Metabolic footprint and intestinal microbial changes in response to dietary proteins in a pig model.

Epidemiological studies revealed that dietary proteins can contribute to the modulation of the cardiovascular disease risk. Still, direct effects of dietary proteins on serum metabolites and other health-modulating factors have not been fully explored. Here, we compared the effects of dietary lupin protein with the effects of beef protein and casein on the serum metabolite profile, cardiovascular risk markers and the fecal microbiome. Pigs were fed diets containing 15% of the respective proteins for 4 weeks. A classification analysis of the serum metabolites revealed six biomarker sets of two metabolites each that discriminated between the intake of lupin protein, lean beef or casein. These biomarker sets included 1- and 3-methylhistidine, betaine, carnitine, homoarginine and methionine. The study revealed differences in the serum levels of the metabolites 1- and 3- methylhistidine, homoarginine, methionine and homocysteine, which are involved in the one-carbon cycle. However, these changes were not associated with differences in the methylation capacity or the histone methylation pattern. With the exception of serum homocysteine and homoarginine levels, other cardiovascular risk markers, such as the homeostatic model assessment index, trimethylamine-N-oxide and lipids, were not influenced by the dietary protein source. However, the composition of the fecal microorganisms was markedly changed by the dietary protein source. Lupin-protein-fed pigs exhibited more species from the phyla Bacteroidetes and Firmicutes than the other two groups. In conclusion, different dietary protein sources induce distinct serum metabolic fingerprints, have an impact on the cardiovascular risk and modulate the composition of the fecal microbiome.

Impact of a high-protein diet during lactation on milk composition and offspring in a pig model.

PURPOSE: Early postnatal nutrition not only holds relevance to infant growth, but also determines the risk of developing obesity and chronic diseases such as diabetes type 2 and cardiovascular diseases in adulthood. It is suggested that a high-protein (HP) diet in early childhood can predispose children to obesity. However, data concerning possible alterations in milk composition and the development of the offspring in response to a maternal HP diet are currently not available. To address this question, we conducted a study using pigs as a model organism. METHODS: At parturition, sows were assigned to two experimental groups. During lactation, the control group received a diet with a protein content of 16%, whereas the diet of the HP group contained 30% protein. After 28 days of lactation, samples were taken from sows and piglets for the quantification of free amino acids and other metabolites and for histology. RESULTS: Serum and milk urea showed the most marked differences between the two groups of sows, whereas serum urea concentration in piglets did not differ. Here, we found that the intake of an HP diet changed a series of metabolites in sows, but had only small effects on milk composition and virtually no effects on growth in the offspring. Interestingly, maternal protein intake during lactation shapes the microbiome of the offspring. CONCLUSION: From our current study, we conclude that even a very high maternal protein intake throughout lactation has no impact on growth and health parameters of the offspring.

Agrimonia procera exerts antimicrobial effects, modulates the expression of defensins and cytokines in colonocytes and increases the immune response in lipopolysaccharide-challenged piglets.

BACKGROUND: Because antibiotic use in livestock is assumed to contribute to the emerging public health crisis of antibiotic resistance, alternatives are required. Phytogenic additives are extensively studied due to their antibiotic properties. Components of Agrimonia species have been reported as candidate antimicrobials that possess antioxidative and anti-inflammatory properties. We studied the impact of Agrimonia procera (AP) on the growth of selected strains of gut bacteria, the effect of AP on the mRNA abundance of genes involved in inflammation and bacterial defense in a colon carcinoma cell line, the effect of AP in piglets challenged with lipopolysaccharides, and the effect of AP on the growth performance of healthy piglets. RESULTS: The in vitro growth rate of different bacteria strains was negatively affected by AP, especially in Pediococcus pentosaceus and all tested E. coli strains. Stimulation of Caco-2 cells with TNFα resulted in elevated mRNA expression of CXCL1, IL-8 and GPX2. After pretreatment of cells with AP, stimulation of Caco-2 cells with TNFα still resulted in elevated mRNA expression of CXCL1 and IL-8 at all measured points in time. However, mRNA expression in AP-pretreated cells was lower after 6 h and 24 h. In addition, expression of DEFB1 and GPX2 was significantly elevated after TNFα stimulation. In vivo, application of lipopolysaccharides induced significantly increased animal body temperatures. Piglets pretreated with AP prior to lipopolysaccharide application showed a faster and larger increase in body temperature than controls. In addition, piglets pretreated with AP appeared to release more TNFα than controls. In healthy piglets, AP treatment had no impact on growth performance parameters. Fecal dry matter and total plasma antioxidant capacity tended to be higher in piglets treated with AP than in control piglets (P = 0.055 and P = 0.087, respectively). CONCLUSIONS: AP has antimicrobial effects in vitro and stimulated the expression of proinflammatory cytokines in Caco-2 cells. The additive had no effect on growth in healthy piglets but increased the immune response in LPS-treated animals. In addition, AP appeared to have antioxidative effects in vivo. Therefore, AP merits testing as a future alternative to antibiotics in animal husbandry.

