Alcaligenes faecalis is one of the most important and clinically significant environmental pathogens, increasing in importance due to its isolation from soil and nosocomial environments. The Gram-negative soil bacterium is associated with skin endocarditis, bacteremia, dysentery, meningitis, endophthalmitis, urinary tract infections, and pneumonia in patients. With emerging antibiotic resistance in A. faecalis, it has become crucial to understand the origin of such resistance genes within this clinically significant environmental and gut bacterium. In this research, we studied the impact of antibiotic overuse in poultry and its effect on developing resistance in A. faecalis. We sampled soil and faecal materials from five poultry farms, performed whole genome sequencing & analysis and identified four strains of A. faecalis. Furthermore, we characterized the genes in the genomic islands of A. faecalis isolates. We found four multidrug-resistant A. faecalis strains that showed resistance against vancomycin (MIC >1000 μg/ml), ceftazidime (50 μg/ml), colistin (50 μg/ml) and ciprofloxacin (50 μg/ml). From whole genome comparative analysis, we found more than 180 resistance genes compared to the reference sequence. Parts of our assembled contigs were found to be similar to different bacteria which included pbp1A and pbp2 imparting resistance to amoxicillin originally a part of Helicobacter and Bordetella pertussis. We also found the Mycobacterial insertion element IS6110 in the genomic islands of all four genomes. This prominent insertion element can be transferred and induce resistance to other bacterial genomes. The results thus are crucial in understanding the transfer of resistance genes in the environment and can help in developing regimes for antibiotic use in the food and poultry industry.
Two novel Gram-stain-negative, strictly-aerobic, rod-shaped (1.2 ± 3.4 μm × 0.3 ± 0.7 μm), and non-motile marine bacterial
species, designated MEBiC05379T
and MEBiC07777T,
were isolated from a marine sponge Pseudaxinella sp. in Gangneung
City and deep-sea sediments of the Ulleung basin in the East Sea of Korea, respectively. The 16S rRNA gene sequence
analysis revealed high levels of similarities between these strains and members of the genus Flavivirga (97.0–98.4% sequence
identities). Both novel strains revealed as mesophilic, neutrophilic in pH and slightly halophilic. Similar to those of other Flavivirga
members, the primary cellular fatty acids of both strains were iso-C15:0, iso-C15:1 G, iso-C15:03-OH, and iso-C17:0 3-OH,
with MEBiC05379T
and MEBiC07777T
containing relatively higher proportions of C12:
0 and summed feature 3 (
C16:1ω7c
and/or C16:
1ω6c). In both taxa, the major isoprenoid quinone was MK-6. The DNA G + C contents of MEBiC05379T
and
MEBiC07777T
genomes were 32.62 and 32.46 mol%, respectively. Compared to other members of Flavivirga, both strains
exhibited similar DNA G + C ratio and fatty acids pattern, yet enzyme expression and carbon sources utilization pattern were
different. Genomes of the genus Flavivirga showed enzyme preferences to fucoidan and sulfated galactans. Considering the
monophyly rule, AAI values delineate the genus Flavivirga from adjacent genera calculated to be 76.0–78.7%. Based on
the phenotypic, genomic and biochemical data, strains for MEBiC05379T
and MEBiC07777T
thus represent two novel species
in the genus Flavivirga, for which the names Flavivirga spongiicola sp. nov. (
MEBiC05379T [= KCTC 92527
T = JCM
16662
T]), and Flavivirga abyssicola sp. nov. (
MEBiC07777T [= KCTC 92563
T = JCM 36477
T]) are proposed.
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and has also been found in several places in Antarctica including lake, soil, and rocks. We performed genomic and transcriptomic
analyses of 5 strains from Antarctica and a type strain to understand their adaptation to different environments.
Interestingly, the isolates from rocks showed a low growth rate and smaller genome size than strains from the other isolation
sources (lake, soil, and groundwater). Based on these habitat-dependent characteristics, the strains could be classified
into two ecotypes, which showed differences in energy production, signal transduction, and transcription in the clusters of
orthologous groups of proteins (COGs) functional category. In addition, expression pattern changes revealed differences
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xylose degradation, depending on the nutritional status of their habitats. These findings provide crucial insights into the
environmental adaptation of bacteria, highlighting genetic diversity and regulatory mechanisms that enable them to thrive
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Norovirus (NoV) is the most common viral cause of acute gastroenteritis worldwide. Vitamin A has demonstrated the
potential to protect against gastrointestinal infections. However, the effects of vitamin A on human norovirus (HuNoV)
infections remain poorly understood. This study aimed to investigate how vitamin A administration affects NoV replication.
