Different working conditions shift the genetic damage levels of pesticide-exposed agriculture workers.

In the current study, we had two main purposes. Firstly, we aimed to compare genetic damages in the agricultural workers of two different types of environmental conditions including the greenhouse and open fields. Secondly, we aimed to compare genetic damages in the total agricultural workers as the exposed group (greenhouse and open field workers) (n = 114) and the non-exposed control group (n = 98) living in the same area in Canakkale, Turkey. For these purposes, we investigated the incidence of micronucleus (MN), nucleoplasmic bridges (NPBs), and nuclear buds (NBUDs) in peripheral blood lymphocytes. We observed that the frequencies of MN, NPB, and NBUD obtained for the greenhouse workers were statistically significantly higher than those obtained for the open field workers. When the results of the control group were compared with those of the total workers, there were statistically significant differences in terms of MN and NBUD frequencies. We found that age and MN were correlated at a significant level in both the agricultural workers and the control group. The MN frequency of the female workers was 1.5 times greater than that of the male workers, and it was a significant level in the agricultural workers.

Results of buccal micronucleus cytome assay in pesticide-exposed and non-exposed group.

Since many different pesticides have been used occupationally, there have been inconsistent results regarding DNA damages among greenhouse workers. Thus, the aim of the study is to evaluate DNA damages, cell death, and chromosomal instability by using the buccal micronucleus cytome (BMcyt) assay in greenhouse workers and to compare those with a non-exposed group. The BMcyt assay was applied to the exfoliated buccal cell samples collected from 66 pesticide-exposed and 50 non-exposed individuals. We evaluated the frequency of micronucleus (MN), nuclear bud (NBUD), binucleated (BN) cells, and karyolitic (KL), pyknotic (PY), and karyorrhectic (KH) cells. The results showed that the MN, BN, PY, and KH frequencies of the pesticide-exposed group were significantly higher than those of the controls (P Ë‚ 0.05, P Ë‚ 0.05, P Ë‚ 0.01, and P Ë‚ 0.05, respectively). We observed that the MN, BN, PY, and KH frequencies in the autumn were statistically different compared with those in the control group (P = 0.037 for MN, P = 0.001 for BN, P = 0.016 for PY, and P = 0.033 for KH). The same comparison was done in the spring for the control, and there was a statistically significant difference for MN (P = 0.046) and PY (P = 0.014). We can conclude that pesticide exposure in greenhouse workers was one of the factors that altered DNA damages, cell death, and chromosomal instability in oral mucosa cells.

Comet assay for assessment of DNA damage in greenhouse workers exposed to pesticides.

Purpose: The main goal of the present study was to determine DNA damage in pesticide-exposed greenhouse workers and pesticides non-exposed controls. Materials and methods: The DNA damage was measured by alkaline comet assay method (pH > 13) in 41 greenhouse workers and 45 non-exposed individuals as the control. Pesticide exposure was assessed by duration of working in the greenhouse and pesticide application in the greenhouse time. DNA damage was estimated by arbitrary unit and damage frequency. Results: Arbitrary unit and damage frequency were consistently significantly higher in greenhouse workers than those of the controls (p = 0.001). In terms of gender in greenhouse, DNA damage of female workers was significantly higher than those in male workers (p < 0.05). We found significant correlation between DNA damage and working hours spent. Multiple linear regression analysis showed that working hours in the greenhouse as an indication of pesticide exposure were significantly associated with the DNA damage, which can be attributed to the genotoxic potential of the pesticide mixture. Conclusions: The comet assay is sensitive to detect the damage exposed to chronic effect of pesticides in greenhouse workers. Significant DNA damage was obtained for the exposed group, which was associated with the pesticide exposure.

In vitro genotoxic and cytotoxic effects of doxepin and escitalopram on human peripheral lymphocytes.

