164 resultados para o-Quinone Methides


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An industrial wastewater treatment plant at Grindsted, Denmark, has suffered from bulking problems for several years caused by filamentous bacteria. Five strains were isolated from the sludge by micromanipulation, Phylogenetic analysis of the 16S rRNA gene sequences showed that the strains formed a monophyletic cluster in the Alphaproteobacteria, and they were phenotypically different from their closest relatives and from all hitherto known filamentous bacteria described (closest relative Brevundimonas vesicularis ATCC 11426(T), 89(.)8% sequence similarity). In pure culture, the cells (1(.)5-2(.)0 mu m) in filaments are Gram-negative and contain polyphosphate and polyhydroxyalkanoates. The optimum temperature for growth is 30 degrees C and the strains grow in 2 % NaCl and are oxidase- and catalase-positive. Ubiquinone 10 is the major quinone. The major fatty acid (C-18: 1 omega 7c) and smaller amounts of unsaturated fatty acids, 3-hydroxy fatty acids with a chain length of 16 and 18 carbon atoms and small amounts of 10-methyl-branched fatty acids with 18 carbon atoms (C-19: 0 10-methyl) affiliated the strains with the Methylobacterium/Xanthobacter group in the Alphaproteobacteria. The G + C content of the DNA is 42(.)9 mol% (for strain Gr1(T)). The two most dissimilar isolates by 16S rRNA gene comparison (Gr1(T) and Gr10; 97(.)7 % identical) showed 71(.)5 % DNA-DNA relatedness. Oligonucleotide probes specific for the pure cultures were designed for fluorescence in situ hybridization and demonstrated that two filamentous morphotypes were present in the Grindsted wastewater treatment plant. It is proposed that the isolates represent a new genus and species, Meganema perideroedes gen. nov., sp. nov. The type strain of Meganema perideroedes is strain Gr1(T) (=DSM 15528(T) =ATCC BAA-740(T)).

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Polymers are subject to oxidation throughout their lifecycle. Antioxidants are generally incorporated in polymers to inhibit or minimise oxidative degradation. Hindered phenolic antioxidants are important stabilisers for polyolefins. However, hindered phenols undergo chemical transformations while performing their antioxidant function during processing and fabrication. In addition, antioxidants are subject to loss from polymers during processing, or subsequently in-service. Migration of antioxidants is a major concern in applications involving polymers in direct contact with food and human environment. This concern is compounded by the realisation that very little is known about the nature and the migration behaviour of antioxidant transformation products. In this work, the antioxidant role of the biological antioxidant -tocopherol (Vitamin E) , which is structurally similar to many synthetic hindered phenols, is investigated in low density polyethylene (LDPE) and polypropylene (PP). The melt stabilising effectiveness of -tocopherol (Toc) was found to be very high, higher than that of commercial hindered phenol antioxidants, such as Irganox 1076 (Irg 1076) and Irganox 1010 (Irg 1010), after multiple extrusions, especially at very low concentrations. The high antioxidant activity of Toc was shown to be due, at least in part, to the formation of transformation products during processing. The main products formed are stereoisomers of dimers and trimers, as well as aldehydes and a quinone - the relative concentration of each was shown to depend on the processing severity, the initial antioxidant concentration and oxygen availability. These transformation products are shown to impart better, similar or lower melt stability to the polymer than the parent antioxidant. The nature of the products formed from Toc during processing was compared with those formed during processing of Irg 1076 and Irg 1010 with LDPE and a mechanism for the melt stabilisation of Toc was proposed and compared with the stabilisation mechanisms of the synthetic antioxidants Irg 1076 and Irg 1010.

