Microbiologia
URI permanente desta comunidadehttps://locus.ufv.br/handle/123456789/11840
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Resultados da Pesquisa
Item Expression of the nifH gene in diazotrophic bacteria in Eucalyptus urograndis plantations(Canadian Journal of Forest Research, 2016-02) Silva, Marliane de Cássia Soares da; Mendes, Igor Rodrigues; Paula, Thiago de Almeida; Dias, Roberto Sousa; Paula, Sérgio Oliveira de; Silva, Cynthia Canedo; Bazzolli, Denise Mara Soares; Kasuya, Maria Catarina MegumiA large proportion of eucalypt plantations in Brazil are located in areas with low soil fertility. The actions of microorganisms are of great importance for the cycling of nutrients, including nitrogen (N), that are essential for plant metabolism. Denaturing gradient gel electrophoresis (DGGE) was used to monitor and identify the total and active microorganisms involved in the N cycle in both the soil and root systems of a forest of Eucalyptus urograndis with sections that were fertilized with N or unfertilized. Quantitative real-time PCR was used to examine the expression of the nifH gene in N-fixing bacteria present in both the soil and root systems. According to the DGGE analysis, in the total and active populations of N-fixing bacteria, the presence and expression of the nifH gene were influenced by the winter and summer seasons and (or) N fertilization, respectively. DGGE band sequencing from total DNA samples showed that the most abundant group of diazotrophic bacteria belonged to Alphaproteobacteria in both the soil and root systems. Quantitative real-time PCR revealed that nifH expression was higher in the soil samples, especially in those that did not receive N fertilization. The differences in the composition of the total and active diazotrophic populations highlight the importance of evaluating the active populations, because they are effectively responsible for the biogeochemical transformation of N and also control its' availability to plants.Item Morphological and molecular characterization of Tulasnella spp. fungi isolated from the roots of Epidendrum secundum, a widespread Brazilian orchid(Symbiosis, 2014-02) Pereira, Marlon Corrêa; Coelho, Irene da Silva; Valadares, Rafael Borges da Silva; Oliveira, Sabrina Feliciano; Bocayuva, Melissa; Pereira, Olinto Liparini; Araújo, Elza Fernandes; Kasuya, Maria Catarina MegumiTulasnella spp. are the main fungal symbionts of Brazilian Epidendrum orchids. The taxonomy of these fungi is largely based on ITS rDNA similarity, but culture dependent techniques are still essential to establish the true biological entity of the mycobiont. The aim of this study was to characterize morphologically and molecularly 16 Tulasnella spp. fungi isolated from three different populations of E. secundum and to test the coincidences between morphological and molecular characterization. Two uninucleate rhizoctonia fungi, obtained from Oncidium barbaceniae, and two phytopathogenic isolates were included as outgroups. Qualitative and quantitative morphological characteristics were analyzed using multivariate statistics and were able to distinguish Ceratobasidium, Tulasnella and Thanatephorus genera and separate the isolates of Tulasnella spp. into two groups. Analysis of RAPD (Random Amplified Polymorphic DNA) and ITS rDNA sequences validated the morphological data. Symbionts of O. barbaceniae presented identity to ITS sequences of Ceratobasidium genus, while E. secundum isolates presented identity to two species of Tulasnella. We observed homogeneity among Tulasnella spp. obtained from a single population and from neighboring populations, but there was higher variability among isolates obtained from populations of regions that were farther apart. Morphological data associated with multivariate statistics proved to be a useful tool in the multi-level taxonomy of these orchid-associated fungi and in estimating the diversity of orchid mycorrhizal fungi.Item Effect of cassava mill effluent on biological activity of soil microbial community(Environmental Monitoring and Assessment, 2015-07) Igbinosa, Etinosa O.This study assessed the effect of cassava effluent on soil microbiological characteristics and enzymatic activities were investigated in soil samples. Soil properties and heavy metal concentrations were evaluated using standard soil analytical and spectroscopic methods, respectively. The microbiological parameters measured include microbial biomass carbon, basal soil respiration, catalase, urease, dehydrogenase activities and number of culturable aerobic