Assessment of genetic effects and pesticide exposure of farmers in NW Greece.

The present study aims at evaluating potential genotoxic and cytotoxic effects caused by the occupational exposure of farmers to pesticide mixtures in the Aitoloakarnania Prefecture (Greece). The aforementioned assessment was conducted through in vivo Cytokinesis Block Micronucleus assay (CBMN assay) in peripheral blood lymphocytes, in relation to chemical analysis of pesticide residues in blood samples. The exposure of the farmers' population studied to different combinations of pesticides induced significant differences in the frequencies of micronuclei (MN) compared to those of the control group. Furthermore, our results indicated a possible clastogenic and aneugenic effect of pesticides on the genetic material of the farmers exposed. Five pesticides (trifluralin, chlorpyriphos methyl, metolachlor, fenthion and dimethoate) and three metabolites (fenthion sulfone, fenthion sulfoxide and 4,4' DDE) were detected in the 62.5% of blood samples, with mean concentrations ranging from 0.4 ng/ml to 48 ng/ml. Since the farmers studied probably exhibit detectable levels of systematic exposure to the pesticides applied, continuous educational programs focused on the rational and safe use of pesticides, together with implementation of risk communication strategies among farmers are highly recommended.

Photocatalytic degradation kinetics, mechanism and ecotoxicity assessment of tramadol metabolites in aqueous TiO2 suspensions.

This study investigated for the first time the photocatalytic degradation of three well-known transformation products (TPs) of pharmaceutical Tramadol, N-desmethyl-(N-DES), N,N-bidesmethyl (N,N-Bi-DES) and N-oxide-tramadol (N-OX-TRA) in two different aquatic matrices, ultrapure water and secondary treated wastewater, with high (10 mg L(-1)) and low (50 µg L(-1)) initial concentrations, respectively. Total disappearance of the parent compounds was attained in all experiments. For initial concentration of 10 mg L(-1), the target compounds were degraded within 30-40 min and a mineralization degree of more than 80% was achieved after 240 min of irradiation, while the contained organic nitrogen was released mainly as NH4(+) for N-DES, N,N-Bi-DES and NO3(-) for N-OX-TRA. The degradation rates of all the studied compounds were considerably decreased in the wastewater due to the presence of inorganic and organic constituents typically found in effluents and environmental matrices which may act as scavengers of the HO(•). The effect of pH (4, 6.7, 10) in the degradation rates was studied and for N-DES-TRA and N,N-Bi-DES-TRA, the optimum pH value was 6.7. In contrast, N-OX-TRA showed an increasing trend in the photocatalytic degradation kinetic in alkaline solutions (pH 10). The major transformation products were identified by high resolution accurate mass spectrometry coupled with liquid chromatography (HR-LC-MS). Scavenging experiments indicated for all studied compounds the important role of HO(•) in the photocatalytic degradation pathways that included mainly hydroxylation and further oxidation of the parent compounds. In addition, Microtox bioassay (Vibrio fischeri) was employed for evaluating the ecotoxicity of photocatalytically treated solutions. Results clearly demonstrate the progressive decrease of the toxicity and the efficiency of the photocatalytic process in the detoxification of the irradiated solutions.

Environmental fate of the insecticide cypermethrin applied as microgranular and emulsifiable concentrate formulations in sunflower cultivated field plots.

A field dissipation and transport study of the insecticide cypermethrin applied as microgranular (MG) and emulsifiable concentrate (EC) formulations has been conducted in field sunflower cultivations and bare soil plots with two different slopes (1% and 5%). The dissipation of insecticide in soil (on planting rows) was monitored for a period of 193 days. Cypermethrin residual concentrations in the upper soil layer (0-10 cm), 2 days after soil application (DASA), ranged from 0.53 to 0.73 µg g(- 1) when the maximum values were observed 7 DASA, ranged from 1.06 to 1.23 µg g(-1). The dissipation rate was better described by first-order kinetics. The average half-life in cultivated (tilled and planted) plots was 23.07 and 24.24 days for soil slopes 5% and 1%, respectively. In uncultivated (tilled but not planted) plots the respective values were 22.01 and 22.37 days. The insecticide was found below the 10 cm soil layer occasionally in few samples at low concentrations (< 0.02 µg g(- 1)). In runoff water it was detected once (7 days after foliar application, at levels below LOQ), when in sediment it was detectable for seven samplings. The maximum values were observed 7 days after foliar application, when they reached 0.097 and 0.143 µg g(-1) in cultivated plots with soil slopes 1% and 5%; and 0.394 and 0.500 µg g(-1) in uncultivated plots, respectively. The amount of cypermethrin which was transferred by the sediment remained at low levels (less than 0.01% of the totally applied active ingredient), even in plots with 5% inclination. The insecticide was detected in leaves and stems of the sunflower plants after the foliar application up to the day of harvest. On the contrary, in roots it was detectable during the whole cultivation period. No residues were detected in flowers or seeds.

