Microbiologia
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Item Abiotic and biotic degradation of oxo-biodegradable plastic bags by Pleurotus ostreatus(PLoS One, 2014-11-24) Luz, José Maria Rodrigues da; Paes, Sirlaine Albino; Bazzolli, Denise Mara Soares; Tótola, Marcos Rogério; Demuner, Antônio Jacinto; Kasuya, Maria Catarina MegumiIn this study, we evaluated the growth of Pleurotus ostreatus PLO6 using oxobiodegradable plastics as a carbon and energy source. Oxo-biodegradable polymers contain pro-oxidants that accelerate their physical and biological degradation. These polymers were developed to decrease the accumulation of plastic waste in landfills. To study the degradation of the plastic polymers, oxo- biodegradable plastic bags were exposed to sunlight for up to 120 days, and fragments of these bags were used as substrates for P. ostreatus. We observed that physical treatment alone was not sufficient to initiate degradation. Instead, mechanical modifications and reduced titanium oxide (TiO 2 ) concentrations caused by sunlight exposure triggered microbial degradation. The low specificity of lignocellulolytic enzymes and presence of endomycotic nitrogen-fixing microorganisms were also contributing factors in this process.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.Item Action of two herbicides on the microbial activity of soil cultivated with common bean (Phaseolus vulgaris) in conventional-till and no-till systems(Weed Research, 2006-07-06) Santos, J. B.; Jakelaitis, A.; Silva, A. A.; Costa, M. D.; Manabe, A.; Silva, M. C. S.The effects of application of the herbicides fluazifop-p-butyl and fomesafen and the commercial mixture of these herbicides on the microbial activity of a soil, cultivated with common bean under no-till (NTS) and conventional-till (CTS) systems, were evaluated. Microbial respiration was monitored for 63 days after application (DAA) of the herbicides, and the following evaluated at 12 and 51 DAA: microbial biomass carbon (MBC), microbial quotient (qMIC), metabolic quotient (qCO2), percentage of bean root colonisation by mycorrhizal fungi and grain yield at the end of the cycle. A greater microbial respiratory rate was observed under NTS, with fluazifop-p-butyl providing the lowest respiration. At 12 DAA, MBC and qMIC were most affected negatively by fomesafen and by the commercial mixture of the two herbicides. Mycorrhizal colonisation was affected by the herbicides only at 12 DAA under CTS; however, in both periods, the highest value was found under NTS. All the herbicides caused a decrease in the MBC and qMIC values at 51 DAA; the qCO2, which is related to the soil system stability, indicated a greater NTS balance over CTS. The herbicide fomesafen induced lower stability in the system. Lower grain yield was obtained without weed control (no herbicides) and with fomesafen-only treatments, which may be attributed to the high weed infestation in the experimental area.Item The activity of b-galactosidase and lactose metabolism in Kluyveromyces lactis cultured in cheese whey as a function of growth rate(Journal of Applied Microbiology, 2007-09-13) Ornelas, A.P.; Silveira, W.B.; Sampaio, F.C.; Passos, F.M.L.Kluyveromyces lactis was cultured in cheese whey permeate on both batch and continuous mode to investigate the effect of time course and growth rate on beta-galactosidase activity, lactose consumption, ethanol production and protein profiles of the cells. Cheese whey was the substrate to grow K. lactis as a batch or continuous culture. In order to precise the specific growth rate for maximum beta-galactosidase activity a continuous culture was performed at five dilution (growth) rates ranging from 0.06, 0.09, 0.12, 0.18 to 0.24 h(-1). The kinetics of lactose consumption and ethanol production were also evaluated. On both batch and continuous culture a respirofermentative metabolism was detected. The growth stage for maximum beta-gal activity was found to be at the transition between late exponential and entrance of stationary growth phase of batch cultures. Fractionating that transition stage in several growth rates at continuous culture a maximum beta-galactosidase activity at 0.24 h(-1) was observed. Following that stage beta-gal activity undergoes a decline which does not correlate to the density of its corresponding protein band on the gel prepared from the same samples. The maximum beta-galactosidase activity per unit of cell mass was found to be 341.18 mmol ONP min(-1) g(-1) at a dilution rate of 0.24 h(-1). The physiology of K. lactis growing