Cylindrospermopsin exposure promotes redox unbalance and tissue damage in the liver of Poecilia reticulata, a neotropical fish species.

There is a growing concern regarding the adverse risks exposure to cylindrospermopsin (CYN) might exert on animals and humans. However, data regarding the toxicity of this cyanotoxin to neotropical fish species are scarce. Using the fish species Poecilia reticulata, the influence of CYN concentrations equal to and above the tolerable for drinking water may produce on liver was determined by assessing biomarkers of antioxidant defense mechanisms and correlated to qualitative and semiquantitative histopathological observations. Adult females were exposed to 0.0 (Control); 0.5, 1 and 1.5 µg/L pure CYN for 24 or 96 hr, in triplicate. Subsequently the livers were extracted for biochemical assays and histopathological evaluation. Catalase (CAT) activity was significantly increased only by 1.5 µg/L CYN-treatment, at both exposure times. Glutathione -S-transferase (GST) activity presented a biphasic response for both exposure times. It was markedly decreased after exposure by 0.5 µg/L CYN treatment but significantly elevated by 1.5 µg/L CYN treatment. All CYN treatments produced histopathological alterations, as evidenced by hepatocyte cords degeneration, steatosis, inflammatory infiltration, melanomacrophage centers, vessel congestion, and areas with necrosis. Further, an IORG >35 was achieved for all treatments, indicative of the presence of severe histological alterations in P. reticulata hepatic parenchyma and stroma. Taken together, data demonstrated evidence that CYN-induced hepatotoxicity in P. reticulata appears to be associated with an imbalance of antioxidant defense mechanisms accompanied by histopathological liver alterations. It is worthy to note that exposure to low environmentally-relevant CYN concentrations might constitute a significant risk to health of aquatic organisms.

Importancia biológica de la microcistina en aguas de riego y alternativas en su detección y degradación / Biological importance of microcistin in irrigation and alternative waters in its detection and degradation

Introducción: Las cianobacterias son microrganismos fotosintéticos, con capacidad de sintetizar una gran diversidad de metabolitos secundarios de interés para la industria, pero también han llamado la atención en las últimas décadas las toxinas denominas cianotoxinas, metabolitos que causan distintas alternaciones fisiológicas hasta llegar ocasionar la muerte de diferentes especies. Metodología: La determinación del estado de arte para el tema de cianobacterias se basó en una búsqueda bibliográfica en la base de datos especializada como Elservier, Springer, Google académico y MDPI basadas en palabras clave en español e inglés "microcistinas", "degradación de MC" y "cuantificación y detección de MC". Resultados: En la presente revisión considera dos áreas de caracterización de la microcistinas (MCs) las propiedades fisicoquímicas y propiedades biológicas, para entender su comportamiento e importancia tóxica en los sembradíos agrícolas y en la salud humana. Además de comprender alternativas para su degradación, por métodos fisicoquímicos como fotocatálisis y la gradación biológica por bacterias. Finalmente se mencionará algunos métodos actuales y en desarrollo, para la detección y cuantificación de estas MCs en ambientes acuáticos. Conclusiones: Las MCs tienen el potencial contaminar fuentes de agua como ríos y lagunas, causando daños a la salud humana y a las plantas agrícolas, tienen la capacidad de tolerar distintos cambios drásticos en factores fisicoquímicos y biológicos. Entre las alternativas reportadas la degradación bacteriana promete ser la más confiable. Finalmente, entre los distintos métodos para la detección de MCs, entre los métodos más aplicados son los inmunoensayos, debido a su versatilidad y estabilidad del método.

Introduction: Cyanobacteria are photosynthetic microorganisms, with the capacity to synthesize a great diversity of secondary metabolites of interest to the industry, but toxins called cyanotoxins have also attracted attention in recent decades, metabolites that cause different physiological alterations until they cause the death of different species. Methodology: The determination of the state of the art for the subject of cyanobacteria was based on a bibliographic search in specialized databases such as Elservier, Springer, Google Scholar and MDPI based on keywords in Spanish and English "microcystins", "MC degradation " and "quantification and detection of MC". Results: In the present review, two areas of characterization of microcystins are considered: the physicochemical properties and biological properties of microcystins (MC), to understand their behavior and toxic importance in agricultural crops and in human health.In addition to understanding alternatives for their degradation, by physicochemical methods such as photocatalysis and biological grading by bacteria.Finally, some current and developing methods will be mentioned for the detection and quantification of these MCs in aquatic environments. Conclusions: MCs have the potential to contaminate water sources such as rivers and lagoon, causing damage to human health and agricultural plants, they have the ability to tolerate different drastic changes in physicochemical and biological factors. Among the reported alternatives, bacterial degradation promises to be the most reliable. Finally, among the different methods for the detection of MCs, among the most applied methods are immunoassays, due to their versatility and stability of the method.

