Dietary Calcium and Phosphorus Amounts Affect Development and Tissue-Specific Stem Cell Characteristics in Neonatal Pigs.

BACKGROUND: Dietary calcium and phosphorus are required for bone and muscle development. Deficiencies of these macrominerals reduce bone mineral and muscle accretion potentially via alterations of mesenchymal stem cell (MSC) and satellite cell (SC) activities. OBJECTIVES: With increasing interest in the role of early-life events on lifetime health outcomes, we aimed to elucidate the impact of dietary calcium and phosphorus, from deficiency through excess, on MSC and SC characteristics during neonatal development. METHODS: Neonatal pigs [30 females, 1-d-old, 1.46 ± 0.04 kg body weight (BW)] were fed milk replacers for 16 d that were isonitrogenous and isocaloric with a consistent ratio of calcium to phosphorus, but either 25% deficient (calcium: 0.78%; phosphorus: 0.60%; CaPD), adequate (calcium: 1.08%; phosphorus: 0.84%; CaPA), or 25% in excess (calcium: 1.38%; phosphorus: 1.08%; CaPE) of calcium and phosphorus requirements based on sow-milk composition and extrapolation from NRC requirements for older pigs. BW and feed intake were recorded daily. Blood was collected for serum phosphorus, parathyroid hormone (PTH), and fibroblast growth factor 23 (FGF23) determination. Humeri were collected for MSC isolation and radii/ulnae bone were collected for analysis. Longissimus dorsi muscle was collected for SC isolation and analysis. RESULTS: There was 4.6% increase in bone ash percentage in CaPE- versus CaPD-fed pigs (P < 0.05). In vivo proliferation indicated a 41.3% increase in MSCs in CaPA compared with CaPD and a 19% increase in SCs in CaPA compared with both CaPE and CaPD. MSCs from CaPD had 2- to 5-fold greater expression of peroxisome proliferator-activated receptor γ (PPARγ), fatty acid-binding protein 4 (FABP4), and lipoprotein lipase (LPL) but lower osteocalcin (BGLAP) and fibronectin (FN1) expression than CaPA (P < 0.05). SCs from CaPD-fed pigs had 19% lower in vivo proliferation than in CaPA-fed pigs. CONCLUSIONS: These findings demonstrated that feeding a diet marginally deficient in calcium and phosphorus to neonatal pigs had a great impact on bone development, MSC, and SC characteristics. These dietary deficiencies may program future bone health and muscle development by altering MSC and SC activities.

Broad and efficient control of major foodborne pathogenic strains of Escherichia coli by mixtures of plant-produced colicins.

Enterohemorrhagic Escherichia coli (EHEC) is one of the leading causes of bacterial enteric infections worldwide, causing ∼100,000 illnesses, 3,000 hospitalizations, and 90 deaths annually in the United States alone. These illnesses have been linked to consumption of contaminated animal products and vegetables. Currently, other than thermal inactivation, there are no effective methods to eliminate pathogenic bacteria in food. Colicins are nonantibiotic antimicrobial proteins, produced by E. coli strains that kill or inhibit the growth of other E. coli strains. Several colicins are highly effective against key EHEC strains. Here we demonstrate very high levels of colicin expression (up to 3 g/kg of fresh biomass) in tobacco and edible plants (spinach and leafy beets) at costs that will allow commercialization. Among the colicins examined, plant-expressed colicin M had the broadest antimicrobial activity against EHEC and complemented the potency of other colicins. A mixture of colicin M and colicin E7 showed very high activity against all major EHEC strains, as defined by the US Department of Agriculture/Food and Drug Administration. Treatments with low (less than 10 mg colicins per L) concentrations reduced the pathogenic bacterial load in broth culture by 2 to over 6 logs depending on the strain. In experiments using meats spiked with E. coli O157:H7, colicins efficiently reduced the population of the pathogen by at least 2 logs. Plant-produced colicins could be effectively used for the broad control of pathogenic E. coli in both plant- and animal-based food products and, in the United States, colicins could be approved using the generally recognized as safe (GRAS) regulatory approval pathway.

Impact of dietary organic acids and botanicals on intestinal integrity and inflammation in weaned pigs.

