Pilot plant study on nitrogen and phosphorus removal in marine wastewater by marine sediment with sequencing batch reactor.

Effective biological treatment of marine wastewater is not well-known. Accumulation of nitrogen and phosphorus from land-based effluent is a crucial cause of red-tide in marine systems. The purpose of the study is to reduce nitrogen and phosphorus in marine wastewater with a pilot plant-scale sequencing batch reactor (SBR) system by using marine sediment as eco-friendly and effective biological materials, and elucidate which bacterial strains in sludge from marine sediment influence the performance of SBR. By applying eco-friendly high efficiency marine sludge (eco-HEMS), the treatment performance was 15 m3 d-1 of treatment amount in 4.5 m3 of the reactor with the average removal efficiency of 89.3% for total nitrogen and 94.9% for total phosphorus at the optimal operation condition in summer. Moreover, the average removal efficiency was 84.0% for total nitrogen and 88.3% for total phosphorus in winter although biological treatment efficiency in winter is generally lower due to bacterial lower activity. These results were revealed by the DNA barcoding analysis of 16s rRNA amplicon sequencing of samples from the sludge in winter. The comparative analysis of the bacterial community composition in sludge at the high efficiency of the system showed the predominant genera Psychromonas (significantly increased to 45.6% relative abundance), Vibrio (13.3%), Gaetbulibacter (5.7%), and Psychroserpens (4.3%) in the 4 week adaptation after adding marine sediment, suggesting that those predominant bacteria influenced the treatment performance in winter.

Characteristics of crude oil-degrading bacteria Gordonia iterans isolated from marine coastal in Taean sediment.

Crude oil is a major pollutant of marine and coastal ecosystems, and it causes environmental problems more seriously. It is believed ultimate and complete degradation is accomplished mainly by microorganisms. In this study, we aim to search out for bacterial strains with high ability in degrading crude oil. From sediments contaminated by the petroleum spilled in 2007, an accident in Taean, South Korea, we isolated thirty-one bacterial strains in total with potential application in crude oil contamination remediation. In terms of removal percentage after 7 days, one of the strains, Co17, showed the highest removal efficiency with 84.2% of crude oil in Bushnell-Haas media. The Co17 strain even exhibited outstanding ability removing crude oil at a high salt concentration. Through the whole genome sequencing annotation results, many genes related with n-alkane degradation in the genome of Gordonia sp. Co17, revealed alkane-1-monooxygenase, alcohol dehydrogenase, and Baeyer-Villiger monooxygenase. Specially, for confirmation of gene-level, alkB gene encoding alkane hydroxylase (alkane-1-monooxygenase) was found in the strain Co17. The expression of alkB upregulated 125-fold after 18 hr accompany with the removal of n-alkanes of 48.9%. We therefore propose the strain Gordonia iterans Co17, isolated from crude oil-contaminated marine sediment, could be used to offer a new strategy for bioremediation with high efficiency.

Roseomonas deserti sp. nov., isolated from crude oil contaminated desert sand.

Two dark pink pigmented bacterial strains (M3T and M11) were isolated from crude oil contaminated desert sand from Kuwait. Both strains were Gram-stain-negative and small-rod to oval-shaped bacteria. Strains M3T and M11 grew at 13-42 °C (optimum, 30-35 °C) and pH 6.5-9.0 (optimum, 7.0-7.5). No additional NaCl was required for the growth of both strains. The genomic DNA G+C content of strains M3T and M11 were 69.5 and 69.0 mol%, respectively. Both strains were closely related and the mean DNA-DNA hybridization value was 92±1 %. 16S rRNA gene sequence comparisons of both strains indicated that they belong to the genus Roseomonas. Strains M3T and M11 had a sequence similarity of 97.3 and 97.4 % with Roseomonas oryzae JC288T, respectively. Both strains had <97 % 16S rRNA gene sequence similarity with other members of the genus Roseomonas. Strain M3T showed 18±2 and 13±2 % reassociation (based on DNA-DNA hybridization) with R. oryzae KCTC 42542T and Roseomonas cervicalis KACC 11686T, respectively. The major cellular fatty acids (>5 %) were identified as C18 : 1ω6c/C18 : 1ω7c, C16 : 1ω6c/C16 : 1ω7c and C16 : 0 in both strains. Both strains showed diphosphatidylglycerol, phosphatidylglycerol, phosphatidyl-ethanolamine, phosphatidylcholine and unidentified glycolipid as major polar lipids. Based on distinct phenotypic, genotypic and phylogenetic differences from the previously described taxa, we propose the classification of strains M3T and M11 as representative of a novel species in the genus Roseomonas, for which the name Roseomonas deserti sp. nov. is suggested. The type strain is M3T (=KEMB 2255-459T=JCM 31275T).

