Spatial and Temporal Dynamics of Prokaryotic and Viral Community Assemblages in a Lotic System (Manatee Springs, Florida).

Flow from high-magnitude springs fed by the Floridan aquifer system contributes hundreds of liters of water per second to rivers, creating unique lotic systems. Despite their importance as freshwater sources and their contributions to the state's major rivers, little is known about the composition and spatiotemporal variability of prokaryotic and viral communities of these spring systems or their influence on downstream river sites. At four time points throughout a year, we determined the abundance and diversity of prokaryotic and viral communities at three sites within the first-magnitude Manatee Springs system (the spring head where water emerges from the aquifer, a mixed region where the spring run ends, and a downstream site in the Suwannee River). The abundance of prokaryotes and virus-like particles increased 100-fold from the spring head to the river and few members from the head communities persisted in the river at low abundance, suggesting the springs play a minor role in seeding downstream communities. Prokaryotic and viral communities within Manatee Springs clustered by site, with seasonal variability likely driven by flow. As water flowed through the system, microbial community composition was affected by changes in physiochemical parameters and community coalescence. Evidence of species sorting and mass effects could be seen in the assemblages. Greater temporal fluctuations were observed in prokaryotic and viral community composition with increasing distance from the spring outflow, reflecting the relative stability of the groundwater environment, and comparisons to springs from prior work reaffirmed that distinct first-magnitude springs support unique communities. IMPORTANCE Prokaryotic and viral communities are central to food webs and biogeochemical processes in aquatic environments, where they help maintain ecosystem health. The Floridan aquifer system (FAS), which is the primary drinking water source for millions of people in the southeastern United States, contributes large amounts of freshwater to major river systems in Florida through its springs. However, there is a paucity of information regarding the spatiotemporal dynamics of microbial communities in these essential flowing freshwater systems. This work explored the prokaryotic and viral communities in a first-magnitude spring system fed by the FAS that discharges millions of liters of water per day into the Suwannee River. This study examined microbial community composition through space and time as well as the environmental parameters and metacommunity assembly mechanisms that shape these communities, providing a foundational understanding for monitoring future changes.

Genomes of Novel Myxococcota Reveal Severely Curtailed Machineries for Predation and Cellular Differentiation.

Cultured Myxococcota are predominantly aerobic soil inhabitants, characterized by their highly coordinated predation and cellular differentiation capacities. Little is currently known regarding yet-uncultured Myxococcota from anaerobic, nonsoil habitats. We analyzed genomes representing one novel order (o__JAFGXQ01) and one novel family (f__JAFGIB01) in the Myxococcota from an anoxic freshwater spring (Zodletone Spring) in Oklahoma, USA. Compared to their soil counterparts, anaerobic Myxococcota possess smaller genomes and a smaller number of genes encoding biosynthetic gene clusters (BGCs), peptidases, one- and two-component signal transduction systems, and transcriptional regulators. Detailed analysis of 13 distinct pathways/processes crucial to predation and cellular differentiation revealed severely curtailed machineries, with the notable absence of homologs for key transcription factors (e.g., FruA and MrpC), outer membrane exchange receptor (TraA), and the majority of sporulation-specific and A-motility-specific genes. Further, machine learning approaches based on a set of 634 genes informative of social lifestyle predicted a nonsocial behavior for Zodletone Myxococcota. Metabolically, Zodletone Myxococcota genomes lacked aerobic respiratory capacities but carried genes suggestive of fermentation, dissimilatory nitrite reduction, and dissimilatory sulfate-reduction (in f_JAFGIB01) for energy acquisition. We propose that predation and cellular differentiation represent a niche adaptation strategy that evolved circa 500 million years ago (Mya) in response to the rise of soil as a distinct habitat on Earth. IMPORTANCE The phylum Myxococcota is a phylogenetically coherent bacterial lineage that exhibits unique social traits. Cultured Myxococcota are predominantly aerobic soil-dwelling microorganisms that are capable of predation and fruiting body formation. However, multiple yet-uncultured lineages within the Myxococcota have been encountered in a wide range of nonsoil, predominantly anaerobic habitats, and the metabolic capabilities, physiological preferences, and capacity of social behavior of such lineages remain unclear. Here, we analyzed genomes recovered from a metagenomic analysis of an anoxic freshwater spring in Oklahoma, USA, that represent novel, yet-uncultured, orders and families in the Myxococcota. The genomes appear to lack the characteristic hallmarks for social behavior encountered in Myxococcota genomes and displayed a significantly smaller genome size and a smaller number of genes encoding biosynthetic gene clusters, peptidases, signal transduction systems, and transcriptional regulators. Such perceived lack of social capacity was confirmed through detailed comparative genomic analysis of 13 pathways associated with Myxococcota social behavior, as well as the implementation of machine learning approaches to predict social behavior based on genome composition. Metabolically, these novel Myxococcota are predicted to be strict anaerobes, utilizing fermentation, nitrate reduction, and dissimilarity sulfate reduction for energy acquisition. Our results highlight the broad patterns of metabolic diversity within the yet-uncultured Myxococcota and suggest that the evolution of predation and fruiting body formation in the Myxococcota has occurred in response to soil formation as a distinct habitat on Earth.