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A large number of compounds containing quinonoid or hindered phenol functions were examined for their roles as antifatigue agents. Among the evaluated quinones and phenols expected to have macroalkyl radical scavenging ability, BQ, αTOC, γTOC and GM showed relatively good performance for fatigue resistance (although their performance was slightly less effective than the commercial aromatic amine antioxidants, IPPD and 6PPD). The compounds which were shown to have higher reactivity with alkyl radicals (via calculated reactivity indices) showed better fatigue resistance. This fact supports the suggestion that strong alkyl radical scavengers should be also effective antifatigue agents. Evidence produced based on calculation of reactivity indices suggests that the quinones examined react with alkyl radicals on the meta position of the quinone rings producing phenoxyl radicals. The phenoxyl radicals are expected either to disproportionate, to recombine with a further alkyl radical, or to abstract a hydrogen from another alkyl radical producing an olefine. The regeneration of quinones and formation of the corresponding phenols is expected to occur during the antifatigue activity. The phenol antioxidant, HBA is expected to produce a quinonoid compound and this is also expected to function in a similar way to other quinones. Another phenol, GM, which is also known to scavenge alkyl radicals showed good antifatigue performance. Tocopherols had effective antifatigue activity and are expected to have different antifatigue mechanisms from that of other quinones, hence αTOC was examined for its mechanisms during rubber fatiguing using HPLC analysis. Trimers of αTOC which were produced during vulcanisation are suggested to contribute to the fatigue activity observed. The evidence suggests that the trimers reproduce αTOC and a mechanism was proposed. Although antifatigue agents evaluated showed antifatigue activity, most of them had poor thermoxidative resistance, hence it was necessary to compensate for this by using a combination of antioxidants with the antifatigue agents. Reactive antioxidants which have the potential to graft on the polymer chains during reactive processing were used for this purpose. APMA was the most effective antioxidant among other evaluated reactive antioxidants. Although high ratio of grafting was achieved after optimisation of grafting conditions, it is suggested that this was achieved by long branches of APMA due to large extent of polymerisation. This is expected to cause maldistribution of APMA leading to reducing the effect of CB-D activity (while CB-A activity showed clear advantages for grafting). Further optimisation of grafting conditions is required in order to use APMA more effectively. Moreover, although synergistic effects between APMA and antifatigue agents were expected, none of the evaluated antifatigue agents, BQ, αTOC, γTOC and TMQ, showed significant synergism both in fatigue and thermoxidative resistance. They performed just as additives.

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The effect of substituents on the value of the oxidation potential of quinones is reviewed and attempts to prepare substituted diphenoquinones with high oxidation potentials are reported. Attempts to characterise the mechanism of addition and substitution in diphenoquinones by identifying the products of the Thiele acetylation of diphenoquinone are reported. The reaction proved most efficient when the incoming acetylinium ion is directed by substituents in the diphenoquinone. A 1,8-addition to diphenoquinone is reported and characterised by isolating the products of the reaction between acetyl chloride and diphenoquinone, with perchloric acid as catalyst. The alternating linewidth effects observed in e.s.r.spectra are discussed and applied to account for such effects observed in the e.s.r.spectra of diphenosemiquinone anion and cation radicals. The spectra are analysed and the intramolecular processes producing these effects are discussed. A dianion diradical where intramolecular rotation about the 1 - 1' bond is restricted is produced by the oxidation of 2,2' ,4,4' -tetra hydroxybiphenyl. Previous studies of diphenosemiquinone anions are reviewed and alkylated diphenosemiquinone anion are produced by the reduction of the parent quinone with potassium hydroxide solution, the resulting radical being stabilised by the presence of pyridine. A qualitative interpretation of the solvent-ion effect in alkylated diphenosemiquinone anions is given. Diphanosemiquinone cation radicals are reviewed and previous studies are re-examined.

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Recreational abuse of the drugs cocaine, methamphetamine, and morphine continues to be prevalent in the United States of America and around the world. While numerous methods of detection exist for each drug, they are generally limited by the lifetime of the parent drug and its metabolites in the body. However, the covalent modification of endogenous proteins by these drugs of abuse may act as biomarkers of exposure and allow for extension of detection windows for these drugs beyond the lifetime of parent molecules or metabolites in the free fraction. Additionally, existence of covalently bound molecules arising from drug ingestion can offer insight into downstream toxicities associated with each of these drugs. This research investigated the metabolism of cocaine, methamphetamine, and morphine in common in vitro assay systems, specifically focusing on the generation of reactive intermediates and metabolites that have the potential to form covalent protein adducts. Results demonstrated the formation of covalent adduction products between biological cysteine thiols and reactive moieties on cocaine and morphine metabolites. Rigorous mass spectrometric analysis in conjunction with in vitro metabolic activation, pharmacogenetic reaction phenotyping, and computational modeling were utilized to characterize structures and mechanisms of formation for each resultant thiol adduction product. For cocaine, data collected demonstrated the formation of adduction products from a reactive arene epoxide intermediate, designating a novel metabolic pathway for cocaine. In the case of morphine, data expanded on known adduct-forming pathways using sensitive and selective analysis techniques, following the known reactive metabolite, morphinone, and a proposed novel metabolite, morphine quinone methide. Data collected in this study describe novel metabolic events for multiple important drugs of abuse, culminating in detection methods and mechanistic descriptors useful to both medical and forensic investigators when examining the toxicology associated with cocaine, methamphetamine, and morphine.

