Endosulfan-Induced Biomarkers in Japanese Rice Fish (Oryzias latipes) Analyzed by SELDI-TOF-MS

Abstract

The objective of this study was to find and validate estrogen-related biomarkers from plasma proteins in Oryzias latipes after exposure to an estrogen disrupting compound, α-endosulfan. The acute toxicity of α-endosulfan on O. latipes after 96 h of exposure was 13.72, 16.18, and 22.18 μg L-1 for the LC10, LC20, and LC50 values, respectively. To confirm estrogenic disturbance by α-endosulfan, the expression level of vitellogenin in the liver of male fishes was measured at the LC10 value, and it was found to be significantly different from the reference group, confirming the estrogenic effect of endosulfan in this concentration range. Proteinchip® array techniques using a weak cation exchange (CM10) and a strong anion exchange proteinchip (Q10) in conjunction with surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) were used to determine plasma proteins of O. latipes differently expressed in response to endosulfan exposure at LC10 and LC20 concentrations. Analysis of protein profiling of the male fish exposed to α-endosulfan detected 48 significantly different protein peaks and the proteins at m/z 2819, 8462, 8860, and 9462 were significantly different (p<0.05). The protein peaks at m/z 2819, 8860, and 9462 were up-regulated and the peak at m/z 8462 was down-regulated. Therefore, these four differentially expressed proteins could be used as biomarkers to rapidly determine a possible risk of endosulfan on aquatic ecosystems, although these are not necessarily produced as a result of endocrine disruption.

Highly selective biomarkers for pesticides developed in Eisenia fetida using SELDI-TOF MS

“…Thus, it is worthwhile in applying SELDI-TOF MS to aid in the discovery of novel, specific biomarkers for use in environmental situations. SELDI-TOF MS has been used to identify biomarkers that are specific for endosulfan exposure in Japanese rice fish (Oryzias latipes) (Lee et al, 2013).…”

Acute toxicity and gene responses induced by endosulfan in zebrafish (Danio rerio) embryos

“…[19,20] We recently analyzed lipids and small proteins using MALDI-TOF MS/MS and SELDI-TOF MS in chlorpyrifos-and endosulfan-exposed fish samples. [14,15,21] In this study, MALDI-TOF MS/MS identified phospholipids that had been differentially biosynthesized after exposure to chlorpyrifos in Japanese rice fish. [21] In addition, a phospholipid with a mass ion of 556.32 significantly decreased in concentration after exposure to chlorpyrifos in Japanese medaka.…”

“…[41] Endosulfan also induced increased vitellogenin levels in the livers of male Japanese medaka (O. latipes), confirming estrogenic disturbance. [14] to endosulfan in zebrafish embryos (Figure 3). The acacb gene expresses acetyl-CoA carboxylase beta protein in cells, and this protein plays an important role in the carboxylation of acetyl-CoA to form malonyl-CoA.…”

“…[5] Its toxicity to fish has been reported at LC 50 values lower than those of terrestrial animals. [14,15] In tiger earthworms, Eisenia fetida, endosulfan showed acute toxicity with an LC 50 value of 7.7 g L −1 following direct contact. [15] In Japanese medaka (Oryzias latipes), endosulfan exhibited toxicity with an LC 50 value of 22.18 μg L −1…”

“…[14] One of its metabolites, endosulfan sulfate, is more toxic than the parent compound, with LC 50 values ranging from 2.1 to 3.5 μg L −1 for mosquitofish (Gambusia affinis), least killifish (Heterandria formosa), sailfin mollies (Poecilia latipinna), and fathead minnows (Pimephales promelas).…”