Bacillus siralis sp. nov., a novel species from silage with a higher order structural attribute in the 16S rRNA genes.

In 1983, Mycoplasma sp. strain 1220 was isolated in Hungary from the phallus lymph of a gander with phallus inflammation. Between 1983 and 2017, Mycoplasma sp. 1220 was also identified and isolated from the respiratory tract, liver, ovary, testis, peritoneum and cloaca of diseased geese in several countries. Seventeen studied strains produced acid from glucose and fructose but did not hydrolyse arginine or urea, and all grew under aerobic, microaerophilic and anaerobic conditions at 35 to 37 ˚C in either SP4 or pleuropneumonia-like organism medium supplemented with glucose and serum. Colonies on agar showed a typical fried-egg appearance and transmission electron microscopy revealed a typical mycoplasma cellular morphology. Molecular characterization included analysis of the following genetic loci: 16S rRNA, 23S rRNA, 16S–23S rRNA ITS, rpoB, rpoC, rpoD, uvrA, parC, topA, dnaE, fusA and pyk. The genome was sequenced for type strain 1220T. The 16S rRNA gene sequences of studied strains of Mycoplasma sp. 1220 shared 99.02–99.19 % nucleotide similarity with M. anatis strains but demonstrated ≤95.00–96.70 % nucleotide similarity to the 16S rRNA genes of other species of the genus Mycoplasma . Phylogenetic, average nucleotide and amino acid identity analyses revealed that the novel species was most closely related to Mycoplasma anatis . Based on the genetic data, we propose a novel species of the genus Mycoplasma , for which the name Mycoplasma anserisalpingitidis sp. nov. is proposed with the type strain 1220T (=ATCC BAA-2147T=NCTC 13513T=DSM 23982T). The G+C content is 26.70 mol%, genome size is 959110 bp.

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Halosimplex halophilum sp. nov. and Halosimplex salinum sp. nov., isolated from saline soil and a salt mine

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijsem.0.004775 ◽

2021 ◽

Vol 71 (4) ◽

Author(s):

Xiao-Yan Yang ◽

Xue-Meng Yin ◽

Jing Hou ◽

Lin Zhu ◽

Heng-Lin Cui

Keyword(s):

16S Rrna ◽

In Silico ◽

Saline Soil ◽

Novel Species ◽

16S Rrna Genes ◽

Rrna Genes ◽

Threshold Values ◽

Content Type ◽

Link Type ◽

Sequence Similarities

A polyphasic study was undertaken to determine the taxonomic position of two halophilic archaeal strains, TH32T and YPL4T, isolated from saline soil and a salt mine in PR China, respectively. Strains TH32T and YPL4T both have two dissimilar 16S rRNA genes. The two strains exhibited sequence similarities of 91.5–95.5 % for 16S rRNA genes and 90.9 % for the rpoB′ gene. Sequence similarities of 16S rRNA genes and the rpoB′ gene between the two strains and the current four members of Halosimplex were 90.6–97.4 % and 91.4–93.5 %, respectively. Phylogenetic analysis revealed that the two strains formed different branches separating them from the current Halosimplex members. Several phenotypic characteristics differentiate strains TH32T and YPL4T from current Halosimplex members. The polar lipids of the two strains are phosphatidic acid, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and four glycolipids. Two of the glycolipids are chromatographically identical to disulfated mannosyl glucosyl diether and sulfated mannosyl glucosyl diether, respectively, and the remaining two glycolipids are unidentified. The average nucleotide identity (ANI) and in silico DNA–DNA hybridization (DDH) values between the two strains and the current members of Halosimplex (ANI 80.4–89.2 % and in silico DDH 24.0–41.8 %) were much lower than the threshold values proposed as a species boundary, suggesting that the two strains represent novel species of Halosimplex . The values between the two strains (ANI 81.3 % and in silico DDH 24.9 %) were also much lower than the recommended threshold values, which revealed that the two strains represent two genomically different species of Halosimplex . These results showed that strains TH32T (=CGMCC 1.15190T=JCM 30840T) and YPL4T (=CGMCC 1.15329T=JCM 31108T) represent two novel species of Halosimplex , for which the names Halosimplex halophilum sp. nov. and Halosimplex salinum sp. nov. are proposed.

