Field application of bio-foam spray to reduce ammonia emission from ammonia-rich swine manure piles.

In this study, the first field-scale application of a bio-foam spray (a mixture of microbes and a surfactant) for the reduction of ammonia emitted from manure was investigated on six field swine manure piles. The objective of this study was to evaluate the odor suppression ability of bio-foam and odor degradation ability of odor-degrading bacteria loaded in the surfactant foam after covering manure piles. The size of field manure piles tested in this study ranged from 27 to 300 m3. Bio-foam spraying completely suppressed the release of the major odor component, ammonia (NH3), and odor-degrading bacteria in the bio-foam aided in the degradation of NH3 in field swine manure piles. On average, 85.7-100% of NH3 was reduced after 24-48 h of serial bio-foam spray application on the swine manure surface, while the control showed 25-42%. The reduction efficiency of NH3 by the bio-foam application was affected by the bio-foam spray frequency, ambient temperature, ventilation of the field facility, and upward airflow to the pile. The reduction in surface emission of NH3 also reduced the ambient air concentration of NH3 at the gate of the compost facility. NH3 gas measurements at a depth of 50 cm indicated that NH3-degrading bacteria infiltrated the manure and were active in biodegradation. Finally, the measured effectiveness of bio-foam application as shown by this study indicates that sprinkling bio-foam via specialized rotating sprinklers may be an efficient and uniform method for the delivery of bio-foam to wide field areas within composting facilities.

Rice Husk-Cellulose-Based Agricultural Waste Enhances the Degradation of Synthetic Dyes Using Multiple Enzyme-Producing Extremophiles.

The brightly colored synthetic dyes used in the textile industry are discharged at high concentrations-for example, various azo dyes including Methylene Blue (MB) and Methyl Orange (MO)-which is a matter of global concern, as such dyes are harmful to humans and the environment. Microbial degradation is considered an efficient alternative for overcoming the disadvantages of conventional physical and chemical dye removal methods. In this study, we investigated the potential of multiple types of the enzyme-producing extremophilic bacteria Bacillus FW2, isolated from food waste leachate, for the decolorization and bioremediation of artificial synthetic dyes. The screening of enzyme production and assaying of bacterial strain enzymes are essential for enhancing the breakdown of azo bonds in textile azo dyes. The degradation efficiencies of the water-soluble dyes MB and MO were determined at different concentrations using rice husk, which is an efficient substrate. Using the rice husks, the MO was removed completely within 20 h, and an estimated 99.8% of MB was degraded after 24 h by employing shaking at 120 rpm at 40 °C-whereas a removal efficiency of 98.9% was achieved for the combination of MB + MO. These results indicate the possibility of applying an extremophilic bacterial strain, Bacillus sp., for large-scale dye degradation in the future.

Chitinophaga nivalis sp. nov., isolated from forest soil in Pyeongchang, Republic of Korea.

Rod-shaped Gram-stain-negative, aerobic bacterial strains, designated PC14 and PC15T, were isolated from a forest soil sample collected in Pyeongchang county, Gangwon-do, Republic of Korea. Strains PC14 and PC15T grew at 15-37 °C (optimum, 28-30 °C in tryptone soya agar and Mueller-Hinton agar), hydrolysed chitin and casein, and tolerated pH 8.5 and 2 % (w/v) NaCl. The strains were most closely related to members of the genus Chitinophaga, namely Chitinophaga arvensicola DSM 3695T (98.4 %), Chitinophaga longshanensis Z29T (98.3 %), Chitinophaga ginsengisegetis Gsoil 040T (97.8 %), Chitinophaga polysaccharea MRP-15T (97.8 %) and Chitinophaga niastensis JS16-4T (97.7 %). The type strain grew well on conventional commercial media in the laboratory, including tryptone soya agar, Mueller-Hinton agar, Reasoner's 2A agar, nutrient agar and Luria-Bertani agar. The major polar lipid profile comprised phosphatidylethanolamine, an unidentified aminolipid and unidentified polar lipids. The major respiratory quinone was menaquinone-7. The main fatty acids were iso-C15:0, C16:1 ω5c, C16:0 3-OH, iso-C15:0 3-OH and iso-C17:0 3-OH. The DNA G+C content of the isolated strain based on the whole genome sequence was 46.6 mol%. The average nucleotide identity and digital DNA-DNA hybridization values between strains PC14 and PC15T and the reference type strains ranged from 71.0 to 76.5 %, and from 20.3 to 20.7 %, respectively. Based on phenotypic, chemotypic and genotypic evidence, strain PC15T could be differentiated phylogenetically and phenotypically from the recognized species of the genus Chitinophaga. Therefore, strain PC15T is considered to represent a novel species, for which the name Chitinophaga nivalis sp. nov. is proposed. The type strain is PC15T (=KACC 22893T=JCM 35788T).

