RESUMO
Tellurium oxyanions are chemical species of great toxicity and their presence in the environment has increased because of mining industries and photovoltaic and electronic waste. Recovery strategies for this metalloid that are based on micro-organisms are of interest, but further studies of the transport systems and enzymes responsible for implementing tellurium transformations are required because many mechanisms remain unknown. Here, we investigated the involvement in tellurite uptake of the putative phosphate transporter PitB (PP1373) in soil bacterium Pseudomonas putida KT2440. For this purpose, through a method based on the CRISPR/Cas9 system, we generated a strain deficient in the pitB gene and characterized its phenotype on exposing it to varied concentrations of tellurite. Growth curves and transmission electronic microscopy experiments for the wild-type and ΔpitB strains showed that both were able to internalize tellurite into the cytoplasm and reduce the oxyanion to black nano-sized and rod-shaped tellurium particles, although the ΔpitB strain showed an increased resistance to the tellurite toxic effects. At a concentration of 100 µM tellurite, where the biomass formation of the wild-type strain decreased by half, we observed a greater ability of ΔpitB to reduce this oxyanion with respect to the wild-type strain (~38 vs ~16â%), which is related to the greater biomass production of ΔpitB and not to a greater consumption of tellurite per cell. The phenotype of the mutant was restored on over-expressing pitB in trans. In summary, our results indicate that PitB is one of several transporters responsible for tellurite uptake in P. putida KT2440.
Assuntos
Proteínas de Bactérias/metabolismo , Proteínas de Transporte de Fosfato/metabolismo , Pseudomonas putida/metabolismo , Telúrio/metabolismo , Proteínas de Bactérias/genética , Transporte Biológico , Biomassa , Biotransformação , Mutação , Nanoestruturas/química , Nanoestruturas/toxicidade , Proteínas de Transporte de Fosfato/genética , Pseudomonas putida/efeitos dos fármacos , Pseudomonas putida/crescimento & desenvolvimento , Telúrio/química , Telúrio/toxicidadeRESUMO
Objective. To conduct a health impact assessment (HIA) to quantify health benefits for several PM and O3 air pollution reduction scenarios in the Mexico City Metropolitan Area (MCMA). Results from this HIA will contribute to the scientific support of the MCMA air quality management plan (PROAIRE) for the period 2011-2020. Materials and methods. The HIA methodology consisted of four steps: 1) selection of the air pollution reduction scenarios, 2) identification of the at-risk population and health outcomes for the 2005 baseline scenario, 3) selection of concentration-response functions and 4) estimation of health impacts. Results. Reductions of PM10 levels to 20 μg/m³ and O3 levels to 0.050ppm (98 µg/m³) would prevent 2300 and 400 annual deaths respectively. The greatest health impact was seen in the over-65 age group and in mortality due to cardiopulmonary and cardiovascular disease. Conclusion. Improved air quality in the MCMA could provide significant health benefits through focusing interventions by exposure zones.
Objetivo. Realizar una evaluación de impacto en salud (EIS) que documente los beneficios en salud ante diversos escenarios de reducción de PM10 y O3 en el aire de la Zona Metropolitana del Valle de México (ZMVM). Los resultados contribuyen al sustento científico del plan de gestión de calidad del aire (PROAIRE 2011-2020). Material y métodos. La metodología de EIS comprende cuatro pasos: 1) selección de los escenarios de reducción, 2) identificación de la población en riesgo y de los eventos en salud para el año basal 2005, 3) selección de las funciones de concentración-respuesta y 4) estimación del impacto en la salud. Resultados. Reducciones de PM10 a 20μg/m³ y de O3 a 0.050ppm (98 µg/m³) evitarían, respectivamente, cerca de 2 300 y 400 muertes por año. El mayor impacto se observa en el grupo de más de 65 años y en la mortalidad por causas cardiopulmonares y cardiovasculares. Conclusiones. Mejorar la calidad del aire en la ZMVM podría reflejar importantes beneficios para la salud focalizados por zonas o áreas de exposición.
