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Goose astrovirus genotype 1 (GAstV-1) has emerged in goose farms in some provinces of China in recent years and is considered to be one of the pathogens of gout in goslings in China. However, few studies have been conducted on the dynamic distribution, tissue tropism, and pathogenesis of GAstV-1 in goslings. In 2022, an epidemiological investigation of goose astrovirus (GAstV) in goslings was conducted in seven provinces of China. During the investigation, a GAstV-1 designated as GAstV-JSXZ was identified in the kidney of an 8-day-old gosling and was successfully isolated from a goose embryo. The full genome sequence of GAstV-JSXZ was determined using the next-generation sequencing technique. The complete genome of GAstV-JSXZ was 7299-nt-long. Interestingly, the phylogenetic analysis revealed that Chinese GAstV-1 has formed two distinct subgroups based on the ORF 2 genomes, designated GAstV-1 1a and GAstV-1 1b. The GAstV-JSXZ shared the highest identity with GAstV-1 1a strain FLX and TZ03 in nucleotides (ORF1a: 98.3–98.4%; ORF1b: 92.3–99.1%; ORF2: 95.8–98.8%) and amino acid sequences (ORF1a: 99.4–99.5%; ORF1b: 98.2–98.8%; ORF2: 97.0–99.4%). To evaluate the pathogenicity of GAstV-1, 1-day-old goslings were inoculated with the virus by oral and subcutaneous injection routes, respectively. The results revealed that the virus causes extensive pathological organ damage, especially in the kidney, liver, and thymus. Virus-specific genomic RNA could be detected in the cloacal swabs and tissues of infected goslings throughout the experiment. The viral copy numbers examined in the kidney and intestine were the highest, followed by the liver and spleen. These results are likely to provide a new understanding of the pathogenicity of GAstV-1 in geese. [ABSTRACT FROM AUTHOR]

Simple Summary: Smoking is the commonest cause of oral cavity squamous cell carcinoma (OC-SCC), but the cause of OC-SCC in nonsmokers is unknown. Our primary goal was to use metagenomic shotgun sequencing (MSS) to directly define the taxonomic composition and functional potential of oral metagenome in nonsmokers with OC-SCC. We found three bacterial phyla, one genus, and one species were enriched in OC-SCC while two phyla, five genera, and 18 species were enriched in controls. Pathways related to metabolism of flavin, biotin, thiamine, heme, sugars, fatty acids, peptidoglycans, and tRNA were more abundant in OC-SCC while those related to metabolism of nucleotides and essential amino acids were more abundant in controls. The MSS method achieved similar results to the commonly used 16S rRNA gene survey in compositional differentiation but differed greatly from prediction using 16S rRNA genes in functional differentiation of microbiomes in OC-SCC and controls. Objectives: Smoking is the commonest cause of oral cavity squamous cell carcinoma (OC-SCC), but the etiology of OC-SCC in nonsmokers is unknown. Our primary goal was to use metagenomic shotgun sequencing (MSS) to define the taxonomic composition and functional potential of oral metagenome in nonsmokers with OC-SCC. Methods: We conducted a case–control study with 42 OC-SCC case and 45 control nonsmokers. MSS was performed on DNA extracted from mouthwash samples. Taxonomic analysis and pathway analysis were done using MetaPhlAn2 and HUMAnN2, respectively. Statistical difference was determined using the Mann–Whitney test controlling false discovery rate. Results: There was no significant difference in age, sex, race, or alcohol consumption between OC-SCC and control patients. There was a significant difference in beta diversity between OC-SCC and controls. At the phylum level, Bacteroidetes and Synergistetes were overly represented in OC-SCC while Actinobacteria and Firmicutes were overly represented in controls. At the genus level, Fusobacterium was overly represented in OC-SCC compared with controls, while Corynebacterium, Streptococcus, Actinomyces, Cryptobacterium, and Selenomonas were overly represented in controls. Bacterial pathway analysis identified overrepresentation in OC-SCC of pathways related to metabolism of flavin, biotin, thiamin, heme, sugars, fatty acids, peptidoglycans, and tRNA and overrepresentation of nucleotides and essential amino acids in controls. Conclusions: The oral microbiome in nonsmoker patients with OC-SCC is significantly different from that of nonsmoker control patients in taxonomic compositions and functional potentials. Our study's MSS findings matched with previous 16S-based methods in taxonomic differentiation but varied greatly in functional differentiation of microbiomes in OC-SCC and controls. [ABSTRACT FROM AUTHOR]

