Cyanobacteria from coastal lagoons of Southern Brazil: coccoid organisms

Considering the great ecological importance of the cyanobacteria and the need for more detailed information about these organisms in Brazilian waters, this paper provides taxonomic information about the unicellular cyanobacteria flora in lagoon systems along the coastal plains of Rio Grande do Sul State. Sampling was performed in different freshwater bodies along the eastern (Casamento Lake area) and western (near the city of Tapes) banks of the Patos Lagoon (30º40' S-30º10' S and 50º30' W-51º30' W). The samples were collected once in the rainy season and once in the dry season (from May 2003 to December 2003) using a plankton net (25 µm mesh) in pelagic and littoral zones, and by squeezing the submerged parts of aquatic macrophytes. Thirty one species belonging to the families Synechoccocaceae (7 taxa), Merismopediaceae (12 taxa), Chamaesiphonaceae (1 taxon), Microcystaceae (4) and Chroococcaceae (7 taxa) were identified. Among these species, five are reported for the first time in Rio Grande do Sul State: Chamaesiphon amethystinus (Rostafinski) Lemmermann, Chroococcus minimus (Keissler) Lemmermann, Coelomoron pusillum (Van Goor) Komárek, Coelosphaerium kuetzingeanum Naegeli, and Cyanodictyon tubiforme Cronberg.

Modified nano- and microcellulose based adsorption materials in water treatment

In recent decades, industrial activity growth and increasing water usage worldwide have led to the release of various pollutants, such as toxic heavy metals and nutrients, into the aquatic environment. Modified nanocellulose and microcellulose-based adsorption materials have the potential to remove these contaminants from aqueous solutions. The present research consisted of the preparation of five different nano/microcellulose-based adsorbents, their characterization, the study of adsorption kinetics and isotherms, the determination of adsorption mechanisms, and an evaluation of adsorbents’ regeneration properties. The same well known reactions and modification methods that were used for modifying conventional cellulose also worked for microfibrillated cellulose (MFC). The use of succinic anhydride modified mercerized nanocellulose, and aminosilane and hydroxyapatite modified nanostructured MFC for the removal of heavy metals from aqueous solutions exhibited promising results. Aminosilane, epoxy and hydroxyapatite modified MFC could be used as a promising alternative for H2S removal from aqueous solutions. In addition, new knowledge about the adsorption properties of carbonated hydroxyapatite modified MFC as multifunctional adsorbent for the removal of both cations and anions ions from water was obtained. The maghemite nanoparticles (Fe3O4) modified MFC was found to be a highly promising adsorbent for the removal of As(V) from aqueous solutions due to its magnetic properties, high surface area, and high adsorption capacity . The maximum removal efficiencies of each adsorbent were studied in batch mode. The results of adsorption kinetics indicated very fast removal rates for all the studied pollutants. Modeling of adsorption isotherms and adsorption kinetics using various theoretical models provided information about the adsorbent’s surface properties and the adsorption mechanisms. This knowledge is important for instance, in designing water treatment units/plants. Furthermore, the correspondence between the theory behind the model and properties of the adsorbent as well as adsorption mechanisms were also discussed. On the whole, both the experimental results and theoretical considerations supported the potential applicability of the studied nano/microcellulose-based adsorbents in water treatment applications.

Corticolous cyanobacteria from tropical forest remnants in northwestern São Paulo State, Brazil

Cyanobacteria are common in aquatic environments but are also well-adapted to terrestrial habitats where they are represented by a diversified flora. The present study aimed to contribute to our taxonomic knowledge of terrestrial cyanobacteria by way of a floristic survey of the main components of corticolous communities found in seasonal semideciduous forest fragments. Samples of visible growths of Cyanobacteria, algae, and bryophytes found on tree bark were randomly collected and their taxonomies examined. Eighteen species of Cyanobacteria were found belonging to the genera Aphanothece, Chroococcus, Lyngbya, Phormidium, Porphyrosiphon, Hapalosiphon, Hassalia, Nostoc, Scytonema, and Stigonema. Many genera and species observed in the present work have been reported in previous surveys of the aerophytic flora in several regions of the world, although six species were described only on the basis of populations found in the forest fragments studied, which highlights the importance of taxonomic studies of cyanobacteria in these habitats.