BACKGROUND: Organic acids, such as citric and sorbic acid, and pure plant-derived constituents, like monoterpens and aldehydes, have a long history of use in pig feeding as alternatives to antibiotic growth promoters. However, their effects on the intestinal barrier function and inflammation have never been investigated. Therefore, aim of this study was to assess the impact of a microencapsulated mixture of citric acid and sorbic acid (OA) and pure botanicals, namely thymol and vanillin, (PB) on the intestinal integrity and functionality of weaned pigs and in vitro on Caco-2 cells. In the first study 20 piglets were divided in 2 groups and received either a basal diet or the basal diet supplemented with OA + PB (5 g/kg) for 2 weeks post-weaning at the end of which ileum and jejunum samples were collected for Ussing chambers analysis of trans-epithelial electrical resistance (TER), intermittent short-circuit current (I SC), and dextran flux. Scrapings of ileum mucosa were also collected for cytokine analysis (n = 6). In the second study we measured the effect of these compounds directly on TER and permeability of Caco-2 monolayers treated with either 0.2 or 1 g/l of OA + PB. RESULTS: Pigs fed with OA + PB tended to have reduced I SC in the ileum (P = 0.07) and the ileal gene expression of IL-12, TGF-ß, and IL-6 was down regulated. In the in vitro study on Caco-2 cells, TER was increased by the supplementation of 0.2 g/l at 4, 6, and 14 days of the experiment, whereas 1 g/l increased TER at 10 and 12 days of treatment (P < 0.05). Dextran flux was not significantly affected though a decrease was observed at 7 and 14 days (P = 0.10 and P = 0.09, respectively). CONCLUSIONS: Overall, considering the results from both experiments, OA + PB improved the maturation of the intestinal mucosa by modulating the local and systemic inflammatory pressure ultimately resulting in a less permeable intestine, and eventually improving the growth of piglets prematurely weaned.

Dietary calcium deficiency and excess both impact bone development and mesenchymal stem cell lineage priming in neonatal piglets.

BACKGROUND: Optimizing calcium nutrition to maximize bone accretion during growth to prevent fragility fractures later in life has spurred greater interest in calcium nutrition in neonates. OBJECTIVE: The aim of this study was to determine the effect of dietary calcium, from deficiency through excess, on bone growth, and the in vivo and in vitro behavior of mesenchymal stem cells (MSCs) in neonatal pigs. METHODS: Twenty-four male and female piglets (24 ± 6 h old) were fed either a calcium-deficient [Ca-D; 0.6% Ca on a dry matter (DM) basis], a calcium-adequate diet (Ca-A; 0.9% Ca on a DM basis), or a calcium-excessive diet (Ca-E; 1.3% Ca on a DM basis) for 14 d to assess the impact of dietary calcium on calcium homeostasis and on the behavior of MSCs. RESULTS: Growth rate was not affected by the Ca-E diet, although bone ash content was 16% higher (P < 0.05) and urinary calcium excretion was 5-fold higher, when normalized to creatinine, compared with the Ca-A group at trial completion. Serum parathyroid hormone (PTH) concentrations were elevated (P < 0.05) in Ca-D piglets in comparison with other groups at both 7 and 14 d. In vivo proliferation of MSCs was 30% higher (P < 0.05) in Ca-E piglets than the other groups. MSCs from both Ca-D- and Ca-E-fed piglets had greater adipogenic potential based on increased gene expression (P < 0.05) of peroxisome proliferator-activated receptor γ (Pparg) and adipocyte fatty acid-binding protein (Ap2) than MSCs from Ca-A piglets. Interestingly, only MSCs from Ca-E-fed piglets had greater (P < 0.05) gene expression of lipoprotein lipase (Lpl) during adipocytic differentiation than those from Ca-A piglets. To assess alterations in lineage allocation and priming, the most and least osteogenic (O+ and O-, respectively) and adipogenic (A+ and A-, respectively) colonies from each MSC isolation were selected on the basis of functional staining. The O+ colonies from Ca-D piglets expressed lower (P < 0.05) levels of osteocalcin (OC) mRNA than did those from other groups, whereas the O- colonies from Ca-E piglets expressed higher (P < 0.05) levels of mRNA of Pparg, Ap2, and Lpl than did those from other groups. CONCLUSIONS: Neonatal calcium deficiency appears to reduce the osteogenic priming of MSCs while enlarging a subpopulation of potentially adipogenic cells, and excess dietary calcium appears to allow greater multipotency of MSCs. These programming alterations of MSCs could have long-term consequences for bone health.

Calcium trafficking and gastrointestinal physiology following an acute lipopolysaccharide challenge in pigs.