High Leucine Diets Stimulate Cerebral Branched-Chain Amino Acid Degradation and Modify Serotonin and Ketone Body Concentrations in a Pig Model.

In addition to its role as an essential protein component, leucine (Leu) displays several other metabolic functions such as activation of protein synthesis. This property makes it an interesting amino acid for the therapy of human muscle atrophy and for livestock production. However, Leu can stimulate its own degradation via the branched-chain keto acid dehydrogenase complex (BCKDH). To examine the response of several tissues to excessive Leu, pigs were fed diets containing two- (L2) and four-fold (L4) higher Leu contents than the recommended amount (control). We found that the L4 diet led to a pronounced increase in BCKDH activity in the brain (2.5-fold, P < 0.05), liver (1.8-fold, P < 0.05) and cardiac muscle (1.7-fold, P < 0.05), whereas we found no changes in enzyme activity in the pancreas, skeletal muscle, adipose tissue and intestinal mucosa. The L2 diet had only weak effects on BCKDH activity. Both high Leu diets reduced the concentrations of free valine and isoleucine in nearly all tissues. In the brain, high Leu diets modified the amount of tryptophan available: for serotonin synthesis. Compared to the controls, pigs treated with the high Leu diets consumed less food, showed increased plasma concentrations of 3-hydroxybutyrate and reduced levels of circulating serotonin. In conclusion, excessive Leu can stimulate BCKDH activity in several tissues, including the brain. Changes in cerebral tryptophan, along with the changes in amino acid-derived metabolites in the plasma may limit the use of high Leu diets to treat muscle atrophy or to increase muscle growth.

The phytochemical investigation of Agrimonia eupatoria L. and Agrimonia procera Wallr. as valid sources of Agrimoniae herba--The pharmacopoeial plant material.

The agrimony herb is a traditional plant drug, which is commonly used as a mildly astringent agent. According to European Pharmacopoeia, the only source of this plant drug is Agrimonia eupatoria. By contrast the German Commission E pharmacopoeial monograph used to allow Agrimonia procera to be used as a second valid source of Agrimoniae herba. Several studies have been conducted on the phytochemical composition of common agrimony. The data on the phytochemistry of A. procera are scarce. The aim of the present study was an in-depth phytochemical comparison of A. eupatoria and A. procera in the context of the pharmacopoeial monograph of A. herba. The comparison of two agrimony species showed that there are no significant qualitative differences. The quantitative HPLC analysis revealed that fragrant agrimony is a much better source of agrimoniin than common agrimony. This difference could not be detected using the pharmacopoeial method of quantification for the total tannin content. The present study has shown for the first time the possible use of apigenin-C-glycosides (vitexin and isovitexin) as chemotaxonomic markers for distinguishing both agrimony species. The potential chemical markers such as apigenin-7-O-glucoside and high agrimoniin content were also suggested for fragrant agrimony. Based on the data obtained, A. procera should be considered as a valid source of pharmacopoeial plant material.

Studies on the health impact of Agrimonia procera in piglets.

