Valéria Freitas de Magalhães

Microcystin contamination in fish from the Jacarepaguá Lagoon (Rio de Janeiro, Brazil): ecological implication and human health risk.

Chronic and subchronic toxicity from exposure to microcystins, cyclic peptide liver toxins from certain cyanobacteria, poses an important hazard, which has received little study. No in vivo information exists on accumulation and transfer of microcystin from the food chain to humans. This paper present results of a 3-year study that demonstrates bioaccumulation of microcystins by fish and potential rates of microcystin ingestion by humans. The study was carried out in a shallow coastal lagoon in the city of Rio de Janeiro (Jacarepaguá Lagoon). Fish (Tilapia rendalli) were collected every 2 weeks from August 1996 to November 1999. Microcystins were analyzed by HPLC in phytoplankton, fish liver and viscera while fish muscle tissue was analyzed by enzyme linked immunosorbant assay (ELISA). Phytoplankton samples, dominated by the genus Microcystis, were confirmed to contain microcystins as were fish livers, viscera and muscle tissue. During the entire study period, including times of low water bloom densities, fish muscle tissue contained concentrations of microcystins close to or above the recommended limit for human consumption (0.04 microg x kg(-1) day). Our findings demonstrate that microcystins can accumulate in fish tissue used for human consumption. Rates of ingestion routinely exceed the TDI guidelines as set by the WHO for drinking water. Appropriate epidemiology and risk assessment should be undertaken so that an acceptable TDI and appropriate risk management decisions can be made for human consumption of fish which are harvested from cyanobacterial blooms that contain cyanotoxins.

Trace metal accumulation by algae in Sepetiba Bay, Brazil.

Trace metal concentrations (Zn, Cd, Pb, Cr and Cu) were determined in some benthic algae from Sepetiba Bay, Brazil. This region has been modified by the increase of industrial activities during the last 30 years. Among species sampled the brown algae Padina gymnospora contained the greatest amount of Zn. The Bioconcentration Factor for Zn in P. gymnospora was found to be 10(4) under laboratory conditions. The absorption of (65)Zn depended on exposure time and increased with Zn concentrations in the medium. An insignificant amount of Zn desorbed from algae suggested a very high affinity with cellular binding sites. Zinc uptake by P. gymnospora is discussed by considering field and laboratory conditions.

A rapid bioassay for detecting saxitoxins using a Daphnia acute toxicity test.

Bioassays using Daphnia pulex and Moina micrura were designed to detect cyanobacterial neurotoxins in raw water samples. Phytoplankton and cyanotoxins from seston were analyzed during 15 months in a eutrophic reservoir. Effective time to immobilize 50% of the exposed individuals (ET50) was adopted as the endpoint. Paralysis of swimming movements was observed between approximately 0.5-3 h of exposure to lake water containing toxic cyanobacteria, followed by an almost complete recovery of the swimming activity within 24 h after being placed in control water. The same effects were observed in bioassays with a saxitoxin-producer strain of Cylindrospermopsis raciborskii isolated from the reservoir. Regression analysis showed significant relationships between ET50 vs. cell density, biomass and saxitoxins content, suggesting that the paralysis of Daphnia in lake water samples was caused by saxitoxins found in C. raciborskii. Daphnia bioassay was found to be a sensitive method for detecting fast-acting neurotoxins in natural samples, with important advantages over mouse bioassays.

Cellular responses of Prochilodus lineatus hepatocytes after cylindrospermopsin exposure

Cylindrospermopsin is a potent toxicant for eukaryotic cells produced by several cyanobacteria. Recently, primary hepatocyte cultures of Neotropical fish have been established, demonstrating to be a quite efficient in vitro model for cellular toxicology studies. In the current study, a protocol for culture of Prochilodus lineatus hepatocytes was established and utilized to investigate the cellular responses to purified cylindrospermopsin exposure. Hepatocytes were successfully dissociated with dispase, resulting in a cell yield of 6.36 × 10(7)cells g(-1) of liver, viability of 97% and attachment on uncoated culture flasks. For investigation of cylindrospermopsin effects, hepatocytes were dissociated, cultured during 96 h and exposed to three concentrations of the toxin (0.1, 1.0 or 10 μgl(-1)) for 72 h. Cylindrospermopsin exposure significantly decreased cell viability (0.1 and 1 μgl(-1)) and multixenobiotic resistance mechanism, MXR (all exposed groups), but increased reactive oxygen/nitrogen species levels (all exposed groups) and lipid peroxidation (10 μgl(-1)). On the other hand no significant alterations were observed for other biochemical biomarkers as 2GSH/GSSG ratio, protein carbonyl levels and DNA strand breaks or glutathione S-transferase and glucose 6-phosphate dehydrogenase activities. In conclusion, hepatocytes might be made sensitive to cylindrospermopsin, at least in part, due to reduction of xenobiotics and endobiotics efflux capacity by MXR. Additionally, the toxin exposure suggests important issues regarding hepatocytes survival at the lowest cylindrospermopsin concentrations.

