). Weaning weights differed by 1.51 ± 0.19 kg (P

Porcine α-1 acid glycoprotein (AGP) in newborn pigs can be used to predict growth rate through weaning and is a marker for growth impairment. This study examined whether nutritional support can improve the growth rate of piglets identified as having poor growth potential. Cross-fostering (CF) and CF plus a milk supplement (CF + MS) were used to attempt to improve the growth performance of pigs. Blood was collected at d1 post-parturition for measurement of plasma AGP for all pigs in 28 litters contributing to the experiment. Piglets with the highest plasma AGP level were weight and sex matched to a littermate with a low plasma AGP concentration and four pairs of these weight and sex matched pigs were grouped into four foster litters per treatment (control, CF, CF + MS). The control group was assembled by pairing littermates remaining in donor litters. Pigs stayed on treatment until weaning at 21 days of age. At 35 days of age, dual energy X-ray absorptiometry (DXA) was performed on CF and CF + MS pigs to evaluate carcass composition. Control pairs differed in weaning weight, with pigs with higher plasma AGP at 1 day of age having smaller weaning weights than their littermates of similar birth weight (P + MS produced a similar effect as CF (P > 0.05). At 35 days of age, body weights were still similar between CF littermates and between CF + MS littermates (P > 0.05). DXA analysis demonstrated that body composition was similar between CF or CF + MS treated pigs and their littermates. These data demonstrate that CF can be used to correct the growth impairment in pigs predicted using plasma AGP as the marker. CF + MS can do the same, but at greater expense.

Simple Summary The rumen is the critical organ mediating nutrient uptake and use in cattle. Health development is essential to ensure animal feed efficiency. In this report, we present an analysis of gene expression dynamic in rumen epithelium during the transition from pre-ruminant to ruminant in cattle fed with hay or concentrated diets at weaning. The global shifts in gene expression and correlated transcription factors activities indicate transcriptional reprogramming during weaning. Transcriptional reprogramming in rumen epithelial tissue reflects critical nutrient-gene interactions occurring during the developmental progression. The results unveiled that nutrient-gene interactions compel transcriptional reprogramming. Our findings also suggest that this transcriptional reprogramming is the molecular basis of the transitional development of pre-ruminant to the ruminant in cattle. Abstract We present an analysis of transcriptomic dynamics in rumen epithelium of 18 Holstein calves during the transition from pre-rumination to rumination in cattle-fed hay or concentrated diets at weaning. Three calves each were euthanized at 14 and 42 d of age to exemplify preweaning, and six calves each were provided diets of either milk replacer and grass hay or calf starter to introduce weaning. The two distinct phases of rumen development and function in cattle are tightly regulated by a series of signaling events and clusters of effectors on critical pathways. The dietary shift from liquid to solid feeds prompted the shifting of gene activity. The number of differentially expressed genes increased significantly after weaning. Bioinformatic analysis revealed gene activity shifts underline the functional transitions in the ruminal epithelium and signify the transcriptomic reprogramming. Gene ontogeny (GO) term enrichment shows extensively activated biological functions of differentially expressed genes in the ruminal epithelium after weaning were predominant metabolic functions. The transcriptomic reprogramming signifies a correlation between gene activity and changes in metabolism and energy production in the rumen epithelium, which occur at weaning when transitioning from glucose use to VFA use by epithelium during the weaning.

