Human orf virus (family Poxviridae) infection following a lamb bite in Hungary.

Human orf disease (called ecthyma contagiosum or contagious/infectious pustular dermatitis in animals) was confirmed on the fingers of both hands of a 24-year-old female, after feeding diseased lambs with a nursing bottle in April 2023. In addition to skin symptoms, she had low-grade fever (37.6°C) and swollen lymph nodes in both axilla. The presence of orf virus (genus Parapoxvirus, family Poxviridae) was confirmed, and this strain, Baja/2023/HUN (OR372161-OR372163), was found to have > 98% nucleotide sequence identity to sheep-origin orf viruses in four tested genome regions (ORF011/B2L, ORF019, ORF020/VIR, and ORF056). This is the first report of a human case of infection with the neglected zoonotic orf virus in Hungary.

Detection and genetic characterization of orf virus from sheep and goats in Nigeria.

Orf is a contagious, viral epitheliotropic disease of small ruminants. We investigated the molecular epidemiology of orf virus (ORFV) in breeds of small ruminants to determine the evolutionary diversity in Nigeria. Out of 54 small ruminants screened, the number of animals that were positive for ORFV in the three locations were 25. The distribution of positive animals by location were FCT 45.0% (n = 9/20), Oyo State 42.9% (6/14), and Plateau State 50.0% (n = 10/20). ORFV sequences from this study clustered with viruses detected in Taiwan, Iran, USA, and France. Our findings highlight the risk of transmission across geographic boundaries in Nigeria and West Africa, and reinforces the need for increased surveillance to prevent and control spread. Comprehensive characterization of ORFV in small ruminants as well as in humans in Nigeria is required to better elucidate the epidemiological dynamics and the virus evolution.

The Global Evolutionary History of Orf Virus in Sheep and Goats Revealed by Whole Genomes Data.

Orf virus (ORFV) belongs to the genus Parapoxvirus (Poxviridae family). It is the causative agent of contagious ecthyma (CE) that is an economically detrimental disease affecting small ruminants globally. Contagious ecthyma outbreaks are usually reported in intensive breeding of sheep and goats but they have also been reported in wildlife species. Notably, ORFV can infect humans, leading to a zoonotic disease. This study aims to elucidate the global evolutionary history of ORFV genomes in sheep and goats, including the first genomes from Central America in the analyses. In comparison to the last study on ORFV whole genomes, the database now includes 11 more sheep and goat genomes, representing an increase of 42%. The analysis of such a broader database made it possible to obtain a fine molecular dating of the coalescent time for ORFV S and G genomes, further highlighting the genetic structuring between sheep and goat genomes and corroborating their emergence in the latter half of 20th century.

Genetic Analysis of Orf Virus (ORFV) Strains Isolated from Goats in China: Insights into Epidemiological Characteristics and Evolutionary Patterns.

Contagious ecthyma (CE) is an acute infectious zoonosis caused by orf virus (ORFV) that mainly infects sheep and goats and causes obvious lesions and low market value of livestock, resulting in huge economic losses for farmers. In this study, two strains of ORFV were isolated from Shaanxi Province and Yunnan Province in China, named FX and LX. The two ORFVs were located in the major clades of domestic strains respectively, and exhibited distinct sequence homology. We analyzed the genetic data of core genes (B2L, F1L, VIR, ORF109) and variable genes (GIF, ORF125 and vIL-10) of ORFV to investigate its epidemiological and evolutionary characteristics. The sequences from 2007 to 2018 constituted the majority of the viral population, predominantly concentrated in India and China. Most genes were clustered into SA00-like type and IA82-like type, and the hotspots in East and South Asia were identified in the ORFV transmission trajectories. For these genes, VIR had the highest substitution rate of 4.85 × 10-4, both VIR and vIL-10 suffered the positive selection pressure during ORFV evolution. Many motifs associated with viral survival were distributed among ORFVs. In addition, some possible viral epitopes have been predicted, which still require validation in vivo and in vitro. This work gives more insight into the prevalence and phylogenetic relationships of existing orf viruses and facilitate better vaccine design.

Host range, severity and trans boundary transmission of Orf virus (ORFV).

