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Information on pollutant trophodynamics can be crucial for public health, as contaminated food consumption may lead to deleterious effects. This study was performed in Puruzinho Lake, a remote body of water in the Brazilian Amazon from which a riparian human population obtains an important part of its animal protein intake. Samples from 92 individuals, comprising 13 species and four trophic guilds (iliophagous, planktivorous, omnivorous, and piscivorous fish) were analysed for the determination of trace elements (Fe, Cr, Mn, Ni, Zn, Ca, Sr, Cd, Sn, Tl and Pb) and methylmercury concentrations. Samples from the same individuals had already been analysed for stable isotope (SI) measurements (δ13C and δ15N) in a previous investigation and the SI data have been statistically treated with those generated in this study for the evaluation of trophic dynamics of contaminants. Methylmercury was the only analyte that biomagnified, presenting TMF values of 4.65 and 4.55 for total and resident ichthyofauna, respectively. Trace elements presented either trophic dilution or independence from the trophic position, constituting a behaviour that was coherent with that found in the scientific literature. The similarity between Ni behaviour through the trophic web to that of essential elements contributes to the discussion on the essentiality of this metal to fish. Considering the Non-cancer Risk Assessment, the calculated Target Hazard Quotient (THQ) values were higher than 1.0 for all analysed individuals for methylmercury, as well as for only one individual for nickel. No other analyte rendered THQ values higher than 1.0.

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Holobionts are characterized by the relationship between host and their associated organisms such as the biofilm associated with macroalgae. Considering that light is essential to macroalgae survival, the aim of this study was to verify the effect of light on the heterotrophic activity in biofilms of the brown macroalgae Sargassum furcatum during its growth cycle. Measurements of heterotrophic activity were done under natural light levels at different times during a daily cycle and under an artificial extinction of natural light during the afternoon. We also measured Sargassum primary production under these light levels in the afternoon. Both measurements were done with and without photosynthesis inhibitor and antibiotics. Biofilm composition was mainly represented by bacteria but diatoms, cyanobacteria, and other organisms were also common. When a peak of diatom genera was recorded, the heterotrophic activity of the biofilm was higher. Heterotrophic activity was usually highest during the afternoon and the presence of a photosynthesis inhibitor caused an average reduction of 17% but there was no relationship with Sargassum primary production. These results indicate that autotrophic production in the biofilm was reduced by the inhibitor with consequences on bacterial activity. Heterotrophic activity was mainly bacterial and the antibiotics chloramphenicol and penicillin were more effective than streptomycin. We suggest primary producers in the biofilm are more important to increase bacterial activity than the macroalgae itself because of coherence of the peaks of heterotrophic and autotrophic activity in biofilm during the afternoon and the effects of autotrophic inhibitors on heterotrophic activity.

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Upwelling systems contain a high diversity of pelagic microorganisms and their composition and activity are defined by factors like temperature and nutrient concentration. Denaturing gradient gel electrophoresis (DGGE) technique was used to verify the spatial and temporal genetic variability of Bacteria and Archaea in two stations of the Arraial do Cabo coastal region, one under upwelling pressure and another under anthropogenic pressure. In addition, biotic and abiotic variables were measured in surface and deep waters from three other stations between these stations. Six samplings were done during a year and adequately represented the degrees of upwelling and anthropogenic pressures to the system. Principal Component Analysis (PCA) showed negative correlations between the concentrations of ammonia and phosphorous with prokaryotic secondary production and the total heterotrophic bacteria. PCA also showed negative correlation between temperature and the abundance of prokaryotic cells. Bacterial and archaeal compositions were changeable as were the oceanographic conditions, and upwelling had a regional pressure while anthropogenic pressure was punctual. We suggest that the measurement of prokaryotic secondary production was associated with both Bacteria and Archaea activities, and that substrate availability and temperature determine nutrients cycling.

