Cytogenetic evaluation and the association with polymorphisms of the CPY1A1 and NR1I3 genes in individuals exposed to BTEX

The gas station attendants are exposed daily to chemical agents that compose gasoline, such as BTEX (benzene, toluene, ethylbenzene, and xylene), and the exposure to these agents can cause a variety of effects on the human health. Among the various possible cell alterations associated with these exposures are the formation of micronuclei and of binucleated cells which are used as indicators of clastogenic action. Benzene, the main carcinogenic agent, is metabolized to more soluble forms and easily excreted by isoenzymes of cytochrome P450, such as CYP1A1. The CYP1A1 gene is highly polymorphic and one of its allele variations can be detected by the use of restriction endonucleasis MspI and is originated by the transition of a thymine by a cytosine (3798T>C), resulting in the polymorphic allele CYP1A1*2A. The objective of this study was to evaluate the cytogenetic damage induced by the exposure to BTEX and to associate it with the polymorphisms of the CYP1A1 and NR1I3 genes. Samples of exfoliated cells from the oral mucosa of 27 gas station attendants and from a control group were collected. The results found show that the group exposed to BTEX presents significantly higher alterations than those in the control group for micronuclei (MN; 6.85?±?1.33 vs. 2.96?±?1.91, P?<?0.001) and for the total of nuclear alterations observed (MN + binucleated cells (BNC); 9.59?±?4.73 vs. 5.07?±?2.21, P?<?0.001). When comparing the cytological alterations and the genotypes among the exposed individuals for the polymorphism 3798T>C of the CYP1A1 gene, homozygotes TT present MN + BNC significantly higher than carriers of the allele C (10.88?±?5.36 vs. 5.33?±?2.52, P?=?0.028). No association was observed in the control group or for the NR1I3 gene. These results show that molecular and cytogenetic data can be used in the future as tools to monitor individuals exposed to such compounds.     

Biochemical and standard toxic effects of acetaminophen on the macrophyte species Lemna minor and Lemna gibba

Acetaminophen is globally one of the most prescribed drugs due to its antipyretic and analgesic properties. However, it is highly toxic when the dosage surpasses the detoxification capability of an exposed organism, with involvement of an already described oxidative stress pathway. To address the issue of the ecotoxicity of acetaminophen, we performed acute exposures of two aquatic plant species, Lemna gibba and Lemna minor, to this compound. The selected biomarkers were number of fronds, biomass, chlorophyll content, lipid peroxidation (TBARS assay), and proline content. Our results showed marked differences between the two species. Acetaminophen caused a significant decrease in the number of fronds (EC₅₀?=?446.6 mg/L), and the establishment of a dose-dependent peroxidative damage in L. minor, but not in L. gibba. No effects were reported in both species for the indicative parameters chlorophyll content and total biomass. However, the proline content in L. gibba was substantially reduced. The overall conclusions point to the occurrence of an oxidative stress scenario more prominent for L. minor. However, the mechanisms that allowed L. gibba to cope with acetaminophen exposure were distinct from those reported for L. minor, with the likely involvement of proline as antioxidant.     

Occurrence of PCDD/Fs and dioxin-like PCBs in superficial sediment of Portuguese estuaries

Superficial sediments collected from seven estuarine systems located along the Portuguese coast were analyzed for 7 polychlorinated dibenzo-p-dioxins (PCDDs), 10 polychlorinated dibenzofurans (PCDFs), and 12 dioxin-like polychlorinated biphenyls (dl-PCBs). Total PCDD/F concentration ranged from 4.6 to 464 pg g^?1 dry weight (dw), while that of dl-PCBs varied from 26.6 to 8,693 pg g^?1 dw. In general, the highest PCDD/F and dl-PCB concentrations were associated with densely populated and industrially impacted areas. Additionally, PCDD/F revealed a predominance of octachlorodibenzodioxin (OCDD) to total PCDD/Fs, while PCB 118 was the major contributor to total dl-PCBs. This study provided a global perspective of the contamination status of Portuguese estuaries by dioxin-like compounds and allowed a comparison between the investigated systems and other systems worldwide. PCDD/F and dl-PCB levels found in the collected sediments were lower than those of highly impacted areas from different parts of the globe. Nevertheless, comparison with guidelines and quality standards from other countries indicated that some Portuguese estuarine areas with a high industrialization level present PCDD/F and dl-PCB concentrations in superficial sediment that may constitute a risk to aquatic organisms.     

