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.

European legislative framework supports a multidisciplinary strategy of environmental monitoring because the environment is a complex system of abiotic and biotic interactions, and it should not be studied and protected by looking at one single aspect. The resulting heterogeneous data request to be carefully processed, and the application of Weight of Evidence (WOE) approaches is, thereby, an integrated validated tool. In this perspective, the present study aims to: (i). apply a specific model (Sediqualsoft) based on the WOE approach for processing multidisciplinary data related to four Lines Of Scientific Evidence (LOEs: chemical analyses, ecotoxicological bioassays, bioaccumulation tests and biomarkers) regarding sediments from an area of the Adriatic Sea; (ii). evaluate the usefulness of this specific integrated approach to estimate the potential environmental hazard due to the presence of gas production platforms respect to the traditional approach of sediment chemical characterization. This latter recognized a more contaminated area within 100 m of the platforms in which the Sediqualsoft model showed the presence of a chemical hazard, ranging from moderate to severe, and identified the contaminants (e.g., some metals, benzo(a)pyrene and acenaphthene) most responsible for it. A significant hazard also appeared in some of the sampled stations by analyzing the LOEs dedicated to the biological responses. The choice of different reference values (regulatory limits, threshold values or concentrations measured in the control area) influenced only the chemical hazard but not the overall integration with other LOEs, showing a moderate hazard for the majority of stations. Here, the concentrations measured in a control area are firstly proposed as possible reference values in Sediqualsoft model applications; this could be of particular relevance when Sediment Quality Guidelines are not available for all the measured substances. Moreover, the limitations of a conventional pass-to-fail approach or worst-case scenario were overcoming interpreting whole chemical and ecotoxicological results. All data analyzed and discussed confirm Sediqualsoft as a suitable tool for processing environmental data, including those first processed here on a monitoring scenario of gas platforms that discharge Produced Water into the sea.

During 2006–2008, a pipeline was buried in Vallona lagoon in the Northern Adriatic Sea (Italy). A Before-During-After environmental monitoring programme was scheduled to monitor possible alterations. Bioaccumulation of metal(loid)s, BTs (butyltins) and HMW-PAHs (High Molecular Weight Polycyclic Aromatic Hydrocarbons), and biological responses (Condition index, air Survival—LT_50, Acetylcholinesterase, Micronuclei—MN, acyl-CoA oxidase, catalase, malondialdehyde—MDA, and the total oxyradical scavenging capacity—TOSCA) were investigated in Manila clams (Ruditapes philippinarum) from November 2005 to June 2015. In opera (IO) results showed higher levels of HMW-PAHs (73 ± 13 ng/g), BTs (90 ± 38 ng Sn/g) and increasing levels of Pb (6.7 ± 0.7 mg/kg) and Zn (73.6 ± 6.08 mg/kg) probably linked to works. Other contaminant alterations, especially metal(loid)s, before (AO) and after (PO) the burial, were attributed to a general condition of the area and mostly unrelated to works. In addition, LT₅₀, MN and TOSCA showed alterations, probably due to hotspots occurring in IO. TOSCA and MDA increases, right after the burial, were considered delayed responses of IO, whilst other biological responses detected later were connected to the general condition of the area. Comparisons between results of Principal Component Analyses (PCAs) highlighted partial overlapping of AO and IO, whilst PO differed only for contaminants. Visual correlations between PCAs highlighted the biomarkers’ latter response.

Conventional methods to analyze biochemical processes related to contaminant toxicity usually require the sacrifice of animals to collect tissues and organs. However, for ethical reasons and especially for endangered species, non- or minimal-invasive methods should be preferred. Among vertebrates, reptiles show a general decline worldwide and therefore the use of non- or minimal-invasive methods to measure some biochemical processes in these animals are encouraged. It is well known that most lizards use a common safety behavior implying the natural loss of tail in the case of predation events. Therefore, if common analyses testing contaminant toxicity could be performed in tail tissue, this method, not implying the sacrifice of the animals, could be considered as a good minimal-invasive method. The aim of this study is to test on wild Italian wall lizard Podarcis siculus the use of tail to detect a large set of biomarkers including oxidative stress (TOSCAROO, TOSCAOH, CAT, tGSH, MDA), biotransformation processes (EROD, GSTs) and neurotoxicity (AChE, BChE). All the biochemical responses, excluding EROD and MDA, resulted to be analytically detectable in tail tissues of P. siculus, although the mean values obtained with this minimal-invasive method were significantly lower than those obtained with invasive one.