Arsenic concentration in sediments near a metallurgical plant (Sepetiba Bay, Rio de Janeiro, Brazil)

The Engenho Inlet, situated in Sepetiba Bay, has been undergoing transformations since the 1960s. due to the installation of a zinc metallurgical industry. In the chemical production process, arsenic is used to purify ore. Results show that most samples are above natural arsenic levels (10 mg kg−1), reaching 360 mg kg−1 near the effluent in the strongly bound fraction and 140 mg kg−1 in the weakly bound fraction at a depth of 28 cm. In the middle of the Inlet the values were 260 mg kg− and 80 mg kg−1 in the strongly and weakly bound fractions at a depth of 20 cm. These results indicate arsenic remobilization towards the Sepetiba Bay. The high availability of this metalloid in the ecosystem can damage the local biota and probably the local population as well.

Single and combined effects of microcystin‐ and saxitoxin‐producing cyanobacteria on the fitness and antioxidant defenses of cladocerans

Cyanobacteria produce different toxic compounds that affect animal life, among them hepatotoxins and neurotoxins. Because cyanobacteria are able to produce a variety of toxic compounds at the same time, organisms may be, generally, subjected to their combined action. In the present study, we demonstrate the single and combined effects on cladocerans of cyanobacteria that produce microcystins (hepatotoxins) and saxitoxins (neurotoxins). Animals were exposed (either singly or combined) to 2 strains of cyanobacteria isolated from the same environment (Funil Reservoir, Rio de Janeiro, Brazil). The effects on clearance rate, mobility, survivorship, fecundity, population increase rate (r), and the antioxidant enzymes glutathione-S-transferase (GST) and catalase (CAT) were measured. Cladoceran species showed a variety of responses to cyanobacterial exposures, going from no effect to impairment of swimming movement, lower survivorship, fecundity, and general fitness (r). Animals ingested cyanobacteria in all treatments, although at lower rates than good food (green algae). Antioxidant defense responses were in accordance with fitness responses, suggesting that oxidative stress may be related to such effects. The present study emphasizes the need for testing combined actions of different classes of toxins, because this is often, and most likely, the scenario in a more eutrophic world with global climatic changes. Environ Toxicol Chem 2017;36:2689-2697.

Neurotoxins in a water supply reservoir: An alert to environmental and human health

ABSTRACT Reservoirs are important source of power generation, recreation, and water supply. Nevertheless, human activities have favored the bloom of toxic cyanobacteria in many reservoirs, which has resulted in environmental, social, and economic problems. This study aims to evaluate the water quality of a reservoir in South Brazil through the analysis of cyanobacteria and cyanotoxins PSTs (Paralytic Shellfish Toxins) and biomarkers of environmental contamination in fish. For this purpose, water samples and fish (Geophagus brasiliensis) (Perciformes: Cichlidae) were collected from September 2013 to May 2014. The fish G. brasiliensis were separated in two groups. The first one “site group” was euthanized after the sampling and their weight and length were measured. The blood, brain, muscle and liver were collected for chemical, biochemical and genetics biomarkers analysis. The second group “depuration group” was submitted to depuration experiment for 40 days in clean water. After that, the same procedures as for the first group were carried out. Cylindrospermopsis raciborskii was the dominant cyanobacteria found in the reservoir, and it showed a density above the recommended limit by Brazilian legislation of 20,000 cells/mL. Results showed that the fish accumulate PSTs in the Reservoir and these were not eliminated after 40 days. The biochemical and genotoxic biomarkers showed a significant difference between “site groups” and “depuration groups”, which suggests a recovery of the antioxidant system and a reduction of cellular damage after 40 days in clean water. In conjunction with results reported earlier by others, Alagados Reservoir, in South Brazil, appears to have a persistent contamination of cyanotoxins. Moreover, the mixture of contaminants which may be present in the water body can explain the seasonal differences in fish at the sampled points. HIGHLIGHTSCyanobacterial blooms in Alagados Reservoir.Cyanobacteria densities above the recommended limit.Fish accumulated cyanotoxins.Cyanobacterial blooms can cause social, economic and environmental problems.