Antidepressants are drugs used for the treatment of many psychiatric conditions including depression. There are findings suggesting that these drugs might have genotoxic, carcinogenic, and/or mutagenic effects. Therefore, the present in vitro study is intended to investigate potential genotoxic and cytotoxic effects of the antidepressants escitalopram (selective serotonin reuptake inhibitor) and doxepin (Tricyclic antidepressant) on human peripheral lymphocytes cytokinesis-block micronucleus (CBMN), sister chromatid exchange (SCE), and single cell gel electrophoresis (alkaline comet assay) were used for the purpose of the study. In the study, four different concentrations of both drugs (1, 2.5, 5, and 10 µg/mL) were administered to human peripheral lymphocytes for 24 h. The tested concentrations of both drugs were found to exhibit no cytotoxic and mitotic inhibitory effects. SCE increase caused by 5 and 10 µg/mL of escitalopram was found statistically significant, while no statistically significant increase was observed in DNA damage and micronucleus (MN) formation. Moreover, the increase caused by doxepin in MN formation was not found statistically significant. Besides, 10 µg/mL of doxepin was demonstrated to significantly increase arbitrary unit and SCE formation. These findings suggest that the investigated concentrations of escitalopram and doxepin were non-cytotoxic but potentially genotoxic at higher concentrations.

DNA damage and circulating cell free DNA in greenhouse workers exposed to pesticides.

Pesticides widely used in agriculture and other applications have been linked to cancer and other diseases through several potential mechanisms. The goals of this study were to assess DNA damage in lymphocytes by the cytokinesis-block micronucleus assay (CBMN), and to measure circulating cell free DNA (ccf-DNA) in the blood of pesticide-exposed greenhouse workers and matched controls living in the same area. CBMN was applied to peripheral blood lymphocyte samples taken at different times (spring and autumn) for each individual. We measured plasma ccf-DNA levels using a Qubit® fluorometer. The results indicated that the MNL, BNMN, and NBUDs frequencies of pesticide-exposed individuals were significantly higher than non-exposed individuals. Apart from MNL, BNMN and CBPI, a season-related effect was found for the NPB and NBUD frequencies. With MNL and BNMN as the dependent variables, multiple regression analysis showed that age and gender affected MN formation. The ccf-DNA level in the pesticide-exposed group was significantly higher than the control group. There was no seasonal variation regarding the free DNA amount. Ccf-DNA in males was found to be higher than females. The MNL, BNMN, NPB, and CBPI did not correlate with the ccf-DNA amount. It can be concluded that pesticide exposure can modulate DNA integrity via different mechanisms. Also, elevated levels of ccf-DNA could be recommended as a biomarker of pesticide exposure. Environ. Mol. Mutagen. 59:161-169, 2018. © 2017 Wiley Periodicals, Inc.

Genotoxicity of commercial fungicide Cabrio Plus on human cell.

Cabrio Plus, a commercial fungicide, is used in agriculture as the control agent for a broad spectrum of diseases including black dot, early blight, late blight and powdery mildew. This study aimed to evaluate the genotoxicity of commercial formulation of Cabrio Plus which has been inadequately evaluated. The genotoxic potential of Cabrio Plus in isolated human peripheral blood lymphocytes was measured by means of an alkaline version of the comet assay (pH > 13) and in whole blood by use of the in vitro micronucleus test. Cabrio Plus induced a statistically significant increase in DNA damage assessed with the in vitro micronucleus assay and the comet assay. Cabrio Plus also induced cytotoxicity in a dose-dependent manner with the in vitro micronucleus assay. It can be concluded that a commercially available pesticide formulation, Cabrio Plus, has the ability to cause DNA damage and cytotoxicity.

Micronuclei, nucleoplasmic bridges, and nuclear buds induced in human lymphocytes by the fungicide signum and its active ingredients (boscalid and pyraclostrobin).

The aim of this study was to investigate the genotoxic and cytotoxic potential of the Signum fungicide and its active ingredients (boscalid and pyraclostrobin) on human peripheral blood lymphocytes using the cytokinesis-block micronucleus (CBMN) assay. Micronuclei (MNi), nucleoplasmic bridges (NPBs), nuclear bud (NBUDs) formations, and the cytokinesis-block proliferation index (CBPI) were evaluated in treated lymphocytes in Go (cells were treated and then kept in culture without stimulation for 24 h) and proliferation phases (cells were treated after 44 h culture in medium containing phytohemagglutinin). MN formation in lymphocytes treated in G0 statistically increased at doses of 2, 6, and 25 µg/mL signum; 0.5 and 2 µg/mL boscalid; and 0.5, 1.5, and 2 µg/mL pyraclostrobin; while NPB formation increased at a dose of 0.25 µg/mL pyraclostrobin. All concentrations of each fungicide did not statistically increase NBUD formation, while the cytotoxicity increased the dependent on concentration in lymphocytes treated in G0 . Doses of 0.5, 1, 1.5, and 3 µg/mL signum; 0.5, 1, and 1.5 µg/mL boscalid; and 0.75 µg/mL pyraclostrobin statistically increased the MN formation in proliferating lymphocytes. NPB formation increased in proliferating lymphocytes at doses of 1, 1.5, 2, and 3 µg/mL signum and at a dose of 0.75 µg/mL pyraclostrobin. In addition, a dose of 0.75 µg/mL pyraclostrobin increased NBUD frequencies. Cytotoxicity increased with increasing concentrations of each fungicide. It is concluded that signum, boscalid, and pyraclostrobin may be genotoxic and cytotoxic in vitro human peripheral blood lymphocytes in consideration of each of the two protocols. © 2012 Wiley Periodicals, Inc. Environ Toxicol 29: 723-732, 2014.

