Archives of Microbiology

Microbacterium fakhimi sp. nov. has been isolated from a contaminated algal culture (Chlamydomonas reinhardtii). Its genome has been fully sequenced (3,753,259 base pairs) and a tentative annotation is provided (3,704 genes). Both, genome information and growth tests suggest that M. fakhimi sp. nov. is auxotroph for biotin and thiamine and unable to use sulfate as sulfur (S) source. S-reduced forms, such as methionine and cysteine can support M. fakhimi sp. nov. growth. The potential biotechnological interest of this bacteria is discussed here and in a related research paper (Fakhimi et al., 2023b).

Following the announcement of SARS-CoV-2 worldwide pandemic spread by WHO on March 11, 2020, wastewater based epidemiology received great attention in several countries: The Netherlands [Medama et al., 2020; K-Lodder et al., 2020], USA [Wu et al., 2020; Memudryi et al., 2020], Australia [Ahmed et al., 2020], France [Wurtzer et al., 2020], China [Wang et al., 2020], Spain [Randazzo et al., 2020; Walter et al., 2020], Italy (La Rosa et al., 2020; Rimoldi et al., 2020) and Israel [Or et al., 2020], performed analysis in wastewaters by using different virus concentration techniques. Turkey took its place among these countries on 7th of May, 2020 by reporting SARS-CoV-2 RT-qPCR levels at the inlet of seven (7) major municipal wastewater treatment plants (WWTPs) of Istanbul [Alpaslan Kocamemi et al., 2020], which is a metropole with 15.5 million inhabitants and a very high population density (2987 persons/km2) and having about 65 % of Covid-19 cases in Turkey. Sludges that are produced in WWTPs should be expected to contain SARS-CoV-2 virus as well. There has not yet been any study for the fate of SAR-CoV-2 in sludges generated from WWTPs. Knowledge about the existing of SARS-CoV-2 in sludge may be useful for handling the sludge during its dewatering, stabilizing and disposal processes. This information will also be valuable in case of sludges that are used as soil conditioners in agriculture or sent to landfill disposal. In wastewater treatment plants, generally two different types of sludges are generated; primary sludge (PS) and waste activated sludge (WAS). PS forms during the settling of wastewater by gravity in the primary settling tanks. Little decomposition occurs during primary sludge formation. Since most of the inorganic part of the wastewater is removed in the earlier grit removal process, the PS consists of mainly organic material that settles. The PS is about 1-2 % solids by weight. In the biological treatment part of the WWTPs, the biomass that forms in the anaerobic, anoxic and oxic zones of the process is settled in final clarifiers by gravity and returned to the beginning of the biological process so that it is not washed off. The waste activated sludge (WAS) is the excess part of the biomass that grows in this secondary treatment process. It has to be removed from the process not to increase the mixed liquor suspended solids concentration (bacteria concentration) in the secondary process more than a fixed value. The WAS is about 0.6 - 0.9 % solids by weight. This work aims to find whether SARS-CoV-19 is present in the PS and WAS before it is dewatered and sent to anaerobic or aerobic digester processes or