Efecto Ecotoxicológico de la Mezcla de Aroclor 1254 y Plomo en el Bioindicador Daphnia magna / Ecotoxicological effect of the mixture of Aroclor 1254 and lead in the bioindicator Daphnia magna / Efeito ecotoxicológico da mistura de aroclor 1254 e chumbo no bioindicador Daphnia magna

Resumen Introducción: el bioensayo con la especie bioindicadora Daphnia magna Strauss, 1820 posibilita obtener el efecto ecotoxicológico agudo individual y en mezcla de diversos tóxicos químicos mediante la concentración letal media (CL50). Objetivo: determinar el efecto ecotoxicológico de la mezcla de aroclor 1254 y del plomo (Pb2+) sobre D. magna. Materiales y Métodos: se midió la CL50 siendo aroclor 1254 > Pb2+, y ulteriormente la concentración sin efecto ecológico (PNEC) para el efecto individual y en mezcla. Se contrastaron los valores de PNEC con la categoría 4 - E1 (lagunas y lagos) y E2 (ríos) del Estándar de Calidad Ambiental Peruano (Decreto Supremo N° 004 2017-MINAM -Perú). Resultados: los valores de CL50 disminuyeron de forma gradual a 48 h de exposición para ambos compuestos en forma individual y en mezcla. Se calculó la proporción - sinergia (PS) en relaciones de 1:1, 3:2 y 4:1 (aroclor 1254: Pb2+), obteniéndose mayores valores de PS en 3:2 y 4:1 en contrastación a 1:1. Conclusiones: la concentración de aroclor 1254 estimada de PNEC es similar al valor de la conservación del ambiente acuático categoría 4 - E1 y E2, y la concentración máxima para Pb2+ no es adecuada y debe reducirse para fortalecer la conservación de la biota acuática. Finalmente, el bioindicador D. magna determinó idóneamente la mezcla de aroclor 1254 y Pb2+.

Abstract Introduction: the bioassay with the bioindicator species Daphnia magna Strauss, 1820 makes it possible to obtain the individual acute ecotoxicological effect and in a mixture of various chemical toxics by means of the mean lethal concentration (LC50). Objective: to determine the ecotoxicological effect of the mixture of aroclor 1254 and lead (Pb2+) on D. magna. Materials and Methods: LC50 was measured with arochlor 1254> Pb2+, and subsequently the concentration without ecological effect (PNEC) for the individual effect and as a mixture. PNEC values were contrasted with category 4-E1 (lagoons and lakes) and E2 (rivers) of the Peruvian Environmental Quality Standard (Supreme Decree No. 004 2017-MINAM-Peru). Results: LC50 values gradually decreased after 48 h of exposure for both compounds individually and as a mixture. The proportion-synergy (PS) was calculated in ratios of 1:1, 3:2 and 4:1 (arochlor 1254: Pb2+), obtaining higher PS values in 3:2 and 4:1 in contrast to 1:1. Conclusions: the PNEC estimated arochlor 1254 concentration is similar to the conservation value of the aquatic environment category 4-E1 and E2, and the maximum concentration for Pb2+ is not adequate and should be reduced to strengthen the conservation of aquatic biota. Finally, the bioindicator D. magna ideally determined the mixture of aroclor 1254 and Pb2+.

Resumo Introdução: o bioensaio com as espécies bioindicadoras Daphnia magna Strauss, 1820 permite obter o efeito ecotoxicológico agudo individual e numa mistura de várias toxinas químicas por meio da concentração letal média (CL50). Objetivo: determinar o efeito ecotoxicológico da mistura de arocloro 1254 e chumbo (Pb2+) sobre D. magna. Materiais e Métodos: a CL50 foi medida com arocloro 1254>Pb2+, e posteriormente a concentração sem efeito ecológico (PNEC) para o efeito individual e como mistura. Os valores do PNEC foram contrastados com a categoria 4-E1 (lagoas e lagos) e E2 (rios) do Padrão de Qualidade Ambiental Peruano (Decreto Supremo n° 004 2017-MINAM-Peru). Resultados: Os valores de LC50 diminuíram gradualmente após 48 h de exposição para ambos os compostos individualmente e como uma mistura. A proporção-sinergia (PS) foi calculada em proporções de 1:1, 3:2 e 4:1 (arocloro 1254: Pb2+), obtendo-se valores de PS mais altos em 3:2 e 4:1 em contraste com 1:1. Conclusões: a concentração estimada de arocloro 1254 do PNEC é semelhante ao valor de conservação do ambiente aquático categoria 4-E1 e E2, e a concentração máxima de Pb2+ não é adequada e deve ser reduzida para fortalecer a conservação da biota aquática. Finalmente, o bioindicador D. magna determinou idealmente a mistura de arocloro 1254 e Pb2+.

