With the progress of biotechnological research and improvements
made in bioprocessing with pure cultures, microbial
consortia have gained recognition for accomplishing biological
processes with improved effectiveness. Microbes are
indispensable tool in developing bioprocesses for the production
of bioenergy and biochemicals while utilizing renewable
resources due to technical, economic and environmental
advantages. They communicate with specific cohorts
in close proximity to promote metabolic cooperation. Use of
positive microbial associations has been recognized widely,
especially in food industries and bioremediation of toxic compounds
and waste materials. Role of microbial associations
in developing sustainable energy sources and substitutes for
conventional fuels is highly promising with many commercial
prospects. Detoxification of chemical contaminants sourced
from domestic, agricultural and industrial wastes has also been
achieved through microbial catalysis in pure and co-culture
systems. Methanotrophs, the sole biological sink of greenhouse
gas methane, catalyze the methane monooxygenasemediated
oxidation of methane to methanol, a high energy
density liquid and key platform chemical to produce commodity
chemical compounds and their derivatives. Constructed
microbial consortia have positive effects, such as improved
biomass, biocatalytic potential, stability etc. In a methanotroph-
heterotroph consortium, non-methanotrophs provide
key nutrient factors and alleviate the toxicity from the culture.
Non-methanotrophic organisms biologically stimulate the
growth and activity of methanotrophs via production of growth
stimulators. However, methanotrophs in association with cocultured
microorganisms are in need of further exploration
and thorough investigation to study their interaction mode
and application with improved effectiveness.
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J. Microbiol. 2019;57(11):953-958. Published online August 28, 2019
A strictly aerobic, motile, endospore-forming, rod-shaped
bacterium, designated HS21T, was isolated from rhizospheric
soil of the Korean fir tree (Abies koreana) from Halla mountain
on Jeju island, Korea. Growth of strain HS21T was observed
at pH 6.0–8.0 (optimum: pH 7.0), 0–2% (w/v) NaCl
and 4–30°C (optimum: 25°C). A comparative analysis of 16S
rRNA gene sequences showed that strain HS21T was most
closely related to Cohnella luojiensis HY-22RT (97.6%), followed
by C. lupini RLAHU4BT (97.4%) and C. collisoli NKM-
5T (97.2%). The genome of strain HS21T comprised a circular
chromosome of 7,059,027 bp with 44.8% G + C content. The
DNA-DNA relatedness values between strain HS21T and C.
luojiensis HY-22RT and C. lupini RLAHU4BT were 18.1% and
13.8%, respectively. The major cellular fatty acids (> 5%) of
the isolate were anteiso-C15:0, iso-C16:0, C16:0, and iso-C15:0. The
polar lipids present were diphosphatidylglycerol, phosphatidylglycerol,
phosphatidylethanolamine, lysylphosphatidylglycerol,
and three unidentified aminophospholipids. Based
on its phenotypic, phylogenetic, genomic, and chemotaxonomic
properties, strain HS21T represents a novel species of
the genus Cohnella, for which the name Cohnella abietis sp.
nov. is proposed. The type strain is HS21T (= KCTC 43028T
= CCTCC AB 2019010T).
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A Gram-stain-negative, asporogenous, aerobic rods, motile by
means of a single polar flagellum, catalase- and oxidase-positive,
methylotrophic bacterium, designated 17Sr1-28T, was
isolated from gamma ray-irradiated soil. The 16S rRNA gene
sequence analysis showed that strain 17Sr1-28T was phylogenetically
related to Methylobacterium currus PR1016AT (96.8%),
Methylobacterium platani PMB02T (96.2%), Methylobacterium
aquaticum DSM 16371T (96.3%), Methylobacterium tarhaniae
N4211T (96.4%), Methylobacterium frigidaeris IER25-16T
(95.8%), and Methylobacterium organophilum JCM 2833T
(92.7%). The G+C content calculated based on genome sequence
was 71.6%. The average nucleotide identity and in
silico DNA-DNA hybridization values between strain 17Sr1-
28T and M. currus, M. platani, M. aquaticum, M. tarhaniae,
M. frigidaeris, and M. organophilum were 77.7–90.4% and
22–39.6%, respectively. The major fatty acids of strain 17Sr1-
28T were summed feature 8 (C18:1 ω7c and/or C18:1 ω6c), and
summed feature 3 (C16:1 ω7c and/or C16:1 ω6c). The predominant
quinone was ubiquinone 10 and the major polar lipids
were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine,
and phosphatidylglycerol. On the basis
of the data from phenotypic tests and genotypic differences
between strain 17Sr1-28T and its close phylogenetic relatives,
strain 17Sr1-28T represents a new species belonging to the
genus Methylobacterium, for which the name Methylobacterium
terrae sp. nov. (= KCTC 52904T = NBRC 112873T) is
proposed.
