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THE HUMAN MICROBIOTA: THE ROLE OF MICROBIAL COMMUNITIES IN HEALTH AND DISEASE Acta biol.Colomb.
BOTERO,Luz Elena; DELGADO-SERRANO,Luisa; CEPEDA HERNÁNDEZ,Martha Lucía; DEL PORTILLO OBANDO,Patricia; ZAMBRANO EDER,María Mercedes.
During the last decade, there has been increasing awareness of the massive number of microorganisms, collectively known as the human microbiota, that are associated with humans. This microbiota outnumbers the host cells by approximately a factor of ten and contains a large repertoire of microbial genome-encoded metabolic processes. The diverse human microbiota and its associated metabolic potential can provide the host with novel functions that can influence host health and disease status in ways that still need to be analyzed. The microbiota varies with age, with features that depend on the body site, host lifestyle and health status. The challenge is therefore to identify and characterize these microbial communities and use this information to learn how...
Tipo: Info:eu-repo/semantics/article Palavras-chave: 16S rRNA gene; Microbial diversity; Microbiome; Mycobacterium tuberculosis.
Ano: 2016 URL: http://www.scielo.org.co/scielo.php?script=sci_arttext&pid=S0120-548X2016000100001
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Bacterial and Archaeal Communities Variability Associated with Upwelling and Anthropogenic Pressures in the Protection Area of Arraial do Cabo (Cabo Frio region - RJ) Anais da ABC (AABC)
COELHO-SOUZA,SERGIO A.; ARAÚJO,FÁBIO V.; CURY,JULIANO C.; JESUS,HUGO E.; PEREIRA,GILBERTO C.; GUIMARÃES,JEAN R.D.; PEIXOTO,RAQUEL S.; DÁVILA,ALBERTO M.R.; ROSADO,ALEXANDRE S..
ABSTRACTUpwelling 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...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Microbial diversity; Leucine incorporation; Flow cytometry; Temperature; Nutrients availability; Brazilian Upwelling System.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652015000401737
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Microbiological, technological and therapeutic properties of kefir: a natural probiotic beverage BJM
Leite,Analy Machado de Oliveira; Miguel,Marco Antonio Lemos; Peixoto,Raquel Silva; Rosado,Alexandre Soares; Silva,Joab Trajano; Paschoalin,Vania Margaret Flosi.
Kefir is a fermented milk beverage produced by the action of bacteria and yeasts that exist in symbiotic association in kefir grains. The artisanal production of the kefir is based on the tradition of the peoples of Caucasus, which has spread to other parts of the world, from the late 19th century, and nowadays integrates its nutritional and therapeutic indications to the everyday food choices of several populations. The large number of microorganisms present in kefir and their microbial interactions, the possible bioactive compounds resulting of microbial metabolism, and the benefits associated with the use this beverage confers kefir the status of a natural probiotic, designated as the 21th century yoghurt. Several studies have shown that kefir and its...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Probiotic; Microbial diversity; Kefir grains; Fermented milk; Kefir.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822013000200001
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Microbial diversity in a bagasse-based compost prepared for the production of Agaricus brasiliensis BJM
Silva,Cristina Ferreira; Azevedo,Raquel Santos; Braga,Claudia; Silva,Romildo da; Dias,Eustáquio Souza; Schwan,Rosane Freitas.
Edible mushrooms are renowned for their nutritional and medicinal properties and are thus of considerable commercial importance. Mushroom production depends on the chemical composition of the basic substrates and additional supplements employed in the compost as well as on the method of composting. In order to minimise the cost of mushroom production, considerable interest has been shown in the use of agro-industrial residues in the preparation of alternative compost mixtures. However, the interaction of the natural microbiota present in agricultural residues during the composting process greatly influences the subsequent colonisation by the mushroom. The aim of the present study was to isolate and identify the microbiota present in a sugar cane bagasse...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Agaricus brasiliensis; Cane sugar bagasse; Microbial diversity.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822009000300023
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Distribution of prokaryotic organisms in a tropical estuary influenced by sugar cane agriculture in northeast Brazil BJM
Wolf,Lars; Schwalger,Berit; Knoppers,Bastiaan A.; Silva,Luiz Antonio Ferreira da; Medeiros,Paulo Ricardo Petter; Pollehne,Falk.
