Screening and characterization of a novel linear B-cell epitope on orf virus F1L protein.

Background: Orf, also known as contagious ecthyma (CE), is an acute, contagious zoonotic disease caused by the orf virus (ORFV). The F1L protein is a major immunodominant protein on the surface of ORFV and can induce the production of neutralizing antibodies. Methods: The prokaryotic expression system was used to produce the recombinant F1L protein of ORFV, which was subsequently purified and used to immunize mice. Positive hybridoma clones were screened using an indirect enzyme-linked immunosorbent assay (ELISA). The reactivity and specificity of the monoclonal antibody (mAb) were verified through Western blot and indirect immunofluorescence (IFA). The linear antigenic epitope specific to the mAb was identified through Western blot, using truncated F1L proteins expressed in eukaryotic cells. A multiple sequence alignment of the ORFV reference strains was performed to evaluate the degree of conservation of the identified epitope. Results: After three rounds of subcloning, a mAb named Ba-F1L was produced. Ba-F1L was found to react with both the exogenously expressed F1L protein and the native F1L protein from ORFV-infected cells, as confirmed by Western blot and IFA. The mAb recognized the core epitope 103CKSTCPKEM111, which is highly conserved among various ORFV strains, as shown by homologous sequence alignment. Conclusion: The mAb produced in the present study can be used as a diagnostic reagent for detecting ORFV and as a basic tool for exploring the mechanisms of orf pathogenesis. In addition, the identified linear epitope may be valuable for the development of epitope-based vaccines.

Computational Strategies in Drug Discovery: Leveraging Subtractive Genomic Analysis for Target Identification.

The utilization of In-silico subtractive genomic analysis has emerged as an important and essential method in modern drug discovery and development since it significantly improves the process of identifying and validating potential targets for discovering novel therapeutic compounds to treat severe infections caused by Antimicrobial-resistant (AMR) - pathogenic species. This review provides a complete overview of the methodology, advantages, disadvantages, and prospects, associated with subtractive genomic research in the context of drug discovery and development. The initial phase of analysis encompasses the retrieval of data, which serves as a foundation for the subsequent data mining process in Phase 1. After data mining, Phase 2 utilizes subtractive channels for the target's non-homology and essentiality analysis. Phase 3 of the study aims to provide a comprehensive understanding of prospective targets by their qualitative characterization. Further, Phase 4 of the study emphasizes on conducting structure-based analyses, which involves the determination, refinement, evaluation, and validation of three-dimensional structures of the target proteins, along with their active site prediction and selection of the novel therapeutic compounds against that active site on the obtained targets through virtual screening and docking studies by utilizing various databases and servers. The therapeutic compounds obtained can be then validated by in vitro and in vivo testing, thereby establishing a connection between the computational predictions and real applications.

Systematic enhancement of protein crystallization efficiency by bulk lysine-to-arginine (KR) substitution.

Structural genomics consortia established that protein crystallization is the primary obstacle to structure determination using x-ray crystallography. We previously demonstrated that crystallization propensity is systematically related to primary sequence, and we subsequently performed computational analyses showing that arginine is the most overrepresented amino acid in crystal-packing interfaces in the Protein Data Bank. Given the similar physicochemical characteristics of arginine and lysine, we hypothesized that multiple lysine-to-arginine (KR) substitutions should improve crystallization. To test this hypothesis, we developed software that ranks lysine sites in a target protein based on the redundancy-corrected KR substitution frequency in homologs. This software can be run interactively on the worldwide web at https://www.pxengineering.org/. We demonstrate that three unrelated single-domain proteins can tolerate 5-11 KR substitutions with at most minor destabilization, and, for two of these three proteins, the construct with the largest number of KR substitutions exhibits significantly enhanced crystallization propensity. This approach rapidly produced a 1.9 Å crystal structure of a human protein domain refractory to crystallization with its native sequence. Structures from Bulk KR-substituted domains show the engineered arginine residues frequently make hydrogen-bonds across crystal-packing interfaces. We thus demonstrate that Bulk KR substitution represents a rational and efficient method for probabilistic engineering of protein surface properties to improve crystallization.

