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BMC Research Notes Nov 2014Application of molecular diagnostic methods to the determination of etiology in suspected poxvirus-associated infections of bovines is important both for the diagnosis...
BACKGROUND
Application of molecular diagnostic methods to the determination of etiology in suspected poxvirus-associated infections of bovines is important both for the diagnosis of the individual case and to form a more complete understanding of patterns of strain occurrence and spread. The objective of this study was to identify and characterize bovine-associated zoonotic poxviruses in Bangladesh which are relevant to animal and human health.
FINDINGS
Investigators from the International Center Diarrhoeal Disease Research (icddr,b), the US Centers for Disease Control and Prevention (CDC), and the Bangladesh Department of Livestock Services traveled to three districts in Bangladesh-Siranjganj, Rangpur and Bhola-to collect diagnostic specimens from dairy cattle and buffalo that had symptoms consistent with poxvirus-associated infections. Bovine papular stomatitis virus (BPSV) DNA was obtained from lesion material (teat) and an oral swab collected from an adult cow and calf (respectively) from a dairy production farm in Siranjganj. Pseudocowpox virus (PCPV) DNA signatures were obtained from a scab and oral swab collected from a second dairy cow and her calf from Rangpur.
CONCLUSIONS
We report the first detection of zoonotic poxviruses from Bangladesh and show phylogenetic comparisons between the Bangladesh viruses and reference strains based on analyses of the B2L and J6R loci (vaccinia orthologs). Understanding the range and diversity of different species and strains of parapoxvirus will help to spotlight unusual patterns of occurrence that could signal events of significance to the agricultural and public health sectors.
Topics: Animals; Bangladesh; Cattle; Cattle Diseases; Dairying; Geography; Humans; Parapoxvirus; Phylogeny; Poxviridae Infections; Zoonoses
PubMed: 25410770
DOI: 10.1186/1756-0500-7-816 -
Infection, Genetics and Evolution :... Apr 2023In 2016, the first orf virus, a double-stranded DNA (dsDNA) virus of the genus parapoxvirus, from a muskox was isolated on Victoria Island, Nunavut (NU), Canada. We used...
In 2016, the first orf virus, a double-stranded DNA (dsDNA) virus of the genus parapoxvirus, from a muskox was isolated on Victoria Island, Nunavut (NU), Canada. We used deep sequencing on DNA extracted from orf virus-positive tissues from wild muskoxen from locations on Victoria Island and the adjacent mainland. Orf virus sequence reads derived from four samples were nearly identical. The consensus sequences generated from pooled reads of MxOV comprises of a large contiguous sequence (contig) of 131,759 bp and a smaller right terminal contig of 3552 bp, containing all coding sequences identified as Parapoxvirus. Individual gene comparisons reveal that MxOV shares genetic characteristics with reference strains from both sheep and goat origin. Recombination analysis using Bootscan, MAXCHI, GENECONV, CHIMAERA, SISCAN, and RDP algorithms within the RDP4 software predicted recombination events in two virulence factors, and a large 3000 bp segment of the MxOV genome. Partial B2L nucleotide sequences from strains around the world and other North American isolates were compared to MxOV using MUSCLE alignments and RAxML phylogenetic trees. MxOV was identical to our previously characterized isolate, and shared similarity with orf virus isolated from sheep and goats. The phylogenetic grouping of partial B2L nucleotide sequences did not follow the sample geographic distribution. More full genomes of orf virus, or at least full B2L gene squences, in wildlife are needed especially in North America to better understand the epidemiology of the disease in muskoxen.
Topics: Sheep; Animals; Phylogeny; Canada; Ruminants; Orf virus; Communicable Diseases; Goats; High-Throughput Nucleotide Sequencing
PubMed: 36775047
DOI: 10.1016/j.meegid.2023.105414 -
PLoS Pathogens Oct 2021Viruses have evolved mechanisms to subvert critical cellular signaling pathways that regulate a wide range of cellular functions, including cell differentiation,...
