Alfredo Parrella

Toxicity identification evaluation of leachates from municipal solid waste landfills: a multispecies approach

The toxicity of leachates from two municipal solid waste (MSW) landfills in Southern Italy was characterized using a toxicity identification evaluation procedure. The chemical and physical fractionation techniques were: pH adjustment, pH adjustment/filtration, pH adjustment/C(18) solid phase extraction, graduated pH and EDTA chelation. All the samples exhibited acute toxicity towards the bacterium Vibrio fischeri, the freshwater rotifer Brachionus calyciflorus and the freshwater crustaceans Thamnocephalus platyurus and Daphnia magna. Statistical techniques were used to determine the discriminatory power and the toxicity detection capacity of the different assays and to choose a minimal battery of tests for the toxicity identification of leachates. Toxicity was closely associated with pH, generally increasing at higher pH levels and decliming at lower ones. Furthermore, results showed that toxicants could be characterized as cations, basic chemicals, suspended solids and apolar compounds.

In situ monitoring of urban air in Southern Italy with the tradescantia micronucleus bioassay and semipermeable membrane devices (SPMDs)

This study was designed to assess toxic and genotoxic compounds in the urban air of Caserta, South Italy using cuttings from the plant Tradescantia #4430. In situ monitoring of gaseous pollutans was made at 17 sampling points in two seasons of the year. Genotoxicity was evaluated by recording the micronuclei in meiotic pollen mother cells (Trad-MCN assay). In addition, the passive sampler semipermeable membrane devices (SPMDs) were deployed at the sampling points with a significant increase in micronuclei frequency. SPMDs concentrated priority organic pollutants were identified by high performance liquid chromatography and gas chromatography, while toxicity and mutagenesis were assessed on the bacterium Vibrio fischeri using the Microtox and Mutatox systems respectively. Significant toxic and mutagenic effects were observed at different points on the town grid and SPMDs effectively concentrated trace contaminants. The relationship between what was present in the air sampled by SPMDs and the micronuclei frequency was also explored.

Acute and chronic toxicity of six anticancer drugs on rotifers and crustaceans.

The growing use of cytostatic drugs is gaining relevance as an environmental concern. Environmental and distribution studies are increasing due to the development of accurate analytical methods, whereas ecotoxicological studies are still lacking. The aim of the present study was to investigate the acute and chronic toxicity of six cytostatics (5-fluorouracil, capecitabine, cisplatin, doxorubicin, etoposide, and imatinib) belonging to five classes of Anatomical Therapeutic Classification (ATC) on primary consumers of the aquatic chain (Daphnia magna, Ceriodaphnia dubia, Brachionus calyciflorus, and Thamnocephalus platyurus). Acute ecotoxicological effects occurred at concentrations in the order of mgL(-)(1), higher than those predicted in the environment, and the most acutely toxic drugs among those tested were cisplatin and doxorubicin for most aquatic organisms. For chronic toxicity, cisplatin and 5-fluorouracil showed the highest toxic potential in all test organisms, inducing 50% reproduction inhibition in crustaceans at concentrations on the order of μgL(-)(1). Rotifers were less susceptible to these pharmaceuticals. On the basis of chronic results, the low effective concentrations suggest a potential environmental risk of cytostatics. Thus, this study could be an important starting point for establishing the real environmental impact of these substances.

Estrogenic activity of pharmaceuticals in the aquatic environment.

In the last years pharmaceuticals have aroused great interest as environmental pollutants for their toxic effects towards non target organisms. This study wants to draw attention to a further adverse effect of drugs, the endocrine interference. The most representative drugs of the widespread classes in environment were investigated. The YES-test and the E-screen assay were performed to detect the capability of these substances to bind the human estrogenic receptor alpha (hER alpha) in comparison with 17beta-estradiol. Out of 14 tested pharmaceuticals, 9 were positive to YES-assay and 11 were positive to E-screen assay; in particular, Furosemide and the fibrates (Bezafibrate, Fenofibrate and Gemfibrozil) gave the maximal estrogenic response. Tamoxifen showed its dual activity as agonist and antagonist of hER alpha.

Effects of ranitidine and its photoderivatives in the aquatic environment.

This study was designed to assess the overall ecotoxicity of ranitidine, a histamine H(2)-receptor antagonist that inhibits stomach acid production. Hence, in addition to ranitidine, its main two photoderivatives, obtained by solar simulator irradiation in water, were investigated. The photoproducts were identified by their physical features. Bioassays were performed on rotifers and microcrustaceans to assess acute and chronic toxicity, while SOS Chromotest and Ames test were utilized to detect the genotoxic potential of the investigated compounds. The results showed that ranitidine did not show any acute toxicity at the highest concentration tested (100 mg/L) for all the organisms utilized in the bioassays. Chronic exposure to these compounds caused inhibition of growth population on rotifers and crustaceans. Genotoxic and mutagenic effects were especially found for one photoproduct suggesting that transformation products, as frequently demonstrated, may show effects higher than the respective parental compound.

Eco-genotoxicity of six anticancer drugs using comet assay in daphnids.