The influence of systemic immune activation on whole-body calcium (Ca) trafficking and gastrointestinal tract (GIT) physiology is not clear. Thus, the study objectives were to characterize the effects of lipopolysaccharide (LPS) on Ca pools and GIT dynamics to increase understanding of immune-induced hypocalcemia, ileus, and stomach hemorrhaging. Twelve crossbred pigs [44 ±â€…3 kg body weight (BW)] were randomly assigned to 1 of 2 intramuscular treatments: (1) control (CON; 2 mL saline; n = 6) or (2) LPS (40 µg LPS/kg BW; n = 6). Pigs were housed in metabolism stalls to collect total urine and feces for 6 h after treatment administration, at which point they were euthanized, and various tissues, organs, fluids, and digesta were weighed, and analyzed for Ca content. Data were analyzed with the MIXED procedure in SAS 9.4. Rectal temperature and respiration rate increased in LPS relative to CON pigs (1.4 °C and 32%, respectively; P ≤ 0.05). Inflammatory biomarkers such as circulating alkaline phosphatase, aspartate aminotransferase, and total bilirubin increased in LPS compared with CON pigs whereas albumin decreased (P ≤ 0.02). Plasma glucose and urea nitrogen decreased and increased, respectively, after LPS (43% and 80%, respectively; P < 0.01). Pigs administered LPS had reduced circulating ionized calcium (iCa) compared to CON (15%; P < 0.01). Considering estimations of total blood volume, LPS caused an iCa deficit of 23 mg relative to CON (P < 0.01). Adipose tissue and urine from LPS pigs had reduced Ca compared to CON (39% and 77%, respectively; P ≤ 0.05). There did not appear to be increased Ca efflux into GIT contents and no detectable increases in other organ or tissue Ca concentrations were identified. Thus, while LPS caused hypocalcemia, we were unable to determine where circulating Ca was trafficked. LPS administration markedly altered GIT dynamics including stomach hemorrhaging, diarrhea (increased fecal output and moisture), and reduced small intestine and fecal pH (P ≤ 0.06). Taken together, changes in GIT physiology suggested dyshomeostasis and alimentary pathology. Future research is required to fully elucidate the etiology of immune activation-induced hypocalcemia and GIT pathophysiology.

Lipopolysaccharide (LPS) activates the immune system and this is accompanied with hypocalcemia and altered gastrointestinal tract (GIT) physiology. The study objectives were to characterize whole-body calcium (Ca) trafficking and evaluate GIT dynamics during LPS-induced immune activation. Ca concentrations were analyzed after intramuscular LPS injection. Administering LPS caused marked alterations in metabolic and inflammatory biomarkers and GIT dynamics, characterized by increased lower GIT motility and stomach hemorrhaging. Circulating Ca and adipose tissue and urine Ca output were decreased after LPS. Ca concentrations in other tissues and GIT contents were not detectably different. Thus, we were unable to account for about 110 mg Ca following LPS. Where and how circulating Ca is partitioned during immune activation remains unclear.

Intestinal Damages by F18+Escherichia coli and Its Amelioration with an Antibacterial Bacitracin Fed to Nursery Pigs.

This study investigated intestinal oxidative damage caused by F18+Escherichia coli and its amelioration with antibacterial bacitracin fed to nursery pigs. Thirty-six weaned pigs (6.31 ± 0.08 kg BW) were allotted in a randomized complete block design. Treatments were: NC, not challenged/not treated; PC, challenged (F18+E. coli at 5.2 × 109 CFU)/not treated; AGP challenged (F18+E. coli at 5.2 × 109 CFU)/treated with bacitracin (30 g/t). Overall, PC reduced (p < 0.05) average daily gain (ADG), gain to feed ratio (G:F), villus height, and villus height to crypt depth ratio (VH:CD), whereas AGP increased (p < 0.05) ADG, and G:F. PC increased (p < 0.05) fecal score, F18+E. coli in feces, and protein carbonyl in jejunal mucosa. AGP reduced (p < 0.05) fecal score and F18+E. coli in jejunal mucosa. PC reduced (p < 0.05) Prevotella stercorea populations in jejunal mucosa, whereas AGP increased (p < 0.05) Phascolarctobacterium succinatutens and reduced (p < 0.05) Mitsuokella jalaludinii populations in feces. Collectively, F18+E. coli challenge increased fecal score and disrupted the microbiota composition, harming intestinal health by increasing oxidative stress, and damaging the intestinal epithelium, ultimately impairing growth performance. Dietary bacitracin reduced reduced F18+E. coli populations and the oxidative damages they cause, thereby improving intestinal health and the growth performance of nursery pigs.

Dietary calcium restriction affects mesenchymal stem cell activity and bone development in neonatal pigs.

The effects of dietary calcium (Ca) deficiency on skeletal integrity are well characterized in growing and mature mammals; however, less is known about Ca nutrition during the neonatal period. In this study, we examined the effects of neonatal Ca nutrition on bone integrity, endocrine hormones, and mesenchymal stem cell (MSC) activity. Neonatal pigs (24 ± 6 h of age) received either a Ca-adequate (1.2 g/100 g) or an ~40% Ca-deficient diet for 18 d. Ca deficiency reduced (P < 0.05) bone flexural strength and bone mineral density without major differences in plasma indicators of Ca status. There were no meaningful differences in plasma Ca, phosphate (PO(4)), parathyroid hormone, or 1,25-dihydroxycholecalciferol due to Ca nutrition throughout the study. Calcium deficiency also reduced (P < 0.05) the in vivo proliferation of MSC by ~50%. In vitro studies utilizing homologous sera demonstrated that MSC activity was affected (P < 0.05) by both the Ca status of the pig and the sera as well as by their interaction. The results indicate that neonatal Ca nutrition is crucial for bone integrity and suggest that early-life Ca restriction may have long-term effects on bone integrity via programming of MSC.

