Efeitos do exercício intenso e do condicionamento aeróbio associado a suplementação com cromo ou carnitina sobre a microbiota fecal de potras

Pós-graduação em Microbiologia Agropecuária - FCAV

Detecção molecular de Ehrlichia canis em cães e em órgãos de seus respectivos carrapatos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Identificação taxonômica e análise conexionista de bactérias heterotróficas e fatores abióticos de ambientes lóticos sob influência de efluentes do refino no petróleo

This study used a multi-analytical approach based on traditional microbiological methods for cultivation and isolation of heterotrophic bacteria in the laboratory associated with the molecular identification of the isolates and physicochemical analysis of environmental samples. The model chosen for data integration was supported by knowledge from computational neuroscience, and composed by three modules: (i) microbiological parameters, contemplating taxonomic data obtained from the partial sequencing of the 16S rRNA gene from 80 colonies of heterotrophic bacteria isolated by plating method in PCA media. For bacterial colonies isolation were used water samples from Atibaia and Jaguarí rivers collected at the site of water captation for use in effluent treatment, upstream from the entrance of treated effluent from the Paulínia refinery (REPLAN/Petrobras) located in the Paulínia-SP municipality, from the output of the biological treatment plant with stabilization pond and from the raw refinery wastewater; (ii) chemical parameters, ending measures of dissolved oxygen (DO), chemical oxygen demand (COD), biochemical oxygen demand (BOD), chloride, acidity CaCO3, alkalinity, ammonia, nitrite, nitrate, dissolved ions, sulfides, oils and greases; and (iii) physical parameters, comprising the pH determination, conductivity, temperature, transparency, settleable solids, suspended and soluble solids, volatile material, remaining fixing material (RFM), apparent color and turbidity. The results revealed interesting theoretical relationships involving two families of bacteria (Carnobacteriaceae and Aeromonadaceae). Carnobacteriaceae revealed positive theoretical relationships with COD, BOD, nitrate, chloride, temperature, conductivity and apparent color and negative theoretical relationships with the OD. Positive theoretical relationships were shown between Aeromonadaceae and OD and nitrate, while this bacterial family showed negative theoretical...

Detecção molecular de Ehrlichia canis em cães e em órgãos de seus respectivos carrapatos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Identificação taxonômica e análise conexionista de bactérias heterotróficas e fatores abióticos de ambientes lóticos sob influência de efluentes do refino no petróleo

This study used a multi-analytical approach based on traditional microbiological methods for cultivation and isolation of heterotrophic bacteria in the laboratory associated with the molecular identification of the isolates and physicochemical analysis of environmental samples. The model chosen for data integration was supported by knowledge from computational neuroscience, and composed by three modules: (i) microbiological parameters, contemplating taxonomic data obtained from the partial sequencing of the 16S rRNA gene from 80 colonies of heterotrophic bacteria isolated by plating method in PCA media. For bacterial colonies isolation were used water samples from Atibaia and Jaguarí rivers collected at the site of water captation for use in effluent treatment, upstream from the entrance of treated effluent from the Paulínia refinery (REPLAN/Petrobras) located in the Paulínia-SP municipality, from the output of the biological treatment plant with stabilization pond and from the raw refinery wastewater; (ii) chemical parameters, ending measures of dissolved oxygen (DO), chemical oxygen demand (COD), biochemical oxygen demand (BOD), chloride, acidity CaCO3, alkalinity, ammonia, nitrite, nitrate, dissolved ions, sulfides, oils and greases; and (iii) physical parameters, comprising the pH determination, conductivity, temperature, transparency, settleable solids, suspended and soluble solids, volatile material, remaining fixing material (RFM), apparent color and turbidity. The results revealed interesting theoretical relationships involving two families of bacteria (Carnobacteriaceae and Aeromonadaceae). Carnobacteriaceae revealed positive theoretical relationships with COD, BOD, nitrate, chloride, temperature, conductivity and apparent color and negative theoretical relationships with the OD. Positive theoretical relationships were shown between Aeromonadaceae and OD and nitrate, while this bacterial family showed negative theoretical...