Assuntos
Pseudomonas putida/metabolismo , Estirenos/metabolismo , Aldeído Oxirredutases/metabolismo , Biodegradação Ambiental , Compostos de Epóxi/metabolismo , Proteínas de Escherichia coli , Ácido Glutâmico/metabolismo , Isomerases/metabolismo , Oxirredução , Consumo de Oxigênio , Fenilacetatos/metabolismo , Pseudomonas putida/enzimologia , Pseudomonas putida/crescimento & desenvolvimento , Estireno , Succinatos/metabolismo , Ácido SuccínicoRESUMO
The plant growth-promoting rhizobacterial strain Pseudomonas putida Rs-198 was isolated from salinized soils from Xinjiang Province. We optimized the composition of the low-cost medium of P. putida Rs-198 based on its bacterial concentration, as well as its phosphate-dissolving and indole acetic acid (IAA)-producing capabilities using the response surface methodology (RSM), and a mathematical model was developed to show the effect of each medium component and its interactions on phosphate dissolution and IAA production. The model predicted a maximum phosphate concentration in medium containing 63.23 mg/L inorganic phosphate with 49.22 g/L corn flour, 14.63 g/L soybean meal, 2.03 g/L K2HPO4, 0.19 g/L MnSO4 and 5.00 g/L NaCl. The maximum IAA concentration (18.73 mg/L) was predicted in medium containing 52.41 g/L corn flour, 15.82 g/L soybean meal, 2.40 g/L K2HPO4, 0.17 g/L MnSO4 and 5.00 g/L NaCl. These predicted values were also verified through experiments, with a cell density of 10(13) cfu/mL, phosphate dissolution of 64.33 mg/L, and IAA concentration of 18.08 mg/L. The excellent correlation between predicted and measured values of each model justifies the validity of both the response models. The study aims to provide a basis for industrialized fermentation using P. putida Rs-198.
Assuntos
Meios de Cultura/química , Pseudomonas putida/crescimento & desenvolvimento , Carga Bacteriana , Custos e Análise de Custo , Meios de Cultura/economia , Ácidos Indolacéticos/metabolismo , Programas de Rastreamento , Modelos Teóricos , Fosfatos/metabolismo , Pseudomonas putida/metabolismoRESUMO
The plant growth-promoting rhizobacterial strain Pseudomonas putida Rs-198 was isolated from salinized soils from Xinjiang Province. We optimized the composition of the low-cost medium of P. putida Rs-198 based on its bacterial concentration, as well as its phosphate-dissolving and indole acetic acid (IAA)-producing capabilities using the response surface methodology (RSM), and a mathematical model was developed to show the effect of each medium component and its interactions on phosphate dissolution and IAA production. The model predicted a maximum phosphate concentration in medium containing 63.23 mg/L inorganic phosphate with 49.22 g/L corn flour, 14.63 g/L soybean meal, 2.03 g/L K2HPO4, 0.19 g/L MnSO4 and 5.00 g/L NaCl. The maximum IAA concentration (18.73 mg/L) was predicted in medium containing 52.41 g/L corn flour, 15.82 g/L soybean meal, 2.40 g/L K2HPO4, 0.17 g/L MnSO4 and 5.00 g/L NaCl. These predicted values were also verified through experiments, with a cell density of 10(13) cfu/mL, phosphate dissolution of 64.33 mg/L, and IAA concentration of 18.08 mg/L. The excellent correlation between predicted and measured values of each model justifies the validity of both the response models. The study aims to provide a basis for industrialized fermentation using P. putida Rs-198.
Assuntos
Meios de Cultura/química , Pseudomonas putida/crescimento & desenvolvimento , Carga Bacteriana , Custos e Análise de Custo , Meios de Cultura/economia , Ácidos Indolacéticos/metabolismo , Programas de Rastreamento , Modelos Teóricos , Fosfatos/metabolismo , Pseudomonas putida/metabolismoRESUMO
Temporomandibular disorder (TMD) is a common condition. This study is part of a research group and it investigated the prevalence of TMD and myofascial pain and its association with gender, age and socioeconomic class. The sample comprised 100 subjects, aged 15 to 70, users of the Family Health Units' services, in the city of Recife, PE, Brazil. The TMD degree was evaluated using the Research Diagnostic Criteria for TMD and socioeconomic class by the Economic Classification Criteria Brazil. Categorical variables were analyzed by chi-square test for proportions and Fisher's exact test for 2x2 tables, and binary logistic analysis to track the relationship between the independent and dependent variables. According to the results, 42% of the subjects had TMD and 14% myofascial pain. No statistically significant association could be found between TMD and gender or socioeconomic class, but it was found to have statistically significant association with age, and myofascial pain was associated with socioeconomic class. Considering that the results of the present study should be confirmed by further studies and the fact that this was a pilot study, the prevalence must be analyzed with caution.