Evaluation of phage treatment as a strategy to reduce Salmonella populations in growing swine.

Salmonella is a foodborne pathogenic bacterium that causes human illnesses and morbidity and mortality in swine. Bacteriophages are viruses that prey on bacteria and are naturally found in many microbial environments, including the gut of food animals, and have been suggested as a potential intervention strategy to reduce Salmonella levels in the live animal. The present study was designed to determine if anti-Salmonella phages isolated from the feces of commercial finishing swine could reduce gastrointestinal populations of the foodborne pathogen Salmonella Typhimurium in artificially inoculated swine. Weaned pigs (n = 48) were randomly assigned to two treatment groups (control or phage-treated). Each pig was inoculated with Salmonella Typhimurium (2 × 10(10) colony forming units/pig) via oral gavage at 0 h and fecal samples were collected every 24 h. Swine were inoculated with a phage cocktail via oral gavage (3 × 10(9) plaque forming units) at 24 and 48 h. Pigs were humanely killed at 96 h, and cecal and rectal intestinal contents were collected for quantitative and qualitative analysis. Fecal Salmonella populations in phage-treated pigs were lower (p < 0.09) than controls after 48 h. Phage treatment reduced intestinal populations of inoculated Salmonella Typhimurium in pigs compared to controls at necropsy. Cecal populations were reduced (p = 0.07) by phage treatment >1.4 log(10) colony forming units/g digesta, and rectal populations were numerically reduced. The number of pigs that contained inoculated Salmonella Typhimurium was reduced by phage treatment, but a significant (p < 0.05) reduction was only observed in the rectum. We conclude that phages can be a viable tool to reduce Salmonella in swine. Further research needs to be performed to determine the most efficacious dosing regimens and the most effective combinations of phages targeting the diverse Salmonella population found in swine before they can enter the food supply.

Tributyrin, a Butyrate Pro-Drug, Primes Satellite Cells for Differentiation by Altering the Epigenetic Landscape.

Satellite cells (SC) are a population of muscle resident stem cells that are responsible for postnatal muscle growth and repair. With investigation into the genomic regulation of SC fate, the role of the epigenome in governing SC myogenesis is becoming clearer. Histone deacetylase (HDAC) inhibitors have been demonstrated to be effective at enhancing the myogenic program of SC, but their role in altering the epigenetic landscape of SC remains undetermined. Our objective was to determine how an HDAC inhibitor, butyrate, promotes myogenic differentiation. SC from tributyrin treated neonatal piglets showed a decrease relative to SC from control animals in the expression of enhance of zeste homologue-2 (EZH2), a chromatin modifier, ex vivo. Chromatin Immunoprecipitation-Sequencing (ChIP-Seq) analysis of SC isolated from tributyrin treated pigs showed a global reduction of the tri-methylation of lysine 27 of histone H3 (H3K27me3) repressive chromatin mark. To determine if reductions in EZH2 was the primary mechanism through which butyrate affects SC behavior, SC were transfected with siRNA targeting EZH2, treated with 0.5 mM butyrate, or both. Treatment with butyrate reduced paired-box-7 (Pax7) and myogenic differentiation-1 (MyoD) gene expression, while siRNA caused reductions in EZH2 had no effect on their expression. EZH2 depletion did result in an increase in differentiating SC, but not in myotube hypertrophy. These results indicate that while EZH2 reduction may force myogenic differentiation, butyrate may operate through a parallel mechanism to enhance the myogenic program.

Dietary phosphate restriction decreases stem cell proliferation and subsequent growth potential in neonatal pigs.

