Dietary deoxynivalenol (DON) impairs the intestinal functions and performance in broiler chickens, whereas little is known about the effect of DON on the gastrointestinal microbiota. This study evaluated the impact of graded levels of dietary DON contamination on the cecal bacterial microbiota, their predicted metabolic abilities and short-chain fatty acid (SCFA) profiles in chickens. In using a single oral lipopolysaccharide (LPS) challenge we further assessed whether an additional intestinal stressor would potentiate DON-related effects on the cecal microbiota. Eighty 1-day-old chicks were fed diets with increasing DON concentrations (0, 2.5, 5, and 10 mg DON per kg diet) for 5 weeks and were sampled after half of the chickens received an oral LPS challenge (1 mg LPS/kg bodyweight) 1 day before sampling. The bacterial composition was investigated by Illumina MiSeq sequencing of the V3–5 region of the 16S rRNA gene. DON-feeding decreased (p < 0.05) the cecal species richness (Chao1) and evenness (Shannon) compared to the non-contaminated diet. The phyla Firmicutes and Proteobacteria tended to linearly increase and decrease with increasing DON-concentrations, respectively. Within the Firmicutes, DON decreased the relative abundance of Oscillospira, Clostridiaceae genus, Clostridium, and Ruminococcaceae genus 2 (p < 0.05), whereas it increased Clostridiales genus 2 (p < 0.05). Moreover, increasing DON levels linearly decreased a high-abundance Enterobacteriaceae genus and an Escherichia/Shigella-OTU (p < 0.05). Changes in the bacterial composition and their imputed metagenomic capabilities may be explained by DON-related changes in host physiology and cecal nutrient availability. The oral LPS challenge only decreased the abundance of an unassigned Clostridiales genus 2 (p = 0.03). Increasing dietary concentrations of DON quadratically increased the cecal total SCFA and butyrate concentration (p < 0.05), whereas a DON × LPS interaction indicated that LPS mainly increased cecal total SCFA, butyrate, and acetate concentrations in chickens fed the diets that were not contaminated with DON. The present findings showed that even the lowest level of dietary DON contamination had modulatory effects on chicken's cecal bacterial microbiota composition and diversity, whereas the additional oral challenge with LPS did not potentiate DON effects on the cecal bacterial composition.
The aim of the study was to evaluate the effects of low to moderate oral exposure to the Fusarium toxin deoxynivalenol (DON; derived from culture material) on performance, water intake, and carcass parameters of broilers during early and late developmental phases. A total of 160 Ross 308 broilers were randomly allocated to four different feeding groups (n = 40/group) including 0 (control), 2.5, 5, and 10 mg DON/kg wheat-soybean meal-based feed. Three consecutive replicates of the experiment were performed. Half of the broilers were slaughtered in week 3 of the trial whereas the other half were slaughtered in week 5. Dry matter intake (DMI) and water intake (WI) were recorded on a daily basis and the body weight (BW) and BW gain (BWG) were determined weekly. The following carcass traits were recorded and calculated in absolute and relative data: dressed carcass weight, breast muscle weight, leg weight, and liver weight. Data showed that BW (P < 0.001), BWG (P = 0.005), and DMI (P < 0.001) were reduced by DON-feeding during the entire feeding period. The ratio of DMI to body weight gain (DMI/BWG) was not affected by the treatment. However, the ratio of water to DMI (WI/DMI) increased in DON-treated birds (P = 0.021). Contrast analysis showed that DON tendentially reduced slaughter weight (P = 0.082) and decreased leg yield (P = 0.037) in DON-fed chickens in week 5 of the experiment. Liver organ weight decreased in the 3-week-old DON-fed broilers compared to that in the control-fed birds (P = 0.037). In conclusion, the study suggests that DMI and BW were negatively affected under the experimental conditions at DON levels lower than the current guidance value in the European Union of 5 mg/kg feed. The study also indicates that broilers fed on low to moderate level DON-contaminated diets showed increased WI/DMI ratio which might have negative influence on wet litter syndrome.
