Monilinema gen. nov., a homocytous genus (Cyanobacteria, Leptolyngbyaceae) from saline-alkaline lakes of Pantanal wetlands, Brazil.

Cyanobacteria morphotypes with simple morphology, mainly thin filamentous homocytous strains, comprise a taxonomically complex group and represent a challenge in systematic studies. However, the polyphasic approach applied nowadays to investigate the cyanobacterial diversity has emerged as a powerful tool to undercover cryptic taxa and to set up a more natural classification system. Yet, studies exploring the cultured diversity of cyanobacteria from extreme tropical environments have paved the way to the discovery of new cyanobacteria taxa, enabling the description of Pantanalinema, Alkalinema, and Cephalothrix as novel genera from saline-alkaline lakes in Pantanal wetlands (Brazil). The present study investigated a set of cyanobacterial strains resembling Leptolyngbya morphotypes by means of its morphology, 16S ribosomal RNA (16S rRNA) phylogeny, and ITS secondary structures. Based on the shape and structure of their trichomes, the low levels of 16S rRNA identity coupled with the distinct phylogenetic position with regard to well establish genera, and their divergent ecological feature, these strains must be accommodated into a novel genus, named as Monilinema gen. nov., described under the provisions of the International Code of Nomenclature for algae, fungi, and plants. Accordingly, based on the singularity of the D1-D1' and V3 helixes, these strains should be considered as a single species, Monilinema alkalinum.

Neolyngbya gen. nov. (Cyanobacteria, Oscillatoriaceae): A new filamentous benthic marine taxon widely distributed along the Brazilian coast.

Brazil has an extensive and environmentally diverse coastline, which favors the occurrence of numerous cyanobacterial morpho- and ecotypes. Nevertheless, this coastline is still poorly studied and its diversity is underestimated. Considering the family Oscillatoriaceae, Lyngbya deserves special attention. It includes many clades which are phylogenetically non-related but morphologically similar. Such clades occur in marine and freshwater environments and are traditionally treated as a single genus. In the current study, we sampled both mediolittoral and estuarine zones along the Brazilian coast. Based on a polyphasic characterization, we described a new genus of marine filamentous cyanobacteria: Neolyngbya. It includes six new species sampled in Brazil, which are described in this study (N. maris-brasilis, N. granulosa, N. irregularis, N. nodulosa, N. arenicola and N. tenuis). Additionally, the characterization included a Neolyngbya sp. from Japan in the clade, but only based on molecular data. All species presented irregular arrangement of thylakoids as described for Oscillatoriaceae. The new genus shares morphological characteristics with species in different clades of the Lyngbya complex. The ultrastructural analyses of Neolyngbya, however, showed numerous gas vesicles, especially in the interthylakoid space; such feature is not observed in benthic Lyngbya species. Neolyngbya formed a well-supported clade (16S rRNA phylogeny), however distantly related to L. aestuarii and L. confervoides, both marine species clusters. The Limnoraphis clade is in a sister relationship to the Neolyngbya clade, however the former occurs in freshwater plankton. Secondary structures of 16S-23S rRNA ITS sequences were congruent with the phylogeny. The polyphasic characterization was helpful to clarify the diversity and ecological aspects of benthic filamentous cyanobacteria and the evolutionary history of the group. This favors a better understanding of inter and infrageneric taxa. The number of novel taxa described in this study emphasizes the importance of conducting additional floristic surveys, mainly in underexplored marine environments, to reveal the real cyanobacterial biodiversity in these areas.

Amazoninema gen. nov., (Synechococcales, Pseudanabaenaceae) a novel cyanobacteria genus from Brazilian Amazonian rivers.

The genus Leptolyngbya includes morphotypes with thin cells and simple morphology, and is one of the most common cyanobacterial genera found in a wide range of environments. In many cases, however, the morphotypes assigned to this genus do not share a common ancestor based on 16S rRNA gene phylogeny, which has led to the description of novel genera, such as Nodosilinea, Oculatella, Pantanalinema, Alkalinema, Thermoleptolyngbya, Onodrimia, Timaviella and Toxifilum. Thus, four novel isolates, with a comparable morphology to Leptolyngbya, were recovered from the Amazon and Solimões rivers. The novel 16S rRNA gene sequences obtained from these strains were placed together as a new and distinct phylogenetic lineage that is more closely related to the clusters embracing the genera Nodosilinea, Haloleptolyngbya and Halomicronema than to the genus Leptolyngbya. Additionally, these novel 16S rRNA gene sequences showed similarity values lower than 95 % compared with those from the most phylogenetic related groups and/or established genera. Altogether, these results supported the erection of a novel genus, named Amazoninema, to accommodate the novel isolates. Likewise, a comparison of their 16S rRNA gene sequences revealed similarities higher than 99.8 %, indicating that they belong to a single species, which was corroborated by analysing their 16S-23S internal transcribed spacer regions and unique Box-B helix pattern. Few studies have been undertaken to uncover the cultured diversity of cyanobacteria from Amazonia, and to our knowledge, this is the first cyanobacteria genus erected, considering morphotypes isolated exclusively from Brazilian Amazonian rivers.

