Histopathological alterations in gill, liver and kidney of common carp exposed to chlorpyrifos

Histopathological alterations in gill, liver and kidney of common carp, Cyprinus carpio, intoxicated with sub-lethal concentrations of chlorpyrifos (O,O,-diethyl-O-3,5,6-trichloro-2-pyridyl phosphorothioate) pesticide (1 and 100 μg/L) for a period of 14 days were analyzed under light microscope. Gill exhibited hyperplasia and hypertrophy of gill epithelium, blood congestion, dilation of marginal channel, epithelial lifting, lamellar fusion, lamellar disorganization, lamellar aneurysm, rupture of the lamellar epithelium, rupture of pillar cells and necrosis. Alterations in hepatocytes were more pronounced, including nuclear and cellular hypertrophy, cellular atrophy, irregular contour of cells and nucleus, cytoplasmic vacuolation, cytoplasmic and nuclear degeneration, cellular rupture, pyknotic nucleus, necrosis and melanomacrophages aggregations. Histopathological lesions in kidney were cellular and nuclear hypertrophy, narrowing of tubular lumen, cytoplasmic vacuolation, hyaline droplet degeneration, nuclear degeneration, occlusion of tubular lumen, tubular regeneration, dilation of glomerular capillaries, degeneration of glomerulus and hemorrhage in Bowman's space. The most significant conclusion drawn from this study was that with the increased concentration and duration the toxicosis of chlorpyrifos would be enhanced as shown through the analysis of mean assessment value (MAV) and degree of tissue changes (DTC) also.     

Histopathological changes and antioxidant response in brain and kidney of common carp exposed to atrazine and chlorpyrifos

We investigated oxidative stress response and histopathological changes in the brain and kidney of the common carp after a 40-d exposure to CPF and ATR, alone or in combination, and a 20-d recovery. Superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) activities and malondialdehyde (MDA) content were measured using standard assays. Our results indicated that exposure to ATR, CPF or a combination of the two for 40 d induced significant changes in antioxidant enzyme (SOD, CAT and GSH-Px) activities and MDA content in the brain and kidney of the common carp. Pathological changes included tissue damage that was more severe with increased of exposure dose. To our knowledge, this is the first report to study oxidative stress and histopathological effects caused by subchronic exposure to ATR, CPF and ATR/CPF combination on common carp. The information presented in this study may be helpful to understanding the mechanisms of ATR-, CPF- and ATR/CPF combination-induced oxidative stress in fish.     

Toxic effects of chlorpyrifos on lysozyme activities,the contents of complement C3 and IgM,and IgM and complement C3 expressions in common carp (Cyprinus carpio L.)

Chlorpyrifos is one of the organophosphate pesticides widely used in agricultural practices throughout world. It has resulted in a series of toxicological and environmental problems, such as impacts on many non-target aquatic species, including fish. In the present study, toxic effects of chlorpyrifos on lysozyme activities, contents of IgM and complement C3, and the expressions of IgM and complement C3 at mRNA level in common carp were evaluated by acute exposure of 15 (1/10 LC₅₀) or 75 μg L⁻¹ (1/2 LC₅₀) of chlorpyrifos for 7 d. The results of acute toxicity tests showed that the 96 h-LC₅₀ of chlorpyrifos for common carp was determined to be 149 μg L⁻¹. We also found that chlorpyrifos promoted lysozyme activities at the earlier stages of exposure but inhibited it at the late stages in the serum, hepatopancreas, and spleen of common carp. Furthermore, it was observed that chlorpyrifos-exposure decreased IgM contents in fish serum and spleen while increased it in kidney. No obvious change was found in the contents of complement C3 in fish spleen, while a slight increase of complement C3 was observed in fish serum and kidney after 1 d of chlorpyrifos-exposure. In addition, the results of quantitative real-time PCR showed that IgM and complement C3 expressions were up-regulated at the earlier stage of exposure but down-regulated at later stage. Our results indicate that chlorpyrifos causes immunotoxicity to common carp.     