Low dose and subchronic Aroclor 1254 induced histopathological changes in mice kidney

Present communication reports the low dose and subchronic duration-dependent histopathological changes afterexposure to Aroclor 1254 in the kidney tissue of Swiss albino mice. Polychlorinated biphenyls (PCBs) like Aroclor1254 congeners accumulate in tissues rich in lipids and remain inside for a long time, affecting the functionality of thecells in direct and indirect ways. The most commonly observed effects were skin ailments such as chloracne, rashes,and effects on kidney functions. Animal studies indicated that PCBs could affect the functionality of the kidney,thyroid, immune, and endocrine systems. Separate groups of mice were subjected to a daily oral dose of 0.1 mg/kgbody weight (BW)/day and 1 mg/kg BW/day Aroclor 1254, dissolved in corn oil, for four subacute exposure durations(7, 14, 21, and 28 days). Control groups were received only the corn oil (vehicle). Results revealed mainly exposureduration-dependent histopathological lesions such as dilation of the tubular cells, fragmentation of cytoplasm andloss of nuclear materials, formation of large vacuoles inside the cells, and necrosis even at very low doses of Aroclorintoxication. The present study reports predominantly exposure duration-dependent histopathological effects ofAroclor 1254 in the kidney tissue of mice. The study suggested that the subacute exposure to low doses of Aroclor1254 could cause significant irreparable structural deformities in the renal cells of the kidney tissue. Results alsoshowed that renal tubular cells were more affected showing severe necrosis

Avaliações toxicológicas das alterações genotóxicas e epigenóticas induzidas por Aroclor 1254 em testí­culo, espermatozóides, fí­gado e rim de camundongos / Toxicological assessments of genotoxic and epigenetic changes induced by Aroclor 1254 in testis, sperm, liver and kidney of mice

As bifenilas policloradas (PCBs) são um grupo de compostos hidrocarbonetos halogenados aromáticos, bioacumulativos em organismos vivos e persistente no ambiente. Além da atividade disruptora endócrina, os PCBs podem aumentar os níveis de espécies reativas de oxigênio (ROS), levando ao estresse oxidativo e alteração da metilação de DNA que são fatores importantes nas etiologias da hepatotoxicidade, infertilidade masculina e doença renal. Estes agentes tóxicos podem causar disfunção mitocondrial e distúrbios que afetam a produção de ATP, ROS e morte celular, ocasionando danos à saúde humana. O presente trabalho tem como objetivo investigar possíveis alterações genotóxicas e epigenéticas causadas pelo aroclor 1254 em fígado, rim e testículo, além de verificar a indução de estresse oxidativo e disrupção dos metabólitos intermediários do ciclo de Krebs nos referidos tecidos. Camundongos machos C57/BL6 foram expostos ao Aroclor 1254 em diferentes doses (5, 50, 500 e 1000 ug/kg) por gavagem, uma vez a cada três dias, durante 50 dias. Após a exposição, os animais foram eutanasiados, os órgãos coletados e espermatozoides obtidos a partir dos epidídimos. A peroxidação lipídica em plasma e tecidos foi avaliada pela quantificação de malonaldeído (MDA) por HPLC/DAD. Os níveis de intermediários da via glicolítica, do ciclo de Krebs, de alguns nucleotídeos e aminoácidos, marcas epigenéticas (5-mC e 5-hmC) e adutos de DNA (8-oxodG e CEdG) foram quantificados por HPLC-ESI-MS/MS. A abordagem de benchmark dose (BMD) foi utilizada para a modelagem dose resposta. Após exposição, não foram observadas diferenças significativas da variação da massa corporal, e a razão do peso testicular, fígado e rim por massa corporal. No tecido hepático, foi observado aumento da peroxidação lipídica. Houve redução significativa dos níveis de ATP, ADP, razão NADP+/NADPH, piruvato, malato, fumarato e glutamato. Observou-se redução significativa dos níveis de 5-mC e 5-hmC no DNA nuclear (nDNA), enquanto não foram observadas alterações dos níveis dos adutos. Em DNA mitocondrial (mtDNA) não foram observadas alterações nas marcas epigenéticas, no entanto foi obtido aumento significativo no aduto 8-oxodG após exposição ao Aroclor 1254. No tecido renal foi observado aumento significativo de MDA. Houve aumento significativo dos níveis de lactato e malato e reduções de ATP, ADP, glutamina, NAD+. Foi observada a hipohidroximetilação do mtDNA. As marcas 5-mC de mtDNA, 5mC de nDNA e adutos de DNA nuclear e mitocondrial não apresentaram diferenças após exposição a PCBs. Nos testículos foi verificada redução significativa dos níveis de glutamato, malato, succinato, fumarato e razão NADH/NAD+, hipohidroximetilção em mtDNA e hipermetilação em nDNA. Não foram observadas alterações de 5-mC em mtDNA e 5hmC em nDNA. Não foram verificadas alterações dos níveis de MDA e adutos em nDNA. Adicionando, foi observada redução dos níveis de 5-mC em DNA global de espermatozoide. Os limites inferiores do intervalo de confiança da BMD foram estimados para que estes marcadores possam ser usados na avaliação de riscos de PCBs. Os dados obtidos apontam o Aroclor 1254 como indutor de alterações do metabolismo intermediário, das marcas epigenéticas e estresse oxidativo. Essas alterações podem afetar vias celulares, levando à morte ou transformação, e aumentando o risco de doenças