The genera Chara and Nitella (Chlorophyta, Characeae) in the subtropical Itaipu Reservoir, Brazil

The family Characeae, represented by two genera in Brazil, Chara and Nitella, is considered to include the closest living relatives of land plants, and its members play important ecological role in aquatic ecosystems. The present taxonomic survey of Chara and Nitella was performed in tributaries that join to form the Brazilian shore of the Itaipu Reservoir on the Paraná River. Thirteen species were recorded, illustrated, and described: C. braunii var. brasiliensis R.Bicudo, C. guairensis R.Bicudo, N. acuminata A.Braun ex Wallman, N. furcata (Roxburgh ex Bruzileus) C.Agardh, and N. subglomerata A.Braun, already cited for the reservoir, and C. hydropitys Reichenbach, C. rusbyana Howe, N. axillaris A.Braun, N. glaziovii G.Zeller, N. gracilis (Smith) C.Agardh, N. hyalina (DC.) C.Agardh, N. inversa Imahori, and N. microcarpa A.Braun that represent new occurrences for the Itaipu Reservoir and Paraná State. Among the species encountered, C. guairensis, N. furcata, and N. glaziovii are widely distributed, while C. hydropitys and C. rusbyana have more restricted distributions.

Temporal variations in the primary productivity of Eleocharis acutangula (Cyperaceae) in a tropical wetland environment

Wetland vegetation typically includes aquatic macrophytes with high primary production capacities. The present study investigated how hydrological variations affect biomass allocation and primary productivity in the emergent macrophyte Eleocharis acutangula (Roxb.) Schult. Eleocharis acutangula ramets were collected from the Campelo Lagoon flood plain (21°39'S, 41°12'W and 21°37S, 41°11'W) between March/2005 and February/2006. This region experienced an unusually short rainy period between November/2005 and February/2006 that generated atypically high primary production levels (128gDWm-2month-1) and total biomass gains (447gDWm-2) in May and June/2005 respectively. Our data indicated that primary production and biomass allocation were strongly influenced by variations in wetland water levels and that macrophytes quickly invested in biomass accumulation when surface water levels rised.

An overview of lignin metabolism and its effect on biomass recalcitrance

Lignin, after cellulose, is the second most abundant biopolymer on Earth, accounting for 30% of the organic carbon in the biosphere. It is considered an important evolutionary adaptation of plants during their transition from the aquatic environment to land, since it bestowed the early tracheophytes with physical support to stand upright and enabled long-distance transport of water and solutes by waterproofing the vascular tissue. Although essential for plant growth and development, lignin is the major plant cell wall component responsible for biomass recalcitrance to industrial processing. The fact that lignin is a non-linear aromatic polymer built with chemically diverse and poorly reactive linkages and a variety of monomer units precludes the ability of any single enzyme to properly recognize and degrade it. Consequently, the use of lignocellulosic feedstock as a renewable and sustainable resource for the production of biofuels and bio-based materials will depend on the identification and characterization of the factors that determine plant biomass recalcitrance, especially the highly complex phenolic polymer lignin. Here, we summarize the current knowledge regarding lignin metabolism in plants, its effect on biomass recalcitrance and the emergent strategies to modify biomass recalcitrance through metabolic engineering of the lignin pathway. In addition, the potential use of sugarcane as a second-generation biofuel crop and the advances in lignin-related studies in sugarcane are discussed.

A new species of Lemmermanniella (Cyanobacteria) from the Atlantic Rainforest, Brazil

The Brazilian Atlantic Rainforest is a highly heterogeneous ecosystem comprising large numbers of tropical and subtropical habitats favorable to the development of cyanobacteria. Studies on cyanobacteria in this ecosystem are still rare, however, especially those involving unicellular and colonial types. The high biodiversity and endemism of this biome has been extremely impacted and fragmented, and less than 10% of its original vegetation cover remains today. We describe here a new species of a colonial cyanobacteria, Lemmermanniella terrestris, found on dry soils in a subtropical region of the Atlantic Rainforest in the municipality of Cananéia in southern São Paulo State, Brazil. This new taxon demonstrated all of the diacritical features of the genus Lemmermanniella but, unlike the other species of the genus, it was growing on the soil surface and not in an aquatic environment. A set of morphological features, including colonies composed of subcolonies, and cell dimensions, shapes and contents distinguish it from other species of the genus. Considering that species of Lemmermanniella are found in very distinct habitats (such as thermal and brackish waters) and that they maintain the same life cycle described for the genus in all of those environments, the morphological structures of the colonies can be used as reliable markers for identifying the genus, and its species differ primarily in relation to the habitats they occupy.