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The uncontrolled disposal of wastewaters containing phenolic compounds by the industry has caused irreversible damage to the environment. Because of this, it is now mandatory to develop new methods to treat these effluents before they are disposed of. One of the most promising and low cost approaches is the degradation of phenolic compounds via photocatalysis. This work, in particular, has as the main goal, the customization of a bench scale photoreactor and the preparation of catalysts via utilization of char originated from the fast pyrolysis of sewage sludge. The experiments were carried out at constant temperature (50°C) under oxygen (410, 515, 650 and 750 ml min-1). The reaction took place in the liquid phase (3.4 liters), where the catalyst concentration was 1g L-1 and the initial concentration of phenol was 500 mg L-1 and the reaction time was set to 3 hours. A 400 W lamp was adapted to the reactor. The flow of oxygen was optimized to 650 ml min-1. The pH of the liquid and the nature of the catalyst (acidified and calcined palygorskite, palygorskite impregnated with 3.8% Fe and the pyrolysis char) were investigated. The catalytic materials were characterized by XRD, XRF, and BET. In the process of photocatalytic degradation of phenol, the results showed that the pH has a significant influence on the phenol conversion, with best results for pH equal to 5.5. The phenol conversion ranged from 51.78% for the char sewage sludge to 58.02% (for palygorskite acidified calcined). Liquid samples analyzed by liquid chromatography and the following compounds were identified: hydroquinone, catechol and maleic acid. A mechanism of the reaction was proposed, whereas the phenol is transformed into the homogeneous phase and the others react on the catalyst surface. For the latter, the Langmuir-Hinshelwood model was applied, whose mass balances led to a system of differential equations and these were solved using numerical methods in order to get estimates for the kinetic and adsorption parameters. The model was adjusted satisfactorily to the experimental results. From the proposed mechanism and the operating conditions used in this study, the most favored step, regardless of the catalyst, was the acid group (originated from quinone compounds), being transformed into CO2 and water, whose rate constant k4 presented value of 0.578 mol L-1 min-1 for acidified calcined palygorskite, 0.472 mol L-1 min-1 for Fe2O3/palygorskite and 1.276 mol L-1 min-1 for the sludge to char, the latter being the best catalyst for mineralization of acid to CO2 and water. The quinones were adsorbed to the acidic sites of the calcined palygorskite and Fe2O3/palygorskite whose adsorption constants were similar (~ 4.45 L mol-1) and higher than that of the sewage sludge char (3.77 L mol-1).

