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1.
J Photochem Photobiol B ; 216: 112126, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33516151

RESUMO

Prolonged exposure to Ultra Violet Radiation (UVR) adversely alters the functions of many skin cell types causing skin cancer and photoaging, which had led to increase in demand for more safe and natural sunscreens against UVR. The present study focuses on production, structural characterization and evaluation of photoprotective nature of melanin pigment derived from lime dwelling Pseudomonas sp. Melanin was characterized by solubility, UV-Vis, FT-IR, 13C-CPMAS, ESI-MS spectroscopy, including particle size, melting point and elemental analyses. In vitro cytotoxicity and photo-protective effect of Pseudomonas derived melanin (Mel-P) against UV-B (Broad Band-BB) radiations were assessed on mouse fibroblasts NIH 3 T3 cell lines. Reactive Oxygen Species (ROS) generated in NIH 3 T3 cells upon UV-B (BB) exposure was determined and quantified by Fluorescent microscopic and Flow cytometric analyses. A natural melanin obtained from Pseudomonas sp. contains 5,6- dihydroxy indole 2-carboxyic acid (DHICA) as its basic constituent and possess typical properties of eumelanin as revealed by the characterization studies. Mel-P has shown cell viability of 61.33 ± 6.58% at the concentration of 500 µg/mL proving its non-cytotoxic effect. Owing to its anti-oxidant property, melanin efficiently protected the mouse fibroblast cells from UV-B (BB) irradiation in a dose dependant manner demonstrating its potential as an active photoprotective agent.


Assuntos
Compostos de Cálcio/química , Melaninas/química , Óxidos/química , Substâncias Protetoras/química , Pseudomonas/efeitos da radiação , Neoplasias Cutâneas/prevenção & controle , Protetores Solares/química , Células 3T3 , Animais , Antioxidantes/química , Antioxidantes/metabolismo , Compostos de Cálcio/metabolismo , Fibroblastos/citologia , Fibroblastos/efeitos da radiação , Humanos , Melaninas/metabolismo , Melaninas/farmacologia , Camundongos , Óxidos/metabolismo , Substâncias Protetoras/farmacologia , Pseudomonas/metabolismo , Espécies Reativas de Oxigênio/química , Espécies Reativas de Oxigênio/metabolismo , Pele , Solubilidade , Protetores Solares/metabolismo , Raios Ultravioleta
2.
Genomics ; 111(3): 492-499, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-29530765

RESUMO

Pseudomonas frederiksbergensis ERDD5:01 is a psychrotrophic bacteria isolated from the glacial stream flowing from East Rathong glacier in Sikkim Himalaya. The strain showed survivability at high altitude stress conditions like freezing, frequent freeze-thaw cycles, and UV-C radiations. The complete genome of 5,746,824 bp circular chromosome and a plasmid of 371,027 bp was sequenced to understand the genetic basis of its survival strategy. Multiple copies of cold-associated genes encoding cold active chaperons, general stress response, osmotic stress, oxidative stress, membrane/cell wall alteration, carbon storage/starvation and, DNA repair mechanisms supported its survivability at extreme cold and radiations corroborating with the bacterial physiological findings. The molecular cold adaptation analysis in comparison with the genome of 15 mesophilic Pseudomonas species revealed functional insight into the strategies of cold adaptation. The genomic data also revealed the presence of industrially important enzymes.


Assuntos
Altitude , Genoma Bacteriano , Pseudomonas/genética , Estresse Fisiológico , Sequenciamento Completo do Genoma , Sequência de Bases , Reparo do DNA , Genômica , Estresse Oxidativo , Pseudomonas/metabolismo , Pseudomonas/fisiologia , Pseudomonas/efeitos da radiação , Raios Ultravioleta
3.
Biocontrol Sci ; 23(3): 139-143, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30249964

RESUMO

Nitrate-nitrogen (NO3-N) and nitrite-nitrogen (NO2-N) are constituents of the nitrogen cycle. NO3-N is toxic to humans, primarily due to its reduction to NO2-N. In Japan, NO3-N and NO2-N levels in tap water must not exceed 10 mg/L and only NO2-N alone not 0.04 mg/L, respectively. In this study, we verified the effect of microorganisms and ultraviolet (UV) to increase of NO2-N in water. First, all tested drinking-waters including tap water and commercial mineral water in PET bottles had < 2 mg/L NO3-N and undetectable levels (< 0.01 mg/L) of NO2-N. However, we found that NO2-N was generated in tap water left to stand at room temperature for several days, leading to increases in CF and TC counts and reduction of NO3-N. We also demonstrated that direct UV and sunlight irradiation of NO3-N-containing drinking water generated NO2-N in 1-2 h, with NO2-N reaching > 0.04 mg/mL by 4-6 h. On the other hand, NO3-N and NO2-N were undetectable in commercially purified water.