Halotia gen. nov., a phylogenetically and physiologically coherent cyanobacterial genus isolated from marine coastal environments.

Nostoc is a common and well-studied genus of cyanobacteria and, according to molecular phylogeny, is a polyphyletic group. Therefore, revisions of this genus are urged in an attempt to clarify its taxonomy. Novel strains isolated from underexplored environments and assigned morphologically to the genus Nostoc are not genetically related to the 'true Nostoc' group. In this study, four strains isolated from biofilms collected in Antarctica and five strains originated from Brazilian mangroves were evaluated. Despite their morphological similarities to other morphotypes of Nostoc, these nine strains differed from other morphotypes in ecological, physiological and genetic aspects. Based on the phylogeny of the 16S rRNA gene, the Antarctic sequences were grouped together with the sequences of the Brazilian mangrove isolates and Nostoc sp. Mollenhauer 1 : 1-067 in a well-supported cluster (74 % bootstrap value, maximum-likelihood). This novel cluster was separated phylogenetically from the 'true Nostoc' clade and from the clades of the morphologically similar genera Mojavia and Desmonostoc. The 16S rRNA gene sequences generated in this study exhibited 96 % similarity to sequences from the nostocacean genera mentioned above. Physiologically, these nine strains showed the capacity to grow in a salinity range of 1-10 % NaCl, indicating their tolerance of saline conditions. These results provide support for the description of a new genus, named Halotia gen. nov., which is related morphologically to the genera Nostoc, Mojavia and Desmonostoc. Within this new genus, three novel species were recognized and described based on morphology and internal transcribed spacer secondary structures: Halotia branconii sp. nov., Halotia longispora sp. nov. and Halotia wernerae sp. nov., under the provisions of the International Code of Nomenclature for Algae, Fungi and Plants.

Pantanalinema gen. nov. and Alkalinema gen. nov.: novel pseudanabaenacean genera (Cyanobacteria) isolated from saline-alkaline lakes.

The genus Leptolyngbya Anagnostidis & Komárek (1988) was described from a set of strains identified as 'LPP-group B'. The morphology within this group is not particularly informative and underestimates the group's genetic diversity. In the present study, two new pseudanabaenacean genera related to Leptolyngbya morphotypes, Pantanalinema gen. nov. and Alkalinema gen. nov., are described under the provisions of the International Code of Nomenclature for Algae, Fungi and Plants, based on a polyphasic approach. Pantanalinema gen. nov. (type species Pantanalinema rosaneae sp. nov.) has sheaths and trichomes with slight gliding motility, which distinguish this genus from Alkalinema gen. nov. (type species Alkalinema pantanalense sp. nov.), which possesses trichomes arranged in an ornate (interwoven) pattern. 16S rRNA gene sequences of strains of Pantanalinema and Alkalinema exhibited low identity to each other (≤91.6 %) and to other sequences from known pseudanabaenacean genera (≤94.3 and 93.7 %, respectively). In a phylogenetic reconstruction, six sequences from strains of Pantanalinema and four from strains of Alkalinema formed two separate and robust clades (99 % bootstrap value), with the genera Oculatella and Phormidesmis, respectively, as the closest related groups. 16S-23S rRNA intergenic spacer sequences and secondary structures of strains of Pantanalinema and Alkalinema did not correspond to any previous descriptions. The strains of Pantanalinema and Alkalinema were able to survive and produce biomass at a range of pH (pH 4-11) and were also able to alter the culture medium to pH values ranging from pH 8.4 to 9.9. These data indicate that cyanobacterial communities in underexplored environments, such as the Pantanal wetlands, are promising sources of novel taxa.

Cephalothrix gen. nov. (Cyanobacteria): towards an intraspecific phylogenetic evaluation by multilocus analyses.