First identification of canine adenovirus 1 in mink and bioinformatics analysis of its 100 K protein.

Introduction: Animal trade favors the spreading of emerging canine adenovirus 1 (CAdV-1) in mink. Because the 100K protein is not exposed to the viral surface at any stage, it can be used to differentiate the vaccine from wild virus infection. However, no related research has been conducted. This study aimed to find evidence of CAdV-1 in mink and predict the character of the 100K protein in the current circulating CAdV-1 strain of mink. Method: In this experiment, the identification of CAdV-1, the phylogenetic tree, homology, and bioinformatics analysis of 100K were conducted. Results: The results showed that the CAdV-1 was identified in the mink and that its Fiber was located in a separate branch. It was closely related to strains isolated from Norwegian Arctic fox and Red fox. 100K was located in a separate branch, which had the closest genetic relationship with skunks, porcupines, raccoons, and hedgehogs and a far genetic relationship with the strains in dogs. 100K protein is an unstable and hydrophobic protein. It had evidence of selective pressure and recombination, 1 glycosylation site, 48 phosphorylation sites, 60 dominant B cell epitopes, and 9 peptides of MHC-I and MHC-II. Its subcellular localization was mainly in the endoplasmic reticulum and mitochondria. The binding sites of 100K proteins were DBP proteins and 33K proteins. Discussion: The stains in the mink were different from fox. The exploration of its genomic characteristics will provide us with a deeper understanding of the prevention of canine adenovirus.

Analysis of NDM-1 and IMP-8 carbapenemase producing Raoultella planticola clinical isolates.

Object of our study was to analyze the carriage of resistance genes in carbapenem-resistant Raoultella planticola (CRRP) by whole genome sequencing (WGS). Three strains of CRRP (named WF0027, WF3597 and WF3648) were collected for clinical analysis and susceptibility of antimicrobial agents was determined. The WGS of three strains was done by Illumina platform and strain identification was performed by average nucleotide identity, and the antibiotic resistance genes carried by the three strains were detected by ABRicate software. Whole genome data of 46 CRRP strains were downloaded from the National Center for Biotechnology Information (NCBI) database, and the evolutionary tree was constructed by genomic single nucleotide polymorphism together with this study strains. Antimicrobial susceptibility testing revealed that WF3597 and WF3648 were susceptible to tigecycline and colistin, while exhibited resistance to 24 antimicrobial agents. WF0027 was resistant to 18 antimicrobial agents. A total of 25 resistance genes were identified using ABRicate software. WF0027 carried blaIMP-8, whereas WF3597 and WF3648 carried blaNDM-1 carbapenem resistance gene. As predicted by the PlasmidFinder, WF3597 and WF3648 carried one plasmid IncFII(p14)_1_p14, whereas WF0027 carried five plasmids. Evolutionary tree results show all strains are clustered into six groups, the strains WF3597 and WF3648 belonged to the same evolutionary group (E clade) and WF0027 belonged to the F clade. Three CRRP strains in our study carried carbapenem resistance genes (blaNDM-1 or blaIMP-8) and were resistant to multiple antimicrobial agents, posing a significant challenge for clinical treatment.

The Integration of Genome-Wide Association Study and Homology Analysis to Explore the Genomic Regions and Candidate Genes for Panicle-Related Traits in Foxtail Millet.

Panicle traits are important factors affecting yield, and their improvement has long been a critical goal in foxtail millet breeding. In order to understand the genetic basis of panicle formation, a large-scale genome-wide association study (GWAS) was performed in this study for six panicle-related traits based on 706,646 high-polymorphism SNP loci in 407 accessions. As a result, 87 quantitative trait loci (QTL) regions with a physical distance of less than 100 kb were detected to be associated with these traits in three environments. Among them, 27 core regions were stably detected in at least two environments. Based on rice-foxtail millet homologous comparison, expression, and haplotype analysis, 27 high-confidence candidate genes in the QTL regions, such as Si3g11200 (OsDER1), Si1g27910 (OsMADS6), Si7g27560 (GS5), etc., affected panicle-related traits by involving multiple plant growth regulator pathways, a photoperiod response, as well as panicle and grain development. Most of these genes showed multiple effects on different panicle-related traits, such as Si3g11200 affecting all six traits. In summary, this study clarified a strategy based on the integration of GWAS, a homologous comparison, and haplotype analysis to discover the genomic regions and candidate genes for important traits in foxtail millet. The detected QTL regions and candidate genes could be further used for gene clone and marker-assisted selection in foxtail millet breeding.

