Predicting fish acute toxicity of chemicals in vitro is an attractive alternative method to the conventional approach using juvenile and adult fish. The rainbow trout (Oncorhynchus mykiss) cell line assay with RTgill-W1 cells has been designed for this purpose. It quantifies cell viability using fluorescent measurements for metabolic activity, cell- and lysosomal-membrane integrity on the same set of cells. Results from over 70 organic chemicals attest to the high predictive capacity of this test. We here report on the repeatability (intralaboratory variability) and reproducibility (interlaboratory variability) of the RTgill-W1 cell line assay in a round-robin study focusing on 6 test chemicals involving 6 laboratories from the industrial and academic sector. All participating laboratories were able to establish the assay according to preset quality criteria even though, apart from the lead laboratory, none had previously worked with the RTgill-W1 cell line. Concentration-response modeling, based on either nominal or geometric mean-derived measured concentrations, yielded effect concentrations (EC50) that spanned approximately 4 orders of magnitude over the chemical range, covering all fish acute toxicity categories. Coefficients of variation for intralaboratory and interlaboratory variability for the average of the 3 fluorescent cell viability measurements were 15.5% and 30.8%, respectively, which is comparable to other fish-derived, small-scale bioassays. This study therefore underlines the robustness of the RTgill-W1 cell line assay and its accurate performance when carried out by operators in different laboratory settings.
- MeSH
- aniliny toxicita MeSH
- buněčné linie MeSH
- laboratoře MeSH
- Oncorhynchus mykiss MeSH
- reprodukovatelnost výsledků MeSH
- testy akutní toxicity metody MeSH
- viabilita buněk účinky léků MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Selection of appropriate test species is a critical issue when assessing effects of environmental contamination on fish because the ecological relevance of commonly used test species might be restricted due to their exotic origin. In the present study, a European freshwater fish with frequent occurrence in agricultural areas is suggested as a potential alternative: the European weatherfish (Misgurnus fossilis). Its suitability for acute embryo toxicity tests (FET) was investigated with regard to practical implementation, sensitivity to contaminants and tolerance against environmental conditions of concern. For this purpose, weatherfish embryos were exposed (72h) to the reference substance 3,4-dichloroaniline (DCA) in three independent tests. Furthermore, the effects of dissolved oxygen (DO) deficiency on weatherfish embryos were studied to evaluate their suitability e.g. for sediment bioassays. Obtained results revealed that the sensitivity of weatherfish embryos towards DCA (72 h-EC50=0.52mg/l; 72 h-LC50=0.71mg/l) was highest compared to other species and three times higher than that reported for the commonly used zebrafish (Danio rerio). Even though knowledge of DO requirements during the embryonic period of European fish species is scarce, weatherfish can be stated as one of the most tolerant native species (LC90 for DO=0.53mg/l after 48h exposure plus 72h post-exposure). Its high ecological relevance for Europe, the particular sensitivity towards DCA and high tolerance against DO depletion highlight the potential of weatherfish as additional species for toxicity testing.
2-Methoxyaniline (o-anisidine) is an industrial and environmental pollutant causing tumors of urinary bladder in rodents. Here, we investigated the formation and persistence of DNA adducts in the Wistar rat. Using the (32)P-postlabeling method, three o-anisidine-derived DNA adducts were found in several organs of rats treated with a total dose of 0.53 mg o-anisidine/kg body wt (0.15, 0.18, and 0.2 mg/kg body wt ip in the first, second, and third day, respectively), of which the urinary bladder had the highest levels. At four posttreatment times (1 day, 13 days, 10 weeks, and 36 weeks), DNA adducts in bladder, liver, kidney, and spleen of rats were analyzed to study their persistence. In all time points, the highest total adduct levels were found in urinary bladder (39 adducts per 10(7) nucleotides after 1 day and 15 adducts per 10(7) nucleotides after 36 weeks) where 39% adducts remained. In contrast to the urinary bladder, no persistence was detected in other organs. All three DNA adducts were identified as deoxyguanosine adducts. When deoxyguanosine was reacted with the oxidative metabolite of o-anisidine, N-(2-methoxyphenyl)hydroxylamine, three adducts could be separated by high-performance liquid chromatography (HPLC) and were identified by mass spectroscopy and/or nuclear magnetic resonance spectrometry. All adducts are products of the nitrenium/carbenium ions, the reactive species generated from N-(2-methoxyphenyl)hydroxylamine. The major adduct was identified to be N-(deoxyguanosin-8-yl)-2-methoxyaniline. Using cochromatography on HPLC, this adduct was found to be identical to the major adduct generated by activation of o-anisidine in vitro and in vivo.
