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The cyanobacterial genus Nostoc is an important contributor to carbon and nitrogen bioavailability in terrestrial ecosystems and a frequent partner in symbiotic relationships with non-diazotrophic organisms. However, since this currently is a polyphyletic genus, the diversity of Nostoc -like cyanobacteria is considerably underestimated at this moment. While reviewing the phylogenetic placement of previously isolated Nostoc -like cyanobacteria originating from Brazilian Amazon, Caatinga and Atlantic forest samples, we detected 17 strains isolated from soil, freshwater, rock and tree surfaces presenting patterns that diverged significantly from related strains when ecological, morphological, molecular and genomic traits were also considered. These observations led to the identification of the evaluated strains as representative of three novel nostocacean genera and species: Amazonocrinis nigriterrae gen. nov., sp. nov.; Atlanticothrix silvestris gen. nov., sp. nov.; and Dendronalium phyllosphericum gen. nov., sp. nov., which are herein described according to the rules of the International Code of Nomenclature for algae, fungi and plants. This finding highlights the great importance of tropical and equatorial South American ecosystems for harbouring an unknown microbial diversity in the face of the anthropogenic threats with which they increasingly struggle.

The saline-alkaline lakes (soda lakes) are the habitat of the haloalkaliphilic cyanobacterium Anabaenopsis elenkinii, the type species of this genus. To obtain robust phylogeny of this type species, we have generated whole-genome sequencing of the bloom-forming Anabaenopsis elenkinii strain CCIBt3563 isolated from a Brazilian soda lake. This strain presents the typical morphology of A. elenkinii with short and curved trichomes with apical heterocytes established after separation of paired intercalary heterocytes and also regarding to cell dimensions. Its genome size is 4 495 068 bp, with a G+C content of 41.98 %, a total of 3932 potential protein coding genes and four 16S rRNA genes. Phylogenomic tree inferred by RAxML based on the alignment of 120 conserved proteins using GTDB-Tk grouped A. elenkinii CCIBt3563 together with other genera of the family Aphanizomenonaceae. However, the only previous available genome of Anabaenopsis circularis NIES-21 was distantly positioned within a clade of Desikacharya strains, a genus from the family Nostocaceae. Furthermore, average nucleotide identity values from 86–98 % were obtained among NIES-21 and Desikacharya genomes, while this value was 76.04 % between NIES-21 and the CCIBt3563 genome. These findings were also corroborated by the phylogenetic tree of 16S rRNA gene sequences, which also showed a strongly supported subcluster of A. elenkinii strains from Brazilian, Mexican and Kenyan soda lakes. This study presents the phylogenomics and genome-scale analyses of an Anabaenopsis elenkinii strain, improving molecular basis for demarcation of this species and framework for the classification of cyanobacteria based on the polyphasic approach.

Made available in DSpace on 2018-12-11T17:34:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-09-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Tropical ecosystems worldwide host very diverse microbial communities, but are increasingly threatened by deforestation and climate change. Thus, characterization of biodiversity in these environments, and especially of microbial communities that show unique adaptations to their habitats, is a very urgent matter. Information about representatives of the phylum Cyanobacteria in tropical environments is scarce, even though they are fundamental primary producers that help other microbes to thrive in nutrient-depleted habitats, including phyllospheres. In order to increase our knowledge of cyanobacterial diversity, a study was conducted to characterize isolates from Avicennia schaueriana and Merostachys neesii leaves collected at a mangrove and an Atlantic forest reserve located at the littoral of São Paulo state, south-east Brazil. The morphological, ultrastructural, phylogenetic, molecular and ecological features of the strains led to the recognition of the new genus Kryptousia, comprising two new species, Kryptousia macronema gen. nov., sp. nov. and Kryptousia microlepis sp. nov., described here according to the International Code of Nomenclature for algae, fungi and plants. The new genus and species were classified in the nostocalean family Tolypotrichaceae. This finding advances knowledge on the microbial diversity of South American ecosystems and sheds further light on the systematics of cyanobacteria. University of São Paulo USP Center for Nuclear Energy in Agriculture, Avenida Centenário 303 São Paulo State University UNESP Institute of Biosciences Humanities and Exact Sciences, Rua Cristóvão Colombo 2265 São Paulo State University UNESP Institute of Biosciences Humanities and Exact Sciences, Rua Cristóvão Colombo 2265 CNPq: #132494/2010-8 CNPq: #2008/52556-4 FAPESP: #2009/15402-1 CNPq: #310244/2015-3

