Determination of the Optimal Level of Dietary Zinc for Newly Weaned Pigs: A Dose-Response Study.

One hundred and eighty individually housed piglets with an initial body weight of 7.63 ± 0.98 kg (at 28 days of age) were fed a diet containing either 153, 493, 1022, 1601, 2052 or 2407 mg zinc/kg (added Zn as zinc oxide; ZnO) from day 0-21 post weaning to determine the optimal level of Zn for weaned piglets. Body weight, feed intake and faecal scores were recorded, and blood and faecal samples were collected. Dietary Zn content quadratically affected both feed intake and gain in the first two weeks, with an approximately 1400 mg Zn/kg diet and a Zn intake of 400 mg/day as the optimal levels. The relative risk of diarrhoea increased up to 60% at day 7 and 14 if serum Zn status dropped below the weaning level (767 µg/L), and maintain the weaning serum Zn status required approximately 1100 mg Zn/kg (166 mg Zn/day) during week 1. Blood markers of intestinal integrity (D-lactate and diamine oxidase) were unaffected by dietary Zn, and dietary Zn levels of 1022 and 1601 mg/kg did not affect the faecal numbers of total bacteria, Lactobacilli and E. Coli bacteria compared to 153 mg Zn/kg. These results indicate that the requirement for Zn in newly weaned piglets may be substantially higher than currently assumed.

Some Insect Species Are Good-Quality Protein Sources for Children and Adults: Digestible Indispensable Amino Acid Score (DIAAS) Determined in Growing Pigs.

BACKGROUND: Insect species are suitable for farming as "mini livestock" for human and animal consumption. It is important to assess the protein quality of relevant species to understand the potential of these novel protein sources in future sustainable food systems. OBJECTIVES: We aimed to determine the protein quality of 5 insect species-lesser mealworm (LMW), yellow mealworm, house cricket (HC), banded cricket (BC), and black soldier fly-using the digestible indispensable amino acid score (DIAAS) in a pig model. METHODS: Five diets were formulated to contain 10% insect crude protein (CP). A nitrogen (N)-free diet was included to estimate endogenous losses. In a 6 × 6 Latin square design, 6 ileal cannulated crossbred [Duroc × (Danish Landrace × Yorkshire)] male pigs with an initial body weight of 35 ± 2 kg were fed the 6 diets. Each diet was fed for 1 wk over 6 wk. Ileal digesta were collected for 8 h on days 5 and 7 each week. Analyzed CP, amino acid (AA) contents, and calculated values of standardized ileal digestibility for CP and AAs were used to assess the DIAAS of each insect. RESULTS: The DIAAS for young children aged 6 mo-3 y and for older children, adolescents, and adults identified sulfur AAs (cysteine + methionine) as the first limiting AA in all 4 species of cricket and mealworm. For young children, both cricket species had DIAASs > 75 and for older children, adolescents, and adults both cricket species and LMW had DIAASs > 75. CONCLUSIONS: Both cricket species (HC and BC) are classified as good-quality protein sources for young children aged 6 mo-3 y and for older children, adolescents, and adults. One mealworm species, LMW, is a good-quality protein source for older children, adolescents, and adults.

Some Insect Species Are Good-Quality Protein Sources for Children and Adults: Digestible Indispensable Amino Acid Score (DIAAS) Determined in Growing Pigs.