Dietary deoxynivalenol (DON) impairs the intestinal immune system and digestive functions of broiler chickens. However, little is known whether increasing doses of DON similarly affect the intestinal functions in different segments of the small intestine in chickens and whether a second oral challenge may potentiate those effects. The present objective was to investigate the effect of increasing dietary DON concentrations on the relative expression of genes for tight junction proteins, mucins, toll-like receptors (TLR), and cytokines in duodenum and jejunum, jejunal mucosal permeability, as well as on α-1acid glycoprotein and IgA in serum with or without an additional oral lipopolysaccharide (LPS) challenge. Eighty 1-d-old chickens were fed diets with increasing DON concentrations (0, 2.5, 5, and 10 mg DON/kg diet) for 5 wk. One day before sampling, half of the chickens received an oral challenge with 1-mg Escherichia coli O55:B5 LPS/ kg BW. Ussing chambers were used to measure the jejunal permeability in birds receiving 10-mg DON/kg feed with or without LPS challenge and 0-mg DON/kg feed without LPS. Increasing DON concentrations of up to 5-mg DON/kg increased (P < 0.05) the duodenal expression of TLR2, IL6, and Claudin 1 (CLDN1) by up to 84%, 88%, and 48%, respectively, compared with the noncontaminated diet. Likewise, jejunal CLDN1 expression increased up to 23% in the chickens fed DON concentrations of up to 5-mg DON/kg diet (P < 0.05). Moreover, increasing DON concentrations linearly and quadratically decreased (P < 0.05) the jejunal expression of TLR2 and transforming growth factor-β 1, respectively. The additional LPS challenge increased (P < 0.040) duodenal occludin expression by 10% as well as the jejunal tissue conductance in chickens of the 10 DON group (P = 0.050). In conclusion, dietary DON differently affected the duodenal and jejunal expression of genes for tight-junction proteins and proinflammatory signaling pathways. The additional LPS challenge did not potentiate the DON effect but it seemed to induce a certain up-regulation of the proinflammatory response in the duodenum and enhanced the mucosal permeability in the jejunum.
Exposure to mycotoxin-contaminated feeds represents a serious health risk. This has necessitated the need for the establishment of practical methods for mycotoxin decontamination. This study investigated the effects of citric acid (CA) and lactic acid (LA) on common trichothecene mycotoxins in feeds contaminated with Fusarium mycotoxins. Contaminated feed samples were processed either with 5% CA or 5% LA solutions in a ratio of 1:1.2 (w/v) for 5, 24, or 48 h, and analyzed for multiple mycotoxin metabolites using a liquid chromatography–tandem mass spectrometric method. The analyses showed that treating the feed with CA and LA lowered the concentration of deoxynivalenol (DON), whereby 5% LA lowered the original DON concentration in the contaminated feed samples by half, irrespective of the processing time. Similar lowering effects were observed for the concentrations of 15Ac-DON, 5-hydroxyculmorin, and sambucinol. The concentration of nivalenol was only lowered by the LA treatment. In contrast, CA and LA treatments showed no or only small effects on the concentration of several mycotoxins and their derivatives, including zearalenone, fumonisins, and culmorin. In conclusion, the present results indicate that the use of 5% solutions of LA and CA might reduce the concentration of common trichothecene mycotoxins, especially DON and its derivate 15Ac-DON. However, further research is required to determine the effect on overall toxicity and to identify the underlying mechanisms.
Mycotoxin deactivators are a widely used strategy to abrogate negative effects of mycotoxin-contaminated feed. It has not been adequately evaluated whether these deactivators may detoxify bacterial toxins in the intestinal lumen and subsequently lower the inflammatory response in chickens. The present objective was to study the effect of a multicomponent mycotoxin deactivator (B), containing a bentonite and a bacterial strain capable to enzymatically biotransform trichothecenes especially deoxynivalenol (DON), when supplemented to a DON-contaminated feed in combination with an oral lipopolysaccharide challenge on visceral organ size, expression of innate immune genes and mucosal permeability in the small intestine as well as on the cecal bacterial composition and metabolites in broiler chickens. Eighty 1-d-old male chickens were randomly allotted to four treatment groups in two replicate batches (n = 10/treatment/replicate): 1) basal diet without DON (CON), 2) CON diet supplemented with B (2.5 mg B/kg feed) (CON-B), 3) CON diet contaminated with 10 mg DON/kg feed (DON), and 4) DON diet supplemented with 2.5 mg B/kg feed (DON-B). In half of the chickens per treatment, effects were assessed under nonchallenge conditions, whereas in the other half of birds, to increase their intestinal bacterial toxin load, effects were tested after an oral challenge with 1 mg LPS/kg BW from Escherichia coli O55:B5 on the day before sampling. DON reduced (P < 0.05) the weight of bursa fabricii and thymus. DON increased the expression level of intestinal alkaline phosphatase at the duodenal mucosa (P = 0.027) but did not modify jejunal gene expression and mucosal permeability. The LPS challenge decreased the jejunal MUC2 expression but increased ZO1 and IL6 expression compared to the unchallenged animals (P < 0.05). DON × B interactions indicated lower expression of IL10 in duodenum and NFKB in jejunum with the B diet but higher expression with the DON-B diet (P = 0.050). Furthermore, the B lowered jejunal expression of NFKB and IL6 but only in LPS-challenged chickens (P < 0.05). Alterations in the cecal microbiota composition and VFA profile were likely associated with alterations in host physiology in the small intestine caused by DON, B, and LPS. According to the present data, B appeared to have potential to detoxify antigens other than DON in the intestinal lumen of chickens, whereby the toxin load may limit the efficacy of B to modify the intestinal and systemic response as indicated by interactions of DON, B, and LPS.
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