Intergenic recombination in feline calicivirus associated with a hemorrhagic-like disease in the Republic of Korea.

Feline calicivirus (FCV) is a common cause of upper respiratory tract disease in cats. In this study, the complete genome sequence of FCV 14Q315, which was detected from a dead domestic cat with a hemorrhagic-like disease, was analyzed to identify the genetic characteristics. The FCV 14Q315 genome was 7,684 bp. Phylogenetic analyses based on the ORF1, ORF2, and ORF3 sequences indicated that FCV 14Q315 is more closely related to FCV 15D022 than to other FCV strains. ORF1 of FCV 14Q315 shared high sequence similarity with ORF1 of FCVs 15D022 and UTCVM-H1. We further evaluated genetic recombination in ORF1 of FCV 14Q315 and detected intergenic recombination between p30 and the ORF1/ORF2 junction with high significance. Particularly, the non-recombination region in ORF1 of FCV 14Q315 showed high sequence similarity with FCVs GX2019, CH-JL2, and 15D022. The recombination region in ORF1 of FCV 14Q315 showed the highest similarity with FCV UTCVM-H1, which is associated with a hemorrhagic-like disease. The results suggest that the UTCVM-H1-like FCV was introduced into the Republic of Korea and presumably recombined with Korean FCVs by occasional mixed infections. In addition, the Korean FCV strains were located in several phylogenetic clusters with marked genetic diversity in the ORF2 region. These results imply that Korean FCVs possess high genetic diversity owing to mutations and recombination. Furthermore, it is possible that certain FCVs caused cyclical infections in the Korean cat population based on a phylogenetic analysis of FCVs isolated at different time points. Keywords: calicivirus; virulent systemic feline calicivirus; recombination; hemorrhagic-like disease.

Genetic diversity and phylogenetic analysis of newly discovered bat astroviruses in Korea.

Bats have been identified as a natural reservoir for several potentially zoonotic viruses. Recently, astroviruses have been reported in bats in many countries, but not Korea. We collected 363 bat samples from thirteen species at twenty-nine sites in Korea across 2016 and tested them for astrovirus. The detection of the RNA-dependent RNA polymerase (RdRp) gene in bat astroviruses was confirmed in thirty-four bats across four bat species in Korea: twenty-five from Miniopterus fuliginosusi, one from Myotis macrodactylus, four from M. petax, and four from Rhinolophus ferrumequinum. The highest detection rates for astrovirus were found in Sunchang (61.5%, 8/13 bats), and in the samples collected in April (63.2%, 12/19 bats). The amino acid identity of astroviral sequences identified from bat samples was ≥ 46.6%. More specifically, the amino acid identity within multiple clones from individual bats was ≥ 50.8%. Additionally, the phylogenetic topology between astroviruses from different bat families showed a close relationship. Furthermore, phylogenetic analysis of the partial ORF2 sequence of bat astroviruses was found to have a maximum similarity of 73.3-74.8% with available bat astrovirus sequences. These results indicate potential multiple-infection by several bat astrovirus species in individual bats, or hyperpolymorphism in the astrovirus strains, as well as the transmission of astroviruses across bat families; furthermore, our phylogenetic analysis of the partial ORF2 implied that a novel astrovirus may exist. However, the wide diversity of astroviral sequences appeared to have no significant correlation with bat species or the spatiotemporal distribution of Korean bat astroviruses.

Molecular characterization of the spike and ORF3 genes of porcine epidemic diarrhea virus in the Philippines.

Porcine epidemic diarrhea virus (PEDV) is a highly contagious enteric pathogen of swine. In the present study, we analyzed the spike genes and ORF3 genes of seven PEDV strains detected in Philippine pigs in June 2014. There are four major epitope regions in the spike glycoprotein: a CO-26K equivalent (COE) domain, SS2 and SS6 epitopes, and an epitope region recognized by the 2C10 monoclonal antibody. Analysis of Philippine strains revealed amino acid substitutions in the SS6 epitope region (LQDGQVKI to SQSGQVKI) of the S1 domain. Substitutions were also detected in the 2C10 epitope region (GPRLQPY to GPRFQPY) in the cytoplasmic domain. Phylogenetic analysis of the complete spike gene sequences from the seven strains revealed that they clustered within the G2 group but were distantly related to the North American and INDELs clusters. Interestingly, these strains were close to Vietnamese PEDVs on the ORF3 genetic tree and showed high (97.0-97.6 %) sequence identity to ORF3 genes at the nucleotide level.

Molecular evolution of kobuviruses in cats.

