Canarypox Research Articles

A single dose of an ALVAC vector-based RABV virus-like particle candidate vaccine induces a potent immune response in mice, cats and dogs

Rabies, caused by the Rabies virus (RABV), is a highly fatal zoonotic disease. Existing rabies vaccines have demonstrated good immune efficacy, but the complexity of immunization procedures and high cost has impeded the elimination of RABV, particularly in the post-COVID-19 era. There is a pressing need for safer and more effective rabies vaccines that streamline vaccination protocols and reduce expense. To meet this need, we have developed a potential rabies vaccine candidate called ALVAC-RABV-VLP, utilizing CRISPR/Cas9 gene editing technology. This vaccine employs a canarypox virus vector (ALVAC) to generate RABV virus-like particles (VLPs). In mice, a single dose of ALVAC-RABV-VLP effectively activated dendritic cells (DCs), follicular helper T cells (Tfh), and the germinal center (GC)/plasma cell axis, resulting in durable and effective humoral immune responses. The survival rate of mice challenged with lethal RABV was 100%. Similarly, in dogs and cats, a single immunization with ALVAC-RABV-VLP elicited a stronger and longer-lasting antibody response. ALVAC-RABV-VLP induced superior cellular and humoral immunity in both mice and beagles compared to the commercial inactivated rabies vaccine. In conclusion, ALVAC-RABV-VLP induced robust protective immune responses in mice, dogs and cats, offering a novel, cost-effective, efficient, and promising approach for herd prevention of rabies.

Systemic avian poxvirus infections associated with the B1 subclade of canarypox virus.

Avian poxvirus infections typically manifest as 2 forms: cutaneous ("dry") pox, characterized by proliferative nodules on the skin, and diphtheritic ("wet") pox, characterized by plaques of caseous exudate in the oropharynx and upper respiratory and gastrointestinal tracts. Systemic spread of virus to visceral organs beyond the skin and mucous membranes is rarely reported. Out of 151 cases diagnosed with avian poxvirus over a 20-year period at a zoological institution, 22 were characterized as having systemic involvement based on histopathology and molecular findings. Gross lesions in systemic cases included soft white nodules scattered throughout the liver, spleen, and kidneys. Two histopathologic patterns emerged: (1) widespread histiocytic inflammation in visceral organs with intrahistiocytic viral inclusions and (2) severe, localized dry or wet pox lesions with poxvirus-like inclusions within dermal and subepithelial histiocytes. In situ hybridization targeting the core P4b protein gene confirmed the presence of poxvirus DNA within histiocytes in both patterns. Polymerase chain reaction was performed targeting the reticuloendothelial virus long terminal repeat (REV LTR) flanking region and the core P4b protein gene. Sequences of the REV LTR flanking region from all systemic pox cases were identical to a previously described condorpox virus isolated from an Andean condor with systemic pox. Sequences of the core P4b protein gene from all systemic pox cases grouped into cluster 2 of the B1 subclade of canarypox viruses. Systemic involvement of avian poxvirus likely occurs as a result of infection with certain strain variations in combination with various possible host and environmental factors.

Poxvirus infection in house finches (Haemorhous mexicanus): Genome sequence analysis and patterns of infection in wild birds.