Bacterial diversity in rhizosphere soil from Antarctic vascular plants of Admiralty Bay, maritime Antarctica

The Antarctic is a pristine environment that contributes to the maintenance of the global climate equilibrium. The harsh conditions of this habitat are fundamental to selecting those organisms able to survive in such an extreme habitat and able to support the relatively simple ecosystems. The DNA of the microbial community associated with the rhizospheres of Deschampsia antarctica Desv (Poaceae) and Colobanthus quitensis (Kunth) BartI (Caryophyllaceae), the only two native vascular plants that are found in Antarctic ecosystems, was evaluated using a 16S rRNA multiplex 454 pyrosequencing approach. This analysis revealed similar patterns of bacterial diversity between the two plant species from different locations, arguing against the hypothesis that there would be differences between the rhizosphere communities of different plants. Furthermore, the phylum distribution presented a peculiar pattern, with a bacterial community structure different from those reported of many other soils. Firmicutes was the most abundant phylum in almost all the analyzed samples, and there were high levels of anaerobic representatives. Also, some phyla that are dominant in most temperate and tropical soils, such as Acidobacteria, were rarely found in the analyzed samples. Analyzing all the sample libraries together, the predominant genera found were Bifidobacterium (phylum Actinobacteria), Arcobacter (phylum Proteobacteria) and Faecalibacterium (phylum Firmicutes). To the best of our knowledge, this is the first major bacterial sequencing effort of this kind of soil, and it revealed more than expected diversity within these rhizospheres of both maritime Antarctica vascular plants in Admiralty Bay, King George Island, which is part of the South Shetlands archipelago. The ISME Journal (2010) 4, 989-1001; doi:10.1038/ismej.2010.35; published online 1 April 2010

Biogeography of two cold-adapted genera: Psychrobacter and Exiguobacterium

The genera Exiguobacterium and Psychrobacter have been frequently detected in and isolated from polar permafrost and ice. These two genera have members that can grow at temperatures as low as -5 and -10 degrees C, respectively. We used quantitative PCR (Q-PCR) to quantify members of these genera in 54 soil or sediment samples from polar, temperate and tropical environments to determine to what extent they are selected by cold environments. These results were further analyzed by multiple linear regression to identify the most relevant environmental factors corresponding to their distribution. Exiguobacterium was detected in all three climatic zones at similar densities, but was patchier in the temperate and tropical samples. Psychrobacter was present in almost all polar samples, was at highest densities in Antarctica sediment samples, but was in very low densities and infrequently detected in temperate and tropical soils. Clone libraries, specific for the 16S rRNA gene for each genus, were constructed from a sample from each climatic region. The clone libraries were analyzed for alpha and beta diversities, as well as for variation in population structure by using analysis of molecular variance. Results confirm that both genera were found in all three climatic zones; however, Psychrobacter populations seemed to be much more diverse than Exiguobacterium in all three climatic zones. Furthermore, Psychrobacter populations from Antarctica are different from those in Michigan and Puerto Rico, which are similar to each other. The ISME Journal (2009) 3, 658-665; doi: 10.1038/ismej.2009.25; published online 26 March 2009

Diversity of hydrocarbon-degrading Klebsiella strains isolated from hydrocarbon-contaminated estuaries

To investigate the diversity and the catabolic capacity of oil-degrading Klebsiella strains isolated from hydrocarbon-contaminated sediments in Santos-Sao Vicente estuary systems in Brazil. Klebsiella strains obtained from the estuary were characterized using 16S rRNA gene sequencing and BOX-PCR patterns, testing their catabolic capacity to degrade toluene, xylene, naphthalene and nonane, and identifying the catabolic genes present in the oil-degrading strains. Results show that Klebsiella strains were widespread in the estuary. Twenty-one isolates from the Klebsiella genus were obtained; 14 had unique BOX patterns and were further investigated. Among four distinct catabolic genes tested (todC1, ndoB, xylE and alkB1), only the todC1 gene could be amplified in two Klebsiella strains. The biodegradation assay showed that most of the strains had the ability to degrade all of the tested hydrocarbons; however, the strains displayed different efficiencies. The oil-degrading Klebsiella isolates obtained from the estuary were closely related to Klebsiella pneumoniae and Klebsiella ornithinolytica. The isolates demonstrated a substantial degree of catabolic plasticity for hydrocarbon degradation. The results of this study show that several strains from the Klebsiella genus are able to degrade diverse hydrocarbon compounds. These findings indicate that Klebsiella spp. can be an important part of the oil-degrading microbial community in estuarine areas exposed to sewage.