Mechanisms Underlying the Virulence Regulation of Vibrio alginolyticus ND-01 pstS and pstB with a Transcriptomic Analysis.

Vibrio alginolyticus is a common opportunistic pathogen of fish, shrimp, and shellfish, and many diseases it causes can result in severe economic losses in the aquaculture industry. Causing host disease was confirmed by several virulence factors of V. alginolyticus. To date, there have been no reports on the effect of the pstS gene on its virulence regulation of V. alginolyticus. The virulence mechanism of target genes regulating V. alginolyticus is worthy of further study. Previous studies found that Fructus schisandrae (30 mg/mL) inhibited the growth of V. alginolyticus ND-01 (OD600 = 0.5) for 4 h, while the expressions of pstS and pstB were significantly affected by F. schisandrae stress. So, we speculated that pstS and pstB might be the virulence genes of V. alginolyticus, which were stably silenced by RNAi to construct the silencing strains pstS-RNAi and pstB-RNAi, respectively. After the expression of pstS or pstB gene was inhibited, the adhesion capacity and biofilm formation of V. alginolyticus were significantly down-regulated. The chemotaxis and biofilm formation ability of pstS-RNAi was reduced by 33.33% and 68.13% compared with the wild-type strain, respectively. Sequence alignment and homology analysis showed that pstS was highly conserved, which suggested that pstS played a vital role in the secretion system of V. alginolyticus. The pstS-RNAi with the highest silencing efficiency was selected for transcriptome sequencing. The Differentially Expressed Genes (DEGs) and GO terms were mapped to the reference genome of V. alginolyticus, including 1055 up-regulated genes and 1134 down-regulated genes. The functions of the DEGs were analyzed by GO and categorized into different enriched functional groups, such as ribosome synthesis, organelles, biosynthesis, pathogenesis, and secretion. These DEGs were then mapped to the reference KEGG pathways of V. alginolyticus and enriched in commonalities in the metabolic, ribosomal, and bacterial secretion pathways. Therefore, pstS and pstB could regulate the bacterial virulence of V. alginolyticus by affecting its adhesion, biofilm formation ability, and motility. Understanding the relationship between the expressions of pstS and pstB with bacterial virulence could provide new perspectives to prevent bacterial diseases.

Homology Characteristics of EEG and EMG for Lower Limb Voluntary Movement Intention.

In the field of lower limb exoskeletons, besides its electromechanical system design and control, attention has been paid to realizing the linkage of exoskeleton robots to humans via electroencephalography (EEG) and electromyography (EMG). However, even the state of the art performance of lower limb voluntary movement intention decoding still faces many obstacles. In the following work, focusing on the perspective of the inner mechanism, a homology characteristic of EEG and EMG for lower limb voluntary movement intention was conducted. A mathematical model of EEG and EMG was built based on its mechanism, which consists of a neural mass model (NMM), neuromuscular junction model, EMG generation model, decoding model, and musculoskeletal biomechanical model. The mechanism analysis and simulation results demonstrated that EEG and EMG signals were both excited by the same movement intention with a response time difference. To assess the efficiency of the proposed model, a synchronous acquisition system for EEG and EMG was constructed to analyze the homology and response time difference from EEG and EMG signals in the limb movement intention. An effective method of wavelet coherence was used to analyze the internal correlation between EEG and EMG signals in the same limb movement intention. To further prove the effectiveness of the hypothesis in this paper, six subjects were involved in the experiments. The experimental results demonstrated that there was a strong EEG-EMG coherence at 1 Hz around movement onset, and the phase of EEG was leading the EMG. Both the simulation and experimental results revealed that EEG and EMG are homologous, and the response time of the EEG signals are earlier than EMG signals during the limb movement intention. This work can provide a theoretical basis for the feasibility of EEG-based pre-perception and fusion perception of EEG and EMG in human movement detection.

