Processes involved in biochemical response to pesticides by lizard Podarcis siculus (Rafinesque-Schmaltz, 1810) - A field study.

Although reptiles are non-target organisms of pesticide applications, their ecological niche and trophic role suggest that the use of these compounds in agriculture can have toxicological effects on them. Our recent field study on Italian wall lizard Podarcis siculus in hazelnut orchards evidenced that the use of pesticides-mixtures, consisting of thiophanate-methyl (TM), tebuconazole (TEB), deltamethrin (DM), lambda-cyhalothrin (LCT), besides copper sulphate, induced an increase of the total antioxidant capacity toward hydroxyl radicals and caused DNA damage; however, it did not cause neurotoxicity, and did not induce the glutathione-S-transferases' activities. These results raised some questions which were answered in this study by carrying out analyses on 4 biomarkers and 5 chemical substances in the tissues of non-target organisms coming from treated fields: cytochrome P450, catalase, total glutathione, and malondialdehyde, TM, TEB, DM, LCT and Cu. Our results highlighted a partial accumulation of different chemicals, the involvement of two important mechanisms of defence, and some cellular damages after exposure to the considered pesticides. In details, 1) LCT and DM were not accumulated in lizard muscle, copper remained at basal levels, whereas TM and TEB were uptaken with a partial metabolization of TM; 2) the cytochrome P450 and the catalase were involved in lizard biochemical responses to pesticides-mixtures used for "conventional" farming treatment; 3) "conventional" treatment with pesticides caused damage to lipids, besides DNA, probably related to the excess of hydroxyl radicals.

Biological responses in pesticide exposed lizards (Podarcis siculus).

The release of contaminants as herbicides, fungicides and insecticides into the environment has been listed as one of the six major contributors to the global decline of reptiles. Although reptiles may face severe risk from contaminants due to their ecology and physiology, they are currently less studied than other vertebrate groups. In the present work, we investigated if and how different types of field treatment (conventional and organic) affected the health status of Italian wall lizard (Podarcis siculus) individuals in central Italy. We chose a multi-biomarker approach that evaluated the biological responses of lizards to the treatment by means of AChE activity in the nervous system, biotransformation enzymes activities and oxidative stress in the liver, micronuclei frequency measured in the erythrocytes, and rate of intestinal parasitic infection. Our findings showed evidence of effects of treatment in conventional areas and between sexes with significant oxidative stress due to hydroxyl radicals, that caused DNA damage. No difference of intestinal parasite infections was found among treatments. Podarcis siculus seems to be a good bioindicator in ecotoxicological studies and potentially in risk assessment of pesticides, although further analyses in laboratory and in the field are needed to achieve more accurate quantification of specific pesticide effects in relation to known exposure history and to understand if other mechanisms were involved in the toxicity and detoxification process of pesticides for this species.

Biological responses of the polychaete Hediste diversicolor (O.F.Müller, 1776) to inorganic mercury exposure: A multimarker approach.

Mercury (Hg) is a global priority pollutant given its relevance in terms of environmental damage and threat to human health. Its ecotoxicity was tested using the benthic keystone species Hediste diversicolor as target species. After 10 days of exposure to different levels of inorganic Hg (10 and 50 µg L-1), bioaccumulation and a wide range of biological responses were evaluated at different biological levels, including biomarkers of exposure, neurotoxicity, oxidative stress, genotoxicity and cytochemistry. In controlled laboratory conditions, Hg was taken up by H. diversicolor in a dose-response manner and caused a range of biological responses, including oxidative stress (GSTs, GPx, GSH-2GSSG, and TOSCA), neurotoxicity (AChE), and cellular damages at the membrane level (LFs, NLs, Ca2+-ATPase); however, it did not cause significant DNA damage or mortality. This study confirms the capability of H. diversicolor to tolerate high levels of metals and clarifies the mechanisms underlying the damage caused by waterborne Hg and the defense mechanisms, activated in this species. In particular, detoxification of the inorganic form of Hg in this species was found to be strongly related to glutathione expression and several antioxidant enzymes of the antioxidant system. This process also efficiently minimized negative effects on DNA and prevented death, but was not sufficient to avoid neurotoxicity and some cellular damages, mainly at the intestinal level.

An ecotoxicological protocol with caged mussels, Mytilus galloprovincialis, for monitoring the impact of an offshore platform in the Adriatic Sea.

An ecotoxicological protocol with caged mussels, Mytilus galloprovincialis, was developed to evaluate the potential impact of an offshore gas platform in the central Adriatic Sea. Reference organisms were collected on a seasonal basis from an unpolluted site and transplanted for four weeks in both the sampling area and to the investigated platform. Chemical analyses of trace metals in mussel tissues were integrated with a multi-biomarker approach for the early detection of biological responses at several cellular targets. Induction of metallothioneins, peroxisomal proliferation and activity of acetylcholinesterase were measured as markers for specific classes of chemicals. Special attention was given to oxyradical metabolism and appearance of oxidative-mediated toxicity to reveal a more general onset of cellular disturbance. In addition to individual antioxidants (superoxide dismutase, catalase, glutathione S-transferases, glutathione reductase, Se-dependent and Se-independent glutathione peroxidases, and levels of total glutathione), the total oxyradical scavenging capacity (TOSC) allowed a quantification of the overall capability to neutralize specific forms of intracellular reactive oxygen species (ROS; i.e. peroxyl and hydroxyl radicals). Cellular damages were evaluated as lysosomal destabilization (membrane stability, accumulation of lipofuscin and neutral lipids), lipid peroxidation products (malondialdehyde) and DNA integrity (strand breaks and micronuclei); the air survival test was finally applied to evaluate the overall physiological condition of mussels. Concentration of trace metals (As, Ba, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Zn) revealed only limited variations in transplanted mussels during various experimental periods and such changes appeared partly related to natural fluctuations. Among biological responses, variations of antioxidants and lysosomal stability were confirmed as sensitive early warning signals for biological disturbance of both natural and anthropogenic origin. The presented protocol with caged mussels allowed marked biological effects caused by the investigated platform to be excluded, and represented a useful approach that is easy to extend for monitoring the impact of offshore activities in the Adriatic sea.