RESUMO
Cyanobacteria inhabiting lotic environments have been poorly studied and characterized in Mexico, despite their potential risks from cyanotoxin production. This article aims to fill this knowledge gap by assessing the importance of benthic cyanobacteria as potential cyanotoxin producers in central Mexican rivers through: (i) the taxonomic identification of cyanobacteria found in these rivers, (ii) the environmental characterization of their habitats, and (iii) testing for the presence of toxin producing genes in the encountered taxa. Additionally, we introduce and discuss the use of the term "CyanoHAMs" for lotic water environments. Populations of cyanobacteria were collected from ten mountain rivers and identified using molecular techniques. Subsequently, these taxa were evaluated for genes producing anatoxins and microcystins via PCR. Through RDA analyses, the collected cyanobacteria were grouped into one of three categories based on their environmental preferences for the following: (1) waters with high ionic concentrations, (2) cold-temperate waters, or (3) waters with high nutrient enrichment. Populations from six locations were identified to genus level: Ancylothrix sp., Cyanoplacoma sp., and Oxynema sp. The latter was found to contain the gene that produces anatoxins and microcystins in siliceous rivers, while Oxynema tested positive for the gene that produces microcystins in calcareous rivers. Our results suggest that eutrophic environments are not necessarily required for toxin-producing cyanobacteria. Our records of Compactonostoc, Oxynema, and Ancylothrix represent the first for Mexico. Four taxa were identified to species level: Wilmottia aff. murrayi, Nostoc tlalocii, Nostoc montejanii, and Dichothrix aff. willei, with only the first testing positive using PCR for anatoxin and microcystin-producing genes in siliceous rivers. Due to the differences between benthic growths with respect to planktonic ones, we propose the adoption of the term Cyanobacterial Harmful Algal Mats (CyanoHAMs) as a more precise descriptor for future studies.
Assuntos
Toxinas Bacterianas , Cianobactérias , Tropanos , Microcistinas/análise , Proliferação Nociva de Algas , México , Toxinas Bacterianas/genética , Toxinas Bacterianas/análise , Monitoramento Ambiental , Cianobactérias/genética , Toxinas de Cianobactérias , Rios/microbiologiaRESUMO
The development of simple, reliable, and cost-effective methods is critically important to study the spatial and temporal variation of microcystins (MCs) in the food chain. Nanobodies (Nbs), antigen binding fragments from camelid antibodies, present valuable features for analytical applications. Their small antigen binding site offers a focused recognition of small analytes, reducing spurious cross-reactivity and matrix effects. A high affinity and broad cross-reactivity anti-MCs-Nb, from a llama antibody library, was validated in enzyme linked immunosorbent assay (ELISA), and bound to magnetic particles with an internal standard for pre-concentration in quantitative-matrix-assisted laser desorption ionization-time of flight mass spectrometry (Nb-QMALDI MS). Both methods are easy and fast; ELISA provides a global result, while Nb-QMALDI MS allows for the quantification of individual congeners and showed excellent performance in the fish muscle extracts. The ELISA assay range was 1.8-29 ng/g and for Nb-QMALDI, it was 0.29-29 ng/g fish ww. Fifty-five fish from a MC-containing dam were analyzed by both methods. The correlation ELISA/sum of the MC congeners by Nb-QMALDI-MS was very high (r Spearman = 0.9645, p < 0.0001). Using ROC curves, ELISA cut-off limits were defined to accurately predict the sum of MCs by Nb-QMALDI-MS (100% sensitivity; ≥89% specificity). Both methods were shown to be simple and efficient for screening MCs in fish muscle to prioritize samples for confirmatory methods.
