Changes in freshwater sediment microbial populations during fermentation of crude glycerol

BACKGROUND: This work studied how the exposure to an unusual substrate forced a change in microbial populations during anaerobic fermentation of crude glycerol, a by-product of biodiesel production, with freshwater sediment used as an inoculum. RESULTS: The microbial associations almost completely (99.9%) utilized the glycerol contained in crude glycerol 6 g L 1 within four days, releasing gases, organic acids (acetic, butyric) and alcohols (ethanol, n-butanol) under anaerobic conditions. In comparison with control medium without glycerol, adding crude glycerol to the medium increased the amount of ethanol and n-butanol production and it was not significantly affected by incubation temperature (28 C or 37 C), nor incubation time (4 or 8 d), but it resulted in reduced amount of butyric acid. Higher volume of gas was produced at 37 C despite the fact that the overall bacterial count was smaller than the one measured at 20 C. Main microbial phyla of the inoculum were Actinobacteria, Proteobacteria and Firmicutes. During fermentation, significant changes were observed and Firmicutes, especially Clostridium spp., began to dominate, and the number of Actinobacteria and Gammaproteobacteria decreased accordingly. Concentration of Archaea decreased, especially in medium with crude glycerol. These changes were confirmed both by culturing and culture-independent (concentration of 16S rDNA) methods. CONCLUSIONS: Crude glycerol led to the adaptation of freshwater sediment microbial populations to this substrate. Changes of microbial community were a result of a community adaptation to a new source of carbon.

Illumina sequencing and assessment of new cost-efficient protocol for metagenomic-DNA extraction from environmental water samples

Abstract In this study, the development and assessment of a modified, efficient, and cost-efficient protocol for mDNA (metagenomic DNA) extraction from contaminated water samples was attempted. The efficiency of the developed protocol was investigated in comparison to a well-established commercial kit (Epicentre, Metagenomic DNA Isolation Kit for Water). The comparison was in terms of degree of shearing, yield, purity, duration, suitability for polymerase chain reaction and next-generation sequencing in addition to the quality of next-generation sequencing data. The DNA yield obtained from the developed protocol was 2.6 folds higher than that of the commercial kit. No significant difference in the alpha (Observed species, Chao1, Simpson and PD whole tree) and beta diversity was found between the DNA samples extracted by the commercial kit and the developed protocol. The number of high-quality sequences of the samples extracted by the developed method was 20% higher than those obtained by the samples processed by the kit. The developed economic protocol successfully yielded high-quality pure mDNA compatible with complex molecular applications. Thus we propose the developed protocol as a gold standard for future metagenomic studies investigating a large number of samples.

Isolamento e caracterização de espécies de Pythium de ambientes aquáticos no Estado do Rio Grande do Sul e avaliação da patogenicidade em modelo experimental / Isolation and characterization of Pythium species from swampy areas in the Rio Grande do Sul, Brazil, and evaluation of pathogenicity in an experimental model

Foram coletadas 186 amostras de água de ambientes pantanosos em 13 municípios das regiões Sul, Central e Oeste do Rio Grande do Sul, Brasil, com o objetivo de isolar e caracterizar espécies de Pythium e avaliar a sua patogenicidade empregando coelhos como modelo experimental. Em 11,8% (n=22) das águas coletadas foram isoladas diferentes espécies de Pythium incluindo: P. insidiosum (n=1), P. catenulatum (n=3), P. pachycaule voucher (n=1), P. rhizo-oryzae (n=3), P. torulosum (n=4) e Pythium spp. (n=10). Zoósporos desses micro-organismos foram produzidos in vitro e inoculados por via subcutânea em coelhos, os quais foram avaliados durante 45 dias. Dentre os oomicetos testados, apenas P. insidiosum evidenciou patogenicidade, causando pitiose no modelo experimental, evidenciando que, em nossas condições, apenas esta espécie de Pythium é patógena para mamíferos.(AU)

One hundred and eighty-six water samples from swampy areas were collected in 13 municipalities of South, Central and West regions of Rio Grande do Sul, Brazil, in order to isolate and characterize Pythium species and assess their pathogenicity using rabbits as experimental model. Different Pythium species were isolated from 22 (11.8%) water samples, including P. insidiosum (n=1), P. catenulatum (n=3), P. pachycaule voucher (n=1), P. rhizo-oryzae (n=3), P. torulosum (n=4) e Pythium spp. (n=10). Zoospores of these microorganisms were produced in vitro and inoculated subcutaneously into rabbits, which were assessed over 45 days. Only P. insidiosum showed pathogenicity, causing pythiosis in the experimental model.(AU)

Presence of Mycobacterium leprae genotype 4 in environmental waters in Northeast Brazil

Abstract INTRODUCTION: This study quantified Mycobacterium leprae bacilli in environmental water samples from five municipalities in the State of Ceará by quantitative polymerase chain reaction (qPCR) and compared the identified genotypes with those obtained from leprosy patient biopsies. METHODS: We collected five replicas from each of the 30 selected reservoirs and skin lesion biopsies from 25 new leprosy cases treated at a reference center in Fortaleza, Ceará from 2010 to 2013. The 16S rRNA gene region of M. leprae was amplified by qPCR and a standard curve was created with the pIDTBlue 16SrRNAMlep plasmid. The Juazeiro do Norte water samples and the biopsies were genotyped (single nucleotide polymorphism [SNP] 1 to 4) and the SNP 4 genotypes were subtyped. RESULTS: Of the 149 water samples analyzed, 54.4% were positive for the M. leprae DNA. The M. leprae bacilli copy number ranged from 1.42 × 10 -1 to 1.44 × 10 + 2 . Most biopsies showed SNP type 4 (64%), while all samples from Juazeiro do Norte were SNP type 4, with subtype 4-N appearing at the highest frequency. CONCLUSIONS: We suggest that environmental waters containing M. leprae bacilli play an important role in disease transmission, justifying PGL-1 seropositivity in individuals living in areas where there is no reported case, and in leprosy cases individuals who report no previous contact with other case. Therefore, further investigation is needed to clarify disease transmission in this region and to explore the role of the environment. We also suggest that in this area surveillance for leprosy cases should be intensified.

