Bioprospecting potentials of moderately halophilic bacteria and the isolation of squalene producers from Kuwait sabkha.

Sabkhas in Kuwait are unique hypersaline marine environments under-explored for bacterial community composition and bioprospecting. The 16S rRNA sequence analysis of 46 isolates with distinct morphology from two Kuwait sabkhas recovered 11 genera. Phylum Firmicutes dominated these isolates, and Bacillus (32.6%) was recovered as the dominant genera, followed by Halococcus (17.4%). These isolates were moderately halophilic, and some of them showed tolerance and growth at extreme levels of salt (20%), pH (5 and/or 11), and temperature (55 °C). A higher percentage of isolates harbored protease (63.0), followed by DNase (41.3), amylase (41.3), and lipase (32.6). Selected isolates showed antimicrobial activity against E. faecalis and isolated Halomonas shengliensis, and Idiomarina piscisalsi harbored gene coding for dNDP-glucose 4,6-dehydratase (Glu 1), indicating their potential to produce biomolecules with deoxysugar moieties. Palmitic acid or oleic acid was the dominant fatty acid, and seven isolates had some polyunsaturated fatty acids (linolenic or γ-linolenic acid). Interestingly, six isolates belonging to Planococcus and Oceanobacillus genus produced squalene, a bioactive isoprenoid molecule. Their content increased 30-50% in the presence of Terbinafine. The potential bioactivities and extreme growth conditions make this untapped bacterial diversity a promising candidate for future bioprospecting studies.

Indiicoccus explosivorum gen. nov., sp. nov., isolated from an explosives waste contaminated site.

A pink-pigmented, Gram-stain-positive, aerobic, coccoid-shaped bacterial strain, designated as S5-TSA-19T, was isolated from an explosives contaminated site in Panchkula, Haryana, India. The 16S rRNA gene sequencing blast analysis indicated that the strain is a member of the family Planococcaceae with the highest sequence similarity to Planomicrobium soli XN13T (96.1 %), followed by Planococcus maitriensis S1T (95.6 %), Planococcus plakortidis DSM 23997T (95.6 %), Planomicrobium flavidum ISL-41T (95.6 %), Planococcus rifietoensis M8T (95.5 %), Planococcus salinus LCB217T (95.5 %) and Planococcus maritimus DSM 17275T (95.5 %). Phylogenetic analysis based on 16S rRNA gene and whole-genome sequences (based on a conserved set of 400 proteins) retrieved the strain in a distinct branch indicating a separate lineage within the family Planococcaceae. Strain S5-TSA-19T had a distinctive chemotaxonomic pattern comprising A4α type peptidoglycan based on l-Lys-d-Asp, iso-C15 : 0 as the major fatty acid, absence of phosphatidylethanolamine as a major lipid and MK-7 and MK-6 as the major menaquinones, differentiating it from the genera Planococcus and Planomicrobium, thus supporting the findings of molecular phylogeny. Further, strain S5-TSA-19T was able to biotransform hexahydro-1,3,5,-trinitro-1,2,5-triazine (RDX) into nitrite derivatives under aerobic conditions in 2-4 days, whereas the closest reference strains did not possess this property. On the basis of polyphasic taxonomic characterization and a phylogenomics approach, strain S5-TSA-19T is proposed as the type strain of a novel species in a novel genus for which the name Indiicoccus explosivorum gen. nov., sp. nov. is proposed (=JCM 31737T=KCTC 33871T=MTCC 12608T).

Planomicrobium iranicum sp. nov., a novel slightly halophilic bacterium isolated from a hypersaline wetland.

