Carlo Pretti

Acute toxicity of ionic liquids to the zebrafish (Danio rerio)

Acute toxicity and histological damage derived from exposure of Danio rerio (zebrafish) to several ionic liquids have been evaluated.

An optimized digestion method coupled to electrochemical sensor for the determination of Cd, Cu, Pb and Hg in fish by square wave anodic stripping voltammetry

An optimized digestion method coupled to electrochemical detection to monitor lead, copper, cadmium and mercury in fish tissues was developed. Square wave anodic stripping voltammetry (SWASV) coupled to disposable screen-printed electrodes (SPEs) was employed as fast and sensitive electroanalytical method for heavy metals detection. Different approaches in digestion protocols were assessed. The study was focused on Atlantic hake fillets because of their wide diffusion in the human nutrition. Best results were obtained by digesting fish tissue with hydrogen peroxide/hydrochloric acid mixture coupled to solid phase (SP) purification of the digested material. This combined treatment allowed quantitative extraction from fish tissue (muscle) of the target analytes, with fast execution times, high sensitivity and avoiding organic residues eventually affecting electrochemical measurements. Finally, the method has been validated with reference standard materials such as dogfish muscle (DORM-2) and mussel tissues (NIST 2977).

Ecotoxicity of pristine graphene to marine organisms

The ecotoxicity of pristine graphene nanoparticles (GNC1, PGMF) in model marine organisms was investigated. PGMF resulted more toxic than GNC1 to the bioluminescent bacterium Vibrio fischeri and the unicellular alga Dunaliella tertiolecta on the basis of EC50 values (end-points: inhibition of bioluminescence and growth, respectively). No acute toxicity was demonstrated with respect to the crustacean Artemia salina although light microscope images showed the presence of PGMF and GNC1 aggregates into the gut; a 48-h exposure experiment revealed an altered pattern of oxidative stress biomarkers, resulting in a significant increase of catalase activities in both PGMF and GNC1 1mg/L treated A. salina and a significant increase of glutathione peroxidase activities in PGMF (0.1 and 1mg/L) treated A. salina. Increased levels of lipid peroxidation of membranes was also observed in PGMF 1mg/L exposed A. salina.

Interactions between metabolism of trace metals and xenobiotic agonists of the aryl hydrocarbon receptor in the antarctic fish Trematomus bernacchii: Environmental perspectives

Although Antarctica is a pristine environment, organisms are challenged with contaminants either released locally or transported from industrialized regions through atmospheric circulation and marine food webs. Organisms from Terra Nova Bay also are exposed to a natural enrichment of cadmium, but to our knowledge, whether such environmental conditions influence biological responses to anthropogenic pollutants has never been considered. In the present study, the Antarctic rock cod (Trematomus bernacchii) was exposed to model chemicals, including polycyclic aromatic hydrocarbons (benzo[a]pyrene), persistent organic pollutants (2,3,7,8-tetrachlorodibenzo-p-dioxin [TCDD]), cadmium, and a combination of cadmium and TCDD. Analyzed parameters included chemical bioaccumulation, activity, and levels of biotransformation enzymes (cytochrome P4501A); metallothioneins and the efficiency of the antioxidant system measured as individual defenses (catalase, glutathione, glutathione reductase, glutathione S-transferases, and glutathione peroxidases); and total scavenging capacity toward peroxyl and hydroxyl radicals. Reciprocal interactions between metabolism of inorganic and organic pollutants were demonstrated. Dioxin enhanced the accumulation of cadmium, probably stored within proliferating endoplasmic reticulum, and cadmium suppressed the inducibility of cytochrome P4501A, allowing us to hypothesize a posttranscriptional mechanism as the depletion of heme group availability. Clear evidence of oxidative perturbation was provided by the inhibition of antioxidants and enhanced sensitivity to oxyradical toxicity in fish exposed to organic chemicals. Exposure to cadmium revealed counteracting responses of glutathione metabolism; however, these responses did not prevent a certain loss of antioxidant capacity toward peroxyl radicals. The pattern of antioxidant responses exhibited by fish coexposed to cadmium and TCDD was more similar to that observed for cadmium than to that observed for TCDD. The overall results suggest that elevated natural levels of cadmium in Antarctic organisms from Terra Nova Bay can limit biotransformation capability of polycyclic (halogenated) hydrocarbons, thus influencing the bioaccumulation and biological effects of these chemicals in key sentinel species.

