Microbiologia
URI permanente desta comunidadehttps://locus.ufv.br/handle/123456789/11840
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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 Effect of different initial pH on the growth of Streptococcus macedonicus and Streptococcus thermophilus strains(International Dairy Journal, 2018-11) Duarte, Vinícius da Silva; Tarrah, Armin; Noal, Valentina; Giaretta, Sabrina; Treu, Laura; Corich, Viviana; Giacomini, AlessioStreptococcus thermophilus and Streptococcus macedonicus are two known species of the genus related to foods, but little information is available for the latter species, introduced quite recently. Streptococci are widely used as starter cultures to rapidly lower food pH, thus inhibiting the development of deleterious and pathogenic microbes. We compared the growth kinetics over 24 h of nine S. thermophilus and eight S. macedonicus strains, isolated from various dairy environments, at three different pH values, namely 7.0, 6.0 and 5.5. The Gompertz model was used to estimate the three main growth parameters, namely lag phase duration (λ), maximum growth rate (μmax) and maximum cell number at the stationary phase (Nmax). Our results report significantly shorter lag phases and faster growth rates at all the pH conditions tested for S. macedonicus strains, thus indicating this species as a possible interesting substitute for S. thermophilus in selected food productions.Item Genetic variability and chromosome-length polymorphisms of the witches’ broom pathogen Crinipellis perniciosa from various plant hosts in South America(Mycological Research, 2006-07) Queiroz, Marisa V.; Rincones, Johana; Mazotti, Gabriel D.; Griffith, Gareth W.; Pomela, Alan; Figueira, Antonio; Leal Jr., Gildemberg A.; Pereira, Jorge F.; Azevedo, Ricardo A.; Pereira, Gonçalo A. G.; Meinhardt, Lyndel W.Crinipellis perniciosa has been classified into at least four known biotypes associated with members of unrelated plant families. In this study, genetic variability is shown for 27 C (Cacao), 4 S (Solanum), and 7 L biotype (Liana) isolates of C. perniciosa collected from different regions of Brazil and South America. The objective was to investigate the genetic variability of the pathogen in the cacao-producing region of Bahia, Brazil, and elsewhere, through microsatellite analysis, and attempt to identify possible correlations between host specificity and electrophoretic karyotypes. The PCR-banding patterns were found to vary both within and between the different biotypes, and a correlation was established between the PCR-banding patterns and the chromosomal-banding patterns of each isolate. Microsatellite and chromosomal patterns among all of the L and S biotype isolates were distinctly different from the C biotypes analysed. A higher degree of genetic and chromosomal variability was found among C biotype isolates from the Amazon in comparison with C biotype isolates from Bahia, which seems to be comprised of only two main genotypes. This finding has important implications to the current cacao-breeding programme in Brazil.Item Inhibition of Listeria monocytogenes by a lactic acid bacterium isolated from Italian salami(Food Microbiology, 2005-07-18) Carvalho, Ana Andréa Teixeira de; Paula, Rosinéa Aparecida de; Mantovani, Hilário C.; Moraes, Célia Alencar deListeria monocytogenes is an opportunistic psychrotroph foodborne pathogen that has been used as a model organism to study the efficacy of many different preservation methods. This work aimed to test the antilisterial activity of lactic acid bacteria isolated from Italian salami and study the development of resistance. Isolates were obtained from naturally fermented Italian salami and cultures that retained activity in the supernatants after pH neutralization and catalase treatment were further characterized. The isolate showing highest inhibitory activity (PD 6.9) was tested for sensibility to proteases, heat and pH. To evaluate if resistance developed, sensitive strains were transferred with sub-lethal doses of the partially purified inhibitory substance and then inoculated into media containing higher doses of the extract. Isolate PD 6.9 inhibited several L. monocytogenes strains obtained from different origins and retained its activity over a wide range of pH and temperature. When increasing concentrations (10–100 AU ml−1) of the partially purified inhibitory substance were added to culture media, growth of L. monocytogenes did not occur even after 12 h of incubation. Cultures of Listeria that were transferred with sub-lethal doses (10 AU ml−1) of the partially purified inhibitory substance could resist higher doses of the extract (50 AU ml−1), but were inhibited when the concentration was further increased (100 AU ml−1). These results indicate that isolate PD 6.9 could potentially be used as a bioprotective culture for salami fermentation.Item A new biostimulation approach based on the concept of remaining P for soil bioremediation(Journal of Environmental Management, 2017-11-28) Júlio, Aline Daniela Lopes; Fernandes, Rita de Cássia Rocha; Costa, Maurício Dutra; Neves, Júlio César Lima; Rodrigues, Edmo Montes; Tótola, Marcos RogérioC:N:P ratio is generally adopted to estimate