in cheese whey permeate can proven useful to optimize the conversion of that substrate in biomass rich in beta-gal or in ethanol fuel. In addition to increasing the native enzyme the conditions established here can be set to increase yields of recombinant protein production based on the LAC4 promoter in K. lactis host.Item The activity of β-galactosidase and lactose metabolism in Kluyveromyces lactis cultured in cheese whey as a function of growth rate(Journal of Applied Microbiology, 2007-11-27) Ornelas, A. P.; Silveira, W. B.; Sampaio, F. C.; Passos, F. M. LKluyveromyces lactis was cultured in cheese whey permeate on both batch and continuous mode to investigate the effect of time course and growth rate on β-galactosidase activity, lactose consumption, ethanol production and protein profiles of the cells. Cheese whey was the substrate to grow K. lactis as a batch or continuous culture. In order to precise the specific growth rate for maximum β‐galactosidase activity a continuous culture was performed at five dilution (growth) rates ranging from 0·06, 0·09, 0·12, 0·18 to 0·24 h−1. The kinetics of lactose consumption and ethanol production were also evaluated. On both batch and continuous culture a respirofermentative metabolism was detected. The growth stage for maximum β‐gal activity was found to be at the transition between late exponential and entrance of stationary growth phase of batch cultures. Fractionating that transition stage in several growth rates at continuous culture a maximum β‐galactosidase activity at 0·24 h−1 was observed. Following that stage β‐gal activity undergoes a decline which does not correlate to the density of its corresponding protein band on the gel prepared from the same samples.The maximum β‐galactosidase activity per unit of cell mass was found to be 341·18 mmol ONP min−1 g−1 at a dilution rate of 0·24 h−1.The physiology of K. lactis growing in cheese whey permeate can proven useful to optimize the conversion of that substrate in biomass rich in β‐gal or in ethanol fuel. In addition to increasing the native enzyme the conditions established here can be set to increase yields of recombinant protein production based on the LAC4 promoter in K. lactis host.Item Acúmulo de ácido oxálico e cristais de cálcio em ectomicorrizas de eucalipto. I- produção de ácido oxálico e concentração de nutrientes em raízes laterais finas colonizadas por fungos ectomicorrízicos(Revista Brasileira de Ciência do Solo, 2009-03) Gonzalez, Jhon Alexander Zambrano; Costa, Maurício Dutra; Silva, Ivo Ribeiro; Neves, Júlio César Lima; Barros, Nairam Félix de; Borges, Arnaldo ChaerO eucalipto apresenta ótimo crescimento em solo com baixa fertilidade, mas pouco se sabe sobre a participação das ectomicorrizas e de ácidos orgânicos na aquisição e no acúmulo de nutrientes pela planta em campo. A produção de ácido oxálico e sua relação com as concentrações de P, Ca, Mg e K foram avaliadas em ectomicorrizas e raízes laterais finas de híbrido de Eucalyptus grandis x Eucalyptus urophylla, de 2,5 anos de idade, na região de Viçosa, MG. A área de estudo apresenta topografia típica em meia laranja, de vertente côncavo-convexa. Foram também avaliadas as concentrações desse composto no solo rizosférico, não rizosférico e ectomicorrizosférico. As maiores percentagens de colonização micorrízica foram observadas na área de encosta, onde havia limitada disponibilidade de nutrientes e alta saturação de Al. As concentrações de ácido oxálico + oxalato corresponderam, em mg kg-1: folhas, 324,6; ectomicorrizas, 208,3; raízes laterais finas não colonizadas, 183,1. Já no solo, as concentrações foram maiores no solo ectomicorrizosférico, com 183,7 mg kg-1, seguido pelo solo rizosférico, com 134,3 mg kg-1, e pelo solo não rizosférico, com 76,0 mg kg-1. As maiores concentrações de ácido oxálico e P (p < 0,05) nas ectomicorrizas da área do topo, 117,3 mg kg-1 e 6,3 g kg-1, respectivamente, sugerem que as populações de fungos ectomicorrízicos nesta área têm papel importante na solubilização e disponibilização de nutrientes para o hospedeiro. Não foram observadas correlações positivas significativas entre a produção de ácido oxálico e as concentrações de Ca nas raízes laterais finas e nas ectomicorrizas de eucalipto.Item Acúmulo de ácido oxálico e cristais de cálcio em ectomicorrizas de eucalipto. II- formação de cristais de oxalato de cálcio induzida por fungos ectomicorrízicos em raízes laterais finas(Revista Brasileira de Ciência do Solo, 2009-03) Gonzalez, Jhon Alexander Zambrano; Costa, Maurício Dutra; Silva, Ivo