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Astroviruses are an emerging hazard in hatchery management, and they are responsible for great economic losses in poultry farms worldwide. Chicken astrovirus (CAstV) and avian nephritis virus (ANV) are involved in decreased hatching rate, mainly due to embryo deaths, and pale and runted chicks too weak to hatch. Hatchery diseases occur through direct egg infection or vertical transmission. However, the specific role of astroviruses in these diseases is still largely unknown. To elucidate this a high-quality Piedmont hatchery (Italy) was selected based on farm clinical signs and the astrovirus serological positivity in the breeder farm. The viral dynamics were followed from the hatchery to the farm. The presence of both CAstV and ANV was monitored by evaluating the viral load in environmental and bird samples. Interestingly, both viruses were detected in chick yolk sacs at 18 days of incubation, and in the gut contents of 1-day chicks, whereas in the hatchery environmental samples, the eggs' external surfaces and the embryos' intestinal contents were negative. The CAstV copy number was higher in bird tissues from the hatchery, while a greater ANV load was detected in bird tissues from the farm. Moreover, our data suggest vertical transmission of both viruses. Frequent co-infection was also observed in samples from both the hatchery and the farm. Lastly, the first Italian full-length genome and molecular characterization of ANV was obtained. RESEARCH HIGHLIGHTS: Identified the role of the hatchery in astrovirus transmission. Sequenced the avian nephritis virus complete genome. Investigated tissue distribution of astrovirus from egg to chicks. Demonstrated co-infection of ANV/CAstV. [ABSTRACT FROM AUTHOR]

Background: The garden dormouse (Eliomys quercinus) has experienced a significant population decline across Europe in recent decades. While habitat loss and climate change are often cited as primary factors, pathogen exposure, either to novel or to previously known, may play a role in such a decline. This study aimed to investigate the presence of polyomaviruses in garden dormice, given that these viruses are highly prevalent and can cause disease, particularly in immunocompromised individuals. Methods: The carcasses of garden dormice (n = 89) were collected throughout Germany. Kidney samples were tested for the presence of polyomavirus DNA using nested degenerate and specific diagnostic PCRs. Seroprevalence was assessed from chest cavity fluid samples through an enzyme-linked immunosorbent assay using polyomavirus VP1 virus-like particles produced in yeast. Results: A new polyomavirus, related to chimpanzee (Pan troglodytes) polyomaviruses 4 and 5 and human Merkel cell polyomavirus, was identified in the garden dormouse. Two 5,380 bp-length complete viral genomes were sequenced from dormice kidney samples (sequences PQ246041 and PQ246042). Genes encoding the putative structural proteins VP1, VP2, and VP3, as well as the Large, Middle, and small T antigens, containing conserved functional domains were identified. Polyomavirus DNA was detected in 2 of 74 dormice (2.7%, 95% confidence interval: 0-6.4%) through PCR, while 12 of 69 animals (17.4%, 95% confidence interval: 8.4-26.3%) tested positive for polyomavirus-specific antibodies. Conclusions: In conclusion, here we describe a novel polyomavirus in the garden dormouse with molecular and serological detection. Pairwise sequence comparison and phylogenetic analysis suggest that this novel virus may represent a novel species within the genus Alphapolyomavirus. Future work should examine if this virus is garden dormouse-specific and whether it is associated with disease in dormice. [ABSTRACT FROM AUTHOR]

Rotaviruses are well‐recognized pathogens responsible for diarrhea in humans and various animal species, with Rotavirus A the most often detected and most thoroughly described. Rotaviral disease is an important concern in pathology of pigeons as well, as pigeon rotavirus A was proven to play a major role in young pigeon disease (YPD). However, rotaviruses of other groups have been so far understudied in birds. This paper describes the first finding of Rotavirus G in domestic pigeon in Europe, as well as the recovery of three complete genomes of pigeon rotavirus A with Oxford Nanopore Sequencing. Quantification of pigeon rotavirus A genetic material with droplet digital polymerase chain reaction (PCR) in pigeons suffering from diarrhea and in asymptomatic pigeons was also performed in the frameworks of this study and resulted in determination of statistically highly significant differences between the groups in both detection rate and shedding of the virus. Phylogenetic analysis revealed the close relationship of acquired strains with those originating from pigeons from Europe, North America, Asia, and Australia, indicating a broad geographical spread of pigeon rotaviruses. Results of our research shed more light on occurrence and diversity of Rotavirus species in pigeons. [ABSTRACT FROM AUTHOR]

Background: Rotaviruses are the major etiological agents of gastroenteritis and diarrheal outbreaks in plenty of mammalian species. The genus Rotavirus is highly diverse and currently comprises nine genetically distinct species, and four of them (A, B, C, and H) are common for humans and pigs. There is a strong necessity to comprehend phylogenetic relationships among rotaviruses from different host species to assess interspecies transmission, specifically between humans and livestock. To reveal the genetic origin of rotaviruses from Russian pig farms, nanopore-based metagenomic sequencing was performed on the PCR-positive specimens. Methods: Samples were selected among the cases submitted to routine diagnostic or monitoring studies to the Laboratory of Biochemistry and Molecular Biology of "Federal Scientific Center VIEV" (Moscow, Russia). The selected positive samples were genotyped using nanopore sequencing method. Results: Five porcine RVA isolates were completely sequenced, and genotype analysis revealed various porcine G/P genogroups: G2, G3, G4, G5, G11 and P[6], P[7], P[13], P[23], P[27] with a typical backbone constellation I5-R1-C1-M1-A8-N1-T1/7-E1-H1. The RVB isolate was detected in combination with RVA in a rectal swab from a diseased pig in Krasnoyarsk Krai. It was characterized by the following genogroups: G15-P[X]-I11-R4-C4-M4-A8-N10-T4-E4-H7. The first complete porcine RVC genome from Russia was obtained with genomic constellation G6-P[5]-I14-R1-C1-M1-A7-N9-T6-E1-H1, and the phylogenetic analysis revealed putative novel genotype group for the VP6 gene-I14. Additionally, the first porcine kobuvirus isolate from Russia was phylogenetically characterized. Conclusions: The applied nanopore sequencing method successfully genotyped the RV isolates and additionally revealed co-circulated species. The study demonstrates high genetic variability of Russian RVA isolates in VP4/VP7 genes and phylogenetically describes local RVB and RVC. Complete characterization of genomic segments is a crucial methodology in tracing the rotavirus's evolution and evaluating interspecies transmissions. [ABSTRACT FROM AUTHOR]

The pathogenic watermelon mosaic virus (WMV) causes severe crop losses and threatens cucurbits production worldwide. This study presents the first sequenced genome of WMV from Iraq and the second from Iran. The near-complete genome sequences of Iranian (In) and Iraqi (Iq) isolates were, respectively, 9999 and 10,041 nucleotides in length, excluding the 3′-terminal poly(A) tail. WMV-Iq and WMV-In shared nucleotide identities of 92.61–96.32% with those of 30 worldwide representative WMV isolates in GenBank and identities of 93.04–93.90% with a previously reported Iranian isolate IR02-54. Phylogenetic analysis based on the genomic sequences clustered all 32 isolates into seven groups (G1–G7), in which WMV-Iq and WMV-In clustered alongside 12 isolates from Asia, Europe, and America in group G1. Phylogenetic analysis of nucleotide sequences of the coat protein gene of WMV isolates resolved into five phylogroups (I–V) and both isolates, WMV-Iq and WMV-In, clustered together with isolates in group I and contain the N-terminal KEA motif, which is the characteristic of group I isolates. Our results show that WMV-Iq and WMV-In are recombinant isolates with multiple recombination events detected across the genome. Symptom intensity of WMV isolates in experimental hosts was variable and depended upon the host species, virulence of isolates, and environmental conditions. [ABSTRACT FROM AUTHOR]

A rhizosphere strain, Achromobacter insolitus LCu2, was isolated from alfalfa (Medicago sativa L.) roots. It was able to degrade of 50% glyphosate as the sole phosphorus source, and was found resistant to 10 mM copper (II) chloride, and 5 mM glyphosate–copper complexes. Inoculation of alfalfa seedlings and potato microplants with strain LCu2 promoted plant growth by 30–50%. In inoculated plants, the toxicity of the glyphosate–copper complexes to alfalfa seedlings was decreased, as compared with the noninoculated controls. The genome of A. insolitus LCu2 consisted of one circular chromosome (6,428,890 bp) and encoded 5843 protein genes and 76 RNA genes. Polyphasic taxonomic analysis showed that A. insolitus LCu2 was closely related to A. insolitus DSM23807T on the basis of the average nucleotide identity of the genomes of 22 type strains and the multilocus sequence analysis. Genome analysis revealed genes putatively responsible for (1) plant growth promotion (osmolyte, siderophore, and 1-aminocyclopropane-1-carboxylate deaminase biosynthesis and auxin metabolism); (2) degradation of organophosphonates (glyphosate oxidoreductase and multiple phn clusters responsible for the transport, regulation and C–P lyase cleavage of phosphonates); and (3) tolerance to copper and other heavy metals, effected by the CopAB–CueO system, responsible for the oxidation of copper (I) in the periplasm, and by the efflux Cus system. The putative catabolic pathways involved in the breakdown of phosphonates are predicted. A. insolitus LCu2 is promising in the production of crops and the remediation of soils contaminated with organophosphonates and heavy metals. [ABSTRACT FROM AUTHOR]

