Occurrence of Banned and Currently Used Herbicides, in Groundwater of Northern Greece: A Human Health Risk Assessment Approach.

The presence of pesticide residues in groundwater, many years after their phase out in European Union verifies that the persistence in aquifer is much higher than in other environmental compartments. Currently used and banned pesticides were monitored in Northern Greece aquifers and a human health risk assessment was conducted. The target compounds were the herbicides metolachlor (MET), terbuthylazine (TER), atrazine (ATR) and its metabolites deisopropylatrazine (DIA), deethylatrazine (DEA) and hydroxyatrazine (HA). Eleven sampling sites were selected to have representatives of different types of wells. Pesticides were extracted by solid-phase extraction and analyzed by liquid chromatography. MET was detected in 100% of water samples followed by ATR (96.4%), DEA and HA (88.6%), DIA (78.2%) and TER (67.5%). ATR, DIA, DEA, HA, MET and TER mean concentrations detected were 0.18, 0.29, 0.14, 0.09, 0.16 and 0.15 µg/L, respectively. Obtained results were compared with historical data from previous monitoring studies and temporal trends were assessed. Preferential flow was the major factor facilitating pesticide leaching within the month of herbicide application. Moreover, apparent age of groundwater and the reduced pesticide dissipation rates on aquifers resulted of long-term detection of legacy pesticides. Although atrazine had been banned more than 18 years ago, it was detected frequently and their concentrations in some cases were over the maximum permissible limit. Furthermore, human health risk assessment of pesticides was calculated for two different age groups though drinking water consumption. In all examined wells, the sum of the HQ values were lower than the unity. As a result, the analyzed drinking water wells are considered safe according to the acute risk assessment process. However, the presence of atrazine residues causes concerns related with chronic toxicity, since ATR R values were greater than the parametric one of 1 × 10-6 advised by USEPA, for both age groups.

Fluopyram removal from agricultural equipment rinsing water using HSF pilot-scale constructed wetlands.

Fluopyram is a novel broad-spectrum fungicide with nematocidal activity, and as an extensively used pesticide, it could cause toxicity in nontarget organisms. The aim of this study was to explore the efficiency of five horizontal subsurface flow (HSF) constructed wetlands (CWs) to remove fluopyram from rinsing water produced during the cleaning of pesticide spraying equipment. Four CWs, namely WG-R, WG-R-P, WG-C, and WG-U, contained fine gravel as porous media. WG-R and WG-R-P were planted with Phragmites australis, WG-C with Typha latifolia, and WG-U was left unplanted. Bioaugmentation with plant growth-promoting rhizobacteria was conducted in WG-R-P unit. The fifth unit (WGZ-R) planted with Phragmites australis and contained gravel and zeolite as porous media. All of CWs were loaded on a daily basis from December 2019 to January 2021 with water fortified with fluopyram. The removal rate follows the pattern of WG-R-P (70.67%) > WGZ-R (62.06%) > WG-C (59.98%) > WG-R (36.10%) > WG-U (25.09%). The most important parameters affecting the fluopyram removal were bioaugmentation, zeolite presence in porous media, and plant species. The WG-R-P unit showed higher fluopyram removal in comparison to the WG-R (increase about 96%), the zeolite increased the fluopyram removal by 72%, and the WG-C unit showed 66% higher fluopyram removal than the WG-R unit.

Pesticide prioritization approaches and limitations in environmental monitoring studies: From Europe to Latin America and the Caribbean.