Assuntos
Microcistinas , Anticorpos de Domínio Único , Animais , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Microcistinas/análise , Ensaio de Imunoadsorção EnzimáticaRESUMO
Cyanobacterial blooms are increasingly common in aquatic environments worldwide. These microorganisms cause concern due to their ability to produce cyanotoxins. Aquatic organisms, especially zooplankton, are exposed to cyanobacterial toxins by different routes, depending on the bloom phase. During cyanobacterial dominance, zooplankton is exposed to cyanotoxins through the ingestion of cyanobacterial cells, while at the bloom senescence, dissolved toxins are the most representative route. In this study, we assessed the effects of a microcystin-producing strain of Microcystis aeruginosa (NPLJ-4) on clones of the tropical small cladocerans Macrothrix spinosa (two clones) and Ceriodaphnia cornuta (one clone) exposed to intact cells and aqueous cell crude extracts. Short-term toxicity assays and life-table experiments were performed to assess the effects of the toxic M. aeruginosa on the survival and life history of the cladocerans. In the short-term toxicity assay, we found that cladocerans were more affected by intact cells. Both clones of M. spinosa were more affected when exposed to intact cells, while C. cornuta displayed about 5-fold more resistance. On the other hand, crude extracts had a low impact on cladocerans' survival. Also, we observed a significant decrease in survival, fecundity, and growth of animals exposed to sublethal and environmentally relevant concentrations of M. aeruginosa cellular biomass. However, even at high concentrations of dissolved microcystins, the crude extract did not have significant effects on the life history parameters of the cladocerans. Although they can be found during cyanobacterial bloom events, small-bodied cladocerans are still affected by toxic Cyanobacteria depending on the exposure route.
Assuntos
Cladocera , Cianobactérias , Microcystis , Animais , Extratos Celulares , Ingestão de Alimentos , Microcistinas/toxicidadeRESUMO
Cyanobacteria are photosynthetic microorganisms that compose phytoplankton and therefore have a trophic relationship with zooplankton, which represent an important link for energy flux in aquatic food webs. Several species can form blooms and produce bioactive metabolites known as cyanotoxins. However, the ecological and adaptative role of these toxins are still under debate. Many studies have addressed the cyanotoxins' function in defense against herbivory when grazing pressure by zooplankton plays a role in phytoplankton top-down control. Thus, the present study evaluated the ecophysiological responses of the cyanobacterial strain Microcystis aeruginosa NPLJ-4 underlying the chemical induced defense against the cladoceran Daphnia gessneri. Exposure to predator infochemicals consisted of cultures established in ASM-1 medium prepared in a filtrate from a culture of adults of D. gessneri at an environmentally relevant density. Daphnia infochemicals promoted a significant increase in toxin production by M. aeruginosa. However, no differences in growth were observed, despite a significant increase in both maximum photosynthetic efficiency and electron transport rate in response to zooplankton. Additionally, there was no significant variation in the production of exopolysaccharides. Overall, although a grazer-induced defense response was demonstrated, there were no effects on M. aeruginosa fitness, which maintained its growth in the presence of Daphnia alarm cues.
Assuntos
Daphnia/fisiologia , Cadeia Alimentar , Microcistinas/metabolismo , Microcystis/fisiologia , Feromônios/metabolismo , Animais , Daphnia/química , Comportamento Alimentar , Microcystis/químicaRESUMO
Agriculture runoffs and discharge of wastewaters are the major causes of eutrophication. Although eutrophication could promote the thriving of any phytoplankter, harmful algal blooms (HABs) are dominated frequently by cyanobacteria. Currently, HABs dominated by the toxigenic cyanobacterium Microcystis aeruginosa in lakes and reservoirs are the main environmental concerns worldwide. This study aimed to determine how M. aeruginosa (Ma) modifies the population growth of Pseudokirchneriella subcapitata (Ps) and Ankistrodesmus falcatus (Af). Growth kinetics were determined for each species and in the combinations: Ps-Ma, Af-Ma, Af-Ps, and Ps-Af-Ma. At the end of experiments, photosynthetic pigments, phycobiliproteins, and microcystins were quantified. A logistic equation significantly described the growth trend for all of the tested species, enabling the identification of negative effects on early stages in the population growth of co-cultures with the cyanobacterium; in addition, the interaction effects on the growth rate and in the maximum attainable population density were determined. The biomasses of A. falcatus and P. subcapitata were significantly higher when cultured individually than in all of the combinations with the cyanobacterium. The concentrations of chlorophyll a and b, as well as carotenoids, were lower in combined cultures, but phycobiliprotein content in the cultures with M. aeruginosa was not significantly affected. Microcystis aeruginosa negatively affected the growth of the microalgae, but A. falcatus was significantly more inhibited than P. subcapitata; however, microcystin concentrations were significantly reduced in the co-cultures with microalgae. These results could help to explain the displacements of microalgae when cyanobacteria are present, giving rise to cyanobacterial blooms in eutrophic freshwaters.
