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- Chryseobacterium palustre sp. nov. and Chryseobacterium humi sp. nov., isolated from industrially contaminated sedimentsPublication . Pires, Carlos; Carvalho, Maria F.; Marco, Paolo De; Magan, Naresh; Castro, Paula M. L.Two Gram-staining-negative bacterial strains, designated 3A10T and ECP37T, were isolated from sediment samples collected from an industrially contaminated site in northern Portugal. These two organisms were rod-shaped, non-motile, aerobic, catalase- and oxidase-positive and formed yellow colonies. The predominant fatty acids were iso-C15 : 0, anteiso-C15 : 0, iso-C17 : 1{omega}9c and iso-C17 : 0 3-OH. The G+C content of the DNA of strains 3A10T and ECP37T was 43 and 34 mol%, respectively. The major isoprenoid quinone of the two strains was MK-6. 16S rRNA gene sequence analysis revealed that strains 3A10T and ECP37T were members of the family Flavobacteriaceae and were related phylogenetically to the genus Chryseobacterium. Strain 3A10T showed 16S rRNA gene sequence similarity values of 97.2 and 96.6 % to the type strains of Chryseobacterium antarcticum and Chryseobacterium jeonii, respectively; strain ECP37T showed 97.3 % similarity to the type strain of Chryseobacterium marinum. DNA–DNA hybridization experiments revealed levels of genomic relatedness of <70 % between strains 3A10T and ECP37T and between these two strains and the type strains of C. marinum, C. antarcticum and C. jeonii, justifying their classification as representing two novel species of the genus Chryseobacterium. The names proposed for these organisms are Chryseobacterium palustre sp. nov. (type strain 3A10T =LMG 24685T =NBRC 104928T) and Chryseobacterium humi sp. nov. (type strain ECP37T =LMG 24684T =NBRC 104927T).
- Isolation and initial characterization of a bacterial consortium able to mineralize fluorobenzenePublication . Carvalho, M. F.; Alves, C. C. T.; Ferreira, M. I. M.; Marco, P. De; Castro, P. M. L.Fluorinated compounds are known to be more resistant to microbial degradation than other halogenated chemicals. A microbial consortium capable of aerobic biodegradation of fluorobenzene (FB) as the sole source of carbon and energy was isolated by selective enrichment from sediments collected in a drain near an industrial site. A combination of three microbial strains recovered from the enriched consortium was shown to be necessary for complete FB mineralization. Two of the strains (F1 and F3) were classified by 16S rRNA analysis as belonging to the Sphingobacterium/Flavobacterium group, while the third (F4) falls in the -Proteobacteria group, clustering with Alcaligenes species. Strain F4 was consistently found in the liquid cultures in a much greater proportion than strains F1 and F3 (86:8:6 for F4, F1, and F3, respectively). Stoichiometric release of fluoride ions was measured in batch and fed-batch cultures. In batch cultures, the consortium was able to use FB up to concentrations of 400 mg liter 1 and was able to utilize a range of other organic compounds, including 4-fluorophenol and 4-fluorobenzoate. To our knowledge this is the first time biodegradation of FB as a sole carbon source has been reported.
- Isolation and characterization of polymeric galloyl-ester- degrading bacteria from a tannery discharge placePublication . Franco, A. R.; Calheiros, C. S. C.; Pacheco, C. C.; Marco, P. De; Manaia, C. M.The culturable bacteria colonizing the rhizosphere of plants growing in the area of discharge of a tannery effluent were characterized. Relative proportions of aerobic, denitrifying, and sulfate-reducing bacteria were determined in the rhizosphere of Typha latifolia, Canna indica, and Phragmites australis. Aerobic bacteria were observed to be the most abundant group in the rhizosphere, and plant type did not seem to influence the abundance of the bacterial types analyzed. To isolate bacteria able to degrade polyphenols used in the tannery industry, enrichments were conducted under different conditions. Bacterial cultures were enriched with individual polyphenols (tannins Tara, Quebracho, or Mimosa) or with an undefined mixture of tannins present in the tannery effluent as carbon source. Cultures enriched with the effluent or Tara tannin were able to degrade tannic acid. Six bacterial isolates purified from these mixed cultures were able to use tannic acid as a sole carbon source in axenic culture. On the basis of 16S ribosomal DNA sequence analysis, these isolates were closely related to organisms belonging to the taxa Serratia, Stenotrophomonas maltophilia, Klebsiella oxytoca, Herbaspirillum chlorophenolicum, and Pseudomonas putida.
- Enrichment of microbial cultures able to degrade 1,3-dichloro-2 propanol: A comparison between batch and continuous methodsPublication . Bastos, Filipe; Bessa, José; Pacheco, Catarina C.; Marco, Paolo De; Castro, Paula M.L.Microbial cultures able to degrade xenobiotic compounds are the key element for biological treatment of waste effluents and are obtained from enrichment processes. In this study, two common enrichment methods, suspension batch and immobilized continuous, were compared. The main selection factor was the presence of 1,3-dichloro- 2-propanol (1,3-DCP) as the single carbon source. Both methods have successfully enriched microbial consortia able to degrade 1,3-DCP. When tested in batch culture, the degradation rates of 1,3-DCP by the two consortia were different, with the consortia obtained by batch enrichment presenting slightly higher rates. A preliminary morphological and biochemical analysis of the predominant colonial types present in each degrading consortia revealed the presence of different constituting strains. Three bacterial isolates capable of degrading 1,3-DCP as single strains were obtained from the batch enrichments. These strains were classified by 16S rRNA analysis as belonging to the Rhizobiaceae group. Degradation rates of 1,3-DCP were lower when single species were used, reaching 45 mg l−1 d−1, as compared to 74 mg l−1 d−1 of the consortia enriched on the batch method. Mutualistic interactions may explain the better performance of the enriched consortia.