Investigating Bio-Inspired Degradation of Toxic Dyes Using Potential Multi-Enzyme Producing Extremophiles.

Biological treatment methods overcome many of the drawbacks of physicochemical strategies and play a significant role in removing dye contamination for environmental sustainability. Numerous microorganisms have been investigated as promising dye-degrading candidates because of their high metabolic potential. However, few can be applied on a large scale because of the extremely harsh conditions in effluents polluted with multiple dyes, such as alkaline pH, high salinity/heavy metals/dye concentration, high temperature, and oxidative stress. Therefore, extremophilic microorganisms offer enormous opportunities for practical biodegradation processes as they are naturally adapted to multi-stress conditions due to the special structure of their cell wall, capsule, S-layer proteins, extracellular polymer substances (EPS), and siderophores structural and functional properties such as poly-enzymes produced. This review provides scientific information for a broader understanding of general dyes, their toxicity, and their harmful effects. The advantages and disadvantages of physicochemical methods are also highlighted and compared to those of microbial strategies. New techniques and methodologies used in recent studies are briefly summarized and discussed. In particular, this study addresses the key adaptation mechanisms, whole-cell, enzymatic degradation, and non-enzymatic pathways in aerobic, anaerobic, and combination conditions of extremophiles in dye degradation and decolorization. Furthermore, they have special metabolic pathways and protein frameworks that contribute significantly to the complete mineralization and decolorization of the dye when all functions are turned on. The high potential efficiency of microbial degradation by unculturable and multi-enzyme-producing extremophiles remains a question that needs to be answered in practical research.

Pedobacter montanisoli sp. nov., isolated from soil.

A white-pigmented, non-motile, Gram-stain-negative, rod-shaped bacterium, designated CYS-01T, was obtained from soil sampled at Suwon, Gyeonggi-do, Republic of Korea. Cells were strictly aerobic, grew optimally at 28 °C. Phylogenetic analysis based on its 16S rRNA gene sequence revealed that strain CYS-01T formed a lineage within the family Sphingobacteriaceae and clustered with members of the genus Pedobacter. The closest relatives were Pedobacter xixiisoli CGMCC 1.12803T (95.70 % sequence similarity), Pedobacter ureilyticus THG-T11T (95.35 %), Pedobacter helvus P-25T (95.28 %), Pedobacter chitinilyticus CM134L-2T (94.94 %), Pedobacter nanyangensis Q-4T (94.73 %) and Pedobacter zeaxanthinifaciens TDMA-5T (94.07 %). The principal respiratory quinone was MK-7 and the major polar lipids were phosphatidylethanolamine, an unidentified aminolipid, unidentified lipids and an unidentified glycolipid. The predominant cellular fatty acids were iso-C15 : 0, summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c) and iso-C17 : 0 3-OH. The DNA G+C content was 36.6 mol%. Based on the results of genomic, chemotaxonomic, phenotypic and phylogenetic analyses, strain CYS-01T represents novel species in the genus Pedobacter, for which the name Pedobacter montanisoli sp. nov. is proposed. The type strain is CYS-01T (=KACC 22655T=NBRC 115630T).

Antioxidant and Antimicrobial Properties of Fruiting Body and Submerged Mycelium of Medicinal Mushroom Phellinus robiniae (Agaricomycetes).