Contagious ecthyma in small ruminants is a zoonotic disease caused by Orf virus (ORFV) in the genus Parapoxvirus that can be deadly to its natural hosts. It causes significant losses worldwide, and commonly infects humans. However, the literature about its comparative severity in sheep and goat hosts is misleading; and while contagious ecthyma has been shown to occur in camels and transmit to humans, there is confusion as to whether ORFV is responsible. Camels are important from a 'One Health' perspective as they have been implicated as a reservoir host for the virus causing Middle East Respiratory Syndrome (MERS), which has a case fatality rate of 35% in humans. We compared ORFV gene sequences and mortality data from the West Bank in Palestine, where ORFV has not been reported previously, with data from the region. Surprisingly, we found that infections of camels that had been attributed to ORFV were more closely related to a different member of the genus Parapoxvirus. Two Middle East ORFVs isolated from humans were unrelated and sat alongside sheep and goat derived sequences on two distinct ORFV lineages of a maximum likelihood B2L gene tree. One of the viral lineages bifurcated to produce a monophyletic group of goat-derived ORFVs characterized uniquely by a glycine at amino acid reside 249. We found that serine is the ancestral allele shared between ORFV infections of sheep and also two closely related Parapoxviruses (PCPV and CCEV), indicating that the glycine allele represents a more recent shift in virus host range adaptation to goats. Furthermore, and contrary to some reports that ORFV is more severe in goats than in sheep, we observed median mortality of up to 24.5% in sheep, but none in goats. We also identified trans-boundary spread of ORFV between the West Bank and Israel.

Orf Nodule with Erythema Multiforme during a Monkeypox Outbreak, France, 2022.

A 26-year-old patient in France who worked as a butcher sought care initially for erythema multiforme. Clinical examination revealed a nodule with a crusty center, which upon investigation appeared to be an orf nodule. Diagnosis was confirmed by PCR. The patient was not isolated and had a favorable outcome after basic wound care.

Recent advances in diagnostic approaches for orf virus.

Orf virus (ORFV), the prototype species of the Parapoxvirus genus, is an important zoonotic virus, causing great economic losses in livestock production. At present, there are no effective drugs for orf treatment. Therefore, it is crucial to develop accurate and rapid diagnostic approaches for ORFV. Over decades, various diagnostic methods have been established, including conventional methods such as virus isolation and electron microscopy; serological methods such as virus neutralization test (VNT), immunohistochemistry (IHC) assay, immunofluorescence assay (IFA), and enzyme-linked immunosorbent assay (ELISA); and molecular methods such as polymerase chain reaction (PCR), real-time PCR, loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA), and recombinase-aided amplification (RAA) assay. This review provides an overview of currently available diagnostic approaches for ORFV and discusses their advantages and limitations and future perspectives, which would be significantly helpful for ORFV early diagnosis and surveillance to prevent outbreak of orf. KEY POINTS: • Orf virus emerged and reemerged in past years • Rapid and efficient diagnostic approaches are needed and critical for ORFV detection • Novel and sensitive diagnostic methods are required for ORFV detection.

Identification and molecular characterization of Orf virus infection in occupationally exposed women in South America.

Contagious Ecthyma (CE) is a severe exanthematous dermatitis caused by the Orf virus (ORFV) that mainly affects domestic small ruminants such as sheep and goats. It is a worldwide-distributed occupational zoonosis, particularly infecting those in close contact with animals or animal products such as shepherds, farmers and veterinarians, among others. In the present work, we report the first human CE case confirmed in Argentina. A phylogenetic analysis based on four gene sequences of the isolated strain responsible for the disease showed that this isolate grouped with other ORFV sequences that caused reported CE cases in sheep from the same Argentine province. We also sequenced a sample from a Chilean human case reported in 2017, whose phylogenetic analysis showed that it groups together with other Argentine isolates from locations close to the border with Chile.

Identification, molecular characterization, and pathological features of orf virus in sheep and goats in Punjab province, Pakistan.