In a joint Brazilian-German case study, distribution patterns of microorganisms were compared with environmental variables in the tropical coastal Manguaba lagoon in northeast Brazil, which is situated downstream of several sugar cane processing plants . 16S rDNA and 16S rRNA single strand conformation polymorphism (SSCP) gene fingerprinting were used to follow the composition and distribution of microorganisms throughout the salinity gradient of the lagoon. Potentially abundant microorganisms were identified by sequencing representative SSCP bands. It could be demonstrated that the distribution of microbes was in close relation to the physico-chemical environmental settings and followed a common scheme. In the in- and outlet areas of the lagoon rather...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Tropical lagoon; Estuarine gradient; Microbial diversity; Sugar cane.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822010000400007
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Analysis of microbial diversity in Shenqu with different fermentation times by PCR-DGGE BJM
Liu,Tengfei; Jia,Tianzhu; Chen,Jiangning; Liu,Xiaoyu; Zhao,Minjie; Liu,Pengpeng.
Abstract Shenqu is a fermented product that is widely used in traditional Chinese medicine (TCM) to treat indigestion; however, the microbial strains in the fermentation process are still unknown. The aim of this study was to investigate microbial diversity in Shenqu using different fermentation time periods. DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) profiles indicated that a strain of Pediococcus acidilactici (band 9) is the predominant bacteria during fermentation and that the predominant fungi were uncultured Rhizopus, Aspergillus oryzae, and Rhizopus oryzae. In addition, pathogenic bacteria, such as Enterobacter cloacae, Klebsiella oxytoca, Erwinia billingiae, and Pantoea vagan were detected in Shenqu. DGGE analysis...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Microbial diversity; PCR-DGGE; Shenqu; Molecular cloning.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822017000200246
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Molecular characterization of bacterial populations of different soils BJM
Pereira,Rodrigo Matheus; Silveira,Érico Leandro da; Scaquitto,Denilson César; Pedrinho,Eliamar Aparecida Nascimbém; Val-Moraes,Silvana Pompéia; Wickert,Ester; Carareto-Alves,Lúcia Maria; Lemos,Eliana Gertrudes de Macedo.
Until recently, few studies were carried out in Brazil about diversity of bacterial soil communities. Aiming to characterize the bacterial population in the soil through 16S rRNA analysis, two types of soil have been analyzed: one of them characterized by intensive use where tomato, beans and corn were cultivated (CS); the other analyzed soil was under forest (FS), unchanged by man; both located in Guaíra, São Paulo State, Brazil. Using specific primers, 16S rRNA genes from metagenomic DNA in both soils were amplified by PCR, amplicons were cloned and 139 clones from two libraries were partially sequenced. The use of 16S rRNA analysis allowed identification of several bacterial populations in the soil belonging to the following phyla: Acidobacteria,...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Metagenome; Microbial diversity; 16S rRNA.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822006000400007
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Comparison of different protocols for the extraction of microbial DNA from reef corals BJM
Santos,H.F.; Carmo,F.L.; Leite,D.C.A.; Jesus,H.E.; De Carvalho Maalouf,P.; Almeida,C.; Soriano,A.U.; Altomari,D.; Suhett,L.; Vólaro,V.; Valoni,E.; Francisco,M.; Vieira,J.; Rocha,R.; Sardinha,B.L.; Mendes,L.B.; João,R.R.; Lacava,B.; Jesus,R.F.; Sebastian,G.V.; Pessoa,A.; van Elsas,J.D.; Rezende,R.P.; Pires,D.O.; Duarte,G.; Castro,C.B.; Rosado,A.S.; Peixoto,R.S..
