Distribución espacio-temporal de cianobacterias planctónicas y factores ambientales asociados a sus proliferaciones en el embalse Cerrón Grande, El Salvador / Spatio-temporal distribution of planktonic cyanobacteria and environmental factors associated with their proliferation in the Cerrón Grande reservoir, El Salvador

Resumen Introducción: El embalse Cerrón Grande de 135 km2 es el humedal más grande de El Salvador, reconocido como "Humedal de Importancia Internacional" por la Convención Ramsar. Además, es uno de los ecosistemas más contaminados a nivel nacional, enfrentando serios problemas, entre ellos, las proliferaciones de cianobacterias. Objetivo: Evaluar las relaciones entre los factores ambientales y la distribución espacial y temporal de las cianobacterias en el embalse a lo largo de un año hidrológico. Métodos: Extrajimos agua con cianobacterias de seis sitios fijos, desde octubre 2018 hasta septiembre 2019. Resultados: Identificamos 16 géneros, el más abundante Microcystis sp. Además, Dolichospermum sp.; Microcistis sp. alcanzaron 1.5 x 106 células/ml en junio 2019, principalmente en el sector noroeste, que es poco profundo y cercano a las zonas de confluencia de afluentes altamente contaminados que llegan al embalse. El sector sureste más profundo tenía bajas concentraciones de estos organismos. Los factores ambientales mayormente relacionados con dicho comportamiento fueron: fósforo, la conductividad eléctrica y el valor de Secchi, como se esperaba de un cuerpo eutrófico. Conclusiones: La variación en la concentración celular de cianobacterias en este embalse está altamente influenciada por la lluvia, la mezcla de agua y el contenido de nutrientes.

Abstract Introduction: The 135 km2 Cerrón Grande reservoir is the largest wetland in El Salvador, recognized as "Wetland of International Importance" by the Ramsar Convention. It is also one of the most polluted ecosystems nationwide, facing serious problems, among them, a proliferation of cyanobacteria. Objective: To assess the relationships among environmental factors and the spatial and temporary distribution of cyanobacteria in the reservoir throughout a hydrological year. Methods: We extracted water with cyanobacteria from six fixed sites, from October 2018 through September 2019. Results: We identified 16 genera, the most abundant Microcystis sp. And Dolichospermum sp.; Microcystis sp. reached 1.5 x 106 cells/ml in June 2019, mainly in the north-west sector, which is shallow and close to the confluence zones where highly polluted tributaries reach the reservoir. The deeper south-east sector had low concentrations of these organisms. The environmental factors mostly related to this behavior were: phosphorus, electrical conductivity and Secchi value, as expected from a eutrophic body. Conclusions: Variation in the cellular concentration of cyanobacteria in this reservoir is highly influenced by rain, water mixing and nutrient content.

Effects of homeopathic preparations on raphidiopsis raciborskii (Cyanobacteria) toxicity in artemia salina mode

Cyanobacteria are microorganisms found in different parts of the world. Some genera are cyanotoxins producers a sodium channel blockingneurotoxin (saxitoxins). Some homeopathic preparations have been identified as remedial action on toxicity models in Artemia salina. This study aimed to observe whether homeopathic products influence the toxicity ofR.raciborskiiextract onA.salinaby inducing cyst hatching arrest, anembryo bioresiliencemodel previously developed in our laboratory (Pinto et al., 2021; Mohammad et al., 2022). Thus, previous toxicity testswere carried out on cysts in 96-well plates, using different concentrations of the extract obtained from regular cultivation of R. raciborskii in HCl 0.05M, whose strain, named T3, is kept in the laboratory of Cyanobacteria at FURGS, Brazil.The standardization of toxin concentration was based on an established scale developed at FURGS, in which the number of T3 filaments is associated withspecific saxitoxin concentrationsdefined by chromatography. The concentration of 2.6 µg/L was chosen since it reducedthe cysthatching rate by 30%, the ideal level to observe embryo bioresilience. Then, a screeningstudy with 22 homeopathic preparations was tested blind in three experimental series, in duplicate,against threecontrols (unchallenged, water,and succussed water)for possible toxicity attenuationon Artemia salinacysts hatching rate. Homeopathic medicines were prepared in pure,sterile water from a stock homeopathic solution, one potency below the working potency. After the 1:100 dilution, 100 succussions were made using a robotic arm (Denise, Autic). The medicines were inserted into the seawater on a 10% basis. Due to the high sensitivity of A. salinato the circalunar variations, all experiments were performed during the first quarter moon. Statistical analysis was performed by two-way ANOVA followed by Tukey, with α=0.05. The most significant results indicative of bioresilience improvement were seen after the treatment with Nitric acidum6 cH, Plumbum metallicum6 cH, isotherapic 200 cH, and hydrochloric acid 1 cH being the last one used as a vehicle of the extracts. Thus, these preparations were chosen to be used in further experiments. In conclusion, the Artemia salinamodel has also beenuseful to study bioresilienceimprovement by homeopathic medicines after intoxication with saxitoxin.

A new method for isolating gas vesicles from Microcystis for ultrasound contrast / 生物工程学报

Gas vesicles are a unique class of gas-filled protein nanostructures which are commonly found in cyanobacteria and Halobacterium. The gas vesicles may scatter sound waves and generate harmonic signals, which enabled them to have the potential to become a novel ultrasound contrast agent. However, the current hypertonic cracking method for isolating gas vesicles contains tedious operational procedures and is of low yield, thus not suitable for large-scale application. To overcome these technical challenges, we developed a rapid and efficient method for isolating gas vesicles from Microcystis. The new H2O2-based method increased the yield by three times and shortened the operation time from 24 hours to 7 hours. The H2O2 method is not only suitable for isolation of gas vesicles from laboratory-cultured Microcystis, but also suitable for colonial Microcystis covered with gelatinous sheath. The gas vesicles isolated by H2O2 method showed good performance in ultrasound contrast imaging. In conclusion, this new method shows great potential for large-scale application due to its high efficiency and wide adaptability, and provides technical support for developing gas vesicles into a biosynthetic ultrasonic contrast agent.