Objectives: The pathogen of Pantoea stewartii (Ps) is the causal agent of bacterial disease in corn and various graminaceous plants. Ps has two subspecies, Pantoea stewartii subsp. stewartia (Pss) and Pantoea stewartii subsp. indologenes (Psi). This study presents two complete genomes of Ps strains including ATCC 8199 isolated from maize and PSCN1 causing bacterial wilt in sugarcane. The two bacterial genomes information will be helpful for taxonomy analysis in this genus Pantoea at whole-genome levels and accurately discriminated the two subspecies of Pss and Psi. Data description: The reference strain ATCC 8199 isolated from maize was purchased from Beijing Biobw Biotechnology Co., Ltd. (China) and the strain of PSCN1 was isolated from sugarcane cultivar YZ08-1095 in Zhanjiang, Guangdong province of China. Two complete genomes were sequenced using Illumina Hiseq (second-generation) and Oxford Nanopore (third-generation) platforms. The genome of the strain ATCC 8199 comprised of 4.78 Mb with an average GC content of 54.03%, along with five plasmids, encoding a total of 4,846 gene with an average gene length of 827 bp. The genome of PSCN1 comprised of 5.03 Mb with an average GC content of 53.78%, along with two plasmids, encoding a total of 4,725 gene with an average gene length of 913 bp. The bacterial pan-genome analysis highlighted the strain ATCC 8199 was clustered into a subgroup with a Pss strain CCUG 26,359 from USA, while the strain PSCN1 was clustered into another subgroup with a Ps strain NRRLB-133 from USA. These findings will serve as a useful resource for further analyses of the evolution of Ps strains and corresponding disease epidemiology worldwide. [ABSTRACT FROM AUTHOR]

The Back-breasted Thrush (Turdus dissimilis Blyth 1847), a medium-sized Turdus bird in the Turdidae family, is widely distributed in montane areas from northeastern India and Myanmar to southern China. The mitochondrial DNA of T. dissimilis is packaged in a compact 16,761-basepair (bp) circular molecule with A + T content of 52.50%. It contains 37 typical mitochondrial genes, including 13 protein-coding genes, 2 rRNAs and 22 tRNAs, and 1 noncoding region. We reconstructed a phylogenetic tree based on the mitogenome sequences of 10 Turdidae species and one outgroup. Phylogenetic analysis indicated that T. dissimilis is a sister taxon to T. unicolor. The new mitogenome data would provide useful information for application in conservation. [ABSTRACT FROM AUTHOR]

Abstract Background The garden dormouse (Eliomys quercinus) has experienced a significant population decline across Europe in recent decades. While habitat loss and climate change are often cited as primary factors, pathogen exposure, either to novel or to previously known, may play a role in such a decline. This study aimed to investigate the presence of polyomaviruses in garden dormice, given that these viruses are highly prevalent and can cause disease, particularly in immunocompromised individuals. Methods The carcasses of garden dormice (n = 89) were collected throughout Germany. Kidney samples were tested for the presence of polyomavirus DNA using nested degenerate and specific diagnostic PCRs. Seroprevalence was assessed from chest cavity fluid samples through an enzyme-linked immunosorbent assay using polyomavirus VP1 virus-like particles produced in yeast. Results A new polyomavirus, related to chimpanzee (Pan troglodytes) polyomaviruses 4 and 5 and human Merkel cell polyomavirus, was identified in the garden dormouse. Two 5,380 bp-length complete viral genomes were sequenced from dormice kidney samples (sequences PQ246041 and PQ246042). Genes encoding the putative structural proteins VP1, VP2, and VP3, as well as the Large, Middle, and small T antigens, containing conserved functional domains were identified. Polyomavirus DNA was detected in 2 of 74 dormice (2.7%, 95% confidence interval: 0-6.4%) through PCR, while 12 of 69 animals (17.4%, 95% confidence interval: 8.4-26.3%) tested positive for polyomavirus-specific antibodies. Conclusions In conclusion, here we describe a novel polyomavirus in the garden dormouse with molecular and serological detection. Pairwise sequence comparison and phylogenetic analysis suggest that this novel virus may represent a novel species within the genus Alphapolyomavirus. Future work should examine if this virus is garden dormouse-specific and whether it is associated with disease in dormice.

Abstract Background Rotaviruses are the major etiological agents of gastroenteritis and diarrheal outbreaks in plenty of mammalian species. The genus Rotavirus is highly diverse and currently comprises nine genetically distinct species, and four of them (A, B, C, and H) are common for humans and pigs. There is a strong necessity to comprehend phylogenetic relationships among rotaviruses from different host species to assess interspecies transmission, specifically between humans and livestock. To reveal the genetic origin of rotaviruses from Russian pig farms, nanopore-based metagenomic sequencing was performed on the PCR-positive specimens. Methods Samples were selected among the cases submitted to routine diagnostic or monitoring studies to the Laboratory of Biochemistry and Molecular Biology of “Federal Scientific Center VIEV” (Moscow, Russia). The selected positive samples were genotyped using nanopore sequencing method. Results Five porcine RVA isolates were completely sequenced, and genotype analysis revealed various porcine G/P genogroups: G2, G3, G4, G5, G11 and P[6], P[7], P[13], P[23], P[27] with a typical backbone constellation I5-R1-C1-M1-A8-N1-T1/7-E1-H1. The RVB isolate was detected in combination with RVA in a rectal swab from a diseased pig in Krasnoyarsk Krai. It was characterized by the following genogroups: G15-P[X]-I11-R4-C4-M4-A8-N10-T4-E4-H7. The first complete porcine RVC genome from Russia was obtained with genomic constellation G6-P[5]-I14-R1-C1-M1-A7-N9-T6-E1-H1, and the phylogenetic analysis revealed putative novel genotype group for the VP6 gene-I14. Additionally, the first porcine kobuvirus isolate from Russia was phylogenetically characterized. Conclusions The applied nanopore sequencing method successfully genotyped the RV isolates and additionally revealed co-circulated species. The study demonstrates high genetic variability of Russian RVA isolates in VP4/VP7 genes and phylogenetically describes local RVB and RVC. Complete characterization of genomic segments is a crucial methodology in tracing the rotavirus's evolution and evaluating interspecies transmissions.

Abstract Objectives The pathogen of Pantoea stewartii (Ps) is the causal agent of bacterial disease in corn and various graminaceous plants. Ps has two subspecies, Pantoea stewartii subsp. stewartia (Pss) and Pantoea stewartii subsp. indologenes (Psi). This study presents two complete genomes of Ps strains including ATCC 8199 isolated from maize and PSCN1 causing bacterial wilt in sugarcane. The two bacterial genomes information will be helpful for taxonomy analysis in this genus Pantoea at whole-genome levels and accurately discriminated the two subspecies of Pss and Psi. Data description The reference strain ATCC 8199 isolated from maize was purchased from Beijing Biobw Biotechnology Co., Ltd. (China) and the strain of PSCN1 was isolated from sugarcane cultivar YZ08-1095 in Zhanjiang, Guangdong province of China. Two complete genomes were sequenced using Illumina Hiseq (second-generation) and Oxford Nanopore (third-generation) platforms. The genome of the strain ATCC 8199 comprised of 4.78 Mb with an average GC content of 54.03%, along with five plasmids, encoding a total of 4,846 gene with an average gene length of 827 bp. The genome of PSCN1 comprised of 5.03 Mb with an average GC content of 53.78%, along with two plasmids, encoding a total of 4,725 gene with an average gene length of 913 bp. The bacterial pan-genome analysis highlighted the strain ATCC 8199 was clustered into a subgroup with a Pss strain CCUG 26,359 from USA, while the strain PSCN1 was clustered into another subgroup with a Ps strain NRRLB-133 from USA. These findings will serve as a useful resource for further analyses of the evolution of Ps strains and corresponding disease epidemiology worldwide.

The Back-breasted Thrush (Turdus dissimilis Blyth 1847), a medium-sized Turdus bird in the Turdidae family, is widely distributed in montane areas from northeastern India and Myanmar to southern China. The mitochondrial DNA of T. dissimilis is packaged in a compact 16,761-basepair (bp) circular molecule with A + T content of 52.50%. It contains 37 typical mitochondrial genes, including 13 protein-coding genes, 2 rRNAs and 22 tRNAs, and 1 noncoding region. We reconstructed a phylogenetic tree based on the mitogenome sequences of 10 Turdidae species and one outgroup. Phylogenetic analysis indicated that T. dissimilis is a sister taxon to T. unicolor. The new mitogenome data would provide useful information for application in conservation.