For more than a decade, the taxonomy of the Phormidiaceae has been problematic, since morphologically similar organisms represent phylogenetically distinct entities. Based on 16S rRNA gene sequence analyses, the polyphyletic genus Phormidium and other gas-vacuolated oscillatorioids appear scattered throughout the cyanobacterial tree of life. Recently, several studies have focused on understanding the oscillatorioid taxa at the generic level. At the specific level, few studies have characterized cyanobacterial strains using combined datasets (morphology, ultrastructure and molecular multilocus analyses). Using a multifaceted approach, we propose a new, well-defined genus, Cephalothrix gen. nov., by analysing seven filamentous strains that are morphologically 'intermediate' between gas-vacuolated taxa and Phormidium. Furthermore, we characterize two novel species: Cephalothrix komarekiana sp. nov. (strains CCIBt 3277, CCIBt 3279, CCIBt 3523, CCALA 155, SAG 75.79 and UTEX 1580) and Cephalothrix lacustris sp. nov. (strain CCIBt 3261). The generic name and specific epithets are proposed under the provisions of the International Code of Nomenclature for Algae, Fungi, and Plants.

Analysis of the toxicity and histopathology induced by the oral administration of Pseudanabaena galeata and Geitlerinema splendidum (cyanobacteria) extracts to mice.

Cyanobacteria are common members of the freshwater microbiota in lakes and drinking water reservoirs, and are responsible for several cases of human intoxications in Brazil. Pseudanabaena galeata and Geitlerinema splendidum are examples of the toxic species that are very frequently found in reservoirs in Sao Paulo, which is the most densely populated area in Brazil. In the search for toxic strains collected from water reservoirs and maintained in the Cyanobacterial Culture Collection (CCIBt) of the Institute of Botany of Brazil, the acetic acid extracts (AE) of P. galeata CCIBt 3082 and G. splendidum CCIBt 3223 were analyzed by planar chromatography, which indicated the absence of cyanotoxins. Animal tests were then carried out, and both extracts were found to induce toxic effects in mice when administered intraperitoneally. The present study aimed to investigate whether the oral ingestion of the above mentioned cyanobacteria extracts would also induce toxic effects in mice. Necropsy and histopathological studies were conducted using tissue samples from the animals, which were euthanized one week after the administration of the extracts. The AE of P. galeata did not cause death but did induce transient symptoms, including eyebrow ptosis, straub tail, and pain. The euthanized animals presented hemorrhage in the liver, whereas the histological analysis showed disorganization of the hepatic parenchyma, necrosis, hyperemia, and proximity of the centrilobular vein in the liver. In addition, alterations in the convoluted tubules of the kidneys were observed, and the lungs were unaffected. The AE of G. splendidum caused only one death, and induced transient symptoms, such as dyspnea, paralysis, and pain, in the other mice. The necropsy of the euthanized mice showed hemorrhage in the lungs and liver. The lungs presented hemorrhagic focuses, alveolar collapse, and granulomatous foci. The liver presented hemorrhagic and enlarged sinusoids, hyperemia, proximity of the centrilobular vein, and disorganization of the hepatic parenchyma. Some areas also exhibited an inflammatory infiltrate and calcified tissue inside blood vessels. Necrosis and rupture of the convoluted tubule cells were observed in the kidneys. Further analysis of the both extracts indicated the lack of hemolytic activity, and the presence of two unknown anti-AChE substances in the AE of G. splendidum. Thus, P. galeata and G. splendidum are producers of novel toxins that affect mammals when administered orally.

A toxic cyanobacterial bloom in an urban coastal lake, Rio Grande do Sul state, Southern Brazil.

Reports of cyanobacterial blooms developing worldwide have considerably increased, and, in most cases, the predominant toxins are microcystins. The present study reports a cyanobacterial bloom in Lake Violão, Torres, Rio Grande do Sul State, in January 2005. Samples collected on January 13, 2005, were submitted to taxonomical, toxicological, and chemical studies. The taxonomical analysis showed many different species of cyanobacteria, and that Microcystis protocystis and Sphaerocavum cf. brasiliense were dominant. Besides these, Microcystis panniformis, Anabaena oumiana, Cylindrospermopsis raciborskii, and Anabaenopsis elenkinii f. circularis were also present. The toxicity of the bloom was confirmed through intraperitoneal tests in mice, and chemical analyses of bloom extracts showed that the major substance was anabaenopeptin F, followed by anabaenopeptin B, microcystin-LR, and microcystin-RR.

Inflammatory effects of the toxic cyanobacterium Geitlerinema amphibium.