Disfunção temporomandibular (DTM) é uma condição comum. Este estudo é parte de um grupo de pesquisa e investigou a prevalência de DTM e dor miofascial e suas associações com sexo, idade e classe socioeconômica. A amostra foi composta por 100 indivíduos, com idades entre 15 e 70 anos, usuários das Unidades de Saúde da Família, na cidade de Recife, PE. O grau de DTM foi avaliado usando os Critérios de Diagnósticos Científicos em DTM, e classe socioeconômica com o Critério de Classificação Econômica Brasil. As variáveis categóricas foram analisadas pelo teste do qui-quadrado para proporções e teste exato de Fisher para tabelas 2x2, e a análise logística binária para traçar a relação entre as variáveis independentes e dependentes. De acordo com os resultados, 42% dos indivíduos tinham DTM e 14% dor miofascial. Não houve associação estatisticamente significativa entre DTM e sexo ou classe socioeconômica, mas houve associação estatisticamente significativa com a idade e a dor miofascial foi associada com a classe socioeconômica. Considerando-se que os resultados do presente estudo devam ser confirmados em outros estudos e por causa de sua natureza piloto, a prevalência deve ser analisada com cautela.
Assuntos
Regulação Bacteriana da Expressão Gênica , Óperon , Fenilacetatos/metabolismo , Pseudomonas putida/genética , Pseudomonas putida/crescimento & desenvolvimento , Técnicas de Cultura de Células , Divisão Celular , Meios de Cultura , Carbono/metabolismo , Coenzima A Ligases/biossíntese , Coenzima A Ligases/genética , Compostos Inorgânicos/metabolismo , Oxigenases/biossíntese , Oxigenases/genética , Pseudomonas putida/metabolismo , Reprodutibilidade dos Testes , Ácido Succínico/metabolismo , Ativação TranscricionalRESUMO
Growth associated biosynthesis of medium chain length poly-3-hydroxyalkanoates (mcl-PHA) in Pseudomonas putida Bet001 isolated from palm oil mill effluent was studied. Models with substrate inhibition terms described well the kinetics of its growth. Selected fatty acids (C8:0 to C18:1) and ammonium were used as carbon and nitrogen sources during growth and PHA biosynthesis, resulting in PHA accumulation of about 50 to 69% (w/w) and PHA yields ranging from 10.12 g L(-1) to 15.45 g L(-1), respectively. The monomer composition of the PHA ranges from C4 to C14, and was strongly influenced by the type of carbon substrate fed. Interestingly, an odd carbon chain length (C7) monomer was also detected when C18:1 was fed. Polymer showed melting temperature (T m) of 42.0 (± 0.2) °C, glass transition temperature (T g) of -1.0 (± 0.2) °C and endothermic melting enthalpy of fusion (ΔHf) of 110.3 (± 0.1) J g(-1). The molecular weight (M w) range of the polymer was relatively narrow between 55 to 77 kDa.
Assuntos
Carbono/metabolismo , Poli-Hidroxialcanoatos/metabolismo , Pseudomonas putida/crescimento & desenvolvimento , Pseudomonas putida/metabolismo , Compostos de Amônio/metabolismo , Meios de Cultura/química , Microbiologia Ambiental , Ácidos Graxos/metabolismo , Microbiologia Industrial , Resíduos Industriais , Pseudomonas putida/isolamento & purificação , TemperaturaRESUMO
Growth associated biosynthesis of medium chain length poly-3-hydroxyalkanoates (mcl-PHA) in Pseudomonas putida Bet001 isolated from palm oil mill effluent was studied. Models with substrate inhibition terms described well the kinetics of its growth. Selected fatty acids (C8:0 to C18:1) and ammonium were used as carbon and nitrogen sources during growth and PHA biosynthesis, resulting in PHA accumulation of about 50 to 69% (w/w) and PHA yields ranging from 10.12 g L-1 to 15.45 g L-1, respectively. The monomer composition of the PHA ranges from C4 to C14, and was strongly influenced by the type of carbon substrate fed. Interestingly, an odd carbon chain length (C7) monomer was also detected when C18:1 was fed. Polymer showed melting temperature (Tm) of 42.0 (± 0.2) °C, glass transition temperature (Tg) of -1.0 (± 0.2) °C and endothermic melting enthalpy of fusion (ΔHf) of 110.3 (± 0.1) J g-1. The molecular weight (Mw) range of the polymer was relatively narrow between 55 to 77 kDa.