Assuntos
Cianobactérias , Microalgas , Microcystis , Clorofila A , Crescimento DemográficoRESUMO
Cyanobacterial toxic blooms are a worldwide problem. The Río de la Plata (RdlP) basin makes up about one fourth of South America areal surface, second only to the Amazonian. Intensive agro-industrial land use and the construction of dams have led to generalized eutrophication of main tributaries and increased the intensity and duration of cyanobacteria blooms. Here we analyse the evolution of an exceptional bloom at the low RdlP basin and Atlantic coast during the summer of 2019. A large array of biological, genetic, meteorological, oceanographic and satellite data is combined to discuss the driving mechanisms. The bloom covered the whole stripe of the RdlP estuary and the Uruguayan Atlantic coasts (around 500 km) for approximately 4 months. It was caused by the Microcystis aeruginosa complex (MAC), which produces hepatotoxins (microcystin). Extreme precipitation in the upstream regions of Uruguay and Negro rivers' basins caused high water flows and discharges. The evolution of meteorological and oceanographic conditions as well as the similarity of organisms' traits in the affected area suggest that the bloom originated in eutrophic reservoirs at the lower RdlP basin, Salto Grande in the Uruguay river, and Negro river reservoirs. High temperatures and weak Eastern winds prompted the rapid dispersion of the bloom over the freshwater plume along the RdlP northern and Atlantic coasts. The long-distance rapid drift allowed active MAC organisms to inoculate freshwater bodies from the Atlantic basin, impacting environments relevant for biodiversity conservation. Climate projections for the RdlP basin suggest an increase in precipitation and river water flux, which, in conjunction with agriculture intensification and dams' construction, might turn this extraordinary event into an ordinary situation.
Assuntos
Cianobactérias , Microcystis , Água Doce , Microcistinas , Rios , América do Sul , UruguaiRESUMO
Cyanobacterial metabolites are increasingly studied, in regards to their biosynthesis, ecological role, toxicity, and potential biomedical applications. However, the history of cyanotoxins prior to the last few decades is virtually unknown. Only a few paleolimnological studies have been undertaken to date, and these have focused exclusively on microcystins and cylindrospermopsins, both successfully identified in lake sediments up to 200 and 4700 years old, respectively. In this paper, we review direct extraction, quantification, and application of cyanotoxins in sediment cores, and put forward future research prospects in this field. Cyanobacterial toxin research is also compared to other paleo-cyanobacteria tools, such as sedimentary pigments, akinetes, and ancient DNA isolation, to identify the role of each tool in reproducing the history of cyanobacteria. Such investigations may also be beneficial for further elucidation of the biological role of cyanotoxins, particularly if coupled with analyses of other abiotic and biotic sedimentary features. In addition, we identify current limitations as well as future directions for applications in the field of paleolimnological studies on cyanotoxins.
Assuntos
Toxinas Bacterianas/toxicidade , Cianobactérias/metabolismo , Limnologia/métodos , Toxinas Marinhas/toxicidade , Microcistinas/toxicidade , Paleontologia/métodos , Toxinas de Cianobactérias , Sedimentos Geológicos , Lagos/microbiologiaRESUMO
Water sources destined to human supply are increasingly threatened worldwide due to various sources of pollution, either point or diffuse. In this sense, the objective of this work was to evaluate the efficiency of an aluminum-based chemical remediator in the Joanes River. An statistical analysis of physical, chemical, biological, and hydraulic monitoring data was performed relying on a 2013-2018 recording period, provided by the local sanitation service provider and the environmental agency. The results showed that even with the use of aluminum-based chemical remediators, the key parameters for controlling flowering events remained high with mean values of 0.18 mg P L-1, 176.155 cells mL-1 of cyanobacteria and peaks of 1.56 µg L-1 and 4.02 µg L-1 for microcystin and saxitoxin, respectively. At the end of this study, it was verified that the aluminum-based chemical remediator showed low effectiveness in the reduction of phosphorus and cyanobacteria, opposing to expectations of the sanitation provider.
