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result(s) for
"Suzuki, Tohru"
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Genomic Characterization and Phylogenetic Classification of Bovine Coronaviruses Through Whole Genome Sequence Analysis
by
Otake, Yoshihiro
,
Suzuki, Tohru
,
Hasebe, Ayako
in
bovine coronavirus
,
genotype classification
,
phylogenetic analysis
2020
Bovine coronavirus (BCoV) is zoonotically transmissible among species, since BCoV-like viruses have been detected in wild ruminants and humans. BCoV causing enteric and respiratory disease is widespread in cattle farms worldwide; however, limited information is available regarding the molecular characterization of BCoV because of its large genome size, despite its significant economic impact. This study aimed to better understand the genomic characterization and evolutionary dynamics of BCoV via comparative sequence and phylogenetic analyses through whole genome sequence analysis using 67 BCoV isolates collected throughout Japan from 2006 to 2017. On comparing the genomic sequences of the 67 BCoVs, genetic variations were detected in 5 of 10 open reading frames (ORFs) in the BCoV genome. Phylogenetic analysis using whole genomes from the 67 Japanese BCoV isolates in addition to those from 16 reference BCoV strains, revealed the existence of two major genotypes (classical and US wild ruminant genotypes). All Japanese BCoV isolates originated from the US wild ruminant genotype, and they tended to form the same clusters based on the year and farm of collection, not the disease type. Phylogenetic trees on hemagglutinin-esterase protein (HE), spike glycoprotein (S), nucleocapsid protein (N) genes and ORF1 revealed clusters similar to that on whole genome, suggesting that the evolution of BCoVs may be closely associated with variations in these genes. Furthermore, phylogenetic analysis of BCoV S genes including those of European and Asian BCoVs and human enteric coronavirus along with the Japanese BCoVs revealed that BCoVs differentiated into two major types (European and American types). Moreover, the European and American types were divided into eleven and three genotypes, respectively. Our analysis also demonstrated that BCoVs with different genotypes periodically emerged and predominantly circulated within the country. These findings provide useful information to elucidate the detailed molecular characterization of BCoVs, which have spread worldwide. Further genomic analyses of BCoV are essential to deepen the understanding of the evolution of this virus.
Journal Article
Multiplex RT-qPCR Application in Early Detection of Bovine Respiratory Disease in Healthy Calves
2023
Bovine respiratory diseases (BRD) are associated with various predisposing factors, such as physical and physiological stress factors, and bacterial and viral pathogens. These stressors and viruses suppress immune defenses, leading to bacterial growth in the upper respiratory tract and invasion of pathogens into the lower respiratory tract. Therefore, continuous monitoring of the causative pathogens would contribute to the early detection of BRD. Nasal swabs and sera from 63 clinically healthy calves were continuously collected from seven farms in Iwate prefecture from 2019 to 2021. We attempted to monitor dynamics of BRD-associated pathogens by multiplex real-time RT-PCR (RT-qPCR) using their nasal swab samples. In addition, we attempted to monitor fluctuation of antibody titers against each BRD-associated pathogen by virus neutralization test (VNT) using their sera. In contrast, nasal swabs from 89 calves infected with BRD were collected from 28 farms in Iwate prefecture from 2019 to 2021. We attempted to analyze their nasal swab samples by multiplex RT-qPCR aim to detect BRD-associated pathogens that are dominant in this region. As a result, our analyses using samples from clinically healthy calves showed that positive results by multiplex RT-qPCR were closely related to a significant increase of antibody titers by VNT in bovine coronavirus (BCoV), bovine torovirus (BToV), and bovine respiratory syncytial virus (BRSV). In addition, our data exhibited that BCoV, BToV, BRSV, bovine parainfluenza virus 3, and Mycoplasma bovis have been more frequently detected in calves infected with BRD compared to those detected in clinically healthy calves. Moreover, the data presented herein revealed co-infections by combination multiple viral pathogens with bacterial pathogens are closely involved in the onset of BRD. Taken together, our study demonstrates multiplex RT-qPCR which can simultaneously analyze multiple pathogens, including viruses and bacteria, and is useful for the early detection of BRD.