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The uncontrolled disposal of wastewaters containing phenolic compounds by the industry has caused irreversible damage to the environment. Because of this, it is now mandatory to develop new methods to treat these effluents before they are disposed of. One of the most promising and low cost approaches is the degradation of phenolic compounds via photocatalysis. This work, in particular, has as the main goal, the customization of a bench scale photoreactor and the preparation of catalysts via utilization of char originated from the fast pyrolysis of sewage sludge. The experiments were carried out at constant temperature (50°C) under oxygen (410, 515, 650 and 750 ml min-1). The reaction took place in the liquid phase (3.4 liters), where the catalyst concentration was 1g L-1 and the initial concentration of phenol was 500 mg L-1 and the reaction time was set to 3 hours. A 400 W lamp was adapted to the reactor. The flow of oxygen was optimized to 650 ml min-1. The pH of the liquid and the nature of the catalyst (acidified and calcined palygorskite, palygorskite impregnated with 3.8% Fe and the pyrolysis char) were investigated. The catalytic materials were characterized by XRD, XRF, and BET. In the process of photocatalytic degradation of phenol, the results showed that the pH has a significant influence on the phenol conversion, with best results for pH equal to 5.5. The phenol conversion ranged from 51.78% for the char sewage sludge to 58.02% (for palygorskite acidified calcined). Liquid samples analyzed by liquid chromatography and the following compounds were identified: hydroquinone, catechol and maleic acid. A mechanism of the reaction was proposed, whereas the phenol is transformed into the homogeneous phase and the others react on the catalyst surface. For the latter, the Langmuir-Hinshelwood model was applied, whose mass balances led to a system of differential equations and these were solved using numerical methods in order to get estimates for the kinetic and adsorption parameters. The model was adjusted satisfactorily to the experimental results. From the proposed mechanism and the operating conditions used in this study, the most favored step, regardless of the catalyst, was the acid group (originated from quinone compounds), being transformed into CO2 and water, whose rate constant k4 presented value of 0.578 mol L-1 min-1 for acidified calcined palygorskite, 0.472 mol L-1 min-1 for Fe2O3/palygorskite and 1.276 mol L-1 min-1 for the sludge to char, the latter being the best catalyst for mineralization of acid to CO2 and water. The quinones were adsorbed to the acidic sites of the calcined palygorskite and Fe2O3/palygorskite whose adsorption constants were similar (~ 4.45 L mol-1) and higher than that of the sewage sludge char (3.77 L mol-1).

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Thesis (Ph.D.)--University of Washington, 2016-08

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Highly purified, intact chloroplasts were prepared from pea (Pisum sativum L.) and spinach (Spinacia oleracea L.) following an identical procedure, and were used to investigate the cupric cation inhibition on the photosynthetic activity. In both species, copper inhibition showed a similar inhibitor concentration that decreases the enzyme activity by 50% (IC(50) approximately 1.8 microM) and did not depend on the internal or external phosphate (Pi) concentration, indicating that copper did not interact with the Pi translocator. Fluorescence analysis suggested that the presence of copper did not facilitate photoinhibition, because there were no changes in maximal fluorescence (F(m)) nor in basal fluorescence (F(o)) of copper-treated samples. The electron transport through the photosystem II (PSII) was also not affected (operating efficiency of PSII-F'v/F'm similar in all conditions). Yet, under Cu(2+) stress, the proportion of open PSII reaction centers was dramatically decreased, and the first quinone acceptor (Q(A)) reoxidation was fully inhibited, as demonstrated by the constant photochemical quenching (q(P)) along experiment time. The quantum yield of PSII electron transport (Phi(PSII)) was also clearly affected by copper, and therefore reduced the photochemistry efficiency. Manganese, when added simultaneously with copper, delayed the inhibition, as measured by oxygen evolution and chlorophyll fluorescence, but neither reversed the copper effect when added to copper-inhibited plastids, nor prevented the inhibition of the Hill activity of isolated copper-treated thylakoids. Our results suggest that manganese competed with copper to penetrate the chloroplast envelope. This competition seems to be specific because other divalent cations e.g. magnesium and calcium, did not interfere with the copper action in intact chloroplasts. All results do suggest that, under these conditions, the stroma proteins, such as the Calvin-Benson cycle enzymes or others are the most probable first target for the Cu(2+) action, resulting in the total inhibition of chloroplast photosynthesis and in the consequent unbalanced rate of production and consumption of the reducing power.