Viruses have evolved mechanisms to subvert critical cellular signaling pathways that regulate a wide range of cellular functions, including cell differentiation, proliferation and chemotaxis, and innate immune responses. Here, we describe a novel ORFV protein, ORFV113, that interacts with the G protein-coupled receptor Lysophosphatidic acid receptor 1 (LPA1). Consistent with its interaction with LPA1, ORFV113 enhances p38 kinase phosphorylation in ORFV infected cells in vitro and in vivo, and in cells transiently expressing ORFV113 or treated with soluble ORFV113. Infection of cells with virus lacking ORFV113 (OV-IA82Δ113) significantly decreased p38 phosphorylation and viral plaque size. Infection of cells with ORFV in the presence of a p38 kinase inhibitor markedly diminished ORFV replication, highlighting importance of p38 signaling during ORFV infection. ORFV113 enhancement of p38 activation was prevented in cells in which LPA1 expression was knocked down and in cells treated with LPA1 inhibitor. Infection of sheep with OV-IA82Δ113 led to a strikingly attenuated disease phenotype, indicating that ORFV113 is a major virulence determinant in the natural host. Notably, ORFV113 represents the first viral protein that modulates p38 signaling via interaction with LPA1 receptor.
Topics: Animals; MAP Kinase Signaling System; Parapoxvirus; Poxviridae Infections; Receptors, Lysophosphatidic Acid; Sheep; Viral Proteins
PubMed: 34614034
DOI: 10.1371/journal.ppat.1009971 -
Frontiers in Microbiology 2019Parapoxvirus of red deer in New Zealand (PVNZ) is a species of the genus that causes pustular dermatitis. We identified a cluster of genes in PVNZ that encode three...
Parapoxvirus of red deer in New Zealand (PVNZ) is a species of the genus that causes pustular dermatitis. We identified a cluster of genes in PVNZ that encode three unique chemokine-binding proteins (CBPs) namely ORF112.0, ORF112.3 and ORF112.6. Chemokines are a large family of molecules that direct cell trafficking to sites of inflammation and through lymphatic organs. The PVNZ-CBPs were analyzed by surface plasmon resonance against a broad spectrum of CXC, CC, XC and CXC chemokines and were found to differ in their specificity and binding affinity. ORF112.0 interacted with chemokines from the CXC, CC and XC classes of chemokines with nM affinities. The ORF112.3 showed a preference for CXC chemokines, while ORF112.6 showed pM affinity binding for CC chemokines. Structural modeling analysis showed alterations in the chemokine binding sites of the CBPs, although the core structure containing two ß-sheets and three α-helices being conserved with the other CBPs. Chemotaxis assays using neutrophils and monocytes revealed inhibitory impact of the CBPs on cell migration. Our results suggest that the PVNZ-CBPs are likely to have evolved through a process of gene duplication and divergence, and may have a role in suppressing inflammation and the anti-viral immune response.
PubMed: 31293551
DOI: 10.3389/fmicb.2019.01421 -
Viruses Jan 2019The (ORFV; ) strain D1701 with an attenuated phenotype and excellent immunogenic capacity is successfully used for the generation of recombinant vaccines against...
The (ORFV; ) strain D1701 with an attenuated phenotype and excellent immunogenic capacity is successfully used for the generation of recombinant vaccines against different viral infections. Adaption for growth in Vero cells was accompanied by additional major genomic changes resulting in ORFV strain variant D1701-V. In this study, restriction enzyme mapping, blot hybridization and DNA sequencing of the deleted region s (A, AT and D) in comparison to the predecessor strain D1701-B revealed the loss of 7 open reading frames (ORF008, ORF101, ORF102, ORF114, ORF115, ORF116, ORF117). The suitability of deletion site D for expression of foreign genes is demonstrated using novel synthetic early promoter eP1 and eP2. Comparison of promoter strength showed that the original promoter Pv as well as promoter eP2 display an up to 11-fold stronger expression than promoter eP1, irrespective of the insertion site. Successful integration and expression of the fluorescent marker genes is demonstrated by gene- and insertion-site specific PCR assays, fluorescence microscopy and flow cytometry. For the first time ORFV recombinants are generated simultaneously expressing transgenes in two different insertion loci. That allows production of polyvalent vaccines containing several antigens against one or different pathogens in a single vectored ORFV vaccine.
