Gene expression profiling in hepatic tissue of newly weaned pigs fed pharmacological zinc and phytase supplemented diets.
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ABSTRACT:
SUBMITTER: Martinez-Montemayor MM
PROVIDER: S-EPMC2566318 | biostudies-literature | 2008
REPOSITORIES: biostudies-literature
Publications
Gene expression profiling in hepatic tissue of newly weaned pigs fed pharmacological zinc and phytase supplemented diets.
Martínez-Montemayor Michelle M MM Hill Gretchen M GM Raney Nancy E NE Rilington Valencia D VD Tempelman Robert J RJ Link Jane E JE Wilkinson Christopher P CP Ramos Antonio M AM Ernst Catherine W CW
BMC genomics 20080917
<h4>Background</h4>Zinc (Zn) is an essential trace element. However, Zn bioavailability from commonly consumed plants may be reduced due to phytic acid. Zn supplementation has been used to treat diarrheal disease in children, and in the U.S. swine industry at pharmacological levels to promote growth and fecal consistency, but underlying mechanisms explaining these beneficial effects remain unknown. Moreover, adding supplemental phytase improves Zn bioavailability. Thus, we hypothesized that bene ...[more]
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Project description:Zinc (Zn) is an essential trace element for all life forms. Zn supplementation has been used to treat diarrheal disease in children, and in the U.S. swine industry at pharmacological levels to promote growth and fecal consistency, but underlying mechanisms explaining these beneficial effects remain unknown. Thus, we hypothesized that the benefits of pharmacological Zn supplementation were a result of changes in gene expression. For this study, liver RNA from newly weaned pigs fed dietary Zn as Zn oxide for 14 days at either adequate (150 Zn/kg) or pharmacological (2000 mg Zn/kg) levels was evaluated using a 70-mer oligonucleotide microarray. Interrogation of this microarray revealed 658 annotated transcripts (FDR ≤ 0.05) affected by pharmacological Zn supplementation. Relative real-time RT-PCR was used to confirm differential expression of two genes. Results suggest that feeding pharmacological Zn (2000 mg Zn/kg) affects genes involved in reducing oxidative stress and in amino acid metabolism, which are essential for cell detoxification and proper cell function.
2008-10-02 | GSE11972 | GEO
Project description:Zinc (Zn) is an essential trace element for all life forms. Zn supplementation has been used to treat diarrheal disease in children, and in the U.S. swine industry at pharmacological levels to promote growth and fecal consistency, but underlying mechanisms explaining these beneficial effects remain unknown. Thus, we hypothesized that the benefits of pharmacological Zn supplementation were a result of changes in gene expression. For this study, liver RNA from newly weaned pigs fed dietary Zn as Zn oxide for 14 days at either adequate (150 Zn/kg) or pharmacological (2000 mg Zn/kg) levels was evaluated using a 70-mer oligonucleotide microarray. Interrogation of this microarray revealed 658 annotated transcripts (FDR ≤ 0.05) affected by pharmacological Zn supplementation. Relative real-time RT-PCR was used to confirm differential expression of two genes. Results suggest that feeding pharmacological Zn (2000 mg Zn/kg) affects genes involved in reducing oxidative stress and in amino acid metabolism, which are essential for cell detoxification and proper cell function. Oligonucleotide microarrays used for this study consisted of 13,297 70-mer oligos (Pig Array-Ready Oligo Set v. 1.0 and Pig Oligo Extension Set v. 1.0, Qiagen, Inc., Valencia, CA) each spotted once on a single slide. Slides were printed at the Michigan State University Research Technology Support Facility. Oligonucleotides spotted in multiple locations for use as potential controls included 76 Arabidopsis thaliana gene spots, 17 beta tubulin spots, 17 glyceraldehyde-3-phosphate dehydrogenase spots, 85 heat shock protein gene spots, 69 ribosomal protein gene spots, 112 randomly generated negative control spots and 470 blanks. The microarray was screened with the Zn150 and Zn2000 samples, and four microarray slides were screened. Zn150 samples were randomly paired with Zn2000 samples. Two samples from each treatment were labeled with Cy3 and the other two were labeled with Cy5.