Trichococcus shcherbakoviae subsp. psychrophilus subsp. nov., a psychrotolerant facultative anaerobe isolated from a cold spring.

A psychrotolerant facultative anaerobe, strain SKBGT, was isolated from the bottom sediments of the cold mineral spring Buxichen (Buryatia, Russia). Gram-positive non-motile cocci with a diameter of 1.75-2.5 µm were observed singly or in long chains. Cells grew in the temperature range from ̶ 5-35 °C. Growth was observed within the pH range of 7.0-9.5, with the optimum growth at pH 7.6 and at a NaCl concentration from 0-1.0 % (optimum 0.1 % (w/v)). Strain SKBGT was a chemoorganoheterotroph that used sugars and some organic acids as substrates. The predominant fatty acids in cell walls were С16:1ω9, С18:1ω9, and С16 : 0. The 16S rRNA gene sequence of strain SKBGT shared high similarity (>99 %) with those of the type strains of the genus Trichococcus. Digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values between strain SKBGT and Trichococcus shcherbakoviae ArtT (=DSM 107162T=VKM B-3260T) were 70.1 and 95.4 %, respectively. The genomic DNA G+C content of strain SKBGT was 47.1 mol%. Compared with the type strain of T. shcherbakoviae, the new strain was characterized by a temperature optimum for growth (10 °C) significantly lower than that of T. shcherbakoviae DSM 107162T (20-30 °C). Based on phenotypic and genomic characteristics, the isolate SKBGT was classified as T. shcherbakoviae subsp. psychrophilus subsp. nov. The type strain is SKBGT (=VKM B-3241Т=JCM 33326T).

Sphingobium algorifonticola sp. nov., isolated from a cold spring.

Strain TLA-22T, isolated from a cold spring in Taiwan, was characterized using a polyphasic taxonomy approach. Cells were Gram-stain-negative, aerobic, poly-ß-hydroxybutyrate-accumulating, motile by means of a single polar flagellum, rod-shaped and formed bright yellow colonies. Optimal growth occurred at 20-25 °C, pH 6-6.5, and in the presence of 0.5 % NaCl. The major fatty acids of TLA-22T were C18 : 1 ω7 c and C17 : 1ω6c. The predominant hydroxy fatty acids were C15 : 0 2-OH and C14 : 0 2-OH. The polar lipid profile consisted of a mixture of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidyldimethylethanolamine, sphingoglycolipid, an unidentified aminophospholipid, an unidentified phospholipid and three unidentified lipids. TLA-22T contained spermidine as the major polyamine and putrescine as the minor component. The only isoprenoid quinone was Q-10. The genomic DNA G+C content of TLA-22T was 63.2 mol%. Phylogenetic analyses based on 16S rRNA gene sequences and coding sequences of 92 protein clusters indicated that TLA-22T was a mem,ber of a phylogenetic lineage including members of the genus Sphingobium. TLA-22T was most closely related to Sphingobium aromaticiconvertens RW16T, with a 97.4 % 16S rRNA gene sequence similarity. TLA-22T showed 74.8-75.7 % average nucleotide identity and 20.1-22.0 % digital DNA-DNA hybridization identity with the strains of other species of the genus Sphingobium. On the basis of phenotypic and genotypic properties and phylogenetic inference, strain TLA-22T should be classified as representing a novel species of the genus Sphingobium, for which the name Sphingobium algorifonticola sp. nov. is proposed. The type strain is TLA-22T (=BCRC 81097T =LMG 30309T=KCTC 62189T).

The quality of drinking and domestic water from the surface water sources (lakes, rivers, irrigation canals and ponds) and springs in cholera prone communities of Uganda: an analysis of vital physicochemical parameters.