to thermal drying operations. For this purpose, on the 7th of May 2020, two (2) PS samples were collected from Ambarli and Tuzla WWTPs, seven (7) WAS samples were collected from Terkos, Ambarli, Atakoy I & II, Pasakoy II, Buyukcekmece and Tuzla I WWTPs. Polyethylene glycol 8000 (PEG 8000) adsorption [Wu et al., 2020] SARS-Cov-2 concentration method was used for SARS-CoV-2 concentration after optimization. [Alpaslan Kocamemi et al., 2020]. Real time RT-PCR diagnostic panel validated by US was used to quantify SARS-CoV-2 RNA in primary and waste activated sludge samples taken from WWTPs in Istanbul. All samples were tested positive. Titers of SARS-CoV-2 have been detected ranging copies between 1.17x104 to 4.02x104 per liter. O_LSTValue of the DataC_LSTO_LIThe dataset provides information about SARS-CoV-19 in primary and waste activated sludges generated in WWTPs. C_LIO_LIAs being the first study in the world, the dataset presented is expected to be beneficial in handling the sludge during its dewatering, stabilizing and disposal processes C_LI Data DescriptionSARS-CoV-2 copy numbers per liter measured for sludge samples from WWTPs were summarized in Table 1 and shown in Figure 1 together with SARS-CoV-2 copy numbers observed in an earlier study [Alpaslan Kocamemi et al., 2020] in the influent of the WWTPs from which the sludge samples were taken. O_TBL View this table: org.highwire.dtl.DTLVardef@1647aa7org.highwire.dtl.DTLVardef@1b09df2org.highwire.dtl.DTLVardef@518f39org.highwire.dtl.DTLVardef@92061forg.highwire.dtl.DTLVardef@cfe4b2_HPS_FORMAT_FIGEXP M_TBL O_FLOATNOTable 1.C_FLOATNO O_TABLECAPTIONSARS-CoV-2 RT-qPCR results of sludges taken from Istanbul WWTPs C_TABLECAPTION C_TBL O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=129 SRC="FIGDIR/small/20099358v1_fig1.gif" ALT="Figure 1"> View larger version (18K): org.highwire.dtl.DTLVardef@af6ccaorg.highwire.dtl.DTLVardef@10f7150org.highwire.dtl.DTLVardef@d83bbborg.highwire.dtl.DTLVardef@3980c8_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFigure 1.C_FLOATNO SAR-CoV-2 Levels in primary and waste activated sludges of Istanbul WWTPs. C_FIG To the best of our knowledge, no study has yet been reported the presence of SARS-CoV-2 in primary sludge (PS) and waste activated sludge (WAS) samples. Herein we report the results of SARS-CoV-2 presence in two (2) PS and seven (7) WAS samples from WWTPs in Istanbul. A total of nine (9) sludge samples were collected on the 7th May of 2020 and investigated for presence of SARS-CoV-2 with RT-qPCR methodology. SARS-CoV-2 genome was detected quantitatively from all samples. Sludge samples presented CT ranging from 33.5 to 35.8. Titers of SARS-CoV-2 have been detected ranging from 1.17x104 to 4.02x104 per liter. The detected numbers of SARS-CoV-2 in PS samples were found similar to those observed for WAS samples. SARS-CoV-2 copy numbers detected in PS and WAS on 7th of May, 2020 are greater than the copy numbers observed in the influent of these WWTPs on 21st April, 2020 [Alpaslan Kocamemi, 2020]. By considering the fact that the number of cases reported for Istanbul on the 7th of May, 2020 is less than the cases reported for the 21st April, 2020, it may be concluded that SARS-CoV-2 concentrations are more in both primary and waste activated sludge.