Microcystin drives the composition of small-sized bacterioplankton communities from a coastal lagoon.

Cyanobacterial blooms affect biotic interactions in aquatic ecosystems, including those involving heterotrophic bacteria. Ultra-small microbial communities are found in both surface water and groundwater and include diverse heterotrophic bacteria. Although the taxonomic composition of these communities has been described in some environments, the involvement of these small cells in the fate of environmentally relevant molecules has not been investigated. Here, we aimed to test if small-sized microbial fractions from a polluted urban lagoon were able to degrade the cyanotoxin microcystin (MC). We obtained cells after filtration through 0.45 as well as 0.22 µm membranes and characterized the morphology and taxonomic composition of bacteria before and after incubation with and without microcystin-LR (MC-LR). Communities from different size fractions (< 0.22 and < 0.45 µm) were able to remove the dissolved MC-LR. The originally small-sized cells grew during incubation, as shown by transmission electron microscopy, and changed in both cell size and morphology. The analysis of 16S rDNA sequences revealed that communities originated from < 0.22 and < 0.45 µm fractions diverged in taxonomic composition although they shared certain bacterial taxa. The presence of MC-LR shifted the structure of < 0.45 µm communities in comparison to those maintained without toxin. Actinobacteria was initially dominant and after incubation with MC-LR Proteobacteria predominated. There was a clear enhancement of taxa already known to degrade MC-LR such as Methylophilaceae. Small-sized bacteria constitute a diverse and underestimated fraction of microbial communities, which participate in the dynamics of MC-LR in natural environments.

Cylindrospermopsin impairs tubular transport function in kidney cells LLC-PK1.

Cylindrospermopsin (CYN) has been involved in cases of poisoning in humans following ingestion. Studies have demonstrated that the kidney is the most affected organ. CYN exposure leads to low-molecular-weight proteinuria and increased excretions of the tubular enzymes in mice, suggesting the damage caused by CYN is mainly tubular. However, the mechanism involved in CYN nephrotoxicity remains unknown. Thus, in order to evaluate the effects of CYN exposure (0.1, 0.5 and 1.0 µg/mL) on tubular renal cells LLC-PK1 distinct mechanisms were analyzed by assessing cell death using flow cytometry, albumin uptake by fluorescence analysis, Na+/K+-ATPase activity by a colorimetric method, RT-qPCR of genes related to tubular transport and function as well as internalization of CYN by ELISA. In this study, CYN was found to induce necrosis in all concentrations. CYN also decreased albumin uptake as well as downregulated megalin and dab2 expression, both proteins involved in albumin endocytosis process. Moreover, CYN appears to be internalized by renal tubular cells through a receptor-mediated endocytosis. Finally, the present study demonstrates that CYN is responsible for disrupting tubular cell transport and function in LLC-PK1 cells.

DNA damage induced by cylindrospermopsin on different tissues of the biomonitor fish Poecilia reticulata.

The cyanotoxin cylindrospermopsin (CYN) is the second biggest cause of poisoning worldwide, both in humans and animals. Although CYN primarily affects the aquatic environments and can be absorbed in fishes by multiple routes, data reporting its toxicity and mechanism of action are still scarce in this group. Using P. reticulata as model species, it was evaluated whether CYN promotes mutagenic and genotoxic effects in different fish target tissues. Adult females were exposed in a static way to 0 (control), 0.5, 1.0, and 1.5 µg L-1 of pure CYN for 24 and 96 hours. For the first time, DNA damage was detected in fish brain after CYN exposition. In brain cells, a concentration-response DNA damage was observed for both exposure times, suggesting a direct or indirect action of CYN in neurotoxicity. For the liver cells, 96 hours caused an increase in DNA damage, as well the highest percentage of DNA in the tail was reached when used 1.5 µg L-1 of CYN. In peripheral blood cells, an increase in DNA damage was observed for all tested concentrations after 96 hours. In erythrocytes, micronuclei frequency was higher at 1.5 µg L-1 treatment while the erythrocyte nuclear abnormalities (ENA) frequency was significantly higher even at the lowest CYN concentration. Such data demonstrated that acute exposition to CYN promotes genotoxicity in the brain, liver, and blood cells of P. reticulata, as well mutagenicity in erythrocytes. It rises an alert regarding to the toxic effects of CYN for aquatic organisms as well as for human health.