Blooms of the toxic cyanobacterium Nodularia spumigena occur in various locations worldwide, but have not been observed in Brazil until recently. Three Nodularia strains were isolated from summer blooms in experimental shrimp production ponds of Penaeus vannamei in Rio Grande, in southern Brazil; these strains were characterized by morphology, phylogeny, growth rate and toxicity. The strains were identified as N. spumigena based on the size of vegetative cells, heterocytes and akinetes under a light microscope and based on the number of gas vesicles per μm2 under a transmission electron microscope. The 16S rRNA gene sequences of the three strains showed high identity (> 99%) with N. spumigena sequences available on the NCBI database but were grouped closer in the phylogenetic tree with N. spumigena strains from Australia and USA than those from the Baltic Sea. The growth rate in batch culture varied between 0.2 and 0.6 μ d−1 based on cell density, optical density and chlorophyll-a content. The thr...

Some Arcobacter species are considered emerging food-borne pathogens. However, their prevalence in dairy products is not well known. Therefore, the aim of the current research is to know the occurrence of Arcobacter species in Minas frescal cheese and raw cow milk and its potential association with physicochemical and microbiological parameters. Two hundred and one food samples (cheese, n = 98 and milk, n = 103) collected between 2016 and 2017 in Sao Paulo State, Brazil, were analyzed. Arcobacter was isolated using an enrichment broth with further filtration over Arcobacter agar. Colonies were characterized by ERIC-PCR, m-PCR and PCR-RFLP and, when was necessary, sequencing of 16S rRNA and rpoB genes were also assessed. Additionally, cheese samples were evaluated in relation to total coliform (TC), thermotolerant coliforms (TtC), coagulase positive staphylococci (CoPS) count and moisture. Raw milk samples were assessed for aerobic mesophilic bacteria count, TC, TtC, Dornic acidity, proteins, fats, lactose, defatted dry extract, total solids and somatic cell counts. The occurrence of Arcobacter was 10.2% (10/98) and 16.5% (17/103) in cheese and milk samples, respectively. All isolates were identified as Arcobacter butzleri. On the other hand, Multiple Factorial Analysis showed correlation between the presence of this species and the cheese moisture, while this pathogen was correlated with high counts of mesophilic microorganisms and TC in raw milk. No other evaluated parameter in cheese (TC, CoPS) and milk (TtC, Dornic acidity, proteins, fats, lactose, defatted dry extract, total solids and somatic cell counts) showed significant correlation. To date, this is the first report on A. butzleri in Minas frescal cheese, therefore its presence in dairy products could be considered a potential public health concern.

The taxonomy of four isolated strains of Anabaenopis elenkinii and A. arnoldii from North and South America were investigated using a polyphasic approach. The studied populations were collected from seven alkaline lakes of the Brazilian Pantanal and Nabor Carrillo Lake, Texcoco region (Mexico), between 2004 and 2012, where they frequently formed blooms during the dry season. The morphological plasticity of the populations, from natural and culture conditions, was described based on 200 individuals of each species. The 16S rRNA and the cpc-BA-IGS phylogenetic analyses show Anabaenopsis as a sister-taxon of Cyanospira . The Anabaenopsis clade has three strongly supported subclusters related to the origin of strains: Pantanal+Mexico, Kenya+Australia and one European clade. Although these data indicate that the currently known Anabaenopsis strains belong to more than one species, we are being conservative and not describing them as new species due to the lack of more molecular data. We highlight the need of a revision of the genus, considering also that Anabaenopsis sp. PCC 9215, the current reference strain used for the genus, is not identified as A. elenkinii and it is phylogenetically grouped with Anabaenopsis strains identified as A. circularis , A. arnoldii , A. nadsonii and A. elenkinii .