Assessment and management of issues related to pesticide residues, such as environmental fate, monitoring and toxicity, are complex and, in many cases, require costly studies. The early establishment of a priority list of pesticides that should be monitored and assigned to a restricted-use policy is an important issue of post-registration Risk Assessment (RA). Various pesticide registration approaches have been adopted by different countries with those from Europe and the USA being the most popular, constituting the major prototypes for registration approaches in other countries. Adoption of pesticide registration and monitoring systems developed in Europe or USA by Latin American and Caribbean countries may underestimate factors affecting the environmental fate and toxicity of pesticides in their own countries. Incentive for this short review was the activities undertaken during the three KNOWPEC workshops held in Costa Rica, Argentina and Bolivia where European pesticide experts met Latin American experience in the form of Costa Rica's exceptional environmental conditions and ecology, Argentina's and Uruguay's soyisation and Bolivia's contrasting climate and agricultural zones. During the parallel activities of the workshop - including scientific presentations, field trips, interviews and meetings among European partners and pesticide stakeholders in Latin America, - the whole pesticide chain (import-export, trade, application, plant protection-efficacy, residues, monitoring, remediation and risk) was studied and clarified. Recently-published chemical prioritization studies were reviewed to consider their use as a tool to support risk assessments. Differences in regional practices are highlighted as regards to the establishment of RA or prioritization strategy in European and Latin American regimes. General guidance of establishing a cost-effective pesticide monitoring scheme in water bodies of Latin America and the Caribbean (LAC) is also proposed. Moreover, we summarize the most important factors that should be taken into consideration for prioritization approaches and categorization used in pesticide environmental monitoring studies. Consideration of current RA approaches and limitations, and pesticide prioritization exercises highlighted in this Commentary could assist in the management of pesticides in Latin America and Caribbean.

Assessment and management of pesticide pollution at a river basin level part I: Aquatic ecotoxicological quality indices.

During the last decade numerous monitoring programs have been conducted in order to assess pesticide pollution in catchments. This effort has led to the production of large, complex data sets of environmental results making the task of evaluation of the aquatic chemical status more difficult. Furthermore, the evaluation of the chemical status of the water ecosystems is one of the main aspects which should be considered in a River Basin Management Plan. In this study, two indices were developed in order to assess the combined pesticide ecotoxicity to aquatic non-target organisms, the Aquatic Quality Index of Short term Toxicity of Pesticides (AQI ShToxP) and the Aquatic Quality Index of Long term Toxicity of Pesticides (AQI LToxP). These indices were applied to the environmental results obtained from an intensive monitoring study of 302 pesticides in 102 stationary sampling stations located on the surface aquatic network of the Pinios River Basin, in Greece, in 2011 and 2012. The evaluation of the surface water quality was achieved by taking into consideration the frequency and the intensity of exposure of the aquatic organisms to pesticides above the respective ecotoxicological quality objectives such as the acute or chronic term predicted no-effect concentrations derived from risk assessment. Seventy-five pesticides, that have been previously identified as the River Basin Specific Pollutants of Pinios by an environmental and human risk hierarchy exercise, were assessed. It appears, from the implementation of the two indices, that the detected pesticides in the surface aquatic ecosystem of the Pinios River Basin exert significant pressure on the aquatic non-target organisms especially at the chronic effect level. The developed AQI ShToxP and AQI LToxP indices, as well as the proposed quality classification system could be valuable communication and interpretation tools for River Basin Management Plans that can contribute in the restoration of environmental health.

Assessment and management of pesticide pollution at a river basin level part II: Optimization of pesticide monitoring networks on surface aquatic ecosystems by data analysis methods.

The high cost of extensive pesticide monitoring studies, required for the protection of water resources, and the necessity of early identification of environmental threats, highlighted the need for prioritization of pesticides and sampling sites to be monitored. The aim of this study was to develop an optimum surface water monitoring network at a catchment scale including only the sites of a catchment vulnerable to pesticide pollution. The identification of sampling sites vulnerable to pesticide pollution (VPS) was based on the data of an intensive monitoring survey of 302 pesticides in 102 stationary sampling sites located on the surface water network of a river basin. In the proposed methodology the left-censored data of the analytical results derived from the above mentioned monitoring campaign were included in the statistical analyses by transforming all the raw data into categorical variables and arranging them in ordinal scales based on ecotoxicological thresholds derived from pesticide toxicity tests on aquatic non-target organisms. The categorized data were subjected to Categorical Principal Component Analysis with Optimal Scaling. For the identification of the VPS, the Squared Mahalanobis Distance criterion was applied on the extracted values (scores) of the significant principal components. With this methodology a 46% reduction in the number of the monitoring stations was achieved. This approach will be valuable in establishing more cost effective monitoring schemes in the future in other basins and in developing targeted measures to eliminate or limit the effect of critical pollution sources in surface aquatic systems. Moreover, by applying the proposed methodology, historical monitoring data can be used to initiate more efficient pesticide monitoring campaigns in the future.

