Copper induces damage, oxidative stress and cell death in endothelium of chronic intoxicated Wistar rats / El cobre induce daño, estrés oxidativo y muerte celular en el endotelio de ratas Wistar intoxicadas

SUMMARY: Reactive Oxygen Species (ROS) are part of the functional balance of various systems, they can generate cellular damage by oxidative stress associated with disease processes such as atherosclerosis, cardiovascular disease, diabetes, and aging. Some studies report that copper induces damage to the endothelium, which could be associated with cardiovascular pathologies. This study was an experimental comparative, prospective, longitudinal, and controlled clinical trial in a murine animal model. Twenty-four male Wistar rats were included, the distribution of the groups was time-depending chronic exposition to copper, and a control group. Results show gradual alterations in the groups treated with copper: areas with loss of the endothelium, signs of disorganization of smooth muscle fibers in the tunica media, as well as areas with the fragmentation of the elastic sheets. A significant statistical difference was observed in the active- Caspase-3 analysis expression in the aortic endothelium and endothelium of the capillaries and arterioles of the lung between the control group vs 300 ppm of copper. Expression of eNOS was detected in the endothelium of the aorta and vessels of the lung. Our study shows histological changes in the walls of the great vessels of intoxicated rats with copper, and the increment of inflammatory cells in the alveoli of the study model, mainly at a high dose of copper exposition. These results will be useful to understand more about the mediators involved in the effect of copper over endothelium and cardiovascular diseases in chronic intoxication in humans.

RESUMEN: Las Especies Reactivas de Oxígeno (ROS) son parte del equilibrio funcional de varios sistemas, pueden generar daño celular por estrés oxidativo asociado a procesos patológicos como aterosclerosis, enfermedades cardiovasculares, diabetes y envejecimiento. Algunos estudios informan que el cobre induce daños en el endotelio, lo que podría estar asociado a patologías cardiovasculares. Este estudio fue un ensayo clínico experimental comparativo, prospectivo, longitudinal y controlado en un modelo animal murino. Se incluyeron veinticuatro ratas Wistar macho, la distribución de los grupos fue la exposición crónica al cobre en función del tiempo y un grupo de control. Los resultados muestran alteraciones graduales en los grupos tratados con cobre: áreas con pérdida del endotelio, signos de desorganización de las fibras musculares lisas en la túnica media, así como áreas con la fragmentación de las láminas elásticas. Se observó una diferencia estadística significativa en la expresión del análisis de caspasa-3 activa en el endotelio aórtico y el endotelio de los capilares y arteriolas del pulmón entre el grupo de control frente a 300 ppm de cobre. Se detectó expresión de eNOS en el endotelio de la aorta y los vasos del pulmón. Nuestro estudio muestra cambios histológicos en las paredes de los grandes vasos de ratas intoxicadas con cobre, y el incremento de células inflamatorias en los alvéolos del modelo de estudio, principalmente a una alta dosis de exposición de cobre. Estos resultados serán útiles para comprender más sobre los mediadores involucrados en el efecto del cobre sobre el endotelio y las enfermedades cardiovasculares en la intoxicación crónica en humanos.

Copper formulations in bacterial blight control and toxic effects on coffee seedlings

Bacterial blight of coffee (Pseudomonas syringae pv. garcae) is an important coffee disease and can be controlled using antibiotics and copper-based compounds. However, copper-based compounds raise doubts among coffee growers regarding bacterial blight control efficiency and phytotoxic potential. In this work, coffee plants were sprayed with different copper molecules in order to study their efficiency on bacterial blight control and the phytotoxic potential. Seven copper formulations, cuprous oxide, copper oxychloride, copper nitrate, copper hydroxide 1 (water-dispersible granules) and 2 (concentrated suspension), copper sulfate 1 (complexed with gluconic acid) and 2 (Bordeaux mixture) were studied. The copper formulations efficiency was compared with the antibiotic kasugamycin, saline solution, and control. In controlled environmental conditions of temperature, relative humidity, and photoperiod, coffee seedlings were sprayed with the treatments and after 24 hours they were inoculated with Pseudomonas syringae pv. garcae suspension. Disease incidence and severity assessments were performed in a 2-day interval during a 16-day period. Phytotoxicity incidence and severity, mapping, and quantification of copper on the leaf tissue surface, dried leaves weight, and total copper leaf content were assessed 16 days after pathogen inoculation. Data were submitted to the Scott-Knott test (p < 0.05). Cuprous oxide and copper sulfate 2 proved most efficient to bacterial blight control, causing lower phytotoxicity effect, best covering, and persistence on leaf tissues. Copper nitrate and copper sulfate complexed with gluconic acid were more phytotoxicity compared to other copper formulations.

