Embryo injected with Ochratoxin A induced jejunum injury in ducklings by activating the TLR4 signaling pathway: Involvement of intestinal microbiota.

Ochratoxin A (OTA) is a common mycotoxin that causes intestinal injury in humans and various animal species. OTA may lead to intestinal injury in offspring due to the maternal effect. The aim of this study was to investigate the mechanism of embryo injected with OTA induced jejunum injury in ducklings. The results showed that OTA disrupted the jejunum tight junctions in hatching ducklings, and promoted the secretion of inflammatory cytokines. And this inflammatory response was caused by the activation of the TLR4 signaling pathway. Moreover, embryo injected with OTA could cause damage to the intestinal barrier in 21-day-old ducks, characterized by shortened villi, crypt hyperplasia, disrupted intestinal tight junctions, increased level of LPS in the jejunum, activation of the TLR4 signaling pathway, and increased levels of pro-inflammatory cytokines. Meanwhile, OTA induced oxidative stress in the jejunum. And dysbiosis of gut microbiota was mainly characterized by an increased the relative abundance of Bacteroides, Megamonas, Fournierella, and decreased the relative abundance of Alistipes and Weissella. Interestingly, embryo injected with OTA did not induce these changes in the jejunum of antibiotics-treated 21-day-old ducks. In conclusion, embryo injected with OTA induced jejunum injury in ducklings by activating the TLR4 signaling pathway, which involvement of intestinal microbiota.

Ochratoxin A induces abnormal tryptophan metabolism in the intestine and liver to activate AMPK signaling pathway.

BACKGROUND: Ochratoxin A (OTA) is a mycotoxin widely present in raw food and feed materials and is mainly produced by Aspergillus ochraceus and Penicillium verrucosum. Our previous study showed that OTA principally induces liver inflammation by causing intestinal flora disorder, especially Bacteroides plebeius (B. plebeius) overgrowth. However, whether OTA or B. plebeius alteration leads to abnormal tryptophan-related metabolism in the intestine and liver is largely unknown. This study aimed to elucidate the metabolic changes in the intestine and liver induced by OTA and the tryptophan-related metabolic pathway in the liver. MATERIALS AND METHODS: A total of 30 healthy 1-day-old male Cherry Valley ducks were randomly divided into 2 groups. The control group was given 0.1 mol/L NaHCO3 solution, and the OTA group was given 235 µg/kg body weight OTA for 14 consecutive days. Tryptophan metabolites were determined by intestinal chyme metabolomics and liver tryptophan-targeted metabolomics. AMPK-related signaling pathway factors were analyzed by Western blotting and mRNA expression. RESULTS: Metabolomic analysis of the intestinal chyme showed that OTA treatment resulted in a decrease in intestinal nicotinuric acid levels, the downstream product of tryptophan metabolism, which were significantly negatively correlated with B. plebeius abundance. In contrast, OTA induced a significant increase in indole-3-acetamide levels, which were positively correlated with B. plebeius abundance. Simultaneously, OTA decreased the levels of ATP, NAD+ and dipeptidase in the liver. Liver tryptophan metabolomics analysis showed that OTA inhibited the kynurenine metabolic pathway and reduced the levels of kynurenine, anthranilic acid and nicotinic acid. Moreover, OTA increased the phosphorylation of AMPK protein and decreased the phosphorylation of mTOR protein. CONCLUSION: OTA decreased the level of nicotinuric acid in the intestinal tract, which was negatively correlated with B. plebeius abundance. The abnormal metabolism of tryptophan led to a deficiency of NAD+ and ATP in the liver, which in turn activated the AMPK signaling pathway. Our results provide new insights into the toxic mechanism of OTA, and tryptophan metabolism might be a target for prevention and treatment.

Effects of embryo injected with ochratoxins A on hatching quality and jejunum antioxidant capacity of ducks at hatching.