Pesticide monitoring and ecotoxicological risk assessment in surface water bodies and sediments of a tropical agro-ecosystem.

A pesticide monitoring study including 80 and 60 active ingredients (in surface waters and sediments, respectively) was carried out in a river basin in Costa Rica during 2007-2012. A special emphasis was given on the exceptional ecological conditions of the tropical agro-ecosystem and the pesticide application strategies in order to establish a reliable monitoring network. A total of 135 water samples and 129 sediment samples were collected and analyzed. Long-term aquatic ecotoxicological risk assessment based on risk quotient in three trophic levels was conducted. Short-term risk assessment was used to calculate the toxic unit and prioritization of sampling sites was conducted by the sum of toxic units in both aquatic and sediment compartments. Dimethoate (61.2 µg/L), propanil (30.6 µg/L), diuron (22.8 µg/L) and terbutryn (4.8 µg/L) were detected at the highest concentrations in water samples. Carbendazim and endosulfan were the most frequently detected pesticides in water and sediment samples, respectively. Triazophos (491 µg/kg), cypermethrin (71.5 µg/kg), permethrin (47.8 µg/kg), terbutryn (38.7 µg/kg), chlorpyrifos (18.2 µg/kg) and diuron (11.75 µg/kg) were detected at the highest concentrations in sediment samples. The pesticides carbendazim, diuron, endosulfan, epoxyconazole, propanil, triazophos and terbutryn showed non-acceptable risk even when a conservative scenario was considered. Sum TUsite higher than 1 was found for one and two sampling sites in water and sediment compartments, respectively, suggesting high acute toxicity for the ecosystem. MAIN FINDING OF THE WORK: Exceptional ecological conditions of the tropical agro-ecosystem affect the fate of pesticides in water and sediment environment differently than the temperate one.

Pesticides in the rivers and streams of two river basins in northern Greece.

The pollution caused by pesticides, and their ecotoxicological implications were investigated in water samples from the Strymonas and Nestos river basins (Northern Greece). Chlorpyrifos was the most frequently detected pesticide in both basins (42 and 37% in the Strymonas and Nestos basins, respectively), followed by fluometuron and terbuthylazine (25 and 12%, Strymonas), and bentazone and boscalid (24 and 10%, Nestos). The Annual Average and the Maximum Allowable Concentration of Environmental Quality Standards set in European Union Directives were exceeded in several cases by alphamethrin and chlorpyrifos. Risk Quotient assessment revealed significant ecological risk towards the aquatic organisms in over 20% of the water samples. Insecticides (mostly pyrethroids and organophosphosphates) contributed more in the ecotoxicological risk than herbicides and fungicides. The three main rivers in the current study (Strymonas, Aggitis, Nestos) exhibited similar sum of RQs indicating that aquatic life in all three of them was at the same risk level. However, the sums of RQs were higher in the various streams monitored than the three rivers.

The Plant as Metaorganism and Research on Next-Generation Systemic Pesticides - Prospects and Challenges.

Systemic pesticides (SPs) are usually recommended for soil treatments and as seed coating agents and are taken up from the soil by involving various plant-mediated processes, physiological, and morphological attributes of the root systems. Microscopic insights and next-generation sequencing combined with bioinformatics allow us now to identify new functions and interactions of plant-associated bacteria and perceive plants as meta-organisms. Host symbiotic, rhizo-epiphytic, endophytic microorganisms and their functions on plants have not been studied yet in accordance with uptake, tanslocation and action of pesticides. Root tips exudates mediated by rhizobacteria could modify the uptake of specific pesticides while bacterial ligands and enzymes can affect metabolism and fate of pesticide within plant. Over expression of specific proteins in cell membrane can also modify pesticide influx in roots. Moreover, proteins and other membrane compartments are usually involved in pesticide modes of action and resistance development. In this article it is discussed what is known of the physiological attributes including apoplastic, symplastic, and trans-membrane transport of SPs in accordance with the intercommunication dictated by plant-microbe, cell to cell and intracellular signaling. Prospects and challenges for uptake, translocation, storage, exudation, metabolism, and action of SPs are given through the prism of new insights of plant microbiome. Interactions of soil applied pesticides with physiological processes, plant root exudates and plant microbiome are summarized to scrutinize challenges for the next-generation pesticides.