Although mesenchymal stem cells (MSC) and satellite cells are essential for postnatal muscle and bone development and phosphate (PO(4)) restriction reduces both muscle and skeletal tissue growth, no research to our knowledge has investigated the possible mechanism by which this mineral may affect early cell programming. Twenty piglets obtained at 1 d of age (1.8 +/- 0.3 kg) received either a PO(4)-adequate diet or a 25% less PO(4)-available diet over a 15-d trial. Feed intake and body weight were recorded daily and blood samples collected every 5 d. After 15 d, pigs were given an intraperitoneal injection of bromodeoxyuridine 4 h prior to tissue collection. As expected, PO(4) deficiency resulted in reduced growth (P < 0.05), feed conversion efficiency (P < 0.05), and bone mineral content (P < 0.05), as well as lower plasma concentrations of both PO(4) (P < 0.01) and parathyroid hormone (P < 0.05). In addition to these classical indicators of PO(4) deficiency, there was also reduced proliferation of both MSC (P < 0.01) and satellite cells (P < 0.05) in vivo. The expression of osteocalcin mRNA in bone marrow was also 2-fold greater (P < 0.01) within the PO(4)-adequate treatment group. These data indicate that in addition to reductions in muscle and bone growth, dietary PO(4) affects proliferation of tissue-specific stem cells in vivo. Nutritional programming of tissue-specific stem cells by dietary PO(4) may have profound implications for life-long growth potential.

Occurrence of Salmonella-specific bacteriophages in swine feces collected from commercial farms.

Salmonella is one of the leading causes of human foodborne illness and is associated with swine production. Bacteriophages are naturally occurring viruses that prey on bacteria and have been suggested as a potential intervention strategy to reduce Salmonella levels in food animals on the farm and in the lairage period. If phages are to be used to improve food safety, then we must understand the incidence and natural ecology of both phages and their hosts in the intestinal environment. This study investigates the incidence of phages that are active against Salmonella spp. in the feces of commercial finishing swine. Fecal samples (n = 60) were collected from each of 10 commercial swine finishing operations. Samples were collected from 10 randomly selected pens throughout each operation; a total of 600 fecal samples were collected. Salmonella spp. were found in 7.3% (44/600) of the fecal samples. Bacteriophages were isolated from fecal samples through two parallel methods: (1) initial enrichment in Salmonella Typhimurium; (2) initial enrichment in Escherichia coli B (an indicator strain), followed by direct spot testing against Salmonella Typhimurium. Bacteriophages active against Salmonella Typhimurium were isolated from 1% (6/600) of the individual fecal samples when initially enriched in Salmonella Typhimurium, but E. coli B-killing phages were isolated from 48.3% (290/600) of the fecal samples and only two of these phages infected Salmonella Typhimurium on secondary plating. Collectively, our results indicate that bacteriophages are widespread in commercial swine, but those capable of killing Salmonella Typhimurium may be present at relatively low population levels. These results indicate that phages (predator) populations may vary along with Salmonella (prey) populations; and that phages could potentially be used as a food safety pathogen reduction strategy in swine.

Characterizing the amount and variability of intramuscular fat deposition throughout pork loins using barrows and gilts from two sire lines.

The objective was to determine the amount and variability of intramuscular fat (IMF) in a pork loin attributable to anatomical chop location, sex, and sire line. Pigs were sired by commercially available terminal Duroc boars selected for meat quality (MQ; n = 96) or lean growth (LG; n = 96) and equally split between barrows and gilts. After slaughter and fabrication, bone-in chops were removed from four locations of each left-side loin (A = 6th rib, B = 10th rib, C = last rib, and D = 4th lumbar vertebrae). An adjacent pair of chops from each location was collected and evaluated for visual color and marbling, subjective firmness, moisture and extractable lipid (IMF) (anterior chop), and Warner-Bratzler shear force (posterior chop). Data were analyzed using the MIXED procedure of SAS as a split-plot design. Homogeneity of variances was tested on raw data using Levene's test of the GLM procedure and found to be heterogeneous. Thus, a two-variance model was fit using the REPEATED statement of the MIXED procedure, grouped by pig. The mivque(0) option of the VARCOMP procedure was used to calculate the proportion of variability that each factor contributed to the total variance. Barrows (3.64%) produced chops with greater (P < 0.01) IMF content than gilts (3.20%), and barrows (2.14) had greater (P < 0.01) IMF variability than gilts (1.23). Chops from MQ pigs (4.02%) exhibited greater (P < 0.01) IMF content than LG (2.82%), and MQ (1.76) had greater IMF variability (P < 0.01) than LG pigs (0.97). Chops from locations A (3.80%) and D (3.77%) had greater IMF than B (3.34%; P < 0.01), and A, B, and D had greater IMF than C (2.77%; P < 0.01). Variances of IMF also differed (A = 1.44, B = 1.59, C = 1.05, and D = 2.18; P = 0.01) across chop locations. Of the variability in IMF, 33.0% was attributed to sire line, 10.16% to chop location, and 4.01% to sex, with 52.83% not accounted for by these three factors. Location A chops were the most (P < 0.01) tender (2.57 kg) and C chops the least (P < 0.01) tender (2.93 kg), while B and D chops were intermediate and not different from each other. No differences in variability (P = 0.40) of tenderness were observed among chop locations (A = 0.31, kg B = 0.24 kg, C = 0.24 kg, and D = 0.23 kg). These results demonstrated that variability in tenderness values did not reflect the variability of IMF. In conclusion, chop location, sex, and sire line all contribute to the amount and variability of pork loin marbling.