We demonstrated that treatment with retinol or retinoic acid (RA) inhibited NoV replication in vitro based on their effects
on HuNoV replicon-bearing cells and murine norovirus-1 (MNV-1) replication in murine cells. MNV replication in vitro
showed significant transcriptomic changes, which were partially reversed by retinol treatment. RNAi knockdown of CCL6,
a chemokine gene that was downregulated by MNV infection but upregulated by retinol administration, resulted in increased
MNV replication in vitro. This suggested a role of CCL6 in the host response to MNV infections. Similar gene expression
patterns were observed in the murine intestine after oral administration of RA and/or MNV-1.CW1. CCL6 directly decreased
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Candida species cause the most prevalent fungal illness, candidiasis.
Candida albicans is known to cause bloodstream infections.
This species is a commensal bacterium, but it can
cause hospital–acquired diseases, particularly in COVID-19
patients with impaired immune systems. Candida infections
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Coumarins are both naturally occurring and synthetically
produced. In this study, the biological activity of 40 coumarin
derivatives was used to create a three-dimensional quantitative
structure activity relationship (3D-QSAR) model. The
training and test minimum inhibitory concentration values
of C. albicans active compounds were split, and a regression
model based on statistical data was established. This model
served as a foundation for the creation of coumarin derivative
QSARs. This is a unique way to create new therapeutic compounds
for various ailments. We constructed novel structural
coumarin derivatives using the derived QSAR model, and the
models were confirmed using molecular docking and molecular
dynamics simulation.
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successful results in eliminating bacteria with high selective
toxicity. Bacteriophage encoded endolysin as an alternative
antibiotics is a peptidoglycan degrading enzyme with a low
rate of resistance. Here, the engineered endolysin was developed
to defeat multiple drug-resistant (MDR) Acinetobacter
baumannii. First, putative endolysin PA90 was predicted by
genome analysis of isolated Pseudomonas phage PBPA. The
His-tagged PA90 was purified from BL21(DE3) pLysS and
tested for the enzymatic activity using Gram-negative pathogens
known for having a high antibiotic resistance rate including
A. baumannii. Since the measured activity of PA90
was low, probably due to the outer membrane, cell-penetrating
peptide (CPP) DS4.3 was introduced at the N-terminus
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could protect waxworm from A. baumannii-induced death
by ~70% for ATCC 17978 or ~44% for MDR strain 1656-2
infection. Collectively, our data suggest that CPP-fused endolysin
can be an effective antibacterial agent against Gramnegative
pathogens regardless of antibiotics resistance mechanisms.
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CRC HCT116 (HCT116/5FUR) cells acquired butyrateinsensitive
properties. To date, the relationship between 5-
FU-resistant CRC and butyrate resistance has not been elucidated.
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resistance in HCT116/5FUR cells was strongly correlated
with the inhibition of the expression and function of
SMCT1, a major transporter of butyrate in colonocytes. L.
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strain (Δg511). Next, the biological characteristics of the
Δg511 mutant strain, including growth rate, conidia germination
rate, adaptation to environmental stresses, and nematocidal
activity, were compared with those of the wild-type
(WT) strain. The results showed that the JRL gene AOL_
s00083g511 did not affect fungal growth, conidia germination,
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prey on nematodes significantly. We speculate that this phenomenon
may be caused by a loss of the key β1–β2 loops in
the AOL_ s00083g511-encoded JRL domain and an intrinsic
genetic compensation of AOL_s00083g511 in this fungus.
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were similar; however, in the strong oxidation medium,
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than that of the WT strain, indicating that AOL_s00083g511
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provide a basis for further analysis of the related functions
of the JRL gene in A. oligospora and their potential roles
in the biological control of nematodes in the future.