Poxviruses (family: Poxviridae) infect many avian species, causing several disease outcomes, the most common of which are proliferative lesions on the legs, feet, and/or head. Few avian studies of poxvirus to date have combined molecular and ecological analyses to obtain a more comprehensive understanding of the identity and distribution of the disease in a population. Here, we describe patterns of poxvirus infection in an urban population of house finches (Haemorhous mexicanus) in Arizona (USA) and use high-throughput sequencing to determine the genome sequence of the virus. We found that poxvirus prevalence, based on visual identification of pox lesions, was 7.2% (17 infected birds out of a total of 235 sampled) in our population during summer 2021. Disease severity was low; 14 of the 17 infected birds had a single small lesion on the skin overlaying the eye, leg, and ear canal. All but two lesions were found on the feet; one bird had a lesion on the eye and the other in the ear opening. We also investigated possible temporal (i.e., date of capture) and biological correlates (e.g., age, sex, body condition, degree of infection with coccidian endoparasites) of poxvirus infection in urban-caught house finches during this time but found that none of these significantly correlated with poxvirus presence/absence. Two complete poxvirus genomes were determined from two infected birds. These genomes are ∼354,000bp and share 99.7% similarity with each other, and 82% with a canarypox virus genome, the most closely related avipoxvirus. This novel finchpox virus is the first to be reported in house finches and has a similar genome organization to other avipoxviruses.

Genomic Characterisation of a Novel Avipoxvirus Isolated from an Endangered Northern Royal Albatross (Diomedea sanfordi).

Marine bird populations have been declining globally with the factors driving this decline not fully understood. Viral diseases, including those caused by poxviruses, are a concern for endangered seabird species. In this study we have characterised a novel avipoxvirus, tentatively designated albatrosspox virus (ALPV), isolated from a skin lesion of an endangered New Zealand northern royal albatross (Diomedea sanfordi). The ALPV genome was 351.9 kbp in length and contained 336 predicted genes, seven of which were determined to be unique. The highest number of genes (313) in the ALPV genome were homologs of those in shearwaterpox virus 2 (SWPV2), while a further 10 were homologs to canarypox virus (CNPV) and an additional six to shearwaterpox virus 1 (SWPV1). Phylogenetic analyses positioned the ALPV genome within a distinct subclade comprising recently isolated avipoxvirus genome sequences from shearwater, penguin and passerine bird species. This is the first reported genome sequence of ALPV from a northern royal albatross and will help to track the evolution of avipoxvirus infections in this endangered species.

Another Promising HIV Vaccine Fails

The modest preventive efficacy seen in the RV144 vaccine trial in Thailand prompted an investigation of the recombinant canarypox vector (ALVAC-HIV)

Treatment of equine sarcoids using recombinant poxviruses expressing feline interleukin‐2

Interleukin (IL)-2 stimulates antitumour immunity and is successfully used for the treatment of different neoplasias. Canarypox virus locally expressing feline IL-2 is safe and can be used to treat equine sarcoids. Twenty horses of different breeds with a median age of eight years (interquartile range 6.0-13.3 years) and a total number of 59 sarcoids were included in the study. In this prospective clinical trial, sarcoids were injected twice seven days apart, with a recombinant canarypox virus expressing feline IL-2. Complete blood counts (CBC) and fibrinogen levels were measured before treatment and on days 1, 2, 7 and 8. Complete regression was achieved in eight horses (40%) and partial regression in two horses (10%). No change in sarcoid size was observed in two horses (10%) and the disease progressed in five horses (25%). Sarcoids of three horses (15%) showed initial response followed by tumour growth. There were no significant changes in CBC and fibrinogen levels after either injection. One horse developed a mild fever the day after each injection, which subsided without treatment the following day. Treatment of equine sarcoids with recombinant canarypox virus expressing feline IL-2 seems to be a safe therapy option. Although the expression of IL-2 after vector injection and its biological activity in horses were not proven in this study, the treatment resulted in regression and partial regression in 50% of the cases. Further studies are necessary to verify these findings and to establish a treatment protocol.

