Frequency of micronuclei and of other nuclear abnormalities in erythrocytes of the grey mullet from the Mondego,Douro and Ave estuaries—Portugal

Fish are bioindicators of water pollution, and an increased rate of their erythrocyte nuclear morphological abnormalities (ENMAs)—and particularly of erythrocyte micronuclei (EMN)—is used as a genotoxicity biomarker. Despite the potential value of ENMAs and MN, there is scarce information about fish captured in Iberian estuaries. This is the case of the Portuguese estuaries of the Mondego, Douro and Ave, suffering from different levels of environmental stress and where chemical surveys have been disclosing significant amounts of certain pollutants. So, the aim of this study was to evaluate genotoxicants impacts and infer about the exposure at those ecosystems, using the grey mullet (Mugil cephalus) as bioindicator and considering the type and frequency of nuclear abnormalities of erythrocytes as proxies of genotoxicity. Sampling of mullets was done throughout the year in the important Mondego, Douro and Ave River estuaries (centre and north-western Portugal). The fish (total n?=?242) were caught in campaigns made in spring–summer and autumn–winter, using nets or fishing rods. The sampled mullets were comparable between locations in terms of the basic biometric parameters. Blood smears were stained with Diff-Quik to assess the frequencies of six types of ENMAs and MN (given per 1,000 erythrocytes). Some basic water physicochemical parameters were recorded to search for fluctuations matching the ENMAs. Overall, the most frequent nucleus abnormality was the polymorphic type, sequentially followed by the blebbed/lobed/notched, segmented, kidney shaped, vacuolated, MN and binucleated. The total average frequency of the ENMAs ranged from 73?‰ in the Mondego to 108?‰ in the Ave. The polymorphic type was typically ≥50 % of the total ENMAs, averaging about 51?‰, when considering all three estuaries. The most serious lesion—the MN—in fish from Mondego and Douro had a similar frequency (≈0.38?‰), which was significantly lower than that in the Ave (0.75?‰). No significant seasonal differences existed as to the MN rates and seasonal differences existed almost only in the Douro, with the higher values in AW. In general, the pattern of ENMAs frequencies was unrelated with the water physicochemical parameters. Considering the data for both the total ENMAs and for each specific abnormality, and bearing in mind that values of MN in fish erythrocytes >0.3?‰ usually reflect pollution by genotoxicants, it is suggested that mullets were likely being chronically exposed to such compounds, even in the allegedly less polluted ecosystem (Mondego). Moreover, data supported the following pollution exposure gradient: Mondego?<?Douro?<?Ave. The scenario and inferences nicely agree with the published data from chemical monitoring.     

Correlations between PAH bioavailability,degrading bacteria,and soil characteristics during PAH biodegradation in five diffusely contaminated dissimilar soils

The natural biodegradation of seven polycyclic aromatic hydrocarbons (PAHs) by native microorganisms was studied in five soils from Normandy (France) from diffusely polluted areas, which can also pose a problem in terms of surfaces and amounts of contaminated soils. Bioavailability tests using cyclodextrin-based extractions were performed. The natural degradation of low molecular weight (LMW) PAHs was not strongly correlated to their bioavailability due to their sorption to geosorbents. Conversely, the very low degradation of high molecular weight (HMW) PAHs was partly correlated to their poor availability, due to their sorption on complexes of organic matter and kaolinites or smectites. A principal component analysis allowed us to distinguish between the respective degradation behaviors of LMW and HMW PAHs. LMW PAHs were degraded in less than 2–3 months and were strongly influenced by the relative percentage of phenanthrene-degrading bacteria over total bacteria in soils. HMW PAHs were not significantly degraded, not only because they were less bioavailable but also because of a lack of degrading microorganisms. Benzo[a]pyrene stood apart since it was partly degraded in acidic soils, probably because of a catabolic cooperation between bacteria and fungi.     

Strategies to evaluate biodegradability: application to chlorinated herbicides

The biodegradability of nitrochlorinated (diuron and atrazine) and chlorophenoxy herbicides (2,4-D and MCPA) has been studied through several bioassays using different testing times and biomass/substrate ratios. A fast biodegradability test using unacclimated activated sludge yielded no biodegradation of the herbicides in 24 h. The inherent biodegradability test gave degradation percentages of around 20–30 % for the nitrochlorinated herbicides and almost complete removal of the chlorophenoxy compounds. Long-term biodegradability assays were performed using sequencing batch reactor (SBR) and sequencing batch membrane bioreactor (SB-MBR). Fixed concentrations of each herbicide below the corresponding EC₅₀ value for activated sludge were used (30 mg L^?1 for diuron and atrazine and 50 mg L^?1 for 2,4-D and MCPA). No signs of herbicide degradation appeared before 35 days in the case of diuron and atrazine and 21 days for 2,4-D, whereas MCPA was partially degraded since the early stages. Around 25–36 % degradation of the nitrochlorinated herbicides and 53–77 % of the chlorophenoxy ones was achieved after 180 and 135 days, respectively, in SBR, whereas complete disappearance of 2,4-D was reached after 80 days in SB-MBR.     