Polychlorinated biphenyls (PCBs) are a group of aromatic halogenated hydrocarbon compounds, which bioaccumulate in living organisms and is persistent in the environment. Besides their endocrine disrupting activity, PCBs may increase the levels of reactive oxygen species (ROS), leading to oxidative stress and alter DNA methylation that are important factors in the etiology of liver toxicity, male infertility, and kidney disease. These toxic agents can cause mitochondrial dysfunction and disorders that affect the production of ATP, ROS and cell death, thereby leading to health-related problems. The present work aimed at investigating possible genotoxic and epigenetic changes caused by aroclor 1254 in the liver, kidney and testis, as well as determine the induction of oxidative stress and disruption of intermediate metabolites in these tissues. Male C57/BL6 mice were exposed to Aroclor 1254 at different doses (5, 50, 500 and 1000 µg/kg) by gavage, once every three days, for 50 days. After the exposure period, the animals were euthanized, organs collected, and sperms obtained from the epididymis. Lipid peroxidation in plasma and tissues was determined by quantification of malonaldehyde (MDA) using HPLC/DAD. The levels of intermediate metabolites, epigenetic marks (5-mC and 5-hmC) and DNA adducts (8-oxodG and CEdG) were quantified by HPLC-ESI-MS/MS. The Benchmark dose approach (BMD) was used for dose response modeling. No significant differences in body weight variation, testicular, liver and kidney weight to body weight ratio were observed after exposure. However, in hepatic tissues, an increase in lipid peroxidation was observed. There were significant decreases in the intermediate metabolites including the levels of ATP, ADP, pyruvate, NADP+/NADPH ratio, malate and fumarate, as well as glutamate. Significant reduction of 5-mC and 5-hmC levels in nuclear DNA (nDNA) were observed, whereas no changes were observed in DNA adducts. The epigenetic marks in mitochondrial DNA (mtDNA) were not changed; however, a significant increase was observed in 8-oxodG adduct after exposure to Aroclor 1254. In renal tissues, data showed a significant increase in MDA, while for the intermediate metabolites, the levels of lactate and malate were significantly elevated, whereas significant reductions were recorded for ATP, ADP, glutamine, and NAD+. Hypohydroxymethylation was observed in mtDNA. The 5-mC of mtDNA, 5mC of nDNA and nuclear and mtDNA adducts did not show differences after PCBs exposure. For the testicles, significant reductions in the levels of glutamate, malate, succinate, fumarate and NADH/NAD+ ratio were observed. The PCBs also induced hypohydroxymethylation in mtDNA and hypermethylation in nDNA, but there were no changes of 5-mC in mtDNA and 5-hmC in nDNA. A reduction of nDNA adducts 8-oxodG was observed. No changes were observed in the level of MDA and DNA adducts of nDNA. However, after PCBs exposure there was a significant decrease of 5-mC in global DNA of spermatozoa. The lower bound confidence interval on BMD, which were estimated for these markers can be used in the risk assessment of PCBs. Collectively, the data obtained in this study indicate that Aroclor 1254 induces alteration of intermediate metabolites, epigenetic marks and oxidative stress. These changes can adversely affect cells and cellular pathways, therefore increase the risk of cell death or transformation

Toxic effects of polychlorinated biphenyls (Aroclor 1254) on human sperm motility / 亚洲男科学杂志(英文版)

Polychlorinated biphenyls (PCBs) are common environmental contaminants that represent a considerable risk to reproductive toxicity in exposed human populations. Although some experimental studies have suggested an association between the levels of PCBs and semen quality, the direct effects of PCBs on human sperm parameters remain largely unexplored. To this aim, a short-term in vitro incubation experiment that better imitated the putative exposure of sperm to Aroclor 1254 (a commercial PCB mixture) in male reproduction tissue was conducted. Human sperm were incubated with various concentrations (0, 1, 5, or 25 mg l-1) of Aroclor 1254 for different amounts of time (3 and 6 h) in vitro. Sperm motility parameters were analyzed with computer-assisted sperm analysis (CASA). The proportion of sperm with high mitochondrial membrane potential (ΔΨm) and the levels of intracellular reactive oxygen species (ROS) were detected to explore the probable cause of sperm impairment. Human sperm exposed to continuous Aroclor 1254 exhibited: (i) reduced sperm motility and kinematic parameters, (ii) a proportion of sperm with high ΔΨm that decreased in a dose-dependent manner (P < 0.05), and (iii) increased levels of ROS compared with controls (P < 0.05). In conclusion, Aroclor 1254 can decrease sperm motility, which may culminate in increased ROS and general mitochondrial dysfunction, thus affecting the fertilization potential of sperm. Our findings suggest a broader understanding of the effect of Aroclor 1254 on human sperm.