Assuntos
Água Potável/análise , Nitratos/análise , Nitritos/análise , Ciclo do Nitrogênio , Nitrogênio/química , Poluentes Químicos da Água/análise , Contagem de Colônia Microbiana , Água Potável/microbiologia , Escherichia coli/efeitos da radiação , Humanos , Japão , Oxirredução , Pseudomonas/crescimento & desenvolvimento , Pseudomonas/efeitos da radiação , Raios Ultravioleta
4.
Artigo em Inglês | MEDLINE | ID: mdl-29977868

RESUMO

Pseudomonas plecoglossicida is a facultative pathogen that is associated with diseases of multiple fish, mainly at 15-20°C. Although fish disease caused by P. plecoglossicida has led to significant economic losses, the mechanisms of the temperature-dependent virulence are unclear. Here, we identify potential pathogenicity mechanisms and demonstrate the direct regulation of several virulence factors by temperature with transcriptomic and proteomic analyses, quantitative real-time PCR (qRT-PCR), RNAi, pyoverdine (PVD) quantification, the chrome azurol S (CAS) assay, growth curve measurements, a biofilm assay, and artificial infection. The principal component analysis, the heat map generation and hierarchical clustering, together with the functional annotations of the differentially expressed genes (DEGs) demonstrated that, under different growth temperatures, the animation and focus of P. plecoglossicida are quite different, which may be the key to pathogenicity. Genes involved in PVD synthesis and in the type VI secretion system (T6SS) are specifically upregulated at the virulent temperature of 18°C. Silencing of the PVD-synthesis-related genes reduces the iron acquisition, growth, biofilm formation, distribution in host organs and virulence of the bacteria. Silencing of the T6SS genes also leads to the reduction of biofilm formation, distribution in host organs and virulence. These findings reveal that temperature regulates multiple virulence mechanisms in P. plecoglossicida, especially through iron acquisition and T6SS secretion. Meanwhile, integration of transcriptomic and proteomic data provide us with a new perspective into the pathogenesis of P. plecoglossicida, which would not have been easy to catch at either the protein or mRNA differential analyses alone, thus illustrating the power of multi-omics analyses in microbiology.


Assuntos
Perfilação da Expressão Gênica , Proteoma/análise , Infecções por Pseudomonas/veterinária , Pseudomonas/patogenicidade , Pseudomonas/efeitos da radiação , Temperatura , Regulação da Expressão Gênica/efeitos da radiação , Oligopeptídeos/biossíntese , Infecções por Pseudomonas/microbiologia , Sistemas de Secreção Tipo VI/biossíntese , Virulência/efeitos da radiação , Fatores de Virulência/biossíntese
5.
J Dairy Res ; 85(2): 232-237, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29785903

RESUMO

The present study focused on the utilisation of High Intensity Light Pulses (HILP) treatment to preserve mozzarella cheese. First, the susceptibility of Pseudomonas fluorescens and Enterobacteriaceae to HILP (fluences from 0·39 to 28·0 J/cm2) in a transparent liquid was evaluated (in-vitro tests). Afterwards, the effects on inoculated mozzarella cheese were also assessed. Then untreated (Control) and HILP treated samples were packaged and stored at 10 °C for 2 weeks. Enterobacteriaceae, Pseudomonas spp. and pH were monitored during storage. In a transparent liquid (in-vitro tests) there was a significant microbial inactivation just with 2 s of treatment. On the inoculated cheese a relevant microbial reduction of about 1 log cycle was observed, according to the exposure to the treatments. For Pseudomonas spp. in particular, in the treated samples, the microbiological acceptability limit (106 cfu/g) was never reached after 2 weeks of refrigerated storage. To sum up, the efficacy of this treatment is very interesting because a microbial reduction was observed in treated samples. HILP treatment is able to control the microbial growth and may be considered a promising way to decontaminate the surface of mozzarella cheese.