Cyanobacteria from coastal lagoons in southern Brazil: non-heterocytous filamentous organisms

This study describes and illustrates non-heterocytous filamentous cyanobacteria found in lagoon systems on the coastal plains of Rio Grande do Sul State. Collections were carried out in different freshwater bodies along the eastern (Casamento Lake area) and western (Tapes City area) margins of the Patos Lagoon (UTM 461948-6595095 and 542910-6645535) using a plankton net (25 µm mesh) in pelagic and littoral zones as well as by squeezing submerged parts of aquatic macrophytes, during both the rainy and dry seasons, from May to December/2003. Twenty two species belonging to the families Phormidiaceae (eight taxa), Pseudanabaenaceae (seven taxa), Oscillatoriaceae (six taxa), and Spirulinaceae (one taxon) were identified. Among these species, five are reported for the first time from Rio Grande do Sul State: Leptolyngbya cebennensis, Microcoleus subtorulosus, Oscillatoria cf. anguina, O. curviceps and Phormidium formosum.

Extended hydrogen photoproduction by nitrogen-fixing cyanobacteria

Cyanobacteria are the only prokaryotic organisms performing oxygenic photosynthesis. They comprise a diverse and versatile group of organisms in aquatic and terrestrial environments. Increasing genomic and proteomic data launches wide possibilities for their employment in various biotechnical applications. For example, cyanobacteria can use solar energy to produce H2. There are three different enzymes that are directly involved in cyanobacterial H2 metabolism: nitrogenase (nif) which produces hydrogen as a byproduct in nitrogen fixation; bidirectional hydrogenase (hox) which functions both in uptake and in production of H2; and uptake hydrogenase (hup) which recycles the H2 produced by nitrogenase back for the utilization of the cell. Cyanobacterial strains from University of Helsinki Cyanobacteria Collection (UHCC), isolated from the Baltic Sea and Finnish lakes were screened for efficient H2 producers. Screening about 400 strains revealed several promising candidates producing similar amounts of H2 (during light) as the ΔhupL mutant of Anabaena PCC 7120, which is specifically engineered to produce higher amounts of H2 by the interruption of uptake hydrogenase. The optimal environmental conditions for H2 photoproduction were significantly different between various cyanobacterial strains. All suitable strains revealed during screening were N2-fixing, filamentous and heterocystous. The top ten H2 producers were characterized for the presence and activity of the enzymes involved in H2 metabolism. They all possess the genes encoding the conventional nitrogenase (nifHDK1). However, the high H2 photoproduction rates of these strains were shown not to be directly associated with the maximum capacities of highly active nitrogenase or bidirectional hydrogenase. Most of the good producers possessed a highly active uptake hydrogenase, which has been considered as an obstacle for efficient H2 production. Among the newly revealed best H2 producing strains, Calothrix 336/3 was chosen for further, detailed characterization. Comparative analysis of the structure of the nif and hup operons encoding the nitrogenase and uptake hydrogenase enzymes respectively showed minor differences between Calothrix 336/3 and other N2-fixing model cyanobacteria. Calothrix 336/3 is a filamentous, N2-fixing cyanobacterium with ellipsoidal, terminal heterocysts. A common feature of Calothrix 336/3 is that the cells readily adhere to substrates. To make use of this feature, and to additionally improve H2 photoproduction capacity of the Calothrix 336/3 strain, an immobilization technique was applied. The effects of immobilization within thin alginate films were evaluated by examining the photoproduction of H2 of immobilized Calothrix 336/3 in comparison to model strains, the Anabaena PCC 7120 and its ΔhupL mutant. In order to achieve optimal H2 photoproduction, cells were kept under nitrogen starved conditions (Ar atmosphere) to ensure the selective function of nitrogenase in reducing protons to H2. For extended H2 photoproduction, cells require CO2 for maintenance of photosynthetic activity and recovery cycles to fix N2. Application of regular H2 production and recovery cycles, Ar or air atmospheres respectively, resulted in prolongation of H2 photoproduction in both Calothrix 336/3 and the ΔhupL mutant of Anabaena PCC 7120. However, recovery cycles, consisting of air supplemented with CO2, induced a strong C/N unbalance in the ΔhupL mutant leading to a decrease in photosynthetic activity, although total H2 yield was still higher compared to the wild-type strain. My findings provide information about the diversity of cyanobacterial H2 capacities and mechanisms and provide knowledge of the possibilities of further enhancing cyanobacterial H2 production.