Assuntos
Carbono/metabolismo , Poli-Hidroxialcanoatos/metabolismo , Pseudomonas putida/crescimento & desenvolvimento , Pseudomonas putida/metabolismo , Compostos de Amônio/metabolismo , Meios de Cultura/química , Microbiologia Ambiental , Ácidos Graxos/metabolismo , Microbiologia Industrial , Resíduos Industriais , Pseudomonas putida/isolamento & purificação , TemperaturaRESUMO
Growth associated biosynthesis of medium chain length poly-3-hydroxyalkanoates (mcl-PHA) in Pseudomonas putida Bet001 isolated from palm oil mill effluent was studied. Models with substrate inhibition terms described well the kinetics of its growth. Selected fatty acids (C8:0 to C18:1) and ammonium were used as carbon and nitrogen sources during growth and PHA biosynthesis, resulting in PHA accumulation of about 50 to 69% (w/w) and PHA yields ranging from 10.12 g L-1 to 15.45 g L-1, respectively. The monomer composition of the PHA ranges from C4 to C14, and was strongly influenced by the type of carbon substrate fed. Interestingly, an odd carbon chain length (C7) monomer was also detected when C18:1 was fed. Polymer showed melting temperature (Tm) of 42.0 (± 0.2) °C, glass transition temperature (Tg) of -1.0 (± 0.2) °C and endothermic melting enthalpy of fusion (ΔHf) of 110.3 (± 0.1) J g-1. The molecular weight (Mw) range of the polymer was relatively narrow between 55 to 77 kDa.
Assuntos
Carbono/metabolismo , Poli-Hidroxialcanoatos/metabolismo , Pseudomonas putida/crescimento & desenvolvimento , Pseudomonas putida/metabolismo , Compostos de Amônio/metabolismo , Meios de Cultura/química , Microbiologia Ambiental , Ácidos Graxos/metabolismo , Microbiologia Industrial , Resíduos Industriais , Pseudomonas putida/isolamento & purificação , TemperaturaRESUMO
The plant growth-promoting rhizobacterial strain Pseudomonas putida Rs-198 was isolated from salinized soils from Xinjiang Province. We optimized the composition of the low-cost medium of P. putida Rs-198 based on its bacterial concentration, as well as its phosphate-dissolving and indole acetic acid (IAA)-producing capabilities using the response surface methodology (RSM), and a mathematical model was developed to show the effect of each medium component and its interactions on phosphate dissolution and IAA production. The model predicted a maximum phosphate concentration in medium containing 63.23 mg/L inorganic phosphate with 49.22 g/L corn flour, 14.63 g/L soybean meal, 2.03 g/L K2HPO4, 0.19 g/L MnSO4 and 5.00 g/L NaCl. The maximum IAA concentration (18.73 mg/L) was predicted in medium containing 52.41 g/L corn flour, 15.82 g/L soybean meal, 2.40 g/L K2HPO4, 0.17 g/L MnSO4 and 5.00 g/L NaCl. These predicted values were also verified through experiments, with a cell density of 10(13) cfu/mL, phosphate dissolution of 64.33 mg/L, and IAA concentration of 18.08 mg/L. The excellent correlation between predicted and measured values of each model justifies the validity of both the response models. The study aims to provide a basis for industrialized fermentation using P. putida Rs-198.