BACKGROUND: Insect species are suitable for farming as "mini livestock" for human and animal consumption. It is important to assess the protein quality of relevant species to understand the potential of these novel protein sources in future sustainable food systems. OBJECTIVES: We aimed to determine the protein quality of 5 insect species-lesser mealworm (LMW), yellow mealworm, house cricket (HC), banded cricket (BC), and black soldier fly-using the digestible indispensable amino acid score (DIAAS) in a pig model. METHODS: Five diets were formulated to contain 10% insect crude protein (CP). A nitrogen (N)-free diet was included to estimate endogenous losses. In a 6 × 6 Latin square design, 6 ileal cannulated crossbred [Duroc × (Danish Landrace × Yorkshire)] male pigs with an initial body weight of 35 ± 2 kg were fed the 6 diets. Each diet was fed for 1 wk over 6 wk. Ileal digesta were collected for 8 h on days 5 and 7 each week. Analyzed CP, amino acid (AA) contents, and calculated values of standardized ileal digestibility for CP and AAs were used to assess the DIAAS of each insect. RESULTS: The DIAAS for young children aged 6 mo-3 y and for older children, adolescents, and adults identified sulfur AAs (cysteine + methionine) as the first limiting AA in all 4 species of cricket and mealworm. For young children, both cricket species had DIAASs > 75 and for older children, adolescents, and adults both cricket species and LMW had DIAASs > 75. CONCLUSIONS: Both cricket species (HC and BC) are classified as good-quality protein sources for young children aged 6 mo-3 y and for older children, adolescents, and adults. One mealworm species, LMW, is a good-quality protein source for older children, adolescents, and adults.

Amino Acid Absorption Profiles in Growing Pigs Fed Different Protein Sources.

The aim of the present study was to determine postprandial amino acid (AA) appearance in the blood of growing pigs as influenced by protein source. Seven growing pigs (average body weight 18 kg), in a 7 × 5 Youden square design, were fitted with a jugular vein catheter and fed seven diets containing wheat, soybean meal, enzyme-treated soybean meal, hydrothermally-treated rapeseed meal, casein, hydrolyzed casein, and a crystalline AA blend with the same AA profile as casein. The latter was not eaten by the pigs, therefore being excluded. Blood samples were collected at -30, 30, 60, 90, 120, 180, and 360 min after a meal and analyzed for free AA. Overall, plasma AA concentrations were highest 60 min after feeding. There were no differences in plasma AA concentration between casein and hydrolyzed casein, but soybean meal resulted in lower AA plasma concentrations compared with enzyme-treated soybean meal at 60 and 120 min after feeding. There were no differences between hydrothermally-treated rapeseed meal and soybean meal. In conclusion, the ingredients could not clearly be categorized as being slow or fast protein with regard to protein digestion and absorption of AA, but soybean meal resulted in a prolonged appearance of plasma AA compared to casein and hydrolyzed casein.

Development of Bacillus subtilis mutants to produce tryptophan in pigs.

OBJECTIVES: To generate tryptophan-overproducing Bacillus subtilis strains for in situ use in pigs, to reduce the feed cost for farmers and nitrogen pollution. RESULTS: A novel concept has been investigated-to generate B. subtilis strains able to produce tryptophan (Trp) in situ in pigs. Mutagenesis by UV was combined with selection on Trp and purine analogues in an iterative process. Two mutants from different wild types were obtained, mutant 1 (M1) produced 1 mg Trp/l and mutant 2 (M2) 14 mg Trp/l. Genome sequence analysis revealed that M1 had three single nuclear polymorphisms (SNPs) and M2 had two SNPs compared to the wild type strains. In both mutants SNPs were found in genes regulating tryptophan synthesis. Reverse transcription PCR confirmed up-regulation of the tryptophan synthesis genes in both mutants, the expression was up to 3 times higher in M2 than in M1. CONCLUSIONS: Tryptophan-excreting B. subtilis strains were obtained with UV-mutagenesis and analogue selection and can be used in animal feed applications.

Nontargeted LC-MS Metabolomics Approach for Metabolic Profiling of Plasma and Urine from Pigs Fed Branched Chain Amino Acids for Maximum Growth Performance.

