Marine shrimps as biomonitors of the Fundão (Brazil) mine dam disaster: A multi-biomarker approach.

The disruption of the Fundão dam released 43 million m3 of mine tailings into the Doce River until it flowed into the ocean through the estuary. The mine tailing changed the composition of metals in water and sediment, creating a challenging scenario for the local biota. We used multivariate analyzes and the integrated biomarker response index (IBR) to assess the impact of mine tailings on the bioaccumulation profile (As, Cd, Cr, Cu, Fe, Mn, Pb and Zn) as well as the biomarkers response in gills, hepatopancreas and muscle of shrimps sampled from different sectors during two dry seasons (dry1 and dry2) (Sep/Oct 2018; 2019) and two wet seasons (wet1 and wet2) (Jan/feb 2019; 2020). There was seasonal and local effect under bioaccumulation and biomarker response revealing that the pattern responses seen in each sector sampled changed according to the season. The greater IBR added to the strong association among the most metals tissue content (Cd, Cr, Cu and Mn) and sectors sampled during dry 1 suggests greater bioavailability of these metals to the environment in this period. Estuarine sectors stand out for high Fe bioavailability, especially during wet1, which seems to be associated with greater metallothionein content in hepatopancreas of shrimps. Native species of marine shrimps proved to be successful indicators of sediment quality besides being sensitive to water contamination by metals. The multi-biomarkers approach added to multivariate analysis supports the temporal and seasonal effects, signalizing the importance of continuous monitoring of the estuarine region to better know about the bioavailability of these metals, mainly Fe, and their long-term effects on the local biota.

Biochemical response and metals bioaccumulation in planktonic communities from marine areas impacted by the Fundão mine dam rupture (southeast Brazil).

The rupture of the Fundão dam (Mariana, MG, southeast Brazil) released a huge flood of mine tailings to Doce river basin and its adjacent coastal area, in November 2015. This catastrophic event exposed aquatic communities to metal contamination related to mine tailings, but its biological effects are still poorly understood. This study investigates how biochemical response related to metal exposure vary between locations and seasons during the years of 2018-2020, in planktonic communities (micro and mesoplankton). Marine microplankton collected in sectors in front and south of the Doce river mouth presented the highest lipid peroxidation (LPO) and induction of metallothioneins (MT). Mesoplankton collected in sectors in front and north of the Doce river mouth presented highest LPO, while MT in this size class did not respond to a clear spatial pattern. Our results showed that metals affected biomarkers in a non-linear pattern and highlighted the complex relationship between metals, biochemical parameters, and seasonality. The variation in biochemical biomarkers indicates physiological stress related to metals, once sectors contaminated by metals, especially Fe, Mn and Cd, presented stronger biochemical responses. Comparison of metal levels with bioaccumulation data collected before the impact indicates Fe, Cd, Cr and Cu more than 2-fold higher after disaster in sectors closer to the river. Literature showed that these sectors present zooplanktonic assemblages with lower biomass and biodiversity, suggesting that the opportunistic species that thrives in the area are also under biochemical stress, but possibly relies on repair or defense mechanisms. The physiological stress detected by this study is possibly related to the mine tailings, considering the metals that stood out and the proximity with the Doce river mouth. This suggests that the impacts related to the failure of Fundão dam are still affecting the marine planktonic community even three to four years after the environmental disaster.

High level of GHR nuclear translocation in skeletal muscle of a hyperplasic transgenic zebrafish.

It has been reported that nuclear translocation of growth hormone receptor (GHR) may directly activate cell proliferation in mammals and birds. However, this phenomenon has not yet been described in fish. Recently, we have developed a transgenic zebrafish that overexpresses GHR in a muscle-specific manner. Considering that this transgenic model exhibits hyperplasic muscle growth, the present work aims at verifying the relationship between GHR nuclear translocation and muscle cell proliferation. This relationship was evaluated by the phosphorylation state of the proliferative MEK/ERK pathway, expression of nuclear import-related genes, immunostaining of phospho-histone H3 (PH3) as a proliferation marker, and nuclear GHR localization. The results showed a significant decrease in the phosphorylation state of ERK1/2 proteins in transgenics. Moreover, there was an increase in expression of three out of four importin genes analyzed parallel to a large flow of GHR displacement toward and into the nucleus of transgenic muscle cells. Also, transgenics presented a marked increase in PH3 staining, which indicates cell proliferation. These findings, as far as we know, are the first report suggesting a proliferative action of GHR in fish as a consequence of its increased nuclear translocation. Thus, it appears that the nuclear migration of cytokine receptors is a common event among different taxonomic groups. In addition, the results presented here highlight the possibility that these membrane proteins may be involved more directly than previously thought in the control of genes related to cell growth and proliferation.

Relationships between multidrug resistance (MDR) and stem cell markers in human chronic myeloid leukemia cell lines.

The K562 cell line (chronic myeloid leukemia), sensitive to chemotherapy (non-MDR), and the Lucena cell line, resistant to chemotherapy (MDR) were investigated. The results suggest that both cell lines possess CD34+CD38- profiles of hematopoietic stem cell markers. The promoter regions of ABCB1, ABCG2 and ABCC1 genes contain binding sites for the Oct-4 transcripton factor, which is also considered a marker of tumor stem cells. Lucena cells showed an over-expression of the ABCB1 gene and a high expression of the Oct-4, ABCG2 and ABCC1 genes as compared to K562 cells.

Time-course expression of DNA repair-related genes in hepatocytes of zebrafish (Danio rerio) after UV-B exposure.