Assuntos
Queijo/microbiologia , Microbiologia de Alimentos/métodos , Conservação de Alimentos/métodos , Contagem de Colônia Microbiana , Enterobacteriaceae/citologia , Enterobacteriaceae/efeitos da radiação , Concentração de Íons de Hidrogênio , Processos Fotoquímicos , Pseudomonas/citologia , Pseudomonas/efeitos da radiação , Pseudomonas fluorescens/citologia , Pseudomonas fluorescens/efeitos da radiação
6.
Environ Sci Pollut Res Int ; 25(28): 27693-27703, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29307073

RESUMO

The importance of seawater treatment in order to avoid microbiological pollution related to aquaculture or ballast water management has increased during the last few years. Bacterial indicators used for the evaluation of different disinfection treatments are usually related with both waste and drinking water, these standards are not usual microorganisms found in seawater. Thus, it is thought necessary to study the behavior of different marine-specific organisms in regard to improve the disinfection processes in seawater. In this study, three different bacteria have been selected among major groups of bacterial community from marine waters: two water-associated, Roseobacter sp. and Pseudomonas litoralis, and one sediment-associated, Kocuria rhizophila. A kinetic inactivation model together with a post-treatment growth tendency has been obtained after the application of UV-C and UV/H2O2 processes. According to the first kinetic rate constant, different responses were obtained for the different bacterial groups. Once the treatment was applied, modeling of growth curves revealed high recover within the first 3 days after treatment, even when UV/H2O2 was applied. This study introduces a sensitivity index, in which results show different levels of resistance for both treatments, being Roseobacter sp. the most sensitive bacteria, followed by P. litoralis and K. rhizophila.


Assuntos
Desinfecção/métodos , Peróxido de Hidrogênio/química , Água do Mar/microbiologia , Raios Ultravioleta , Cinética , Micrococcaceae/efeitos dos fármacos , Micrococcaceae/efeitos da radiação , Modelos Teóricos , Processos Fotoquímicos , Pseudomonas/efeitos dos fármacos , Pseudomonas/efeitos da radiação , Roseobacter/efeitos dos fármacos , Roseobacter/efeitos da radiação
7.
PLoS One ; 12(12): e0189862, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29267321

RESUMO

Despite the overruling impact of light in the phyllosphere, little is known regarding the influence of light spectra on non-phototrophic bacteria colonizing the leaf surface. We developed an in vitro method to study phenotypic profile responses of bacterial pure cultures to different bands of the visible light spectrum using monochromatic (blue: 460 nm; red: 660 nm) and polychromatic (white: 350-990 nm) LEDs, by modification and optimization of a protocol for the Phenotype MicroArray™ technique (Biolog Inc., CA, USA). The new protocol revealed high reproducibility of substrate utilization under all conditions tested. Challenging the non-phototrophic bacterium Pseudomonas sp. DR 5-09 with white, blue, and red light demonstrated that all light treatments affected the respiratory profile differently, with blue LED having the most decisive impact on substrate utilization by impairing respiration of 140 substrates. The respiratory activity was decreased on 23 and 42 substrates under red and white LEDs, respectively, while utilization of one, 16, and 20 substrates increased in the presence of red, blue, and white LEDs, respectively. Interestingly, on four substrates contrasting utilization patterns were found when the bacterium was exposed to different light spectra. Although non-phototrophic bacteria do not rely directly on light as an energy source, Pseudomonas sp. DR 5-09 changed its respiratory activity on various substrates differently when exposed to different lights. Thus, ability to sense and distinguish between different wavelengths even within the visible light spectrum must exist, and leads to differential regulation of substrate usage. With these results, we hypothesize that different light spectra might be a hitherto neglected key stimulus for changes in microbial lifestyle and habits of substrate usage by non-phototrophic phyllospheric microbiota, and thus might essentially stratify leaf microbiota composition and diversity.


Assuntos
Luz , Pseudomonas/efeitos da radiação , Biomassa , Folhas de Planta/microbiologia , Pseudomonas/metabolismo
8.
Environ Sci Pollut Res Int ; 23(13): 13255-67, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27023813

RESUMO

Phytoremediation assisted by bacteria is seen as a promising alternative to reduce metal contamination in the environment. The main goal of this study was to characterize endophytic Pseudomonas isolated from Halimione portulacoides, a metal-accumulator plant, in salt marshes contaminated with metal(loid)s. Phylogenetic analysis based on 16S rRNA and gyrB genes showed that isolates affiliated with P. sabulinigri (n = 16), P. koreensis (n = 10), P. simiae (n = 5), P. seleniipraecipitans (n = 2), P. guineae (n = 2), P. migulae (n = 1), P. fragi (n = 1), P. xanthomarina (n = 1), and Pseudomonas sp. (n = 1). Most of these species have never been described as endophytic. The majority of the isolates were resistant to three or more metal(loid)s. Antibiotic resistance was frequent among the isolates but most likely related to species-intrinsic features. Common acquired antibiotic resistance genes and integrons were not detected. Plasmids were detected in 43.6 % of the isolates. Isolates that affiliated with different species shared the same plasmid profile but attempts to transfer metal resistance to receptor strains were not successful. Phosphate solubilization and IAA production were the most prevalent plant growth promoting traits, and 20 % of the isolates showed activity against phytopathogenic bacteria. Most isolates produced four or more extracellular enzymes. Preliminary results showed that two selected isolates promote Arabidopsis thaliana root elongation. Results highlight the diversity of endophytic Pseudomonas in H. portulacoides from contaminated sites and their potential to assist phytoremediation by acting as plant growth promoters and as environmental detoxifiers.


