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Simultaneous heterotrophic nitrification and aerobic denitrification by Chryseobacterium sp. R31 isolated from abattoir wastewater.

Kundu P, Pramanik A, Dasgupta A, Mukherjee S, Mukherjee J - Biomed Res Int (2014)

Bottom Line: From an initial COD value of 583.0 mg/L, 95.54% was removed whilst, from a starting NH4 (+)-N concentration of 55.7 mg/L, 95.87% was removed after 48 h contact.Molecular phylogenetic identification, supported by chemotaxonomic and physiological properties, assigned R31 as a close relative of Chryseobacterium haifense.This is the first report on concomitant carbon oxidation, nitrification, and denitrification in the genus Chryseobacterium and the associated kinetic coefficients.

View Article: PubMed Central - PubMed

Affiliation: Department of Civil Engineering, Jadavpur University, Kolkata 700 032, India.

ABSTRACT
A heterotrophic carbon utilizing microbe (R31) capable of simultaneous nitrification and denitrification (SND) was isolated from wastewater of an Indian slaughterhouse. From an initial COD value of 583.0 mg/L, 95.54% was removed whilst, from a starting NH4 (+)-N concentration of 55.7 mg/L, 95.87% was removed after 48 h contact. The concentrations of the intermediates hydroxylamine, nitrite, and nitrate were low, thus ensuring nitrogen removal. Aerobic denitrification occurring during ammonium removal by R31 was confirmed by utilization of both nitrate and nitrite as nitrogen substrates. Glucose and succinate were superior while acetate and citrate were poor substrates for nitrogen removal. Molecular phylogenetic identification, supported by chemotaxonomic and physiological properties, assigned R31 as a close relative of Chryseobacterium haifense. The NH4 (+)-N utilization rate and growth of strain R31 were found to be higher at C/N = 10 in comparison to those achieved with C/N ratios of 5 and 20. Monod kinetic coefficients, half saturation concentration (K s ), maximum rate of substrate utilization (k), yield coefficient, (Y) and endogenous decay coefficient (K d ) indicated potential application of R31 in large-scale SND process. This is the first report on concomitant carbon oxidation, nitrification, and denitrification in the genus Chryseobacterium and the associated kinetic coefficients.

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Unrooted phylogenetic tree obtained by the neighbor-joining (NJ) method based on 16S rRNA gene sequences depicting the position of strain Chryseobacterium sp. R31 amongst its phylogenetic neighbors. Numbers at nodes designate levels of bootstrap support (%) based on a NJ analysis of 1000 resampled datasets; only values higher than 50% are displayed. Asterisks denote branches that were obtained using the maximum parsimony and maximum likelihood algorithms. NCBI accession numbers are provided in parentheses. Bar = 0.1 nucleotide substitutions per site. The sequence of Bacillus subtilis DSM 10 (AJ276351) was applied as an outgroup.
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fig1: Unrooted phylogenetic tree obtained by the neighbor-joining (NJ) method based on 16S rRNA gene sequences depicting the position of strain Chryseobacterium sp. R31 amongst its phylogenetic neighbors. Numbers at nodes designate levels of bootstrap support (%) based on a NJ analysis of 1000 resampled datasets; only values higher than 50% are displayed. Asterisks denote branches that were obtained using the maximum parsimony and maximum likelihood algorithms. NCBI accession numbers are provided in parentheses. Bar = 0.1 nucleotide substitutions per site. The sequence of Bacillus subtilis DSM 10 (AJ276351) was applied as an outgroup.

Mentions: The physiological characteristics of isolate R31 are listed in Supplementary File 2. Small, circular colonies with entire edges were observed when R31 was grown on nutrient agar plates. The colonies were glossy, raised, and yellowish-golden in color. On the basis of nucleotide homology and phylogenetic analysis, the isolate was shown to be significantly similar to Chryseobacterium haifense. Identity analysis on the EZ taxon server [35] revealed that the 16S rRNA gene sequence had the closest similarity (98.25%) to the gene sequence of the type strain of Chryseobacterium haifense (Supplementary File 3). The phylogenetic position of the isolate (R31) having NCBI Genbank accession number KF751764 is shown in the dendogram (Figure 1). Strain R31 emerged as a distinctive phylogenetic line from the cluster containing the type of strains of Chryseobacterium species shown in Supplementary File 3. This differentiation was corroborated by considerable branch length and a high bootstrap value (95%). The phylogenetic position of strain R31 was further supported by the maximum parsimony (MP) and maximum likelihood (ML) analyses. An analogous delineation of the strain was found applying the MP and ML algorithms, which was again confirmed by high bootstrap values (70% for MP and 98% for ML) and significant branch lengths. The result of the fatty acid methyl ester (FAME) analysis is shown in Supplementary File 4. The predominant cellular fatty acids of isolate R31 were 15:0 iso (40.40%), 15:0 anteiso (19.44%), and 17:0 iso 3-OH (11.13%) which closely matched those of Chryseobacterium haifenseT: 15:0 iso (41.6%), 15:0 anteiso (16.6%), and 17:0 iso 3-OH (10.3%) [36]. In the nitrate reduction test, gas formation was observed in Durham's tube indicating that the isolate R31 had the ability to convert nitrate to nitrite and then to nitrogen gas. The isolate (R31) was found positive for both nitrate reductase and nitrite reductase. Similar conclusions were drawn by Fernández et al. [37].