A highly efficient recombinant canarypox virus-based vaccine against canine distemper virus constructed using the CRISPR/Cas9 gene editing method

Canine distemper virus (CDV) is the causative agent of canine distemper (CD), which is one of the most important infectious diseases affecting wild and domestic carnivores. Vaccination represents an effective approach to prevent CDV infection among domestic carnivores. Canarypox-vectored recombinant CD vaccines (such as Recombitek CDV, PureVax Ferret Distemper, and Merial) with the CDV hemagglutinin (H) and fusion (F) genes can induce a potent immune response in dogs and ferrets. However, the vaccine’s effectiveness varies with the species. In the current study, we developed a highly efficient recombinant canarypox virus termed as “ALVAC-CDV-M-F-H/C5−” that contained CDV virus-like particles (VLPs) by using the CRISPR/Cas9 gene editing method, which enabled concurrent expression of the matrix (M), H, and F genes. The recombinant strain provided faster seroconversion than the parent strain among minks as well as provided higher rates of antibody positivity than the parent strain among foxes and minks even before the administration of a second booster vaccination. We demonstrated, for the first time, that the CRISPR/Cas9 system can be applied for the rapid and efficient modification of the ALVAC-CDV-F-H genome and also that a high-dose new recombinant strain that produces CDV VLPs may present good outcomes in the prevention of CD among foxes and minks.

Molecular Detection of Reticuloendotheliosis Virus 5' Long Terminal Repeat Integration in the Genome of Avipoxvirus Field Strains from Different Avian Species in Egypt.

Avipoxviruses (APVs) are among the most complex viruses that infect a wide range of birds’ species. The infection by APVs is often associated with breathing and swallowing difficulties, reduced growth, decreased egg production, and high mortalities in domestic poultry. In the present study, 200 cutaneous nodular samples were collected from different avian species (chicken, pigeon, turkey, and canary) suspected to be infected with APVs from Dakahlia Governorate, Egypt. Pooled samples (n = 40) were prepared and inoculated in embryonated chicken eggs (ECEs). APVs were then identified by polymerase chain reaction (PCR) and sequence analysis of the APV P4b gene. Furthermore, the forty strains of APVs were screened for the presence of reticuloendotheliosis virus (REV)-5′LTR in their genomes. Interestingly, the phylogenic tree of the APV P4b gene was separated into 2 clades: clade 1, in which our fowlpox virus (FWPV), turkeypox virus (TKPV), and canarypox virus (CNPV) isolates were grouped, along with reference FWPVs and TKPVs retrieved from GenBank, whereas, in clade2, the pigeonpox virus (PGPV) isolate was grouped with PGPVs retrieved from GenBank. Likewise, REV-5′LTR was amplified from 30 strains isolated from chicken, turkey, and canary, while PGPV strains were free from REV-5′LTR integration. To the best of our knowledge, this study involved the detection and characterization of REV-5′LTR insertions in the APVs field isolates in Egypt for the first time. Given the above information, further future research seems recommended to understand the impact of the resulting REV-5′LTR insertions on the pathogenesis, virulence, and inadequate vaccine protection against APVs.

WHOLE GENOME SEQUENCING OF AN AVIPOXVIRUS ASSOCIATED WITH INFECTIONS IN A GROUP OF AVIARY-HOUSED SNOW BUNTINGS (PLECTROPHENAX NIVALIS).

Avipoxvirus infections have been reported in both free-ranging and domestic birds worldwide. Fowlpox and canarypox viruses belong to the genus Avipoxvirus among the virus family Poxviridae. They cause cutaneous lesions with proliferative growths on the unfeathered parts of the skin and/or diphtheritic lesions generally associated with necrosis in the upper respiratory and digestive tracts. In this study, a poxvirus has been identified in wild-caught snow buntings (Plectrophenax nivalis) housed in an outdoor aviary in the region of Rimouski, Quebec. During the falls and winters of 2015 and 2016, eight snow buntings affected by this infection were examined. Macroscopic and microscopic lesions observed were characteristic of an avipoxvirus infection. Electron microscopy imaging of an ultrathin section of the histopathological lesions of two birds confirmed the presence of the poxvirus. Afterward, the presence of the poxvirus was confirmed in three birds by a specific polymerase chain reaction assay that amplified a segment of the gene encoding the fowlpox virus 4b core protein. A 576-nucleotide amplicon was obtained from one of them and sequenced. The analyses revealed a 99% homology to other previously described avipoxviruses. Using high-throughput sequencing, almost the entire viral genome of this avipoxvirus was revealed and found to possess a 359,853-nucleotide sequence in length. Bioinformatic analyses revealed that the virus was genetically related to canarypox virus. To our knowledge, this is the first confirmed case and full description of a poxviral infection in this species. This episode suggests a high susceptibility of this northern species of passerine to avipoxviruses circulating in southeastern Canada during the summer months. Even if the source of the viral infections remains undetermined, transmission by local biological vectors is suspected. Management of poxviral infections in snow buntings housed outdoors in southeastern Canada could rely on the control of biting insects.

