{"refrec":{"BRefID":73188,"RR":"<b>Sánchez-Fortún, S.; Llorente, M.T.; Castaño, A.</b> (2005). Genotoxic effects of selected biocides on RTG-2 fish cells by means of a modified Fast Micromethod Assay. <i>Aquat. Toxicol. 73(1)</i>: 55-64. <a href=\"https://dx.doi.org/10.1016/j.aquatox.2005.03.002\" target=\"_blank\">https://dx.doi.org/10.1016/j.aquatox.2005.03.002</a>","BEntID":69097,"PublicFlag":1,"CheckedFlag":1,"wosflag":1,"vabbflag":0,"RefStringPartII":". <i>Aquat. Toxicol. 73(1)</i>: 55-64. <a href=\"https://dx.doi.org/10.1016/j.aquatox.2005.03.002\" target=\"_blank\">https://dx.doi.org/10.1016/j.aquatox.2005.03.002</a>","DocTypID":8,"DocType":"Journal article","MarineFlag":0,"FreshFlag":0,"BrackishFlag":0,"TerrestrialFlag":0,"Authorstring":"Sánchez-Fortún, S.; Llorente, M.T.; Castaño, A.","OrigTitleTranslFlag":0,"Authorstringtrunc":"Sánchez-Fortún, S.; Llorente, M.T.; Castaño, A.","Englishabstract":"A sensitive in vitro assay for detecting DNA damage in RTG-2 cells culture is described. This assay employs a dye, PicoGreen double stranded DNA (dsDNA) quantitation reagent, which becomes intensely fluorescent upon binding nucleic acids. The assay includes a simple and rapid 50-min sample lysis in the presence of EDTA, SDS, and high urea concentration at pH 10, followed by time-dependent DNA denaturation at pH 11.6 after NaOH addition. The time course and the extent of DNA denaturation are followed in a microplate fluoresecence reader at room temperature for less than 1 h. Comparative studies between suspension and fixed RTG-2 cells indicated that it is possible to apply this methodology in both cases with good results. Neutral red assay was used for to determine the cellular viability when RTG-2 cultures were exposed to tetrakis(hydroxymethyl) phosphonium chloride (THPC) and benzalkonium chloride (BC), as biocides used in the disinfection of cooling towers. The results obtained by neutral red assay indicate IC<sub>50(48)</sub> values of 0.017 (0.011-0.028) and 2.71 (1.91-3.86) mg/L for tetrakis(hydroxymethyl) phosphonium chloride and benzalkonium chloride, respectively. DNA damage has been evaluated for both disinfectants in RTG-2 culture, by exposure to 1/10-, 1/25-, 1/50-, and 1/100-IC<sub>50(48)</sub> value, and the results obtained indicate a strain scission factor (SSF) of 0.126 ± 0.014, 0.181 ± 0.014, 0.217 ± 0.013, and 0.245 ± 0.013 in cell suspensions, and 0.077 ± 0.019, 0.107 ± 0.014, 0.151 ± 0.014, and 0.202 ± 0.015 in attached cells for tetrakis(hydroxymethyl) phosphonium chloride; while the SSF values for benzalkonium chloride are 0.023 ± 0.009, 0.033 ± 0.017, 0.068 ± 0.012, and 0.088 ± 0.015 in cell suspensions, and 0.033 ± 0.010, 0.044 ± 0.011, 0.080 ± 0.009, and 0.093 ± 0.010 in attached cells. Thus, the assay proposed in this study has made it possible to show DNA damage in RTG-2 cells when exposed to 0.2 (1/100 IC<sub>50(48)</sub>) and 300 (1/10 IC<sub>50(48)</sub>) Hg/L of tetrakis(hydroxymethyl) phosphonium chloride and benzalkonium chloride, respectively. The results obtained indicate that the Fast Micromethod Assay, applied on RTG-2 cell line cultures, is a fast and sensitive method for the early DNA damage detection in the aquatic environment.","AbstractOtherLang":null,"BibLvlCode":"AS","StandardTitle":"Genotoxic effects of selected biocides on RTG-2 fish cells by means of a modified Fast Micromethod Assay","OrigTitleLangCode":"en","OrigTitleLangCodeExtended":"eng","OrigTitleLangID":15,"DateLastModified":{"date":"2026-06-10 01:31:28.629285","timezone_type":1,"timezone":"+02:00"},"UserAccessRight":null,"UserAccID":null,"AuthorKeywords":"DNA damage; RTG-2 cell line; PicoGreen((R)); tetrakis(hydroxymethyl)phosphonium chloride; benzalkonium chloride","OtherDescriptors":null,"Notes":null,"AnaPub":2005,"MonPub":null,"DateUpdate":"2020-10-06","DateCreate":"2005-06-10","SecASFANote":null,"ConfID":null,"PeerRev":1,"VlizCoreFlag":1,"WoScode":"WOS:000229684000005","VABBcode":null,"OpenAcc":0,"DOI":"10.1016/j.aquatox.2005.03.002"},"refs":null,"anarec":{"AnaID":73188,"PubliDate":2005,"Pagination":"55-64","XtraPublOfAnaID":null,"ISBN":null,"Volume":"73","Issue":"1","BRefMon":null,"BRefMonRR":null,"BRefXtra":null,"BRefXtraRR":null,"SerBRefID":42203,"SerRR":"Aquatic Toxicology. 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