Orf virus (ORFV) causes an acute, contagious, skin disease of sheep and goats which is economically important. The objectives of this study were to identify ORFV and to explore its pathological and phylogenetic profiles in 350 goats and 91 sheep of 14 districts of Punjab, Pakistan, from July 2020 to July 2021. Skin scrapings (total no. of samples = 441) of suspected animals were subjected to polymerase chain reactions, phylogenetic analysis, and pathological observations. The partial length of GIF/IL-2 gene (408 bp) was successfully amplified in 58/441 samples. Phylogenetic analysis of GIF/IL2 gene showed that the study isolates belonged to ORFV-cluster I, together with the viruses reported in India and China. Pakistan ORFV isolates were shared 97.6-98.7% nucleotide and 97.6-100% amino acid identities with the reference strain (NC_005336). Moreover, Chinese ORFV-isolates were detected unique multiple amino acid substitutions (F11L, Q21H, D27N, I46V, N49S, N82D, D103N, S129G) with study isolates. Naturally infected animals were anorexic, emaciated, dull, and depressed. The macroscopic lesions included multifocal to coalescing, ulceration followed by proliferative papules, pustules, and crust formation on the epidermis of gums, lips, mouth commissure, muzzles, nose, and udder. Histopathological examination revealed hyperplasia, anastomosing rete ridges formation and degenerative changes, including spongiosis and vacuolation of epidermal cells. Keratinocytes exhibited eosinophilic intracytoplasmic inclusion bodies with pyknotic and karyorrhexis nuclei. This is the first report on molecular characterization of ORFV from Pakistan, with insight into its pathogenesis and comparative analysis of pathological alterations and genetic diversity between ORFV strains reported in different geographical areas.

A survey of contagious ecthyma and molecular characterization of Orf virus in sheep and goats in Nigeria (2014-2016).

Background: Outbreaks of contagious ecthyma (CE) are frequently reported in sheep and goat flocks in Nigeria with severe clinical outcomes. CE is a debilitating and economically important disease primarily affecting sheep and goats caused by the Orf virus (ORFV). Despite field reports of CE in the country, there is no concise country-wide epidemiological data on the disease and limited genetic data of circulating Nigerian ORFV are available in the public domain. Aim: An epidemiological survey of CE and molecular characterization of ORFV circulating in Nigeria from 2014 to 2016. Method: Data were collected using designed questionnaires, administered to veterinarians and farmers in selected States of Nigeria. Samples were collected during passive surveillance for CE from 2014 to 2016 which were analyzed by polymerase chain reaction (PCR). The A32L and B2L genes of circulating ORFV were also characterized. Results: Analysis of the questionnaire showed that 69.54% (n = 82/118) of the farmers claimed to have experienced CE in their flocks with average morbidity and mortality rates of 25% and 15%, respectively. A total of 113 veterinarians participated in the study, with 69.9% (n = 79) familiar with CE and claimed CE causes morbidity rates of 25%-37% and mortality rates of 10%-15% in sheep and goats. Laboratory results revealed that ORFV was detected in 72% (18/25) of outbreak samples analyzed by real-time PCR. Phylogenetic analysis of A32L and B2L genes revealed that Nigerian ORFV sequences belong to clusters I and II and are similar to viruses from India, Ethiopia, and China. Conclusions: This study is the first nationwide epidemiological data on the status of CE in sheep and goats in Nigeria. It is also the first report of molecular characterization of two genes of ORFV circulating and causing outbreaks in small ruminants in the country. This study showed that CE is under-reported, widespread and of economic importance to sheep and goat farmers in Nigeria.

Molecular detection and characterization of Orf virus from goats in Egypt.

Background: Orf is a highly contagious viral skin disease in sheep and goats caused by Orf virus (ORFV) in the genus Parapoxvirus. Although sheep and goats are considered an essential food resource, particularly in Africa, ORFV infection represents an increasing challenge to animal productivity causing high economic losses. Aim: This study aimed to detect and characterize the ORFV in suspected clinically diseased goats in two neighboring Egyptian governorates, Al-Sharkia and Ismailia, flocks during April 2020 and July 2021by using PCR and phylogenetic analysis of partial B2L sequence.he present study indicate the necessity for establishing normal heart values in conscious and anaesthetized individuals. Methods: Kids from two Egyptian governorates showed the clinical picture of ORFV infection. Samples were collected (n = 15) from two different flocks during April 2020 and July 2021. PCR was carried out to detect the ORFV by targeting a highly conserved sequence within ORFV (B2L) gene. To determine the phylogenetic relationship with other ORFV strains, sequencing and phylogenetic analysis were performed. Results: ORFV infection was confirmed in 12 samples of oral scabs (80%) by PCR targeting a highly conserved sequence within B2L gene. Sequencing of DNA products was performed and obtained sequences revealed 100% identity at the nucleotide level. Two ORFVs, one from each outbreak showed 98.2% nucleotide identity with a previous Egyptian ORFV (KP984529) whereas our isolates showed higher nucleotide identities, 99.1% and 98.7% with ORFV strains from neighboring countries, Sudan and Ethiopia, respectively. The phylogenetic tree grouped isolates into two main clusters, cluster I included isolates of this study and foreign ones mainly from China, India, and Sudan. Interestingly, the vaccine strains of ORF used in different countries were grouped in cluster II with previous Egyptian isolate (KP984529), Ethiopian and Israeli ORFV isolates. Conclusion: Molecular characterization of B2L gene of ORFV isolates revealed higher sequence identities and more close genetic relationships with other ORFV strains circulating in neighboring countries than with the Egyptian isolates. These findings provide an insight into the genetic diversity of field ORFV isolates circulating in goats in the Egyptian governorates.