Assuntos
Amaranthaceae/microbiologia , Endófitos/genética , Metais Pesados/toxicidade , Pseudomonas/genética , RNA Ribossômico 16S/genética , Arabidopsis , Bactérias/genética , Biodegradação Ambiental , Farmacorresistência Bacteriana , Metais , Testes de Sensibilidade Microbiana , Filogenia , Desenvolvimento Vegetal , Raízes de Plantas/crescimento & desenvolvimento , Plasmídeos/genética , Pseudomonas/enzimologia , Pseudomonas/efeitos da radiação , Cloreto de Sódio , Áreas Alagadas
9.
Can J Microbiol ; 61(7): 457-66, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25996177

RESUMO

Common problems in a windowfarm system (a vertical and indoor hydroponic system) are phytopathogen infections in plants and excessive buildup of biofilms. The objectives of this study were (i) to promote plant health by making plants more resistant to infection by using beneficial biosurfactant-producing Pseudomonas chlororaphis around the roots and (ii) to minimize biofilm buildup by ultraviolet (UV) irradiation of the water reservoir, thereby extending the lifespan of the whole system with minimal maintenance. Pseudomonas chlororaphis-treated lettuce grew significantly better than nontreated lettuce, as indicated by enhancement of color, mass, length, and number of leaves per head (p < 0.05). The death rate of the lettuce was reduced by ∼ 50% when the lettuce was treated with P. chlororaphis. UV irradiation reduced the bacteria (4 log reduction) and algae (4 log reduction) in the water reservoirs and water tubing systems. Introduction of P. chlororaphis into the system promoted plant growth and reduced damage caused by the plant pathogen Pythium ultimum. UV irradiation of the water reservoir reduced algal and biofilm growth and extended the lifespan of the system.


Assuntos
Biofilmes/efeitos da radiação , Lactuca/crescimento & desenvolvimento , Pseudomonas/fisiologia , Biofilmes/crescimento & desenvolvimento , Lactuca/microbiologia , Lactuca/efeitos da radiação , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/microbiologia , Folhas de Planta/efeitos da radiação , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/microbiologia , Raízes de Plantas/efeitos da radiação , Pseudomonas/efeitos da radiação , Raios Ultravioleta
10.
Meat Sci ; 98(3): 383-91, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25042241

RESUMO

The objectives of the present work were to assess the use of moderate doses of gamma irradiation (2 to 5 kGy) and to reduce the risk of pathogen presence without altering the quality attributes of bovine trimmings and of patties made of irradiated trimmings. Microbiological indicators (coliforms, Pseudomonas spp and mesophilic aerobic counts), physicochemical indicators (pH, color and tiobarbituric acid) and sensory changes were evaluated during storage. 5 kGy irradiation doses slightly increased off flavors in patties. Two pathogenic markers (Listeria monocytogenes and Escherichia coli O157:H7) were inoculated at high or low loads to trimming samples which were subsequently irradiated and lethality curves were obtained. Provided that using irradiation doses ≤2.5 kGy are used, reductions of 2 log CFU/g of L. monocytogenes and 5 log CFU/g of E. coli O157:H7 are expected. It seems reasonable to suppose that irradiation can be successfully employed to improve the safety of frozen trimmings when initial pathogenic bacteria burdens are not extremely high.