Butyric acid induces spontaneous adipocytic differentiation of porcine bone marrow-derived mesenchymal stem cells.

Butyric acid (BA) affects the differentiation of mesenchymal stem cells (MSC) through the activation of different transcriptional pathways. The aim of this study was to determine the effects of BA on proliferation and spontaneous differentiation of porcine bone marrow-derived MSC. Second passage MSC (n = 6) were cultured in either a basal medium (BM, DMEM + 10% FBS), or BM + 2.5 mmol/L BA (BA-2.5) or BM + 5 mmol/L BA (BA-5). Cell proliferation was significantly decreased by both BA-2.5 and BA-5 after 48 h and 72 h (- 55% and - 63%, respectively). To assess the impact of BA on spontaneous differentiation, MSC were cultured for 27 d, with complete media changes every 3 d. At day 27, cells were stained for osteocytic, chondrocytic, and adipocytic differentiation. No terminal differentiation was detected in control MSC, while accumulated small drops of lipids were stained by Oil-Red-O in BA-treated cells. The phenotypic changes were associated with changes in gene expression, determined by qPCR. Treatment with BA modulated the expression of adipocytic differentiation markers: peroxisome proliferator-activated receptor γ and CCAAT/enhancer binding protein α were significantly increased by both BA-2.5 and BA-5 throughout the study, while lipoprotein lipase and fatty acid-binding protein 4 were increased by BA-5 at day 3, and decreased by both BA-5 and BA-2.5 later throughout the study. Osteocalcin and aggrecan mRNA was reduced throughout the experiment by both doses of BA (P < 0.05). In conclusion, our data support that BA promotes the spontaneous differentiation of porcine bone marrow-derived MSC toward an adipocytic lineage in the absence of inducing cocktail media.

Physical activity prevents augmented body fat accretion in moderately iron-deficient rats.

Recent studies describe an association between poor iron status and obesity in humans, although the mechanism explaining this relationship is unclear. The present study aimed to determine the effect of moderate iron deficiency and physical activity (PA) on body composition in an animal model. Male Sprague-Dawley rats consumed iron-adequate (IA; 40 mg/kg) or moderately iron-deficient (ID; 9 mg/kg) diets ad libitum for 12 wk. Rats were assigned to 4 treatment groups (n = 10 per group): IA, sedentary (IAS); IA, PA (IAPA); ID, sedentary (IDS); or ID, PA (IDPA). Activity involved running on motorized running wheels at 4 m/min for 1 h/d for 5 d/wk. After 12 wk, ID rats were not anemic, but body iron stores were reduced as indicated by diminished (P < 0.05) femur iron compared with IA rats. Treatment group did not affect body weight or feed consumption. However, fat mass was greater (P < 0.05) in IDS rats (38.6 +/- 6.7%) than IAS (31.8 +/- 2.9%), IAPA (31.8 +/- 2.0%), and IDPA (32.8 +/- 4.5%) rats. Furthermore, lean body mass was diminished in IDS rats (58.7 +/- 6.8%) compared with IAS (65.6 +/- 3.0%), IAPA (65.6 +/- 2.1%), and IDPA (64.7 +/- 4.5%) rats. Thus, moderate iron deficiency may cause increased body fat accretion in rats and PA attenuates that effect.

Application of colicin E1 as a prefabrication intervention strategy.

Colicin E1 (ColE1) is a bacteriocin produced by and effective against Escherichia coli and related species. The current study examined ColE1 as a potential intervention strategy for controlling E. coli O157:H7 contamination on beef carcasses. Untrimmed beef round roasts were cut into sample sizes of 5.08 by 2.52 by 5.08 cm, with an adipose layer covering an entire surface of lean beef. Samples were placed on sterile metal hooks and inoculated with E. coli O157:H7 at a level of 5 log CFU/ml in sterile tryptic soy broth. After inoculum attachment, ColE1 in doses of 0, 100 microg, 500 microg, and 1 mg/ml of 10 mM Tris, pH 7.6, was sprayed on the samples for a period of 10 min. Samples were evaluated at 0 and 30 min, 1, 2, 3, 4, and 5 days post-spraying at 10 degrees C for E. coli O157:H7 inhibition. Treating samples with 500 microg and 1 mg of ColE1 effectively inhibited E. coli O157:H7 growth. When these doses were applied to samples inoculated with E. coli WS 3331, E. coli contamination was reduced by 4 and 7 log CFU/cm2, respectively, compared with the untreated control samples. In strain WS 3331, treatment with 1 mg ColE1 significantly inhibited growth of E. coli O157:H7 compared with the untreated control during the entire study. ColE1 provided powerful reduction of E. coli O157:H7 as a beef carcass spray intervention.