Screening of freshwater fungi for decolorizing multiple synthetic dyes

Abstract The biodegradation of synthetic dyes by fungi is emerging as an effective and promising approach. In the present study, freshwater fungal strains isolated from submerged woods were screened for the decolorization of 7 synthetic dyes. Subsequently, 13 isolates with high decolorization capability were assessed in a liquid system; they belonged to 9 different fungal species. Several strains exhibited a highly effective decolorization of multiple types of dyes. New absorbance peaks appeared after the treatment with 3 fungal strains, which suggests that a biotransformation process occurred through fungal biodegradation. These results showed the unexploited and valuable capability of freshwater fungi for the treatment of dye-containing effluents. The ability of certain fungi to decolorize dyes is reported here for the first time.

Anaerobic microbiota: spatial-temporal changes in the sediment of a tropical coastal lagoon with ephemeral inlet in the Gulf of Mexico / Microbiota anaerobia: cambios espacio-temporales en el sedimento de una laguna costera tropical con boca efímera en el Golfo de México

Abstract:Sulfate-reducing bacteria (SRB) and methanogenic archaea (MA) share common niches in coastal sediments during the terminal phases of the anaerobic mineralization of organic matter. The purpose of this study was to analyze the spatial - temporal variation of SRB and MA in the sediments of a tropical coastal lagoon with ephemeral inlet (La Mancha, Veracruz, Gulf of Mexico) and its relationship with environmental changes. A total of 24 sediment samples were collected during the dry (April, May), rainy (July, September) and Northern (November, February) seasons in the period 2013-2014. Microbiological analyses included the quantification of the viable SRB and MA with different substrates, as well as mineralization experiments to determine the effect of sulfate on acetate oxidation. The analyzed environmental variables in the sediments included: temperature, pH, Eh, salinity, sulfates, H2S, volatile solids, carbohydrates, and granulometric characteristics. Major changes occurred between the dry and rainy seasons. During the dry season, sulfate-reducing abundance was significantly greater with lactate (8.3x105 - 1.2x107 cells / g) and propionate (1.8x105 - 6.6x106 cells / g) as substrates, while the MA that use methanol were dominant (4.2x105 - 9.1x106 cells / g). In contrast, during the rainy season, hydrogenophylic (2.6x105 - 8.3x106 cells/g) and acetoclastic (5.4x105-6.4x106 cells / g) MA increased significantly and SRB decreased in the analyzed substrates. An apparent competition for acetate was observed, with a greater oxidation in the media with sulfates in the dry season (0.06 mM acetate / g sediment / day), and a greater oxidation in the media without sulfates in the rainy season (0.02 mM acetate / g sediment / day). SRB and MA were present throughout the sediment column, however SRB dominated in the first centimeters of the sediment while MA were abundant in deeper layers. In conclusion, SRB and MA together played a role in the mineralization of organic matter in the sediments of La Mancha lagoon, with sulfate-reduction dominating in the dry season (closed inlet) and methanogenesis during the rainy season (open inlet). Changes in rainfall and river input in this lagoon significantly affect salinity and sulfate content, the main factors that regulate the dynamics of SRB and MA in the sediments. Rev. Biol. Trop. 64 (4): 1759-1770. Epub 2016 December 01.

Resumen:Las bacterias sulfatorreductoras (BSR) y las arqueas metanogénicas (AM) comparten nichos comunes en los sedimentos costeros durante las fases terminales de la mineralización anaerobia de la materia orgánica. El objetivo del presente estudio fue analizar la variación espacio-temporal de las BSR y AM en los sedimentos de una laguna costera tropical con una boca efímera (La Mancha, Veracruz, Golfo de México) y su relación con los cambios ambientales. Un total de 24 muestras de sedimentos fueron recolectadas en las temporadas de secas (abril, mayo), lluvias (julio, septiembre) y nortes (noviembre, febrero) en el periodo 2013 - 2014. Los análisis microbiológicos incluyeron la cuantificación de las BSR y AM viables con diferentes sustratos, así como experimentos de mineralización para determinar el efecto de los sulfatos en la oxidación del acetato. Las variables ambientales analizadas en los sedimentos incluyeron la temperatura, pH, Eh, salinidad, sulfatos, H2S, sólidos volátiles, carbohidratos y las características granulométricas. Los principales cambios se presentaron entre las estaciones de secas y lluvias. En la temporada de secas la abundancia sulfatorreductora fue significativamente superior con lactato (8.3x105 - 1.2x107 células / g) y propionato (1.8x105 - 6.6x106 células / g) como sustratos, mientras que las AM que emplean metanol dominaron (4.2x105 - 9.1x106 células / g). Por el contrario, en la época de lluvias aumentaron significativamente las AM hidrogenofílicas (2.6x105-8.3x106 células / g) y acetoclásticas (5.4x105-6.4x106 células / g), disminuyendo las BSR con los sustratos analizados. Se determinó una competencia aparente por el acetato. Su oxidación fue mayor en los medios con presencia de sulfatos en las temporadas de secas (0.06 mM acetato / g sedimento / día), mientras que aumentó en los medios sin sulfatos en la época de lluvias (0.02 mM acetato / g sedimento / día). Las BSR y las AM estuvieron presentes a lo largo de la columna sedimentaria; sin embargo, las BSR dominaron en los primeros centímetros del sedimento y las AM abundaron a mayor profundidad. En conclusión las BSR y AM participaron de manera conjunta en la mineralización de la materia orgánica en los sedimentos de la laguna de La Mancha, dominando la sulfatorreducción en la temporada de secas (boca cerrada) y la metanogénesis en la época de lluvias (boca abierta). Los cambios en el aporte fluvial y la precipitación en esta laguna afectaron significativamente la salinidad y el contenido de sulfatos, siendo los principales factores que regularon la dinámica de las BSR y AM en los sedimentos.