A novel strain, designated as MX6T was isolated from Meighan wetland, in the centre of Iran. The cells were Gram-stain-positive, motile, coccoid to rod-shaped, oxidase- and catalase-positive. The strain grew optimally at 35 °C, 3 % (w/v) NaCl and pH 7-7.5. A polyphasic taxonomic study was undertaken in order to characterize the strain in detail. The results of phylogenetic analysis based on 16S rRNA gene sequences indicated that MX6T represented a member of the phylum Firmicutes, family Planococcaceae, genus Planomicrobium, and showed the highest similarity with Planomicrobium flavidum ISL-41T (98.2 %) and Planomicrobium psychrophilum CMC 53orT (98.0 %). The main polar lipids of MX6T consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and seven unidentified phospholipids and its DNA G+C content was 45.5 mol%. Major cellular fatty acids were anteiso-C15 : 0, C16 : 1ω7c alcohol, iso-C14 : 0, iso-C15 : 0 and iso-C16 : 0 and the predominant respiratory quinone was Q-8 (62 %). Experimental DNA-DNA hybridization between MX6T and Planomicrobium flavidum IBRC-M 11047T was 20 %, supporting the differential taxonomic status of MX6T as representing a different taxon. All these data indicate that MX6T represents a novel species of the genus Planomicrobium, for which the name Planomicrobium iranicum sp. nov. is proposed. The type strain is MX6T (=IBRC M 10928T=LMG 28548T).

Jeotgalibacillus proteolyticus sp. nov., a protease-producing bacterium isolated from ocean sediments.

A Gram-stain-positive, rod-shaped bacterial strain, 22-7T, was isolated from ocean sediment of Laizhou Bay, China, and was characterized by using a polyphasic approach. Optimal growth was observed at 33 °C on a 2216E agar plate of pH 7.5 and with 2 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences identified it as a member of the genus Jeotgalibacillus, most similar to Jeotgalibacillus campisalis SF-57T (98.7 % similarity), Jeotgalibacillus marinus DSM 1297T (98.2 %) and Jeotgalibacillus soli P9T (97.1 %). Average nucleotide identity values and digital DNA-DNA hybridization values were less than 74.2 and 18.1 %, respectively, between strain 22-7T and the type strains of closely related species. The major polar lipids were aminophospholipid, phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol; the major fatty acids (>10 %) were anteiso-C15 : 0 and iso-C15 : 0; and the major menaquinone was MK-7. The peptidoglycan type of the cell wall was A1α linked through l-lysine as the diamino acid. Combined data from phenotypic, chemotaxonomic and genotypic characterizations demonstrated that strain 22-7T represents a novel Jeotgalibacillus species, for which the name Jeotgalibacillus proteolyticus sp. nov. is proposed. The type strain is 22-7T(=MCCC 1H00228T=KCTC 33930T).

Bhargavaea changchunensis sp. nov. isolated from soil in China.

A Gram-staining-positive, aerobic, rod-shaped (201802YP6T) bacteria was isolated from soil, Northeast of China. Growth occurred at 10-40 °C (optimum 25-30 °C), at pH 6.0-8.0 (optimum 7.0) and at 0-2% NaCl. Based on 16S rRNA gene sequence analysis, the nearest phylogenetic neighbors of strain 201802YP6T were identified as Bhargavaea cecembensis DSE10T (99.52%), Bhargavaea beijingensis ge10T (99.45%), Bhargavaea indica KJW98T (99.45%), Bhargavaea ullalensis ZMA19T (98.81%), and Bhargavaea ginsengi ge14T (98.76%). Levels of similarity among strain 201802YP6T and other Bhargavaea species were lower than 98.76%. GyrB amino acid sequence-based analysis supported the phylogenetic position and also distinguished strain 201802YP6T from the other species of the genus Bhargavaea. DNA-DNA hybridization values between strain 201802YP6T and B. cecembensis, B. beijingensis, B. indica, B. ullalensis, B. ginsengi were 43.5%, 43%, 32.5%, 30.5% and 20.4%, respectively. The DNA G + C content of strain 201802YP6T was 51.23 mol%. The average nucleotide identity (ANI) of the draft genome was 87.04% to B. cecembensis DSE10T. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, glycolipids, phosphatidylethanolamine, and phosphatidyllipid. The predominant menaquinone was MK-8. The major fatty acids were iso-C15:0 (39.91%), anteiso-C15:0 (28.86%), anteiso-C17:0 (6.30%) and C16:0 (6.13%). On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics and DNA-DNA hybridization data, strain 201802YP6T represents a novel species of the genus Bhargavaea, for which the name Bhargavaea changchunensis sp. nov. is proposed. The type strain is 201802YP6T (= CGMCC 1.13508T = KCTC 33975T).