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A polyphasic taxonomy approach was used to characterize
strain YBJ-36T, isolated from a freshwater lake in Taiwan.
Phylogenetic analyses, based on 16S rRNA gene sequences
and coding sequences of an up-to-date bacterial core gene
set (92 protein clusters), indicated that strain YBJ-36T formed
a phylogenetic lineage in the genus Mucilaginibacter. 16S
rRNA gene sequence similarity indicated that strain YBJ-36T
is closely related to species within the genus Mucilaginibacter
(93.8–97.8% sequence similarity) and is most similar to Mucilaginibacter
fluminis TTM-2T (97.8%), followed by Mucilaginibacter
roseus TTM-1T (97.2%). Microbiological analyses
demonstrated that strain YBJ-36T is Gram-negative, aerobic,
non-motile, rod-shaped, surrounded by a thick capsule, and
forms pink-colored colonies. Strain YBJ-36T grew between
20–40°C (optimal range, 35–37°C), pH 5.5–7.0 (optimal pH
of 6) and 0–2% NaCl (optimal concentration, 0.5%). The predominant
fatty acids of strain YBJ-36T are iso-C15:0 and summed
feature 3 (C16:1 ω7c and/or C16:1 ω6c), the major polar lipid
is phosphatidylethanolamine, the major polyamine is homospermidine,
and the major isoprenoid quinone is MK-7.
The draft genome is approximately 4.63 Mb in size with a
G+C content of 42.8 mol%. Strain YBJ-36T exhibited less than
35% DNA-DNA relatedness with Mucilaginibacter fluminis
TTM-2T and Mucilaginibacter roseus TTM-1T. Based on phenotypic
and genotypic properties and phylogenetic inference,
strain YBJ-36T should be classified in a novel species of the
genus Mucilaginibacter, for which the name Mucilaginibacter
limnophilus sp. nov. is proposed. The type strain is YBJ-36T
(= BCRC 81056T = KCTC 52811T = LMG 30058T).
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A novel, Gram-stain-negative, marine bacterium, designated
GH2-6T, was isolated from a rhizosphere mudflat of a halophyte
(Carex scabrifolia) collected in Gangwha Island, the
Republic of Korea. The cells of the organism were strictly
aerobic, oxidase- and catalase-positive, non-flagellated rods.
Growth occurred at 20–45°C, pH 5–10, and 0.5–9 (w/v) NaCl.
The requirement of Na+ for growth (0.5–3%) was observed.
The major respiratory quinone was Q-10. The major polar
lipids were phosphatidylcholine, phosphatidylethanolamine,
phosphatidylglycerol, an aminolipid and a glycolipid. The
predominant fatty acids were C18:1 ω7c, C18:0, C16:0, C19:0 cyclo
ω8c, C18:1 ω7c 11-methyl and summed feature 2 (C14:0 3-OH
and/or C16:1 iso I). The genome size was 4.45 Mb and the G+C
content of the genomic DNA was 61.9 mol%. Phylogenetic
analyses based on 16S rRNA gene sequences revealed that
strain GH2-6T belonged to genus Martelella and formed a tight
cluster with M. radicis BM5-7T and M. endophytica YC6887T.
Levels of 16S rRNA gene sequence similarity between the novel
isolate and members of the genus were 99.3–95.5%, but strain
GH2-6T possessed an extended loop (49 nucleotides in length)
between positions 187 and 213 of the 16S rRNA gene sequence
(E. coli numbering). DDH values in vitro between the novel
isolate and the closest relatives were 23.2±12.8 – 46.3±5.2%.
On the basis of polyphasic data presented in this study, the
type strain GH2-6T (= KACC 19403T = KCTC 62125T = NBRC
113212T) represents a novel species of the genus Martelella
for which the name Martelella lutilitoris sp. nov. is proposed.