Interactions between the bacteria and fungi in the gut microbiome can result in altered nutrition, pathogenicity of infection, and host development, making them a crucial component in host health. Associations between the mycobiome and bacteriome in the piglet gut, in the context of weaning, remain unknown. Weaning is a time of significant stress, dietary changes, microbial alterations, and a predisposition to infection. The loss of animal health and growth makes potential microbial interventions of interest to the swine industry. Recent studies have demonstrated the diversity and development of the microbiome in the gastrointestinal (GI) tract of piglets during weaning, resulting from the dietary and physiological changes. Despite these advances, the role of the mycobiota in piglet health and its contribution to overall microbiome development remains mostly unknown. In this study we investigated the bacteriome and the mycobiome after weaning in the GI tract organs and feces from 35-day old piglets. Following weaning, the α-diversity and amplicon sequence variants (ASVs) counts of the bacteriome increased, proximally to distally, from the stomach to the feces along the GI tract, while the mycobiome α-diversity and ASV counts were highest in the porcine stomach. β-diversity analyses show distinct clusters based on organ type in the bacteriome and mycobiome, but dispersion remained relatively constant in the mycobiome between organ/fecal sites. Bacteroidetes, Firmicutes, and Epsilonbacteraeota were the most abundant bacterial phyla present in the GI tract and feces based on mean taxonomic composition with high variation of composition found in the stomach. In the mycobiome, the dominant phyla were Ascomycota and Basidiomycota, and the stomach mycobiome did not demonstrate the same high level of variation observed in the bacteriome. Potential interactions between genera were found in the lower piglet GI bacteriome and mycobiome with positive correlations found between the fungus, Kazachstania, and several bacterial species, including Lactobacillus. Aspergillus demonstrated negative correlations with the short chain fatty acid-producing bacteria Butyricoccus, Subdoligranulum, and Fusicatenibacter. This study demonstrates the distinct colonization dynamics between fungi and bacteria in the GI tract and feces of piglets directly following weaning and the potential interactions of these microbes in the porcine gut ecosystem.

Consumers' demand of leaner meat products is a challenge. Although betaine and conjugated linoleic acid (CLA) have the potential to decrease porcine adipose tissue, their mode of action is poorly understood. The aim of the study was to determine the lipolytic effect of betaine and CLA in the adipose tissue of Iberian pigs. Adipose tissue explants from five pigs (38 kg BW) were prepared from dorsal subcutaneous adipose tissue samples and cultivated for 2 h (acute experiments) or 72 h (chronic experiments). Treatments included 100 μM linoleic acid (control), 100 μM trans-10, cis-12 CLA, 100 μM linoleic acid + 1 mM betaine and 100 μM trans-10, cis-12 CLA + 1 mM betaine (CLABET). To examine the ability of betaine or CLA to inhibit insulin's suppression of isoproterenol-stimulated lipolysis, test medium was amended with 1 μM isoproterenol ±10 nM insulin. Media glycerol was measured at the end of the incubations. Acute lipolysis (2 h) was increased by CLA and CLABET (85% to 121%; P < 0.05) under basal conditions. When lipolysis was stimulated with isoproterenol (1090%), acute exposure to betaine tended to increase (13%; P = 0.071), while CLA and CLABET increased (14% to 18%; P < 0.05) isoproterenol-stimulated lipolysis compared with control. When insulin was added to isoproterenol-stimulated explants, lipolytic rate was decreased by 50% (P < 0.001). However, supplementation of betaine to the insulin + isoproterenol-containing medium tended to increase (P = 0.07), while CLABET increased (45%; P < 0.05) lipolysis, partly counteracting insulin inhibition. When culture was extended for 72 h, CLA decreased lipolysis under basal conditions (18%; P < 0.05) with no effect of betaine and CLABET (P > 0.10). When lipolysis was stimulated by isoproterenol (125% increase in rate compared with basal), CLA and CLABET decreased glycerol release (27%; P < 0.001) compared with control (isoproterenol alone). When insulin was added to isoproterenol-stimulated explants, isoproterenol stimulation of lipolysis was completely blunted and neither betaine nor CLA altered the inhibitory effect of insulin on lipolysis. Isoproterenol, and especially isoproterenol + insulin, stimulated leptin secretion compared with basal conditions (68% and 464%, respectively; P < 0.001), with no effect of CLA or betaine (P > 0.10). CLA decreased leptin release (25%; P < 0.001) when insulin was present in the media, partially inhibiting insulin stimulation of leptin release. In conclusion, betaine and CLA produced a biphasic response regarding lipolysis so that glycerol release was increased in acute conditions, while CLA decreased glycerol release and betaine had no effect in chronic conditions. Furthermore, CLA and CLABET indirectly increased lipolysis by reducing insulin-mediated inhibition of lipolysis during acute conditions., This research was supported by grants no. AGL 2009-08916 from Ministerio de Educacion y Ciencia and AGL 2016-80231 from Ministerio de Economía y Competitividad, Spain.