Reference values for acetyl and butyrylcholinesterases in cattle under actual management conditions,hepatic and renal function by application of chlorpyrifos

Chlorpyrifos is an anticholinesterase organophosphate insecticide widely used in Argentina in the production of food derived from animal, fruit and horticultural origin and is reported as a residue within these products. Local reference values for acetyl and butyrylcholinesterase were determined in Aberdeen Angus bovine and cross bred cattle (n = 25), a requirement to be able to evaluate toxicity of commercial organophosphate and carbamate formulations. The activity of cholinesterase enzymes presented an overall mean of 2,183.00 ± 485.6 IU L^?1 for erythrocyte acetylcholinesterase and 203.1 ± 42.06 IU L^?1 for plasma butyrylcholinesterase, which are used as reference values for meat steers within a system of intensive production in a semi-arid region. The toxic potential of chlorpyrifos in steers of the same breeds (n = 12) was assessed applying chlorpyrifos 15.00% Tipertox® in a single therapeutic dose of 7.50 mg kg^?1 by topical route. Prior to application and then on day 1 and day 21 post-application, both blood cholinesterases, serum chlorpyrifos concentration by ultra-high resolution liquid chromatography with mass detector, analysis of blood counts, total proteins, liver enzymes, urea and creatinine were evaluated. The mean plasma concentration of chlorpyrifos was 27.90 ug L^?1 at 24 h. The findings indicate that the therapeutic treatment of castrated male bovines treated with chlorpyrifos, applied by pour-on according to the manufacturer's instructions, does not cause changes in the variables evaluated.     

Endosulfan acute toxicity and histomorphological alterations in Jenynsia multidentata (Anablepidae,Cyprinodontiformes)

Toxicity tests using adult specimens of Jenynsia multidentata were carried out during 96 hours in order to determine the lethal concentration (LC50) of endosulfan. Histological alterations were determined in gills and liver. Gill damage was quantified as secondary lamellae thickness. The 96 hr LC50 values were significantly different between males (0.719 μ g.L^{? 1}) and females (1.317 μg.L^{? 1}). The sex difference was attributed to the dimorphism in the lipid content in females (2.16%) and males (1.79%). Histological alterations in gills included hypertrophy and lifting of the epithelium of the secondary lamellae and aneurisms. These alterations caused a significant increase of the secondary lamellae thickness in treatment versus control fish. Finally, reversible histological alterations (such as hydropic degeneration and dilation of sinusoids) were observed in the liver of exposed fish as well as an irreversible change such as necrosis at the highest concentrations.     

Oxidative stress,hematological and biochemical alterations in farmers exposed to pesticides

In this study, a cohort of farmers from the Mateur region in the North of Tunisia, were interviewed and examined for the biochemical effects of pesticides. We studied their haematological profile, lipid parameters, serum markers of nephrotoxicity and hepatotoxicity. We also evaluated the activities of Butyrylcholinesterase (BChE), Acetylcholinesterase (AChE) and thiolactonase-paroxonase (PON). Moreover, lipid peroxidation and activities of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) were determined. The duration of pesticide use and the farmers’ age were considered in the analysis. Our results revealed significant differences in some haematological parameters, in liver and kidney functions, in the lipidic status of the pesticide-exposed group. We also reported an increase in the index of incidence of cardiovascular risk in farmer populations. A significant decrease in AChE, BChE and PON levels was found among farmers. Lipid peroxidation, however, increased. The activities of SOD and CAT were remarkably elevated in farmer populations. There was a significant relation between changes in biological markers, the duration of pesticide use and the farmers’ age. This study indicates that a long-term exposure to pesticides may play an important role in the development of vascular diseases via metabolic disorders of lipoproteins, lipid peroxidation and oxidative stress, inhibition of BChE and decrease in thiolactonase-PON levels.     