International audience; Soda lakes have high levels of sodium carbonates and are characterized by salinity and elevated pH. These ecosystems are found across Africa, Europe, Asia, Australia, North, Central, and South America. Particularly in Brazil, the Pantanal region has a series of hundreds of shallow soda lakes (ca. 600) potentially colonized by a diverse haloalkaliphilic microbial community. Biological information of these systems is still elusive, in particular data on the description of the main taxa involved in the biogeochemical cycling of life-important elements. Here, we used metagenomic sequencing to contrast the composition and functional patterns of the microbial communities of two distinct soda lakes from the sub-region Nhecolândia, state of Mato Grosso do Sul, Brazil. These two lakes differ by permanent cyanobacterial blooms (Salina Verde, green-water lake) and by no record of cyanobacterial blooms (Salina Preta, black-water lake). The dominant bacterial species in the Salina Verde bloom was Anabaenopsis elenkinii. This cyanobacterium altered local abiotic parameters such as pH, turbidity, and dissolved oxygen and consequently the overall structure of the microbial community. In Salina Preta, the microbial community had a more structured taxonomic profile. Therefore, the distribution of metabolic functions in Salina Preta community encompassed a large number of taxa, whereas, in Salina Verde, the functional potential was restrained across a specific set of taxa. Distinct signatures in the abundance of genes associated with the cycling of carbon, nitrogen, and sulfur were found. Interestingly, genes linked to arsenic resistance metabolism were present at higher abundance in Salina Verde and they were associated with the cyanobacterial bloom. Collectively, this study advances fundamental knowledge on the composition and genetic potential of microbial communities inhabiting tropical soda lakes.

Cyanobacteria from underexplored and extreme habitats are attracting increasing attention in the search for new bioactive substances. However, cyanobacterial communities from tropical and subtropical regions are still largely unknown, especially with respect to metabolite production. Among the structurally diverse secondary metabolites produced by these organisms, peptides are by far the most frequently described structures. In this work, liquid chromatography/electrospray ionization coupled to high resolution quadrupole time-of-flight tandem mass spectrometry with positive ion detection was applied to study the peptide profile of a group of cyanobacteria isolated from the Southeastern Brazilian coastal forest. A total of 38 peptides belonging to three different families (anabaenopeptins, aeruginosins, and cyanopeptolins) were detected in the extracts. Of the 38 peptides, 37 were detected here for the first time. New structural features were proposed based on mass accuracy data and isotopic patterns derived from full scan and MS/MS spectra. Interestingly, of the 40 surveyed strains only nine were confirmed to be peptide producers, all of these strains belonged to the order Nostocales (three Nostoc sp., two Desmonostoc sp. and four Brasilonema sp.).

Cyanobacteria are photosynthetic prokaryotes found in a range of environments. They are infamous for the production of toxins, as well as bioactive compounds, which exhibit anticancer, antimicrobial and protease inhibition activities. Cyanobacteria produce a broad range of antifungals belonging to structural classes, such as peptides, polyketides and alkaloids. Here, we tested cyanobacteria from a wide variety of environments for antifungal activity. The potent antifungal macrolide scytophycin was detected in Anabaena sp. HAN21/1, Anabaena cf. cylindrica PH133, Nostoc sp. HAN11/1 and Scytonema sp. HAN3/2. To our knowledge, this is the first description of Anabaena strains that produce scytophycins. We detected antifungal glycolipopeptide hassallidin production in Anabaena spp. BIR JV1 and HAN7/1 and in Nostoc spp. 6sf Calc and CENA 219. These strains were isolated from brackish and freshwater samples collected in Brazil, the Czech Republic and Finland. In addition, three cyanobacterial strains, Fischerella sp. CENA 298, Scytonema hofmanni PCC 7110 and Nostoc sp. N107.3, produced unidentified antifungal compounds that warrant further characterization. Interestingly, all of the strains shown to produce antifungal compounds in this study belong to Nostocales or Stigonematales cyanobacterial orders.