Serum α1-acid glycoprotein (AGP) is elevated during late gestation and at birth in the pig and rapidly declines postnatally. In contrast, the pig is born with minimal lipid stores in the adipose tissue, but rapidly accumulates lipid during the first week. The present study examined if AGP can affect adipose tissue metabolism in the neonatal pig. Isolated cell cultures or tissue explants were prepared from dorsal subcutaneous adipose tissue of preweaning piglets. Porcine AGP was used at concentrations of 0, 100, 1000 and 5000 ng/ml medium in 24 h incubations. AGP reduced the messenger RNA (mRNA) abundance of the lipogenic enzymes, malic enzyme (ME), fatty acid synthase and acetyl coA carboxylase by at least 40% (P

Identification of plasma and/or serum markers at birth that will predict animal performance may be useful for identifying animals susceptible to poor growth. Metabolomic analysis of plasma from newborn swine was used to identified potential metabolite differences between 8 pairs of littermates with similar birth weights but whose ADG differed by50 g/d so that, at weaning (21 d), littermates differed in BW by 1.62 kg (P0.01). Plasma analysis failed to identify metabolic pathways impacted by growth, most likely because of the small sample population. Interestingly, despite comparative analysis of 576 metabolites between these slow-growing and normal-growing littermates, the relative abundance of only 36 metabolites differed between the pairs. Most of these metabolites could be eliminated as potential markers because of the difficulty with the extraction and rapid measurement of their plasma/serum concentrations. Histamine differed from most of these potential metabolite markers in that commercial sandwich ELISAs are readily available. Using an ELISA, we verified the metabolomic data, demonstrating that plasma histamine concentrations were 150% higher in slow-growing than normal growing littermates of similar birth weight (P0.05). Subsequently, a separate data set was obtained using swine from a different geographical location and genetic background and also showed that elevated histamine (ng/mL) at birth is associated with increased preweaning growth rate (P = 0.009, r = 0.306, n = 9 litters). Together, the data indicate that perinatal histamine concentrations may serve as a tool to identify potentially slower growing pigs and as a serum biomarker for predicting litter growth rate.

Staphylococcus aureus is an opportunistic pathogen historically considered a minor contributor to porcine illness, but recent work has pointed to the potential of pigs as a source of zoonotic methicillin-resistant S. aureus (MRSA) infections. S. aureus is adept at acquiring antibiotic-resistance genes and has become a pathogen of interest due to its implications in pig and human health. The innate immune response plays a critical role in controlling gram positive bacterial infections (such as S. aureus) in the intestine. To elucidate the acute immune response to gram positive infections, we challenged a porcine intestinal cell line (IPEC-J2) with gram-positive antigens isolated from S. aureus. IPEC-J2 cells were grown in collagen-coated transwell dishes until a transepithelial electrical resistance (TEER) reading of >1 kΩcm(2) was obtained. Cells were then challenged with 100ug/ul peptidoglycan (PGN) or 10ug/ul lipoteichoic acid (LTA) for 3, 8 or 24 hours prior to cell and supernatant collection. Butyrate is a short chain fatty acid in the gut that has known protective effects on barrier function and intestinal health. To test the protective effect of butyrate on gram-positive intestinal infections, IPEC-J2 cells were pre-treated with butyrate for 24 hours prior to PGN or LTA challenge. Pre-treatment with butyrate prior to PGN or LTA challenge resulted in statistically significant increases of IL-1RA. After antigen challenge alone, levels of IL-1RA and IL-8 did not increase significantly over time in the cellular supernatant. IL-1b, IL-4, IL-2, IL-6, IL-10, IL-12, and IL-18 were not significantly altered by antigen challenge +/- butyrate pre-treatment. These data indicate an acute inflammatory response is produced by IPEC-J2 cells upon exposure to antigens from gram-positive bacteria, but pre-treatment with butyrate promotes an anti-inflammatory response, that may reduce intestinal damage, through increased IL-1RA secretion.

Fetuin A (also known as α2-Heremans-Schmid glycoprotein) is a protein primarily expressed by the liver and secreted into the blood. Previous studies have suggested that plasma concentrations of fetuin A are elevated with impaired growth rate in swine. The present study was designed to examine the relationship of porcine fetuin A with growth rate in the pig and to also elucidate the regulation of fetuin A expression by examining the hormonal and cytokine regulation of fetuin A mRNA abundance in hepatocytes prepared from suckling piglets. Quantitative real-time PCR assay was used to quantify the number of fetuin A mRNA molecules/molecule cyclophilin mRNA. Total RNA was isolated from liver of three different groups of pigs to assess changes in mRNA abundance of fetuin A: normal piglets at day 1, day 7 day 21 or 6 months of age (n=6 for each age); runt and control piglets at day 1 of age (n=4); slow growing and normal growing piglets at 21 days of age (n=8). Following birth, fetuin A gene expression increased from day 1 and 7 of age (P