Ribeiro; Neves, Júlio César Lima; Barros, Nairam Félix de; Borges, Arnaldo ChaerO eucalipto é eficiente na aquisição de Ca do solo, mas pouco se sabe sobre a participação das ectomicorrizas e dos ácidos orgânicos nesse processo em campo. O acúmulo de cristais de Ca (CaOx) foi avaliado em, aproximadamente, 2.100 raízes laterais finas e ectomicorrizas do híbrido de Eucalyptus grandis x Eucalyptus urophylla, cultivado por 2,5 anos em área com topografia típica em meia laranja, com vertente côncavo-convexa, na região de Viçosa, MG. Técnicas de microscopia óptica e microscopia eletrônica de varredura foram usadas para a visualização dos CaOx. Em 73,7 % das raízes, ocorreu abundante acúmulo de drusas e grânulos de CaOx nas células do córtex. A presença conspícua de CaOx foi observada em 56,2 % das ectomicorrizas e em 17,5 % das raízes laterais finas não colonizadas, evidenciando o papel das micorrizas no acúmulo de Ca em eucalipto. A forma predominante dos CaOx foram as drusas nas ectomicorrizas e os grânulos cristalinos nas raízes. Os dez morfotipos de ectomicorrizas observados na área diferiram quanto à presença e à morfologia dos CaOx, o que pode representar distintas capacidades dos fungos ectomicorrízicos em fornecer Ca para a planta hospedeira. A análise da superfície do manto das ectomicorrizas por microscopia eletrônica de varredura não evidenciou a presença de CaOx nessa estrutura, confirmando que, nas condições avaliadas, o acúmulo de cristais limita-se ao córtex radicular. Este é o primeiro relato da ocorrência de CaOx em ectomicorrizas de eucalipto no Brasil, com dados que comprovam que há mecanismos de armazenamento de Ca nas ectomicorrizas em áreas com baixa disponibilidade do elemento.Item Acyl homoserine lactone changes the abundance of proteins and the levels of organic acids associated with stationary phase in Salmonella Enteritidis(Microbial Pathogenesis, 2016-12-01) Almeida, Felipe Alvesde; Pimentel-Filho, Natan de Jesus; Carrijo, Lanna Clícia; Bento, Claudia Braga Pereira; Baracat-Pereira, Maria Cristina; Pinto, Uelinton Manoel; Oliveira, Leandro Licursi de; Vanetti, Maria Cristina DantasQuorum sensing (QS) is cell-cell communication mechanism mediated by signaling molecules known as autoinducers (AIs) that lead to differential gene expression. Salmonella is unable to synthesize the AI-1 acyl homoserine lactone (AHL), but is able to recognize AHLs produced by other microorganisms through SdiA protein. Our study aimed to evaluate the influence of AI-1 on the abundance of proteins and the levels of organic acids of Salmonella Enteritidis. The presence of N-dodecyl-homoserine lactone (C12-HSL) did not interfere on the growth or the total amount of extracted proteins of Salmonella. However, the abundance of the proteins PheT, HtpG, PtsI, Adi, TalB, PmgI (or GpmI), Eno, and PykF enhanced while the abundance of the proteins RplB, RplE, RpsB, Tsf, OmpA, OmpC, OmpD, and GapA decreased when Salmonella Enteritidis was anaerobically cultivated in the presence of C12-HSL. Additionally, the bacterium produced less succinic, lactic, and acetic acids in the presence of C12-HSL. However, the concentration of extracellular formic acid reached 20.46 mM after 24 h and was not detected when the growth was in the absence of AI-1. Considering the cultivation period for protein extraction, their abundance, process and function, as well as the levels of organic acids, we observed in cells cultivated in presence of C12-HSL a correlation with what is described in the literature as entry into the stationary phase of growth, mainly related to nitrogen and amino acid starvation and acid stress. Further studies are needed in order to determine the specific role of the differentially abundant proteins and extracellular organic acids secreted by Salmonella in the presence of quorum sensing signaling molecules.Item Acyl homoserine lactone-based quorum sensing stimulates biofilm formation by Salmonella Enteritidis in anaerobic conditions(Archives of Microbiology, 2016-11-12) Almeida, Felipe Alves de; Pimentel-Filho, Natan de Jesus; Pinto, Uelinton Manoel; Mantovani, Hilário Cuquetto; Oliveira, Leandro Licursi de; Vanetti, Maria Cristina DantasQuorum sensing regulates a variety of phenotypes in bacteria including the production of virulence factors. Salmonella spp. have quorum sensing systems mediated by three autoinducers (AI-1, AI-2, and AI-3). The AI-1-mediated system is incomplete in that the bacterium relies on the synthesis of signaling molecules by other microorganisms. This study aimed to evaluate