Gastrotricha: A Marine Sister for a Freshwater Puzzle

Background: Within an evolutionary framework of Gastrotricha Marinellina flagellata and Redudasys fornerise bear special interest, as they are the only Macrodasyida that inhabit freshwater ecosystems. Notwithstanding, these rare animals are poorly known; found only once (Austria and Brazil), they are currently systematised as incertae sedis. Here we report on the rediscovery of Redudasys fornerise, provide an account on morphological novelties and present a hypothesis on its phylogenetic relationship based on molecular data. Methodology/Principal Findings: Specimens were surveyed using DIC microscopy and SEM, and used to obtain the 18 S rRNA gene sequence; molecular data was analyzed cladistically in conjunction with data from 42 additional species belonging to the near complete Macrodasyida taxonomic spectrum. Morphological analysis, while providing new information on taxonomically relevant traits (adhesive tubes, protonephridia and sensorial bristles), failed to detect elements of the male system, thus stressing the parthenogenetic nature of the Brazilian species. Phylogenetic analysis, carried out with ML, MP and Bayesian approaches, yielded topologies with strong nodal support and highly congruent with each other. Among the supported groups is the previously undocumented clade showing the alliance between Redudasys fornerise and Dactylopodola agadasys; other strongly sustained clades include the densely sampled families Thaumastodermatidae and Turbanellidae and most genera. Conclusions/Significance: A reconsideration of the morphological traits of Dactylopodola agadasys in light of the new information on Redudasys fornerise makes the alliance between these two taxa very likely. As a result, we create Anandrodasys gen. nov. to contain members of the previously described D. agadasys and erect Redudasyidae fam. nov. to reflect this novel relationship between Anandrodasys and Redudasys. From an ecological perspective, the derived position of Redudasys, which is deeply nested within the Macrodasyida clade, unequivocally demonstrates that invasion of freshwater by gastrotrichs has taken place at least twice, in contrast with the single event hypothesis recently put forward.

Chromatin Structure of Ribosomal RNA Genes in Dipterans and Its Relationship to the Location of Nucleolar Organizers

Nucleoli, nuclear organelles in which ribosomal RNA is synthesized and processed, emerge from nucleolar organizers (NORs) located in distinct chromosomal regions. In polytene nuclei of dipterans, nucleoli of some species can be observed under light microscopy exhibiting distinctive morphology: Drosophila and chironomid species display well-formed nucleoli in contrast to the fragmented and dispersed nucleoli seen in sciarid flies. The available data show no apparent relationship between nucleolar morphology and location of NORs in Diptera. The regulation of rRNA transcription involves controlling both the transcription rate per gene as well as the proportion of rRNA genes adopting a proper chromatin structure for transcription, since active and inactive rRNA gene copies coexist in NORs. Transcription units organized in nucleosomes and those lacking canonical nucleosomes can be analyzed by the method termed psoralen gel retarding assay (PGRA), allowing inferences on the ratio of active to inactive rRNA gene copies. In this work, possible connections between chromosomal location of NORs and proportion of active rRNA genes were studied in Drosophila melanogaster, and in chironomid and sciarid species. The data suggested a link between location of NORs and proportion of active rRNA genes since the copy number showing nucleosomal organization predominates when NORs are located in the pericentric heterochromatin. The results presented in this work are in agreement with previous data on the chromatin structure of rRNA genes from distantly related eukaryotes, as assessed by the PGRA.

Ultraviolet-radiation-resistant isolates revealed cellulose-degrading species of Cellulosimicrobium cellulans (UVP1) and Bacillus pumilus (UVP4)

Among extremophiles, microorganisms resistant to ultraviolet radiation (UVR) have been known to produce a variety of metabolites (i.e., extremolytes). We hypothesized that natural microbial flora on elevated land (hills) would reveal a variety of UVR-resistant extremophiles and polyextremophiles with modulated proteins and enzymes that had biotechnological implications. Microorganisms Cellulosimicrobium cellulans UVP1 and Bacillus pumilus UVP4 were isolated and identified using 16S rRNA sequencing, and showed extreme UV resistance (1.03 x 106 and 1.71 x 105 similar to J/m2, respectively) from elevated land soil samples along with unique patterns of protein expression under UVR and non-UVR. A broad range of cellulolytic activity on carboxymethyl cellulose agar plates in C. cellulans UVP1 and B. pumilus UVP4 was revealed at varying pH, temperature, and inorganic salt concentration. Further, the microbial strain B. pumilus UVP4 showed the basic characteristics of a novel group: polyextremophiles with significance in bioenergy.

Eucalyptus growth promotion by endophytic Bacillus spp

Clonal eucalyptus plantings have increased in recent years; however, some clones with high production characteristics have vegetative propagation problems because of weak root and aerial development. Endophytic microorganisms live inside healthy plants without causing any damage to their hosts and can be beneficial, acting as plant growth promoters. We isolated endophytic bacteria from eucalyptus plants and evaluated their potential in plant growth promotion of clonal plantlets of Eucalyptus urophylla x E. grandis, known as the hybrid, E. urograndis. Eighteen isolates of E. urograndis, clone 4622, were tested for plant growth promotion using the same clone. These isolates were also evaluated for indole acetic acid production and their potential for nitrogen fixation and phosphate solubilization. The isolates were identified by partial sequencing of 16S rRNA. Bacillus subtilis was the most prevalent species. Several Bacillus species, including B. licheniformis and B. subtilis, were found for the first time as endophytes of eucalyptus. Bacillus sp strain EUCB 10 significantly increased the growth of the root and aerial parts of eucalyptus plantlets under greenhouse conditions, during the summer and winter seasons.