Assuntos
Qualidade de Produtos para o Consumidor , Escherichia coli O157/efeitos da radiação , Manipulação de Alimentos/métodos , Microbiologia de Alimentos , Raios gama , Listeria monocytogenes/efeitos da radiação , Carne/microbiologia , Animais , Bovinos , Contagem de Colônia Microbiana , Humanos , Carne/análise , Carne/normas , Produtos da Carne/análise , Produtos da Carne/microbiologia , Produtos da Carne/normas , Pseudomonas/efeitos da radiação
11.
Appl Microbiol Biotechnol ; 98(10): 4671-81, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24522726

RESUMO

Sulfamethoxazole is a common antibiotic that is frequently detected in wastewater and surface water. This study investigated the biodegradation and metabolic pathway of sulfamethoxazole by Pseudomonas psychrophila HA-4, a cold-adapted bacterium. Strain HA-4, which uses sulfamethoxazole as its sole source of carbon and energy, was isolated at a low temperature (10 °C) and identified as P. psychrophila by physico-biochemical characterization and 16S rRNA gene sequence analysis. Strain HA-4 removed sulfamethoxazole at temperatures ranging from 5.0 °C to 30 °C, with the maximal removal rate at 10 °C. The maximal removal rate of sulfamethoxazole by strain HA-4 was 34.30 % after 192 h at 10 °C. The highest percentage of unsaturated fatty acid was determined to be 23.03 % at 10 °C, which adheres to the characteristic for cold-adapted psychrophiles and psychrotrophs. At low concentrations of sulfamethoxazole, the growth kinetics correlated well with the Haldane model. The single-substrate parameter values of sulfamethoxazole on cell growth were determined to be µ max = 0.01 h(-1), K s = 20.91 mg/l and K i = 170.60 mg/l. Additionally, the major intermediates from sulfamethoxazole biodegradation by strain HA-4, including aniline, 3-amino-5-methylisoxazole, 4-aminothiophenol and sulfanilamide, were identified by GC-MS and high-resolution mass spectrometry (HR-MS) analysis. The results demonstrate that strain HA-4 has the potential to degrade sulfamethoxazole at low temperatures.


Assuntos
Redes e Vias Metabólicas/genética , Pseudomonas/isolamento & purificação , Pseudomonas/metabolismo , Sulfametoxazol/metabolismo , Biotransformação , Carbono/metabolismo , Análise por Conglomerados , Temperatura Baixa , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Metabolismo Energético , Ácidos Graxos Insaturados/análise , Espectrometria de Massas , Microscopia Eletrônica de Varredura , Dados de Sequência Molecular , Filogenia , Pseudomonas/classificação , Pseudomonas/efeitos da radiação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
12.
Photochem Photobiol Sci ; 12(6): 974-86, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23493991

RESUMO

The dose-dependent variation of oxidative cellular damage imposed by UVB exposure in a representative estuarine bacterial strain, Pseudomonas sp. NT5I1.2B, was studied at different growth phases (mid-exponential, late-exponential, and stationary), growth temperatures (15 °C and 25 °C) and growth media (nutrient-rich Tryptic Soy Broth [TSB] and nutrient-poor M9). Survival and markers of oxidative damage (lipid peroxidation, protein carbonylation, DNA strand breakage, and DNA-protein cross-links) were monitored during exposure to increasing UVB doses (0-60 kJ m(-2)). Oxidative damage did not follow a clear linear dose-dependent pattern, particularly at high UVB doses (>10 kJ m(-2)), suggesting a dynamic interaction between damage induction and repair during irradiation and/or saturation of oxidative damage. Survival of stationary phase cells generally exceeded that of exponential phase cells by up to 33.5 times; the latter displayed enhanced levels of DNA-protein cross-links (up to 15.6-fold) and protein carbonylation (up to 6.0-fold). Survival of mid-exponential phase cells was generally higher at 15 °C than at 25 °C (up to 6.6-fold), which was accompanied by lower levels of DNA strand breaks (up to 4000-fold), suggesting a temperature effect on reactive oxygen species (ROS) generation and/or ROS interaction with cellular targets. Survival under medium-high UVB doses (>10 kJ m(-2)) was generally higher (up to 5.4-fold) in cells grown in TSB than in M9. These results highlight the influence of growth conditions preceding irradiation on the extent of oxidative damage induced by UVB exposure in bacteria.


Assuntos
Pseudomonas/crescimento & desenvolvimento , Pseudomonas/efeitos da radiação , Proteínas de Bactérias/metabolismo , Quebras de DNA/efeitos da radiação , DNA Bacteriano/genética , DNA Bacteriano/metabolismo , Peroxidação de Lipídeos/efeitos da radiação , Estresse Oxidativo , Pseudomonas/genética , Pseudomonas/metabolismo , Raios Ultravioleta
13.
Photochem Photobiol ; 89(3): 640-8, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23360113