the influence of the AI-1 N-dodecanoyl-DL-homoserine lactone (C12-HSL) on the growth, motility, adhesion, and biofilm formation of Salmonella enterica serovar Enteritidis PT4 578 on a polystyrene surface. Experiments were conducted at 37 °C in anaerobic tryptone soy broth supplemented with C12-HSL and/or a mixture of four synthetic furanones, at the concentration of 50 nM each. The planktonic growth, adhesion, swarming, and twitching motility were not altered in the presence of C12-HSL and/or furanones under anaerobic conditions. However, C12-HSL induced biofilm formation after 36 h of cultivation as determined by quantification of biofilm formation, by enumeration of adhered cells to polystyrene coupons, and finally by imaging the presence of multilayered cells on an epifluorescence microscope. When furanones were present in the medium, an antagonistic effect against C12-HSL on the biofilm development was observed. The results demonstrate an induction of biofilm formation in Salmonella Enteritidis by AI-1 under anaerobic conditions. Considering that Salmonella does not produce AI-1 but respond to it, C12-HSL synthesized by other bacterial species could trigger biofilm formation by this pathogen in conditions that are relevant for its pathogenesis.Item Addressing global ruminant agricultural challenges through understanding the rumen microbiome: past, present, and future(Frontiers in Microbiology, 2019) Mantovani, Hilario C.; Huws, Sharon A.; Creevey, Christopher J.; Oyama, Linda B.; Mizrahi, Itzhak; Denman, Stuart E.; Popova, Milka; Muñoz-Tamayo, Rafael; Forano, Evelyne; Waters, Sinead M.; Hess, Matthias; Tapio, Ilma; Smidt, Hauke; Krizsan, Sophie J.; Yáñez-Ruiz, David R.; Belanche, Alejandro; Guan, Leluo; et alThe rumen is a complex ecosystem composed of anaerobic bacteria, protozoa, fungi, methanogenic archaea and phages. These microbes interact closely to breakdown plant material that cannot be digested by humans, whilst providing metabolic energy to the host and, in the case of archaea, producing methane. Consequently, ruminants produce meat and milk, which are rich in high-quality protein, vitamins and minerals, and therefore contribute to food security. As the world population is predicted to reach approximately 9.7 billion by 2050, an increase in ruminant production to satisfy global protein demand is necessary, despite limited land availability, and whilst ensuring environmental impact is minimized. Although challenging, these goals can be met, but depend on our understanding of the rumen microbiome. Attempts to manipulate the rumen microbiome to benefit global agricultural challenges have been ongoing for decades with limited success, mostly due to the lack of a detailed understanding of this microbiome and our limited ability to culture most of these microbes outside the rumen. The potential to manipulate the rumen microbiome and meet global livestock challenges through animal breeding and introduction of dietary interventions during early life have recently emerged as promising new technologies. Our inability to phenotype ruminants in a high-throughput manner has also hampered progress, although the recent increase in “omic” data may allow further development of mathematical models and rumen microbial gene biomarkers as proxies. Advances in computational tools, high-throughput sequencing technologies and cultivation-independent “omics” approaches continue to revolutionize our understanding of the rumen microbiome. This will ultimately provide the knowledge framework needed to solve current and future ruminant livestock challenges.Item Agroecological coffee management increases arbuscular mycorrhizal fungi diversity(Plos One, 2019) Prates Júnior, Paulo; Moreira, Bruno Coutinho; Silva, Marliane de Cássia Soares da; Veloso, Tomas Gomes Reis; Stürmer, Sidney Luiz; Fernandes, Raphael Bragança Alves; Mendonça, Eduardo de Sá; Kasuya, Maria Catarina MegumiAgroecology aims to maintain ecosystem services by minimizing the impact of agriculture and promoting the use of biological potential. Arbuscular mycorrhizal fungi (AMF) are elements which are key to improving crop productivity and soil quality. It is pertinent to understand how agricultural management in the tropics affects the AMF spatio-temporal community composition, especially in crops of global importance, such as coffee (Coffea arabica L.). Soil and root samples were collected from three localities under three management systems (agroecological, conventional and forest fragment), during the phenological stages of coffee (flowering, grain filling, harvesting). Spores were extracted for morphological identification and molecular community analysis by PCR–DGGE. Dendrograms were prepared and the bands were sequenced and analyzed by bioinformatics. No differences were observed in the richness of morphospecies between management systems, localities and period, but little is known about tropical species. Molecular analysis showed that the agroecological management system was similar to natural forest and with a higher diversity indices than conventional management. Locality and period of sample affect AMF community composition. It is necessary to associate classical taxonomic evaluations with molecular biological techniques because different approaches can lead to different outcomes. This study contributes to the understanding of the impact of agriculture management systems on AMF and provides evidence that agroecology is a management system applicable to sustainable coffee production.Item Aliphatic hydrocarbon enhances phenanthrene degradation by autochthonous prokaryotic communities from a pristine seawater(Microbial Ecology, 2017-10-03) Rodrigues, Edmo Montes; Morais, Daniel Kumazawa; Pylro, Victor Satler; Redmile-Gordon, Marc; Oliveira, uraci Alves de; Roesch, Luiz Fernando Wurdig; Cesar, Dionéia Evangelista; Tótola, Marcos RogérioThe microbial diversity and functioning around oceanic islands is poorly described, despite its importance for ecosystem homeostasis. Here, we aimed to verify the occurrence of microbe-driven phenanthrene co-oxidation in the seawater surrounding the Trindade Island (Brazil). We also used Next-Generation Sequencing to evaluate the effects of aliphatic and polycyclic aromatic hydrocarbons (PAHs) on these microbial community assemblies. Microcosms containing seawater from the island enriched with either labelled (9^-14C) or non-labelled phenanthrene together with hexadecane, weathered oil, fluoranthene or pyrene, and combinations of these compounds were incubated. Biodegradation of phenanthrene-9^-14C was negatively affected in the presence of weathered oil and PAHs but increased in the presence of hexadecane. PAH contamination caused shifts in the seawater microbial community—from a highly diverse one dominated by Alphaproteobacteria to less diverse communities dominated by Gammaproteobacteria. Furthermore, the combination of PAHs exerted a compounded negative influence on the microbial community, reducing its diversity and thus functional capacity of the ecosystem. These results advance our understanding of bacterial community dynamics in response to contrasting qualities of hydrocarbon contamination. This understanding is fundamental in the application and monitoring of bioremediation strategies if accidents involving oil spillages occur near Trindade Island and similar ecosystems.Item Análise comparativa do genoma mitocondrial do fungo Colletotrichum lindemuthianum, agente causal da antracnose em feijão comum(Applied Microbiology and Biotechnology, 2018-02-16) Queiroz, Casley Borges de; Santana, Mateus Ferreira; Vidigal, Pedro M. Pereira; Queiroz, Marisa Vieira deFungi of the genus Colletotrichum are economically important and are used as models in plant-pathogen interaction studies. In this study, the complete mitochondrial genomes of two Colletotrichum lindemuthianum isolates were sequenced and compared with the mitochondrial genomes of seven species of Colletotrichum. The mitochondrial genome of C. lindemuthianum is a typical circular molecule 37,446 bp (isolate 89 A2 2-3) and 37,440 bp (isolate 83.501) in length. The difference of six nucleotides between the two genomes is the result of a deletion in the ribosomal protein S3 (rps3) gene in the 83.501 isolate. In addition, substitution of adenine for guanine within the rps3 gene in the mitochondrial genome of the 83.501 isolate was observed. Compared to the previously sequenced C. lindemuthianum mitochondrial genome, an exon no annotated in the cytochrome c oxidase I (cox1) gene and a non-conserved open reading frame (ncORF) were observed. The size of the mitochondrial genomes of the seven species of Colletotrichum was highly variable, being attributed mainly to the ncORF, ranging from one to 10 and also from introns ranging from one to 11 and which encode a total of up to nine homing endonucleases. This paper reports for the first time by means of transcriptome that then ncORFs are transcribed in Colletotrichum spp. Phylogeny data revealed that core mitochondrial genes could be used as an alternative in phylogenetic relationship studies in Colletotrichum spp. This work contributes to the genetic and biological knowledge of Colletotrichum spp., which is of great economic and scientific importance.