BACKGROUND: Water is the most abundant resource on earth, however water scarcity affects more than 40% of people worldwide. Access to safe drinking water is a basic human right and is a United Nations Sustainable Development Goal (SDG) 6. Globally, waterborne diseases such as cholera are responsible for over two million deaths annually. Cholera is a major cause of ill-health in Africa and Uganda. This study aimed to determine the physicochemical characteristics of the surface and spring water in cholera endemic communities of Uganda in order to promote access to safe drinking water. METHODS: A longitudinal study was carried out between February 2015 and January 2016 in cholera prone communities of Uganda. Surface and spring water used for domestic purposes including drinking from 27 sites (lakes, rivers, irrigation canal, springs and ponds) were tested monthly to determine the vital physicochemical parameters, namely pH, temperature, dissolved oxygen, conductivity and turbidity. RESULTS: Overall, 318 water samples were tested. Twenty-six percent (36/135) of the tested samples had mean test results that were outside the World Health Organization (WHO) recommended drinking water range. All sites (100%, 27/27) had mean water turbidity values greater than the WHO drinking water recommended standards and the temperature of above 17 °C. In addition, 27% (3/11) of the lake sites and 2/5 of the ponds had pH and dissolved oxygen respectively outside the WHO recommended range of 6.5-8.5 for pH and less than 5 mg/L for dissolved oxygen. These physicochemical conditions were ideal for survival of Vibrio. cholerae. CONCLUSIONS: This study showed that surface water and springs in the study area were unsafe for drinking and had favourable physicochemical parameters for propagation of waterborne diseases including cholera. Therefore, for Uganda to attain the SDG 6 targets and to eliminate cholera by 2030, more efforts are needed to promote access to safe drinking water. Also, since this study only established the vital water physicochemical parameters, further studies are recommended to determine the other water physicochemical parameters such as the nitrates and copper. Studies are also needed to establish the causal-effect relationship between V. cholerae and the physicochemical parameters.

Altererythrobacter aquimixticola sp. nov., isolated from sediment sampled at the junction between the ocean and a freshwater spring.

A Gram-stain-negative, aerobic, non-motile and coccoid-, ovoid- or rod-shaped bacterial strain, designated SSKS-13T, was isolated from sediment sampled at the junction between the ocean and a freshwater spring at Jeju island, Republic of Korea. Strain SSKS-13T grew optimally at 37 °C and in the presence of 2.0 % (w/v) NaCl. A neighbour-joining phylogenetic tree of 16S rRNA gene sequences showed that strain SSKS-13T fell within the clade comprising the type strains of Altererythrobacter species, clustering with the type strains of Altererythrobacter lauratis, Altererythrobacter palmitatis and Altererythrobacter buctensis having 16S rRNA gene sequence similarities of 97.2-97.6 %. Strain SSKS-13T exhibited 16S rRNA gene sequence similarities of less than 97.0 % to the type strains of the other recognized species. Strain SSKS-13T contained Q-10 as the predominant ubiquinone and C18 : 1 ω7c as the major fatty acid. The major polar lipids detected in strain SSKS-13T were phosphatidylethanolamine, phosphatidylglycerol and sphingoglycolipid. The DNA G+C content of strain SSKS-13T was 64.6 mol%. The mean DNA-DNA relatedness values of strain SSKS-13T with the type strains of A. lauratis, A. palmitatis and A. buctensis were 11.7-25.3 %. Differential phenotypic properties, together with the phylogenetic and genetic data, proved that strain SSKS-13T is distinct from recognized Altererythrobacter species. On the basis of the data presented here, strain SSKS-13T is considered to represent a novel species of the genus Altererythrobacter, for which the name Altererythrobacter aquimixticola sp. nov. is proposed. The type strain is SSKS-13T (=KACC 19863T=KCTC 62900T=NBRC 113545T).

Paracoccus fontiphilus sp. nov., isolated from a freshwater spring.

