Microbiological Quality Assessment by PCR and Its Antibiotic Susceptibility in Mangrove Crabs (Ucides cordatus) from Guanabara Bay, Rio de Janeiro, Brazil.

The bacteriological quality of crabs from three different mangroves (Itaóca, Suruí, and Piedade) from Rio de Janeiro state, Brazil, was investigated using conventional and molecular methods. The results revealed high counts for total coliforms in meat and hepatopancreas samples. PCR analyses identified 25 Escherichia coli colonies in the Itaóca, Piedade, and Suruí samples, detecting 13 enterotoxigenic colonies and 9 enteroaggregative colonies. Respectively, 12, 11, and 21 Vibrio parahaemolyticus strains were detected in the Itaóca, Piedade, and Suruí samples. Two V. cholerae strains were detected in the Piedade samples. The E. coli strains isolated in the present study showed resistance to gentamicin. E. coli strains from the Piedade samples showed 33% resistance to chloramphenicol and the strains also showed multiresistance to several antimicrobial agents with a MAR index ranging from 0.12 to 0.31. Vibrio strains from Piedade, Itaóca, and Suruí showed 86%, 78%, and 85% resistance, respectively, to ampicillin. The isolated Vibrio strains showed multiresistance to several antimicrobial agents, with a MAR index ranging from 0.12 to 0.25. The presence of these organisms in crab meat is an indication of microbial contamination, which may pose health risks to consumers when improperly cooked.

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.

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.

A novel antimicrobial lectin from Eugenia malaccensis that stimulates cutaneous healing in mice model.

OBJECTIVE: The present work reports the purification and partial characterization of an antibacterial lectin (EmaL) obtained from Eugenia malaccensis seeds as well as the evaluation of its effect in the daily topical treatment of repairing process of cutaneous wounds in mice. MATERIALS AND METHODS: The cutaneous wound was produced by the incision of the skin and use of lectin in the treatment of mice cutaneous wounds was evaluated. Surgical wounds were treated daily with a topical administration of EmaL and parameters such as edema, hyperemia, scab, granulation and scar tissues as well as contraction of wounds were analyzed. RESULTS: A novel lectin, with a molecular mass of 14 kDa, was isolated from E. malaccensis using affinity chromatography. The lectin (EmaL) agglutinated glutaraldehyde-treated rabbit and human erythrocytes; the lectin-induced rabbit erythrocyte agglutination was inhibited by glucose, casein, ovalbumin and fetuin. Also, Emal was very effective in the inhibition of bacterial growth, with the best inhibition results obtained for Staphylococcus aureus. Inflammatory signals such as edema and hyperemia were statistically less intense when EmaL was applied compared to the control. The histopathological analysis showed that the treated injured tissue presented reepithelialization (complete or partial) and areas of transition more evidenced than those of the control group, especially due to well organized pattern of collagen fibers presented in the granulation fibrous tissue. CONCLUSION: Presented results are a preliminary indication of the pharmacological interest in using EmaL as antimicrobial agent and in the repairing process of cutaneous wounds.

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.

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.

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.

Diversity of bacteria in the marine sponge Aplysina fulva in Brazilian coastal waters.

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.

Influence of experimental variables on decoloration of azo reactive dyes by hydrogen peroxide and UV radiation.

This paper was intended to study decoloration of azo C.I. Reactive Red 243 under UV radiation in the presence of hydrogen peroxide. The H2O2/UV oxidation process is a feasible and effective alternative in the treatment of effluents that contain such recalcitrant substances as reactive dyes. A statistical factorial design at 2n was used to evaluate how the variables pH (3 and 11), H2O2 concentration (5 and 20 mM), urea concentration (5 and 15 g l(-1)), and NaCl concentration (2.5 and 3.5 g l(-1)) affected the decoloration process efficiency. The results were expressed as percent colour removal from the solution by using absorbance measurements. The H2O2/UV process proved to be effective in degrading reactive dyes, with approximately 99% colour removal being achieved under specific conditions. Hydrogen peroxide concentration had a positive effect and influenced a great deal. Sodium chloride (NaCl) concentration presented a negative effect on the experiments in which it reduced the percent colour removal from the reactive dye solutions. Solution pH and urea concentration did not present any significant effect on the decoloration process.

Flow cytometry assessment of bacterioplankton in tropical marine environments.

Flow cytometry was used to characterize bacterioplankton from two tropical environments in Brazil: the eutrophic Guanabara Bay and the oligotrophic southwest Atlantic Ocean. Bacterial abundance was evaluated by flow cytometry, and cells were stained with SYTO 13, allowing demonstration of differences in nucleic acid content. Bacterial production was also evaluated by means of 3H-leucine incorporation. Bacterial numbers were different for both sites. In Atlantic Ocean samples, we found a maximum of 5.50 x 10(5) cells ml(-1), and low nucleic acid content organisms predominated. In Guanabara Bay, bacterial numbers were one order of magnitude higher than in the ocean, and they varied from outer bay (1.01 x 10(6) cells ml(-1)) to inner bay (6.90 x 10(6) cells ml(-1)). Bacterial activity in ocean samples varied from 4.6 to 126 ng C l(-1) h(-1), while in the bay, mean values ranged from 1.95 microg C l(-1) h(-1) (outer bay) to 7.35 microg C l(-1) h(-1) (inner bay). Values found for both parameters are characteristic of different trophic situations. These results illustrate the utility of cytometric analyses of bacterioplankton populations in characterizing their large spatial and temporal scales of distribution in aquatic ecosystems.

Unstable angina: individualized stratification and prognosis.