Environmental and human risk hierarchy of pesticides: A prioritization method, based on monitoring, hazard assessment and environmental fate.

A pesticide prioritization approach was developed and implemented in the Pinios River Basin of Central Greece. It takes under consideration the Level of Environmental Risk containing information on the frequency of occurrence of pesticides above environmental thresholds, the intensity of this occurrence and the spatial distribution as well as information about the fate and behavior of pesticides in the environment and the potential to have adverse impact on humans' health. Original 3-year monitoring data from 102 Stationary Sampling Sites located on rivers and their tributaries, reservoirs, streams and irrigation/drainage canals giving rise to a collection of 2382 water samples resulting in 7088 data sets, were included in this integrated prioritization study. Among 302 monitored active ingredients, 119 were detected at least once and the concentrations found in the aquatic systems for 41% of compounds were higher than the respective lowest Predicted Non-Effect Concentration (PNEC) values. Sixteen and 5 pesticides were found with risk ratios (MECmax/PNEC) above 10 (high concern) and 100 (very high concern), respectively. However, pesticides with maximum Measured Environmental Concentration (MECmax) values exceeding by 1000 times the respective lowest PNEC values were also found which were considered of extremely high concern; in the latter group were included prometryn, chlorpyrifos, diazinon, λ-cyhalothrin, cypermethrin, α-cypermethrin deltamethrin, ethalfluralin and phosmet. The sensitivity of the analytical methods used in the monitoring study was considered inadequate to meet the toxicological endpoints for 32 pesticides. The widest distribution of occurrence in the Stationary Sampling Sites of the monitoring program was found for the pesticides, prometryn, fluometuron, terbuthylazine, S-metolachlor, chlorpyrifos, diphenylamine, acetochlor, alachlor, 2,4-D, etridiazole, imidacloprid and lindane (γ-ΗCH). Among the 27 priority pesticides included in the Directive 2013/39/EU, in the present study 13 pesticides were considered as candidates for River Basin Specific Pollutants (RBSP) for the River Basin of Pinios. Among the 30 pesticide specific pollutants that were established by the Greek authorities, in the present study only 6 were considered as candidates for RBSP for the river basin of Pinios. As a result of the implementation of the prioritization approach developed in this study a total of 71 pesticides were identified as being RBSP for the river basin of Pinios. The higher Level of Environmental Risk was found to be exerted by the organophosphorus insecticide chlorpyrifos, followed by lindane and prometryn. The present study provides background information for important decisions to be made concerning the selection of pesticides which should be included in the target analyte list of new monitoring and screening programs of surface water quality in the Pinios River Basin. In addition, the prioritization approach proposed here can be useful for the development of River Basin Management Plans.

Pesticides in the surface waters of Lake Vistonis Basin, Greece: Occurrence and environmental risk assessment.

A study was undertaken for the evaluation of the pesticide pollution caused by the agricultural activities in the basin of Lake Vistonis, Greece during the years 2010-2012. Water samples were collected from Lake Vistonis, four major rivers and various small streams and agriculture drainage canals. The concentration of 302 compounds was determined after solid-phase extraction of the water samples and subsequent LC-MS/MS and GC-MS/MS analysis of the extracts. Overall, herbicides were the most frequently detected pesticides (57%), followed by insecticides (28%) and fungicides (14%). In Lake Vistonis 11 pesticides were detected. Specifically, fluometuron was detected in the 75% of the samples (maximum concentration 0.088 µg/L) whereas lambda-cyhalothrin was detected in all the samples of spring 2011 and alphamethrin in all the samples of spring 2012 (maximum concentration 0.041 and 0.168 µg/L, respectively). In the rivers and drainage canals 68 pesticides were detected. Specifically, fluometuron was detected in the 53% of the samples (maximum concentration 317.6 µg/L) followed by chlorpyrifos and prometryn (16 and 13% of the samples respectively). An environmental risk assessment was performed by employing the Risk Quotient (RQ) method. The risk assessment revealed that at least one pesticide concentration led to a RQ>1 in 20% of the samples. In Lake Vistonis, alphamethrin and lambda-cyhalothrin concentrations resulted in RQ>1, whereas in the other water bodies this was mainly the result of chlorpyrifos-methyl and alphamethrin exposure. In contrast, herbicide and fungicide concentrations contributed substantially less to environmental risks.