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Arraial do Cabo is where upwelling occurs more intensively on the Brazilian coast. Although it is a protection area it suffers anthropogenic pressure such as harbor activities and sporadic sewage emissions. Short-time studies showed a high variability of bacterial production (BP) in this region but none of them evaluated BP during long periods in a large spatial scale including stations under different natural (upwelling and cold fronts) and anthropogenic pressures. During 2006, we sampled surface waters 10 times (5 in upwelling and 5 in subsidence periods) in 8 stations and we measured BP, temperature as well as the concentrations of inorganic nutrients, pigments and particulate organic matter (POM). BP was up to 400 times higher when sewage emissions were observed visually and it had a positive correlation with ammonia concentrations. Therefore, in 2007, we did two samples (each during upwelling and subsidence periods) during sewage emissions in five stations under different anthropogenic pressure and we also measured particles abundance by flow cytometry. The 12 samples in the most impacted area confirmed that BP was highest when ammonia was higher than 2 μM, also reporting the highest concentrations of chlorophyll a and suspended particles. However, considering all measured variables, upwelling was the main disturbing factor but the pressure of fronts should not be neglected since it had consequences in the auto-heterotrophic coupling, increasing the concentrations of non fluorescent particles and POM. Stations clustered in function of natural and anthropogenic pressures degrees and both determined the temporal-spatial variability.

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Arraial do Cabo is where upwelling occurs more intensively on the Brazilian coast. Although it is a protection area it suffers anthropogenic pressure such as harbor activities and sporadic sewage emissions. Short-time studies showed a high variability of bacterial production (BP) in this region but none of them evaluated BP during long periods in a large spatial scale including stations under different natural (upwelling and cold fronts) and anthropogenic pressures. During 2006, we sampled surface waters 10 times (5 in upwelling and 5 in subsidence periods) in 8 stations and we measured BP, temperature as well as the concentrations of inorganic nutrients, pigments and particulate organic matter (POM). BP was up to 400 times higher when sewage emissions were observed visually and it had a positive correlation with ammonia concentrations. Therefore, in 2007, we did two samples (each during upwelling and subsidence periods) during sewage emissions in five stations under different anthropogenic pressure and we also measured particles abundance by flow cytometry. The 12 samples in the most impacted area confirmed that BP was highest when ammonia was higher than 2 µM, also reporting the highest concentrations of chlorophyll a and suspended particles. However, considering all measured variables, upwelling was the main disturbing factor but the pressure of fronts should not be neglected since it had consequences in the auto-heterotrophic coupling, increasing the concentrations of non fluorescent particles and POM. Stations clustered in function of natural and anthropogenic pressures degrees and both determined the temporal-spatial variability.

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The ecological interaction between microorganisms and seaweeds depends on the production of secondary compounds that can influence microbial diversity in the water column and the composition of reef environments. We adapted the (3)H-leucine incorporation technique to measure bacterial activity in biofilms associated with the blades of the macroalgae Sargassum spp. We evaluated (1) if the epiphytic bacteria on the blades were more active in detritus or in the biofilm, (2) substrate saturation and linearity of (3)H-leucine incorporation, (3) the influence of specific metabolic inhibitors during (3)H-leucine incorporation under the presence or absence of natural and artificial light, and (4) the efficiency of radiolabeled protein extraction. Scanning electron microscopy showed heterogeneous distribution of bacteria, diatoms, and polymeric extracellular secretions. Active bacteria were present in both biofilm and detritus on the blades. The highest (3)H-leucine incorporation was obtained when incubating blades not colonized by macroepibionts. Incubations done under field conditions reported higher (3)H-leucine incorporation than in the laboratory. Light quality and sampling manipulation seemed to be the main factors behind this difference. The use of specific metabolic inhibitors confirmed that bacteria are the main group incorporating (3)H-leucine but their association with primary production suggested a symbiotic relationship between bacteria, diatoms, and the seaweed.

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Methylmercury (MeHg) increases mercury (Hg) toxicity and is biomagnified in the trophic chain contaminating riverine Amazon populations. Freshwater macrophyte roots are a main site of Hg methylation in different Brazilian environments. Paspalum repens periphyton was sampled in four floodplain lakes during the dry, rainy and wet seasons for measurement of total Hg (THg), MeHg, Hg methylation potentials, %C, %N, δ(13)C, δ(15)N and bacterial heterotrophic production as (3)H-leucine incorporation rate. THg concentration varied from 67 to 198 ng/g and the potential of Me(203)Hg formation was expressive (1-23%) showing that periphyton is an important matrix both in the accumulation of Hg and in MeHg production. The concentration of MeHg varied from 1 to 6 ng/g DW and was positively correlated with Me(203)Hg formation. Though methylmercury formation is mainly a bacterial process, no significant correlation was observed between the methylation potentials and bacterial production. The multiple regressions analyses suggested a negative correlation between THg and %C and %N and between methylation potential and δ(13)C. The discriminant analysis showed a significant difference in periphyton δ(15)N, δ(13)C and THg between seasons, where the rainy season presented higher δ(15)N and the wet period lighter δ(13)C, lower THg values and higher Me(203)Hg formation. This exploratory study indicates that the flooding cycle could influence the periphyton composition, mercury accumulation and methylmercury production.

