Contamination Levels and Preliminary Assessment of the Technical Feasibility of Employing Natural Attenuation in 5 Priority Areas of Presidente Bernardes Refinery in Cubatão, São Paulo, Brazil

Five priority areas of potential impact by contaminants (API) were investigated at the Presidente Bernardes Refinery in Cubatão, São Paulo, Brazil with the following aims: (i) to identify both organic and inorganic contaminants present in soil and groundwater; (ii) to define the environmental conditions relevant for microbial activity at the site and (iii) to evaluate the feasibility of employing natural attenuation for treatment of the hydrocarbon contamination. One area (API 1) was an uncontrolled landfill, where waste materials from the refinery were deposited between 1954 and 1986, and four areas (API 4, 5, 7 and 11) were located in the operational section of the refinery. Soil contamination by regulated BTEX compounds (benzene, toluene, total xylenes) was restricted to two samples from API 1. Nonregulated ethylbenzene was detected in one soil sample from API 4, one from API 5 and two from API 1. No soil contained regulated PAH above threshold levels. Several nonregulated PAHs were found in 6 soil samples from API 1, 3 soil samples from API 4 and 1 soil sample from API 5. Site soils contained very high aluminium concentrations, but metal contamination was restricted to one soil sample from API 1, which contained nickel above threshold limits. BTEX contamination of groundwater was due mostly to benzene. Of the 17 PAH molecules tested, only naphthalene and 2-methylnaphthalene occurred in groundwater. The sum of total BTEX and total PAH exceeded 200 μg/L in only a few monitoring wells in API 4, 5 and 11 and was always below 2.640 μg/L. Be, Cd, Cr, Cu, Hg, Ni, Se, Ag, Tl and Zn were not detected in groundwater, which was contaminated in a few locations by aluminium (mostly below 1 mg/L), lead (<0.066 mg/L) and arsenic (<0.056 mg/L). S, K, Ca, Mg and Fe were present in groundwater in excess of physiological requirements for microbial growth, but low concentrations of N and P could become growth limiting. However, BTEX were efficiently degraded in saturated and unsaturated zone microcosms and nutrient amendments did not stimulate biodegradation rates measurably. The inorganic carbon pool in groundwater was up to one order of magnitude larger than the organic carbon pool. Total inorganic carbon (TIC) in API groundwater exceeded TIC of clean groundwater by factors of 2 (API 4), 6 (API 5, 7 and 11) or 10 (API 1). Most of the inorganic carbon incorporated into groundwater beneath the refinery originated from biodegradation in the unsaturated soil, which contained a microbiota (10⁶ cells/g on average) capable of growth with most of the pure (benzene, toluene, ethylbenzene and xylene) and mixed hydrocarbons tested (diesel oil, gasoline, naphtha, condensate, aromatic residue and fuel oil). A viscous hydrocarbon paste uncovered in API 1 was insoluble in water but dissolved in dichloromethane. Many organic components of this paste were biodegradable as evidenced by weight reduction of the hydrocarbon paste and by the growth of suspended and attached biomass in saturated zone microcosms, where the paste was the only carbon source. This study indicates that monitored natural attenuation may be a technically feasible and efficient means for plume control in API 1, 4 and 5, provided the plumes in API 4 and 5 are not expanding. This technique is not suitable for contaminant reduction in API 11.     

Leaching composted lignocellulosic wastes to prepare container media: Feasibility and environmental concerns

The leaching of salt and mineral elements from three composts prepared with residual vegetable crop biomass (melon, pepper or zucchini) was studied using methacrylate columns and distilled water. The benefits of the leached composts to be used for ornamental potted plant production were also analysed. After leaching 5 container capacities of effluent, both the electrical conductivity and the concentration of soluble mineral elements in compost leachates decreased substantially and remained close to the target levels. Composts reacted differently to leaching due to differences in the raw waste sources and the composting process and hence, in their physical and chemical characteristics. At the end of the experiment, after pouring 8 container capacities of water, the leaching efficiency of the salts was 96%, 93% and 87% for melon, pepper and zucchini-based composts, respectively. Mineral elements differed in their ability to be removed from the composts; N (NH₄⁺ and NO₃^?), K⁺, Na⁺, Cl^?, and SO₄^2? were leached readily, whereas H₂PO₄^?, Ca²⁺, and Mg²⁺ were removed hardly. Leached composts showed a range of physico-chemical and chemical characteristics suitable for use as growing media constituents. Potted Calendula and Calceolaria plants grew in the substrates prepared with the leached composts better than in those made with the non-leached ones. Finally, special emphasis must be paid to the management of the effluents produced under commercial conditions to avoid environmental pollution.     

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.     

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.     

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.     