Strain MVW-1T, isolated from a freshwater spring in Taiwan, was characterized by using a polyphasic taxonomy approach. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain MVW-1T belongs to the genus Paracoccus and has the highest levels of sequence similarity to Paracoccus caeni MJ17T (97.6 %), Paracoccus sediminis CMB17T (97.4 %), Paracoccus angustae E6T (97.3 %) and Paracoccus acridae SCU-M53T (97.1 %). Cells were Gram-stain-negative, aerobic, poly-ß-hydroxybutyrate-accumulating, non-motile, rod-shaped and formed light orange-coloured colonies. Optimal growth occurred at 20-25 °C, pH 6-7, and in the presence of 0-3 % NaCl. The major fatty acid of strain MVW-1T was C18 : 1ω7c. The polar lipid profile consisted of phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, diphosphatidylglycerol, an unidentified glycolipid, an unidentified aminolipid and three unidentified phospholipids. The predominant polyamines were spermidine, putrescine and cadaverine. The only isoprenoid quinone was Q-10. The genomic DNA G+C content of strain MVW-1T was 63.4 mol%. Strain MVW-1T exhibited less than 35 % DNA-DNA relatedness to P. caeni MJ17T, P. angustae E6T, P. sediminis CMB17T and P. acridae SCU-M53T. On the basis of phenotypic and genotypic properties and phylogenetic inference, strain MVW-1T should be classified in a novel species of the genus Paracoccus, for which the name Paracoccus fontiphilus sp. nov. is proposed. The type strain is MVW-1T (=BCRC 80974T=LMG 29554T=KCTC 52239T).

Occultifur mephitis f.a., sp. nov. and other yeast species from hypoxic and elevated CO2 mofette environments.

An inventory of culturable yeasts from the soil and water of natural CO2 springs (mofettes) in northeast Slovenia is presented. In mofettes, CO2 of geological origin reaches the soil surface causing temporarily and spatially stable hypoxic environments in soil and water. In total, 142 yeast strains were isolated and identified from high CO2 and control meadow soil, meadow ground-water, forest pond and stream water. All water locations showed below-ground CO2 release. They were assigned to six basidiomycetous yeast genera (six species) and 11 ascomycetous genera (18 species). All ascomycetous yeasts, with the exception of Debaryomyces hansenii, were able to grow under elevated CO2 and fermented glucose. Candida sophiae-reginae, Pichia fermentans and Candida vartiovaarae were the dominating species in meadow and forest high CO2 exposed water. Meyerozyma guilliermondii and Wickerhamomyces anomalus predominated in high CO2 exposed soils. Using high dilution plating of a mofette soil sample, four strains of an unknown basidiomycetous species were isolated and are here newly described as Occultifur mephitis based on molecular phylogenetic and phenotypic criteria. The type strain of Occultifur mephitis is EXF-6436T[CBS 14611=PYCC 7049, LT594852 (D1/D2), KX929055 (ITS)]. An additional three isolated strains are EXF-6437 (LT594853, KX929056), EXF-6473 (LT594863, KX929057) and EXF-6482 (LT594867, KX929054), as well as a strain reported from previous studies isolated from a leaf of Cistus albidus in Portugal (CBS 10223=PYCC 6067), EU002842 (D1/D2), KY308183 (ITS).

Do natural spring waters in Australia and New Zealand affect health? A systematic review.

Therapeutic use of spring waters has a recorded history dating back to at least 1550 BC and includes both bathing in and drinking such waters for their healing properties. In Australia and New Zealand the use of therapeutic spring waters is a much more recent phenomenon, becoming a source of health tourism from the late 1800s. We conducted a systematic review aimed at determining the potential health outcomes relating to exposure to Australian or New Zealand natural spring water. We found only low-level evidence of adverse health outcomes relating to this spring water exposure, including fatalities from hydrogen sulphide poisoning, drowning and primary amoebic meningoencephalitis. We found no studies that investigated the therapeutic use of these waters, compared with similar treatment with other types of water. From the broader literature, recommendations have been made, including fencing potentially harmful spring water, and having signage and media messages to highlight the potential harms from spring water exposure and how to mitigate the risks (e.g. not putting your head under water from geothermal springs). Sound research into the potential health benefits of Australian and New Zealand spring waters could provide an evidence base for the growing wellness tourism industry.

Cellvibrio fontiphilus sp. nov., isolated from a spring.

