Quantitative Proteomics Analysis of Barley-Based Liquid Feed and the Effect of Protease Inhibitors and NADPH-Dependent Thioredoxin Reductase/Thioredoxin (NTR/Trx) System.

Liquid feeding strategies have been devised with the aim of enhancing grain nutrient availability for livestock. It is characterized by a steeping/soaking period that softens the grains and initiates mobilization of seed storage reserves. The present study uses 2D gel-based proteomics to investigate the role of proteolysis and reduction by thioredoxins over a 48 h steeping period by monitoring protein abundance dynamics in barley-based liquid feed samples supplemented with either protease inhibitors or NADPH-dependent thioredoxin reductase/thioredoxin (NTR/Trx). Several full-length storage proteins were only identified in the water-extractable fraction of feed containing protease inhibitors, illustrating significant inhibition of proteolytic activities arising during the steeping period. Application of functional NTR/Trx to liquid feed reductively increased the solubility of known and potentially new Trx-target proteins, e.g., outer membrane protein X, and their susceptibility to proteolysis. Thus, the NTR/Trx system exhibits important potential as a feed additive to enhance nutrient digestibility in monogastric animals.

Mammary metabolism and colostrogenesis in sows during late gestation and the colostral period.

The aims of this study were to investigate 1) the effect of high dietary fiber (DF; 19.3% to 21.7%) supplemented to late gestating sows on mammary uptake and metabolism of energy substrates as well as colostrum production and 2) the ontogeny of colostral fat and lactose synthesis using mammary carbon balance, and colostral protein using IgG as a biomarker. Sows were fed either a control diet (CON) consisting of a standard gestation diet (14.6% DF) until day 108 of gestation and a transition diet (16.8% DF) from day 109 of gestation until farrowing or a high DF treatment where part of the daily ration was replaced with a high DF supplement (FIB). The FIB sows received 19.3% and 21.7% DF in the last 2 wk prior to farrowing. Sows were surgically implanted with permanent indwelling catheters at day 75 ± 2 of gestation and blood samples were collected at 6 different time points in late gestation and at 11 different time points within 24 h after the onset of farrowing. Colostrum samples were collected at 0, 12, and 24 h after the onset of farrowing. Arterial concentration of acetate (P = 0.05) and colostral fat content (P = 0.009) were greater in FIB sows compared with CON sows. Plasma IgG dropped from day -10 relative to farrowing (P < 0.001), suggesting an uptake by the mammary glands. Mammary plasma flow (P = 0.007) and net mammary uptake of glucose (P = 0.04) increased during farrowing while dietary treatment had no effect on net mammary uptake of other energy substrates during late gestation and farrowing. The net mammary uptake of carbon from glucogenic precursors did not equate to the sum of carbons secreted in colostral lactose and released as CO2, indicating that carbons from ketogenic precursors were likely used for colostral fat and for oxidation. Mammary nonprotein carbon uptake matched the mammary output, indicating that the majority of colostral fat and lactose were produced after the onset of farrowing. In conclusion, high DF included in the diet for late gestating sows increased colostral fat content by 49% but this substantial dietary response could not be explained by the increased carbon uptake from short chain fatty acids during the colostral period. The nonprotein carbon balance of mammary glands during farrowing suggests that the majority of colostral fat and lactose were produced after the onset of farrowing, whereas the drop in plasma IgG in late gestation suggests that the mammary glands take up this colostral component prior to farrowing.

Influence of zinc supplementation on immune parameters in weaned pigs.

Zinc is an essential trace element, highly important for a well functioning immune system. In case of zinc deficiency, proper immune functions are not ensured thus leading to various diseases. Weaning of pigs from the sow causes stress, increasing susceptibility to infections. Moreover, low feed intake during the first two weeks post-weaning, accompanied by low zinc intake, results in temporary zinc deficiency. Therefore, supporting the immune system by zinc supplementation might improve its function and thereby the pigs' health and well-being. In this study, the immune status of weaned pigs was analyzed under different conditions of zinc supplementation. More precisely, the daily porcine diet was either left unsupplemented (0 ppm), or was supplemented with low (100 ppm), or high (2500 ppm) amounts of additional zinc in the form of zinc oxide (ZnO) (Zn0, Zn100, and Zn2500, respectively). Porcine innate and adaptive immune cells of the different dietary groups were analyzed. Results revealed an improved innate immune capacity, represented by increased phagocytosis and slightly increased oxidative burst in cells from the Zn2500 pigs and Zn100 pigs, respectively. Apart from that, zinc supplementation improved adaptive immunity, as seen by increased numbers of CD3+ T cells as well as increased numbers of CD3+CD4+Foxp3+ regulatory T cells, elevated interleukin (IL)-2 production and decreased IL-10 production. Although not significant, supplementing 2500 ppm zinc slightly decreased killing activity of natural killer (NK) cells. Thus, the optimal concentration for zinc supplementation of weaned pigs two weeks post-weaning needs to be further studied, presumably establishing an optimal concentration between 100 ppm and 2500 ppm zinc. Genome comparisons indicate that the porcine genome is more closely related to the human genome than the murine genome is related to the human genome. Therefore, the pig seems to be a suitable organism to study human immunity and diseases. Results obtained in the current study might therefore be transferable to the human immune system.