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accumulating resistance genes on mobile genetic elements
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environments such as wastewater treatment plants and manured
soils have proven to be the major reservoirs and hotspots
of spreading antibiotic resistance genes (ARGs). As those
environments support the dissemination of MGEs through
the complex interactions that take place at the human-animalenvironment
interfaces, a growing One Health challenge is
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maintenance of ARGs in a bacterial chromosome and/or
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on bacterial fitness in the absence of antibiotics, and those
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environment interfaces. We also suggest alternative treatments
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Ogataea parapolymorpha (Hansenula polymorpha DL-1) is
a thermotolerant methylotrophic yeast with biotechnological
applications. Here, O. parapolymorpha genes whose expression
is induced in response to heat shock were identified by
transcriptome analysis and shown to possess heat shock elements
(HSEs) in their promoters. The function of O. parapolymorpha
HSF1 encoding a putative heat shock transcription
factor 1 (OpHsf1) was characterized in the context of heat
stress response. Despite exhibiting low sequence identity
(26%) to its Saccharomyces cerevisiae homolog, OpHsf1 harbors
conserved domains including a DNA binding domain
(DBD), domains involved in trimerization (TRI), transcriptional
activation (AR1, AR2), transcriptional repression (CE2),
and a C-terminal modulator (CTM) domain. OpHSF1 could
complement the temperature sensitive (Ts) phenotype of a
S. cerevisiae hsf1 mutant. An O. parapolymorpha strain with
an H221R mutation in the DBD domain of OpHsf1 exhibited
significantly retarded growth and a Ts phenotype. Intriguingly,
the expression of heat-shock-protein‒coding genes harboring
HSEs was significantly decreased in the H221R mutant
strain, even under non-stress conditions, indicating the importance
of the DBD for the basal growth of O. parapolymorpha.
Notably, even though the deletion of C-terminal domains
(ΔCE2, ΔAR2, ΔCTM) of OpHsf1 destroyed complementation
of the growth defect of the S. cerevisiae hsf1 strain,
the C-terminal domains were shown to be dispensable in O.
parapolymorpha. Overexpression of OpHsf1 in S. cerevisiae
increased resistance to transient heat shock, supporting the
idea that OpHsf1 could be useful in the development of heatshock‒
resistant yeast host strains.
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Multi-omics approaches, including metagenomics and single-
cell amplified genomics, have revolutionized our understanding
of the hidden diversity and function of microbes
in nature. Even in the omics age, cultivation is an essential
discipline in microbial ecology since microbial cultures are
necessary to assess the validity of an in silico prediction about
the microbial metabolism and to isolate viruses infecting bacteria
and archaea. However, the ecophysiological characteristics
of predominant freshwater bacterial lineages remain
largely unknown due to the scarcity of cultured representatives.
In an ongoing effort to cultivate the uncultured majority
of freshwater bacteria, the most abundant freshwater
Actinobacteria acI clade has recently been cultivated from
Lake Soyang through catalase-supplemented high-throughput
cultivation based on dilution-to-extinction. This method
involves physical isolation of target microbes from mixed populations,
culture media simulating natural habitats, and removal
of toxic compounds. In this protocol, we describe detailed
procedures for isolating freshwater oligotrophic microbes,
as well as the essence of the dilution-to-extinction culturing.
As a case study employing the catalase-supplemented
dilution-to-extinction protocol, we also report a cultivation
trial using a water sample collected from Lake Soyang. Of the
480 cultivation wells inoculated with a single lake-water sample,
75 new acI strains belonging to 8 acI tribes (acI-A1, A2,
A4, A5, A6, A7, B1, B4, C1, and C2) were cultivated, and each
representative strain per subclade could be revived from glycerol
stocks. These cultivation results demonstrate that the
protocol described in this study is efficient in isolating freshwater
bacterioplankton harboring streamlined genomes.
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Helicobacter pylori, present in the stomach lining, is a Gramnegative
bacterium that causes various gastrointestinal diseases,
including gastritis and peptic ulcers. Propolis is a natural
resinous substance collected from a variety of plants,
and contains several natural bioactive substances. The aim of
this study was to investigate the anti-inflammatory and antioxidative
effects of Korean propolis on H. pylori-induced damage
in the human adenocarcinoma gastric cell line. The propolis
used in this study was obtained from the Korea Beekeeping
Association in South Korea. The expression of pro-inflammatory
interleukins (ILs), such as IL-8, IL-12, IL-1β, tumor
necrosis factor alpha, cyclooxygenase-2, and inducible
nitric oxide synthase, which was increased after H. pylori infection,
significantly decreased in a dose-dependent manner
upon pretreatment with Korean propolis, because of the suppression
of mitogen-activated protein kinases and nuclear
factor κB pathway. The anti-oxidative activity of propolis was
assessed using the 2,2-diphenyl-1-picrylhydrazyl hydrate free
radical assay. Korean propolis showed significant anti-oxidative
effects via reactive oxygen species scavenging. In addition,
pretreatment with Korean propolis upregulated the
expression of anti-oxidant enzymes through Nrf2 signaling
activation. These findings indicate that the use of Korean propolis,
which has anti-inflammatory and anti-oxidative effects,
can be promising for the prevention of H. pylori-induced gastric
damage.