Aichi virus, a causative agent of human gastroenteritis, is one of a number of animal viruses belonging to the genus Kobuvirus within the family Picornaviridae. The kobuvirus genome encodes several structural and nonstructural proteins; the capsid proteins encoded by the VP1 gene are key immunogenic factors. Here, we used the VP1 region to determine substitution rates and the time to the most recent common ancestor (TMRCA) by comparing feline kobuvirus (FKoVs) sequences with kobuvirus sequences isolated from members of other species. The substitution rate for FKoVs was 1.29 × 10(-2 )substitutions/site/year (s/s/y) and the TMRCA was 5.3 years.

A novel strain of porcine epidemic diarrhea virus in Vietnamese pigs.

Porcine epidemic diarrhea virus (PEDV) causes severe diarrhea and dehydration in suckling pigs and has caused high rates of death among piglets and substantial economic loss in Vietnam since 2009. To investigate the genotypes of prevailing PEDVs, intestinal and fecal samples from piglets from central and northern Vietnam were collected and analyzed. Phylogenetic analysis of the nucleotide sequences of complete spike genes of PEDVs from Vietnam resulted in the identification of two divergent groups. PEDVs (HUA-PED45 and HUA-PED47) belonged to the G2b group, along with Chinese, US, and Korean strains occurring at the end of 2010, in May 2013 and in November 2013, respectively. Six strains from the Quang Tri region were assigned to the G1b group, along with Chinese and US strains. The Vietnamese PEDVs detected in infected piglets had a nationwide distribution and belonged to the G2b and G1b genotypes.

Serological prevalence of viral agents that induce reproductive failure in South Korean wild boar.

BACKGROUND: Viral agents associated with reproductive failure such as Aujeszky's disease virus (ADV), encephalomyocarditis virus (EMCV), and porcine parvovirus (PPV) have also been identified in European wild boar. To screen for the presence of antibodies against ADV, EMCV, and PPV from wild boar (Sus scrofa) in South Korea, 481 serum samples were collected from wild boar hunted between December 2010 and May 2011. RESULTS: Of the 481 serum samples tested, 47 (9.8%) and 37 (7.7%) were seropositive for ADV and EMCV antibodies, respectively, based on a neutralization test (VNT), and 142 (29.5%) were seropositive for PPV antibodies based on a hemagglutination inhibition (HI) test. CONCLUSIONS: This was the first survey to identify the seroprevalence of the three major viruses associated with reproductive failure in the wild boar population of South Korea. Wild boar may act as a reservoir for many viruses that cause infectious diseases in domestic pigs. Thus, strict prevention and control measures, such as continuous wildlife disease surveillance and strategic methods of downsizing the population density, should be implemented to prevent disease transmission from wild boar to domestic pigs.

Effects of diets for three growing stages by rumen inocula donors on in vitro rumen fermentation and microbiome.

Hanwoo and Jeju Black cattle (Jeju Black) are native breeds of Korean cattle. Jeju Black cattle are recognized as natural monuments and are known to exhibit slower growth rates compared to Hanwoo. While several studies have analyzed the genetic characteristics of these cattle, there has been limited research on the differences in their microbiome. In this study, rumen fluid was obtained from three Hanwoo steers and three Jeju Black steers, and three different diets (total mixed rations [TMRs] for growing, early fattening, and late fattening periods) were used as substrates for in vitro fermentation. The in vitro incubation was conducted for 3 h and 24 h following a 2 × 3 factorial arrangement. After both incubation periods, fermentation characteristics were analyzed, and ruminal microbiome analysis was performed using 16S rRNA gene sequencing, employing both QIIME2 and PICRUSt2. The results revealed significant differences in the ruminal microbiota due to the inoculum effect. At the phylum level, Patescibacteria and Synergistota were found to be enriched in the Jeju Black inoculum-treated group. Additionally, using different inocula also affected the relative abundance of major taxa, including Ruminococcus, Pseudoramibacter, Ruminococcaceae CAG-352, and the [Eubacterium] ruminantium group. These microbial differences induced by the inoculum may have originated from varying levels of domestication between the two subspecies of donor animals, which mainly influenced the fermentation and microbiome features in the early incubation stages, although this was only partially offset afterward. Furthermore, predicted commission numbers of microbial enzymes, some of which are involved in the biosynthesis of secondary metabolites, fatty acids, and alpha amylase, differed based on the inoculum effect. However, these differences may account for only a small proportion of the overall metabolic pathway. Conversely, diets were found to affect protein biosynthesis and its related metabolism, which showed differential abundance in the growing diet and were potentially linked to the growth-promoting effects in beef cattle during the growing period. In conclusion, this study demonstrated that using different inocula significantly affected in vitro fermentation characteristics and microbiome features, mainly in the early stages of incubation, with some effects persisting up to 24 h of incubation.