Journal Article
In-situ scanning transmission electron microscopy study of Al-amorphous SiO2 layer-SiC interface
by
Nakayama, Yoshiko
,
Nishimiya, Yuki
,
Suzuki, Tohru S.
in
Aluminum carbide
,
Aluminum oxide
,
bias
2023
Here, we present a comprehensive study on atomic-scale in-situ biasing/heating scanning transmission electron microscopy ((S)TEM) of Al-amorphous SiO
2
–SiC interface. The investigation includes electrical, chemical, and structural analysis of the interface at different temperatures (25–600 °C). The results show that at ~ 500 °C the electrical (three-orders of magnitude resistivity drop), chemical (dissolution of SiO
2
amorphous layer), and microstructural features (
e.g.
formation of Al
2
O
3
, Si and Al
4
C
3
) of the interface start to change. According to the results, amorphous SiO
2
dissolves in Al, leading to formation of α-Al
2
O
3
and Si within the Al. In contrast, elemental interdiffusion (Al
3+
⇄ Si
4+
) between Al and SiC occurs resulting in formation of Al
4
C
3
. From the results, we can infer that reaction mechanism between Al and crystalline SiC is different with that between Al and SiO
2
amorphous phase. It is believed that structural similarities between SiC and Al
4
C
3
play an important role in paving the way for elemental interdiffusion.
Journal Article
Bifidobacteria can protect from enteropathogenic infection through production of acetate
by
Itoh, Kikuji
,
Toh, Hidehiro
,
Kikuchi, Jun
in
631/326/41/1969
,
631/326/41/2482
,
692/699/255/1318
2011
Protective effect of probiotic bacteria
Bifidobacteria, sometimes used in yoghurts and other food products as 'probiotics', are natural inhabitants of the human gut and are known to protect us from infection. A possible mechanism for at least part of that protective action has now been found in the form of acetate. Oral application of a subtype of
Bifidobacterium longum
in mice seems to protect them from potentially fatal
Escherichia coli
O157:H7 by promoting host-defence mechanisms in epithelial cells.
Bifidobacteria are natural inhabitants of the human gut and are known to provide protection from infection. It is now shown that certain features of bifidobacterial metabolism that ultimately lead to the production of acetate are involved in this effect.
The human gut is colonized with a wide variety of microorganisms, including species, such as those belonging to the bacterial genus
Bifidobacterium
, that have beneficial effects on human physiology and pathology
1
,
2
,
3
. Among the most distinctive benefits of bifidobacteria are modulation of host defence responses and protection against infectious diseases
4
,
5
,
6
. Nevertheless, the molecular mechanisms underlying these effects have barely been elucidated. To investigate these mechanisms, we used mice associated with certain bifidobacterial strains and a simplified model of lethal infection with enterohaemorrhagic
Escherichia coli
O157:H7, together with an integrated ‘omics’ approach. Here we show that genes encoding an ATP-binding-cassette-type carbohydrate transporter present in certain bifidobacteria contribute to protecting mice against death induced by
E. coli
O157:H7. We found that this effect can be attributed, at least in part, to increased production of acetate and that translocation of the
E. coli
O157:H7 Shiga toxin from the gut lumen to the blood was inhibited. We propose that acetate produced by protective bifidobacteria improves intestinal defence mediated by epithelial cells and thereby protects the host against lethal infection.
Journal Article
Clinical Analysis for Long-Term Sporadic Bovine Viral Diarrhea Transmitted by Calves with an Acute Infection of Bovine Viral Diarrhea Virus 2
2021
Bovine viral diarrhea virus (BVDV) is a viral pathogen associated with serious problems in the cattle industry. Cattle persistently infected (PI) with BVDV are mild or asymptomatic; however, they become a source of BVDV transmission to other cattle. Hence, it is important to rapidly identify and remove the PI animals from cattle herds. Whereas cattle acutely infected (AI) with BVDV have various symptoms, yet they generally recover within 3 weeks. However, there is a paucity of information concerning clinical characteristics of AI cattle. Further accumulation of information would be required to accurately diagnose AI cattle with BVDV. Here, we attempted to obtain valuable information via various analyses using a case report of BVD outbreak that occurred for approximately four months in Iwate Prefecture in 2017. Using eight calves and multiple tests (real-time RT-PCR, virus isolation, enzyme-linked immunosorbent assay, and virus neutralization assay) over 6 weeks, we diagnosed the continuous BVD outbreak as an acute infection and not a persistent one. Additionally, we revealed that the sporadic case was caused by low pathogenic BVDV2 via BVDV genotyping and phylogenetic analysis. The data suggest that BVDV2 AI animals might also be a source of transmission to susceptible calves; hence, it might persist for a long period owing to multiple AI animals. These findings provide useful information to diagnose AI and PI cattle with BVDV in the field.