Assuntos
Meios de Cultura/química , Pseudomonas putida/crescimento & desenvolvimento , Carga Bacteriana , Custos e Análise de Custo , Meios de Cultura/economia , Ácidos Indolacéticos/metabolismo , Programas de Rastreamento , Modelos Teóricos , Fosfatos/metabolismo , Pseudomonas putida/metabolismoRESUMO
In situ biosurfactant (rhamnolipid) production by Pseudomonas putida CB-100 was achieved during a bioaugmented and biostimulated treatment to remove hydrocarbons from aged contaminated soil from oil well drilling operations. Rhamnolipid production and contaminant removal were determined for several treatments of irradiated and non-irradiated soils: nutrient addition (nitrogen and phosphorus), P. putida addition, and addition of both (P. putida and nutrients). The results were compared against a control treatment that consisted of adding only sterilized water to the soils. In treatment with native microorganisms (non-irradiated soils) supplemented with P. putida, the removal of total petroleum hydrocarbons (TPH) was 40.6%, the rhamnolipid production was 1.54 mg/kg, and a surface tension of 64 mN/m was observed as well as a negative correlation (R = -0.54; p < 0.019) between TPH concentration (mg/kg) and surface tension (mN/m), When both bacteria and nutrients were involved, TPH levels were lowered to 33.7%, and biosurfactant production and surface tension were 2.03 mg/kg and 67.3 mN/m, respectively. In irradiated soil treated with P. putida, TPH removal was 24.5% with rhamnolipid generation of 1.79 mg/kg and 65.6 mN/m of surface tension, and a correlation between bacterial growth and biosurfactant production (R = -0.64; p < 0.009) was observed. When the nutrients and P. putida were added, TPH removal was 61.1%, 1.85 mg/kg of biosurfactants were produced, and the surface tension was 55.6 mN/m. In summary, in irradiated and non-irradiated soils, in situ rhamnolipid production by P. putida enhanced TPH decontamination of the soil.
Assuntos
Glicolipídeos/metabolismo , Hidrocarbonetos/metabolismo , Petróleo/metabolismo , Pseudomonas putida/metabolismo , Microbiologia do Solo , Poluentes do Solo/metabolismo , Tensoativos/metabolismo , Pseudomonas putida/crescimento & desenvolvimentoRESUMO
High copper concentration is toxic for living organisms including humans. Biosorption is a bioremediation technique that can remove copper and other pollutants from aqueous medium and soils, consequently cleaning the environment. The aim of this study was, therefore, to investigate the influence of different copper compounds (Cu(II) as CuCl2; Cu(II) as CuSO4; and Cu(I) as CuCl) on copper bioreduction and biosorption using four copper-resistant bacteria isolated from the rhizosphere of two plants (Avena sativa and Plantago lanceolata) in aqueous matrix. Copper resistance profile, bioreduction, and biosorption after 48 h of incubation were evaluated. The isolates displayed high copper resistance. However, isolate A1 did not grow very well in the CuCl2 and isolate T5 was less resistant to copper in aqueous solutions amended with CuCl (Cu(I)). The best copper source for copper bioreduction and biosorption was CuSO4 and the isolates removed as much as ten times more copper than in aqueous solutions amended with the other copper compounds. Moreover, Cu(I) did not succumb to biosorption, although the microbes were resistant to aqueous solutions of CuCl. In summary, Cu(II) from CuSO4 was furthermost susceptible to bioreduction and biosorption for all isolates. This is an indication that copper contamination of the environment from the use of CuSO4 as an agrochemical is amenable to bioremediation.
Assuntos
Sulfato de Cobre/isolamento & purificação , Cobre/isolamento & purificação , Poluentes Ambientais/isolamento & purificação , Acinetobacter/genética , Acinetobacter/crescimento & desenvolvimento , Acinetobacter/isolamento & purificação , Adsorção , Avena/microbiologia , Biodegradação Ambiental , Biomassa , Farmacorresistência Bacteriana , Oxirredução , Raízes de Plantas/microbiologia , Plantago/microbiologia , Pseudomonas putida/genética , Pseudomonas putida/crescimento & desenvolvimento , Pseudomonas putida/isolamento & purificação , RNA Ribossômico 16S/genética , Rizosfera , Soluções , Stenotrophomonas maltophilia/genética , Stenotrophomonas maltophilia/crescimento & desenvolvimento , Stenotrophomonas maltophilia/isolamento & purificação , ÁguaRESUMO
In situ biosurfactant (rhamnolipid) production by Pseudomonas putida CB-100 was achieved during a bioaugmented and biostimulated treatment to remove hydrocarbons from aged contaminated soil from oil well drilling operations. Rhamnolipid production and contaminant removal were determined for several treatments of irradiated and non-irradiated soils: nutrient addition (nitrogen and phosphorus), P. putida addition, and addition of both (P. putida and nutrients). The results were compared against a control treatment that consisted of adding only sterilized water to the soils. In treatment with native microorganisms (non-irradiated soils) supplemented with P. putida, the removal of total petroleum hydrocarbons (TPH) was 40.6%, the rhamnolipid production was 1.54 mg/kg, and a surface tension of 64 mN/m was observed as well as a negative correlation (R = -0.54; p < 0.019) between TPH concentration (mg/kg) and surface tension (mN/m), When both bacteria and nutrients were involved, TPH levels were lowered to 33.7%, and biosurfactant production and surface tension were 2.03 mg/kg and 67.3 mN/m, respectively. In irradiated soil treated with P. putida, TPH removal was 24.5% with rhamnolipid generation of 1.79 mg/kg and 65.6 mN/m of surface tension, and a correlation between bacterial growth and biosurfactant production (R = -0.64; p < 0.009) was observed. When the nutrients and P. putida were added, TPH removal was 61.1%, 1.85 mg/kg of biosurfactants were produced, and the surface tension was 55.6 mN/m. In summary, in irradiated and non-irradiated soils, in situ rhamnolipid production by P. putida enhanced TPH decontamination of the soil.
