Severe fever with thrombocytopenia syndrome (SFTS) is a life-threatening viral zoonosis. The causative agent of this disease is the Dabie bandavirus, which is usually known as the SFTS virus (SFTSV). Although the role of vertebrates in SFTSV transmission to humans remains uncertain, some reports have suggested that dogs could potentially transmit SFTSV to humans. Consequently, preventive measures against SFTSV in dogs are urgently needed. In the present study, dogs were immunized three times at two-week intervals with formaldehyde-inactivated SFTSV with two types of adjuvants. SFTSV (KCD46) was injected into all dogs two weeks after the final immunization. Control dogs showed viremia from 2 to 4 days post infection (dpi), and displayed white pulp atrophy in the spleen, along with a high level of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling assay (TUNEL) positive area. However, the inactivated SFTSV vaccine groups exhibited rare pathological changes and significantly reduced TUNEL positive areas in the spleen. Furthermore, SFTSV viral loads were not detected at any of the tested dpi. Our results indicate that both adjuvants can be safely used in combination with an inactivated SFTSV formulation to induce strong neutralizing antibodies. Inactivated SFTSV vaccines effectively prevent pathogenicity and viremia in dogs infected with SFTSV. In conclusion, our study highlighted the potential of inactivated SFTSV vaccination for SFTSV control in dogs.
Viridiana Alejandre-Castañeda , J. Alberto Patiño-Medina , Marco I. Valle-Maldonado , Alexis García , Rafael Ortiz-Alvarado , León F. Ruíz-Herrera , Karla Viridiana Castro-Cerritos , Joel Ramírez-Emiliano , Martha I. Ramírez-Díaz , Victoriano Garre , Soo Chan Lee , Víctor Meza-Carmen
J. Microbiol. 2023;61(12):1043-1062. Published online December 19, 2023
Mucormycosis is a lethal and difficult-to-treat fungal infection caused by fungi of the order Mucorales. Mucor lusitanicus,
a member of Mucorales, is commonly used as a model to understand disease pathogenesis. However, transcriptional control
of hyphal growth and virulence in Mucorales is poorly understood. This study aimed to investigate the role of Tec proteins,
which belong to the TEA/ATTS transcription factor family, in the hyphal development and virulence of M. lusitanicus. Unlike
in the genome of Ascomycetes and Basidiomycetes, which have a single Tec homologue, in the genome of Mucorales, two
Tec homologues, Tec1 and Tec2, were found, except in that of Phycomyces blakesleeanus, with only one Tec homologue.
tec1 and tec2 overexpression in M. lusitanicus increased mycelial growth, mitochondrial content and activity, expression of
the rhizoferrin synthetase-encoding gene rfs, and virulence in nematodes and wax moth larvae but decreased cAMP levels
and protein kinase A (PKA) activity. Furthermore, tec1- and tec2-overexpressing strains required adequate mitochondrial
metabolism to promote the virulent phenotype. The heterotrimeric G beta subunit 1-encoding gene deletant strain (Δgpb1)
increased cAMP-PKA activity, downregulation of both tec genes, decreased both virulence and hyphal development, but tec1
and tec2 overexpression restored these defects. Overexpression of allele-mutated variants of Tec1(S332A) and Tec2(S168A)
in the putative phosphorylation sites for PKA increased both virulence and hyphal growth of Δgpb1. These findings suggest
that Tec homologues promote mycelial development and virulence by enhancing mitochondrial metabolism and rhizoferrin
accumulation, providing new information for the rational control of the virulent phenotype of M. lusitanicus.
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a wide range of pathogens, making faecal contamination a leading source of waterborne diseases around the world. Yet
monitoring bacteria or viruses in aquatic environments is time consuming and expensive. The standard indicators of faecal
pollution all have limitations, including difficulty in determining the source due to lack of host specificity, poor connection
with the presence of non-bacterial pathogens, or low environmental persistence. Innovative monitoring techniques are sorely
needed to provide more accurate and targeted solutions. Viruses are a promising alternative to faecal indicator bacteria for
monitoring, as they are more persistent in ambient water, more abundant in faeces, and are extremely host-specific. Given
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several are of interest. In parallel, the ongoing development of molecular techniques coupled with metagenomics and bioinformatics
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of faecal contamination monitoring with the following aims (i) to identify the characteristics of the main viral indicators of
faecal contamination, including human enteric viruses, bacteriophages, CRESS and plant viruses, (ii) to assess how these
have been used to monitor water pollution in recent years, (iii) to evaluate the reliability of recent detection methods of such
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The mammalian intestinal tract contains trillions of bacteria.
