Antibiotics have long been used for anti-infective control of
bacterial infections, growth promotion in husbandry, and
prophylactic protection against plant pathogens. However,
their inappropriate use results in the emergence and spread
of multiple drug resistance (MDR) especially among various
bacterial populations, which limits further administration
of conventional antibiotics. Therefore, the demand for novel
anti-infective approaches against MDR diseases becomes
increasing in recent years. The peptide nucleic acid (PNA)-
based technology has been proposed as one of novel antiinfective
and/or therapeutic strategies. By definition, PNA
is an artificially synthesized nucleic acid mimic structurally
similar to DNA or RNA in nature and linked one another via
an unnatural pseudo-peptide backbone, rendering to its stability
in diverse host conditions. It can bind DNA or RNA
strands complimentarily with high affinity and sequence specificity,
which induces the target-specific gene silencing by
inhibiting transcription and/or translation. Based on these
unique properties, PNA has been widely applied for molecular
diagnosis as well as considered as a potential anti-infective
agent. In this review, we discuss the general features
of PNAs and their application to various bacterial pathogens
as new anti-infective or antimicrobial agents.
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Sortases are cysteine transpeptidases that assemble surface
proteins and pili in their cell envelope. Encoded by all Grampositive
bacteria, few Gram-negative bacteria and archaea,
sortases are currently divided into six classes (A-F). Due to
the steep increase in bacterial genome data in recent years,
the number of sortase homologues have also escalated rapidly.
In this study, we used protein sequence similarity networks
to explore the taxonomic diversity of sortases and also to evaluate
the current classification of these enzymes. The resultant
data suggest that sortase classes A, B, and D predominate in
Firmicutes and classes E and F are enriched in Actinobacteria,
whereas class C is distributed in both Firmicutes and Actinobacteria
except Streptomyces family. Sortases were also observed
in various Gram-negatives and euryarchaeota, which
should be recognized as novel classes of sortases. Motif analysis
around the catalytic cysteine was also performed and
suggested that the residue at 2nd position from cysteine may
help distinguish various sortase classes. Moreover, the sequence
analysis indicated that the catalytic arginine is highly
conserved in almost all classes except sortase F in which arginine
is replaced by asparagine in Actinobacteria. Additionally,
class A sortases showed higher structural variation as compared
to other sortases, whereas inter-class comparisons suggested
structures of class C and D2 exhibited best similarities.
A better understanding of the residues highlighted in
this study should be helpful in elucidating their roles in substrate
binding and the sortase function, and successively could
help in the development of strong sortase inhibitors.
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A Gram-stain-negative and facultatively aerobic bacterium,
designated as strain CBA4604T, was isolated from a traditional
Korean salted and fermented shrimp food (saeu-jeot).
Phylogenetic analysis based on 16S rRNA gene sequences
showed that strain CBA4604T formed a clearly distinct phyletic
lineage from closely related species within the genus
Paracoccus. Strain CBA4604T was the most closely related to
P. koreensis Ch05T (97.5% 16S rRNA gene sequence similarity)
and other type strains (≤ 97.0%). The genome comprised a
chromosome and two plasmids of 3,299,166 bp with 66.5%
G+C content. The DNA-DNA relatedness values between
strain CBA4604T and P. koreensis Ch05T, P. alcaliphilus DSM
8512T, and P. stylophorae KTW-16T were 30.5%, 22.9%, and
16.7%, respectively. Cells of the strain were short rod-shaped
and oxidase- and catalase-positive. The growth of strain CBA-
4604T was observed at 10–40°C (optimum, 37°C), pH 6.0–10.0
(optimum, pH 7.0), and in the presence of 0–8.0% (w/v) NaCl
(optimum, 0–2.0%). Strain CBA4604T contained ubiquinone
10 as the sole isoprenoid quinone and summed feature 8 (C18:1
ω7c/C18:1 ω6c) and C18:0 as the major cellular fatty acids. The
polar lipids consisted of phosphatidylcholine, phosphatidylglycerol,
diphosphatidylglycerol, phospholipid, an unidentified
aminolipid, an unidentified glycolipid, and three unidentified
lipids. Based on its phylogenetic, genomic, phenotypic,
and chemotaxonomic features, we concluded that strain CBA-
4604T represents a novel species in the genus Paracoccus and
we propose the name Paracoccus jeotgali sp. nov. The type
strain is CBA4604T (= KACC 19579T = JCM 32510T).