Treponema pedis, a fastidious anaerobic spirochete, is one of the main pathogens involved in the development and progression of bovine digital dermatitis (BDD), a lameness-causing hoof infection in cattle. Here, the complete genome sequencing of T. pedis GNW45 isolated from a dairy cow infected with BDD, was presented. Libraries for long and short reads were sequenced using PacBioRSII and Illimuna HiSeqXTen platforms, respectively. De-novo assembly was done using the long reads, producing a circular contig, by which the short reads were aligned to generate a more accurate genome sequence. The genome has a total size of 3,077,465 base pairs, with 36.84% guanine-cytosine content. A total of 2,749 protein-coding sequences, seven ribosomal RNA’s, and 45 transfer RNA’s were annotated. Functional analysis revealed genes associated with pathogenicity and survivability in the complex pathobiome of BDD. This study provided novel insights into the survival and pathogenic mechanisms of T. pedis GNW45.

Objectives: The objective of this study was to investigate the resistance mechanisms of a multidrug-resistant Salmonella Kentucky ST198 FJ-2064 isolated from a patient in China. Methods: The antimicrobial susceptibility of FJ-2064 was determined by the standard disc dilution and broth microdilution methods. The complete genome of FJ-2064 was sequenced using PacBio and Illumina MiSeq platforms. Polymerase chain reaction (PCR) and S1-PFGE were utilized to confirm the mutation sites and the genomic plasmids, respectively. Results: Isolate FJ-2064 belongs to sequence type ST198 and harboured no visible large plasmids, but was concurrent resistant to 22 detected antimicrobial agents including cefotaxime, ciprofloxacin, and azithromycin. The complete genome sequence identified 20 acquired antibiotic resistance genes (ARGs) and five chromosomal mutations in the gyrA and parC genes of the quinolone resistance determining regions (QRDRs) in FJ-2064. In addition, PCR sequencing confirmed that most of the ARGs were clustered on one multidrug-resistant region and a variant of SGI1-K. In particular, the bla-TEM-1 and bla-CTX−M-55, qnrS1, mph(A) genes, which confer resistance to cephalosporins, quinolones, and macrolides respectively, were all located on the multidrug-resistant region. Conclusions: We have demonstrated one multidrug-resistant region and a variant of SGI1-K in a Salmonella Kentucky ST198 that is co-resistant to cefotaxime, ciprofloxacin, and azithromycin.

Bacillus subtilis, a gram-positive bacterium commonly found in soil, is an excellent organism for plant biocontrol. B. subtilis KRS015, an endophyte isolated from the seed of Gossypium hirsutum ‘Zhongzhimian No. 2’, has been proven based on its antagonistic activity against diverse fungal pathogens of plants, including Verticillium dahliae, a fungal pathogen of Verticillium wilt in various plants such as cotton. Here, we report the complete genome sequence of B. subtilis KRS015. The size of the KRS015 genome is 4,331,506 bp. Fifteen gene clusters for antibiotics and secondary metabolites and 412 genes involved in plant–bacterial interactions were identified in this genome. These findings qualified the potential of B. subtilis KRS015 as a biocontrol agent and may help reveal the molecular basis of its antagonistic mechanisms. [Figure: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Cercospora beticola, causal agent of Cercospora leaf spot of sugarbeet (Beta vulgaris subsp. vulgaris), is the most economically damaging foliar pathogen inhibiting sugarbeet production. Despite the global importance of C. beticola, significant knowledge gaps exist pertaining to pathogen virulence and adaptation to management practices that include the use of resistant host varieties and the application of different fungicide chemistries. The lack of a high-quality reference genome for C. beticola has hindered research progression addressing these critical knowledge gaps. Here, we report the complete and gapless genome assembly and annotation of the reference C. beticola isolate Cb09-40. We identified misassembled regions in the previous reference genome and annotated previously missing protein-coding genes. This work establishes a high-quality reference genome that can serve as the foundation for future comparative genomic, population genetic, and evolutionary studies into the virulence and evolution of this important plant pathogen. [Figure: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Bagaza virus (BAGV) is a flavivirus that affects avian species. In Europe, it was detected for the first time in Spain in 2010, exhibiting high genetic relatedness to Israel turkey meningoencephalomyelitis virus (ITMV) isolates from Israel. After a period of epidemiological silence, BAGV re-emerged, causing important outbreaks in 2019 and 2021. This study aims to characterize the newly detected strains and to elucidate if these recent outbreaks were caused by single or different virus introductions into the country. Hence, Spanish BAGV isolates from 2019 (n = 3) and 2021 (n = 1) outbreaks, obtained from red-legged partridges in Cádiz, were sequenced and further characterized. The phylogenetic analyses showed that they belong to two different genotypes: BAGV-Genotypes 1 and 2. Isolates from 2019 belong to BAGV-Genotype 1, closely related to isolates from Senegal, where BAGV has been circulating for decades. In turn, the 2021 isolates belong to BAGV-Genotype 2, closely related to those detected in Spain in 2010. Additionally, the comparison of the viral polyproteins of several BAGV isolates from both genotypes supports and confirms the phylogenetic findings. To conclude, BAGV has been introduced into Spain on at least three independent occasions, with alternating genetic clades, thus confirming that BAGV is able to sporadically reach Southern Europe. [ABSTRACT FROM AUTHOR]

Avian encephalomyelitis (AE) is an important infectious poultry disease worldwide that is caused by avian encephalomyelitis virus (AEV). The causative virus can be transmitted both horizontally and vertically. In the present study, an AEV suspected outbreak with typical neurological signs occurred in broilers. Histopathological examination, RT-PCR assay and full genome sequencing were applied to confirm the presence of AEV. Phylogenetic analysis of the full genome sequence showed that the detected AEV strain at 7055 nucleotide length is classified in cluster I and is closely related to vaccinal USA and China originated isolates. Although, the outbreaks of AE in progeny of vaccinated breeders have been reported previously, the source of infection was unknown. Based on the results obtained in this study, the outbreaks are vaccine-originated. This study provides the first whole genome analysis of AEV from Iran and reveals that the AEV possesses a hepatitis C virus-like internal ribosome entry site. [ABSTRACT FROM AUTHOR]

Porcine circoviruses (PCVs) are widely distributed in swine herds. PCV2, the significant swine pathogen, causes infections characterized by growth and development disorders, skin lesions, and respiratory distress. PCV3 has been circulating worldwide and can be associated with various clinical signs and disease developments. Wild boars are the main reservoir of these pathogens in wildlife and can create an alarming threat to pig farming. In Russia, three PCV2 genotypes (PCV2a, PCV2b, and PCV2d) were identified in pig farms. Additionally, PCV3 was observed in pig herds during the monitoring studies in the country. However, data considering the circulation of PCVs in herds of wild boars in Russia is scant. For this purpose, we performed PCR assays of the samples from 30 wild boars hunted in the Moscow Region of Russia in 2021–2023. The ratios of wild boars positive for PCV2, PCV3, or coinfected were 50, 10, and 13.3%, respectively. Additionally, we sequenced 15 PCV2 and four PCV3 complete genomes and conducted phylogenetic analysis, which divided PCV2 isolates into two groups: PCV2d and PCV2b. The study showed a high infection rate of PCV2 among wild boars, with PCV2d dominance. Simultaneously, PCV3 also circulates among wild boars. The obtained results can provide a basis for the development of preventive measures to support infection transmission risks between farm and wild animals. [ABSTRACT FROM AUTHOR]

Introduction: The escalating occurrence of infectious disease outbreaks in humans and animals necessitates innovative, effective, and integrated research to better comprehend their transmission and dynamics. Viral infection in livestock has led to profound economic losses globally. Pneumonia is the prevalent cause of death in sheep. However, very few studies exist regarding virus-related pathogens in sheep. Metagenomics sequencing technologies in livestock research hold significant potential to elucidate these contingencies and enhance our understanding. Methods: Therefore, this study aims to characterize respiratory viromes in paired nasal swabs from Inner Mongolian feedlot sheep in China using metaviromic sequencing. Through deep sequencing, de novo assembly, and similarity searches using translated protein sequences, several previously uncharacterized and known viruses were identified in this study. Results: Among these discoveries, a novel Bovine Rhinitis B Virus (BRBV) (BRBVsheep) strain was serendipitously detected in the nasal swabs of domestic sheep (Ovis aries). To facilitate further molecular epidemiological studies, the entire genome of BRBV-sheep was also determined. Owing to the unique sequence characteristics and phylogenetic position of BRBV-sheep, genetically distinct lineages of BRBV in sheep may exist. A TaqMan-based qRT-PCR assay targeting the 3D polymerase gene was developed and used to screen 592 clinical sheep specimens. The results showed that 44.59% of the samples (264/592) were positive. These findings suggest that BRBV sheep are widespread among Inner Mongolian herds. Conclusion: This discovery marks the initial identification of BRBV in sheep within Inner Mongolia, China. These findings contribute to our understanding of the epidemiology and genetic evolution of BRBV. Recognizing the presence of BRBV in sheep informs strategies for disease management and surveillance and the potential development of targeted interventions to control its spread. [ABSTRACT FROM AUTHOR]