Toxic cyanobacteria in public water reservoirs may cause severe health issues for livestock and human beings. Geitlerinema amphibium, which is frequently found in São Paulo City's drinking water supplies, showed toxicity in the standard mouse bioassay, while displaying signs of intoxication and post-mortem findings different from those showed by animals intoxicated by known cyanotoxins. We report here the alterations caused by G. amphibium methanolic extract on mouse microcirculatory network, as seen by in vivo intravital microscopy, besides observations on leukocyte migration, cytokine quantitation, and results of toxicological essays. Our data showed that G. amphibium methanolic extract displayed time- and dose-dependent pro-inflammatory activity, and that at lower doses [125 and 250 mg/kg body weight (b.w.)] increased the leukocyte rolling, caused partial venular stasis, as well as induced an increase in leukocyte counts in the peripheral blood and peritoneal washings. At higher doses (500 and 1000 mg/kg b.w.), the extract caused ischemic injury leading to animal death. As confirmed by mass spectrometric studies and polymyxin B test, the G. amphibium methanolic extract did not contain lipopolysaccharides.

RELEASE OF CARBOHYDRATES AND PROTEINS BY A SUBTROPICAL STRAIN OF RAPHIDIOPSIS BROOKII (CYANOBACTERIA) ABLE TO PRODUCE SAXITOXIN AT THREE NITRATE CONCENTRATIONS(1).

Raphidiopsis brookii P. J. Hill (cyanobacteria) was isolated from a small subtropical eutrophic pond (Biguá Pond) located in the grounds of Rio Grande University in the extreme south of Brazil, following a toxic bloom of this species. Growth, saxitoxin production, and release of carbohydrates and protein were monitored at three sodium nitrate concentrations (500, 1,000, and 1,500 µM), from inoculation up to the stationary growth phase. Growth was monitored by determining the biovolume, chl content, and trichome count. Growth was better described in terms of biovolume and chl measurements, because trichome fragmentation was observed to increase at the stationary growth phase. Carbohydrates and proteins were released in small amounts during most of the experiment, with a significant increase during the stationary phase. Extracellular polysaccharides were essentially composed of glucose, galactose, N-acetyl-glucosamine, mannose, xylose, rhamnose, arabinose, and fucose. The relative proportions of these units showed no significant variation during growth. Small quantities of extracellular free carbohydrates were also detected, and only fucose was released in significant amounts at the lowest nitrate concentration (500 µM). R. brookii produced both saxitoxin and dc-saxitoxin, the former at four times the rate of the latter. This was the first study demonstrating saxitoxin production and the release of both carbohydrate and protein by R. brookii.

Lista de Cyanobacteria do Estado de São Paulo / Checklist of Cyanobacteria from São Paulo State, Brazil

Com base no levantamento bibliográfico realizado sobre as cianobactérias citadas para o Brasil e para o Estado de São Paulo em particular, além de consulta à lista de espécies da flora brasileira e aos bancos de dados de coleções paulistas, encontramos um total de 460 espécies citadas para o Brasil e 378 para o estado de São Paulo. Considerando que o grupo tem ao redor de 2800 espécies, estes números representam bem menos de 20% das espécies conhecidas. Assim, frente a diversidade de ambientes e habitats existentes nos biomas tropicais/subtropicais, o reduzido número de espécies já conhecidas indica que certamente essa biodiversidade está subestimada e deve ser muito maior do que identificamos até agora.

Based on the literature about the cyanobacteria from Brazil and particularly from São Paulo State, the list of Brazilian flora and the data bank of herbaria collections, a total of 460 species were referred to Brazil and 378 to São Paulo State. Taking into account that the group of cyanobacteria presents around 2800 species, these numbers represent much less than 20% of the known species. Considering the diversity of environments and habitats in the tropical/subtropical biomes compared to the reduced number of known species, this biodiversity is certainly underestimated and should be much larger.

A toxic cyanobacterial bloom in an urban coastal lake, Rio Grande do Sul state, Southern Brazil / Uma floração de cianobacterias em um lago costeiro urbano, Rio Grande do Sul, Brasil

Reports of cyanobacterial blooms developing worldwide have considerably increased, and, in most cases, the predominant toxins are microcystins. The present study reports a cyanobacterial bloom in Lake Violão, Torres, Rio Grande do Sul State, in January 2005. Samples collected on January 13, 2005, were submitted to taxonomical, toxicological, and chemical studies. The taxonomical analysis showed many different species of cyanobacteria, and that Microcystis protocystis and Sphaerocavum cf. brasiliense were dominant. Besides these, Microcystis panniformis, Anabaena oumiana,Cylindrospermopsis raciborskii, and Anabaenopsis elenkinii f. circularis were also present. The toxicity of the bloom was confirmed through intraperitoneal tests in mice, and chemical analyses of bloom extracts showed that the major substance was anabaenopeptin F, followed by anabaenopeptin B, microcystin-LR, and microcystin-RR.