This study was conducted to evaluate extraction yield, antioxidant content, antioxidant capacity and antibacterial activity of extracts obtained from submerged mycelium (ME) and fruiting body (FBE) of Phellinus robiniae NTH-PR1. The results showed that yields of ME and FBE reached 14.84 ± 0.63 and 18.89 ± 0.86%, respectively. TPSC, TPC, and TFC were present in both mycelium and fruiting body, and the more contents of them were found in fruiting body. The concentrations of TPSC, TPC and TFC in ME and FBE were 17.61 ± 0.67 and 21.56 ± 0.89 mg GE g-1, 9.31 ± 0.45 and 12.14 ± 0.56 mg QAE g-1, and 8.91 ± 0.53 and 9.04 ± 0.74 mg QE g-1, respectively. EC50 values for DPPH radical scavenging revealed FBE (260.62 ± 3.33 µg mL-1) was more effective than ME (298.21 ± 3.61 µg mL-1). EC50 values for ferrous ion chelating in ME and FBE were 411.87 ± 7.27 and 432.39 ± 2.23 µg mL-1, respectively. Thus, both extracts were able to inhibit Gram-positive and Gram-negative pathogenic bacterial strains, at concentrations ranging in 25-100 mg mL-1 of ME and 18.75-75 mg mL-1 of FBE for Gram-positive bacteria; ranging in 75-100 mg mL-1 of ME and 50-75 of FBE for Gram-negative bacteria. Overall submerged mycelial biomass and fruiting bodies of Ph. robiniae NTH-PR1 can be considered as useful natural sources for development of functional food, pharmaceuticals and cosmetic products or cosmeceuticals.

Variovorax terrae sp. nov. Isolated from Soil with Potential Antioxidant Activity.

A white-pigmented, non-motile, gram-negative, and rod-shaped bacterium, designated CYS-02T, was isolated from soil sampled at Suwon, Gyeonggi-do, Republic of Korea. Cells were strictly aerobic, grew optimally at 20-28ºC and hydrolyzed Tween 40. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain CYS-02T formed a lineage within the family Comamonadaceae and clustered as members of the genus Variovorax. The closest members were Variovorax guangxiensis DSM 27352T (98.6% sequence similarity), Variovorax paradoxus NBRC 15149T (98.5%), and Variovorax gossypii JM-310T (98.3%). The principal respiratory quinone was Q-8 and the major polar lipids contain phosphatidylethanolamine (PE), phosphatidylethanolamine (PG), and diphosphatidylglycerol (DPG). The predominant cellular fatty acids were C16:0, summed feature 3 (C16:1ω7c and/or C16:1ω6c) and summed feature 8 (C18:1ω7c and/or C18:1ω6c). The DNA GC content was 67.7 mol%. The ANI and dDDH values between strain CYS-02T and the closest members in the genus Variovorax were ≤ 79.0 and 22.4%, respectively, and the AAI and POCP values between CYS-02T and the other related species in the family Comamonadaceae were > 70% and > 50%, respectively. The genome of strain CYS-02T showed a putative terpene biosynthetic cluster responsible for antioxidant activity which was supported by DPPH radical scavenging activity test. Based on genomic, phenotypic and chemotaxonomic analyses, strain CYS-02T was classified into a novel species in the genus Variovorax, for which the name Variovorax terrae sp. nov., has been proposed. The type strain is CYS-02T (= KACC 22656T = NBRC 115645 [corrected] T).

Bacterial Biosorbents, an Efficient Heavy Metals Green Clean-Up Strategy: Prospects, Challenges, and Opportunities.

Rapid industrialization has led to the pollution of soil and water by various types of contaminants. Heavy metals (HMs) are considered the most reactive toxic contaminants, even at low concentrations, which cause health problems through accumulation in the food chain and water. Remediation using conventional methods, including physical and chemical techniques, is a costly treatment process and generates toxic by-products, which may negatively affect the surrounding environment. Therefore, biosorption has attracted significant research interest in the recent decades. In contrast to existing methods, bacterial biomass offers a potential alternative for recovering toxic/persistent HMs from the environment through different mechanisms for metal ion uptake. This review provides an outlook of the advantages and disadvantages of the current bioremediation technologies and describes bacterial groups, especially extremophiles with biosorbent potential for heavy metal removal with relevant examples and perspectives.

Genome mining revealed polyhydroxybutyrate biosynthesis by Ramlibacter agri sp. nov., isolated from agriculture soil in Korea.