Application of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in the analysis of the pollution homology of clean room in workshop / 公共卫生与预防医学

Objective To investigate the source of microbial contamination in the clean room of the workshop. Methods Microbiological sampling was carried out from the air, environment and personnel of the workshop. The samples were cultivated, the microorganisms were detected by MALDI-TOF-MS, and homology analysis was performed with the microbial identification system of the instrument. Results A total of 14 species and 41 strains of bacteria were detected. Nine strains of Staphylococcus epidermidis were selected for homology analysis, and the Staphylococcus epidermidis from personnel gloves and headgear had 94% homology. There was 83% homology among the staphylococcus epidermidis derived from the sedimentation bacteria, the ground environment and personnel, which was higher than the 71% of the standard strain. Conclusion The homology analysis demonstrates that the pollution in the clean room of the workshop mainly comes from personnel, and secondly comes from the environment outside the workshop. Enterprises need to strengthen management to prevent the occurrence of microbial contamination. MALDI-TOF-MS can be used for rapid detection of complex environmental bacteria and for homology analysis.

Urinary Tract Infections Caused by Fluconazole-Resistant Trichosporon japonicum in 2 Kidney Transplant Patients and Analysis of Their Homology.

Trichosporon spp. are emerging opportunistic agents that cause systemic diseases and life-threatening disseminated disease in immunocompromised hosts. Trichosporon japonicum is a highly rare cause of invasive trichosporonosis. In this study, we describe 2 cases of urinary tract infection caused by Trichosporon japonicum in kidney transplant patients. Culturing of urine samples yielded bluish-green colonies of T. japonicum on Candida chromogenic fungal medium. The isolates were identified as T. japonicum by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI TOF-MS; Autof MS 1000). The identification of T. japonicum was further confirmed by 18S rRNA gene sequencing. In vitro drug susceptibility testing showed that the 2 strains of T. japonicum were resistant to 5-flucytosine, fluconazole, and caspofungin, with dose-dependent sensitivity to itraconazole and voriconazole but sensitivity to amphotericin B. The homology of the 2 T. japonicum strains, as determined by cluster analysis and principal component analysis of MALDI-TOF MS, was ~85%, suggesting a common nosocomial origin. The first 2 case reports of fluconazole-resistant T. japonicum urinary infection in kidney transplant recipients are presented.

Identification of a Novel Linear B Cell Epitope on the Sao Protein of Streptococcus suis Serotype 2.

Surface antigen one (Sao) protein is a bacterial surface protein identified in the important zoonotic pathogen Streptococcus suis serotype 2 (S. suis 2) during an extensive search for functional proteins. The Sao protein is anchored to the bacterial cell wall by the LPVTG motif and is widely distributed in many S. suis serotypes. In this paper, we present the immunodominant epitope peptide of the Sao protein that is recognized by BALB/c antibodies against the Sao protein: 355SEKQMPSVVNENAVTPEKQMTNKENDNIET384 (location Sao355-384). To determine the core epitope recognized by antibodies, we prepared truncation peptide libraries. Analyses of the immunoreactivity of truncation peptides with anti-Sao355-384 serum revealed that the most immunoreactive sequence was 355SEKQMPSVVNENAVTPEK372 (location Sao355-372). Moreover, we observed that this core epitope also showed good specificity based on the ratio of reactivity with serum from S. suis-positive patients compared to serum from S. suis-negative patients. Our results point to the potential of using the Sao355-372 peptide in diagnostic assays to determine S. suis infection in humans.

Identification of a novel alkaline serine protease from gazami crab (Portunus trituberculatus) hepatopancreas and its hydrolysis of myofibrillar protein.

