Comparative genome sequence analysis of several species in the genus Tepidimonas and the description of a novel species Tepidimonas charontis sp. nov.

We performed high-quality genome sequencing of eight strains of the species of the genus Tepidimonas and examined the genomes of closely related strains from the databases to understand why Tepidimonas taiwanensis is the only strain of this genus that utilizes glucose and fructose for growth. We found that the assimilation of these hexoses by T. taiwanensis was due to the presence of two transporters that are absent in all other genomes of strains of members of the genus Tepidimonas examined. Some strains lack genes coding for glucokinase, but the Embden-Meyerhof-Parnas pathway appears to be otherwise complete. The pentose phosphate pathway has a complete set of genes, but genes of the Entner-Doudoroff pathway were not identified in the genomes of any of the strains examined. Genome analysis using average nucleotide identity (ANIb), digital DNA-DNA hybridization (dDDH), average amino acid identity (AAI) and phylogenetic analysis of 400 conserved genes was performed to assess the taxonomic classification of the organisms. Two isolates of the genus Tepidimonas from the hot spring at São Pedro do Sul, Portugal, designated SPSP-6T and SPSPC-18 were also examined in this study. These organisms are mixotrophic, have an optimum growth temperature of about 50 ºC, utilize several organic acids and amino acids for growth but do not grow on sugars. Distinctive phenotypic, 16S rRNA gene sequence and genomic characteristics of strains SPSP-6T and SPSPC-18 lead us to propose a novel species based on strain SPSP-6T for which we recommend the name Tepidimonas charontis sp. nov. (=CECT 9683T=LMG 30884T).

Transfer of Meiothermus chliarophilus (Tenreiro et al.1995) Nobre et al. 1996, Meiothermus roseus Ming et al. 2016, Meiothermus terrae Yu et al. 2014 and Meiothermus timidus Pires et al. 2005, to Calidithermus gen. nov., as Calidithermus chliarophilus comb. nov., Calidithermus roseus comb. nov., Calidithermus terrae comb. nov. and Calidithermus timidus comb. nov., respectively, and emended description of the genus Meiothermus.

Chemotaxonomic parameters, phylogenetic analysis of the 16S rRNA gene, phylogenetic analysis of 90 housekeeping genes and 855 core genes, amino acid identity (AAI), average nucleotide identity (ANI) and genomic characteristics were used to examine the 13 species of the genus Meiothermuswith validly published names to reclassify this genus. The results indicate that the species of the genus Meiothermus can be divided into three lineages on the basis of the results of the phylogenetic analysis, AAI, the guanine+cytosine (G+C) mole ratio, the ability to synthesize the red-pigmented carotenoid canthaxanthin and the colony colour, as well as other genomic characteristics. The results presented in this study circumscribe the genus Meiothermus to the species Meithermus ruber, Meiothermus cateniformans, Meiothermus taiwanensis, Meiothermus cerbereus, Meiothermus hypogaeus, Meiothermus luteus, Meiothermus rufus and Meiothermus granaticius, for which it is necessary to emend the genus Meiothermus. The species Meiothermus silvanus, which clearly represents a separate genus level lineage was not reclassified in this study for lack of any distinctive phenotypic or genotypic characteristics. The results of this study led us to reclassify the species Meiothermus chliarophilus, Meiothermus timidus, Meiothermus roseus and Meiothermus terrae as species of a novel genus for which we propose the epithet Calidithermus gen. nov.

OSTEODENSITOMETRY AND TOMOGRAPHIC FINDINGS IN FOUR CAPTIVE GIANT SOUTH AMERICAN TURTLES (PODOCNEMIS EXPANSA) WITH METABOLIC BONE DISEASE.