The metabolic response in plasma and urine of pigs when feeding an optimum level of branched chain amino acids (BCAAs) for best growth performance is unknown. The objective of the current study was to identify the metabolic phenotype associated with the BCAAs intake level that could be linked to the animal growth performance. Three dose-response studies were carried out to collect blood and urine samples from pigs fed increasing levels of Ile, Val, or Leu followed by a nontargeted LC-MS approach to characterize the metabolic profile of biofluids when dietary BCAAs are optimum for animal growth. Results showed that concentrations of plasma hypoxanthine and tyrosine (Tyr) were higher while concentrations of glycocholic acid, tauroursodeoxycholic acid, and taurocholic acid were lower when the dietary Ile was optimum. Plasma 3-methyl-2-oxovaleric acid and creatine were lower when dietary Leu was optimum. The optimum dietary Leu resulted in increased urinary excretion of ascorbic acid and choline and relatively decreased excretion of 2-aminoadipic acid, acetyl-dl-valine, Ile, 2-methylbutyrylglycine, and Tyr. In conclusion, plasma glycocholic acid and taurocholic acid were discriminating metabolites to the optimum dietary Ile. The optimum dietary Leu was associated with reduced plasma creatine and urinary 2-aminoadipic acid and elevated urinary excretion of ascorbic acid and choline. The optimum dietary Val had a less pronounced metabolic response reflected in plasma or urine than other BCAA.

Effects of Acidifying Pig Diets on Emissions of Ammonia, Methane, and Sulfur from Slurry during Storage.

Ammonia (NH) volatilization from intensive livestock production is a threat to natural ecosystems. This study investigated pig diet manipulation by 1% (w/w) benzoic acid (BA) amendment and lowering of dietary electrolyte balance through substituting 1.4% (w/w) CaCO with 2.0% (w/w) CaCl. Urine and feces were collected separately from 24 pigs fed one of four diets (Control, +BA, +CaCl, +BA+CaCl) in metabolic cages and mixed as slurry. During 103 d of storage, all acidifying diets consistently reduced pH in the slurry by 0.4 to 0.6 units. There was a strong relationship between slurry pH and NH emissions, which were considerably reduced by the three acidifying diets. The +BA diet decreased NH emission by 28%, the +CaCl diet by 37%, and the combined +BA and +CaCl diet by 40%. Acidifying diets had no effect on S cycling or emission of volatile S compounds under the prevailing conditions of restricted S feeding. Methane (CH) emissions were increased by 73% in diets with CaCl. An initial delay in CH emissions was investigated in a separate experiment with manipulation of pH (5.4, 6.7, or 8.8) and inoculation with adapted pig slurry (0, 4, 11, or 19%), which showed that methanogenic potential, rather than inhibitory effects of the chemical environment, caused the delay. In conclusion, NH emissions from slurry could be reduced by addition of BA to pig diets or by controlling the dietary electrolyte balance, but there was no additive effect of combining the two strategies. However, CH emissions from slurry may increase with acidifying diets.

Emissions of sulfur-containing odorants, ammonia, and methane from pig slurry: effects of dietary methionine and benzoic acid.

Supplementation of benzoic acid to pig diets reduces the pH of urine and may thereby affect emissions of ammonia and other gases from slurry, including sulfur-containing compounds that are expected to play a role in odor emission. Over a period of 112 d, we investigated hydrogen sulfide (H(2)S), methanethiol (MT), dimethyl sulfide (DMS), dimethyl disulfide (DMDS), and dimethyl trisulfide (DMTS), as well as ammonia and methane emissions from stored pig slurry. The slurry was derived from a feeding experiment with four pig diets in a factorial design with 2% (w/w) benzoic acid and 1% (w/w) methionine supplementation as treatments. Benzoic acid reduced slurry pH by 1 to 1.5 units and ammonia emissions by 60 to 70% for up to 2 mo of storage, and a considerable, but transitory reduction of methane emissions was also observed after 4 to 5 wk. All five volatile sulfur (S) compounds were identified in gas emitted from the slurry of the control treatment, which came from pigs fed according to Danish recommendations for amino acids and minerals. The emission patterns of volatile S compounds suggested an intense cycling between pools of organic S in the slurries, with urinary sulfate as the main source. Diet supplementation with methionine significantly increased all S emissions. Diet supplementation with benzoic acid reduced emissions of H(2)S and DMTS compared with the control slurry and moderately increased the concentrations of MT. Sulfur gas emissions were influenced by a strong interaction between methionine and benzoic acid treatments, which caused a significant increase in emissions of especially MT, but also of DMDS. In conclusion, addition of 2% benzoic acid to pig diets effectively reduced ammonia volatilization, but interactions with dietary S may increase odor problems.