Persistence of oxyfluorfen in soil, runoff water, sediment and plants of a sunflower cultivation.

A field dissipation and transport study of oxyfluorfen in a sunflower cultivation under Mediterranean conditions have been conducted in silty clay plots (cultivated and uncultivated) with two surface slopes (1% and 5%). The soil dissipation and transport of oxyfluorfen in runoff water and sediment, as well as the uptake by sunflower plants, were investigated over a period of 191 days. Among different kinetic models assayed, soil dissipation rate of oxyfluorfen was better described by first-order kinetics. The average half-life was 45 and 45.5 days in cultivated plots with soil slopes 5% and 1% respectively, and 50.9 and 52.9 days in uncultivated plots with soil slopes 5% and 1%. The herbicide was detected below the 10 cm soil layer 45 days after application (DAA). Limited amounts of oxyfluorfen were moved with runoff water and the cumulative losses from tilled and untilled plots with slope 5% were estimated at 0.007% and 0.005% of the initial applied active ingredient, while for the plots with slope of 1%, the respective values were 0.002% and 0.001%. The maximum concentration of oxyfluorfen in sediment ranged from 1.46 µg g(-1) in cultivated plot with soil slope 1% to 2.33 µg g(-1) in uncultivated plot with soil slope 5%. The cumulative losses from tilled and untilled plots with slope 5% were estimated at 0.217% and 0.170% while for the plots with slope of 1%, the respective values were 0.055% and 0.025%. Oxyfluorfen was detected in sunflower plants until the day of harvest; maximum concentrations in stems and leaves (0.042 µg g(-1)) were observed 33 DAA and in roots (0.025 µg g(-1)) 36 DAA. In conclusion, oxyfluorfen hardly moves into silty clay soil and exhibited low run-off potential so it represents a low risk herbicide for the contamination of ground and adjacent water resources.

Pesticide inputs from the sewage treatment plant of Agrinio to River Acheloos, western Greece: occurrence and removal.

This work reports the occurrence and the removal of a wide spectrum of pesticides in municipal wastewaters from the Agrinio region, Greece. Analytical determination was carried out by means of solid-phase extraction followed by gas chromatography equipped with flame thermionic and mass spectrometric detection. The herbicides atrazine, alachlor, isoproturon, and s-metolachlor and the insecticides diazinon, methidathion, fenthion and chlorpyriphos, were more frequently detected in influent and effluent samples while they were also detected in river samples. Isoproturon and diazinon showed the highest concentrations in influent samples that reached concentrations up to 2,328 and 1,486 ng L(-1), respectively. Secondary treatment and cumulative removal rates for herbicides and insecticides ranged between 23-91%, 21->99% and 36->99%, 38->99%, respectively. The data demonstrate that there are significant levels of pesticides entering river waterways.

Occurrence and removal of fungicides in municipal sewage treatment plant.

This work reports the occurrence and the removal of widely used fungicides, in municipal sewage treatment plant receiving combined sewage discharges and operating with primary (mechanical), secondary (activated sludge) and tertiary (sand filtration and chlorination) treatments that are commonly applied in Greece. Target analytes included compounds belonging to triazoles and anilino-pyrimidines. Analytical determination in wastewaters and river water samples was carried out by means of solid-phase extraction followed by gas chromatography equipped with flame thermionic and mass spectrometric detection. The fungicides tebuconazole and cyproconazole were more frequently detected in influent and effluent samples at concentrations up to 1893 and 1735 ng L(-1), respectively. Effluent concentrations were below 691.1 ng L(-1) recorded for tebuconazole. With the exception of triadimefon, all the azole fungicides and pyrimethanil showed relatively low removal efficiencies after secondary and tertiary treatments. Mean removal rates of the fungicides after secondary treatment ranged between 31% for pyrimethanil and 65% for triadimefon. Mean overall removal efficiencies after tertiary treatment ranged between 46% for pyrimethanil and 93% for triadimefon. The findings demonstrate that significant levels of fungicides enter river waterways and that only some of these compounds are being reduced in low levels by municipal wastewater treatment processes.

Organophosphorus pesticide residues in Greek virgin olive oil: levels, dietary intake and risk assessment.