2008-10-02 | E-GEOD-11972 | biostudies-arrayexpress
Project description:Phytase supplementation in broiler diets is a common practice to improve phosphorus (P) availability and to reduce P loss by excretion. An enhanced P availability, and its concomitant supplementation with calcium (Ca), can affect the structure of the microbial community in the digestive tract of broiler chickens. Here, we aim to distinguish the effects of mineral P, Ca, and phytase on the composition of microbial communities present in the content and the mucosa layer of the gastrointestinal tract (GIT) of broiler chickens. Significant differences were observed between digesta and mucosa samples for the GIT sections studied (p = 0.001). The analyses of 56 individual birds showed a high microbial composition variability within the replicates of the same diet. The average similarity within replicates of digesta and mucosa samples across all diets ranged from 29 to 82% in crop, 19-49% in ileum, and 17-39% in caeca. Broilers fed with a diet only supplemented with Ca had the lowest body weight gain and feed conversion values while diets supplemented with P showed the best performance results. An effect of each diet on crop mucosa samples was observed, however, similar results were not obtained from digesta samples. Microbial communities colonizing the ileum mucosa samples were affected by P supplementation. Caeca-derived samples showed the highest microbial diversity when compared to the other GIT sections and the most prominent phylotypes were related to genus Faecalibacterium and Pseudoflavonifractor, known for their influence on gut health and as butyrate producers. Lower microbial diversity in crop digesta was linked to lower growth performance of birds fed with a diet only supplemented with Ca. Each diet affected microbial communities within individual sections, however, no diet showed a comprehensive effect across all GIT sections, which can primarily be attributed to the great variability among replicates. The substantial community differences between digesta and mucosa derived samples indicate that both habitats have to be considered when the influence of diet on the gut microbiota, broiler growth performance, and animal health is investigated.
| S-EPMC5165256 | biostudies-literature
Project description:The objective of this study was to evaluate the effects of dietary xylanase (X) and a carbohydrase enzyme blend (EB: cellulase, ?-glucanase, and xylanase) on nutrient digestibility, intestinal barrier integrity, inflammatory status, and growth performance in weaned piglets fed higher fiber diets. A total of 460 pigs (6.43 ± 0.06 kg BW; F25 × 6.0 Genetiporc) were blocked by initial BW and pens (n = 12 per treatment) were randomly assigned to 1 of 4 dietary treatments. The diets included a higher fiber unsupplemented control diet (CON) and the CON supplemented with 0.01% X, 0.01% EB, or both enzymes, arranged in a 2 × 2 factorial. The diets were based on corn, soybean meal, corn distillers dried grains with solubles (DDGS), and wheat middlings. Pigs had 7 d to adapt to the environment and consumed the same commercial diet. Pigs were fed the experimental diets for 28 d with free access to feed and water. Body weight and feed disappearance were recorded weekly. One pig with BW closest to the pen average from each pen was selected and moved to metabolism crates on day 16 and intragastric gavaged a solution of lactulose and mannitol on day 22 followed by 12-h urine collection. Feces were collected from day 23 to 25. Intestinal tissues and mucosal scrapings were collected on day 28. Data were analyzed using PROC MIXED of SAS (9.4). Xylanase, EB, and their interaction were fixed effects and block was a random effect. The EB, but not X, increased pig BW and improved ADG over 28 d (P < 0.05). Neither carbohydrase impacted ADFI, G:F, or apparent total tract digestibility (ATTD) of DM, GE, or CP. The EB improved ATTD of ADF (32.45 vs. 26.57%; P < 0.01), but had no effect on NDF. Unexpectedly, X reduced ATTD of NDF and ADF (P < 0.01). The EB reduced urinary lactulose:mannitol and increased ileal claudin-3 mRNA abundance (P < 0.05), indicating improved small intestinal barrier integrity. There was a X × EB interaction on ileal secretory immunoglobulin A (sIgA) concentration (P < 0.05); in the absence of X, EB decreased sIgA compared to CON, but this effect disappeared in the presence of X. The EB also reduced ileal IL-22 mRNA abundance (P < 0.05), probably indicating decreased immune activation. In conclusion, EB but not X enhanced growth rate of weaned pigs fed higher fiber diets, which may be partly explained by the improved small intestinal barrier integrity and reduced immune activation, rather than improvement in nutrient digestibility.
