Drinking water quality: An in vitro approach for the assessment of cytotoxic and genotoxic load in water sampled along distribution system

An in vitro approach was performed to assess the quality of drinking water collected at two treatment/distribution networks located near the source (Plant #1) and the mouth of River Po (Plant #2). The water was sampled at different points of each distribution network, before (raw water) and after the chlorine dioxide disinfection, and in two points of the pipeline system to evaluate the influence of the distribution system on the amount and quality of the disinfection by-product. Cytotoxicity and genotoxicity of water extracts were evaluated in human peripheral lymphocytes and Hep-G2 cells by the use of the micronucleus (MN) test and Comet assay. Raw water samples of both plants induced cytotoxic effects, but not the increases of MN frequency in Hep-G2 cells and in human lymphocytes. Increases of DNA damage in human leukocytes was detected by Comet assay for raw water of Plant #2 at concentration ≥ 0.25 Leq/mL. The disinfection process generally has reduced the toxicity of water samples, even if potential direct DNA-damaging compounds have been detectable in drinking water samples. The proposal approach, if currently used together with chemical analysis, can contribute to improve the monitoring drinking water.     

Genotoxic effects of occupational exposure to lead and influence of polymorphisms in genes involved in lead toxicokinetics and in DNA repair

Lead is still widely used in many industrial processes and is very persistent in the environment. Although toxic effects caused by occupational exposure to lead have been extensively studied, there are still conflicting results regarding its genotoxicity. In a previous pilot study we observed some genotoxic effects in a population of lead exposed workers. Thus, we extended our study analysing a larger population, increasing the number of genotoxicity endpoints, and including a set of 20 genetic polymorphisms related to lead toxicokinetics and DNA repair as susceptibility biomarkers. Our population comprised 148 workers from two Portuguese factories and 107 controls. The parameters analysed were: blood lead levels (BLL) and δ-aminolevulinic acid dehydratase (ALAD) activity as exposure biomarkers, and T-cell receptor (TCR) mutation assay, micronucleus (MN) test, comet assay and OGG1-modified comet assay as genotoxicity biomarkers. Lead exposed workers showed markedly higher BLL and lower ALAD activity than the controls, and significant increases of TCR mutation frequency (TCR-Mf), MN rate and DNA damage. Oxidative damage did not experience any significant alteration in the exposed population. Besides, significant influence was observed for VDR rs1544410 polymorphism on BLL; APE1 rs1130409 and LIG4 rs1805388 polymorphisms on TCR-Mf; MUTYH rs3219489, XRCC4 rs28360135 and LIG4 rs1805388 polymorphisms on comet assay parameter; and OGG1 rs1052133 and XRCC4 rs28360135 polymorphisms on oxidative damage. Our results showed genotoxic effects related to occupational lead exposure to levels under the Portuguese regulation limit of 70μg/dl. Moreover, a significant influence of polymorphisms in genes involved in DNA repair on genotoxicity biomarkers was observed.     

Genotoxic biomonitoring of agricultural workers exposed to pesticides in the north of Sinaloa State, Mexico

Genotoxic damage was evaluated in 70 agricultural workers, 25 women and 45 men, exposed to pesticides in Las Grullas, Ahome, Sinaloa, Mexico, with an average of 7 years of exposure. The effect was detected through the sister chromatid exchanges (SCE) in lymphocytes of peripheral blood and micronuclei (MN) and other nuclear anomalies (NA) in buccal exfoliated cells. Also, the influence on cellular proliferation kinetics (CPK) was studied by means of the replication index (RI) and the cytotoxic effect was examined with the mitotic index (MI). The non-exposed group consisted of 70 other persons, 21 women and 47 men from the city of Los Mochis, Sinaloa, Mexico. Significant differences between the exposed and the non-exposed groups were observed in SCE, CPK, MI, MN and NA. Analysis of variance revealed that age, gender, smoking and alcohol consumption did not have a significant effect on genetic damage. However, there was a correlation between exposure time to pesticides and SCE frequency. These results could have been due to the exposure of workers to pesticides containing different chemical compounds. This study afforded valuable data to estimate the possible risk to health associated with pesticide exposure.     