This study aimed to test different protocols for the extraction of microbial DNA from the coral Mussismilia harttii. Four different commercial kits were tested, three of them based on methods for DNA extraction from soil (FastDNA SPIN Kit for soil, MP Bio, PowerSoil DNA Isolation Kit, MoBio, and ZR Soil Microbe DNA Kit, Zymo Research) and one kit for DNA extraction from plants (UltraClean Plant DNA Isolation Kit, MoBio). Five polyps of the same colony of M. harttii were macerated and aliquots were submitted to DNA extraction by the different kits. After extraction, the DNA was quantified and PCR-DGGE was used to study the molecular fingerprint of Bacteria and Eukarya. Among the four kits tested, the ZR Soil Microbe DNA Kit was the most efficient with...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Coral; Microbial diversity; DNA extraction.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822012000200012
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Characterization of spoilage bacteria in pork sausage by PCR-DGGE analysis Ciênc. Tecnol. Aliment.
Dias,Francesca Silva; Ramos,Cíntia Lacerda; Schwan,Rosane Freitas.
To investigate microbial diversity and identify spoilage bacteria in fresh pork sausages during storage, twelve industrial pork sausages of different trademarks were stored at 4 ºC for 0, 14, 28 and 42 days, 80% relative humidity and packaged in sterile plastic bags. Microbiological analysis was performed. The pH and water activity (a w) were measured. The culture-independent method performed was the Polymerase Chain Reaction - Denaturing Gradient Gel Electrophoresis (PCR-DGGE). The culture-dependent method showed that the populations of mesophilic bacteria and Lactic Acid Bacteria (LAB) increased linearly over storage time. At the end of the storage time, the average population of microorganisms was detected, in general, at the level of 5 log cfu g-1. A...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Microbial diversity; Lactobacillus sakei; Brochothrix thermosphacta.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612013000300013
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Microbial communities and their biogeochemical role in the water column of the oxygen minimum zone in the eastern South Pacific Gayana
Ulloa,Osvaldo; Belmar,Lucy; Farías,Laura; Castro-González,Maribel; Galán,Alexander; Lavín,Paris; Molina,Verónica; Ramírez,Salvador; Santibáñez,Francisco; Stevens,Heike.
In recent years, our group has been studying the microbial communities of the water column associated with the oxygen minimum zone (OMZ) off northern Chile and Peru, trying to understand who the main players are in the OMZ biogeochemical cycles and which metabolic strategies they are using. For that, we are combining flow cytometry, molecular techniques and biogeochemical approaches. Most of our work, so far, has focused on microbes capable of doing oxygenic photosynthesis, such as the cyanobacteria, and those involved in the nitrogen cycle, such as the denitrifying, nitrifying and anammox bacteria. We have also started to look at the more general microbial abundance and diversity. In each case, we are finding that the OMZ holds distinct microbial...
Tipo: Journal article Palavras-chave: Oxygen minimum zone; Microbial diversity; Nitrogen cycle.
Ano: 2006 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-65382006000300018
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Continuous enrichment cultures: insights into prokaryotic diversity and metabolic interactions in deep-sea vent chimneys ArchiMer
Postec, Anne; Lesongeur, Francoise; Pignet, Patricia; Ollivier, B; Querellou, Joel; Godfroy, Anne.
The prokaryotic diversity of culturable thermophilic communities of deep-sea hydrothermal chimneys was analysed using a continuous enrichment culture performed in a gas-lift bioreactor, and compared to classical batch enrichment cultures in vials. Cultures were conducted at 60 degrees C and pH 6.5 using a complex medium containing carbohydrates, peptides and sulphur, and inoculated with a sample of a hydrothermal black chimney collected at the Rainbow field, Mid-Atlantic Ridge, at 2,275 m depth. To assess the relevance of both culture methods, bacterial and archaeal diversity was studied using cloning and sequencing, DGGE, and whole-cell hybridisation of 16S rRNA genes. Sequences of heterotrophic microorganisms belonging to the genera Marinitoga,...