Journal Article
Characterization of Transparent Fluorapatite Ceramics Fabricated by Spark Plasma Sintering
by
Suzuki, Tohru S.
,
Morita, Koji
,
Kim, Byung-Nam
in
Apatite
,
Biocompatibility
,
Biological products
2022
Highly optically transparent polycrystalline fluorapatite ceramics with hexagonal crystal structures were fabricated via a liquid-phase synthesis of fluorapatite powder, followed by spark plasma sintering (SPS). The effect of sintering temperature, as observed using a thermopile, on the optical transmittance and microstructure of the ceramics was investigated in order to determine suitable sintering conditions. As a result, high optical transmittance was obtained in the SPS temperature range of 950–1100 °C. The highest optical transmittance was obtained for the ceramic sample sintered at 1000 °C, and its average grain size was evaluated at only 134 nm. The grain size dramatically increased with temperature, and the ceramics became translucent at SPS temperatures above 1200 °C. The mechanical and thermal properties of the ceramics were measured to evaluate the thermal shock parameter, which was found to be comparable to or slightly smaller than that of single-crystal fluorapatite. This transparent polycrystalline fluorapatite ceramic material should prove useful in a wide range of applications, for example as a biomaterial or optical/laser material, in the future. Furthermore, the knowledge obtained in this study should help to promote the application of this ceramic material.
Journal Article
S1 Subunit of Spike Protein from a Current Highly Virulent Porcine Epidemic Diarrhea Virus Is an Important Determinant of Virulence in Piglets
by
Ohashi, Seiichi
,
Kamitani, Wataru
,
Suzuki, Tohru
in
Amino acids
,
Artificial chromosomes
,
Cloning
2018
Base on the sequence of S genes, which encode spike proteins, we previously identified three different types (North American, S INDEL, and S large-DEL types) of porcine epidemic diarrhea virus (PEDV) that have re-emerged in Japan since 2013. Based on experimental infections with the North American and S large-DEL types, we also hypothesized that PEDV virulence may be linked to the S1 subunit of the S protein. To test this hypothesis, we have now assayed in gnotobiotic piglets various recombinant PEDVs generated by reverse genetics. Piglets inoculated with CV777 maintained in National Institute of Animal Health, along with piglets infected with a recombinant form of the same virus, developed subclinical to mild diarrhea. In contrast, severe watery diarrhea, dehydration, weight loss, astasia, and high mortality were observed in piglets inoculated with recombinant strains in which the S gene was partially or fully replaced with corresponding sequences from the highly virulent Japanese PEDV isolate OKN-1/JPN/2013. Indeed, symptoms resembled those in piglets inoculated with the OKN-1/JPN/2013, and were especially pronounced in younger piglets. Collectively, the data demonstrate that the S1 subunit of the S protein is an important determinant of PEDV virulence, and advance development of new vaccine candidate.
Journal Article
An Importance of Long-Term Clinical Analysis to Accurately Diagnose Calves Persistently and Acutely Infected by Bovine Viral Diarrhea Virus 2
by
Suzuki, Tohru
,
Yaegashi, Gakuji
,
Fukunari, Kazuhiro
in
5' Untranslated Regions
,
Acute Disease
,
acutely infected
2021
Bovine viral diarrhea virus (BVDV) infection results in a wide variety of clinical manifestations and is a pathogen that is able to cause huge economic losses in the cattle industry worldwide. It is important to identify cattle that are persistently infected (PI) by BVDV within the herd as early as possible because PI animals are the main reservoir of the virus. In contrast, cattle who are acutely infected (AI) with BVDV show various clinical signs, but most cattle show either mild symptoms or are asymptomatic. In general, AI and PI animals can be distinguished by repeat testing within an interval of at least 21 days. However, we found a rare case of a BVDV2-infected AI animal with long-term viral presence, making it indistinguishable from PI through two tests within an interval of 21 days. As a result, we diagnosed one infected animal as AI after 35 days from the initial sample collection via multiple analyses. Our findings recommend performing an additional test using samples that have been collected after 14–21 days from the second sample collection in cases where it is difficult to accurately differentiate an AI diagnosis from a PI diagnosis after only two tests. Additionally, our analysis exhibits that monitoring the number of copies of viruses with similar genomes in the sera by means of quantitative real-time RT-PCR through several sample collections periods might be useful to distinguish AI from PI. Furthermore, our data suggest that the AI animals with a long-term viral presence who show test results similar to those of PI animals might be the result of a coincidental combination of various factors that are present in cattle fields. These findings provide useful information that can be used to improve the diagnosis of BVDV in the field.