A white-colony-forming, facultative anaerobic, motile and Gram-stain-negative bacterium, designated G-1-2-2 T was isolated from soil of agriculture field near Kyonggi University, Republic of Korea. Strain G-1-2-2 T synthesized the polyhydroxybutyrate and could grow at 10-35 °C. The phylogenetic analysis based on 16S rRNA gene sequence showed that, strain G-1-2-2 T formed a lineage within the family Comamonadaceae and clustered as a member of the genus Ramlibacter. The 16S rRNA gene sequence of strain G-1-2-2 T showed high sequence similarities with Ramlibacter ginsenosidimutans BXN5-27 T (97.9%), Ramlibacter monticola G-3-2 T (97.9%) and Ramlibacter alkalitolerans CJ661T (97.5%). The sole respiratory quinone was ubiquinone-8 (Q-8). The major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, and an unidentified phospholipid. The principal cellular fatty acids were C16:0, cyclo-C17:0, summed feature 3 (C16:1ω7c and/or C16:1ω6c) and summed feature 8 (C18:1ω7c and/or C18:1ω6c). The genome of strain G-1-2-2 T was 7,200,642 bp long with 13 contigs, 6,647 protein-coding genes, and DNA G + C content of 68.9%. The average nucleotide identity and in silico DNA-DNA hybridization values between strain G-1-2-2 T and close members were ≤ 81.2 and 24.1%, respectively. The genome of strain G-1-2-2 T showed eight putative biosynthetic gene clusters responsible for various secondary metabolites. Genome mining revealed the presence of atoB, atoB2, phaS, phbB, phbC, and bhbD genes in the genome which are responsible for polyhydroxybutyrate biosynthesis. Based on these data, strain G-1-2-2 T represents a novel species in the genus Ramlibacter, for which the name Ramlibacter agri sp. nov. is proposed. The type strain is G-1-2-2 T (= KACC 21616 T = NBRC 114389 T).

Coconut Mesocarp-Based Lignocellulosic Waste as a Substrate for Cellulase Production from High Promising Multienzyme-Producing Bacillus amyloliquefaciens FW2 without Pretreatments.

Facing the crucial issue of high cost in cellulase production from commercial celluloses, inexpensive lignocellulosic materials from agricultural wastes have been attractive. Therefore, several studies have focused on increasing the efficiency of cellulase production by potential microorganisms capable of secreting a high and diversified amount of enzymes using agricultural waste as valuable substrates. Especially, extremophilic bacteria play an important role in biorefinery due to their high value catalytic enzymes that are active even under harsh environmental conditions. Therefore, in this study, we aim to investigate the ability to produce cellulase from coconut-mesocarp of the potential bacterial strain FW2 that was isolated from kitchen food waste in South Korea. This strain was tolerant in a wide range of temperature (-6-75 °C, pH range (4.5-12)) and at high salt concentration up to 35% NaCl. The molecular weight of the purified cellulase produced from strain FW2 was estimated to be 55 kDa. Optimal conditions for the enzyme activity using commercial substrates were found to be 40-50 °C, pH 7.0-7.5, and 0-10% NaCl observed in 920 U/mL of CMCase, 1300 U/mL of Avicelase, and 150 U/mL of FPase. It was achieved in 650 U/mL, 720 U/mL, and 140 U/mL of CMCase, Avicelase, and FPase using coconut-mesocarp, respectively. The results revealed that enzyme production by strain FW2 may have significant commercial values for industry, argo-waste treatment, and other potential applications.

Lysobacter terrestris sp. nov., isolated from soil.