Cyanobacterial communities on the phyllosphere of 4 plant species inhabiting the endangered Brazilian Atlantic Forest biome were evaluated using cultivation-independent molecular approaches. Total genomic DNA was extracted from cells detached from the surface of leaves of Euterpe edulis, Guapira opposita, Garcinia gardneriana, and Merostachys neesii sampled in 2 Brazilian Atlantic Forest locations along an elevational gradient, i.e., lowland and montane forest. The DNA fingerprinting method PCR–DGGE revealed that the cyanobacterial phyllosphere community structures were mainly influenced by the plant species; geographical location of the plant had little effect. The 16S rRNA gene sequences obtained by clone libraries showed a predominance of nitrogen-fixing cyanobacteria of the order Nostocales, even though the majority of retrieved operational taxonomic units (∼60% of the sequences) showed similarity only to uncultured cyanobacteria phylotypes. The leaf surface of Guapira opposita had the highest richness and diversity of cyanobacteria, whereas the M. neesii (bamboo) had the largest number of copies of cyanobacterial 16S rRNA gene per cm2of leaf. This study investigated cyanobacteria diversity and its distribution pattern in Atlantic forest phyllosphere. The results indicated that plant species is the main driver of cyanobacteria community assemblage in the phyllosphere and that these communities are made up of a high diversity of cyanobacterial taxa that need to be discovered.

Studies investigating the diversity of cyanobacteria from tropical environments are scarce, especially those devoted to the isolation and molecular characterization of the isolated strains. Among the Brazilian biomes, Pantanal has mainly been examined through microscopic observation of environmental samples, resulting in lists of morphotypes without any genetic information. Recently, two studies were conducted evaluating the morphologic and genetic diversity of cultured non-heterocytous cyanobacteria in this biome, which resulted in the separation and description of two novel genera. In order to complement the diversity of cultured cyanobacteria from saline-alkaline lakes in Pantanal, the present study is dedicated to the examination of cultured nitrogen-fixing heterocytous cyanobacteria from this extreme and underexplored environment. A total of fourteen cyanobacterial strains were isolated. According to morphological examination they belong to the order Nostocales and to the subsections IV.I and IV.II, according to the International Code of Nomenclature for Algae, Fungi and Plants and the Bergey’s Manual of Systematic Bacteriology, respectively. Phylogenetic evaluation of their 16S rRNA gene sequences resulted in the formation of five clusters. Among them, one is clearly related to the genus Anabaenopsis whilst the remaining clusters may represent new genetic lineages. These novel sequences aid in the delimitation of problematic groups, especially those containing sequences belonging to mixed genera. The application of both morphologic and phylogenetic studies has proven to be an important tool in resolving problematic groups in cyanobacteria systematics. This strategy is essential in order to detect novel cyanobacteria genera from other tropical environments.

We report here the draft genome assembly of the brackish cyanobacterium Nodularia spumigena strain CENA596 isolated from a shrimp production pond in Rio Grande do Sul, Brazil. The draft genome consists of 291 contigs with a total size of 5,189,679 bp. Secondary metabolite annotations resulted in several predicted gene clusters, including those responsible for encoding the hepatotoxin nodularin.