RESUMO

The purpose of this study was to explore the possible link between metals and UV-B-induced damage in bacteria. The effect of growth in the presence of enhanced concentrations of different transition metals (Co, Cu, Fe, Mn and Zn) on the UV-B sensitivity of a set of bacterial isolates was explored in terms of survival, activity and oxidative stress biomarkers (ROS generation, damage to DNA, lipid and proteins and activity of antioxidant enzymes). Metal amendment, particularly Fe, Cu and Mn, enhanced bacterial inactivation during irradiation by up to 35.8%. Amendment with Fe increased ROS generation during irradiation by 1.2-13.3%, DNA damage by 10.8-37.4% and lipid oxidative damage by 9.6-68.7%. Lipid damage during irradiation also increased after incubation with Cu and Co by up to 66.8% and 56.5% respectively. Mn amendment decreased protein carbonylation during irradiation by up to 44.2%. These results suggest a role of Fe, Co, Cu and Mn in UV-B-induced bacterial inactivation and the importance of metal homeostasis to limit the detrimental effects of ROS generated during irradiation.


Assuntos
Micrococcus/efeitos dos fármacos , Paracoccus/efeitos dos fármacos , Pseudomonas/efeitos dos fármacos , Staphylococcus/efeitos dos fármacos , Elementos de Transição/farmacologia , Catalase/metabolismo , Viabilidade Microbiana/efeitos dos fármacos , Viabilidade Microbiana/efeitos da radiação , Micrococcus/crescimento & desenvolvimento , Micrococcus/metabolismo , Micrococcus/efeitos da radiação , Oxirredução , Estresse Oxidativo , Paracoccus/crescimento & desenvolvimento , Paracoccus/metabolismo , Paracoccus/efeitos da radiação , Carbonilação Proteica , Pseudomonas/crescimento & desenvolvimento , Pseudomonas/metabolismo , Pseudomonas/efeitos da radiação , Espécies Reativas de Oxigênio , Staphylococcus/crescimento & desenvolvimento , Staphylococcus/metabolismo , Staphylococcus/efeitos da radiação , Superóxido Dismutase/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Raios Ultravioleta
14.
Appl Biochem Biotechnol ; 169(5): 1467-81, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23315264

RESUMO

The objective of the study was to execute mutant bacteria for efficient biodegradation of sulfonated azo dye, Green HE4B (GHE4B). UV irradiation was used to introduce random mutations in Pseudomonas sp. LBC1. Genetic alterations induced by UV irradiation in selected mutant bacteria were confirmed by random amplification of polymorphic DNA technique. The mutant bacteria named as Pseudomonas sp. 1 F reduced the time required for complete degradation of recalcitrant dye GHE4B by 25 % when compared with the wild one. The biodegradation was monitored by UV-Vis spectrophotometric analysis. Activities of enzymes like laccase, lignin peroxidase, veratryl alcohol oxidase, and NADH dichlorophenol indophenol reductase were found to be boosted in mutant bacteria as a consequence of UV-induced mutation. Matrix-assisted laser desorption/ionization-time of flight analysis of differentially expressed proteins of mutant bacteria suggested active role of antioxidant enzymes in the degradation of the dye. The degradation product was analyzed by Fourier transform infrared spectroscopy, high-performance thin-layer chromatography, and gas chromatography-mass spectrometry. Results revealed few variations in the degradation end products of wild-type and mutant bacteria. Phytotoxicity study underlined the safer biodegradation of GHE4B by mutant Pseudomonas sp. 1 F.


Assuntos
Proteínas de Bactérias/química , Corantes/metabolismo , Pseudomonas/enzimologia , Pseudomonas/genética , Triazinas/metabolismo , Oxirredutases do Álcool/química , Biodegradação Ambiental , Meios de Cultivo Condicionados/farmacologia , Ativação Enzimática , Cinética , Lacase/química , Mutagênese , Peroxidases/química , Pseudomonas/efeitos da radiação , Quinona Redutases/química , Técnica de Amplificação ao Acaso de DNA Polimórfico , Sorghum/efeitos dos fármacos , Sorghum/crescimento & desenvolvimento , Raios Ultravioleta
15.
Chemosphere ; 90(4): 1348-58, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22980957