BACKGROUND: The weaning period is critical for stress-related diseases and infections. Currently, large amounts of therapeutic antimicrobials are used to treat infections in the livestock production, especially in piglets. Phytogenic feed additives could provide a useful alternative. We hypothesize, that components in agrimonia species which have been used successfully in humans to treat gastrointestinal infections could also improve the health of piglets. We investigated the effects of Agrimonia procera (AP) on the growth performance of piglets and cytokine expression in isolated porcine peripheral blood mononuclear cells (PBMC). RESULTS: Here we show that piglets that received a diet with 0.56 g/kg AP for 6 weeks tended to ingest more food (+5.1%; P < 0.10), and were characterized by a higher nitrogen retention (+9.6%, P < 0.05) than the control group without AP treatment. Data from a second experiment reveal that piglets fed a diet with 0.87 g/kg AP for 6 weeks had an improved food conversion ratio (1.46 ± 0.04) compared to those that received none (1.54 ± 0.08) or 8.7 g/kg AP (1.60 ± 0.08) with their diets (P < 0.001). However, the food intake, daily weight gain and dry matter of feces were not affected by the AP treatment. Treatment of PBMC for 1 and 6 h with AP extract (APE) reduced the mRNA abundance of tumor necrosis factor (TNF)? in cells challenged with lipopolysaccharides (LPS) but not in cells without LPS stimulation (P < 0.05). The lower mRNA expression of TNF? was accompanied by a trend towards a lower release of TNF? from these cells (P?=?0.067). After the treatment of PBMC with APE for 6 h, the relative mRNA concentration of interleukin (IL)-1? declined (P < 0.05), whereas that of IL-10 remained unchanged. Treatment of LPS-challenged PBMC for 20 h with varying concentrations of APE did not reveal any effect on cytokine expression and TNF? release. CONCLUSIONS: The results indicate that low dosages of AP may improve the growth performance of piglets and seem to exert antiinflammatory effects in porcine immune cells challenged with LPS.

Free-range farming: a natural alternative to produce vitamin D-enriched eggs.

OBJECTIVE: Food-based strategies need to be developed to improve the vitamin D status of individuals. Recent studies identified ultraviolet B irradiation as an efficient method to enrich mushrooms and eggs with vitamin D. The aim of this study was to determine whether free-range farming of hens could provide a valuable method to produce vitamin D-enriched eggs. METHODS: Laying hens were randomly assigned to three groups of 33 to 34 animals each, and were kept either indoors (indoor group), outdoors (outdoor group), or with an indoor/outdoor option (indoor/outdoor group) over 4 wk. RESULTS: The study shows that the vitamin D3 content of egg yolk was three- to fourfold higher in the groups that were exposed to sunlight (outdoor and indoor/outdoor groups) compared with the indoor group (P < 0.001). Egg yolk from the outdoor group revealed the highest vitamin D3 content, which averaged 14.3 µg/100 g dry matter (DM), followed by that from the indoor/outdoor group (11.3 µg/100 g DM). Yolk from indoor eggs contained only 3.8 µg vitamin D/100 g DM. The 25-hydroxyvitamin D (25[OH]D3) content of egg yolk was also influenced by sunlight exposure, although less pronounced than the vitamin D content (P < 0.05). In contrast, free-range eggs randomly acquired from supermarkets had relatively low vitamin D contents. CONCLUSION: Free-range farming offers an efficient alternative to fortify eggs with vitamin D, provided that farming conditions are sufficiently attractive for hens to range outside.

Lupin protein isolate versus casein modifies cholesterol excretion and mRNA expression of intestinal sterol transporters in a pig model.

BACKGROUND: Lupin proteins exert hypocholesterolemic effects in man and animals, although the underlying mechanism remains uncertain. Herein we investigated whether lupin proteins compared to casein modulate sterol excretion and mRNA expression of intestinal sterol transporters by use of pigs as an animal model with similar lipid metabolism as humans, and cellular cholesterol-uptake by Caco-2 cells. METHODS: Two groups of pigs were fed cholesterol-containing diets with either 230 g/kg of lupin protein isolate from L. angustifolius or 230 g/kg casein, for 4 weeks. Faeces were collected quantitatively over a 5 d period for analysis of neutral sterols and bile acids by gas chromatographically methods. The mRNA abundances of intestinal lipid transporters were analysed by real-time RT-PCR. Cholesterol-uptake studies were performed with Caco-2 cells that were incubated with lupin conglutin γ, phytate, ezetimibe or albumin in the presence of labelled [4-14C]-cholesterol. RESULTS: Pigs fed the lupin protein isolate revealed lower cholesterol concentrations in total plasma, LDL and HDL than pigs fed casein (P < 0.05). Analysis of faeces revealed a higher output of cholesterol in pigs that were fed lupin protein isolate compared to pigs that received casein (+57.1%; P < 0.05). Relative mRNA concentrations of intestinal sterol transporters involved in cholesterol absorption (Niemann-Pick C1-like 1, scavenger receptor class B, type 1) were lower in pigs fed lupin protein isolate than in those who received casein (P < 0.05). In vitro data showed that phytate was capable of reducing the uptake of labelled [4-14C]-cholesterol into the Caco-2 cells to the same extend as ezetimibe when compared to control (-20.5% vs. -21.1%; P < 0.05). CONCLUSIONS: Data reveal that the cholesterol-lowering effect of lupin protein isolate is attributable to an increased faecal output of cholesterol and a reduced intestinal uptake of cholesterol. The findings indicate phytate as a possible biofunctional ingredient of lupin protein isolate.