Simplified steps to heterologous prime-boost HIV vaccine development?

The RV 144 HIV vaccine efficacy study showed a reduction in HIV-1 infection risk in Thai volunteers who received two priming vaccinations of vCP1521 ALVAC (attenuated recombinant canarypox virus expressing HIV group-specific antigen, polymerase, and envelope genes) followed by two additional ALVAC vaccinations and coadministration of purified bivalent gp120 proteins (AIDSVAX B/E). In this issue of the JCI, Rouphael et al. build on these results by substituting a DNA plasmid cocktail expressing HIV-1 subtype C group-specific antigen, polymerase, and envelope antigen genes (DNA-HIV-PT123) for ALVAC in a phase 1b safety and immunogenicity study. The results indicate that the vaccine regimen is safe, elicits promising cross-subtype humoral and cellular responses, and opens up potentially simplified approaches to HIV-1 vaccine development.

Molecular Phylogeny of an Avipoxvirus Isolated from Red-Flanked Blue Robin in China.

We first report avipoxvirus (APV) infection and an isolate named APV/03/2016 from a red-flanked blue robin (Tarsiger cyanurus) captured at Songhua Lake Scenic Area in Jilin City (Jilin Province, China) on March 24, 2016. The partial sequence of the 4b core protein gene and DNA polymerase gene of APV/03/2016 suggests that the virus belongs to the subclade B1 cluster of clade B (canarypox virus). The BLAST results showed the highest similarity of the two genes with the Pacific shearwater-isolated strain SWPV-2 (KX857215), canarypox virus strain D98-11133 (GQ487567), canarypox virus strain ATCC VR-111 (AY318871), avipoxvirus Mississippi isolate P89 (KC018048), and avipoxvirus Wisconsin isolate P92 (KC018051). The results indicate that APV/03/2016 is a canarypox-like virus. These findings demonstrate the continuous emergence of new APV hosts such as red-flanked blue robins and suggest that monitoring of APV circulation and evolution should be strengthened for T. cyanurus conservation.

Characterization of Iranian canarypox and pigeonpox virus strains.

Avipoxviruses (APVs) are large DNA viruses that are detected widely in many species of birds. Little information is available regarding genetic variations in these host-specific viruses. In the present study, nine canarypox virus and five pigeonpox virus isolates were collected from northeastern Iran and isolated via the chorioallantoic membrane of chicken embryos. Further investigations were conducted using analysis of virus growth in chicken embryo fibroblasts, histopathology, electron microscopy, and molecular techniques such as polymerase chain reaction (PCR) combined with sequencing and phylogenetic analysis to investigate variations in the highly conserved P4b gene of poxviruses. Virus replication and pock lesions were evident, and microscopic examination revealed eosinophilic intracytoplasmic inclusion bodies and biconcave enveloped virus particles with randomly arranged surface filaments, which are characteristic features of poxviruses. PCR results confirmed the presence of an APV-specific 578-bp fragment in all of the samples. Sequence analysis and phylogenetic analysis of 578-bp P4b fragments of eight isolates confirmed that our canary and pigeon isolates clustered with previously reported isolates. The similarity between the nucleotide sequences of most of our isolates and those isolated previously in other countries could be due to the high degree of conservation of these fragments. However, the FZRC6V isolate from a canary in this study did not have a canarypox virus origin according to the sequence analysis, and might have originated from cross-infection with different strains of avipoxviruses.