bacteria, fungi and actinomycetes. The pH and temperature regime vary significantly (p < 0.05) throughout the study period. All other physicochemical parameters studied were significantly different (p < 0.05) higher than the control site. Soil organic carbon content gave significant positive correlations with microbial biomass carbon, basal soil respiration, catalase activity and dehydrogenase activity (r = 0.450, 0.461, 0.574 and 0.591 at p < 0.01), respectively. The quantitative analysis of soil microbial density demonstrates a marked decrease in total culturable numbers of the different microbial groups of the polluted soil samples. Soil contamination decreased catalase, urease and dehydrogenase activities. The findings revealed that soil enzymes can be used as indices of soil contamination and bio-indicator of soil quality.Item Mechanisms of phosphate solubilization by fungal isolates when exposed to different P sources(Annals of Microbiology, 2014-03) Mendes, Gilberto de Oliveira; Freitas, André Luiz Moreira de; Pereira, Olinto Liparini; Silva, Ivo Ribeiro da; Vassilev, Nikolay Bojkov; Costa, Maurício DutraThe use of phosphate-solubilizing fungi is a promising biotechnological strategy in the management of phosphorus (P) fertilization, as it enables the utilization of rock phosphates (RP) or the recovery of P fixed in soil particles. The objective of our study was to evaluate fungal isolates for mechanisms of solubilization of P-bearing compounds, such as AlPO4, FePO4, Ca3(PO4)2, Araxá RP, and Catalão RP. Four fungal isolates obtained from Brazilian soils were characterized in liquid media: Aspergillus niger FS1, Penicillium canescens FS23, Eupenicillium ludwigii FS27, and Penicillium islandicum FS30. A. niger FS1 was the only isolate able to solubilize all of the P sources, solubilizing 71, 36, 100, and 14 % of the P from AlPO4, FePO4, Ca3(PO4)2, and RPs, respectively. Medium acidification was an effective solubilization mechanism, particularly for Ca3(PO4)2. The other P sources were mainly solubilized through organic acids produced by the fungi. Oxalic acid, produced exclusively by A. niger FS1, and citric acid were decisive factors in the solubilization of AlPO4 and FePO4. Penicillium isolates produced more gluconic acid than A. niger FS1 in all treatments. However, this higher production did not result in higher solubilization for any of the P sources, showing that gluconic acid contributes little to the solubilization of the P sources evaluated. The higher capacity of medium acidification and the production of organic acids with stronger metal-complexation activity are characteristics that confer to A. niger FS1 a wider action on insoluble P sources. Consequently, this isolate qualifies as a promising candidate for application in the management of P fertilization.Item Prokaryotic diversity in the rhizosphere of organic, intensive, and transitional Coffee farms in Brazil(Plos One, 2015) Silva, Lívia Carneiro Fidéles; Caldwell, Adam Collins; Silva, Cynthia Canêdo da; Ouverney, Cleber CostaDespite a continuous rise in consumption of coffee over the past 60 years and recent studies showing positive benefits linked to human health, intensive coffee farming practices have been associated with environmental damage, risks to human health, and reductions in biodiversity. In contrast, organic farming has become an increasingly popular alternative, with both environmental and health benefits. This study aimed to characterize and determine the differences in the prokaryotic soil microbiology of three Brazilian coffee farms: one practicing intensive farming, one practicing organic farming, and one undergoing a transition from intensive to organic practices. Soil samples were collected from 20 coffee plant rhizospheres (soil directly influenced by the plant root exudates) and 10 control sites (soil 5 zaway from the coffee plantation) at each of the three farms for a total of 90 samples. Profiling of 16S rRNA gene V4 regions revealed high levels of prokaryotic diversity in all three farms, with thousands of species level operational taxonomic units identified in each farm. Additionally, a statistically significant difference was found between each farm’s coffee rhizosphere microbiome, as well as between coffee rhizosphere soils and control soils. Two groups of prokaryotes associated with the nitrogen cycle, the archaeal genus Candidatus Nitrososphaera and the bacterial order Rhizobiales were found to be abundant and statistically different in composition between the three farms and in inverse relationship to