Stenotrophomonas goyi sp. nov. has been isolated from a contaminated algal culture (Chlamydomonas reinhardtii). Its genome has been fully sequenced (4,487,389 base pairs) and a tentative annotation is provided (4,147 genes). The genome information suggests that S. goyi sp. nov. is unable to use sulfate and nitrate as sulfur and nitrogen sources, respectively. Growth tests have confirmed the dependence of the sulfur-containing amino acids methionine and cysteine. The potential biotechnological interest of this bacteria is discussed here and in a related research paper (Fakhimi et al., 2023b).

We isolated a gammaproteobacterial methanotrophic strain FWC3, from canal sediment from Western India. The strain oxidizes methane and can also grow on methanol. The draft genome of the same was sequenced which showed a size of [~]3.4 Mbp and 63% GC content. FWC3 is a coccoid, pale pink pigmented methanotroph and is seen in the form of diplococci, triplets, tetrads or small aggregates. After comparison of the complete 16S rRNA gene sequence, average amino-acid similarities and digital DNA-DNA hybridization values with that of the neighboring type species, we propose that the strain belongs to a novel genus and species, Ca. Methylolobus aquaticus FWC3Ts.

Strain LLG6346-3.1T, isolated from the thallus of the brown alga Ericaria zosteroides collected in Mediterranean Sea near Bastia in Corsica, France, was characterized using a polyphasic method. Cells were Gram-stain-negative, strictly aerobic, non-flagellated, motile by gliding, rod-shaped and grew optimally at 30-33 {degrees}C, at pH 8-8.5 and with 4-5 % NaCl. Strain LLG6346-3.1T used the seaweed polysaccharide alginic acid as sole carbon source which was vigorously liquefied. Phylogenetic analyses showed that the bacterium is affiliated to the genus Zobellia (family Flavobacteriaceae, class Flavobacteriia). Strain LLG6346-3.1T exhibited 16S rRNA gene sequence similarity values of 98.5 and 98.3 % to the type strains of Zobellia russellii and Zobellia roscoffensis respectively, and of 97.4-98.2 % to other species of the genus Zobellia. The DNA G+C content of strain LLG6346-3.1T was determined to be 38.28 mol%. Digital DNA-DNA hybridization predictions by the ANI and GGDC methods between strain LLG6346-3.1T and other members of the genus Zobellia showed values of 76-88 %, and below 37 %, respectively. The phenotypic, phylogenetic and genomic analyses show that strain LLG6346-3.1T is distinct from species of the genus Zobellia with validly published names and that it represents a novel species of the genus Zobellia, for which the name Zobellia alginoliquefaciens sp. nov. is proposed. The type strain is LLG6346-3.1T (RCC 7657T = LLG 32918T).

Besides an industrial pollutant, 2,4-dinitrophenol (DNP) has been used illegally as a weight loss drug that had claimed human lives. Little is known about the metabolism of DNP, particularly among Gram-negative bacteria. In this study, two non-contiguous genetic loci of Paraburkholderia (formerly Burkholderia) sp. strain KU-46 genome were identified and four key initial genes (dnpA, dnpB, and dnpC1C2) were characterized to provide molecular and biochemical evidence for the degradation of DNP via the formation of 4-nitrophenol (NP), a pathway that is unique among DNP utilizing bacteria. Reverse transcription PCR analysis indicated that the dnpA gene encoding the initial hydride transferase (28 kDa), and the dnpB gene encoding a nitrite-eliminating enzyme (33 kDa), are inducible by DNP and the two genes are organized in an operon. Purified DnpA and DnpB from overexpression clones in Escherichia coli effected the transformation of DNP to NP via the formation of hydride-Meisenheimer complex of DNP. The function of DnpB appears new since all homologs of DnpB sequences in the protein database are annotated as putative nitrate ABC transporter substrate-binding proteins. The gene cluster responsible for the degradation of DNP after NP formation was designated dnpC1C2DXFER. DnpC1 and DnpC2 were functionally characterized as the respective FAD reductase and oxygenase components of the two-component NP monooxygenase. Both NP and 4-nitrocatechol were shown to be substrates, producing hydroquinone and hydroxyquinol, respectively. Elucidation of the hqdA1A2BCD gene cluster allows the delineation of the final degradation pathway of hydroquinone to {beta}-ketoadipate prior to its entry to the tricarboxylic acid cycle.\n\nImportanceThis study fills a gap in our knowledge and understanding of the genetic basis and biochemical pathway for the degradation of 2,4-dinitrophenol (DNP) in Gram-negative bacteria, represented by the prototypical Paraburkholderia sp. strain KU-46 that metabolizes DNP through the formation of 4-nitrophenol, a pathway unseen by other DNP utilizers. The newly cloned genes could serve as DNA probes in biomonitoring as well as finding application in new biocatalyst development to access green chemicals. By and large, knowledge of the diverse strategies used by microorganisms to degrade DNP will contribute to the development of bioremediation solutions since DNP is an industrial pollutant used widely in the chemical industry for the synthesis of pesticides, insecticides, sulfur dyes, wood preservatives, and explosives, etc. (119 words)