Using multivariate regression modeling for sampling and predicting chemical characteristics of mixed waste in old landfills

Municipal solid waste landfills pose a threat on environment and human health, especially old landfills which lack facilities for collection and treatment of landfill gas and leachate. Consequently, missing information about emission flows prevent site-specific environmental risk assessments. To overcome this gap, the combination of waste sampling and analysis with statistical modeling is one option for estimating present and future emission potentials. Optimizing the tradeoff between investigation costs and reliable results requires knowledge about both: the number of samples to be taken and variables to be analyzed.This article aims to identify the optimized number of waste samples and variables in order to predict a larger set of variables. Therefore, we introduce a multivariate linear regression model and tested the applicability by usage of two case studies. Landfill A was used to set up and calibrate the model based on 50 waste samples and twelve variables. The calibrated model was applied to Landfill B including 36 waste samples and twelve variables with four predictor variables.The case study results are twofold: first, the reliable and accurate prediction of the twelve variables can be achieved with the knowledge of four predictor variables (Loi, EC, pH and Cl). For the second Landfill B, only ten full measurements would be needed for a reliable prediction of most response variables. The four predictor variables would exhibit comparably low analytical costs in comparison to the full set of measurements. This cost reduction could be used to increase the number of samples yielding an improved understanding of the spatial waste heterogeneity in landfills.Concluding, the future application of the developed model potentially improves the reliability of predicted emission potentials. The model could become a standard screening tool for old landfills if its applicability and reliability would be tested in additional case studies.     

Natural Products: Repellency and Toxicity of Wild Tomato Leaf Extracts to the Two-Spotted Spider Mite,Tetranychus urticae Koch

The potential of using phytochemicals from leaves of wild tomato for controlling the two-spotted spider mite, Tetranychus urticae Koch, is explored in this study as a promising alternative to the use of synthetic pesticides. Wild tomato accessions of Lycopersicon hirsutum plants that are not consumed by humans were planted under greenhouse conditions for mass production of leaves. Crude extracts from leaves of three accessions of L. hirsutum, six accessions of L. hirsutum f. glabratum, and one accession each of L. pennellii and L. pimpinellifolium were prepared in chloroform, ethanol and hexane. Two spider mite bioassays, one a measure of antibiosis and the other a measure of repellency, were utilized to determine the acaricidal performance of the crude extracts. The bioassay for antibiosis was a 6-h no-choice test. The bioassay for repellency utilized a ring bioassay. Chloroform leaf extracts of L. hirsutum f. glabratum accessions (PI-251304, PI-134417, PI-134418, and PI-126449) exhibited greatest antibiotic activity on two-spotted spider mites; the hexane extracts exhibited greatest repellency. Extracts from PI-251304, PI-126449, PI-134417, and PI-134418 were especially lethal (chloroform) or repellent (hexane). We investigated differences in chemical composition of the crude leaf extracts that may explain the observed differences in mortality and repellency among the different accessions. Major chemical compounds (α -curcumene, α -zingiberene, trans-caryophyllene, 2-undecanone, and 2-tridecanone) known to have pesticidal efficacy were detected and quantified in the crude leaf extracts using a gas chromatograph (GC) equipped with a mass spectrometer (GC/MS). Lethality of extracts was mainly associated with the presence of high concentrations of 2-tridecanone; repellency of extracts was mainly associated with the presence of trans-caryophyllene. Leaf extracts of L. hirsutum f. glabratum accessions that contain significant quantities of 2-tridecanone and/or trans-caryophyllene could be useful for managing populations of spider mites, which could reduce reliance on synthetic pesticides.     

Screening Capsicum Accessions for Capsaicinoids Content

Ninety Capsicum accessions selected from the USDA Capsicum germplasm collection were screened for their capsaicinoids content using gas hromatography with nitrogen phosphorus detection (GC/NPD). Fresh fruits of Capsicum chinense, C. frutescens, C. baccatum, C. annuum, and C. pubescens were extracted with methanol and analyzed for capsaicin, dihydrocapsaicin, and nordihydrocapsaicin. Mass spectrometry of the fruit crude extracts indicated that the molecular ions at m/z 305, 307, and 293, which correspond to capsaicin, dihydrocapsaicin, and nordihydrocapsaicin, respectively, have a common benzyl cation fragment at m/z 137 that can be used for monitoring capsaicinoids in pepper fruit extracts. Capsaicin and dihydrocapsaicin were the dominant capsaicinoids detected. Capsaicin concentrations were typically greater than dihydrocapsaicin. Concentrations of total capsaicinoids varied from not detectable to 11.2 mg fruit^?1. Statistical analysis revealed that accession PI-441624 (C. chinense) had the highest capsaicin content (2.9 mg g^?1 fresh fruit) and accession PI-497984 (C. frutescens) had the highest dihydrocapsaicin content (2.3 mg g^?1 fresh fruit). Genebank accessions PI-439522 (C. frutescens) and PI-497984 contained the highest concentrations of total capsaicinoids.     