Prevelance of common YMDD motif mutations in long term treated chronic HBV infections in a Turkish population.

In the current study we aimed to show the common YMDD motif mutations in viral polymerase gene in chronic hepatitis B patients during lamivudine and adefovir therapy. Forty-one serum samples obtained from chronic hepatitis B patients (24 male, 17 female; age range: 34-68 years) were included in the study. HBV-DNA was extracted from the peripheral blood of the patients using an extraction kit (Invisorb, Instant Spin DNA/ RNA Virus Mini Kit, Germany). A line probe assay and direct sequencing analyses (INNO-LIPA HBV DR v2; INNOGENETICS N.V, Ghent, Belgium) were applied to determine target mutations of the viral polymerase gene in positive HBV-DNA samples. A total of 41 mutations located in 21 different codons were detected in the current results. In 17 (41.5%) patients various point mutations were detected leading to lamivudin, adefovir and/ or combined drug resistance. Wild polymerase gene profiles were detected in 24 (58.5%) HBV positive patients of the current cohort. Eight of the 17 samples (19.5%) having rtM204V/I/A missense transition and/or transversion point mutations and resistance to lamivudin. Six of the the mutated samples (14.6%) having rtL180M missense transversion mutation and resistance to combined adefovir and lamivudin. Three of the mutated samples (7.5%) having rtG215H by the double base substituation and resistance to adefovir. Three of the mutated samples (7.5%) having codon rtL181W due to the missense transversion point mutations and showed resistance to combined adefovir and lamivudin. Unreported novel point mutations were detected in the different codons of polymerase gene region in the current HBV positive cohort fromTurkish population. The current results provide evidence that rtL180M and rtM204V/I/A mutations of HBV-DNA may be associated with a poor antiviral response and HBV chronicity during conventional therapy in Turkish patients.

Evaluation of background DNA damage in a Turkish population measured by means of the cytokinesis-block micronucleus cytome assay.

This study aimed to determine the background incidence of micronuclei (MN), nucleoplasmic bridges (NPBs), and nuclear buds (NBUDs) in the peripheral blood lymphocytes of a population in the province of Çanakkale, Turkey. Seventy-one individuals living under similar socio-economic conditions, not exposed to an X-ray examination during the last six months, receiving no medical treatment and not involved in agricultural activities were evaluated. In addition to genetic end-points, the cytokinesis-block proliferation index (CBPI) was determined. MN and NBUD frequencies gradually increased with age. MN frequencies of the older group were ∼7 fold higher than those of the younger group. Multiple linear regression analysis showed that apart from age, no significant effects of various confounding factors on MN and NBUD were observed. NPB frequencies were affected by gender. In the present study, no statistically significant effect was found of smoking on the genetic end-points evaluated, which agrees with results from other studies.

Evaluation of metal concentrations in mussel M. galloprovincialis in the Dardanelles Strait, Turkey in regard of safe human consumption.

Concentrations of the elements were evaluated for the first time in Mytilus galloprovincialis in the Dardanelles (Canakkale Strait-Turkey). The concentration of elements were measured in samples collected in 2007, 2008 and 2009, while the concentrations of Fe and Ni were evaluated in samples taken in 2009. The maximum concentrations of Cd, Cr, Cu, Pb, Zn, Fe, and Ni were found to be 1.59, 6.04, 12.01, 6.03, 319.6, 402.79, and 3.52 mg/kg, respectively. In terms of the nutritional aspect, taking into account the values recommended by world health authorities, the concentration of elements can generally be considered not to be at levels posing a health risk.

Frequencies of micronuclei (MNi), nucleoplasmic bridges (NPBs), and nuclear buds (NBUDs) in farmers exposed to pesticides in Çanakkale, Turkey.