Dickeya dadantii is a common pathogen of bacterial soft rot on a wide range of plants, including several crops. In this study, we present the complete genome sequence of the D. dadantii type strain DSM18020T. The genome was assembled using PacBio technology, resulting in a 4,997,541 bp circular chromosome with a G+C content of 56.5%. Our sequence analyses predicted 4277 protein-encoding genes, including several associated with known bacterial virulence factors and secondary metabolites. Comparative genomics analysis between Dickeya revealed that the category of 'metabolism' is the most important in both the core and accessory genomes, while the category of 'information storage and processing' is the most dominant in unique genomes. These findings will not only help us to understand the pathogenic mechanisms of D. dadantii DSM18020T, but also provide us with useful information for new control strategies against this phytopathogen. [ABSTRACT FROM AUTHOR]

IntroductionMore than 80 viral diseases of grapevines have been reported worldwide. The infectious degeneration disease complex causes growth reduction, stunting, shortening of internodes, bushy growth and decline in susceptible vines. Grapevine fanleaf viruses (GFLV), arabis mosaic virus (ArMV), grapevine deformation virus (GDefV), tomato ring spot virus (ToRSV) and tobacco ring spot virus (TRSV), which belong to the genus Nepovirus, are known to cause infectious degeneration. The genus Nepovirus has been divided into three subgroups A, B and C based on genome lenth, genomic organization, serological relationships, proteinase cleavage sites and the phylogenetic relationship of their coat protein (CP) gene. GDefV with the new name Nepovirus deformationis belongs to subgroup A of the genus Nepovirus and is closely related to ArMV and GFLV. Prevalence of GFLV in the vineyards of Iran raises the possibility that a mixed infection of GFLV and GDefV is also present in these areas. In Khorasan-Razavi Province, Iran's third-largest grape production center, research has addressed grape-affecting viruses with potential economic consequences, including reduced yield and product quality. Studying the distribution and genetic diversity of GDefV and other grape viruses is crucial for developing effective management and prevention strategies in the region's vineyards. In this study, GDefV was identified in the vineyards of Khorasan-Razavi province in Northeastern Iran and its complete genome was sequenced. In addition, the phylogenetic relationship of these isolates to other GDefV isolates deposited in GenBank was analyzed. Materials and MethodsSamples from three grapevines were collected from a vineyard in Kashmer, Khorasan-Razavi Province; total RNA was extracted from the petiole of young leaves using the CTAB-PVPP method., GDefV and mixed infections of GDefV/GFLV were detected by PCR using specific primers. The PCR products were electrophoresed in a 1% agarose gel and sequenced. The miRNAs were extracted from grapevine petiole tissue using the modified CTAB method, and small RNA libraries were prepared using the TruSeq Small RNA Sample Prep Kit and sequenced on the Illumina Novaseq 6000 platform. After trimming the reads, contigs were generated using k-mer 15 in Velvet assembler 0.7.31. Contigs were verified using BLASTn and BLASTx in NCBI, and reconstruction of the GDefV genome from the NGS reads was faformed using CLC Genomics Workbench (CLC Bio) software. Phylogenetic trees were generated in MEGA7 using the maximum likelihood (ML) method with 1000 replicates in the bootstrap test. The occurrence of possible recombinations in the GDefV genome was analyzed using the RDP v.6 package. Results and DiscussionThe samples showed leaf deformation, shortening of internodes, bushy growth, stunting and decline, but these symptoms are probably related to GFLV, and GDefV only causes leaf deformation in the infected vine. The PCR amplification and sequencing of a segment of the coat protein gene revealed a co-infection of GDefV and GFLV in the samples. GFLV is widely distributed in Iranian vineyards, GDefV has also been previously reported in the vineyards of northwestern Iran, but this is the first report of GDefV in the vineyards of Khorasan-Razavi Province.After refining the reads, approximately 15 million reads (92-99.7% of the original reads) remained for further analysis. The read sequences of each library were deposited in the Sequence Read Archive (SRA) under the accession number SAMN33747579-81. Three Iranian GDefV isolates obtained from three miRNA libraries were deposited in GenBank (accession numbers **-**).RNA1 and RNA2 of the three Iranian GDefV isolates had a lenth of 7386 and 3753 nucleotides, respectively. The RNA1 in GDefV had an open reading frame (ORF) of 6852 nucleotides in lenth, which started with the start codon AUG at position 288 and ended with the stop codons UAA or UAG at position 7142. The 5' end of the genome had 287 nucleotides long and contained two repeating sequences of 15 nucleotides that formed stem-loop structures. The lenth of the non-coding region at the 3' end had 244 nucleotides. Translation of RNA1 of the GDefV genome produces a polyprotein (p1) of 2285 amino acids (approximately 252 kda). The polyprotein p1 comprises the cofactor proteins proteinase, helicase, VPg, proteinase and polymerase with approximate weights of 45, 88, 3, 25 and 91 kDa, respectively. RNA2 also has one ORF, located between nucleotides 237 and 3560. This open reading frame also produces a 122 kDa polyprotein (P2) with 1108 amino acids. The second fragment of the GDeFV genome had 236 nucleotides as a 5'-noncoding region with three repeating sequences of 15 nucleotides that produced stem-loop structures with a 3-nucleotide loop and a 6bp stem. The 3'-noncoding region was also 193 nucleotides long. Polyprotein P2 comprised protein 2A, movement protein (MP) and coat protein (CP). The GDefV polyproteins had cysteine/alanine (C/A) and arginine/glycine (R/G) cleavage sites similar to those of the GFLV polyprotein. Comparison of the RNA1 sequences from three Iranian GDefV isolates with other GDefV isolates available in GenBank showed that the Iranian isolates had 88.1-92.2 % nucleotide identity with each other and 90.3-93.9 % with GenBank isolates at the nucleotide level. At the amino acid level, the Iranian isolates were 86.6-91.6 % identity with each other and 88.9-92.7 % with GenBank isolates. For RNA2, the Iranian isolates showed 89.4-92 % similar to each other and 89.6-94.2 % similar to GenBank isolates at the nucleotide level. The amino acid similarity between the Iranian GDefV isolates was 85-88.6 %. In the phylogenetic tree based on the nucleotide and amino acid sequences of RNA1 and RNA2 of the GDefV genome, the Iranian isolates of this study were clustered in a distinct clade than other GDefV isolates from Turkey (HE613269 and NC017939).GDefV was reported in 2003 and no further information is available on its distribution in vineyards around the world. GDefV has already been reported from Turkey and Iran, but the complete genome sequence of the Iranian GDefV isolate is being reported for the first time. Further studies on the population diversity of GDefV isolates in different regions of Iran are required to gain more insight into the mechanisms affecting the dynamics of GDefV populations. [ABSTRACT FROM AUTHOR]

Klebsiella pneumoniae Ch1-39 was isolated from chili powder elaborated at San Luis Potosí, México. This microorganism can be found in diverse ecological niches as water, soil, air, plants and hospital setting, it is considered as a relevant opportunistic pathogen causing several diseases and showing increasingly multi-resistance to antibiotics. The genome was sequenced on the Illumina NovaSeq platform and bioinformatic analyses were made at the Bacterial and Viral Bioinformatics Resource Center (BV-BRC). The genome consisted of 72 contigs with a total size of 5,410,125 bp, 5,361 protein coding sequences (CDS), a total of 6 rRNA and 76 tRNA with an average G + C content of 57.22 %. The genome data was deposited at National Center for Biotechnology Information (NCBI) under accession number Bioproject ID PRJNA1062060, Bio Sample ID SAMN40269967. The genome accession number was JBAWUH000000000.