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La ruta de asimilación de cianuro en P. pseudoalcaligenes CECT5344 transcurre a través de un nitrilo formado por la reacción química del cianuro con el oxalacetato, siendo este último acumulado como consecuencia de la acción conjunta de una malato:quinona oxidoreductasa (MQO) y la oxidasa terminal resistente a cianuro (CioAB) (Luque-Almagro et al., 2011b). Los nitrilos pueden ser convertidos en amonio por la acción de una nitrilasa o un sistema nitrilo hidratasa/amidasa. Con el objetivo de elucidar la ruta de asimilación de cianuro en P. pseudoalcalígenes CECT5344, se ha analizado el proteoma de este microorganismo en condiciones cianotróficas frente a nitrato como fuente de nitrógeno como control. En este estudio se identificaron proteínas relacionadas con la ruta de asimilación de cianuro en la estirpe CECT5344, que aparecían inducidas por cianuro, como NitB y NitG, cuyos genes se encuentran localizados en la agrupación génica nit1C. Además de NitB y NitG, de función desconocida, la agrupación génica nit1C codifica un regulador transcripcional del tipo Fis dependiente de σ54 (NitA), una nitrilasa (NitC), una proteína que pertenece a la superfamilia S-adenosilmetionina (NitD), un miembro de la superfamilia N-aciltransferasa (NitE), un polipéptido de la familia AIRS/GARS (NitF) y una oxidorreductasa dependiente de NADH (NitH). Un análisis transcripcional mediante RT-PCR determinó que los genes nitBCDEFGH se cotranscriben, mientras que el gen regulador nitA se transcribe de forma divergente. Además, resultados obtenidos por RT-PCR confirman que la expresión de los genes nitBCDEFGH está inducida por cianuro y reprimida por amonio. La relación entre el cianuro y el grupo de genes nit1C queda patente por el fenotipo de los mutantes deficientes nitA, nitB y nitC, incapaces de usar complejos cianuro-metálicos o 2-hidroxinitrilos como única fuente de nitrógeno. Todos estos datos indican que la nitrilasa NitC, junto con la proteína NitB, utilizan de forma específica determinados nitrilos alifáticos como sustrato, entre los que se encuentran el formado durante la asimilación de cianuro (Estepa et al., 2012). Además, entre las proteínas inducidas por cianuro se identificaron una dihidropicolinato sintasa (DapA), una fosfoserina transaminasa (SerC) y una proteína de función desconocida (Orf1), las tres codificadas por genes del operón cio, una cianasa (CynS), la proteína S6 de la subunidad ribosomal 30S (RpsF), una superóxido dismutasa (SodB), la ferritina (Dps), una oxidorreductasa (Fpr) y un factor de elongación P (EF-P). Una vez identificadas, estas proteínas se han analizado funcionalmente y se han localizado en el genoma de P. pseudoalcaligenes CECT5344 los genes correspondientes, así como los genes adyacentes. La inducción de estas proteínas en condiciones cianotróficas sugiere que el metabolismo del cianuro incluye, además de la resistencia y asimilación de este tóxico, otros procesos biológicos relacionados con el metabolismo del cianato y de algunos aminoácidos, el estrés oxidativo y la homeostasis de hierro, entre otros. Por otra parte, el conocimiento en profundidad y la interpretación de la secuencia génica de P. pseudoalcaligenes CECT5344, así como el análisis comparativo frente a organismos no cianotrofos ha permitido entender algunos de los mecanismos implicados en la resistencia y asimilación de cianuro, lo que permitiría conducir a la posterior mejora del proceso de biodegradación de cianuro. Además, el estudio del genoma de la estirpe CECT5344 permitirá explorar la capacidad de este organismo para ser utilizado en procesos de biorremediación de residuos cianurados en los que se encuentran metales y otros tóxicos (Luque-Almagro et al., 2013; Wibberg et al., 2014). En este trabajo se muestran y discuten los resultados de la secuenciación del genoma de P. pseudoalcaligenes, así como el estudio del análisis filogenético y evolutivo de la cepa, estableciéndose de esta manera relaciones con otras especies en base a los genomas secuenciados de las mismas, entre las que destaca P. mendocina ymp relacionada con P. pseudoalcaligenes CECT5344. El estudio de las características del genoma de P. pseudoalcaligenes CECT5344 ha sido completado con un análisis comparativo frente a los genomas de otras especies de Pseudomonas, encontrándose así semejanzas y diferencias en cuanto a la distribución génica funcional. Por último, se muestra un análisis del genoma de P. pseudoalcaligenes CECT5344 en relación con los genes implicados probablemente en los procesos de asimilación de cianuro y residuos cianurados, tales como los codificantes de nitrilasas y aquellos implicados en la resistencia a cianuro como los constituyentes del operón cio que codifican la oxidasa terminal insensible a cianuro. Finalmente, se discute la presencia de genes implicados posiblemente en otros procesos con una alto potencial biotecnológico, tales como la producción de bioplásticos y la biodegradación de diversos contaminantes.

