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Influence of organic solvents upon the toxicity of deltamethrin
Abstracts / Toxicology Letters 180S (2008) S32–S246
Helicoverpa armigera, cotton bollworm, is a polyphagous insect. It is one of the most important preharvest pests for economically important crops including cotton, legumes, and vegetables. In this study, midguts of H. armigera obtained from Adana, Antalya, C¸anakkale and Mardin field populations were used. GST activities were determined using 1-chloro-2,4-dinitrobenzene as a substrate. Adana and Antalya field populations showed statistically significant (p < 0.05), 2.4 and 2.9 fold increase, respectively, compared to susceptible population. General esterase activity measurements were done with Adana, Antalya and susceptible populations by using p-nitro phenyl acetate as a substrate. Adana population showed statistically significant increase (p < 0.05); nevertheless, a small increment (1.29 fold) was found as compared to susceptible population. Although, Antalya population did not show statistically significant increase (p > 0.05) compared to susceptible population, it exhibited tendency to increase. The role of cytochrome P-450 monooxygenases (CYP6B6, CYP6B7, and CYP9A12) in pyrethroid resistance development analyzed using Reverse Transcriptase-PCR method. In these experiments, C¸anakkale and Mardin field populations of H. armigera were used. Some individuals in both field populations exhibited slight increases in gene expressions of CYP6B7 and CYP9A12 genes compared to susceptible individuals. CYP6B6 gene expression was not detected in susceptible strain; nonetheless, expression of the same gene was observed in some of the individuals from Mardin and C¸anakkale field populations. Increased GST activity in conjunction with activation of CYP6B6 appears to play a role in the resistance of Turkish H. armigera in monitored fields. Acknowledgements: This work was financially supported by project no: BAP-DPT2002K120510. doi:10.1016/j.toxlet.2008.06.270 S17 Identification and determination of herbicide residues in wheat by HPLC Sana Hajy Shah 1,2,∗ , Zahra Nazari Khorasgani 1,2 , Nooshin Asad Masjedi 1,2 1
College of Agricultural, Ahvaz Shahid Chamran University, Ahvaz/Khouzestan, Islamic Republic of Iran, 2 Department of Toxicology & Pharmacology, Pharmacy School, Ahvaz Jondishapour University of Medical Sciences, Ahvaz/Khouzestan, Islamic Republic of Iran Wheat is the key ingredient of bread, which is a staple food in many countries, Iran ranked ninth in terms of grains production. Atrazine and amytrine are not used directly on wheat but they are used for control of volunteer wheat in certain production systems and have been classified as possible human carcinogens. Since, there was not sufficient available data about pesticide residues in distributed wheat in flour factories of Ahvaz city at Khozestan province in Iran, this study was done. For this purpose 32 wheat samples of 4 flour factories (Mahziyar, Ahvaz, Khouzestan, and Arde Jonob) were taken. 20 g of each collected samples was macerated in 50 ml water. After adjusting pH to <2 and adding 5 g sodium chloride was mixed and filtrated. The filtrated was percolated through C18 cartridge which was conditioned with 10 ml methanol and 10 ml of water, respectively under vacuum. Then the column washed with 5 ml of water, and the elute discarded, the sorbent bed was dried and finally the pesticides in all of the samples were eluted with 1 ml of methanol. The solvent was evaporated under nitrogen gas. The residue was dissolved in 0.5 ml of mobile phase of methanol–water (60:40) at pH 4.6 and was analyzed by HPLC using
S169
a C18 column with a UV detector at 230 nm. The mean concentrations of atrazine and amytrine in investigated wheat samples were 0.1 and 0.299 g/kg, respectively. The samples did not exceed the maximum permissible levels for atrazine and amytrine and were safe for human consumption. doi:10.1016/j.toxlet.2008.06.271 S18 Assessment of hazard for human of herbicides, belonging to the triketones class, during their application on radioactively polluted territories Nataliia Kovalchuk ∗ , Vasyl Bardov, Sergiy Sasinovych, Igor Pelo, Tetiana Girenko
Omelchuk, Larysa
