Copper affect the multixenobiotic resistance mechanism (MXR) in embryo and larvae of Rhamdia quelen

P-glycoproteins (P-gp) and Multidrug resistance protein (MRP) represent a family of ABC (ATP-binding cassette) transporters responsible for multixenobiotic resistance mechanism (MXR) in aquatic organisms. In the current study the modulation of P-gp and MRP proteins was evaluated in embryo and larvae of Rhamdia quelen fish species exposed to copper. Adult females were exposed by gavage during 60 days to copper (5 mg Cu kg-1) and eggs, embryos, and larvae from exposed and unexposed females were exposed to 30 mg Cu L-1. The activity of ABC transporters was accessed via calcein accumulation assay using the specific inhibitors: Verapamil (P-gp) and MK571 (MRP). P-gp activity was detected in all analyzed stages whereas MRP activity was observed after 36 and 96 hpf. Oocytes from females previously exposed and larvae stages (36 and 96 hpf) accumulated less calcein than no exposed oocytes, showing higher ABC transporters activity. In individuals exposed to copper, a higher inhibitory effect was observed 1 hpf. The modulation of ABC transporter proteins is time dependent throughout the development, and the initial stages are more sensible to copper. These findings highlight the MXR mechanism as a biomarker of pollutant exposure in early stages of development of R. quelen.

Expression Analyses of Genes Related to Multixenobiotic Resistance in Mytilus galloprovincialis after Exposure to Okadaic Acid-Producing Dinophysis acuminate

The mussel Mytilus galloprovincialis is one of the most important aquaculture species in Europe. Its main production problem is the accumulation of toxins during coastal blooms, which prevents mussel commercialization. P-glycoprotein (ABCB1/MDR1/P-gp) is part of the multixenobiotic resistance system in aquatic organisms, and okadaic acid, the main DSP toxin, is probably a substrate of the P-gp-mediated efflux. In this study, the presence and possible role of P-gp in the okadaic acid detoxification process was studied in M. galloprovincialis. We identified, cloned, and characterized two complete cDNAs of mdr1 and mdr2 genes. MgMDR1 and MgMDR2 predicted proteins had the structure organization of ABCB full transporters, and were identified as P-gp/MDR/ABCB proteins. Furthermore, the expression of mdr genes was monitored in gills, digestive gland, and mantle during a cycle of accumulation-elimination of okadaic acid. Mdr1 significantly increased its expression in the digestive gland and gills, supporting the idea of an important role of the MDR1 protein in okadaic acid efflux out of cells in these tissues. The expression of M. galloprovincialismrp2, a multidrug associated protein (MRP/ABCC), was also monitored. As in the case of mdr1, there was a significant induction in the expression of mrp2 in the digestive gland, as the content of okadaic acid increased. Thus, P-gp and MRP might constitute a functional defense network against xenobiotics, and might be involved in the resistance mechanisms to DSP toxins.

Commercial preparations of pesticides exert higher toxicity and cause changes at subcellular level in earthworm Eisenia andrei

Background The environmental impact of pesticides has been an increasingly discussed issue over the last decades. Constant usage of pesticides presents a burden for soil and causes a decrease in its health, including the negative effects on earthworms which are indicators for soil quality. The objective of this research was the assessment of the effects of two insecticides and two herbicides on the earthworm Eisenia andrei. Namely, the following active ingredients and respective commercial preparations were investigated: esfenvalerate (Sumialfa), thiacloprid (Calypso), dimethenamid-p (Frontier) and prosulfocarb (Filon). Lethal concentrations (48 h) of both active ingredient and commercial preparations were determined using the filter paper contact test. Results The results showed that Calypso and Frontier were significantly more toxic than the active ingredient. Therefore, all further measurements were performed after exposure of earthworms to the commercial preparations of the pesticides. Specifically, several enzymatic biomarkers and multixenobiotic resistance activity were assessed. Additionally, a fluorescence-based assay for the determination of oxidative stress was established. Significant changes were detected for catalase, carboxylesterase and multixenobiotic activities after 48-h exposures. Also, a significant change in oxidative stress parameters could be observed for both Calypso and Frontier. Conclusions The obtained results show that commercial preparations can be more toxic than the active ingredients, and the formulations being distributed in the environment can affect earthworms on a molecular level already after short exposures. This emphasizes the importance of a more integrated eco-toxicological assessment of commercial pesticide preparations not to underestimate their effects on the environment.