A facultatively anaerobic, Gram-stain-negative, non-motile, curved rod-shaped bacterium, designated WC007T, was isolated from the deep-sea cold seep, P. R. China. Strain WC007T was found to grow at temperatures from 28 to 37 {degrees}C (optimum, 30 {degrees}C), at pH values between pH 6.0 and 8.0 (optimum, pH 7.0) and in 0-5.0% (w/v) NaCl (optimum, 1.0%). The major fatty acids (>10.0%) were iso-C15:0, C16:0, summed feature 3 and summed feature 8. The major isoprenoid quinone was MK-7. Predominant polar lipids were phosphatidylethanolamine, one unidentified phospholipid, one unidentified aminolipid and one unidentified lipid. The G+C content of the genomic DNA was 38.38%. The average nucleotide identity (ANIb and ANIm), amino acid identity (AAI), the tetranucleotide signatures (Tetra) and in silico DNA-DNA hybridization (isDDH) similarities between the genome sequences of isolate WC007T and Maribellus luteus XSD2T were 70.11%, 84.94%, 71.0%, 0.92022 and 20.40%, respectively, indicating that strain WC007T was distinguished from M. luteus. Phylogenetic analysis based on 16S rRNA gene sequences placed strain WC007T within the genus Maribellus and showed the highest similarity to strain XSD2T (95.70%). In combination of the results of phylogenetic analysis and phenotypic and chemotaxonomic data, strain WC007T was considered to represent a novel species of the genus Maribellus, for which the name Maribellus comscasis sp. nov. is proposed. The type strain is WC007T (=KCTC 25169T = MCCC 1K04777T). The available of the genome sequence of strain WC007T would be helpful in understanding the degradation mechanism of difficult-to-degrade polysaccharides.

Methanotrophic bacterial isolates were identified in this study using the molecular detection method, isolated using microbiological techniques, and studied their cellular shape using atomic force microscopy. Two methanotrophic bacterial species belonging to the Methylocaldum and Methylomonas genera were provisionally designated as Isolate 1 and Isolate 5, thus isolated from the Oil-Natural Gas Field and Paddy Field, respectively. The Oil-Natural Gas Field Isolate 1 showed 91.82-97.25% sequence homology to the reference Methanotrophic species, whereas Paddy Field Isolate 5 showed 79.72-84.99% sequence homology to the reference Methylomonas species in the NCBI database. As per the phylogenetic analysis, Oil-Natural Gas Field Isolate 1 and Paddy Field Isolate 5 are possibly new species of Methylocaldum and Methylomonas genus, respectively. In addition, the microscopic study also supported the molecular identification and phylogenetic analysis of isolated species by showing the cocci and rod shapes for the Oil-Natural Gas Field Isolate 1 and Paddy Field Isolate 5, respectively.

A motile, Gram-stain-positive, rod-shaped, non-sporing, tolerate up to 5% NaCl, grew at 0-25 {degrees}C, designated Exiguobacterium sp. HA2 was isolated from the soil of the Ilam Mountains of Iran during October 2016. The major isoprenoid quinone is MK-7 and in the smaller amount are MK-6 and MK-8. Polar lipids included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine. Major fatty acids (>10 %) are isoC13:0, isoC15:0 and C16:0. The bacterial cell wall peptidoglycan layer was lysine-glycine. The 16S rRNA sequence was analyzed at the phylogenetic levels. Also, A supplemental comparison was made between five other genes including csp, gyrB, hsp70, rpoB, and citC. According to the results of genotypic and phenotypic characteristics, the strain was categorized in the genus Exiguobacterium. This bacterium had the closest relation with Exiguobacterium undae, and thus was dubbed Exiguobacterium sp. HA2. The different in the Phenotypic, functional characteristics and phylogenetic indicated Exiguobacterium sp. HA2 can be regarded as representing considered a novel species within the genus Exiguobacterium.