Journal Article
Robust and orange-yellow-emitting Sr-rich polytypoid α-SiAlON (Sr 3 Si 24 Al 6 N 40 :Eu 2+ ) phosphor for white LEDs
by
Hirosaki, Naoto
,
Funahashi, Shiro
,
Suzuki, Tohru S.
in
Calcium
,
Chemical synthesis
,
Efficiency
2024
Nitrides and oxynitrides isostructural to α-Si
N
(
-α-SiAlON,
= Sr, Ca, Li) possess superb thermally stable photoluminescence (PL) properties, making them reliable phosphors for high-power solid-state lighting. However, the synthesis of phase-pure Sr-α-SiAlON still remains a great challenge and has only been reported for Sr below 1.35 at.% as the large size of Sr
ions tends to destabilize the α-SiAlON structure. Here, we succeeded to synthesize the single-phase powders of a unique 'Sr-rich' polytypoid α-SiAlON (Sr
Si
Al
N
:Eu
) phosphor with three distinctive Sr/Eu luminescence sites using a solid-state remixing-reannealing process. The Sr content of this polytypoid structure exceeds those of a few previously reported structures by over 200%. The phase purity, composition, structure, and PL properties of this phosphor were investigated. A single phase can be obtained by firing the stoichiometric mixtures of all-nitride precursors at 2050°C under a 0.92 MPa N
atmosphere. The Sr
Si
Al
N
:Eu
shows an intense orange-yellow emission, with the emission maximum of 590 nm and internal/external quantum efficiency of 66%/52% under 400 nm excitation. It also has a quite small thermal quenching, maintaining 93% emission intensity at 150°C. In comparison to Ca-α-SiAlON:Eu
, this Sr counterpart shows superior quantum efficiency and thermal stability, enabling it to be an interesting orange-yellow down-conversion luminescent material for white LEDs. The experimental confirmation of the existence of such 'Sr-rich' SiAlON systems, in a single-phase powder form, paves the way for the design and synthesis of novel 'Sr-rich' SiAlON-based phosphor powders with unparalleled properties.
Journal Article
Vickers indentation tests on olivine: size effects
by
Hiraga, Takehiko
,
Suzuki, Tohru S.
,
Koizumi, Sanae
in
Crystallography
,
Crystallography and Scattering Methods
,
Crystals
2020
We conducted Vickers indentation tests on Fe-free (Mg
2
SiO
4
) and Fe-bearing (Mg
1.8
Fe
0.2
SiO
4
) olivine single crystals and high-density polycrystalline material with average grain sizes ranging from 170 to 890 nm. The Vickers microhardness (
H
v
) of the Fe-free polycrystalline material with the finest grain size is ~ 17 GPa at a load of 0.1 N, while that of the Fe-bearing single crystal is ~ 8 GPa at the largest load applied. Overall,
H
v
decreases with increasing grain size, load (indentation depth), and the presence of Fe. For each grain size,
H
v
is well characterized by a power law of the form
H
v
/
H
v
0
∝
l
-
x
, where
H
v
0
is the depth-independent value of
H
v
,
l
represents either grain size or indentation depth, and
x
is 0.09. Despite the small exponent value for each size effect, the nonlinear interaction of the two size effects results in large variations of
H
v
in our samples. We show that our semi-empirically derived relationship as a function of grain size and indentation depth explains the
H
v
of both polycrystalline and single-crystal olivine at any indentation conditions. Indentation fracture toughness of the finest-grained material is 0.8
MPa
m
1
/
2
, which increases slightly to 1.1
MPa
m
1
/
2
with increasing grain size, while the toughness of the single crystals varies from 0.5 to 0.8
MPa
m
1
/
2
depending on the crystallographic orientation of the fracture planes.
Journal Article