Corn-Soy-Blend Fortified with Phosphorus to Prevent Refeeding Hypophosphatemia in Undernourished Piglets.

BACKGROUND: Phosphorus (P) levels in refeeding diets are very important as undernourished children are at risk of hypophosphatemia during refeeding. For this reason, conventional corn-soy-blends (CSB) have been reformulated by the World Food Programme to obtain a mono-calcium-phosphate fortified product (CSB+) and a product further fortified with skim milk powder (CBS++). METHODS: Using a piglet model of undernourished children, we hypothesized that feeding of CSB+, CSB++ or CSB+ with added whey permeate (CSB+/wp) would help to prevent refeeding hypophosphatemia. Pigs were weaned at 4 weeks of age and undernutrition was induced with a nutritionally inadequate pure maize diet for 7 weeks, after which they were refed for 3 weeks with either CSB+ (n = 10), CSB++ (n = 10) or CSB+/wp (n = 10). For reference, a fourth group continued on the maize diet (REF, n = 10). RESULTS: Following induction of undernutrition, body weight and length were 29±5% and 67±4% (means±SD) of values in age-matched pigs fed a nutritionally adequate diet, and the mean serum P level was 1.77±0.34 mmol/l. During the first week of refeeding, P levels in the CSB+ pigs decreased to 55% of values before refeeding (P < 0.05) while values in the CSB++ and CSB+/wp pigs were able to maintain their plasma phosphate at a similar level as before refeeding. CONCLUSION: We conclude that fortification of CSB with only monocalcium-phosphate does not prevent hypophosphatemia. Dairy products like skim milk powder or whey permeate may represent relevant sources of phosphorus during refeeding. The content and form of phosphorus in such diets need to be carefully evaluated, and the undernourished piglet may be used to test the efficacy of such diets.

Zinc attenuates forskolin-stimulated electrolyte secretion without involvement of the enteric nervous system in small intestinal epithelium from weaned piglets.

In a previous study, we found that secretagogue-stimulated electrolyte secretion was attenuated by dietary and serosal zinc in piglet small intestinal epithelium in Ussing chambers. Several studies show that the enteric nervous system (ENS) is involved in regulation of electrolyte and/or fluid transport in intestinal epithelium from many species. The aim of the present study is to examine the mechanisms behind the attenuating effect of zinc on electrolyte secretion and to study whether the ENS is involved in this effect of zinc in vitro. Twenty-four piglets (six litters of four piglets) were allocated randomly to one of two dietary treatments consisting of a basic diet supplemented with 100 mg zinc/kg (Zn(100)) or 2500 mg zinc/kg (Zn(2500)), as ZnO. All the piglets were killed at 5-6 days after weaning and in vitro experiments with small intestinal epithelium in Ussing chambers were carried out. Furthermore, zinc, copper, alkaline phosphatase (AP) and metallothionein (MT) in mucosa, liver, and plasma were measured. These measurements showed that zinc status was increased in the Zn(2500) compared to the Zn(100) fed piglets. The in vitro studies did not confirm previous findings of attenuating effects of dietary zinc and zinc in vitro on the 5-HT induced secretion. But it showed that the addition of zinc at the serosal side attenuated the forskolin (FSK) (cAMP-dependent) induced ion secretion in epithelium from piglets fed with Zn(100) diet. Blocking the ENS with lidocaine or hexamethonium apparently slightly reduced this effect of zinc in vitro, but did not remove the effect of zinc. Consequently, it is suggested that zinc attenuates the cAMP dependent ion secretion mainly due to an effect on epithelial cells rather than affecting the mucosal neuronal pathway.

Influence of weaning and effect of post weaning dietary zinc and copper on electrophysiological response to glucose, theophylline and 5-HT in piglet small intestinal mucosa.

This study aimed to examine how weaning and how dietary zinc and/or copper fed post weaning may affect the electrophysiological response to glucose and to chloride secretagogues in piglet small intestine in vitro. Study 1 included 54 piglets (six litters of nine piglets). One piglet from every litter was killed 1 day before weaning. The remaining 48 piglets were allocated at weaning (28 d) to four dietary zinc treatments and subsequently killed 1-2, 5-6 or 14-15 days after weaning. Study 2 included 48 piglets (six litters of eight piglets) allocated to four dietary treatments, consisting of high or low dietary zinc with or without high dietary copper. All piglets in study 2 were killed 5-7 days after weaning. The in vitro studies in Ussing chambers showed that weaning resulted in increased ileal glucose absorption as well as increased neuroendocrine-regulated (activated by 5-HT) and cAMP-dependent (activated by theophylline) chloride secretion. High zinc supplementation reduced the responses to 5-HT and theophylline. The study did not reveal any influence of copper on these parameters. It is concluded that the positive effect of zinc supplementation on diarrhoea in weaned piglets may be due to zinc reducing the intestinal mucosal susceptibility to secretagogues that activate chloride secretion.