Assuntos
Glicolipídeos/metabolismo , Hidrocarbonetos/metabolismo , Petróleo/metabolismo , Pseudomonas putida/metabolismo , Microbiologia do Solo , Poluentes do Solo/metabolismo , Tensoativos/metabolismo , Pseudomonas putida/crescimento & desenvolvimentoRESUMO
The present study examined the aerobic metabolism of trimethylamine in Pseudomonas putida A ATCC 12633 grown on tetradecyltrimethylammonium bromide or trimethylamine. In both conditions, the trimethylamine was used as a nitrogen source and also accumulated in the cell, slowing the bacterial growth. Decreased bacterial growth was counteracted by the addition of AlCl(3). Cell-free extracts prepared from cells grown aerobically on tetradecyltrimethylammonium bromide exhibited trimethylamine monooxygenase activity that produced trimethylamine N-oxide and trimethylamine N-oxide demethylase activity that produced dimethylamine. Cell-free extracts from cells grown on trimethylamine exhibited trimethylamine dehydrogenase activity that produced dimethylamine, which was oxidized to methanal and methylamine by dimethylamine dehydrogenase. These results show that this bacterial strain uses two enzymes to initiate the oxidation of trimethylamine in aerobic conditions. The apparent K(m) for trimethylamine was 0.7 mM for trimethylamine monooxygenase and 4.0 mM for trimethylamine dehydrogenase, but both enzymes maintain similar catalytic efficiency (0.5 and 0.4, respectively). Trimethylamine dehydrogenase was inhibited by trimethylamine from 1 mM. Therefore, the accumulation of trimethylamine inside Pseudomonas putida A ATCC 12633 grown on tetradecyltrimethylammonium bromide or trimethylamine may be due to the low catalytic efficiency of trimethylamine monooxygenase and trimethylamine dehydrogenase.
Assuntos
Metilaminas/metabolismo , Oxirredutases N-Desmetilantes/metabolismo , Oxigenases/metabolismo , Pseudomonas putida/metabolismo , Aerobiose , Dimetilaminas/metabolismo , Cinética , Oxirredução , Oxirredutases N-Desmetilantes/antagonistas & inibidores , Pseudomonas putida/crescimento & desenvolvimento , Compostos de Trimetil Amônio/metabolismoRESUMO
AIMS: The aim of this work was to establish if the response to tetradecyltrimethylammonium (TDTMA), a representative quaternary ammonium compound (QAC), involves changes in the phospholipid (PL) composition of Pseudomonas putida A ATCC 12633. METHODS AND RESULTS: Pseudomonas putida was exposed to 50 mg l(-1) of TDTMA for 15 min, and PL composition was analysed. With respect to control values, phosphatidic acid and phosphatidylglycerol increased by 140% and 120%, respectively; cardiolipin decreased about 60%. In TDTMA-adapted bacteria, the most significant change was a 380% increase in phosphatidic acid. Accompanying this change was a 130% increase in phosphatidylglycerol and a 70% decrease in cardiolipin. The changes in adapted cells were reverted after two subcultures without biocide. CONCLUSIONS: Pseudomonas putida responded to TDTMA through quantitative changes in PLs with specific variations in the content of phosphatidic acid, phosphatidylglycerol and cardiolipin. These modifications indicated that these PLs are involved in cellular responses to QACs, utilizing phosphatidic acid principally to neutralize the high positive charge density given for the ammonium quaternary moiety from TDTMA. SIGNIFICANCE AND IMPACT OF THE STUDY: The changes in PL composition give a new insight about the response inflicted by Ps. putida when these bacteria are exposed to QACs.