Avaliação da contaminação por cobre e chumbo do lago Aratimbó - Umuarama - Pr / Evaluation of copper and lead contamination of the Aratimbó lake in Umuarama - Paraná

Este trabalho teve como objetivo quantificar os metais Cobre e Chumbo presentes na água e no sedimento do lago Aratimbó, localizado na cidade de Umuarama-PR e analisar os indicadores abióticos de temperatura e pH da água. As coletas foram feitas em triplicatas, em três pontos distintos, em dois diferentes dias, sendo o primeiro em tempo seco, anterior a chuva e outro logo após a primeira chuva. As amostras tiveram sua temperatura e pH verificado in loco e a determinação dos metais ocorreu por meio de Espectrofotometria de Absorção Atômica por chama. Os resultados de pH obtiveram valores satisfatórios em todos os pontos. Em relação à concentração de metais, na água, houve maior concentração de chumbo antes da chuva e uma concentração abaixo do limite apenas para o cobre no ponto 1. No sedimento, a concentração de ambos metais foi maior antes da chuva, exceto no ponto três, onde a concentração foi maior após a chuva. É possível que os maiores valores de concentração para os metais tenham sido superiores no período de seca em virtude do volume aumentado de água após a chuva, com maior diluição. Assim, podendo atribuir a contaminação às atividades antropogênicas, uma vez que o lago se localiza em região urbana central.

The purpose of this paper was to quantify the copper and lead present in the water and sediments of the Aratimbó lake, located in the city of Umuarama, in the state of Paraná, and to analyze the abiotic indicators of temperature and pH of the water. The samples were taken in triplicate at three different points, on two different days - the first during dry weather, before the rain, and the other after the first rain. The temperature and pH of the samples were verified in locu and the metals were determined through Atomic Absorption Spectrophotometry by flame. The pH results obtained satisfactory values ​​at all points. Regarding the concentration of metals in the water, a higher concentration of lead was observed before the rain and a concentration below the limit was observed only for copper in point 1. In the sediment, the concentration of both metals was higher before the rain, except in point three, where the concentration was also higher after the rain. It is possible that the highest concentration values ​​for the metals were higher in the dry period due to the increased volume of water after rainfall, with higher dilution of the metals, as well as the attribution of contamination to the anthropogenic activities, since the lake is located in a central urban region.

Evaluación toxicológica y de metales (Cu, Pb, Ni, Zn y Cd) en el sedimento del reservorio Paiva Castro en Mairiporã-SP, Brasil / Toxicological and metal evaluation (Cu, Pb, Ni, Zn and Cd) in the sediment of Paiva Castro reservoir, in Mairiporã-SP, Brasil

El hombre ha cambiado el ambiente para sostener la demanda global de recursos naturales como el agua. La gestión de los cuerpos hídricos tiene que ser constante, con el propósito preventivo y correctivo, dependiendo del estado de antropización de cada sistema. El objetivo de este artículo fue analizar la toxicidad del sedimento y los metales Mn, Zn, Pb, Ni, Cd y Cu, en el reservorio Paiva Castro, que abastece la Region Metropolitana de São Paulo. Se realizaron 2 muestreos (Mayo 2011, estación seca y Enero de 2012, estación húmeda) y se analizaron 5 puntos próximos a la captación de agua por la Companhia de Saneamento Básico do Estado de São Paulo. Se realizaron ensayos de toxicidad aguda y crónica en sedimento, a través de ensayos biológicos con el cladócero Daphnia similis y el insecto Chironomus xanthus. El tratamiento de datos se realizó con el test de Fisher (mortalidad). El nivel de asociación entre las variables en sedimento y en los test ecotoxicológicos fueron evaluados por test no-paramétricos, a través del coeficiente de correlación de Spearman's. Los resultados del presente trabajo señalaron bajas concentracones de metales en el sedimento del área de estudio y ausencia de toxicidad en los organismos ensayados. Se puede concluir que área estudiada del reservorio Paiva Castro se encuentra poco impactada por los metales, sin efectos directos sobre la calidad de vida los organismos bentónicos: D. similis y C. xanthus.

Man had changed the natural environment in an attempt trying to supply the global demand for resources. The management of the hydric bodies has to be constant, with preventive and corrective purpose, depending on the eutrophization state of each one. The objective of this article was to analyze the sediment toxicity and the metals Mn, Zn, Pb, Ni, Cd and Cu, in the Paiva Castro reservoir, that supply the Metropolitan Region of São Paulo. It was made 2 collections (May 2011, dry season and January 2012, wet season). It was analyzed 5 points next to the water captation station by the Companhia de Saneamento Básico do Estado de São Paulo. The sediment was analyzed as for acute and chronic toxicity through bioassays with the cladocerans Daphnia similis and the insect Chironomus xanthus. Data treatment was done with Fisher Exact Test (mortality). The association level between the variables in sediment and ecotoxicological tests was available in non-parametric tests, through the Spearman's correlation coefficient. Oriented on the results presented in this work, pointing low concentrations of heavy metals in the sediments presented in the collect local, and the absence of toxicity, we can say that at this reservoir, at least in the collect area, it's low impacted, not implicating in direct interferences in the quality of life of benthonic organisms.

