One of the main contributors to pharmaceutical pollution of surface waters are non-steroidal anti-inflammatory drugs (NSAIDs) that contaminate the food chain and affect non-target water species. As there are not many studies focusing on toxic effects of NSAIDs on freshwater fish species and specially effects after dietary exposure, we selected rainbow trout (Oncorhynchus mykiss) as the ideal model to examine the impact of two NSAIDs - diclofenac (DCF) and ibuprofen (IBP). The aim of our study was to test toxicity of environmentally relevant concentrations of these drugs together with exposure doses of 100× higher, including their mixture; and to deepen knowledge about the mechanism of toxicity of these drugs. This study revealed kidneys as the most affected organ with hyalinosis, an increase in oxidative stress markers, and changes in gene expression of heat shock protein 70 to be signs of renal toxicity. Furthermore, hepatotoxicity was confirmed by histopathological analysis (i.e. dystrophy, congestion, and inflammatory cell increase), change in biochemical markers, increase in heat shock protein 70 mRNA, and by oxidative stress analysis. The gills were locally deformed and showed signs of inflammatory processes and necrotic areas. Given the increase in oxidative stress markers and heat shock protein 70 mRNA, severe impairment of oxygen transport may be one of the toxic pathways of NSAIDs. Regarding the microbiota, an overgrowth of Gram-positive species was detected; in particular, significant dysbiosis in the Fusobacteria/Firmicutes ratio was observed. In conclusion, the changes observed after dietary exposure to NSAIDs can influence the organism homeostasis, induce ROS production, potentiate inflammations, and cause gut dysbiosis. Even the environmentally relevant concentration of NSAIDs pose a risk to the aquatic ecosystem as it changed O. mykiss health parameters and we assume that the toxicity of NSAIDs manifests itself at the level of mitochondria and proteins.
- MeSH
- antiflogistika nesteroidní metabolismus MeSH
- biologické markery metabolismus MeSH
- chemické látky znečišťující vodu * metabolismus MeSH
- diklofenak metabolismus MeSH
- dysbióza MeSH
- ekosystém MeSH
- epidemický výskyt choroby MeSH
- ibuprofen metabolismus toxicita MeSH
- kyslík metabolismus MeSH
- léčivé přípravky metabolismus MeSH
- messenger RNA metabolismus MeSH
- Oncorhynchus mykiss * metabolismus MeSH
- oxidační stres MeSH
- proteiny tepelného šoku HSP70 metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- střevní mikroflóra * MeSH
- voda metabolismus MeSH
- zánět chemicky indukované MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
In this study we have compared protein secretion in the wild type of S. Typhimurium and the rfaC mutant. We found out that the rfaC mutant was defective in protein secretion. In addition, the rfaC mutant was defective in its invasion into an IPEC-J2 porcine epithelial cell line and also in motility in semisolid agar. Consistent with this, reduced flagella numbers were observed in the rfaC mutant. In the rfaC mutant, there were no defects in flagellin expression as detected by western blot and immune electron microscopy which demonstrated equal amounts of flagellin in the cytoplasm of both the rfaC mutant and the wild-type S. Typhimurium. However, in the wild-type strain only, the flagellin was assembled to spatially restricted areas on the inner side of cytoplasmic membrane. The oligosaccharide core of LPS is therefore required for the assembly of flagella and T3SS secretion machinery followed by protein secretion.
- MeSH
- bakteriální sekreční systémy MeSH
- buněčné linie MeSH
- cytoplazma chemie MeSH
- epitelové buňky mikrobiologie MeSH
- flagelin biosyntéza sekrece MeSH
- flagella metabolismus MeSH
- imunoelektronová mikroskopie MeSH
- lipopolysacharidy chemie MeSH
- mutace MeSH
- prasata MeSH
- Salmonella enterica genetika metabolismus ultrastruktura MeSH
- sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