Determination of mycotoxins in pomegranate fruits and juices using a QuEChERS-based method.

A rapid and accurate analytical method for the determination of three Alternaria mycotoxins (alternariol, alternariol monomethyl ether, and tentoxin) in pomegranate samples (fruits and juices) was developed and validated. The overall average recoveries ranged for 82.0-109.4% and the relative standard deviations were from 1.2% to 10.9%. The optimized and validated method was applied to detect the presence of the target mycotoxins in real samples (fruits and juices) purchased from Greek markets. Mycotoxins were not found in any of the analyzed samples. Also, artificially inoculated pomegranate fruits with six different Alternaria alternata species complex isolates, known to produce the target mycotoxins on pure cultures, were analyzed and alternariol concentrations found ranged from 0.3 to 50.5 µg/g, alternariol monomethyl ether from 0.5 to 32.3 µg/g, while tentoxin was not detected. The developed analytical method can be used for the routine monitoring of the major Alternaria mycotoxins in pomegranates.

Comparison and evaluation of two methods for the pesticide residue analysis of organophosphates in yerba mate

Abstract Microwave Assisted Extraction and a modified CEN-QuEChERS methodology were evaluated as extraction and clean up procedures for the simultaneous analysis of 42 organophosphate pesticides in yerba mate (Ilex paraguaiensis). The obtained extracts were analyzed by gas chromatography using a flame photometric detector. Linearity, recovery percentages, relative standard deviations, detection and quantification limits and matrix effects were determined according to DG-SANCO guidelines for both methods. At 0.2 and 0.5 mg/kg the evaluated methods showed percentages recoveries between 70 and 120% for most of the analytes. Using Microwave Assisted Extraction methodology, 33 pesticide residues could be properly analyzed whereas only 27 could be determined with the proposed modified QuEChERS. All relative standard deviation were below 18% except for omethoate and disulfoton sulfone when evaluated by the modified QuEChERS. The limits of detection in both methodologies were 0.2 mg/kg for most of the analyzed compounds. The average detection limit for QuEChERS was 0.04 mg/kg. For 19 of the analytes determined through Microwave Assisted Extraction the lowest validated level were 0.004 mg/kg. Signal suppression/enhancement was observed for most of the pesticides, thus matrix-matched calibration curves were used for quantification. The Microwave Assisted Extraction and QuEChERS procedures studied could detect the organophosphate pesticides above the MRL fixed for "mate" by the European Union. They have been successfully applied for the determination of organophosphate pesticide residues in commercial samples and the positives were confirmed through GC–(ITD)-MS.

A pesticide monitoring survey in rivers and lakes of northern Greece and its human and ecotoxicological risk assessment.

A pesticide monitoring study covering the main rivers and lakes of Northern Greece (Macedonia, Thrace and Thessaly) was undertaken. A total of 416 samples were collected over a 1.5-year sampling period (September 1999- February 2001) from six rivers and ten lakes. The water samples were analyzed with an off-line solid phase extraction technique coupled with a gas chromatography ion trap mass spectrometer using an analytical method for 147 pesticides and their metabolites, including organochlorines, organophosphates, triazines, chloroacetanilides, pyrethroids, carbamates, phthalimides and other pesticides (herbicides, insecticides and fungicides). Based on the pesticide survey results, a human health carcinogenic and non-carcinogenic risk assessment was conducted for adults and children. Ecotoxicological risk assessment was also conducted using default endpoint values and the risk quotient method. Results showed that the herbicides metolachlor, prometryn, alachlor and molinate, were the most frequently detected pesticides (29%, 12.5%, 12.5% and 10%, respectively). They also exhibited the highest concentration values, often exceeding 1 µg/L. Chlorpyrifos ethyl was the most frequently detected insecticide (7%). Seasonal variations in measured pesticide concentrations were observed in all rivers and lakes. The highest concentrations were recorded during May-June period, right after pesticide application. Concentrations of six pesticides were above the maximum allowable limit of 0.1 µg/L set for drinking water. Alachlor, atrazine and a-HCH showed unacceptable carcinogenic risk estimates (4.5E-06, 4.6E-06 and 1.3E-04, respectively). Annual average concentrations of chlorpyriphos ethyl (0.031 µg L), dicofol (0.01 µg/L), dieldrin (0.02 µg/L) and endosulfan a (0.065 µg/L) exceeded the EU environmental quality standards. The risk quotient estimates for the insecticides chorpyrifos ethyl, diazinon and parathion methyl and herbicide prometryn were above acceptable risk values. The coupling of monitoring data to probabilistic human and ecotoxicological risk estimates could find use by Greek regulatory authorities, proposing effective pollution management schemes.