RESUMO

The influence of low dose irradiation pretreatment on the microbial decolouration and degradation of Reactive Red-120 (RR-120) dye was investigated in detail by using Pseudomonas sp. SUK1. About 27%, 56% and 66% decolouration of 150 ppm RR-120 dye solution was observed by applying 0, 0.5 and 1 kGy doses, respectively, in the first step followed by microbial treatment for 24 h under static condition. Similarly, about 70%, 88% and 90% TOC removal was observed by applying 0, 0.5 and 1 kGy doses, respectively, in the first step followed by the microbial treatment for 96 h under static condition. The radiation induced fragmented products of RR-120 at doses of 0.5 and 1 kGy were investigated by FTIR and electrospray ionization-MS analysis. The induction of the enzymes viz. laccase, tyrosinase, azoreductase and NADH-2,6-dichlorophenol indophenol reductase was studied in the decolourised solution obtained after irradiating 150 ppm RR-120 dye solution with 0 and 1 kGy doses followed by the microbial treatment for 96 h under static condition. The enzymatic degradation products were studied by FTIR, HPLC and GC-MS. The toxicity study of the treated dye solution on plants revealed the degradation of RR-120 into non-toxic products by combined radiation-microbial treatment. This study explores a reliable and promising way to use industrially viable dose (≤1 kGy) and microbial strain viz. Pseudomonas sp. SUK1 for permissible safe disposal of dye solutions from textile industries.


Assuntos
Recuperação e Remediação Ambiental/métodos , Resíduos Industriais/análise , Pseudomonas/efeitos da radiação , Indústria Têxtil , Triazinas/metabolismo , Poluentes Químicos da Água/metabolismo , Biodegradação Ambiental , Cromatografia Líquida de Alta Pressão , Raios gama , Pseudomonas/metabolismo , Doses de Radiação , Espectroscopia de Infravermelho com Transformada de Fourier , Triazinas/análise , Triazinas/química , Triazinas/efeitos da radiação , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/química , Poluentes Químicos da Água/efeitos da radiação
16.
J Bacteriol ; 194(20): 5721, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23012293

RESUMO

Pseudomonas psychrophila HA-4 is a cold-adaptable, sulfamethoxazole-degrading bacterium. The genes related to its cold adaptation mechanism and sulfamethoxazole metabolism were unknown. We present the draft genome of strain HA-4. It could provide further insight into the sulfamethoxazole-degrading mechanism of strain HA-4.


Assuntos
DNA Bacteriano/química , DNA Bacteriano/genética , Genoma Bacteriano , Pseudomonas/genética , Análise de Sequência de DNA , Adaptação Fisiológica , Temperatura Baixa , Redes e Vias Metabólicas/genética , Dados de Sequência Molecular , Pseudomonas/isolamento & purificação , Pseudomonas/fisiologia , Pseudomonas/efeitos da radiação , Sulfametoxazol/metabolismo
17.
Pol J Microbiol ; 60(2): 119-23, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21905628

RESUMO

The efficacy of UV treatment to control bacterial adhesion onto hard surfaces was investigated in laboratory conditions. The major characteristics necessary for biofilm formation like extracellular polymeric substance (EPS) production, carbohydrate and protein concentration in EPS, and adhesion ability onto hard surface were studied using two bacterial strains isolated from marine biofilms. The results showed that there was a considerable difference between the control and UV treated bacterial cultures in their viability, production of EPS, and adhesion ability. The protein and carbohydrate concentration of the EPS and the adhesion of bacterial cells to surface were also considerably reduced due to UV treatment. This study indicates that treatment of water with UV light may be used to control biofilm development on hard surfaces.


Assuntos
Alteromonas/efeitos da radiação , Aderência Bacteriana/efeitos da radiação , Biofilmes/efeitos da radiação , Pseudomonas/efeitos da radiação , Raios Ultravioleta , Alteromonas/crescimento & desenvolvimento , Alteromonas/fisiologia , Análise de Variância , Aderência Bacteriana/fisiologia , Biofilmes/crescimento & desenvolvimento , Biopolímeros/biossíntese , Biopolímeros/efeitos da radiação , Espaço Extracelular/química , Vidro , Pseudomonas/crescimento & desenvolvimento , Pseudomonas/fisiologia
18.
Mol Plant ; 4(3): 516-26, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21447757