Effects of chronic exposure to lead,copper, zinc,and cadmium on biomarkers of the European eel,Anguilla anguilla

Exposure to specific metallic compounds can cause severe deleterious modifications in organisms. Fishes are particularly prone to toxic effects from exposure to metallic compounds via their environment. Species that inhabit estuaries or freshwater environments can be chronically affected by persistent exposure to a large number of metallic compounds, particularly those released by industrial activities. In this study, we exposed yellow eels (European eel, Anguilla anguilla) for 28 days to environmentally relevant concentrations of four specific metals; lead (300, 600, and 1,200 μg/l), copper (40, 120, and 360 μg/l), zinc (30, 60, and 120 μg/l) and cadmium (50, 150, and 450 μg/l). The selected endpoints to assess the toxicological effects were neurotransmission (cholinesterasic activity in nervous tissue), antioxidant defense, and phase II metabolism (glutathione-S-transferase [GST] activity, in both gills and liver tissues), and peroxidative damage. The results showed an overall lack of effects on acetylcholinesterase for all tested metals. Lead, copper, and cadmium exposure caused a significant, dose-dependent, increase in GST activity in gill tissue. However, liver GST only significantly increased following zinc exposure. No statistically significant effects were observed for the thiobarbituric acid reactive substances assay, indicating the absence of peroxidative damage. These findings suggest that, despite the occurrence of an oxidative-based response after exposure to lead, copper, and cadmium, this had no consequence in terms of peroxidative membrane damage; furthermore, cholinergic neurotoxicity caused by lead, copper, and cadmium did not occur. The implications of these results are further discussed.     

Cholinesterase characterization in Corbicula fluminea and effects of relevant environmental contaminants: A pesticide (chlorfenvinphos) and a detergent (SDS)

This study characterizes the cholinesterasic forms present in the bivalve Corbicula fluminea, through the analysis of differential affinity towards various substrates and inhibitors. This approach also permitted the validation of cholinesterase inhibition in the mentioned species for monitoring purposes by evaluating the inhibitory activity of a detergent (SDS) and a pesticide (chlorfenvinphos), both in vitro and in vivo. The cholinesterasic form present in the tissues of C. fluminea showed intermediate properties in terms of its relation with substrates and inhibitors. Furthermore, it was not inhibited by the detergent SDS under any of the test conditions, but was clearly inhibited by the selected organophosphate. The data show clearly that the main cholinesterasic form in C. fluminea is responsive to the organophosphate chlorfenvinphos, thus supporting the use of this biomarker for the environmental monitoring of specific contaminants such as anticholinesterasic pesticides. Despite not being commonly attained in the wild, the levels of chlorfenvinphos required to elicit a significant ChE inhibition were only one order of magnitude above actual values of aquatic contamination by this pesticide.     

Use of the genus Artemia in ecotoxicity testing

Information related to varied uses of several species of the genus Artemia (commonly known as brine shrimp), is dispersed among literature from several scientific areas, such as Ecology, Physiology, Ecotoxicology, Aquaculture and Genetics. The present paper reviews information related to Artemia that may be considered relevant for ecotoxicity testing. Integration of different areas of scientific knowledge concerning biology, life cycle and environmental needs of Artemia is of crucial importance when considering the interpretation of results drawn from tests involving this genus. Furthermore, this paper provides suggestions to overcome problems related to toxicity assessment with the use of Artemia as test organism in bioassays, under the scope of estuarine, marine and hypersaline environments. Aspects related to variability in results, adoptable toxicity end-points, culture conditions, characteristics of species and strains, influence of geographical origins over physiological features and responses to exposure to chemical agents are considered.     