Strain WZ-12, isolated from the rhizosphere soil of star anise trees, exhibits significant antifungal efficacy against a variety of fungi, including Colletotrichum horii, the pathogenic fungus responsible for anthracnose in star anise. Based on 16S rRNA gene sequencing and phylogenetic analysis, the strain was identified as a member of the Streptomyces genus. The genome of WZ-12 comprises a linear chromosome of 9,169,400 bp and a plasmid of 378,325 bp, possessing a G + C content of 71.12%. Of note, the genome encompasses 8,172 protein-coding genes, 21 rRNA genes, 74 tRNA genes, and 89 other noncoding RNA genes. The COG, Swiss-Prot, GO, KEGG, and NR databases annotated up to 79.17, 50.50, 13.17, 37.51, and 90.43% of potential genes, respectively. Remarkably, the genome hosts 175 antimicrobial resistance genes and 33 secondary metabolite gene clusters that likely contribute to the biosynthesis of diverse bioactive compounds such as antibiotics and agents with antitumor, antioxidant, immunosuppressive, anti-inflammatory, or stress tolerance properties. These findings highlight the potential of WZ-12 as a promising source of novel bioactive molecules and suggest its significant value as a resource for developing innovative antimicrobial agents. [Figure: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Abstract Objectives The rising of antibiotic resistance has sparked a renewed interest in mycobacteriophage as alternative therapeutic strategies against mycobacterial infections. So far, the vast majority of mycobacteriophages have been isolated using the model species Mycobacterium smegmatis, implying an overwhelming majority of mycobacteriophages in the environment remain uncultured, unclassified, and their specific hosts and infection strategies are still unknown. This study was undertaken to isolate and characterize novel mycobacteriophages targeting Mycobacterium septicum. Data description Here a novel mycobacteriophage WXIN against M. septicum was isolated from soil samples in Wuhan, China. Whole genome analysis indicates that the phage genome consists of 115,158 bp with a GC content of 61.9%. Of the 260 putative open reading frames, 46 may be associated with phage packaging, structure, lysis, lysogeny, genome modification/replication, and other functional roles. The limited genome-wide similarity, along with phylogenetic trees constructed based on viral proteome and orthologous genes show that phage WXIN represents a novel cluster distantly related to cluster J mycobacteriophages (genus Omegavirus). Overall, these results provide novel insights into the genomic properties of mycobacteriophages, highlighting the great genetic diversity of mycobacteriophages in relation to their hosts.

Abstract Objectives Knoxia roxburghii is a member of the madder (Rubiaceae) family. This plant is cultivated in different areas of China and recognized for its medicinal properties, which leads to its use in traditional Chinese medicine. The incidence of root rot was 10–15%. In June 2023, the causal agent of root rot on K. roxburghii was identified as Fusarium oxysporum. To the best of our knowledge, this is the first report of the complete genome of F. oxysporum strain ByF01 that is the causal agent of root rot of K. roxburghii in China. The results will provide effective resources for pathogenesis on K. roxburghii and the prevention and control of root rot on this host in the future. Data description To understand the molecular mechanisms used by F. oxysporum to cause root rot on K. roxburghii, strain ByF01 was isolated from diseased roots and identified by morphological and molecular methods. The complete genome of strain ByF01 was then sequenced using a combination of the PacBio Sequel IIe and Illumina sequencing platforms. We obtained 54,431,725 bp of nucleotides, 47.46% GC content, and 16,705 coding sequences.

Abstract Endophytes have been shown to promote plant growth and health. In the present study, a Bacillus velezensis CH1 (CH1) strain was isolated and identified from high-quality oats, which was capable of producing indole-3-acetic acid (IAA) and strong biofilms, and capabilities in the nitrogen-fixing and iron carriers. CH1 has a 3920 kb chromosome with 47.3% GC content and 3776 code genes. Compared genome analysis showed that the largest proportion of the COG database was metabolism-related (44.79%), and 1135 out of 1508 genes were associated with the function “biosynthesis, transport, and catabolism of secondary metabolites.” Furthermore, thirteen gene clusters had been identified in CH1, which were responsible for the synthesis of fifteen secondary metabolites that exhibit antifungal and antibacterial properties. Additionally, the strain harbors genes involved in plant growth promotion, such as seven putative genes for IAA production, spermidine and polyamine synthase genes, along with multiple membrane-associated genes. The enrichment of these functions was strong evidence of the antimicrobial properties of strain CH1, which has the potential to be a biofertilizer for promoting oat growth and disease resistance.

ABSTRACT: Objectives: Burkholderia dolosa is a clinically important opportunistic pathogen in inpatients. Here we characterised an extensively drug-resistant and hypervirulent B. dolosa isolate from a patient hospitalised for stroke. Methods: Resistance to 41 antibiotics was tested with the agar disc diffusion, minimum inhibitory concentration, or broth microdilution method. The complete genome was assembled using short-reads and long-reads and the hybrid de novo assembly method. Allelic profiles obtained by multilocus sequence typing were analysed using the PubMLST database. Antibiotic-resistance and virulence genes were predicted in silico using public databases and the ‘baargin’ workflow. B. dolosa N149 phylogenetic relationships with all available B. dolosa strains and Burkholderia cepacia complex strains were analysed using the pangenome obtained with Roary. Results: B. dolosa N149 displayed extensive resistance to 31 antibiotics and intermediate resistance to 4 antibiotics. The complete genome included three circular chromosomes (6 338 630 bp in total) and one plasmid (167 591 bp). Genotypic analysis revealed various gene clusters (acr, amr, amp, emr, ade, bla and tet) associated with resistance to 35 antibiotic classes. The major intrinsic resistance mechanisms were multidrug efflux pump alterations, inactivation and reduced permeability of targeted antibiotics. Moreover, 91 virulence genes (encoding proteins involved in adherence, formation of capsule, biofilm and colony, motility, phagocytosis inhibition, secretion systems, protease secretion, transmission and quorum sensing) were identified. B. dolosa N149 was assigned to a novel sequence type (ST2237) and formed a mono-phylogenetic clade separated from other B. dolosa strains. Conclusions: This study provided insights into the antimicrobial resistance and virulence mechanisms of B. dolosa.

Abstract Bacterial blight of rice is a devastating disease caused by the gram-negative bacteria Xanthomonas oryzae pv. oryzae (Xoo). Chinese Xoo strain pathotypes IV, V, and IX are the major virulent Xoo strain types in South China sequentially from the 1990s to the present. Here, we report the isolation of GD0201 and GD0202, which belong to pathotypes IV and IX, respectively, and the complete genome sequence and transcriptomic analysis of GD0201 (IV), GD1358 (V), and GD0202 (IX). We found that resistance genes xa5, Xa23, and Xa27 confer strong resistance to all three Xoo strains, indicating that they are currently good choices for resistance rice breeding. The genome analysis reveals fewer TAL and non-TAL effector coding genes in GD0202 than in the other two strains, potentially contributing to its strong virulence. Transcriptomic analysis of ZH11 inoculated with the three Xoo strains strongly suggests that three Xoo strains for better infection repress the ethylene response factor (ERF) gene family members. Furthermore, weighted gene co-expression network analysis (WGCNA) and protein-protein interaction (PPI) analysis revealed 14 hub genes potentially associated with rice response to the three Xoo strains. The expression of several hub genes was validated to be induced by all three Xoo strains, suggesting its role in bacterial blight disease response to Xoo strains. Genomic analysis of the Xoo strains belonging to pathotypes IV, V, and IX, identification of effectors and genes related to Xoo virulence in rice plants will provide insights into understanding the molecular mechanism underlying rice-Xoo interaction and the gene expression pattern in response to Xoo infection.

Abstract Background Pseudomonas syringae pv. actinidiae (Psa) is an important bacterial plant pathogen that causes severe damage to the kiwifruit industry worldwide. Three Psa strains were recently obtained from different kiwifruit orchards in Anhui Province, China. The present study mainly focused on the variations in virulence and genome characteristics of these strains based on the pathogenicity assays and comparative genomic analyses. Results Three strains were identified as biovar 3 (Psa3), along with strain QSY6 showing higher virulence than JZY2 and YXH1 in pathogenicity assays. The whole genome assembly revealed that each of the three strains had a circular chromosome and a complete plasmid. The chromosome sizes ranged from 6.5 to 6.6 Mb with a GC content of approximately 58.39 to 58.46%, and a predicted number of protein-coding sequences ranging from 5,884 to 6,019. The three strains clustered tightly with 8 Psa3 reference strains in terms of average nucleotide identity (ANI), whole-genome-based phylogenetic analysis, and pangenome analysis, while they were evolutionarily distinct from other biovars (Psa1 and Psa5). Variations were observed in the repertoire of effectors of the type III secretion system among all 15 strains. Moreover, synteny analysis of the three sequenced strains revealed eight genomic regions containing 308 genes exclusively present in the highly virulent strain QSY6. Further investigation of these genes showed that 16 virulence-related genes highlight several key factors, such as effector delivery systems (type III secretion systems) and adherence (type IV pilus), which might be crucial for the virulence of QSY6. Conclusion Three Psa strains were identified and showed variant virulence in kiwifruit plant. Complete genome sequences and comparative genomic analyses further provided a theoretical basis for the potential pathogenic factors responsible for kiwifruit bacterial canker.