Assuntos
Alumínio/toxicidade , Cianobactérias/efeitos dos fármacos , Recuperação e Remediação Ambiental , Poluentes do Solo/toxicidade , Brasil , Monitoramento Ambiental/métodos , Humanos , Microcistinas , Fósforo , Testes de ToxicidadeRESUMO
The cyanobacterium Radiocystis fernandoi has been frequently identified in cyanobacterial blooms in Brazil. Recently, R. fernandoi strain R28, which produces microcystin (MC)-RR and MC-YR, was isolated from the Furnas reservoir, Minas Gerais, Brazil. The present study evaluated the hematological variables and erythrocyte antioxidant responses, lipid peroxidation (LPO), and genotoxicity in a neotropical fish (Hoplias malabaricus) after acute and subchronic exposure to a crude extract (CE) of R. fernandoi strain R28. Acute exposure (12 or 96â¯h) consisted of a single intraperitoneal (i.p.) CE injection, and subchronic exposure consisted of one i.p. CE injection every 72â¯h for 30 days. After acute exposure, fish exhibited macrocytic anemia (12â¯h post-injection) followed by normocytic anemia (96â¯h post-injection). The increased activity of superoxide dismutase, glutathione peroxidase, glutathione-S-transferase, and the glutathione level in the erythrocytes did not prevent oxidative stress, manifested as lipid peroxidation and elevated DNA damage after acute exposure. After subchronic exposure, the hematological variables recovered, and the absence of erythrocyte oxidative stress suggests possible modulation by other biological factors, including a possible decrease in MC uptake by the cells and/or increasing detoxification efficiency that precludes erythrocyte damage.
Assuntos
Misturas Complexas/toxicidade , Cianobactérias/química , Eritrócitos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Animais , Misturas Complexas/química , Misturas Complexas/isolamento & purificação , Dano ao DNA/efeitos dos fármacos , Peixes , Peroxidação de Lipídeos/efeitos dos fármacos , Testes de ToxicidadeRESUMO
Microcystins constitute a serious threat to the quality of drinking water worldwide. However, the eco-physiological role of them is not completely known and it is suggested that toxins can play a role in the antioxidant protection. The objective of this study was to evaluate the microcystin antioxidant capacity in vitro by Electronic Paramagnetic Resonance, highly specific for the different reactive oxygen species and in vivo by 7â¯days exposure of Microcystis aeruginosa to high (29⯰C) temperature in addition to a 26⯰C control condition. An effective in vitro antioxidant activity was observed for [D-Leu1]MC-LR against hydrosoluble radicals. As far as we know, this is the first in vitro record of the role of MC as antioxidant. In addition, a significant increase in cellular biomass was observed under 26⯰C in cultures with [D-Leu1]MC-LR supplementation in coincidence with a significant decrease of reactive species. For cultures at 29⯰C, the antioxidant role of toxins was inconclusive probably due to the presence of different reactive species generated during the experiment. Thus, MC could scavenge certain reactive species associated with the antioxidant role of CAT or the OH content by SOD activity (not measured) and then CAT activity could be lower in the presence of MC. Reinforcing our hypothesis, the [D-Leu1]MC-LR consumption after 7â¯days was significantly higher in cells with [D-Leu1]MC-LR supplementation in both 26⯰C and 29⯰C.When the production of reactive species was controlled by the scavenger activity of antioxidants plus MC, cells avoided the potential oxidative damage and started with exponential growth.
Assuntos
Antioxidantes/fisiologia , Toxinas Bacterianas/metabolismo , Microcistinas/fisiologia , Microcystis , Biomassa , Catalase/metabolismo , Microcystis/crescimento & desenvolvimento , Microcystis/metabolismo , Oxirredução , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo , TemperaturaRESUMO
Absorption and accumulation of bioavailable cyanobacterial metabolites (including cyanotoxins) are likely in fish after senescence and the rupturing of cells during bloom episodes. We determined the toxicity of cyanopeptides identified from two strains of Microcystis (M. panniformis MIRS-04 and M. aeruginosa NPDC-01) in a freshwater tropical fish, Astyanax altiparanae (yellowtail tetra, lambari). Aqueous extracts of both Microcystis strains were prepared in order to simulate realistic fish exposure to these substances in a freshwater environment. Both strains were selected because previous assays evidenced the presence of microcystins (MCs) in MIRS-04 and lack of cyanotoxins in NPDC-01. Identification of cyanobacterial secondary metabolites was performed by LC-HR-QTOF-MS and quantification of the MC-LR was carried out by LC-QqQ-MS/MS. MIRS-04 produces the MCs MC-LR, MC-LY and MC-HilR as well as micropeptins B, 973, 959 and k139. NPCD-01 biosynthetizes microginins FR1, FR2/FR4 and SD-755, but does not produce MCs. Larval fish survival and changes in morphology were assessed for 96 h exposure to aqueous extracts of both strains at environmentally relevant concentrations from 0.1 to 0.5 mg (dry weight)/mL, corresponding to 0.15 to 0.74 µg/mL of MC-LR (considering dried amounts of MIRS-04 for comparison). Fish mortality increased with concentration and time of exposure for both strains of Microcystis. The frequencies of morphological abnormalities increased with concentration in both strains, and included abdominal and pericardial oedema, and spinal curvature. Results demonstrate that toxicity was not solely caused by MCs, other classes of cyanobacterial secondary metabolites contributed to the observed toxicity.