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BACKGROUND: Upwelling systems are characterised by an intense primary biomass production in the surface (warmest) water after the outcrop of the bottom (coldest) water, which is rich in nutrients. Although it is known that the microbial assemblage plays an important role in the food chain of marine systems and that the upwelling systems that occur in southwest Brazil drive the complex dynamics of the food chain, little is known about the microbial composition present in this region. METHODOLOGY/PRINCIPAL FINDINGS: We carried out a molecular survey based on SSU rRNA gene from the three domains of the phylogenetic tree of life present in a tropical upwelling region (Arraial do Cabo, Rio de Janeiro, Brazil). The aim was to analyse the horizontal and vertical variations of the microbial composition in two geographically close areas influenced by anthropogenic activity (sewage disposal/port activity) and upwelling phenomena, respectively. A lower estimated diversity of microorganisms of the three domains of the phylogenetic tree of life was found in the water of the area influenced by anthropogenic activity compared to the area influenced by upwelling phenomena. We observed a heterogenic distribution of the relative abundance of taxonomic groups, especially in the Archaea and Eukarya domains. The bacterial community was dominated by Proteobacteria, Cyanobacteria and Bacteroidetes phyla, whereas the microeukaryotic community was dominated by Metazoa, Fungi, Alveolata and Stramenopile. The estimated archaeal diversity was the lowest of the three domains and was dominated by uncharacterised marine Crenarchaeota that were most closely related to Marine Group I. CONCLUSIONS/SIGNIFICANCE: The variety of conditions and the presence of different microbial assemblages indicated that the area of Arraial do Cabo can be used as a model for detailed studies that contemplate the correlation between pollution-indicating parameters and the depletion of microbial diversity in areas close to anthropogenic activity; functional roles and geochemical processes; phylogeny of the uncharacterised diversity; and seasonal variations of the microbial assemblages.

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The methylated form of mercury (Hg), methylmercury (MeHg), is one of the most toxic pollutants. Biotic and/or abiotic methylation, often associated to sulfate-reducing bacteria metabolism, occurs in aquatic environments and in many tropical areas, mostly in the periphyton associated to floating macrophyte roots. Data about mercury methylation by phytoplankton are scarce and the aim of this study was to verify the biotic influence in the methylation process in Microcystis aeruginosa and Sineccocystis sp. laboratory strains and in natural populations of phytoplankton from two different aquatic systems, the mesotrophic Ribeirão das Lajes reservoir and hypereutrophic oligohaline Jacarepaguá lagoon, Rio de Janeiro state, Brazil. Adapted radiochemical techniques were used to measure sulfate-reduction, mercury methylation and bacterial activity in phytoplankton samples. Methyl-(203)Hg formation from added inorganic (203)Hg and (3)H-Leucine uptake were measured by liquid scintillation as well as sulfate-reduction, estimated as H(2)(35)S produced from added Na(2)(35)SO(4). There was no significant difference in low methylation potentials (0.37%) among the two cyanobacterium species studied in laboratory conditions. At Ribeirão das Lajes reservoir, there was no significant difference in methylation, bacterial activity and sulfate-reduction of surface sediment between the sampling points. Methylation in sediments (3-4%) was higher than in phytoplankton (1.5%), the opposite being true for bacterial activity (sediment mean 6.6 against 150.3 nmol gdw(-1) h(-1) for phytoplankton samples). At Jacarepaguá lagoon, an expressive bacterial activity (477.1 x 10(3) nmol gdw(-1) h(-1) at a concentration of 1000 nM leucine) and sulfate-reduction ( approximately 21% H(2)(35)S trapped) associated to phytoplankton (mostly cyanobacteria M. aeruginosa) was observed, but mercury methylation was not detected.

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