Overview of toxic cyanobacteria and cyanotoxins in Ibero-American freshwaters: Challenges for risk management and opportunities for removal by advanced technologies.

The increasing occurrence of cyanobacterial blooms worldwide represents an important threat for both the environment and public health. In this context, the development of risk analysis and management tools as well as sustainable and cost-effective treatment processes is essential. The research project TALGENTOX, funded by the Ibero-American Science and Technology Program for Development (CYTED-2019), aims to address this ambitious challenge in countries with different environmental and social conditions within the Ibero-American context. It is based on a multidisciplinary approach that combines ecology, water management and technology fields, and includes research groups from Chile, Colombia, Mexico, Peru and Spain. In this review, the occurrence of toxic cyanobacteria and cyanotoxins in freshwaters from these countries are summarized. The presence of cyanotoxins has been confirmed in all countries but the information is still scarce and further monitoring is required. In this regard, remote sensing or metagenomics are good alternatives at reasonable cost. The risk management of freshwaters from those countries considering the most frequent uses (consumption and recreation) has been also evaluated. Only Spain and Peru include cyanotoxins in its drinking water legislation (only MC-LR) and thus, there is a need for regulatory improvements. The development of preventive strategies like diminishing nutrient loads to aquatic systems is also required. In the same line, corrective measures are urgently needed especially in drinking waters. Advanced Oxidation Processes (AOPs) have the potential to play a major role in this scenario as they are effective for the elimination of most cyanotoxins classes. The research on the field of AOPs is herein summarized considering the cost-effectiveness, environmental character and technical applicability of such technologies. Fenton-based processes and photocatalysis using solar irradiation or LED light represent very promising alternatives given their high cost-efficiency. Further research should focus on developing stable long-term operation systems, addressing their scale-up.

Copepod Prey Selection and Grazing Efficiency Mediated by Chemical and Morphological Defensive Traits of Cyanobacteria.

Phytoplankton anti-grazer traits control zooplankton grazing and are associated with harmful blooms. Yet, how morphological versus chemical phytoplankton defenses regulate zooplankton grazing is poorly understood. We compared zooplankton grazing and prey selection by contrasting morphological (filament length: short vs. long) and chemical (saxitoxin: STX- vs. STX+) traits of a bloom-forming cyanobacterium (Raphidiopsis) offered at different concentrations in mixed diets with an edible phytoplankton to a copepod grazer. The copepod selectively grazed on the edible prey (avoidance of cyanobacteria) even when the cyanobacterium was dominant. Avoidance of the cyanobacterium was weakest for the "short STX-" filaments and strongest for the other three strains. Hence, filament size had an effect on cyanobacterial avoidance only in the STX- treatments, while toxin production significantly increased cyanobacterial avoidance regardless of filament size. Moreover, cyanobacterial dominance reduced grazing on the edible prey by almost 50%. Results emphasize that the dominance of filamentous cyanobacteria such as Raphidiopsis can interfere with copepod grazing in a trait specific manner. For cyanobacteria, toxin production may be more effective than filament size as an anti-grazer defense against selectively grazing zooplankton such as copepods. Our results highlight how multiple phytoplankton defensive traits interact to regulate the producer-consumer link in plankton ecosystems.

Study of cyanotoxin degradation and evaluation of their transformation products in surface waters by LC-QTOF MS.