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Tese (doutorado)—Universidade de Brasília, Instituto de Química, 2016.

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We show here that a physical activation process that is diffusion-controlled yields an activated carbon whose chemistry – both elemental and functional – varies radially through the particles. For the ∼100 μm particles considered here, diffusion-controlled activation in CO2 at 800 °C saw a halving in the oxygen concentration from the particle periphery to its center. It was also observed that this activation process leads to an increase in keto and quinone groups from the particle periphery towards the center and the inverse for other carbonyls as well as ether and hydroxyl groups, suggesting the two are formed under CO2-poor and -rich environments, respectively. In contrast to these observations, use of physical activation processes where diffusion-control is absent are shown to yield carbons whose chemistry is radially invariant. This suggests that a non-diffusion limited activation processes should be used if the performance of a carbon is dependent on having a specific optimal pore surface chemical composition.

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Recreational abuse of the drugs cocaine, methamphetamine, and morphine continues to be prevalent in the United States of America and around the world. While numerous methods of detection exist for each drug, they are generally limited by the lifetime of the parent drug and its metabolites in the body. However, the covalent modification of endogenous proteins by these drugs of abuse may act as biomarkers of exposure and allow for extension of detection windows for these drugs beyond the lifetime of parent molecules or metabolites in the free fraction. Additionally, existence of covalently bound molecules arising from drug ingestion can offer insight into downstream toxicities associated with each of these drugs. This research investigated the metabolism of cocaine, methamphetamine, and morphine in common in vitro assay systems, specifically focusing on the generation of reactive intermediates and metabolites that have the potential to form covalent protein adducts. Results demonstrated the formation of covalent adduction products between biological cysteine thiols and reactive moieties on cocaine and morphine metabolites. Rigorous mass spectrometric analysis in conjunction with in vitro metabolic activation, pharmacogenetic reaction phenotyping, and computational modeling were utilized to characterize structures and mechanisms of formation for each resultant thiol adduction product. For cocaine, data collected demonstrated the formation of adduction products from a reactive arene epoxide intermediate, designating a novel metabolic pathway for cocaine. In the case of morphine, data expanded on known adduct-forming pathways using sensitive and selective analysis techniques, following the known reactive metabolite, morphinone, and a proposed novel metabolite, morphine quinone methide. Data collected in this study describe novel metabolic events for multiple important drugs of abuse, culminating in detection methods and mechanistic descriptors useful to both medical and forensic investigators when examining the toxicology associated with cocaine, methamphetamine, and morphine.

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Five strains (1126-1H-08(T), 51B-09, 986-08, 1084B-08 and 424-08) were isolated from diseased rainbow trout. Cells were Gram-negative rods, 0.7 µm wide and 3 µm long, non-endospore-forming, catalase and oxidase positive. Colonies were circular, yellow-pigmented, smooth and entire on TGE agar after 72 hours incubation at 25°C. They grew in a temperature range between 15°C to 30°C, but they did not grow at 37°Cor 42°C. Based on 16S rRNA gene sequence analysis, the isolates belonged to the genus Flavobacterium. Strain 1126-1H-08(T) exhibited the highest levels of similarity with Flavobacterium oncorhynchi CECT 7678(T) and Flavobacterium pectinovorum DSM 6368(T) (98.5% and 97.9% sequence similarity, respectively). DNA-DNA hybridization values were 87 to 99% among the five isolates and ranged from 21 to 48% between strain 1126-1H-08(T), selected as a representative isolate, and the type strains of Flavobacterium oncorhynchi CECT 7678(T) and other phylogenetic related Flavobacterium species. The DNA G+C content of strain 1126-1H-08(T) was 33.2 mol%. The predominant respiratory quinone was MK-6 and the major fatty acids were iso-C15∶0 and C15∶0. These data were similar to those reported for Flavobacterium species. Several physiological and biochemical tests differentiated the novel bacterial strains from related Flavobacterium species. Phylogenetic, genetic and phenotypic data indicate that these strains represent a new species of the genus Flavobacterium, for which the name Flavobacterium plurextorum sp. nov. was proposed. The type strain is 1126-1H-08(T) ( = CECT 7844(T) = CCUG 60112(T)).