Growth rate affects adipose tissue development and variations in growth rate may potentially impact adipokine expression. Samples of subcutaneous (SQ) and perirenal (PR) adipose tissues and longissimus muscle were collected at day 21 of age from the fastest and slowest growing piglets within seven litters. Reverse transcription and real-time PCR were used to quantify adipokine mRNA abundance. Leptin, adiponectin, tumor necrosis factor α (TNFα ) and lipoprotein lipase (LPL) mRNA abundance were lower in SQ from slow growing piglets (SGP) than in fast growing piglets (FGP, P α gene expression were reduced in PR from SGP in comparison to FGP (P β (IL1β), IL15 and LPL were increased in the longissimus of SGP relative to FGP (P < 0.05). Analysis of mRNA abundance for these adipokines within adipose tissue at day 21 of age demonstrated that the effect of growth rate on adipokine expression varies among different adipokines and the internal and external sites of adipose tissue deposition (PR versus SQ). The increase in longissimus expression of LPL and IL15 suggests that nutrient partitioning for energy use may be greater in the skeletal muscle of the SGP.

Clostridium perfringens is a bacterial pathogen that causes necrotic enteritis in poultry and livestock, and is a source of food poisoning and gas gangrene in humans. As the agriculture industry eliminates the use of antibiotics in animal feed, alternatives to antibiotics will be needed. Bacteriophage endolysins are enzymes used by the virus to burst their bacterial host, releasing bacteriophage particles. This type of enzyme represents a potential replacement for antibiotics controlling C. perfringens. As animal feed is often heat-treated during production of feed pellets, thermostable enzymes would be preferred for use in feed. To create thermostable endolysins that target C. perfringens, thermophile endolysin catalytic domains were fused to cell wall binding domains from different C. perfringens prophage endolysins. Three thermostable catalytic domains were used, two from prophage endolysins from two Geobacillus strains, and a third endolysin from the deep-sea thermophilic bacteriophage Geobacillus virus E2 (GVE2). These domains harbor predicted L-alanine-amidase, glucosaminidase, and L-alanine-amidase activities, respectively and degrade the peptidoglycan of the bacterial cell wall. The cell wall binding domains were from C. perfringens prophage endolysins (Phage LYtic enzymes, Ply): PlyCP18, PlyCP10, PlyCP33, PlyCP41, and PlyCP26F. The resulting fifteen chimeric proteins were more thermostable than the native C. perfringens endolysins, and killed swine and poultry disease-associated strains of C. perfringens.

This study was designed to determine if normal birth weight pigs that grow poorly during the pre-weaning period have altered plasma metabolites that might serve as markers for prediction of growth rate. Eight pairs of average birth weight pigs (1.57 ± 0.05 kg) were identified that diverged in weight by a minimum of 50 g/day until 21 days of age. At 21 days, slow growing (SG) pigs weighed 5.47 ± 0.22 kg while control (C) littermates weighed 6.98 ± 0.28 kg (P < 0.001). Plasma was collected for analysis at days 1 and 21 postpartum for metabolomic analysis by ultrahigh performance liquid chromatography-tandem mass spectroscopy (Metabolon; Durham, NC). Analysis of the plasma from these SG pigs identified 578 metabolites, but only 36 were affected by potential growth rate at day 1 of age (P < 0.05). Plasma analysis at 21 days of age revealed that the concentration of only 28 metabolites were affected by growth rate (P < 0.05). The few plasma metabolite changes did not indicate changes in major metabolic pathways (P > 0.05). Principle component analysis revealed that groups clustered strongly by age, with all day 1 samples strongly segregating along component axis 1 from day 21 samples. Conversely, growth rate had very little effect in further segregating samples, as plasma samples from slow- versus fast-growing pigs forming largely overlapping populations. The results indicate that while age profoundly affects global metabolic profiles, growth rate elicits minimal alterations in global biochemical makeup. Examination of the data for individual plasma metabolites at birth by analysis using 2-way ANOVA with repeated measures identified elevated erythronate (P = 0.009), histamine (P = 0.019), 7-ketolithocholate (P = 0.020), serotonin (P = 0.0257); and s-adenosylhomocysteine (P = 0.001) as potential markers to identify underperforming piglets and their littermates at birth.