In the present work, the degradation of three cyanotoxins from the hepatotoxins group was investigated under laboratory-controlled experiments in water samples. Surface waters spiked with microcystin-LR (MC-LR), nodularin (NOD) and cylindrospermopsin (CYN) were subjected to hydrolysis, chlorination and photo-degradation, under both sunlight (SL) and ultraviolet (UV) radiation. A total of 12 transformation products (TPs) were detected and tentatively identified by liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (LC-QTOF MS). These comprised: 6 chlorination TPs (3 from CYN and 3 from MC-LR, 2 isomers); 4 UV TPs (all from CYN); and 2 sunlight TPs (one isomer from MC-LR and another from NOD). No TPs were observed under hydrolysis conditions. The chemical structures for all TPs were tentatively proposed based on the accurate-mass QTOF MS full-spectra. Analysis of real-world samples collected from the Peñol reservoir (Antioquia, Colombia) revealed the presence of MC-LR and CYN as well as a sunlight TP identified in the laboratory experiments. Data presented in this article will assist further research on TPs potentially formed in future tertiary degradation processes applied for the removal of organic micro-pollutants in water; as well as improving available knowledge on the toxic implications of cyanobacterial toxins TPs in surface waters.

Microcystis aeruginosa and microcystin-LR removal by household slow sand filters operating in continuous and intermittent flows.

A household slow sand filter (HSSF) is a widely used water treatment technology recognized as one of the most effective and sustainable in reducing waterborne diseases. However, there is a lack of knowledge concerning its behaviour in the presence of cyanobacteria and cyanotoxins. In this context, the study aimed to evaluate HSSF ability to remove Microcystis aeruginosa cells (stain BB005) and microcystin-LR from water, among other parameters, when operated under continuous (C-HSSF) and intermittent (I-HSSF) flows. CHSSF was operated at a constant filtration rate (1.22 m3 m-2 day-1), while I-HSSF was operated at a variable filtration rate (starting at 2.95 m3 m-2 day-1 and finishing at zero). Each filter produced 60 L day-1. The influence of the pause period was also tested in the I-HSSF. The water from the study was prepared by inoculating M. aeruginosa culture in water from a well to a final cell density of ± 1 × 105 cells mL-1. M. aeruginosa removal rates were 2.39 ±â€¯0.34 log and 2.01 ±â€¯0.43 log by CHSSF and I-HSSF, respectively. Microcystin-LR concentration in studied water was 5.55 µg L-1, and both filters produced filtered water with microcystin concentrations below 1.0 µg L-1, the maximum value recommended by the World Health Organization (WHO), for most of the samples. Turbidity and apparent colour were also within WHO guidelines. Filters operating with different flow regimes and distinct residence times did not statistically influence treatment efficiencies. Both filters showed promising results in the M. aeruginosa and microcystin-LR removals from water; nevertheless, more research is needed to understand the mechanisms involved in the reduction of both cyanobacteria and cyanotoxin through household slow sand filtration.

Genetic toxicity of water contaminated by microcystins collected during a cyanobacteria bloom.

Microcystin-LR (MCLR) is a toxin mainly produced by Microcystis aeruginosa, cyanobacteria most commonly found in eutrophic environments. Cyanobacteria blooms have affected Salto Grande reservoir (Americana, State of São Paulo/Brazil) for several decades, often observed during periods of drought. In this study, the genotoxic effects of MCLR (95% purity) and water samples contaminated by this toxin were evaluated during cyanobacteria bloom using assays with the test organism Allium cepa. The results showed genotoxic action for pure microcystin and cytotoxic, genotoxic and mutagenic action for water samples collected during flowering. Chromosomal aberration assays have shown that MCLR induces chromosomal breaks that persist in the daughter cells as MN. Therefore, it is possible to infer a clastogenic action for this toxin. The MCLR present in the environmental samples was shown to be more cytogenotoxic for the cells than the different concentrations tested in this study with the pure substance. This amplified toxic action can be related to a synergistic effect between the MCLR and other compounds present in the environmental samples. The genotoxicity studies with MCLR show inconsistent and inconclusive results, so this toxin needs to be better investigated in order to obtain further information about the action mode of it is on the biological system.

Cylindrospermopsin effects on cell viability and redox milieu of Neotropical fish Hoplias malabaricus hepatocytes.