Kurthia ruminicola sp. nov., isolated from the rumen contents of a Holstein cow.

Gram-staining-positive, motile, rod-shaped bacteria, designated as H31022T and H31024 was isolated from rumen contents of a Holstein cow. Optimum growth occurred at 25°C and pH 7.0 on R2A agar medium. Oxidase and catalase activities are positive. The 16S rRNA gene sequence (1,452 bp) of the new isolates revealed they belong to the genus Kurthia of the phylum Firmicutes. Highest gene sequence similarities were assessed to be with Kurthia massiliensis JC30T (98.4%), Kurthia senegalensis JC8ET (97.5%), and Kurthia populi 10y-14T (97.4%). Kurthia sibirica DSM 4747T (97.3%), Kurthia zopfii NBRC 101529T (97.0%), and Kurthia gibsonii NCIMB 9758T (96.7%). DNA G + C content of strains H31022T and H31024 were 34.4% and 39.7%. Strains H31022T and H31024 has the following chemotaxonomic characteristics; the major fatty acids are iso-C15:0, iso-C14:0 and anteiso-C15; polar lipid profile contained diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), unknown aminophospholipids (APL), unknown glycolipids (GL), unknown phospholipids (PL), and unknown polar lipids (L); the major quinone is MK-7. Based on polyphasic taxonomic analysis, strains H31022T (= KCTC 33923T = JCM 19640T) and H31024 (= KCTC 33924T = JCM 19641T) identified a novel species in the genus Kurthia for which the name Kurthia ruminicola sp. nov. is proposed.

Solibacillus kalamii sp. nov., isolated from a high-efficiency particulate arrestance filter system used in the International Space Station.

A Gram-stain-positive, rod-shaped, endospore-forming, aerobic bacterial strain, designated ISSFR-015T, was isolated from a high-efficiency particulate arrestance filter in the International Space Station and was characterized by polyphasic taxonomy. A comparative analysis of the 16S rRNA gene sequence (1494 bp) of strain ISSFR-015T showed highest similarity to Solibacillus isronensis B3W22T (98.9 %), followed by Solibacillus silvestris HR3-23T (98.6 %) and Bacillus cecembensis PN5T (96.7 %). DNA-DNA hybridization analysis revealed that the DNA relatedness values of strain ISSFR-015T with other closely related species were in the range of 41-47 % [S. silvestrisMTCC 10789T (47 %), S. isronensis MTCC 7902T (41 %) and B. cecembensis MTCC 9127T (43 %)]. The DNA G+C content of strain ISSFR-015T was 45.4 mol%. The major fatty acids were iso-C15 : 0 (45.2 %) and C17 : 1ω10c (12.1 %). The polar lipid profile contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylserine and one unknown phospholipid. The isoprenoid quinones present in strain ISSFR-015T were MK-7 (86.8 %), MK-6 (11.6 %) and MK-8 (1.0 %). The peptidoglycan type of the cell wall was A4α l-Lys-d-Glu. Based on the phylogenetic analysis, strain ISSFR-015T belongs to the genus Solibacillus. The polyphasic taxonomic data, including low DNA-DNA hybridization values, and the chemotaxonomic analysis confirmed that strain ISSFR-015T represents a novel species, for which the name Solibacillus kalamii sp. nov. is proposed. The type strain for this proposed species is ISSFR-015T (=NRRL B-65388T=DSM 101595T).

Savagea faecisuis gen. nov., sp. nov., a tylosin- and tetracycline-resistant bacterium isolated from a swine-manure storage pit.

