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- Application of fast expectation-maximization microbial source tracking to discern fecal contamination in rivers exposed to low fecal inputs
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Youfen Xu , Ganghua Han , Hongxun Zhang , Zhisheng Yu , Ruyin Liu
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J. Microbiol. 2022;60(6):594-601. Published online April 18, 2022
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DOI: https://doi.org/10.1007/s12275-022-1651-9
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Abstract
- Community-based microbial source tracking (MST) can be
used to determine fecal contamination from multiple sources
in the aquatic environment. However, there is little scientific
information on its application potential in water environmental
management. Here, we compared SourceTracker and
Fast Expectation-maximization Microbial Source Tracking
(FEAST) performances on environmental water bodies exposed
to low fecal pollution and evaluated treatment effects
of fecal pollution in the watershed utilizing community-based
MST. Our results showed that FEAST overall outperformed
SourceTracker in sensitivity and stability, and was able to discern
multi-source fecal contamination (mainly chicken feces)
in ambient water bodies exposed to low fecal inputs. Consistent
with our previous PCR/qPCR-based MST assays, FEAST
analysis indicates that fecal pollution has been significantly
mitigated through comprehensive environmental treatment
by the local government. This study suggests that FEAST can
be a powerful tool for accurately evaluating the contribution
of multi-source fecal contamination in environmental water,
facilitating environmental management.
- Regulator of ribonuclease activity modulates the pathogenicity of Vibrio vulnificus
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Jaejin Lee , Eunkyoung Shin , Jaeyeong Park , Minho Lee , Kangseok Lee
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J. Microbiol. 2021;59(12):1133-1141. Published online November 9, 2021
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DOI: https://doi.org/10.1007/s12275-021-1518-5
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Abstract
- RraA, a protein regulator of RNase E activity, plays a unique
role in modulating the mRNA abundance in Escherichia coli.
The marine pathogenic bacterium Vibrio vulnificus also possesses
homologs of RNase E (VvRNase E) and RraA (VvRraA1
and VvRraA2). However, their physiological roles have not
yet been investigated. In this study, we demonstrated that
VvRraA1 expression levels affect the pathogenicity of V. vulnificus.
Compared to the wild-type strain, the VvrraA1-deleted
strain (ΔVvrraA1) showed decreased motility, invasiveness,
biofilm formation ability as well as virulence in mice; these
phenotypic changes of ΔVvrraA1 were restored by the exogenous
expression of VvrraA1. Transcriptomic analysis indicated
that VvRraA1 expression levels affect the abundance
of a large number of mRNA species. Among them, the halflives
of mRNA species encoding virulence factors (e.g., smcR
and htpG) that have been previously shown to affect VvrraA1
expression-dependent phenotypes were positively correlated
with VvrraA1 expression levels. These findings suggest that
VvRraA1 modulates the pathogenicity of V. vulnificus by regulating
the abundance of a subset of mRNA species.
- iTRAQ-facilitated proteomic analysis of Bacillus cereus via degradation of malachite green
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Bobo Wang , Jing Lu , Junfang Zheng , Zhisheng Yu
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J. Microbiol. 2021;59(2):142-150. Published online February 1, 2021
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DOI: https://doi.org/10.1007/s12275-021-0441-0
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Abstract
- The wide use of malachite green (MG) as a dye has caused
substantial concern owing to its toxicity. Bacillus cereus can
against the toxic effect of MG and efficiently decolourise it.
However, detailed information regarding its underlying adaptation
and degradation mechanisms based on proteomic
data is scarce. In this study, the isobaric tags for relative and
absolute quantitation (iTRAQ)-facilitated quantitative method
was applied to analyse the molecular mechanisms by
which B. cereus degrades MG. Based on this analysis, 209
upregulated proteins and 198 downregulated proteins were
identified with a false discovery rate of 1% or less during MG
biodegradation. Gene ontology and KEGG analysis determined
that the differentially expressed proteins were enriched
in metabolic processes, catalytic activity, antioxidant activity,
and responses to stimuli. Furthermore, real-time qPCR was
utilised to further confirm the regulated proteins involved
in benzoate degradation. The proteins BCE_4076 (Acetyl-CoA
acetyltransferase), BCE_5143 (Acetyl-CoA acetyltransferase),
BCE_5144 (3-hydroxyacyl-CoA dehydrogenase), BCE_4651
(Enoyl-CoA hydratase), and BCE_5474 (3-hydroxyacyl-CoA
dehydrogenase) involved in the benzoate degradation pathway
may play an important role in the biodegradation of MG
by B. cereus. The results of this study not only provide a comprehensive
view of proteomic changes in B. cereus upon MG
loading but also shed light on the mechanism underlying
MG biodegradation by B. cereus.
- The putative polysaccharide synthase AfCps1 regulates Aspergillus fumigatus morphogenesis and conidia immune response in mouse bone marrow-derived macrophages
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Sha Wang , Anjie Yuan , Liping Zeng , Sikai Hou , Meng Wang , Lei Li , Zhendong Cai , Guowei Zhong
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J. Microbiol. 2021;59(1):64-75. Published online November 17, 2020
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DOI: https://doi.org/10.1007/s12275-021-0347-x
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Abstract
- Aspergillus fumigatus is a well-known opportunistic pathogen
that causes invasive aspergillosis (IA) infections with high
mortality in immunosuppressed individuals. Morphogenesis,
including hyphal growth, conidiation, and cell wall biosynthesis
is crucial in A. fumigatus pathogenesis. Based on a previous
random insertional mutagenesis library, we identified
the putative polysaccharide synthase gene Afcps1 and its paralog
Afcps2. Homologs of the cps gene are commonly found
in the genomes of most fungal and some bacterial pathogens.
Afcps1/cpsA is important in sporulation, cell wall composition,
and virulence. However, the precise regulation patterns
of cell wall integrity by Afcps1/cpsA and further effects on the
immune response are poorly understood. Specifically, our
in-depth study revealed that Afcps1 affects cell-wall stability,
showing an increased resistance of ΔAfcps1 to the chitinmicrofibril
destabilizing compound calcofluor white (CFW)
and susceptibility of ΔAfcps1 to the β-(1,3)-glucan synthase
inhibitor echinocandin caspofungin (CS). Additionally, deletion
of Afcps2 had a normal sporulation phenotype but
caused hypersensitivity to Na+ stress, CFW, and Congo red
(CR). Specifically, quantitative analysis of cell wall composition
using high-performance anion exchange chromatography-
pulsed amperometric detector (HPAEC-PAD) analysis
revealed that depletion of Afcps1 reduced cell wall glucan
and chitin contents, which was consistent with the downregulation
of expression of the corresponding biosynthesis
genes. Moreover, an elevated immune response stimulated
by conidia of the ΔAfcps1 mutant in marrow-derived macrophages
(BMMs) during phagocytosis was observed. Thus,
our study provided new insights into the function of polysaccharide
synthase Cps1, which is necessary for the maintenance
of cell wall stability and the adaptation of conidia to
the immune response of macrophages in A. fumigatus.
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