Cylindrospermopsin (CYN) is a cyanotoxin that is cytotoxic to a wide variety of cells, particularly to the hepatocytes. In this study, the toxic effects of purified CYN were investigated in primary cultured hepatocytes of Neotropical fish Hoplias malabaricus. After isolation, attachment, and recovery for 72 h, the cells were exposed for 72 h to 0, 0.1, 1.0, 10, and 100 µg l-1 of CYN. Then, cell viability and a set of oxidative stress biomarker responses were determined. Catalase, superoxide dismutase, glucose-6-phosphate dehydrogenase, and glutathione S-transferase activities were not affected by exposure to CYN. Concentration-dependent decrease of glutathione reductase activity occurred for most CYN-exposed groups, whereas non-protein thiol content increased only for the highest CYN concentration. Lipid peroxidation, protein carbonylation, and DNA damage levels were not altered, but reactive oxygen species levels increased in the cells exposed to the highest concentration of CYN. Cell viability decreased in all the groups exposed to CYN. Thus, CYN may cause a slight change in redox balance, but it is not the main cause of cell death in H. malabaricus hepatocytes.

Degradation of cyanotoxins (microcystin) in drinking water using photoelectrooxidation / Degradação de cianotoxinas (microcistinas) em água potável aplicando fotoeletrooxidação

The discharge of sewage and industrial effluents containing high concentrations of pollutants in water bodies increases eutrophication. Cyanobacteria, some of the organisms whose growth is promoted by high nutrient concentrations, are resistant and produce several types of toxins, known as cyanotoxins, highly harmful to human beings. Current water treatment systems for the public water supply are not efficient in degradation of toxins. Advanced oxidation processes (AOP) have been tested for the removal of cyanotoxins, and the results have been positive. This study examines the application of photoelectrooxidation in the degradation of cyanotoxins (microcystins). The performance of the oxidative processes involved was evaluated separately: Photocatalysis, Electrolysis and Photoelectrooxidation. Results showed that the electrical current and UV radiation were directly associated with toxin degradation. The PEO system is efficient in removing cyanotoxins, and the reduction rate reached 99%. The final concentration of toxin was less than 1 µg/L of microcystin in the treated solution.

A descarga de esgotos e efluentes industriais contendo altas concentrações de poluentes nos corpos d'água aumenta a eutrofização. As cianobactérias, são organismos cujo crescimento é promovido por concentrações elevadas de nutrientes, são resistentes e produzem vários tipos de toxinas conhecidas, como cianotoxinas, altamente prejudiciais para os seres humanos. Os sistemas atuais de tratamento de água para o abastecimento público de água não são eficientes na degradação destas toxinas. Processos oxidativos avançados (POA) foram testados para a remoção de cianotoxinas, e os resultados têm sido positivos. Este estudo avalia o processo de fotoeletrooxidação (FEO) na degradação de cianotoxinas (microcistinas). Foi avaliado o desempenho dos processos envolvidos separadamente: fotocatalisis, eletrólise e fotoeletrooxidação. Os resultados mostram que a potencia da radiação UV e da corrente elétrica estão diretamente associados com a degradação de toxinas. O sistema de FEO é eficiente na remoção de cianotoxinas e a redução foi de 99%. A concentração final de toxina foi inferior a 1 g / L de microcistina na solução tratada.

Degradation of cyanotoxins (microcystin) in drinking water using photoelectrooxidation / Degradação de cianotoxinas (microcistinas) em água potável aplicando fotoeletrooxidação

The discharge of sewage and industrial effluents containing high concentrations of pollutants in water bodies increases eutrophication. Cyanobacteria, some of the organisms whose growth is promoted by high nutrient concentrations, are resistant and produce several types of toxins, known as cyanotoxins, highly harmful to human beings. Current water treatment systems for the public water supply are not efficient in degradation of toxins. Advanced oxidation processes (AOP) have been tested for the removal of cyanotoxins, and the results have been positive. This study examines the application of photoelectrooxidation in the degradation of cyanotoxins (microcystins). The performance of the oxidative processes involved was evaluated separately: Photocatalysis, Electrolysis and Photoelectrooxidation. Results showed that the electrical current and UV radiation were directly associated with toxin degradation. The PEO system is efficient in removing cyanotoxins, and the reduction rate reached 99%. The final concentration of toxin was less than 1 µg/L of microcystin in the treated solution..(AU)