A yellow-pigmented, non-motile, Gram-stain-negative, rod-shaped bacterium, designated II4T was obtained from soil sampled at Seongnam, Gyeonggi-do, Republic of Korea. Cells were strictly aerobic, grew optimally at 20-28 °C and hydrolysed casein. A phylogenetic analysis based on its 16S rRNA gene sequence revealed that strain II4T formed a lineage within the family Xanthomonadaceae and clustered as members of the genus Lysobacter. The closest members were Lysobacter terrae THG-A13T (97.88 % sequence similarity), Lysobacter niabensis GH34-4T (97.82 %), Lysobacter oryzae YC6269T (97.74%), Lysobacter yangpyeongensis GH19-3T (97.53 %) and Lysobacter enzymogenes ATCC 29487T (96.18 %). The principal respiratory quinone was Q-8 and the major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The predominant cellular fatty acids were summed feature 9 (C16 : 0 10-methyl and/or iso-C17 : 1 ω9c) and iso-C15 : 0 and iso-C16 : 0. The DNA G+C content was 68.2 mol%. The average nucleotide identity and in silico DNA-DNA hybridization relatedness values between strain II4T and its closely related genus members with possible full genome sequences were ≤79.6 and 23.7 %, respectively. Based on genomic, chemotaxonomic, phenotypic and phylogenetic data, strain II4T represents novel species in the genus Lysobacter, for which the name Lyobacter terrestris sp. nov. is proposed. The type strain is II4T (=KACC 21196T=NBRC 113956T).

Improvement of Hydrogen Production during Anaerobic Fermentation of Food Waste Leachate by Enriched Bacterial Culture Using Biochar as an Additive.

It has become urgent to develop cost-effective and clean technologies for the rapid and efficient treatment of food waste leachate, caused by the rapid accumulation of food waste volume. Moreover, to face the energy crisis, and to avoid dependence on non-renewable energy sources, the investigation of new sustainable and renewable energy sources from organic waste to energy conversion is an attractive option. Green energy biohydrogen production from food waste leachate, using a microbial pathway, is one of the most efficient technologies, due to its eco-friendly nature and high energy yield. Therefore, the present study aimed to evaluate the ability of an enriched bacterial mixture, isolated from forest soil, to enhance hydrogen production from food waste leachate using biochar. A lab-scale analysis was conducted at 35 °C and at different pH values (4, no adjustment, 6, 6.5, 7, and 7.5) over a period of 15 days. The sample with the enriched bacterial mixture supplemented with an optimum of 10 g/L of biochar showed the highest performance, with a maximum hydrogen yield of 1620 mL/day on day three. The total solid and volatile solid removal rates were 78.5% and 75% after 15 days, respectively. Acetic and butyrate acids were the dominant volatile fatty acids produced during the process, as favorable metabolic pathways for accelerating hydrogen production.

Investigation of Lipolytic-Secreting Bacteria from an Artificially Polluted Soil Using a Modified Culture Method and Optimization of Their Lipase Production.

Compared to lipases from plants or animals, microbial lipases play a vital role in different industrial applications and biotechnological perspectives due to their high stability and cost-effectiveness. Therefore, numerous lipase producers have been investigated in a variety of environments in the presence of lipidic carbon and organic nitrogen sources. As a step in the development of cultivating the unculturable functional bacteria in this study, the forest soil collected from the surrounding plant roots was used to create an artificially contaminated environment for lipase-producing bacterial isolation. The ten strongest active bacterial strains were tested in an enzyme assay supplemented with metal ions such as Ca2+, Zn2+, Cu2+, Fe2+, Mg2+, K+, Co2+, Mn2+, and Sn2+ to determine bacterial tolerance and the effect of these metal ions on enzyme activity. Lipolytic bacteria in this study tended to grow and achieved a high lipase activity at temperatures of 35-40 °C and at pH 6-7, reaching a peak of 480 U/mL and 420 U/mL produced by Lysinibacillus PL33 and Lysinibacillus PL35, respectively. These potential lipase-producing bacteria are excellent candidates for large-scale applications in the future.

Description of antibiotic-producing novel bacteria Paraburkholderia antibiotica sp. nov. and Paraburkholderia polaris sp. nov.