The occurrence of organophosphorus pesticide (OPs) and their metabolite residues was investigated in 167 samples of Greek virgin olive oil during a 2-year (2004-2005) sampling campaign. A total of 30.5% of samples contained detectable residues, although only one sample contained dimethoate residues above the maximum residue limit. Among the seven detected OPs, fenthion and fenthion sulfoxide residues were detected in 10.8 and 14.4% of the samples, respectively, at levels of 0.003-0.61 mg kg(-1). Dimethoate was detected in 10.2% of the samples at 0.003-0.057 mg kg(-1). The acute dietary risk assessment was undertaken by determining the national estimated short-term intake (NESTI); for chronic dietary risk assessment, the national theoretical maximum daily intake (NTMDI) and national estimated daily intake (NEDI) were calculated. The estimated intakes (NESTI and NEDI) of each pesticide were <7 and <0.86% of the corresponding acute reference doses (ARfDs) and acceptable daily intakes (ADIs), respectively. A cumulative risk assessment was performed using the hazard index (HI) and toxicity equivalence factor (TEF), taking into account that OPs share the same toxicological mechanism. The determined HI and TEF values were found to represent only a small portion of the respective ADIs or ARfDs. These results indicate that there is neither acute nor chronic risk for the Greek population through olive oil consumption.

Persistent organochlorine contaminants in liver and fat of birds of prey from Greece.

The concentrations of persistent organic pollutants, such as DDT and its metabolites (DDTs), hexachlorocyclohexane isomers (HCHs), cyclodienes (Cycls), and polychlorinated biphenyls (PCBs), were measured in livers and subcutaneous fat tissues of six Accipitridae and four Falconidae bird species from different areas in Greece. This is the first report of persistent organochlorine (OC) pollutants in birds of prey tissues presented for Greece and the Eastern Mediterranean region. Accumulation patterns of OCs found in birds suggested that the predominant contaminants were p,p'-dichlorodiphenyldichloro-1,1-ethylene (DDE) and PCBs, whereas Cycls and HCHs occurred at low concentrations only. Concentration values of p,p'-DDE ranged from nondetected to 19,518.72 ng/g wet wt in livers and from nondetected to 2679.19 ng/g wet wt in fat. Total PCB levels ranged from 1.01 to 7419.43 ng/g and from 3.25 to 490.10 ng/g wet weight for liver and fat samples, respectively. Higher-chlorinated PCBs such as 118, 138, 153, and 180 predominated in both the liver and subcutaneous fat samples, a pattern comparable to that observed in birds from other European countries. No significant differences in mean concentrations of OCs are detected between species. Hepatic concentrations were in general higher than the fat concentrations showing depleted fat stores in most birds. Concentration ranges were also found in lower or similar levels to those reported for birds in other regions. Variation of OCs levels in bird tissues could be due to different causes of death, with a subsequent effect on body lipid levels, and different feeding and migration habits. The liver PCB levels reported in this study are below the concentrations currently believed to exert mortality or ecotoxicological effects. On the contrary, in some cases p,p'-DDE concentrations were higher than the reported effect values for birds of the same families and could be associated with sublethal effects.

Accumulation profiles of persistent organochlorines in liver and fat tissues of various waterbird species from Greece.

Waterbirds are particularly subject to accumulation of persistent organic pollutants (POPs) that have been shown to constitute a major hazard for this group of birds. Liver and fat tissue from ten species belonging to the orders Ciconiformes (Ardeidae, Ciconiidae, Phoenicopteridae) and Pelicaniformes (Pelecanidae, Phalacrocoracidae) were used as bioindicators in order to assess environmental pollution by POPs (HCHs, DDTs, cyclodienes, PCBs) in Greek wetlands. To our knowledge, this is the first study on POPs in livers of water birds in Greece and Eastern Mediterranean area. The DDTs consisted mainly of p,p'-DDE with percentages over 60% in the great majority of the samples. The highest summation SigmaDDT concentrations were measured in the liver and subcutaneous fat of Phoenicopterus rubber and in Ardea purpurea liver (15565, 24706 and 10406 ng g(-1) wet weight, respectively). Low concentrations of cyclodienes (Cycls) and HCHs were detected occasionally and the contamination pattern of OCPs in most species of waterbirds followed the order summation SigmaDDTs> summation SigmaCycls> summation SigmaHCHs. Individual values of total PCBs reached the levels of 4468 and 3252 ng g(-1) wet weight, for Nycticorax nycticorax and Egretta garzetta samples respectively. Some of the recorded differences in organochlorine concentrations could be due to different causes of death, with a subsequent effect on body lipid levels. Organochlorine pesticides and PCBs residues were lower than those commonly associated with mortality and reduced reproductive success in most species. However, low level exposure to these contaminants may constitute one of the many stressors that in combination could adversely affect bird populations.

Screening method for organophosphorus insecticides and their metabolites in olive oil samples based on headspace solid-phase microextraction coupled with gas chromatography.