The eco-genotoxicity of six anti-neoplastic drugs, 5-fluorouracil, capecitabine, cisplatin, doxorubicin, etoposide, and imatinib, belonging to five classes of anatomical therapeutic classification (ATC), was studied applying the in vivo comet assay on cells from whole organisms of Daphnia magna and Ceriodaphnia dubia. For the first time, this test was performed in C. dubia. In addition, to have a wider genotoxic/mutagenic profile of the anticancer drugs selected, SOS chromotest and Salmonella mutagenicity assay were performed. The comet results showed that all drugs induced DNA damage, in both Cladocerans, with environmental concern; indeed Doxorubicin induced DNA damage in the order of tens of ng L(-1) in both crustaceans, as well as 5-flurouracil in C. dubia and cisplatin in D. magna. In the SOS Chromotest all drugs, except imatinib, were able to activate the repair system in Escherichia coli PQ37 while in the Salmonella mutagenicity assay, doxorubicin was the only drug able to cause direct and indirect frameshift and base-pair substitution mutations. Comet assay was the most sensitive tool of genotoxic exposure assessment, able to detect in vivo the adverse effects at concentration lower than those evaluated in vitro by bacterial assays.

Estrogenic activity and cytotoxicity of six anticancer drugs detected in water systems

The aim of the present study was to investigate the in vitro estrogenic and the cytotoxic activity of six cytostatics (5-fluorouracil, capecitabine, cisplatin, doxorubicin, etoposide, and imatinib) belonging to the five classes of Anatomical Therapeutic Classification (ATC) detected in wastewater systems. The estrogenic activity was assessed by YES-assay on Saccharomyces cerevisiae-RMY326 and E-screen on MCF-7 cells. The cytotoxic activity was assessed by MTT Cell Proliferation Assay on the MCF-7 and the MDA-MB-231 cells. The results of estrogenic activity, detected by E-screen and expressed as EC50, showed a high potential of imatinib (10(-7) μM) followed by cisplatin and 5-fluorouracil. Capecitabine was poorly estrogenic while etoposide and doxorubicin EC50 values were not possible to determine. Cytotoxicity was found at concentrations far from those detected in effluents. The potential endocrine activity of the most active drugs could be associated with human and wildlife risk when considering their occurrence in the environment.

Toxicity of exposure to binary mixtures of four anti-neoplastic drugs in Daphnia magna and Ceriodaphnia dubia.

Anticancer drugs, interfering with DNA in every living organism, may pose a threat to aquatic environment, even more when they occur as complex mixtures. We investigated the combined long term toxic potential of four anti-neoplastic drugs (5-fluorouracil [5-FU], cisplatin [CDDP], etoposide [ET] and imatinib mesylate [IM]) testing their binary mixtures on two primary consumers of the freshwater aquatic chain with close phylogenetic relationship: Daphnia magna and Ceriodaphnia dubia. The combined toxicities were assessed using two distinct effect sizes that should be observed if Bliss independence holds. Direct statistical comparison by analysis of variance of single and combined toxicities under the assumption of Bliss independence allowed to accept or reject the independency hypothesis. Independency was confirmed for all mixtures both in D. magna and in C. dubia, except for IM+ ET and IM+CDDP in D. magna and for ET+CDDP and ET+5-FU in C. dubia which at the highest concentrations showed an antagonistic interaction. A synergic tendency was found testing IM+CDDP on C. dubia at the lowest concentration selected. Thus, the chronic ecotoxicological data evaluated in this study show not only a potential environmental risk of anticancer drugs, especially considering their potential synergistic effects, but also the necessity to integrate statistical models with experimental data to establish the real environmental impact of such compounds.

Influence of alkylphenols and trace elements in toxic, genotoxic, and endocrine disruption activity of wastewater treatment plants

Toxicity and endocrine interference of influent and effluent waters from domestic and industrial wastewater treatment plants were determined. In addition, chemical analyses were performed to detect the presence of 17beta-estradiol, 17alpha-ethinyl estradiol, nonylphenol, 4-octylphenol, and p-t-octylphenol as well as lead, copper, and cadmium in these matrices. The results showed that despite low acute toxic potential, most of the samples tested showed both genotoxicity and endocrine interference. Furthermore, to establish whether the observed effects were caused by the alkylphenols and the heavy metals detected, toxic, genotoxic, and endocrine interference tests also were performed on pure chemicals. The acute toxicity was measured on the crustacean Daphnia magna. The estrogenic activity was determined by using the yeast estrogen screen with Saccharomyces cerevisiae RMY326, whereas the SOS Chromotest and Ames test detected the genotoxicity on Escherichia coli PQ37 and Salmonella typhimurium TA98 and TA100, respectively. The results showed that the toxicity found in the matrices did not match the values found for pure chemicals, but a clear correlation was found between alkylphenols and genotoxicity. Both heavy metals and alkylphenols took part in the endocrine interference activity.

Genotoxicity of aqueous extract from heated cooking oils and its suppression by lactobacilli.

In the present study the mutagenic and genotoxic effects of aqueous extracts from six cooking oils (extra vergine olive, peanut, sunflower, soybean, corn, and various seeds oils) heated to the respective smoke point were investigated. The Ames test and the SOS Chromotest were carried out for this evaluation. The same oils were also tested after their re-frying, simulating domestic reuse process. Furthermore, the ability of different lactobacilli to reduce the potential genotoxic activity of the fried and re-fried oils was determined applying SOS Chromotest after co-incubation of samples with lactobacilli. The results showed that all the fried oils did not produce mutagenic effects while they induced a SOS response with the highest induction factor for the corn oil. Double heat-treatment caused an increase of the genotoxic activity until two times the first heating. The most susceptible oil to the re-frying procedure was the sunflower oil. The antigenotoxicity results were expressed as percent of genotoxicity inhibition. All the tested strains of lactobacilli exhibited antigenotoxic properties on the fried oils.