Engineering the glycogen metabolism in cyanobacterial photosynthetic cell factories: a review / 生物工程学报

Cyanobacteria are important photosynthetic autotrophic microorganisms and are considered as one of the most promising microbial chassises for photosynthetic cell factories. Glycogen is the most important natural carbon sink of cyanobacteria, playing important roles in regulating its intracellular carbon distributions. In order to optimize the performances of cyanobacterial photosynthetic cell factories and drive more photosynthetic carbon flow toward the synthesis of desired metabolites, many strategies and approaches have been developed to manipulate the glycogen metabolism in cyanobacteria. However, the disturbances on glycogen metabolism usually cause complex effects on the physiology and metabolism of cyanobacterial cells. Moreover, the effects on synthesis efficiencies of different photosynthetic cell factories usually differ. In this manuscript, we summarized the recent progress on engineering cyanobacterial glycogen metabolism, analyzed and compared the physiological and metabolism effects caused by engineering glycogen metabolism in different cyanobacteria species, and prospected the future trends of this strategy on optimizing cyanobacterial photosynthetic cell factories.

Growth of Microcystisstrains isolated from environments with the presence and absence of submerged macrophytes in coexistence with Ceratophyllum demersum

Cyanobacterial blooms can cause severe ecological and health problems in drinking water reservoirs. To alleviate this problem, allelopathically active submerged macrophytes can be used to reduce cyanobacterial growth. Accordingly, this study aimed to evaluate the sensitivity of strains of the Microcystis aeruginosacomplex isolated from reservoirs with the presence and absence of submerged macrophytes to the allelochemicals of Ceratophyllum demersum.A coexistence experiment was carried out between the submerged macrophyte C. demersum and four Microcystisstrains, with two treatments for each strain, one in coexistence with the submerged macrophyte (7 g L-1) and control (in the absence of the macrophyte). Two strains of M. aeruginosa(BMIUFRPE-06 and BMIUFRPE-07) and two of M. panniformis(BMIUFRPE-08 and BMIUFRPE-09) were used, which were isolated from Cajueiro (with submerged macrophytes) and Tapacurá (without submerged macrophytes) reservoirs, respectively. The biomass of Microcystisstrains from the reservoir without macrophytes (BMIUFRPE-08 and BMIUFRPE-09) was significantly inhibited in 96% (T-test: p 0.05; growth rate -ANOVA: p > 0.05). These results suggest that strains isolated from environments with submerged macrophytes are less sensitive to allelochemicals of these plants,as these strains may be adapted to the coexistence with submerged macrophytes.

Métodos de extração, estabilidade e biodisponibilidade da guanitoxina em cultivos de Sphaeropermopsis torques-reginae analisadas por espectrometria de massas / Methods of extraction, stability, and bioavailability of guanitoxin in Sphaeropermopsis torques-reginae cultures analyzed by mass spectrometry

A guanitoxina (GNT) é uma neurotoxina produzida por algumas cepas de cianobactérias dos gêneros Dolichospermum e Sphaerospermopsis>. A GNT é o único organofosforado natural, capaz de causar a morte de animais selvagens e domésticos devido à inibição irreversível da acetilcolinesterase. Apesar de sua alta toxicidade, o diagnóstico da GNT em amostras biológicas ainda é um grande desafio. A dificuldade para sua detecção está diretamente ligada à sua instabilidade em altas temperaturas e pH alcalino, tornando difícil seu monitoramento em corpos d'água. Por isso, esta pesquisa objetivou estudar a estabilidade e biodisponibilidade da GNT em amostras aquosas, com intuito de obter mais informações sobre a natureza química e biológica dessa potente neurotoxina. Para realizar este estudo, a cepa ITEP-24 (S. torques-reginae) produtora de GNT foi cultivada em laboratório sob condições controladas, para obter biomassa para os experimentos de extração, semi-isolamento, estabilidade, ensaio in vitro e identificação por LC-MS/MS. Primeiramente foram realizados testes de extração da GNT partir de células liofilizadas da cepa ITEP-24 utilizando água, metanol e etanol em pH ácido. Depois utilizou-se dois métodos de extração em fase sólida (SPE) com cartuchos preenchidos com fases estacionarias C18 em fase reversa e sílica gel em fase normal, com objetivo de avaliar qual método de SPE seria melhor para extrair e concentrar a GNT. Nós também testamos métodos para lisar as células com sondas de ultrassom, misturador e centrifugação. Além dos métodos de extração, nós avaliamos a estabilidade da toxina em diferentes temperaturas, para isso a biomassa seca contendo a GNT ficou condicionada a 4 °C, 23 °C, -20 °C, -80 °C durantes seis meses, e análises de identificação foram realizadas dentro período de 150 dias em uma sequência de 30 dias. A estabilidade da toxina foi analisada também a partir de extrações em soluções com diferentes valores de pH (1,5; 3,0; 5,0; 7,0; 8,5; 10,5) e temperatura (23 ºC e 37 ºC). Depois, analisou-se a biodisponibilidade da GNT em células frescas da linhagem ITEP-24 através de teste de dissolução in vitro. O objetivo deste teste foi avaliar a liberação da toxina intracelular em meio simulado do conteúdo gástrica e intestinal com e sem enzimas digestivas para compreender e estimar a disponibilidade da GNT in vivo. Os resultados de todos experimentos descritos neste estudo, foram obtidos a partir de análises por cromatografia líquida de interação hidrofílica (HILIC) acoplado ao espectrômetro de massas do tipo triplo quadrupolo LC-QqQ-MS/MS utilizando as transições 253>58, 253>159 e 159>58 [M+H]+ utilizando coluna com fase estacionária zwitteriônica (ZIC). A identificação da GNT foi realizada também por cromatografia líquida acoplada ao espectrômetro de massas de alta resolução (LC-HR-QTOF-MS) com coluna Luna C18, Hydro-RP C18 e ZIC-HILIC. Dos protocolos de extração testados, a combinação de metanol/água (70:30 v/v) com ácido acético (0.3%) extraiu maior quantidade relativa da GNT a partir de células frescas e liofilizadas da cepa ITEP-24 e a concentração da toxina foi maior em amostras de células frescas. Em relação aos métodos de lise celular, as extrações realizadas em sonda de ultrassom com banho-maria e centrifugação por 1h foram estatisticamente significantes para liberar a toxina intracelular. Não houve diferença significativa entre os testes de SPE, no entanto, a semipurificação da toxina foi melhor com cartucho preenchido com sílica gel em fase normal e adaptação desse método em coluna aberta permitiu obter uma fração enriquecida com GNT. A GNT mostrou ser mais estável em pH ácido, sendo o pH 3,0 o melhor para manter e extrair a toxina em amostras aquosas e a toxina intracelular presente em células secas podem degradar em temperatura de 23 °C por um período de 150 dias mesmo em solução com pH 3,0. Durante os testes de extração e purificação foi observado também a degradação da toxina em processos de secagem e ressuspensão. As análises realizadas no LC-HR-QTOF-MS com diferentes métodos cromatográficos possibilitou a identificação da GNT, porém o método realizado com coluna ZIC-HILIC mostrou melhor resolução cromatográfica dos picos relativos m/z e tempo de retenção de toxina. Os resultados obtidos nos testes de dissolução in vitro mostraram que a GNT fica mais disponível no simulado gástrico com e sem a enzima pepsina, mas também pode ser absorvida no intestino. Portanto, o teste de dissolução in vitro pode ser uma ferramenta útil para a avaliação de risco de cianotoxinas in vivo, devido ao seu potencial de monitorar qualitativa e quantitativamente substâncias dissolvidas em fluidos gastrointestinais. Os resultados apresentados neste estudo fornecem informações valiosas para uma melhor compreensão da estabilidade e biodisponibilidade do GNT. Além disso, os métodos apresentados neste estudo podem ser úteis para diversas aplicações projetadas para identificar a toxina em amostras ambientais, bem como orientações para procedimentos de purificação da GNT