Growth of tropical tree species and absorption of copper in soil artificially contaminated / Crescimento de espécies arbóreas tropicais e absorção de cobre em solo artificialmente contaminado

Abstract Reclamation of copper contaminated sites using forest species may be an efficient alternative to reduce the negative impact. The aim of this study was to quantify the growth and evaluate the quality of seedlings of native species at different doses of copper in the soil. The experimental design was completely randomized, with seven replications in a factorial arrangement (3×9), using three indigenous species of plants (Anadenanthera macrocarpa, Mimosa scabrella and Apuleia leiocarpa) and nine doses of copper in the soil (0, 60, 120, 180, 240, 300, 360, 420 and 480 mg kg–1).The experiment was carried out in a greenhouse which the seedlings were grown for 180 days. The experimental units were plastic pots of 125 cm3 filled with Oxisol. The results indicated that the levels of copper applied to the soil decreased the quality of seedlings and growth of Apuleia leiocarpato a lesser extent compared with Mimosa scabrella and Anadenanthera macrocarpa. Anadenanthera macrocarpa was the forest species that resulted in the lowest copper translocation from roots to shoots. In addition, the Apuleia leiocarpa exhibited high resistance and tolerance for copper in the soil and also, it is highlighted an ability for copper phytoremediation.

Resumo A recuperação de áreas contaminadas com cobre utilizando espécies florestais pode ser uma alternativa eficiente para reduzir o impacto negativo deste elemento nestas áreas. O objetivo deste estudo foi quantificar o crescimento e avaliar a qualidade de mudas de espécies nativas em diferentes doses de cobre no solo. O delineamento experimental foi inteiramente casualizado, com sete repetições, em um esquema fatorial (3×9), utilizando-se três espécies nativas de plantas (Anadenanthera macrocarpa, Mimosa scabrella e Apuleia leiocarpa) e nove doses de cobre no solo (0, 60, 120, 180, 240, 300, 360, 420 e 480 mg kg–1). O experimento foi realizado em casa de vegetação, onde as plantas foram cultivadas por 180 dias. As unidades experimentais foram vasos de plástico de 125 cm3, preenchidos com Latossolo Vermelho distrófico. Os resultados indicaram que o nível de cobre aplicado ao solo reduziu a qualidade de plântulas e crescimento de Apuleia leiocarpa para um menor grau comparado com bracatinga e angico. Anadenanthera macrocarpa foi a espécie florestal que apresentou menor translocação de cobre a parte aérea das mudas. Além disso, a Apuleia leiocarpa exibiu elevada tolerância para o cobre no solo e também destaca-se mostrando uma capacidade para fitoremediação de áreas contaminadas com cobre.

Acute copper toxicity in juvenile fat snook Centropomus parallelus (Teleostei: Centropomidae) in sea water

Three experiments were designed to assess the accumulation and acute toxicity of copper (Cu) in juvenile fat snook Centropomus parallelus. The first experiment was performed to determine the 96-h lethal concentration (LC50) of Cu. The second experiment was designed to assess the effects of sublethal concentrations of Cu (0.47 and 0.94 mg/L), while the third one allowed us to test the recovery capacity of fish exposed to the sublethal concentrations Cu and kept in sea water without Cu addition. The LC50 value for Cu was found to be 1.88 mg/L Cu. Fish exposed to the sublethal concentrations of Cu showed a significant accumulation of Cu in gills at 96 h respect to the control ones (0.43 µg/g Cu). No significant difference was observed in the accumulation of Cu in gills between fish exposed to 0.47 mg/L (1.09 µg/g Cu) and 0.94 mg/L (1.26 µg/g Cu). Exposure (24 and 96 h) to the sublethal concentrations of Cu tested induced DNA damage in the erythrocytes. The results show that acute exposure to sublethal concentrations induces Cu accumulation and DNA damage in fish, these effects being recovered after 240 h in sea water without Cu addition.

