Journal of Environmental Biology

In vivo and in vitro toxicities of diethyl phthalate to flounder fish Paralichthys olivaceus and its gill cell line (FG cells)

Qin Xiao¹*, Daizong Li¹, Ran Guo¹, Lei Zheng², Xinlong An¹ and Zhaochun Zeng¹*

¹College of Ocean, Hebei Agricultural University, Qinhuangdao, 066 003, China

²College of Water Sciences, Beijing Normal University, Beijing, 100875, China

*Corresponding Author E-mail: qinxiao669@163.com

Key words

Diethyl phthalate

FG cell line

Paralichthys olivaceus

Toxicity 

Publication Data

Paper received : 22.09.2016?????????????????? Revised received : 19.04.2017????????????????????? Re-revised received : 10.07.2017 Accepted : 21.08.2017???????? ??

Abstract

Aim: Diethyl phthalate (DEP) as a plasticizer and softener of daily supplies, is immensely released in the aquatic environment. The study was undertaken in order to evaluate the toxicity of DEP using the marine flounder (Paralichthys olivaceus) and its gill cell line (FG cells) in vivo and in vitro.

Methodology: In vitro toxicity of DEP to FG cells was determined by neutral red uptake assay and methyl thiazolyl tetrazolium assay, along with observing the change of cellular morphology by a phase contrast inverted microscope. In vivo toxicity of DEP to flounder fish was evaluated by calculating the median lethal concentration (LC₅₀) in semi-static condition, observing the histopathological change in liver and kidney of flounders by staining with hematoxylin and eosin, and determining the biochemical parameters using an AEROSET automatic biochemical analyzer. ????

Results: DEP was acutely toxic to FG cells for 24hr in the range of 200 μg ml^-1-800μg ml^-1. 50% inhibition of cell proliferation (IC₅₀) of DEP for 24 hr were 536.76 μg ml-1 and 573.84 μg ml-1 for NR uptake and MTT assay, respectively. Meanwhile, the FG cellular morphology was also negatively altered with EDP concentration increasing. In semi-static condition, the median lethal concentration (LC₅₀) of DEP for 96 hr was 31.92 μg

ml^-1 for Paralichthys olivaceus. DEP also caused various extents of histological structures damages in livers and kidneys when fish were exposed to 5, 10 and 15 μg ml^-1 DEP, as evidenced by the necrosis of hepatocytes, the disorder of hepatic sinusoids, the degradation of tubular epithelial cells and multiple hemorrhages in kidneys. Meanwhile, the levels of alanine aminotransferase and lactate dehydrogenase were enhanced at the same concentration DEP treatments. ????

Interpretation: The study suggests that FG cell line can be a short term bioassay model for screening the potential toxicity of the pollutant, while living fish are a necessary model for studying DEP toxicity. This research will provide significant references for monitoring DEP pollution.

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