Simultaneous heterotrophic nitrification and aerobic denitrification by Chryseobacterium sp. R31 isolated from abattoir wastewater.

Kundu P, Pramanik A, Dasgupta A, Mukherjee S, Mukherjee J - Biomed Res Int (2014)

Unrooted phylogenetic tree obtained by the neighbor-joining (NJ) method based on 16S rRNA gene sequences depicting the position of strain Chryseobacterium sp. R31 amongst its phylogenetic neighbors. Numbers at nodes designate levels of bootstrap support (%) based on a NJ analysis of 1000 resampled datasets; only values higher than 50% are displayed. Asterisks denote branches that were obtained using the maximum parsimony and maximum likelihood algorithms. NCBI accession numbers are provided in parentheses. Bar = 0.1 nucleotide substitutions per site. The sequence of Bacillus subtilis DSM 10 (AJ276351) was applied as an outgroup.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4060765&req=5

fig1: Unrooted phylogenetic tree obtained by the neighbor-joining (NJ) method based on 16S rRNA gene sequences depicting the position of strain Chryseobacterium sp. R31 amongst its phylogenetic neighbors. Numbers at nodes designate levels of bootstrap support (%) based on a NJ analysis of 1000 resampled datasets; only values higher than 50% are displayed. Asterisks denote branches that were obtained using the maximum parsimony and maximum likelihood algorithms. NCBI accession numbers are provided in parentheses. Bar = 0.1 nucleotide substitutions per site. The sequence of Bacillus subtilis DSM 10 (AJ276351) was applied as an outgroup.
Mentions: The physiological characteristics of isolate R31 are listed in Supplementary File 2. Small, circular colonies with entire edges were observed when R31 was grown on nutrient agar plates. The colonies were glossy, raised, and yellowish-golden in color. On the basis of nucleotide homology and phylogenetic analysis, the isolate was shown to be significantly similar to Chryseobacterium haifense. Identity analysis on the EZ taxon server [35] revealed that the 16S rRNA gene sequence had the closest similarity (98.25%) to the gene sequence of the type strain of Chryseobacterium haifense (Supplementary File 3). The phylogenetic position of the isolate (R31) having NCBI Genbank accession number KF751764 is shown in the dendogram (Figure 1). Strain R31 emerged as a distinctive phylogenetic line from the cluster containing the type of strains of Chryseobacterium species shown in Supplementary File 3. This differentiation was corroborated by considerable branch length and a high bootstrap value (95%). The phylogenetic position of strain R31 was further supported by the maximum parsimony (MP) and maximum likelihood (ML) analyses. An analogous delineation of the strain was found applying the MP and ML algorithms, which was again confirmed by high bootstrap values (70% for MP and 98% for ML) and significant branch lengths. The result of the fatty acid methyl ester (FAME) analysis is shown in Supplementary File 4. The predominant cellular fatty acids of isolate R31 were 15:0 iso (40.40%), 15:0 anteiso (19.44%), and 17:0 iso 3-OH (11.13%) which closely matched those of Chryseobacterium haifenseT: 15:0 iso (41.6%), 15:0 anteiso (16.6%), and 17:0 iso 3-OH (10.3%) [36]. In the nitrate reduction test, gas formation was observed in Durham's tube indicating that the isolate R31 had the ability to convert nitrate to nitrite and then to nitrogen gas. The isolate (R31) was found positive for both nitrate reductase and nitrite reductase. Similar conclusions were drawn by Fernández et al. [37].

Bottom Line: From an initial COD value of 583.0 mg/L, 95.54% was removed whilst, from a starting NH4 (+)-N concentration of 55.7 mg/L, 95.87% was removed after 48 h contact.Molecular phylogenetic identification, supported by chemotaxonomic and physiological properties, assigned R31 as a close relative of Chryseobacterium haifense.This is the first report on concomitant carbon oxidation, nitrification, and denitrification in the genus Chryseobacterium and the associated kinetic coefficients.

View Article: PubMed Central - PubMed

Affiliation: Department of Civil Engineering, Jadavpur University, Kolkata 700 032, India.

ABSTRACT
A heterotrophic carbon utilizing microbe (R31) capable of simultaneous nitrification and denitrification (SND) was isolated from wastewater of an Indian slaughterhouse. From an initial COD value of 583.0 mg/L, 95.54% was removed whilst, from a starting NH4 (+)-N concentration of 55.7 mg/L, 95.87% was removed after 48 h contact. The concentrations of the intermediates hydroxylamine, nitrite, and nitrate were low, thus ensuring nitrogen removal. Aerobic denitrification occurring during ammonium removal by R31 was confirmed by utilization of both nitrate and nitrite as nitrogen substrates. Glucose and succinate were superior while acetate and citrate were poor substrates for nitrogen removal. Molecular phylogenetic identification, supported by chemotaxonomic and physiological properties, assigned R31 as a close relative of Chryseobacterium haifense. The NH4 (+)-N utilization rate and growth of strain R31 were found to be higher at C/N = 10 in comparison to those achieved with C/N ratios of 5 and 20. Monod kinetic coefficients, half saturation concentration (K s ), maximum rate of substrate utilization (k), yield coefficient, (Y) and endogenous decay coefficient (K d ) indicated potential application of R31 in large-scale SND process. This is the first report on concomitant carbon oxidation, nitrification, and denitrification in the genus Chryseobacterium and the associated kinetic coefficients.

Show MeSH
Related in: MedlinePlus