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Next-generation DNA sequencing technology was applied
to generate molecular data from semiarid reservoirs during
well-defined seasons. Target sequences of 16S-23S rRNA ITS
and cpcBA-IGS were used to reveal the taxonomic groups of
cyanobacteria present in the samples, and genes coding for
cyanotoxins such as microcystins (mcyE), saxitoxins (sxtA),
and cylindrospermopsins (cyrJ) were investigated. The presence
of saxitoxins in the environmental samples was evaluated
using ELISA kit. Taxonomic analyses of high-throughput
DNA sequencing data showed the dominance of the genus
Microcystis in Mundaú reservoir. Furthermore, it was the
most abundant genus in the dry season in Ingazeira reservoir.
In the rainy season, 16S-23S rRNA ITS analysis revealed that
Cylindrospermopsis raciborskii comprised 46.8% of the cyanobacterial
community in Ingazeira reservoir, while the cpcBAIGS
region revealed that C. raciborskii (31.8%) was the most
abundant taxon followed by Sphaerospermopsis aphanizomenoides
(17.3%) and Planktothrix zahidii (16.6%). Despite
the presence of other potential toxin-producing genera, the
detected sxtA gene belonged to C. raciborskii, while the mcyE
gene belonged to Microcystis in both reservoirs. The detected
mcyE gene had good correlation with MC content, while the
amplification of the sxtA gene was related to the presence of
STX. The cyrJ gene was not detected in these samples. Using
DNA analyses, our results showed that the cyanobacterial
composition of Mundaú reservoir was similar in successive
dry seasons, and it varied between seasons in Ingazeira reservoir.
In addition, our data suggest that some biases of analysis
influenced the cyanobacterial communities seen in
the NGS output of Ingazeira reservoir.
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Microbial community assembly is affected by trade-offs between
deterministic and stochastic processes. However, the
mechanisms underlying the relative influences of the two
processes remain elusive. This knowledge gap limits our ability
to understand the effects of community assembly processes
on microbial community structures and functions. To better
understand community assembly mechanisms, the community
dynamics of bacterial ecological groups were investigated
based on niche breadths in 23 soil plots from subalpine coniferous
forests on the Loess Plateau in Shanxi, China. Here,
the overall community was divided into the ecological groups
that corresponded to habitat generalists, ‘other taxa’ and specialists.
Redundancy analysis based on Bray-Curtis distances
(db-RDA) and multiple regression tree (MRT) analysis indicated
that soil organic carbon (SOC) was a general descriptor
that encompassed the environmental gradients by which the
communities responded to, because it can explain more significant
variations in community diversity patterns. The three
ecological groups exhibited different niche optima and degrees
of specialization (i.e., niche breadths) along the SOC
gradient, suggesting the presence of a gradient in tolerance
for environmental heterogeneity. The inferred community
assembly processes varied along the SOC gradient, wherein
a transition was observed from homogenizing dispersal to
variable selection that reflects increasing deterministic processes.
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samples. The methanogenic community in YR samples was
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methanogens suggested that the methanogenic community
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To develop a starter culture system for improving the shelflife
and quality of kimchi, we prepared a mixed starter composed
of Lactococcus lactis and Leuconostoc citreum. Two
strains, L. lactis WiKim0098 and Leu. citreum WiKim0096,
showed high antimicrobial activity and mannitol productivity,
respectively. These lactic acid bacteria (LAB) were introduced
as a starter into kimchi following cultivation in foodgrade
liquid medium. Two kimchi samples, with and without
starter, were fermented for 12 days at 10°C. Compared to the
control kimchi without starter, a lower initial pH and higher
number of LAB were observed in kimchi with starter at 0 day.
However, the starter in kimchi prolonged the period taken by
kimchi to reach to pH 4.2 by approximately 1.5-fold compared
to that in the control kimchi. To estimate the effect of
the starter on the flavor of kimchi, metabolite changes were
evaluated by gas chromatography/mass spectrometry. In starter
fermented kimchi, the levels of mannitol and amino acid,
which are associated with the flavor of kimchi, were increased
following fermentation. The amount of mannitol was confirmed
by high-performance liquid chromatography analysis,
showing concentrations of 3.4 and 5.1 mg/ml for the control
and starter fermented kimchi, respectively. Thus, mixed starter
inoculated with L. lactis WiKim0098 and Leu. citreum
WiKim0096 may extend the shelf-life of kimchi and improve
its sensory characteristics.