A bacterial strain, designated MVW-40T, was isolated from Maolin Spring in Taiwan and characterized using a polyphasic taxonomy approach. Cells of strain MVW-40T were Gram-negative, strictly aerobic, motile by a single polar flagellum and bright yellow-pigmented rods with pointed ends. Growth occurred at 15-40 °C (optimum, 20-30 °C), at pH 6-9 (optimum, pH 6) and with 0-2 % NaCl (optimum, 0 %). Phylogenetic analyses based on 16S rRNA gene sequences showed that strain MVW-40T belonged to the genus Cellvibrio and showed the highest levels of sequence similarity with respect to Cellvibrio mixtussubsp. mixtus ACM 2601T (98.1 %) and Cellvibrio fibrivorans R-4079T (97.2 %). Strain MVW-40T contained summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), C16 : 0 and C18 : 1ω7c as the predominant fatty acids. The polar lipid profile consisted of phosphatidylethanolamine, phosphatidylglycerol, two uncharacterized aminophospholipids, two uncharacterized phospholipids and an uncharacterized lipid. The DNA G+C content of the genomic DNA was 52.8 mol%. The DNA-DNA hybridization value for strain MVW-40T with C. mixtussubsp. mixtus ACM 2601T and C. fibrivorans R-4079T was less than 45 %. On the basis of the phylogenetic inference and phenotypic data, strain MVW-40T should be classified as a novel species, for which the name Cellvibrio fontiphilus sp. nov. is proposed. The type strain is MVW-40T (=BCRC 80977T=LMG 29557T=KCTC 52237T).

Draft genome and description of Consotaella salsifontis gen. nov. sp. nov., a halophilic, free-living, nitrogen-fixing alphaproteobacterium isolated from an ancient terrestrial saline spring.

A free-living, nitrogen-fixing, mesophilic and facultative aerobe, designated strain USBA 369T, was isolated from a terrestrial saline spring of the Colombian Andes. The non-sporulating rods (1.5×0.8 µm) with rounded ends stained Gram-negative and were motile by means of lophotrichous flagella. The strain grew optimally at 30 °C, at pH 6.9-7.5 and with 1.5 % (w/v) NaCl. The major fatty acids detected were C18 : 1ω7c and C19 : 0 cyclo ω8c, and the respiratory lipoquinone ubiquinone 10 (Q-10) was present. The genome consisted of 4.65 Mb with a DNA G+C content of 64.3 mol%. A total of 4371 genes were predicted and, of those, 4300 were protein coding genes and 71 were RNA genes. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain USBA 369T formed a different lineage within the class Alphaproteobacteria, order Rhizobiales, and DNA homology studies with the most closely related genera, Aurantimonas, Aureimonas and Rhizobium (95 % 16S rRNA gene sequence similarity), showed values of <15 %. The phylogenomic analysis provided evidence for clear phylogenetic divergence between strain USBA 369T and the closely related genera. On the basis of the phenotypic, chemotaxonomic and phylogenomic evidence, strain USBA 369T is considered to represent a novel genus and a novel species for which the name Consotaella salsifontis gen. nov., sp. nov. is proposed. The type strain is USBA 369T (=KCTC 22549T=CMPUJ U369T).

Hymenobacter gummosus sp. nov., isolated from a spring.

Strain ANT-18T, isolated from a water sample taken from the Longtoushan Spring in Taiwan, was characterized using a polyphasic taxonomic approach. Cells of strain ANT-18T were Gram-stain-negative, aerobic, motile by gliding rods that were surrounded by a thick capsule and it formed pink colonies. Growth occurred at 20-37 °C (optimum, 25 °C), at pH 6-8 (optimum, pH 7) and with 0-1 % NaCl (w/; optimum, 0 %). Phylogenetic analyses, based on 16S rRNA gene sequences, showed that strain ANT-18T belongs to the genus Hymenobacter and showed the highest levels of sequence similarity to Hymenobacter ocellatus Myx 2105T (97.8 %) and Hymenobacter paludis KBP-30T (97.7 %). Strain ANT-18T contained iso-C15 : 0, anteiso-C15 : 0, C18 : 0 and summed feature 4 (iso-C17 : 1 I and/or anteiso-C17 : 1 B) as the predominant fatty acids. The major isoprenoid quinone was MK-7. The polar lipid profile consisted of phosphatidylethanolamine, one uncharacterized aminolipid, two uncharacterized aminophospholipids, two uncharacterized glycolipids and seven uncharacterized lipids. The major polyamine was homospermidine. The genomic DNA G+C content of strain ANT-18T was 63.6 mol%. The DNA-DNA relatedness values of strain ANT-18T with respect to H. ocellatus Myx 2105T and H. paludis KBP-30T were less than 35 %. On the basis of the phylogenetic inference and phenotypic data, strain ANT-18T is recognized to be a representative of a novel species within the genus Hymenobacter. The name Hymenobacter gummosus sp. nov. is proposed, with strain ANT-18T (=BCRC 80966T=LMG 29477T=KCTC 52166T) as the type strain.