Cellulase activity and dissolved organic carbon release from lignocellulose macrophyte-derived in four trophic conditions

Abstract Considering the importance of lignocellulose macrophyte-derived for the energy flux in aquatic ecosystems and the nutrient concentrations as a function of force which influences the decomposition process, this study aims to relate the enzymatic activity and lignocellulose hydrolysis in different trophic statuses. Water samples and two macrophyte species were collected from the littoral zone of a subtropical Brazilian Reservoir. A lignocellulosic matrix was obtained using aqueous extraction of dried plant material (≈40 °C). Incubations for decomposition of the lignocellulosic matrix were prepared using lignocelluloses, inoculums and filtered water simulating different trophic statuses with the same N:P ratio. The particulate organic carbon and dissolved organic carbon (POC and DOC, respectively) were quantified, the cellulase enzymatic activity was measured by releasing reducing sugars and immobilized carbon was analyzed by filtration. During the cellulose degradation indicated by the cellulase activity, the dissolved organic carbon daily rate and enzyme activity increased. It was related to a fast hydrolysable fraction of cellulose that contributed to short-term carbon immobilization (ca. 10 days). After approximately 20 days, the dissolved organic carbon and enzyme activity were inversely correlated suggesting that the respiration of microorganisms was responsible for carbon mineralization. Cellulose was an important resource in low nutrient conditions (oligotrophic). However, the detritus quality played a major role in the lignocelluloses degradation (i.e., enzyme activity) and carbon release.

Characterization of the variable region in the class 1 integron of antimicrobial-resistant Escherichia coli isolated from surface water

Abstract Fecal bacteria are considered to be a potential reservoir of antimicrobial resistance genes in the aquatic environment and could horizontally transfer these genes to autochthonous bacteria when carried on transferable and/or mobile genetic elements. Such circulation of resistance genes constitutes a latent public health hazard. The aim of this study was to characterize the variable region of the class 1 integron and relate its genetic content to resistance patterns observed in antimicrobial-resistant Escherichia coli isolated from the surface waters of Patos Lagoon, Southern Brazil. Genetic diversity of the isolates and presence of the qacEΔ1 gene, which confers resistance to quaternary ammonium compounds, were also investigated. A total of 27 isolates were analyzed. The variable region harbored dfrA17, dfrA1 and dfrA12 genes, which confer resistance to trimethoprim, and aadA1, aadA5 and aadA22 genes that encode resistance to streptomycin/spectinomycin. Most of the isolates were considered resistant to quaternary ammonium compounds and all of them carried the qacE Δ1 gene at the 3′ conserved segment of the integron. ERIC-PCR analyses of E. coli isolates that presented the integrons showed great genetic diversity, indicating diverse sources of contamination in this environment. These results suggest that fecal bacteria with class 1 integrons in aquatic environments are potentially important reservoirs of antibiotic-resistance genes and may transfer these elements to other bacteria that are capable of infecting humans.

Microturbellarians (Platyhelminthes and Acoelomorpha) in Brazil: invisible organisms? / Microturbelários (Platyhelminthes e Acoelomorpha) no Brasil: organismos invisíveis?

Abstract Microturbellarians typically belong to the benthos and may occur in a wide variety of environments. They are abundant in freshwater and marine ecosystems and may occur in moist terrestrial habitats. However, turbellarians are seldom taken into account in studies of biodiversity. Most studies on Brazilian microturbellarians had taxonomical purposes and were done in the years 1940-1950. Thus, information on their occurrence and ecological aspects are dispersed throughout several papers. We intend here to summarize the biogeographical distribution and ecological aspects of microturbellarians recorded for Brazil, indicating the main gaps in their knowledge and possible actions to enhance studies on this group. There are 239 species of microturbellarians registered for Brazil, with records distributed in 12 states. However, just three states located in southern Brazil have records of 94% of microturbellarian species. Thus, knowledge on the systematics and geographical distribution of Brazilian microturbellarians clearly reflect the scientific activity over many years or decades in two states of southeastern and southern Brazil. Considering the scant information on this group in Brazil, which is also the situation of the Neotropical microturbellarians in general, some actions should be proposed. First, it would be necessary to sample in the diverse biomes, as well as in the various river and sea basins, based on standardized sampling protocols. Second, it would be necessary to encourage diverse research groups to include microturbellarians and/or turbellarians in general into biodiversity inventories and studies on community structure of invertebrates. Third, it is necessary to increase the number of research groups on microturbellarians, in order to augment the studies on their morphology, systematics, and ecology. Considering their abundance, species richness and ecological importance in aquatic environments, despite some peculiarities regarding their sampling, sorting and identification procedures, the challenge to study microturbellarians and enhance knowledge about them in Brazilian ecosystems should be faced.

Resumo Microturbelários são tipicamente bentônicos e podem ocorrer em uma ampla variedade de ambientes. São abundantes em ecossistemas marinhos e de água doce, podendo ocorrer em ambientes terrestres úmidos. Entretanto, turbelários raramente são considerados em estudos de diversidade. A maioria dos estudos sobre microturbelários brasileiros tiveram propósito taxonômico e foram realizados nos anos 1940-1950. Assim, informações sobre ocorrência e aspectos ecológicos estão dispersos em diversos artigos. O objetivo deste trabalho é sumarizar a distribuição biogeográfica e aspectos ecológicos dos microturbelários registrados para o Brasil, indicando as principais lacunas do conhecimento e possíveis ações para ampliar estudos sobre esse grupo. Há 239 espécies de microturbelários registradas no Brasil, com registros distribuídos em 12 estados. No entanto, 94% das espécies de microturbelários foram registradas em apenas três estados localizados no sul e sudeste do Brasil. Assim, o conhecimento sobre a sistemática e distribuição geográfica dos microturbelários claramente reflete as atividades científicas realizadas por muitos anos ou mesmo décadas em dois estados do sudeste e sul do Brasil. Considerando as escassas informações existentes sobre esse grupo no Brasil, assim como a situação dos microturbelários neotropicais em geral, algumas ações devem ser propostas. Primeiramente, é necessário realizar amostragens em diversos biomas, assim como nas várias bacias e regiões hidrográficas marinhas, baseadas em protocolos de amostragem padronizados. Em segundo lugar, faz-se necessário incentivar diversos grupos de pesquisa a incluir microturbelários e/ou turbelários em geral em inventários da biodiversidade e estudos de estrutura de comunidades de invertebrados. Em terceiro lugar, é necessário ampliar o número de grupos de pesquisa em microturbelários, para aumentar os estudos sobre sua morfologia, sistemática e ecologia. Considerando sua abundância, riqueza de espécies e importância ecológica em ambientes aquáticos, apesar de suas peculiaridades de amostragem, triagem e identificação, o desafio de estudar e ampliar o conhecimento sobre microturbelários em ecossistemas brasileiros deve ser enfrentado.