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Streptococcus orisuis sp. nov., isolated from the pig oral cavity

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijs.0.64741-0 ◽

2007 ◽

Vol 57 (6) ◽

pp. 1272-1275 ◽

Cited By ~ 33

Author(s):

Kazuko Takada ◽

Masatomo Hirasawa

Keyword(s):

16S Rrna ◽

Dna Hybridization ◽

Type Strain ◽

Novel Species ◽

Mutans Streptococci ◽

16S Rrna Genes ◽

Rrna Genes ◽

Oral Streptococci ◽

The Novel ◽

Bacterial Strains

Five bacterial strains, designated as NUM 1001T, NUM 1002, NUM 1003, NUM 1004 and NUM 1005, were isolated from the oral cavities of pigs. Colonies grown on mitis salivarius agar were similar in morphology to those of mutans streptococci. The novel isolates were analysed biochemically using the Rapid ID 32 Strep microsystem, subjected to DNA–DNA hybridization with oral streptococci and had their 16S rRNA genes sequenced. On the basis of the phylogenetic and phenotypic evidence obtained, the strains represent a novel species of the genus Streptococcus, for which the name Streptococcus orisuis sp. nov. is proposed. The type strain is NUM 1001T (=JCM 14035T=DSM 18307T).

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Weissella jogaejeotgali sp. nov., isolated from jogae jeotgal, a traditional Korean fermented seafood

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijsem.0.000631 ◽

2015 ◽

Vol 65 (Pt_12) ◽

pp. 4674-4681 ◽

Cited By ~ 18

Author(s):

Se-Hui Lee ◽

Hye-Jin Ku ◽

Min-Ju Ahn ◽

Ji-Sang Hong ◽

Se Hee Lee ◽

...

Keyword(s):

16S Rrna ◽

16S Rrna Gene ◽

Novel Species ◽

Restriction Analysis ◽

16S Rrna Genes ◽

Rrna Genes ◽

Rrna Gene ◽

Carbohydrate Fermentation ◽

Genomic Dnas ◽

Close Relatives

Strain FOL01T was isolated from traditionally fermented Korean jogae jeotgal (fermented clams). Phylogenetic sequence analysis of the 16S rRNA gene from FOL01T revealed that it is closely related to Weissella thailandensis FS61-1T and Weissella paramesenteroides ATCC 33313T with 99.39 % and 98.50 % 16S rRNA gene sequence similarities, respectively. API and VITEK analyses showed that strain FOL01T could be separated from its nearest phylogenetic relatives with respect to carbohydrate fermentation and antibiotic resistance. Subsequent amplified rRNA gene restriction analysis of 16S rRNA genes and HaeIII-restriction enzyme profiling of genomic DNAs revealed different band patterns. In addition, DNA–DNA hybridization of genomic DNAs showed 63.9 % relatedness. Analysis of the composition of cellular fatty acids confirmed that strain FOL01T differs from its close relatives and supports the proposal to assign this organism to a novel species of the genus Weissella. Based on these results, strain FOL01T could be classified as a novel species of the genus Weissella, for which the name Weissella jogaejeotgali sp. nov. is proposed. The type strain is FOL01T ( = KCCM 43128T = JCM 30589T).

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Bartonella jaculi sp. nov., Bartonella callosciuri sp. nov., Bartonella pachyuromydis sp. nov. and Bartonella acomydis sp. nov., isolated from wild Rodentia

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijs.0.041939-0 ◽

2013 ◽

Vol 63 (Pt_5) ◽

pp. 1734-1740 ◽

Cited By ~ 28

Author(s):

Shingo Sato ◽

Hidenori Kabeya ◽

Yuta Fujinaga ◽

Kai Inoue ◽

Yumi Une ◽

...

Keyword(s):