Os microRNAs (miRNAs) são pequenos RNAs não codificadores, de cerca de 20 bases de comprimento, capazes de regular negativamente a expressão gênica após a transcrição. A ação regulatória destas moléculas é indispensável para o funcionamento adequado de diversos processos biológicos, dentre eles o desenvolvimento e a manutenção da musculatura esquelética. Com o objetivo de caracterizar a população de miRNAs presentes na musculatura esquelética adulta de frangos, foi construída uma biblioteca de miRNAs a partir do músculo peitoral de indivíduos jovens (21 dias pós-eclosão). Um total de 576 clones foi sequenciado e as sequências obtidas foram agrupadas por similaridade em 98 grupos, dentre os quais 47 corresponderam à miRNAs conhecidos, 30 à outros tipos de RNA e seis à possíveis novos miRNAs. Estes dados poderão subsidiar estudos funcionais subsequentes, que visem entender a função exercida por cada uma destas moléculas na musculatura esquelética. Buscando-se associar a ocorrência e expressão dos miRNAs ao controle da miogênese, foi analisada a expressão de três miRNAs identificados na biblioteca (miR-125b, miR-221 e miR-206), envolvidos na regulação do balanço entre proliferação e diferenciação celular, mecanismo determinante da miogênese. A análise foi realizada por PCR quantitativa em diferentes estádios do desenvolvimento (nove e 17 dias embrionário e 21 dias pós-eclosão) e entre duas linhagens de frango com potencial divergente para crescimento e ganho de massa muscular. Não houve diferença significativa na expressão dos miRNAs entre as linhagens em nenhum dos estádios aferidos, entretanto, foi possível traçar a ontogenia destas moléculas ao longo do desenvolvimento do animal, o que permitiu inferências sobre as condições morfológicas e fisiológicas das células musculares em cada um dos estádios analisados. Por fim, com o objetivo de associar os dados de expressão obtidos para os miRNAs, à variações na expressão de genes alvo, aferimos a expressão de três genes: SRF, Fstl e Pola1; onde o primeiro é regulado pelo miR-133a (cuja expressão não foi aferida devido à questões metodológicas) e os dois últimos pelo miR-206. A análise foi feita também por PCR quantitativa, entre as linhagens e em diferentes estádios do desenvolvimento. Foi possível visualizar, apenas nos estádios embrionários, a relação entre a expressão do miR-206 e seus genes alvo, com uma coerência entre o aumento na expressão do miR-206 e a diminuição na expressão de Pola1 e Fstl1. A determinação do perfil de expressão dos três genes ao longo do desenvolvimento muscular permitiu inferências sobre a ação destas moléculas no balanço entre proliferação e diferenciação nas linhagens de corte e postura MicroRNAs (miRNAs) represent a class of small and noncoding RNAs, about 20-nucleotides long that negatively regulate gene expression posttranscriptionally. The regulatory action of these molecules is essential for the proper functioning of various biological processes, including the development and maintenance of skeletal muscles. To identify miRNAs that might be important for the skeletal muscle development, we constructed a miRNA library from pectoral skeletal muscle of 21º days after birth chickens. A total of 576 clones were sequenced and these sequences were collapsed into 98 clusters. Sequence analysis identified 47 small RNAs that show significant similarities with published miRNAs, 30 with others noncoding RNAs and six sequences clusters could be identified as potentially novel miRNAs. These data may support subsequent functional studies aimed at understanding the function performed by each of these molecules in skeletal muscle of chicken. To further associate the miRNA presence with the gene expression in the controlling of myogenesis the expression patterns of tree miRNAs identified in the library (miR-125b, miR-221 e miR-206) were analyzed. These miRNAs are involved in the balance between proliferation and differentiation mechanisms that control myogenesis. The expressions of these miRNAs were measured using quantitative RT-PCR. We analyzed samples from two embryos stages (9 and 17 days) and one adult stage (21º days after birth) in two chicken lines with different potential to growth and gain of muscle mass. There were no significant differences between the lines about these miRNAs expression. But, we could predict an overview of these miRNAs expressions during the muscle development of chicken, which allowed inferences about the physiologic and morphologic conditions of the muscles cells in each analyzed stage. Also, to further associate the miRNAs results to variations in the target genes expressions, we analyzed the expression of three genes: SRF, Fstl e Pola1. The first one is target of miR-133a (not analyzed due to methodological problems), and the others are target of miR-206. We analyzed by quantitative RT-PCR different stages and two chicken lines. It was possible to observe, only in the earlier stages, a relationship between the miR-206 expression and the Pola1 and Fstl1 expression, with a consistency between the increased expression of mir-206 and decrease in expression of Pola1 and Fstl1. The determination of the profile of these three gene expressions during the muscle development allowed inferences about the action of these molecules in the balance between proliferation and differentiation process in chicken strains for broiler and layer

The genus Leptolyngbya Anagnostidis & Komárek (1988) was described from a set of strains identified as ‘LPP-group B’. The morphology within this group is not particularly informative and underestimates the group’s genetic diversity. In the present study, two new pseudanabaenacean genera related to Leptolyngbya morphotypes, Pantanalinema gen. nov. and Alkalinema gen. nov., are described under the provisions of the International Code of Nomenclature for Algae, Fungi and Plants, based on a polyphasic approach. Pantanalinema gen. nov. (type species Pantanalinema rosaneae sp. nov.) has sheaths and trichomes with slight gliding motility, which distinguish this genus from Alkalinema gen. nov. (type species Alkalinema pantanalense sp. nov.), which possesses trichomes arranged in an ornate (interwoven) pattern. 16S rRNA gene sequences of strains of Pantanalinema and Alkalinema exhibited low identity to each other (≤91.6 %) and to other sequences from known pseudanabaenacean genera (≤94.3 and 93.7 %, respectively). In a phylogenetic reconstruction, six sequences from strains of Pantanalinema and four from strains of Alkalinema formed two separate and robust clades (99 % bootstrap value), with the genera Oculatella and Phormidesmis, respectively, as the closest related groups. 16S–23S rRNA intergenic spacer sequences and secondary structures of strains of Pantanalinema and Alkalinema did not correspond to any previous descriptions. The strains of Pantanalinema and Alkalinema were able to survive and produce biomass at a range of pH (pH 4–11) and were also able to alter the culture medium to pH values ranging from pH 8.4 to 9.9. These data indicate that cyanobacterial communities in underexplored environments, such as the Pantanal wetlands, are promising sources of novel taxa.

