RESUMO
Lead (Pb) is a hazardous pollutant in water environments that can cause significant damage to aquatic animals and humans. In this study, crucian carp (Carassius auratus) were exposed to waterborne Pb for 96 h; then, histopathological analysis, quantitative qPCR analysis, and 16S high-throughput sequencing were performed to explore the effects of Pb on intestinal bioaccumulation, structural damage, oxidative stress, immune response, and microbiota imbalance of C. auratus. After Pb exposure, the intestinal morphology was obviously damaged, including significantly increasing the thickness of the intestinal wall and the number of goblet cells and reducing the depth of intestinal crypts. Pb exposure reduced the mRNA expressions of Claudin-7 and villin-1 while significantly elevated the level of GST, GSH, CAT, IL-8, IL-10, IL-1, and TNF-α. Furthermore, 16S rRNA analysis showed that the Shannon and Simpson indices decreased at 48 h after Pb exposure, and the abundance of pathogenic bacteria (Erysipelotrichaceae, Weeksellaceae, and Vibrionaceae) increased after Pb exposure. In addition, the correlation network analysis found that Proteobacteria were negatively correlated with Firmicutes and positively correlated with Bacteroidetes. Functional prediction analysis of bacteria speculated that the change in intestinal microbiota led to the PPAR signaling pathway and peroxisome function of the intestine of crucian carp was increased, while the immune system and membrane transport function were decreased. Finally, canonical correlation analysis (CCA) found that there were correlations between the intestinal microbiota, morphology, antioxidant factors, and immune factors of crucian carp after Pb exposure. Taken together, our results demonstrated that intestinal flora dysbiosis, morphological disruption, oxidative stress, and immune injury are involved in the toxic damage of Pb exposure to the intestinal structure and function of crucian carp. Meanwhile, Pb exposure rapidly increased the abundance of pathogenic bacteria, leading to intestinal disorders, further aggravating the damage of Pb to intestinal structure and function. These findings provide us a basis for the link between gut microbiome changes and heavy metal toxicity, and gut microbiota can be used as biomarkers for the evaluation of heavy metal pollution in future.
RESUMO
Lead (Pb) is one of the most common trace metals in water, and its high concentration in the environment can cause harm to aquatic animals and humans. In the present study, the effects of Pb exposure (3.84 mg/kg) on the morphology, digestive enzyme activity, immune function and microbiota structure of silver carp (Hypophthalmichthys molitrix) intestines within 96 h were detected. Moreover, the correlation between them was analyzed. The results showed that Pb exposure on the one hand severely impaired the intestinal morphology, including significantly shortening the intestinal villi's length, increasing the goblet cells' number, causing the intestinal leukocyte infiltration, and thickening the intestinal wall abnormally, on the other hand, increasing the activity of intestinal digestive enzyme (trypsin and lipase). In addition, the mRNA expressions of structure-related genes (Claudin-7 and villin-1) were down-regulated, and the immune factors genes (IL-8, IL-10 and TNF-α) were up-regulated after Pb exposure. Furthermore, data of the MiSeq sequencing showed that the abundance of membrane transport, immune system function and digestive system of silver carp intestinal microbiota all decreased, while cellular antigens increased. Finally, the canonical correlation analysis (CCA) showed that there were correlations between silver carp's intestinal microbiota and intestinal morphology and immune factors. In conclusion, it is speculated that the entry of Pb into the intestine leads the microbiota dysbiosis, affects the intestinal immunity and digestive function, and further damages the intestinal barrier of silver carp.