A descarga de esgotos e efluentes industriais contendo altas concentrações de poluentes nos corpos d'água aumenta a eutrofização. As cianobactérias, são organismos cujo crescimento é promovido por concentrações elevadas de nutrientes, são resistentes e produzem vários tipos de toxinas conhecidas, como cianotoxinas, altamente prejudiciais para os seres humanos. Os sistemas atuais de tratamento de água para o abastecimento público de água não são eficientes na degradação destas toxinas. Processos oxidativos avançados (POA) foram testados para a remoção de cianotoxinas, e os resultados têm sido positivos. Este estudo avalia o processo de fotoeletrooxidação (FEO) na degradação de cianotoxinas (microcistinas). Foi avaliado o desempenho dos processos envolvidos separadamente: fotocatalisis, eletrólise e fotoeletrooxidação. Os resultados mostram que a potencia da radiação UV e da corrente elétrica estão diretamente associados com a degradação de toxinas. O sistema de FEO é eficiente na remoção de cianotoxinas e a redução foi de 99%. A concentração final de toxina foi inferior a 1 g / L de microcistina na solução tratada.(AU)

Degradation of cyanotoxins (microcystin) in drinking water using photoelectrooxidation

p>The discharge of sewage and industrial effluents containing high concentrations of pollutants in water bodies increases eutrophication. Cyanobacteria, some of the organisms whose growth is promoted by high nutrient concentrations, are resistant and produce several types of toxins, known as cyanotoxins, highly harmful to human beings. Current water treatment systems for the public water supply are not efficient in degradation of toxins. Advanced oxidation processes (AOP) have been tested for the removal of cyanotoxins, and the results have been positive. This study examines the application of photoelectrooxidation in the degradation of cyanotoxins (microcystins). The performance of the oxidative processes involved was evaluated separately: Photocatalysis, Electrolysis and Photoelectrooxidation. Results showed that the electrical current and UV radiation were directly associated with toxin degradation. The PEO system is efficient in removing cyanotoxins, and the reduction rate reached 99%. The final concentration of toxin was less than 1 µg/L of microcystin in the treated solution. /p>

p>A descarga de esgotos e efluentes industriais contendo altas concentrações de poluentes nos corpos d'água aumenta a eutrofização. As cianobactérias, são organismos cujo crescimento é promovido por concentrações elevadas de nutrientes, são resistentes e produzem vários tipos de toxinas conhecidas, como cianotoxinas, altamente prejudiciais para os seres humanos. Os sistemas atuais de tratamento de água para o abastecimento público de água não são eficientes na degradação destas toxinas. Processos oxidativos avançados (POA) foram testados para a remoção de cianotoxinas, e os resultados têm sido positivos. Este estudo avalia o processo de fotoeletrooxidação (FEO) na degradação de cianotoxinas (microcistinas). Foi avaliado o desempenho dos processos envolvidos separadamente: fotocatalisis, eletrólise e fotoeletrooxidação. Os resultados mostram que a potencia da radiação UV e da corrente elétrica estão diretamente associados com a degradação de toxinas. O sistema de FEO é eficiente na remoção de cianotoxinas e a redução foi de 99%. A concentração final de toxina foi inferior a 1 g / L de microcistina na solução tratada. /p>

Biochemical and genetic alterations in the freshwater neotropical fish Prochilodus lineatus after acute exposure to Microcystis aeruginosa

Microcystins are secondary metabolites produced by different species of cyanobacteria, such as Microcystis aeruginosa (MA). In this study, the biochemical and genetic effects of lyophilized MA were evaluated in the neotropical fish Prochilodus lineatus exposed to 1 or 2 mg L-1 lyophilized MA (treated group) or only water (control group) in static toxicity tests for 24 and 96 h. The gills and liver were used in the analysis of biotransformation enzymes and antioxidant defenses, blood and gill cells in genetic analysis and in brain and muscle it was determined the activity of acetylcholinesterase (AChE). The results showed the biotransformation pathway activation due to the increase in hepatic CYP1A and in branchial and hepatic glutathione S-transferase (GST). The antioxidant defense proved to be greatly affected by MA exposure leading to changes, both in gills and liver, in the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and in the content of tripeptide glutathione (GSH). Lipid peroxidation was not detected, but damage to DNA molecule was observed in blood cells. In conclusion, it can be state the lyophilized MA is able to promote changes in the biochemical and genetic parameters of P. lineatus.