Serine proteases are thought to play a key role in the muscle softening of gazami crab (Portunus trituberculatus) during storage. A serine protease, Pt-sp2, was purified from the hepatopancreas of gazami crab using ammonium sulfate precipitation, anion-exchange and gel filtration chromatography, and was analyzed by mass spectrometry, transcriptome and bioinformatics. It revealed that Pt-sp2 was trypsin-like, with no 100% identical proteins in the NCBI database. The molecular weight of Pt-sp2 was approximately 37.2 kDa. Its optimum pH and temperature were 9.0 and 50 °C, respectively, using t-Butyloxy­carbonyl-Phe-Ser-Arg-4-methyl-coumaryl-7-amide as a substrate. Pt-sp2 was activated in the presence of Ca2+. Both soybean trypsin inhibitor and Nα-Tosyl-l-lysine chloromethyl ketone hydrochloride completely suppressed Pt-sp2 activity, while it was only partially inhibited by phenylmethylsulfonyl fluoride and EDTA. However, PMSF, Pepstatin A and cystatin inhibitor E-64 showed no inhibition on Pt-sp2 protease activity. The Km value of Pt-sp2 was 0.82 µM, and Pt-sp2 effectively hydrolyzed myofibrillar protein at 37 °C.

First report and genetic diversity of porcine bufavirus in China.

BACKGROUND: Bufavirus is a newly discovered zoonotic virus reported in numerous mammals and humans. However, the epidemiological and genetic characteristics of porcine bufaviruses (PBuVs) in China remain unclear. METHODS: To detect PBuVs in China, 384 samples (92 fecal and 292 serum specimens) were collected from 2017 to 2018, covering six provinces in China, and were evaluated by nested PCR. Further, the positive samples from different provinces were selected to obtain the complete genome of Chinese PBuVs. RESULTS: The prevalence rate of PBuV was 16.7% in Chinese domestic pigs in the Guangdong, Guangxi, Fujian, Jiangxi, Anhui, and Henan provinces. Additionally, the positive rate of fecal specimens was higher than that of the serum samples. Next, we sequenced nine near-complete genomes of Chinese field PBuV strains from different provinces. Homology and phylogenetic analyses indicated that Chinese PBuVs have high genetic variation (93.3-99.2%), showed higher nucleotide identity with an Austrian PBuV strain (KU867071.1), and developed into different branches within the same cluster. CONCLUSION: To our knowledge, this is the first report on PBuV in China, expanding the geographic boundaries of PBuV circulation. Our data demonstrate that PBuVs are widely distributed in the six Chinese provinces. Moreover, these Chinese PBuVs exhibit genetic variation and continuous evolution characteristics. Taken together, our findings provide a foundation for future studies on bufaviruses.

Local- and Intermediate-Range Structures on Ordinary and Exotic Phase-Change Materials by Anomalous X-ray Scattering.

Local- and intermediate-range atomic structures were investigated on amorphous phases of an ordinary phase-change material, Ge2Sb2Te5 (GST), and an exotic one, Cu2GeTe3 (CGT), by using anomalous X-ray scattering close to K absorption edges of each element to find a fast amorphous-crystalline phase-change mechanism. The obtained data were analyzed by using reverse Monte Carlo modeling to obtain partial structure factors, partial pair distribution functions, and three-dimensional atomic configurations. Ring statistics were carefully examined to clarify the similarity and difference compared with the corresponding crystal structures, and it was found that amorphous GST has a number of four-membered rings indicating fragments of crystal structure, and amorphous CGT has a remarkable number of three-membered rings showing a collapse of crystal structures composed of purely six-membered rings. A persistent homology analysis was carried out and long-range ring structures of the constituent elements were observed in the amorphous phase, which may originate from fragments of crystal structures with a long-range periodicity.

Identification and genome characterization of a novel feline picornavirus proposed in the Hunnivirus genus.