Numerous studies have shown that ochratoxins A (OTA) exerts diverse toxicological effects, namely, hepatotoxicity, nephrotoxicity, genotoxicity, enterotoxicity, and immunotoxicity. The main objective of this study was to investigate the influence of embryonic exposure to OTA by different injection times and OTA doses on hatching quality and jejunal antioxidant capacity of ducks at hatching. In total, 480 fertilized eggs were weighed and randomly assigned into a 4 × 4 factorial design including four OTA doses (0, 2, 4, and 8 ng/g egg) on 8, 13, 18, and 23 of embryonic development (E8, E13, E18, and E23). Each treatment included 6 repeats with 5 eggs per repeat. The results showed that the injection time affected the hatching weight (P < 0.0001). The relative length of the jejunum and ileum on E18 and E23 was lower than on E8 and E13 (P < 0.05). Injection time, doses, and their interaction had no effect on jejunum morphology, namely, villous height (Vh), crypt depth (Cd), and villous height/crypt depth ratio Vh/Cd (P > 0.05). The injection time affected the activities of Superoxide dismutase (SOD) (P < 0.0001), total antioxidant capacity (T-AOC) (P < 0.05) and the malondialdehyde (MDA) content (P < 0.0001). The activity of SOD and T-AOC activities in the jejunum of ducklings injected with OTA at the E8 and E13 was lower than that injected at the E18 (P < 0.05). The highest MDA content was observed in ducklings injected with OTA at the E13 (P < 0.05). The injection time (P < 0.0001), OTA doses and their interaction affected the contents of IL-1ß (P < 0.05), which significantly increased especially on E13. In conclusion, the embryo injected with ochratoxins A affected the hatching weight, the relative length of jejunum and ileum, decreased the antioxidant capacity and increased the content of proinflammatory cytokine IL-1ß of the jejunum.

Ochratoxin A: its impact on poultry gut health and microbiota, an overview.

Ochratoxin A (OTA) is a widespread mycotoxin, that has strong thermal stability, and is difficult to remove from feed. OTA is nephrotoxic, hepatotoxic, teratogenic, immunotoxic, and enterotoxic to several species of animals. The gut is the first defense barrier against various types of mycotoxins present in feed that enter the body, and it is closely connected to other tissues through enterohepatic circulation. Compared with mammals, poultry is more sensitive to OTA and has a lower absorption rate. Therefore, the gut is an important target tissue for OTA in poultry. This review comprehensively discusses the role of OTA in gut health and the gut microbiota of poultry, focusing on the effect of OTA on digestive and absorptive processes, intestinal barrier integrity, intestinal histomorphology, gut immunity, and gut microbiota. According to the studies described to date, OTA can affect gut dysbiosis, including increasing gut permeability, immunity, and bacterial translocation, and can eventually lead to gut and other organ injury. Although there are many studies investigating the effects of OTA on the gut health of poultry, further studies are needed to better characterize the underlying mechanisms of action and develop preventative or therapeutic interventions for mycotoxicosis in poultry.

Melatonin alleviates Ochratoxin A-induced liver inflammation involved intestinal microbiota homeostasis and microbiota-independent manner.

Melatonin (MEL) shows an anti-inflammatory effect and regulates intestinal microbiota communities in animals and humans; Ochratoxin A (OTA) induces liver inflammation through intestinal microbiota. However, it remains to know whether MEL alleviates the liver inflammation induced by OTA. In this study, MEL reversed various adverse effects induced by OTA. MEL recovered the swarming and motility of intestinal microbiota, decreased the accumulation of lipopolysaccharide (LPS), enhanced the tight junction proteins of jejunum and cecum segments; ultimately alleviated OTA-induced liver inflammation in ducks. However, it is worth noting that MEL still had positive effects on the OTA-exposed ducks after antibiotic treatment. These results suggest that both the maintenance of intestinal microbiota homeostasis and intestinal microbiota-independent manner involved the MEL anti-inflammatory function in OTA-induced liver inflammation. MEL represent a promising protective approach for OTA, even other mycotoxins.