Nitrogen and phosphorus treatment of marine wastewater by a laboratory-scale sequencing batch reactor with eco-friendly marine high-efficiency sediment.

We screened and identified a NH3-N-removing bacterial strain, Bacillus sp. KGN1, and a [Formula: see text] removing strain, Vibrio sp. KGP1, from 960 indigenous marine isolates from seawater and marine sediment from Tongyeong, South Korea. We developed eco-friendly high-efficiency marine sludge (eco-HEMS), and inoculated these marine bacterial strains into the marine sediment. A laboratory-scale sequencing batch reactor (SBR) system using the eco-HEMS for marine wastewater from land-based fish farms improved the treatment performance as indicated by 88.2% removal efficiency (RE) of total nitrogen (initial: 5.6 mg/L) and 90.6% RE of total phosphorus (initial: 1.2 mg/L) under the optimal operation conditions (food and microorganism (F/M) ratio, 0.35 g SCODCr/g mixed liquor volatile suspended solids (MLVSS)·d; dissolved oxygen (DO) 1.0 ± 0.2 mg/L; hydraulic retention time (HRT), 6.6 h; solids retention time (SRT), 12 d). The following kinetic parameters were obtained: cell yield (Y), 0.29 g MLVSS/g SCODCr; specific growth rate (µ), 0.06 d-1; specific nitrification rate (SNR), 0.49 mg NH3-N/g MLVSS·h; specific denitrification rate (SDNR), 0.005 mg [Formula: see text]/g MLVSS·h; specific phosphorus uptake rate (SPUR), 0.12 mg [Formula: see text]/g MLVSS·h. The nitrogen- and phosphorus-removing bacterial strains comprised 18.4% of distribution rate in the microbial community of eco-HEMS under the optimal operation conditions. Therefore, eco-HEMS effectively removed nitrogen and phosphorus from highly saline marine wastewater from land-based fish farms with improving SNR, SDNR, and SPUR values in more diverse microbial communities. ABBREVIATIONS: DO: dissolved oxygen; Eco-HEMS: eco-friendly high efficiency marine sludge; F/M: food and microorganism ratio; HRT: hydraulic retention time; ML(V)SS: mixed liquor (volatile) suspended solids; NCBI: National Center for Biotechnology Information; ND: not determined; qPCR: quantitative real-time polymerase chain reaction; RE: removal efficiency; SBR: sequencing batch reactor; SD: standard deviation; SDNR: specific denitrification rate; SNR: specific nitrification rate; SPUR: specific phosphate uptake rate; SRT: solids retention time; T-N: total nitrogen; T-P: total phosphorus; (V)SS: (volatile) suspended solids; w.w.: wet weight.

Skermanella mucosa sp. nov., isolated from crude oil contaminated soil.

A novel Gram-stain negative, small rod-shaped bacterium (strain 8-14-6T) was isolated from hydrocarbon contaminated desert soil collected from Kuwait. Strain 8-14-6T grew at 5-37 °C, pH 6.0-8.8 and 0-2% (w/v) of NaCl concentration. Casein, starch, Tween 20 and Tween 80 were hydrolyzed while urea, chitin, DNA and carboxymethyl-cellulose were not hydrolyzed by strain 8-14-6T. The major cellular fatty acids were identified as C18:1ω6c/C18:1ω7c, C16:0 and iso-C16:1I/C14:03-OH. Strain 8-14-6T produced diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, an unidentified phospholipids, two unidentified lipids and five unidentified amino lipids as polar lipids. Genomic G+C content was 73.5 mol %. 16S rRNA gene sequence comparisons indicated that strain 8-14-6T represents a member of the genus Skermanella within family Rhodospirillaceae of the class Alphaproteobacteria. Strain 8-14-6T has a sequence similarity of 98.9% with Skermanella rosea M1T, 97.4% with Skermanella aerolata 5416T-32T, 96.9% with Skermanella stibiiresistens SB22T and <95.4% with the other two known species of the genus Skermanella. The DNA-DNA relatedness values between strain 8-14-6T and the type strains of the closely related species were clearly below the 70% threshold. From the combination of phenotypic and genotypic characteristics and distinct phylogenetic position, the strain is considered to represent a novel species of the genus Skermanella, for which the name Skermanella mucosa sp. nov. is proposed. The type strain is 8-14-6T (=KEMB 2255-438T =JCM 31590T).

Microbacterium rhizosphaerae sp. nov., isolated from a Ginseng field, South Korea.