| S-EPMC6276555 | biostudies-literature
Project description: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.
| S-EPMC9219510 | biostudies-literature
Project description:Pharmacological levels of zinc oxide can promote growth and health of weaning piglets, but the underlying molecular mechanisms are yet not fully understood. The aim of this study was to determine changes in the global hepatic protein expression in response to dietary zinc oxide in weaned piglets. Nine half-sib piglets were allocated to three dietary zinc treatment groups (50, 150, 2500 mg/kg dry matter). After 14 d, pigs were euthanized and liver samples taken. The increase in hepatic zinc concentration following dietary supplementation of zinc was accompanied by up-regulation of metallothionein mRNA and protein expression. Global hepatic protein profiles were obtained by two-dimensional difference gel electrophoresis following matrix-assisted laser desorption ionization/time-of-flight mass spectrometry. A total of 15 proteins were differentially (P<0.05) expressed between groups receiving control (150 mg/kg) or pharmacological levels of zinc (2500 mg/kg) with 7 down- (e.g. arginase1, thiosulfate sulfurtransferase, HSP70) and 8 up-regulated (e.g. apolipoprotein AI, transferrin, C1-tetrahydrofolate synthase) proteins. Additionally, three proteins were differentially expressed with low zinc supply (50 mg/kg Zn) in comparison to the control diet. The identified proteins were mainly associated with functions related to cellular stress, transport, metabolism, and signal transduction. The differential regulation was evaluated at the mRNA level and a subset of three proteins of different functional groups was selected for confirmation by western blotting. The results of this proteomic study suggest that zinc affects important liver functions such as blood protein secretion, protein metabolism, detoxification and redox homeostasis, thus supporting the hypothesis of intermediary effects of pharmacological levels of zinc oxide fed to pigs.
| S-EPMC3839893 | biostudies-literature
Project description:Strategies to reduce economic losses associated with post-weaning diarrhea in pig farming include high-level dietary zinc oxide supplementation. However, excessive usage of zinc oxide in the pig production sector was found to be associated with accumulation of multidrug resistant bacteria in these animals, presenting an environmental burden through contaminated manure. Here we report on zinc tolerance among a random selection of intestinal Escherichia coli comprising of different antibiotic resistance phenotypes and sampling sites isolated during a controlled feeding trial from 16 weaned piglets: In total, 179 isolates from "pigs fed with high zinc concentrations" (high zinc group, [HZG]: n = 99) and a corresponding "control group" ([CG]: n = 80) were investigated with regard to zinc tolerance, antimicrobial- and biocide susceptibilities by determining minimum inhibitory concentrations (MICs). In addition, in silico whole genome screening (WGSc) for antibiotic resistance genes (ARGs) as well as biocide- and heavy metal tolerance genes was performed using an in-house BLAST-based pipeline. Overall, porcine E. coli isolates showed three different ZnCl2 MICs: 128 μg/ml (HZG, 2%; CG, 6%), 256 μg/ml (HZG, 64%; CG, 91%) and 512 μg/ml ZnCl2 (HZG, 34%, CG, 3%), a unimodal distribution most likely reflecting natural differences in zinc tolerance associated with different genetic lineages. However, a selective impact of the zinc-rich supplemented diet seems to be reasonable, since the linear mixed regression model revealed a statistically significant association between "higher" ZnCl2 MICs and isolates representing the HZG as well as "lower ZnCl2 MICs" with isolates of the CG (p = 0.005). None of the zinc chloride MICs was associated with a particular antibiotic-, heavy metal- or biocide- tolerance/resistance phenotype. Isolates expressing the 512 μg/ml MIC were either positive for ARGs conferring resistance to aminoglycosides, tetracycline and sulfamethoxazole-trimethoprim, or harbored no ARGs at all. Moreover, WGSc revealed a ubiquitous presence of zinc homeostasis and - detoxification genes, including zitB, zntA, and pit. In conclusion, we provide evidence that zinc-rich supplementation of pig feed selects for more zinc tolerant E. coli, including isolates harboring ARGs and biocide- and heavy metal tolerance genes - a putative selective advantage considering substances and antibiotics currently used in industrial pork production systems.
