Assessment of oxidative stress due to exposure to pesticides in plasma and urine of traditional limited-resource farm workers: Formation of the DNA-adduct 8-hydroxy-2-deoxy-guanosine (8-OHdG)

The objective of this study was to assess the risk of genotoxicity caused due to oxidative stress using plasma and urinary levels of 8-hydroxy-2-deoxyguanosine (8-OHdG) in farm workers for six months during a growing season. Blood and urine samples were collected once a month for six months (June to November 2003) from farm workers (n = 15) and urban unexposed controls (n = 10). Plasma and urinary levels of 8-OHdG were evaluated by Enzyme Linked Immunosorbent Assay (ELISA) technique. There was no significant difference in the urinary levels of 8-OHdG between the farm workers and the control group, but there was an approximately four-fold increase in mean values of plasma 8-OHdG levels in the farm worker group (P ≤ 0.05).     

In vitro methylation of DNA by the fumigant methyl bromide

Abstract

Investigations have shown that the physical state of the DNA strongly influences the pattern of methylation observed when DNA or a substance containing DNA is treated with the fumigant methyl bromide. 1‐Methyl‐adenine and 7‐methylguanine were identified, after hydrolysis, as the major methylated bases of DNA which had been treated in the solid state. 3‐Methylcytosine, 3‐methyladenine and 7‐methyladenine were found as minor products. The overall methylation pattern was similar to that observed earlier for DNA of maize and wheat which had been fumigated. In contrast, when buffer solutions of DNA were treated with methyl bromide, the N‐7 position of guanine was the major site and the N‐3 position of adenine was the second most important site of methylation. This result corresponds to that previously observed in similar studies with buffer solutions of DNA and other methylating agents.     

Determination of hypoxia and dietary copper mediated sub-lethal toxicity in carp, Cyprinus carpio, at different levels of biological organisation

Hypoxic events frequently occur in the aquatic environment in association with micro pollutants, including heavy metals. Only a few studies are however available on the uptake and biological responses of heavy metals under hypoxic conditions. To elucidate the phenomenon, mirror carp Cyprinus carpio L. (16.13-16.22 g) were exposed chronically to dietary copper (Cu; 250 and 500 mg kg dry wt.⁻¹) for 30 d under normoxic (8.25 mg O₂ L⁻¹) and hypoxic (∼3 mg O₂ L⁻¹) conditions and adopting an integrated approach, sub-lethal biomarker responses were determined at different levels of biological organisation. Level of oxidative DNA damage (as determined by modified Comet assay) showed strong significant difference following exposure to dietary Cu level under normoxic (1.6-fold) as well as under hypoxic condition at both Cu levels (2.1 and 2.5-folds respectively). Significant difference was also observed for haematological parameters (i.e. increased red and white blood cells, haematocrit value and haemoglobin concentration). Quantitative histology revealed alterations in tissues (i.e. liver and gills) for hypoxic and all dietary Cu treatment groups under both normoxic and hypoxic conditions suggesting a compensatory response to these organs (p < 0.05). The order of Cu accumulation in tissues (as determined by ICP-OES) was liver > intestine > kidney > gill. Interestingly, SGR under both normoxic and hypoxic conditions reduced with elevating Cu levels (p = 0.019). Overall, the results provide evidence for enhanced toxicological responses in fish following exposure to Cu either alone or in combination with hypoxic condition and lends support to the evolving viewpoint that many water quality guidelines should be revisited in terms of new ecotoxicological criteria.     

Cadmium-induced DNA damage and mutations in Arabidopsis plantlet shoots identified by DNA fingerprinting

Random amplified polymorphic DNA (RAPD) test is a feasible method to evaluate the toxicity of environmental pollutants on vegetal organisms. Herein, Arabidopsis thaliana (Arabidopsis) plantlets following Cadmium (Cd) treatment for 26 d were screened for DNA genetic alterations by DNA fingerprinting. Four primers amplified 20-23 mutated RAPD fragments in 0.125-3.0 mg L⁻¹ Cd-treated Arabidopsis plantlets, respectively. Cloning and sequencing analysis of eight randomly selected mutated fragments revealed 99-100% homology with the genes of VARICOSE-Related, SLEEPY1 F-box, 40S ribosomal protein S3, phosphoglucomutase, and noncoding regions in Arabidopsis genome correspondingly. The results show the ability of RAPD analysis to detect significant genetic alterations in Cd-exposed seedlings. Although the exact functional importance of the other mutated bands is unknown, the presence of mutated loci in Cd-treated seedlings, prior to the onset of significant physiological effects, suggests that these altered loci are the early events in Cd-treated Arabidopsis seedlings and would greatly improve environmental risk assessment.     

The adsorption of short single-stranded DNA oligomers to mineral surfaces

We studied the adsorption of short single-stranded deoxyribonucleic acid (ssDNA) oligomers, of approximately 30 nucleotides (nt) in length, of varying sequence, adenine + guanine + cytosine (AGC) content, and propensity to form secondary structure, to equal surface area samples of olivine, pyrite, calcite, hematite, and rutile in 0.1 M NaCl, 0.05 M pH 8.1 KHCO₃ buffer. Although the mineral surfaces have widely varying points of zero charge, under these conditions they show remarkably similar adsorption of ssDNA regardless of oligomer characteristics. Mineral surfaces appear to accommodate ssDNA comparably, or ssDNA oligomers of this length are able to find binding sites of comparable strength and density due to their flexibility, despite the disparate surface properties of the different minerals. This may partially be due charge shielding by the ionic strength of the solutions tested, which are typical of many natural environments. These results may have some bearing on the adsorption and accumulation of biologically derived nucleic acids in sediments as well as the abiotic synthesis of nucleic acids before the origin of life.     

New Genetic Approach to Detecting Individuals of Rare and Endangered Species

Abstract: Many rare and endangered species are difficult to locate, observe, and study. Consequently, many individuals, breeding pairs, and even populations of such species could remain undetected. Genetic markers can potentially be used to detect the existence of undiscovered individuals and populations, and we propose a method to do so that requires 3 conditions. First, sampling of the known population(s) of the target species must be comprehensive. Second, the species must display a reasonable level of philopatry and genetic structuring. Third, individuals must be able to be caught outside of breeding locations (e.g., at courtship or feeding areas, in flight), and the level of recapture must be reasonably high. We applied our method to the Chatham Island Taiko (Pterodroma magentae), one of the world's most endangered seabirds. We sequenced the Taiko mitochondrial cytochrome b gene and both copies of a fragment of the duplicated domain I of the control region. Twenty‐one haplotypes were revealed, including 4 (19%) not found in birds at known burrows. These results suggest there are more burrow groups yet to be located. The species is a pelagic gadfly petrel that inhabits land only in the breeding season during which it is nocturnal and nests in burrows. Taiko burrows are situated in dense forest in a remote area of Chatham Island, and are consequently difficult to locate and study. It is important that all Taiko burrows be discovered to enable monitoring and protection of the birds from exotic predators.