A novel bacterial strain designated ADMK78T was isolated from the saline desert soil. The cells were rod-shaped, Gram-negative, and non-motile. The strain ADMK78T grows best at 28{degrees}C and pH 7.0 and can tolerate up to 2% (w/v) NaCl. Based on 16S rRNA gene phylogeny, the strain ADMK78T belongs to the genus Rhizobium, with the highest similarity to Rhizobium wuzhouense W44T (98.7%) and Rhizobium ipomoeae shin9-1T (97.9%). Core-genes based phylogenetic analysis revealed that the strain ADMK78T forms a distinct branch in between Rhizobium ipomoeae shin9-1T and Rhizobium selenitireducens BAA-1503T. The average nucleotide identity of ADMK78T was less than 82%, to members of the family Rhizobiaceae. The genomic DNA G+C content of strain ADMK78T is 58.6 mol%. The major fatty acids of strain ADMK78T were C18:0 and C18:1 {omega}7c. The strain ADMK78T showed differences in physiological, phenotypic, and protein profiles estimated by MALDI-TOF MS to its closest relatives. Based on the phenotypic, chemotaxonomic properties, and phylogenetic analyses, the strain ADMK78T could be distinguished from the recognized species of the genus Rhizobium. It is suggested to represent a novel species of this genus, for which the name Rhizobium desertarenae sp. nov. is proposed. The type strain is ADMK78T (=MCC 3400T; KACC 21383T; JCM 33657T).

ASS1 was isolated as a motile Stenotrophomonas strain from crude oil-contaminated soils in Tabasco, Mexico. We characterized this strain using physiological and biochemical traits. ASS1 grew at temperature 25-37 (optimally at 37 {degrees} C) and at pH 6 to 8 (optimally at pH 7 to 8). The assembled genome has a total length of 4.56MB with a G + C content of 66.6%. The 16S rRNA gene sequence analysis confirmed that this strain belongs to the genus Stenotrophomonas. Based on the 16S rRNA analysis, Stenotrophomonas geniculata ATCC 19374 is the closest species, and it shares 99.86% similarity with ASS1. Similarly, a phylogenomic tree based on core genome sequence revealed that the closest species to ASS1 is Stenotrophomonas geniculata ATCC 19374. The major fatty acids in ASS1 are C16:0, antesio C15:0, iso C12:0, iso C15:0, iso C17:0 and C18:0. The genome of ASS1 consists of 4,373,402 bp. The Average Nucleotide Identity (ANI) values for ASS1 which it shared with its closest phylogenetic neighbors, are Stenotrophomonas geniculata ATCC 19374 = JCM 13324 [T] 92.66 %, Stenotrophomonas maltophilia 13637[T] 92.15%, Stenotrophomonas maltophilia K279a 92.13% Stenotrophomonas maltophilia R551-3 92.15% Stenotrophomonas maltophilia MTCC 434 [T] 92.08% and Pseudomonas hibisicicola ATCC [T] 91.66%. ASS1 possesses genes that are essential for the degradation of Polycyclic Aromatic hydrocarbon. Genes such as 1, 2 dihydroxyl 1, 2 dihydronaphthalene dehydrogenase; MG068 17425, homologous to 2 hydroxyl chromene 2 carboxylate isomerases; MG 18055, homologous to salicylaldehyde dehydrogenase and MG068 20095, homologous to naphthalene 1, 2 dioxygenases were identified in ASS1. The dDDH value between ASS1 and its closest neighbor Stenotrophomonas geniculata ATCC 19374 = JCM 13324 [T] is 50%, which is the highest for all the typed species and as such we proposed that ASS1 is a novel species with the name Stenotrophomonas oleivorans sp. nov. sp. nov. and ASS1T as the typed strain

