Genetic polymorphism in brazilian microcystis spp. (Cyanobacteria) toxic and non-toxic through RFLP-PCR of the cpcBA-IGS

The escalating occurrence of cyanobacterial toxic blooms demands a better understanding of genetic variability as an auxiliary expedient in species identification, collaborating with the monitoring of water destined to public supply. This study aimed at the unraveling of genetic polymorphism in the toxic and nontoxic strains of Microcystis (Cyanobacteria) species, isolated from diverse Brazilian localities through the RFLP-PCR technique applied to the c-phycocyanin encoding operon and its intergenic spacer (cpcBA-IGS). Eighteen strains belonging to M. aeruginosa, M. panniformis, M. protocystis and M. wesenbergii, plus two other unidentified strains, were analyzed by means of the morphological and molecular data. The molecular data constituted three groups with low similarity values unrelated to the geographical origin, toxicity or morphospecies. A high genetic variability among the studied populations was unveiled by the results. Brazilian populations of Microcystis spp. displayed high genetic diversity when compared to those from Australia, Japan, United States and Europe. This ample genetic diversity could be observed through the diverse eletrophoretic profiles obtained among the strains from a single species. The presence of toxic and non-toxic strains was observed in the same species, as M. aeruginosa.A ocorrência de florações de cianobactérias tóxicas demanda um melhor entendimento da variabilidade genética como um instrumento auxiliar na identificação de espécies colaborando, assim, com o monitoramento de águas destinadas ao abastecimento público. Este estudo objetivou o conhecimento do polimorfismo genético de linhagens tóxicas e não tóxicas de espécies de Microcystis (Cyanobacteria), isoladas de diversas localidades brasileiras, utilizando a técnica molecular RFLP-PCR para o operon que codifica para a c-ficocianina e seu espaçador intergênico (cpcBA-IGS). Foram analisadas dezoito linhagens pertencentes as espécies Microcystis aeruginosa, M. panniformis, M. protocystis, M. wesenbergii e duas outras não identificadas através de dados morfológicos e moleculares. Os resultados moleculares formaram três agrupamentos com baixos valores de similaridade entre si os quais não foram relacionados à origem geográfica, toxicidade ou morfoespécies. As populações brasileiras de Microcystis spp. apresentaram alta diversidade genética quando comparadas com as da Austrália, Japão, Estados Unidos e Europa. Esta ampla diversidade genética pode ser vislumbrada através de diversos perfis eletroforéticos obtidos entre linhagens de uma mesma espécie. Nós encontramos a presença de linhagens tóxicas e não tóxicas em uma mesma espécie, como em M. aeruginosa.CAPES/PPGB/UFRPE(FAPESP) São Paulo Research Foundation(CNPq) Brazilian Council for Research and Developmen

Structure and dynamics of phytoplankton community in the Botafogo reservoir-Pernambuco-Brazil

The aim of the present study was to investigate the structure and dynamics of the phytoplankton in the Botafogo reservoir-PE-Brazil. Phytoplankton assemblages were identified from current literature and density was estimated using an inverted microscope. Concurrently to the sampling of biotic variables, measurements of abiotic parameters, such as water temperature, dissolved oxygen and pH, were determined using field probes and transparency was determined with a Secchi disk. Total phosphorus and total nitrogen concentrations were determined in laboratory. A total of 24 taxa were identified. Chlorophyta presented the greatest number of species. Species diversity in the reservoir was low throughout the study period. Principal component analysis revealed that Trachelomonas volvocina, Chlorella vulgaris, Euglena sp. and Peridinium gatunense were directly correlated with oxygen, turbidity and total nitrogen; Planktosphaeria gelatinosa, P. gatunense and Euglena sp. were directed correlated with total nitrogen; rainfall explained the occurrence of Monoraphidium arcuatum and Chlorella vulgaris.O objetivo do presente estudo foi investigar a estrutura e dinâmica do fitoplâncton no reservatório de Botafogo-PE-Brasil. A comunidade fitoplanctônica foi identificada com literatura atualizada e a densidade estimada usando microscópio invertido. Concomitantemente as coletas das variáveis bióticas, foram medidos alguns parâmetros abióticos como temperatura da água, oxigênio dissolvido, condutividade e pH usando sondas de campo e transparência com disco de Secchi. Concentrações de fósforo total e nitrogênio total foram determinados em laboratório. Vinte e quatro táxons foram identificados tendo Chlorophyta apresentado maior número de espécie. A diversidade de espécie no reservatório foi baixa durante todo o período de estudo. A análise de componentes principais mostrou que Trachelomonas volvocina, Chlorella vulgaris, Euglena sp. e Peridinium gatunense esteve diretamente correlacionada com oxigênio dissolvido, turbidez e nitrogênio total. P. gelatinosa, Peridinim gatunense e Euglena sp. estiveram diretamente relacionada com nitrogênio total; a chuva explicou a ocorrência de Monoraphidium arcuatum e Chlorella vulgaris.(CNPq) Brazilian Council for Research and Developmen