Desulfuribacillus stibiiarsenatis sp. nov., an obligately anaerobic, dissimilatory antimonate- and arsenate-reducing bacterium isolated from anoxic sediments, and emended description of the genus Desulfuribacillus.

A novel anaerobic, Gram-stain-negative, endospore-forming bacterium, designated strain MLFW-2T, was isolated from anoxic sediments collected from the drainage area of a geothermal spring near Mono Lake, CA, USA. Optimal growth was achieved at 34 °C and pH 8.25-8.50 in medium containing 0.75 % (w/v) NaCl. Catalase, but not oxidase, was produced. Strain MLFW-2T was an obligate anaerobe capable of respiring with nitrate, nitrite, DMSO, arsenate, antimonate, selenate and selenite as terminal electron acceptors. Lactate, pyruvate, formate and H2 could serve as electron donors to support growth. The isolate was incapable of fermentation. The predominant fatty acids were C16 : 0, C16 : 1ω9c, C16 : 1ω7c, C18 : 1ω9c and C18 : 1ω7c. The major polar lipids were phosphatidylglycerol and phosphatidylethanolamine. The only isoprenoid quinone detected was menaquinone 7 (MK-7). The DNA G+C content was 38.2 mol%. Phylogenetic analysis based on the 16S rRNA gene sequence demonstrated that strain MLFW-2T was a member of the order Bacillales and was most closely related to Desulfuribacillus alkaliarsenatis AHT28T (93.9 % similarity). On the basis of phenotypic and phylogenomic evidence, strain MLFW-2T represents a novel species of the genus Desulfuribacillus, for which the name Desulfuribacillus stibiiarsenatis sp. nov. is proposed. The type strain is MLFW-2T (=DSM 28709T=JCM 30866T). An emended description of the genus Desulfuribacillus is also provided.

Flectobacillus pallidus sp. nov., isolated from a freshwater spring.

A novel bacterial strain, MVW-6T, was isolated from a freshwater spring in Taiwan and characterized using a polyphasic taxonomy approach. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain MVW-6T belonged to the genus Flectobacillus and showed the highest levels of sequence similarity to Flectobacillus lacus CL-GP79T (98.2 % 16S rRNA gene sequence similarity) and Flectobacillus fontis MIB-4T (97.4 %). Cells of strain MVW-6T were Gram-stain-negative, aerobic, non-motile rods that were covered by large capsules and formed light pink colonies. Growth occurred at 15-37 °C (optimum 15-25 °C), at pH 5-9 (optimum pH 7) and with 0-0.5 % (w/v) NaCl (optimum 0 %). Strain MVW-6T contained iso-C15 : 0, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), C16 : 1ω5c and C16 : 0 as the predominant fatty acids. The polar lipid profile consisted of phosphatidylethanolamine, five unidentified aminophospholipids, one unidentified glycolipid, one unidentified phospholipid, one unidentified aminolipid and two unidentified lipids. The major isoprenoid quinone was MK-7. The G+C content of the genomic DNA was 39.5 mol%. DNA-DNA hybridization values for strain MVW-6T with Flectobacillus lacus CL-GP79T and Flectobacillus fontis MIB-4T were less than 50 %. On the basis of the phylogenetic inference and phenotypic data, strain MVW-6T a novel species of the genus Flectobacillus, for which the name Flectobacillus pallidus sp. nov. is proposed. The type strain is MVW-6T (=BCRC 80975T=LMG 29555T=KCTC 33800T).

Flectobacillus fontis sp. nov., isolated from a freshwater spring.

A bacterial strain, designated MIB-4T, was isolated from a freshwater spring in Taiwan and characterized using a polyphasic taxonomy approach. Cells of strain MIB-4T were Gram-staining-negative, aerobic, non-motile rods that formed pale pink colonies. Growth occurred at 15-30 °C (optimum, 25 °C), at pH 7-8 (optimum, pH 7) and with 0-0.5 % NaCl (optimum, 0 %). Phylogenetic analyses based on 16S rRNA gene sequences showed that strain MIB-4T belonged to the genus Flectobacillus and was most closely related to Flectobacillus lacus CL-GP79T with sequence similarity of 98.7 %. Strain MIB-4T contained C16 : 1ω5c, iso-C15 : 0 and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) as the predominant fatty acids. The major isoprenoid quinone was MK-7. The polar lipid profile consisted of phosphatidylethanolamine, six unidentified aminophospholipids, one unidentified glycolipid, one unidentified phospholipid and three unidentified lipids. The DNA G+C content of the genomic DNA was 37.6 mol%. The DNA-DNA hybridization value for strain MIB-4T with F. lacus CL-GP79T was less than 32 %. On the basis of the phylogenetic inference and phenotypic data, strain MIB-4T should be classified as a representative of a novel species, for which the name Flectobacillus fontis sp. nov. is proposed. The type strain is MIB-4T (=BCRC 80926T=LMG 29292T=KCTC 33763T).