This study was designed to determine whether methyl-β-cyclodextrin (MCD) can substitute for albumin in incubation medium for neonatal swine adipose tissue explants, or whether MCD affects metabolism and cytokine expression. Subcutaneous adipose tissue explants (100 ± 10 mg) were prepared from 21-day-old pigs. Explants were incubated in medium 199 supplemented with 25 mM HEPES, 1.0 nM insulin at 37°C. The medium also contained bovine serum albumin (BSA) or MCD at 0%, 0.05%, 0.1%, 0.2% or 0.3%. Tissue explants were treated with these media for 1 h and then switched to the same basal incubation medium containing 0.05% BSA. Explants were removed from basal medium at 2 or 8 h of incubation, and real-time PCR was performed to assess expression of tumor necrosis α (TNF) and interleukin 6 (IL6), acetyl CoA carboxylase (ACAC) and fatty acid synthase (FASN). Alternatively, rates of 14C-glucose oxidation and lipogenesis were monitored ± insulin (100 nM), following MCD treatment. Incubation with BSA had minimal effects on gene expression or adipose tissue metabolism, only producing a doubling in TNF mRNA abundance (P < 0.01). Treatment with MCD increased TNF mRNA abundance by eightfold (P < 0.009), whereas IL6 gene expression increased a 100-fold (P < 0.001) with a suppression in ACAC and FASN expression (P < 0.01). This was paralleled by MCD inhibition of insulin-stimulated glucose oxidation and lipogenesis (P < 0.001). Addition of a TNF antibody to the incubation medium alleviated this inhibition of insulin-stimulated glucose metabolism by ∼30% (P < 0.05).

Background Porcine adipose tissue expresses orosomucoid (ORM1) mRNA, a protein with anti-inflammatory and immunomodulatory properties. Previous research has demonstrated that porcine ORM1 can reduce insulin stimulated glucose metabolism in porcine adipose tissue in vitro. The present study was designed to examine the preweaning ontogeny of ORM1 mRNA abundance in porcine subcutaneous adipose and to determine if ORM1 can regulate mRNA abundance of inflammatory cytokines that contribute to insulin resistance in primary cultures derived from neonatal porcine subcutaneous adipose tissue. Cultures were differentiated in vitro and subsequently the adipocyte containing cultures were incubated for 24 h with 0–5000 ng porcine ORM1/mL medium. Cultures were then harvested, total RNA extracted for use in reverse transcription and the mRNA abundance of cytokine mRNA quantified by real-time PCR. Results ORM1 mRNA abundance within neonatal adipose tissue does not change from d 1 to d 21 of age and is a very small fraction relative to liver mRNA abundance. The ORM1 mRNA level in porcine adipocytes and stromal-vascular cells are similar (P > 0.05). Treatment with ORM1 did not affect TNFα (tumor necrosis factor α) mRNA level (P > 0.05), while interleukin 6 (IL6) mRNA abundance was reduced 32 % at 1,000 ng ORM1/mL (P 0.05). The reduction of macrophage migration inhibitory factor (MIF) mRNA abundance by ORM1 was dose dependent (P

The purpose of this study was to use quantitative magnetic resonance (QMR) to measure changes in the body composition of piglets during growth from birth to 4 kg body weight. Using QMR, 60 pigs were scanned an average of 5 times starting at 2.7±1.3 days of age (1.95 kg) and finally at 13.1±4.3 days (4.14 kg). Regression analysis revealed that the rates of total body growth and fat and lean deposition were linear throughout this period. Subsequently, a second group of 235 pigs (109 males and 126 females) were scanned twice, first at 2.7±1.2 days of age and then at 13.4±3.1 days of age. The mean (±SD) rate of total body growth was 230±57 g/day. The rates of fat and lean deposition were 40±13 g/day and 191±52 g/day, respectively. The rates of both fat and lean deposition were highly correlated (P

