Induction to oxidative stress by saxitoxin investigated through lipid peroxidation in Neuro 2A cells and Chlamydomonas reinhardtii alga

Saxitoxin (STX) is a cyanotoxin, which can cause neurotoxic effects and induce ecological changes in aquatic environments, a potential risk to public and environmental health. Many studies of cytotoxicity on animal cells and algae have been performed, although few compare the toxic effects between the two models. In this sense, we investigated the oxidative stress induced by STX (0.4-3.0 nM) in two different cellular models: Neuro-2A (N2A) cells and Chlamydomonas reinhardtii alga by quantification of malondialdehyde (MDA) levels as indicative of lipid peroxidation (LPO). Also was evaluated the antioxidant defense of these cells systems after exposure to STX by the addition of antioxidants in N2A cells culture, and by the measure of antioxidants enzymes activity in C. reinhardtii cells. The MDA levels of N2A cells increased from 15% to 113% for 0.4 and 3.0 nM of STX, respectively, as compared to control. Superoxide-dismutase and catalase did not appear to protect the cell from STX effect while, in cells treated with vitamin E, the rates of MDA production decreased significantly, except for higher concentrations of STX. No MDA productions were observed in algal cells however some effects on antioxidant enzymes activity were observed when algae were exposed to 3.0 nM STX. Our results indicate that the concentrations of STX that may induce oxidative stress through LPO are different in animal and phytoplankton communities. A combination of algal and animal bioassays should be conducted for reliable assessment of oxidative stress induced by STX.     

Ability of Poplar (Populus spp.) to Detoxify Chloroacetanilide Herbicides

The ability of poplar trees to resist chemical stress caused by chloroacetanilide herbicides was studied. Detached leaves of Lombardy poplar (Populus nigra L.) were exposed to seven chloroacetanilide herbicides via uptake through the cut petiole. The leaves showed high tolerance against the phytotoxicity of these compounds. Their tolerance was further enhanced by treatment with 2-oxothiazolidine-4-carboxylic acid (OTC, a precursor of the amino acid cysteine). High levels of glutathione (GSH) and GSH S-transferase (GST) activity were detected in poplar leaves. Treatments with chloroacetanilide herbicides left the GSH content in the leaves unchanged but strongly induced the GST activity. In contrast, in OTC-treated leaves increased GSH contents were measured, but GST activities remained unchanged. It therefore appears that a highly active and inducible GSH-conjugation Phase II detoxification system in their leaves may make poplar trees useful for phytoremediation of soils contaminated with chloroacetanilide herbicides. Based on these findings a phytoremediation project using different poplar hybrids at a site heavily polluted with such compounds in Hungary has been launched.     

Effect of municipal waste water effluent upon the expression of Glutathione S-transferase isoenzymes of brine shrimp Artemia

Multiple isoenzymes of the detoxification enzyme family Glutathione S-transferase are expressed in the brine shrimp Artemia. The number of the major ones detected in crude extract by means of chromatofocusing varied between three and four, depending on the age. Two isoenzymes, one alkaline and one neutral (with corresponding isoelectric points of 8.5 and 7.2) appear to be dominant in all three developmental stages studied, (24, 48, and 72 h after hatching). Culturing Artemia for 48 h after hatching, in artificial sea water prepared by municipal wastewater effluent resulted to significant alterations of the isoenzyme profile. In comparison to organisms cultured for the same period of time in artificial sea water prepared by filtered tap water, the expression of the alkaline isoenzyme decreased by 62% while that of the neutral isoenzyme increased by 58%. Furthermore, the enzyme activity of the major isoenzyme of the acidic area increased by more than two folds. It is worth mentioning that although the specific activity of the total enzyme in the whole body homogenate was elevated, no statistically significant alteration of the Km value was observed. These findings suggest that study of the isoenzyme profile of Glutathione S-transferase may offer high sensitivity in detecting environmental pollution and needs to be further investigated.     

Screening for antioxidant and detoxification responses in Perna canaliculus Gmelin exposed to an antifouling bioactive intended for use in aquaculture