Objectives: The rising of antibiotic resistance has sparked a renewed interest in mycobacteriophage as alternative therapeutic strategies against mycobacterial infections. So far, the vast majority of mycobacteriophages have been isolated using the model species Mycobacterium smegmatis, implying an overwhelming majority of mycobacteriophages in the environment remain uncultured, unclassified, and their specific hosts and infection strategies are still unknown. This study was undertaken to isolate and characterize novel mycobacteriophages targeting Mycobacterium septicum. Data description: Here a novel mycobacteriophage WXIN against M. septicum was isolated from soil samples in Wuhan, China. Whole genome analysis indicates that the phage genome consists of 115,158 bp with a GC content of 61.9%. Of the 260 putative open reading frames, 46 may be associated with phage packaging, structure, lysis, lysogeny, genome modification/replication, and other functional roles. The limited genome-wide similarity, along with phylogenetic trees constructed based on viral proteome and orthologous genes show that phage WXIN represents a novel cluster distantly related to cluster J mycobacteriophages (genus Omegavirus). Overall, these results provide novel insights into the genomic properties of mycobacteriophages, highlighting the great genetic diversity of mycobacteriophages in relation to their hosts. [ABSTRACT FROM AUTHOR]

Bovine torovirus (BToV) is an enteric pathogen that may cause diarrhea in calves and adult cattle, which could result in economic losses due to weight loss and decreased milk production. This study aimed to report the presence, the genetic characterization and the evolution of BToV in calves in Uruguay. BToV was detected in 7.9% (22/278) of fecal samples, being identified in dairy (9.2%, 22/239) but not beef (0.0%, 0/39) calves. BToV was detected in both diarrheic (14%, 6/43) and non-diarrheic (13.2%, 5/38) dairy calves. In addition, BToV was detected in the intestinal contents of 14.9% (7/47) of naturally deceased dairy calves. A complete genome (28,446 nucleotides) was obtained, which was the second outside Asia and the first in Latin America. In addition, partial S gene sequences were obtained to perform evolutionary analyses. Nucleotide and amino acid substitutions within and between outbreaks/farms were observed, alerting the continuous evolution of the virus. Through Bayesian analysis using BEAST, a recent origin (mid-60s) of BToV, possibly in Asia, was estimated, with two introductions into Uruguay from Asia and Europe in 2004 and 2013, respectively. The estimated evolutionary rate was 1.80 × 10−3 substitutions/site/year. Our findings emphasize the importance of continued surveillance and genetic characterization for the effective management and understanding of BToV's global epidemiology and evolution. [ABSTRACT FROM AUTHOR]

Plasmodiophora brassicae (Woronin, 1877), a biotrophic, obligate parasite, is the causal agent of clubroot disease in brassicas. The clubroot pathogen has been reported in more than 80 countries worldwide, causing economic losses of hundreds of millions every year. Despite its widespread impact, very little is known about the molecular strategies it employs to induce the characteristic clubs in the roots of susceptible hosts during infection, nor about the mechanisms it uses to overcome genetic resistance. Here, we provide the first telomere-to-telomere complete genome of P. brassicae. We generated ∼27 Gb of Illumina, Oxford Nanopore, and PacBio HiFi data from resting spores of strain Pb3A and produced a 25.3 Mb assembly comprising 20 chromosomes, with an N50 of 1.37 Mb. The BUSCO score, the highest reported for any member of the group Rhizaria (Eukaryota: 88.2%), highlights the limitations within the Eukaryota database for members of this lineage. Using available transcriptomic data and protein evidence, we annotated the Pb3A genome, identifying 10,521 protein-coding gene models. This high-quality, complete genome of P. brassicae will serve as a crucial resource for the plant pathology community to advance the much-needed understanding of the evolution of the clubroot pathogen. [ABSTRACT FROM AUTHOR]

Salmonella diarizonae (IIIb) is frequently isolated from reptiles and less frequently from birds and mammals. However, its isolation from invasive human infections has not been widely reported. Migratory mallard ducks are excellent bioindicators of pathogen presence and pathogen antibiotic resistance (AMR). We present the first isolation from a mallard duck in central Europe of the antibiotic-resistant Salmonella enterica subsp. diarizonae with the unique antigenic pattern 58:r:z53 and report its whole-genome sequencing, serosequencing, and genotyping, which enabled the prediction of its pathogenicity and comparison with phenotypic AMR. The isolated strain was highly similar to S. diarizonae isolated from humans and food. Twenty-four AMR genes were detected, including those encoding aminoglycoside, fluoroquinolone, macrolide, carbapenem, tetracycline, cephalosporin, nitroimidazole, peptide antibiotic, and disinfecting agent/antiseptic resistance. Six Salmonella pathogenicity islands were found (SPI-1, SPI-2, SPI-3, SPI-5, SPI-9, and SPI-13). An iron transport system was detected in SPI-1 centisome C63PI. Plasmid profile analyses showed three to be present. Sequence mutations in the invA and invF genes were noted, which truncated and elongated the proteins, respectively. The strain also harbored genes encoding type-III secretion-system effector proteins and many virulence factors found in S. diarizonae associated with human infections. This study aims to elucidate the AMR and virulence genes in S. enterica subsp. diarizonae that may most seriously threaten human health. [ABSTRACT FROM AUTHOR]

Bovine herpesvirus 4 (BoHV-4) is an indigenous virus in cattle prevalent mainly in North and South American countries and European countries, but the genomic sequences and genetic characteristics of Japanese strains have not been reported. BoHV-4 is suspected, but not proven, to be associated with various diseases. In the present study, we isolated BoHV-4 from a 10-month-old Japanese Black calf with respiratory symptoms in Japan. To identify the genetic characteristics of the isolate named strain SG20, complete genome sequencing was performed using a combination of next-generation and Sanger sequencing technologies. The complete long unique coding region (LUR) of SG20 was found to comprise 108,819 nucleotides with 41.4% GC content and contain at least 78 open reading frames. It shares 83.4 to 99.3% overall nucleotide identity with six BoHV-4 strains available in the database. The deduced amino acid sequence alignment revealed that SG20 contains genotype 1-specific features of BoHV-4, such as amino acid substitutions and insertions within the glycoprotein B region. Phylogenetic analyzes based on the nucleotide sequences of ORF20 indicated that the virus belonged to genotype 1 (Movar 33/63-like group). The strain was also analyzed using the complete LUR and placed in the same clade as a strain recently isolated from China, but it was distinct from American and European BoHV-4 strains of genotype 1. Although further genomic and epidemiologic information is needed, our results help elucidate the molecular epidemiology of BoHV-4 and provide a foundation for future studies. [ABSTRACT FROM AUTHOR]