| S-EPMC6892955 | biostudies-literature
Project description:Body phosphorus homeostasis network allows laying hens to adapt to wide range of changes in dietary phosphorus levels. Phytase hydrolyzes phytate rendering phosphorus and reduces the laying hens' requirements for inorganic phosphate rock. Here, we demonstrate that there is no need to keep large safety margins in dietary phosphorus when hens are fed with phytase. Hy-Line Brown laying hens (n = 504) were randomly assigned to 7 treatments (6 replicates of 12 birds). A corn-soybean meal-based diet, with no inorganic phosphate rock, was formulated to contain 0.12% nonphytate phosphorus (nPP), 3.8% calcium, and 2,000 FTU/kg phytase. Inorganic phosphate rock (di-calcium phosphate) was supplemented into the basal diet to create 6 other diets containing 0.17, 0.22, 0.27, 0.32, 0.37, and 0.42% nPP. Levels of calcium carbonate and zeolite powder were adjusted to make sure all the 7 experimental diets contained the same nutrition levels (including calcium and phytase) except nPP. The diets were subjected to laying hens from 29 to 40 wk of age. As a result, when supplemented with 2,000 FTU/kg phytase, extra supplementation of inorganic phosphate rock had no effects (P > 0.05) on serum phosphorus levels, serum calcium levels, laying performance (laying rate, egg weight, feed intake, feed-to-egg ratio, and unqualified egg rate), egg quality (shell thickness, shell strength, albumen height, yolk color, and Haugh unit), and tibia quality parameters (breaking strength and ash, calcium, and phosphorus contents). Extra supplementation of inorganic phosphate rock linearly increased (P < 0.01) fecal phosphorus excretion and linearly decreased (P = 0.032) the apparent metabolizability of dietary phosphorus. While serum hormones and intestine gene expressions were varied within treatments, no consistent changes were found. In conclusion, the supplementation of inorganic phosphate rock (provided 0.05-0.30% extra nPP) to phytase-containing basal diets (2,000 FTU/kg; nPP = 0.12%) provided limited benefits to egg production performance in laying hens from 29 to 40 wk of age. Further investigating the body phosphorus homeostasis would help to understand the nutritional and physiological reasonability of formulating low-phosphorus diets in the laying hen industry.
