Fábio Vieira de Araújo

Diversity of Bacteria in the Marine Sponge Aplysina fulva in Brazilian Coastal Waters

ABSTRACT Microorganisms can account for up to 60% of the fresh weight of marine sponges. Marine sponges have been hypothesized to serve as accumulation spots of particular microbial communities, but it is unknown to what extent these communities are directed by the organism or the site or occur randomly. To address this question, we assessed the composition of specific bacterial communities associated with Aplysina fulva, one of the prevalent sponge species inhabiting Brazilian waters. Specimens of A. fulva and surrounding seawater were collected in triplicate in shallow water at two sites, Caboclo Island and Tartaruga beach, Búzios, Brazil. Total community DNA was extracted from the samples using “direct” and “indirect” approaches. 16S rRNA-based PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analyses of the total bacterial community and of specific bacterial groups—Pseudomonas and Actinobacteria—revealed that the structure of these assemblages in A. fulva differed drastically from that observed in seawater. The DNA extraction methodology and sampling site were determinative for the composition of actinobacterial communities in A. fulva. However, no such effects could be gleaned from total bacterial and Pseudomonas PCR-DGGE profiles. Bacterial 16S rRNA gene clone libraries constructed from directly and indirectly extracted DNA did not differ significantly with respect to diversity and composition. Altogether, the libraries encompassed 15 bacterial phyla and the candidate division TM7. Clone sequences affiliated with the Cyanobacteria, Chloroflexi, Gamma- and Alphaproteobacteria, Actinobacteria, Bacteroidetes, and Acidobacteria were, in this order, most abundant. The bacterial communities associated with the A. fulva specimens were distinct and differed from those described in studies of sponge-associated microbiota performed with other sponge species.

Microbial Diversity of a Brazilian Coastal Region Influenced by an Upwelling System and Anthropogenic Activity

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.

Bacterial communities reflect the spatial variation in pollutant levels in Brazilian mangrove sediment.

The majority of oil from oceanic oil spills converges on coastal ecosystems such as mangrove forests. A major challenge to mangrove bioremediation is defining the mangrove’s pollution levels and measuring its recuperation from pollution. Bioindicators can provide a welcome tool for defining such recovery. To determine if the microbial profiles reflected variation in the pollutants, samples from different locations within a single mangrove with a history of exposure to oil were chemically characterised, and the microbial populations were evaluated by a comprehensive range of conventional and molecular methods. Multivariate ordination of denaturing gradient gel electrophoresis (DGGE) microbial community fingerprints revealed a pronounced separation between the sediment and rhizosphere samples for all analysed bacterial communities (Bacteria, Betaproteobacteria, Alphaproteobacteria, Actinobacteria and Pseudomonas). A Mantel test revealed significant relationships between the sediment chemical fertility and oil-derived pollutants, most of the bacterial community fingerprints from sediment samples, and the counts by different cultivation strategies. The level of total petroleum hydrocarbons was significantly associated with the Bacteria and Betaproteobacteria fingerprints, whereas anthracene and the total level of polycyclic aromatic hydrocarbons were associated with the Actinobacteria. These results show that microbial communities from the studied mangrove reflect the spatial variation of the chemicals in the sediment, demonstrating the specific influences of oil-derived pollutants.

Kazachstania bromeliacearum sp. nov., a yeast species from water tanks of bromeliads

Cultures of a novel nutritionally specialized, fermentative yeast species were isolated from 34 water tanks of five bromeliad species, two mangrove sediment samples and one swamp water sample in Rio de Janeiro, Brazil. Sequence analysis of the D1/D2 domains of the large subunit of the rRNA gene showed that the novel species belongs to the genus Kazachstania. The novel species differs from Kazachstania martiniae by 11 substitutions and 2 gaps in the sequence of the domains D1/D2 of the LSU rRNA gene. The name Kazachstania bromeliacearum sp. nov. is proposed for the novel species. The type strain is IMUFRJ 51496T (=CBS 7996T=DBVPG 6864T=UFMG BR-174T).