The genus Hunnivirus, which has been identified in sheep, cattle, and rats, was first proposed in the family Picornaviridae by the International Committee on Taxonomy of Viruses in 2013. In this study, a hunnivirus was detected in fecal samples collected from a diarrheic cat in Southern China in 2017. Genome sequencing and analysis indicated that the novel hunnivirus has the same genome organization as reported for other hunniviruses, 5'UTR-L-P1(VP4-VP2-VP3-VP1)-P2(2A-2B-2C)-P3(3A-3B-3Cpro-3Dpol)-3'UTR, but is genetically divergent. This hunnivirus is proposed as a novel genotype of the species Hunnivirus A and provisionally designated feline hunnivirus. Our study expands the host range of hunnivirus and enriches knowledge on picornaviruses.

Epidemiological features of rabies and molecular evolution characteristics of the rabies virus strains in Xishuangbanna prefecture of Yunnan province, China / 中华实验和临床病毒学杂志

Objective@#To understand the epidemiologic features of the rabies in Xishuang banna prefecture of Yunnan province, China in 2008-2017 and the viral molecular-evolution characteristics.@*Methods@#The data of rabies case questionnaire were collected. The brain tissue samples from mad dogs, suspicious sick dogs and human brain tissue, saliva and cerebrospinal fluid samples from rabies patients were collected in Xishuangbanna. Coding region of nucleoprotein and glycoprotein genes were amplified by RT-PCR and sequenced. Homology and phylogenetic analysis were performed using the relevant bioinformatics software.@*Results@#A total of 62 cases of human rabies were occurred in 28 districts of the 3 counties, Xishuangbanna prefecture in 2008-2017. Of them, 37 cases in Jinghong county, 15 in Menghai county and 10 in Mengla county. In which 48 cases were bitten by domestic dogs (77.42%), 11 cases were bitten by wild dogs (17.74%). Rabies case was occurred every year in the past decade. The seasonal incidence was not obvious. The majority of patients were aged from 30 to 59 years-old, with the youngest 1 year-old and the eldest 91 year-old. The male to female ratio was 1.70∶1, most cases were farmers. The nucleotide sequences of nucleoprotein gene of 9 virus strains (7 from Jinghong, 1 from Menghai and 1 from Mengla) were obtained from the samples of dogs and patients. Homology and phylogenetic analyses indicated that the 5 strains belonged to clade China-Ⅰ, 3 clade China-Ⅱ and 1 clade China-Ⅵ. The nucleotide sequences of glycoprotein gene of 5 virus strains (3 from Jinghong, 1 from Menghai and 1 from Mengla) were obtained from these positive samples, and all were clade China-Ⅰ, it is same with nucleoprotein genes analysis result from these 5 virus strains. These China-Ⅰ and China-Ⅱ strains from Xishuangbanna have a closer genetic relationship with same clade strains isolated from Pu’er and other prefectures of Yunnan province as well as Sichuan, Guizhou and Guangxi. The China-Ⅵ strain from Xishuangbanna share high homology and genetic relationship with China-Ⅵ strains isolated from southwestern Yunnan and neighbouring countries such as Myanmar, Laos and Vietnam in recent years.@*Conclusions@#In Xishuangbanna, rabies mainly occurred in rural area and domestic dog was the main source of transmission. These RABV clades China-Ⅰ, China-Ⅱ and China-Ⅵ were found in this region and the China-Ⅰ was principal clade. The transmission source of China-Ⅰ and China-Ⅱ were from adjacent areas in the province and China-Ⅵ was from Myanmar and Laos.

Semi-analytical solutions for two-dimensional convection-diffusion-reactive equations based on homotopy analysis method.

Convention applied to describe contaminant transport in landfills and groundwater systems is typically characterized by simplified geometries and boundary conditions. As a result, they neglect the more general boundary conditions encountered in the real world, including convection and diffusion of contaminants (e.g., landfill leachate) associated with fluid transportation in the lateral direction. Here, we present semi-analytical solutions that can be used to describe and estimate the contaminants' fate in two-dimensional space. This is achieved by applying the homotopy analysis method (HAM) to create a different order deformation equation series, the sum of which is the solution of the two-dimensional target problem. To ensure the accuracy of the semi-analytical solution, elements of the equation series have been defined and adapted to satisfy the partial differential equation of the discussed problem. Similarly, the convergence of the HAM solution has been achieved by adopting proper convergent control parameters, ensuring the convergence of each element of the deformation equation series. This guarantees that the sum of the equation series is convergent. HAM has been applied to three cases with more general and smooth initial conditions. Good agreement between HAM solutions and numerical solutions from the literature demonstrates the capacity of HAM.

