The Impact of Pesticides - Academy Publish

of the study. A face-to-face questionnaire was completed to obtain information on a)standard demographic data (age, gender, etc), b) individual lifestyle (diet, smokinghabit, alcohol and medicine consumption), c) occupational aspects (workinghours/days, years of exposure to pesticides, use of protective measures, etc), and d)pesticides used.The study involved 315 subjects divided into two groups. The first group: Intensivecrop workers (horticultural crops) consisted of 53 pesticide sprayers or pesticideapplicators and 59 agricultural workers or farmers, and the control group consistedof 112 donors, from the same area, with no history of occupational exposure topesticides or any potential genotoxic agent. The second group: Extensive cropworkers (cereals and oil seeds) consisted of 48 pesticide sprayers or pesticideapplicators and a control group of 50 donors.Peripheral blood heparinized samples were obtained from all the subjects involvedin the study.BiomarkersThe variable used to describe pesticide exposure were butyrylcholinesterase (BChE)and acetylcholinesterase (AChE). Other indirect measures of exposure includedutilization of protective gear during mixing/loading or during application ofpesticides. Farmers can be expected to accurately recall details of their use ofpesticides because it is a significant part of their farming operations.AChE activity in erythrocytes: An aliquot of washed erythrocytes were haemolyzedby adding demineralized water at a 1:10 dilution. The hydrolysis rate ofacetylthiocholine iodide (substrate) in erythrocyte dilution was measured at 405 nmwith spectrophotometer by the reaction with DTNB to give the yellow 5-thio-2-nitrobenzoate anion. Enzyme activity was expressed as U/L of Red Blood Cells(RBC) (Ellman et al., 1961).BChE plasmatic activity: The hydrolysis rate of butyrilthiocholine (substrate) inplasma was measured at 405 nm with spectrophotometer by the reaction ofthiocholine iodide with DTNB, to give the yellow 5-thio-2-nitrobenzoate anion.Enzyme activity was expressed as U/L. (Ellman et al., 1961.)Biomarkers of effect concern an assessment of early or late adverse effects of achemical or another factor on a physiological system, organ or organism. Theprimary purpose of using biomarkers of effect is surveillance, that is theidentification of individuals or a population at risk to adverse health effects so thatpreventive measures can be taken. Although a biomarker of effect is usually alsorelated to exposure to a specific chemical, it is generally more closely related to theoccurrence of an adverse health effect (De Zwart, 1999). Pesticides are example ofagents that act as pro-oxidants and elicit effects in multiple organs. In some cases,these prooxidant effects occur alongside pesticide-induced alterations in targetenzymes (Limón-Pacheco and Gonsebatt, 2009). Besides, it was verified an increaseof thiobarbituric acid reactive species (TBARS). To provide concise guidance fordetecting DNA damage (strand breaks, alkali labile sites, crosslinking andincomplete excision repair sites) the single cell gel or Comet assay is used in cellssampled from individuals potentially exposed to genotoxic carcinogens (Albertini etal, 2000).Catalase activity in erythrocytes (CAT): Erythrocytes were haemolyzed by addingice-cold demineralized ultrapure water at a 1:100 dilution. CAT activity inhaemolysate erythrocytes was measured by monitoring the decrease in H 2 O 2concentration spectrophotometrically over time (Aebi, 1984). The specific activityof each sample was calculated on the basis that one unit of enzyme activity wasdefined as the activity required to degrades 1 mole hydrogen peroxide during 60 s/gHb.AcademyPublish.org - The Impact of Pesticides86