Hydrolysis of di-n-butyl phthalate, butylbenzyl phthalate and di(2-ethylhexyl) phthalate in human liver microsomes

Diester phthalates are industrial chemicals used primarily as plasticizers to import flexibility to polyvinylchloride plastics. In this study, we examined the hydrolysis of di-n-butyl phthalate (DBP), butylbenzyl phthalate (BBzP) and di(2-ethylhexyl) phthalate (DEHP) in human liver microsomes. These diester phthalates were hydrolyzed to monoester phthalates (mono-n-butyl phthalate (MBP) from DBP, mono-n-butyl phthalate (MBP) and monobenzyl phthalate (MBzP) from BBzP, and mono(2-ethylhexyl) phthalate (MEHP)) by human liver microsomes. DBP, BBzP and DEHP hydrolysis showed sigmoidal kinetics in V-[S] plots, and the Hill coefficient (n) ranged 1.37-1.96. The S₅₀, V_max and CL_max values for DBP hydrolysis to MBP were 99.7 μM, 17.2 nmol min⁻¹ mg⁻¹ protein and 85.6 μL min⁻¹ mg⁻¹ protein, respectively. In BBzP hydrolysis, the values of S₅₀ (71.7 μM), V_max (13.0 nmol min⁻¹ mg⁻¹ protein) and CL_max (91.3 μL min⁻¹ mg⁻¹ protein) for MBzP formation were comparable to those of DBP hydrolysis. Although the S₅₀ value for MBP formation was comparable to that of MBzP formation, the V_max and CL_max values were markedly lower (<3%) than those for MBzP formation. The S₅₀, V_max and CL_max values for DEHP hydrolysis were 8.40 μM, 0.43 nmol min⁻¹ mg⁻¹ protein and 27.5 μL min⁻¹ mg⁻¹ protein, respectively. The S₅₀ value was about 10% of DBP and BBzP hydrolysis, and the V_max value was also markedly lower (<3%) than those for DBP hydrolysis and MBzP formation for BBzP hydrolysis. The ranking order of CL_max values for monoester phthalate formation in DBP, BBzP and DEHP hydrolysis was BBzP to MBzP ? DBP to MBP > DEHP to MEHP > BBzP to MBP. These findings suggest that the hydrolysis activities of diester phthalates by human liver microsomes depend on the chemical structure, and that the metabolism profile may relate to diester phthalate toxicities, such as hormone disruption and reproductive effects.     

Hematological and histopathological changes in silver catfish Rhamdia quelen (Siluriformes) exposed to clomazone herbicide in the Madre River,Santa Catarina State,Southern Brazil

This study evaluated the influence of the clomazone herbicide (2-[(2-chlorophenyl)methyl]-4,4-dimethyl-3-isoxazolidinone) contamination on the hematological parameters and histological changes in gills and liver of silver catfish (Rhamdia quelen) from Madre River, Santa Catarina State, Southern Brazil. Fish were collected between March 2010 and January 2012 at two different sites of the Madre River, one site receiving residual water (contaminated site) from rice culture (n = 49) and another that do not receive residual water (reference site) (n = 48). The herbicide clomazone analysis detected 3.40 ± 1.70 μg/L in the contaminated site and 1.1 ± 0.33 μg/L in the reference site. Fish from contaminated site showed increased (P < 0.05) number of monocytes suggesting the possible defense response as a result of chronic exposure to clomazone. On the other hand, no difference was found in the hematocrit percentage, red blood cell count, total thrombocyte number, white blood cell count, lymphocytes, and neutrophils number. Fish from both sites showed histopathological changes in gills and liver, possibly caused by chronic exposure to contamination. The influence of herbicide sub doses on fish health is also discussed.     

斯里兰卡CKDu病区地下水源饮用水关键问题及解决策略

不明原因肾病(CKDu)是斯里兰卡亟需解决的重要民生问题,其地下水源饮用水中的溶解性污染物可能是重要的致病原因。系统分析了北部中心省内高、低、非发病区及对照区 (HR、LR、NR、CR)干湿季地下水的水化学特征和DOM特性,结果表明：地下水主要呈Ca-Mg-HCO₃型,硬度和F⁻超标率较高,是地下水作为饮水安全保障最关键的指标;地下水中DOM主要为自生源,芳香性和腐熟化程度较低,主要为富里酸、腐殖酸和小分子蛋白质类物质;湿季DOM浓度较高,是Fe浓度增加的关键原因;而干季地下水中微生物活性较高,腐殖质和蛋白质发生分解,小分子络氨酸/赖氨酸的浓度增加。同时,发现不同区域地下水中DOM的生物利用性表现为HR−1,芳香性较高。有机无机的相互作用导致赋存形态、生物可利用性的变化可能是CKDu致病的重要原因。针对以上问题,提出“吸附-除F-软化-NF-消毒”的处理策略,通过有机物吸附处理、阴离子交换除F、软化除硬分离预处理及NF深度处理等技术有效解决地下水中DOM、F⁻、Ca²⁺和Mg²⁺超标的问题,实现饮用水的供给。本研究全面分析了CKDu病区地下水中的污染物组成,并提出解决策略,可为斯里兰卡饮用水安全保障提供支撑。     

