Among all classes of nanomaterials, silver nanoparticles (AgNPs) have potentially an important ecotoxicological impact, especially in freshwater environments. Fish are particularly susceptible to the toxic effects of silver ions and, with knowledge gaps regarding the contribution of dissolution and unique particle effects to AgNP toxicity, they represent a group of vulnerable organisms. Using cell lines (RTL-W1, RTH-149, RTG-2) and primary hepatocytes of rainbow trout (Oncorhynchus mykiss) as in vitro test systems, we assessed the cytotoxicity of the representative AgNP, NM-300K, and AgNO3 as an Ag+ ion source. Lack of AgNP interference with the cytotoxicity assays (AlamarBlue, CFDA-AM, NRU assay) and their simultaneous application point to the compatibility and usefulness of such a battery of assays. The RTH-149 and RTL-W1 liver cell lines exhibited similar sensitivity as primary hepatocytes towards AgNP toxicity. Leibovitz’s L-15 culture medium composition (high amino acid content) had an important influence on the behaviour and toxicity of AgNPs towards the RTL-W1 cell line. The obtained results demonstrate that, with careful consideration, such an in vitro approach can provide valuable toxicological data to be used in an integrated testing strategy for NM-300K risk assessment.
Normal functioning of the endocrine system is essential for the proper development and reproduction of animals. Substances interfering with its homeostasis are called endocrine disruptors (EDs) and may represent a risk for the health of the organism. One of the mechanisms of endocrine disruption that has attracted great attention in recent years concerns alterations in the normal functioning of the estrogen receptor (ER), but far less attention has been paid to those substances interfering with the thyroid axis, which, in fish, plays several critical roles in a variety of biological functions. In aquaculture, feedstuffs can be a source of hormones or persistent pollutants which act as potential EDs. In this study, the main purpose was to assess the possible estrogenic and thyrogenic activities of 32 commercial fish feeds. For the assessment of estrogenicity, a new estrogen receptor specific reporter gene assay using sea bass ERα (sbER α) was developed and validated. Potential thyroidal disruption was screened with a cell line permanently transfected with luciferase as reporter gene under the control of avian (av) thyroid receptor α (THRα). The results obtained showed that 11 and 18 out of 32 assayed feeds were able to activate the sbERα or the avTHRα1, respectively. The present study is pioneer in demonstrating thyrogenic activity in fish diets commercially available and widely used in aquaculture. Given that maintaining the homeostasis in the endocrine system is critical for the proper development and reproduction of fish, any estrogenic or thyrogenic activity caused by the feedstuffs should be taken into account with regards to its potential impact on farmed fish. Dr. Mosconi has always worked on fish physiology and in the last years has moved to the study of the interaction of pollutants with the reproductive physiology of fish.Maren Ortiz-Zarragoitia Dr. Professor, Zoology and Cell Biology, University of the Basque Country maren.ortiz@ehu.es Dr. Ortiz-Zarragoitia has a wide expertise in the field of ecotoxicology. Since the article we are sending deals with estrogenic and thyrogenic activities of fish feed, Dr Ortiz-Zarragoitia can act as reviewer due to his knowledge about the effect of pollutants on the endocrine system of fish. LAP Hoogenboom Dr. RIKILT Inst Food Safety ron.hoogenboom@wur.nl Dr. Hoogenboom has a broad expertise in the field of residue contaminants in feed and food. He has performed a variety of studies related with the detection of very different contaminants in feed, including estrogenic substances, growth-promoting agents, veterinary drugs and environmental dioxin-like substances.The estrogenicity and thyrogenic activity of fish feed extracts was assessed by means of cell lines containing the estrogen receptor or the thyroid receptor and reporter genes under the control of the corresponding receptor. For the assessment of estrogenic activities a new cell line containing the sea bass estrogen receptor has been developed. Although some estrogenic activity could be detected, strikingly a ve...
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