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The saprophytic fungus Trichoderma reesei has long been used
as a model to study microbial degradation of lignocellulosic
biomass. The major cellulolytic enzymes of T. reesei are the
cellobiohydrolases CBH1 and CBH2, which constitute more
than 70% of total proteins secreted by the fungus. However,
their physiological functions and effects on enzymatic hydrolysis
of cellulose substrates are not sufficiently elucidated.
Here, the cellobiohydrolase-encoding genes cbh1 and cbh2
were deleted, individually or combinatively, by using an auxotrophic
marker-recycling technique in T. reesei. When cultured
on media with different soluble carbon sources, all three
deletion strains (Δcbh1, Δcbh2, and Δcbh1Δcbh2) exhibited
no dramatic variation in morphological phenotypes, but their
growth rates increased apparently when cultured on soluble
cellulase-inducing carbon sources. In addition, Δcbh1 showed
dramatically reduced growth and Δcbh1Δcbh2 could hardly
grew on microcrystalline cellulose (MCC), whereas all strains
grew equally on sodium carboxymethyl cellulose (CMC-Na),
suggesting that the influence of the CBHs on growth was carbon
source-dependent. Moreover, five representative cellulose
substrates were used to analyse the influence of the absence
of CBHs on saccharification efficiency. CBH1 deficiency
significantly affected the enzymatic hydrolysis rates of various
cellulose substrates, where acid pre-treated corn stover
(PCS) was influenced the least. CBH2 deficiency reduced the
hydrolysis of MCC, PCS, and acid pre-treated and delignified
corncob but improved the hydrolysis ability of filter paper.
These results demonstrate the specific contributions of
CBHs to the hydrolysis of different types of biomass, which
could facilitate the development of tailor-made strains with
highly efficient hydrolysis enzymes for certain biomass types
in the biofuel industry.
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In this study, we investigated the effect of cryptic prophage
regions in a blaNDM-1-bearing plasmid, which was identified in
a patient from South Korea, on the survival of bacteria against
adverse environmental conditions. First, we conjugated the
intact plasmid and plasmids with deleted cryptic prophages
into Escherichia coli DH5α. The E. coli transconjugants carrying
the plasmid with intact cryptic prophages showed increased
survival during treatment with a high concentration
of NaCl, high and low temperatures, an oxidative stressor
(H2O2), and an immunological stressor (human serum). By
contrast, the transconjugants carrying the plasmid with a
single-cryptic prophage knockout did not show any change
in survival rates. mRNA expression analyses revealed that the
genes encoding sigma factor proteins were highly upregulated
by the tested stressors and affected the expression of
various proteins (antioxidant, cell osmosis-related, heat shock,
cold shock, and universal stress proteins) associated with the
specific defense against each stress. These findings indicate
that a bacterial strain carrying a plasmid with intact carbapenemase
gene and cryptic prophage regions exhibited an increased
resistance against simulated environmental stresses,
and cryptic prophages in the plasmid might contribute to this
enhanced stress resistance. Our study indicated that the coselection
of antibiotic resistance and resistance to other stresses
may help bacteria to increase survival rates against adverse
environments and disseminate.
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α-Glucosidase is a crucial enzyme for the production of isomaltooligosaccharide.
In this study, a novel method comprising
eosin Y (EY) and α-D-methylglucoside (AMG) in glass
plates was tested for the primary screening of α-glucosidaseproducing
strains. First, α-glucosidase-producing Aspergillus
niger strains were selected on plates containing EY and AMG
based on transparent zone formation resulting from the solubilization
of EY by the hydrolyzed product. Conventional methods that use trypan blue (TB) and p-nitrophenyl-α-Dglucopyranoside
(pPNP) as indicators were then compared
with the new strategy. The results showed that EY-containing
plates provide the advantages of low price and higher specificity
for the screening of α-glucosidase-producing strains.
We then evaluated the correlation between the hydrolytic activity
of α-glucosidase and diffusion distance, and found that
good linearity could be established within a 6–75 U/ml enzyme
concentration range. Finally, the hydrolytic and transglycosylation
activities of α-glucosidase obtained from the
target isolates were determined by EY plate assay and 3,5-
dinitrosalicylic acid-Saccharomyces cerevisiae assay, respectively.
The results showed that the diameter of the transparent
zone varied among isolates was positively correlated with
α-glucosidase hydrolytic activity, while good linearity could
also be established between α-glucosidase transglycosylation
activity and non-fermentable reducing sugars content. With
this strategy, 7 Aspergillus niger mutants with high yield of
α-glucosidase from 200 obvious single colonies on the primary
screen plate were obtained.
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