Três experimentos foram realizados para avaliar o acúmulo e toxicidade aguda do cobre (Cu) em juvenis de robalo-peva Centropomus parallelus. O primeiro experimento foi realizado para determinar a concentração letal (96h-CL50) de Cu. O segundo experimento foi realizado para avaliar os efeitos de concentrações subletais de Cu (0,47 e 0,94 mg/L), enquanto o terceiro permitiu testar a capacidade de recuperação dos peixes expostos a concentrações subletais do Cu e posteriormente mantidos em água do mar sem acréscimo de Cu. O valor de LC50 encontrado para o Cu foi de 1,88 mg/L. Os peixes expostos as concentrações subletais de Cu mostraram um acúmulo significativo nas brânquias em relação ao controle em 96 h de exposição (0,43 µg/g Cu). Nenhuma diferença significativa foi observada entre os peixes expostos a 0,47 mg/L de Cu (1,09 µg/g) e 0,94 mg/L de Cu (1,26 µg/g). A exposição (24 e 96 h) para as concentrações subletais de Cu induziram danos no DNA. Os resultados mostram que a exposição aguda a concentrações subletais induz o acúmulo de Cu e danos ao DNA nas brânquias dos peixes, onde estes efeitos são recuperados após 240 h em água do mar sem adição de Cu.

Comparative acute toxicity of gallium(III), antimony(III), indium(III), cadmium(II), and copper(II) on freshwater swamp shrimp (Macrobrachium nipponense)

BACKGROUND: Acute toxicity testing were carried out the freshwater swamp shrimp, Macrobrachium nipponense, as the model animal for the semiconductor applied metals (gallium, antimony, indium, cadmium, and copper) to evaluate if the species is an suitable experimental animal of pollution in aquatic ecosystem. RESULTS: The static renewal test method of acute lethal concentrations determination was used, and water temperature was maintained at 24.0 ± 0.5°C. Data of individual metal obtained from acute toxicity tests were determined using probit analysis method. The median lethal concentration (96-h LC50) of gallium, antimony, indium, cadmium, and copper for M. nipponense were estimated as 2.7742, 1.9626, 6.8938, 0.0539, and 0.0313 mg/L, respectively. CONCLUSIONS: Comparing the toxicity tolerance of M. nipponense with other species which exposed to these metals, it is obviously that the M. nipponense is more sensitive than that of various other aquatic animals.

Copper resistance of different ectomycorrhizal fungi such as Pisolithus microcarpus, Pisolithus sp., Scleroderma sp. and Suillus sp.

Environments contaminated with heavy metals negatively impact the living organisms. Ectomy­corrhizal fungi have shown important role in these impacted sites. Thus, this study aimed to evaluate the copper-resistance of ectomycorrhizal fungi isolates Pisolithus microcarpus - UFSC-Pt116; Pisolithus sp. - UFSC-PT24, Suillus sp. - UFSM RA 2.8 and Scleroderma sp. - UFSC-Sc124 to different copper doses in solid and liquid media. The copper doses tested were: 0.00, 0.25, 0.5, 0.75, 1.0 and 1.25 mmol L-1 in the solid medium and 0.00, 0.32, 0.64 and 0.96 mmol L-1 in the liquid medium. Copper was amended as copper sulphate in order to supplement the culture medium MNM at pH 4.8, with seven replicates to each fungus-dose combination. The fungal isolates were incubated for 30 days at 28 °C. UFSC-Pt116 showed high copper-resistance such as accessed by CL50 determinations (concentration to reduce 50% of the growth) as while as UFSC-PT24 displayed copper-resistance mechanism at 0.50 mmol L-1 in solid medium. The UFSC-PT24 and UFSC-Sc124 isolates have increased copper-resistance in liquid medium. The higher production of extracellular pigment was detected in UFSC-Pt116 cultures. The UFSC-Pt116 and UFSC-PT24 isolates showed higher resistance for copper and produced higher mycelium biomass than the other isolates. In this way, the isolates UFSG-Pt116 and UFSC-PT24 can be important candidates to survive in copper-contaminated areas, and can show important role in plants symbiosis in these contaminated sites.

Evaluation of copper resistant bacteria from vineyard soils and mining waste for copper biosorption

Vineyard soils are frequently polluted with high concentrations of copper due application of copper sulfate in order to control fungal diseases. Bioremediation is an efficient process for the treatment of contaminated sites. Efficient copper sorption bacteria can be used for bioremoval of copper from contaminated sites. In this study, a total of 106 copper resistant bacteria were examined for resistance to copper toxicity and biosorption of copper. Eighty isolates (45 from vineyard Mollisol, 35 from Inceptisol) were obtained from EMBRAPA (Empresa Brasileira de Pesquisa Agropecuária) experimental station, Bento Gonçalves, RS, Brazil (29º09'53.92''S and 51º31'39.40''W) and 26 were obtained from copper mining waste from Caçapava do Sul, RS, Brazil (30º29'43.48''S and 53'32'37.87W). Based on resistance to copper toxicity and biosorption, 15 isolates were identified by 16S rRNA gene sequencing. Maximal copper resistance and biosorption at high copper concentration were observed with isolate N2 which removed 80 mg L-1 in 24 h. Contrarily isolate N11 (Bacillus pumilus) displayed the highest specific copper biosorption (121.82 mg/L/OD unit in 24 h). GenBank MEGABLAST analysis revealed that isolate N2 is 99 percent similar to Staphylococcus pasteuri. Results indicate that several of our isolates have potential use for bioremediation treatment of vineyards soils and mining waste contaminated with high copper concentration.