The Frequency of Micronuclei among Newborns from Kragujevac, Central Serbia, after NATO Bombing in the Spring of 1999

Kragujevac, as an important industrial and economic center of the region, is now placed on the top of the list of environmentally endangered cities in our country. The aim of this biomonitoring study was to evaluate cytogenetic damage in a sample of newborns from Kragujevac after contamination of the environment entailed by the intensive bombing of the industrial zone of this city in the spring of 1999. The frequency of micronuclei (MN) in peripheral blood lymphocytes in a total of 61 phenotypically healthy newborns was analyzed before and after the NATO bombing. Analysis of micronuclei has been performed using the cytokinesis-block technique (CB MN test). Average MN frequency in lymphocytes of newborns before the bombing (N = 25) was 5.77 ± 0.85/1000 analyzed cells. After the bombing (N = 36), the average frequency of MN increased by a factor of 1.4 (8.11 ± 0.85), compared to the control frequency before the bombing. Statistical difference (p < 0.05) was established by Student's t-test. Our data suggest that such changes in genetic material were a direct consequence of contamination of the living environment.     

Genotoxic biomonitoring study of population residing in pesticide contaminated regions in Göksu Delta: micronucleus, chromosomal aberrations and sister chromatid exchanges

Pesticides are widely used throughout the world in agriculture to protect crops and in public health to control diseases. Nevertheless, exposure to pesticides represents a potential risk to humans. This paper describes a study of possible genetic damage in the people living in regions contaminated with complex mixture of pesticides in G?ksu Delta. In this study, used methods were chromosomal aberration (CA), sister chromatid exchange analysis (SCE) in the peripheral blood lymphocytes, and micronucleus (MN) assay in the buccal epithelial cells. In the present investigation, 32 affected subjects consist of 16 smoking and 16 non-smokings and an equal number of control subjects were assessed for genome damage. Micronucleus (MN), Broken egg (BE), Karyorrhexis (KR), Karyolysis (KL) and Binucleus (BN) frequencies were higher in affected subjects than in controls. Smoking had a statistically significant effect on the Micronucleus, Karyorrhexis and Binucleus frequencies for both the control and the exposed group. Also smoking and exposure affected the frequency of sister chromatid exchange and chromosomal aberrations compared with control groups.     

A global change of specialization and generalization in pollination networks from the plant perspective

A view of pollination systems is that they tend to generalization. Here we show that patterns exist in pollination systems that suggest otherwise. We examine how specialization may be affected by latitude, species richness and plant life-forms in 33 plant-pollinator interaction networks. Connectance increases with latitude (P = 0.040), but specialization at the community level significantly declines with increasing latitude, with the strongest evidence in the Northern Hemisphere (P = 0.001). The relationship between specialization level and connectance is negative (P < 0.001). While plants are more specialized in tropical areas and are more generalized at higher latitudes, specialization level also increases with plant richness (P < 0.001), total species richness (P = 0.041) and network size (P = 0.009). Specialization level of life forms of 1129 species differ substantially (P < 0.001), with herbs showing the highest specialization (74%), and tree species the lowest (62%).     

Genetic biomonitoring of an urban population exposed to mutagenic airborne pollutants

Biomonitoring studies have increased as a consequence of risks and effects to human health on exposure to environmental contaminants, mainly air pollutants. Genetic biomarkers are useful tools for the early assessment of exposure to occupational and environmental pollution. The objective of the present study was to investigate genotoxic effects on people residing and/or working downwind from an oil refinery in southern Brazil and the mutagenic activity of airborne particulate matter (PM10). Samples of peripheral blood and buccal mucosa cells were evaluated using the single-cell gel electrophoresis assay (comet assay) and the micronucleus (MN) assay, respectively. PM10 samples were collected in the target site and the organic matter extraced with dichloromethane was assessed for mutagenic activity in the Salmonella/microsome assay. The exposed group (n = 37) was compared to a reference group (n = 37) of subjects living in an urban area with limited traffic and industrial influence, located far from the main industrial areas. All PM10 organic extracts showed mutagenic positive responses and the effect decreased in the presence of S9 mix indicating that the predominant compounds present were direct-acting mutagens. The responses of YGs strains are consistent with aromatic amines and nitroarenes being present in the PM10 extracts. The group in the area under the influence of the oil refinery (exposed group) showed significantly higher DNA damage in lymphocytes than the reference group. The MN frequencies in buccal mucosa were very low for both groups and no difference between groups was observed. No association was found between age and tobacco smoking habit and level of DNA damages measured by the comet assay. The results indicate that the comet assay was a sensitive tool to detect DNA damage in subjects under the influence of an oil refinery, with marked genotoxic activity in the atmospheric environment.     