Assuntos
Toxinas Bacterianas/toxicidade , Characidae/anormalidades , Larva/efeitos dos fármacos , Microcystis , Peptídeos/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Larva/crescimento & desenvolvimentoRESUMO
Cyanotoxins from toxic blooms in lakes or eutrophic reservoirs are harmful to several organisms including zooplankton, which often act as vectors of these secondary metabolites, because they consume cyanobacteria, bioaccumulate the cyanotoxins and pass them on along the food chain. Microcystins are among the most commonly found cyanotoxins and often cause zooplankton mortality. Although cyanobacterial blooms are common and persistent in Mexican water bodies, information on the bioaccumulation of cyanotoxins is scarce. In this study we present data on the bioaccumulation of cyanotoxins from Planktothrix agardhii, Microcystis sp., Cylindrospermopsis raciborskii and Dolichospermum planctonicum blooms in the seston (suspended particulate matter more than 1.2⯵m) by zooplankton and fish (tilapia (Oreochromis niloticus) and mesa silverside (Chirostoma jordani) samples from Lake Zumpango (Mexico City). The cyanotoxins were extracted from the seston, zooplankton and fish tissue by disintegration using mechanical homogenization and 75% methanol. After extraction, microcystins were measured using an ELISA kit (Envirologix). Concentration of microcystins expressed as equivalents, reached a maximum value of 117⯵gâ¯g-1 on sestonic samples; in zooplankton they were in the range of 0.0070-0.29⯵gâ¯g-1. The dominant zooplankton taxa included Acanthocyclops americanus copepodites, Daphnia laevis and Bosmina longirostris. Our results indicate twice the permissible limits of microcystins (0.04⯵gâ¯kg-1 d-1) for consumption of cyanobacterial products in whole fish tissue of Chirostoma jordani. The data have been discussed with emphasis on the importance of regular monitoring of water bodies in Mexico to test the ecotoxicological impacts of cyanobacterial blooms and the risk that consumption of products with microcystins could promote.
Assuntos
Monitoramento Ambiental , Peixes/metabolismo , Lagos/microbiologia , Microcistinas/metabolismo , Poluentes Químicos da Água/metabolismo , Zooplâncton/metabolismo , Animais , Copépodes/metabolismo , Cianobactérias/metabolismo , Cylindrospermopsis , Daphnia/metabolismo , Cadeia Alimentar , Lagos/química , México , Microcystis/metabolismo , Alimentos Marinhos/análiseRESUMO
Cyanobacteria can produce potent natural toxins known as cyanotoxins. Blooms of cyanobacteria, produced mainly as result of the pollution of water bodies with excessive amounts of phosphorus, represent a severe environmental problem; not only do they affect the normal equilibrium of the aquatic ecosystem but may also affect animal and human health. The occurrence of algal blooms have been increasing globally (it has been recently reported in at least 100 countries) and it has been considered by WHO as an emerging public health issue. The toxic effects of cyanotoxins have been thoroughly demonstrated in laboratory experiments, however, the effects on humans and the extent of these effects have been more difficult to assess. Epidemiological research is difficult as there are no specific symptoms or routine biomarkers to diagnose intoxication with cyanotoxins, in particular those cases associated with chronic exposure. The objectives of this study were to assess the exposure of a population settled near a lake with recurrent cyanobacteria blooms and to investigate the presence of biological markers of chronic exposure to cyanotoxins, in particular the microcystins (MCs). We first investigated the exposure of the population to cyanobacteria by using a questionnaire on how the population used the water and by analyzing water samples for the presence of cyanobacteria and total microcystins (TMCs). Secondly, we investigated the presence of biological indicators by analyzing the biochemical and immunological parameters in sera of the exposed population. The questionnaires and the water analyses revealed that the population under study (nâ¯=â¯47) is exposed to several exposure routes. The biochemical analyses of the sera showed the alteration of at least one hepatic enzyme in 25% of the exposed people, but this cannot be associated solely to MCs exposure. On the contrary, the immunological analyses, which included microcystin-LR specific antibodies IgE and IgG, showed significant differences between the exposed and non-exposed groups. The presence of MCs specific antibodies confirms the exposure to MCs. We propose the study of specific antibodies as a non-complex biomarker to detect chronic exposure to the toxin and to assist epidemiological studies.