This study aimed to determine the incidence of micronuclei (MNi), nucleoplasmic bridges (NPBs), and nuclear buds (NBUDs) in peripheral blood lymphocytes due to direct exposure to pesticides among 46 farmers in Çanakkale, Turkey. 48 non-exposed individuals living in the same socioeconomic conditions were chosen as control. In addition, a cytokinesis-block proliferation index (CBPI) was calculated. MNi and NBUDs frequencies were significantly higher among the farmers (p < 0.05). Although the NPB frequency of the farmers was higher than the controls, there was no statistical difference. Multiple linear regression analysis showed that apart from gender, no significant effects of various confounding factors were observed. Regarding CBPI, data obtained for the controls were higher than that of the farmers; however, there was no statistically significant difference.

Heavy metal deposition in moss samples from east and south Marmara region, Turkey.

A survey of atmospheric heavy metal deposition in the east and south Marmara region, Turkey was carried out in September 2004. For this purpose, moss samples (Hypnum cupressiforme) were collected in a systematic network of 125 sites. Concentrations of the elements (Al, B, Ba, Ca, Cd, Fe, Co, Cr, Cu, K, Li, Mn, Na, Ni, Pb, Sr, Mg, Ti, and Zn) in the moss were used as an indication of the level of air pollution in the region. Significant differences in heavy metal concentrations, especially for Pb, Cd, Cu, and Zn, were recorded in the moss samples collected around industrialized and heavily populated cities (Istanbul, Bursa, Bandirma, Kocaeli, Biga-Çan) and in an abandoned lead-mining area (Balikesir-Balya). A map of the spatial distribution of each element in the region was plotted, and enrichment factors were calculated. VARIMAX principal component analysis was applied to the data obtained, and five different components were obtained. The results showed that Pb, Cu, Cd, and Zn derived from anthropogenic and industrial sources while other elements came mostly from natural sources.

The heavy metal content of wild edible mushroom samples collected in canakkale province, Turkey.

In this study, concentrations of heavy metals (Cd, Cr, Cu, Pb, and Zn) were determined in three edible mushroom species (Lactarius deliciosus, Russula delica, and Rhizopogon roseolus) collected in five sampling sites in Canakkale province, Turkey. Mean values of Cd, Cr, Cu, Pb, and Zn were 0.72, 0.26, 28.34, 1.53, and 64.62 mg/kg, respectively. The highest concentrations of Cd, Cu, Pb, and Zn were determined in species R. delica, while Cr was observed in L. deliciosus. In terms of the nutritional aspect, taking into account the concentration of Provisional Tolerable Weekly Intake recommended by FAO/WHO, the maximum concentration of Cd is a restrictive factor for consumption of the collected mushroom species. The concentrations of the other elements have no health risks when consumed at optimal levels.

[The serological and entomological survey of zoonotic visceral leishmaniasis in Ayvacik Region of Canakkale Province, Turkey]. / Canakkale Ili Ayvacik Bölgesinde Zoonotik Visseral Leishmaniasisin Serolojik ve Entomolojik Olarak Arastirilmasi.

The field studies were carried out in Ilyasfaki village belonging to Ayvacik town, Kalabakli village and center of Kepez town belonging to Canakkale province to determine the epidemiological situation of visceral leishmaniasis between June and August 2007. A total of 27 blood samples were taken from the dogs which are known as reservoir of Leishmania infantum causing visceral leishmaniasis in human and dogs. The dogs were also examined physically for clinical symptoms of the disease. A total of 789 sand fly specimens were collected using CDC miniature light traps and examined. Six Phlebotomus and one Sergentomyia species were identified as: Phlebotomus negletus, P. tobbi, P. simici, P. papatisi, P. perfiliewi, P. halepensis and S. theodori. Among these species P. neglectus, was found to be a dominant species in Ilyasfaki village by the rate of 94.4%, while P. tobbi was also found to be a dominant species in city center (including Kepez) and Kalabaki village by the rate of 50% and 48.1%, respectively. No seropositive dogs were found by IFA test. Seropositivity below cut-off level was only observed in two dogs from Kepez, in dilutions of 1/16 and 1/64. The risk level for human and canine visceral is found to be high and it requires more field studies on canine leishmaniasis in and around the area.

Cross-border response of mosses to heavy metal atmospheric deposition in Southeastern Bulgaria and European Turkey.