Institute of Hygiene and Ecology of National O.O. Bohomolets Medical University, Kyiv, Ukraine Stable trend of increasing of the thyroid pathology level has appeared during last years around the world and in Ukraine. This problem is especially important for Ukrainian population, who have been affected by the radioactive iodine (I131) after Chornobyl accident. Due to this, negative influence of any factors may affect the thyroid gland. Wide introduction of new triketone herbicides into the agriculture leads to necessity for the advanced assessment of their possible application on radioactively polluted territories. Triketones inhibit the HPPD (para-hydroxyphenylpyruvate dioxigenase) activity, which increases the tyrosine level in blood serum. The tyrosine degradation depends on the activity of two enzymes-HPPD and TAT (tyrosineaminotranspherase) in the blood serum. If activity of the HPPD is inhibited, but the TAT activity is sufficient, the tyrosine is not accumulated and no toxic effects appear. The TAT activity varies significantly depending on the animal species and sex. In the same experimental conditions organism of the mouse does not suffer such toxic effects as the organism of the rat. The value of the TAT activity in the human blood serum is comparable to that for mice. During the assessment of the triketone hazard, extrapolation of results, revealed in different experiments on mice, to human is the most adequate. Based on the results of experiments and field observations, received in real conditions, conclusions may be drawn. The application of triketone herbicides will not present additional hazard for workers on radioactively polluted territories, if all regulations and standards are adhered to. doi:10.1016/j.toxlet.2008.06.272 S19 Influence of organic solvents upon the toxicity of deltamethrin Mykola Prodanchuk ∗ , Igor Leposhkin, Nadiya Mykola Kokalko, Svitlana Yushchuk
Nedopytanska,
L.I. Medved Institute of Ecohygeine and Toxicology (ECOHYNTOX), Kiev, Ukraine In many cases hazard of pesticide product closely connect with used component. This problem taking into account presence in the market a bulk of generic formulation is very important. We studied the influence of different solvent on the toxicity of insecticide deltamethrin. Investigation was carried out on male rat’s administrated orally seven formulations of deltamethrin differing by vehicle. QSAR have been evaluated for predicting an acute (LD50 ) toxicity of deltamethrin formulation on the basis of physicochemical parameters of solvents.
S170
Abstracts / Toxicology Letters 180S (2008) S32–S246
We have received new data about relationship between toxicity of deltamethrin formulation and used organic solvents. As a result of our experiment it have been shown that the indices of toxicities (LD50 ) vary in a wide range and depends from solvents. The acute toxicities of formulation depend from the degree of solubility of active ingredient and correlates with molecular mass, lipophility and empirical parameters (solubility and polarity). Obtained relationship may be useful for more accurate selection of organic solvent for pesticide formulation from the point of view of their hazard. doi:10.1016/j.toxlet.2008.06.273 S20 Pattern of human acute intoxication in Egypt with special concern to pesticide poisoning and farmers’ attitude regarding use of these substances Sameeh Mansour 1,2,∗ , Hany Gamalludin 1,2 1
Pesticide Chemistry Department, National Research Centre, Dokki, Cairo, Egypt, 2 Poison Control Center, Ain Shams University (PCCA), Cairo, Egypt
Poisoning, either unintentional or intentional, is not confined to developing countries, it frequently occurs in developed countries as well. Over a 17 years period (1966–1982), a total number of 20,300 poisoned cases were recorded in Egypt; giving rise to an average of 1194 pesticide-poisoned persons/year. During the recent years (2003–2006), the number of poisoned cases admitted to the (PCCA) ranged between 21,805 and 25,555 cases/year. The great majority of poisonings (94.0%) occurred via the oral route. Non-drug substances accounted for 54.0% of poisonings, while drugs represented 37.0%. Poisoning by non-drugs included chemicals (60.0%), food poisoning (28.0%), animal poisoning (6.0%), and others. The main chemicals causing poisoning were: insecticides, corrosives, petroleum distillates and other chemicals, representing 51.0%, 22.0%, 13.0% and 14.0%, respectively. The number of pesticide-poisoned cases in 2006 (3564 persons) reached ca 3 times those occurred annually during the past decades. Of this number, OP insecticides accounted for 75.0%, while carbamates (5.0%) and zinc phosphide (20.0%) accounted for fewer poisoning incidents. In an attempt to investigate the attitude and behavior of the Egyptian farmers regarding pesticide use, the results revealed