Effect of gestational and lactational exposure to the polychlorinated biphenyl mixture aroclor 1254 on learning and memory and hippocampus in rat offspring / 中华行为医学与脑科学杂志

Objective To explore the effect of polychlorinated biphenyls (PCBs) on the learning and memory of gestational and lactational exposure to polychlorinated biphenyls in rats offspring.Methods The PCB mixture (A1254,0,5,10,20 mg/kg body weight) was administered to pregnant wistar rats every 3 days by gavage from gestational day (GD) 5 to postnatal day (PND) 20.To assess the effects on offspring following such exposure,Morris water maze test was performed to assess the learning and memory ability.Calcium concentration was assayed in hippocampus and the ultramicro structure was observed.Results After training for four days,the escaping latency in every group decreased significantly compared with the first day,especially the control group,the results in the fourth day decreased significantly with the early three days(P＜0.05).The offsprings of the 10 and 20 mg/kg had prolonged time of passing the aim target ((5.23± 1.16) s,(7.90±3.21) s,(11.74±6.56) s and (20.83± 8.38) s ; P＜0.05),decreased number of crossing platform ((4.14± 1.21),(3.00± 1.32),(2.65± 1.13),(2.42± 1.31) ; P＜0.05) and swimming time in the target area ((40.14±7.14)s,(33.76±5.58)s,(32.45±6.00)s and (30.63±5.10) s; P＜0.05)compared with those of the rats of control and 5 mg/kg groups.The calcium concentration increased ((121.16± 12.23) nM,(141.27±24.66) nM,(163.32±29.75) nM,(261.46±27.79) nM) and the ultramicro structure in hippocampus changed obviously in the high exposure group.Conclusion Gestational and lactational exposure to aroclor 1254 in rats could affect the leaming and memory ability of offsprings.

New Evidences of Neurotoxicity of Aroclor 1254 in Mice Brain: Potential of Coenzyme Q10 in Abating the Detrimental Outcomes

OBJECTIVES: The present subacute study was designed to evaluate the effect of coenzyme Q 10 (CoQ10) in the 28 days aroclor 1254 exposure induced oxidative stress in mice brain. METHODS: Biochemical estimations of brain lipid peroxidation (LPO), reduced glutathione (GSH), and activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and acetyl cholinesterase (AChE), and histopathological investigations of brain tissue were carried out. RESULTS: Oral exposure of aroclor 1254 (5 mg/kg) led to significant decrease in levels of GSH, and activities of SOD, CAT, GPx, and AChE, and increase in LPO. These aberrations were restored by CoQ10 (10 mg/kg, intraperitoneal injection [IP]). This protection offered was comparable to that of L-deprenyl (1 mg/kg, IP) which served as a reference standard. CONCLUSIONS: Aroclor 1254 exposure hampers the activities of various antioxidant enzymes and induces oxidative stress in the brains of Swiss albino mice. Supplementation of CoQ10 abrogates these deleterious effects of aroclor 1254. CoQ10 also apparently enhanced acetyl cholinesterase activity which reflects its influence on the cholinergic system.

Effects of low concentrations of a polychlorinated biphenyl, Aroclor 1254 on membrane bound ion dependent ATPases in mice liver.

Aroclor 1254, a polychlorinated biphenyl, is present in the environment in low concentration but references on its toxic effects on liver cell membrane proteins and the mechanism of actions are not abundantly available. Therefore, the present study was undertaken to investigate the low level, sub-acute dose and exposure duration dependent effects of Aroclor 1254 on total, Na+, K+, Ca2+ and Mg2+-ATPases of the mouse liver. The hypotheses tested in the present study were, (a) whether the low, environmentally available dose and the exposure durations of Aroclor 1254 affects the membrane-bound ion dependent ATPases, and (b) if a response was observed, whether it is a direct or indirect effects of the toxicant. Groups of mice were exposed to different doses (0.1 and 1mg kg-1 body weight d-1) and exposure durations (4 d, 8 d and 12 d) of Aroclor 1254. The results indicated significant exposure duration dependent changes in the specific activity of the selected membrane bound ATPases. As the observed changes were mostly enzyme stimulation after toxication through oral administration, the effects of the Aroclor were possibly indirect, through complex chain of reactions.