Intranasal immunization with the recombinant measles virus encoding the spike protein of SARS-CoV-2 confers protective immunity against COVID-19 in hamsters.

BACKGROUND: As the nasal mucosa is the initial site of infection for COVID-19, intranasal vaccines are more favorable than conventional vaccines. In recent clinical studies, intranasal immunization has been shown to generate higher neutralizing antibodies; however, there is a lack of evidence on sterilizing immunity in the upper airway. Previously, we developed a recombinant measles virus encoding the spike protein of SARS-CoV-2 (rMeV-S), eliciting humoral and cellular immune responses against SARS-CoV-2. OBJECTIVES: In this study, we aim to provide an experiment on nasal vaccines focusing on a measles virus platform as well as injection routes. STUDY DESIGN: Recombinant measles viruses expressing rMeV-S were prepared, and 5 × 105 PFUs of rMeV-S were administered to Syrian golden hamsters via intramuscular or intranasal injection. Subsequently, the hamsters were challenged with inoculations of 1 × 105 PFUs of SARS-CoV-2 and euthanized 4 days post-infection. Neutralizing antibodies and RBD-specific IgG in the serum and RBD-specific IgA in the bronchoalveolar lavage fluid (BALF) were measured, and SARS-CoV-2 clearance capacity was determined via quantitative reverse-transcription PCR (qRT-PCR) analysis and viral titer measurement in the upper respiratory tract and lungs. Immunohistochemistry and histopathological examinations of lung samples from experimental hamsters were conducted. RESULTS: The intranasal immunization of rMeV-S elicits protective immune responses and alleviates virus-induced pathophysiology, such as body weight reduction and lung weight increase in hamsters. Furthermore, lung immunohistochemistry demonstrated that intranasal rMeV-S immunization induces effective SARS-CoV-2 clearance that correlates with viral RNA content, as determined by qRT-PCR, in the lung and nasal wash samples, SARS-CoV-2 viral titers in lung, nasal wash, BALF samples, serum RBD-specific IgG concentration, and RBD-specific IgA concentration in the BALF. CONCLUSION: An intranasal vaccine based on the measles virus platform is a promising strategy owing to the typical route of infection of the virus, the ease of administration of the vaccine, and the strong immune response it elicits.

High-resolution field effect sensing of ferroelectric charges.

Nanoscale manipulation of surface charges and their imaging are essential for understanding local electronic behaviors of polar materials and advanced electronic devices. Electrostatic force microscopy and Kelvin probe force microscopy have been extensively used to probe and image local surface charges responsible for electrodynamics and transport phenomena. However, they rely on the weak electric force modulation of cantilever that limits both spatial and temporal resolutions. Here we present a field effect transistor embedded probe that can directly image surface charges on a length scale of 25 nm and a time scale of less than 125 µs. On the basis of the calculation of net surface charges in a 25 nm diameter ferroelectric domain, we could estimate the charge density resolution to be as low as 0.08 µC/cm(2), which is equivalent to 1/20 electron per nanometer square at room temperature.

Image Shadow Detection and Removal Based on Region Matching of Intelligent Computing.

Shadow detection and removal play an important role in the field of computer vision and pattern recognition. Shadow will cause some loss and interference to the information of moving objects, resulting in the performance degradation of subsequent computer vision tasks such as moving object detection or image segmentation. In this paper, each image is regarded as a small sample, and then a method based on material matching of intelligent computing between image regions is proposed to detect and remove image shadows. In shadow detection, the proposed method can be directly used for detection without training and ensures the consistency of similar regions to a certain extent. In shadow removal, the proposed method can minimize the influence of shadow removal operation on other features in the shadow region. The experiments on the benchmark dataset demonstrate that the proposed approach achieves a promising performance, and its improvement is more than 6% in comparison with several advanced shadow detection methods.

Foreground Detection Based on Superpixel and Semantic Segmentation.

Foreground detection is an essential step in computer vision and video processing. Accurate foreground object extraction is crucial for subsequent high-level tasks such as target recognition and tracking. Although many foreground detection algorithms have been proposed, foreground detection in complex scenes is still a challenging problem. This paper presents a foreground detection algorithm based on superpixel and semantic segmentation. It first uses multiscale superpixel segmentation to obtain the initial foreground mask. At the same time, a semantic segmentation network is applied to separate potential foreground objects, and then use the defined rules to combine the results of superpixel and semantic segmentation to get the final foreground object. Finally, the background model is updated with the refined foreground result. Experiments on the CDNet2014 dataset demonstrate the effectiveness of the proposed algorithm, which can accurately segment foreground objects in complex scenes.

