Ecotoxicological and microbiological characterization of soils from heavy-metal- and hydrocarbon-contaminated sites

The aims of this study were to characterize soils from industrial sites by combining physicochemical, microbiological, and ecotoxicological parameters and to assess the suitability of these assays for evaluation of contaminated sites and ecological risk assessment. The soil samples were taken from long-term contaminated sites containing high amounts of heavy metals (sites 1 and 2) or petroleum hydrocarbons (site 3) located in the upper Silesia Industrial Region in southern Poland. Due to soil heterogeneity, large differences between all investigated parameters were measured. Microbiological properties revealed the presence of high numbers of viable hetrotrophic microorganisms. Soil enzyme activities were considerably reduced or could not be detected in contaminated soils. Activities involved in N turnover (N mineralization and nitrification) were significantly (P?<?0.05) higher in samples from the metal-contaminated sites than in samples from the hydrocarbon-contaminated site, whereas the opposite was observed for phosphatase activity. The Microtox test system appeared to be the most appropriate to detect toxicity and significant differences in toxicity between the three sites. The Ostracodtoxkit test was the most appropriate test system to detect toxicity in the hydrocarbon-contaminated soil samples. Correlation analysis between principal components (obtained from factor analysis) determined for physicochemical, microbiological, and ecotoxicological soil properties demonstrated the impact of total and water-extractable contents of heavy metals on toxicity.     

Public health assessment of hexachlorobenzene

Recently, hexachlorobenzene (HCB) was proposed for inclusion in the system of toxicity equivalency factors (TEFs) currently used for dioxin-like compounds. In this paper, we explore the practical implications of the proposition to the Agency for Toxic Substances and Disease Registry (ATSDR) programs by comparing respective health guidance values for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and HCB (expressed as total toxicity equivalents [TEQs]), reviewing possible interactions between HCB and dioxin-like chemicals, and by providing information on actual co-existence of HCB and dioxin-like chemicals at hazardous waste sites. We found a good correlation between the TEF-adjusted oral exposure guidance values for HCB and guidance values for TCDD. The combination of HCB and other dioxin-like compounds was not found in soil, air, or water media at hazardous waste sites. Based on this fact, it is not necessary to include HCB in the total TEQ count at hazardous waste sites at this time.     

The cytotoxic effect of wastewater from the phosphoric gypsum depot on common oak (Quercus robur L.) and shallot (Allium cepa var. ascalonicum)

The effect of wastewater from a phosphoric gypsum depot on common oak, Quercus robur L., at cytogenetical level was studied. Allium-test was used as a control. The treatment of common oak seedlings with wastewater under laboratory conditions caused mitodepressive effect. Chromosome aberrations and mitotic irregularities were found. Cytogenetic analysis of common oak seedlings grown from acorns collected near the depot did not show changes in mitotic activity in comparison to control but the number of aberrations was higher than in control. In comparison to Alliumtest common oak was found to be more tolerant to wastewater from the phosphoric gypsum depot.     

Phenotypic plasticity in Scenedesmus incrassatulus (Chlorophyceae) in response to heavy metals stress

The microalgae genus Scenedesmus is commonly found in freshwater bodies, wastewater facilities and water polluted with heavy metals. Phenotypic plasticity in Scenedesmus has been documented in response to a wide variety of conditions; however, heavy metals have not been comprehensively documented as phenotypic plasticity inducers. In this study, we report the phenotypic plasticity of Scenedesmus incrassatulus (a non-spiny, four-cell coenobium forming species) in response to EC₅₀ value of copper, cadmium and hexavalent chromium. S. incrassatulus was grown in batch cultures in the presence of each metal. Chlorophyll-a content, cell size, parameters derived from the schematic energy-flux model for photosystem II, and morphotype expressions were recorded. Divalent cation metals induced unicellular forms, and hexavalent chromium produced out-of-shape coenobia corresponding to various stages of autospore formation. The changes induced by divalent metals were interpreted as phenotypic plasticity, because they were always associated to population doublings and were reversible when toxicant pressure was removed (only for Cu). Copper was the best inductor of unicellular forms and also affected significantly all the photosynthetic parameters measured. The developed morphotypes could confer ecological advantages to S. incrassatulus in metal stressed environments.     

Genetic Diversity Among Genogroup II Noroviruses and Progressive Emergence of GII.17 in Wastewaters in Italy (2011–2016) Revealed by Next-Generation and Sanger Sequencing

Noroviruses (NoV) are a major cause of gastroenteritis worldwide. Recently, a novel variant of NoV GII.17 (GII.P17_GII.17 NoV), termed Kawasaki 2014, has been increasingly reported in NoV outbreaks in Asia, and has also been described in Europe and North America. In this study, sewage samples were investigated to study the occurrence and genetic diversity of NoV genogroup II (GII) along a 6-year period. Moreover, the spread of GII.17 strains (first appearance and occurrence along time) was specifically assessed. A total of 122 sewage samples collected from 2011 to 2016 from four wastewater treatment plants in Rome (Italy) were initially tested using real-time RT-(q)PCR for GII NoV. Positive samples were subsequently subjected to genotypic characterization by RT-nested PCRs using broad-range primes targeting the region C of the capsid gene of GII NoV, and specific primers targeting the same region of GII.17 NoV. In total, eight different genotypes were detected with the broad-range assay: GII.1 (n = 6), GII.2 (n = 8), GII.3 (n = 3), GII.4 (n = 13), GII.6 (n = 3), GII.7 (n = 2), GII.13 (n = 2), and GII.17 (n = 3), with the latter two genotypes detected only in 2016. Specific amplification of GII.17 NoV was successful in 14 out of 110 positive samples, spanned over the years 2013–2016. The amplicons of the broad-range PCR, pooled per year, were further analyzed by next-generation sequencing (NGS) for a deeper analysis of the genotypes circulating in the study period. NGS confirmed the circulation of GII.17 NoV since 2013 and detected, beyond the eight genotypes identified by Sanger sequencing, three additional genotypes regarded as globally uncommon: GII.5, GII.16, and GII.21. This study provides evidence that GII.17 NoV Kawasaki has been circulating in the Italian population before its appearance and identification in clinical cases, and has become a major genotype in 2016. Our results confirm the usefulness of wastewater surveillance coupled with NGS to study the molecular epidemiology of NoV and to monitor the emergence of NoV strains.     

