Brevibacterium rongguiense sp. nov., isolated from freshwater sediment.

A Gram stain-positive, non-spore-forming, non-motile and rod-shaped actinomycete, strain 5221T, was isolated from the sediment of a river collected at Ronggui in the Pearl River Delta, PR China. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the strain formed a distinct lineage within the genus Brevibacterium and had the highest sequence similarity to Brevibacterium pityocampae Tp12T (96.7 %), followed by Brevibacterium daeguense 2C6-41T (96.5 %), Brevibacterium samyangense SST-8T (96.0 %) and Brevibacterium ravenspurgense 20T (95.9 %). The results of chemotaxonomic analyses, including detecting anteiso-C15 : 0, anteiso-C17 : 0, and C16 : 0 as the major cellular fatty acids, diphosphatidylglycerol, phosphatidylglycerol and three phosphoglycolipids as the polar lipids, MK-8(H2) as the major menaquinone, and a DNA G+C content of 72.4 mol%, supported that strain 5221T is a member of the genus Brevibacterium. Furthermore, low sequence similarities of 16S rRNA gene sequences, differences in fatty acid compositions and differential physiological characteristics such as enzyme activity and carbon sources utilization ability distinguished the isolate from its close relatives. Therefore, strain 5221T represents a novel species of the genus Brevibacterium, for which the name Brevibacterium rongguiense sp. nov. is proposed, with the type strain 5221T (=GDMCC 1.1766T=KACC 21700T).

Duganella lactea sp. nov., Duganella guangzhouensis sp. nov., Duganella flavida sp. nov. and Massilia rivuli sp. nov., isolated from a subtropical stream in PR China and proposal to reclassify Duganella ginsengisoli as Massilia ginsengisoli comb. nov.

Five Gram-stain-negative, catalase- and oxidase-positive, rod-shaped and motile strains (FT50WT, FT80WT, FT92WT, FT94W and FT135WT) were isolated from a subtropical stream in PR China. Comparisons based on 16S rRNA gene sequences showed that strains FT50WT, FT94W and FT135WT take strain Duganella sacchari Sac-22T, and strains FT80WT and FT92WT take strain Duganella ginsengisoli DCY83T as their closest neighbour in the phylogenetic trees, respectively. The G+C contents of strains FT50WT, FT80WT, FT92WT, FT94W and FT135WT were 63.3, 62.4, 62.8, 63.8 and 60.8 %, respectively. The reconstructed phylogenomic tree based on concatenated 92 core genes showed that strains FT50WT, FT80WT, FT94W and FT135WT clustered together with species of the genus Duganella, but strains FT92WT and D. ginsengisoli KCTC 42409T were located in the clades of the genus Massilia. The calculated pairwise average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values among strains FT50WT, FT80WT, FT92WT, FT94W, FT135WT and related strains were in the ranges of 75.6-87.8% and 20.3-33.8% except that the values between strains FT50WT and FT94W were 98.7 and 89.2%, respectively. The respiratory quinone of these five strains was Q-8. The major fatty acids were C16 : 1 ω7c, C16 : 0, C18 : 1 ω7c and C12 : 0. The polar lipids included phosphatidylethanolamine, phosphatidylglycerol and one unidentified phospholipid. Considering the distinct phylogenetic relationships of D. ginsengisoli with species of the genus Massilia in the phylogenomic tree, it was reasonable to transfer D. ginsengisoli to the genus Massilia as Massilia ginsengisoli comb. nov. Combining the results of phylogenomic analysis, ANI and dDDH data, and a range of physiological and biochemical characteristics together, strains FT50WT and FT94W should belong to the same species and be assigned to genus Duganella with strains FT80WT and FT135WT together, and strain FT92WT should be assigned to the genus Massilia, for which the names Duganella lactea sp. nov. (type strain FT50WT=GDMCC 1.1674T=KACC 21466T), Duganella guangzhouensis sp. nov. (FT80WT=GDMCC 1.1678T=KACC 21470T), Duganella flavida sp. nov. (FT135WT=GDMCC 1.1745T=KACC 21659T) and Massilia rivuli sp. nov. (FT92WT=GDMCC 1.1682T=KACC 21474T) are proposed.