Complete genome analysis of the African swine fever virus isolated from a wild boar responsible for the first viral outbreak in Korea, 2019.

African swine fever (ASF), a highly contagious and severe hemorrhagic viral disease in swine, is emerging as a major threat not only in Korea but also worldwide. The first confirmed case of ASF in Korea was reported in 2019. Despite the occurrence of ASF in Korea, only a few studies have genetically characterized the causative ASF virus (ASFV). In this study, we aimed to genetically characterize the ASFV responsible for the 2019 outbreak in Korea. The genome of the ASFV isolated during the first outbreak in Korea was analyzed. The Korea/YC1/2019 strain has 188,950 base pairs, with a GC content of 38.4%. The complete genome sequence was compared with other ASFV genomes annotated in the NCBI database. The Korea/YC1/2019 strain shared the highest similarity with Georgia 2007, Belgium 2018/1, and ASFV-wbBS01 strains. This study expands our knowledge of the genetic diversity of ASFV, providing valuable information for epidemiology, diagnostics, therapies, and vaccine development.

Deletion of the bis gene results in a marked increase in the production of corticosterone that is associated with thymic atrophy in mice.

Bis (Bag3) is known to be involved in cell survival, migration, the regulating of chaperones, and protein quality control. We reported recently on the production of bis gene-deleted mice, which show early lethality within 3 wk after birth with a phenotype showing severe malnutrition and shrinkage of the thymus. In this report, we provide evidence to show that an intrinsic problem of adrenal gland is the the primary cause for the severe atrophy of the thymus in bis(-/-) mice. The bis(-/-) mice show significantly higher levels of corticosterone, but CRH and ACTH levels were considerably lower than those of wild littermates. The transcription of steroidogenic enzymes was increased in the adrenal glands of bis(-/-) mice, accompanied by an increase in the thickness of the zona reticularis. An analysis of thymus tissue from bis(-/-) mice revealed that the severe atrophy of the thymus is due to the specific loss of immature double-positive (CD4(+)CD8(+)) cortical thymocytes by apoptosis, as evidenced by immunohistochemical examination and flow cytometric analysis, which were restored by injection of an inhibitor of glucocorticoid synthesis. In vitro cultures of thymocytes with increasing doses of dexamethasone exhibited a similar degree of apoptosis between wild and bis(-/-) thymocytes. The corticosterone levels from fasted wild littermates were one-half those of bis(-/-) mice, although serum glucose levels were similar. Thus, the deletion of the bis gene resulted in the intrinsic defect in the adrenal gland, leading to a marked increase in glucocorticoid levels, probably upon starvation stress, which accounts for the massive apoptosis of the thymus.

Nanoscale bit formation in highly (111)-oriented ferroelectric thin films deposited on glass substrates for high-density storage media.

PbTiO(3) (PTO) ferroelectric films on Pt(111) bottom electrode layers covering Ta/glass were prepared using pulsed laser deposition. X-ray diffraction patterns revealed that the PTO films were preferentially (111)-oriented. The films were highly crystalline and had a smooth surface with root mean square (RMS) roughness of 1.5 nm. Ferroelectric properties of the PTO films were characterized using piezoresponse force microscopy (PFM). PFM techniques achieved ferroelectric polarization bits with a minimum width of 22 nm, which corresponds to a potential recording density of 1.3 Tbit/in(2) in ferroelectric storage devices.

Luminance efficiency enhancement in green organic light-emitting devices fabricated utilizing a cesium fluoride/fullerene heterostructure electron injection layer.

Enhancement mechanisms of the luminance efficiency in green organic light-emitting devices (OLEDs) fabricated utilizing a cesium fluoride (CsF)/fullerene (C60) heterostructure acting as an electron injection layer (EIL) were investigated. The luminance efficiencies as functions of the current density showed that the luminance efficiency in the green OLEDs fabricated utilizing a CsF/C60 heterostructure acting as an EIL was higher than that in the green OLEDs fabricated utilizing a CsF, a Liq, or a C60 single EIL. The interfacial dipole existing at the CsF layer decreased the electron injection barrier, and the stability of the OLEDs with a CsF EIL was enhanced due to the lower diffusion rate of Cs atoms in comparison with Li atoms. The enhancement of the luminance efficiency of the OLEDs with a heterostructure EIL was attributed to the increase in the electron injection. These results can help improve understanding of the enhancement mechanisms of the luminance efficiency in green OLEDs utilizing a CsF/C60 heterostructure acting as an EIL.