16S Rrna ◽

Phylogenetic Trees ◽

Novel Species ◽

Amino Acid Sequences ◽

16S Rrna Genes ◽

Rrna Genes ◽

Phenotypic Characterization ◽

Sequence Comparisons ◽

Content Type ◽

Link Type

Four novel strains of members of the genus Bartonella , OY2-1T, BR11-1T, FN15-2T and KS2-1T, were isolated from the blood of wild-captured greater Egyptian jerboa (Jaculus orientalis), plantain squirrel (Callosciurus notatus), fat-tailed gerbil (Pachyuromys duprasi) and golden spiny mouse (Acomys russatus). All the animals were imported to Japan as pets from Egypt, Thailand and the Netherlands. The phenotypic characterization (growth conditions, incubation periods, biochemical properties and cell morphologies), DNA G+C contents (37.4 mol% for strain OY2-1T, 35.5 mol% for strain BR11-1T, 35.7 mol% for strain FN15-2T and 37.2 mol% for strain KS2-1T), and sequence analyses of the 16S rRNA genes indicated that those strains belong to the genus Bartonella . Sequence comparisons of gltA and rpoB genes suggested that all of the strains should be classified as novel species of the genus Bartonella . In phylogenetic trees based on the concatenated sequences of five loci, including the 16S rRNA, ftsZ, gltA and rpoB genes and the ITS region, and on the concatenated deduced amino acid sequences of three housekeeping genes (ftsZ, gltA and rpoB), all strains formed distinct clades and had unique mammalian hosts that could be discriminated from other known species of the genus Bartonella . These data strongly support the hypothesis that strains OY2-1T, BR11-1T, FN15-2T and KS2-1T should be classified as representing novel species of the genus Bartonella . The names Bartonella jaculi sp. nov., Bartonella callosciuri sp. nov., Bartonella pachyuromydis sp. nov. and Bartonella acomydis sp. nov. are proposed for these novel species. Type strains of Bartonella jaculi sp. nov., Bartonella callosciuri sp. nov., Bartonella pachyuromydis sp. nov. and Bartonella acomydis sp. nov. are OY2-1T ( = JCM 17712T = KCTC 23655T), BR11-1T ( = JCM 17709T = KCTC 23909T), FN15-2T ( = JCM 17714T = KCTC 23657T) and KS2-1T ( = JCM 17706T = KCTC 23907T), respectively.

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Bartonella japonica sp. nov. and Bartonella silvatica sp. nov., isolated from Apodemus mice

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijs.0.011528-0 ◽

2010 ◽

Vol 60 (4) ◽

pp. 759-763 ◽

Cited By ~ 34

Author(s):

Kai Inoue ◽

Hidenori Kabeya ◽

Hatsumi Shiratori ◽

Kenji Ueda ◽

Michael Y. Kosoy ◽

...

Keyword(s):

16S Rrna ◽

Novel Species ◽

Biochemical Properties ◽

16S Rrna Genes ◽

Rrna Genes ◽

Phenotypic Characterization ◽

Sequencing Analysis ◽

Field Mouse ◽

Bacterial Strains ◽

Apodemus Speciosus

Two bacterial strains, Fuji 18-1T and Fuji 23-1T, were isolated from the blood of the small Japanese field mouse (Apodemus argenteus) and the large Japanese field mouse (Apodemus speciosus), respectively, specimens of which were captured in the forest of Mount Fuji, Japan. Phenotypic characterization (growth conditions, incubation periods, biochemical properties and cell morphologies), DNA G+C contents (40.1 mol% for strain Fuji 18-1T and 40.4 mol% for strain Fuji 23-1T) and sequence analyses of the 16S rRNA genes indicated that both strains were members of the genus Bartonella. Using rpoB and gltA sequencing analysis, the highest sequence similarities between strains Fuji 18-1T, Fuji 23-1T and other recognized species of the genus Bartonella showed values considerably lower than 91.4 % and 89.9 % in the rpoB gene and 89.1 % and 90.4 % in the gltA gene, respectively. It is known that similarities of 95.4 % for the rpoB gene and 96.0 % for the gltA gene can be applied as cut-off values for the designation of novel species of the genus Bartonella. In a phylogenetic tree based on the merged set of concatenated sequences of seven loci [16S rRNA, ftsZ, gltA, groEL, ribC and rpoB genes and the intergenic spacer region (ITS)], strains Fuji 18-1T and Fuji 23-1T formed a distinct clade from other recognized species of the genus Bartonella. These data support the classification of strains Fuji 18-1T and Fuji 23-1T as novel species of the genus Bartonella. The names Bartonella japonica sp. nov. and Bartonella silvatica sp. nov. are proposed for these novel species. The type strains of Bartonella japonica sp. nov. and Bartonella silvatica sp. nov. are Fuji 18-1T (=JCM 15567T=CIP 109861T) and Fuji 23-1T (=JCM 15566T=CIP 109862T), respectively.

