ABSTRACT
Understanding the effects of heavy metals in aquatic ecosystems is of significant importance due to their potential to bioaccumulate at various trophic levels and induce damage in DNA. Mercury is considered one of the most dangerous heavy metals, causing chromosomal breakage (clastogenic event) or spindle dysfunction (aneugenic event), that can lead to the formation of encapsulated chromatin into a separate smaller nucleus, generally referred to as a micronucleus. We evaluated the sensitivity of the micronucleus test in the neotropical cichlid Andinoacara rivulatus (Günther 1860). The fish were divided into four groups of 16 individuals, and each group was placed in separate aquaria (140 L) provided with filtered water and constant aeration. Fish were exposed to mercury chloride (HgCl2) at doses 0.1, 0.25, and 0.50 mg/kg body weight, administered by intraperitoneal (IP) injection. Fish from the control group were injected with a physiologic solution. The following erythrocyte anomalies were identified: erythrocytes with micronuclei varying to some extent in size and position in the cytoplasm, blebbed nucleus, binucleated cell, nuclei showing a deep invagination toward the center (notched nuclei). Examination of blood smears demonstrated a higher level of micronucleus and notched erythrocytes in fish injected with HgCl2 than in the controls. There were significant differences in the frequency of micronucleated and notched erythrocytes among the groups exposed to mercury. Linear regression analysis revealed a positive relationship between the frequency of micronucleated and notched erythrocytes (P< 0.0001), with a moderately strong correlation coefficient (R= 0.737). We propose that, in addition to the two so far known mechanisms of micronucleus formation (spindle apparatus damage and chromosomal ruptures), chromatin fragmentation in notched nuclei resulting from a combination of the cytotoxic effects of mercury and mechanical stress, may be a third mechanism of micronuclei genesis.
Comprender los efectos de los metales pesados en los ecosistemas acuáticos es de gran importancia debido a su potencial de bioacumulación en los diversos niveles tróficos y de inducir daños en el ADN. El mercurio, considerado uno de los metales pesados más peligrosos, causa fracturas cromosómicas (evento clastogénico) o disfunción del huso mitótico (evento aneugénico), y puede conducir a la formación de fragmentos de cromatina encapsulada en un núcleo separado más pequeño, generalmente denominado micronúcleo. En este trabajo se evalúa la sensibilidad del test de micronúcleos en el cíclido neotropical Andinoacara rivulatus (Günther 1860). Los peces fueron divididos en cuatro grupos de 16 individuos, y cada grupo se colocó en acuarios separados (140 L) provistos de agua filtrada y aireación constante. Los peces fueron expuestos al cloruro de mercurio (HgCl2) en dosis de 0.1, 0.25 y 0.50 mg/kg de peso corporal, administrada por inyección intraperitoneal (IP). Los peces del grupo control fueron inyectados con solución fisiológica. Se identificaron las siguientes anomalías en los eritrocitos: micronúcleos que varían en tamaño y posición en el citoplasma, núcleo con evaginaciones, células binucleadas núcleos con muesca (núcleos con una profunda invaginación hacia el centro). El examen de los frotis de sangre demostró un mayor nivel de eritrocitos micronucleados y con muesca en peces inyectados con HgCl2 que en los controles. Fueron detectadas diferencias significativas en la frecuencia de eritrocitos micronucleados y células con núcleos con muescas entre los grupos expuestos al mercurio. El análisis de regresión lineal reveló una relación positiva entre la frecuencia de eritrocitos micronucleados y con muescas (P< 0.0001), con un coeficiente de correlación moderadamente fuerte (R= 0.737). Se propone que, además de los dos mecanismos hasta ahora conocidos de formación de micronúcleos (daño del huso mitótico y rupturas cromosómicas), la fragmentación de la cromatina en núcleos con muesca probablemente a causa de la combinación de los efectos citotóxicos del mercurio y el estrés mecánico, podría ser un tercer mecanismo de génesis de micronúcleos.
ABSTRACT
This study aimed to determine the variables leukocytary, erythrocytary, frequency of micronuclei and nuclear abnormalities in peripheral erythrocytes of Prochilodus argenteus, Pimelodus maculatus and Myleus micans from São Francisco river basin, Minas Gerais State, during the summer and winter. Thrombocytes, hematocrit and leukocyte of P.argenteus series, was influenced by climatic conditions. In P. maculatus, not were significant differences for all leukocytes and thrombocytes. In M. micans, values are unprecedented in the literature, with MCV and absolute number of leukocytes found superior to other species of the family Characidae. The frequency of micronuclei and nuclear abnormalities in erythrocytes, the three species not showed seasonal differences between the sites. However, the highest values were found in summer. Furthermore, we observed a positive correlation between the increase in the percentage of micronuclei with nuclear abnormalities
O presente estudo teve como objetivo, determinar as variáveis leucocitárias, eritrocíticas, freqüência de micronúcleos e anomalias nucleares nos eritrócitos do sangue periférico de Prochilodus argenteus, Pimelodus maculatus e Myleus micans de um trecho da bacia do rio São Francisco, Minas Gerais, no verão e inverno. Trombóctios, hematócrito e a serie leucocítária de P.argenteus, foi influenciado pela sazonalidade. Em P. maculatus, não ocorreram diferenças significativas para todos os leucócitos e trombócitos. Em M. micans, os valores são inéditos na literatura, apresentando VCM e número absoluto de leucócitos superior ao encontrados para outras espécies da família Characidae. A freqüência de micronúcleos e de anomalias nucleares nos eritrócitos, nas três espécies não apresentou diferenças sazonais e entre os pontos amostrais. No entanto, os valores mais altos foram encontrados no verão. Além disso, foi possível observar correlação positiva entre o aumento da porcentagem de anomalias nucleares com micronúcleos.