Introduction More than 80 viral diseases of grapevines have been reported worldwide. The infectious degeneration disease complex causes growth reduction, stunting, shortening of internodes, bushy growth and decline in susceptible vines. Grapevine fanleaf viruses (GFLV), arabis mosaic virus (ArMV), grapevine deformation virus (GDefV), tomato ring spot virus (ToRSV) and tobacco ring spot virus (TRSV), which belong to the genus Nepovirus, are known to cause infectious degeneration. The genus Nepovirus has been divided into three subgroups A, B and C based on genome length, genomic organization, serological relationships, proteinase cleavage sites and the phylogenetic relationship of their coat protein (CP) gene. GDefV with the new name Nepovirus deformationis belongs to subgroup A of the genus Nepovirus and is closely related to ArMV and GFLV. Prevalence of GFLV in the vineyards of Iran raises the possibility that a mixed infection of GFLV and GDefV is also present in these areas. In Khorasan-Razavi Province, Iran's third-largest grape production center, research has addressed grape-affecting viruses with potential economic consequences, including reduced yield and product quality. Studying the distribution and genetic diversity of GDefV and other grape viruses is crucial for developing effective management and prevention strategies in the region's vineyards. In this study, GDefV was identified in the vineyards of Khorasan-Razavi province in Northeastern Iran and its complete genome was sequenced. In addition, the phylogenetic relationship of these isolates to other GDefV isolates deposited in GenBank was analyzed. Materials and Methods Samples from three grapevines were collected from a vineyard in Kashmer, Khorasan-Razavi Province; total RNA was extracted from the petiole of young leaves using the CTAB-PVPP method., GDefV and mixed infections of GDefV/GFLV were detected by PCR using specific primers. The PCR products were electrophoresed in a 1% agarose gel and sequenced. The miRNAs were extracted from grapevine petiole tissue using the modified CTAB method, and small RNA libraries were prepared using the TruSeq Small RNA Sample Prep Kit and sequenced on the Illumina Novaseq 6000 platform. After trimming the reads, contigs were generated using k-mer 15 in Velvet assembler 0.7.31. Contigs were verified using BLASTn and BLASTx in NCBI, and reconstruction of the GDefV genome from the NGS reads was performed using CLC Genomics Workbench (CLC Bio) software. Phylogenetic trees were generated in MEGA7 using the maximum likelihood (ML) method with 1000 replicates in the bootstrap test. The occurrence of possible recombinations in the GDefV genome was analyzed using the RDP v.6 package. Results and Discussion The samples showed leaf deformation, shortening of internodes, bushy growth, stunting and decline, but these symptoms are probably related to GFLV, and GDefV only causes leaf deformation in the infected vine. The PCR amplification and sequencing of a segment of the coat protein gene revealed a co-infection of GDefV and GFLV in the samples. GFLV is widely distributed in Iranian vineyards, GDefV has also been previously reported in the vineyards of northwestern Iran, but this is the first report of GDefV in the vineyards of Khorasan-Razavi Province. After refining the reads, approximately 15 million reads (92-99.7% of the original reads) remained for further analysis. The read sequences of each library were deposited in the Sequence Read Archive (SRA) under the accession number SAMN33747579-81. Three Iranian GDefV isolates obtained from three miRNA libraries were deposited in GenBank (accession numbers **-**). RNA1 and RNA2 of the three Iranian GDefV isolates had a length of 7386 and 3753 nucleotides, respectively. The RNA1 in GDefV had an open reading frame (ORF) of 6852 nucleotides in length, which started with the start codon AUG at position 288 and ended with the stop codons UAA or UAG at position 7142. The 5' end of the genome had 287 nucleotides long and contained two repeating sequences of 15 nucleotides that formed stem-loop structures. The length of the non-coding region at the 3' end had 244 nucleotides. Translation of RNA1 of the GDefV genome produces a polyprotein (p1) of 2285 amino acids (approximately 252 kda). The polyprotein p1 comprises the cofactor proteins proteinase, helicase, VPg, proteinase and polymerase with approximate weights of 45, 88, 3, 25 and 91 kDa, respectively. RNA2 also has one ORF, located between nucleotides 237 and 3560. This open reading frame also produces a 122 kDa polyprotein (P2) with 1108 amino acids. The second fragment of the GDeFV genome had 236 nucleotides as a 5'-noncoding region with three repeating sequences of 15 nucleotides that produced stem-loop structures with a 3-nucleotide loop and a 6bp stem. The 3'-noncoding region was also 193 nucleotides long. Polyprotein P2 comprised protein 2A, movement protein (MP) and coat protein (CP). The GDefV polyproteins had cysteine/alanine (C/A) and arginine/glycine (R/G) cleavage sites similar to those of the GFLV polyprotein. Comparison of the RNA1 sequences from three Iranian GDefV isolates with other GDefV isolates available in GenBank showed that the Iranian isolates had 88.1-92.2 % nucleotide identity with each other and 90.3-93.9 % with GenBank isolates at the nucleotide level. At the amino acid level, the Iranian isolates were 86.6-91.6 % identity with each other and 88.9-92.7 % with GenBank isolates. For RNA2, the Iranian isolates showed 89.4-92 % similar to each other and 89.6-94.2 % similar to GenBank isolates at the nucleotide level. The amino acid similarity between the Iranian GDefV isolates was 85-88.6 %. In the phylogenetic tree based on the nucleotide and amino acid sequences of RNA1 and RNA2 of the GDefV genome, the Iranian isolates of this study were clustered in a distinct clade than other GDefV isolates from Turkey (HE613269 and NC017939). GDefV was reported in 2003 and no further information is available on its distribution in vineyards around the world. GDefV has already been reported from Turkey and Iran, but the complete genome sequence of the Iranian GDefV isolate is being reported for the first time. Further studies on the population diversity of GDefV isolates in different regions of Iran are required to gain more insight into the mechanisms affecting the dynamics of GDefV populations. [ABSTRACT FROM AUTHOR]

Bacterial blight of rice is a devastating disease caused by the gram-negative bacteria Xanthomonas oryzae pv. oryzae (Xoo). Chinese Xoo strain pathotypes IV, V, and IX are the major virulent Xoo strain types in South China sequentially from the 1990s to the present. Here, we report the isolation of GD0201 and GD0202, which belong to pathotypes IV and IX, respectively, and the complete genome sequence and transcriptomic analysis of GD0201 (IV), GD1358 (V), and GD0202 (IX). We found that resistance genes xa5, Xa23, and Xa27 confer strong resistance to all three Xoo strains, indicating that they are currently good choices for resistance rice breeding. The genome analysis reveals fewer TAL and non-TAL effector coding genes in GD0202 than in the other two strains, potentially contributing to its strong virulence. Transcriptomic analysis of ZH11 inoculated with the three Xoo strains strongly suggests that three Xoo strains for better infection repress the ethylene response factor (ERF) gene family members. Furthermore, weighted gene co-expression network analysis (WGCNA) and protein-protein interaction (PPI) analysis revealed 14 hub genes potentially associated with rice response to the three Xoo strains. The expression of several hub genes was validated to be induced by all three Xoo strains, suggesting its role in bacterial blight disease response to Xoo strains. Genomic analysis of the Xoo strains belonging to pathotypes IV, V, and IX, identification of effectors and genes related to Xoo virulence in rice plants will provide insights into understanding the molecular mechanism underlying rice-Xoo interaction and the gene expression pattern in response to Xoo infection. [ABSTRACT FROM AUTHOR]

Fermented eggplant is a traditional fermented food, however lactic acid bacteria capable of producing exopolysaccharide (EPS) have not yet been exploited. The present study focused on the production and protective effects against oxidative stress of an EPS produced by Lacticaseibacillus paracasei NC4 (NC4-EPS), in addition to deciphering its genomic features and EPS biosynthesis pathway. Among 54 isolates tested, strain NC4 showed the highest EPS yield and antioxidant activity. The maximum EPS production (2.04 ± 0.11 g/L) was achieved by culturing in MRS medium containing 60 g/L sucrose at 37 °C for 48 h. Under 2 mM H2O2 stress, the survival of a yeast model Saccharomyces cerevisiae treated with 0.4 mg/mL NC4-EPS was 2.4-fold better than non-treated cells, which was in agreement with the catalase and superoxide dismutase activities measured from cell lysates. The complete genome of NC4 composed of a circular chromosome of 2,888,896 bp and 3 circular plasmids. The NC4 genome comprises more genes with annotated function in nitrogen metabolism, phosphorus metabolism, cell division and cell cycle, and iron acquisition and metabolism as compared to other reported L. paracasei. Of note, the eps gene cluster is not conserved across L. paracasei. Pathways of sugar metabolism for EPS biosynthesis were proposed for the first time, in which gdp pathway only present in few plant-derived bacteria was identified. These findings shed new light on the cell-protective activity and biosynthesis of EPS produced by L. paracasei, paving the way for future efforts to enhance yield and tailor-made EPS production for food and pharmaceutical industries. [ABSTRACT FROM AUTHOR]

Background: Pseudomonas syringae pv. actinidiae (Psa) is an important bacterial plant pathogen that causes severe damage to the kiwifruit industry worldwide. Three Psa strains were recently obtained from different kiwifruit orchards in Anhui Province, China. The present study mainly focused on the variations in virulence and genome characteristics of these strains based on the pathogenicity assays and comparative genomic analyses. Results: Three strains were identified as biovar 3 (Psa3), along with strain QSY6 showing higher virulence than JZY2 and YXH1 in pathogenicity assays. The whole genome assembly revealed that each of the three strains had a circular chromosome and a complete plasmid. The chromosome sizes ranged from 6.5 to 6.6 Mb with a GC content of approximately 58.39 to 58.46%, and a predicted number of protein-coding sequences ranging from 5,884 to 6,019. The three strains clustered tightly with 8 Psa3 reference strains in terms of average nucleotide identity (ANI), whole-genome-based phylogenetic analysis, and pangenome analysis, while they were evolutionarily distinct from other biovars (Psa1 and Psa5). Variations were observed in the repertoire of effectors of the type III secretion system among all 15 strains. Moreover, synteny analysis of the three sequenced strains revealed eight genomic regions containing 308 genes exclusively present in the highly virulent strain QSY6. Further investigation of these genes showed that 16 virulence-related genes highlight several key factors, such as effector delivery systems (type III secretion systems) and adherence (type IV pilus), which might be crucial for the virulence of QSY6. Conclusion: Three Psa strains were identified and showed variant virulence in kiwifruit plant. Complete genome sequences and comparative genomic analyses further provided a theoretical basis for the potential pathogenic factors responsible for kiwifruit bacterial canker. [ABSTRACT FROM AUTHOR]

A Gram-negative, facultatively anaerobic, short rod-shaped bacterium, designated as strain HZ0627T, was isolated from the appendiceal pus of a patient with appendicitis in Yongzhou, Hunan, China. This strain was subjected to comprehensive phenotypic, phylogenetic, and genomic analyses using polyphasic taxonomic methods. Phylogenetic analysis of the 16S rRNA gene sequence revealed that this strain belonged to the genus Proteus and the family Morganellaceae, whereas that based on the rpoB gene sequence and phylogenomic analysis demonstrated that this strain was distinctly separated from other type strains of Proteus species. Moreover, whole-genome-based analyses, including in silico DNA–DNA hybridization (isDDH) and average nucleotide identity (ANI), revealed that strain HZ0627T had much lower isDDH rates (24.5–55.6%) and ANI (82.04–93.90%) than those of the thresholds (i.e., 70% and 95%, respectively) for species delineation, when compared to the type strains of other Proteus species. The cellular fatty acid profile of strain HZ0627T was dominated by C16:0 (34.5%), cyclo C17:0 (25.8%), C14:0 (12.6%), C16:1 iso I/14:0 3-OH (7.7%), C18:1ω7c/18:1ω6c (6.5%), and C16:1ω7c/16:1ω6c (4.9%), which clearly differentiated it from the documented type strains of Proteus species. In addition, several specific physiological traits, including optimal growth temperature, tolerance to sodium chloride, and carbon source utilization, differed from those of other Proteus species. Therefore, we propose the name Proteus appendicitidis sp. nov. for strain HZ0627T (= CCTCC AB 2022380T = KCTC 92986T), which represents the type strain of this novel Proteus species. [ABSTRACT FROM AUTHOR]

