We evaluated the preferences of nursery pigs for diets containing increasing distillers dried grains with solubles (DDGS), varying in color, or high-protein distillers dried grains (HP-DDG) and the effects of flavor supplementation on pig preference and growth performance. In Exp. 1 through 5, diet preference was determined in weanling pigs adjusted to a commercial diet for at least 10 d, and then housed individually for a 2-d double-choice preference test. In Exp. 1, a total of 60 pigs (11.6 ± 0.3 kg of BW) were given a choice between a reference diet (0% DDGS) and test diets containing 0, 10, 20, or 30% DDGS. In Exp. 2, a total of 80 pigs (10.8 ± 0.1 kg of BW) were given a choice between a reference diet (0% HP-DDG) and diets containing 0, 10, 20, or 30% HP-DDG. In Exp. 3, a total of 80 pigs (10.3 ± 0.2 kg of BW) were given a choice between a reference diet (0% DDGS) and a diet containing 0%, 30% light, or 30% dark DDGS. In Exp. 4, a total of 80 pigs (11.2 ± 0.2 kg of BW) were given a choice between a reference diet without DDGS and a diet containing either 0% DDGS, 10 or 20% light DDGS, or 10 or 20% dark DDGS. In Exp. 5, a total of 108 pigs (9.0 ± 0.2 kg of BW) were given a choice between a reference diet (0% DDGS and no flavor) and a diet without or with flavor and containing 0, 10, or 20% DDGS. In Exp. 1 and 2, DDGS and HP-DDG, respectively, linearly decreased (P < 0.01) pig preference. In Exp. 3, dark DDGS were preferred (P < 0.05) compared with light DDGS. In Exp. 4, preferences were linearly reduced (P < 0.01) with DDGS inclusion, and dark DDGS tended (P = 0.06) to be preferred compared with light DDGS. In Exp. 5, DDGS reduced preference (P < 0.01) and flavor reduced preference (P < 0.01) regardless of DDGS level. In Exp. 6, a total of 192 pigs (6.7 ± 0.1 kg of BW) were fed starter 1 diets without or with flavor for 1 wk. Subsequently, pigs were fed starter 2 and 3 diets (2 wk each) containing 0, 10, or 20% DDGS while continuing to receive their respective flavor treatment. Flavor addition during the starter 1 phase increased ADFI (P = 0.02), and DDGS inclusion tended to decrease ADG (P = 0.06) and decreased ADFI (P = 0.03) during the starter 2 phase. Volatile components in DDGS and HP-DDG varied greatly depending on the source. Nursery pigs preferred a diet without DDGS or HP-DDG, and this appeared to be unrelated to color differences between sources. Knowledge of volatile compounds that enhance or suppress the palatability of feed may lead to further development of feed additives for masking relatively unpalatable, albeit cost-effective, ingredients.
In an effort to develop a sustainable platform for manufacturing protein-based vaccine candidates, we expressed a triple mutant of staphylococcal enterotoxin B carrying the L45R, Y89A, and Y94A modifications in transgenic soybean seeds (soy-mSEB). Soy-mSEB possessed no detectable superantigen activity in vitro. We found that this soybean-derived, nontoxic mutant of SEB could be stably expressed, stored in seeds for extended periods at room temperature without degradation, and easily purified from contaminating soy proteins. Vaccination of pigs with purified soy-mSEB, or the identical triple mutant expressed in Escherichia coli (E. coli-mSEB), resulted in high antibody titers against the native toxin in immunized animals. In fact, titers were indistinguishable regardless of the immunogen used, demonstrating the equivalence of soy-mSEB and E. coli-mSEB vaccinations. Antisera from either immunized group were able to block native SEB superantigen activity in an in vitro neutralization assay. Similar results were obtained when immunized animals were challenged with a sublethal dose of native toxin. Significant reductions in toxin-induced serum cytokine levels were observed in soy-mSEB- and E. coli-mSEB-immunized pigs compared to control animals. The reductions in SEB-induced cytokine responses were similar regardless of the immunogen used for vaccination. Surprisingly, however, some clinical symptoms, such as prostration, lethargy, emesis, and/or diarrhea, were still observed in all immunized animals. These studies demonstrate the potential for soybean-derived proteins as a platform technology for sustainable vaccine manufacturing and the usefulness of a sublethal challenge model in pigs for evaluating the efficacy of potential SEB vaccine candidates.
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.
Satellite cell activity is necessary for postnatal skeletal muscle growth. Severe phosphate (PO4) deficiency can alter satellite cell activity, however the role of neonatal PO4 nutrition on satellite cell biology remains obscure. Twenty-one piglets (1 day of age, 1.8 ± 0.2 kg BW) were pair-fed liquid diets that were either PO4 adequate (0.9% total P), supra-adequate (1.2% total P) in PO4 requirement or deficient (0.7% total P) in PO4 content for 12 days. Body weight was recorded daily and blood samples collected every 6 days. At day 12, pigs were orally dosed with BrdU and 12 h later, satellite cells were isolated. Satellite cells were also cultured in vitro for 7 days to determine if PO4 nutrition alters their ability to proceed through their myogenic lineage. Dietary PO4 deficiency resulted in reduced (P < 0.05) sera PO4 and parathyroid hormone (PTH) concentrations, while supra-adequate dietary PO4 improved (P < 0.05) feed conversion efficiency as compared to the PO4 adequate group. In vivo satellite cell proliferation was reduced (P < 0.05) among the PO4 deficient pigs, and these cells had altered in vitro expression of markers of myogenic progression. Further work to better understand early nutritional programming of satellite cells and the potential benefits of emphasizing early PO4 nutrition for future lean growth potential is warranted.
Mesenchymal stem cells (MSC) provide the cells responsible for bone formation, but can also adopt an adipocytic lineage. The balance between osteogenic versus adipocytic differentiation of MSC is of critical importance to bone health. The impact of Ca nutrition on MSC activity was assessed by examining the effect of the Ca status of the MSC and the Ca status of homologous sera, under both proliferative and adipogenic culture conditions. MSC were isolated from 6, 19d old pigs that had been fed either a Ca adequate (Ca+) or deficient (Ca−) milk‐replacer for 18d. Sera were pooled samples from days 6, 12, and 18 of the feeding trial. Cells were cultured for 6d prior to analysis. Under proliferative culture conditions, Ca− sera reduced MSC proliferation and Ca+ MSC had greater expression of osteocalcin and Runx2 mRNA. Under adipogenic culture conditions, MSC cultured in Ca− sera had greater Oil Red O staining and Ca+ MSC also had greater Oil Red O staining than did there Ca− counterparts. Ca− MSC cultured with Ca− sera under adipogenic conditions had 2 fold greater expression of PPARG2 mRNA than any other treatment group. Concentrations of calcitropic hormones were not different between Ca+ and Ca− sera, but we identified 22 differentially expressed proteins in these sera. The results indicate that neonatal Ca restriction may have long‐term effects on bone integrity via programming of MSC.
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