Tipo: Text Palavras-chave: Metabolic interactions; 16S rRNA gene; Thermophiles; Bioreactor; Continuous enrichment cultures; Deep sea hydrothermal vent; Microbial diversity.
Ano: 2007 URL: http://archimer.ifremer.fr/doc/2007/publication-3885.pdf
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Effect of variation of environmental conditions on the microbial communities of deep-sea vent chimneys, cultured in a bioreactor ArchiMer
Byrne, Nathalie; Lesongeur, Francoise; Bienvenu, Nadege; Geslin, Claire; Alain, Karine; Prieur, Daniel; Godfroy, Anne.
Both cultivation and molecular techniques were used to investigate the microbial diversity and dynamic of a deep-sea vent chimney. The enrichment cultures performed in a gas-lift bioreactor were inoculated with a black smoker chimney sample collected on TAG site on the mid-Atlantic ridge. To mimic as close as possible environmental conditions, the cultures were performed in oligotrophic medium with nitrogen, hydrogen and carbon dioxide (N-2/H-2/CO2) gas sweeping. Also, the temperature was first settled at a temperature of 85A degrees C and colloidal sulphur was added. Then, the temperature was lowered to 60A degrees C and sulphur was omitted. Archaeal and bacterial diversity was studied in both culture and natural samples. Through 16S rRNA gene sequences...
Tipo: Text Palavras-chave: 16S rRNA genes; Oligotrophic condition; Environmental condition; Bioreactor; Continuous enrichment culture; Microbial diversity; Deep sea hydrothermal vent.
Ano: 2009 URL: http://archimer.ifremer.fr/doc/2009/publication-6920.pdf
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Metagenomic and PCR-Based Diversity Surveys of [FeFe]-Hydrogenases Combined with Isolation of Alkaliphilic Hydrogen-Producing Bacteria from the Serpentinite-Hosted Prony Hydrothermal Field, New Caledonia ArchiMer
Mei, Nan; Postec, Anne; Monnin, Christophe; Pelletier, Bernard; Payri, Claude E.; Menez, Benedicte; Frouin, Eleonore; Ollivier, Bernard; Erauso, Gael; Quemeneur, Marianne.
High amounts of hydrogen are emitted in the serpentinite-hosted hydrothermal field of the Prony Bay (PHF, New Caledonia), where high-pH (similar to 11), low-temperature (< 40 degrees C), and low-salinity fluids are discharged in both intertidal and shallow submarine environments. In this study, we investigated the diversity and distribution of potentially hydrogen-producing bacteria in Prony hyperalkaline springs by using metagenomic analyses and different PCR-amplified DNA sequencing methods. The retrieved sequences of hydA genes, encoding the catalytic subunit of [FeFe]-hydrogenases and, used as a molecular marker of hydrogen-producing bacteria, were mainly related to those of Firmicutes and clustered into two distinct groups depending on sampling...
Tipo: Text Palavras-chave: Hydrogen; Microbial diversity; Hydrogen producers; Serpentinization; HydA genes; [FeFe]-hydrogenase; Metagenomics.
Ano: 2016 URL: https://archimer.ifremer.fr/doc/00467/57913/60308.pdf
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Comparative Study of Guaymas Basin Microbiomes: Cold Seeps vs. Hydrothermal Vents Sediments ArchiMer
Cruaud, Perrine; Vigneron, Adrien; Pignet, Patricia; Caprais, Jean-claude; Lesongeur, Francoise; Toffin, Laurent; Godfroy, Anne; Cambon-bonavita, Marie-anne.