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Fusarium solani has drawn phytopathogenic, biotechnological,
and medical interest. In humans, it is associated with
localized infections, such as onychomycosis and keratomycosis,
as well as invasive infections in immunocompromised
patients. One pathogenicity factor of filamentous fungi is biofilm
formation. There is still only scarce information about
the in vitro mechanism of the formation and composition of
F. solani biofilm. In this work, we describe the biofilm formed
by a clinical keratomycosis isolate in terms of its development,
composition and susceptibility to different antifungals and
ultraviolet light (UV) at different biofilm formation stages.
We found five biofilm formation stages using scanning electron
microscopy: adherence, germination, hyphal development,
maturation, and cell detachment. Using epifluorescence
microscopy with specific fluorochromes, it was elucidated
that the extracellular matrix consists of carbohydrates, proteins,
and extracellular DNA. Specific inhibitors for these
molecules showed significant biofilm reductions. The antifungal
susceptibility against natamycin, voriconazole, caspofungin,
and amphotericin B was evaluated by metabolic activity
and crystal violet assay, with the F. solani biofilm preformation
to 24 h increased in resistance to natamycin, voriconazole,
and caspofungin, while the biofilm preformation
to 48 h increased in resistance to amphotericin B. The preformed
biofilm at 24 h protected and reduced UV light
mortality. F. solani isolate could produce a highly structured
extra biofilm; its cellular matrix consists of carbohydrate polymers,
proteins, and eDNA. Biofilm confers antifungal resistance
and decreases its susceptibility to UV light. The fungal
biofilm functions as a survival strategy against antifungals
and environmental factors.
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Federica De Lise , Francesca Mensitieri , Giulia Rusciano , Fabrizio Dal Piaz , Giovanni Forte , Flaviana Di Lorenzo , Antonio Molinaro , Armando Zarrelli , Valeria Romanucci , Valeria Cafaro , Antonio Sasso , Amelia Filippelli , Alberto Di Donato , Viviana Izzo
J. Microbiol. 2019;57(6):498-508. Published online May 27, 2019
Outer membrane vesicles (OMVs) are nanostructures of 20–
200 nm diameter deriving from the surface of several Gramnegative
bacteria. OMVs are emerging as shuttles involved in
several mechanisms of communication and environmental
adaptation. In this work, OMVs were isolated and characterized
from Novosphingobium sp. PP1Y, a Gram-negative
non-pathogenic microorganism lacking LPS on the outer
membrane surface and whose genome was sequenced and
annotated. Scanning electron microscopy performed on samples
obtained from a culture in minimal medium highlighted
the presence of PP1Y cells embedded in an extracellular matrix
rich in vesicular structures. OMVs were collected from
the exhausted growth medium during the mid-exponential
phase, and purified by ultracentrifugation on a sucrose gradient.
Atomic force microscopy, dynamic light scattering and
nanoparticle tracking analysis showed that purified PP1Y
OMVs had a spherical morphology with a diameter of ca. 150
nm and were homogenous in size and shape. Moreover, proteomic
and fatty acid analysis of purified OMVs revealed a
specific biochemical “fingerprint”, suggesting interesting details
concerning their biogenesis and physiological role. Moreover,
these extracellular nanostructures do not appear to be
cytotoxic on HaCaT cell line, thus paving the way to their
future use as novel drug delivery systems.