Effect of specific plant-growth-promoting rhizobacteria (PGPR) on growth and uptake of neonicotinoid insecticide thiamethoxam in corn (Zea mays L.) seedlings.

BACKGROUND: Corn (Zea mays L.) is one of the most important cereal crops in the world and is used for food, feed and energy. Inoculation with plant-growth-promoting rhizobacteria (PGPR) would reduce the use of chemical fertilisers and pesticides and could be suggested as an alternative practice for sustainable production of corn in modern agricultural systems. In this study, the effect of two Bacillus PGPR formulated products, Companion (B. subtilis GB03) and FZB24 (B. subtilis FZB24), on corn growth and root uptake of insecticide thiamethoxam was investigated. RESULTS: All bacterial treatments enhanced root biomass production by 38-65% compared with the uninoculated control, with no stimulatory effect of PGPR on above-ground biomass of corn. The uptake results revealed that, in plants inoculated with the PGPR B. subtilis FZB24 and B. subtilis GB03, singly or in combination, the uptake and/or systemic translocation of thiamethoxam in the above-ground corn parts was significantly higher at the different growth ages compared with the control receiving no bacterial treatment. CONCLUSION: The findings suggest that the PGPR-elicited enhanced uptake of thiamethoxam could lead to improved efficiency of thiamethoxam using reduced rates of pesticides in combination with PGPR as an alternative crop protection technique.

Occurrence of pesticides in transboundary aquifers of North-eastern Greece.

A five-year groundwater monitoring program undertaken in Evros (north-east Greece), showed a diversification in the levels of pesticide residues detected in adjacent transboundary aquifers. During the first two years 37 wells, including irrigation, drinking water and artesian wells were monitored while the next three years the survey was focused on the 11 most contaminated wells. The presence of pesticide residues was also monitored in the phreatic horizon (shallow groundwater) of four experimental boreholes drilled in the respective margins of four fields. Among the compounds found alachlor, metolachlor, atrazine, desethylatrazine (DEA), desisopropylatrazine (DIA) and caffeine were constantly detected. Pesticide concentrations were much lower (up to 1.54 µg/L) in the water of the monitored drinking water wells (deep groundwater aquifers) compared to those found in the phreatic horizon (experimental boreholes) of the respective areas (up to 5.20 µg/L). DEA to atrazine concentration ratios (DAR) determined for the phreatic horizon of the three boreholes and respective wells were lower than 1, indicating that preferential flow was the cause of the fast downward movement of atrazine to the phreatic horizon. In contrast the DAR for the fourth borehole and the adjacent well were greater than 1 indicating the absence of preferential flow of atrazine. Catabolic processes of the soil converted atrazine to DEA which is more mobile than atrazine itself through chromatographic (darcian) flow. This differential behavior of pesticides in adjacent aquifers (3 km) was further investigated by determining the apparent age of water in the two wells. The apparent age of the water present in the first aquifer was 21.7 years whereas the apparent age of that in the second aquifer was approximately 1.2 years. The faster replenishing rate of the latter is an indication that this aquifer is very vulnerable to contamination with pollutants present in the infiltrated soil water.

Biotransformation of atrazine and metolachlor within soil profile and changes in microbial communities.