Duganella alba sp. nov., Duganella aquatilis sp. nov., Duganella margarita sp. nov. and Duganella levis sp. nov., isolated from subtropical streams in China.

Six Gram-stain-negative, catalase- and oxidase-positive, rod-shaped and motile strains (FT9WT, FT25W, FT26WT, FT109WT, FT134W and CY42WT) were isolated from subtropical streams in China. Comparisons based on 16S rRNA gene sequences showed that the six strains shared similarities of less than 98.1 % with other species within the family Oxalobacteraceae and formed two separately distinct clades in phylogenetic trees. The 16S rRNA gene sequence similarities between strains FT9WT and FT25W, and between strains FT109WT and FT134W were both 99.7 %. The genome sizes of strains FT9WT, FT25W, FT26WT, FT109WT, FT134W and CY42WT were 6.45, 6.45, 6.54, 6.43, 6.52 and 6.74 Mbp with G+C contents of 64.0, 64.0, 63.8, 63.2, 63.2 and 62.5 %, respectively. The calculated pairwise average nucleotide (ANI) values among the six strains and other related species were less than 93.9 %, except that the values were 99.9 % between strains FT9WT and FT25W, 98.2 % between strains FT109WT and FT134W, and 95.0 and 95.1 % between strain FT26WT and strains FT9WT and FT25W, respectively. However, strain FT26WT shared 16S rRNA gene sequence similarities of only 98.3 and 98.2 % with FT9WT and FT25W, respectively. The respiratory quinone of the six strains was determined to be Q-8. The major fatty acids were C16 : 1ω7c, C16 : 0 and C12 : 0. The predominant polar lipids included phosphatidylethanolamine and phosphatidylglycerol. Considering the phenotypic, biochemical, genotypic and ANI data, strains FT9WT and FT25W, and FT109WT and FT134W may belong to the same species, respectively. Although the pairwise ANI values between strain FT26WT and each of strains FT9WT and FT25W were located in the transition region of species demarcation, the dissimilarities among them indicated that strain FT26WT could represent an independent novel species. The reconstructed phylogenomic tree based on a concatenation of 92 core genes showed that the six strains clustered closely with Duganella sacchari Sac-22T and Duganella radicis KCTC 22382T, and supported that these six strains belong to the genus Duganella. The names Duganella albus sp. nov. (type strain FT9WT=GDMCC 1.1637T=KACC 21313T), Duganella aquatilis sp. nov. (type strain FT26WT=GDMCC 1.1641T=KACC 21315T), Duganella pernnla sp. nov. (type strain FT109WT=GDMCC 1.1688T=KACC 21480T) and Duganella levis sp. nov. (type strain CY42WT=GDMCC 1.1673T=KACC 21465T) are proposed.

Ciceribacter ferrooxidans sp. nov., a nitrate-reducing Fe(II)-oxidizing bacterium isolated from ferrous ion-rich sediment.

A nitrate-reducing Fe(II)-oxidizing bacterial strain, F8825T, was isolated from the Fe(II)-rich sediment of an urban creek in Pearl River Delta, China. The strain was Gram-negative, facultative chemolithotrophic, facultative anaerobic, non-spore-forming, and rod-shaped with a single flagellum. Phy-logenetic analysis based on 16S rRNA gene sequencing indicated that it belongs to the genus Ciceribacter and is most closely related to C. lividus MSSRFBL1T (99.4%), followed by C. thiooxidans F43bT (98.8%) and C. azotifigens A.slu09T (98.0%). Fatty acid, polar lipid, respiratory quinone, and DNA G + C content analyses supported its classification in the genus Ciceribacter. Multilocus sequence analysis of concatenated 16S rRNA, atpD, glnII, gyrB, recA, and thrC suggested that the isolate was a novel species. DNA-DNA hybridization and genome sequence comparisons (90.88 and 89.86%, for values of ANIm and ANIb between strains F8825T with MSSRFBL1T, respectively) confirmed that strain F8825T was a novel species, different from C. lividus MSSRFBL1T, C. thiooxidans F43bT, and C. azotifigens A.slu09T. The physiological and biochemical properties of the strain, such as carbon source utilization, nitrate reduction, and ferrous ion oxidation, further supported that this is a novel species. Based on the polyphasic taxonomic results, strain F8825T was identified as a novel species in the genus Ciceribacter, for which the name Ciceribacter ferrooxidans sp. nov. is proposed. The type strain is F8825T (= CCTCC AB 2018196T = KCTC 62948T).