Recombination drives the emergence of orf virus diversity: evidence from the first complete genome sequence of an Indian orf virus isolate and comparative genomic analysis.

Contagious pustular dermatitis is a disease that primarily infects small ruminants and possesses zoonotic potential. It is caused by orf virus (ORFV), a member of the genus Parapoxvirus. In this study, we evaluated an ORFV outbreak in goats in Madhya Pradesh, a state in central India, during 2017. The transboundary potential of this virus was evaluated by constructing phylogenetic trees. The complete genome sequence of an ORFV isolate named Ind/MP/17 was found to be 139,807 bp in length with 63.7% GC content and 132 open reading frames (ORFs) flanked by 3,910-bp inverted terminal repeats (ITRs). An investigation into evolutionary parameters such as selection pressure (θ = dN/dS) and nucleotide diversity (π) demonstrated that ORFV has undergone purifying selection. A total of 40 recombination events were identified, 21 of which were evident in the Ind/MP/17 genome, indicating its ability to generate new variants.

Contagious Ecthyma Dermatitis as a Portal of Entry for Erysipelothrix rhusiopathiae in Muskoxen (Ovibos moschatus) of the Canadian Arctic.

Erysipelothrix rhusiopathiae was detected immunohistochemically in contagious ecthyma (orf virus) dermatitis in two muskoxen (Ovibos moschatus), harvested and found dead in 2014 and 2015, respectively, on Victoria Island, Canada. This may help target further research on E. rhusiopathiae epidemiology and mechanisms of infection in muskoxen, recently associated with widespread mortalities in Canada's Arctic.

Detection of contagious ovine ecthyma (orf) and risk factors for infection in small ruminants in Iran.

The primary cause of contagious ecthyma is the orf virus, the parapoxvirus prototype. It is a viral problem observed in goat and sheep flocks in Iran, causing economic loss. Orf is a zoonosis with little epidemiological investigation present in Iran. The current research aims at determining the status of this virus, and a PCR was used as a confirmatory instrument. We sampled 668 goats and sheep and various breeding systems. Besides, the orf prevalence was studied, and vaccination efficacy was determined. Moreover, the potential risk factors surveyed for infection with ecthyma were identified. Samples were taken from goat and sheep flocks in the present cross-sectional research, and PCR was used for testing orf DNA. A checklist including animals' general information was completed. Data were analyzed using univariate tests (chi-square and t-tests) and multivariable binary logistic regression analysis. Three hundred one (45%) goats and sheep detected orf DNA. The age of 70% of positive cases was below one month. Ecthyma infection was significantly higher in imported breeds (87.3%) than indigenous (39.3%). Ninety-six percent of infected goats and sheep in the present work were not vaccinated against ecthyma. The high prevalence of the orf virus was confirmed among goat and sheep flocks in Iran. It is necessary to train ranchers regarding sanitary actions, quarantine, and application of orf vaccination plans.

The genetic identification of camel contagious ecthyma virus as the causative agent of contagious ecthyma in dromedary camels (Camelus dromedarius) in Qatar.

Camel contagious ecthyma is a contagious viral disease of camels caused by either Orf virus (ORFV) or camel contagious ecthyma virus (CCEV). It has been previously reported and shown to cause economic losses in some camel-rearing countries in Asia and Africa, but has not been detected in Qatar. The purpose of this study was to identify and genetically characterize the contagious ecthyma causative agent in Qatari dromedary camels between 2017 and 2018. Accordingly, we made diagnoses of camel contagious ecthyma based on the clinical signs and genetic analysis of the entire major envelop protein (B2L) gene. The sequence analysis showed that CCEV was the infecting virus, and the B2L gene sequences were highly conserved between the locally infected camels with 100% similarity with isolates from Bahrain. This is the first study reporting the detection of CCEV in Qatar. We suggest that sequencing of the CCEV genome is necessary to determine the origin and relationship of this virus with other members of the parapoxvirus genus.