OBJECTIVE: To analyse the methodology of risk stratification and the prognosis of patients admitted with unstable angina. POPULATION AND METHODS: This retrospective study involved a population of 68 patients (43 males and 25 females with a mean age of 65.8 +/- 9.8 years) consecutively admitted for suspected unstable angina during the year of 1996. Thirty six patients (52.9%) had angina at rest, 13 patients (19.1%) had both exertional and rest angina, 9 patients (13.2%) crescendo angina, 6 patients (8.8%) new onset exertional angina (less than 1 month), and 4 patients (5.8%) post-infarction angina (less than 2 weeks). The risk stratification was individualized. The coronary angiography (35 patients) was only performed when the medical therapy failed in patients with recurrent angina, or with proved ischemia after an exercise test and/or thallium 201 stress scintigraphy. Thirteen patients (19.1%) did not undergo these tests (advanced age and or bad general condition, or refusal). The follow-up of patients with and without ST-T changes was compared, as well as those revascularized versus non-revascularized. It was possible to achieve a mean follow-up of 13.7 +/- 6.2 months (3 to 25 months). RESULTS: The exercise test and/or thallium-201 stress scintigraphy were positive for myocardial ischemia in 28 pts (41.1%) and negative in 7 patients (10.2%). The coronary angiography revealed three-vessel coronary artery disease in 18 patients (26.4%), one vessel disease in 11 patients (16.1%) and two-vessel disease in 5 patients (7.3%). One patient had normal coronary arteries. Medical therapy was the initial approach. Coronary surgery was urgently performed in 3 patients and coronary angioplasty in 5 patients for refractory unstable angina. In the whole group coronary artery surgery was undertaken in 14 patients (20.5%) and coronary angioplasty in 12 patients (17.6%). A mean follow-up of 13.7 +/- 6.2 months was obtained in the 68 patients. During this period 6 patients (8.8%) died due to cardiac causes and 16 patients (23.5%) were readmitted: 8 patients (11.7%) for unstable angina, 5 patients (7.3%) for congestive heart failure and 3 patients (4.4%) for myocardial infarction. Fifty two patients (76.4%) remained free of cardiac events. The patients with transitory ST-T changes had more cardiac events (unstable angina, myocardial infarction, mortality) than the patients without ECG changes (13/30 vs 2/30, p = 0.003). When the revascularized patients were compared to the non revascularized no significant differences were observed regarding myocardial infarction and mortality, however revascularized pts had a less significant incidence of rehospitalization for unstable angina (0/26 vs 8/42 p = 0.02). CONCLUSIONS: An individualized strategy can be effective in pts with unstable angina. In this study 76.4% of patients remained free of cardiac events during the follow-up, 23.6% had severe cardiac events and the cardiac mortality was 8.8%. The patients with transitory ST-T changes had more cardiac events and worse prognosis. No patients significant difference was observed in the revascularized versus non revascularized patients for myocardial infarction and mortality; however, the revascularized patients had less significant incidence of rehospitalization for unstable angina.

Ascomycetous yeast communities of marine invertebrates in a southeast Brazilian mangrove ecosystem.

The ascomycetous yeast communities associated with 3 bivalve mollusk, and 4 crab species were studied in the mangrove at Coroa Grande on Sepetiba Bay in Rio de Janeiro, Brazil. These were made up mostly of diverse but sparse and apparently allochtonous yeast populations. The striking exception was a prevalent population of the species Kluyveromyces aestuarii, which predominated the yeast communities of 2 detritus feeding crabs, Sesarma rectum and Uca spp., and the shipworm Neoteredo reynei. However, K. aestuarii was absent from the omnivorous crabs Aratus pisonii and Goniopsis cruentata, and the clam Anomalocardia brasiliana, and was rare in the clam Tagelus plebeius from mostly submerged more sandy sediments. Pichia membranaefaciens, Candida valida-like, Candida krusei, Candida sorbosa, Candida colliculosa-like, Candida famata-like, Kloeckera spp., Candida guilliermondii, Candida albicans, Candida silvae, Geotrichum spp., Rhodotorula spp., Cryptococcus spp., and the methylotrophic yeast Candida boidinii were frequently isolated. The 322 ascomycetous yeast cultures representing 252 isolates from crabs and mollusks were classified as 40 species that fit standard descriptions, and 44 putative new species. The ascomycetous yeast communities of the mangrove ecosystem include many new biotypes that require better taxonomic definition.

Yeasts and coliform bacteria of water accumulated in bromeliads of mangrove and sand dune ecosystems of southeast Brazil.

Yeasts and coliform bacteria were isolated from water that accumulated in the central cups and adjacent leaf axilae of two bromeliads, Neoregelia cruenta of a coastal sand dune and Quesnelia quesneliana of a mangrove ecosystem near the city of Rio de Janeiro, Brazil. The mean total coliform counts were above 10,000 per 100 mL for waters of both plants, but the mean fecal coliform counts were only 74 per 100 mL for Q. quesneliana and mostly undetected in water from N. cruenta. Of 90 fecal coliform isolates, 51 were typical of Escherichia coli in colony morphology and indol, methyl red, Volges-Proskauer, and citrate (IMViC) tests. Seven representatives of the typical E. coli cultures were identified as this species, but the identifications of nine other coliform bacteria were mostly dubious. The yeast community of N. cruenta was typical of plant surfaces with basidiomycetous yeasts anamorphs, and the black yeast Aureobasidium pullulans was prevalent. Quesnelia quesneliana had a substantial proportion of ascomycetous yeasts and their anamorphs, including a probable new biotype of Saccharomyces unisporus. Our results suggested that the microbial communities in bromeliad waters are typically autochtonous and not contaminants.