Topics: Adaptation, Physiological; Animals; Chlorocebus aethiops; Gene Deletion; Genetic Vectors; Genome, Viral; High-Throughput Nucleotide Sequencing; Orf virus; Recombination, Genetic; Transgenes; Vero Cells
PubMed: 30704093
DOI: 10.3390/v11020127 -
Microbiology and Immunology 2002The possibility of persistent parapoxvirus (PPV) infection was investigated by serologically and genetically using cattle infected with the virus experimentally and...
The possibility of persistent parapoxvirus (PPV) infection was investigated by serologically and genetically using cattle infected with the virus experimentally and naturally. Three cattle were inoculated with the virus subcutaneously at several spots in the lips and abdominal regions. Small papules developed in the inoculated regions, and antibodies to the virus developed and continued persistently. One animal, from which one PPV had been previously isolated, was also subjected to serological and viral detection tests as a naturally infected case. Two of these four cattle were injected with dexamethasone (DM), and one was injected with interferon-gamma (IFN-gamma). The viral genome was rarely detected from the peripheral blood leukocytes in the ordinary condition, but frequently when the animals were injected with IFN-gamma. The viral genome was also detected from the lymph nodes as these PPV infected animals were euthanized. These results indicated that cattle were infected with PPV subclinically and persistently, and the virus was activated in stressed or immunosuppressed animals. The virus would be harbored in the lymphotic tissues of the animals when they show no clinical symptoms.
Topics: Animals; Antibodies, Viral; Blotting, Southern; Cattle; Cattle Diseases; Cells, Cultured; DNA, Viral; Enzyme-Linked Immunosorbent Assay; Immunocompetence; Interferon-gamma; Parapoxvirus; Polymerase Chain Reaction; Poxviridae Infections
PubMed: 12061631
DOI: 10.1111/j.1348-0421.2002.tb02697.x -
BMC Veterinary Research Jan 2019Contagious ecthyma (CE) appears in the countries and regions containing goat and sheep farms, and it is considered a global epidemic. CE not only severely endangers the... (Comparative Study)
Comparative Study
BACKGROUND
Contagious ecthyma (CE) appears in the countries and regions containing goat and sheep farms, and it is considered a global epidemic. CE not only severely endangers the healthy development of the sheep and goat industries but also threatens human health. For viral infectious diseases, fast and effective isolation and culture of the pathogen is critical for CE diagnosis, and for disease prevention and control. Therefore, the sensitivity of bovine Sertoli cells to ORFV was estimate in this study.
RESULTS
The sensitivities of bovine Sertoli cells, primary neonatal bovine testicular cells, and Madin-Darby bovine kidney (MDBK) cell line to ORFV were compared. Our results showed that the isolated bovine Sertoli cells were sensitive to inoculated ORFV, and viral titers were approximately 1 log higher than those in primary neonatal bovine testicular cells and in MDBK cell lines.
CONCLUSION
Appropriately sensitive cells for the highly efficient isolation and culture of the ORFV were obtained. Culture of ORFV using the Sertoli cells showed good consistency and stability and also avoided the risk of other pathogens presenting during viral culture using a primary cell line. In addition, using these passaged bovine Sertoli cells to proliferate ORFV may simplify the CE diagnosis process, thereby reducing detection time and cost. Hence, this test has important practical significance for the diagnosis of CE and the research on the pathogenic mechanism of ORFV.
Topics: Animals; Cattle; Cell Culture Techniques; Cells, Cultured; Ecthyma, Contagious; Male; Orf virus; Sertoli Cells; Virus Replication
PubMed: 30616567
DOI: 10.1186/s12917-018-1760-1 -
The Journal of General Virology Mar 2021Cases of pox-like lesions in horses and donkeys have been associated with poxviruses belonging to different genera of the family . These include the orthopoxviruses...