Guanitoxin (GNT) is a neurotoxin produced by some strains of cyanobacteria of the genus Dolichospermum and Sphaerospermopsis. GNT is the only natural organophosphate, capable of causing the death of animals from wild and domestic animals due to irreversible inhibition of acetylcholinesterase. Despite its high toxicity, the diagnosis of GNT in biological samples is still a significant challenge. The difficulty in its detection is directly linked to its instability at high temperatures and alkaline pH, making it difficult to monitor in bodies of water. Therefore, this research aimed to study the stability and bioavailability of GNT in aqueous samples to provide more information about the chemical and biological nature of this molecule. The strain ITEP-24 (S. torques-reginae) producing GNT was grown in the laboratory under controlled conditions to obtain biomass for the extraction, semi-isolation, stability, in vitro tests, and toxin identification by LC-MS/MS. Firstly, tests were carried out to extract GNT from lyophilized cells strain ITEP-24 using water, methanol, and ethanol at acidic pH and, two SPE methods in cartridges with stationary phases of C18 reverse phase and normal phase gel silica, to evaluate which would be better to extract and concentrate the GNT. We also tested different methods of cell lysis, such as ultrasound probes, mixers, and centrifugation. In addition to the extraction methods, the stability of the toxin was evaluated at different temperatures, for this, the dry biomass containing the toxin was conditioned at 4 °C, 23 °C, -20 °C, -80 °C for 150 days and analysis of the identification of the GNT was carried out within that period in a sequence of 30 days. The toxin stability was also analyzed from extractions in solutions with different pH values (1.5; 3.0; 5.0; 7.0; 8.5; 10.5) and temperature (23 ºC and 37 ºC). In addition, we performed dissolution tests with fresh cells of the ITEP-24 strain to evaluate the bioavailability of GNT in simulated gastric and intestinal fluids with and without digestive enzymes to understand and estimate the availability of GNT in vivo. The results of all experiments described in this study were obtained from analyzes by hydrophilic interaction liquid chromatography (HILIC) coupled to the LC-QqQ-MS/MS triple quadrupole mass spectrometer using the transitions m/z 253> 58, m/z 253> 159 and m/z 159> 58 [M + H]+ using a column with the zwitterionic stationary phase (ZIC). Liquid chromatography coupled to the high-resolution mass spectrometer (LC-HR-QTOF-MS) with Luna column C18, Hydro-RP C18, and ZIC-HILIC carried out the identification of the GNT. From the extraction protocols tested, the combination of methanol/water (70:30 v/v) with acetic acid (0.3%) extracted a greater relative amount of GNT from fresh and lyophilized ITEP-24 cells, and the concentration of the toxin is higher previously fresh. Concerning cellular methods, the ultrasound probe with a water bath and centrifugation for 1h ware statistically significant to release the intracellular toxin. There was no significant difference between the SPE tests. However, the semi-purification of the toxin was better with a cartridge filled with gel silica in the normal phase and adaptation of this method in an open column allowed to obtain a fraction enriched with GNT. GNT was more stable at acid pH, with pH 3.0 being the best to maintain and the intracellular toxin present in dry cells can degrade at a temperature at 23 °C for 150 days even in pH 3.0 solution. The toxin can also hydrolyze in the drying and resuspension processes. The analyzes carried out in LC-HR-QTOF-MS with different chromatographic methods made it possible to identify the GNT itself, however, the ZIC-HILIC column method showed excellent chromatographic resolution of the relative m/z peaks and toxin retention time. The results obtained in the in vitro dissolution tests showed that GNT is more available in the gastric simulation with and without the enzyme pepsin, but it can also be absorbed in the intestine. Thus, in vitro dissolution tests can be used as a useful tool for the risk assessment of cyanotoxins in vivo due to their potential to qualitatively and quantitatively monitor substances dissolved in gastrointestinal fluids. The results presented in this study provide valuable information for a better understanding of the stability and bioavailability of GNT. Besides, the methods presented in this study can be useful for various applications designed to identify the toxin in environmental samples, as well as guidance on procedures for purifying GNT