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 Antimicrobial activity and mineral composition of shiitake mushrooms cultivated on agricultural waste(Brazilian Archives of Biology and Technology, 2011-09) Kasuya, Maria Catarina Megumi; Casaril, Kérley Braga Pereira Bento; Vanetti, Maria Cristina DantasThe antimicrobial activity and mineral composition of shiitake mushrooms were evaluated in four isolates of Lentinula edodes. Mushrooms were cultivated on artificial logs, based on eucalyptus sawdust enriched with 20% rice, wheat, or soybean bran, or combination of 10% of two of these supplements. The substrates were humidified with a 0.1% mate tea extract or water. Logs of Eucalyptus grandis were also used to cultivate the shiitake mushrooms. The antimicrobial activity of an aqueous extract, corresponding to 40 mg of mushroom dry matter, was in some cases, depending on the isolate, able to inhibit both Bacillus subtilis and Escherichia coli K-12, independent of substrate composition or the growth stage of the mushrooms. Nitrogen, phosphorus, potassium, magnesium and calcium concentrations varied according to the substrate on which the mushrooms were cultivated, being, generally, higher with cultivation on artificial rather than natural eucalyptus logs. It could be concluded that, in addition to the fungal isolate, substrate composition and, processing methods must be considered during the production of antimicrobial substance(s) as well as in the mushroom nutritional compositionItem Antimicrobial profile screening of two oils of Copaifera genus(Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 2011-12-01) Pieri, F.A.; Silva, V.O.; Souza, C.F.; Costa, J.C.M.; Santos, L.F.; Moreira, M.A.S.O objetivo deste estudo foi identificar a atividade inibitória de óleos de copaíba sobre o crescimento dos micro-organismos: Shigella flexneri, Klebsiella pneumoniae, Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Citrobacter freundi, Staphylococcus aureus, Actinobacillus pleuropneumoniae e Haemophilus parasuis. Foi realizado um teste de difusão em ágar com duas soluções a 10% de óleos de copaíba obtidos de duas diferentes espécies de copaíba (Copaifera officinalis e C. langsdorffii) e um controle negativo com tween 80 e água. Os isolados clínicos de H. parasuis e A. pleuropneumoniae foram incubados em microaerofilia. Os resultados mostraram três espécies Gram-negativas inibidas por ambas as soluções de óleo de copaíba: E. coli, P. aeruginosa e S. flexneri. Na inibição de P. aeruginosa o óleo de C. officinalis foi superior ao de C. langsdorffii (P<0,05). Todas as cepas de S. aureus tiveram seu crescimento inibido pelas soluções no ensaio, sem diferença estatística entre os halos. Estes resultados sugerem que o óleo de copaíba pode ser uma fonte potencial de compostos inibitórios para ser utilizada como antimicrobianos no tratamento de infecções humanas e animais e conservação de alimentos.Item Antimicrobial resistance, biofilm formation and virulence reveal Actinobacillus pleuropneumoniae strains' pathogenicity complexity(Research in Veterinary Science, 2018-06) Pereira, Monalessa Fábia; Rossi, Ciro César; Seide, Larissa Eler; Martins Filho, Sebastião; Dolinski, Cláudia de Melo; Bazzolli, Denise Mara SoaresPorcine pleuropneumonia is an important cause of lowered productivity and economic loss in the pig industry worldwide, associated primarily with Actinobacillus pleuropneumoniae infection. Its colonization and persistence within the upper respiratory tract of affected pigs depends upon interactions between a number of genetically controlled virulence factors, such as pore-forming repeats-in-toxin exoproteins, biofilm formation, and antimicrobial resistance. This study investigated correlations between biofilm-forming capacity, antimicrobial resistance, and virulence of A. pleuropneumoniae obtained from clinical outbreaks of disease, using a Galleria mellonella alternative infection model. Results suggest that virulence is diverse amongst the 21 strains of A. pleuropneumoniae examined and biofilm formation correlated with genetic control of antimicrobial resistance.Item Antioxidant activities, total phenolics and metal contents in Pleurotus ostreatus mushrooms enriched with iron, zinc or lithium(LWT - Food Science and Technology, 2013-12) Assunção, Laélia Soares de; Vieira, Patrícia A. Fontes; Gontijo, Douglas C.; Vieira, Breno