As microcistinas são metabólitos secundários produzidos por diferentes espécies de cianobactérias, como a Microcystis aeruginosa (MA). Neste estudo, os efeitos bioquímicos e genéticos de liofilizado de MA foram avaliados para juvenis da espécie de peixe Neotropical Prochilodus lineatus expostos a 1 ou 2 mg L-1 de liofilizado de MA (grupo tratado) ou apenas à água (grupo controle), em testes de toxicidade estáticos, durante 24 e 96 h. As brânquias e o fígado foram usados para as análises das enzimas de biotransformação e defesas antioxidantes, células do sangue e das brânquias para análises genéticas e no cérebro e músculo foi determinada a atividade da acetilcolinesterase (AChE). Os resultados mostraram ativação da via de biotransformação devido ao aumento da atividade da CYP1A hepática e da atividade da glutationa S-transferase (GST) hepática e branquial. As defesas antioxidantes foram muito afetadas pela exposição a MA levando a alterações, tanto no fígado como nas brânquias, na atividade da superóxido dismutase (SOD), da catalase (CAT), da glutationa peroxidase (GPx), glutationa redutase (GR) e no conteúdo do tripeptídeo glutationa (GSH). Apesar dessas alterações a peroxidação lipídica não foi detectada em nenhum dos tecidos, mas danos na molécula de DNA foram observados nas células do sangue. Em conclusão, pode-se afirmar que o liofilizado de MA é capaz de promover alterações em parâmetros bioquímicos e genéticos de P. lineatus.

Biochemical and genetic alterations in the freshwater neotropical fish Prochilodus lineatus after acute exposure to Microcystis aeruginosa

Microcystins are secondary metabolites produced by different species of cyanobacteria, such as Microcystis aeruginosa (MA). In this study, the biochemical and genetic effects of lyophilized MA were evaluated in the neotropical fish Prochilodus lineatus exposed to 1 or 2 mg L-1 lyophilized MA (treated group) or only water (control group) in static toxicity tests for 24 and 96 h. The gills and liver were used in the analysis of biotransformation enzymes and antioxidant defenses, blood and gill cells in genetic analysis and in brain and muscle it was determined the activity of acetylcholinesterase (AChE). The results showed the biotransformation pathway activation due to the increase in hepatic CYP1A and in branchial and hepatic glutathione S-transferase (GST). The antioxidant defense proved to be greatly affected by MA exposure leading to changes, both in gills and liver, in the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and in the content of tripeptide glutathione (GSH). Lipid peroxidation was not detected, but damage to DNA molecule was observed in blood cells. In conclusion, it can be state the lyophilized MA is able to promote changes in the biochemical and genetic parameters of P. lineatus.(AU)

As microcistinas são metabólitos secundários produzidos por diferentes espécies de cianobactérias, como a Microcystis aeruginosa (MA). Neste estudo, os efeitos bioquímicos e genéticos de liofilizado de MA foram avaliados para juvenis da espécie de peixe Neotropical Prochilodus lineatus expostos a 1 ou 2 mg L-1 de liofilizado de MA (grupo tratado) ou apenas à água (grupo controle), em testes de toxicidade estáticos, durante 24 e 96 h. As brânquias e o fígado foram usados para as análises das enzimas de biotransformação e defesas antioxidantes, células do sangue e das brânquias para análises genéticas e no cérebro e músculo foi determinada a atividade da acetilcolinesterase (AChE). Os resultados mostraram ativação da via de biotransformação devido ao aumento da atividade da CYP1A hepática e da atividade da glutationa S-transferase (GST) hepática e branquial. As defesas antioxidantes foram muito afetadas pela exposição a MA levando a alterações, tanto no fígado como nas brânquias, na atividade da superóxido dismutase (SOD), da catalase (CAT), da glutationa peroxidase (GPx), glutationa redutase (GR) e no conteúdo do tripeptídeo glutationa (GSH). Apesar dessas alterações a peroxidação lipídica não foi detectada em nenhum dos tecidos, mas danos na molécula de DNA foram observados nas células do sangue. Em conclusão, pode-se afirmar que o liofilizado de MA é capaz de promover alterações em parâmetros bioquímicos e genéticos de P. lineatus.(AU)

Microcistina en plantas de tratamiento de agua para consume humano en un ambiente tropical: el Area Metropolitana de Costa Rica / Microcystin in plants that treat water for human consumption in a tropical environment: the Metropolitan Area of Costa Rica