Association between biotic and abiotic parameters and the occurrence of cyanobacteria in a Brazilian reservoir

The water reservoir of Vargem das Flores, Brazil, has been subject to eutrophication problems. The occurrence of cyanobacterial blooms in this water body has raised concerns regarding the presence of toxic metabolites. In order to understand the factors that promote the proliferation and distribution of cyanobacteria in the reservoir, physicochemical parameters, species composition, and toxicity of the reservoir water column were analyzed at monthly intervals over a 3-year period (2004-2007) using a range of analytical techniques. The results showed that cyanobacteria were absent in the dry period, returning after the onset of stratification. A succession pattern was observed within this group. The Aphanocapsa genus was dominant in the last rainy period of the study (October 2006 to March 2007). However toxicity was only detected when the genera Microcystis and Sphaerocavum were present (October 2005). The principal components analysis showed negative correlation between cyanobacteria and orthophosphate. The change of dominant cyanobacteria along the 3 years of study and the final prevalence of nontoxic genera in the reservoir is an evidence of water quality improvement in response to the management techniques implemented in the reservoir basin.     

Effects of anticholinesterase drugs on biomarkers and behavior of pumpkinseed, Lepomis gibbosus (Linnaeus, 1758)

The presence of pharmaceutical residues in the aquatic environment has recently received great attention, as potential adverse effects may arise from their presence. Inhibition of cholinesterases (ChE) has been widely used as an environmental biomarker of exposure to organophosphates (OP) and carbamate (CB) pesticides. However, other widespread anthropogenic contaminants - including pharmaceutical drugs - can exert toxic effects through ChE inhibition. Studies with aquatic species have shown that inhibition of ChE is associated with behavioral changes. Bearing this in mind, this work aimed to study the effects on individual behavior and acetylcholinesterase (AChE) activity of selected tissues of Lepomis gibbosus, after exposure to the anticholinesterasic drugs neostigmine and pyridostigmine. Results revealed that neostigmine significantly decreased the activity of AChE in the head (LOEC = 100 mg L(-1)), but not in dorsal muscle. On the other hand, pyridostigmine significantly decreased the activity of AChE in both the head (LOEC = 0.001 mg L(-1)) and dorsal muscle homogenates (LOEC = 100 mg L(-1)). The impairment of this enzymatic form was attained at ecologically relevant concentrations (in the case of pyridostigmine, for head AChE), which is an interesting finding, specially considering that these toxic effects occurred for a pharmaceutical compound. Contrarily, there were no significant differences in the behavior of L. gibbosus in any parameter, for neither drug. These results suggest that the behavioral parameters analyzed (scototaxis and lethargy) in L. gibbosus could not be regarded as suitable markers to assess the effects of drugs such as neostigmine and pyridostigmine. In contrast, the pattern of response elicited by cholinesterasic inhibition showed the usefulness of this toxicological parameter for the assessment of pharmaceuticals in the environment, namely anticholinesterasics.     

Influence of the composition of the initial mixtures on the chemical composition,physicochemical properties and humic-like substances content of composts

The influence of the proportion of C- and N-rich raw materials (initial C/N ratio) and bulking agent on the chemical functional groups composition, humic-like substances (HS-like) content and physicochemical properties of composts was assessed. To achieve these goals, seven initial mixtures (BA1–6 and C1) of dog food (N-rich raw material) were composted with wheat flour (C-rich raw material). Composts were analyzed in terms of chemical functional groups, physicochemical, maturity and stability parameters.The C-rich raw material favored the formation of oxidized organic matter (OM) during the composting process, as suggested by the variation of the ratios of the peaks intensity of FT-IR spectra, corresponding to a decrease of the polysaccharides and an increase of aromatic and carboxyl-containing compounds. However, although with high proportion of C-rich raw material, mixtures with low initial C/N seems to have favored the accumulation of partially oxidized OM, which may have contributed to high electrical conductivity values in the final composts. Therefore, although favoring the partial transformation of OM into stabilized HS-like, initial mixtures with high proportion of C-rich raw material but with low initial C/N led to unstable composts.On the other hand, as long as a high percentage of bulking agent was used to promote the structure of biomass and consequently improve of the aeration conditions, low initial C/N was not a limiting factor of OM oxidation into extractable stabilized humic-like acids.