Assuntos
Cianobactérias , Biomarcadores Ambientais/efeitos dos fármacos , Exposição Ambiental/análise , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Microcistinas/toxicidade , Poluentes Químicos da Água/toxicidade , Argentina , Cianobactérias/crescimento & desenvolvimento , Ecossistema , Eutrofização , Humanos , Lagos/química , Toxinas Marinhas , Inquéritos e QuestionáriosRESUMO
The presence of cyanobacterial blooms and cyanotoxins in water presents a global problem due to the deterioration of ecosystems and the possibility of poisoning in human and animals. Microcystin LR is the most widely distributed cyanotoxin and liver cells are its main target. In the present study, HepG2 cells were used to determine DNA damage of three crude extracts of cyanobacterial blooms containing MC-LR, through comet assay. The results show that all extracts at a concentration of 500⯵gâ¯mL-1 caused low damage in hepatocytes exposed for 24â¯h, but produced total mortality even at low concentrations at 48â¯h. Moreover, balloons corresponding to cell apoptosis were found. Through HPLC/MS, MC-LR was detected in all samples of cyanobacterial blooms at concentrations of (5,65⯵gâ¯ml-1) in sample 1, (1,24⯵gâ¯ml-1) in sample 2 and (57,29⯵gâ¯ml-1) in sample 3. In addition, in all samples high molecular weights peaks were detected, that may correspond to other microcystins. Besides, the cytotoxic effect of a cyanobacterial bloom and some of its chromatographic fractions from the crude extracts were evaluated in U-937, J774, Hela and Vero cell lines, using the enzymatic micromethod (MTT). The highest toxicity was detected in U-937â¯cells (LC50â¯=â¯29.7⯵gâ¯mL-1) and Vero cells (LC50â¯=â¯39.7⯵g mL-1). Based on these results, it is important to remark that genotoxic and cytotoxicity assays are valuable methods to predict potential biological risks in waters contaminated with blooms of cyanobacteria, since chemical analysis can only describe the presence of cyanotoxins, but not their biological effects.
Assuntos
Cianobactérias/química , Microcistinas/toxicidade , Animais , Compostos de Benzil , Células Cultivadas , Chlorocebus aethiops , Colômbia , Ensaio Cometa , Cianobactérias/metabolismo , Dano ao DNA , Monitoramento Ambiental , Células Hep G2 , Humanos , Toxinas Marinhas , Microcistinas/análise , Microcistinas/química , Microcistinas/metabolismo , Testes de Mutagenicidade/métodos , Pirazinas , Células U937 , Células VeroRESUMO
The occurrence of harmful algal blooms in nutrient-rich freshwater bodies has increased world-wide, including in the Pacific Northwest. Some cyanobacterial genera have the potential to produce secondary metabolites that are highly toxic to humans, livestock and wildlife. Reliable methods for the detection of cyanobacterial toxins with high specificity and low limits of detection are in high demand. Here we test a relatively new hybrid high resolution accurate mass quadrupole time-of-flight mass spectrometry platform (TripleTOF) for the analysis of cyanobacterial toxins in freshwater samples. We developed a new method that allows the quantitative analysis of four commonly observed microcystin congeners (LR, LA, YR, and RR) and anatoxin-a in a 6-min LC run without solid-phase enrichment. Limits of detection for the microcystin congeners (LR, LA, YR, and RR) and anatoxin-a were <5 ng/L (200-fold lower than the guideline value of 1 µg/L as maximum allowable concentration of MC-LR in drinking water). The method was applied for screening freshwaters in the Pacific Northwest during the bloom and post-bloom periods. The use of high resolution mass spectrometry and concomitant high sensitivity detection of specific fragment ions with high mass accuracy provides an integrated approach for the simultaneous identification and quantification of cyanobacterial toxins. The method is sensitive enough for detecting the toxins in single Microcystis colonies.
