Exploring the binding mechanism and adverse toxic effects of persistent organic pollutant (dicofol) to human serum albumin: A biophysical, biochemical and computational approach.

The organochlorine pesticide dicofol (DCF), a persistent organic pollutant, is used as acaricide worldwide. Considering its large consumption in the agriculture sector and potential toxic effects such as endocrine disruption, carcinogenicity, and environmental persistence are detrimental to human health. To take an extensive evaluation of its potential toxicity, the current study was aimed to explore the binding mechanism and adverse effect of DCF on human serum albumin (HSA) by using an array of biophysical techniques (UV-visible, fluorescence, 3D fluorescence, and circular dichroism spectroscopy), isothermal titration calorimetric (ITC), computational methods and biochemical approaches. Fluorescence quenching and UV-Visible spectra of the HSA-DCF system confirmed static quenching mechanism and complex formation between HSA and DCF. The thermodynamics results from ITC revealed DCF-HSA interaction was exothermic and spontaneous and involved hydrophobic interactions and hydrogen bonding. The esterase activity of HSA displayed constant Vmax and elevated Km values confirming DCF-HSA competitive interaction. Circular dichroism spectra results revealed structural changes in HSA protein on interaction with DCF. Furthermore, molecular-specific site marker and molecular modelling results affirmed that the binding Site of DCF is Site I of HSA. A significant carbonyl content level in DCF-HSA system suggested protein structure damage. This work is likely to add a better understanding of DCF toxicity in human health and helpful in fortifying the check on food safety.

Bioconcentration of pesticides in zebrafish eleutheroembryos (Danio rerio).

The feasibility of a bioaccumulation test based on the use of zebrafish eleutheroembryos as an alternative to adult-individual-based approaches for REACH application has been evaluated for three test compounds, chlorpyrifos, dicofol and atrazine. Following the OECD 305 guidelines, zebrafish eleutheroembryos (72 h after hatching, hpf) were separately exposed to the investigated pesticides at two nominal concentrations below 1% of its corresponding LC(50). The uptake experiments lasted for 48 h. Then, the exposure medium was replaced by a non-contaminated medium for depuration experiments (up to 72 h). Zebrafish eleutheroembryos (larvae 144 hpf, i.e. at the end of the depuration step) and their corresponding exposure media was sampled at ten different times during each experiment and the concentration of the investigated pesticide determined in both the organisms and in the exposure medium. The experimentally determined pesticide accumulation profiles in the eleutheroembryos demonstrated that atrazine has a very fast accumulation kinetic, reaching steady sate (SS) within 24h. Chlorpyrifos and dicofol did not reach the SS within the 48-h uptake experiments although they exhibit higher accumulations than the former pesticide. Two toxicokinetic models were used to calculate the bioconcentration factor (BCF) of the studied pesticide in zebrafish eleutheroembryos. In the former, the BCF was calculated under SS conditions (BCF(SS)). The second was used when the compounds did not reach the SS during the uptake experiment (BCF(k)). Log BCF values of 3.55 and 3.84 for chlorpyrifos; 0.6 and 1.17 for atrazine, and 3.90 for dicofol were experimentally calculated at selected exposure concentrations. These values have been compared with those reported in related bioaccumulation studies and official databases.

Dicofol residues in eggs and carcasses of captive American kestrels.

American kestrels (Falco sparverius) were fed diets containing 0 (control), 1, 3, 10, and 30 microgram/g (wet wt) of Kelthane. Residues of dicofol and its metabolites were then analyzed in the eggs and carcasses of females. Significant differences occurred among treatments for residues of both p,p'-dicofol and p,p'-dechlorodicofol (DCD) in both eggs and carcasses and for p,p'-dicholorbenzophenone (DCBP) in eggs. Residue concentrations increased with increasing treatment exposure. Residues of p,p'-dicofol, p,p'-DCD, and p,p'-DCBP in eggs were significantly correlated with eggshell quality parameters. Significant correlations also occurred among contaminants in eggs and for individual contaminants between eggs and carcasses. The lowest-observed-dietary-effect concentration for eggshell thinning was 3 microgram/g, whereas 1 microgram/g may be considered to be near a no-observable-adverse-effect concentration. Concentrations of dicofol in potential prey items and eggs of wild birds generally have been lower than dietary-effect concentrations or concentrations in tissues or eggs associated with eggshell thinning and reduced reproductive success.

Mobility of the organochlorine compound dicofol in soil promoted by Pseudomonas fluorescens.

The genetic modified Pseudomonas fluorescens Br 12, resistant to kanamycin and rifampycin, was used to follow the cotransport of the organochlorine acaricide dicofol through a nonsterilized soil column. P. fluorescens was found to bioaccumulate dicofol with the highest bioconcentration factor of 279 within 30 min. Separate soil column experiments where applied P. fluorescens or [14C]dicofol were submitted to heavy rain simulation did not reveal any correlation between the distribution patterns of P. fluorescens and [14C]dicofol in the leachate fractions (r = 0.3). Similar experiments with P. fluorescens that previously had bioaccumulated [14C]dicofol demonstrated a high correlation of these bacteria and radioactivity in the leachate fractions (r = 0.8). The total recovery of radioactivity in the leachate, when [14C]dicofol was previously bioaccumulated in bacteria, was more than two times higher (4.5%) than the total recovery of radioactivity in the leachate when [14C] dicofol was directly applied in the soil (2%). This indicates cotransport by Pseudomonas. Fractionation and analysis of soil columns indicated that most of the bioaccumulated dicofol was rapidly released and adsorbed in soil, while bacteria moved down by leaching.

Kelthane residues in tissues of the tropical penaeid prawn, Metapenaeus monoceros (Fabricius), under sublethal chronic exposure--a monitoring study.

Monitoring the biodegradation and bioaccumulation of kelthane residues were undertaken during reclamation through oxygen consumption (respiration) and target enzyme activity assays. Even after 30 days of reclamation, non-recovery in respiration and target enzyme assays revealed the accumulation of kelthane as a residue in tissues. Thin-layer chromatography demonstrated the presence of kelthane residues as spots in tissue extracts of midgut gland and muscle after a 30-day recovery period. This confirms that kelthane is a persistent insecticide of low biodegradability and capable of accumulating in tissues as residues even at sublethal concentration under chronic exposure.