A polyphasic taxonomic study using morphological, biochemical, chemotaxonomic and molecular methods was performed on three strains of a Gram-stain positive, non-sporeforming, motile aerobic rod-shaped bacterium resistant to tylosin and tetracycline isolated from a swine-manure storage pit. On the basis of 16S rRNA gene sequence analyses, it was confirmed that these isolates are highly related to each other and form a hitherto unknown lineage within the Planococcaceae. In particular, pairwise analysis of the 16S rRNA gene sequence demonstrated that the novel organism is closely related to members of the genus Sporosarcina (92.8-94.5 %), Pyschrobacillus (93.5-93.9 %) and Paenisporosarcina (93.3-94.5 %). The predominant fatty acids were found to consist of iso-C15:0 and iso-C17:1 ω10c and the G+C mol% was determined to be 41.8. Based on biochemical, chemotaxonomic, and phylogenetic evidence, it is proposed that these novel strains be classified as a novel genus and species, Savagea faecisuis gen nov., sp. nov. The type strain is Con12(T) (=CCUG 63563(T) = NRRL B-59945(T) = NBRC 109956(T)).

Isolation of Jeotgalibacillus malaysiensis sp. nov. from a sandy beach, and emended description of the genus Jeotgalibacillus.

A Gram-stain-positive, endospore-forming, rod-shaped bacterial strain, designated D5(T), was isolated from seawater collected from a sandy beach in a southern state of Malaysia and subjected to a polyphasic taxonomic study. Sequence analysis of the 16S rRNA gene demonstrated that this isolate belongs to the genus Jeotgalibacillus, with 99.87% similarity to Jeotgalibacillus alimentarius JCM 10872(T). DNA-DNA hybridization of strain D5(T) with J. alimentarius JCM 10872(T) demonstrated 26.3% relatedness. The peptidoglycan type was A1α linked directly to L-lysine as the diamino acid. The predominant quinones identified in strain D5(T) were menaquinones MK-7 and MK-8.The major fatty acids were iso-C15:0 and anteiso-C15:0. The G+C content of its DNA was 43.0 mol%. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and sulfoquinovosyl diacylglycerol, as well as two unknown phospholipids and three unknown lipids. The phenotypic, chemotaxonomic and genotypic data indicated that strain D5(T) represents a novel species of the genus Jeotgalibacillus, for which the name Jeotgalibacillus malaysiensis sp. nov. is proposed (type strain D5(T) = DSM 28777(T) = KCTC33550(T)). An emended description of the genus Jeotgalibacillus is also provided.

Chryseomicrobium aureum sp. nov., a bacterium isolated from activated sludge.

A Gram-stain-positive, rod-shaped, non-motile, non-spore-forming, aerobic bacterial strain, designated BUT-2(T), was isolated from activated sludge of one herbicide-manufacturing wastewater-treatment facility in Kunshan, Jiangsu province, China, and subjected to polyphasic taxonomic studies. Analysis of the 16S rRNA gene sequence indicated that strain BUT-2(T) shared the highest similarity with Chryseomicrobium amylolyticum (98.98%), followed by Chryseomicrobium imtechense (98.88%), with less than 96% similarlity to members of the genera Paenisporosarcina, Planococcus, Sporosarcina and Planomicrobium. Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain BUT-2(T) clustered with C. amylolyticum JC16(T) and C. imtechense MW10(T), occupying a distinct phylogenetic position. The major fatty acid (>10% of total fatty acids) type of strain BUT-2(T) was iso-C(15 : 0). The quinone system comprised menaquinone MK-7 (77.8%), MK-6 (11.9%) and MK-8 (10.3%). The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and some unidentified phospholipids. The cell-wall peptidoglycan type of strain BUT-2(T) was L-Orn-D-Glu. The genomic DNA G+C content of strain BUT-2(T) was 48.5 mol%. Furthermore, the DNA-DNA relatedness in hybridization experiments against the reference strain was lower than 70%, confirming that strain BUT-2(T) did not belong to previously described species of the genus Chryseomicrobium. On the basis of its morphological, physiological and chemotaxonomic characteristics as well as phylogenetic analysis, strain BUT-2(T) is considered to represent a novel species of the genus Chryseomicrobium, for which the name Chryseomicrobium aureum sp. nov. is proposed. The type strain is BUT-2(T) ( = CCTCC AB2013082(T) = KACC 17219(T)).