Giant South American turtles (Podocnemis expansa) are at a risk of extinction because of the rapid decline in their population over the last few decades. Metabolic bone disease (MBD) is common in captive testudines, but is often not diagnosed until a later stage. The authors present the cases of four captive giant South American turtles with carapace deformity secondary to MBD that underwent computed tomography (CT) scans of the carapace bones and vertebral column. Findings indicative of changes in geometry were found in both. The cancellous bone pattern was characterized by varying degrees of increased trabecular spacing and cortical thinning of the pleural bones. Bone densitometry analysis of the pleural and neural bones and at the level of the body of the third, fourth, and fifth dorsal vertebrae showed mean density values much lower than those found in two adult specimens of the same species that were considered healthy. In conclusion, CT contributed important information on the degree of demineralization and possible structural changes due to MBD and should be considered a relevant tool for diagnosis of this condition.

Characterization of amputees at a large hospital in Recife, PE, Brazil.

BACKGROUND: Limb amputation can be defined as a procedure that consists of separating a limb or a segment of a limb from the body. OBJECTIVES: To describe the profile of limb amputation procedures performed at a large hospital run by the state of Pernambuco (Brazil). METHODS: Cross-sectional descriptive and retrospective study conducted at a large hospital in the city of Recife, PE. Data were collected from the records of patients who underwent amputations during 2017. Records from patients who had had a limb amputation during 2017 were included, unless data were illegible or missing. RESULTS: A total of 328 procedures were performed on 274 patients, the majority of whom were male (57.7%). There was a predominance of lower limb amputations (64.2%), of non-traumatic causes (86.5%), and urgent treatment (96.4%). The majority of patients who underwent amputations remained in hospital for 11 to 25 days (32.1%). The study found that the majority of amputees were discharged (69.7%), although a proportion died. Deaths of lower limb amputees were primarily among elderly women in the age range of 60 to 90 years (76%), females (55%), and patients subjected to a single amputation (91%). CONCLUSIONS: The data observed in this study are alarming, particularly considering that many of these amputations could have been avoided, since they were caused by complications of diseases that can be prevented and controlled at healthcare services of a lower level of complexity and at a relatively low cost.

Lysobacter silvestris sp. nov., isolated from alpine forest soil, and reclassification of Luteimonas tolerans as Lysobacter tolerans comb. nov.

A Gram-stain-negative, rod-shaped, motile, catalase-positive and cytochrome c oxidase-positive bacterial strain, designated AM20-91T, was isolated from alpine forest soil. Phylogenetic analysis based on 16S rRNA gene sequencing showed that strain AM20-91T was related to the genus Lysobacter and had highest 16S rRNA gene sequence similarities to the type strains of Lysobacter novalis THG-PC7T (97.8 %), Luteimonas tolerans UM1T (97.7 %) and Lysobacter ximonensis XM415T (97.0 %). The strain contained ubiquinone 8 as the predominant respiratory quinone; its polar lipid profile contained phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and two unidentified aminophospholipids. The major cellular fatty acids (>10 %) were iso-C15 : 0, iso-C11 : 0 3-OH and iso-C11 : 0. The DNA G+C content was 63.35 % (draft genome sequence). The combined results of phylogenetic, phenotypic, DNA-DNA relatedness and chemotaxonomic analyses demonstrated that strain AM20-91T represents a novel species of the genus Lysobacter, for which the name Lysobacter silvestris sp. nov. is proposed. The type strain is AM20-91T (=DSM 104734T=LMG 30011). In this study, it is also proposed that Luteimonas tolerans be reclassified as member of the genus Lysobacter.

Raineya orbicola gen. nov., sp. nov. a slightly thermophilic bacterium of the phylum Bacteroidetes and the description of Raineyaceae fam. nov.