Potential Biotechnological Applications of Cyanobacterial Exopolysaccharides

Abstract The cyanobacterial exopolysaccharides (EPSs) are considered as one of the important group of biopolymers having significant ecological, industrial, and biotechnological importance. Cyanobacteria are regarded as a very abundant source of structurally diverse, high molecular weight polysaccharides having variable composition and roles according to the organisms and the environmental conditions in which they are produced. Due to their structural complexity, versatility and valuable biological properties, they are now emerging as high-value compounds. They are possessing exceptional properties and thus are being widely explored for various applications like in food and pharmaceutical industries, in bioremediation for removal of heavy metals, for soil conditioning, as biopolymers, bioadhesives, and bioflocculants. However, poor understanding of their complex structural properties, lack of concrete information regarding the genes encoding the proteins involved in the EPS biosynthetic pathways, their process of production and about the associated factors controlling their structural stability, strongly limits their commercialization and applications in the various fields of biotechnology. Owing to the above context, the present review is aimed to organize the available information on applications of cyanobacterial EPSs in the field of biotechnology and to identify the research gaps for improved industrial utilization and commercialization of these biomaterials.

Production of lactate from carbon fixation by cyanobacteria: development and prospect / 生物工程学报

Lactate is an important industrial chemical and widely used in various industries. In recent years, with the increasing demand for polylactic acid (PLA), the demand for lactate raw materials is also increasing. The contradiction between the high cost and the market demand caused by the heterotrophic production of lactate attracts researchers to seek other favorable solutions. The production of lactate from photosynthetic carbon fixation by cyanobacteria is a potential new raw material supply strategy. Based on the photosynthetic autotrophic cell factory, it can directly produce high optical purity lactate from carbon dioxide on a single platform driven by solar energy. The raw materials are cheap and easy to obtain, the process is simple and controllable, the products are clear and easy to separate, and the double effects of energy saving and emission reduction and production of high value-added products are achieved at the same time, which has important research and application value. This paper reviews the development history of cyanobacteria carbon sequestration to produce lactate, summarizes its research progress and encounters technical difficulties from the aspects of metabolic basis, metabolic engineering strategy, metabolic kinetics analysis and technical application, and prospects the future of this technology.

Advances of polymer-monomer production by cyanobacterial cell factory / 生物工程学报

Cyanobacteria is one of the promising microbial chassis in synthetic biology, which serves as a typical host for light-driven production. With the gradual depletion of fossil resources and intensification of global warming, the research on cyanobacterial cell factory using CO2 as carbon resource is ushering in a new wave. For a long time, research focus on cyanobacterial cell factory has mainly been the production of energy products, such as liquid fuels and hydrogen. One of the critical bottlenecks occurring in cyanobacterial cell factory is the poor economic performance, which is mainly caused by the inherent inefficiency of cyanobacteria. The problem is particularly prominent for these extremely cost-sensitive energy products. As an indispensable basis for modern industry, polymer monomers belong to the bulk chemicals with high added value. Therefore, increasing attention has been focused on polymer monomers which are superior in overcoming the economic barrier in commercialization of cyanobacterial cell factories. Here, we systematically review the progress on the production of polymer monomers using cyanobacteria, including the strategies for improving production, and the related technologies for the application of this important microbial cell factory. Finally, we summarize several issues in cyanobacterial synthetic biology and proposed future developing trends in this field.

Heterologous expression and function evaluation of Gloeobacter violaceus rhodopsin in Escherichia coli / 生物工程学报

Proton-pumping rhodopsin (PPR) is a simple photosystem widely distributed in nature. By binding to retinal, PPR can transfer protons from the cytoplasmic to the extracellular side of the membrane under illumination, creating a proton motive force (PMF) to synthesize ATP. The conversion of light into chemical energy by introducing rhodopsin into nonphotosynthetic engineered strains could contribute to promoting growth, increasing production and improving cell tolerance of microbial hosts. Gloeorhodopsin (GR) is a PPR from Gloeobacter violaceus PCC 7421. We expressed GR heterologously in Escherichia coli and verified its functional activity. GR could properly function as a light-driven proton pump and its absorption maximum was at 539 nm. We observed that GR was mainly located on the cell membrane and no inclusion body could be found. After increasing expression level by ribosome binding site optimization, intracellular ATP increased, suggesting that GR could supply additional energy to heterologous hosts under given conditions.

Crecimiento y metabolismo de Fischerella TB22 en medio de cultivo BG11 / Growth and metabolism of Fischerella TB22 in BG110 culture medium

Los microorganismos fijadores de nitrógeno de vida libre, abarcan una gama morfológica que va desde los organismos unicelulares como las bacterias y algunas cianobacterias, hasta multicelulares, filamentosas, por ello es importante conocer cómo se comportan y se puede saber haciendo una curva de crecimiento microbiano. Para este estudio se prepararon 4 fotobioreactores de columna burbujeada con inoculo de Fischerella TB22, se pusieron en aireación constante con 12 horas luz y 12 horas obscuridad durante 40 días con diferentes tratamientos de ajuste de volumen del medio de cultivo y ajuste del pH. El objetivo de este trabajo fue evaluar el crecimiento en biomasa por peso seco, densidad óptica, pH y amonio de Fischerella sp. en medio de cultivo BG110 durante 12 días. Las variables que se midieron de la curva de crecimiento de las cianobacterias, siguieron el patrón de una curva típica de crecimiento microbiano. (AU)

