Planarian neoblast micronucleus assay for evaluating genotoxicity

Planarian neoblasts are somatic stem cells that have the potential to be used in genotoxicity assays due to their proliferative nature, sensitivity to genotoxic agents, and experimental accessibility. Two freshwater planarian species, Girardia tigrina and Girardia schubarti, were used to develop a neoblast-based micronucleus (MN) assay to assess genotoxicity. Intact or regenerating planarians were exposed to gamma-rays, methyl methanesulphonate (MMS), or cyclophosphamide (CP), and neoblast MN frequency was measured. Exposure to the clastogens had no detectable effect on the MN frequency of intact planarian neoblasts. However, for regenerating individuals, active neoblast proliferation was induced by decapitation, and gamma-ray doses as low as 0.5 Gy, and MMS and CP concentrations as low as 0.8 microM and 100 mM, respectively, induced a significant increase in MN frequency. Exposure to higher doses of gamma-rays consistently resulted in detectable increases in MN frequency. For MMS and CP, concentrations of up to 1.6 microM and 200 mM, respectively, resulted in significant increases in MN frequency, but exposures to higher concentrations led to a decrease to non-significant levels, possibly due to cytotoxic effects of MMS and CP. After completion of regeneration, the MN frequencies returned to those of non-exposed controls, indicating that the neoblast MN assay for regenerating G. tigrina or G. schubarti reflects chromosomal damage caused by acute exposure to clastogenic agents. Upon standardization, this assay may represent an interesting alternative that allows damage caused to freshwater organisms by potentially genotoxic environmental pollutants to be monitored.     

Mercury reduction studies to facilitate the thermal decontamination of phosphor powder residues from spent fluorescent lamps

This work investigates the thermal release of mercury from phosphor powder of spent fluorescent lamps. The treatment conditions and the ability of various reducing agents (primarily sodium borohydride) to lower the overall heating temperature required to improve the release of Hg have been evaluated. Hg species in samples were monitored in a thermal desorption atomic absorption spectrometer system, and total mercury was analyzed in a cold vapor atomic absorption spectrometer. Sodium borohydride was the best reducing agent among the ones studied. However, citric acid presented a high capacity to weaken mercury bonds with the matrix. When the sample was crushed with sodium borohydride for 40min in a mass ratio of 10:1 (sample:reducing agent) and submitted to thermal treatment at 300 degrees C for 2h, the concentration of mercury in a phosphor powder sample with 103mgkg(-1) of mercury reached 6.6mgkg(-1).     

Influence of population,income and electricity consumption on per capita municipal solid waste generation in São Paulo State,Brazil

Journal of Material Cycles and Waste Management - Predicting municipal solid waste (MSW) generation is fundamental in choosing and scaling the processes involved in municipal management. The...     

Occupational exposure of farm workers to pesticides: biochemical parameters and evaluation of genotoxicity

To assess the effects of exposure to complex mixtures of pesticides in farm workers from two communities from Rio Grande do Sul, Brazil, we evaluated the activities of butyrylcholinesterase (BChE) and delta-aminolevulinic acid dehydratase (ALA-D) enzymes, hematological, lipid parameters, and genotoxicity using two tests to detect DNA damage, the Comet assay in peripheral blood leukocytes and the micronucleus (MN) test in oral mucosa cells. The use of personal protective equipment (PPE), age and smoke habits were considered in the analysis. There was a significant decrease in the BChE and ALA-D activities in farm workers (n=37) relative to the control group (n=20) (P< or =0.05 and P< or =0.001, respectively). The Comet assay in peripheral blood leukocytes showed that the Damage index and Damage frequency observed in the exposed group were significantly higher in relation to the controls (P< or =0.001, and P< or =0.05, respectively). No differences were detected regarding the hematological parameters, lipids profile, and MN frequencies. In addition, no significant differences were observed between younger (< or =38 years) and older subjects (>38 years), or between smokers and non-smokers within the groups, either by Comet assay or MN test. However, the use of PPE seems to be important in the prevention of contamination, as suggested by BChE levels and Comet assay results.     