Biotransformation studies of atrazine, metolachlor and evolution of their metabolites were carried out in soils and subsoils of Northern Greece. Trace atrazine, its metabolites and metolachlor residues were detected in field soil samples 1 year after their application. The biotransformation rates of atrazine were higher in soils and subsoils of field previously exposed to atrazine (maize field sites) than in respective layers of the field margin. The DT(50) values of atrazine ranged from 5 to 18d in the surface layers of the adapted soils. DT(50) values of atrazine increased as the soil depth increased reaching the value of 43 d in the 80-110 cm depth layer of adapted soils. Metolachlor degraded at slower rates than atrazine in surface soils, subsoils of field and field margins with the respective DT(50) values ranging from 56 to 72 d in surface soils and from 165 to 186 d in subsoils. Hydroxyatrazine was the most frequently detected metabolite of atrazine. The maximum concentrations of metolachlor-OXA and metolachlor-ESA were detected in the soil layers of 20-40 cm depth after 90 d of incubation. Principal Component Analysis (PCA) of soil Phospholipid Fatty Acids (PLFAs), fungal/bacterial and Gram-negative/Gram-positive ratios of the PLFA profiles revealed that the higher biotransformation rates of atrazine were simultaneously observed with the abundance of Gram-negative bacteria while the respective rates of metolachlor were observed in soil samples with abundance of fungi.

Evaluation of plant-growth-promoting rhizobacteria, acibenzolar-S-methyl and hymexazol for integrated control of Fusarium crown and root rot on tomato.

BACKGROUND: Plant growth-promoting rhizobacteria (PGPR) can be potential agents for biological control of plant pathogens, while their combined use with conventional pesticides may increase their efficacy and broaden the disease control spectrum. The effect of four different Bacillus sp. PGPR strains (B. subtilis GB03 and FZB24, B. amyloliquefaciens IN937a and B. pumilus SE34) applied individually and in mixtures, as well as in combined use with acibezolar-S-methyl (ASM) and hymexazol, on plant growth promotion and on the control of Fusarium crown and root rot (FCRR) of tomato was evaluated. RESULTS: All PGPR strains promoted the tested plant growth characteristics significantly. A higher promoting effect was provided by SE34. Experiments on population dynamics of PGPR strains revealed that, after 28 days of incubation, populations of strain SE34 remained stable, while the remaining bacterial strains showed a slight decline in their population densities. The GB03 and FZB24 strains provided a higher disease suppression when applied individually. However, application of IN937a in a mixture with GB03 provided a higher control efficacy of Fusarium oxysporum f. sp. radicis-lycopersici (Forl). Treatment of tomato plants with ASM resulted in a small reduction in disease index, while application of hymexazol provided significantly higher control efficacy. Combined applications of the four PGPR strains with either ASM or hymexazol were significantly more effective. CONCLUSION: The results of the study indicate that, when bacilli PGPR strains were combined with pesticides, there was an increased suppression of Forl on tomato plants, and thus they may prove to be important components in FCRR integrated management.

Leaching of Br-, metolachlor, alachlor, atrazine, deethylatrazine and deisopropylatrazine in clayey vadoze zone: a field scale experiment in north-east Greece.

An extensive four-year research program has been carried out to explore and acquire knowledge about the fundamental agricultural practices and processes affecting the mobility and bioavailability of pesticides in soils under semi-arid Mediterranean conditions. Pesticide leaching was studied under field conditions at five different depths using suction cups. Monitoring of metolachlor, alachlor, atrazine, deethylatrazine (DEA), deisopropylatrazine (DIA), and bromide ions in soil water, as well as dye patterns made apparent the significant role of preferential flow to the mobility of the studied compounds. Irrespective to their adsorption capacities and degradation rates, atrazine, metolachlor and bromide ions were simultaneously detected to 160 cm depth. Following 40 mm irrigation, just after their application, both alachlor and atrazine were leached to 160 cm depth within 18 h, giving maximum concentrations of 211 and 199 µg L(-1), respectively. Metolachlor was also detected in all depth when its application was followed by a rainfall event (50 mm) two weeks after its application. The greatest concentrations of atrazine, alachlor and metolachlor in soil water were 1795, 1166 and 845 µg L(-1), respectively. The greatest concentrations of atrazine's degradation products (both DEA and DIA) appeared later in the season compared to the parent compound. Metolachlor exhibited the greatest persistence with concentrations up to 10 µg L(-1) appearing in soil water 18 months after its application. Brilliant blue application followed by 40 mm irrigation clearly depict multi-branching network of preferential flow paths allowing the fast flow of the dye down to 150 cm within 24 h. This network was created by soil cracks caused by shrinking of dry soils, earthworms and plant roots. Chromatographic flow of the stained soil solution was evident only in the upper 10-15 cm of soil.