| S-EPMC7597811 | biostudies-literature
Project description:Glyphosate possesses antimicrobial properties, and the present study investigated potential effects of feed glyphosate on piglet gastrointestinal microbial ecology. Weaned piglets were allocated to four diets (glyphosate contents [mg/kg feed]: 0 mg/kg control [CON; i.e., basal diet with no glyphosate added], 20 mg/kg as Glyphomax commercial herbicide [GM20], and 20 mg/kg [IPA20] and 200 mg/kg [IPA200] as glyphosate isopropylamine [IPA] salt). Piglets were sacrificed after 9 and 35 days of treatment, and stomach, small intestine, cecum, and colon digesta were analyzed for glyphosate, aminomethylphosphonic acid (AMPA), organic acids, pH, dry matter content, and microbiota composition. Digesta glyphosate contents reflected dietary levels (on day 35, 0.17, 16.2, 20.5, and 207.5 mg/kg colon digesta, respectively). Overall, we observed no significant glyphosate-associated effects on digesta pH, dry matter content, and-with few exceptions-organic acid levels. On day 9, only minor gut microbiota changes were observed. On day 35, we observed a significant glyphosate-associated decrease in species richness (CON, 462; IPA200, 417) and in the relative abundance of certain Bacteroidetes genera: CF231 (CON, 3.71%; IPA20, 2.33%; IPA200, 2.07%) and g_0.24 (CON, 3.69%; IPA20, 2.07%; IPA200, 1.75%) in cecum. No significant changes were observed at the phylum level. In the colon, we observed a significant glyphosate-associated increase in the relative abundance of Firmicutes (CON, 57.7%; IPA20, 69.4%; IPA200, 66.1%) and a decrease in Bacteroidetes (CON, 32.6%; IPA20, 23.5%). Significant changes were only observed for few genera, e.g., g_0.24 (CON, 7.12%; IPA20, 4.59%; IPA200, 4.00%). In conclusion, exposing weaned piglets to glyphosate-amended feed did not affect gastrointestinal microbial ecology to a degree that was considered actual dysbiosis, e.g., no potential pathogen bloom was observed. IMPORTANCE Glyphosate residues can be found in feed made from genetically modified glyphosate-resistant crops treated with glyphosate or from conventional crops, desiccated with glyphosate before harvest. If these residues affect the gut microbiota to an extent that is unfavorable to livestock health and productivity, the widespread use of glyphosate on feed crops may need to be reconsidered. Few in vivo studies have been conducted to investigate potential impact of glyphosate on the gut microbial ecology and derived health issues of animals, in particular livestock, when exposed to dietary glyphosate residues. The aim of the present study was therefore to investigate potential effects on the gastrointestinal microbial ecology of newly weaned piglets fed glyphosate-amended diets. Piglets did not develop actual gut dysbiosis when fed diets, containing a commercial herbicide formulation or a glyphosate salt at the maximum residue level, defined by the European Union for common feed crops, or at a 10-fold-higher level.
| S-EPMC10433988 | biostudies-literature
Project description:Zinc oxide (ZnO) is commonly fed to pigs at pharmacologic concentrations (2,000-3,000 ppm) for the first 3 wk post-weaning to increase growth and reduce enteric bacterial disease. The safety of this high-dose treatment is assumed based upon lower bioavailability of ZnO compared to other common forms of Zn in feed; however, limited data are available regarding the specific serum and tissue concentrations of Zn expected in animals experiencing overload following feeding of excessive ZnO. Fifty-five 3-wk-old pigs were divided into 5 groups receiving various concentrations of ZnO (0-6,000 ppm) for 3 wk. Pigs receiving 6,000 ppm ZnO had higher mean pancreatic Zn concentrations (p < 0.001) compared to other treatments, and higher pancreatic Zn concentrations were associated with pancreatic acinar cell apoptosis (p < 0.0001). Hepatic Zn concentrations were highest for pigs receiving 6,000 ppm ZnO (mean ± SEM; 729 ± 264 ppm) and significantly higher than all other groups (p < 0.0001), with controls having concentrations <60 ppm. Similarly, serum Zn was highest in pigs receiving 6,000 ppm ZnO (4.81 ± 2.31 ppm) and significantly higher than all groups (controls, <1 ppm). Additionally, as pigs became overloaded with Zn, there were significant reductions in serum Cu and both serum and hepatic Se. Hepatic and serum Zn concentrations >500 ppm and >2 ppm, respectively, are indicative of Zn overload, and dietary trace mineral analysis is warranted if expected inclusion rates are ?3,000 ppm ZnO.
| S-EPMC6857036 | biostudies-literature