Dietary tributyrin, an HDAC inhibitor, promotes muscle growth through enhanced terminal differentiation of satellite cells.

Muscle growth and repair rely on two main mechanisms - myonuclear accretion and subsequent protein accumulation. Altering the ability of muscle resident stem cells (satellite cells) to progress through their myogenic lineage can have a profound effect on lifetime muscle growth and repair. The use of the histone deacetylase (HDAC) inhibitor, butyrate, has had positive outcomes on the in vitro promotion of satellite cell myogenesis. In animal models, the use of butyrate has had promising results in treating myopathic conditions as well as improving growth efficiency, but the impact of dietary butyrate on satellite cells and muscle growth has not been elucidated. We investigated the impact of tributyrin, a butyrate prodrug, on satellite cell activity and muscle growth in a piglet model. Satellite cells from tributyrin-treated piglets had altered myogenic potential, and piglets receiving tributyrin had a ~40% increase in DNA:protein ratio after 21 days, indicating the potential for enhanced muscle growth. To assess muscle growth potential, piglets were supplemented tributyrin (0.5%) during either the neonatal phase (d1-d21) and/or the nursery phase (d21-d58) in a 2 × 2 factorial design. Piglets who received tributyrin during the neonatal phase had improved growth performance at the end of the study and had a ~10% larger loin eye area and muscle fiber cross-sectional area. Tributyrin treatment in the nursery phase alone did not have a significant effect on muscle growth or feed efficiency. These findings suggest that tributyrin is a potent promoter of muscle growth via altered satellite cell myogenesis.

Correlation comparisons among early postmortem loin quality and aged loin and pork chop quality characteristics between finishing pigs from either Duroc or Pietrain sires.

Today, the United States exports 2.2 million tons of pork and pork products annually, representing just over 26% of U.S. pork production. In order to meet specific demands of a growing export market, pork quality and carcass characteristics are now integrated into breeding objectives. Color and marbling are 2 loin quality traits that influence consumer acceptability of pork and while correlations between early and aged ventral quality have been established, it is unclear if those correlations differ between production objectives (meat quality vs. lean growth). Therefore, the objective of this experiment was to compare correlations among early postmortem ventral loin quality characteristics and aged ventral loin and chop quality characteristics between pigs sired by either Pietrain (lean growth) or Duroc (meat quality) boars. Early postmortem (~1 d) quality traits included: instrumental and visual color, marbling and firmness, and loin pH on the ventral surface of the loin. Loins were aged until 14 d postmortem in vacuum packages. Aged quality traits included traits evaluated early as well as Warner-Bratzler shear force (WBSF) and cook loss. Correlations were compared between Pietrain and Duroc-sired pigs using a Fisher's z-test. Early instrumental lightness (L*) was moderately correlated with aged ventral L* (Pietrain r = 0.47; Duroc r = 0.65) and aged ventral visual color (Pietrain r = 0.42; Duroc r = 0.58). Early ventral visual color was moderately correlated with aged chop L* (Pietrain r = 0.46; Duroc r = 0.60) and aged chop visual color (Pietrain r = 0.45; Duroc r = 0.57). Early visual marbling was strongly correlated (Pietrain r = 0.68; Duroc r = 0.84) with aged chop visual marbling. Within the Duroc-sired pigs, early L* was moderately correlated with aged chop L* (r = 0.64) but only weakly correlated (r = 0.35) within the Pietrain-sired pigs and those correlations differed at P ≤0.02. Within the Duroc-sired pigs, early ventral visual color was moderately correlated with aged pH (r = 0.44) and aged ventral L* (r = 0.57) but only weakly correlated (r ≤ 0.29) within the Pietrain-sired pigs and those correlations differed at P ≤0.03. No early postmortem quality traits were correlated (|r| ≤ 0.34) with WBSF or cook loss for either sire line. In summary, correlations between early and aged postmortem quality traits rarely differed between Duroc- and Pietrain-sired pigs. It is not necessary to account for sire line when relating early and aged quality characteristics.

Moderate zinc deficiency negatively affects biomechanical properties of rat tibiae independently of body composition.