RESUMO
Cyanobacterial proliferation in river and lakes is the result of eutrophication. The cyanobacterium Radiocystis fernandoi strain R28 produces mostly two MC variants MC-RR and MC-YR and small amounts of other oligopeptides, but does not produce MC-LR. The present study investigated the hepatotoxic potential of the crude extract of the R. fernandoi strain R28 on the Neotropical fish, Piaractus mesopotamicus, at 3, 6, and 24 h after intraperitoneal injection (100 µg MC-LR equivalent per kg-1 body mass) using biochemical and morphological biomarkers of liver damage. Although the protein phosphatases PP1 and PP2A were not inhibited during the 24-h treatment, liver parenchyma and hepatocyte structure were disrupted. Alkaline phosphatase increased at 3 h post-injection and decreased after 24 h; alanine aminotransferase and aspartate aminotransferase increased in a time-dependent manner up to 24 h indicating impaired liver function. Progressive histopathological changes were consistent with biochemical results demonstrating alterations in liver structure and function. In conclusion, the crude extract of R. fernandoi strain R28 has high hepatotoxic potential and can severely compromise fish health.
Assuntos
Alanina Transaminase/metabolismo , Fosfatase Alcalina/metabolismo , Aspartato Aminotransferases/metabolismo , Biomarcadores/metabolismo , Cianobactérias/efeitos dos fármacos , Hepatócitos/efeitos dos fármacos , Microcistinas/metabolismo , Alanina Transaminase/química , Fosfatase Alcalina/química , Animais , Aspartato Aminotransferases/química , Peixes , Injeções Intraperitoneais , Lagos , HepatopatiasRESUMO
Microcystins (MCs) are potent hepatotoxins, and their presence in water bodies poses a threat to wildlife and human populations. Most of the available information refers to plankton, and much less is known about microcystins in other habitats. To broaden our understanding of the presence and environmental distribution of this group of toxins, we conducted extensive sampling throughout Spain, under a range of conditions and in distinct aquatic and terrestrial habitats. More than half of the tested strains were toxic; concentrations of the hepatotoxin were low compared with planktic communities, and the number of toxic variants identified in each sample of the Spanish strains ranged from 1-3. The presence of microcystins LF and LY (MC-LF and MC-LY) in the tested samples was significant, and ranged from 21.4% to 100% of the total microcystins per strain. These strains were only detected in cyanobacteria Oscillatoriales and Nostocales. We can report, for the first time, seven new species of microcystin producers in high mountain rivers and chasmoendolithic communities. This is the first report of these species in Geitlerinema and the confirmation of Anatoxin-a in Phormidium uncinatum. Our findings show that microcystins are widespread in all habitat types, including both aerophytic and endolithic peat bogs and that it is necessary to identify all the variants of microcystins in aquatic bodies as the commonest toxins sometimes represent a very low proportion of the total.
Assuntos
Cianobactérias/isolamento & purificação , Microcistinas/análise , Tropanos/análise , Poluentes da Água/análise , Toxinas de Cianobactérias , Ecossistema , Monitoramento Ambiental , Nascentes Naturais/microbiologia , Rios/microbiologia , EspanhaRESUMO
BACKGROUND: Acute exposure to high concentrations of microcystin-LR (MC-LR) can cause significant hepatocyte injury. AIM: To document the effects of long-term, low-dose MC-LR exposure on hepatic inflammation and fibrosis in mice with healthy and diseased livers. MATERIAL AND METHODS: Male CD1 mice (N = 20/group) were exposed to 1.0 µg/L of MC-LR in drinking water; 1.0 µg/L MC-LR plus 300 mg/L of the hepatotoxin thioacetamide (MC-LR/TAA); or 300 mg/L TAA alone for 28 weeks. Liver biochemistry and histology were documented at the end of the study period. In addition, hepatic stellate cells (HSCs), were exposed in vitro to MC-LR (0.1-10,000 µg/L) and monitored for changes in cell metabolism, proliferation and activation. RESULTS: Liver biochemistry and histology were essentially normal in MC-LR alone exposed mice. MC-LR/TAA and TAA alone exposed mice had significant hepatic inflammation and fibrosis but the extent of the changes were similar in the two groups. In vitro, MC-LR had no effect on HSC metabolism, proliferation or activation. CONCLUSION: Long-term, low-dose exposure to MC-LR is unlikely to lead to chronic liver disease in the setting of a normal liver or exacerbate existing liver disease in the setting of ongoing hepatitis.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas/etiologia , Células Estreladas do Fígado/efeitos dos fármacos , Cirrose Hepática Experimental/induzido quimicamente , Fígado/efeitos dos fármacos , Microcistinas/toxicidade , Animais , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/patologia , Metabolismo Energético/efeitos dos fármacos , Células Estreladas do Fígado/metabolismo , Células Estreladas do Fígado/patologia , Fígado/metabolismo , Fígado/patologia , Cirrose Hepática Experimental/metabolismo , Cirrose Hepática Experimental/patologia , Masculino , Toxinas Marinhas , Camundongos , Medição de Risco , Índice de Gravidade de Doença , Tioacetamida , Fatores de TempoRESUMO