Distribution of pesticides and bisphenol A in sediments collected from rivers adjacent to coral reefs

To investigate the deteriorating health of coral reefs in Okinawa, Japan, natural sediment samples were analyzed for diuron, Irgarol 1051, chlorpyrifos, and bisphenol A (BPA) which are hazardous to corals. Samples were analyzed by solid-phase extraction (SPE) followed by high-performance liquid chromatography with tandem mass spectrometry (LC–MS–MS). Although diuron and chlorpyrifos usage is only well recorded for farms and not for cities, these chemicals were detected in both rural and urban areas. Additionally, diuron concentration in urban areas was in some cases higher than in rural areas, which might be caused by greater consumption of these chemicals in home gardens in city areas. Irgarol 1051 was detected in downstream river areas, which are situated far from the source sites such as pier or fishery harbor (0.6–3.2 km). This result suggested that Irgarol 1051 could be transported from the river mouths to the sampling sites during flood tides. High BPA concentrations were associated with urban areas (<1.2–22.0 μg kg⁻¹), while low concentrations were associated with rural areas (nd–6.8 μg kg⁻¹). The river sediments under study are delivered to coral reefs in large quantity through runoff caused by typhoons and other heavy rains. The highly hazardous chemicals are carried into coral reefs on these sediments. Therefore, these hazardous chemical substances may already be influencing the coral reefs.     

Endocrine modulation, inhibition of ovarian development and hepatic alterations in rainbow trout exposed to polluted river water

Under laboratory conditions, female rainbow trout were exposed to graded concentrations of water from the River Lambro, a polluted tributary of the River Po, and to the effluent of a large wastewater treatment plant which flows into the River Lambro. In field exposures, trout were held in cages in the River Po upstream and downstream from the confluence of the River Lambro. After 10-day (laboratory) and 30-day (laboratory and field) exposures, trout were examined for several chemical, biochemical and histological endpoints. The results indicated that exposure to complex mixtures of chemicals, including estrogen receptor agonists, aryl-hydrocarbon receptor agonists, and probably antiandrogens, had occurred. Exposure altered the plasma levels of 17β-estradiol and testosterone, and some treatments also enhanced the activity of hepatic ethoxyresorufin O-deethylase. Gonadal histology showed varying levels of degenerative processes characterised by oocyte atresia, haemorrhages, melano-macrophage centres (MMCs), and oogonia proliferation. Liver histology showed less severe effects.     

河蚬对太湖梅梁湾沉积物中HCHs和DDTs的生物富集

以采自清洁水体的河蚬(Corbicula fluminea)为实验生物,利用生物富集实验测试了河蚬对太湖梅梁湾水源地沉积物中HCHs和DDTs的生物富集.研究结果表明,将河蚬暴露于有机氯农药污染程度相似的太湖梅梁湾沉积物中(HCHs和DDTs浓度分别为1.5-1.8.S/S(以干重计)和1.1-1.7 as/g(以干重计))后,随着暴露时间的延长(24～168h),河蚬对有机氯农药的富集量随着暴露时间逐渐增加.试验结束时各样点中河蚬对HCHs和DDTs的富集量分别为(9.4±2.2)ng/g(以干重计)、(20.7±7.6)ng/g(以干重计).实验水体沉积物中HCHs和DDTs的生物沉积物富集因子(BSAF)分别为1.5±0.1和4.4±0.7.生物-沉积物富集因子(BSAF)与有机物的辛醇-水比值(Kow)存在显著正相关.     