Two white colony-forming, Gram-stain-negative, non-sporulating and motile bacteria, designated G-4-1-8T and RP-4-7T, were isolated from forest soil and Arctic soil, respectively. Both strains showed antimicrobial activity against Gram-negative pathogens (Pseudomonas aeruginosa and Escherichia coli) and could grow at a pH range of pH 4.0-11.0 (optimum, pH 7.0-9.0). Phylogenetic analyses based on their 16S rRNA gene sequences indicated that strains G-4-1-8T and RP-4-7T formed a lineage within the family Burkholderiaceae and were clustered as members of the genus Paraburkholderia. Strain G-4-1-8T showed the highest 16S rRNA sequence similarity to Paraburkholderia monticola JC2948T (98.1 %), while strain RP-4-7T showed the highest similarity to Paraburkholderia metrosideri DNBP6-1T (98.8 %). The only respiratory quinone in both strains was ubiquinone Q-8. Their principal cellular fatty acids were C16 : 0, cyclo-C17 : 0, summed feature 3 (iso-C15 :0 2-OH and/or C16 :1 ω7c) and summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c). Their major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol and an unidentified aminophospholipid. The DNA G+C content of strains G-4-1-8T and RP-4-7T were 63.7 and 61.3 mol%, respectively, while their genome lengths were 7.44 and 9.67 Mb, respectively. The genomes of both strains showed at least 12 putative biosynthetic gene clusters. The average nucleotide identity and in silico DNA-DNA hybridization relatedness values between both strains and most closely related Paraburkholderia species were below the species threshold values. Based on a polyphasic study, these isolated strains represent novel species belonging to the genus Paraburkholderia, for which the names Paraburkholderia antibiotica sp. nov. (G-4-1-8T= KACC 21617T=NBRC 114603T) and Paraburkholderia polaris sp. nov. (RP-4-7T=KACC 21621T=NBRC 114605T) are proposed.

Insights into the biodegradation of diesel oil and changes in bacterial communities in diesel-contaminated soil as a consequence of various soil amendments.

Soil amendment is a promising strategy to enhance biodegradation capacity of indigenous bacteria. To assess the consequences of various soil amendments before large-scale implementation, a microcosm study was employed to investigate the effects of nutrients (TN), surfactants (TS), oxidants (TO), biochar (TB), and zero-valent iron nanoparticles (nZVI; TNP) on diesel degradation, bacterial communities, and community-level physiological profiles (CLPPs) of legacy field contaminated soil. The results showed that the TN, TB, TNP, TS, and TO, reduced 75.8%, 63.9%, 62.8%, 49.3%, and 40.1% of total petroleum hydrocarbons (TPH), respectively, within 120 days, while control (TW) reduced only 33.8%. In all soil amendments, TPH reduction was positively correlated with oxidation-reduction potential and heterotrophic and TPH-degrading bacteria, while negatively correlated with total nitrogen and available phosphate. Furthermore, in TW, TB, and TNP microcosms, TPH reduction showed positive association with pH, whereas in TN, TS, and TO, TPH reduction was negatively associated with pH. The bacterial diversity was reduced in all treatments as a function of the soil amendment and remediation time: the enriched potential TPH-degrading bacteria were Dyella, Paraburkholderia, Clavibacter, Arthrobacter, Rhodanobacter, Methylobacterium, and Pandoraea. The average well colour development (AWCD) values in CLPPs were higher in TB, sustained and improved in TN, and markedly lower in TNP, TS, and TO microcosms. Overall, these data demonstrate that nutrients and biochar amendments may be helpful in boosting biodegradation, increasing diesel-degrading bacteria, and improving soil physiological functions. In conclusion, diesel degradation efficiency and bacterial communities are widely affected by both type and duration of soil amendments.

Azohydromonas caseinilytica sp. nov., a Nitrogen-Fixing Bacterium Isolated From Forest Soil by Using Optimized Culture Method.

A bacterial strain, designated strain G-1-1-14T, was isolated from Kyonggi University forest soil during a study of previously uncultured bacterium. The cells of strain G-1-1-14T were motile by means of peritrichous flagella, Gram-stain-negative, rod-shaped, and able to grow autotrophically with hydrogen and fix nitrogen. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain G-1-1-14T belonged to the genus Azohydromonas. The closest species of strain G-1-1-14T were Azohydromonas ureilytica UCM-80 T (98.4% sequence similarity), Azohydromonas lata IAM 12599 T (97.5%), Azohydromonas riparia UCM-11 T (97.1%), and Azohydromonas australica IAM 12664 T (97.0%). The genome of strain G-1-1-14T was 6,654,139 bp long with 5,865 protein-coding genes. The genome consisted of N2-fixing genes (nifH) and various regulatory genes for CO2 fixation and H2 utilization. The principal respiratory quinone was ubiquinone-8, and the major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, and phosphatidylglycerol. The major fatty acids were summed feature 3 (iso-C15 : 0 2-OH and/or C16 : 1 ω7c), C16 : 0, summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c), and cyclo-C17 : 0. The DNA G + C content was 69.9%. The average nucleotide identity (OrthoANI), in silico DNA-DNA hybridization (dDDH), and conventional DDH relatedness values were below the species demarcation values for novel species. Based on genomic, genetic, phylogenetic, phenotypic, and chemotaxonomic characterizations, strain G-1-1-14T represents a novel species within the genus Azohydromonas, for which the name Azohydromonas caseinilytica sp. nov. is proposed. The type strain is G-1-1-14T (= KACC 21615 T = NBRC 114390 T ).