An isolate, designated SPSPC-11T, with an optimum growth temperature of about 50 °C and an optimum pH for growth between 7.5 and 8.0, was recovered from a hot spring in central Portugal. Based on phylogenetic analysis of its 16S rRNA sequence, the new organism is most closely related to the species of the genus Thermonema but with a pairwise sequence similarity of <85 %. The isolate was orange-pigmented, formed non-motile long filaments and rod-shaped cells that stain Gram-negative. The organism was strictly aerobic, oxidase-positive and catalase-positive. The major fatty acids were iso-C15:0, iso-C15 : 0 2-OH and iso-C17 : 0 3-OH. The major polar lipids were one aminophospholipid, two aminolipids and three unidentified lipids. Menaquinone 7 was the major respiratory quinone. The DNA G+C content of strain SPSPC-11T was 37.6 mol% (draft genome sequence). The high quality draft genome sequence corroborated many of the phenotypic characteristics of strain SPSPC-11T. Based on genotypic, phylogenetic, physiological and biochemical characterization we describe a new species of a novel genus represented by strain SPSPC-11T (=CECT 9012T=LMG 29233T) for which we propose the name Raineya orbicola gen. nov., sp. nov. We also describe the family Raineyaceae to accommodate this new genus and species.

Solimicrobium silvestre gen. nov., sp. nov., isolated from alpine forest soil.

A Gram-stain-negative, rod-shaped, motile, catalase and cytochrome c oxidase-positive bacterial strain, designated S20-91T, was isolated from alpine forest soil. Growth occurred within a temperature range of 0-25 °C. Yeast extract was required for growth. Phylogenetic analysis based on 16S rRNA gene sequencing showed that strain S20-91T was related to the genus Herminiimonas and had the highest 16S rRNA gene sequence similarity to Herminiimonas arsenicoxydans ULPAs1T (96.5 %). The strain contained ubiquinone 8 as the predominant respiratory quinone and phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol as the major polar lipids. The major cellular fatty acids (>10 %) were C16 : 1ω7c (55.3 %) and C16 : 0 (25.6 %). The genomic DNA G+C content was 47.6 mol%. Combined data of genomic, phylogenetic, phenotypic and chemotaxonomic analyses demonstrated that strain S20-91T represents a novel genus and species, for which the name Solimicrobium silvestre gen. nov., sp. nov. is proposed. The type strain is S20-91T (=DSM 104733T=LMG 30010).

Mariniblastus fucicola gen. nov., sp. nov. a novel planctomycete associated with macroalgae.

One strain of a novel genus and species of the order Planctomycetes, designated FC18T, was isolated from the epiphytic community of Fucusspiralis. This strain was non-pigmented in medium M13 but was slightly pink pigmented on medium M14, containing four-fold the levels of glucose, peptone and yeast extract of M13. The organism was primarily spherical, with unicellular non-motile forms and rosettes. The optimal temperature for growth was about 25 °C and the optimal pH was 7.5. FC18T was chemoorganotrophic and aerobic. Several sugars, polyols and amino acids were assimilated. The major fatty acids were C18 : 1ω9c, C14 : 0 and C16 : 0. The major polar lipids were phosphatidylglycerol (PG) and two unknown lipids. Menaquinone 5 (MK-5) was the main respiratory quinone, but MK-6 was also present. The results of the 16S rRNA gene sequence analysis confirmed the affiliation of this organism to the order Planctomycetales, family Planctomycetaceae, with Blastopirellula marina as the closest relative with only 86 % sequence similarity. On the basis of physiological, biochemical and chemotaxonomic characteristics we propose that FC18T(=LMG 29748T=DSM 26290T) represents a novel species of a novel genus of the family Planctomycetaceae for which we propose the name Mariniblastusfucicola gen. nov., sp. nov.

Psychromicrobium silvestre gen. nov., sp. nov., an actinobacterium isolated from alpine forest soils.