Monitoring DNA damage in <Emphasis Type="Italic">Mytilus galloprovincialis</Emphasis> from the Kaštela Bay in Croatia

The Kaštela Bay (Croatia) is known, by the use of Fast micromethod, as an area with the highest level of genotoxic agents along the Adriatic Sea. The genotoxic impacts in Kastela Bay and the neighbouring Trogir Bay using micronucleus test and Comet assay with mussel (Mytilus galloprovincialis) haemocytes were assesed during 2003 and 2004. This location received effluents from various industries (brewery, cement plant, etc.), ports (oil and general port), the Split shipyard, Vranjic shipworks along with domestic sewage and agricultural discharge that enters the bay without any treatment. In the meantime, a lot of industries were closed and the input of pollutants were discontinued. The tested mussels from the 2003–2004 confirmed that mussels from Kaštela Bay were affected by genotoxic contaminants. The aim of our study was to assess genotoxic impacts on Mytilus galloprovincialis in the same area after six years using Comet Assay test. Our survey showed decreses of genotoxic damage on mussel haemocytes.     

Cytogenetic monitoring of domestic mammals exposed to wastewaters from the localities of Dladla and Boukallou near Settat, Morocco

The aim of this work was to know the biological impact of wastewaters on livestock in the localities of Dladla and Boukallou (Town of Settat, Morocco), where wastewaters are largely used by the local population for their agricultural activities. We used the micronucleus (MN) test in cytochalasin B blocked binucleated cells (BNC) to assess the possible induced genetic effects. The cellular proliferation index (PI) was also calculated. Blood samples were collected from sheep, bovines and mules and peripheral blood cultures were made according to our laboratories' standard methodology. The results showed a significant increase of micronucleated cells in the lymphocytes of the exposed animals (14+/-8.79 MN/500 BNC) compared to the control (3+/-1.54 MN/500 BNC). The PI showed a nonsignificant decrease in the exposed animals. The highest MN frequency was found in mules which is probably due to their more sedentary life. Direct use of the wastewater by the animals as their source of drinking water significantly increased the frequency of cells with micronuclei. We also established an inverse correlation between the chromosome number and the PI. These results suggest a serious degradation of the ecosystem due to the wastewaters with a direct impact on its inhabitants.     

Internal distribution of uranium and associated genotoxic damages in the chronically exposed bivalve Corbicula fluminea

Uranium (U) internal distribution and involved effects in the bivalve Corbicula fluminea have been studied after direct chronic exposure (90 d, 10 μg.L-1). U distribution was assessed at the subcellular level (Metal Rich Granules -MRG-, pellets and cytosol fractions) in two main organs of the bivalve (gills and visceral mass). Micro-localisation was investigated by TEM-EDX analysis in the gills epithelium. DNA damage in gill and hemolymph samples was measured by the Comet assay. The 90-d exposure period led to a significant increase of U concentration in gills over time (×5) and a large U quantity in subcellular granules in gills. Finally, a significant increase (×2) in DNA damage was noted in exposed gills and haemocytes. This study shows that the accumulation levels and consequently the potential toxicity cannot be successfully predicted only on the basis of concentration in water or in tissues and subcellular fractions after chronic exposure.     