Biotransformation enzymes and their induction by β-naphtoflavone in adult sea bass (Dicentrarchus labrax)

Abstract In the present study we have determined for the first time the hepatic activities of several microsomal P-450 dependent monooxygenases including the testosterone hydroxylase and the Ah-receptor gene battery-related phase II enzymes [UDP-glucuronyl transferase (UDP-GT), DT-diaphorase, glutathione S -transferase, aldehyde dehydrogenases] in adult sea bass (300–400 g). We also investigated the effect of β -naphtoflavone ( β -NF) on these enzymes to assess their potential as biomarkers of polycyclic aromatic hydrocarbon exposure. β -NF, administered to fish with a single i.p. injection at various doses, resulted in an induction in the liver of the P-450 1A, as immunodetected by anti rat P-450 1A1 antibodies, P-450 1A-linked ethoxyresorufin- O -deethylase (EROD) and methoxyresorufin- O -demethylase (MEROD) activities. Among the phase II enzymes, only the phenol UDP-GT was induced in liver by β -NF. The time course of these inductions exhibited different patterns. EROD and MEROD activities reached a peak value 3 days post-injection and maintained values significantly above the corresponding control activities over the 30 days of the experiment, the phenol UDP-GT activity peaked at day 8 and afterwards declined rapidly achieving the control value 18 days post-injection. A treatment of 80 mg kg −1 β -NF increased the immunodetectable P-450 1A apoprotein and EROD, but not MEROD, activity in extrahepatic tissues such as, gill, kidney and olfactory rosettes. Taken overall, the results indicate that MEROD, EROD and phenol UDP-GT activities may be useful biomarkers for the exposure of adult sea bass to polycyclic aromatic hydrocarbons.

Antimicrobial susceptibility of Staphylococcus intermedius and Staphylococcus schleiferi isolated from dogs.

The susceptibility to 23 antimicrobial agents was determined in 114 isolates of Staphylococcus intermedius and eight isolates of Staphylococcus schleiferi of canine origin. Overall, 73% of S. intermedius isolates and 37.5% of S. schleiferi isolates were susceptible to all the 23 antimicrobials tested. The large majority of S. intermedius strains retained susceptibility to antimicrobials currently employed in treatment of pyoderma (cephalosporins, cotrimoxazole and association amoxicillin-clavulanic acid) as well as to those effective against staphylococci (fusidic acid, rifampicin and fluoroquinolones). Resistance in S. intermedius was observed mainly against macrolides, chloramphenicol and lincosamides, while S. schleiferi isolates retained susceptibility to all antimicrobials except three of six fluoroquinolones. Although, our results confirm susceptibility to antimicrobials currently employed in pyoderma treatment, the several different resistance patterns observed for S. intermedius emphasize the importance of antimicrobial susceptibility testing of canine staphylococci to choose the most appropriate treatment of infections and to allow the prudent use of antimicrobial drugs in companion animals.

Effects of β-naphthoflavone on the cytochrome P450 system, and phase II enzymes in gilthead seabream (Sparus aurata)

Abstract The effect of β-naphthoflavone (β-NF) on several catalytic activities of cytochrome P450 (CYP) and phase II enzymes putatively controlled by [Ah]-receptor activation in the liver, heart and kidney of gilthead seabream, was investigated. In the liver, β-NF treatment [intraperitoneal injection (i.p.) 50 mg/kg] resulted in an increase of CYP content, immunoreactive CYP 1A and methoxyresorufin-O-demethylase (MEROD), pentoxyresorufin O-depentylase (PROD) and ethoxyresorufin-O-deethylase (EROD) activities. However, β-NF had no effect on any of the hepatic phase II enzymes examined (benzaldehyde dehydrogenase, propionaldehyde dehydrogenase, glutathione S-transferase, UDP-glucuronyl-transferase, DT-diaphorase). Single i.p. injection of 10 mg/kg β-NF showed a maximal induction of CYP 1A-like protein and EROD activity after 3–7 days. CYP 1A and EROD returned to control levels 18-days post-treatment. β-NF injection also caused a rapid increase of a single band size of mRNA recognized by a CYP 1A1 cDNA fragment from sea bass (Dicentrarchus labrax). Expression of mRNA preceded the increase of EROD activity and declined rapidly by 96 h. Dose–response experiments demonstrated that EROD was significantly enhanced in liver by a single injection of 0.3 mg/kg β-NF and was the most sensitive measurement for CYP 1A-like induction. β-NF treatments also increased the expression of CYP 1A-like protein, mRNA and EROD, but not MEROD and PROD activities in heart and kidney.