New host record of five Flavobacterium species associated with tropical fresh water farmed fishes from North India

Abstract Yellow pigmented, filamentous, Gram-negative bacteria belonging to genus Flavobacterium are commonly associated with infections in stressed fish. In this study, inter-species diversity of Flavobacterium was studied in apparently healthy freshwater farmed fishes. For this, ninety one yellow pigmented bacteria were isolated from skin and gill samples (n = 38) of three farmed fish species i.e. Labeo rohita, Catla catla and Cyprinus carpio. Among them, only twelve bacterial isolates (13.18%) were identified as Flavobacterium spp. on the basis of morphological, biochemical tests, partial 16S rDNA gene sequencing and phylogenetic analysis. On the basis of 16S rDNA gene sequencing, all the 12 isolates were 97.6-100% similar to six different formally described species of genus Flavobacterium. The 16S rDNA based phylogenetic analysis grouped these strains into six different clades. Of the 12 isolates, six strains (Fl9S1-6) grouped with F. suncheonense, two strains (Fl6I2, Fl6I3) with F. indicum and the rest four strains (Fl1A1, Fl2G1, Fl3H1 and Fl10T1) clustered with F. aquaticum, F. granuli, F. hercynium and F. terrae, respectively. None of these species except, F. hercynium were previously reported from fish. All the isolated Flavobacterium species possessed the ability of adhesion and biofilm formation to colonize the external surface of healthy fish. The present study is the first record of tropical freshwater farmed fishes as hosts to five environmentally associated species of the Flavobacterium.

The hyporheic zone and its functions: revision and research status in Neotropical regions / A zona hiporréica e as suas funções: revisão e estado da arte da pesquisa na região neotropical

AbstractThe hyporheic zone (HZ), as the connecting ecotone between surface- and groundwater, is functionally part of both fluvial and groundwater ecosystems. Its hydrological, chemical, biological and metabolic features are specific of this zone, not belonging truly neither to surface- nor to groundwater. Exchanges of water, nutrients, and organic matter occur in response to variations in discharge and bed topography and porosity. Dynamic gradients exist at all scales and vary temporally. Across all scales, the functional significance of the HZ relates to its activity and connection with the surface stream. The HZ is a relatively rich environment and almost all invertebrate groups have colonized this habitat. This fauna, so-called hyporheos, is composed of species typical from interstitial environment, and also of benthic epigean and phreatic species. The hyporheic microbiocenose consists in bacteria, archaea, protozoa and fungi. The HZ provides several ecosystem services, playing a pivotal role in mediating exchange processes, including both matter and energy, between surface and subterranean ecosystems, functioning as regulator of water flow, benthic invertebrates refuge and place of storage, source and transformation of organic matter. The hyporheic zone is one of the most threatened aquatic environments, being strongly influenced by human activities, and the least protected by legislation worldwide. Its maintenance and conservation is compelling in order to preserve the ecological interconnectivity among the three spatial dimensions of the aquatic environment. Although several researchers addressed the importance of the hyporheic zone early, and most contemporary stream ecosystem models explicitly include it, very little is known about the HZ of Neotropical regions. From a biological standpoint, hyporheos fauna in Neotropical regions are still largely underestimated. This review focuses on a brief presentation of the hyporheic zone and its functions and significance as an ecotone. We also highlighted the key aspects considering also the current status of research in Neotropical regions.

ResumoA zona hiporréica, como ecótono de ligação entre a superfície e as águas subterrâneas, é parte funcional seja dos ecossistemas fluviais seja das águas subterrâneas. As características hidrológicas, as características químicas, biológicas e metabólicas são específicas desta zona, não pertencendo verdadeiramente nem a superfície nem às águas subterrâneas. Trocas de água, nutrientes e matéria orgânica ocorrem em resposta a variações na descarga, topografia do álveo e porosidade. Gradientes dinâmicos existem em todas as escalas e variam temporalmente. Em todas as escalas, o significado funcional da zona hyporheic relaciona-se com a sua conexão e atividades com a água superficial. O HZ é um ambiente relativamente rico e quase todos os grupos de invertebrados colonizaram este habitat. Esta fauna, chamada hyporheos, é composta por espécies típicas do ambiente intersticial, e também de espécies bentônicas epígeas e freáticas. A microbiocenose consiste em bactérias, arqueobactérias, fungos e protozoários. O HZ fornece vários serviços para o ecossistema, desempenhando um papel fundamental na mediação de processos de troca, incluindo seja a matéria, seja a energia, entre os ecossistemas superfíciais e os subterrâneos, funcionando como regulador do fluxo de água, de refúgio para invertebrados bentônicos e local de armazenagem, fonte e transformação de matéria orgânica. A zona hyporheic é um dos ambientes aquáticos mais ameaçados, sendo fortemente influenciado pelas atividades humanas, e um dos menos protegidos pela legislação em todo o mundo. A sua manutenção e conservação é necessaria para preservar a interconectividade ecológica entre as três dimensões espaciais do ambiente aquático. Apesar de vários pesquisadores aborem a importância da zona hyporheic a tempo, e a maioria dos modelos de ecossistemas atualmente incluí-lo de forma explicita, muito pouco se sabe sobre o HZ das regiões neotropicais. Do ponto de vista biológico, a fauna hiporréica das regiões neotropicais é ainda largamente subestimada. Esta revisão visa apresentar de forma resumida a zona hiporréica, suas funções e importância como ecótono. Também visa destacar os aspectos principais considerando também o estado actual da investigação em regiões neotropicais.