- Isolation and properties of a pure bacterial strain capable of fluorobenzene degradation as sole carbon and energy sourcePublication . Carvalho, M. F.; Jorge, R. Ferreira; Pacheco, C. C.; Marco, P. De; Castro, P. M. L.A pure bacterial strain capable of aerobic biodegradation of fluorobenzene (FB) as the sole carbon and energy source was isolated by selective enrichment from sediments collected from a polluted site. 16S rRNA and fatty acid analyses support that strain F11 belongs to a novel genus within the a-2 subgroup of the Proteobacteria, possibly within a new clade related to the order Rhizobiales. In batch cultures, growth of strain F11 on FB led to stoichiometric release of fluoride ion. Maximum experimental growth rate of 0.04 h-1 was obtained at FB concentration of 0.4 mM. Growth kinetics were described by the Luong model. An inhibitory effect with increasing FB concentrations was observed, with no growth occurring at concentrations higher than 3.9 mM. Strain F11 was shown to be able to use a range of other organic compounds, including other fluorinated compounds such as 2-fluorobenzoate, 4-fluorobenzoate and 4-fluorophenol. To our knowledge, this is the first time biodegradation of FB, as the sole carbon and energy source, by a pure bacterium has been reported.
- Labrys portucalensis sp. nov., a fluorobenzenedegrading bacterium isolated from an industrially contaminated sediment in northern PortugalPublication . Carvalho, Maria F.; Marco, Paolo De; Duque, Anouk F.; Pacheco, Catarina C.; Janssen, Dick B.; Castro, Paula M. L.A detailed classification of a novel bacterial strain, designated F11T, capable of degrading fluorobenzene as a sole carbon and energy source, was performed by using a polyphasic approach. This Gram-negative, rod-shaped, non-motile, non-spore-forming, aerobic bacterium was isolated from a sediment sample collected from an industrially contaminated site in northern Portugal. The predominant whole-cell fatty acids were C19 : 0 cyclo ω8c, C16 : 01 C18 : 1ω7c, C18 : 0,C18 : 0 3-OH and C16 : 0 3-OH. The G+C content of the DNA was 62.9 mol% and the major respiratory quinone was ubiquinone 10 (UQ-10). 16S rRNA gene sequence analysis revealed that strain F11T was a member of the class Alphaproteobacteria and was phylogenetically related to the genus Labrys, having sequence similarities of 95.6 and 93.1% to the type strains of Labrys monachus and Labrys methylaminiphilus, respectively. DNA–DNA hybridization experiments revealed levels of relatedness of ,70% between strain F11T and the type strains of L. monachus and L. methylaminiphilus (38.6 and 34.1 %, respectively), justifying the classification of strain F11T as representing a novel species of the genus Labrys. The name Labrys portucalensis sp. nov. is proposed for this organism. The type strain is F11T (5LMG 23412T5DSM 17916T).
- Aerobic and anoxic growth and nitrate removal capacity of a marine denitrifying bacterium isolated from a recirculation aquaculture systemPublication . Borges, Maria-Teresa; Sousa, André; Marco, Paolo De; Matos, Ana; Hönigova, Petra; Castro, Paula M. L.Bacterial biofilters used in marine recirculation aquaculture systems need improvements to enhance nitrogen removal efficiency. Relatively little is known about biofilter autochthonous population structure and function. The present study was aimed at isolating and characterizing an Autochthonous denitrifying bacterium from a marine biofilter installed at a recirculation aquaculture system. Colonization of four different media in a marine fish farm was followed by isolation of various denitrifying strains and molecular classification of the most promising one, strain T2, as a novel member of the Pseudomonas fluorescens cluster. This strain exhibits high metabolic versatility regarding N and C source utilization and environmental conditions for growth. It removed nitrate through aerobic assimilatory metabolism at a specific rate of 116.2 mg NO3-N g dw _1 h _1. Dissimilatory NO3-N removal was observed under oxic conditions at a limited rate, where transient NO2-N formed represented 22% (0.17 mg L _1) of the maximum transient NO2-N observed under anoxic conditions. Dissimilatory NO3-N removal under anoxic conditions occurred at a specific rate of 53.5 mgNO3-N g dw _1 h _1. The isolated denitrifying strain was able to colonize different materials, such as granular activated carbon (GAC), Filtralite and Bioflow plastic rings, which allow the development of a prototype bioreactor for strain characterization under dynamic conditions and mimicking fish-farm operating conditions.
- Labrys potucalensis, a bacterial strain with the capacity to degrade fluorobenzenePublication . Carvalho, M.F.; Marco, P. de; Duque, A.F.; Pacheco, C.C.; Janssen, D.B.; Castro, Paula M. L.