A flow cytometric granularity assay for the quantification of infectious virus

A flow cytometry-based assay was developed to assess the infective titer of two recombinant viruses: a recombinant herpes simplex type 2 (rHSV-2) and a recombinant canary pox (rALVAC.gfp). This method uses granularity of infected Vero and QT-35 cells, respectively, and correlates this to the infectious titer of virus samples. The percent of the cell populations with a high level of granularity could accurately be correlated to viral titers obtained through a traditional plaque assay, with R2 values greater than 0.8 using a semi-logarithmic scale. This approach offers a rapid, high-throughput method for infectious virus titration with similar accuracy to a traditional plaque assay.

Immunological response in horses following West Nile virus vaccination with inactivated or recombinant vaccine.

To evaluate the immunological response following vaccination, 40 WNV serologically negative horses were selected and divided in two groups of 20 animals. One group was vaccinated (booster after 28 days) with a whole inactivated viral strain and the second group with a live recombinant canarypox virus expressing the genes coding for the WNV preM/E viral proteins. IgM, IgG and neutralizing antibodies were monitored by class specific ELISAs and serum neutralization assay for 360 days. In both groups, IgM antibodies were first detected 7 days post vaccination (dpv). However, in the group vaccinated with inactivated vaccine, IgM antibodies were detected until day 42 pv, whereas in the group vaccinated with the recombinant vaccine, they were detected up to day 52 pv. A similar (P > 0.05) proportion of horses showed IgM antibodies after vaccination with either recombinant [30%; 95% confidence interval (CI): 14.59%-52.18%] or inactivated (32%; 95% CI: 15.39-54.28%) vaccine. Both vaccines induced in vaccinated horses a detectable IgG antibody response starting from day 7 pv and lasting till the end of the trial. Analogously, both products elicited WNV specific neutralizing antibodies. The response induced by the live canarypox virus-vectored vaccine was higher (mean titres 1:298 vs 1:18.9) and lasted longer than did that induced by the killed-virus vaccines. In fact, one year after the vaccination, neutralizing antibodies were still detectable in the horses which received the canarypox virus-based vaccine but not in the group vaccinated with the killed product. The use of vaccines is a valuable tool to prevent WNV disease in horses and the availability of different products facilitates the control of the disease in endemic areas.

Chicken Embryonic-Stem Cells Are Permissive to Poxvirus Recombinant Vaccine Vectors.

The discovery of mammalian pluripotent embryonic stem cells (ESC) has revolutionised cell research and regenerative medicine. More recently discovered chicken ESC (cESC), though less intensively studied, are increasingly popular as vaccine substrates due to a dearth of avian cell lines. Information on the comparative performance of cESC with common vaccine viruses is limited. Using RNA-sequencing, we compared cESC transcriptional programmes elicited by stimulation with chicken type I interferon or infection with vaccine viruses routinely propagated in primary chicken embryo fibroblasts (CEF). We used poxviruses (fowlpox virus (FWPV) FP9, canarypox virus (CNPV), and modified vaccinia virus Ankara (MVA)) and a birnavirus (infectious bursal disease virus (IBDV) PBG98). Interferon-stimulated genes (ISGs) were induced in cESC to levels comparable to those in CEF and immortalised chicken fibroblast DF-1 cells. cESC are permissive (with distinct host transcriptional responses) to MVA, FP9, and CNPV but, surprisingly, not to PBG98. MVA, CNPV, and FP9 suppressed innate immune responses, while PBG98 induced a subset of ISGs. Dysregulation of signalling pathways (i.e., NFκB, TRAF) was observed, which might affect immune responses and viral replication. In conclusion, we show that cESC are an attractive alternative substrate to study and propagate poxvirus recombinant vaccine vectors.