that safety measures are generally poorly applied and workers lack proper knowledge or training in safe handling of pesticides. It was possible to estimate the magnitude of occupational exposure among the studied sample of farmers. Accordingly, they were classified into five groups; e.g., excessive exposure (15.3%), extreme exposure (30.5%), high exposure (33.0%), moderate exposure (18.2%) and slight exposure (3.0%). Statistical analyses for the up-to-date data of the (PCCA) will be presented in details. doi:10.1016/j.toxlet.2008.06.274 S21 Application of different biotests to assess biological activity of organochlorine pesticides and their radiation metabolites Tatiana Melnikova ∗ , Lyudmila Polyakova, Gennady Kozmin State University of Nuclear Power Engineering, Obninsk/Kaluga, Russian Federation The problem of assessing the resistance of organochlorine pesticides (OCP) to gamma-irradiation is assuming a practical importance in view of the use of radiation technologies in the food
industry. As is evident from a literature overview, a certain OCP hazard to biological objects results from their low-level chronic effects. According to the previous studies, low OCP doses cause serious damage to a living organism, comparable with that from high doses. After radiolysis of OCPs, a variety of different structures have been found in model solutions. Though each is in trace concentrations, these constitute a major part of pollution (up to 90% of the initial OCP). Toxicologic effects of this pollution will be most likely dependent on a synergistic, or antagonistic, or combined effect, rather than that from dose variation of an individual component. Proliferative capacity of barley cells grown in Petri dishes was found to be informative for bioactivity assessment of OCP and their metabolites. The above substances were found to inhibit barley growth, the effect being maximally evident in overall root length. Barley growth stimulation (by seedling mass) was observed in seeds grown in soil. Significant difference of biological activity between irradiated and non-irradiated OCP solutions has been reported in other biological tests (by indicators such as chrorophyll destruction and spontaneous motor activity of spirostoma). It is assumed that biological effect from OCP irradiation can result from a synergistic action of different components of a disperse system (matrix pollutant), namely: residual pesticide, its metabolites and substances resulting from radiolysis of the pesticide, and the environment. The results derived are useful for developing recommendations for food quality organizations, according to which radiation treated food and agricultural products need to be analyzed not only for the presence of known OCP but also of their radiation metabolites. Taking into account simplicity and informative value of a biotest with the use of spirostoma, it is recommended that OCP studies be continued in search for the screening methods of their radiation metabolites. doi:10.1016/j.toxlet.2008.06.275 S22 Assessment of oxidative DNA damage by glyphosate applying hOGG1 modified comet and micronucleus assay Marin Mladinic ∗ , Davor Zeljezic Institute for Medical Research and Occupational Health, Zagreb, Croatia Genetically modified crops resistant to glyphosate are being used more widely each year. The modification enables them to tolerate higher concentrations of the active ingredient. Thus, higher amounts of glyphosate are introduced into the environment. In our study we tested concentrations that are likely to be encountered in residential and occupational exposure (0.5, 2.91, 3.5 and 580 g/ml). Evaluation was performed on human peripheral blood lymphocytes form three healthy, young volunteers. Technical glyphosate (98%) was tested with and without metabolic activation system (S9). Lymphocytes were treated for 4 h at 37 ◦ C. Alkaline comet assay was modified by applying hOGG1 enzyme to detect 8-hydroxy-2 -deoxyguanosine as a result of oxidative DNA damage. Simultaneously, treated lymphocytes were used to set up cultures for micronucleus assay (MN). Micronuclei slides were hybridized with All Human Centromere Satellite Probes. The hOGG1 comet assay without S9 showed significant increase of the tail intensity only at 3.5 g/ml, but not at highest concentration tested (580 g/ml). The presence of S9 significantly elevated the tail length only at the 580 g/ml. Altogether, the DNA damage detected was not dose dependent. Significant number of MN was observed only at the highest concentration when applying S9. Also, only for that treatment proportion of MN with centromere signal was