Free-living nitrogen-fixing microorganisms cover a morphological range that goes from unicellular organisms such as bacteria and some cyanobacteria, to multicellular, filamentous, therefore it is important to know how they behave and can be known by makinga microbial growth curve. For this study, 4 bubbled column photobioreactors with Fischerella TB22inoculum were prepared, they were placed in constant aeration with 12 hours of light and 12 hours of darkness for 40 days with different treatments of volumeadjustment of the culture medium and pH adjustment. The objective of this work was to evaluate the biomass growth by dry weight, optical density, pH, and ammonia of Fischerella sp. in the BG110 culture medium for 12 days. The variables that were measured from the growth curve of cyanobacteria followed the pattern of a typical microbial growth curve. (AU)

Diagnóstico do tratamento de esgotos no Rio Grande do Sul e sua associação aos riscos com a saúde / Diagnosis of sewage treatment in Rio Grande do Sul and its association with health risks

O presente trabalho tem como objetivo estimar o volume de esgoto gerado no RS em Demanda Bioquímica de Oxigênio (DBO kg/dia), e o volume de esgoto (DBO kg/dia) que chega até as estações de tratamento de esgoto ETEs; diagnosticar a eficiência das estações de tratamento de esgotoem relação ao percentual de remoção de (DBO kg/dia); correlacionando com a presença de cianobactérias próximas aos pontos de captação de água superficial para consumo humano; analisar se há inativação ou remoção de protozoários no lançamento do efluente ao retornar ao manancial. Como resultado obteve-se o volume total de matéria orgânica lançada nos corpos hídricos por dia estimado no Rio Grande do Sul é 511.769,00 DBO kg/dia no RS. As Estações de Tratamento de Esgoto captam 390.285,45 DBO kg/dia apresentaram resultados de remoção de 145.500,50 DBO kg/dia e de 22.963,21 DBO kg/dia residual; O volume ocioso das ETEs fica em torno de 5.049,89 L/s. O processo de eliminar ou inativar grande quantidade de protozoários presentes no afluente se dá através de etapas terciárias avançadas nas ETEs. Não encontramos no RS no ano 2019 ETEs com esses tipos avançados de tratamento. Sugere-se então que haja um melhor aproveitamento destas estações de tratamento no que se refere à quantidade e qualidade de volume de esgoto tratado e que sejam ampliados estudos para viabilidade de projetos de execução de novas ETEs para os demais municípios que despejam seus esgotos in natura nos mananciais impactando-os.

This work aims to estimate the volume of sewage generated in RS in Biochemical Oxygen Demand (BOD kg/day), and the volume of sewage (BOD kg/day) that reaches the sewage treatment stations (STS)s; diagnose the efficiency of sewage treatment station in relation to the percentage of removal (BOD kg/day); correlating with the presence of cyanobacteria close to the surface water intake points for human consumption; analyze if there is inactivation or removal of protozoa in the discharge of the effluent when returning to the source. As a result, the total volume of organic matter released into water bodies per day estimated in Rio Grande do Sul is 511,769.00 BOD kg/day in RS. The Sewage Treatment Plants capture 390,285.45 BOD kg/day showed removal results of 145,500.50 BOD kg/day and 22,963.21 BOD kg/day residual; The idle volume of ETEs is around 5,049.89 L/s. The process of eliminating or inactivating a large amount of protozoa present in the tributary takes place through advanced tertiary stages in the STSs. We did not find in RS in the year 2019 STSs with these advanced types of treatment. It is therefore suggested that there is a better use of these treatment plants with regard to the quantity and quality of the volume of treated sewage and that studies for the feasibility of projects for the execution of new STSs be expanded to the other municipalities that dump their raw sewage in water sources impacting them.

Zika virus syndrome, lack of environmental policies and risks of worsening by cyanobacteria proliferation in a climate change scenario / Síndrome do zika vírus, carência de políticas ambientais e riscos de agravamento por proliferação de cianobactérias em um cenário de mudanças climáticas

ABSTRACT Almost half of the Brazilian population has no access to sewage collection and treatment. Untreated effluents discharged in waters of reservoirs for human supply favor the flowering of cyanobacteria - and these microorganisms produce toxins, such as saxitoxin, which is a very potent neurotoxin present in reservoirs in the Northeast region. A recent study confirmed that chronic ingestion of neurotoxin-infected water associated with Zika virus infection could lead to a microcephaly-like outcome in pregnant mice. Cyanobacteria benefit from hot weather and organic matter in water, a condition that has been intensified by climate change, according to our previous studies. Considering the new findings, we emphasize that zika arbovirus is widespread and worsened when associated with climate change, especially in middle- or low-income countries with low levels of sanitation coverage.

RESUMO No Brasil, quase metade da população não tem acesso a coleta e tratamento de esgotos. Os efluentes não tratados, devido a contaminação das águas de reservatórios para abastecimento humano, geram florações de cianobactérias - e esses microrganismos produzem toxinas, como a saxitoxina, uma neurotoxina bastante potente e presente nos reservatórios da região Nordeste. Estudo recente confirmou que a ingestão crônica de água contaminada com neurotoxinas associada a infeção pelo zika vírus em camundongos prenhes poderia levar a desfecho semelhante a microcefalia. Cianobactérias são beneficiadas por tempo quente e matéria orgânica na água, e essas condições estão sendo intensificadas pelas mudanças climáticas, segundo nossos estudos anteriores. Aqui, ressaltamos que, à luz dos novos achados, a arbovirose do zika, associada às mudanças do clima, se amplifica e se agrava, sobretudo em países de média ou baixa renda, com baixos níveis de cobertura de saneamento.