Estudio de niveles séricos de cobre y zinc y su posible rol como indicadores de dismenorrea primaria / Study of serum levels of copper and zinc and its possible roll like indicators of primary dysmenorrheal

En este trabajo se ha determinado el contenido de cobre y zinc en suero sanguíneo de 39 mujeres con edades comprendidas entre 17 y 21 años de edad, n = 25 con dismenorrea primaria diagnosticada clínicamente y n = 14 grupo control con diagnóstico negativo para la patología estudia-da. Las muestras de suero sanguíneo se obtuvieron antes (a) y durante (d) la menstruación. Fueron analizadas por Espectroscopia de Absorción Atómica con inyección en flujo continuo (EAA-IFC). Las concentraciones obtenidas fueron de 1,56 ± 0,43 mgL-1 de cobre y 1,28 ± 0,34 mgL-1 de zinc antes de la menstruación y 1,06 ± 0,23 mgL-1 de co bre y 1,21 ±0,4 mgL-1 de zinc durante la menstruación, para el grupo control y 1,64 ± 0,52 mgL-1 de cobre y 1,88 ± 0,61 mgL-1 de zinc antes de la menstruación y 1,21 ± 0,34 mgL-1 de cobre y 0,94 ± 0,27 mgL-1 de zinc durante la mens ruaciónt en el grupo estudio. Los elementos determinados en ambos grupos antes del periodo menstrual, no observaron diferencias significativas con p= 0,63 y 0,16 para cobre y zinc, respectivamente. Sin embargo, durante la menstruación, fue evidente una diferencia con p < 0,005. Este resultado nos hace presumir, que el aumento brusco de los niveles séricos de cobre se deba probablemente por la alteración de las prostaglandinas, siendo este bioelemento uno de los responsables de las molestias que se presentan en esta patología y otros efectos secundarios motivados por el descenso brusco del zinc. La relación cobre/zinc se calculo en ambos grupos, con valores de 1,47 y 1,06 antes y durante, respectivamente para el grupo control y para el grupo bajo estudio 1,36 y 1,97 antes y durante, respectivamente.

In this work has determined the copper content and zinc in sanguineous serum of women with primary dysmenorrheal diagnosed clinically and a group of women control with I diagnose negative for the studied pathology. The samples of sanguineous serum taken before (a) and during (d) the second day of the menstruation come from 39 women in ages between 17 and 21 years of age, n = 14 controls and n = 25 women with primary dysmenorrheal. They were processed by Spectroscopy of Atomic Absorption with injection in con-tinuous flow (EAA-IFC). The concentrations obtained of 1.56± 0.43 mgL-1 of copper and 1.28± 0.34 mgL-1 of zinc before menstruation and 1.06 ±0.23 mgL-1 of copper and 1.21 ±0.4 mgL-1 of zinc during the menstruation (second day), for the group control and 1.64 ±0.52 mgL-1 of copper and 1.88 ±0.61 mgL-1 of zinc before menstruation and 1.21± 0.34 mgL-1 of copper and 0.94± 0.27 mgL-1 of zinc during the menstruation in the group study. In relation to the serum levels it initiates to them in both groups were not significant differences (p = 0.63). The zinc, observe he himself behavior, between both groups before the menstruation was not observed significant differences with p = 0.16, nevertheless, during the menstruation I throw p = 0.0033. This result evi-dence that indeed the copper is affected by the alteration of the prostaglandins, being probably one of the people in charge of the annoyances that appear in this pa hologyt and other indirect effect motivated by the abrupt reduction of the zinc, such as alteration in the synthesis and excretion of the hormone stimulating follicle (FSH), of the luteinizante hormone (LH), abnormal development of the ovary, alterations of the menstrual cycle. The copper/zinc re ationl I calculate both groups, with values of 1.47 and 1,06 before and during res pectively for the group control and the group under study 1,36 and 1.97 before and during respectively.