Citrobacter spp. are facultative anaerobic, Gram-negative bacteria that have been known to cause infections in the urinary tract and bloodstream. These infections are predominantly nosocomial in nature. Hospitals are considered to be reservoirs for multiresistant bacteria; as such, their wastewater microbiome is characterized by these bacteria. In the present study, a carbapenem-resistant Citrobacter braakii strain was isolated from hospital sewage in Greifswald, Germany. This particular strain, designated C. braakii GW-Imi-1b1, was the focus of a comprehensive investigation including analyses of antibiotic resistances and proteome profile. Antibiotic resistance tests revealed a broad resistance spectrum, including chloramphenicol, tetracycline, and ertapenem. In addition, an investigation into the defense mechanisms revealed a constitutive presence of a {beta}-lactamase at an unusually high concentration. The growth curve with different meropenem concentrations revealed almost no effect on growth at 20 {micro}g/ml meropenem. Consequently, the high prevalence of {beta}-lactamase may be a contributing factor to the observed carbapenem resistance. Furthermore, due to its localization on the plasmid, it may be capable of being transferred within the wastewater microbiome. Antibiotic treatment of bacterial infections can be substituted or accompanied by phage therapy. The lytic phage vB_CbrP_HGW_001 was isolated from sewage and has been shown to be capable of lysing C. braakii GW-Imi-1b1. Sequencing and microscopic analysis revealed a Kayfunavirus within the Caurovicetes. Subsequent properties of vB_CbrP_HGW_001 indicated the potential for a promising alternative treatment to antibiotics. Phage therapy, particularly when employed in combination with antibiotics, demonstrates a valuable tool in the combat against infections caused by multidrug-resistant bacteria.

Two mesophilic, hydrogenotrophic methanogens, WWM1085 and M. smithii GRAZ-2 were isolated from human fecal samples. WWM1085 was isolated from an individual in the USA, and represents a novel species with in the genus Methanobrevibacter. M. smithii GRAZ-2 (= DSM 116045) was retrieved from fecal samples of a European, healthy female and represents a novel strain within this genus. Both Methanobrevibacter representatives form non-flagellated, short rods with variable morphologies and the capacity to form filaments. Both isolates showed the typical fluorescence of F420 and methane production. Compared to M. smithii GRAZ-2, WWM1085 did not accumulate formate when grown on H2 and CO2. The optimal growth conditions were at 37{degrees}C, and pH 7. Full genome sequencing revealed a genomic difference of WWM1085 to the type strain of M. smithii PS (type strain; DSM 861), with 93.55% ANI and major differences in the sequence of its mcrA gene (3.3% difference in nucleotide sequence). Differences in the 16S rRNA gene were very minor and thus distinction based on this sequence might not be possible. M. smithii GRAZ-2 was identified as a novel strain within the Methanobrevibacter genus (ANI 99.04 % to M. smithii PS). Due to the major differences of WWM1085 and M. smithii type strain PS in phenotypic, genomic and metabolic features, we propose M. intestini sp. nov. as a novel species with WWM1085 as the type strain (DSM 116060T = CECT 30992).

A strictly aerobic, Gram-stain-negative, rod-shaped and motile bacterium, designated strain KMM 296, isolated from the coelomic fluid of mussel Crenomytilus grayanus, was investigated in details due to its ability to produce a highly active alkaline phosphatase of the structural family PhoA. A previous taxonomic study placed the strain to the species Cobetia marina, a member of the family Halomonadaceae of the class Gammaproteobacteria. However, the comprehensive phylogenetic analysis based on 16S rRNA gene sequencing revealed that the strain KMM 296 is most closely related to Cobetia amphilecti NRIC 815T with the 16S rRNA gene sequence similarity of 100%. The mussel isolate grew with 0.5-19% NaCl and at 4 - 42{degrees}C and hydrolysed Tweens 20 and 40, and L-tyrosine. The DNA G+C content was 62.5 mol%. The prevalent fatty acids were C18:1 {omega}7c, C12:0 3-OH, C18:1 {omega}7c, C12:0 and C17:0 cyclo. The polar lipid profile was characterized by the presence of phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, and unidentified aminolipid, phospholipid, and lipids. The major respiratory quinone was Q-8. According to phylogenetic evidence and similarity in the chemotaxonomic and genotypic properties of the mussel isolate and its nearest neighbors, the strain KMM 296 represents a member of the species C. amphilecti. A comparative analysis of the type strains genomes of the species C. amphilecti and C. litoralis showed that they belong to a single species. In addition, a high similarity of the genome sequences of C. pacifica NRIC 813T and C. marina LMG 2217T also allows suggesting the affiliation of these two species to one species. Based on the rules of priority, C. litoralis should be reclassified as a later heterotypic synonym of C. amphilecti, and C. pacifica is a later heterotypic synonym of C. marina. The emended descriptions of the species C. amphilecti and C. marina are also proposed.