However, the genetic factors that allow gut symbiotic bacteria
to occupy intestinal niches remain poorly understood. Here,
we identified genetic determinants required for Bacteroides
thetaiotaomicron colonization in the gut using transposon
sequencing analysis. Transposon insertion in BT2391, which
encodes a hybrid two-component system, increased the competitive
fitness of B. thetaiotaomicron. The BT2391 mutant
showed a growth advantage in a mucin-dependent manner
and had an increased ability to adhere to mucus-producing
cell lines. The increased competitive advantage of the BT2391
mutant was dependent on the BT2392–2395 locus containing
susCD homologs. Deletion of BT2391 led to changes in
the expression levels of B. thetaiotaomicron genes during gut
colonization. However, colonization of the BT2391 mutant
promoted DSS colitis in low-fiber diet-fed mice. These results
indicate that BT2391 contributes to a sustainable symbiotic
relationship by maintaining a balance between mucosal
colonization and gut homeostasis.
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We evaluated the Cre-lox and CRISPR-Cas9 systems as markerrecycling
tools in Saccharomyces cerevisiae recombinants containing
multiple-integrated expression cassettes. As an initial
trial, we constructed rDNA-nontranscribed spacer- or Ty4-
based multiple integration vectors containing the URA3 marker
flanked by the loxP sequence. Integrants harboring multiple
copies of tHMG1 and NNV-CP expression cassettes were obtained
and subsequently transformed with the Cre plasmid.
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along with the URA3 marker hampered the use of Cre-lox as
a marker-recycling tool in multiple integrants. As an alternative,
we constructed a set of CRISPR-Cas9-gRNA vectors containing
gRNA targeted to auxotrophic marker genes. Transformation
of multiple integrants of tHMG1 and NNV-CP
cassettes by the Cas9-gRNA vector in the presence of the URA3
(stop) donor DNA fragments generated the Ura- transformants
retaining multiple copies of the expression cassettes.
CRISPR-Cas9-based inactivation led to the recycling of the
other markers, HIS3, LEU2, and TRP1, without loss of expression
cassettes in the recombinants containing multiple
copies of tHMG1, NNV-CP, and SfBGL1 cassettes, respectively.
Reuse of the same selection marker in marker-inactivated
S. cerevisiae was validated by multiple integrations of the
TrEGL2 cassette into the S. cerevisiae strain expressing SfBGL1.
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selection marker genes using the CRISPR-Cas9 system with
donor DNA fragments is an efficient strategy for markerrecycling
in multiple integrants. In particular, the continual
reuse of auxotrophic markers would facilitate the construction
of a yeast cell factory containing multiple copies of expression
cassettes without antibiotic resistance genes.
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poses major threats to public health worldwide. With increasing
concerns about the limitations of current disinfectant treatments,
phage-derived depolymerases may be used as promising
biocontrol agents. Therefore, in this study, the characterization,
purification, and application of a novel phage depolymerase,
Dpo10, specifically targeting the lipopolysaccharides
of E. coli O157, was performed. Dpo10, with a molecular
mass of 98 kDa, was predicted to possess pectate lyase
activity via genome analysis and considered to act as a receptor-
binding protein of the phage. We confirmed that the
purified Dpo10 showed O-polysaccharide degrading activity
only for the E. coli O157 strains by observing its opaque halo.
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range of temperature conditions under 55°C and mild basic
pH. Notably, Dpo10 did not inhibit bacterial growth but significantly
increased the complement-mediated serum lysis
of E. coli O157 by degrading its O-polysaccharides. Moreover,
Dpo10 inhibited the biofilm formation against E. coli O157
on abiotic polystyrene by 8-fold and stainless steel by 2.56 log
CFU/coupon. This inhibition was visually confirmed via fieldemission
scanning electron microscopy. Therefore, the novel
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The COVID-19 pandemic has caused unprecedented health,
social, and economic crises worldwide. However, to date, there
is an only a limited effective treatment for this disease. Human
placenta hydrolysate (hPH) has previously been shown to be
safe and to improve the health condition in patients with hyperferritinemia
and COVID-19. In this study, we aimed to
determine the antiviral effects of hPH against SARS-CoV-2
in vitro and in vivo models and compared with Remdesivir,
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whether hPH inhibited SARS-CoV-2 replication, we determined
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by infection with a SARS-CoV-2 at an MOI of 0.01. Further,
groups of ferrets infected with 105.8 TCID50/ml of SARS-CoV-2
and treated with hPH at 2, 4, 6 dpi, and compared their clinical
manifestation and virus titers in respiratory tracts with
PBS control-treated group. The mRNA expression of immunerelated
cytokines was determined by qRT-PCR. hPH treatment
attenuated virus replication in a dose-dependent manner in
vitro. In a ferret infection study, treatment with hPH resulted
in minimal bodyweight loss and attenuated virus replication
in the nasal wash, turbinates, and lungs of infected ferrets.