Janthinobacterium violaceinigrum sp. nov., Janthinobacterium aquaticum sp. nov. and Janthinobacterium rivuli sp. nov., isolated from a subtropical stream in China.

Four strains assigned the names FT13WT, FT14W, FT58WT and FT68WT were isolated from a subtropical stream in PR China. All the strains were Gram-stain-negative, catalase- and oxidase-positive, rod-shaped and motile with flagella. Comparisons based on 16S rRNA gene sequences showed that strains FT13WT, FT14W, FT58WT and FT68WT belonged to genus Janthinobacterium and shared 16S rRNA gene similarities in the range of 98.8-99.7 % with Janthinobacterium lividum DSM 1522T, Janthinobacterium agaricidamnosum DSM 9628T and 'Janthinobacterium svalbardensis JA-1', respectively. The calculated pairwise average nucleotide identity (ANI) values among the genomes of above seven strains were in the range of 79.0-92.2 %, except that the ANI value was 96.8 % between strain FT13WT and FT14W. The respiratory quinone of strains FT13WT, FT14W, FT58WT and FT68WT was determined to be Q-8. The major fatty acids were C16 : 1 ω7c, C16 : 0, C18 : 1 ω7c and C12 : 0. The polar lipids included phosphatidylethanolamine, phosphatidylglycerol and one unidentified phospholipid. The genome sizes of strains FT13WT, FT14W, FT58WT and FT68WT were 6.45, 6.38, 5.73 and 6.37 Mbp with G+C contents of 63.4, 63.7, 61.6 and 63.1 mol%, respectively. Combining phenotypic, biochemical, genotypic and ANI data, strain FT13WT and FT14W should belong to the same species. The four strains were considered to represent three novel species within genus Janthinobacterium, for which the names Janthinobacterium violaceinigrum sp. nov. (type strain FT13WT=GDMCC 1.1638T=KACC 21319T), Janthinobacterium aquaticum sp. nov. (FT58WT=GDMCC 1.1676T=KACC 21468T) and Janthinobacterium rivuli sp. nov. (FT68WT=GDMCC 1.1677T=KACC 21469T) are proposed.

Functional response of sediment bacterial community to iron-reducing bioaugmentation with Shewanella decolorationis S12.

Bioaugmentation with exogenously functional microbes is a widely used technology in bioengineering and environmental remediation. Generally, the colonization of inoculated bacteria is considered to be the determining factor in technical success. However, increasing reports have shown that bioaugmentation was still effective when the colonization of inoculated bacteria was unsuccessful. Here, an augmentation study with iron-reducing bacteria (IRB, Shewanella decolorationis S12) was conducted in Fe(II)-poor sediments to elucidate the role of exogenously inoculated bacteria for bioaugmentation performance. The results showed that a sufficient amount of IRB inputs enhanced the iron reduction in bioaugmented sediments, even though the exogenous IRB did not colonize after the beginning of the experiment (less than 1% at day 3). The iron reduction function responded to stimulation of the indigenous IRB community such as Clostridium, Cupriavidus, Fervidicella, and Acinetobacter, which comprised less than 1% in the initial sediments. Moreover, compared with microbial community in control sediment, more positive correlations between OTUs were observed for that in S12-added sediments upon network analysis. The pH and oxidation-reduction potential of sediment were found to be the predominant factors shaping the iron-reducing microbial communities. It meant that exogenous IRB successfully trigged functional community via altering microenvironment by the inoculated bacteria. Overall, this study provides a new insight into the understanding of the role of single strain addition in iron-reducing bioaugmentation.