Genomic sequencing and characterization of Theiler's disease-associated virus identified in commercial equine sera in China.

Theiler's disease-associated virus (TDAV) could be the aetiological agent of Theiler's disease. Horses experimentally inoculated with equine plasma containing TDAV develop acute and chronic infections with viraemia. Since its first identification in 2013, TDAV has not been detected in equines in the epidemiological studies that have been conducted. Until now, only one genome sequence of TDAV (HorseA1_serum) had been obtained. In this study, we sequenced the genome of four TDAV strains (A/China, F/China, H/USA and I/USA) in commercial equine sera used for cell culture propagation in China using three rounds of RT-PCR. The PCR primers were designed based on the HorseA1_serum genome sequence. All four TDAV strains had a polyprotein gene that was 9567 nt long, the same nucleotide length as the polyprotein gene of HorseA1_serum. Sequence analysis demonstrated the genetic diversity of TDAV. The nucleotide similarity of the polyprotein genes of the TDAV strains ranged between 90.3 and 93.6 %, with a high amino acid similarity that ranged from 98.2 to 98.8 %. Phylogenetic analysis using the polyprotein gene showed that A/China, F/China, H/USA and I/USA were clustered together with HorseA1_serum in the genus Pegivirus D. This study enriches our knowledge of the genetic diversity of TDAV.

First report and genetic characterization of feline kobuvirus in diarrhoeic cats in China.

Feline kobuvirus (FeKoV) is a newly discovered organism, classified under the species Aichivirus A of the genus Kobuvirus. Since it was first reported in 2013, molecular evidence for FeKoV in the feline population has been restricted to two countries: Korea and Italy. In this study, we collected faecal samples from cats in southern China and detected the FeKoV RNA in these samples. A prevalence rate of 9.9% (8/81) was identified by RT-PCR, and all positive samples were obtained from diarrhoeic animals. In addition, FeKoV was shown positive associated with diarrhoea in cats, with a correlation coefficient of 0.25. Next, we designed three primer pairs with degenerate bases, which targeted the conservative overlapping region of the entire published FeKoV genome, and sequenced the near-complete genome of the first Chinese field FeKoV strain, WHJ-1, using long-fragment PCR. Finally, we analysed WHJ-1's homology and phylogeny using the polyprotein gene. The results indicated that FeKoV has rapidly mutated since it was first discovered. This study will help to better understand FeKoV's epidemiology, evolutionary pattern and genetic diversity.

In silico identification of molecular mimics involved in the pathogenesis of Clostridium botulinum ATCC 3502 strain.

Bacterial pathogens invade and disrupt the host defense system by means of protein sequences structurally similar at global and local level both. The sharing of homologous sequences between the host and the pathogenic bacteria mediates the infection and defines the concept of molecular mimicry. In this study, various computational approaches were employed to elucidate the pathogenicity of Clostridium botulinum ATCC 3502 at genome-wide level. Genome-wide study revealed that the pathogen mimics the host (Homo sapiens) and unraveled the complex pathogenic pathway of causing infection. The comparative 'omics' approaches helped in selective screening of 'molecular mimicry' candidates followed by the qualitative assessment of the virulence potential and functional enrichment. Overall, this study provides a deep insight into the emergence and surveillance of multidrug resistant C. botulinum ATCC 3502 caused infections. This is the very first report identifying C. botulinum ATCC 3502 proteome enriched similarities to the human host proteins and resulted in the identification of 20 potential mimicry candidates, which were further characterized qualitatively by sub-cellular organization prediction and functional annotation. This study will provide a variety of avenues for future studies related to infectious agents, host-pathogen interactions and the evolution of pathogenesis process.