A taxonomic re-evaluation of the Antarctic psychrotrophic bacterium Pseudomonas syringae Lz4W was performed in the light of its available genome sequence and due to a revision in the key phenotypic characteristics that are in conflict with the "syringae" group of Pseudomonads. A 16S rRNA gene sequence based phylogenetic analysis suggested that Lz4WT strain belongs to "fragi" cluster of Pseudomonas species, with closest similarity (99.72%) to the type strain P. deceptionensis M1T. However, in silico analysis of the Lz4W genome sequence using SpecI (species identification tools), ANI (average nucleotide identity), and GBDP (Genome Blast Distance phylogeny) methods suggest that Lz4WT strain cannot be delineated with any of the type strains of "fragi" cluster of species. Based on predictive low DNA-DNA hybridization value (<29.9%) and differences in phenotypic features with the related species we suggest that Lz4WT is a novel species under the Pseudomonas genus, and we propose that the strain be named as Pseudomonas cryophila sp. nov. The type strain is Lz4WT (=CFBP 8403T =KCTC 42933T =LMG 29591T =MTCC 673T).

The family Natrialbaceae is a member of the class Halobacteria of the archaeal phylum Euryarchaeota. Seventeen genera with validly or effectively published names are currently included within this family. In this study, using pairwise average nucleotide identity and average amino acid identity comparisons in conjunction with phylogenetic analysis, it has been shown that the family Natrialbaceae is highly diverse and contains several potentially novel species and genera that are yet to be fully characterized. The deduced proteome sequence-based phylogenetic tree, constructed using the alignment- and parameter-free method CVTree3, contained six major clades, with Salinarchaeum sp. Harcht-Bsk1 being the only representative within clade 1. Furthermore, Haloterrigena daqingensis was found to be closely related to Natronorubrum sediminis, and it is proposed that these archaea together represent a novel genus. Interestingly, Haloterrigena jeotgali, Haloterrigena thermotolerans, and Natrinema pellirubrum were found to be very closely related to each other, and it is proposed that they be merged into a single species. Notably, the type genus Natrialba itself appeared to be heterogenous and contains species that could be broadly classified among two genera. Likewise, the genus Natrinema is also heterogenous and contains species that could be classified among six genera. Altogether, 19 novel genera have been proposed to be created, and four haloalkaliphilic archaea hitherto recognized only using genus names are confirmed to represent novel species.

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=85 SRC="FIGDIR/small/439031v1_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@1ed7edaorg.highwire.dtl.DTLVardef@1234542org.highwire.dtl.DTLVardef@97f6cborg.highwire.dtl.DTLVardef@cb8cce_HPS_FORMAT_FIGEXP M_FIG C_FIG The morphology, 16S rRNA gene phylogeny and the 16S-23S rRNA gene ITS secondary structures of three strains of marine Cyanobacteria, isolated from inter- and subtidal environments from north Portugal were studied, resulting in the description of Zarkia subtidalensis gen. et. sp. nov. (Zarkiaceae fam. nov.) and Romeriopsis marina gen. et. sp. nov (Leptolyngbyaceae). No diacritical morphological characters were found either for the new family or for the new genera. The 16S rRNA gene Maximum Likelihood and Bayesian phylogenies supported that Zarkia and Zarkiaceae are members of the Oscillatoriales, positioned close to Microcoleaceae genera, but distant from Microcoleus. Romeriopsis is positioned within the Leptolyngbyaceae and is closely related to Alkalinema. The secondary structures of the D1-D1, Box B, V2 and V3 helices corroborate with the phylogenetic results. Furthermore, our study supports previous observations of polyphyletic Oscillatoriales families and reinforces the need for their taxonomical revision.