Cyanobacteria bloom: selective filter for zooplankton? / Floração de cianobactéria: filtro seletivo para zooplâncton?

The Ibirité reservoir is an urban and eutrophic environment, with regular occurrences of cyanobacteria blooms. The reservoir is warm monomict and remains stratified most of the year, circulating in the dry season (winter). During the hydrological cycle of October/07 to October/08 there were four scenarios with different environmental conditions, which influenced the structure of the zooplankton community, as confirmed in a previous study. Changes in the zooplankton community structure between the scenarios were studied, aiming at analyzing the stability and persistence of this community. The Spearman’s coefficient of correlation was used to measure the stability; the persistence was evaluated through a cluster analysis and changes in community composition were estimated by the "temporal" β diversity index. Considering the distribution patterns of abundance, the community was stable only in the transition between scenarios 1 and 2 (n = 30, r = 0.71, p = 0.00001), when there were no cyanobacteria blooms. The persistence of zooplankton between the scenarios was low, showing a distinct species composition for each scenario. The highest variations in species composition, observed by the values of temporal β diversity index, were the transitions between scenarios 3-0 (1.45) and 0-1 (1.05), and the lowest variations occurred in the transition between scenarios 1-2 (0.57). The results suggest that the cyanobacteria blooms at Ibirité reservoir are be acting as "selective filters", and are, thus, disturbances with sufficient ability to change the structure of the zooplankton community. Keywords Urban Reservoir, zooplankton, bloom of cyanobacteria, stability, persistence. .

O reservatório de Ibirité é um ambiente urbano, eutrófico com periódicas ocorrências de florações de cianobactérias. O padrão de mistura deste reservatório é monomítico quente, permanecendo estratificado a maior parte do ano, circulando na estação seca (inverno). Durante o ciclo hidrológico de outubro/07 a outubro/08, ocorreram quatro cenários com condições ambientais distintas, que influenciaram na estruturação da comunidade zooplanctônica, conforme observado em estudo prévio. As mudanças na estrutura da comunidade zooplanctônica entre os cenários foram estudadas objetivando analisar a estabilidade e persistência desta comunidade. O coeficiente de correlação de Spearman foi utilizado para medir a estabilidade; a persistência foi considerada por meio de uma análise de agrupamento; e as mudanças na composição da comunidade foram estimadas através do índice de diversidade β temporal. A comunidade foi estável, considerando os padrões de distribuição da abundância, somente na transição entre os cenários 1 e 2 (n=30; r= 0,71; p=0,00001), nos quais não foram observadas florações de cianobactérias. A persistência do zooplâncton entre os cenários foi baixa, apresentando composições de espécies distintas. As maiores variações na composição das espécies, estimadas através dos valores de diversidade β temporal, foram nas transições entre os cenários 3-0 (1,45) e 0-1 (1,05), e o menor valor na transição entre os cenários 1-2 (0,57). Os resultados do presente trabalho sugerem que as florações de cianobactérias no reservatório de Ibirité estariam funcionando como “filtros seletivos”, ou seja, distúrbios com capacidade suficiente para alterar a estrutura da comunidade zooplanctônica.

Molecular identification of the occurrence of magnetotactic bacteria in fresh water sediments (Czech Republic)

Magnetotactic bacteria (MTB) are of considerable interest because of their importance in the manufacture of various bioinspired materials. In order to find out the status of magnetotactic bacteria at three different sediment in Czech Republic, samples collected from both standing and running freshwaters were subjected to molecular diversity analysis by using 16S rRNA gene approach. Total community DNA from sediment sample was isolated and used for PCR, cloning and sequence analysis. Of the 24 analyzed sequences, six clones are closely related to Magnetobacterium sp. affiliated with the Nitrospira phylum which showed the dominance of Magnetobacterium phylotypes in the sample. This study will provide useful insight about the community structure of MTB in this particular geographical region. However more detailed and specific studies are warranted in order to properly assess the community structure of MTB's in fresh water sediments.

The importance of protozoan bacterivory in a subtropical environment (Lobo-Broa Reservoir, SP, Brazil) / A importância da bacterivoria por protozoários em um ambiente subtropical (Reservatório do Lobo-Broa, SP, Brasil)

This study evaluated the importance of heterotrophic nanoflagellates (HNF) and ciliates bacterivory in a mesotrophic subtropical environment (Lobo-Broa Reservoir, Brazil) by the quantification of their ingestion rates. The in situ experiments using fluorescently labelled bacteria (FLB) were carried out bimonthly over one year (three surveys in the dry season and three in the rainy one) at the sub-surface of two sampling points that have different trophic degrees. The ingestion rates for both ciliates and HNF were higher in the meso-eutrophic region (point 2) due to the higher water temperatures, which accelerate the metabolism of protozoans and the higher bacteria densities. Concerning total protozoan bacterivory, the HNF had the greatest grazing impact on bacterial community, especially the HNF <5µm. The data showed that HNF grazing, in addition to regulating the bacteria abundance, also induced changes to the bacterial community structure, such as increasing size and numbers of bacterial filaments. The ciliates were also important to the system bacterivory, especially in point 2, where there were high densities and ingestion rates. The protozoan bottom-up control was more important in the dry season and the top-down control was more important in the rainy season, so, these two forces are equally important to the bacterial abundance regulation in this reservoir in an annual basis.