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Differentiation Of Clarias Batrachus, C. Gariepinus And Heteropneustes Fossilis By Pcr-Sequencing Of Mitochondrial 16s Rrna Gene

Journal of the Asiatic Society of Bangladesh Science ◽

10.3329/jasbs.v41i1.46190 ◽

2015 ◽

Vol 41 (1) ◽

pp. 51-58

Author(s):

Mohammad Shamimul Alam ◽

Hawa Jahan ◽

Rowshan Ara Begum ◽

Reza M Shahjahan

Keyword(s):

16S Rrna ◽

16S Rrna Gene ◽

Fish Larva ◽

16S Rrna Genes ◽

Rrna Genes ◽

Rrna Gene ◽

Valuable Insight ◽

Multiple Sequence ◽

Pcr Rflp ◽

Alternative Means

Heteropneustesfossilis, Clariasbatrachus and C. gariepinus are three major catfishes ofecological and economic importance. Identification of these fish species becomes aproblem when the usual external morphological features of the fish are lost or removed,such as in canned fish. Also, newly hatched fish larva is often difficult to identify. PCRsequencingprovides accurate alternative means of identification of individuals at specieslevel. So, 16S rRNA genes of three locally collected catfishes were sequenced after PCRamplification and compared with the same gene sequences available from othergeographical regions. Multiple sequence alignment of the 16S rRNA gene fragments ofthe catfish species has revealed polymorphic sites which can be used to differentiate thesethree species from one another and will provide valuable insight in choosing appropriaterestriction enzymes for PCR-RFLP based identification in future. Asiat. Soc. Bangladesh, Sci. 41(1): 51-58, June 2015

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Effects of Supplement of Marichromatium gracile YL28 on Water Quality and Microbial Structures in Shrimp Mariculture Ecosystems

Genes ◽

10.3390/genes12010040 ◽

2020 ◽

Vol 12 (1) ◽

pp. 40

Author(s):

Liang Cui ◽

Bitong Zhu ◽

Xiaobo Zhang ◽

Zhuhua Chan ◽

Chungui Zhao ◽

...

Keyword(s):

Water Quality ◽

16S Rrna ◽

Relative Abundance ◽

Animal Health ◽

Denitrifying Bacteria ◽

Nitrite Reduction ◽

16S Rrna Genes ◽

Rrna Genes ◽

Sequencing Analysis ◽

Shrimp Culture

The elevated NH3-N and NO2-N pollution problems in mariculture have raised concerns because they pose threats to animal health and coastal and offshore environments. Supplement of Marichromatium gracile YL28 (YL28) into polluted shrimp rearing water and sediment significantly decreased ammonia and nitrite concentrations, showing that YL28 functioned as a novel safe marine probiotic in the shrimp culture industry. The diversity of aquatic bacteria in the shrimp mariculture ecosystems was studied by sequencing the V4 region of 16S rRNA genes, with respect to additions of YL28 at the low and high concentrations. It was revealed by 16S rRNA sequencing analysis that Proteobacteria, Planctomycete and Bacteroidetes dominated the community (>80% of operational taxonomic units (OTUs)). Up to 41.6% of the predominant bacterial members were placed in the classes Gammaproteobacteria (14%), Deltaproteobacteria (14%), Planctomycetacia (8%) and Alphaproteobacteria (5.6%) while 40% of OTUs belonged to unclassified ones or others, indicating that the considerable bacterial populations were novel in our shrimp mariculture. Bacterial communities were similar between YL28 supplements and control groups (without addition of YL28) revealed by the β-diversity using PCoA, demonstrating that the additions of YL28 did not disturb the microbiota in shrimp mariculture ecosystems. Instead, the addition of YL28 increased the relative abundance of ammonia-oxidizing and denitrifying bacteria. The quantitative PCR analysis further showed that key genes including nifH and amoA involved in nitrification and nitrate or nitrite reduction significantly increased with YL28 supplementation (p < 0.05). The supplement of YL28 decreased the relative abundance of potential pathogen Vibrio. Together, our studies showed that supplement of YL28 improved the water quality by increasing the relative abundance of ammonia-oxidizing and denitrifying bacteria while the microbial community structure persisted in shrimp mariculture ecosystems.