Detection of Class I and II integrons for the assessment of antibiotic and multidrug resistance among Escherichia coli isolates from agricultural irrigation waters in Bulacan,Philippines

Contaminated irrigation water may greatly affect not only the quality of produce but also the people exposed to it. In this study, agricultural irrigation waters in Bulacan, Philippines were assessed and found to be contaminated with Escherichia coli (E. coli) ranging from 0.58 to 4.51 log₁₀ CFU/mL. A total of 79 isolates of E. coli were confirmed through polymerase chain reaction (PCR) amplifying the uidA gene and were tested for phenotypic resistance using 10 antimicrobials through the Kirby–Bauer disc diffusion method. Forty-six isolates (58.22%) were noted to be multidrug resistant (MDR) with high resistance rate to cephalothin, tetracycline, streptomycin, ampicillin, trimethoprim, nalidixic acid, and chloramphenicol. Moreover, this study also examined the prevalence of Class I and II integrons accounting to 67.39% and 17.39%, respectively, of the MDR E. coli strains using multiplex PCR. The results imply that the agricultural water used in Bulacan is contaminated with the fecal material of man or other animals present in the area, and the presence of MDR bacteria, which pose a potential threat to individuals in these areas, is alarming. In addition, detection of integrons could be a good marker for the identification of MDR isolates. Lastly, this study could develop strategies for the proper management of farming sites leading to the detection of food-borne pathogens and prevention of infectious diseases.     

Combined application of conservative transport modelling and compound-specific carbon isotope analyses to assess in situ attenuation of benzene, toluene, and o-xylene

In recent years, compound specific isotope analyses (CSIA) have developed into one of the most powerful tools for the quantification of in situ biodegradation of organic contaminants. In this approach, the calculation of the extent of biodegradation of organic contaminants in aquifers is usually based on the Rayleigh equation, and thus neglects physical transport processes such as dispersion that contribute to contaminant dilution in aquifers. Here we combine compound specific isotope analyses with a conservative transport model to study the attenuation of aromatic hydrocarbons at a former gasworks site. The conservative transport model was first used to simulate concentration reductions caused by dilution at wells downgradient of a BTEX source. In a second step, the diluted concentrations, together with the available stable carbon isotope ratios and carbon fractionation factors for benzene, toluene and o-xylene were applied in the Rayleigh equation to quantify the degree of biodegradation at each of those wells. At the investigated site, where other attenuation processes such as sorption and volatilisation were proven to be negligible, the combined approach is recommended for benzene, which represents a compound for which the effect of biodegradation is comparable to or less than the effect of dilution. As demonstrated for toluene and o-xylene, the application of the Rayleigh equation alone is sufficient if dilution can be proved to be insignificant in comparison to biodegradation. The analysis also suggests that the source width and the position of the observation wells relative to the plume center line are significantly related to the degree of dilution.     

Assessment of oxidative stress due to exposure to pesticides in plasma and urine of traditional limited-resource farm workers: Formation of the DNA-adduct 8-hydroxy-2-deoxy-guanosine (8-OHdG)

The objective of this study was to assess the risk of genotoxicity caused due to oxidative stress using plasma and urinary levels of 8-hydroxy-2-deoxyguanosine (8-OHdG) in farm workers for six months during a growing season. Blood and urine samples were collected once a month for six months (June to November 2003) from farm workers (n = 15) and urban unexposed controls (n = 10). Plasma and urinary levels of 8-OHdG were evaluated by Enzyme Linked Immunosorbent Assay (ELISA) technique. There was no significant difference in the urinary levels of 8-OHdG between the farm workers and the control group, but there was an approximately four-fold increase in mean values of plasma 8-OHdG levels in the farm worker group (P ≤ 0.05).     

Correlations between mutagenic activity of organic extracts of airborne particulate matter, NOx and sulphur dioxide in southern Germany: results of a two-year study