Structural and Functional Insights from the Metagenome of an Acidic Hot Spring Microbial Planktonic Community in the Colombian Andes

A taxonomic and annotated functional description of microbial life was deduced from 53 Mb of metagenomic sequence retrieved from a planktonic fraction of the Neotropical high Andean (3,973 meters above sea level) acidic hot spring El Coquito (EC). A classification of unassembled metagenomic reads using different databases showed a high proportion of Gammaproteobacteria and Alphaproteobacteria (in total read affiliation), and through taxonomic affiliation of 16S rRNA gene fragments we observed the presence of Proteobacteria, micro-algae chloroplast and Firmicutes. Reads mapped against the genomes Acidiphilium cryptum JF-5, Legionella pneumophila str. Corby and Acidithiobacillus caldus revealed the presence of transposase-like sequences, potentially involved in horizontal gene transfer. Functional annotation and hierarchical comparison with different datasets obtained by pyrosequencing in different ecosystems showed that the microbial community also contained extensive DNA repair systems, possibly to cope with ultraviolet radiation at such high altitudes. Analysis of genes involved in the nitrogen cycle indicated the presence of dissimilatory nitrate reduction to N2 (narGHI, nirS, norBCDQ and nosZ), associated with Proteobacteria-like sequences. Genes involved in the sulfur cycle (cysDN, cysNC and aprA) indicated adenylsulfate and sulfite production that were affiliated to several bacterial species. In summary, metagenomic sequence data provided insight regarding the structure and possible functions of this hot spring microbial community, describing some groups potentially involved in the nitrogen and sulfur cycling in this environment. Citation: Jimenez DJ, Andreote FD, Chaves D, Montana JS, Osorio-Forero C, et al. (2012) Structural and Functional Insights from the Metagenome of an Acidic Hot Spring Microbial Planktonic Community in the Colombian Andes. PLoS ONE 7(12): e52069. doi:10.1371/journal.pone.0052069

Cyanobacterial diversity in the phyllosphere of a mangrove forest

The cyanobacterial community colonizing phyllosphere in a well-preserved Brazilian mangrove ecosystem was assessed using cultivation-independent molecular approaches. Leaves of trees that occupy this environment (Rhizophora mangle, Avicennia schaueriana and Laguncularia racemosa) were collected along a transect beginning at the margin of the bay and extending upland. The results demonstrated that the phyllosphere of R.similar to mangle and L.similar to racemosa harbor similar assemblages of cyanobacteria at each point along the transect. A.similar to schaueriana, found only in the coastal portions of the transect, was colonized by assemblages with lower richness than the other trees. However, the results indicated that spatial location was a stronger driver of cyanobacterial community composition than plant species. Distinct cyanobacterial communities were observed at each location along the coast-to-upland transect. Clone library analysis allowed identification of 19 genera of cyanobacteria and demonstrated the presence of several uncultivated taxa. A predominance of sequences affiliated with the orders Nostocales and Oscillatoriales was observed, with a remarkable number of sequences similar to genera Symphyonemopsis/Brasilonema (order Nostocales). The results demonstrated that phyllosphere cyanobacteria in this mangrove forest ecosystem are influenced by environmental conditions as the primary driver at the ecosystem scale, with tree species exerting some effect on community structure at the local scale.

Verrucosispora maris sp nov., a novel deep-sea actinomycete isolated from a marine sediment which produces abyssomicins

Verrucosispora isolate AB-18-032(T), the abyssomicin- and proximicin-producing actinomycete, has chemotaxonomic and morphological properties consistent with its classification in the genus Verrucosispora. The organism formed a distinct phyletic line in the Verrucosispora 16S rRNA gene tree sharing similarities of 99.7%, 98.7% and 98.9% with Verrucosispora gifhornensis DSM 44337(T), Verrucosispora lutea YIM 013(T) and Verrucosispora sediminis MS 426(T), respectively. It was readily distinguished from the two latter species using a range of phenotypic features and from V. gifhornensis DSM 44337(T), its nearest phylogenetic neighbor, by a DNA G+C content of 65.5 mol% obtained by thermal denaturation and fluorometry and DNA:DNA relatedness values of 64.0% and 65.0% using renaturation and fluorometric methods, respectively. It is apparent from the combined genotypic and phenotypic data that strain AB-18-032(T) should be classified in the genus Verrucosispora as a new species. The name Verrucosispora maris sp. nov. is proposed for this taxon with isolate AB-18-032(T) (= DSM 45365(T) = NRRL B-24793(T)) as the type strain.