Estrogenic activity of bovine milk high or low in equol using immature mouse uterotrophic responses and an estrogen receptor transactivation assay.

BACKGROUND: Cow's milk contain phytoestrogens especially equol depending on the composition of the feed ration. However, it is unknown whether milk differing in equol exhibits different estrogenicity in model systems and thereby potentially in humans as milk consumers. METHODS: The estrogenicity of high and low equol milk (HEM and LEM, respectively) and purified equol was investigated in immature female mice including mRNA expression of six estrogen-sensitive genes in uterine tissue. Extracts of HEM and LEM were also tested for estrogenicity in vitro in an estrogen receptor (ER) reporter gene assay with MVLN cells. RESULTS: The total content of phytoestrogens was approximately 10 times higher in HEM compared with LEM, but levels of endogenous milk estrone and 17beta-estradiol were similar in the two milk types (503-566 and 60-64.6pg/ml, respectively). There was no difference in uterine weight between mice receiving LEM and HEM, and no difference from controls. Equol (50 times the concentration in HEM) was not uterotrophic. The ERbeta mRNA expression was down-regulated in the uteri of HEM mice compared with LEM and controls, but there was no difference between milk types for any of the other genes. Extracts of HEM showed a higher estrogenicity than extracts of LEM in MVLN cells, and there was a dose-dependent increase in estrogenicity by equol. CONCLUSION: The higher in vitro estrogenicity of HEM was not reflected as a higher uterine weight in vivo although the down-regulation of ERbeta in uterine tissue of HEM mice could suggest some estrogenic activity of HEM at the gene expression level.

Bioavailability of alpha-tocopherol stereoisomers in rats depends on dietary doses of all-rac- or RRR-alpha-tocopheryl acetate.

The biological function of the stereoisomers of alpha-tocopherol is believed to depend on their bioavailability. Assessment of bioavailability within the body is therefore considered to be a good and easy way to predict biological value. The separation of alpha-tocopherol methyl ethers by chiral column HPLC is a good and easy tool with which to study the distribution of alpha-tocopherol stereoisomers. The objective of this investigation was to evaluate the bioavailability and distribution of the stereoisomers of alpha-tocopherol in the plasma and tissue in growing rats fed 25, 50, 100 or 200 mg/kg diet of either RRR- or all-rac-alpha-tocopheryl acetate for 10 d. The ratio between the two vitamin E sources based on their alpha-tocopherol concentration in plasma and tissues varied in the plasma between 1.04 and 1.74 and in tissues, ratios of 0.84-1.24 for liver, 0.34-1.59 for lung and 0.75-1.50 for spleen were obtained. An increasing dietary level of all-rac-alpha-tocopheryl acetate decreased the proportion of RRR-alpha-tocopherol, whereas the other stereoisomers were not affected. RRS-alpha-Tocopherol was present in the highest proportion, followed by RSR-, RSS- and RRR-alpha-tocopherol. In contrast to the other tissues and plasma, the liver contained the highest proportion (29-33 %) of the four 2S stereoisomers of total alpha-tocopherol. Rats fed RRR-alpha-tocopheryl acetate for 10 d showed a significant increase in the plasma and tissue content of RRR-alpha-tocopherol and a simultaneous decrease in the other three 2R isomers, whereas the absolute content of the 2S isomers was unaffected. In adipose tissue, concentrations of the three synthetic 2R isomers remained constant, whereas there was a steep increase in the content of RRR-alpha-tocopherol.