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In September 2023, five semi-domesticated reindeer calves ( Rangifer tarandus tarandus ) from Norrbotten County, Sweden, displayed clinical signs resembling contagious ecthyma. Samples were collected from the lesion sites, including the skin of the muzzle, eyelids, and oral mucosa. The samples were analysed via real-time PCR (qPCR) for parapoxviruses (PPV) and cervidpoxviruses (CvPV). Both viruses were detected in samples from several calves. Full genome sequencing of the PPV strain 23-MIK191411 revealed close similarity to the parapoxvirus red deer (RDPV) strain HL953 from a red deer ( Cervus elaphus ) in Germany but with a novel 2,187 bp insert in the right terminal third of the genome. This insert carried two open reading frames (ORFs) that, while divergent, presented sequence homology with several other PPVs. Analysis of amino acid differences relative to the sequence of the RDPV HL953 strain revealed that proteins implicated in host interactions and virulence presented the greatest differences. Thus, comparative sequence analysis indicates that an RDPV has recombined with an ancestor of 23-MIK191411 and adapted to the reindeer host. This study represents the first detection of RDPV, with a unique insert likely originating from an unknown PPV in reindeer, indicating its emergence beyond the red deer host range. Clinical signs are consistent with those caused by other poxviruses, including CvPV also detected in the investigated animal group, making it difficult to diagnose the causative agent solely via clinical observation. In the sequenced sample, RDPV was confirmed as the predominant variant. These findings underscore the importance of molecular diagnostics for accurate pathogen identification and highlight the need for continued health surveillance of reindeer. Further investigations are needed to determine the clinical impact of RDPV in reindeer.

Background Lumpy Skin Disease (LSD) is endemic in sub-Saharan countries and is currently a global threat to the cattle industry. Information on the circulating Capripoxvirus lumpyskinpox, formerly known as Lumpy Skin Disease Virus (LSDV), and other poxviruses infecting cattle is very scant in Tanzania. The current study aimed to confirm and characterize LSDV and other poxviruses infecting cattle, from LSD suspected outbreaks in Tanzania. Methods A total of 24 samples were collected from four LSD suspected outbreaks reported in Tanzania between February and May 2023. Samples were screened for LSDV genome by real-time PCR and then subjected to a high-resolution multiplex melting (HRM) assay where 10 samples were positive for Capripoxvirus (CaPV) and one sample was Parapoxvirus (PPV) positive. Four LSDV genes; RPO30, GPCR, EEV glycoprotein and B22R and the partial B2L gene of PPVs were analyzed. Results All targeted LSDV genes from the Tanzanian isolates showed 100% similarity and isolates clustered with commonly circulating LSDV field isolates. Furthermore, the single nucleotide polymorphism (SNP) at position 240 (A-> G) of the EEV gene differentiates the Tanzanian LSDVs from the group of ancient Kenyan LSDV isolates while the B22R sequences of the Tanzanian LSDV isolates differed from the LSDV Neethling and LSDV KSGP-0240 derived vaccines. Sequence analysis of the partial B2L gene of the Tanzanian parapoxvirus bovinestomatitis, formerly known as Bovine papular stomatitis virus (BPSV) showed a different BPSV strain circulating compared to publicly available sequences. Conclusion These findings confirm the presence of LSDV in Tanzania, which suggesting the need for establishing an effective control program and continuous monitoring. The presence of a typical profile for Tanzania BPSV is an indication that, although never reported before, BPSV is established in the country therefore this virus should be included in the differential diagnosis of LSDV.

We report a sequencing protocol and 121-kb poxvirus sequence from a clinical sample from a horse in Finland with dermatitis. Based on phylogenetic analyses, the virus is a novel parapoxvirus associated with a recent epidemic; previous data suggest zoonotic potential. Increased awareness of this virus and specific diagnostic protocols are needed.

Two cases of coinfection with bovine papular stomatitis virus (BPSV) and pseudocowpox virus (PCPV) in dairy calves in Tochigi Prefecture, Japan, are reported. Sequences of BPSV and PCPV were simultaneously detected in the same polymerase chain reaction (PCR) amplicons, which were obtained from the DNA of two dairy calves using a pan-parapoxvirus primer set. PCR amplification using BPSV- and PCPV-specific primer sets were able to distinguish between the two viruses in coinfected clinical samples. Based on these data, further studies on the occurrence BPSV/PCPV coinfections in cattle in Japan are warranted.

The advantage of oncolytic viruses (OV) in cancer therapy is their dual effect of directly killing tumours while prompting anti-tumour immune response. Oncolytic parapoxvirus ovis (ORFV) and other OVs are thought to induce apoptosis, but apoptosis, being the immunogenically inert compared to other types of cell death, does not explain the highly inflamed microenvironment in OV-challenged tumors. Here we show that ORFV and its recombinant therapeutic derivatives are able to trigger tumor cell pyroptosis via Gasdermin E (GSDME). This effect is especially prominent in GSDME-low tumor cells, in which ORFV-challenge pre-stabilizes GSDME by decreasing its ubiquitination and subsequently initiates pyroptosis. Consistently, GSDME depletion reduces the proportion of intratumoral cytotoxic T lymphocytes, pyroptotic cell death and the success of tumor ORFV virotherapy. In vivo, the OV preferentially accumulates in the tumour upon systemic delivery and elicits pyroptotic tumor killing. Consequentially, ORFV sensitizes immunologically ‘cold’ tumors to checkpoint blockade. This study thus highlights the critical role of GSDME-mediated pyroptosis in oncolytic ORFV-based antitumor immunity and identifies combinatorial cancer therapy strategies. Oncolytic viruses are able to target tumours and thought to induce apoptosis while remodelling the tumour immune microenvironment. Here authors show in an oncolytic parapoxvirus ovis model that pyroptosis, a highly immunogenic Gasdermin-E-dependent cell death mechanism, is the dominant cell death pathway during virotherapy.