A field isolate (Reo/SDWF /Pheasant/17608/20) of avian orthoreovirus (ARV), isolated from a flock of game-pheasants in Weifang, Shandong Province, was genetically characterized being a field variant or novel strain in our recent research studies in conducting whole genome sequencing by using Next-Generation Sequencing (NGS) technique on Illumina MiSeq platform. Among a total of 870,197 35-151-mer sequencing reads, 297,711 reads (34.21%) were identified as ARV sequences. The de novo assembly of the ARV reads resulted in generation of 10 ARV-related contigs with the average sequencing coverage from 1390× to 1977× according to 10 ARV genome segments. The complete genomes of this pheasant-origin ARV (Reo/SDWF /Pheasant/17608/20) were 23,495 bp in length and consist of 10 dsRNA segments ranged from 1192 bp (S4) to 3958 bp (L1) encoding 12 viral proteins. Sequence comparison between the SDWF17608 and classic ARV reference strains revealed that 58.1–100% nucleotide (nt) identities and 51.4–100% amino acid (aa) identities were in genome segment coding genes. The 10 RNA segments had conversed termini at 5' (5'-GCUUUU) and 3' (UCAUC-3') side, which were identical to the most published ARV strains. Phylogenetic analysis revealed that this pheasant ARV field variant was closely related with chicken ARV strains in 7 genome segment genes, but it possessed significant sequence divergence in M1, M3 and S2 segments. These findings suggested that this pheasant-origin field variant was a divergent ARV strain and was likely originated from reassortments between different chicken ARV strains. [ABSTRACT FROM AUTHOR]

Background: Tuberculosis (TB) seriously threatens individual and public health. Recently, TB outbreaks in schools have been reported more frequently in China and have attracted widespread attention. We reported three TB outbreaks in high schools in Hunan Province, China. Methods: When a tuberculosis patient was reported in a school, we carried out field epidemiological investigations, including tuberculin skin testing (TST), chest X-ray (CXR) and laboratory test for all close contacts, and whole-genome sequencing (WGS) analyses to understand the transmission patterns, the causes and the risk factors for the outbreaks, thereby providing a foundation for the control of TB epidemics in schools. Results: A total of 49 students with TB patients were identified in the three schools where TB outbreaks occurred, including nine patients in School A, 14 patients in School B, and 26 patients in School C. In Schools A, B and C, the putative attack rates in the classes of the index case were 13.8% (8/58), 7.6% (5/66), and 40.4% (21/52), while the putative attack rates of expanding screening in the school were 0.3% (1/361), 0.2% (9/3955), and 0.2% (5/2080), respectively. Thirteen patients had patient delay, with a median delay interval of 69 days (IQR 30.5– 113 days). Twelve patients had a healthcare diagnostic delay with a median delay interval of 32 days (IQR 24– 82 days). Phylogenetic analysis of culture-positive patients revealed that most of them shared a small genetic distance (≤ 12 SNPs), with three separate genetic clusters (including one MDR-TB genomic cluster), indicating the recent transmission of Mycobacterium tuberculosis strains. Conclusion: This combination of field investigation and WGS analysis revealed the transmission of three TB outbreaks in schools. Reinforced implementation is needed to improve timely case finding and reduce diagnosis delay in routine TB control in the school population. [ABSTRACT FROM AUTHOR]

Since the first report from Cherry Valley ducks on a commercial duck farm in China (2008), duck astrovirus type 1(DAstV‐1)‐associated duck viral hepatitis (DVH) have been detected in several commercial duck flocks. A highly acute disease characterized by hepatitis broke out in ducklings in Shandong Province in March 2021, all diseased ducks have been immunized against the duck viral hepatitis vaccine. One DAstV‐1 strain, designated as DAstV‐SDWF, was isolated from a diseased duckling. Here, the isolation, cultivation, and characterization of DAstV‐1 isolate are described. The isolated astrovirus grew well in the LMH cell line. To determine the entire genomic of the DAstV‐SDWF isolate, the next‐generation sequencing (NGS) technique was conducted on the Illumina HiSeq platform. Complete genome sequence analysis revealed that DAstV‐SDWF isolate was 91.6%–98.6% homology with others DAstV‐1 deposited in Genbank. Similar clinical symptoms were successfully reproduced by an experimental infection study using the DAstV‐SDWF isolate. DAstV‐SDWF is the first DAstV‐1 strain whose experimental infection study has been conducted in China. Results of the above data demonstrated that DAstV‐1 could be one of the causative agents of the DVH occurring in China. The present works are likely to provide new insights into the pathogenicity and evolution of DAstV‐1 in ducks. [ABSTRACT FROM AUTHOR]