Circulation of orf viruses containing the NZ7-like vascular endothelial growth factor (VEGF-E) gene type in India.

Orf, a poxviral skin infection of small ruminants is caused by orf virus (ORFV) of the genus Parapoxvirus of the Poxviridae family. Vascular endothelial growth factor (VEGF) is an important virulence factor that is responsible for proliferative lesions in parapoxviral infections. VEGF gene shows high intra- and inter-species variability. Two variants of VEGF have been described globally in ORFV, viz. NZ2- and NZ7-like. In the present study, ORFV isolates of different geographic regions of India were analysed on the basis of the VEGF gene. Indian ORFV isolates showed 95.7-100 % nucleotide (nt) and 78.4-99.3 % amino acid (aa) identity with each other, except ORFV-Assam/LK/14 and ORFV-Meghalaya/03 which shared 85.1-88.35 % and 79.1-81.8 % identity, at nt and aa levels, respectively with other Indian ORFV isolates. All Indian ORFVs under the study demonstrated 83.5-99.1 % nt and 80.5-97.9 % aa identity with NZ7-like VEGF as compared to 41.2-44.8 % nt and 30.7-38.4 % aa identity with NZ2-like VEGF on comparison with global ORFV strains. Phylogenetic analysis based on the VEGF gene showed two clusters of ORFV in which the Indian ORFVs clustered with NZ7-like VEGF from global ORFV strains, mostly from China. Despite the considerable variation, VEGF protein from Indian ORFV strains showed conserved VEGF homology domain with eight cysteine residues. Homology modeling of Indian ORFV strains predicted the presence of extended Loop 3 similar to NZ7-like VEGF. Therefore, the present study showed the circulation of ORFV strains with comparatively less variable NZ7-like VEGF in India which implicates its importance in the epidemiology of ORFV infections in the country.

Human Infection with Orf Virus and Description of Its Whole Genome, France, 2017.

Zoonotic transmission of parapoxvirus from animals to humans has been reported; clinical manifestations are skin lesions on the fingers and hands after contact with infected animals. We report a human infection clinically suspected as being ecthyma contagiosum. The patient, a 65-year-old woman, had 3 nodules on her hands. She reported contact with a sheep during the Aïd-el-Fitr festival in France during 2017. We isolated the parapoxvirus orf virus from these nodules by using a nonconventional cell and sequenced the orf genome. We identified a novel orf virus genome and compared it with genomes of other orf viruses. More research is needed on the genus Parapoxvirus to understand worldwide distribution of and infection by orf virus, especially transmission between goats and sheep.

An association of Orf virus infection among sheep and goats with herd health programme in Terengganu state, eastern region of the peninsular Malaysia.

BACKGROUND: Orf virus causes a scabby skin lesions which decreases productivity in small ruminants. The unknown status of this disease in the eastern region of Peninsular Malaysia warrants a study to determine sero-prevalence of orf with regards to farmers' compliance level towards the Herd Health Program (HHP) programme. RESULTS: Out of 504 animals, 115 were positive for Orf-virus antibodies. An overall prevalence rate of 22.8% indicated a high prevalence of orf disease in this region. It was observed that 25.1% (92/367) of goats were positive and 16.8% (23/137) of sheep sero-converted for Orf virus antibody. Several factors were measured for their possible association with prevalence of Orf virus infection. The prevalence was higher in LY farm, JC breed, kid and female animals, and in the presence of disease lesion. Chi-square analysis showed a significant association of three risk factors which are species, age and sex of the animals (P < 0.05). Notwithstanding, all other variables showed no significant difference (P > 0.05). Farms surveyed usually practised intensive management system, keeping animals in the shade at all time, due to limited availability of suitable land as a free-range grazing area. An interview with small holder farmers revealed a lack of awareness of the main goals of herd health programme. An overall compliance level of 42.7% was observed for all HHP parameters. Among the 14 main components of HHP modules, animal identification had recorded highest compliance level (84.62%) while milking management recorded the least compliance (- 82.69%). That explained why there was a high sporadic prevalence of Orf infection in this region. CONCLUSION: Good herd health supervision is a rehearsal target to prevent an outbreak and the spread of diseases thus reduces economic losses among farmers. Therefore, a good herd health programme should be in place, in order to prevent and control disease transmission as well as to improve herd immunity.