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Abstract
Plant materials can be contaminated with Fusarium mycotoxins and their derivatives, whose toxic effects on humans and animals may remain subclinical. Zearalenone (ZEN), a low-molecular-weight compound, is produced by molds in crop plants as a secondary metabolite. The objective of this study will be to analyze the in vivo correlations between very low monotonic doses of ZEN (5, 10, and 15 μg ZEN/kg body weight—BW for 42 days) and the carryover of this mycotoxin and its selected metabolites from the intestinal contents to the intestinal walls, the mRNA expression of estrogen receptor alfa (ERα) and estrogen receptor beta (ERβ) genes, and the mRNA expression of genes modulating selected colon enzymes (CYP1A1 and GSTP1) in the intestinal mucosa of pre-pubertal gilts. An in vivo experiment will be performed on 60 clinically healthy animals with initial BW of 14.5 ± 2 kg. The gilts will be randomly divided into a control group (group C, n = 15) and three experimental groups (group ZEN5, group ZEN10, and group ZEN15; n = 15). Group ZEN5 will be administered per os 5 μg ZEN/kg BW (MABEL), group ZEN10—10 μg ZEN/kg BW (NOAEL), and group ZEN15—15 µg ZEN/kg BW (low LOAEL). In each group, five animals will be euthanized on analytical dates 1 (exposure day 7), 2 (exposure day 21), and 3 (exposure day 42). Samples for in vitro analyses will be collected from an intestinal segment resected from the following regions: the third (horizontal) part of the duodenum, jejunum, ileum, cecum, ascending colon, transverse colon, and descending colon. The experimental material will be collected under special conditions, and it will be transported to specialist laboratories where samples will be obtained for further analyses.
Highlights
A high percentage of plant-based raw materials used in feed production [1] may be contaminated with mycotoxins, increasing the risk of poisoning in humans [2] and farm animals, pigs [3]
The study will expand our understanding of the mechanisms underlying the expression of ERα and Erβ, which participate in both stages of mycotoxin biotransformation
The Cq values obtained for each dilution series will be plotted against the log copy number, and will be used to extrapolate unknown samples to copy numbers. mRNA copy numbers of the samples collected from all experimental groups in each exposure period will be divided by the averaged numbers from the control group, determined at the beginning of the experiment, to obtain relative expression values, which will be presented as the expression ratio (R)
Summary
A high percentage of plant-based raw materials used in feed production [1] may be contaminated with mycotoxins (undesirable substances), increasing the risk of poisoning in humans [2] and farm animals, pigs [3]. In other studies investigating the effects of ZEN biotransformation in peripheral blood, animals were exposed to much higher doses of the mycotoxin [21,50,51] These observations could suggest that in line with the hormesis paradigm [8,21], the exposure to very low doses of ZEN affects the synthesis and secretion of sex steroid hormones [18,39]. The interactions between endogenous and exogenous (environmental) steroids could be influenced by other endogenous factors, such as the accumulation of ZEN and its metabolites in intestinal tissues in the initial stages of biotransformation, the resulting expression of ERs and selected intestinal enzymes that participate in detoxification, which is accompanied by specific (but not clearly determined) accumulation of ZEN and its metabolites in intestinal tissues Undesirable substances such as ZEN are metabolized inside cells by two classes of enzymes. The study will expand our understanding of the mechanisms underlying the expression of ERα and Erβ, which participate in both stages of mycotoxin (undesirable substance) biotransformation
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