Cold-shock gene cspC in the genome of Massilia polaris sp. nov. revealed cold-adaptation.

A straw coloured, motile and Gram-stain-negative bacterium, designated RP-1-19T was isolated from soil of Arctic station, Svalbard, Norway. Based on the phylogenetic analysis of its 16S rRNA gene sequence, strain RP-1-19T formed a lineage within the family Oxalobacteraceae and clustered together within the genus Massilia. The closest members were M. violaceinigra B2T (98.6% sequence similarity), M. eurypsychrophilia JCM 30074T (98.3%) and M. atriviolacea SODT (98.1%). The only respiratory quinone was ubiquinone-8. The principal cellular fatty acids were summed feature 3 (iso-C15:0 2-OH/C16:1ω7c) and C16:0. The major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylglycerol. The DNA G + C content of the type strain was 63.2%. The average nucleotide identity and in silico DNA-DNA hybridization values between strain RP-1-19T and closest members were ≤ 80 and 23.2%, respectively. The genome was 4,522,469 bp long with 30 scaffolds and 4076 protein-coding genes. The genome showed eight putative biosynthetic gene clusters responsible for various secondary metabolites. Genome analysis revealed the presence of cold-shock proteins CspA and CspC. Presence of cspA and cspC genes in the genome manifest ecophysiology of strain RP-1-19T that may help in cold-adaptation. Based on these data, strain RP-1-19T represents a novel species in the genus Massilia, for which the name Massilia polaris sp. nov. is proposed. The type strain is RP-1-19T (= KACC 21619T = NBRC 114359T).

Genome insight and description of antibiotic producing Massilia antibiotica sp. nov., isolated from oil-contaminated soil.

An ivory-coloured, motile, Gram-stain-negative bacterium, designated TW-1T was isolated from oil-contaminated experimental soil in Kyonggi University. The phylogenetic analysis based on 16S rRNA gene sequence revealed, strain TW-1T formed a lineage within the family Oxalobacteraceae and clustered as members of the genus Massilia. The closest members were M. pinisoli T33T (98.8% sequence similarity), M. putida 6NM-7T (98.6%), M. arvi THG-RS2OT (98.5%), M. phosphatilytica 12-OD1T (98.3%) and M. niastensis 5516S-1T (98.2%). The sole respiratory quinone is ubiquinone-8. The major cellular fatty acids are hexadeconic acid, cis-9, methylenehexadeconic acid, summed feature 3 and summed feature 8. The major polar lipids are phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylglycerol. The DNA G + C content of the type strain is 66.3%. The average nucleotide identity (ANI) and in silico DNA-DNA hybridization (dDDH) relatedness values between strain TW-1T and closest members were below the threshold value for species demarcation. The genome size is 7,051,197 bp along with 46 contigs and 5,977 protein-coding genes. The genome showed 5 putative biosynthetic gene clusters (BGCs) that are responsible for different secondary metabolites. Cluster 2 showed thiopeptide BGC with no known cluster blast, indicating TW-1T might produce novel antimicrobial agent. The antimicrobial assessment also showed that strain TW-1T possessed inhibitory activity against Gram-negative pathogens (Escherichia coli and Pseudomonas aeruginosa). This is the first report of the species in the genus Massilia which produces antimicrobial compounds. Based on the polyphasic study, strain TW-1T represents novel species in the genus Massilia, for which the name Massilia antibiotica sp. nov. is proposed. The type strain is TW-1T (= KACC 21627T = NBRC 114363T).