Since 2013, there have been an increasing number of cases of arthritis in broilers caused by avian orthoreovirus (ARV) in China, and the virus remains highly virulent in chicks with high‐level maternal antibodies. However, little information is available about the complete gene analysis and pathogenicity of the epidemic ARVs. In the study, the ARV strain (V‐ARV‐SD26) was isolated from broilers associated with arthritis in Shandong Province. To genetically characterize the ARV strain, the whole‐genome sequencing was conducted by next‐generation sequencing (NGS) technique. Sequence analysis demonstrated that V‐ARV‐SD26 might have acquired its current genomic composition through several homologous and, in case of the λC, μA and σB, divergent reassortment events. To further investigate the pathogenicity of the strain, 160 one‐day‐old Ross broilers with maternal antibodies were equally divided into four groups (foodpad‐, eye mucosa‐ and intramuscular‐inoculated groups and the negative control group), three experimental groups were inoculated separately with the low‐dose virus fluid, and the negative control was equally inoculated with sterile PBS. The results showed that the symptoms of broilers in foodpad inoculation group were the most serious, while that of the eye mucosa infection group were the mildest. Meanwhile, the cloacal cotton swabs and organs were collected for qRT‐PCR detection to evaluate the infection status. In conclusion, these findings indicate that V‐ARV‐SD26 is a divergent ARV strain, which provide experimental data for the prevention and control of newly emerged reovirus, and have a certain reference value for the preparation and evaluation of new vaccines. [ABSTRACT FROM AUTHOR]

Clostridioides difficile sequence type 2 (ST2) has been increasingly recognized as one of the major genotypes in China, while the genomic characteristics and biological phenotypes of Chinese ST2 strains remain to be determined. We used whole-genome sequencing and phylogenetic analysis to investigate the genomic features of 182 ST2 strains, isolated between 2011 and 2017. PCR ribotyping (RT) was performed, and antibiotic resistance, toxin concentration, and sporulation capacity were measured. The core genome Maximum-likelihood phylogenetic analysis showed that ST2 strains were distinctly segregated into two genetically diverse lineages [L1 (67.0% from Northern America) and L2], while L2 further divided into two sub-lineages, SL2a and SL2b (73.5% from China). The 36 virulence-related genes were widely distributed in ST2 genomes, but in which only 11 antibiotic resistance-associated genes were dispersedly found. Among the 25 SL2b sequenced isolates, RT014 (40.0%, n = 10) and RT020 (28.0%, n = 7) were two main genotypes with no significant difference on antibiotic resistance (χ2 = 0.024–2.667, P > 0.05). A non-synonymous amino acid substitution was found in tcdB (Y1975D) which was specific to SL2b. Although there was no significant difference in sporulation capacity between the two lineages, the average toxin B concentration (5.11 ± 3.20 ng/μL) in SL2b was significantly lower in comparison to those in L1 (10.49 ± 15.82 ng/μL) and SL2a (13.92 ± 2.39 ng/μL) (χ2 = 12.30, P < 0.05). This study described the genomic characteristics of C. difficile ST2, with many virulence loci and few antibiotic resistance elements. The Chinese ST2 strains with the mutation in codon 1975 of the tcdB gene clustering in SL2b circulating in China express low toxin B, which may be associated with mild or moderate C. difficile infection. [ABSTRACT FROM AUTHOR]

This study was the first report complete chloroplast genome of Nouelia insignis (Asteraceae, Hyalideae), the large shrubs to small trees endemic to China. The circular whole cp genome of N. insignis was 151,524 bp in length, containing a large single-copy (LSC) region of 83,145 bp and a small single-copy (SSC) region of 18,261 bp. These two regions were separated by a pair of inverted repeat regions (IRa and IRb), each of them 25,060 bp in length. A total of 135 functional genes were encoded, consisting of 89 protein-coding genes, 38 tRNA genes, and eight rRNA genes. The overall GC content of the chloroplast genome sequence was 37.8%, and the GC contents of the LSC, SSC, and IR regions were 35.9, 31.5, and 43.2%, respectively. The phylogenetic analysis by the Bayesian analysis showed that the species of N.insignis was sister group with Gerbera jamesonii by strong support values, and thus was closely related to members of subfamilies of Cichorioideae and Pertyoideae. These results will be useful for the future studies of Asteraceae in the worldwide. [ABSTRACT FROM AUTHOR]

Since December 2017, an infectious disease has caused economic hardship for duck farms and breeding ducks in many regions of China. This disease characterized by spleen necrosis and swelling, is due to a variant strain of duck orthoreovirus (DRV) (Duck/N‐DRV‐XT18/China/2018), which we isolated from the spleen of diseased ducks. After isolating the virus, we used next‐generation sequencing technology to determine the entire genomic of the virus. Our phylogenetic analysis of 10 genomic segments showed that the N‐DRV‐XT18 strain is closely related to orthoreovirus isolates derived from ducks and geese, with nucleotide sequence identities for 10 genomic fragments ranging between 49.8% and 99.3%. In contract, the nucleotide sequence of N‐DRV‐XT18 genomic fragments are only 38.6% to 78.8% similar to the chicken orthoreovirus isolate. Therefore, we determined that this pathogen, causing duck spleen necrosis, is a new variant of a duck orthoreovirus that is significantly different from any previously reported waterfowl‐derived othoreovirus. [ABSTRACT FROM AUTHOR]