Photopolymerized network polysiloxane films with dangling hydrophilic/hydrophobic chains for the biofouling release of invasive marine serpulid Ficopomatus enigmaticus

Novel photopolymerized network films based on a polysiloxane matrix containing varied amounts of polyoxyethylene (P3) or perfluorohexylethyl (F) dangling side chains were investigated. For films containing less than 10 wt % P3 and F, the wettability and elastic modulus were similar to those of the photopolymerized network matrix. However, angle-resolved X-ray photoelectron spectroscopy measurements proved that the surface of films with F dangling chains was highly enriched in fluorine depending on both the amount of P3 and F and their relative ratio in the films. The biological performance of the films was evaluated against a new widespread and invasive marine biofoulant, the serpulid Ficopomatus enigmaticus. The diatom Navicula salinicola was also assayed as a conventional model organism for comparison. Films richer in P3 better resisted the settlement and promoted the release of calcified tubeworms of F. enigmaticus.

The impacts of emergent pollutants on Ruditapes philippinarum : biochemical responses to carbon nanoparticles exposure

Multi-walled carbon nanotubes (MWCNTs) are one of the most important carbon Nanoparticles (NPs). The production and use of these NPs are increasing rapidly and, therefore, the need to assess their presence in the environment and associated risks has become of prime importance. Recent studies demonstrated the impacts of different NPs on bivalves, a taxonomic group where species tolerance to anthropogenic stressors, such as pollutants, is widely variable. The Manila clam Ruditapes philippinarum is one of the most commonly used bivalve species in environmental monitoring studies and ecotoxicology tests, however, to our knowledge, no information is available on biochemical alterations on this species due to MWCNTs exposure. Thus, the present study aimed to assess the toxic effects of different MWCNT concentrations (0.01; 0.10 and 1.00mg/L) in R. philippinarum biochemical (energy reserves, metabolic capacity, oxidative status and neurotoxicity) performance, after 28days of exposure. The results obtained revealed that exposure to MWCNTs altered energy-related responses, with higher metabolic capacity and lower glycogen and protein concentrations in clams exposed to these carbon NPs. Moreover, R. philippinarum exposed to MWCNTs showed oxidative stress expressed in higher lipid peroxidation and lower ratio between reduced and oxidized glutathione, despite the activation of defence mechanisms in exposed clams. Additionally, neurotoxicity was observed by inhibition of cholinesterases activity in organisms exposed to MWCNTs. The present study provides valuable information regarding how these emerging pollutans could become a potential risk for the environment and living organisms.

An optical immunosensor for rapid vitellogenin detection in plasma from carp (Cyprinus carpio)

Vitellogenin (vtg) has proven to be a sensitive and simple biomarker for assessing exposure of fish to environmental estrogens. The aim of this work was to develop a rapid, in the order of minutes, screening method for the detection of fish vtg. The surface plasmon resonance technique (Biacore Xtrade mark) was coupled with immunodetection for the determination of fish vtg in plasma and mucus from carp (Cyprinus carpio). Monoclonal anti-vtg antibodies were linked on the sensor surface through chemical cross-linking via a capturing antibody. A simple regeneration process allowed the reuse of the sensor surface. Sensor optimisation was carried out using carp vtg. The developed immunosensor was tested with vtg spiked samples and with plasma and mucus from fish exposed to 17beta-estradiol (E2). Vitellogenin could be detected in the ppm range in buffer as well as in plasma and mucus. Good discrimination between control and exposed samples was obtained. The results were compared with ELISA and a correlation coefficient of R(2)=0.85 (n=9) between the two methods indicated that the immunochemical biosensor could be used for the analysis of vtg in fish plasma samples. The assay time was 20min hence allowing for rapid sample screening.