Este estudo avaliou a importância da bacterivoria por nanoflagelados heterotróficos (NFH) e ciliados em um ambiente subtropical mesotrófico (Reservatório do Lobo-Broa, Brasil) pela quantificação de suas taxas de ingestão. Os experimentos in situ utilizando bactérias marcadas por fluorescência (FLB) foram realizados bimestralmente durante um ano (três estudos no período seco e três no período chuvoso) na superfície de dois pontos de coleta que apresentam graus de trofia diferentes. As taxas de ingestão, tanto para os ciliados quanto para os NHF, foram maiores no ponto 2 (P2) devido provavelmente as maiores temperaturas da água, que aceleram o metabolismo dos protozoários, e as maiores densidades de bactérias. Em relação à bacterivoria total por protozoários, as populações de NFH causaram maior impacto de predação sobre a comunidade bacteriana do reservatório, principalmente os NFH menores que 5µm. Os dados indicaram que a predação por NFH, além de regular a abundância de bactérias, provavelmente também induziu modificações na estrutura da comunidade bacteriana, como o aumento do tamanho e da quantidade de filamentos bacterianos. Os ciliados também contribuíram significativamente para a bacterivoria no sistema, principalmente no P2, onde apresentaram altas densidades e taxas de ingestão. O controle bottom-up teve maior importância no período seco e o controle top-down maior importância no período chuvoso, assim, estas duas forças são igualmente importantes na regulação da abundância bacteriana neste reservatório em uma base anual.

Técnicas moleculares para la detección de cianobacterias en los embalses Riogrande II y La Fe, Colombia / Molecular techniques for cyanobacteria detection at Riogrande II and La Fe water reservoirs, Colombia

In lentic water bodies as reservoirs occur eutrophication processes, originated mainly from human activities (i.e. agriculture, animal exploitation). This influx of nutrients in aquatic ecosystems could promote blooms of potentially toxic cyanobacteria. The purpose of this work is to detect the presence of cyanobacteria strains in water samples, using molecular techniques to help in preventive management of reservoirs dedicated to water purification. We used two molecular techniques to detect genes implied with the synthesis of hepatotoxic microcystins from potentially toxic cyanobacteria strains, and to evaluate the molecular diversity of cyanobacteria in water samples from two high-mountain reservoirs used for purification of drinking water for the metropolitan area of Medellin, Colombia. Between 2010-2011 collections of 12 water samples were taken and DNA extraction together with PCR and DGGE analyses where carried out. We amplified 22 sequences between 250-300bp of the genes mcyA and mcyE, and these sequences were related with several strains and cyanobacteria genera accessions from NCBI-GenBank databases. Moreover, sequence amplifications of the 16S small ribosomal RNA subunit - 16S rRNA- between 400-800bp were also performed in order to use them for the DGGE technique. The amplification products of DGGE were set in polyacrilamide gel with posterior denaturing electrophoresis, and the scanned images of the gel bands were analysed with the software GelCompar II. For Riogrande II and La Fe reservoirs we found 35 and 30 different DGGE bands, respectively, as a measurement of molecular diversity in these artificial ecosystems. Here, we demonstrated the utility of two molecular techniques for the detection of genes associated with toxicity and molecular diversity of cyanobacteria in reservoirs destined for drinking water in urban centers. We recommend strongly following with periodically molecular biology studies in these ecosystems combined with limnological and ecological data, as new tools for management of plants of water purification and for the prevention of potentially public health concerns. Rev. Biol. Trop. 62 (1): 403-419. Epub 2014 March 01.

En embalses, la eutrofización es consecuencia de procesos naturales y de actividades humanas, lo cual puede facilitar la aparición de afloramientos de cianobacterias potencialmente tóxicas. En este trabajo, se utilizaron dos técnicas moleculares en la detección de genes presentes en cepas potencialmente tóxicas de cianobacterias y evaluaciones de diversidad molecular de cianobacterias en los embalses para potabilización de agua, Riogrande II y La Fe, Colombia. Entre 2010-2011, se tomaron 12 muestras de agua en ambos embalses y se realizaron extracciones de ADN para un análisis de marcadores moleculares mediante PCR y DGGE. Se amplificaron secuencias entre 250-300pb de los genes mcyA y mcyE, implicados en la toxicidad de cepas de cianobacterias. Asimismo, se amplificaron secuencias de la región 16S del ARN ribosomal (422pb), para la técnica DGGE. Se corrieron geles de poliacrilamida en gradientes de desnaturalización, se realizó agrupamiento genético (UPGMA), y se separaron por patrones de bandeo las muestras de cada embalse evaluado. Se demuestra la utilidad de las técnicas moleculares en estudios relacionados con la búsqueda de genes asociados con toxicidad y diversidad molecular de cianobacterias en muestras de agua provenientes de embalses de agua con fines de potabilización para centros urbanos.

The climatic rhythm and blooms of cyanobacteria in a tropical reservoir in São Paulo, Brazil / O ritmo climático e as florações de cianobactérias em um reservatório tropical em São Paulo, Brasil

The present study sought to develop a methodology to analyse water quality based on the concepts and methods of climate and climatology. Accordingly, we attempted to relate hydro- and limnometeorological techniques and methodologies to a rhythmic analysis technique developed within the context of the Brazilian geographical climatology. Our goal was to assess and analyse cyanobacterial blooms, the main index of water quality for the reservoirs of the “Alto Tietê” Basin and, consequently, the Metropolitan Region of São Paulo, an area of high environmental complexity due to its high degree of development and high population density. The meteorological data used were collected by the Institute of Astronomy, Geophysics and Atmospheric Sciences at the University of São Paulo meteorological station, and the limnological data were collected through the Hydrological Monitoring System implemented by SABESP in the Billings and Guarapiranga Reservoirs and the laboratory of the same entity. The rhythmic and integrated analysis showed that the process of cyanobacterial blooms is dependent on a combination of meteorological factors as temperature and wind intensity that may disrupt the stability of the reservoir, providing the conditions necessary for the development of cyanobacteria during the stabilisation process. The pace of the Atlantic Polar Front Entrance during the winter in São Paulo is a limiting factor for the growth of cyanobacteria because of their high frequency, thus maintaining the balance of the reservoir throughout this period. The weather types those could cause a instability in the water column were: Cold Front entrance (66.67%), conflict between masses (22.22%) and the Tropical Instability Line (11.11%). The possibility for prevention and forecasting periods advise when these reservoirs should not be used, mainly with regard to recreational activities.