the amount of nitrogen and phosphorus to be added to soils to accelerate biodegradation of organic contaminants. However, differences in P fixation among soils lead to varying amounts of available P when a specific dose of the element is applied to different soils. Thus, the application of fertilizers to achieve a previously established C:P ratio leads to biodegradation rates that can be lower than the theoretical maximum. In this study, we developed an equation to estimate the dose of P required to maximize organic contaminant biodegradation in soils as a function of remaining P (P-rem), using diesel as a model contaminant. The soils were contaminated with diesel and received six doses of P. CO 2 emission was used to estimate biodegradation of hydrocarbons. Biodegradation increased with P doses. The P level that provided the highest hydrocarbon biodegradation rate showed linear and negative correlation with P-rem. The result shows that the requirement for P decreases as the P-rem of the soil increases (or the P-fixing capacity decreases). The dose of P recommended to maximize hydrocarbon biodegradation rate in soil can be estimated by the formula P (mg/dm 3 ) 1⁄4 436.5e5.39 Â P-rem (mg/L).Item Physiological implications of trehalose in the ectomycorrhizal fungus Pisolithus sp. under thermal stress(Journal of Thermal Biology, 2007-01) Tótola, Marcos R.; Ferreira, Adão S.; Borges, Arnaldo C.The trehalose cycle in filamentous fungi has been suggested to be an important mechanism of tolerance against adverse stress conditions, particularly in thermal stress. Here, we demonstrate that trehalose and trehalase activity can be involved as an additional mechanism in development of thermotolerance in Pisolithus sp. In response to heat shock at 42 °C, an accumulation of intracellular trehalose and an increase in trehalase activity in the mycelial mass of the fungus was observed, when compared to the control treatment (28 °C). In vitro, assays showed that trehalose had a protective effect on β-glucosidase activity under thermal stress. Therefore, trehalose production may be an important mechanism of protection in ectomycorrhizal fungi, and this capacity could be used in the selection of isolates with greater capacity for adaptation to environmental stress.Item Prospect, isolation, and characterization of microorganisms for potential use in cases of oil bioremediation along the coast of Trindade Island, Brazil(Journal of Environmental Management, 2015-06-01) Rodrigues, Edmo M.; Kalks, Karlos H.M.; Tótola, Marcos R.In the present study, acrylic coupons with a thin layer of oil on the surface were incubated in the coastal water of Trindade Island, Brazil, for 60 days. The microorganisms adhered to the coupons were isolated using enrichment medium with hexadecane and naphthalene as the sole carbon and energy source. A total of 15 bacterial isolates were obtained, and the ability of these isolates to use different hydrocarbons as the source of carbon and energy was investigated. None of the isolates produced biosurfactants under our experimental conditions. Subsequently, identification methods such as partial sequencing of the 16S rRNA gene and analysis of fatty acids (MIDI) profile were employed. Among the 15 isolates, representatives of Actinobacteria, Firmicutes, and Alphaproteobacteria were detected. The isolates Rhodococcus rhodochrous TRN7 and Nocardia farcinica TRH1 were able to use all the hydrocarbons added to the culture medium (toluene, octane, xylene, naphthalene, phenanthrene, pyrene, hexadecane, anthracene, eicosane, tetracosane, triacontane, and pentacontane). Polymerase chain reaction amplification of the DNA isolated by employing primers for catechol 2,3-dioxygenase, alkane dehydrogenase and the alpha subunit of hydroxylating dioxygenases polycyclic aromatic hydrocarbon rings genes demonstrated that various isolates capable of utilizing hydrocarbons do not exhibit genes of known routes of catabolism, suggesting the existence of unknown catabolic pathways in these microorganisms. Our findings suggest that the microbiota associated to the coast of tropical oceanic islands has the ability to assist in environmental regeneration in cases of accidents involving oil spills in its shore. Thus, it motivates studies to map bioremediation strategies using the autochthonous microbiota from these environments.Item Small heat shock proteins in the development of thermotolerance in Pisolithus sp.(Journal of Thermal Biology, 2005-12) Ferreira, Adão S.; Kasuya, Maria C. M.; Araújo, Elza F.; Borges, Arnaldo C.; Tótola, Marco. R.Small heat shock proteins (HSPs) have been shown to confer thermotolerance in many organisms. Here, we demonstrate that small HSPs (sHSPs) can also be involved in development of thermotolerance in Pisolithus sp. In heat shock response, Pisolithus isolate RV82 synthesized proteins of molecular mass 28, 26 and 15–18 kDa. These group of proteins are synthesized when mycelial mass are exposed to heat shock temperature (42 °C) for short period (30 min) and incubated back at 28 °C, the optimal temperature for growth. Our results show sHSPs are an important biochemical alteration in ectomycorrhizal fungi under thermal stress.Item Temperature and pH influence the