New generation vaccines against bluetongue virus

Over the last three decades, a variety of approaches have been investigated to develop new types of bluetongue virus (BTB) vaccines, ranging from baculovirus-expressed subunit vaccines to live vector vaccines. DNA vaccines against BTV consist of DNA plasmid expressing different BTV proteins after inoculation of the animals. The recombinant viral vector vaccines against BTV are based on recombinant viruses that express desired BTV antigens in the host upon inoculation. Viruses such as vaccinia, modified vaccinia Ancara (MVA), capripox, canarypox, herpes, myxoma and fowlpox viruses have been used as vectors of BTV genes. The reverse genetics (RG) systems for BTV are useful tools for BTV vaccine development. Disabled infectious single-cycle (DISC) vaccines make it possible to restore virus replication and can be used for differentiating infected from vaccinated animals (DIVA). These vaccines are based on the production of a modified virus with a deletion in one or more genes that are essential for virus replication. Another approach for BTV vaccine development using RG is the disabled infectious single-animal (DISA) vaccine, generated by deletion of NS3/NS3a expression. DISC and DISA vaccines can mimic the natural tropism of the virus and can express BTV proteins at the site of infection. Important advantages of these new generation vaccines over the conventional BTV vaccines are their high efficacy as well as the possibility of applying them for DIVA. At present, there are a number of novel laboratory-scale BTV vaccines that could meet vaccine profiles required for different field situations. However, further development and licensing of these vaccine candidates for many BTV serotypes is needed in order to prepare for future BT outbreaks. To date, all novel BTV vaccines described in this paper are still under laboratory testing. They are not available commercially, and the time of their application in the field is still indefinite. .

Characterization of a canarypox virus from an outbreak among canaries (Serinus canaria domesticus) in Lebanon

ABSTRACTA canarypox outbreak resulted in the death of about 50% of canaries (Serinus canaria domesticus) in several breeding farms in Lebanon. Infected birds showed thickened eye lid and skin scab-like lesions at the beak, foot and caudal regions and died 5–6 days post disease symptoms onset. Out of seven sick birds that were autopsied for gross pathology evaluation, one bird demonstrated turbid airsacs with absence of any lesions in the trachea, oesophagus, liver and lung. Histopathological examination showed hyperplasia, heterophils infiltration and Bollinger body formation in the skin of five out of the seven autopsied canaries. Hyperplasia and infiltration of heterophils were also observed in the airsacs of one bird. PCR analysis of specimens taken from the skin and feet, targeting the fpv167 gene of the canarypox virus, was positive for all of the analyzed canaries. PCR analysis also revealed that two birds had concurrent infection with Mycoplasma gallisepticum. Sequencing and alignment of the amplified fpv167 gene of the canarypox showed 100% similarity with the Iranian canary pox isolate IR/H913/14. Smuggling of pet birds through the borders should be strictly controlled and biosecurity measures must be adequately applied to control the circulation of the two identified pathogens.

Priming and Activation of Inflammasome by Canarypox Virus Vector ALVAC via the cGAS/IFI16-STING-Type I IFN Pathway and AIM2 Sensor.

Viral vectors derived from different virus families, including poxvirus (canarypox virus vector ALVAC) and adenovirus (human Ad5 vector), have been widely used in vaccine development for a range of human diseases including HIV/AIDS. Less is known about the mechanisms underlying the host innate response to these vectors. Increasing evidence from clinical vaccine trials testing different viral vectors has suggested the importance of understanding basic elements of host-viral vector interactions. In this study, we investigated the innate interactions of APCs with two commonly used HIV vaccine vectors, ALVAC and Ad5, and identified AIM2 as an innate sensor for ALVAC, triggering strong inflammasome activation in both human and mouse APCs. Microarray and comprehensive gene-knockout analyses (CRISPR/Cas9) identified that ALVAC stimulated the cGAS/IFI16-STING-type I IFN pathway to prime AIM2, which was functionally required for ALVAC-induced inflammasome activation. We also provided evidence that, in contrast to ALVAC, the Ad5 vector itself was unable to induce inflammasome activation, which was related to its inability to stimulate the STING-type I IFN pathway and to provide inflammasome-priming signals. In preconditioned APCs, the Ad5 vector could stimulate inflammasome activation through an AIM2-independent mechanism. Therefore, our study identifies the AIM2 inflammasome and cGAS/IFI16-STING-type I IFN pathway as a novel mechanism for host innate immunity to the ALVAC vaccine vector.