This work is focused on the effectiveness of headspace solid-phase microextraction (HS-SPME) for the analysis of nine organophosphorus (OPs) insecticides (dimethoate, diazinon, fenitrothion, malathion, fenthion, parathion ethyl, methyl bromophos, methidathion, ethion) and four metabolites (omethoate, malaoxon, fenthion sulfoxide and fenthion sulfone) residues in olive oil samples. The efficiency of six fibre types with different film thickness was compared. PDMS (100 microm) was found to be the most suitable fibre for the analysis of OPs in olive oils. Optimization of SPME conditions (stirring rate, extraction time, temperature, salt addition) was based on previous developed method in the laboratory that was enriched with additional analytes including major metabolites. In addition, the effect of the oil matrix on the pesticide recoveries was evaluated using spiked oil samples of different composition (acidity, fatty acids, triglycerides, sterols). It was found that only acidity and total amount of sterols are the main factors influencing the SPME efficiency. Matrix effects were compensated for, by using the internal standard method for the quantification of pesticides. The recoveries at three spiking levels were between 80% and 106% with R.S.D. (%) values below 10% in most cases. Good linearity (R2>0.985) was observed in the 0.025-0.50 mg kg(-1) concentration range with satisfactory R.S.D. (%) values of 4.5-10.4%. The method allowed detection of the tested compounds at concentrations below 0.010 mg kg(-1) with GC-FTD detection. In addition, intra- and inter-day precision was satisfactory (R.S.D. (%) <10%). The performance results confirm the usefulness of the proposed methodology for the analysis of OPs in olive oils. Moreover, the maximum residue limits required by European and international regulations can be attained without difficulty. Finally, the method was applied to 30 virgin olive oil samples from major olive production areas of Greece in the framework of an extended monitoring survey of OPs residues in olive oil. The most commonly found pesticides were fenthion, dimethoate and ethion in levels that did not exceed the MRLs.

Monitoring of pesticide residues and their metabolites in surface and underground waters of Imathia (N. Greece) by means of solid-phase extraction disks and gas chromatography.

Seasonal variations of pesticide residues in surface waters and ground waters of the Imathia area of Central Mecedonia (N. Greece) were determined for the period from May 1996 to April 1997. The sampling cruises included eight sites in rivers Aliakmon, Loudias, Tripotamos, Arapitsa and Canal-66, seven water springs in the mountain Vermion, seven rainfall water collection stations and one hundred underground points. Solid-phase extraction disks followed by gas chromatographic techniques with flame thermionic detection, electron capture detection and mass-selective detection were used for the monitoring of various pesticides their transformation products in environmental waters. The most commonly encountered pesticides in underground waters, were alachlor, atrazine, desethylatrazine (DEA), metolachlor, molinate, propanil, simazine, carbofuran, diazinon and parathion methyl. The above compounds including propazine, trifluralin, malathion, parathion ethyl, lindane, alpha-benzene hexachloride (alpha-BHC), beta-BHC, 4,4'-DDE and heptachlor were determined in river waters. The higher concentrations in underground waters were measured during the period from May to August, 1996, following seasonal application and diminished significantly during the autumn and winter. Water pollution by triazine and chloroacetanilides was highest in the estuarine areas; showing that many of these compounds are transported significant distances from their application sites. The major inputs of atrazine, alachlor, simazine and metolachlor occurred in May and June just after their application. Atrazine, DEA, diazinon and metolachlor were also detected in spring waters at concentration levels below 0.006 microgram/l. Finally, atrazine, DEA, carbofuran, simazine, diazinon, parathion ethyl and parathion methyl were detected in rainfall water samples collected in the agricultural area of Imathia (central part of the plain).

Concentration and bioaccumulation of organochlorine pesticide residues in herons and their prey in wetlands of Thermaikos Gulf, Macedonia, Greece.

Concentrations of the principal organochlorine insecticides were determined in eggs and freshly dead chicks of the Squacco heron (Ardeola ralloides), Little Egret (Egretta garzetta) and Night Heron (Nycticorax nycticorax), as well as in frogs (Rana sp.), the main heron prey. Material was collected from the wetlands of the Thermaikos Gulf (Macedonia, northern Greece) in 1992 and 1993. Residues of the organochlorine pesticides alpha-BHC, beta-BHC, lindane, 4,4'-DDD, 4,4'-DDE, heptachlor and dieldrin were found in the eggs, chicks and prey of the herons. alpha-BHC, beta-BHC, and lindane had highest concentration in the Night Heron and lowest in the Little Egret. In all samples examined, the bioconcentration factors (BCF) of these compounds had very high values. BCF of pollutants for the eggs of the Squacco Heron were at lower levels than those of its chicks. BCF for frogs were in almost all cases lower than those for the other samples. Biomagnification factor (BMF) for 4,4'-DDE and beta-BHC had the highest values of all other compounds (except in the Night Heron). BMF for the eggs of the Squacco Heron were greater than for its chicks. Variation in the pesticide contents in the different heron species is attributed to different feeding habits; the exception being the occurrence of dieldrin in eggs only and 4,4'-DDE as a remnant of past spraying. Amounts of pesticides detected in this study are too low to affect eggshell thickness in the Squacco Heron or have other effects on the wildlife of the area.