RESUMO

The Arabidopsis accelerated cell death 6-1 (acd6-1) mutant shows constitutive defense, cell death, and extreme dwarf phenotypes. In a screen for acd6-1 suppressors, we identified a mutant that was disrupted by a T-DNA in the PHOSPHATE TRANSPORTER 4;1 (PHT4;1) gene. The suppressor mutant pht4;1-1 is dominant, expresses truncated PHT4;1 transcripts, and is more susceptible to virulent Pseudomonas syringae strains but not to several avirulent strains. Treatment with a salicylic acid (SA) agonist induced a similar level of resistance in Col-0 and pht4;1-1, suggesting that PHT4;1 acts upstream of the SA pathway. Genetic analysis further indicates that PHT4;1 contributes to SID2-dependent and -independent pathways. Transgenic expression of the DNA fragment containing the PHT4;1-1 region or the full-length PHT4;1 gene in wild-type conferred enhanced susceptibility to Pseudomonas infection. Interestingly, expression of PHT4;1 is regulated by the circadian clock. Together, these data suggest that the phosphate transporter PHT4;1 is critical for basal defense and also implicate a potential role of the circadian clock in regulating innate immunity of Arabidopsis.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/imunologia , Relógios Circadianos , Proteínas de Transporte de Fosfato/metabolismo , Anquirinas/metabolismo , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Arabidopsis/microbiologia , Proteínas de Arabidopsis/genética , Relógios Circadianos/efeitos dos fármacos , Relógios Circadianos/genética , Relógios Circadianos/efeitos da radiação , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Genes Dominantes/genética , Luz , Fenótipo , Proteínas de Transporte de Fosfato/genética , Pseudomonas/efeitos dos fármacos , Pseudomonas/patogenicidade , Pseudomonas/efeitos da radiação , Ácido Salicílico/farmacologia , Supressão Genética/efeitos dos fármacos , Supressão Genética/efeitos da radiação , Virulência/efeitos dos fármacos , Virulência/efeitos da radiação
19.
Appl Environ Microbiol ; 77(2): 460-70, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21115713

RESUMO

The refrigerated storage of raw milk throughout the dairy chain prior to heat treatment creates selective conditions for growth of psychrotolerant bacteria. These bacteria, mainly belonging to the genus Pseudomonas, are capable of producing thermoresistant extracellular proteases and lipases, which can cause spoilage and structural defects in pasteurized and ultra-high-temperature-treated milk (products). To map the influence of refrigerated storage on the growth of these pseudomonads, milk samples were taken after the first milking turn and incubated laboratory scale at temperatures simulating optimal and suboptimal preprocessing storage conditions. The outgrowth of Pseudomonas members was monitored over time by means of cultivation-independent denaturing gradient gel electrophoresis (DGGE). Isolates were identified by a polyphasic approach. These incubations revealed that outgrowth of Pseudomonas members occurred from the beginning of the dairy chain (farm tank) under both optimal and suboptimal storage conditions. An even greater risk for outgrowth, as indicated by a vast increase of about 2 log CFU per ml raw milk, existed downstream in the chain, especially when raw milk was stored under suboptimal conditions. This difference in Pseudomonas outgrowth between optimal and suboptimal storage was already statistically significant within the farm tank. The predominant taxa were identified as Pseudomonas gessardii, Pseudomonas gessardii-like, Pseudomonas fluorescens-like, Pseudomonas lundensis, Pseudomonas fragi, and Pseudomonas fragi-like. Those taxa show an important spoilage potential as determined on elective media for proteolysis and lipolysis.


Assuntos
Biodiversidade , Viabilidade Microbiana , Leite/microbiologia , Pseudomonas/crescimento & desenvolvimento , Pseudomonas/efeitos da radiação , Refrigeração , Animais , Carga Bacteriana , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Eletroforese em Gel de Poliacrilamida , Dados de Sequência Molecular , Desnaturação de Ácido Nucleico , Filogenia , Pseudomonas/classificação , Pseudomonas/genética , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
20.
J Gen Appl Microbiol ; 57(6): 379-86, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22353743

RESUMO

Microorganisms able to bioconvert DL-2-amino-Δ(2)-thiazoline-4-carboxylic acid (DL-ATC) into L-cysteine were originally isolated from 10 soil samples with DL-ATC as the sole nitrogen source. Ninety-seven L-cysteine-producing bacterial strains were screened out and obtained in pure culture. Among them, a strain, designated as HUT-78, was selected as the best producer, with a molar bioconversion rate of 60%. Based on the 16S rRNA gene sequence analysis, this isolate was placed within the genus Pseudomonas. A novel mutant of this strain with a significantly reduced activity of L-cysteine desulfhydrase, a L-cysteine-decomposing enzyme, was derived by UV-mutagenesis. This mutant, designated as mHUT-78, exhibited a 42% increase in L-cysteine producing activity. Moreover, the bioconversion reactions in both the parent and the mutant strain were significantly accelerated by co-overexpression of the two key enzymes, AtcB and AtcC, involved in the bioconversion reaction.


Assuntos
Cisteína/metabolismo , Engenharia Metabólica , Redes e Vias Metabólicas/genética , Pseudomonas/isolamento & purificação , Pseudomonas/metabolismo , Tiazóis/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biotransformação , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Expressão Gênica , Mutação , Nitrogênio/metabolismo , Pseudomonas/genética , Pseudomonas/efeitos da radiação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Microbiologia do Solo , Raios Ultravioleta
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