Assuntos
Desinfetantes/farmacologia , Fosfolipídeos/metabolismo , Pseudomonas putida/efeitos dos fármacos , Compostos de Trimetil Amônio/farmacologia , Cardiolipinas/metabolismo , Meios de Cultura , Relação Dose-Resposta a Droga , Farmacorresistência Bacteriana , Ácidos Fosfatídicos/metabolismo , Fosfatidilgliceróis/metabolismo , Pseudomonas putida/crescimento & desenvolvimento , Pseudomonas putida/metabolismoRESUMO
Cupriavidus necator (formerly Ralstonia eutropha) JMP134, harbouring the catabolic plasmid pJP4, is the best-studied 2,4-dichlorophenoxyacetic acid (2,4-D) herbicide degrading bacterium. A study of the survival and catabolic performance of strain JMP134 in agricultural soil microcosms exposed to high levels of 2,4-D was carried out. When C. necator JMP134 was introduced into soil microcosms, the rate of 2,4-D removal increased only slightly. This correlated with the poor survival of the strain, as judged by 16S rRNA gene terminal restriction fragment length polymorphism (T-RFLP) profiles, and the semi-quantitative detection of the pJP4-borne tfdA gene sequence, encoding the first step in 2,4-D degradation. After 3 days of incubation in irradiated soil microcosms, the survival of strain JMP134 dramatically improved and the herbicide was completely removed. The introduction of strain JMP134 into native soil microcosms did not produce detectable changes in the structure of the bacterial community, as judged by 16S rRNA gene T-RFLP profiles, but provoked a transient increase of signals putatively corresponding to protozoa, as indicated by 18S rRNA gene T-RFLP profiling. Accordingly, a ciliate able to feed on C. necator JMP134 could be isolated after soil enrichment. In native soil microcosms, C. necator JMP134 survived better than Escherichia coli DH5alpha (pJP4) and similarly to Pseudomonas putida KT2442 (pJP4), indicating that species specific factors control the survival of strains harbouring pJP4. The addition of cycloheximide to soil microcosms strongly improved survival of these three strains, indicating that the eukaryotic microbiota has a strong negative effect in bioaugmentation with catabolic bacteria.
Assuntos
Ácido 2,4-Diclorofenoxiacético/metabolismo , Cupriavidus necator/metabolismo , Eucariotos/metabolismo , Viabilidade Microbiana , Microbiologia do Solo , Animais , Biodegradação Ambiental , Biodiversidade , Cilióforos/isolamento & purificação , Cupriavidus necator/genética , Cupriavidus necator/crescimento & desenvolvimento , Impressões Digitais de DNA , DNA Bacteriano/análise , DNA Bacteriano/genética , DNA de Protozoário/análise , DNA de Protozoário/genética , DNA Ribossômico/análise , DNA Ribossômico/genética , Escherichia coli/crescimento & desenvolvimento , Eucariotos/genética , Eucariotos/isolamento & purificação , Plasmídeos/genética , Polimorfismo de Fragmento de Restrição , Pseudomonas putida/crescimento & desenvolvimento , RNA Ribossômico 16S/genética , RNA Ribossômico 18S/genéticaRESUMO
Las Catonas stream (Buenos Aires Metropolitan Area) receives a complex mixture of pollutants from point and diffuse sources because of the agricultural, industrial and urban land uses of its basin. Widespread detection of heavy metals exceeding aquatic life protection levels has occurred in monitoring reconnaissance studies in surface and pore water. As a result of the screening of Cu, Cd, Zn and Pb resistant/tolerant and culturable microbiota, B101N and 200H strains (Pseudomonas fluorescens or putida) were isolated and selected for further studies. They showed 65% Cd and 35% Zn extraction efficiency from aqueous phase. The potential use of these strains in wastewater treatment is currently investigated in order to contribute to decrease heavy metal pollution, a problem affecting every stream of Buenos Aires Metropolitan Area.