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remarkably upregulated the gene expression of type I
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ferrets. Our data collectively suggest that hPH has antiviral
efficacy against SARS-CoV-2 and might be a promising
therapeutic agent for the treatment of SARS-CoV-2 infection.
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Arthrobotrys oligospora is a model species of nematophagous
fungi and has great potential for the biological control of nematode
diseases. Lectin is a protein that binds to carbohydrates
and their complexes with high specificity, which mediates recognition
events in various physiological and pathological
processes. This study aimed to investigate the role of the
Jacalin-related lectin (JRL) gene, AOL_s00083g511, in A. oligospora
development. Through a homology recombination
approach, we obtained the AOL_s00083g511 knockout mutant
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,
3D-trap formation, and the ability of A. oligospora to
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.
The growth rates of both strains on high salt or surfactant media
were similar; however, in the strong oxidation medium,
the growth rate of the Δg511 mutant was significantly lower
than that of the WT strain, indicating that AOL_s00083g511
might play a role in oxidative stress resistance. These findings
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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The fucose-specific lectin gene AOL_s00054g276 affects trap formation and nematocidal activity of the nematophagous fungus Arthrobotrys oligospora
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Phospholipase C (AoPLC2) regulates mycelial development, trap morphogenesis, and pathogenicity of the nematode-trapping fungus Arthrobotrys oligospora Meihua Xie, Ni Ma, Na Bai, Meichen Zhu, Ke-Qin Zhang, Jinkui Yang Journal of Applied Microbiology.2022; 132(3): 2144. CrossRef
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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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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One of the advantages for initial survival of inhaled fungal
spores in the respiratory tract is the ability for iron acquisition
via hemolytic factor-production. To examine the ability
of indoor Aspergillus and Penicillium affecting hemolysis,
the secreted factors during the growth of thirteen strains from
eight species were characterized in vitro for their hemolytic
activity (HA) and CAMP-like reaction. The hemolytic index
of HA on human blood agar of Aspergillus micronesiensis,
Aspergillus wentii, Aspergillus westerdijkiae, Penicillium citrinum,
Penicillium copticola, Penicillium paxilli, Penicillium
steckii, and Penicillium sumatrense were 1.72 ± 0.34, 1.61 ±
0.41, 1.69 ± 0.16, 1.58 ± 0.46, 3.10 ± 0.51, 1.22 ± 0.19, 2.55 ±
0.22, and 1.90 ± 0.14, respectively. The secreted factors of
an Aspergillus wentii showed high HA when grown in undernourished
broth at 25°C at an exponential phase and were
heat sensitive. Its secreted proteins have an estimated relative
molecular weight over 50 kDa. Whereas, the factors of
Penicillium steckii were secreted in a similar condition at a
late exponential phase but showed low HA and heat tolerance.
In a CAMP-like test with sheep blood, the synergistic hemolytic
reactions between most tested mold strains and Staphylococcus
aureus were identified. Moreover, the enhancement
of α-hemolysis of Staphylococcus aureus could occur through
the interaction of Staphylococcus aureus-sphingomyelinase
and CAMP-like factors secreted from Aspergillus micronesiensis.
Further studies on the characterization of purified hemolytic-
and CAMP-like-factors secreted from Aspergillus
wentii and Aspergillus micronesiensis may lead to more understanding
of their involvement of hemolysis
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target genes that govern fungal differentiation and cellular
metabolism. In this study, we characterize one of the
VosA/VelB-inhibited developmental genes called vidA, which
is predicted to encode a 581-amino acid protein with a C2H2
zinc finger domain at the C-terminus. Levels of vidA mRNA
are high during the early and middle phases of asexual development
and decrease during the late phase of asexual development
and asexual spore (conidium) formation. Deletion
of either vosA or velB results in increased vidA mRNA accumulation
in conidia, suggesting that vidA transcript accumulation
in conidia is repressed by VosA and VelB. Phenotypic
analysis demonstrated that deletion of vidA causes decreased
colony growth, reduced production of asexual spores,
and abnormal formation of sexual fruiting bodies. In addition,
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that VidA is a putative transcription factor that plays a
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SMF 134, which was isolated from meju (Korean soybean
fermented brick), was analyzed at the genomic level to evaluate
its potential as a starter for soybean fermentation. The
genome size was 40.1 Mbp, which was expected to be composed
of eight chromosomes with 13,748 ORFs. Strain SMF
134 had a total of 151 protease genes, among which two more
leucine aminopeptidase (lap) genes were found in addition to
the previously known lap1, and three γ-glutamyltranspeptidase
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fermentation. In addition, this first complete genome of
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respectively.
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