Ducks infected with duck circovirus (DuCV) show symptoms such as feather loss, growth retardation and low body weight in the flock. The virus induces immunosuppression and increases the prevalence of infection with other pathogens. However, most studies on duck circovirus were focused on coinfection, and fewer studies had been conducted on the pathogenicity of duck circovirus alone. The aim of the present study was to investigate the pathogenesis of DuCV-1 in experimentally infected specific pathogen-free ducks. In this study, we sequenced the whole genome of a strain of duck circovirus and identified the virus genotype as DuCV-1b. This strain of duck circovirus was named SDLH(OR567883). Animal pathogenicity experiments were then conducted, wherein specific pathogen-free ducks were infected by mucosal injection and abdominal injection. Infected ducks were sampled for 4 consecutive weeks after infection and showed symptoms of dwarfism. We further examined the replication of DuCV-1 in the ducks. The highest virus titers in the 2 infection groups were found in the liver and spleen, with different results for the different routes of infection. Pathological sections of duck organs were made and it was found that organs such as the liver and spleen were damaged by DuCV-1. In conclusion, our experimental results indicate that DuCV-1 can infect ducks individually and cause widespread organ damage in infected ducks. [ABSTRACT FROM AUTHOR]

A fowl adenovirus variant designated as DAdV-JSXZ strain was isolated from the tissue specimen of fallopian tubes of a duck case, which was submitted from a 276-day-old Cherry valley breeding duck flock experienced egg-dropping syndromes in March 2022. Full-genome sequence of the DAdV-I JSXZ strain by next-generation sequencing revealed that the complete genome length of DAdV-JSXZ strain was 33,213 nucleotides and shared a high degree of nucleotide identity (97.0–99.4 %) with other DAdV-I reference strains. In pathogenicity studies, this isolated duck JSXZ strain reproduced similar egg-dropping symptoms in healthy breeding ducks, pathologic lesions of follicular hemorrhage, and the laid eggs in low fertilization and hatchability rates. Our research findings demonstrated that DAdV-I JSXZ strain was one of the causative agents of duck egg dropping syndrome in egg-laying ducks and could cause acute respiratory symptoms in ducklings. • This is the first time to report the complete viral genome of DAdV-1 in breeding ducks. • This study provides scientific evidence that DAdV-1 could be one of the pathogens of duck egg drop. • This study provides evidence of possible vertical transmission of DAdV-1. [ABSTRACT FROM AUTHOR]

• A total of 147 S. Enteritidis were isolated from a poultry breeding enterprise. • All S. Enteritidis belonged to ST11, and further divided into 4 rSTs and 64 cgSTs. • SNPs typing suggested the presence of vertical transmission of S. Enteritidis. • Heterogeneous antimicrobial resistance was present in clonally related strains. • One bla CTX-M-14 -carrying plasmid and two virulence-resistance plasmids were characterized. Human salmonellosis caused by the consumption of eggs and chicken meat contaminated with Salmonella Enteritidis has become a continuing public health concern worldwide. In this study we adopted whole genome sequencing (WGS) to determine the genetic relationship and antimicrobial resistance of S. enterica strains isolated from a poultry breeding enterprise that consists of one breeding chicken farm, one egg hatchery and one commercial chicken farm. A total of 148 S. enterica including 147 S. Enteritidis strains were isolated from 2100 fecal swab samples, with 16 (5.3 %, 16/300) from breeding chicken farm, 38 (4.2 %, 38/900) from egg hatchery and 94 (10.4 %, 94/900) from commercial chicken farm. WGS revealed that all 147 S. Enteritidis strains belonged to ST11, and further divided into 4 different ribosomal STs and 64 core genome STs. Single nucleotide polymorphism typing suggested the presence of the vertical transmission of S. Enteritidis from breeding chicken to commercial chicken. Three different antimicrobial-resistant plasmids including one bla CTX-M-14 -carrying plasmid and two virulence-resistance plasmids were characterized, resulting in the heterogeneous antimicrobial resistance of clonally related S. Enteritidis strains. Routine surveillance in breeding chicken farms is conducive to the control of S. Enteritidis from farm to fork. [ABSTRACT FROM AUTHOR]