ABSTRACT: Enteropathies are a serious concern in racing pigeons as they significantly impair performance in races and their training, and viruses are suspected to be one of the main factors. Astroviruses are well-known to be responsible for causing enteric disease in humans and various other animals including birds, although their prevalence and pathogenicity in pigeons is poorly understood. In this study, we investigated 2 groups of young racing pigeons (sick—study group and healthy—control group) to assess the correlation between the number of astrovirus genome copies in cloacal swabs and the occurrence of enteropathy. To determine this, we developed a novel TaqMan quantitative PCR (qPCR) and digital droplet PCR (ddPCR) methods for astrovirus detection and absolute quantitative analysis. We also performed high-throughput sequencing to obtain the complete genome sequences and establish the genetic similarity of the obtained strains to known astroviruses of poultry and other avian species. Two new complete genome sequences of pigeon astroviruses in the Avastrovirus genus were identified, representing 2 new species. These were found most closely related to astroviruses identified in Columbidae species and chickens. They share an average of 75.8% genome-wide pairwise identity and 57.6% and 64.6% capsid protein sequence identity with other unclassified columbid avastrovirus sequences in GenBank. Although the difference in prevalence of astrovirus in the study and control group was found statistically insignificant, there was a significant difference between the number of genome copies in positive samples from both groups. These unambiguous results leave the role of astroviruses as enteropathogenic factors in pigeons still undetermined.

Viruses are among the main pathogens causing diarrhoea in calves. The current study found that bovine norovirus (BNoV) is one of the principal viruses causing diarrhoea in calves in Xinjiang, China.

As an immunosuppressive disease virus, chicken infectious anemia virus (CIAV) mainly infects chickens, causing aplastic anemia and systemic lymphoid tissue atrophy. In recent years, the prevalence of CIAV in the poultry industry globally has caused huge economic losses. In this study, a total of 223 clinical samples, including anal swabs, tissues, blood, and vaccines, were collected from 19 broiler farms or breeding companies in Jiangsu province, with symptoms of significant anemia and immunosuppression during 2020-2022. Among them, 75 samples (75/223, 33.6%) were positive for CIAV in polymerase chain reaction (PCR) test, and 20 CIAV strains were successfully isolated. The phylogenetic trees of the 20 isolates and 42 CIAV strains deposited in GenBank formed four distinct groups (A-D). And the isolates mainly belonged to Group A but with high genetic diversity. Analysis for VP1 indicated that these isolates possess key characteristics of highly pathogenic strains. Meanwhile, VP2 and VP3 were much conserved with much fewer mutations compare to VP1. The above epidemiological study of CIAV provides novel insights into molecular characterization of CIAV and lays the foundation for developing efficient strategies for control of CIAV in China. [ABSTRACT FROM AUTHOR]

In 2015 and 2016, two Barramundi (Lates calcarifer) farms in Singapore reported a disease outbreak characterized by lethargic behavior, pronounced inappetence, generalized skin lesions, erosions of the fins and tail, and ultimately high mortality in their fish. Next-generation sequencing and PCR confirmed presence of a novel virus belonging to the Alloherpesviridae family, Lates calcarifer herpesvirus (LCHV), which was subsequently isolated and cultured. We characterize, for the first time, the complete genome of two cultured LCHV isolates. The genome contains a long unique region of approximately 105,000 bp flanked by terminal repeats of approximately 24,800 bp, of which the first 8.2 kb do not show any similarity to described genomes in the Alloherpesviridae family. The two cultured isolates share 89% nucleotide identity, and their closest relatives are the viruses belonging to the genus Ictalurivirus. Experimental infections using one of the cultured LCHV isolates resulted in identical clinical signs as originally described in the index farm, both in intraperitoneal-injection infected fish and cohabitant fish, with mortality in both groups. Histopathological analysis showed pronounced abnormalities in the gills. Virus culture and PCR analysis confirmed the replication of LCHV in the infected fish, and thus Koch's postulates were fulfilled. [ABSTRACT FROM AUTHOR]

Restricted production of fungal secondary metabolites hinders the ability to conduct comprehensive research and development of novel biopesticides. Okaramine B from Penicillium demonstrates remarkable insecticidal efficacy; however, its biosynthetic yield is low, and its regulatory mechanism remains unknown. The present study found that the yield difference was influenced by fermentation modes in okaramine-producing strains and performed genomic and comparative transcriptome analysis of P. daleae strain NBP-49626, which exhibits significant features. The NBP-49626 genome is 37.4 Mb, and it encodes 10,131 protein-encoding genes. Up to 5097 differentially expressed genes (DEGs) were identified during the submerged and semi-solid fermentation processes. The oka gene cluster, lacking regulatory and transport genes, displayed distinct transcriptional patterns in response to the fermentation modes and yield of Okaramine B. Although transcription trends of most known global regulatory genes are inconsistent with those of oka, this study identified five potential regulatory genes, including two novel Zn(II)2Cys6 transcription factors, Reg2 and Reg19. A significant correlation was also observed between tryptophan metabolism and Okaramine B yields. In addition, several transporter genes were identified as DEGs. These results were confirmed using real-time quantitative PCR. This study provides comprehensive information regarding the regulatory mechanism of Okaramine B biosynthesis in Penicillium and is critical to the further yield improvement for the development of insecticides. [ABSTRACT FROM AUTHOR]

The economically important pale tussock moth Dasychira pudibunda L. (Lepidoptera: Lymantriidae), known as a beech pest in Europe, primarily inflicts damage on beech leaves. In the present study, we aim to reveal the genome characteristics of Dasychira pudibunda nucleopolyhedrovirus (DapuNPV-T1), which was detected for the first time in Turkey and compare it with the reference genome and other baculovirus genomes. The DapuNPV-T1 genome was determined to be a circular, double-stranded DNA molecule with 136,920 bp and a nucleotide distribution of 54.4% G + C. Bioinformatics analysis showed that the DapuNPV-T1 genome contains 163 open reading frames with more than 150 nucleotides. Fifty-four ORFs of unknown function, 6 homologous regions (hrs), 1 AC-rich region, and 3 bro genes (bro-a, bro-b, and bro-c) were determined in the genome sequence. Comparative analysis with other baculovirus strains revealed distinctions in the DapuNPV-T1 genome based on ORF. The gene parity plot and phylogenetic analysis confirmed that DapuNPV-T1 belongs to the alphabaculovirus group Ib. In addition, the DapuNPV-T1 isolate was found to be close to the nucleopolyhedrovirus Poland isolate in Dasychira pudibunda and Orgyia pseudotsugata multiple nucleopolyhedrovirus. With this study, the first genome analysis of DapuNPV from Turkey became the second in the world to enter the literature. Comprehensive information on a wide range of isolates will provide a more detailed overview of baculoviruses and help overcome their shortcomings as biocontrol agents. [ABSTRACT FROM AUTHOR]

Porcine circoviruses (PCVs) are widely distributed in swine herds. PCV2, the significant swine pathogen, causes infections characterized by growth and development disorders, skin lesions, and respiratory distress. PCV3 has been circulating worldwide and can be associated with various clinical signs and disease developments. Wild boars are the main reservoir of these pathogens in wildlife and can create an alarming threat to pig farming. In Russia, three PCV2 genotypes (PCV2a, PCV2b, and PCV2d) were identified in pig farms. Additionally, PCV3 was observed in pig herds during the monitoring studies in the country. However, data considering the circulation of PCVs in herds of wild boars in Russia is scant. For this purpose, we performed PCR assays of the samples from 30 wild boars hunted in the Moscow Region of Russia in 2021–2023. The ratios of wild boars positive for PCV2, PCV3, or coinfected were 50, 10, and 13.3%, respectively. Additionally, we sequenced 15 PCV2 and four PCV3 complete genomes and conducted phylogenetic analysis, which divided PCV2 isolates into two groups: PCV2d and PCV2b. The study showed a high infection rate of PCV2 among wild boars, with PCV2d dominance. Simultaneously, PCV3 also circulates among wild boars. The obtained results can provide a basis for the development of preventive measures to support infection transmission risks between farm and wild animals.