Oxygen Photoreduction in Cyanobacteria

Cyanobacteria are well-known for their role in the global production of O2 via photosynthetic water oxidation. However, with the use of light energy, cyanobacteria can also reduce O2. In my thesis work, I have investigated the impact of O2 photoreduction on protection of the photosynthetic apparatus as well as the N2-fixing machinery. Photosynthetic light reactions produce intermediate radicals and reduced electron carriers, which can easily react with O2 to generate various reactive oxygen species. To avoid prolonged reduction of photosynthetic components, cyanobacteria use “electron valves” that dissipate excess electrons from the photosynthetic electron transfer chain in a harmless way. In Synechocystis sp. PCC 6803, flavodiiron proteins Flv1 and Flv3 comprise a powerful electron sink redirecting electrons from the acceptor side of Photosystem I to O2 and reducing it directly to water. In this work, I demonstrate that upon Ci-depletion Flv1/3 can dissipate up to 60% of the electrons delivered from Photosystem II. O2 photoreduction by Flv1/3 was shown to be vital for cyanobacteria in natural aquatic environments and deletion of Flv1/3 was lethal for both Synechocystis sp. PCC 6803 and Anabaena sp. PCC 7120 under fluctuating light conditions. The lethal phenotype observed in the absence of Flv1/3 results from oxidative damage to Photosystem I, which appeared to be a primary target of reactive oxygen species produced upon sudden increases in light intensity. Importantly, cyanobacteria also possess other O2 photoreduction pathways which can protect the photosynthetic apparatus. This study demonstrates that respiratory terminal oxidases are also capable of initiating O2 photoreduction in mutant cells lacking the Flv1/3 proteins and grown under fluctuating light. Photoreduction of O2 by Rubisco was also shown in Ci-depleted cells of the mutants lacking Flv1/3, and thus provided the first evidence for active photorespiratory gas-exchange in cyanobacteria. Nevertheless, and despite the existence of other O2 photoreduction pathways, the Flv1/3 route appears to be the most robust and rapid system of photoprotection. Several groups of cyanobacteria are capable of N2 fixation. Filamentous heterocystous N2- fixing species, such as Anabaena sp. PCC 7120, are able to differentiate specialised cells called heterocysts for this purpose. In contrast to vegetative cells which perform oxygenic photosynthesis, heterocysts maintain a microoxic environment for the proper function of the nitrogenase enzyme, which is extremely sensitive to O2. The genome of Anabaena sp. PCC 7120 harbors two copies of genes encoding Flv1 and Flv3 proteins, designated as “A” and “B” forms. In this thesis work, I demonstrate that Flv1A and Flv3A are expressed only in the vegetative cells of filaments, whilst Flv1B and Flv3B are localized exclusively in heterocysts. I further revealed that the Flv3B protein is most responsible for the photoreduction of O2 in heterocysts, and that this reaction plays an important role in protection of the N2-fixing machinery and thus, the provision of filaments with fixed nitrogen. The function of the Flv1B protein remains to be elucidated; however the involvement of this protein in electron transfer reactions is feasible. Evidence provided in this thesis indicates the presence of a great diversity of O2 photoreduction reactions in cyanobacterial cells. These reactions appear to be crucial for the photoprotection of both photosynthesis and N2 fixation processes in an oxygenic environment.