Helicoverpa armigera, cotton bollworm, is a polyphagous insect. It is one of the most important preharvest pests for economically important crops including cotton, legumes, and vegetables. In this study, midguts of H. armigera obtained from Adana, Antalya, C¸anakkale and Mardin field populations were used. GST activities were determined using 1-chloro-2,4-dinitrobenzene as a substrate. Adana and Antalya field populations showed statistically significant (p < 0.05), 2.4 and 2.9 fold increase, respectively, compared to susceptible population. General esterase activity measurements were done with Adana, Antalya and susceptible populations by using p-nitro phenyl acetate as a substrate. Adana population showed statistically significant increase (p < 0.05); nevertheless, a small increment (1.29 fold) was found as compared to susceptible population. Although, Antalya population did not show statistically significant increase (p > 0.05) compared to susceptible population, it exhibited tendency to increase. The role of cytochrome P-450 monooxygenases (CYP6B6, CYP6B7, and CYP9A12) in pyrethroid resistance development analyzed using Reverse Transcriptase-PCR method. In these experiments, C¸anakkale and Mardin field populations of H. armigera were used. Some individuals in both field populations exhibited slight increases in gene expressions of CYP6B7 and CYP9A12 genes compared to susceptible individuals. CYP6B6 gene expression was not detected in susceptible strain; nonetheless, expression of the same gene was observed in some of the individuals from Mardin and C¸anakkale field populations. Increased GST activity in conjunction with activation of CYP6B6 appears to play a role in the resistance of Turkish H. armigera in monitored fields. Acknowledgements: This work was financially supported by project no: BAP-DPT2002K120510. doi:10.1016/j.toxlet.2008.06.270 S17 Identification and determination of herbicide residues in wheat by HPLC Sana Hajy Shah 1,2,∗ , Zahra Nazari Khorasgani 1,2 , Nooshin Asad Masjedi 1,2 1
College of Agricultural, Ahvaz Shahid Chamran University, Ahvaz/Khouzestan, Islamic Republic of Iran, 2 Department of Toxicology & Pharmacology, Pharmacy School, Ahvaz Jondishapour University of Medical Sciences, Ahvaz/Khouzestan, Islamic Republic of Iran Wheat is the key ingredient of bread, which is a staple food in many countries, Iran ranked ninth in terms of grains production. Atrazine and amytrine are not used directly on wheat but they are used for control of volunteer wheat in certain production systems and have been classified as possible human carcinogens. Since, there was not sufficient available data about pesticide residues in distributed wheat in flour factories of Ahvaz city at Khozestan province in Iran, this study was done. For this purpose 32 wheat samples of 4 flour factories (Mahziyar, Ahvaz, Khouzestan, and Arde Jonob) were taken. 20 g of each collected samples was macerated in 50 ml water. After adjusting pH to <2 and adding 5 g sodium chloride was mixed and filtrated. The filtrated was percolated through C18 cartridge which was conditioned with 10 ml methanol and 10 ml of water, respectively under vacuum. Then the column washed with 5 ml of water, and the elute discarded, the sorbent bed was dried and finally the pesticides in all of the samples were eluted with 1 ml of methanol. The solvent was evaporated under nitrogen gas. The residue was dissolved in 0.5 ml of mobile phase of methanol–water (60:40) at pH 4.6 and was analyzed by HPLC using
S169
a C18 column with a UV detector at 230 nm. The mean concentrations of atrazine and amytrine in investigated wheat samples were 0.1 and 0.299 g/kg, respectively. The samples did not exceed the maximum permissible levels for atrazine and amytrine and were safe for human consumption. doi:10.1016/j.toxlet.2008.06.271 S18 Assessment of hazard for human of herbicides, belonging to the triketones class, during their application on radioactively polluted territories Nataliia Kovalchuk ∗ , Vasyl Bardov, Sergiy Sasinovych, Igor Pelo, Tetiana Girenko
Omelchuk, Larysa