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J. Microbiol. 2019;57(6):509-520. Published online May 27, 2019
Mucor circinelloides is a pathogenic fungus and etiologic agent
of mucormycosis. In 2013, cases of gastrointestinal illness
after yogurt consumption were reported to the US FDA, and
the producer found that its products were contaminated with
Mucor. A previous study found that the Mucor strain isolated
from an open contaminated yogurt exhibited virulence
in a murine systemic infection model and showed that this
strain is capable of surviving passage through the gastrointestinal
tract of mice. In this study, we isolated another Mucor
strain from an unopened yogurt that is closely related but
distinct from the first Mucor strain and subsequently examined
if Mucor alters the gut microbiota in a murine host
model. DNA extracted from a ten-day course of stool samples
was used to analyze the microbiota in the gastrointestinal
tracts of mice exposed via ingestion of Mucor spores. The
bacterial 16S rRNA gene and fungal ITS1 sequences obtained
were used to identify taxa of each kingdom. Linear regressions
revealed that there are changes in bacterial and fungal abundance
in the gastrointestinal tracts of mice which ingested
Mucor. Furthermore, we found an increased abundance of
the bacterial genus Bacteroides and a decreased abundance
of the bacteria Akkermansia muciniphila in the gastrointestinal
tracts of exposed mice. Measurements of abundances
show shifts in relative levels of multiple bacterial and fungal
taxa between mouse groups. These findings suggest that exposure
of the gastrointestinal tract to Mucor can alter the microbiota
and, more importantly, illustrate an interaction between
the intestinal mycobiota and bacteriota. In addition, Mucor was able to induce increased permeability in epithelial
cell monolayers in vitro, which might be indicative of unstable
intestinal barriers. Understanding how the gut microbiota is
shaped is important to understand the basis of potential methods
of treatment for gastrointestinal illness. How the gut
microbiota changes in response to exposure, even by pathogens
not considered to be causative agents of food-borne illness,
may be important to how commercial food producers
prevent and respond to contamination of products aimed at
the public. This study provides evidence that the fungal microbiota,
though understudied, may play an important role
in diseases of the human gut.
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Characteristics of naringinase nano-encapsulated forms on
different carrier materials (chitosan and alginate polymers)
were investigated in this study. Screening of twelve fungal isolates
for naringinase production indicated that Trichoderma
longibrachiatum was the most promising. Grapefruit rind was
used as a substrate containing naringin for naringinase production.
TEM micrographs showed that chitosan nano-capsules
were applied for the production of morphologically homogeneous
enzymatic nano-particles with high enzyme encapsulation
efficiency, small asymmetric sizes (from 15.09 to
27.07 nm with the mean of 21.8 nm) and rough surfaces compared
to nano-encapsulated naringinase in alginate which
showed nano-particle size (from 33.37 to 51.01 nm with the
mean of 43.03 nm). It was revealed that the highest naringinase
activity was found in case of chitosan nano-capsule naringinase
compared to alginate nano-capsule one. Thermogram
analysis (TGA) showed that the free enzyme loses about
92% of its weight at approximately 110°C, while the nanoencapsulated
ones show more stability at higher temperatures.
Conclusively, the nano-capsulation process improves the kinetics
and operational stability so could be useful as a debittering
agent for various thermal processing applications in
citrus juices industries which makes the fruit juice more acceptable
and cost-effective to the consumer.
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Bacteriophage therapy was an ascendant technology for combating
bacterial infections before the golden age of antibiotics,
but the therapeutic potential of phages was largely ignored
after the discovery of penicillin. Recently, with antibioticresistant
infections on the rise, these phages are receiving renewed
attention to combat problematic bacterial infections.
Our approach is to enhance bacteriophages with antimicrobial
peptides, short peptides with broad-spectrum antibiotic or
antibiofilm effects. We inserted coding sequences for 1018,
an antimicrobial peptide previously shown to be an effective
broad-spectrum antimicrobial and antibiofilm agent, or the
fluorescent marker mCherry, into the T7Select phage genome.
Transcription and production of 1018 or mCherry began
rapidly after E. coli cultures were infected with genetically modified
phages. mCherry fluorescence, which requires a 90 min
initial maturation period, was observed in infected cultures
after 2 h of infection. Finally, we tested phages expressing 1018
(1018 T7) against bacterial planktonic cultures and biofilms,
and found the 1018 T7 phage was more effective than the
unmodified T7Select phage at both killing planktonic cells and
eradicating established biofilms, validating our phage-driven
antimicrobial peptide expression system. The combination
of narrow-spectrum phages delivering relatively high local
doses of broad-spectrum antimicrobials could be a powerful method to combat resistant infections. The experiments we
describe prove this combination is feasible in vitro, but further
testing and optimization are required before genetically modified
phages are ready for use in vivo.
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In the article by Lee et al. published in Journal of Microbiology 2016; 54, 809–813, The KCTC accession number KCTC 18866T
in abstract and foot note should be corrected to KCTC 33839T.
The sentence in abstract should have read: Based on the phylogenetic, phenotypic, and chemotaxonomic data, strain 16MFT21T
(=KCTC 33839T =JCM 31664T).
The species description in foot note should have read: The NCBI GenBank/EMBL/DDBJ accession number for the 16S rRNA
gene sequence of strain 16MFT21T (=KCTC 33839T =JCM 31664T) is KX753358.
We apologize for any inconvenience that this may have caused.