In January 2015, a 20-month-old child and her family took part in recreational activities at Carrasco and Malvín beaches (Montevideo, Uruguay). An intense harmful algae bloom (HAB) was developing along the coast at that time. A few hours after the last recreational exposure episode, the family suffered gastrointestinal symptoms which were self-limited except in the child's case, who was admitted to hospital in Uruguay with diarrhea, vomiting, fatigue, and jaundice. The patient had increased serum levels of liver enzymes and bilirubin and five days later presented acute liver failure. She was referred to the Italian Hospital in Buenos Aires, being admitted with grade II-III encephalopathy and hepatomegaly and requiring mechanical respiratory assistance. Serology tests for hepatitis A, B, and C, Epstein-Barr virus, and cytomegalovirus were negative. Laboratory features showed anemia, coagulopathy, and increased serum levels of ammonium, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and bilirubin. Autoimmune Hepatitis Type-II (AH-II) was the initial diagnosis based on a liver kidney microsomal type 1 antibodies (LKM-1) positive result, and twenty days later a liver transplant was performed. The liver histopathology had indicated hemorrhagic necrosis in zone 3, and cholestasis and nodular regeneration, which were not characteristic of AH-II. LC/ESI-HRMS (liquid chromatography electrospray ionization high-resolution mass spectrometry) analysis of MCs in the explanted liver revealed the presence of Microsytin-LR (MC-LR) (2.4 ng·gr-1 tissue) and [D-Leu¹]MC-LR (75.4 ng·gr-1 tissue), which constitute a toxicological nexus and indicate a preponderant role of microcystins in the development of fulminant hepatitis.
Assuntos
Proliferação Nociva de Algas , Falência Hepática/etiologia , Microcistinas/toxicidade , Poluentes da Água/toxicidade , Alanina Transaminase/sangue , Aspartato Aminotransferases/sangue , Praias , Bilirrubina/sangue , Exposição Ambiental , Feminino , Humanos , Lactente , Fígado/metabolismo , Fígado/patologia , Falência Hepática/sangue , Falência Hepática/patologia , UruguaiRESUMO
Cyanobacterial blooms are becoming increasingly common in aquatic environments around the world, mainly due to eutrophication and climate change. Cyanotoxin-producing strains (e.g., microcystins (MC) producers) may be present in these blooms, affecting the growth of other aquatic organisms, such as aquatic macrophytes. In this study, we evaluated the morphometric and physiological responses of the aquatic macrophyte Egeria densa to the exposure to a toxic strain of Microcystis aeruginosa (MCs producer) and a non-toxic Microcystis panniformis (non-MC producer). The effects of Microcystis strains on E. densa growth and biomass were verified for five weeks (Experiment 1) and physiological responses were evaluated for 14days (Experiment 2). Prolonged exposure of E. densa to the MC producing strain reduced growth, accompanied by the inhibition of shoot and root emission. Both Microcystis strains caused a decrease in the content of photosynthetic pigments, like total chlorophyll and chlorophyll a and b, accompanied by an increase of carotenoids. At the beginning of the MC-producing strain exposure, E. densa showed an increase in the activity of the anti-oxidative enzymes superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX), accompanied by an increase in the levels of malondialdehyde, indicating lipid peroxidation. During the 14th day of exposure, the activity of antioxidant enzymes remained similar to the control, suggesting that E. densa has an efficient anti-oxidative system to control the reactive oxygen species produced in response to the stress caused by microcystins. However, when prolonged exposure occurred, possible damage to proteins may have affected the growth and development of E. densa. No changes were observed in the enzymatic activity of the plants exposed to the non-MC producing strain, suggesting that this cyanobacterial strain do not cause significant damage to the development of E. densa. These results are important for understanding the anti-oxidative defense mechanisms of aquatic macrophytes when coexisting with an MC producing strain.