To guide development of novel nutritional strategies aimed at reducing the incidence of stress fractures, we observed the effects of manipulating dietary zinc (Zn) content on bone integrity in Sprague-Dawley rats fed either a severely Zn-deficient (ZnD; 1 ppm), a moderately Zn-deficient (MZnD; 5 ppm) or a Zn-adequate (ZnAD; 30 ppm) diet for 6 weeks. At the completion of the diet period, body composition, bone mineral content (BMC), bone area (BA) and bone mineral density (BMD) were determined in vivo by using dual-energy X-ray absorptiometry. Following euthanasia, long bones were collected for determination of Zn content and biomechanical strength testing. Despite significant positive correlations between dietary Zn and both body weight (BW) and bone Zn content for the entire cohort (r = .77 and r = .83, respectively), rats fed MZnD or ZnAD diets did not differ in feed intakes, body composition, BMC, BA, BMD or BW. Tibial bones, but not femur bones, appear to be more responsive to dietary Zn manipulation, as all bone biomechanical strength indices in the ZnAD-fed rats were significantly greater than in rats fed the ZnD diets. Rats fed either MZnD or ZnAD diets had stronger tibiae (129% increase in maximum load and stress at maximum load, P<.01) compared with those fed ZnD diets. The load at breakage for the tibial bones of rats fed MZnD diets was not different from the ZnD rats, but lower (P<.05) than that of the ZnAD rats. These results suggest that since feed intakes, body composition, BMC, BA, BMD and BW were not significantly different between the MZnD- and ZnAD-fed animals, the reduced bone integrity observed in the MZnD-fed rats resulted from dietary Zn inadequacy, and not as a result of the reduced growth that is typically associated with Zn deficiency.

Inhibitory activity of colicin E1 against Listeria monocytogenes.

Colicins are gram-negative bacteriocins produced by and effective against Escherichia coli and related species. Colicin E1 (ColE1) is composed of three functional domains, which collectively have a pore-forming effect on targeted bacteria. ColE1 binding and translocation domains are highly specific in contrast to the pore-forming domain, implying that ColE1 could be broadly effective. In this study, the activity of ColE1 against Listeria monocytogenes was evaluated in broth and on surfaces of ready-to-eat products. Individual strains of L. monocytogenes were examined in broth containing ColE1 at 0, 0.1, 1, or 10 microg/ml. Although strain differences in sensitivity to ColE1 existed, growth was significantly reduced in all strains at doses as low as 0.1 microg/ml. Sterilized ham slices were submerged in a five-strain L. monocytogenes cocktail (either 7 or 4 log CFU/ ml) and placed in vacuum packages containing 0, 1, 5, 10, 25, or 50 microg of ColE1. Ham slices were then stored at 4 or 10 degrees C, and samples were removed and examined for L. monocytogenes after 1, 3, 7, and 14 days. Reduction of L. monocytogenes by ColE1 was dependent on initial inoculum concentration and storage temperature. For slices stored at 4 degrees C, treatment with 25 microg reduced Listeria growth below detection limits for the slices inoculated with 4 log CFU/ml for the entire 14 days, whereas for the 7-log CFU/ml slices, growth was detected at 7 days postinoculation. For slices stored at 10 degrees C, 10 microg/ml ColE1 significantly inhibited growth of L. monocytogenes for up to 3 days for both inoculation groups. These data indicate that ColE1 is highly effective against Listeria.

Genetic background influences metabolic response to dietary phosphorus restriction.

Dietary phosphorus (P) is essential to bone growth and turnover; however, little research has focused on the genetic mechanisms controlling P utilization. Understanding the interactions between genetics and dietary P that optimize bone integrity could provide novel interventions for osteoporosis. Thirty-six pigs from two sire lines known to differ in bone structure [heavier boned (HB) and lighter boned (LB)] were assigned to one of the three diets (P adequate, P repletion or P deficient). After 14 days, bone marrow and intact radial bones were collected. Differences between these lines in growth rate, bone integrity and gene expression within bone marrow were observed. In HB, but not LB, pigs, the P-deficient diet decreased weight gain (P<.01). For both lines, P deficiency caused a reduction in radial bone strength (P<.01), but HB P-deficient animals had greater (P<.10) bone integrity than P-deficient LB pigs. In HB, but not LB, pigs, dietary treatment affected the expression of CALCR (calcitonin receptor) (P<.05), VDR (vitamin D receptor) (P<.04) and IGFBP3 (insulin-like growth factor binding protein 3) (P<.06). There was also a trend of increased IL6 (interleukin-6), TFIIB (transcription initiation factor IIB) and SOX9 (sex determining region Y-box 9) expression with P deficiency in HB, but not LB, pigs. Both genetic backgrounds responded similarly to P deficiency with an increase in the expression of OXTR (oxytocin receptor) and IGF1 (insulin-like growth factor 1). Differences in growth rate, bone integrity and gene expression within the bone marrow suggest a difference in the homeorhetic control of P utilization between these genetic lines. Understanding these differences could lead to novel treatments for osteoporosis and aid in the development of tests for identifying those at risk for this disease.