Effects of Selenium-Enriched Yeast on Performance, Egg Quality, Antioxidant Balance, and Egg Selenium Content in Laying Ducks.

This study investigated the effects of dietary selenium-enriched yeast (Se yeast) supplementation on the laying performance, egg quality, plasma antioxidant balance, and egg selenium (Se) content in laying Longyan ducks. A total of 480 32-week-old ducks were randomly divided into four dietary treatments, each consisting of six replicates of 20 ducks. The dietary treatments were a control basal diet and basal diets with supplementation of 0.05, 0.15, and 0.25 mg Se/kg via Se yeast. The analyzed Se contents of the four diets were 0.15, 0.21, 0.36, and 0.43 mg Se/kg, respectively. Dietary Se yeast supplementation had no apparent effects on laying performance and egg quality (p > 0.05), but it improved the antioxidant balance of ducks, as inferred by greater glutathione peroxidase and catalase activities, and decreased the malondialdehyde content in plasma of ducks (p < 0.05). It was suggested that the Se content in the basal diet containing 0.15 mg/kg of Se requirement is adequate for productive performance, but not for the antioxidant balance of laying ducks. Besides that, the Se contents in the yolk, albumen, and whole egg increased linearly as the Se supplementation levels increased. With more feeding days, the Se contents in the yolk and whole egg from non-Se-yeast-supplemented ducks increased linearly (p < 0.05), while those from Se-yeast-supplemented ducks showed a quadratic relationship (p < 0.05). In conclusion, the Se content of the basal diet at 0.15 mg/kg was adequate for laying performance and egg quality traits in laying ducks. Dietary Se yeast supplementation is beneficial to improve the antioxidant balance of laying ducks and increase the Se deposition in eggs for producing Se-enriched eggs. Based on the quadratic model or the quadratic broken-line model analyses, supplemental 0.19 mg Se/kg via Se yeast, with a total equivalent of 0.34 mg Se/kg in the diet, could provide the optimum antioxidant balance in laying ducks. Dietary supplementation of 0.25 mg Se/kg via Se yeast, with a total equivalent of 0.40 mg Se/kg in the diet, could lead to achieving the desired Se content in the whole egg.

Effect of dietary Moringa stem meal level on growth performance, slaughter performance and serum biochemical parameters in geese.

Moringa stem meal (MSM) with a high level of crude fibre (CF) might be developed and utilized in herbivorous geese as an unconventional feedstuff. The aim of this study was to investigate the effect of the MSM level in the diet on the growth performance, slaughter performance, breast meat quality and serum biochemical parameters in geese from 22 to 70 days of age. A one-factor completely randomized design was adopted in our study. A total of one thousand eight 21-day-old geese were randomly divided into six groups, with six replicates per group and 28 birds per replicate. The geese were fed diets containing MSM levels of 0, 20, 40, 60, 80 or 100 g/kg during day 22-70. The dietary MSM level had no effect (p > .05) on the final body weight (BW), average daily gain (ADG) or average daily feed intake (ADFI). The feed/gain ratio (F/G) increased linearly (p < .001) as the dietary MSM level increased. No differences (p > .05) were observed in the slaughter performance, meat quality and the relative organ weight (except for thymus) of the geese (p > .05). The relative weight of the thymus in the geese fed diets with supplementation of MSM was higher than that in the non-supplemented MSM control group (p < .05). In addition, 100 g MSM/kg of diet decreased the serum glucose (GLU) level (p < .05) and increased the alanine transaminase (ALT) enzyme activity (p = .03). Dietary MSM levels of no more than 60 g/kg had no effects on the growth performance and slaughter performance, whereas diets with 100 g MSM/kg increased the F/G and serum ALT enzyme activity, as well as decreasing the serum GLU level. Therefore, MSM provided at a reasonable level could be developed as an unconventional feedstuff for geese at the finisher period.

Ochratoxin A induces liver inflammation: involvement of intestinal microbiota.