UVB exposure of farm animals: study on a food-based strategy to bridge the gap between current vitamin D intakes and dietary targets.

Vitamin D deficiency is a global health problem. This study aimed to investigate the efficacy of ultraviolet (UV) B radiation for improving vitamin D3 content of eggs and meat. In a two-factorial design hens that received diets with 0 (-D3) or 3,000 IU (+D3) vitamin D3/kg were non-exposed (-UVB) or exposed to UVB radiation (+UVB) for 3 h daily over 4 weeks. Data show that UVB radiation was very effective in raising the vitamin D3 content of egg yolk and meat. Egg yolk from +UVB/-D3 hens had a higher vitamin D3 content (17.5±7.2 µg/100 g dry matter (DM)) than those from the -UVB/+D3 group (5.2±2.4 µg/100 g DM, p<0.01). Vitamin D3 content in egg yolk of vitamin D3-supplemented hens could be further increased by UVB radiation (32.4±10.9 µg/100 g DM). The content of 25-hydroxyvitamin D3 (25(OH)D3) in the egg yolk also increased in response to UVB, although less pronounced than vitamin D3. Meat revealed about 4-fold higher vitamin D3 contents in response to UVB than to dietary vitamin D3 (p<0.001). In conclusion, exposure of hens to UVB is an efficient approach to provide consumers with vitamin D3-enriched foods from animal sources.

Effects of dietary benzoic acid and sodium-benzoate on performance, nitrogen and mineral balance and hippuric acid excretion of piglets.

The objective of this study was to compare the effects of sodium-benzoate (NaB) with those of benzoic acid (BAc) on growth performance of piglets as well as nutrient digestibility, nitrogen and mineral balance, urinary pH, and the urinary excretion of BAc and hippuric acid (HAc). The study was conducted with 120 weaning piglets (6.5 kg body weight), divided in four groups (15 replicates of two piglets each), which received (1) a basal diet (Control), or the basal diet supplemented with (2) 4 g NaB per kg (Group 4NaB), (3) 3.5 g BAc per kg (Group 3.5BAc) or (4) 5 g BAc per kg (Group 5BAc). Performance data were monitored over a 42-day period. Urine and faeces were collected from day 28-33 in metabolic cages with five piglets per treatment. Piglets of Groups 3.5BAc and 5BAc had similarly a considerably improved average daily gain and feed intake (p < 0.05). Performance of Group 4NaB was not significantly different from the other groups. Compared to the Control, the nitrogen retention was only improved in Group 5BAc (p < 0.05); the other groups showed intermediate values. In the supplemented groups, most of the BAc was excreted as HAc in urine, but only Groups 3.5BAc and 5BAc had reduced urinary pH (p < 0.05). Daily intake and faecal and urinary excretion of P and Ca were not affected by the treatment. The molar excess of Na in Group 4NaB was reflected by higher renal excretion of Na compared to the other groups (p < 0.05).

Effect of short-term UVB exposure on vitamin D concentration of eggs and vitamin D status of laying hens.