Cyanobacteria possess the ability to produce compounds with remarkable biological activity, and have thus attracted the attention of the pharmaceutical industry. Cyanopeptides acting as protease inhibitors have shown potential in the field of pharmacotherapy through regulation of abnormal physiological processes in the human body. Despite the already described cyanopeptide protease inhibitors, the search for new congeners is of considerable interest which may pave the way for more efficient molecules. In this study, the presence of the protease inhibitors aeruginosin and cyanopeptolin with non-, mono- and dichlorination and also genes coding for their synthetases was investigated in 90 cyanobacterial strains. Mass spectrometry analyses highlighted production of 91, 19 and 3 non-, mono- and dichlorinated congeners, respectively. The purified extract of Microcystis botrys SPC759 inhibited 61% of pepsin protease. PCR amplifications of aeruginosin and cyanopeptolin synthetase gene regions were observed in 41 and 28% of evaluated strains, respectively. The sequences obtained for the aerA-aerB (aeruginosin) and mcnC-mcnE (cyanopeptolin) gene regions grouped together with their homologues found in other cyanobacterial strains in the phylogenetic analyses with high bootstrap support. Antimicrobial activity assays performed using all intracellular extracts inhibited 31 and 26% of Gram-negative and Gram-positive pathogenic bacterial growth, respectively. The results of this study showed the production of aeruginosin and cyanopeptolin and the presence of their genes in several cyanobacterial genera for the first time besides the discovery of novel congeners.