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International Journal of Systematic and Evolutionary Microbiology
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Microalgae and bacteria are known to be closely associated in
diverse environments. To isolate dominant bacterial species
associated with a green alga, Dunaliella tertiolecta, a photoreactor
culture of the microalga was investigated using culture-
based and culture-independent approaches. The bacterial
community structure of the algal culture showed that
the most abundant bacterial species under the culture conditions
was related to the genus Winogradskyella. The closely
related amplicon sequences, showing ≥ 99.5% 16S rRNA gene
sequence similarity to one of the isolates, designated IMCC-
33238T, constituted > 49% of the bacterial community and
was therefore regarded as the most dominant species in the
algal culture. Strain IMCC33238T was characterized by Gramstaining-
negative and orange-colored rods. Phylogenetic analyses
of the 16S rRNA genes as well as whole genome sequences
revealed that strain IMCC33238T belonged to Winogradskyella
and shared more than 97.2% 16S rRNA gene sequence
similarity with Winogradskyella species. The strain
contained iso-C15:1 G, iso-C15:0, iso-C15:0 3-OH, and summed
feature 3 (C16:1 ω6c and/or C16:1 ω7c) as major fatty acids and
MK-6 as the predominant quinone. The polar lipids found
in strain IMCC33238T were phosphatidylethanolamine, two
unidentified aminolipids, and three unidentified lipids. The
genome of strain IMCC33238T was 3.37 Mbp in size with
33.9 mol% G + C content and proteorhodopsin. Many genes
encoding folate and vitamin production are considered to play
an important role in the bacteria-algae interaction. On the
basis of phylogenetic and phenotypic characteristics, strain
IMCC33238T represents a novel species in the genus Winogradskyella,
for which the name Winogradskyella algicola sp.
nov. is proposed. The type strain is IMCC33238T (= KACC
21192T = NBRC 113704T).
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Taxonomic study of nine new Winogradskyella species occurring in the shallow waters of Helgoland Roads, North Sea. Proposal of Winogradskyella schleiferi sp. nov., Winogradskyella costae sp. nov., Winogradskyella helgolandensis sp. nov., Winogradskyella v Carlota Alejandre-Colomo, Tomeu Viver, Mercedes Urdiain, Ben Francis, Jens Harder, Peter Kämpfer, Rudolf Amann, Ramon Rosselló-Móra Systematic and Applied Microbiology.2020; 43(6): 126128. CrossRef
A Gram-stain-positive, aerobic, motile, and rod-shaped bacterial
strain designated TGS2-1T was isolated from sediment
soil in the Nakdong River, Republic of Korea. The optimal
growth of strain TGS2-1T was observed at 28°C and pH 7.0
without NaCl supplementation. Strain TGS2-1T revealed antibiosis
against various bacteria, including Staphylococcus aureus
KCCM 4051, CCARM 3089 (methicillin resistant strains),
Enterococcus faecalis KCCM 11814, Escherichia coli KCTC
2443, Candida albicans KACC 7270, and Filobasidium neoformans
KCTC 7902. Phylogenetic analyses based on the
16S rRNA gene sequences indicated that strain TGS2-1T belonged
to the genus Brevibacillus and shared 93.8–99.7% sequence
similarity with Brevibacillus species. Whole-genome
sequencing of strain TGS2-1T revealed a genome size of 6.2
Mbp and DNA G + C content of 47.0 mol%. The TGS2-1T
genome shared an average nucleotide identity and digital
DNA-DNA hybridization of 74.6–93.3% and 18.6–67.1%,
respectively, with six related Brevibacillus genomes. The major
fatty acid constituents of strain TGS2-1T were anteiso-C15:0
(62.3%) and anteiso-C17:0 (10.8%). Cells of strain TGS2-1T contained
diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine,
seven unidentified aminophospholipids,
and five unidentified lipids. The isoprenoid quinone
detected in the strain was menaquinone-7 (MK-7). Based on
data obtained from this polyphasic taxonomic study, strain
TGS2-1T represents a novel species belonging to genus Brevibacillus,
for which the name B. antibioticus sp. nov. is proposed.
The type strain is TGS2-1T (= KCCM 90326T = NBRC
113840T = FBCC-B2501).
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A Gram-stain-positive, rod-shaped, alkalitolerant, and halophilic
bacterium–designated as strain NKC3-5T–was isolated
from kimchi that was collected from the Geumsan area
in the Republic of Korea. Cells of isolated strain NKC3-5T
were 0.5–0.7μm wide and 1.4–2.8 μm long. The strain
NKC3-5T could grow at up to 20.0% (w/v) NaCl (optimum
10%), pH 6.5–10.0 (optimum pH 9.0), and 25–40°C (optimum
35°C). The cells were able to reduce nitrate under aerobic
conditions, which is the first report in the genus Salicibibacter.