Vitamin D deficiency in humans is widespread, and only a few food items are important natural sources of vitamin D. This study investigated the effect of UVB exposure of laying hens on the vitamin D content in egg yolk. In a two-factorial design, hens fed a vitamin D-deficient (-D) or -adequate (+D) diet were nonexposed or exposed to UVB light over a period of 4 weeks. UVB exposure of the -D group caused nearly normal egg production rate and egg shell quality; exposure of the +D group did not further improve these parameters. UVB exposure tended to improve the concentration of plasma 25-hydroxycholecalciferol (25(OH)D(3)), but had no effect on 1,25-dihydroxycholecalciferol in plasma or on cholecalciferol and 25(OH)D(3) in egg yolk. The present study shows that a short-term exposure of laying hens to UVB light is not an appropriate way to improve the vitamin D content of egg yolk.

Influence of broccoli extract and various essential oils on performance and expression of xenobiotic- and antioxidant enzymes in broiler chickens.

The aim of our present study was to examine the regulation of xenobiotic- and antioxidant enzymes by phytogenic feed additives in the intestine and the liver of broilers. A total of 240 male Ross-308 broiler chickens (1 d old) were fed a commercial starter diet for 2 weeks. On day 15, the birds were assigned to six treatment groups of forty birds each. The control (Con) group was fed a diet without any additive for 3 weeks. The diet of group sulforaphane (SFN) contained broccoli extract providing 0.075 g/kg SFN, whereas the diets of the other four groups contained 0.15 g/kg essential oils from turmeric (Cuo), oregano (Oo), thyme and rosemary (Ro). Weight gain and feed conversion were slightly impaired by Cuo and Oo. In the jejunum SFN, Cuo and Ro increased the expression of xenobiotic enzymes (epoxide hydrolases 1 and 2 and aflatoxin B1 aldehyde reductase) and of the antioxidant enzyme haeme oxygenase regulated by an 'antioxidant response element' (ARE) compared to group Con. In contrast to our expectations in the liver, the expression of these enzymes was decreased by all the additives. Nevertheless, all the additives increased the Trolox equivalent antioxidant capacity of the jejunum and the liver and reduced Fe-induced lipid peroxidation in the liver. We conclude that the up-regulation of ARE genes in the small intestine reduces oxidative stress in the organism and represents a novel mechanism by which phytogenic feed additives improve the health of farm animals.

Effect of L-carnitine on the hepatic transcript profile in piglets as animal model.

BACKGROUND: Carnitine has attracted scientific interest due to several health-related effects, like protection against neurodegeneration, mitochondrial decay, and oxidative stress as well as improvement of glucose tolerance and insulin sensitivity. The mechanisms underlying most of the health-related effects of carnitine are largely unknown. METHODS: To gain insight into mechanisms through which carnitine exerts its beneficial metabolic effects, we fed piglets either a control or a carnitine supplemented diet, and analysed the transcriptome in the liver. RESULTS: Transcript profiling revealed 563 genes to be differentially expressed in liver by carnitine supplementation. Clustering analysis of the identified genes revealed that most of the top-ranked annotation term clusters were dealing with metabolic processes. Representative genes of these clusters which were significantly up-regulated by carnitine were involved in cellular fatty acid uptake, fatty acid activation, fatty acid ß-oxidation, glucose uptake, and glycolysis. In contrast, genes involved in gluconeogenesis were down-regulated by carnitine. Moreover, clustering analysis identified genes involved in the insulin signaling cascade to be significantly associated with carnitine supplementation. Furthermore, clustering analysis revealed that biological processes dealing with posttranscriptional RNA processing were significantly associated with carnitine supplementation. CONCLUSION: The data suggest that carnitine supplementation has beneficial effects on lipid and glucose homeostasis by inducing genes involved in fatty acid catabolism and glycolysis and repressing genes involved in gluconeogenesis.

mRNA expression of genes involved in fatty acid utilization in skeletal muscle and white adipose tissues of sows during lactation.