Isolation and molecular characterization of some copper biosorped strains

TThe biosorption of copper [II] from aqueous solution using different bacterial strains was studied. Copper-biosorbing bacteria were isolated from tannery effluent collected from Borg Al-Arab, Alexandria, Egypt. These isolates displayed different degrees of copper biosorption under aerobic conditions. Based on 16S rDNA gene sequence analysis, three of them [S2, S5 and S7] were identified as Chryseobacterium sp., Enterobacter sp. and Stenotrophomonas sp., respectively. Initial copper [II] ion concentrations from 25-250 mg/L at constant temperature 30°C were studied. The residual copper [II] concentration and its toxicity effect in solution were determined using atomic absorption spectrophotometer and bioluminescent bioreporter. The bioluminescence inhibition of strain [S5] reached to 91.4% as compared with the strain [S7] reached to 83.3% at 225 mg/L of copper ion where the maximum biosorption efficiency for S5 and S7 were 71% and 70.1% correspondingly using atomic absorption. The biolumi-nescent bioreporter was proved to be fast and accurate technique for measurement the toxicity effect of residual copper [II] in solution

Cytogenetical and ultrastructural effects of copper on root meristem cells of Allium sativum L

Different copper concentrations, as well as different exposure times, were applied to investigate both cytogenetical and ultrastructural alterations in garlic (Allium sativum L.) meristem cells. Results showed that the mitotic index decreased progressively when either copper concentration or exposure time increased. C-mitosis, anaphase bridges, chromosome stickiness and broken nuclei were observed in the copper treated root tip cells. Some particulates containing the argyrophilic NOR-associated proteins were distributed in the nucleus of the root-tip cells and the amount of this particulate material progressively increased with increasing exposure time. Finally, the nucleolar material was extruded from the nucleus into the cytoplasm. Also, increased dictyosome vesicles in number, formation of cytoplasmic vesicles containing electron dense granules, altered mitochondrial shape, disruption of nuclear membranes, condensation of chromatin material, disintegration of organelles were observed. The mechanisms of detoxification and tolerance of copper are briefly discussed.

Impact of copper on the oxidative metabolism of the fry of common carp, Cyprinus carpio (Linn.) at different pH.

In order to evaluate the impact of copper on the energetics of a fish, the levels of glucose, glycogen, pyruvate and lactate, the rate of tissue oxygen consumption and the activities of glycogen phosphorylase, isocitrate dehydrogenase (ICDH), succinate dehydrogenase (SDH) and lactate dehydrogenase (LDH) were estimated in the whole body of the fry of Cyprinus carpio immediately after 1, 7, 15 and 30 days on exposure to a sublethal concentration of copper 0.08 mgl(-1) at pH 7.5 (normal), 6.0 (weak acidic) and 9.0 (weak alkaline). Aprogressive increase in glucose level and glycogen phosphorylase activity with the corresponding decrease in glycogen level over the time of exposure at pH 7.5 indicated glycogenolysis. Increase in the rate of oxygen consumption, pyruvate level and ICDH and SDH activities at days 1 and 7 (day 1 > 7) followed by their decrease at days 15 and 30 (day 15 < 30) at pH 7.5 indicated an initial elevation in the energetics of the fish fry with a gradual suppression of it on prolonged exposure. During this period the animal might have relied more on energetically less efficient glycolysis as evident by the progressive increase in the level of lactate and LDH activity. The degree of glycogenolysis was relatively more at pH 6.5 than at pH 7.5. At that pH, a progressive decrease in glucose level with an increase in the pyruvate and lactate levels and in LDH activity and a decrease in the rate of oxygen consumption and ICDH and SDH activities revealed greater reliance of the fish on anaerobic glycolysis than on oxidative metabolism. At pH 9.0 also the fish fry initially exhibited glycogenolysis, but gradually it came to normal on day 30 (day 1 > 7 > 15 > 30). Decrease in the glucose level, increase in pyruvate level, rate of oxygen consumption, and ICDH and SDH activities at all the days of exposure suggested an elevation in oxidative metabolism, but it also came to normal on prolonged exposure. Even the lactate level and LDH activity initially increased but gradually reached to normal on day 30. These results indicated that copper suppresses the energetics of the fish fry at pH 6.0, elevates at pH 9.0 relative to the changes at pH 7.5 suggesting that the toxicity of copper is dependent on pH of the water.

Impact of copper on haematological profile of freshwater fish, Channa punctatus.

The present study was to evaluate heavy metal toxicity stress symptoms in fish blood during long-term exposure of sublethal concentration of copper sulphate (CuSO4). The effects of copper on various heamatological parameters were evaluated exposing Indian fresh water fish, Channa punctatus to a sub-lethal concentration of copper sulphate (0.36 mg l(-1)) for different periods. Exposure of fish to copper showed a significant decrease in the haemoglobin (Hb) content from 10.73 to 6.60%, red blood cells (RBC) from 2.86 to 1.84 x 10(6)/mm3 and packed cell volume (PCV) from 31.00 to 23.33% at the end of 45th day as compared to control. Whereas the white blood cells (WBC) increased from 60.00 to 92.48 x10(3)/mm3, clotting time (CT) from 27.66 to 43.00 second, erythrocyte sedimentation rate from 5.0 to 13.66 mm/hr and mean corpuscular volume from 108.11 to 126.85 cmu, significantly, with increase in exposure periods. Although mean corpuscular haemoglobin concentration (MCHC) and mean corpuscular haemoglobin (MCH) values showed significant increase during 15 and 30 days exposures, both the values were found significantly increased at the end of the experimental period. The differential leucocytes count (DLC) showed a significant increase in the populations of lymphocytes and eosinophils, whereas neutrophils, monocytes and basophiles were found decreased after exposure to copper.