Two Gram-stain-variable, non-motile, catalase-positive and cytochrome c oxidase-negative bacteria, designated AK20-18T and AM20-54, were isolated from forest soil samples collected in the Italian Alps. Growth occurred at a temperature range of 5-30 °C, at pH 6-9 and in the presence of 0-5 % (w/v) NaCl. The 16S rRNA gene sequence similarity between strains AK20-18T and AM20-54 was 100 %. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain AK20-18T had highest 16S rRNA gene sequence similarity with the type strain of Arthrobacter psychrochitiniphilus (96.9 %). The cell-wall peptidoglycan structure of strain AK20-18T was of the type A3alpha l-Lys-l-Thr-l-Ala2 (A11.27). The whole-cell sugars were galactose, ribose and lesser amounts of mannose. The major respiratory quinone of the two strains was menaquinone 9(H2) [MK-9(H2)], whereas MK-10(H2) was a minor component. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and unknown glycolipids. The major cellular fatty acids were anteiso-C15 : 0, iso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0. The genomic DNA G+C content was 59.9 mol%. Combined data of phylogenetic, phenotypic and chemotaxonomic analyses demonstrated that strains AK20-18T and AM20-54 represent a novel genus and species, for which the name Psychromicrobium silvestre gen. nov., sp. nov. is proposed. The type strain of Psychromicrobium silvestregen. nov., sp. nov. is AK20-18T (=DSM 102047T=LMG 29369T).

Dehalogenimonas formicexedens sp. nov., a chlorinated alkane-respiring bacterium isolated from contaminated groundwater.

A strictly anaerobic, Gram-stain-negative, non-spore-forming bacterium designated NSZ-14T, isolated from contaminated groundwater in Louisiana (USA), was characterized using a polyphasic approach. Strain NSZ-14T reductively dehalogenated a variety of polychlorinated aliphatic alkanes, producing ethene from 1,2-dichloroethane, propene from 1,2-dichloropropane, a mixture of cis- and trans-1,2-dichloroethene from 1,1,2,2-tetrachloroethane, vinyl chloride from 1,1,2-trichloroethane and allyl chloride (3-chloro-1-propene) from 1,2,3-trichloropropane. Formate or hydrogen could both serve as electron donors. Dechlorination occurred between pH 5.5 and 7.5 and over a temperature range of 20-37 °C. Major cellular fatty acids included C18 : 1ω9c, C14 : 0 and C16 : 0. 16S rRNA gene sequence-based phylogenetic analysis indicated that the strain clusters within the class Dehalococcoidia of the phylum Chloroflexi, most closely related to but distinct from type strains of the species Dehalogenimonas alkenigignens (97.63 % similarity) and Dehalogenimonas lykanthroporepellens (95.05 %). A complete genome sequence determined for strain NSZ-14T revealed a DNA G+C content of 53.96 mol%, which was corroborated by HPLC (54.1±0.2 mol% G+C). Genome-wide comparisons based on average nucleotide identity by orthology and estimated DNA-DNA hybridization values combined with phenotypic and chemotaxonomic traits and phylogenetic analysis indicate that strain NSZ-14T represents a novel species within the genus Dehalogenimonas, for which the name Dehalogenimonas formicexedens sp. nov. is proposed. The type strain is NSZ-14T (=HAMBI 3672T=JCM 19277T=VKM B-3058T). An emended description of Dehalogenimonas alkenigignens is also provided.

Ampullimonas aquatilis gen. nov., sp. nov. isolated from bottled mineral water.

Two isolates, designated B15.09-116T and B15.09-124, were recovered from bottled mineral water in Portugal. Based on 16S rRNA gene sequence analysis, these strains were related most closely to species of the genus Derxia (belonging to the family Alcaligenaceae) with pairwise sequence similarities of 93.0-93.6 %. The isolates were not pigmented and formed Gram-stain-negative, short, motile rod-shaped cells. The organisms were strictly aerobic, oxidase-positive and catalase-negative. These organisms also fixed N2. The major polar lipids were phosphatidylglycerol and phosphatidylethanolamine. Ubiquinone 8 was the major respiratory quinone. The DNA G+C content of strain B15.09-116T was 49.8 mol%. Based on phylogenetic, physiological and biochemical characteristics the two strains are considered to represent a novel species of a new genus, for which the name Ampullimonas aquatilis gen. nov., sp. nov. is proposed. The type strain of Ampullimonas aquatilis is B15.09-116T ( = CECT 8581T = LMG 28208T).

Nakamurella silvestris sp. nov., an actinobacterium isolated from alpine forest soil.