each other. Many of the nitrogen-fixing genera known to enhance plant growth were found in low numbers (e.g. Rhizobium, Agrobacter, Acetobacter, Rhodospirillum, Azospirillum), but the families in which they belong had some of the highest relative abundance in the dataset, suggesting many new groups may exist in these samples that can be further studied as potential plant growth-promoting bacteria to improve coffee production while diminishing negative environmental impacts.Item Biosurfactant, solvents and polymer production by Bacillus subtilis RI4914 and their application for enhanced oil recovery(Fuel, 2016-09-15) Fernandes, P. L.; Rodrigues, E. M.; Paiva, F. R.; Ayupe, B. A. L.; McInerney, M. J.; Tótola, M. R.Effects of nutritional and physicochemical parameters on the production of biosurfactants by Bacillus subtilis RI4914 were evaluated. Effects of three different concentrations of NH4NO3 and/or yeast extract were tested. The treatment with 2.0 g l−1 of NH4NO3 without yeast extract resulted in the highest biosurfactant, organic acid, solvent and polymer concentrations, and this cultivation condition was selected for the residual oil recovery experiments. The lower interfacial tension between crude oil and the culture supernatant was 0.07 mN/m and this result was achieved at NaCl concentrations up to 120 g l−1. Culture fluid with 600 mg l−1 biosurfactant and the polymer produced by the bacterium resulted in residual oil recoveries up to 88%. The attainment of low interfacial tensions and high oil recoveries with the biosurfactant concentrations used at high salinity and temperature indicated that B. subtilis RI4914 is an excellent candidate for application in MEOR.Item Signals of aging associated with lower growth rates in Kluyveromyces lactis cultures under nitrogen limitation(Canadian Journal of Microbiology, 2014) Corrêa, Lygia Fátima da Mata; Passos, Frederico José Vieira; Viloria, Marlene Isabel Vargas; Martins Filho, Olindo Assis; Carvalho, Andréa Teixeira de; Passos, Flávia Maria LopesThe effects of aging on the specific growth rate of Kluyveromyces lactis cultures, as a function of (NH4)2SO4 concentration, were evaluated. The growth kinetic parameters maximum specific growth rate and saturation constant for (NH4)2SO4 were calculated to be 0.44 h−1 and 0.15 mmol·L−1, respectively. Batch cultures were allowed to age for 16 days without influence of cell density or starvation. The specific growth rates of these cultures were determined each day and decreased as the population aged at different nitrogen concentrations. Aging signals (N-acetylglucosamine content of the cell wall, cell dimensions, and apoptosis markers) were measured. Apoptosis markers were detected after 5 days at limiting (NH4)2SO4 concentrations (0.57, 3.80, and 7.60 mmol·L−1) but only after 8 days at a nonlimiting (NH4)2SO4 concentration (38.0 mmol·L−1). Similarly, continuous cultures of K. lactis performed under nitrogen limitation and, at lower dilution rates, accumulated cells exhibiting aging signals. The results demonstrate that aging affects growth rate and raise the question of whether nitrogen limitation accelerates aging. Because aging is correlated with growth rate, and each dilution rate of the continuous cultures tends to select and accumulate cells with a respective age, cultures growing at lower growth rates can be useful to investigate yeast physiological responses, including aging.Item Analysis of Tc1-Mariner elements in Sclerotinia sclerotiorum suggests recent activity and flexible transposases(BMC Microbiology, 2014) Santana, Mateus F.; Silva, José C. F.; Mizubuti, Eduardo S. G.; Araújo, Elza F.; Queiroz, Marisa V.Sclerotinia sclerotiorum is a necrotrophic fungus that is pathogenic to many plants. Genomic analysis of its revealed transposable element expansion that has strongly influenced the evolutionary trajectory of several species. Transposons from the Tc1-Mariner superfamily are thought to be ubiquitous components of fungal genomes and are generally found in low copy numbers with large numbers of deleterious mutations in their transposase coding sequence.This study shows that the genome of S. sclerotiorum has a large number of copies of Tc1-Mariner transposons, and in silico analysis shows evidence that they were recently active. This finding was confirmed by expressed sequence tag (EST) analysis. Fourteen new Tc1-Mariner transposon families that were distributed throughout the genome were identified, and in some cases, due to the excision/retention of introns, different transcripts were observed for