susceptibility of Salmonella Typhimurium to nisin combined with EDTA(Food Control, 2016-03) Prudêncio, Cláudia Vieira; Mantovani, Hilário Cuquetto; Cecon, Paulo Roberto; Prieto, Miguel; Vanetti, Maria Cristina Dantasantibiotics, such as nisin, generally act on Gram-positive bacteria, whereas Gram-negative bacteria exhibit natural resistance to the action of these bacteriocins. However, Gram-negative bacteria can become susceptible to the action of these antimicrobial peptides with the use of chelating agents, such as EDTA. The activity of bacteriocins can be influenced by several factors, such as environmental conditions and chemical composition of the medium. Moreover, the susceptibility of target microorganism can be altered, due to changes in cellular composition induced by environmental conditions. In this study, we examined the combined effect of variations in temperature and pH on the susceptibility of Salmonella Typhimurium to the action of nisin associated with EDTA. The reduction in the number of viable cells of Salmonella Typhimurium varied according to the temperature and pH, and major reduction in logarithmic cycles of viable cells occurred with a temperature increase. Under low temperature and low pH, a considerable reduction in the viability was also observed after prolonged exposure to bacteriocin, regardless of the presence of EDTA. According to results, temperature and pH conditions influence on the susceptibility of Salmonella Typhimurium to combination of nisin and EDTA and thus should be considered to establish optimal conditions for nisin use.Item Use of response surface methodology for optimization of xylitol production by the new yeast strain Debaryomyces hansenii UFV-170(Journal of Food Engineering, 2005-08-09) Sampaio, Fábio Coelho; Faveri, Danilo de; Mantovani, Hilario Cuquetto; Passos, Flávia M. Lopes; Perego, Patrizia; Converti, AttilioAim of this work was the optimization of xylitol production by Debaryomyces hansenii UFV-170, which already proved to be a new promising xylitol-producing yeast. Two sets of batch bioconversion tests were carried out on synthetic medium, according to two joined 3^3 and 3^2 type full factorial designs, selecting the initial xylose concentration (S0), rotational speed (v) and starting biomass concentration (X0) as independent variables and the maximum xylitol concentration (P), xylitol yield on consumed xylose (YP/S), volumetric productivity (QP) and specific productivity (qP) as response variables. The collected results were then worked out by response surface methodology (RSM). Overall optimization, conducted by overlaying the curves of the responses under investigation, allowed us to point out an optimal range of the independent variables within which the four responses were simultaneously optimized. The point chosen as representative of this optimal area corresponded to S0 = 156 g L^−1, v = 280 rpm and X0 = 6.4 g L^−1, conditions under which the model predicted P = 116.25 g L^−1, YP/S = 0.77 g g^−1, QP = 1.49 g L^−1 h^−1 and qP = 0.16 g g^−1 h^−1.Item Virtual screening of plant compounds and nonsteroidal anti-inflammatory drugs for inhibition of quorum sensing and biofilm formation in Salmonella(Microbial Pathogenesis, 2018-08) Almeida, Felipe Alves de; Vargas, Erika Lorena Giraldo; Carneiro, Deisy Guimarães; Pinto, Uelinton Manoel; Vanetti, Maria Cristina DantasSalmonella belongs to the Enterobacteriaceae family which is widely distributed in the environment due to its adaptive capacity to stress conditions. In addition, Salmonella is able to perform a type of cell-to-cell communication called quorum sensing, which leads to differential gene expression. The quorum sensing system mediated by AI-1, acyl homoserine lactones (AHLs), is incomplete in Salmonella because the luxI homolog gene, which encodes for AI-1 synthase, is missing in the genome. However, a homologue of LuxR, known as SdiA, is present and allows the detection of signaling molecules produced by other species of bacteria, leading to regulation of gene expression, mainly related to virulence and biofilm formation. Thus, in view of the importance of quorum sensing on the physiology regulation of microorganisms, the aim of the present study was to perform a virtual screening of plant compounds and nonsteroidal anti-inflammatory drugs (NASIDs) for inhibition of quorum sensing by molecular docking and biofilm formation in Salmonella. In general, most plant compounds and all NSAIDs bound in, at least, one of the three modeled structures of SdiA proteins of Salmonella Enteritidis PT4 578. In addition, many tested compounds had higher binding affinities than the AHLs and the furanones which are inducers and inhibitors of quorum sensing, respectively. The Z-phytol and lonazolac molecules were good candidates for the in vitro inhibition tests of quorum sensing mediated by AI-1 and biofilm formation in Salmonella. Thus, this study directs future prospecting of plant extracts for inhibition of quorum sensing mechanism depending on AHL and biofilm formation. In addition, the use of inhibitors of quorum sensing and biofilm formation can be combined with antibiotics for better treatment efficacy, as well as the use of these compounds to design new drugs.