First cross-border atmospheric pollution of 11 heavy metals and toxic elements assessed by Hypnum cupressiforme was reported for a part of Southeastern Europe (Southeastern Bulgaria and European Turkey). Moss monitoring technique followed the main requirements of European Moss Survey. Moss samples were collected in April 2006 both in Bulgaria and Turkey. Concentration of Al, As, Cd, Cr, Cu, Fe, Ni, Pb, Sb, V, and Zn were determined by ICP-AES. Interlaboratory parallel calibration (exchanged four moss samples from each country), standard reference moss materials (M2 and M3) results ensured the study. ANOVA showed no differences between measured results in both laboratories at the 99% confidence level. Principle Component Analyze proved two factors: F1 group of Al, As, Cd, Cr, Fe, Ni, and V and F2 of Cu, Pb, and Zn as main atmospheric pollutants. Results obtained showed approximately Cu and Pb high concentrations around Istanbul and Burgas and Zn pollution in Istanbul district. Arsenic cross-border atmospheric pollution in the study area of Southeastern Europe was found.

Comparison of epigeic moss (Hypnum cupressiforme) and lichen (Cladonia rangiformis) as biomonitor species of atmospheric metal deposition.

In the present work epigeic moss (Hypnum cupressiforme Hedw.) and epigeic lichen (Cladonia rangiformis Hoffm.) samples were collected simultaneously in the Thrace region, Turkey according to a regular sampling grid. Whereas the moss was found at all 68 sampling sites, the lichen could be collected only at 25 of the sites, presumably because lichens are more sensitive than mosses with respect to air pollution and climatic variations. All elements showed higher accumulation in the moss than in the lichen whereas element inter-correlations were generally higher in the lichen. All considered the moss was judged to be a better choice than the lichen for biomonitoring of atmospheric deposition of metals in this case, and it is argued that mosses may be generally more suited than lichens for this purpose.

Radioactivity concentrations and dose assessment in surface soil samples from east and south of Marmara region, Turkey.

The activity concentrations of 137Cs, 40K, 232Th, 238U and 226Ra were measured in surface soil samples from East and South of Marmara region, Turkey. The physico-chemical parameters (organic matter, CaCO3 contents and pH-value) of the soil samples were determined in the samples collected from 100 sampling stations. The average activity concentrations of 137Cs, 40K, 232Th, 238U and 226Ra were found to be 27.46+/-21.84, 442.51+/-189.85, 26.63+/-15.90, 21.77+/-12.08 and 22.45+/-13.31 Bq kg(-1), respectively. The mean value of total annual external gamma radiation dose equivalent for the natural radionuclides was calculated to be 54.86 microSv. The current data were compared with those found in the other locations of Turkey and different countries.

Determination of atmospheric heavy metal pollution in Canakkale and Balikesir provinces using lichen (Cladonia rangiformis) as a bioindicator.

A regional study was conducted to assess the current level of atmospheric heavy metal pollution (Pb, Zn, Cr, Cu, Cd) in the Canakkale and Balikesir provinces of Turkey, and also to establish a baseline for future studies of atmospheric heavy metal pollution. The lichen Cladonia rangiformis was used for determining the distribution of heavy metals in the atmosphere. The highest concentration of all these elements was observed in the vicinity of Balikesir province where there is an abandoned lead-zinc main. On the other hand, apart from the mining area, the concentration of heavy metals was similar to the data reported for unpolluted areas. Maximum values of Pb, Zn, Cr, Cu, and Cd were 33.8, 47.6, 13.0, 5.29, and 0.69 mg/kg in dry weight, respectively. Spatial distribution of these elements, apart from Cr, was similar. Correlation coefficients between Zn-Pb, Cr-Zn, Cu-Pb, Cu-Zn, Cd-Zn, and Cd-Cr were high and positive, and indicated that they come from the same sources.

Toxic metals in the Austrian pine (pinus nigra) bark in the Thrace region, Turkey.

Austrian pine (Pinus nigra) bark was collected in thirty-seven localities in the Thrace region, Turkey, for the determination of airborne toxic metal distribution. Five toxic metals, As, Cd, Cu, Pb, and Zn, were monitored using atomic absorption spectrometry. Distribution maps of toxic elements in the region were plotted according to Geographic Information System technology. In general, the concentrations of all investigated elements were not high in the region apart from the vicinity of big cities, Tekirdag, Istanbul and, close to Bulgarian border. However, the highest concentrations were not at pollution level. They have a similar distribution pattern throughout the region.