the same family, which might be the result of an efficient strategy to circumvent mutations that generate premature stop codons in the RNA sequence. In addition, the presence of these introns shows that the transposase protein has a flexible coding sequence and, consequently, conformation. No evidence for RIP-like gene silencing mechanisms, which are commonly found in fungi, was found in the identified Tc1-Mariner elements, and analysis of the genomic insertion sites of these elements showed that they were widely distributed throughout the genome with some copies located near the 3′ regions of genes. In particular, EST analysis demonstrated that one of these copies was co-expressed with a gene, which showed the potential for these elements to undergo exaptation.Fourteen novel Tc1-Mariner families were characterized. Some families had evidence of introns, which might or might not be excised depending on the family or element in question, and this finding demonstrates a possible strategy for overcoming possible mutations that generate premature stop codons in a RNA sequence. Tc1-Mariner elements likely play an important role in the structure and evolution of the S. sclerotiorum genome.Item Relative importance of soil physico-chemical characteristics and plant species identity to the determination of soil microbial community structure(Applied Soil Ecology, 2015-07) Bonduki, Victor Hugo A.; Melo, Christiane Augusta D.; Tótola, Marcos R.; Ferreira, Francisco A.; Costa, Maurício Dutra; Massenssini, André M.The structure of soil microbial communities is affected by biotic and abiotic environmental factors, such as plant community composition and soil chemical characteristics, among others. However, little is known about the relative importance of these factors on soil microbial community structure. The objective of this study was to verify which factor, soil chemical characteristics or plant species identity, is more important to the determination of soil microbial community structure. For this, a factorial experiment with four soil chemical conditions and five plant species were set in a greenhouse. After 80 days of cultivation, the rhizospheric soil microbial community structure was accessed by a multiplex T-RFLP, and the mycorrhizal colonization of roots and plant shoot dry mass were estimated. Plant species showed similar growth responses to different soil chemical conditions, but exhibited different patterns in the control of root mycorrhizal colonization. A principal component analysis (PCA) was performed using the T-RFLP data set and showed that soil chemical condition is the main factor defining the structure of soil microbial community. Archaeal and bacterial communities showed to be more sensitive to changes in the soil chemical environment, suggesting a greater importance of these microbial groups in plant adaptation.Item Acid and low temperature treatments on Salmonella Enteritidis inoculated in pork and its subsequent survival in simulated gastric fluid(Ciência Rural, 2016-03) Santos, Míriam Teresinha dos; Paes, Simone Albino; Vanetti, Maria Cristina Dantas; Silva, Simone QuintaoThe objective of this study was to evaluate the acid resistance of Salmonella enterica serovar Enteritidis (S. Enteritidis) in stored pork and in simulated gastric fluid (SGF). A culture of S. Enteritidis was subjected to acid treatment prior to inoculation into pork, stored under refrigeration at frozen temperatures and exposed to SGF. The S. Enteritidis CCS3 and ATCC 13076 strains previously subjected to acid treatment (at pH 4.0-5.0) were inoculated in pork and stored at 4[degrees]C and -18[degrees]C. Storage at 40C did not affect the populations of both S. Enteritidis strains. After 84 days at -18[degrees]C, the mean population of both CCS3 and ATCC strains were reduced by 0.8 and 1.5 log cycles, respectively. Prior acid treatment did not enhance the survival of both strains at low temperatures. After acid treatment and low temperature storage, S. Enteritidis ATCC 13076 lost culturability after being exposed to SGF for 10 minutes. In contrast, S. Enteritidis CCS3 was tolerant until three hours of SGF exposure. S. Enteritidis CCS3 submitted to pH 4.0 was more tolerant to SGF exposure than when submitted to pH 4.5, 5.0 and without acid treatment. Therefore, this study indicates that exposure to an acidic and cold environment during processing enhanced the ability of S. Enteritidis to survive in the gastric environment of the human stomach, possibly increasing the risk of a Salmonella infection after consumption of pork.