Antioxidants in Capsicum chinense: Variation among countries of origin

The main objective of this investigation was to evaluate fruits of C. chinense accessions for their concentration of β -carotene, ascorbic acid, and phenols for use as parents in breeding for these phytochemicals. Mature fruits of 63 accessions of C. chinense originally acquired from Belize, Brazil, Colombia, Ecuador, Mexico, Peru, Puerto Rico, and the United States were analyzed for their chemical composition. Fruits of C. chinense accessions PI-152452 (Brazil) and PI-360726 (Ecuador) contained the greatest concentrations of ascorbic acid (1.2 and 1.1 mg g^?1 fresh fruit, respectively), while PI-438648 (Mexico) contained the greatest concentration of total phenols content (349 μ g g^?1 fresh fruit) among the other 63 accessions tested. Accession PI-355817 from Ecuador contained the greatest concentrations of β -carotene (8 mg g^?1fresh fruit). These accessions were identified as potential candidates for mass production of antioxidants with health-promoting properties.     

Evaluation of protective effects of sulforaphane on DNA damage caused by exposure to low levels of pesticide mixture using comet assay

The objective of the study was to evaluate the potential risk of DNA damage due to exposure to a mixture of the most widely used pesticides, namely endosulfan, chlorpyriphos and thiram at an environmentally relevant concentration (5 μM each) and the DNA protective capacity of sulforaphane (SFN) (10–30 μg/mL). DNA damage in human lymphocytes was ascertained with Single Cell Gel Electrophoresis (SCGE), also called Comet Assay. For positive control, H₂O₂ at 100 mM was used. The pesticide mixture produced DNA damage at the concentration used in the lymphocytes. SFN was able to offer a statistically significant (P < 0.01), concentration-dependant protection to DNA damage between 10–20 μg/mL in both the pre-incubation and co-incubation strategies. The results indicate that exposure to low levels of these pesticide mixtures can induce DNA damage, and the presence of SFN in diet may reduce the incidence of genetic damage, especially in farm workers. However, it is not clear whether SFN is involved in quenching of the free radicals generated by the pesticide mixture or it is involved in DNA repair mechanism.     

Use of cholinesterase activity in monitoring chlorpyrifos exposure of steer cattle after topical administration

The aim of this work was to study the pharmacokinetic behavior and the inhibitory effect of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activities of chlorpyrifos (CPF) in steer cattle after pour-on administration. Determination of cholinesterase activity in plasma and erythrocyte was carried out according to Ellman kinetic method. CPF was analyzed by gas chromatography. AChE was the predominant form of cholinesterase analyzed, with low levels of BChE in plasma. Following the treatment with CPF, the maximum inhibitory effect on AChE or BChE were 50.88 ± 11.57 and 42.66 ± 12.01%, respectively. The chlorpyrifos plasma concentrations observed were low and they presented a high variability. Chlorpyrifos peak plasma concentration (10.42 ± 4.76 μ g/L) was reached at 8.42 ± 13.97 h. The pesticide was not detected in plasma after 48 h post treatment. The values of area under the curve (AUC) were 118.48 ± 87.46 μ g· h/L and mean resistance time (MRT) were 13.38 ± 10.41 h. The pour-on exposure to the organophosphate chlorpyrifos significantly reduced AChE and BChE activity in steer cattle and the recovery was not reached on 50 days post-treatment.     

Treatment of wastewater from red and tropical fruit wine production by zeolite anaerobic fluidized bed reactor

A study of the anaerobic treatment of wastewaters derived from red (RWWW) and tropical fruit wine (TFWWW) production was carried out in four laboratory-scale fluidized bed reactors with natural zeolite as bacterial support. These reactors operated at mesophilic temperature (35°C). Reactors R1 and R2 contained Chilean natural zeolite, while reactors R3 and R4 used Cuban natural zeolite as microorganism support. In addition, reactors R1 and R3 processed RWWW, while reactors R2 and R4 used TFWWW as substrate. The biomass concentration attached to zeolites in the four reactors studied was found to be in the range of 44–46 g volatile solids (VS)/L after 90 days of operation time. Both types of zeolites can be used indistinctly in the fluidized bed reactors achieving more than 80%–86% chemical oxygen demand (COD) removals for organic loading rates (OLR) of up to at least 20 g COD/L d. pH values remained within the optimal range for anaerobic microorganisms for OLR values of up to 20 and 22 g COD/L d for RWWW and TFWWW, respectively. Toxicity and inhibition levels were observed at an OLR of 20 g COD/L d in reactors R1 and R3 while processing RWWW, whereas the aforementioned inhibitory phenomena were not observed at an OLR of 24 g COD/L d in R2 and R4, treating TFWWW as a consequence of the lower phenolic compound content present in this substrate. The volatile fatty acid (VFA) levels were always lower in reactors processing TFWWW (R2 and R4) and these values (< 400 mg/L, as acetic acid) were lower than the suggested limits for digester failure. The specific methanogenic activity (SMA) was twice as high in reactors R2 and R4 than in R1 and R3 after 120 days of operation when all reactors operated at an OLR of 20 g COD/L d.