Planomicrobium soli sp. nov., isolated from soil.

A Gram-staining-positive bacterium, designated strain XN13(T), was isolated from a soil sample collected from ALaShan National Geological Park in Inner Mongolia Autonomous Region, China and subjected to a taxonomic study using a polyphasic approach. Strain XN13(T) was found to have a range of chemical and morphological properties consistent with its classification in the genus Planomicrobium. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain XN13(T) was related to members of the genus Planomicrobium. The closest phylogenetic relatives were Planomicrobium okeanokoites NBRC 12536(T), Planomicrobium koreense JG07(T), Planomicrobium mcmeekinii S23F2(T) and Planomicrobium flavidum ISL-41(T) with 98.2%, 97.8%, 97.8% and 97.7% 16S rRNA gene sequence similarity, respectively. The major cellular fatty acids were anteiso-C(15 : 0), C(16 : 1)ω7c alcohol, iso-C(14 : 0) and C(16 : 1)ω11c. The predominant menaquinones were MK-8 and MK-7. The DNA G+C content was 40.3 mol%. The DNA-DNA relatedness values between strain XN13(T) and Planomicrobium okeanokoites KCTC 3672(T), Planomicrobium koreense KCTC 3684(T), P. mcmeekinii CGMCC 1.2724(T), Planomicrobium flavidum KCTC 13261(T), Planomicrobium chinense CGMCC 1.3454(T) and Planomicrobium glaciei CGMCC 1.6846(T) were 36%, 30%, 34%, 29%, 30% and 31%, respectively. The organism is different from recognized species of the genus Planomicrobium in several phenotypic characteristics. On the basis of phenotypic and genotypic properties, strain XN13(T) represents a novel species of the genus Planomicrobium, for which the name Planomicrobium soli sp. nov. is proposed. The type strain is XN13(T) ( = CGMCC 1.12259(T) = KCTC 33047(T)).

Chryseomicrobium imtechense gen. nov., sp. nov., a new member of the family Planococcaceae.

A Gram-stain-positive, rod-shaped, yellow, non-motile, non-spore-forming, strictly aerobic bacterial strain, designated MW 10(T), was isolated from seawater of the Bay of Bengal, India, and was subjected to a polyphasic taxonomic study. Analysis of the 16S rRNA gene sequence revealed that strain MW 10(T) showed highest similarity to the type strains of Psychrobacillus psychrodurans (96.15 %) and Psychrobacillus psychrotolerans (96.01 %) and showed less than 96 % similarity to members of the genera Paenisporosarcina, Planococcus, Sporosarcina and Planomicrobium. Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain MW 10(T) formed a clade separate from members of closely related genera. The morphological, physiological and chemotaxonomic characteristics of strain MW 10(T) differed from those of members of closely related genera. The major fatty acid in strain MW 10(T) was iso-C(15 : 0) and the menaquinones were MK-7 (48.4 %), MK-8 (32.3 %), MK-7(H(2)) (13.7 %) and MK-6 (5.6 %). The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, an unknown phospholipid, an unknown lipid and an unknown glycolipid. The cell-wall peptidoglycan type was l-Lys-d-Asp. The genomic DNA G+C content (53.4 mol%) of strain MW 10(T) was significantly different from those of members of closely related genera. On the basis of its morphological, physiological and chemotaxonomic characteristics as well as our phylogenetic analysis, we conclude that strain MW 10(T) is a member of a novel genus and species, for which the name Chryseomicrobium imtechense gen. nov., sp. nov., is proposed. The type strain of Chryseomicrobium imtechense is MW 10(T) ( = MTCC 10098(T)  = JCM 16573(T)).