A Gram-stain-positive, non-motile, catalase-positive and cytochrome c oxidase-negative bacterium, designated strain S20-107T, was isolated from alpine forest soil. Growth occurred at 0-30 °C, at pH 6-9 and in the presence of 0-3 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain S20-107T was related to the genus Nakamurella and had the highest 16S rRNA gene sequence similarity with Nakamurella flavida DS-52T (96.1 %). Strain S20-107T showed <96.1% 16S rRNA gene sequence similarities to all other recognized members of the genus Nakamurella. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The major whole-cell sugars were glucose, galactose, mannose, arabinose, ribose and rhamnose. The strain contained MK-8(H4) as the predominant menaquinone and diphosphatidylglycerol, phosphatidylethanolamine and an unidentified aminophospholipid as the major polar lipids. The major cellular fatty acids were anteiso-C15 : 0, C16 : 0, summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c and/or iso-C15 : 0 2-OH) and iso-C16 : 0. The genomic DNA G+C content was 70.5 mol%. Combined data of phylogenetic, phenotypic and chemotaxonomic analyses demonstrated that strain S20-107T represents a novel species of the genus Nakamurella, for which the name Nakamurella silvestris sp. nov. is proposed. The type strain is S20-107T (=DSM 102309T=LMG 29427T).

Cavicella subterranea gen. nov., sp. nov., isolated from a deep mineral-water aquifer, and emended description of the species Perlucidibaca piscinae.

One strain designated W2.09-231T was isolated from an aquifer through a 150-metre-deep borehole feeding a mineral-water bottling plant in Central Portugal. Based on 16S rRNA gene sequence analysis, the novel organism is most closely related to the species of the genera Perlucidibaca and Paraperlucidibaca, belonging to the family Moraxellaceae, with 16S rRNA gene pairwise sequence similarity of 94.5 and 93.1 %, respectively. The strain was not pigmented and formed Gram-stain-negative, non-motile, short rod-shaped cells. The organism was strictly aerobic, and oxidase- and catalase-positive. Strain W2.09-231T was organotrophic, but grew only on a very limited number of single carbon sources. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and one major unknown phospholipid. Ubiquinone 12 (U-12) was the major respiratory quinone. The DNA G+C content of strain W2.09-231T was 62.0 mol%. Based on phylogenetic, physiological and biochemical characteristics, we describe a novel species of a novel genus represented by strain W2.09-231T ( = CECT 8582T = LMG 28332T) for which we propose the name Cavicella subterranea gen. nov., sp. nov. We also propose to emend the description of the species Perlucidibaca piscinae to reflect new results obtained in this study.

Palleronia abyssalis sp. nov., isolated from the deep Mediterranean Sea and the emended description of the genus Palleronia and of the species Palleronia marisminoris.

Three strains designated 221-F1(T), 221-F2 and 3030-F1 were isolated from the Matapan Vavilov Deep canyon, also known as Calypso Deep in the Eastern Mediterranean Sea, at a depth of 4,908 m. Based on 16S rRNA gene sequence analysis these strains were found to be most closely related to Palleronia marisminoris and Hwanghaeicola aestuarii, with 16S rRNA gene pairwise sequence similarity of 95.3 and 94.7 % respectively, belonging to the family Rhodobacteraceae. The strains were observed to be red-pigmented and to form non-motile cocci or pleomorphic cells. The cells were found to stain Gram-negative, to be strictly aerobic, oxidase and catalase positive. Strains 221-F1(T), 221-F2 and 3030-F1 were found to be mesophilic and to grow in medium containing up to 13 % NaCl. The major polar lipids of the three strains were identified as diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, an unidentified glycolipid and an unidentified aminolipid. Ubiquinone 10 (U-10) was found to be the major respiratory quinone. The DNA G+C content of strain 221-F1(T) was determined to be 64.7 mol%. Based on phylogenetic, physiological and biochemical characteristics we describe a new species represented by strain 221-F1(T) (=CECT 8504(T) = LMG 27977(T)) for which we propose the name Palleronia abyssalis sp. nov. We also propose to emend the description of the genus Palleronia and the species P. marisminoris to reflect new results obtained in this study.