O manejo agrícola aplicado a um agrossistema pode determinar a qualidade e produtividade da área, além das interações biológicas que podem ser estabelecidas entre o cultivar e ecossistema local. A agricultura convencional é bastante reconhecida como um manejo eficiente e lucrativo. Por outro lado, a agroecologia tem ganhado visibilidade na agroindústria em reflexo do aumento na demanda por alternativas mais sustentáveis de produção. As diferenças entre ambos manejos podem refletir sobre a dinâmica microbiana, alterando a composição e estruturação das comunidades ali presentes. Os micro-organismos que habitam a superfície foliar da planta compõem o micro-ambiente denominado filosfera, descrito como um dos hábitats colonizáveis mais extensos. Devido a sua alta exposição a variáveis ambientais, diversos fatores podem interferir na comunidade bacteriana e definir a filosfera. Desta forma, o principal objetivo nesse estudo foi avaliar como o manejo agrícola interfere na composição bacteriana na filosfera, analisando ainda em escala temporal sua estrutura e abundância. A área experimental amostrada foi cedida pelo Centro de Pesquisa \"Mokiti Okada\", em Mogi Guaçu, São Paulo. As amostras foram coletadas de maneira representativa em diferentes linhas de tratamento, uma sob manejo convencional e outra sob manejo ecológico. Análises microbiológicas dependentes e independentes de cultivo permitiram identificar a comunidade bacteriana residente da filosfera de citros, a qual era compartilhada por ambos os manejos. Entretanto, análises de sequenciamento NGS (New Generation Sequencing) mostraram uma diferença significativa entre as comunidades bacterianas dos dois manejos, com o ecológico apresentando uma maior diversidade. Apesar do manejo ter se mostrado um importante fator na composição bacteriana, quando avaliado em função temporal, viu-se que as épocas de coleta interferem mais intensamente na estrutura das bactérias (p=0,0001), mostrando uma sobreposição dos diversos fatores ambientais que atuam sobre a filosfera. Os resultados ainda indicaram uma redução na abundância de bactérias, a qual pode estar relacionada com a aplicação extra de produtos cúpricos em ambas as áreas, em função do acometimento da \"pinta preta\" no pomar, o que instigou monitorar o cobre no tecido foliar. Quimicamente, micro-análises de XRF (X-Ray Fluorescence) mostraram que há uma maior concentração de cobre nas folhas provenientes da área convencional, o que é resultado das maiores quantidades do produto que são aplicadas nesse tratamento. Além disso, foi possível o isolamento de bactérias do gênero Enterococcus na filosfera, as quais apresentam mecanismos de tolerância ao cobre, demonstrando que os produtos cúpricos podem ter selecionado esses organismos. Logo, esse estudo apresentou uma importante perspectiva do efeito do manejo agrícola sobre a filosfera, contribuindo para a compreensão da dinâmica microbiana na agricultura. The agricultural management applied to a agrosystem is an important determinant for the quality and productivity of the crop yield, also for the biological interactions that can be stablished between the plants and the local ecosystem. Conventional agriculture has being well known as an efficient and lucrative crop management. On the other hand, agroecology has gaining visibility in the agroindustry due to increasing demand for a more sustainable production alternative. The differences between both approaches can reflect on the microbial dynamic, affecting the composition and structure of these communities. Microrganisms inhabitating the foliar surface correspond to a microenvironment called phyllosphere, which is described as one of the most extensive habitats. Due to its constant exposition to environmental variables, several factors can influence on the bacterial community and modulate the phyllosphere. Thus, the main purpose of this study was to evaluate how the agricultural management can impact on the phyllospheric bacteria, also considering a temporal effect on structure and abundance of these organisms. The experimental area was provided by \"Mokiti Okada\" Research Center, located at em Mogi-Guaçu, São Paulo. The samples were representativelly collected from two treatment lines, one under convencional management, and the other under ecological management. Afterwards, culture-dependent and independent microbiological analysis allowed to identify the resident bacterial community in citros phyllosphere, which was greatly shared bewteen both treatments. However, NSG (New Generation Sequencing) analysis demonstrated a significative difference between the bacterial community under conventional and ecological management, where the second one demonstrated a higher diversity, which can be related to the different approaches applied. Although the agricultural method have demonstrated an important factor on bacterial composition, when temporally evaluated, it was observed a more intense interferance on the bacterial structure by the time of sampling (p=0,0001), representing a possible overlap of environmental factors on the phyllosphere. The data also indicate a decrease in the abundance of bacteria that might be resulted from the extra use of cupric products, related to the impairment of \"black spot\" on the crop, what lead to a copper monitoring in the foliar tissue. Chemically, XRF micro-analysis (X-Ray Fluorescence) demonstrated that there is a higher concentration of copper on the leaves from the conventional area, which is resulted of the higher application of its products by this method. Moreover, a search for copper-tolerant microrganisms was conducted, and it was possible to isolate Enterococcus bacteria, which have copper tolerance mechanisms. This result implicate that the use of cupric products may have selected these microrganisms on citros phyllosphere. Therefore, this study presented an important perspective of how the agricultural management can influence the phyllosfere, which can contribute to undertand about the microbial dynamic and its roles on the agriculture.

MicroRNAs (miRNAs, miRs) encompass a class of small non-coding RNAs that often negatively regulate gene expression. miRNAs play an essential role in skeletal muscle, determining the proper development and maintenance of this tissue. In comparison to other organs and tissues, the full set of muscle miRNAs and its expression patterns are still poorly understood. In this report, a chicken skeletal muscle miRNA library was constructed, and the expression of selected miRNAs was further characterized during muscle development in chicken lines with distinct muscling phenotypes. Clone library sequence analysis revealed 40 small RNAs with similarities to previously described chicken miRNAs, seven miRNAs that were never identified before in chicken, and some sequence clusters representing other possible novel miRNAs. Temporal expression profiles of three miRNAs associated with cell proliferation and differentiation (miR-125b, miR-221, and miR-206) in two chicken lines (broiler and layer) revealed the differential steady-state levels of these miRs during skeletal muscle growth and suggests that miR-206 is involved in the muscling phenotype that is observed in growth-selected chicken lines. Made available in DSpace on 2018-01-26T23:41:35Z (GMT). No. of bitstreams: 1 final7468.pdf: 657307 bytes, checksum: 637f9a8256d86f60213339f776fc4110 (MD5) Previous issue date: 2015-04-07