In the Guaymas Basin, the presence at a few tens of kilometers of cold seeps and hydrothermal vents coupled with comparable sedimentary settings and depths offer a unique opportunity to assess and compare the microbial community composition of these deep-sea ecosystems. The microbial diversity in sediments from three cold seep and two hydrothermal vent assemblages were investigated using high-throughput 16S rRNA-sequencing. Numerous bacterial and archaeal lineages were detected in both cold seep and hydrothermal vent sediments. Various potential organic matter degraders (e.g., Chloroflexi, Atribacteria, MBG-D) and methane and sulfur cycling related microorganisms (e.g., ANME and methanogenic lineages, sulfate-reducing lineages) were detected in both...
Tipo: Text Palavras-chave: Microbial diversity; Hydrothermal vents; Cold seeps; Ribotypes; Methane; Extreme environments.
Ano: 2017 URL: https://archimer.ifremer.fr/doc/00415/52640/53496.pdf
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First insights into the microbiome of a carnivorous sponge ArchiMer
Dupont, Samuel; Corre, Erwan; Li, Yanyan; Vacelet, Jean; Bourguet-kondracki, Marie-lise.
Using 454 pyrosequencing, we characterized for the first time the associated microbial community of the deep-sea carnivorous Demosponge Asbestopluma hypogea (Cladorhizidae). Targeting the 16S rRNA gene V3 and V6 hypervariable regions, we compared the diversity and composition of associated microbes of two individual sponges of A.hypogea freshly collected in the cave with surrounding seawater and with one sponge sample maintained 1year in an aquarium after collection. With more than 22961 high quality sequences from sponge samples, representing c. 800 operational taxonomic units per sponge sample at 97% sequence similarities, the phylogenetic affiliation of A.hypogea-associated microbes was assigned to 20 bacterial and two archaeal phyla, distributed into...
Tipo: Text Palavras-chave: Asbestopluma hypogea; Carnivorous sponge; Cladorhizidae; Microbial diversity; Microscopic studies; Pyrosequencing 454 amplicons.
Ano: 2013 URL: https://archimer.ifremer.fr/doc/00372/48277/48534.pdf
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Phylogenetic characterization of the bacterial assemblage associated with mucous. secretions of the hydrothermal vent polychaete Paralvinella palmiformis ArchiMer
Alain, Karine; Olagnon, Michel; Desbruyeres, Daniel; Page, Antoine; Barbier, Georges; Juniper, Kim; Querellou, Joel; Cambon-bonavita, Marie-anne.
As part of an ongoing examination of microbial diversity associated with hydrothermal vent polychaetes of the family Alvinellidae, we undertook a culture-independent molecular analysis of the bacterial assemblage associated with mucous secretions of the Northeastern Pacific vent polychaete Paralvinella palmiformis. Using a molecular 16S rDNA-based phylogenetic approach, clone libraries were constructed from two samples collected from active sulfide edifices in two hydrothermal vent fields. In both cases, clone libraries were largely dominated by epsilon-Proteobacteria. Phylotypes belonging to the Cytophaga-Flavobacteria and to the Verrucomicrobia were also largely represented within the libraries. The remaining sequences were related to the taxonomic...
Tipo: Text Palavras-chave: Proteobacteria; 16S rRNA; Microbial diversity; Phylogeny; Hydrothermal vent; Paralvinella palmiformis.
Ano: 2002 URL: https://archimer.ifremer.fr/doc/2002/publication-661.pdf
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Role of environmental factors for the vertical distribution (0-1000 m) of marine bacterial communities in the NW Mediterranean Sea ArchiMer
Ghiglione, J. F.; Palacios, C.; Marty, J. C.; Mevel, G.; Labrune, C.; Conan, P.; Pujo-pay, M.; Garcia, N.; Goutx, M..
Bacterioplankton plays a central role in energy and matter fluxes in the sea, yet the factors that constrain its variation in marine systems are still poorly understood. Here we use the explanatory power of direct multivariate gradient analysis to evaluate the driving forces exerted by environmental parameters on bacterial community distribution in the water column. We gathered and analysed data from a one month sampling period from the surface to 1000m depth at the JGOFS-DYFAMED station (NW Mediterranean Sea). This station is characterized by very poor horizontal advection currents which makes it an ideal model to test hypotheses on the causes of vertical stratification of bacterial communities. Capillary electrophoresis single strand conformation...