Institute of Hygiene and Ecology of National O.O. Bohomolets Medical University, Kyiv, Ukraine Stable trend of increasing of the thyroid pathology level has appeared during last years around the world and in Ukraine. This problem is especially important for Ukrainian population, who have been affected by the radioactive iodine (I131) after Chornobyl accident. Due to this, negative influence of any factors may affect the thyroid gland. Wide introduction of new triketone herbicides into the agriculture leads to necessity for the advanced assessment of their possible application on radioactively polluted territories. Triketones inhibit the HPPD (para-hydroxyphenylpyruvate dioxigenase) activity, which increases the tyrosine level in blood serum. The tyrosine degradation depends on the activity of two enzymes-HPPD and TAT (tyrosineaminotranspherase) in the blood serum. If activity of the HPPD is inhibited, but the TAT activity is sufficient, the tyrosine is not accumulated and no toxic effects appear. The TAT activity varies significantly depending on the animal species and sex. In the same experimental conditions organism of the mouse does not suffer such toxic effects as the organism of the rat. The value of the TAT activity in the human blood serum is comparable to that for mice. During the assessment of the triketone hazard, extrapolation of results, revealed in different experiments on mice, to human is the most adequate. Based on the results of experiments and field observations, received in real conditions, conclusions may be drawn. The application of triketone herbicides will not present additional hazard for workers on radioactively polluted territories, if all regulations and standards are adhered to. doi:10.1016/j.toxlet.2008.06.272 S19 Influence of organic solvents upon the toxicity of deltamethrin Mykola Prodanchuk ∗ , Igor Leposhkin, Nadiya Mykola Kokalko, Svitlana Yushchuk
Nedopytanska,
L.I. Medved Institute of Ecohygeine and Toxicology (ECOHYNTOX), Kiev, Ukraine In many cases hazard of pesticide product closely connect with used component. This problem taking into account presence in the market a bulk of generic formulation is very important. We studied the influence of different solvent on the toxicity of insecticide deltamethrin. Investigation was carried out on male rat’s administrated orally seven formulations of deltamethrin differing by vehicle. QSAR have been evaluated for predicting an acute (LD50 ) toxicity of deltamethrin formulation on the basis of physicochemical parameters of solvents.
S170
Abstracts / Toxicology Letters 180S (2008) S32–S246
We have received new data about relationship between toxicity of deltamethrin formulation and used organic solvents. As a result of our experiment it have been shown that the indices of toxicities (LD50 ) vary in a wide range and depends from solvents. The acute toxicities of formulation depend from the degree of solubility of active ingredient and correlates with molecular mass, lipophility and empirical parameters (solubility and polarity). Obtained relationship may be useful for more accurate selection of organic solvent for pesticide formulation from the point of view of their hazard. doi:10.1016/j.toxlet.2008.06.273 S20 Pattern of human acute intoxication in Egypt with special concern to pesticide poisoning and farmers’ attitude regarding use of these substances Sameeh Mansour 1,2,∗ , Hany Gamalludin 1,2 1
Pesticide Chemistry Department, National Research Centre, Dokki, Cairo, Egypt, 2 Poison Control Center, Ain Shams University (PCCA), Cairo, Egypt
Poisoning, either unintentional or intentional, is not confined to developing countries, it frequently occurs in developed countries as well. Over a 17 years period (1966–1982), a total number of 20,300 poisoned cases were recorded in Egypt; giving rise to an average of 1194 pesticide-poisoned persons/year. During the recent years (2003–2006), the number of poisoned cases admitted to the (PCCA) ranged between 21,805 and 25,555 cases/year. The great majority of poisonings (94.0%) occurred via the oral route. Non-drug substances accounted for 54.0% of poisonings, while drugs represented 37.0%. Poisoning by non-drugs included chemicals (60.0%), food poisoning (28.0%), animal poisoning (6.0%), and others. The main chemicals causing poisoning were: insecticides, corrosives, petroleum distillates and other chemicals, representing 51.0%, 22.0%, 13.0% and 14.0%, respectively. The number of pesticide-poisoned cases in 2006 (3564 persons) reached ca 3 times those occurred annually during the past decades. Of this number, OP insecticides accounted for 75.0%, while carbamates (5.0%) and zinc phosphide (20.0%) accounted for fewer poisoning incidents. In an attempt to investigate the attitude and behavior of the Egyptian farmers regarding pesticide use, the results revealed that safety measures are generally poorly applied and workers lack proper knowledge