Pine needles as passive bio-samplers to determine polybrominated diphenyl ethers

Eight polybrominated diphenyl ethers (PBDEs) were determined in pine needles of three species (Pinus halepensis, Pinus pinea and Pinus nigra) collected in the NE Spain in an attempt to use this matrix for the biomonitoring of airborne PBDEs. The method used was based in ultrasonic extraction followed by alumina and Florisil clean-up and determination by gas chromatography coupled to mass spectrometry in negative chemical ionization. Recoveries were between 99% and 138%, limits of detection between 0.011 and 0.070 ng g⁻¹-dw (0.232 ng g⁻¹-dw for BDE 209) and repeatability lower than 13%. PBDE levels ranged between 0.027 ng g⁻¹-dw and 13.04 ng g⁻¹-dw, with predominance of BDE 209, followed by BDEs 47. P. halepensis was the species with the highest PBDE levels and P. nigra, the least, according to their widespread and remote distribution, respectively. The presence of PBDEs in pine needles was attributed to the release of in-use PBDEs, transport through atmosphere and adsorption upon lipid rich pine needles. Given the easy collection of pine needles, its ample distribution and its potential to accumulate airborne contaminants, this matrix is proposed as passive bio-sampler to be used in PBDE monitoring programs.

Capsule

Pine needles can be used to biomonitor airborne PBDEs.     

Optimization of animal manure vermicomposting based on biomass production of earthworms and higher plants

The goal of this study was to optimize the mixture of swine manure (SM) and cattle manure (CM) used in the vermicomposting process, seeking to increase the manure biodegradation rate and enhance the biomass production of both earthworms and higher plants. To achieve this goal, physico-chemical parameters were determined to assess the final compost quality after 50 days of vermicomposting. The different manure ratios used to produce the composts (C) were as follows (SM:CM, % m/m basis): C1 100:0, C2 (75:25), C3 (50:50), C4 (25:75), and C5 (0:100). In addition, the earthworm biomass and the phytoproductivity of lettuce (Lactuca sativa L.) plants grown in mixtures (1:1) of natural soil and the most viable vermicomposts were investigated. The C1 and C2 compost compositions were associated with high earthworm mortality rates. The C3 compost provided the highest mineral concentrations and C5 showed the highest lettuce yield (wet biomass). The results verify that stabilized cattle manure is an excellent substrate for the vermicomposting process and that fresh swine manure must be mixed with pre-stabilized cattle manure to ensure an optimized vermicomposting process, which must be controlled in terms of temperature and ammonia levels. It is concluded that small livestock farmers could add value to swine manure by applying the vermicomposting process, without the need for high investments and with a minimal requirement for management of the biodegradation process. These are important technical aspects to be considered when circular economy principles are applied to small farms.     

Phylogenetic diversity and functional characterization of the Manila clam microbiota: a culture-based approach

According to the hologenome theory, the microbiota contributes to the fitness of the holobiont having an important role in its adaptation, survival, development, health, and evolution. Environmental stress also affects the microbiota and its capability to assist the holobiont in coping with stress factors. Here, we analyzed the diversity of cultivable bacteria associated with Manila clam tissues (mantle, gills, hemolymph) in two non-contaminated sites (Portugal and France) and one metal-contaminated site (Portugal). A total of 240 isolates were obtained. Representative isolates (n = 198) of the overall diversity were identified by 16S rDNA sequencing and subjected to functional characterization. Isolates affiliated with Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes. Proteobacteria (mostly Pseudoalteromonadaceae and Vibrionaceae) were dominant in non-contaminated sites while Actinobacteria (mostly Microbacteriaceae) dominated in the metal-contaminated site. The main factor affecting the microbiota composition was contamination. No significant differences were observed between clam tissues and geographic regions. Several isolates tested positive for antibacterial activity, biofilm formation, protease, and siderophore production. The results show that the Manila clam harbors a diverse microbiota that may contribute to clam protection and overall fitness, as well as to its adaptation to stressful environments. In addition, the Manila clam microbiota is revealed as a promising source of novel probiotics with potential application in aquaculture.     