Acute toxicity of Microcystis spp. (Cyanobacteria) bloom on Moina minuta (Cladocera) in a tropical reservoir, Northeastern Brazil

Worldwide cyanobacterial blooms have been registered, where harmful species dominance is associated to producing of toxic compounds (cyanotoxins) with adverse effects on several organisms. Acute toxicity of crude extracts from Microcystis bloom occurring in Mundaú reservoir was evaluated by bioassays with the neotropical Cladocera Moina minuta. Samples were taken in the reservoir during the rainy (April/2012) and dry season (September/2012). Cyanobacterial analyses were performed by identification on optical microscopy and direct counting using an inverted microscope. Bloom samples were frozen, lyophilized and re-suspended in deionized water for preparation of extracts. Tests with the cladoceran were carried out in test tubes with different concentrations of the crude extract, diluted in 10 mL reconstituted water. In both rainy and dry periods, densities of Microcystis spp. were above 15x103 ind mL-1. Microcystin concentrations in the extracts were 0.70 (+ 0,009) (rainy season) and 0.69 (+ 0,005) (dry season) μg g-1. The LC50 (48h) of crude extract for both rainy and dry periods was 160 (100 – 255) and 72 (4 – 1113) mg L-1, respectively. These results indicated that extracts of Microcystis spp. were acutely toxic to M. minuta population with suggesting that such events represent potential toxicity to zooplankton

Microalgas de dois ambientes lóticos amazônicos, Rondônia, BrasilMicroalgae of two lotic amazonian environments, Rondônia, Brasil

O objetivo deste trabalho foi identificar microalgas de dois ambientes lóticos amazônicos, localizados nas bacias dos rios Purus e Madeira, em Porto Velho, Rondônia, Brasil. As coletas foram realizadas nos meses de setembro e novembro de 2009 (águas baixas) e de fevereiro e abril de 2010 (águas altas), nas subacias dos rios Açuã e Aponiã. As amostras foram coletadas através de arrastos verticais e horizontais na área marginal dos rios com rede de plâncton de 25mm de abertura de malha e preservadas em solução de Transeau. Simultaneamente, foram coletadas amostras para medições de pH, turbidez e condutividade elétrica. Em laboratório, foram confeccionadas lâminas e observadas em microscópio e a identificação das espécies através de literatura especializada. O período de águas baixas caracterizou-se, em geral, por pH de levemente ácido a neutro, se comparado ao período de águas altas em ambos os rios. Já a turbidez e condutividade elétrica foram menores nas águas altas em ambos os rios e, no rio Aponiã, nas águas baixas. Foram identificados 79 táxons distribuídos em cinco divisões taxonômicas, sete classes, 15 ordens e 25 famílias. Nove espécies ocorreram somente no período de estiagem e 25 espécies apenas no período chuvoso. As espécies com maior frequência de ocorrência foram Oscillatoria principes, Desmidium grevillei, Actinella sp., Bambusina brebisonii, Micrasterias rotata, Eunotia sp., Cladophora sp. e Spirogyra sp. Dos 79 táxons, 61 espécies são citadas pela primeira vez para o estado de Rondônia.AbstractThe aim of this study was to identify the microalgae of two lotic Amazonian environments, situated in the watershed of Purus and Madeira rivers, in Porto Velho, Rondonia, Brazil. The sampling was performed in September and November of 2009 (low water) and February and April of 2010 (high water) in the sub-water basins of Acua and Aponiã rivers. The samples were collected by vertical and horizontal hauls in the marginal area of the rivers with a 25 μm nylon plankton net and preserved in Transeau solution. Simultaneously it was collected samples for pH, turbidity and conductivity. In laboratory, slides were prepared and observed under a microscope and species were identified based on specialized literature. The study revealed that the low water period was characterized generally by pH slightly acid to neutral, as well as turbidity and lower electrical conductivity as compared to the period of high water levels in both rivers. It was identified 79 taxa in five taxonomic divisions, seven classes, 15 orders and 25 families. Nine species occurred only in the dry period and only 25 species in the rainy period. The species with the highest frequency of occurrence were Oscillatoria principes, Desmidium grevillei, Actinella sp., Bambusina brebisonii, Micrasterias rotata, Eunotia sp., Cladophora sp. e Spirogyra sp. Of the 79 taxa, 61 species are first cited for the state of Rondônia