The present study described the comparative genomic analysis of the validly named species of the genus Pseudomonas to define the taxonomic assignment. Genomic information for 208 type strains was available in the NCBI genome database at the time of conducting this analysis. The ANI, AAI and in silico DNA DNA hybridization (isDDH) data were higher than the threshold values for the twelve strains with their closely related type species. Whole genome comparisons shared 97 - 99 % average nucleotide identity, 97.85 to 99.19 % average amino acid identity and 72.80 to 90.40 % digital DNA DNA hybridization values. Further, the phylogenomic analysis based on the core genome confirmed that P. humi CCA1 and P. citronellolis LMG 18378, P. zeshuii KACC 15471 and P. luteola NBRC 103146, P. oryzihabitans DSM 6835 and P. psychrotolerans DSM 15758, P. nitroreducens DSM 14399 and P. nitritireducens WZBFD3-5A2, P. fluvialis CCM 8778 and P. pharmacofabricae ZYSR67-Z, P. panacis DSM 18529 and P. marginalis DSM 13124 formed a monophyletic clade. Thus, we proposed six type species viz., P. humi CCA1, P. zeshuii KACC 15471, P. psychrotolerans DSM 15758, P. nitritireducens WZBFD3 5A2, P. pharmacofabricae ZYSR67 Z and P. panacis DSM 18529 are the later heterotypic synonym of P. citronellolis Lang 2007, P. luteola, P. oryzihabitans, P. nitroreducens Lang 2007, P. fluvialis and P. marginalis (Brown 1918) Stevens 1925 (Approved Lists 1980), respectively considering the priority date of publication.

A Gram-negative bacterial strain, designated ACPtT, was isolated from the covering top soil of an active charcoal burning pile. Cells of ACPtT were strictly anaerobic, rod-shaped and grew optimally at 40{degrees}C and pH 7. The substrates ribose, glucose, sucrose, raffinose, melezitose, pyruvate, vanillate, syringate, methanol and CO were utilized for growth. Phylogenomic analysis of the 4.1 Mb genome showed that strain ACPt represented a novel species of the genus Sporomusa. The most closely related species to ACPtT was Sporomusa malonica with an average amino acid identity of 80.1%. The genome of ACPtT encoded cytochromes, ubiquinones, the Wood-Ljungdahl gene cluster and an Rnf complex, which were identified as common features all Sporomusa type strains. However, strain ACPt did not ferment H2 + CO2 via acetogenesis as other Sporomusa species, but employed the metabolism of a carboxydotrophic hydrogenogen converting CO to H2 + CO2. Based on the genomic, morphological and physiological features presented in this study, strain ACPtT is proposed as a novel species in the genus Sporomusa with the name Sporomusa carbonis sp. nov. (DSM 116159T and CCOS 2105T).

In our phylogenetic studies of a bacterial strain isolated from an Arabidopsis suspension culture, we obtained convincing evidence for contamination of the published 16S rRNA gene sequence of Rothia amarae type strain J18T (GenBank AY043359.1). Correction of this sequence by deleting the contamination region eliminated contradictions in bioinformatic results that included comparisons of the small-subunit (SSU) rRNA secondary structures. Further, correction of the contaminated sequence yielded sequence identity values for the 16S rRNA genes of the isolate and R. amarae type strain J18T (and more than a dozen other R. amarae members) above the threshold for species demarcation. The phylogram of the 16S rRNA gene sequences of the type strains closely related to the isolate under study (interspecies sequence identity values, 96-98.7%) united members of the family Micrococcaceae (the genera Rothia, Kocuria and Arthrobacter). A great diversity of habitat conditions was noted for these bacteria, isolated from animals, soil, aqueous media, plant tissues and other sources. This applies, in particular, to R. amarae members that are most closely related to the isolate under study by the 16S rRNA gene sequence criterion (sequence identity, near 100%) and belong to four ecotypes: Antarctic, aquatic, soil and endophytic.