Bioaccumulation and biological effects in the earthworm Eisenia fetida exposed to natural and depleted uranium

The accumulations of both natural (U) and depleted (DU) uranium in the earthworms (Eisenia fetida) were studied to evaluate corresponding biological effects. Concentrations of metals in the experimental soil ranged from 1.86 to 600 mg kg⁻¹. Five biological endpoints: mortality, animals’ weight increasing, lysosomal membrane stability by measuring the neutral red retention time (the NRRT), histological changes and genetic effects (Comet assay) were used to evaluate biological effects in the earthworms after 7 and 28 days of exposure. No effects have been observed in terms of mortality or weight reduction. Cytotoxic and genetic effects were identified at quite low U concentrations. For some of these endpoints, in particular for genetic effects, the dose (U concentration)-effect relationships have been found to be non-linear. The results have also shown a statistically significant higher level of impact on the earthworms exposed to natural U compared to depleted U.     

Air pollution and young children's inhalation exposure to organophosphorus pesticide in an agricultural community in Japan

Assessment of airborne organophosphorus pesticides in houses of young children (1-6 years old) and childcare facilities was conducted following pesticide applications in an agricultural community in Japan. Trichlorfon and fenitrothion, applied in two separate periods, were frequently detected from outdoor and indoor air. Dichlorvos, the primary degradation product of trichlorfon, was also detected after the application of trichlorfon. Both the outdoors and indoor concentration of applied pesticide were shown to increase with decreasing distance from the pesticide-applied farm. Indoor concentration of these pesticides significantly correlated with outdoor concentration (p=0.001 for trichlorfon and p=0.001 for fenitrothion), indicating infiltration of applied pesticide inside. Ratio of indoor to outdoor concentration (I/O ratio) of fenitrothion was higher for houses with windows open during the application than those with closed windows (median value: 0.74 vs. 0.16, p=0.003). However, a similar trend was not observed for trichlorfon as well as dichlorvos in the first period. Dichlorvos was found to have a higher I/O ratio than trichlorfon during the period, and clear correlation between indoor concentrations of dichlorvos and those of trichlorfon suggested increased decomposition of trichlorfon in the indoor environment. Daily inhalation exposure estimated by using the fixed measurement data and time-activity questionnaire ranged from 0 to 35 ng/kg/day for trichlorfon, from 0 to 26 ng/kg/day for dichlorvos, and from 0 to 44 ng/kg/day for fenitrothion. Median inhalation exposure from indoor air accounted for 74%, 86.3%, and 45% of the daily inhalation exposure, respectively. For kindergarteners or nursery school children, inhalation exposure at childcare facilities was comparable with or more than that at home, indicating that pollution level at childcare facilities had potential of high impact on children's exposure. Estimated daily inhalation exposures were inversely correlated to the proximity of their activity location to the pesticide-applied farm.     

Genotoxic effect of exposure to metal(loid)s. A molecular epidemiology survey of populations living and working in Panasqueira mine area,Portugal

Previous studies investigating the exposure to metal(loid)s of populations living in the Panasqueira mine area of central Portugal found a higher internal dose of elements such as arsenic, chromium, lead, manganese, molybdenum and zinc in exposed individuals. The aims of the present study were to evaluate the extent of genotoxic damage caused by environmental and occupational exposure in individuals previously tested for metal(loid) levels in different biological matrices, and the possible modulating role of genetic polymorphisms involved in metabolism and DNA repair. T-cell receptor mutation assay, comet assay, micronucleus (MN) test and chromosomal aberrations (CA) were performed in a group of 122 subjects working in the Panasqueira mine or living in the same region. The modifying effect of polymorphisms in GSTA2, GSTM1, GSTP1, GSTT1, XRCC1, APEX1, MPG, MUTYH, OGG1, PARP1, PARP4, ERCC1, ERCC4, and ERCC5 genes was investigated. Significant increases in the frequency of all biomarkers investigated were found in exposed groups, however those environmentally exposed were generally higher. Significant influences of polymorphisms were observed for GSTM1 deletion and OGG1 rs1052133 on CA frequencies, APEX1 rs1130409 on DNA damage, ERCC1 rs3212986 on DNA damage and CA frequency, and ERCC4 rs1800067 on MN and CA frequencies. Our results show that the metal(loid) contamination in the Panasqueira mine area induced genotoxic damage both in individuals working in the mine or living in the area. The observed effects are closely associated to the internal exposure dose, and are more evident in susceptible genotypes. The urgent intervention of authorities is required to protect exposed populations.     