Polygodial is a drimane sesquiterpene dialdehyde derived from certain terrestrial plant species that potently inhibits ascidian metamorphosis, and thus has potential for controlling fouling ascidians in bivalve aquaculture. The current study examined the effects of polygodial on a range of biochemical biomarkers of oxidative stress and detoxification effort in the gills of adult Perna canaliculus Gmelin. Despite high statistical power and the success of positive controls, the antioxidant enzymes glutathione reductase (GR), glutathione peroxidase (GPOX), catalase (CAT), and superoxide dismutase (SOD); thiol status, as measured by total glutathione (GSH-t), glutathione disulphide (GSSG), and GSH-t/GSSG ratio; end products of oxidative damage, lipid hydroperoxides (LHPO) and protein carbonyls; and detoxification pathways, represented by GSH-t and glutathione S-transferase (GST), were unaffected in the gills of adult P. canaliculus exposed to polygodial at 0.1 or 1 × the 99% effective dose in fouling ascidians (IC₉₉). Similarly, GR levels, thiol status, and detoxification activities were unaffected in mussels exposed to polygodial at 10 × the IC₉₉, although GPOX, CAT, and SOD activities increased. However, the increases were small relative to positive controls, no corresponding oxidative damage was detected, and this concentration greatly exceeds effective doses required to inhibit fouling ascidians in aquaculture. These findings compliment a previous study that established the insensitivity to polygodial of P. canaliculus growth, condition, and mitochondrial functioning, providing additional support for the suitability of polygodial for use as an antifouling agent in bivalve aquaculture.     

Integrated assessment of biochemical responses in Mediterranean crab (Carcinus maenas) collected from Monastir Bay, Tunisia

The biochemical response of Mediteranean Crab (Carcinus maenas) collected at five stations of Monastir Bay and from Kuriat station as control was studied using a set of complementary biomarkers. The catalase, glutathione S-transferase, lactate dehydrogenase, acetycholinesterase activities; and metallothionein and malonediladehyde levels in gills were evaluated. Results revealed differences among sites in relation to each specific biomarker. Hence, a suite of biomarkers can be used to discriminate sampling sites according to types of pollution, reflecting differing conditions of anthropogenic impact. Based on Integrated Biomarker Response, the highest values and critical biochemical alteration were observed at Khniss and Ksibat in response to urban and industrial discharges and the lowest IBR value was found at reference site. The current study has shown clearly that a biomarker-based index is usefulness tool in the monitoring Tunisian coast using C. maenas as sentinel specie. Further studies in progress to investigate the seasonal variations of IBR levels and its relationship to pollutants concentrations in the sediment, gills and digestive gland of Carcinus maenas from Monastir Bay.     

Bioaccumulation and function analysis of glutathione S-transferase isoforms in Manila clam Ruditapes philippinarum exposed to different kinds of PAHs

This study analyzed the function of different glutathione S-transferase(GST) isoforms and detoxification metabolism responses in Manila clam, Ruditapes philippinarum, exposed to 4 kinds of polycyclic aromatic hydrocarbons(PAHs) single, and their mixtures for 15 days under laboratory conditions. 13 kinds of GSTs in R. philippinarum were classified, and the results of tissue distribution indicated that 12 kinds of GSTs(except GST sigma 3) expressed most in digestive glands. We detected the m RNA e...     

近江牡蛎4种类型GST基因cDNA全序列的克隆与分析

为从分子水平上研究不同类型谷胱甘肽S-转移酶(Glutathione S-transferase,GST)基因在近江牡蛎(Crassostrea ariakensis)等贝类体内代谢去毒过程中的作用,采用RT-PCR及RACE法,分离、克隆得到近江牡蛎肝脏4种类型GST基因cDNA全序列.序列分析结果表明,mu、pi、omega、sigma4种类型GST基因cDNA全长分别为970bp、773bp、999bp、1277bp,分别编码215、207、242、203个氨基酸.构建系统进化树发现,除太平洋牡蛎(Crassostrea gigas)pi型GST外,所有类型GSTs序列都能各自聚集成一簇.     

Effect of Cd on GSH and GSH-related enzymes of Chlamydomonas sp. ICE-L existing in Antarctic ice

Glutathione(GSH) and GSH-related enzymes play a great role in protecting organisms from oxidative damage. The GSH level and GSH-related enzymes activities were investigated as well as the growth yield and malonyldialdehyde(MDA) content in the Antarctic ice microalga Chlamydomonas sp. ICE-L exposure to the different cadmium concentration in this paper. The results showed that the higher concentration Cd inhibited the growth of ICE-L significantly and Cd would induce formation of MDA. At the same time, it is clear that GSH level, glutathione peroxidases(GPx) activity and glutathione S-transferases(GST), activity were higher in ICE-L exposed to Cd than the control. But GR activity dropped notably when ICE-L were cultured in the medium containing Cd. Increase of GSH level, GPx and GST activities acclimate to oxidative stress induced by Cd and protect Antarctic ice microalga Chlamydomonas sp. ICE-L from toxicity caused by Cd exposure. These parameters may be used to assess the biological impact of Cd in the Antarctic pole region environment.