Outbreaks of emerging diseases, like Mpox in 2022, pose unprecedented challenges to global healthcare systems. Although Mpox cases globally decreased since the end of 2022, numbers are still significant in the African Region, European Region, Region of the Americas, and Western Pacific Region. Rapid and efficient detection of infected individuals by precise screening assays is crucial for successful containment. In these assays, analytical and clinical performance must be assessed to ensure high quality. However, clinical studies evaluating Mpox virus (MPXV) detection kits using patient-derived samples are scarce. This study evaluated the analytical and clinical performance of a new diagnostic MPXV real-time PCR detection kit (Sansure Monkeypox Virus Nucleic Acid Diagnostic Kit) using patient-derived samples collected in Germany during the MPXV clade IIb outbreak in 2022. Our experimental approach determined the Limit of Detection (LoD) to less than 200 cp/mL using whole blood samples and samples derived from vesicles or pustules. Furthermore, we tested potentially inhibiting substances and pathogens with homologous nucleic acid sequences or similar clinical presentation and detected no cross-reactivity or interference. Following this, the assay was compared to a CE-marked test in a clinical performance study and achieved a diagnostic sensitivity of 100.00% and diagnostic specificity of 96.97%. In summary, the investigated real-time PCR assay demonstrates high analytical performance and concurs with the competitor device with high specificity and sensitivity.

Bovine papular stomatitis virus (BPSV) is a parapoxvirus that infects cattle, causing skin lesions on the udder and mouth. There have been few studies on the prevalence and molecular characteristics of BPSV in Iraq. Here, we describe the prevalence, phylogenetic analysis, and clinico-epidemiological features of BPSV in cattle in Al-Qadisiyah, Iraq. A total of 264 animals were examined for teat and oral lesions, and BPSV was detected by PCR in 79.9% (211/264) of cattle and calves with skin lesions. The lesions included ulcers, papules, and scabby proliferative areas. The BPSV strains from Iraq clustered phylogenetically with BPSV strains detected in the USA. Further studies are needed to explore the evolution and epidemiology of this virus in the region.

Two deer hunters presented with a violaceous nodule on a finger several weeks after field-dressing white-tailed deer. Electron microscopy revealed ovoid virions suggestive of parapoxvirus. DNA sequence analysis indicated that the causative agent was a unique parapoxvirus. In early November 2008, a 52-year-old wildlife biologist was deer hunting in eastern Virginia when he nicked his right index finger while dressing a white-tailed deer ( Odocoileus virginianus ). The deer appeared to be healthy at the time of death, with no lesions on its muzzle or head. The cut on the hunter's finger did not heal, and within 2 weeks, a tender raised area had begun to form at the wound site. The hunter reported having had no pain or itching at the site and noted no other lesions. He had no fever or other symptoms of systemic illness. By the second week of December 2008, the lesion at the wound site had enlarged to form a violaceous nodule (Figure 1A), and the patient sought medical attention.

Orf virus (ORFV) is a typical member of the genus Parapoxvirus. The parapoxvirus genome consists of highly variable terminal regions and relatively conserved central regions with a high G + C content. In our previous study, a novel ORFV strain, NA1/11, was isolated from northeastern China. To fully characterize this strain, we sequenced the entire genome of NA1/11 and conducted a comparative analysis using multiple parapoxviruses. The genomic sequence of NA1/11 was found to consist of 137,080 nucleotides with a G + C content of 63.6 %, but it did not contain the terminal hairpin sequence. Alignment of ORFs from NA1/11 with NZ2, IA82 and SA00 revealed several highly variable ORFs, while the most evident ones are ORFs 001, 103, 109–110, 116 and 132. An odd phenomenon in the region of ORFs 118–120 is that the non-coding fragments are almost as long as the coding fragments. By comparative analysis of inverted terminal repeats, we identified one repeat motif and a long conserved fragment. By comparing the ITRs of SA00 with those of three other ORFVs, more clues were obtained about the correlation between ITR sequence and host adaption. Comparison of the NA1/11 genome with the sequences of other strains of ORFV revealed highly variable regions, thus providing new insights into the genetic diversity of ORFV.

Foot and mouth disease (FMD) is an acute viral disease in animals. Inactive parapoxvirus ovis (IPPVO) strengthens humoral immunity. This study aimed to determine the effect of IPPVO application together with FMD vaccine on cattle immunity. It included 30 Holstein cattle randomly divided into two groups: one was administered only the FMD vaccine, and the other was administered the FMD vaccine and IPPVO simultaneously. Control blood was collected from all animals at 0 hours. Serum TNF-a, IL-1(3, and IL-6 levels were measured in blood samples collected at 4, 8, 12, 16, and 24 hours post-vaccination, while serum IgG and IgM levels were measured in blood samples collected at 4, 8, 12, 16, and 20 days post-vaccination using ELISAkits. While no changes in serum TNF-a, IL-1(3, and IL-6 levels were detected in the FMD group, IL-1(3 levels significantly increased (peaking at four hours) in the FMD + IPPVO group. In the FMD group, while IgG levels increased significantly (peaking at 16 days), IgM levels did not change. In the FMD + IPPVO group, IgG level on day 8 days was higher than the 16 days value. Also, the IgM level increased significantly on day 16. In conclusion, the application of FMD with IPPVO increases the primary immune response (IgM), but it does not effect the long-term immune response (IgM).