Method of median semi-variance for the analysis of left-censored data: Comparison with other techniques using environmental data

In environmental monitoring, variables with analytically non-detected values are commonly encountered. For the statistical evaluation of these data, most of the methods that produce a less biased performance require specific computer programs. In this paper, a statistical method based on the median semi-variance (SemiV) is proposed to estimate the position and spread statistics in a dataset with single left-censoring. The performances of the SemiV method and 12 other statistical methods are evaluated using real and complete datasets. The performances of all the methods are influenced by the percentage of censored data. In general, the simple substitution and deletion methods showed biased performance, with exceptions for L/2, Inter and L/√2 methods that can be used with caution under specific conditions. In general, the SemiV method and other parametric methods showed similar performances and were less biased than other methods. The SemiV method is a simple and accurate procedure that can be used in the analysis of datasets with less than 50% of left-censored data.     

Analytical methods and validation for determining trace elements in red wines

The objective of this work is to quantify As, Hg, Cd, Ni and Pb in Portuguese red wines. First, the methods for the quantification of trace elements in red wines were validated. Several pre-treatments were compared, namely a pre-digestion process with HNO₃, a pre-oxidation step with H₂O₂, and a spiking step of wine samples with a known concentration of the trace elements analyzed. Except for As, it was determined that the quantification of the trace elements does not require a pre-digestion process with HNO₃. For all of the trace elements analyzed, a pre-oxidation step with H₂O₂ may enable an accurate quantification. The techniques chosen for the quantification of trace elements were hydride generation atomic absorption spectrometry (HGAAS) for As and Hg, electrothermal atomic absorption spectrometry (ETAAS) for Cd, and flame atomic absorption spectrometry (FAAS) for Ni and Pb. In the second stage of this work, 25 Portuguese red wines spanning all of the red wine-producing regions were analyzed for all of the five trace elements referred to above. Only Cd and Pb have shown concentrations above the limit values defined by the “Organization Internationale de la Vigne et du Vin.” The Target Hazard Quotient (THQ) equation was used to determine in which wine-producing regions that wine consumption can be a problem for public health in terms of the concentrations of the five trace elements analyzed. THQ values have indicated that for the universe of the 25 red wines analyzed no region produces wines that can pose problems for public health, when the Portuguese red wine standard consumption is considered.     

Assessment of metolachlor and diuron leaching in a tropical soil using undisturbed soil columns under laboratory conditions

In the present study, diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea] and metolachlor [2-chloro-N-(2-ethyl-6-methylphenyl)-N-(2-metoxi-1-methylethyl)acetamide] leaching was studied in undisturbed soil columns collected in a cotton crop area in Mato Grosso State, Brazil. The pesticides were applied to the soil surface in dosages similar to those used in a cotton plantation. To assess the leaching process, soil columns were submitted to simulated rain under laboratory conditions at 25 ± 3°C, in the absence of wind and direct solar radiation. During the rain simulations, leachate solutions were collected and herbicide concentrations were determined. At the end of the experiment, the soil columns were cut into 10 cm sections to determine the remaining herbicide concentrations through the soil profile. Metolachlor was detected in all soil sections, and approximately 4% of the applied mass was leached. Diuron was detected only in the upper two soil sections and was not detected in the leachate. A linear correlation (r > 0.94) between the metolachlor soil concentrations and the organic contents of the soil sections was observed. Mass balance suggests that around 56% of diuron and 40% of metolachlor were degraded during the experiments. Measurements of the water table depth in the area where the samples were collected showed that it varied from 2 to 6 m and is therefore vulnerable to contamination by the studied herbicides, particularly metolachlor, which demonstrated a higher leaching potential.