The Role of Enriched Microbial Consortium on Iron-Reducing Bioaugmentation in Sediments.

Microbial iron reduction is an important biogeochemical process and involved in various engineered processes, including the traditional clay dyeing processes. Bioaugmentation with iron reducing bacteria (IRB) is generally considered as an effective method to enhance the activity of iron reduction. However, limited information is available about the role of IRB on bioaugmentation. To reveal the roles of introduced IRB on bioaugmentation, an IRB consortium enriched with ferric citrate was inoculated into three Fe(II)-poor sediments which served as the pigments for Gambiered Guangdong silk dyeing. After bioaugmentation, the dyeabilities of all sediments met the demands of Gambiered Guangdong silk through increasing the concentration of key agent [precipitated Fe(II)] by 35, 27, and 61%, respectively. The microbial community analysis revealed that it was the minor species but not the dominant ones in the IRB consortium that promoted the activity of iron reduction. Meanwhile, some indigenous bacteria with the potential of iron reduction, such as Clostridium, Anaeromyxobacter, Bacillus, Pseudomonas, Geothrix, and Acinetobacter, were also stimulated to form mutualistic interaction with introduced consortium. Interestingly, the same initial IRB consortium led to the different community successions among the three sediments and there was even no common genus increasing or decreasing synchronously among the potential IRB of all bioaugmented sediments. The Mantel and canonical correspondence analysis showed that different physiochemical properties of sediments influenced the microbial community structures. This study not only provides a novel bioremediation method for obtaining usable sediments for dyeing Gambiered Guangdong silk, but also contributes to understanding the microbial response to IRB bioaugmentation.

Removal of emulsified oil from water by fruiting bodies of macro-fungus (Auricularia polytricha).

The aim of this study was to investigate the feasibility of utilizing the fruiting bodies of a jelly macro-fungus Auricularia polytricha as adsorbents to remove emulsified oil from water. The effects of several factors, including temperature, initial pH, agitation speed, and adsorbent dosage, were taken into account. Results showed that the optimized conditions for adsorption of A. polytricha were a temperature of 35°C, pH of 7.5, and agitation speed of 100 rpm. The adsorption kinetics were characterized by the pseudo-first order model, which showed the adsorption to be a fast physical process. The Langmuir-Freundlich isotherm described the adsorption very well and predicted the maximum adsorption capacity of 398 mg g-1, under optimized conditions. As illustrated by scanning electron micrographs, the oil particles were adsorbed onto the hairs covering the bottom surface and could be desorbed by normal temperature volatilization. The material could be used as an emulsified oil adsorbent at least three times, retaining more than 95% of the maximum adsorption capacity. The results demonstrated that the fruiting bodies of A. polytricha can be a useful adsorbent to remove emulsified oil from water.

Utilization of two invasive free-floating aquatic plants (Pistia stratiotes and Eichhornia crassipes) as sorbents for oil removal.

Free-floating aquatic plants Pistia stratiotes and Eichhornia crassipes are well-known invasive species in the tropics and subtropics. The aim of this study was to utilize the plants as cost-effective and environmentally friendly oil sorbents. Multilevel wrinkle structure of P. stratiotes leaf (PL), rough surface of E. crassipes leaf (EL), and box structure of E. crassipes stalk (ES) were observed using the scanning electron microscope. The natural hydrophobic structures and capillary rise tests supported the idea to use P. stratiotes and E. crassipes as oil sorbents. Experiments indicated that the oil sorption by the plants was a fast process. The maximum sorption capacities for different oils reached 5.1-7.6, 3.1-4.8, and 10.6-11.7 g of oil per gram of sorbent for PL, EL, and ES, respectively. In the range of 5-35 °C, the sorption capacities of the plants were not significantly different. These results suggest that the plants can be used as efficient oil sorbents.