- MeSH
- adukty DNA metabolismus MeSH
- aniliny metabolismus toxicita MeSH
- časové faktory MeSH
- deoxyguanosin metabolismus MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- hydroxylaminy metabolismus MeSH
- játra účinky léků metabolismus MeSH
- krysa rodu rattus MeSH
- látky znečišťující životní prostředí metabolismus toxicita MeSH
- ledviny účinky léků metabolismus MeSH
- magnetická rezonanční spektroskopie MeSH
- močový měchýř účinky léků metabolismus MeSH
- potkani Wistar MeSH
- slezina účinky léků metabolismus MeSH
- tkáňová distribuce MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cieľom článku je podať informáciu o možnom riziku vzniku para-chlóranilínu (PCA), ktorý je rozkladovým produktom chlórhexidínu (CHX) a zlúčeninou s toxickým a karcinogénnym účinkom. Vznik PCA je možným následkom používania chlórhexidínu, ktorý je v bežnej praxi široko používaný ako dezinfekčný prostriedok pri odstraňovaní zubného povlaku a v endodontickej terapii devitalizovaných - pulpy zbavených zubov, na redukciu mikrobiálnej flóry.
The purpose of this article is to report on potential risk of formation of para-chloroaniline (PCA), a breakdown product of chlorhexidine (CHX) a toxic and carcinogenic compound. The generation of PCA is a potential consequence of the use of chlorhexidine which has been widely used as an antiseptic agent for routine dental plaque control and irrigation of root canals in endodontic treatment of devitalised, pulpless teeth, for reduction of microbiota.
- Klíčová slova
- para-chloranilin, rozkladový produkt chlorhexidinu, PCA, 4-chloranilin,
- MeSH
- aniliny farmakokinetika terapeutické užití toxicita MeSH
- chlorhexidin škodlivé účinky MeSH
- karcinogeny farmakologie metabolismus MeSH
- lékové interakce MeSH
- lidé MeSH
- methemoglobinemie chemicky indukované MeSH
- zubní dezinficiencia chemie škodlivé účinky MeSH
- zuby neživé farmakoterapie krev ošetřování MeSH
- Check Tag
- lidé MeSH
o-Aminophenol and N-(2-methoxyphenyl)hydroxylamine are human metabolites of the industrial and environmental pollutant and bladder carcinogen 2-methoxyaniline (o-anisidine). The latter one is also a human metabolite of another pollutant and bladder carcinogen, 2-methoxynitrobenzene (o-nitroanisole). Here, we investigated the ability of rat hepatic micro- somes to metabolize these metabolites. N-(2-methoxyphenyl)hydroxylamine is metabolized by rat hepatic microsomes to o-aminophenol and predominantly o-anisidine, the parent carcinogen from which N-(2-methoxyphenyl)hydroxylamine is formed. In addition, two N-(2-methoxyphenyl)hydroxylamine metabolites, whose exact structures have not been identified as yet, were generated. On the contrary, no metabolites were found to be formed from o-aminophenol by rat hepatic microsomes. Whereas N-(2-methoxyphenyl)hydroxylamine is responsible for formation of three deoxyguanosine adducts in DNA, o-aminophenol seems to be a detoxication metabolite of N-(2-methoxyphenyl)hydroxylamine and/or a parental carcinogen, o-anisidine; no o-aminophenol-derived DNA adducts were found after its reaction with microsomal cytochromes P450 and peroxidases.
- MeSH
- aminofenoly metabolismus toxicita MeSH
- aniliny metabolismus toxicita MeSH
- biotransformace MeSH
- hydroxylaminy metabolismus toxicita MeSH
- játra metabolismus MeSH
- karcinogeny životního prostředí metabolismus toxicita MeSH
- krysa rodu rattus MeSH
- metabolická clearance MeSH
- metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- MeSH
- adukty DNA analýza MeSH
- aniliny toxicita MeSH
- finanční podpora výzkumu jako téma MeSH
- jaterní mikrozomy MeSH
- karcinogeny životního prostředí toxicita MeSH
- krysa rodu rattus MeSH
- modely u zvířat MeSH
- systém (enzymů) cytochromů P-450 metabolismus MeSH
- testy karcinogenity MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- MeSH
- aniliny chemie toxicita MeSH
- antiinfekční látky MeSH
- Bacteria účinky léků MeSH
- chinazoliny chemie toxicita MeSH
- Publikační typ
- srovnávací studie MeSH