The genome size and genomic G + C content of
strain NKC3-5T were 3,754,174 bp and 45.9 mol%, respectively;
it contained 3,630 coding sequences, 16S rRNA genes
(six 16S, five 5S, and five 23S), and 59 tRNA genes. Phylogenetic
analysis based on 16S rRNA showed that strain NKC-
3-5T clustered with bacterium Salicibibacter kimchii NKC1-1T,
with a similarity of 96.2–97.6%, but formed a distinct branch
with other published species of the family Bacillaceae. In addition,
OrthoANI value between strain NKC3-5T and Salicibibacter
kimchii NKC1-1T was far lower than the species demarcation
threshold. Using functional genome annotation,
the result found that carbohydrate, amino acid, and vitamin
metabolism related genes were highly distributed in the genome
of strain NKC3-5T. Comparative genomic analysis revealed
that strain NKC3-5T had 716 pan-genome orthologous
groups (POGs), dominated with carbohydrate metabolism.
Phylogenomic analysis based on the concatenated core
POGs revealed that strain NKC3-5T was closely related to
Salicibibacter kimchii. The predominant polar lipids were
phosphatidylglycerol and two unidentified lipids. Anteiso-
C15:0, iso-C17:0, anteiso-C17:0, and iso-C15:0 were the major cellular
fatty acids, and menaquinone-7 was the major isoprenoid
quinone present in strain NKC3-5T. Cell wall peptidoglycan
analysis of strain NKC3-5T showed that meso-diaminopimelic
acid was the diagnostic diamino acid. The phenotypic,
genomic, phylogenetic, and chemotaxonomic properties
reveal that the strain represents a novel species of
the genus Salicibibacter, for which the name Salicibibacter
halophilus sp. nov. is proposed, with the type strain NKC3-5T
(= KACC 21230T = JCM 33437T).
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While several methods for determining postmortem submersion
interval (PMSI) in drowning cases have been suggested,
the estimation of PMSI remains difficult. Next-generation
sequencing (NGS) technology enables simultaneous
identification of multiple taxa from environmental samples.
Although NGS has been applied to estimate time since death,
this application has been mainly focused on terrestrial cases.
As a case study, we investigated microeukaryotic biodiversity
and community structures in submerged car bonnet and
drowned pig using NGS technology. NGS analysis showed
that the microeukaryotic biodiversity in pig carcass was relevantly
lower than that in car bonnet. NGS results also revealed
that water molds and algae were related to decomposition.
Relative abundances of Filobasidium, Achlya, Saprolegnia,
Hydrodicton, Lobosphaera, and Scenedesmus varied
with decomposition period. This data indicated that these
taxa might be useful as good indicators to estimate PMSI.
This study showed microeukaryotic community analysis using
NGS technology may help solve drowning cases in forensic
investigation.
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Although phosphatase and tensin homolog (PTEN) is typically
considered a tumor-suppressor gene, it was recently
suggested that PTEN regulates TLR5-induced immune and
inflammatory responses in intestinal epithelial cells (IECs),
suggesting an immunomodulatory function of PTEN in the
gut. However, this alternative function of PTEN has not yet
been evaluated in an in vivo context of protection against
enteropathogenic bacteria. To address this, we utilized IECrestricted
Pten knockout (PtenΔIEC/ΔIEC) and littermate Pten+/+
mice. These mice were subjected to the streptomycin-pretreated
mouse model of Salmonella infection, and subsequently
given an oral gavage of a low inoculum (2 × 104 CFU)
of Salmonella enterica serovar Typhimurium (S. Typhimurium).
This bacterial infection not only increased the mortality
of PtenΔIEC/ΔIEC mice compared to Pten+/+ mice, but
also induced deleterious gastrointestinal inflammation in
PtenΔIEC/ΔIEC mice manifested by massive histological damage
to the intestinal mucosa. S. Typhimurium infection upregulated
pro-inflammatory cytokine production in the intestine
of PtenΔIEC/ΔIEC mice compared to controls. Furthermore,
bacterial loads were greatly increased in the liver,
mesenteric lymph node, and spleen of PtenΔIEC/ΔIEC mice
compared to controls. Together, these results suggest that
IEC-restricted Pten deficiency renders the host greatly susceptible
to Salmonella infection and support an immuneregulatory
role of PTEN in the gut.
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Enterococci are Gram-positive facultative anaerobic bacteria
that colonize the oral cavity and gastrointestinal tract. Enterococcal
infections, mainly caused by Enterococcus faecalis
and Enterococcus faecium, include apical periodontitis, endocarditis,
and bloodstream infections. Recently, vancomycinresistant
Enterococci are considered major pathogens that
are common but difficult to treat, especially in nosocomial
settings. Moreover, E. faecalis is closely associated with recurrent
endodontic infections and failed endodontic treatment.