Tipo: Text Palavras-chave: Free living bacteria; Ribosomal RNA genes; Strand conformation polymorphism; Flame ionization detection; Dissolved organic carbon; 16S RDNA; Microbial diversity; Lipid classes; Phytoplankton dynamics; Multivariate analyses.
Ano: 2008 URL: https://archimer.ifremer.fr/doc/00208/31974/30409.pdf
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Some considerations for analyzing biodiversity using integrative metagenomics and gene networks ArchiMer
Bittner, Lucie; Halary, Sebastien; Payri, Claude; Cruaud, Corinne; De Reviers, Bruno; Lopez, Philippe; Bapteste, Eric.
Background: Improving knowledge of biodiversity will benefit conservation biology, enhance bioremediation studies, and could lead to new medical treatments. However there is no standard approach to estimate and to compare the diversity of different environments, or to study its past, and possibly, future evolution. Presentation of the hypothesis: We argue that there are two conditions for significant progress in the identification and quantification of biodiversity. First, integrative metagenomic studies - aiming at the simultaneous examination (or even better at the integration) of observations about the elements, functions and evolutionary processes captured by the massive sequencing of multiple markers - should be preferred over DNA barcoding projects...
Tipo: Text Palavras-chave: Microbial diversity; Species concept; Dna barcodes; Ecological perspective; Community; Functional analysis; Rhodophyta; Systematics; Fragments; Taxonomy.
Ano: 2010 URL: https://archimer.ifremer.fr/doc/00202/31321/29733.pdf
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Microbial communities in Cerrado soils under native vegetation subjected to prescribed fire and under pasture PAB
Viana,Laura Tillmann; Bustamante,Mercedes Maria da Cunha; Molina,Marirosa; Pinto,Alexandre de Siqueira; Kisselle,Keith; Zepp,Richard; Burke,Roger A.
The objective of this work was to evaluate the effects of fire regimes and vegetation cover on the structure and dynamics of soil microbial communities, through phospholipid fatty acid (PLFA) analysis. Comparisons were made between native areas with different woody covers ("cerrado stricto sensu" and "campo sujo"), under different fire regimes, and a 20-year-old active palisadegrass pasture in the Central Plateau of Brazil. Microbial biomass was higher in the native plots than in the pasture, and the highest monthly values were observed during the rainy season in the native plots. No significant differences were observed between fire regimes or between communities from the two native vegetation types. However, the principal component (PC) analysis...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Land use; Microbial diversity; PLFA; Savannas; Seasonality.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-204X2011001200012
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The Effect of Hydrated Sodium Calcium Aluminosilicate on Fatty Liver and the Composition of the Intestinal Microbiota in Overfed Landes Geese Rev. Bras. Ciênc. Avic.
Tang,J; Fang,Q; Lu,M; Shao,R; Shen,J; Lu,L; Niu,D.
ABSTRACT Goose fatty liver is a delicious food product and the overfeeding will cause the abnormal physiology of the geese. The objective of this study was to investigate the effect of supplementation with hydrated sodium calcium aluminosilicate (HSCAS) on the fatty liver, ileal and cecal microbiota of Landes geese during overfeeding. Sixty 70-day-old Landes geese (body weight= 3.0 ± 0.05 kg) were randomly divided into three groups, two of which were overfed with whole corn supplemented with or without HSCAS for 20 days when the fatty liver reaches to the maximum size and the negative control group was ad libitum access to the corn basal diet. The intestinal contents of the ileum and cecum from three geese per group were used for high-throughput...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Landes geese; Microbial diversity; Overfeeding; Hydrated sodium calcium aluminosilicate; High-throughput sequencing.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2018000200393
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