Wild boar harbouring African swine fever virus in the demilitarized zone in South Korea, 2019.

The African swine fever virus (ASFV) was first detected in wild boar in the Demilitarized Zone, a bordered area between South and North Korea, on 2 October 2019. Phylogenetic analyses of ASFV genes encoding p72 and CD2v indicated that the causative strain belongs to genotype II and serogroup 8, respectively, and contained additional tandem repeat sequences between the I73R and the I329L protein genes.

Fabrication of nanowire channels with unidirectional alignment and controlled length by a simple, gas-blowing-assisted, selective-transfer-printing technique.

A printing-based lithographic technique for the patterning of V(2)O(5) nanowire channels with unidirectional orientation and controlled length is introduced. The simple, directional blowing of a patterned polymer stamp with N(2) gas, inked with randomly distributed V(2)O(5) nanowires, induces alignment of the nanowires perpendicular to the long axis of the line patterns. Subsequent stamping on the amine-terminated surface results in the selective transfer of the aligned nanowires with a controlled length corresponding to the width of the relief region of the polymer stamp. By employing such a gas-blowing-assisted, selective-transfer-printing technique, two kinds of device structures consisting of nanowire channels and two metal electrodes with top contact, whereby the nanowires were aligned either parallel (parallel device) or perpendicular (serial device) to the current flow in the conduction channel, are fabricated. The electrical properties demonstrate a noticeable difference between the two devices, with a large hysteresis in the parallel device but none in the serial device. Systematic analysis of the hysteresis and the electrical stability account for the observed hysteresis in terms of the proton diffusion in the water layer of the V(2)O(5) nanowires, induced by the application of an external bias voltage higher than a certain threshold voltage.

Genetic Characterization of Canine Parvovirus Type 2 Detected in Wild Raccoon Dogs ( Nyctereutes procyonoides) in the Republic of Korea.

Canine parvovirus (CPV) was detected in three of 136 samples from dead raccoon dogs ( Nyctereutes procyonoides) in the Republic of Korea (South Korea) during 2016-17. By sequence and phylogenetic analysis of the complete VP2 gene, the strain belonged to CPV-2 and would be distinct from the previous reported CPV-2a and CPV-2b strains from Korean domestic dogs ( Canis lupus familiaris). The results indicated that the CPV strains from raccoon dogs and domestic dogs might be not circulated between wild and domestic carnivores in Korea.

Minimizing an outbreak of avian botulism (Clostridium botulinum type C) in Incheon, South Korea.

An outbreak of botulism occurred over a two-month period beginning July 20, 2016. In all, 697 wild birds were found paralyzed or dead at the Namdong reservoir and 11 Gong-gu. Using a mouse bioassay, type C botulinum toxin was identified in the bird serum, liquid cultures of soil samples, and maggot extracts. To minimize further infection of wild birds, we opened the floodgates of the Namdong reservoir adjacent to the Yellow Sea; this decreased the water temperature and the nutrient load such as nitrogen and phosphorus. The outbreak stopped shortly after taking these actions. It is not known if these efforts decreased the number of dead and diseased wild birds. Our study demonstrates one potential approach to minimize future botulism outbreaks among wild birds and their habitats.

Antigenic characterization of classical swine fever virus YC11WB isolates from wild boar.

Classical swine fever (CSF), a highly contagious disease that affects domestic pigs and wild boar, has serious economic implications. The present study examined the virulence and transmission of CSF virus strain YC11WB (isolated from a wild boar in 2011) in breeding wild boar. Virulence of strain YC11WB in domestic pigs was also examined. Based on the severe clinical signs and high mortality observed among breeding wild boar, the pathogenicity of strain YC11WB resembled that of typical acute CSF. Surprisingly, in contrast to strain SW03 (isolated from breeding pigs in 2003), strain YC11WB showed both acute and strong virulence in breeding pigs. None of three specific monoclonal antibodies (7F2, 7F83, and 6F65) raised against the B/C domain of the SW03 E2 protein bound to the B/C domain of strain YC11WB due to amino acid mutations (720K→R and 723N→S) in the YC11WB E2 protein. Although strains YC11WB and SW03 belong to subgroup 2.1b, they had different mortality rates in breeding pigs. Thus, if breeding pigs have not developed protective immunity against CSF virus, they may be susceptible to strain YC11WB transmitted by wild boar, resulting in severe economic losses for the pig industry.