In this study, we investigated the effects of short-chain
fatty acids (SCFAs), acetate, propionate, and butyrate, which
are metabolites fermented by gut microbiota, on the growth
of Enterococci. Enterococci were cultured in the presence
or absence of acetate, propionate, or butyrate, and the optical
density at 600 nm was measured to determine bacterial
growth. The minimum inhibitory concentration/minimum
bactericidal concentration test was conducted. Bacteria were
treated with a SCFA, together with clinically used endodontic
treatment methods such as triple antibiotics (metronidazole,
minocycline, and ciprofloxacin) and chlorhexidine gluconate
(CHX) to determine the effects of combination treatment.
Of the SCFAs, propionate had a bacteriostatic effect, inhibiting
the growth of E. faecalis in a dose-dependent manner and
also that of clinical strains of E. faecalis isolated from dental
plaques. Meanwhile, acetate and butyrate had minimal effects
on E. faecalis growth. Moreover, propionate inhibited the
growth of other Enterococci including E. faecium. In addition,
combination treatment of propionate and triple antibiotics
led to further growth inhibition, whereas no cooperative
effect was observed at propionate plus CHX. These results
indicate that propionate attenuates the growth of Enterococci,
suggesting propionate as a potential agent to control
Enterococcal infections, especially when combined with triple
antibiotics.
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ZhenHao Li , Yufang Gong , ShuZe Chen , SiQi Li , Yu Zhang , HuiMin Zhong , ZhouCheng Wang , YiFan Chen , QiXin Deng , YuTing Jiang , LiYing Li , Min Fu , GuoGuo Yi
J. Microbiol. 2019;57(11):1025-1032. Published online August 28, 2019
To compare the ocular surface (OS) microbial communities
and diversity between dry eye (DE) and non-DE (NDE). Furthermore,
we compared meibomian gland dysfunction (MGD)
and non-MGD (NMGD) among DE subjects. The V3-V4 region
of 16S rRNA gene high-throughput sequencing was performed
in the conjunctival swab samples to investigate the
composition of the OS bacterial community in DE (n=35) and
NDE (n=54) and compared the composition of MGD (n=25)
and NMGD (n=10) among DE subjects. Deep sequencing
of OS 16S rDNA from DE (n=35) and NDE (n=54) demonstrated
great a difference in alpha and beta diversity between
the OS bacterial flora (P < 0.05). The similar OS microbial
structures were shown at the phylum and genus levels by bioinformatics
analysis between them, and in LEfSe (linear discriminant
analysis effect size) analysis, Bacteroidia and Bacteroidetes
were enriched in DE, while Pseudomonas was plentiful
in NDE (linear discriminant analysis [LDA] > 4.0). Among
the DE group, there was no significant difference in α and β
diversity between MGD and NMGD (P > 0.05). Surprisingly,
Bacilli was the dominant microbe in MGD, and Bacteroidetes
was the superior bacteria in NMGD among DE subjects (LDA
> 4.0). Different diversity of OS bacteria composition between
DE and NDE and the altered diversity of OS bacteria may
play an important role in DE. Moreover, the lower dominance
of OS bacteria in DE may be associated with the occurrence
and development of DE. Although there was no significant
difference in alpha and beta analysis, the OS dominant microbe
between MGD and NMGD among DE was different.
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Primary infections with the varicella-zoster virus (VZV) result
in varicella, while latent reactivation leads to herpes zoster.
Both varicella and zoster can be prevented by live attenuated
vaccines. There have been reports suggesting that both clinical
VZV strains and those in vaccine preparations are genetically
polymorphic, containing mixtures of both wild-type
and vaccine-type sequences at certain vaccine-specific sites.
In this study, the genetic polymorphism of the VZV genome
was examined by analyzing the frequencies of minor alleles
at each nucleotide position. Next-generation sequencing of
the clinical VZV strain YC02 passaged in an in vitro cell culture
was used to identify genetically polymorphic sites (GPS),
where the minor allele frequency (MAF) exceeded 5%. The
number of GPS increased by 7.3-fold at high passages (p100)
when compared to low passages (p17), although the average
MAF remained similar. GPS were found in 6 open reading
frames (ORFs) in p17, 35, and 54 ORFs in p60 and p100, respectively.
GPS were found more frequently in the dispensable
gene group than the essential gene group, but the average MAF
was greater in the essential gene group. The most common
two major/minor base pairs were A/g and T/c. GPS were found
in all three passages at 16 positions, all located in the reiterated
(R) region. The population diversity as measured by Shannon
entropy increased in p60 and p100. However, the entropy
remained unchanged in the R regions.
Citations
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