Rodents are able to lower fatty acid utilization in liver and muscle during lactation in order to spare fatty acids for the production of milk triacylglycerols, an effect which is mediated by a down-regulation of peroxisome proliferator-activated receptor α (PPARα). The present study was performed to investigate whether similar fatty acid sparing effects are developing in lactating sows. We considered PPARα and its target genes involved in fatty acid utilization in biopsy samples from muscle and adipose tissue of lactating compared to non-lactating sows. In muscle, PPARα target genes involved in fatty acid utilization were up-regulated during lactation indicating that the fatty acid utilization in muscle was increased. Activation of PPARα was probably due to increased concentrations of non-esterified fatty acids in plasma observed in the lactating sows. In contrast to muscle, PPARα and its target genes involved in ß-oxidation in white adipose tissue were down-regulated in early lactation. Overall, the present study shows that sows, unlike rats, are not able to reduce the fatty acid utilization in muscle in order to spare fatty acids for milk production. However, fatty acid oxidation in adipose tissue is lowered during early lactation, an effect that might be helpful to conserve fatty acids released from adipose tissue for the delivery into other tissues, including mammary gland, via the blood.

Dietary L-carnitine alters gene expression in skeletal muscle of piglets.

SCOPE: Carnitine improves protein accretion, muscle mass, and protein:fat accretion in piglets. The underlying mechanisms, however, are largely unknown. METHODS AND RESULTS: To gain insight into mechanisms through which carnitine exerts these effects, we fed piglets either a control or a carnitine-supplemented diet, and analyzed the transcriptome in skeletal muscle. Carnitine concentrations in plasma and muscle were about four-fold higher in the carnitine group when compared to the control group. Transcript profiling revealed 211 genes to be differentially expressed in muscle by carnitine supplementation. The identified genes were mainly involved in molecular processes such as cytoskeletal protein binding, insulin-like growth factor (IGF) binding, transcription factor activity, and insulin receptor binding. Identified genes with the molecular function transcription factor activity encoded primarily transcription factors, most of which were down-regulated by carnitine, including pro-apoptotic transcription factors such as proto-oncogene c-fos, proto-oncogene c-jun and activating transcription factor 3. Furthermore, atrophy-related genes such as atrogin-1, MuRF1, and DRE1 were significantly down-regulated by carnitine. IGF signalling and insulin signalling were identified as significantly up-regulated regulatory pathways in the carnitine group. CONCLUSION: Carnitine may have beneficial effects on skeletal muscle mass through stimulating the anabolic IGF-1 pathway and suppressing pro-apoptotic and atrophy-related genes, which are involved in apoptosis of muscle fibers and proteolysis of muscle proteins, respectively.

Activities of gamma-butyrobetaine dioxygenase and concentrations of carnitine in tissues of pigs.

In contrast to other species, less is known about carnitine homeostasis in the pig. This study was performed to yield information about the site of carnitine synthesis and carnitine concentrations in various tissues of pigs (Sus scrofa). We found that among several pig tissues, a considerable activity of gamma-butyrobetaine dioxygenase (BBD), the last enzyme of carnitine synthesis, exists, like in humans and several other species, only in liver and kidney. Activity of that enzyme in liver and kidney was lower at birth than in the subsequent weeks of life. Highest carnitine concentrations were found in skeletal muscle and heart. Carnitine concentrations in plasma, liver and kidney at birth were higher than in the subsequent weeks of life in spite of the low BBD activity at birth. In conclusion, this study shows that liver and kidney are the major sites of carnitine synthesis and that neonatal pigs do not have an insufficient carnitine status.

Supplementation of L-carnitine in pigs: absorption of carnitine and effect on plasma and tissue carnitine concentrations.

This study was performed to investigate the bioavailability of carnitine supplements and their effects on the carnitine status of pigs. Seven groups of young pigs with an average body weight of 10 kg were fed a basal diet or the same diets supplemented with 25, 50, 100, 200, 500 or 1000 mg of L-carnitine per kg for 20 days. Absorption rate of the supplemented carnitine in the small intestine, assessed by the use of titanium dioxide as an indigestible indicator, was greater than 95% for the lower doses (25, 50, 100 mg/kg) and greater than 90% for the higher doses (200, 500, 1000 mg/kg). Supplementation of carnitine caused a dose-dependent increase of free carnitine, acetyl and total carnitine concentrations in plasma, liver, kidney, heart and skeletal muscle. At the highest dose of 1000 mg/kg, plasma and tissue total carnitine concentrations were 3- to 6-fold higher than in the unsupplemented control group. In conclusion, the present study shows that young pigs have a high capacity to absorb carnitine from the diet. It is also shown that plasma and tissue carnitine concentrations in young pigs can be markedly increased by supplementation of carnitine.