The accumulation dynamics, elimination and risk assessment of paralytic shellfish toxins in fish from a water supply reservoir

Paralytic shellfish Toxins (PSTs) or saxitoxins are neurotoxins that block the neural transmission by binding to the voltage-gated sodium channels in the nerve cells. There are >50 analogues described, which could be biotransformed into a molecular form of greater or lesser toxicity. The Alagados Reservoir is used for water supply, and persistent cyanobacterial blooms as well as PSTs concentrations have been found in this water body since 2002. The aims of this study were to quantify the concentrations of PSTs in the water and fish samples from the Alagados Reservoir. In addition, we evaluated the elimination of PSTs for 90 days in fish and estimated the potential risk to human health. Water and fish samples were collected from the reservoir. For the water samples the phytoplankton and chemical analyses were carried out. Fish were divided into two sample times: Field Samples (FS) and Elimination Experiment Samples (EES), which were maintained for 90 days in filtered and dechlorinated water. For chemical analysis, the muscles of FS were collected on the fish sampling day and the muscles and feces of EES were collected at 7, 15, 30, 45, 60, 75 and 90 days. PSTs concentrations were present in water and fish samples, and they were estimated as a potential risk to humans; mainly for children. In addition, toxins were accumulated, biotransformed to other analogues and excreted by the fish. However, after 90 days, the toxins were still present in the water and fish muscle. Therefore, PSTs can remain for a long period in water, and fish can be a carrier of these neurotoxins. New approaches of monitoring and management are necessary in the actual global context of cyanobacteria and cyanotoxins.

Comparative bioaccumulation and effects of purified and cellular extract of cylindrospermopsin to freshwater fish Hoplias malabaricus

ABSTRACT Many tropical freshwater ecosystems are impacted by cyanobacteria blooms increasing the risk of cyanotoxins exposure to aquatic organisms while human populations may be exposed by eating fish, drinking water, or dermal swimming. However, few toxicological data are available on the influence of cyanobacteria blooms in particular, cylindrospermopsin (CYN) on Brazilian neotropical fish. A number of studies demonstrated the ability of CYN to bioaccumulate in freshwater organisms and consequently enter the human food chain. The aim of the current study was to examine the effects of CYN following single intraperitoneal injection (50 µg/kg) of purified CYN (CYNp) or aqueous extract of CYN-producing cyanobacteria extract (CYNex) after 7 or 14 days. Biomarkers such as histopathology (liver), oxidative stress (liver and brain), and acetylcholinesterase (AChE) activity (muscle and brain) were utilized in order to assess the influence of CYN on Hoplias malabaricus. In terms of AChE activity, administration of CYNex and CYNp both muscle and brains were used as target tissues. In brain an increase of glutathione S-transferase (GST) activity and lipid peroxidation (LPO) levels was noted suggesting an imbalance in redox cycling. The majority of biomarkers did not present significant alterations in liver, but an elevation in superoxide dismutase (SOD) and glucose 6 phosphate dehydrogenase (G6PDH) activities was found. Different profiles of GST activity were observed in both studied groups (CYNex and CYNp) while LPO (CYNex and CYNp) and protein carbonylation (PCO) (CYNp) levels increased after exposure to CYN. The incidence of necrosis, melanomacrophages centers, and free melanomacrophages were detected as evidence of cell death and immune responses. Nonprotein thiols (NPT) levels were not markedly affected in both exposed groups. Data demonstrated that in vivo exposure to CYN produced biochemical and morphological disturbances in liver and brain of H. malabaricus.

Depuration time and sublethal effects of microcystins in a freshwater fish from water supply reservoir

Microcystins (MCs) are hepatotoxins that have been considered to be a worldwide problem due the effects that they can cause to environmental and human health systems. The Iraí Reservoir, located in the South of Brazil, is used as a water supply and MCs concentrations have been reported in this ecosystem. This study aimed to determine the MCs concentrations in the Iraí Reservoir and to evaluate the MCs depuration time and the health of Geophagus brasiliensis using biomarkers. Water and fish samples were collected in the Iraí Reservoir from August 2015 to May 2016. Phytoplankton and chemical analyses were conducted using water samples and the fish were divided into two groups; the Immediate Group (IMM) and the Depuration Group (DEP). In the IMM group, the blood, liver, muscle, brain and gills were collected, in order to evaluate the genotoxic, biochemical and chemical biomarkers. The DEP group was used in the depuration experiment for 90 days, and after this period the fish were submitted to the same procedure as the IMM group. Our results suggested that fish accumulated MCs and it may have caused oxidative stress, neurotoxicity and molecular damage. Furthermore, MCs concentrations increased during the depuration time and it resulted in molecular damage over the first 30 days. After 90 days, the recovery of the antioxidant system occurred. The depuration started on the 15th day, however, the toxins were still present in the samples. Therefore, the effects and the persistence of MCs are a risk to environmental systems and human health.