or training in safe handling of pesticides. It was possible to estimate the magnitude of occupational exposure among the studied sample of farmers. Accordingly, they were classified into five groups; e.g., excessive exposure (15.3%), extreme exposure (30.5%), high exposure (33.0%), moderate exposure (18.2%) and slight exposure (3.0%). Statistical analyses for the up-to-date data of the (PCCA) will be presented in details. doi:10.1016/j.toxlet.2008.06.274 S21 Application of different biotests to assess biological activity of organochlorine pesticides and their radiation metabolites Tatiana Melnikova ∗ , Lyudmila Polyakova, Gennady Kozmin State University of Nuclear Power Engineering, Obninsk/Kaluga, Russian Federation The problem of assessing the resistance of organochlorine pesticides (OCP) to gamma-irradiation is assuming a practical importance in view of the use of radiation technologies in the food
industry. As is evident from a literature overview, a certain OCP hazard to biological objects results from their low-level chronic effects. According to the previous studies, low OCP doses cause serious damage to a living organism, comparable with that from high doses. After radiolysis of OCPs, a variety of different structures have been found in model solutions. Though each is in trace concentrations, these constitute a major part of pollution (up to 90% of the initial OCP). Toxicologic effects of this pollution will be most likely dependent on a synergistic, or antagonistic, or combined effect, rather than that from dose variation of an individual component. Proliferative capacity of barley cells grown in Petri dishes was found to be informative for bioactivity assessment of OCP and their metabolites. The above substances were found to inhibit barley growth, the effect being maximally evident in overall root length. Barley growth stimulation (by seedling mass) was observed in seeds grown in soil. Significant difference of biological activity between irradiated and non-irradiated OCP solutions has been reported in other biological tests (by indicators such as chrorophyll destruction and spontaneous motor activity of spirostoma). It is assumed that biological effect from OCP irradiation can result from a synergistic action of different components of a disperse system (matrix pollutant), namely: residual pesticide, its metabolites and substances resulting from radiolysis of the pesticide, and the environment. The results derived are useful for developing recommendations for food quality organizations, according to which radiation treated food and agricultural products need to be analyzed not only for the presence of known OCP but also of their radiation metabolites. Taking into account simplicity and informative value of a biotest with the use of spirostoma, it is recommended that OCP studies be continued in search for the screening methods of their radiation metabolites. doi:10.1016/j.toxlet.2008.06.275 S22 Assessment of oxidative DNA damage by glyphosate applying hOGG1 modified comet and micronucleus assay Marin Mladinic ∗ , Davor Zeljezic Institute for Medical Research and Occupational Health, Zagreb, Croatia Genetically modified crops resistant to glyphosate are being used more widely each year. The modification enables them to tolerate higher concentrations of the active ingredient. Thus, higher amounts of glyphosate are introduced into the environment. In our study we tested concentrations that are likely to be encountered in residential and occupational exposure (0.5, 2.91, 3.5 and 580 g/ml). Evaluation was performed on human peripheral blood lymphocytes form three healthy, young volunteers. Technical glyphosate (98%) was tested with and without metabolic activation system (S9). Lymphocytes were treated for 4 h at 37 ◦ C. Alkaline comet assay was modified by applying hOGG1 enzyme to detect 8-hydroxy-2 -deoxyguanosine as a result of oxidative DNA damage. Simultaneously, treated lymphocytes were used to set up cultures for micronucleus assay (MN). Micronuclei slides were hybridized with All Human Centromere Satellite Probes. The hOGG1 comet assay without S9 showed significant increase of the tail intensity only at 3.5 g/ml, but not at highest concentration tested (580 g/ml). The presence of S9 significantly elevated the tail length only at the 580 g/ml. Altogether, the DNA damage detected was not dose dependent. Significant number of MN was observed only at the highest concentration when applying S9. Also, only for that treatment proportion of MN with centromere signal was