Variação temporal da comunidade fitoplanctônica em curtos intervalos amostrais no reservatório de Mundaú, Nordeste brasileiro

The aim of this study was to determine how abiotic factors drive the phytoplankton community in a water supply reservoir within short sampling intervals. Samples were collected at the subsurface (0.1 m) and bottom of limnetic (8 m) and littoral (2 m) zones in both the dry and rainy seasons. The following abiotic variables were analyzed: water temperature, dissolved oxygen, electrical conductivity, total dissolved solids, turbidity, pH, total nitrogen, nitrite, nitrate, total phosphorus, total dissolved phosphorus and orthophosphate. Phytoplankton biomass was determined from biovolume values. The role abiotic variables play in the dynamics of phytoplankton species was determined by means of Canonical Correspondence Analysis. Algae biomass ranged from 1.17×10(4) to 9.21×10(4) µg.L-1; cyanobacteria had biomass values ranging from 1.07×10(4) to 8.21×10(4) µg.L-1. High availability of phosphorous, nitrogen limitation, alkaline pH and thermal stability all favored cyanobacteria blooms, particularly during the dry season. Temperature, pH, total phosphorous and turbidity were key factors in characterizing the phytoplankton community between sampling times and stations. Of the species studied, Cylindrospermopsis raciborskii populations were dominant in the phytoplankton in both the dry and rainy seasons. We conclude that the phytoplankton was strongly influenced by abiotic variables, particularly in relation to seasonal distribution patterns.O objetivo deste estudo foi detectar a influência dos fatores abióticos sobre a comunidade fitoplanctônica em um reservatório de abastecimento, em curtos intervalos amostrais. As amostras foram coletadas na subsuperfície (0,1 m) e fundo das regiões pelágica (8 m) e litorânea (2 m), durante dois períodos sazonais, estações seca e chuvosa. As variáveis abióticas analisadas foram: temperatura da água, oxigênio dissolvido, condutividade elétrica, sólidos totais dissolvidos, turbidez, pH, nitrogênio total, nitrito, nitrato, fósforo total, fósforo total dissolvido e ortofosfato. A biomassa fitoplanctônica foi determinada a partir de valores de biovolume. A influência das variáveis abióticas na dinâmica das espécies fitoplanctônicas foi determinada através da Análise de Correspondência Canônica. A biomassa algal variou de 1,17×10(4) a 9,21×10(4) µg.L-1, sendo que as cianobactérias apresentaram valores de biomassa entre 1,07×10(4) e 8,21×10(4) µg.L-1. A mais alta disponibilidade de fósforo, limitação de nitrogênio, pH alcalino e estabilidade térmica favoreceram florações de cianobactérias, especialmente durante o período seco. As diferenças entre os horários e as estações amostrais na distribuição da comunidade fitoplanctônica foram determinadas pelos valores de temperatura, pH, fósforo total e turbidez. As associações fitoplanctônicas que mais se destacaram foram do grupo funcional Sn, formado por Cylindrospermopsis raciborskii, que predominou no sistema em ambos períodos sazonais. Concluiu-se que o fitoplâncton apresentou uma forte influência das variáveis abióticas especialmente no estabelecimento de padrões sazonais de distribuição.Conselho Nacional de Desenvolvimento Científico (CNPq