Biodegradation of soil-applied pesticides by selected strains of plant growth-promoting rhizobacteria (PGPR) and their effects on bacterial growth.

A laboratory study was conducted to investigate the influence of four PGPR strains on the degradation of five soil applied pesticides and their effects on bacterial growth. Interactions of Bacillus subtilis GB03, Bacillus subtilis FZB24, Bacillus amyloliquefaciens IN937a and Bacillus pumilus SE34 with two concentrations of acibenzolar-S-methyl, metribuzin, napropamide, propamocarb hydrochloride and thiamethoxam in liquid culture and soil microcosm were studied. The degradation of acibenzolar-S-methyl by all PGPR tested in low and high concentration, was 5.4 and 5.7 times, respectively, faster than that in non-inoculated liquid culture medium. At the end of the 72-h liquid cultured experiments, 8-18, 9-11, 15-36 and 11-22% of metribuzin, napropamide, propamocarb hydrochloride and thiamethoxam, respectively, had disappeared from PGPR inoculated medium. Under the soil microcosm experimental conditions, the half-lives of acibenzolar-S-methyl incubated in the presence of PGPR strains spiked at 1.0 and 10.0 mg kg(-1) were 10.3-16.4 and 9.2-15.9 days, respectively, markedly lower compared with >34.2 days in the control. From the rest pesticides studied degradation of propamocarb hydrochloride and thiamethoxam was enhanced in the presence of B. amyloliquefaciens IN937a and B. pumilus SE34. Acibenzolar-S-methyl, propamocarb hydrochloride and thiamethoxam significantly increased the PGPR growth. However, the stimulatory effect was related to the level of pesticide spiked.

Determination of acibenzolar-S-methyl and its acidic metabolite in soils by HPLC-diode array detection.

A simple and accurate method for the analysis of acibenzolar-S-methyl (benzo[1,2,3]thiadiazole-7-carbothioic acid-S-methyl ester; CGA 245 704; ASM) and its major conversion product, benzo[1,2,3]thiadiazole-7-carboxylic acid (CGA 210 007; BTC), in soils is presented. ASM extraction from soil samples was performed using acetonitrile and BTC was extracted with a mixture of potassium phosphate buffer (0.5 M, pH 3) and acetonitrile (70:30 %, v/v). Both extracts were directly analyzed in a high-performance liquid chromatography-diode array detection (HPLC-DAD) system. Pesticide separation was achieved on a C18 (4.6 mm × 150 mm, 5 µm) analytical column with a isocratic elution of acetonitrile:water 40:60 % (v/v) with 0.6 mL L⁻¹ acetic acid at a flow rate of 1 mL min⁻¹. Linear regression coefficients (r (2)) of the external calibration curves were always above 0.9997. The limits of detection (LOD) and quantification (LOQ) of the method were 0.005 and 0.02 mg kg⁻¹ for ASM, and 0.01 and 0.05 mg kg⁻¹ for BTC, respectively. Recoveries were investigated at six fortification levels and were in the range of 90-120 % for ASM and 74-96 % for BTC with relative standard deviations (RSDs) below 11 % in all cases. The method was also validated by analyzing freshly spiked soil samples with 2.7% organic matter content at 0.5 mg kg⁻¹ level, with slightly lower recovery values only for ASM. Moreover, recoveries for intermediate aged residues of the analytes were similar to fresh residues. This method was also applied to determine ASM half-life (t(½) = 8.7 h) and the rate of the acidic metabolite formation.