Goals, Scope and Background Among other substances, sulphur dioxide (SO2), nitric oxide (NO) and nitrogen dioxide (NO2) are parameters which are routinely measured to describe basic air quality. Organic extracts of airborne particulate matter contain mutagenic chemical compounds of different origins. The aim of the study was to find correlations between routine monitoring data and mutagenic activity of organic extracts of simultaneously drawn samples.Methods Specimens were collected over a period of two years at 8 sampling sites in south-west Germany. Simultaneously, concentrations of NO, NO2, and SO2 were measured on-line within the framework of the official air monitoring network of Baden-Württemberg, Germany. Dust samples were collected for biotesting using high volume air samplers equipped with glass fibre filters. After sampling was completed, filters were extracted and samples were prepared for biological testing. Mutagenic activity was tested by means of the plate incorporation assay (Ames test) using S. typhimurium TA98 and TA100 tester strains. During the first year of the study, all tests have been performed with and without metabolic activation. Additionally, a series of tests has been performed in parallel with TA98 and TA98NR.Results and Discussion Comparison of Ames test data obtained with and without metabolic activation indicates no statistically significant difference between both methods. Therefore, during the second year of the study, all tests have been performed without metabolic activation. Average yearly activities at the sampling sites were between 1 und 27 Revertants per m³ (Rev/m³). High activities were preferably found at congested sites (Karlsruhe, up to 95 Rev/m³). However, peak values of over 100 Rev/m³ were found in other places where pollution by traffic is significantly lower. The reason for these high level values is not evident. Tests performed using TA98NR tester strain indicate a significant share (average 31%) of compounds requiring activation by nitroreductase for mutagenic activity. Average mutagenic activity can be correlated to routine monitoring parameters. Comparison of averaged data for particular sampling sites indicates significant correlation between nitric oxide and mutagenic activity in TA98 (r²=0.90), while correlation between nitrogen dioxide (0.84) or sulphur dioxide (0.52) and mutagenic activity is weaker. For TA100, correlations are generally weaker than for TA98. Comparison of data for mutagenic activity and routine monitoring data of distant sites being sampled simultaneously shows parallel behaviour.Conclusions Results from this study show that mutagenic activity can be compared to seasonal and local variations of gaseous indicator air pollutants. Tester strain TA98 generally shows the best correlations. Although pollution by particle-bound mutagenic substances is significantly higher during the cold season than during summer on average, mutagenic activity of airborne dust is not a continuous effect. During winter, peak levels as well as low pollution periods can occur. Even during winter time mutagenic activity can reach very low levels typical for summertime. Comparison of results for distant sampling sites where samples have been collected simultaneously indicate that “classical” indicators of air pollution and bacterial mutagenicity of organic extracts from airborne particulate matter are influenced by connected effects. Seasonal trend of mutagenic activity, in particular, is similar to the concentrations of nitrogen oxide. NO is a strong indicator for vehicle exhaust gases. It is concluded that the average mutagenic activity at particular sites can be estimated using NO concentrations as an indicator.     

Contribution of motor vehicle emissions to organic carbon and fine particle mass in Pittsburgh, Pennsylvania: Effects of varying source profiles and seasonal trends in ambient marker concentrations

We present estimates of the vehicular contribution to ambient organic carbon (OC) and fine particle mass (PM) in Pittsburgh, PA using the chemical mass balance (CMB) model and a large dataset of ambient molecular marker concentrations. Source profiles for CMB analysis are selected using a method of comparing the ambient ratios of marker species with published profiles for gasoline and diesel vehicle emissions. The ambient wintertime data cluster on a hopanes/EC ratio–ratio plot, and therefore can be explained by a large number of different source profile combinations. In contrast, the widely varying summer ambient ratios can be explained by a more limited number of source profile combinations. We present results for a number of different CMB scenarios, all of which perform well on the different statistical tests used to establish the quality of a CMB solution. The results illustrate how CMB estimates depend critically on the marker-to-OC and marker-to-PM ratios of the source profiles. The vehicular contribution in the winter is bounded between 13% and 20% of the ambient OC (274±56–416±72 ng-C m⁻³). However, variability in the diesel profiles creates uncertainty in the gasoline–diesel split. On an OC basis, one set of scenarios suggests gasoline dominance, while a second set indicates a more even split. On a PM basis, all solutions indicate a diesel-dominated split. The summer CMB solutions do not present a consistent picture given the seasonal shift and wide variation in the ambient hopanes-to-EC ratios relative to the source profiles. If one set of source profiles is applied to the entire dataset, gasoline vehicles dominate vehicular OC in the winter but diesel dominates in the summer. The seasonal pattern in the ambient hopanes-to-EC ratios may be caused by photochemical decay of hopanes in the summer or by seasonal changes in vehicle emission profiles.