BACKGROUND: Ochratoxin A (OTA) is a widespread mycotoxin and induces liver inflammation to human and various species of animals. The intestinal microbiota has critical importance in liver inflammation; however, it remains to know whether intestinal microbiota mediates the liver inflammation induced by OTA. Here, we treated ducklings with oral gavage of OTA (235 µg/kg body weight) for 2 weeks. Then, the microbiota in the cecum and liver were analyzed with 16S rRNA sequencing, and the inflammation in the liver was analyzed. To explore the role of intestinal microbiota in OTA-induced liver inflammation, intestinal microbiota was cleared with antibiotics and fecal microbiota transplantation was conducted. RESULTS: Here, we find that OTA treatment in ducks altered the intestinal microbiota composition and structure [e.g., increasing the relative abundance of lipopolysaccharides (LPS)-producing Bacteroides], and induced the accumulation of LPS and inflammation in the liver. Intriguingly, in antibiotic-treated ducks, OTA failed to induce these alterations in the liver. Notably, with the fecal microbiota transplantation (FMT) program, in which ducks were colonized with intestinal microbiota from control or OTA-treated ducks, we elucidated the involvement of intestinal microbiota, especially Bacteroides, in liver inflammation induced by OTA. CONCLUSIONS: These results highlight the role of gut microbiota in OTA-induced liver inflammation and open a new window for novel preventative or therapeutic intervention for mycotoxicosis.

Effects of Dietary n-6:n-3 PUFA Ratios on Lipid Levels and Fatty Acid Profile of Cherry Valley Ducks at 15-42 Days of Age.

The objective of this study was to investigate the effects of dietary n-6:n-3 PUFA ratio on growth performance, serum and tissue lipid levels, fatty acid profile, and hepatic expression of fatty acid synthesis genes in ducks. A total of 3168 15-day old ducks were fed different n-6:n-3 PUFA ratios: 13:1 (control), 10:1, 8:1, 6:1, 4:1, and 2:1. The feeding trial lasted 4 weeks. Our results revealed that dietary n-6:n-3 PUFA ratios had no effects on growth performance. The 2:1 group had the highest serum triglyceride levels. Serum total cholesterol and HDL levels were higher in the 13:1 and 8:1 groups than in the 6:1 and 2:1 groups. The concentration of C18:3n-3 in serum and tissues (liver and muscle) increased with decreasing dietary n-6:n-3 PUFA ratios. The hepatic expression of FADS2, ELOVL5, FADS1, and ELOVL2 increased on a quadratic function with decreasing dietary n-6:n-3 PUFA ratios. These results demonstrate that lower dietary n-6:n-3 PUFA ratios had strong effects on the fatty acid profile of edible parts and the deposition of n-3 PUFAs in adipose tissue of ducks.

Effects of dietary protein and energy levels on digestive enzyme activities and electrolyte composition in the small intestinal fluid of geese.

The present study was conducted to evaluate the effects of dietary protein and energy levels on digestive enzymes and electrolyte composition in jejunum of geese. A 3×3 factorial and completely randomized design was adopted with three protein levels and three energy levels. The experiment included four replicates for each treatment, and three geese for each replicate. Isovolumetric supernate from centrifugal jejuna fluid were mixed in each replicate. Activities of digestive enzymes and ions were analyzed. The results showed trypsin and chymotrypsin activities were significantly increased with increasing of dietary protein and energy levels (P<0.05). The concentrations of Ca2+ and pH value were significantly decreased by increased dietary protein and energy levels. However, no significant differences were found for the activities of amylase and cellulase, as well as the concentration of Na+ among groups with different protein and energy levels. In conclusion, digesta enzymes and electrolytes in the small intestine adapted to the protein and energy levels. The activities of protease, rather than amylase and cellulase were induced with increasing of protein and energy levels. The imbalance of positive and negative ions was possibly adjusted by the fluctuant concentrations of K+ , Cl- and Ca2+ for maintaining normal physiological function.