Assessment of adoption and impact of rainwater harvesting technologies on rural farm household income: the case of rainwater harvesting ponds in Rwanda

Rainwater harvesting is increasingly viewed as a major strategy for enhancing agricultural productivity and boosting farm income in many drought-prone areas. While this technology is being promoted in many developing countries, there is conflicting evidence in the literature about its impact on welfare of farm households. This study uses propensity score matching and discrete choice regression techniques to assess the impact of rainwater harvesting ponds on farm household income and factors that influence adoption of such technologies in Rwanda. It finds that households with rainwater harvesting ponds have significantly higher income than their counterparts of comparable observable characteristics. It further finds evidence that increase in farm income occurs via increased input use and that household size, asset endowments and participation in farmer organizations condition adoption of rainwater harvesting ponds. The study concludes that adoption of rainwater harvesting technologies has positive benefits on farm households. It discusses the policy implications that adoption of rainwater harvesting ponds presents a pathway for reducing rural poverty.     

Application of an improved method for the analysis of pesticides and their metabolites in the urine of farmer applicators and their families

As the annual use of pesticides in the United States has escalated, public health agencies have become increasingly concerned about chronic pesticide exposure. However, without reliable, accurate analytical methods for biological monitoring, low-level chronic exposures are often difficult to assess. A method for measuring simultaneously the urinary residues of as many as 20 pesticides has been significantly improved. The method uses a sample preparation which includes enzyme digestion, extraction, and chemical derivatization of the analytes. The derivatized analytes are measured by using gas chromatography coupled with isotope-dilution tandem mass spectrometry. The limits of detection of the modified method are in the high pg/L - low μg/L range, and the average coefficient of variation (CV) of the method was below 20% for most analytes, with approximately 100% accuracy in quantification. This method was used to measure the internal doses of pesticides among selected farmer applicators and their families. Definite exposure and elimination patterns (i.e., an increase in urinary analyte levels following application and then a gradual decrease to background levels) were observed among the farmer applicators and many of the family members whose crops were treated with carbaryl, dicamba, and 2,4-D esters and amines. Although the spouses of farm workers sometimes exhibited the same elimination pattern, the levels of the targeted pesticides or metabolites found in their urine were not outside the ranges found in the general U.S. population (reference range). The farmer applicators who applied the pesticides and some of their children appeared to have higher pesticide or metabolite levels in their urine than those found in the general U.S. population, but their levels were generally comparable to or lower than reported levels in other occupationally exposed individuals. These results, however, were obtained from a nonrandom sampling of farm residents specifically targeted to particular exposures who may have altered their practices because they were being observed; therefore, further study is required to determine if these results are representative of pesticide levels among residents on all farms where these pesticides are applied using the same application techniques. Using this method to measure exposure in a small nonrandom farm population allowed differentiation between overt and background exposure. In addition, the important role of reference-range information in distinguishing between various levels of environmental exposure was reaffirmed.     

Dioxin and dioxin-like PCB profiles in the serum of industrial and municipal waste incinerator workers in Korea