The objective of this study was to evaluate the time-course effects of UV-B exposure on expression of genes involved in the DNA repair system of zebrafish (Danio rerio) hepatocytes, a highly competent species in terms of damage repair induced by UV radiation. For gene expression analysis (RT-PCR), cells were exposed to 23.3 mJ cm(-2) UV-B, which was the dose that affected viable cell number (reduction of 30% when compared with the control group) and produced no visual alteration on cell morphology. The early response observed (6 h) showed induction in the expression of the CDKI gene (cyclin-dependent kinase inhibitor) and genes related to DNA damage repair (mainly XPC and DDB2), while the late response observed (24 h) was more related to up-regulation of p53 and genes involved in cell cycle arrest (gadd45a, cyclinG1). In all times analyzed, the anti-apoptotic gene Bcl-2 was down-regulated. Another interesting result observed was the up-regulation of the Apex-1 gene after UV-B exposure, which could indicate the induction of oxidative lesions in the DNA molecule. In conclusion, these results demonstrate an activation of the DNA repair system in hepatocytes of zebrafish exposed to UV-B radiation, mainly involving the participation of p53.

Toxicological responses in Laeonereis acuta (annelida, polychaeta) after arsenic exposure.

Several environmental pollutants, including metals, can induce oxidative stress. So, the objective of this study was to evaluate the effects of arsenic (As(III), as As(2)O(3)) on the antioxidant responses in the polychaete Laeonereis acuta. Worms were exposed to two environmentally relevant concentrations of As, including the highest previously allowed by Brazilian legislation (50 microg As/l). A control group was kept in saline water (10 per thousand) without added metal. It was observed that: (1) a peak concentration of lipid peroxide was registered after 2 days of exposure to 50 microg As/l (61+/-3.2 nmol CHP/g wet weight) compared to the control group (43+/-4.5 nmol CHP/g wet weight), together with a lowering of the activity of the antioxidant enzyme catalase (-47 and -48%, at 50 or 500 microg As/l respectively) and a higher superoxide dismutase activity (+305% at 50 microg As/l with respect to the control group); (2) a lower conjugation capacity through glutathione-S-transferase activity was observed after 7 days of exposure to 50 microg As/l (-48% compared to the control group); (3) a significant increase in As concentration was verified after 1 week of exposure to both As concentrations (50 and 500 microg/l); (4) worms exposed to As showed a limited accumulation of related methylated As species and the levels of non-toxic As species like arsenobetaine (AsB) and arsenocholine (AsC) remained unchanged during the exposure period when compared with the controls. Overall, it can be concluded that As interfered in the antioxidant defense system of L. acuta, even at low concentrations (50 microg/l) that Brazilian legislation previously considered safe. The fact that worms exposed to As showed high levels of methylated As species indicates the methylation capability of L. acuta, although the high levels of inorganic As suggest that not all the administered As(III) (as As(2)O(3)) is completely removed or biotransformed after 7 days of exposure.

cDNA cloning and expression analysis of the catalytic subunit of glutamate cysteine ligase gene in an annelid polychaete after cadmium exposure: a potential tool for pollution biomonitoring.

The enzyme glutamate cysteine ligase (GCL) is the rate-limiting enzyme in glutathione (GSH) synthesis and is formed by a catalytic (GCLC) and a modulatory subunit (GCLM). Some studies have demonstrated that environmental pollutants can regulate the expression of these subunits. Despite the importance of these genes in toxicological responses, no sequences are available for the GCL subunits in annelids. The present study reports, for the first time, the cDNA sequence for the GCLC in an annelid species, the polychaete Laeonereis acuta (Nereididae). The deduced amino acid sequence of L. acuta GCLC showed homology with other animal species, and was used to infer a phylogenetic tree with GCLC amino acid sequences from other taxonomic groups. Exposure to cadmium (100 and 1000 microg Cd/L) during 14 days augmented the level of L. acuta GCLC transcripts in a dose-dependent manner. These gene expression results can be related to the known cadmium effect on GSH depletion. Since a number of contaminants can also exert their toxicity through this mechanism, GCLC gene expression might be applied not only for cadmium biomonitoring, but also for a wide range of contaminants that directly or indirectly promote the same effect in the cellular GSH content.

Antioxidant properties of the mucus secreted by Laeonereis acuta (Polychaeta, Nereididae): a defense against environmental pro-oxidants?

Polychaeta species like Laeonereis acuta (Nereididae) usually secrete great amounts of mucus that wrap the animal inside. Taking into account that fungi action in the sediment and UV radiation acting on dissolved organic matter in the water produces reactive oxygen species (ROS) like hydrogen peroxide (H(2)O(2)), it was considered that the mucus secretion could represent an antioxidant defense against environmental ROS. Antioxidant enzymes (catalase-CAT; superoxide dismutase-SOD; glutathione peroxidase-GPx and glutathione-S-transferase-GST) and total antioxidant capacity (TOSC) were determined in worms and mucus secretion. Higher (p<0.05) CAT, GPx and TOSC values were registered in mucus samples respect worms, SOD activity was similar (p>0.05) in both kind of samples, and absence of GST activity was observed in mucus samples, suggesting absence of catalyzed phase II reactions. In assays conducted with hepatoma cell lines exposed to H(2)O(2), it was verified that: (1) mucus co-exposure significantly (p<0.05) lowered DNA damage induced by H(2)O(2); (2) ROS production was significantly (p<0.05) reduced when cells were exposed simultaneously with mucus samples and H(2)O(2) respect H(2)O(2) alone. It can be concluded that the mucus production contributes substantially to the antioxidant defense system of the worm against environmental ROS through the interception or degradation of H(2)O(2), peroxyl and hydroxyl radicals.