Effects of copper on chlorophyll, proline, protein and abscisic acid level of sunflower (Helianthus annuus L.) seedlings.

The effect of copperchloride (CuCl2) on the level of chlorophyll (a+b), proline, protein and abscisic acid in sunflower (Helianthus annuus L.) seedlings were investigated Control and copper treated (0.4, 0.5 and 0.6 mM) seedlings were grown for ten days in Hoagland solution. Abscisic acid content was determined in root, shoot and leaf tissues of seedlings by HPLC. Copper stress caused significant increase of the abscisic acid contents in roots, shoots and leaves of seedlings. The increase was dependent on the copper salt concentration. Enhanced accumulation of proline in the leaves of seedlings exposed to copper was determined, as well as a decrease of chlorophyll (a+b) and total protein (p < 0.05 or p < 0.01). It was observed that the level of chlorophyll (a+b) and total protein (p < 0.05 or p < 0.01) remarkably decreased as copper concentration increased to 0.6 mM, although the levels of proline and abscisic acid in the leaves of plants were increased--a dose-depended behavior The same trends were also observed with the level of abscisic acid of stems and roots. Copper has dose- depended effects on chlorophyll, proline, protein and abscisic acid level of sunflower (Helianthus annuus L.) seedlings. Thus, we assumed that copper levels increase above some critical points seedling growth get negative effects. This assumption is in line with previous findings.

Life table demography of Ceriodaphnia dubia (Cladocera) exposed to copper at different levels and periods.

We studied the demographic responses of Ceriodaphnia dubia exposed to copper at 3 different levels (0.1, 0.2 and 0.4 mg l(-1)) and at 4 exposure periods (3, 6, 12 and 24 hr) in addition to controls (without the heavy metal). The tested levels of Cu concentration and the chosen periods of exposure affected some demographic variables (average lifespan, gross and net reproductive rates and generation time). Depending on the heavy metal concentration and the period of exposure, the average lifespan of C. dubia varied from 18 to 24 day Net reproductive rates (NRR) ranged from 50 to 80 offspring per female per lifespan, while the rate of population increase (r) varied from 0.30 to 0.34 per day Though gross reproductive rates (especially at low and intermediate copper levels and shorter duration of exposure) showed significantly higher values than controls, both NRR and r were either unaffected or decreased. It is therefore necessary to regulate the release of untreated industrial effluents containing copper into freshwater ecosystems. Our study thus highlights the importance of including, not only toxicant concentrations but also exposure time, in ecotoxicological evaluations.

Response of wheat seed germination and seedling growth under copper stress.

The experiment was performed to study the seed germination and seedling growth in wheat under the influence of different concentrations of copper. The germination %, plumule and radicle length, and number of lateral roots decreased with increase in copper concentration (5, 25, 50 and 100 mgl(-1)). Total chlorophyll contents declined on 14th day from 1.605 of control to 1.581, 1.242, 1.275 and 1.107 mg g(-1) fresh weight in respective treatments. Similarly, on 21st day the decline in total chlorophyll contents was 1.288, 1.123, 1.077 and 0.985 mg g(-1) fresh weight in respective treatments against 1.724 of control. Likewise the pheophytin contents also declined showing the same pattern. However, carotenoid contents increased in different treatments, ranging between 0.366 to 0.464 mg g(-1) fresh weight in comparison to control (0.328) on 14th day, but showed adverse effects on 21st day as the carotenoid contents decreased in different copper treatments. The activity of amylase was found to be gradually reduced 14th day from 29.73 of control to 27.80, 27.33, 21.86 and 20.00 mg g(-1) and at 21st day from 14.40 of control to 11.46, 11.01, 9.86 and 5.60 mg g(-1) fresh weight with increase in concentrations of copper The catalase activity increased 14th day from 97.33 of control to 134.66, 161.33, 216.00 and 232.00 and on 21st day from 140.00 of control to 245.33, 274.66, 278.66 and 300.66 ml H2O2 hydrolyzed/g fresh weight in different increased concentrations of copper Similarly the peroxidase activity was also increased with increase in copper concentration. Likewise fresh weight and moisture contents decreased with increase in copper concentration. The dry weight was increased with increase in concentration of copper treatment. Total protein contents were initially decreased on 14th day from 80.69 of control to 66.75, 60.41, 56.41 and 48.48 microg/mg and on 21st day 81.37 of control to 67.06, 62.31, 54.92 and 46.47 microg/mg fresh weight in different copper concentrations respectively Sugar contents were significantly decreased in all the doses of copper on both 14th and 21st days i.e. (5.53 of control to 4.76, 3.69, 3.68 and 2.86 microg/mg in different copper treatments on 14th day and 4.81 of control to 4.49, 3.40, 2.79 and 2.15 microg/mg on 21st day respectively.