Cianobacterias en las playas: riesgos toxicológicos y vulnerabilidad infantil / Cyanobacteria in beaches: toxicological risk and child vulnerability / Cianobactérias nas praias: riscos toxicológicos e vulnerabilidade infantil

Resumen: Las cianobacterias son bacterias fotosintéticas del plancton de aguas dulces y saladas. Su acumulación excesiva en las aguas recreacionales y potables se conoce como floración algal, con potenciales efectos en la salud de seres humanos. Son predominantemente estivales, en cursos de aguas dulces y pueden alcanzar las costas oceánicas. Esta es una revisión bibliográfica basada en las publicaciones registradas en PubMed-Medline, BVS (Biblioteca Virtual en Salud que incluye SciELO, LILACS, IBECS), Springer, Science Direct, Portal Timbó, y obtenidas con los términos "cianobacterias", "cianotoxinas", "microcistina", "floraciones algales", "pediatría", "niños", "recreacional" "intoxicación" "exposición", sin límites de año de publicación. Su objetivo es actualizar el conocimiento sobre cianotoxinas y efectos en la salud humana, analizar escenarios de riesgo de exposición a cianobacterias, particularmente en niños, y reforzar medidas preventivas de la exposición y promover acciones desde el sector sanitario. Las manifestaciones clínicas de la intoxicación por exposición reciente son malestar general, irritación de piel y mucosas, síntomas respiratorios e incluso afectación hepática en casos graves. Es preciso incluir estas floraciones en la etiopatogenia de estos cuadros y relacionarlos con la exposición utilizando la monitorización de floraciones del país. La exposición a cianobacterias es un riesgo emergente para la salud. La evidencia científica de los últimos años consolida el conocimiento de efectos en la salud humana por exposición a cianobacterias. Recientemente se identifica el baño, juegos infantiles e ingesta de arena en costas con altos niveles de contaminación como un escenario de riesgo en niños. La evidencia de hepatoxicidad por cianotoxinas es aún más escasa, aunque existen casos documentados. El sistema de monitoreo de playas permite a los equipos de salud incorporar la sospecha de exposición a cianobacterias y el posible contacto con sus toxinas, para poder realizar un diagnóstico temprano y participar en la prevención de la exposición.

Summary: Cyanobacteria are photosynthetic bacteria from fresh and saltwater plankton. Its excessive accumulation in recreational and drinking waters is known as algal bloom that could potentially affect human health. They are mainly seen in summer, in freshwater waterways and can reach the ocean coasts. This is a bibliographic review based on the publications registered in PubMed-Medline, BVS (Virtual Health Library that includes SciELO, LILACS, IBECS), Springer, Science Direct, Portal Timbó, and obtained with the terms "cyanobacteria", "cyanotoxins", "Microcystine", "algal blooms", "pediatrics", "children", "recreational" "intoxication" "exposure" without year of publication limits. The review aims to update knowledge about cyanotoxins and their effect on human health; analyze risk scenarios of exposure to cyanobacteria, particularly in children, and reinforce preventive measures for exposure, as well as to promote actions from the health sector. The clinical manifestations of recent exposure poisoning are general malaise, skin and mucous irritation, respiratory symptoms and even liver insufficiency in severe cases. It is necessary to include these blooms in the pathogenesis of these clinic cases and relate them to the exposure using the monitoring of blooms in the country. Exposure to cyanobacteria is an emerging health risk. The scientific evidence of the last years consolidates knowledge on the effect of exposure to cyanobacteria on human health. Recently, bathing, children's games, and sand intake on coasts with high levels of contamination have been identified as a risk scenario for children. Evidence of cyanotoxin hepatoxicity is even scarcer, although there are documented cases. The beach monitoring system allows health teams to incorporate the suspicion of exposure to cyanobacteria and possible contact with their toxins, to make an early diagnosis and participate in the prevention of exposure.

Resumo: As cianobactérias são bactérias fotossintéticas do plâncton de águas doces e salgadas. Sua acumulação excessiva em águas recreacionais e potáveis é conhecida como floração algal com potenciais efeitos sobre a saúde de seres humanos. São predominantemente estivais, ocorrem em cursos de águas doces e podem alcançar as costas oceânicas. Esta é uma revisão bibliográfica baseada nas publicações indexadas em PubMed-Medline, BVS (Biblioteca Virtual em Saúde que inclui SciELO, LILACS, IBECS), Springer, Science Direct, Portal Timbó, e obtidas com os termos "cianobactérias", "cianotoxinas", "microcistina", "florações algais", "pediatria", "crianças", "recreacional" "intoxicação" "exposição", sem limites de ano de publicação. Seu objetivo é atualizar o conhecimento sobre cianotoxinas e efeitos sobre a saúde humana; analisar cenários de risco de exposição a cianobactérias, particularmente em crianças e reforçar medidas preventivas de exposição e promover ações do setor sanitário. As manifestações clínicas da intoxicação por exposição recente são mal-estar geral, irritação de pele e mucosas, sintomas respiratórios e incluso afetação hepática nos casos graves. É preciso incluir estas florações, na etiopatogenia destes quadros e relacioná-las com a exposição utilizando o monitoramento das florações do país. A exposição a cianobactérias é um risco emergente para a saúde. A evidência científica dos últimos anos consolida o conhecimento sobre os efeitos sobre a saúde humana por exposição a cianobactérias; recentemente foram identificados como um cenário de risco para crianças: o banho, jogos infantis e ingestão de areia nas costas com altos níveis de contaminação. A evidência de hepatoxicidade por cianotoxinas é ainda escassa embora existam casos documentados. O sistema de monitoramento de praias permite as equipes de saúde incorporar a suspeita de exposição a cianobactérias e o possível contacto com suas toxinas, para poder realizar um diagnóstico precoce e participar na prevenção da exposição.