Thermophiles as potential source of novel endotoxin antagonists: the full structure and bioactivity of the lipo-oligosaccharide from Thermomonas hydrothermalis.

Thermomonas hydrothermalis is a Gram-negative thermophilic bacterium that is able to live at 50 °C. This ability is attributed to chemical modifications, involving those to bacterial cell-wall components, such as proteins and (glyco)lipids. As the main component of the outer membrane of Gram-negative bacteria, lipopolysaccharides (LPSs) are exposed to the environment, thus they can undergo structural chemical changes to allow thermophilic bacteria to live at their optimal growth temperature. Furthermore, as one of the major target of the eukaryotic innate immune system, LPS elicits host immune response in a structure-dependent mode; thus the uncommon chemical features of thermophilic bacterial LPSs might exert a different biological action on the innate immune system-an antagonistic effect, as shown in studies of LPS structure-activity relationship in the ongoing research into antagonist LPS candidates. Here, we report the complete structural and biological activity analysis of the lipo-oligosaccharide isolated from Thermomonas hydrothermalis, achieved by a multidisciplinary approach (chemical analysis, NMR, MALDI MS and cellular immunology). We demonstrate a tricky and interesting structure combined with a very interesting effect on human innate immunity.

Cecembia calidifontis sp. nov., isolated from a hot spring runoff, and emended description of the genus Cecembia.

Two red-pigmented isolates, designated RQ-33(T) and TU-13, were recovered from hot spring runoffs on the Island of São Miguel in the Azores, Portugal. These organisms have an optimum growth temperature of approximately 45 °C and an optimum pH for growth between 7.5 and 8.5. Based on 16S rRNA gene sequence analysis, the novel isolates were most closely related to the type strain of Cecembia lonarensis at 96.4 % pairwise similarity. Cells of the two isolates were non-motile, rod-shaped, Gram-stain-negative, strictly aerobic, and oxidase- and catalase-positive. The major fatty acids were iso-C15 : 0 and C15 : 0, phosphatidylethanolamine was the major polar lipid, and menaquinone 7 was the major respiratory quinone. Based on phylogenetic analyses, and physiological and biochemical characteristics, these isolates from the Azores represent a novel species of the genus Cecembia for which the name Cecembia calidifontis is proposed with strain RQ-33(T) ( = DSM 21411(T) = LMG 24596(T)) as the type strain. The description of the genus Cecembia is also emended.

Pullulanibacillus uraniitolerans sp. nov., an acidophilic, U(VI)-resistant species isolated from an acid uranium mill tailing effluent and emended description of the genus Pullulanibacillus.

Two Gram-positive-staining, rod-shaped, endospore-forming isolates (UG-2(T) and UG-3), with an optimum growth temperature of around 37 °C and an optimum pH for growth of about 4, were recovered from an acidic effluent of the uranium mill tailing at Urgeiriça in Central Portugal. On the basis of 16S rRNA gene sequence similarity, the strains belonged to the family Sporolactobacillaceae and were closely related to Pullulanibacillus naganoensis ATCC 53909(T) (97.9 %). Unlike P. naganoensis, strains UG-2(T) and UG-3 grew in medium containing up to 5000 p.p.m. U(VI) but did not hydrolyse pullulan. Chemotaxonomic data also supported the affiliation of strains UG-2(T) and UG-3 to the genus Pullulanibacillus. Physiological and biochemical tests along with fatty acid composition allowed differentiation of strains UG-2(T) and UG-3 from P. naganoensis. It is suggested that strains UG-2(T) and UG-3 represent a novel species, for which the name Pullulanibacillus uraniitolerans is proposed; the type strain is UG-2(T) (=DSM 19429(T) = LMG 24205(T)). An emended description of the genus Pullulanibacillus is also proposed.