Phylogeography of California and Galápagos sea lions and population structure within the California sea lion

We investigate the phylogeography of California (Zalophus californianus) and Galápagos (Z. wollebaeki) sea lions and describe within-population structure for the California sea lion based on mitochondrial DNA. Fifty control-region haplotypes were found, 41 from Z. californianus and 9 from Z. wollebaeki, with three fixed differences between the two species. Ranked population boundaries along the range of Z. californianus were defined based on the Monmonier Maximum Difference Algorithm, resulting in five genetically distinct populations, two in the Pacific Ocean and three inside the Gulf of California. A Minimum Spanning Network showed a strong phylogeographic signal with two well-defined clusters, Z. californianus and Z. wollebaeki, separated by six base-pair differences, supporting the existence of two genetically distinct species with an estimated divergence time of ~0.8 Ma. Results are discussed in the context of the historical geologic and paleoceanographic events of the last 1 Ma in the eastern Pacific.     

Embryogenesis of decapod crustaceans with different life history traits,feeding ecologies and habitats: a fatty acid approach

Variations in embryo size and fatty acid (FA) dynamics during embryogenesis were evaluated in deep-sea pandalids and portunid swimming crabs from the Portuguese continental margin and Madeira Island slope and compared with previous data on neritic and deep-sea lobsters and shrimps (collected between February 2001 and March 2004). Inter-specific variations in embryo size seem to be dictated primarily by phylogeny rather than by differences in reproductive or early life history traits. FA reserves were significantly correlated with embryo size (P < 0.001). Principal component analysis revealed differences among three groups (1—neritic caridean shrimps, 2—deep-sea pandalids of the genus Plesionika, and lobsters, 3—portunid crabs and the deep-sea pandalid Chlorotocus crassicornis, Costa 1871). Group 1 was clearly separated by PC1 mainly due to the higher percentage of essential C₁₈ (linoleic and linolenic acids) and C₂₀ (namely eicosapentaenoic) polyunsaturated FA (specific markers of primary producers). PC2 separated Group 2 from Group 3 due to differences in the percentage of several saturated FA (including odd-numbered FA—bacterial markers) and C₁₈ monounsaturated FA (namely 18:1n − 9, a general marker of carnivory). Therefore, these differences among groups seem to result from distinctions in diet and ecological niche. Intra-specific differences in FA composition between western and southern Plesionika martia martia (A. Milne-Edwards, 1883) populations may reflect higher water temperatures on the south sub-tropical coast. Lobster embryonic development was more demanding of lipid energy than that of the other decapod species, which may reflect an evolutionary trend in decapod taxa related to an increasing degree of lecithotrophy. However, a lower FA catabolism can be interpreted as an enhanced independence of the newly hatched larvae from external energy sources. Higher FA content at hatching and, as a consequence, a greater independence from the external environment should increase the chances of larval survival. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Assessing the effects of pseudo-absences on predictive distribution model performance

Modelling species distributions with presence data from atlases, museum collections and databases is challenging. In this paper, we compare seven procedures to generate pseudo-absence data, which in turn are used to generate GLM-logistic regressed models when reliable absence data are not available. We use pseudo-absences selected randomly or by means of presence-only methods (ENFA and MDE) to model the distribution of a threatened endemic Iberian moth species (Graellsia isabelae). The results show that the pseudo-absence selection method greatly influences the percentage of explained variability, the scores of the accuracy measures and, most importantly, the degree of constraint in the distribution estimated. As we extract pseudo-absences from environmental regions further from the optimum established by presence data, the models generated obtain better accuracy scores, and over-prediction increases. When variables other than environmental ones influence the distribution of the species (i.e., non-equilibrium state) and precise information on absences is non-existent, the random selection of pseudo-absences or their selection from environmental localities similar to those of species presence data generates the most constrained predictive distribution maps, because pseudo-absences can be located within environmentally suitable areas. This study shows that if we do not have reliable absence data, the method of pseudo-absence selection strongly conditions the obtained model, generating different model predictions in the gradient between potential and realized distributions.     

Determination of glyoxal and methylglyoxal in atmospheric particulate matter by 2,4-dinitrophenylhydrazine derivatisation

A simple and rapid method has been optimised to determine glyoxal and methylglyoxal in atmospheric particulate matter by extraction and derivatisation with 2,4-dinitrophenylhydrazine solutions. Minimal sample preparation was required by mixing a portion of filter charged with atmospheric particulate matter with 5?mL of a saturated solution of 2,4-dinitrophenylhydrazine (1.6?g?L^?1) in acetonitrile and acidified by H₂SO₄ (50?µL?L^?1). Chromatographic analyses were carried out 1?h after derivatisation solution was mixed. Finally, the proposed method was applied to quantify these dicarbonyls in atmospheric particulate matter samples, which were obtained as part of a large field study to characterise atmospheric aerosol from a rural area of Madrid. Glyoxal concentrations averaged 0.24?ng?m^?3, whereas mean values of methyl glyoxal were around 0.95?ng?m^?3. Methyl glyoxal levels showed a clear relationship with the particulate matter levels, whereas glyoxal data did not show such a correlation. This could indicate that both dicarbonyls come from different sources.