Copper toxicosis in sheep fed dairy cattle ration in São Paulo, Brazil

Intoxicação crônica por cobre foi observada em um rebanho de 20 ovinos no município de Franca, São Paulo. Três meses após o início do arraçoamento com feno e ração concentrada peletizada para bovinos de leite, seis animais apresentaram anorexia, icterícia severa e urina marrom escura, e vieram a óbito. Diagnosticou-se doença hemolítica com base em sinais clínicos, alterações macroscópicas observadas na necropsia e observações histológicas. A necropsia todos os ovinos apresentaram icterícia severa, fígado amarelado com padrão lobular evidente e rins escuros. As principais alterações histológicas incluíram necrose hepática periacinar e nefrose hemoglobinúrica. Acúmulos de cobre foram demonstrados nos hepatócitos e macrófagos pela coloração rodamina e níveis elevados de cobre mediante espectrofotometria de absorção atômica no soro, fígado e rins de dois ovinos afetados e na ração fornecida.

Intoxicação cúprica acumulativa experimental em bovinos / Induced accumulative copper poisoning in catle

O presente trabalho objetivou analisar variáveis clínicas, sangüíneas eos teores de cobre hepático de bovinos submetidos à intoxicação cúprica acumulativa (ICA), por meio do fornecimento de quantidades crescentes de cobre, Com tal objetivo, foram utilizados 10 bovinos(mestiços) jovens, aleatoriamente distribuídos em seis animais no grupo suplementado com cobre (BOV Cu) e quatro animais no grupo controle (BOV). Diariamente, o grupo BOV Cu recebeu por meio de cânula ruminal 2 mg Cu /kg/PV (CuSO4.5H2O) sendo esta dose acrescida de mais 2 mg/kg/PV a cada semana, até o término do experimento (105 dias). Foram realizadas três biópsias hepáticas (Dia0 - dia 45 - dia 105) em todos os animais para determinação da concentração de Cu e Zn neste órgão. Quinzenalmente, foi realizado exame clínico, pesagem dos animais e coleta de amostras de sangue. Três bovinos do grupo BOV Cu manifestaram quadro laboratorial e/ou clínico sugestivo de intoxicação cúprica acumulativa (ICA), vindo a sucumbir em seguida. Destaca-se a presença de dois quadros clínicos diferentes, o clássico (n = 1) e um atípico (n = 2), caracterizado pelo destacado acúmulo de cobre hepático, hiporexia progressiva seguida de anorexia, desidratação, redução dos movimentos de ruminais, oligúria, acentuada apatia e morte, mas sem apresentar hemoglobinúria. Parte dos bovinos se mostraram resistentes à ICA a despeito da administração de altas quantidades cobre. Bovinos com ICA aumentaram a concentração de zinco hepático nas fases finais da intoxicação. Nenhum animal apresentou quadro de insuficiência renal.

The principal objective of this project was to evaluate clinical and hematological alterations and the hepatic concentration of Cu in cattle with accumulative copper poisoning, by infusion of increasing doses of copper. Ten cattle yearling steers were randomly distributed into acopper supplemented group (BOV Cu; n = 6) and one control group (BOV; n = 4). The group BOV Cu received initially 2 mg Cu/kg/BW(as CuSO4.5H2O) daily for one week; every following week, up to the end of the experiment (105th day), this initial dose was increased by 2mg Cu/kg/BW. Three liver biopsies were realized during the experiment (day zero, 45th, and 105th day) to evaluate the degree of copper accumulation in this organ; the body weight and clinical examination was monitored every 15 days, when blood samples were taken. Three cattle supplemented with copper demonstrated clinical manifestations and/or laboratory findings consistent with accumulative copper poisoning (ACP), and died. Two distinct clinical manifestations were observed, one classical (n = 1) and another atypical(n = 2) characterized by remarkable high levels of liver copper, progressive hyporexia followed by anorexia, dehydration, severe apathy, decreased rumen movements, oliguria, and death. Some animals were resistant to ACP despite high accumulative levels of hepatic copper. Cattle with ACP increased their zinc liver concentration at the final stages of the poisoning. No poisoned animals developed renal insufficiency.