César; Fontes, Edimar A. F.; Leite, João Paulo V.; Oliveira, Maria Goreti de Almeida; Kasuya, Maria Catarina M.Food enrichment with minerals has been a method of increasing health benefits. Edible Pleurotus ostreatus mushrooms are a source of dietary elements and antioxidants. In this work, the phenolic content, antioxidant activity and metal content of P. ostreatus grown on coffee husk, enriched with iron (Fe), zinc (Zn) and lithium (Li) were investigated. The content of phenolic compounds was found to be different in mushroom enriched with Fe, Zn or Li. In DPPH test, a difference was observed in antioxidant activity; however, no difference in this activity was observed using β-carotene/linoleic acid test. The enrichment with minerals promoted decrease in the content of Fe in three enriched samples. There was no change in the content of Zn in the samples. Observed the accumulation of Li in the sample enriched with LiCl and P. ostreatus enriched with Fe, Zn or Li provided minerals such as K, P, Fe, Zn, Li and Cu. In addition, no heavy metals such as Ni, Cr and Cd were detected, and only low levels of Pb and Al were observed. The study was the first report to demonstrate the understanding of how mineral supplementation reduces antioxidant activity in P. ostreatus enriched with iron, zinc or lithium.Item Antioxidant, antiproliferative, and immunostimulatory effects of cell wall α-d-mannan fractions from Kluyveromyces marxianus(International Journal of Biological Macromolecules, 2017-11-11) Galinari, Éder; Almeida-Lima, Jailma; Macedo, Gorete Ribeiro; Mantovani, Hilário Cuquetto; Rocha, Hugo Alexandre OliveiraThis study evaluated the antioxidant, antiproliferative, and immunostimulatory properties of cell wall α-d-mannan fractions from yeast Kluyveromyces marxianus CCT7735. Filter centrifugation was used to obtain four fractions (KMM-1, KMM-2, KMM-3, and KMM-4) with molecular weight ranging from 7.6 to 75.1 kDa. KMM-1 and KMM-2 comprised D-mannose with traces of D-glucose, whereas other fractions contained only D-mannose. Total sugar found in samples ranged from 85.9% to 96.1%, while protein and phenolic contents were 1.21% and 0.41%, respectively. Although only KMM-1 was able to scavenge superoxide radicals, all fractions presented total antioxidant capacity as well as reducing power, hydroxyl-radical scavenging, and copper- and iron-chelating activities. No fraction was cytotoxic to HeLa cells. However, all samples inhibited the proliferation of the tumor cell Hep-G2 and presented minor cytotoxicity against normal 3T3 cells. All fractions showed mitogenic activity in macrophages and all, except KMM-4, induced nitric oxide production in macrophages, suggestive of their immunostimulatory effects.Item Applying functional metagenomics to search for novel lignocellulosic enzymes in a microbial consortium derived from a thermophilic composting phase of sugarcane bagasse and cow manure(Antonie van Leeuwenhoek, 2016-06-27) Colombo, Lívia Tavares; Oliveira, Marcelo Nagem Valério de; Carneiro, Deisy Guimarães; Souza, Robson Assis de; Alvim, Mariana Caroline Tocantins; Santos, Josenilda Carlos dos; Silva, Cynthia Canêdo da; Vidigal, Pedro Marcus Pereira; Silveira, Wendel Batista da; Passos, Flávia Maria LopesEnvironments where lignocellulosic biomass is naturally decomposed are sources for discovery of new hydrolytic enzymes that can reduce the high cost of enzymatic cocktails for second-generation ethanol production. Metagenomic analysis was applied to discover genes coding carbohydrate-depleting enzymes from a microbial laboratory subculture using a mix of sugarcane bagasse and cow manure in the thermophilic composting phase. From a fosmid library, 182 clones had the ability to hydrolyse carbohydrate. Sequencing of 30 fosmids resulted in 12 contigs encoding 34 putative carbohydrate-active enzymes belonging to 17 glycosyl hydrolase (GH) families. One third of the putative proteins belong to the GH3 family, which includes β-glucosidase enzymes known to be important in the cellulose-deconstruction process but present with low activity in commercial enzyme preparations. Phylogenetic analysis of the amino acid sequences of seven selected proteins, including three β-glucosidases, showed low relatedness with protein sequences deposited in databases. These findings highlight microbial consortia obtained from a mixture of decomposing biomass residues, such as sugar cane bagasse and cow manure, as a rich resource of novel enzymes potentially useful in biotechnology for saccharification of lignocellulosic substrate.