We measured microcystin levels in water of the Metropolitan Area of Costa Rica by competitive inhibition ELISA and we quantified total coliforms, fecal coliforms, Escherichia coli (by a Most Probable Number method) and aerobic count. We wanted to identify any cyanotoxin correlation with these parameters, as a public health risk. We sampled in the rainy season of 2003 (April-October) and in the dry season of 2004 (February-March) (30 samples/season). We sampled pre-treated, semi-treated and treated water. Microcystin levels < 0.5 ppb were found in the rainy season (and > 0.5 ppb in the dry season). Dry season levels exceeded World Health Organization limits (1.0 ppb). Cyanotoxins occurred in the Tres Rios plant. We did not find a correlation between these microbiologic parameters of water quality and microcystin levels in water.

Se midió la presencia de microcistina en el Área Metropolitana de Costa Rica por la técnica de ELISA de inhibición competitiva y se cuantificó coliformes totales, coliformes termotolerantes, Escherichia coli (por medio de la técnica Numero Más Probable) y recuento total aerobio. Se realizaron dos etapas de muestreo, una durante la estación lluviosa del 2003 (abril-octubre) y otra durante la estación seca del 2004 (febrero-marzo), cada una con 30 muestras de agua. Se muestreó agua pretratada, semitratada y tratada. Se determinaron niveles de microcistina < 0.5 ppb durante la estación lluviosa del 2003, mientras que durante la estación seca, se detectaron concentraciones de microcistina > 0.5 ppb. Se informó la presencia de cianotoxinas en la Planta de Tratamiento de Tres Ríos. No se establece correlación entre los parámetros microbiológicos de calidad del agua y las concentraciones de microcistina en el agua. Los estudios deberían considerar la diversidad y toxicidad de cianobacterias en estas plantas, los efectos del tratamiento, y presencia de otros microorganismos y sustancias (dióxido de carbono, fósforo, Nitrógeno), sobre la presencia, estructura y efecto de estas toxinas.

Hepatotoxicidade da cianotoxina microcistina / Hepatotoxicity of the microcystin cyanotoxin

Constitui interesse emergente em saúde pública avaliar a possibilidade de intoxicação humana por biotoxinas de algas cianofíceas, principalmente as hepatotoxinas do grupo das microcistinas. A microcistina, um heptapeptídeo monocíclico, é produzida principalmente pela cianobactéria Microcistis aeruginosa. São caracterizadas por alguns aminoácidos variáveis, dois deles com uma estrutura não usual que possuem importante papel na hepatotoxidade da microcistina. Apesar do acometimento humano atribuído as microcistinas incluírem gastroenterite, reações alérgicas ou irritativas, neurotoxicidade, o principal alvo da toxina é o fígado. Nos hepatócitos as microcistinas são carreadas pelo sistema transportador do ácido biliar, inibindo a atividade da proteína fosfatase no citoplasma. A inibição leva a mudanças morfológicas na membrana plasmática pela hiperfosforilação de citoqueratinas, e à atividade de promoção tumoral pelas proteínas hiperfosforiladas. Os métodos de detecção e quantificação de microcistinas no ambiente incluem a cromatografia líquida, o bioensaio em camundongos e os testes imunoenzimáticos. O último vem ganhando destaque pela praticidade e alta sensibilidade.

At public health, there is increasingly interest on evaluating the possibility of human intoxication by biotoxins from blue-green algae, mainly the hepatotoxins from the microcystin group. Microcystin, a monocyclic heptapeptide, is mainly produced by a cyanobacteria called Microcistis aeruginosa. It is characterized by a few variable amino acids, from which two of them have an unusual structure and play an important role in the hepatotoxicity of the microcystin. Although human illnesses include gastroenteritis, allergic or irritative reactions, and neurotoxicity, the main target of this toxin is the liver. Inside the hepatocytes, microcystins are carried by the transportation system of the bile acid, inhibiting the activity of the protein phosphatase in the cytoplasm. This inhibition causes a morphologic change in the plasmatic membrane because of the hyperphosphorylation of cytokeratins, and also the tumoral promotion by the hyperphosphorylated proteins. The techniques used in the detection and quantification of the microcystins in the environment include liquid chromatography, bioanalysis of mice, and immunoenzymatic tests using mono and polyclonal antibodies against those toxins. The latter has been remarked because of its practicality and its high sensibility.