Spatial–temporal analysis of marine debris on beaches of Niterói, RJ, Brazil: Itaipu and Itacoatiara

In many areas of the world, studies of marine debris are conducted with an emphasis on analyzing their composition, quantification and distribution on sandy beaches. However, in Brazil, studies are still restricted to some areas of the coast, and the quantities and the spatial and temporal patterns are unknown. To enhance the marine debris information in these areas, we selected the Itaipu and Itacoatiara beaches in Niterói, RJ, to collect, quantify and qualify the solid residues present in their sands. We collected 12 samples and recorded 118.39 kg of residues in Itaipu and 62.94 kg in Itacoatiara. At both beaches, the largest portion of debris was located on the upper part of the beach. Several debris items were related to food and drink consumption on the beaches, which indicated the contribution of beach users to pollution. Most of the debris was plastic. The greatest amount of debris was found at Itaipu in January and February and at Itacoatiara in January and March, months related to both the holiday season and abundant rainfall. The results demonstrated the necessity to implement an Environmental Education project for these areas to reduce its degradation.

Evaluation of microplastics in Jurujuba Cove, Niterói, RJ, Brazil, an area of mussels farming.

Once non-biodegradable, microplastics remain on the environment absorbing toxic hydrophobic compounds making them a risk to biodiversity when ingested or filtered by organisms and entering in the food chain. To evaluate the potential of the contamination by microplastics in mussels cultivated in Jurujuba Cove, Niterói, RJ, waters of three stations were collected during a rain and dry seasons using a plankton net and later filtered. Microplastics were quantified and characterized morphologically and chemically. The results showed a high concentration of microplastics in both seasons with diversity of colors, types and sizes. Synthetic polymers were present in all samples. The presence of microplastics was probably due to a high and constant load of effluent that this area receives and to the mussel farming activity that use many plastic materials. Areas with high concentrations of microplastics could not be used for mussel cultivation due to the risk of contamination to consumers.

KLUYVEROMYCES AESTUARII, A POTENTIAL ENVIRONMENTAL QUALITY INDICATOR YEAST FOR MANGROVES IN THE STATE OF RIO DE JANEIRO, BRAZIL

Kluyveromyces aestuarii was found in sediments from 7 of 8 mangroves in Rio de Janeiro; and absent only at one site with heavy plastic bag pollution. Its presence suggests influence in other habitats from a mangrove and its absence in a mangrove suggests some non- fecal pollution or other habitat alteration.

The influence of the intensity of use, rainfall and location in the amount of marine debris in four beaches in Niteroi, Brazil: Sossego, Camboinhas, Charitas and Flechas

The presence of marine debris in coastal and oceanic regions is a worldwide and growing problem and own to different factors. In order to check the influence of some of these factors in the amount of debris in these areas, we quantified and identified marine debris found on sand of four beaches in the city of Niterói, RJ during dry and rainy seasons; two in oceanic region and two in Guanabara Bay, and observed the intensity of use of them by people. Our results showed that intensity of use and intensity of rain had influence in the presence and amount of solid waste collected. Environmental education campaigns and improvements in basic sanitation are extremely necessary to prevent the pollution of aquatic environments and get improvements on waste management in the cities of Niterói, RJ.

Bacterial and Archaeal Communities Variability Associated with Upwelling and Anthropogenic Pressures in the Protection Area of Arraial do Cabo (Cabo Frio region - RJ)

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.

Tank bromeliad water: similar or distinct environments for research of bacterial bioactives?

The Atlantic Rainforest does not have a uniform physiognomy, its relief determines different environmental conditions that define the composition of its flora and fauna. Within this ecosystem, bromeliads that form tanks with their leaves hold water reservoirs throughout the year, maintaining complex food chains, based mainly on autotrophic and heterotrophic bacteria. Some works concluded that the water held by tank bromeliads concentrate the microbial diversity of their ecosystem. To investigate the bacterial diversity and the potential biotechnology of these ecosystems, tank bromeliads of the Neoregelia cruenta species from the Atlantic Rainforest in Brazil were used as models for this research. Bacteria isolated from these models were tested for production of bioactive compounds. DGGE of the water held by tank bromeliads was performed in different seasons, locations and sun exposure to verify whether these environmental factors affect bacterial communities. The DGGE bands profile showed no grouping of bacterial community by the environmental factors tested. Most of the isolates demonstrated promising activities in the tests performed. Collectively, these results suggest that tank bromeliads of the N. cruenta species provide important habitats for a diverse microbial community, suggesting that each tank forms a distinct micro-habitat. These tanks can be considered excellent sources for the search for new enzymes and/or new bioactive composites of microbial origin.