The growth and composition of the neonatal pig is of interest because of potential impact on subsequent growth and finally, composition at market weight. The purpose of this study was to compare at weaning the growth and body composition of the largest and smallest pigs from each of 38 litters. At weaning (27±1.7 d) the largest (9.3±1.1 kg) and smallest (6.2±1.5 kg) pigs were selected for body composition measurement by dual energy X-ray absorptiometry (DXA). The body composition of the largest pigs consisted of 38 % more fat, 32 % more lean, and 29 % more bone mineral content (P0.05). A second study consisted of 12 pairs of pigs from 8 litters that were selected on the basis of having the same birth weight, but one pig out gaining the other by at least 50 g/day. At 21 days of age the selected pigs were scanned by DXA. For both groups combined, the correlation (r) between body weight and lean mass was 0.99, between body weight and fat mass it was 0.87, and between body weight at birth and body weight at weaning it was 0.56. The results of these studies revealed that, at weaning, the fastest and slowest growing pigs had similar proportions of fat, lean and bone mineral and, consistent with previous results, the rates of both fat and lean deposition were highly correlated (P

Oxidation of serum proteins can lead to carbonyl formation that alters their function and is often associated with stress-related diseases. As it is recommended that all pigs reared in modern production facilities be given supplemental iron at birth to prevent anemia, and metals can catalyze the carbonylation of proteins, the primary objective of this study was to determine whether standard iron dextran treatment was associated with enhanced serum protein oxidation in newborn piglets. Piglets were treated with 100 mg of iron dextran intramuscularly either on the day of birth, or on the third day after birth. Blood samples were collected from piglets 48 or 96 h after treatment and serum was harvested. For quantification, serum protein carbonyls were converted to hydrazones with dinitrophenyl hydrazine and analyzed spectrophotometrically. To identify and determine relative distribution of carbonylated proteins, serum protein carbonyls were derivatized with biotin hydrazide, separated by two-dimensional polyacrylamide gel electrophoresis, stained with avidin-fluorescein and identified by mass spectrometry. The standard iron dextran treatment was associated with no increase in total oxidized proteins if given either on the first or third day of life. In addition, with a few noted exceptions, the overall distribution and identification of oxidized proteins were similar between control and iron dextran-treated pigs. These results indicate that while iron dextran treatment is associated with a marked increase in circulating iron, it does not appear to specifically induce the oxidation of serum proteins.

ob protein is hypothesized to be a circulating feedback signal in the regulation of energy balance. Obese, overfed rats have high levels of ob mRNA expression and suppressed voluntary food intake, indicating the presence of a potent satiety factor. The objectives of this experiment were to determine whether feeding rats their normal daily intake in three meals, compared with ad libitum feeding, increased ob mRNA expression and to determine the degree of obesity required to stimulate expression of ob mRNA. Rats were fed ad libitum, were tube-fed their normal intake in three meals a day, or were tube-fed twice normal intake, ob mRNA was measured by Northern blot analysis after 0, 2, 7, 14, 21, and 32 d of tube-feeding. After only 2 d ob mRNA was threefold higher in tube-fed animals than in ad libitum controls. By day 21 there was a further increase in ob mRNA expression in overfed rats which were at 130% control weight. These results suggest that a metabolic consequence of meal-feeding increases ob mRNA expression in the absence of increased food intake or weight gain. There is a further increase in ob mRNA expression once significant obesity is established.

article i nfo Oxidation of serum proteins leads to non-reversible carbonyl formation which alters their function and is associated with stress-related disease processes. The primary objective of this study was to quantify and identify oxidized serum proteins in fetal and newborn piglets. Protein carbonyls were converted to hydrazones with dinitrophenyl hydrazine and quantified spectrophotometrically. For identification, serum protein carbonyls were derivatized with biotin hydrazide, separated by 2D PAGE and stained with FITC- avidin. Biotin-labeled proteins were excised from gels and identified by mass spectrometry. At birth, carbonyls were determined to be ∼600 pmole/mg serum protein. Fetuses at 50 and 100 days of gestation had similar levels of protein carbonyls as newborns. Carbonyl levels were also similar for control and runt (b 1k g at birth) piglets between 1 and 21 days of age; however, distribution of many proteins varied by age and was also influenced by birth weight. Major oxidized proteins identified in fetal (f) and newborn (n) pigs included; albumin (f, n), transferrin (f, n), fetuin-A (f, n) alpha fetoprotein (f, n), plasminogen (f, n), fetuin-B (f), alpha-1-antitrypsin (f, n) alpha-1-acid glycoprotein (f) and immunoglobulins (n). While abundance and distribution of oxidized proteins changed over time, these changes appear to primarily reflect relative amounts of those proteins in serum.