Structural and Functional Insight into Canarypox Virus CNP058 Mediated Regulation of Apoptosis

Programmed cell death or apoptosis is an important component of host defense systems against viral infection. The B-cell lymphoma 2 (Bcl-2) proteins family is the main arbiter of mitochondrially mediated apoptosis, and viruses have evolved sequence and structural mimics of Bcl-2 to subvert premature host cell apoptosis in response to viral infection. The sequencing of the canarypox virus genome identified a putative pro-survival Bcl-2 protein, CNP058. However, a role in apoptosis inhibition for CNP058 has not been identified to date. Here, we report that CNP058 is able to bind several host cell pro-death Bcl-2 proteins, including Bak and Bax, as well as several BH3 only-proteins including Bim, Bid, Bmf, Noxa, Puma, and Hrk with high to moderate affinities. We then defined the structural basis for CNP058 binding to pro-death Bcl-2 proteins by determining the crystal structure of CNP058 bound to Bim BH3. CNP058 adopts the conserved Bcl-2 like fold observed in cellular pro-survival Bcl-2 proteins, and utilizes the canonical ligand binding groove to bind Bim BH3. We then demonstrate that CNP058 is a potent inhibitor of ultraviolet (UV) induced apoptosis in a cell culture model. Our findings suggest that CNP058 is a potent inhibitor of apoptosis that is able to bind to BH3 domain peptides from a broad range of pro-death Bcl-2 proteins, and may play a key role in countering premature host apoptosis.

Genomic characterization of two novel pathogenic avipoxviruses isolated from pacific shearwaters (Ardenna spp.)

BackgroundOver the past 20 years, many marine seabird populations have been gradually declining and the factors driving this ongoing deterioration are not always well understood. Avipoxvirus infections have been found in a wide range of bird species worldwide, however, very little is known about the disease ecology of avian poxviruses in seabirds. Here we present two novel avipoxviruses from pacific shearwaters (Ardenna spp), one from a Flesh-footed Shearwater (A. carneipes) (SWPV-1) and the other from a Wedge-tailed Shearwater (A. pacificus) (SWPV-2).ResultsEpidermal pox lesions, liver, and blood samples were examined from A. carneipes and A. pacificus of breeding colonies in eastern Australia. After histopathological confirmation of the disease, PCR screening was conducted for avipoxvirus, circovirus, reticuloendotheliosis virus, and fungal agents. Two samples that were PCR positive for poxvirus were further assessed by next generation sequencing, which yielded complete Shearwaterpox virus (SWPV) genomes from A. pacificus and A. carneipes, both showing the highest degree of similarity with Canarypox virus (98% and 67%, respectively). The novel SWPV-1 complete genome from A. carneipes is missing 43 genes compared to CNPV and contains 4 predicted genes which are not found in any other poxvirus, whilst, SWPV-2 complete genome was deemed to be missing 18 genes compared to CNPV and a further 15 genes significantly fragmented as to probably cause them to be non-functional.ConclusionThese are the first avipoxvirus complete genome sequences that infect marine seabirds. In the comparison of SWPV-1 and −2 to existing avipoxvirus sequences, our results indicate that the SWPV complete genome from A. carneipes (SWPV-1) described here is not closely related to any other avipoxvirus genome isolated from avian or other natural host species, and that it likely should be considered a separate species.