Dinámica del consumo y aporte de nutrientes de fitoplancton, dominado por Microcystis sp. (Cyanophyceae) del lago de Amatitlán / Dynamics of the consumption and nutrient supply of phytoplankton, dominated by Microcystis sp. (Cyanophyceae) of Lake Amatitlán

El impacto antrópico cercano al lago de Amatitlán ha generado niveles altos de eutrofización que conllevan cambios en la dinámica del ecosistema. Uno de ellos es la proliferación de cianobacterias del género Microcystis que pueden llegar a ser perjudiciales para la fauna y flora de lugar e incluso para los seres humanos. Se presenta el caso de cultivo de un consorcio de fitoplancton, tomado directamente del lago y llevado al laboratorio en condiciones controladas, para medir su consumo y aporte de nitrógeno y fósforo, además de los cambios en los factores fisicoquímicos y la biomasa. Se observó la presencia de diatomeas del genero Nitszchia y cianobacterias como Dolichospermum, con una marcada dominancia de Microcystis sp. Se analizó el porcentaje de cambio en la concentración de nutrientes. Los resultados indican que hubo un aumento en las concentraciones de amonio, nitrato y nitrógeno inorgánico disuelto, mientras que disminuyó el nitrógeno total, el fósforo total y los ortofosfatos. Esto indica que hay aporte de nitrógeno inorgánico, consumo de fósforo y nitrógeno orgánico. El fósforo parece ser el nutriente limitante, ya que, al consumirse en un 90 % la biomasa empieza a decrecer.

The anthropic impact near lake Amatitlán has generated high levels of eutrophication that lead to changes in ecosystem dynamics. One of them is the proliferation of cyanobacteria of the genus Microcystis that can be harmful to the fauna and flora of the place and even to humans. The case of cultivation of a phytoplankton consortium, taken directly from the lake and taken to the laboratory under controlled conditions, to measure its consumption and contribution of nitrogen and phosphorus, in addition to changes in physicochemical factors and biomass is presented. The presence of diatoms of the genus Nitszchia and cyanobacteria such as Dolichospermum was detected, with a marked dominance of Microcystis sp. The percentage change in nutrient concentration was analyzed. The results indicated that there was an increase in the amounts of dissolved inorganic ammonium, nitrate and nitrogen, while total nitrogen, total phosphorus and orthophosphates decreased. This indicates that there is contribution of inorganic nitrogen, consumption of phosphorus and organic nitrogen. Phosphorus seems to be the limiting nutrient, since, when consumed by 90 %, biomass begins to decrease.

Estratégias para identificação e quantificação de aminoácidos tipo micosporinas (MAAs) em cianobactérias e outros organismos para aplicação em cosmética e farmacêutica / Identification and quantification methods of mycosporine-like amino acids (MAAs) in cyanobacteria and other microorganisms for cosmetic and pharmaceutical application

Os aminoácidos tipo micosporinas (MAAs) são compostos, produzidos por algumas espécies de cianobactérias e outros microorganismos, principalmente quando são expostos a radiação ultravioleta (UVR). Estes compostos, que vêm demonstrando funções fotoprotetoras e antioxidantes, têm sido pesquisados para aplicação em protetores solares e em produtos antienvelhecimento. O presente estudo focou na caracterização de cianobactérias e outros organismos quanto à produção de MAAs com potencial aplicação em cosméticos. Neste estudo foram desenvolvidos diversos métodos para identificação (via HPLC-DAD-MS/HRMS), purificação (via HPLC-DAD) e quantificação de MAAs (via LC-MS/MS). Pelo método de identificação de MAAs verificou-se que, das 75 cianobactérias estudadas, 27 cepas (38%) sintetizam MAAs. A cepa Oscilatoria sp. CMMA 1600 produziu a maior diversidade de MAAs. 10 MAAs diferentes foram identificados incluindo um MAA de massa molecular 316 Da. Através de dados espectroscópicos obtidos via HPLC-DAD-HRMS e RMN 1D e 2D confirmou-se que se tratava da micosporina-glicina-alanina. A biossíntese natural deste composto por cianobactérias foi relatada pela primeira vez neste estudo. Quanto à quantificação de MAAs, o protocolo de extração otimizado possibilitou uma excelente recuperação dos compostos de interesse, além de ser bastante simples e não utilizar solventes poluentes. As análises via LC-MS/MS foram realizadas através de experimentos de MRM em modo positivo usando uma coluna de fase reversa. O método validado permitiu determinar e quantificar com precisão os MAAs porphyra-334, shinorina e micosporina-glicina-alanina em corridas de apenas 6 minutos, com limites de deteção inferiores a 0,005 µg.mg -1. Aplicando o método de LC-MS/MS realizaram experimentos de indução de MAAs através de exposição à UVR tendo-se observado um aumento da concentração de MAAs nas cepas que já sintetizam estes compostos e, outras cepas começaram a produzir pelo menos um MAA. As cepas de S. torques-reginae (ITEP-024 e ITEP-026) produziram a maior concentração de MAAs. A cepa ITEP-024 foi ainda exposta a diferentes radiações tendo-se observado que a UVB é que mais influencia a produção de MAAs. Neste estudo foi demonstrado o potencial das cianobactérias como produtores de MAAs que podem ser utilizados como fotoprotores em protetores solares

Mycosporines and mycosporine-like amino acids (MAAs) are UV-absorbing compounds produced by cyanobacteria and other organisms, especially upon exposer to solar ultraviolet radiation (UVR). These compounds are photoprotective and some have additional antioxidant functíons useful to the natural cosmetics market. This study aims to identify MAAs-producing cyanobacteria with potential applicatíons in cosmetics. A HPLC-DAD-MS/HRMS method for the identification of MAAs was developed. Out of the 75 cyanobacteria studied, 27 strains (38%) synthesized MAAs. Oscilatoria sp. CMMA 1600, from homocyte type, produced the greatest diversity of MAAs. 10 different MAAs were identified including a MAA with molecular weight of 316 Da. The chemical structure of mycosporine-glycine-alanine was confirmed by 1D/2D NMR and HRMS analyses. This compound has never been reported from a natural source. In this study, a validated LC-MS/MS quantification method for MAAs is also presented. An easy-to-handle and rapid extraction procedure was developed which uses only water and volatile additives as the extractor solvents. The LC-MS/MS method was performed using multiple reaction monitoring in positive mode with a reverse-phase column. The method enabled the accurate determination and quantification of the MAAs porphyra-334, shinorine and mycosporine-glycine-alanine in a 6 minutes running time, with limits of detection < 0.005 µg.mg-1. MAAs induction experiments were performed through UVR exposure. MAAs are constitutively produced by some cyanobacteria and production was further enhanced following UVirradiance. Other strains start to produce at least one MAA after UV-irradiance. Sphaerospermopsis torques-reginae strain (ITEP-024 and ITEP-026) produced the highest concentration of these photoprotective compounds. S. torques-reginae ITEP 024 strain was further exposed to different radiation compositíons. MAAs were significantly influenced by UVB. In this study, the potential of cyanobacteria as MAA producers, that can be used as photoprotectors in sunscreens, has been demonstrated