Effect of xylanase on apparent ileal and total tract digestibility of nutrients and energy of rye in young pigs.

A digestibility experiment was carried out on weanling piglets to study the effect of an enzyme complex with predominant xylanase activity on apparent ileal (AID) and apparent total tract digestibility of nutrients and energy. The enzyme was supplemented at four levels (0, 50, 100 and 200 mg/kg) to a diet containing 96% rye. There were significant effects of the added enzyme on AID of dry matter, organic matter and crude fibre, and on apparent total tract digestibility of dry matter, organic matter and energy. However, the improvements in the digestibility were rather small. Except for galactose, there was a significant response in AID of all non-starch polysaccharide constituents to enzyme supplementation, the greatest effect being found at 100 mg/kg. The improvement in AID of arabinose + xylose (685%) was much higher than that of the remaining sugars (110%). AID of galactose was negative in all dietary treatments, presumably due to its high concentration in endogenous secretions. There was a significant response in AID of the sum of essential and total amino acids to the increased level of the enzyme. It is concluded that the enzyme complex is efficient in degrading dietary fibre components, thus improving the digestibility of organic matter, amino acids and energy.

A moderate excess of dietary lysine lowers plasma and tissue carnitine concentrations in pigs.

This study was performed to investigate whether dietary lysine concentration influences the carnitine status of pigs. Therefore, an experiment with twenty young pigs with an average body weight of 21 kg was performed which were fed either a control diet (9.7 g lysine/kg) or a diet with a moderate excess of lysine (16.8 g lysine/kg). Concentrations of all the other amino acids did not differ between the diets. Pigs fed the high-lysine diet had lower concentrations of free and total carnitine in plasma, liver, kidney and skeletal muscle than control pigs (P<0.05). Pigs fed the high-lysine diet moreover had an increased concentration of trimethyllysine (TML), a reduced mRNA abundance of TML dioxygenase and reduced concentrations of gamma-butyrobetaine (BB) in muscle, indicating that the conversion of TML into BB in muscle was impaired. Concentrations of BB, the metabolic precursor of carnitine, in plasma, liver and kidney were also reduced in pigs fed the high-lysine diet while the activity of BB dioxygenase in kidney was not different and that in liver was even increased compared to control pigs (P<0.05). In conclusion, this study shows that a moderate dietary excess of lysine lowers plasma and tissue carnitine concentrations in pigs. Reduced concentrations of BB in liver and kidney suggest that the depressed carnitine status was likely caused by a decreased rate of carnitine synthesis due to a diminished availability of carnitine precursor, probably mainly as a result of an impaired BB formation in muscle.

Carnitine synthesis and uptake into cells are stimulated by fasting in pigs as a model of nonproliferating species.

In rodents, fasting increases the carnitine concentration in the liver by an up-regulation of enzymes of hepatic carnitine synthesis and novel organic cation transporter (OCTN) 2, mediated by activation of peroxisome proliferator-activated receptor (PPAR) alpha. This study was performed to investigate whether such effects occur also in pigs which like humans, as nonproliferating species, have a lower expression of PPARalpha and are less responsive to treatment with PPARalpha agonists than rodents. An experiment with 20 pigs was performed, which were either fed a diet ad-libitum or fasted for 24 h. Fasted pigs had higher relative mRNA concentrations of the PPARalpha target genes carnitine palmitoyltransferase 1 and acyl-CoA oxidase in liver, heart, kidney, and small intestinal mucosa than control pigs, indicative of PPARalpha activation in these tissues (P<.05). Fasted pigs had a higher activity of gamma-butyrobetaine dioxygenase (BBD), enzyme that catalyses the last step of carnitine biosynthesis in liver and kidney, and higher relative mRNA concentrations of OCTN2, the most important carnitine transporter, in liver, kidney, skeletal muscle, and small intestinal mucosa than control pigs (P<.05). Fasted pigs moreover had higher concentrations of free and total carnitine in liver and kidney than control pigs (P<.05). This study shows for the first time that fasting increases the activity of BBD in liver and kidney and up-regulates the expression of OCTN2 in various tissues of pigs, probably mediated by PPARalpha activation. It is concluded that nonproliferating species are also able to cover their increased demand for carnitine during fasting by an increased carnitine synthesis and uptake into cells.