Occurrence of polybrominated diphenylethers, polychlorinated dibenzo-p-dioxins, dibenzofurans and biphenyls in coastal sediments from Spain

Fifteen sediment samples were analysed in order to determine their content of polybrominated diphenylethers (PBDEs), polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and biphenyls (PCBs). Samples were collected from several hot spots on the Spanish coast, such as the harbours of Almeria and Tarragona, and the mouths of the Besos and Llobregat rivers in Barcelona. A generic analytical procedure based on Soxhlet extraction followed by an automated cleanup system and gas chromatography-ion trap-mass spectrometry was employed for determining the toxic congeners of PCDDs and PCDFs, as well as dioxin-like PCBs. As regards PBDE determinations, a rapid method based on the use of selective pressurized liquid extraction followed by gas chromatography-negative ion chemical ionization-mass spectrometry was applied. Total toxicity equivalent (WHO-TEQ) values were calculated using the toxicity equivalent factors proposed by WHO for dioxin-like PCBs, PCDDs and PCDFs. WHO-TEQ values ranged from 0.3 to 75 pg/g dry weight (dw), with PCB contribution on the toxicity of the samples between 1 and 84%. Total PBDE levels ranged from 2.7 to 134 ng/g dw, with BDE-209 contribution on the total PBDE contamination between 50 and 99%.     

Use of Oxidation Onset Temperature Measurements for Evaluating the Oxidative Degradation of Isotatic Polypropylene

The widespread use of polymeric materials in many applications requires polymers that are weather resistant. The aim of this study is to examine the usefulness of oxidation onset temperature measurements as a tool for evaluating oxidative degradation of isotatic polypropylene. Differential scanning calorimetry was used to measure the oxidation onset temperature in formulations of isotatic polypropylene containing different concentrations of light and thermal stabilizers after aging in a Weather-Ometer. Oxidation onset temperature gave better results than oxidation induction time for evaluating the degradation of isotatic polypropylene during aging in a Weather-Ometer. However, both tests showed a poor correlation with the appearance of cracks seen by optical microscopy.     

Umbilical artery velocity waveforms before and after chorionic villus sampling

Because a vascular aetiology has been suggested for the limb and oromandibular defects described after chorionic villus sampling (CVS), to determine whether transabdominal (TA) CVS causes noticeable changes in umbilical artery velocity waveforms in first-trimester pregnancies, the pulsatility index (PI) of the umbilical artery was evaluated before and after TA-CVS in 175 pregnancies sampled between 10·0 and 13·0 weeks' gestation. In 139 uncomplicated pregnancies, the mean PI values (with 95 per cent confidence interval) were before TA-CVS 2·751 (2·692–2·809), after 10 min 2·723 (2·697–2·809), and after 1 h 2·781 (2·722–2·840). There were no significant changes in PI relative to the CVS procedure either in pregnancies with an abnormal result or in those ending in spontaneous abortion. Our data do not support any statistically significant change in umbilical artery PI relative to TA-CVS in first-trimester pregnancies. This procedure, despite its invasive character, does not appear to affect the feto-placental circulation.     

Effect of different radiotracer labelling techniques on radionuclide excretion from marine organisms

Experiments were designed to assess the effect of different techniques of radiotracer labelling on subsequent radioisotope excretion rates in marine crustaceans. A small amphipod (Gammarus locusta) that accumulated ⁶⁵Zn under a close approximation of natural conditions excreted the radiotracer during a 3-month period at a markedly different rate than those of comparable amphipods labelled with ⁶⁵Zn in the laboratory via different combinations of radioactive food and seawater. Shrimp (Lysmata seticauda) administered ⁶⁵Zn by three different methods in the laboratory displayed different excretion kinetics during the first 2 months of loss. The difference between excretion rates most likely was a reflection of the degree to which the various zinc pools within the shrimp had equilibrated with the radiotracer. During the next several months all ⁶⁵Zn loss rates were quite similar, probably indicating that radiotracer excretion was taking place from similar zinc pools within the shrimp. The importance of adequate radiotracer labelling techniques as well as proper design of subsequent excretion experiments, used to gain reliable information on flux rates of the corresponding stable metal, is discussed. It is concluded that laboratory radiotracer experiments which are intended to supply information on actual situations in the sea require careful design and execution.     

First in vitro use of the phenylphosphate carboxylase enzyme in supercritical CO2 for the selective carboxylation of phenol to 4-hydroxybenzoic acid

A new biotechnological synthetic approach to 4-hydroxybenzoic acid is reported. Thauera aromatica cells produce the phenylphosphate carboxylase enzyme that selectively carboxylates phenylphosphate in the para position, without any formation of the ortho isomer. Here we show that the enzyme also works in supercritical carbon dioxide, opening a new technological solution to the recovery of the products that is a key problem when aqueous media are used. The enzyme shows an activity comparable with that shown under CO₂ pressure in aqueous media and does not require the presence of NaHCO₃.