Monoamines in the pedal plexus of the land snail Megalobulimus oblongus (Gastropoda, Pulmonata)

In molluscs, the number of peripheral neurons far exceeds those found in the central nervous system. Although previous studies on the morphology of the peripheral nervous system exist, details of its organization remain unknown. Moreover, the foot of the terrestrial species has been studied less than that of the aquatic species. As this knowledge is essential for our experimental model, the pulmonate gastropod Megalobulimus oblongus, the aim of the present study was to investigate monoamines in the pedal plexus of this snail using two procedures: glyoxylic acid histofluorescence to identify monoaminergic structures, and the unlabeled antibody peroxidase anti-peroxidase method using antiserum to detect the serotonergic component of the plexus. Adult land snails weighing 48-80 g, obtained from the counties of Barra do Ribeiro and Charqueadas (RS, Brazil), were utilized. Monoaminergic fibers were detected throughout the pedal musculature. Blue fluorescence (catecholamines, probably dopamine) was observed in nerve branches, pedal and subepithelial plexuses, and in the pedal muscle cells. Yellow fluorescence (serotonin) was only observed in thick nerves and in muscle cells. However, when immunohistochemical methods were used, serotonergic fibers were detected in the pedal nerve branches, the pedal and subepithelial plexuses, the basal and lateral zones of the ventral integument epithelial cells, in the pedal ganglion neurons and beneath the ventral epithelium. These findings suggest catecholaminergic and serotonergic involvement in locomotion and modulation of both the pedal ganglion interneurons and sensory information. Knowledge of monoaminergic distribution in this snail´s foot is important for understanding the pharmacological control of reflexive responses and locomotive behavior.

Cytotoxicity and genotoxicity of low doses of mercury chloride and methylmercury chloride on human lymphocytes in vitro

Mercury is a xenobiotic metal that is a highly deleterious environmental pollutant. The biotransformation of mercury chloride (HgCl2) into methylmercury chloride (CH3HgCl) in aquatic environments is well-known and humans are exposed by consumption of contaminated fish, shellfish and algae. The objective of the present study was to determine the changes induced in vitro by two mercury compounds (HgCl2 and CH3HgCl) in cultured human lymphocytes. Short-term human leukocyte cultures from 10 healthy donors (5 females and 5 males) were set-up by adding drops of whole blood in complete medium. Cultures were separately and simultaneously treated with low doses (0.1 to 1000 µg/l) of HgCl2 and CH3HgCl and incubated at 37ºC for 48 h. Genotoxicity was assessed by chromosome aberrations and polyploid cells. Mitotic index was used as a measure of cytotoxicity. A significant increase (P < 0.05) in the relative frequency of chromosome aberrations was observed for all concentrations of CH3HgCl when compared to control, whether alone or in an evident sinergistic combination with HgCl2. The frequency of polyploid cells was also significantly increased (P < 0.05) when compared to control after exposure to all concentrations of CH3HgCl alone or in combination with HgCl2. CH3HgCl significantly decreased (P < 0.05) the mitotic index at 100 and 1000 µg/l alone, and at 1, 10, 100, and 1000 µg/l when combined with HgCl2, showing a synergistic cytotoxic effect. Our data showed that low concentrations of CH3HgCl might be cytotoxic/genotoxic. Such effects may indicate early cellular changes with possible biological consequences and should be considered in the preliminary evaluation of the risks of populations exposed in vivo to low doses of mercury.