Población de cianobacterias en suelos arroceros: efecto de las rotaciones / Cyanobacteria populations in rice soils: rotation effects

Este trabajo tuvo como objetivo evaluar el efecto de rotaciones arroceras sobre la población de cianobacterias durante el ciclo de cultivo. El estudio se condujo en Entre Ríos (Argentina). Se evaluaron cuatro rotaciones: arroz-soja (AS); arroz-soja-maíz-soja (ASMS): arroz-arroz (AA) y arroz-pradera (AP). Muestras de suelo y agua se tomaron en cuatro estados del cultivo: implantación, macollaje, panoja embuchada y madurez fisiológica. El número potencial de cianobacterias fue menor en relación a otras áreas arroceras. Los menores recuentos se registraron en el período de implantación y los mayores en el de panoja embuchada. El análisis estadístico no mostró efecto de las rotaciones sobre el número de cianobacterias, aunque AA presentó la mayor abundancia. Se reconocieron un total de 13 géneros de cianobacterias Lyngbya, Oscillatoria,Plectonema, Spirulina, Anabaena, Nostoc, Aulosira, Calothrix, Gloeotri-chia, Aphanocapsa,Croococcus, Mycrocystis y Gloeocapsa. La rotación AP registró la riqueza de géneros.Las rotaciones AS y AA presentaron mayor proporción de cianobacterias filamentosas. Los valores de biodiversidad de Simpson fueron bajos. Se concluyó que las rotaciones arroceras evaluadas tuvieron efecto sobre la proporción de cianobacterias unicelulares a filamentosas, los géneros presentes y la riqueza.

The aim of this study was to evaluate the effect of different rice rotations on cyanobacteria populations during the growing season. The study was conducted in Entre Ríos (Argentina). Four crop rotations were evaluated: rice-soybean (AS); rice-soybean-corn-soybean (ASMS); rice-rice (AA) and rice-pasture (AP). Soil and water samples were collected in 4 stages: implantation, tillering, booting and physiological maturity. The potential number of cyanobacteria population was lowerin relation to other rice sites previously reported. The lowest counts were recorded during implantation whereas the highest counts were obtained during booting. The statistical analysis showed no effect of rotation on the variable number of cyanobacteria, although AA was the most abundant. A total of 13 taxa were identified: Lyngbya, Oscillatoria, Plectonema, Spirulina, Anabaena, Nostoc, Aulosira, Calothrix, Gloeotrichia, Aphanocapsa, Croococcus, Mycrocystis and Gloeocapsa. AP rotation recorded the lowest number of genera. AS and AA had a higher proportion of filamentous cyanobacteria. Simpson's index was low. It was concluded that crop rotations had a differential effect on some aspects of cyanobacteria population such as the proportion of unicellular to filamentous cyanobacteria, genera presence and richness.

Sodium and nitrate favor the steady state of cyanobacteriain a semiarid ecosystem

This study characterized steady state events (SS) in a semiarid reservoir (Brazil) dominated by cyanobacteria for 130 weeks, and evaluated the influence of different abiotic variables on their occurrence. It was hypothesized that, in semiarid reservoirs, steady state events are independent from seasonality and influenced by high nutrient concentrations, and steady state periods show low variation of species. Our data revealed the occurrence of SS events in both the dry and wet seasons. Higher concentrations of nitrate and sodium together with higher values of color favored the development of SS. Species composing the SS dominated by Microcystis aeruginosa (Kützing) Kützingwas correlated with higher sulfate concentrations, while higher values of turbidity and sodium concentrations favored the SS of Cylindrospermopsis raciborskii (Woloszynska) Seenaya and Suba Rajuand Planktothrix agardhii (Gomont) Anagnostidis and Komárek. The results supported the hypothesis and showed the importance of variables rarely evaluated in works of this nature, such as sodium and sulfate in SS establishment of cyanobacteria.

Progress in the molecular mechanism of KaiA regulating cyanobacterial circadian clock / 生物工程学报

The cyanobacterial circadian clock has three relatively independent parts: the input path, the core oscillator, and the output path. The core oscillator is composed of three clock proteins: KaiA, KaiB, and KaiC. The interactions among these three proteins generate a rhythmic signal and convey the input signals to the output signals to maintain the accuracy and stability of the oscillation of downstream signals. Based on the cyanobacterial circadian clock and the structure, function, and interaction of the clock proteins of the core oscillator, combining the recent results from our laboratory, this review summarized the recent progresses of the molecular mechanism of KaiA in regulating KaiC's enzymatic activity, mediating phase reset of the oscillator, and competing with CikA for the binding site of KaiB.

Cyanobacteria based photosynthetic production of sucrose: development and prospect / 生物工程学报

Biorefinery technologies provide promising solutions to achieve sustainable development facing energy and environment crisis, while abundant sugar feedstock is an essential basis for biorefinery industries. Photosynthetic production of sucrose with cyanobacteria is an alternative sugar feedstock supply route with great potentials. Driven by solar energy, cyanobacteria photosynthetic cell factory could directly convert carbon dioxide and water into sucrose, and such a process could simultaneously reduce carbon emissions and supply sugar feedstocks. Here we introduced the history and updated the state-of-the-art on development of cyanobacteria cell factories for photosynthetic production of sucrose, summarized the progress and problems on mechanisms of sucrose synthesis, metabolic engineering strategies and technology expansions, and finally forecasted the future development direction in this area.