Phytoremedial assessment of flora tolerant to heavy metals in the contaminated soils of an abandoned Pb mine in Central Portugal

Significant accumulation of heavy metals in soils and flora exists around the abandoned Barbadalhos Pb mine in Central Portugal. Soil and plant samples [49 species] were collected from two line transects, LT 1 and LT 2, in the mineralized and non-mineralized area, respectively to gain a comprehensive picture of heavy metals in soils and flora to assess its potential for phytoremediation. Phytosociological inventories of the vegetation were made using the Braun-Blanquet cover-abundance scale. Metal concentrations in soil ranged from (in mg kg^?1): 98–9330 [Pb], 110–517 [Zn], 7.1–50 [Co], 69–123 [Cr], 31–193 [Cu], 33 400–98 500 [Fe], 7.7–51 [Ni], 0.95–13 [Ag], 2.8–208 [As], and 71–2220 [Mn] along LT 1; and 24–93 [Pb], 30–162 [Zn], 3.7–34 [Co], 61–196 [Cr], 21–46 [Cu], 24 100–59 400 [Fe], 17–87 [Ni], 0.71–1.9 [Ag], 4.3–12 [As], and 44–1800 [Mn] along LT 2. Plant metal content ranged from (in mg kg^?1): 1.11–548 [Pb], 7.06–1020 [Zn], 0.08–2.09 [Co], 0.09–2.03 [Cr], 2.63–38.5 [Cu], 10.4–4450 [Fe], 0.38–8.9 [Ni], and 0.03–1.9 [Ag] along LT 1; and 0.94–11.58 [Pb], 2.83–96.5 [Zn], 0.12–1.44 [Co], 0.21–1.49 [Cr], 1.61–22.7 [Cu], 4.6–2050 [Fe], 0.51–4.81 [Ni], and 0.02–0.31 [Ag] along LT 2. Plants with highest uptake of metals were: Cistus salvifolius (548 mg Pb kg^?1), Digitalis purpurea (1017 mg Zn kg^?1 and 4450 mg Fe kg^?1). Mentha suavolens and Ruscus ulmifolius were seen to hyperaccumulate Ag (1.9 and 1 mg Ag kg^?1, respectively). More metals and higher concentrations were traced in plants from LT 1, especially for Pb and Zn.     

Cytogenetic biomonitoring of primary school children exposed to air pollutants: micronuclei analysis of buccal epithelial cells

There is an increasing attempt in the world to determine the exposures of children to environmental chemicals. To analyze the genotoxic effect of air pollution, micronucleus (MN) assay was carried out in buccal epithelial cells (BECs) of children living in an urban city of Turkey. Children from two schools at urban-traffic and suburban sites were investigated in summer and winter seasons for the determination of BEC-MN frequency (per mille) and frequency of BEC with MN (per mille). The same children were also recruited for lung function measurements within a MATRA project (“Together Towards Clean Air in Eskisehir and Iskenderun”) Measured NO₂ and SO₂ concentrations did not exceed the European Union (EU) limit levels either in urban-traffic or suburban regions. Higher O₃ concentrations were measured in the suburban site especially in the summer period. Particulate matter (PM_2.5 and PM₁₀) levels which did not differ statistically between two regions were above the EU limits in general. Although BEC-MN frequencies of children living in the suburban sites were higher in general, the difference between two regions was not significant either in the summer or winter periods. BEC-MN frequencies of the urban-traffic children were found to be significantly higher in summer period (mean ± SD, 2.68?±?1.99) when compared to winter period (1.64?±?1.59; p?=?0.004). On the other hand, no seasonality was observed for the suburban children. Similar results have been obtained in the BEC frequency with MN in our study. In summer, BEC-MN frequencies were significantly increased with the decrease in pulmonary function levels based on forced expiratory flow between 25 and 75 % of vital capacity (FEF_25–75 %) levels (p?<?0.05). As a conclusion, children living in urban-traffic and suburban areas in the city of Eski?ehir exhibited similar genotoxicity. Seasonal variation in genotoxicity may be interpreted as relatively high ozone levels and increasing time spent at outdoors in the summer.     

Assessment of Jatropha curcas L. biodiesel seed cake toxicity using the zebrafish (Danio rerio) embryo toxicity (ZFET) test

Consequent to the growing demand for alternative sources of energy, the seeds from Jatropha curcas remain to be the favorite for biodiesel production. However, a significant volume of the residual organic mass (seed cake) is produced during the extraction process, which raises concerns on safe waste disposal. In the present study, we assessed the toxicity of J. curcas seed cake using the zebrafish (Danio rerio) embryotoxicity test. Within 1-h post-fertilization (hpf), the fertilized eggs were exposed to five mass concentrations of J. curcas seed cake and were followed through 24, 48, and 72 hpf. Toxicity was evaluated based on lethal endpoints induced on zebrafish embryos namely egg coagulation, non-formation of somites, and non-detachment of tail. The lowest concentration tested, 1 g/L, was not able to elicit toxicity on embryos whereas 100 % mortality (based also on lethal endpoints) was recorded at the highest concentration at 2.15 g/L. The computed LC50 for the J. curcas seed cake was 1.61 g/L. No further increase in mortality was observed in the succeeding time points (48 and 72 hpf) indicating that J. curcas seed cake exerted acute toxicity on zebrafish embryos. Sublethal endpoints (yolk sac and pericardial edema) were noted at 72 hpf in zebrafish embryos exposed to higher concentrations. The observed lethal endpoints induced on zebrafish embryos were discussed in relation to the active principles, notably, phorbol esters that have remained in the seed cake even after extraction.     