Environmental Challenges in the Joint Border Area of Norway, Finland and Russia

This report examines the human impact on the subarctic environment of the joint border area of Norway, Finland and Russia. The aim is to present the current state and recent changes that have taken place in the region. The main threat to the environment is the Pechenganikel mining and metallurgical industrial combine in the towns of Nikel and Zapolyarny in the Kola Peninsula. Emissions from this complex include high levels of heavy metals, persistent organic pollutants and sulfur dioxide. Pollution, along with climate change, water level regulation and other anthropogenic effects, has affected the aquatic ecosystems in the joint border area. The main heavy metals in the area are copper and nickel, the highest concentrations of which are measured near the combine. Direct discharge of sewage into the river continues and airborne heavy metal particles are also deposited to areas farther away. Climate changeinduced increase in temperature and precipitation in the Kola Peninsula is evident. Water level regulation with seven hydropower plants in the Pasvik River have changed it into a series of lakes and lake-like reservoirs. This report discusses modelling, which was enabled to estimate the effect of climate change on Lake Inarijärvi and the Pasvik River hydrology, water level fluctuation and ecology and to follow the sulfur dioxide emissions emitted from the Pechenganikel. Effects of pollution on the nature and concentrations of the main pollutants were studied and climate change in the border area and its effects on the ecology were estimated. Also the effects of water level regulation on the ecological status of the aquatic ecosystems were addressed.

Spatial and temporal variation in fish populations and assemblages in coastal waters of the northern Baltic Proper

The distribution and traits of fish are of interest both ecologically and socio-economically. In this thesis, phenotypic and structural variation in fish populations and assemblages was studied on multiple spatial and temporal scales in shallow coastal areas in the archipelago of the northern Baltic Proper. In Lumparn basin in Åland Islands, the fish assemblage displayed significant seasonal variation in depth zone distribution. The results indicate that investigating both spatial and temporal variation in small scale is crucial for understanding patterns in fish distribution and community structure in large scale. The local population of Eurasian perch Perca fluviatilis L displayed habitat-specific morphological and dietary variation. Perch in the pelagic zone were on average deeper in their body shape than the littoral ones and fed on fish and benthic invertebrates. The results differ from previous studies conducted in freshwater habitats, where the pelagic perch typically are streamlined in body shape and zooplanktivorous. Stable isotopes of carbon and nitrogen differed between perch with different stomach contents, suggesting differentiation of individual diet preferences. In the study areas Lumparn and Ivarskärsfjärden in Åland Islands and Galtfjärden in Swedish east coast, the development in fish assemblages during the 2000’s indicated a general shift towards higher abundances of small-bodied lower-order consumers, especially cyprinids. For European pikeperch Sander lucioperca L., recent declines in adult fish abundances and high mortalities (Z = 1.06–1.16) were observed, which suggests unsustainably high fishing pressure on pikeperch. Based on the results it can be hypothesized that fishing has reduced the abundances of large predatory fish, which together with bottom-up forcing by eutrophication has allowed the lower-order consumer species to increase in abundances. This thesis contributes to the scientific understanding of aquatic ecosystems with new descriptions on morphological and dietary adaptations in perch in brackish water, and on the seasonal variation in small-scale spatial fish distribution. The results also demonstrate anthropogenic effects on coastal fish communities and underline the urgency of further reducing nutrient inputs and regulating fisheries in the Baltic Sea region.

Control of respiration in fish, amphibians and reptiles

Fish and amphibians utilise a suction/force pump to ventilate gills or lungs, with the respiratory muscles innervated by cranial nerves, while reptiles have a thoracic, aspiratory pump innervated by spinal nerves. However, fish can recruit a hypobranchial pump for active jaw occlusion during hypoxia, using feeding muscles innervated by anterior spinal nerves. This same pump is used to ventilate the air-breathing organ in air-breathing fishes. Some reptiles retain a buccal force pump for use during hypoxia or exercise. All vertebrates have respiratory rhythm generators (RRG) located in the brainstem. In cyclostomes and possibly jawed fishes, this may comprise elements of the trigeminal nucleus, though in the latter group RRG neurons have been located in the reticular formation. In air-breathing fishes and amphibians, there may be separate RRG for gill and lung ventilation. There is some evidence for multiple RRG in reptiles. Both amphibians and reptiles show episodic breathing patterns that may be centrally generated, though they do respond to changes in oxygen supply. Fish and larval amphibians have chemoreceptors sensitive to oxygen partial pressure located on the gills. Hypoxia induces increased ventilation and a reflex bradycardia and may trigger aquatic surface respiration or air-breathing, though these latter activities also respond to behavioural cues. Adult amphibians and reptiles have peripheral chemoreceptors located on the carotid arteries and central chemoreceptors sensitive to blood carbon dioxide levels. Lung perfusion may be regulated by cardiac shunting and lung ventilation stimulates lung stretch receptors.