Assuntos
Metais Pesados/metabolismo , Pseudomonas fluorescens/metabolismo , Pseudomonas putida/metabolismo , Microbiologia da Água , Poluentes Químicos da Água/metabolismo , Argentina , Biodegradação Ambiental , Cidades , Monitoramento Ambiental , Metais Pesados/análise , Pseudomonas fluorescens/crescimento & desenvolvimento , Pseudomonas fluorescens/isolamento & purificação , Pseudomonas putida/crescimento & desenvolvimento , Pseudomonas putida/isolamento & purificação , Rios/química , Rios/microbiologia , Poluentes Químicos da Água/análiseRESUMO
The ability of Azospirillum brasilense Cd to colonize the niche occupied by 3 bacterial strains previously isolated from sorghum rhizosphere was studied by means of the Biolog system. The isolates were identified by different methods as strains belonging to Pseudomonas putida, Stenotrophomonas maltophilia, and Klebsiella terrigena species. Several C sources, also chosen among the constituents of sorghum root exudates, were used to evaluate the metabolic profiles of Azospirillum and the sorghum rhizobacteria. Azospirillum brasilense Cd exploited the same class of C compounds as the sorghum rhizobacteria and overlapped in their niche requirements. Since structure and functioning of a microbial community are largely affected by the flow rate of nutrient supply, the competitive behavior of A. brasilense Cd was studied in a chemostat mixed culture under C-limited conditions using disodium succinate as C source. Only at high growth rates, i.e., when the C source was highly supplied, A. brasilense Cd appeared to be a good competitor and it became the dominant species, whereas at low growth rates, it was outnumbered by the other species. However, the coexistence of all the strains was always maintained, thus suggesting that interactions other than competition or a potential cross-feeding might occur within the mixed culture.
Assuntos
Azospirillum brasilense/crescimento & desenvolvimento , Klebsiella/crescimento & desenvolvimento , Pseudomonas putida/crescimento & desenvolvimento , Sorghum/microbiologia , Stenotrophomonas maltophilia/crescimento & desenvolvimento , Aminoácidos/isolamento & purificação , Aminoácidos/metabolismo , Azospirillum brasilense/metabolismo , Biomassa , Metabolismo dos Carboidratos , Carboidratos/isolamento & purificação , Ácidos Carboxílicos/isolamento & purificação , Ácidos Carboxílicos/metabolismo , Técnicas de Cocultura , Contagem de Colônia Microbiana , Klebsiella/isolamento & purificação , Extratos Vegetais/química , Extratos Vegetais/metabolismo , Pseudomonas putida/isolamento & purificação , Microbiologia do Solo , Stenotrophomonas maltophilia/isolamento & purificaçãoRESUMO
Pseudomonas putida P111 was isolated by enrichment culture on 2,5-dichlorobenzoate and was also able to grow on 2-chloro-, 3-chloro-, 4-chloro-, 2,3-dichloro-, 2,4-dichloro-, and 2,3,5-trichlorobenzoates. However, 3,5-dichlorobenzoate completely inhibited growth of P111 on all ortho-substituted benzoates that were tested. When 3,5-dichlorobenzoate was added as a cosubstrate with either 3- or 4-chlorobenzoate, cell yields and chloride release were greater than those observed from growth on either monochlorobenzoate alone. Moreover, resting cells of P111 grown on 4-chlorobenzoate released chloride from 3,5-dichlorobenzoate and produced no identifiable intermediate. In contrast, resting cells grown on 2,5-dichlorobenzoate metabolized 3,5-dichlorobenzoate without release of chloride and accumulated a degradation product, which was identified as 1-carboxy-1,2-dihydroxy-3,5-dichlorocyclohexadiene on the basis of gas chromatography-mass spectrometry confirmation of its two acid-hydrolyzed products, 3,5- and 2,4-dichlorophenol. Since 3,5-dichlorocatechol was rapidly metabolized by cells grown on 2,5-dichlorobenzoate, it is apparent that 1-carboxy-1,2-dihydroxy-3,5-dichlorocyclohexadiene is not further metabolized by these cells. Moreover, induction of a functional dihyrodiol dehydrogenase would not be required for growth of P111 on other ortho-chlorobenzoates since the corresponding chlorodihydrodiols produced from a 1,2-dioxygenase attack would spontaneously decompose to the corresponding catechols. In contrast, growth on 3-chloro-, 4-chloro-, or 3,5-dichlorobenzoate requires a functional dihydrodiol dehydrogenase, yet only the two monochlorobenzoates appear to induce for it.