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Ultra-accurate microbial amplicon sequencing with synthetic long reads

Microbiome ◽

10.1186/s40168-021-01072-3 ◽

2021 ◽

Vol 9 (1) ◽

Author(s):

Benjamin J. Callahan ◽

Dmitry Grinevich ◽

Siddhartha Thakur ◽

Michael A. Balamotis ◽

Tuval Ben Yehezkel

Keyword(s):

Microbial Community ◽

16S Rrna ◽

Amplicon Sequencing ◽

Species Level ◽

Full Length ◽

16S Rrna Genes ◽

Rrna Genes ◽

Strain Identification ◽

Long Reads ◽

Long Read

Abstract Background Out of the many pathogenic bacterial species that are known, only a fraction are readily identifiable directly from a complex microbial community using standard next generation DNA sequencing. Long-read sequencing offers the potential to identify a wider range of species and to differentiate between strains within a species, but attaining sufficient accuracy in complex metagenomes remains a challenge. Methods Here, we describe and analytically validate LoopSeq, a commercially available synthetic long-read (SLR) sequencing technology that generates highly accurate long reads from standard short reads. Results LoopSeq reads are sufficiently long and accurate to identify microbial genes and species directly from complex samples. LoopSeq perfectly recovered the full diversity of 16S rRNA genes from known strains in a synthetic microbial community. Full-length LoopSeq reads had a per-base error rate of 0.005%, which exceeds the accuracy reported for other long-read sequencing technologies. 18S-ITS and genomic sequencing of fungal and bacterial isolates confirmed that LoopSeq sequencing maintains that accuracy for reads up to 6 kb in length. LoopSeq full-length 16S rRNA reads could accurately classify organisms down to the species level in rinsate from retail meat samples, and could differentiate strains within species identified by the CDC as potential foodborne pathogens. Conclusions The order-of-magnitude improvement in length and accuracy over standard Illumina amplicon sequencing achieved with LoopSeq enables accurate species-level and strain identification from complex- to low-biomass microbiome samples. The ability to generate accurate and long microbiome sequencing reads using standard short read sequencers will accelerate the building of quality microbial sequence databases and removes a significant hurdle on the path to precision microbial genomics.

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Microdiversity and phylogeographic diversification of bacterioplankton in pelagic freshwater systems revealed through long-read amplicon sequencing

Microbiome ◽

10.1186/s40168-020-00974-y ◽

2021 ◽

Vol 9 (1) ◽

Author(s):

Yusuke Okazaki ◽

Shohei Fujinaga ◽

Michaela M. Salcher ◽

Cristiana Callieri ◽

Atsushi Tanaka ◽

...

Keyword(s):

16S Rrna ◽

Regional Scale ◽

Scale Up ◽

Amplicon Sequencing ◽

Freshwater Ecosystems ◽

16S Rrna Genes ◽

Rrna Genes ◽

Rrna Gene ◽

Metagenomic Sequencing ◽

Long Read

Abstract Background Freshwater ecosystems are inhabited by members of cosmopolitan bacterioplankton lineages despite the disconnected nature of these habitats. The lineages are delineated based on > 97% 16S rRNA gene sequence similarity, but their intra-lineage microdiversity and phylogeography, which are key to understanding the eco-evolutional processes behind their ubiquity, remain unresolved. Here, we applied long-read amplicon sequencing targeting nearly full-length 16S rRNA genes and the adjacent ribosomal internal transcribed spacer sequences to reveal the intra-lineage diversities of pelagic bacterioplankton assemblages in 11 deep freshwater lakes in Japan and Europe. Results Our single nucleotide-resolved analysis, which was validated using shotgun metagenomic sequencing, uncovered 7–101 amplicon sequence variants for each of the 11 predominant bacterial lineages and demonstrated sympatric, allopatric, and temporal microdiversities that could not be resolved through conventional approaches. Clusters of samples with similar intra-lineage population compositions were identified, which consistently supported genetic isolation between Japan and Europe. At a regional scale (up to hundreds of kilometers), dispersal between lakes was unlikely to be a limiting factor, and environmental factors or genetic drift were potential determinants of population composition. The extent of microdiversification varied among lineages, suggesting that highly diversified lineages (e.g., Iluma-A2 and acI-A1) achieve their ubiquity by containing a consortium of genotypes specific to each habitat, while less diversified lineages (e.g., CL500-11) may be ubiquitous due to a small number of widespread genotypes. The lowest extent of intra-lineage diversification was observed among the dominant hypolimnion-specific lineage (CL500-11), suggesting that their dispersal among lakes is not limited despite the hypolimnion being a more isolated habitat than the epilimnion. Conclusions Our novel approach complemented the limited resolution of short-read amplicon sequencing and limited sensitivity of the metagenome assembly-based approach, and highlighted the complex ecological processes underlying the ubiquity of freshwater bacterioplankton lineages. To fully exploit the performance of the method, its relatively low read throughput is the major bottleneck to be overcome in the future.

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