Soortia roseihalophila gen. nov., sp. nov., a new taxon in the order Balneolales isolated from a travertine spring, and description of Soortiaceae fam. nov.

A novel Gram-stain-negative, slightly halophilic, motile, curved rod with a horseshoe shape, designated strain Bsw-2bT, was isolated from Badab-Soort travertine spring in Iran. Phylogenetic analysis of the 16S rRNA gene sequence revealed that strain Bsw-2bT belongs to the order Balneolales, showing 84.6 % sequence similarity to Gracilimonastropica DSM 19535T and 84.4 % and 83.9 % sequence similarity to Gracilimonas rosea CL-KR2T and Balneola vulgaris DSM 17893T, respectively. In addition, phenotypic and physiological features could clearly differentiate strain Bsw-2bT from species of the most closely related genera, Gracilimonas, Balneola, Aliifodinibius and Fodinibius. The strain was able to grow with 1-3 % (w/v) (optimum at 2 %) NaCl, at temperatures of 28-34 °C (optimum at 30 °C) and between pH 6.0 and 8.0 (optimum at pH 7.0). The major cellular fatty acids of strain Bsw-2bT were iso-C15 : 0, iso-C13 : 0 and iso-C14 : 0. The polar lipid profile of strain Bsw-2bT was composed predominantly of phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, an unknown glycolipid and four unknown phospholipids. The DNA G+C content was 40.5 mol%. Based on the evidence from the polyphasic study, strain Bsw-2bT represents a novel species in a novel genus within a new family, for which the name Soortia roseihalophila gen. nov., sp. nov. is proposed, within the new family Soortiaceae fam. nov. The type strain is strain Bsw-2bT (=IBRC-M 10915T=LMG 28547T).

Actinocrinis puniceicyclus gen. nov., sp. nov., an actinobacterium isolated from an acidic spring.

An aerobic, mildly acidophilic actinobacterium was isolated from the Ochre Beds bog in Kootenay National Park, Canada. Cells of isolate OB1T were Gram-stain-positive, non-motile, pink- to purple-pigmented filaments. The pH range for growth was pH 3.5-6.5 (optimum pH 5.5), and the temperature range was 13-30°C. The major cellular fatty acids were i-C16 : 0 (28.5 %), i-C15 : 0 (14.6 %) and ai-C15 : 0 (14.3 %), and the major polar lipid was phosphohexose. The major quinone was menaquinone-11 (MK-11), and the peptidoglycan type was A1γ. The DNA G+C content was 70.2 %. Along with growth on complex media including yeast extract, proteose peptone, casamino acids and tryptic soy broth, growth occured on mono- and disaccharides (glucose, sucrose, galactose and xylose) and polysaccharides (starch, gellan, pectin, xylan and alginate). Anaerobic growth was not observed. The cells did not fix atmospheric nitrogen. On the basis of comparative 16S rRNA gene sequence analysis, this isolate belonged to the family Actinospicaceae, in the suborder Catenulisporineae of the order Actinomycetales. The most closely related species was Actinospica robiniae. However, the 16S rRNA gene sequence identity to this bacterium was low (92.8 %) and there were several chemotaxonomic differences from this species. We therefore propose a novel genus and species, Actinocrinis puniceicyclus gen. nov., sp. nov., with strain OB1T (=DSM 45618T=ATCC BAA-2771T) as the type strain.

Flavobacterium dispersum sp. nov., isolated from a freshwater spring.