To assess occupational exposure, we determined the concentrations of PCDD/Fs and PCBs in human serum samples from 26 incinerator workers (10 industrial waste and 16 municipal solid waste incinerator workers), 38 residents near the facilities and 7 inhabitants (as control subjects) living over 10 km away from any incinerator facilities in Korea. The mean TEQ_WHO levels of PCDD/Fs in the industrial and MSWI workers were 41.57 and 9.86 pg TEQ_WHO g^? 1 lipid, respectively. For the residents, the mean TEQ_WHO was 13.47 pg TEQ_WHO g^? 1 lipid (residents near IWI, MSWI, and control subjects: 17.64, 13.31, and 6.91 pg TEQ_WHO g^? 1 lipid). Higher levels of certain PCDD/F congeners, mainly PCDFs, were detected in the serum of industrial incinerator workers in comparison to the levels measured in the residents. Significant differences were observed for PCDFs, the major compounds were OCDF, 1,2,3,4,6,7,8-HpCDF, and the minor components 1,2,3,4,7,8-HxCDF, 1,2,3,6,7,8-HxCDF, and 2,3,4,6,7,8-HxCDF (p < 0.01). The PCDD/F congener patterns and concentrations measured in the 71 serum samples examined suggest that the industrial incinerator workers were exposed to PCDD/Fs in the workplace, possibly through inhalation and/or skin contact. In contrast, the levels and congener patterns of PCBs measured were similar in all subjects, indicating that workers from the incinerator facilities examined were not subjected to additional exposure to these compounds.     

Can fine particulate matter explain the paradoxical ozone associations?

Our previous paper entitled "Paradoxical ozone associations could be due to methyl nitrite from combustion of methyl ethers or esters in engine fuels" (Env. Int.. 2007;33;1090) reviewed 11 studies of the impact of ozone on human health that, paradoxically, found a negative coefficient for ozone-morbidity associations. We argued that the most likely explanation for this effect would be methyl nitrite (MN) as an unsuspected exhaust component of engines with methyl ether in the fuel. The basis of the argument was the fact that MN is rapidly destroyed by sunlight, so that MN would be negatively correlated with ozone. All (but one) of the reviewed studies concluded that criterion pollutants could not explain the negative slope. The argument was strengthened by the observation that such paradoxical ozone associations have not been found in regions without significant methyl ether in gasoline. Left unaddressed in the previous paper was the possibility that fine particulate matter (FPM) might explain the POA. If this were true, then it would be necessary that the FPM be negatively correlated with ozone in those regions that found a POA. The current paper reviews data on FPM-ozone correlations in those regions where a POA was identified. The results show that FPM was, in most cases, positively correlated with ozone and so could not explain the POA.     

Investigating the embryo/larval toxic and genotoxic effects of γ irradiation on zebrafish eggs

Eggs/larval of freshwater fish (Danio rerio) were exposed to low dose rates of external gamma radiation (from 1 to 1000 mGy d⁻¹) over a 20-day period, with the objective of testing the appropriateness of the 10 mGy d⁻¹ guideline suggested by the IAEA. The present study examines different endpoints, mortality and hatching time and success of embryos as well as the genotoxicity of γ-irradiations (after 48 h). The 20-day embryo-larval bioassay showed an enhanced larval resistance to starvation after chronic exposure to γ irradiation (from low 1 mGy d⁻¹ to high dose rate 1000 mGy d⁻¹) and an acceleration in hatching time. Gamma irradiation led to increased genotoxic damage Ito zebrafish egg (40-50% DNA in tail in Comet assay) from the lowest dose rate (1 mGy d⁻¹). Possible mechanisms of γ radiotoxicity and implications for radioprotection are discussed.     

城乡交错带土地利用方式对土壤理化性质的影响

以长沙市南郊山地丘陵区为研究对象,选择7种不同土地利用方式（次生林地、经济林地、杉木人工林、采伐迹地、撂荒地、苗圃地、坡耕地）研究了城乡交错带不同土地利用方式对土壤理化性质的影响。结果表明：7种土地利用方式下,土壤颗粒组成主要集中在1～0.05 mm和＜0.001 mm两个粒级范围内,土壤质地以重壤土和轻粘土为主。随着人为干扰程度的减弱,土壤砂粒（1.0～0.05 mm）、土壤粉粒（0.05～0.01 mm）百分率逐渐下降,＜0.01 mm的土壤物理性粘粒百分率增加。自然、半自然状态利用方式的林地（次生林地、经济林地、杉木人工林地）土壤总孔隙度、含水量普遍较高,土壤密度低于坡耕地、苗圃地、撂荒地、采伐迹地,而土壤pH值、有机质含量及氮、磷、钾含量普遍高于坡耕地、苗圃地、撂荒地、采伐迹地