O número de relatos de ocorrências de florações de cianobactérias em todo o mundo vem aumentando consideravelmente e na maioria desses episódios, as toxinas dominantes são as microcistinas. O presente estudo relata a ocorrência de floração na Lagoa do Violão, município de Torres, RS, em janeiro de 2005. As amostras coletadas em 13/01/2005 foram submetidas a estudos taxonômicos, toxicológicos e químicos. O exame microscópico do fitoplancton mostrou a dominância das espécies Microcystis protocystis e Sphaerocavum cf. brasiliense; foram observadas, também, Microcystis panniformis, Anabaena oumiana,Cylindrospermopsis raciborskii e Anabaenopsis elenkinii f. circularis. A toxicidade da floração foi confirmada através de ensaio intraperitonial em camundongos e a análise química de extratos obtidos da biomassa liofilizada mostrou que a substância majoritária era a anabaenopeptina F, seguida por anabaenopeptina B, microcistina-LR e microcistina-RR.

Inflammatory effects of the toxic cyanobacterium Geitlerinema amphibium

Toxic cyanobacteria in public water reservoirs may cause severe health issues for livestock and human beings. Geitlerinema amphibium, which is frequently found in São Paulo City’s drinking water supplies, showed toxicity in the standard mouse bioassay, while displaying signs of intoxication and post-mortem findings different from those showed by animals intoxicated by known cyanotoxins. We report here the alterations caused by G. amphibium methanolic extract on mouse microcirculatory network, as seen by in vivo intravital microscopy, besides observations on leukocyte migration, cytokine quantitation, and results of toxicological essays. Our data showed that G. amphibium methanolic extract displayed time- and dose-dependent pro-inflammatory activity, and that at lower doses [125 and 250 mg/kg body weight (b.w.)] increased the leukocyte rolling, caused partial venular stasis, as well as induced an increase in leukocyte counts in the peripheral blood and peritoneal washings. At higher doses (500 and 1000 mg/kg b.w.), the extract caused ischemic injury leading to animal death. As confirmed by mass spectrometric studies and polymyxin B test, the G. amphibium methanolic extract did not contain lipopolysaccharides.

Estudo qualitativo do fitoplâncton da represa de Serraria, Estado de Säo Paulo, Brasil / Qualitative study of phytoplankton from Serraria reservoir, state of Säo Paulo, Brazil

Durante o ano de 1979, foram realizadas coletas para o estudo qualitativo do fitoplâncton da Represa de Serraria, Estado de Säo Paulo, Brasil. Em cada coleta foram obtidas com garrafa Van Dorn, 6 amostras, nas profundidades correspondentes a 100, 50, 25, 10, 1 e 0% (zona afótica) de penetraçäo de luz, além de mais duas amostras, coletadas em toda d'água com redes de 20 e 53 micronm de abertura de malha. Foram identificados 108 táxons, sendo 45 de Chlorophyceae, 18 de Cyanophyceae, 18 de Zygnemaphyceae, 8 de Euglenophyceae, 6 de Diatomophyceae, 6 de Dinophyceae, 5 de Chrysophyceae, 1 de Oedogoniophyceae e 1 de Xanthophyceae. Uma nova forma de Staurastrum ellipticum W. West foi descrita neste trabalho

Algae and environment: a general approach

As espécies indicadoras de poluiçäo hídrica säo resistentes. A poluiçäo tem vários graus de intensidade, dependendo da diluiçäo do poluente no ambiente, fenômenos de antagonismo, sinergismo, grau de toxidade, etc, afetando a diversidade e a estabilidade dos organismos aquáticos. Para caracterizar zonas de poluiçäo säo usados grupos de organismos aquáticos do zoo e fitoplâncton. No Brasil, para as algas, há dificuldades por causa da falta de especialistas e recursos para a ciência taxonômica. Este trabalho faz levantamento histórico e recomenda a criaçäo de Centro Nacional de Taxonomia, para intercambiar informaçöes e facilitar o uso de algas como indicadores de poluiçäo

Toxic algae in freshwaters of Säo Paulo state

Sintetiza conhecimentos sobre floraçöes de algas e toxidade em corpos d'água no estado de Säo Paulo, e na literatura mundial. Discute a relaçäo entre eutrofizaçäo e ocorrência dessas floraçöes e enfatiza aspectos da soluçäo do problema