Genotoxicity and histological alterations in grey mullet Mugil liza exposed to petroleum water-soluble fraction (PWSF)

Petroleum hydrocarbons are considered one of the main organic chemicals found in water bodies. In the present study, the median lethal concentration (LC₅₀) was estimated for mullet Mugil liza after acute exposure to petroleum water-soluble fraction (PWSF). Furthermore, histopathological studies and micronuclei frequency were also performed in order to observe deleterious effects of medium-term exposition to PWSF. Mullets (25?±?2.3 g) were exposed to chronic concentrations (1.7, 3.5 and 7 % of PWSF), plus the control group, for 14 and 7 days of clearance time. Throughout the experimental period (1, 4, 14 and 21 days), blood samples were collected for analysis of micronucleus (MN) and liver and gills for histopathological study. For these procedures, seven fish were sampled per concentration tested. The LC₅₀-96 h was estimated at 37.5 % of the PWSF. The time required for MN induction was 96 h of exposure. The time of clearance was sufficient to achieve a MN frequency similar to that of the control group. Histopathological studies showed severe changes in the gill and liver tissues. The most relevant histopathology in the gills was telangiectasia. Hepatic histopathology such as cholestasis, dilated sinusoids and inflammatory infiltrates were commonly described. The MN test and histological study effectively detected damages caused by medium-term exposition to the PWSF, and despite the toxicity, a few days without exposure can minimize PWSF genotoxicity in juveniles of M. liza.     

Cyprodinil retention on mixtures of soil and solid wastes from wineries. Effects of waste dose and ageing

In spite of its wide-world economic relevance, wine production generates a huge amount of waste that threatens the environment. A batch experiment was designed to assess the effect of the amendment of an agricultural soil with two winery wastes (perlite and bentonite wastes) in the immobilization of cyprodinil. Waste addition (0, 10, 20, 40, and 80 Mg ha^?1) and different times of incubation of soil-waste mixtures (1, 30, and 120 days) were tested. The addition of wastes improved the soil’s ability to immobilize cyprodinil, which was significantly correlated to total C content in soil-waste mixtures. Longer incubation times decreased the cyprodinil sorption possibly due to the mineralization of organic matter but also as a consequence of the high pH values reached after bentonite waste addition (up to 10.0). Cyprodinil desorption increased as the amount of waste added to soil, and the incubation time increased. The use of these winery wastes contributes to a more sustainable agriculture preventing fungicide mobilization to groundwater.     

Detection of Listeria monocytogenes in ready-to-eat food by Step One real-time polymerase chain reaction

The aim of this study was to follow contamination of ready-to-eat food with Listeria monocytogenes by using the Step One real time polymerase chain reaction (PCR). We used the PrepSEQ Rapid Spin Sample Preparation Kit for isolation of DNA and MicroSEQ® Listeria monocytogenes Detection Kit for the real-time PCR performance. In 30 samples of ready-to-eat milk and meat products without incubation we detected strains of Listeria monocytogenes in five samples (swabs). Internal positive control (IPC) was positive in all samples. Our results indicated that the real-time PCR assay developed in this study could sensitively detect Listeria monocytogenes in ready-to-eat food without incubation.     

Mycobiota and mycotoxins in bee pollen collected from different areas of Slovakia

Contamination by microscopic fungi and mycotoxins in different bee pollen samples, which were stored under three different ways of storing as freezing, drying and UV radiation, was investigated. During spring 2009, 45 samples of bee-collected pollen were gathered from beekeepers who placed their bee colonies on monocultures of sunflower, rape and poppy fields within their flying distance. Bee pollen was collected from bees’ legs by special devices placed at the entrance to hives. Samples were examined for the concentration and identification of microscopic fungi able to grow on Malt and Czapek-Dox agar and mycotoxins content [deoxynivalenol (DON), T-2 toxin (T-2), zearalenone (ZON) and total aflatoxins (AFL), fumonisins (FUM), ochratoxins (OTA)] by direct competitive enzyme-linked immunosorbent assays (ELISA). The total number of microscopic fungi in this study ranged from 2.98 ± 0.02 in frozen sunflower bee pollen to 4.06 ± 0.10 log cfu.g^?1 in sunflower bee pollen after UV radiation. In this study, 449 isolates belonging to 21 fungal species representing 9 genera were found in 45 samples of bee pollen. The total isolates were detected in frozen poppy pollen 29, rape pollen 40, sunflower pollen 80, in dried poppy pollen 12, rape pollen 36, sunflower 78, in poppy pollen after UV radiation treatment 54, rape 59 and sunflower 58. The most frequent isolates of microscopic fungi found in bee pollen samples of all prevalent species were Mucor mucedo (49 isolates), Alternaria alternata (40 isolates), Mucor hiemalis (40 isolates), Aspergillus fumigatus (33 isolates) and Cladosporium cladosporioides (31 isolates). The most frequently found isolates were detected in sunflower bee pollen frozen (80 isolates) and the lowest number of isolates was observed in poppy bee pollen dried (12 isolates). The most prevalent mycotoxin of poppy bee pollen was ZON (361.55 ± 0.26 μg.kg^?1), in rape bee pollen T-2 toxin (265.40 ± 0.18 μg.kg^?1) and in sunflower bee pollen T-2 toxin (364.72 ± 0.13 μg.kg^?1) in all cases in frozen samples.