O presente trabalho buscou desenvolver uma metodologia para análise da qualidade da água com base em conceitos e métodos do clima e da climatologia respectivamente. Dessa forma, buscou-se relacionar técnicas e metodologias hidrometeorológicas e limno-meteorológicas com a técnica de análise rítmica desenvolvida dentro da climatologia geográfica brasileira para avaliar e analisar as florações de cianobactérias, principal índice de qualidade de água encontrado nos reservatórios da Bacia do Alto Tietê e consequentemente da Região Metropolitana de São Paulo, área de alta complexidade ambiental devido ao seu alto grau de desenvolvimento e a sua alta densidade populacional. Os dados meteorológicos utilizados para o desenvolvimento do trabalho foram coletados a partir da estação meteorológica do Instituto de Astronomia, Geofísica e Ciências Atmosféricas da Universidade de São Paulo e os dados limnológicos foram coletados por meio do Sistema de Monitoramento Hidrológico implantado pela SABESP nos reservatórios de Billings e Guarapiranga assim como os dados laboratoriais da mesma entidade. A análise rítmica e integrada de todos os dados mostrou que o processo de floração de cianobactérias é dependente não apenas de um fator específico, mas de uma combinação de fatores meteorológicos como temperatura e intensidade do vento que podem quebrar a estabilidade do reservatório e proporcionar durante a estabilização as condições necessárias para o desenvolvimento das cianobactérias. Também foi mostrado como o ritmo de entrada de Frentes Polares Atlânticas, durante o inverno paulistano, é um fator que limita o crescimento das cianobactérias devido à sua alta frequência, mantendo o reservatório desequilibrado por todo o período. Os tipos de tempo que podem causar instabilidade na coluna da água foram identificados com: entrada de Frentes Frias (66,67%), conflitos entre massas (22,22%) e Linha de Instabilidade (11,11%). Mostra-se assim a importância do trabalho para a prevenção e previsão de períodos inapropriados para o uso desses reservatórios principalmente para atividade de recreação.

The influence of fish culture in floating net cages on microbial indicators of water quality / Influencia da piscicultura em tanques-rede sobre os microrganismos indicadores da qualidade de agua

This work was carried out to analyse the microbiological parameters of the water quality of a reservoir used for the irrigation and culture of tilapia (Oreochromis niloticus) in floating net cages. The physico-chemical parameters, counts of mesophilic total aerobic bacteria, total and thermotolerant coliforms and fecal streptococci, and the presence of Escherichia coli in samples of water collected in three sites of the reservoir (pre-culture site, culture site, post-culture site) were analysed. The levels of ammonia (0.047-0.059 mg/L), nitrite (0.001-0.021 mg/L) and total phosphorus (0.050-0.355 mg/L) in the water did not show significant differences (p > 0.05) between sampling sites. The levels of total bacteria in the water varied between 1.3 x 104 and 67.3 x 104 CFU/100 mL. The MPN values of thermotolerant coliforms (< 930 MPN/100 mL) were within values recommended for water used for fish culture and/or irrigation. The presence of E. coli and fecal streptococci were verified in 48% and 56% of analysed samples, respectively. The site with floating net cages showed more samples contaminated with E. coli and fecal streptococci than other sampling points.

Este trabalho foi realizado para analisar os parâmetros microbiológicos da qualidade da água de um reservatório destinado a irrigação e à cultivo de tilapia do Nilo (Oreochromis niloticus) em tanques redes. Foi realizada análise de parâmetros físico-químicos, quantificação de bactérias totais aeróbias mesófilas, coliformes totais e termotolerantes, estreptococos fecais e a detecção de E. coli em amostras de água coletada em três pontos distintos do açude (pré-cultivo, cultivo, pós-cultivo). Os níveis de amônia (0,047-0,059 mg/L), nitrito (0,001-0,021 mg/L) e fósforo total (0,050-0,355 mg/L) na água não apresentaram diferenças significativas (p > 0.05) entre os pontos de coleta. As contagens de bactérias aeróbias mesófilas variaram entre 1,3 x 104 e 67,3 x 104 UFC/100 mL nas amostras de água. O NMP de coliformes termotolerantes (< 930 NMP/100 mL) se apresentou dentro do limite permitido para águas doces destinadas a piscicultura e/ou irrigação. A presença de E. coli foi constatada em 48% e de estreptococos fecais em 56% das amostras de água analisadas. O local de cultivo de tilápia em tanques-rede apresentou maior número de amostras contaminadas com E. coli e estreptococos fecais que outros pontos de amostragem.

Change rules of quantity and species of diatoms in Hunhe River in Shenyang / 法医学杂志

OBJECTION@#To observe the change rules of quantity and species of diatoms in Hunhe River in Shenyang and to provide technology and scientific evidence for drowning identification and the location of drowning in forensic investigation.@*METHODS@#In 2011, different locations for collecting water samples were chosen in Hunhe River in Shenyang. Water samples were collected and variation of quantity and species of diatoms were observed every month. And variation of dominant species of diatoms was observed every week.@*RESULTS@#The quantity, species and dominant species of diatoms in Hunhe River in Shenyang varied with different time and locations. The quantity and species of diatoms were lowest from December to February and gradually increased, reaching peak in May and second peak in October, and then gradually decreased. The dominant species of diatoms varied significantly adjacent two weeks at same location from April to November, but had little changes at different locations in same week from July to August.@*CONCLUSION@#The change rules of quantity and species of diatoms are complicated and affected by various factors such as environment and hydrology. The change rules of species and quantity of diatoms should be considered in forensic investigation of drowning identification and the location of drowning.