Temporal and spatial dynamics of phytoplankton near farm fish in eutrophic reservoir in Pernambuco, Brazil

Spatial and temporal variations in phytoplankton communities in continental waters have received attention from limnologists, since they are differently influenced by many physico-chemical and biological fac- tors. This study was undertaken with the aim to identify the environmental variables that influence the temporal and spatial dynamics of the phytoplankton near a fish farm in the Jucazinho reservoir, in a semi-arid region of Northeastern Brazil. Samples were taken from three sampling sites, at two depths during the rainy (Aug 2008, Feb and Mar 2009) and dry (Oct, Nov and Dec 2008) seasons. Phytoplankton was identified, density determined, and biomass values obtained. Concomitantly, abiotic analyses were performed for the characterization of the system. The reservoir was homogeneous with regard to the spatial-temporal variation in hydrological variables: water well oxygenated at the surface and anoxic at the bottom; pH ranging from neutral to alkaline; temperatures always above 25oC; high turbidity; and high electrical conductivity at all sampling sites and both depths. For both seasons, there was limited nitrogen and high concentrations of phosphorus. Cyanophyta species were pre- dominant, generally representing 80% of the phytoplankton biomass throughout practically the entire study, at all sampling sites and both depths. Co-dominance of cyanobacteria belonging to H1, MP, S1 and Sn associations was recorded in most of the months studied, except August 2008, when there was a substitution of the S1 asso- ciation (Planktothrix agardhii) by the P association (Aulacoseira granulata). Water temperature, precipitation and pH were the parameters with the greatest influence over the temporal variation in phytoplankton, whereas the vertical distribution of the phytoplankton biomass was directly related to the availability of light in the water column. There were no spatial or temporal differences in water quality, likely due to the fact that the sampling sites were near to one another and received the same nutrients stemming from the rations given to the fish as well as their excrement. The predominance of cyanobacteria through the study was certainly due to the influence of the feeding products offered to and excreted from the fish, which eutrophicated the system. In temporal terms, the change in the algal structure is explained by the change in the physical conditions of the water between the surface and bottom, as well as the climatologic conditions, especially the change in wind direction, with the consequent fetch of the water.Las variaciones espaciales y temporales en las comunidades de fitoplancton en las aguas continentales han recibido la atención de limnólogos, ya que están influenciados de manera diferente por muchos factores físico-químicos y biológicos. El objetivo del presente trabajo fue identificar las variables ambientales que ejercieron influencia sobre la dinámica temporal y espacial de la comunidad fitoplanctónica cerca a una granja pisícola en la represa de Jucazinho, la cual se encuentra en la región semiárida brasilera. Las recolecciones fueron realizadas en tres estaciones de moni- toreo a dos profundidades, durante los periodos lluviosos (agosto 2008 y febrero, marzo 2009) y de sequía (octubre, noviembre y diciembre 2008). El fitoplancton fue identificado y su densidad determinada, y posteriormente convertida a biomasa. También se realizaron análisis abióticos para caracterizar el sistema. La represa resultó homogénea en cuanto a la variación espacio-temporal de las variables hidrológicas, agua oxigenada en la superficie y anóxica en el fondo, pH varía neutro-alcalino, temperaturas superiores a 25°C y conductividad y turbidez elevadas. En los dos periodos estacionales fueron observadas limitaciones de nitrógeno y elevadas concentraciones de fósforo. Las Cyanophytas fueron predominantes en el ambiente y representaron en general más del 80% de la biomasa fitoplanctónica durante prácticamente todo el periodo de estudio en todas las estaciones y profundidades de recolecta. La co-dominancia de cianobacterias pertenecientes a las asociaciones H1, MP, S1 y Sn, fue registrada durante la mayor parte del periodo de estudio, excepto en ago/2008, cuando ocurrió una sustitución de la asociación S1 (Planktothrix agardhii) por P (Aulacoseira granulata) en la represa. Los factores que tuvieron más influencia sobre la variación temporal del fitoplancton fueron temperatura del agua, precipitación pluvial y pH. La distribución vertical de la biomasa fitoplanctónica estuvo directamente relacionada con la disponibilidad de luz en la columna de agua. No se encontraron diferencias espaciales ni temporales en la calidad del agua, seguramente porque los putos de muestreo eran cercanos y recibieron la misma carga de nutrientes provenientes del concentrado de los peces y de la excreción de estos mismos. La predominancia de cianobacterias a lo largo de todo el estudio seguramente fue debida a la influencia de los productos del alimento suministrado a los peces y sus excrementos que eutrifican el sistema. Temporalmente, el cambio en la estructura de las algas es explicado por la modificación de las condiciones físicas del agua entre superficie y fondo, provocado por el cambio en la dirección del viento con la consecuente alteración del fetch del agua