Cases of pox-like lesions in horses and donkeys have been associated with poxviruses belonging to different genera of the family . These include the orthopoxviruses vaccinia virus (VACV), horsepoxvirus (HPXV) and cowpoxvirus (CPXV), as well as a potentially novel parapoxvirus and molluscum contagiosum virus (MOCV). However, with the exception of VACV, HPXV and CPXV, the genomic characterization of the causative agents remains largely elusive with only single short genome fragments available. Here we present the first full-length genome sequence of an equine molluscum contagiosum-like virus (EMCLV) directly determined from skin biopsies of a horse with generalized papular dermatitis. Histopathological analysis of the lesions revealed severe epidermal hyperplasia with numerous eosinophilic inclusion bodies within keratinocytes. Virions were detected in the lesions in embedded tissue by transmission electron microscopy. The genome sequence determined by next- and third-generation sequencing comprises 166 843 nt with inverted terminal repeats (ITRs) of 3473 nt. Overall, 20 of the predicted 159 ORFs have no equivalents in other poxviruses. Intriguingly, two of these ORFs were identified to encode homologues of mammalian proteins involved in immune signalling pathways, namely (SECTM1) and (IGFLR1), that were not described in any virus family so far. Phylogenetic analysis with all relevant representatives of the suggests that EMCLV should be nominated as a new species within the genus .
Topics: Animals; Female; Genome, Viral; High-Throughput Nucleotide Sequencing; Horse Diseases; Horses; Intercellular Signaling Peptides and Proteins; Membrane Proteins; Molluscipoxvirus; Molluscum contagiosum virus; Open Reading Frames; Phylogeny; Poxviridae Infections; Skin; Skin Diseases, Viral; Transcription, Genetic; Viral Proteins; Virus Replication; Whole Genome Sequencing
PubMed: 31922947
DOI: 10.1099/jgv.0.001357 -
Molecular Therapy. Methods & Clinical... Dec 2021Poxviruses have been used extensively as vaccine vectors for human and veterinary medicine and have recently entered the clinical realm as immunotherapies for cancer. We...
Poxviruses have been used extensively as vaccine vectors for human and veterinary medicine and have recently entered the clinical realm as immunotherapies for cancer. We present a comprehensive method for producing high-quality lots of the poxvirus (OrfV) for use in preclinical models of vaccinology and cancer therapy. OrfV is produced using a permissive sheep skin-derived cell line and is released from infected cells by repeated freeze-thaw combined with sonication. We present two methods for isolation and purification of bulk virus. Isolated virus is concentrated to high titer using polyethylene glycol to produce the final -grade product. We also describe methods for quantifying OrfV infectious virions and determining genomic copy number to evaluate virus stocks. The methods herein will provide researchers with the ability to produce high-quality, high-titer OrfV for use in preclinical studies, and support the translation of OrfV-derived technologies into the clinic.
PubMed: 34786436
DOI: 10.1016/j.omtm.2021.08.004 -
Journal of Veterinary Research Mar 2022Seal parapoxvirus (SPPV) infection has been reported among pinnipeds in aquaria in Japan; however, its seroprevalence is unknown. Therefore, an enzyme-linked...
INTRODUCTION
Seal parapoxvirus (SPPV) infection has been reported among pinnipeds in aquaria in Japan; however, its seroprevalence is unknown. Therefore, an enzyme-linked immunosorbent assay (ELISA) was developed for serological diagnosis of SPPV infection.
MATERIAL AND METHODS
The gene encoding the major envelope protein of SPPV was cloned into the eukaryotic expression vector pAcGFP1-N1, which encodes the green fluorescence protein (GFP), thereby producing a fusion protein (Env-GFP). Parental and cloned vector DNA was independently transfected into cultured seal cells for the expression of GFP and Env-GFP. The wells of an ELISA plate were coated with either GFP- or Env-GFP-transfected cell lysates. The light absorbance of each serum sample was adjusted by subtracting the absorbance of GFP-coated wells from that of Env-GFP-coated wells. Sera from two spotted seals (), six beluga whales (), three Pacific white-sided dolphins (), and ten bottlenose dolphins () from an aquarium in Japan were examined using the ELISA.
RESULTS
Positive reactions were not observed, except in one preserved sample collected ten years ago from a naturally SPPV-infected spotted seal.
CONCLUSION
The established ELISA could be useful in screening marine mammal sera for anti-SPPV antibodies.
PubMed: 35582482
DOI: 10.2478/jvetres-2022-0005