Susceptibilidad antimicrobiana de aislamientos de Escherichia coli procedentes de ecosistemas dulceacuícolas / Antimicrobial susceptibility of Escherichia coli isolates from river water ecosystems

Introducción: la resistencia antimicrobiana constituye uno de los mayores problemas que afronta la salud pública mundial. La aparición de cepas resistentes no solo de origen clínico sino también ambiental agrava la situación. Entre los microorganismos que presentan esta característica se destaca la especie bacteriana Escherichia coli debido a su doble papel como indicador de contaminación fecal y como patógeno. Objetivos: aislar e identificar hasta especie aislamientos de Escherichia coli a partir de muestras de agua procedentes de ríos contaminados de La Habana y determinar la susceptibilidad antimicrobiana in vitro de estos aislados. Métodos: se estudiaron 113 aislamientos de bacterias coliformes aislados de 10 estaciones de muestreo ubicadas en la zona urbana de los ríos capitalinos Almendares, Quibú y Luyanó en el período comprendido de febrero de 2008 hasta junio de 2010. La identificación de los aislados, la determinación de la susceptibilidad antimicrobiana y la búsqueda de b-lactamasas de espectro extendido se realizó mediante el método automatizado VITEK. Resultados: se identificaron 113 cepas ambientales de Escherichia coli. Se demostró que 23 % de los aislamientos resultaron resistentes al menos a uno de los antimicrobianos evaluados. Los mayores porcentajes de resistencia se observaron frente a ampicilina, sulfametoxazol-trimetropin y ciprofloxacina. Conclusiones: la presencia de aislados de E. coli con multirresistencia antimicrobiana en estos ríos indica claramente el riesgo biológico que implica el uso de sus aguas.

Introduction: antimicrobial resistance is one of the biggest problems facing global public health. The emergence of resistant clinical and environmental strains worsens the situation. Among the microorganisms with antimicrobial resistance, Escherichia coli species stands out due to its dual role as fecal contamination indicator and pathogen. Objectives: to isolate and identify Escherichia coli isolates from water samples from polluted rivers located in La Habana, and to determine their antimicrobial in vitro susceptibility. Methods: one hundred thirteen isolates of coliform bacteria isolated from 10 sampling stations in the capital´s urban areas near Almendares, Quibú and Luyanó rivers were studied in the period of February 2008 to June 2010. The identification of isolates, the determination of antimicrobial susceptibility and the search for extended-spectrum b-lactamase were all performed using VITEK automated method. Results: one hundred thirteen environmental strains of Escherichia coli were identified. It showed that 23 % of the isolates were resistant to at least one of the tested antimicrobials. The highest percentages of resistance were observed to ampicilline, sulfamethoxazole-trimethoprim and ciprofloxacin. Conclusions: the presence of E. coli isolates with multiple antimicrobial resistances in these rivers clearly indicates the biological risk involving the use of their waters.

Seasonal dynamics of cyanobacteria in a eutrophic reservoir (Arcoverde) in a semi-arid region of Brazil / Dinâmica sazonal de cianobactérias em um reservatório eutrófico (Arcoverde) no semiárido brasileiro

Environmental conditions favor the predominance of dense populations of cyanobacteria in reservoirs in northeastern Brazil. The aim of this study was to understand cyanobacterial population dynamics in the rainy and dry seasons at two depths in the Arcoverde reservoir. Microalgae and cyanobacteria samples were collected during 24 hours with intervals of 4 hours (nycthemeral) at sub-surface and 10 m using a van Dorn bottle and a determined biomass. Physical and chemical variables were obtained and the data were analyzed using the principal component analysis (PCA). No nycthemeral variations in the taxonomic composition or distribution of the populations of cyanobacteria were found between the different times of day in either the rainy or dry season. In both seasons, the greatest biomass of the phytoplankton community was made up of cyanobacteria at two depths and all times of the day. Cylindrospermopsis raciborskii (Woloszynska) Seenayya et Subba Raju was dominant at all times of the day on both the surface and at the bottom. In the rainy season, the differences in cyanobacterial biomass between the surface and bottom were less significant than in the dry season. The differences in cyanobacterial biomass between surface and bottom were less pronounced than those found in the dry season. We concluded that a) physical variables better explain the alterations of species in the phytoplankton community in an environment dominated by cyanobacteria throughout the year; b) seasonal climatic factors associated to periods of stratification and de-stratification are important for alterations in the community and variations in biomass and, c) the turbidity caused by rainfall favored the emergence and establishment of other cyanobacteria, especially Planktothrix agardhii (Gomont) Anagnostidis & Komárek.

As condições ambientais favorecem a predominância de populações densas de cianobactérias no nordeste do Brasil. O objetivo deste estudo foi entender a dinâmica das populações de cianobactérias nas estações seca e chuvosa, em duas profundidades, no reservatório de Arcoverde. Amostras de microalgas e cianobactérias foram coletadas durante 24 horas em intervalos de quatro horas (nictemerais) na subsuperfície e em 10 m, com uma garrafa de van Dorn, para a determinação da biomassa. Variáveis físicas e químicas foram obtidas, e os dados investigados por meio da Análise de Componentes Principais (ACP). Não foram encontradas variações nictemerais na composição taxonômica ou na distribuição das populações de cianobactérias entre os diferentes horários do dia, bem como entre as estações seca e chuvosa. Em ambas as estações, horários do dia e profundidades amostrais, a maior biomassa da comunidade fitoplanctônica foi constituída por cianobactérias. Cylindrospermopsis raciborskii (Woloszynska) Seenayya et Subba Raju foi dominante em todos os horários do dia, tanto na superfície como no fundo. Na estação chuvosa, as diferenças de biomassa de cianobactérias entre superfície e fundo foram menos pronunciadas do que na estação seca. Concluímos que: a) variáveis físicas explicam melhor as alterações das espécies na comunidade fitoplanctônica em um ambiente dominado por cianobactérias durante todo o ano; b) fatores climáticos sazonais associados a períodos de estratificação e desestratificação são importantes para as alterações na comunidade e as variações na biomassa, e c) a turbidez causada pela chuva favoreceu o aparecimento e o estabelecimento de outras cianobactérias, especialmente Planktothrix agardhii (Gomont) Anagnostidis & Komárek.