A novel Gram-stain positive, aerobic, short rod-shaped, non-motile bacterium, designated strain CHO1T, was isolated from rhizosphere soil from a ginseng agriculture field. Strain CHO1T was observed to form yellow colonies on R2A agar medium. The cell wall peptidoglycan was found to contain alanine, glycine, glutamic acid, D-ornithine and serine. The cell wall sugars were identified as galactose, mannose, rhamnose and ribose. Strain CHO1T was found to contain MK-11, MK-12, MK-13 as the predominant menaquinones and anteiso-C15:0, iso-C16:0, and anteiso-C17:0 as the major fatty acids. Diphosphatidylglycerol, phosphatidylglycerol, phosphoglycolipid, an unidentified phospholipid and three unidentified glycolipids were found to be present in strain CHO1T. Based on 16S rRNA gene sequence analysis, strain CHO1T was found to be closely related to Microbacterium mangrovi DSM 28240T (97.81 % similarity), Microbacterium immunditiarum JCM 14034T (97.45 %), Microbacterium oryzae JCM 16837T (97.33 %) and Microbacterium ulmi KCTC 19363T (97.10 %) and to other species of the genus Microbacterium. The DNA G+C content of CHO1T was determined to be 70.1 mol %. The DNA-DNA hybridization values of CHO1T with M. mangrovi DSM 28240T, M. immunditiarum JCM 14034T, M. oryzae JCM 16837T and M. ulmi KCTC 19363T were 46.7 ± 2, 32.4 ± 2, 32.0 ± 2 and 29.2 ± 2 %, respectively. On the basis of genotypic, phenotypic and phylogenetic properties, it is concluded that strain CHO1T represents a novel species within the genus Microbacterium, for which the name Microbacterium rhizosphaerae sp. nov. is proposed. The type strain of M. rhizosphaerae is CHO1T (= KEMB 7306-513T = JCM 31396T).

Oceanisphaera aquimarina sp. nov., Isolated from Oil-Contaminated Sediment of Ocean Coastal Area from South Korea.

Strain S33(T) was isolated from oil-contaminated sediment of Tae-an coastal region of South Korea. Cells are aerobic, motile, Gram staining-negative, and coccoid shaped. Strain S33(T) grew optimally at the temperature of 25 °C (range of 4-40 °C), pH 6.0 (range of pH 6.0-10.0), and in the presence of 1 % (w/v) NaCl (range of 0-10 %). Ubiquinone-8 was the predominant respiratory quinone. C16:0, summed feature 3 (comprising C16:1ω7c/C16:1ω6c) and C18:1ω7c were the major fatty acids. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol. Strain S33(T) showed the ability to degrade benzene, toluene, and ethylbenzene after 3 days incubation. 16S rRNA gene sequence analysis showed that the strain S33(T) was most closely related to Oceanisphaera sediminis TW92(T) (97.3 %), Oceanisphaera profunda SM1222(T) (97.2 %), and Oceanisphaera ostreae T-w6(T) (97.1 %) and <97 % with other members of the genus Oceanisphaera. The genomic DNA G+C mol% content of strain S33(T) was 51.0 mol%. Based on distinct phenotypic, genotypic, and phylogenetic analysis, strain S33(T) was proposed to represent a novel species in the genus Oceanisphaera as Oceanisphaera aquimarina sp. nov. (= KEMB 1002-058(T) = JCM 30 794(T)).

Leucobacter margaritiformis sp. nov., isolated from bamboo extract.

A Gram-positive aerobic rod-shaped non-motile bacterium designated A23(T) was isolated from bamboo extract that had been used to remove odor and was characterized to determine its taxonomic position. 16S rRNA gene sequence analysis revealed that strain A23(T) belongs to the phylum Actinobacteria. The highest degree of sequence similarities was determined to be with Leucobacter salsicius M1-8(T) (96.7%), Leucobacter exalbidus K-540B(T) (96.4%), Leucobacter chromiireducens subsp. chromiireducens L-1(T) (96.4%), Leucobacter komagatae IFO 15245(T) (96.4%) and Leucobacter aerolatus Sj10(T) (96.4%). Chemotaxonomic data revealed that strain A23(T) possesses menaquinone MK11, and its cell wall peptidoglycan contained 2,4-diaminobutyric acid, alanine, glycine, glutamic acid and γ-aminobutyric acid. The polar lipid profile of strain A23(T) contained diphosphatidylglycerol, phosphatidylglycerol and an unknown glycolipid. The predominant fatty acids were iso-C(16:0) (31.5%), anteiso-C(15:0) (43.2%) and anteiso-C(17:0) (13.9%), all of which corroborated the assignment of the strain to the genus Leucobacter. Based on these data, A23(T) (=KEMC 551-022(T) = JCM 17538(T)) should be classified as the type strain for a novel Leucobacter species, for which the name Leucobacter margaritiformis sp. nov. is proposed.