Spatial-temporal variation in coiled and straight morphotypes of Cylindrospermopsis raciborskii (Wolsz) Seenayya et Subba Raju (Cyanobacteria) Variação espaço-temporal dos morfotipos espiralado e reto de Cylindrospermopsis raciborskii (Wolsz) Seenayya et Subba Raju (Cyanobacteria)

This study reports the spatial and temporal behavior of straight and coiled morphotypes of C. raciborskii in a reservoir in Brazil's semi-arid region as well as the main factors responsible for the variability. Two set of samples were collected from the subsurface and bottom in the central region of a reservoir in two seasonal periods (dry - January 2005; rainy - June 2005) over 20-hour sampling periods during daylight (8 am, 12 pm and 4 pm) and dark (8 pm, 12 am and 4 am) hours. Measurements of abiotic parameters were determined concurrently to the sampling of biotic variables. Two C. raciborskii morphotypes were found in the reservoir: straight and coiled. There was no difference in density of the straight and coiled C. raciborskii morphotypes between the different sampling times for either season. Vertical differences were found in the distribution of both morphotypes in both seasons, with greater densities recorded at the subsurface. Densities of the two C. raciborskii morphotypes were greater in the dry season, with the density of the coiled morphotype at the surface two-fold greater than that of the straight morphotype and that found in the rainy season. The ecological success of the coiled morphotype was due to thermal stratification, whereas a mixed condition was determinant in the success of the straight morphotype.O presente estudo reporta o comportamento espacial e temporal dos morfotipos reto e espiralado de C. raciborskii em um reservatório da região semi-árida do Brasil bem como os principais fatores responsáveis pela variabilidade. Dois conjuntos de amostras foram coletadas na subsuperfície e próximo ao sedimento, na região central do reservatório, em dois períodos sazonais (seco - janeiro 2005; chuvoso - junho 2005), sendo realizadas amostragens de 20 horas durante horários claros (08hs, 12hs e 16hs) e escuros (20hs, 00hs e 04hs). Medidas de parâmetros abióticos foram realizadas concomitantes as amostragens das variáveis bióticas. Dois morfotipos de C. raciborskii foram encontrados no reservatório: reto e espiralado. Não houve diferenças na densidade dos morfotipos reto e espiralado de C. raciborskii entre os diferentes horários amostrais em cada período sazonal. Diferenças verticais foram encontradas na distribuição de ambos morfotipos em ambos períodos sazonais, com maiores densidades registradas na subsuperfície. As densidades dos dois morfotipos de C. raciborskii foram maiores no período seco, com densidade do morfotipo espiralado na subsuperfície duas vezes maior que o morfotipo reto e o encontrado no período chuvoso. O sucesso ecológico do morfotipo espiralado foi devido à estratificação térmica, enquanto as condições de mistura foram determinantes no sucesso do morfotipo reto