A novel bacterial strain MVW-23T was isolated from a freshwater spring in Taiwan. The strain was Gram-staining-negative, strictly aerobic, motile by gliding, rod-shaped and formed translucent yellow colonies. Optimal growth occurred at 20-30 °C, pH 7.0, and in the presence of 0.5-1 % NaCl. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain MVW-23T belonged to the genus Flavobacterium and showed the highest levels of sequence similarity with respect to Flavobacterium denitrificans ED5T (97.3 %), Flavobacterium kyungheense THG-107T (97.2 %) and Flavobacterium defluvii EMB117T (97.0 %). Strain MVW-23T contained iso-C15 : 0, summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c) and iso-C17 : 0 3-OH as the predominant fatty acids. The polar lipid profile consisted of phosphatidylethanolamine, three uncharacterized aminophospholipids and one uncharacterized phospholipid. The major polyamine was homospermidine. The major isoprenoid quinone was MK-6. The DNA G+C content of the genomic DNA was 39.9 mol%. The DNA-DNA hybridization value for strain MVW-23T with F. denitrificans DSM 15936T, F. kyungheense LMG 26575T and F. defluvii DSM 17963T was less than 35 %. Differential phenotypic properties, together with the phylogenetic inference, demonstrate that strain MVW-23T should be classified as a novel species of the genus Flavobacterium, for which the name Flavobacterium dispersum sp. nov. is presented. The type strain is MVW-23T (=BCRC 80978T=LMG 29558T=KCTC 52234T).

Novosphingobium fontis sp. nov., isolated from a spring.

A bacterial strain designated STM-14T was isolated from a spring in Taiwan and characterized using a polyphasic taxonomic approach. Strain STM-14T was a Gram-stain-negative, aerobic, poly-ß-hydroxybutyrate-accumulating, non-motile, rod-shaped bacterium and formed cream-coloured colonies. Strain STM-14T grew at 15-37 °C (optimum, 30 °C), at pH 5.0-9.0 (optimum, pH 6.0) and with 0-0.5 % (w/v) NaCl (optimum, 0 %). Phylogenetic analyses based on 16S rRNA gene sequences showed that strain STM-14T belonged to the genus Novosphingobium and showed the highest levels of sequence similarity to Novosphingobium naphthae D39T (98.5 %) and Novosphingobium mathurense SM117T (97.6 %). The major fatty acids (>10 %) of strain STM-14T were C17 : 1ω6c, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), C18 : 1ω7c, C15 : 0 2-OH and C17 : 1ω8c. The polar lipid profile consisted of phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol, phosphatidyldimethylethanolamine, sphingoglycolipid, phosphatidylcholine and several uncharacterized lipids. The major polyamine was spermidine. The major isoprenoid quinone was Q-10. The DNA G+C content was 66.6 mol%. The DNA-DNA hybridization value for strain STM-14T with N. naphthae JCM 31158T and N. mathurense DSM 23374T was less than 49 %. Differential phenotypic properties, together with the phylogenetic inference, demonstrate that strain STM-14T should be classified as representing a novel species of the genus Novosphingobium, for which the name Novosphingobium fontis sp. nov. is proposed. The type strain is STM-14T (=BCRC 80924T=LMG 29290T=KCTC 42983T).

Chitinibacter fontanus sp. nov., isolated from a spring.

A bacterial strain, designated STM-7T, was isolated from a spring in Taiwan and characterized using a polyphasic taxonomy approach. Cells of strain STM-7T were Gram-staining-negative, aerobic, poly-ß-hydroxybutyrate-accumulating, motile by a single polar flagellum, rod-shaped, surrounded by a thick capsule and formed milky-white colonies. Growth occurred at 15-37 °C (optimum, 25-30 °C), at pH 6-8 (optimum, pH 6-7) and with 0-2 % NaCl (optimum, 0-1 %). Phylogenetic analyses based on 16S rRNA gene sequences showed that strain STM-7T belonged to the genus Chitinibacter and was most closely related to Chitinibacter tainanensis S1T with a sequence similarity of 97.3 %. Strain STM-7T contained summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c) and C16 : 0 as the predominant fatty acids. The major hydroxyl fatty acids were C12 : 0 3-OH and C16 : 0 3-OH. The polar lipid profile consisted of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, an uncharacterized aminophospholipid, an uncharacterized glycolipid and an uncharacterized phospholipid. The major isoprenoid quinone was Q-8. The DNA G+C content of the genomic DNA was 52.4 mol%. The DNA-DNA hybridization value for strain STM-7T with Chitinibacter tainanensis BCRC 17254T was less than 47 %. On the basis of the phylogenetic inference and phenotypic data, strain STM-7T should be classified as a representative of a novel species, for which the name Chitinibacter fontanus sp. nov. is proposed. The type strain is STM-7T (=BCRC 80923T=LMG 29289T=KCTC 42982T).