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Characterisation of immunodominant protein encoded by the F1L gene of orf virus strains isolated in Italy.

We analysed the molecular properties of the immunodominant protein of different orf virus strains isolated in Italy. The F1L encoding genes and the deduced amino acid sequences of all strains were determined and compared, and they showed several mutations. Structural analysis was carried out in order to assess the influence of amino acid variations on protein structure demonstrating a conservation of the secondary structure. Western blot analysis and immunogold electron microscopy showed that all orf virus strains were antigenically identical. The results of our study confirmed the immunogenicity of the F1L protein; furthermore, our data suggest a possible involvement of the protein in the virus cycle.

Amino Acid Sequence↗

Causation and epidemiological features of proliferative lip and facial lesions in Lao goats.

Past surveys and studies have identified a syndrome of lip and facial dermatitis as common in Lao goats and identified Orf virus (ORFV) as the likely cause. To more definitively determine the cause of this syndrome, a matched case-control study was implemented in 70 smallholder goat farms and a large-scale goat holding from Savannakhet province, Laos. Forty-one scab samples from goats with lip and facial dermatitis (cases) and 41 paired samples from unaffected animals (controls) were collected from 31 goat holdings over 12&#x2009;months. Samples were tested using quantitative PCR assays targeting possible pathogens causing such lesions. The prevalence of PCR-positive samples in case and control goats was 100% and 95.1%, respectively, for ORFV (P&#x2009;=&#x2009;0.15) and 19.5% and 29.3% (P&#x2009;=&#x2009;0.3) for Dermatophilus congolensis. Pseudocowpox virus and bovine popular stomatitis virus were not detected in any samples. There was a clear difference between the log10 genome copies (GC) per &#xb5;L DNA extract of ORFV in positive samples from cases (7.00&#x2009;&#xb1;&#x2009;0.17) and controls (2.62&#x2009;&#xb1;&#x2009;0.18, P&#x2009;<&#x2009;0.0001). This was not true for D. congolensis with similar GC detected in the case and control goats. More than 4 log higher GC in affected cases identifies ORFV as the likely cause of the syndrome in Lao goats with little role for D. congolensis. Phylogenetic analysis showed no genetic variation of ORFV in the study samples and close similarity to sequences from China. The findings suggest that orf disease has a sporadic distribution, mostly occurring in goats during the first 6-months of life.

Animals↗

Ocular infection with orf virus.

Orf virus infection of the pericanthal eyelid skin and conjunctiva was diagnosed by electron microscopy. Orf is a paravaccinia virus responsible for contagious pustular dermatitis in sheep and goats. It occasionally affects humans. The patient was a farmwife who had recently handled lambs. She had a raised ulcerating lesion at the inner canthus, follicular conjunctivitis, and lymphadenopathy. Treatment with 35% idoxuridine in dimethylsulfoxide on the skin and 0.5% idoxuridine ointment on the conjunctiva led to resolution after three weeks.

Adult↗

Recurrent orf in an immunocompromised host.

A 30-year-old farmer with Nezelof's syndrome developed a giant orf on his hand. Recurrence followed surgical excision. Three excisions and split skin grafts were required before its eradication. He re-presented 8 years later with a further orf on the finger of his opposite hand. This lesion had not grown to the exuberant proportion of the previous lesion, but it defied repeated excisions, and various medical therapies including idoxuridine, interferon and transfer factor. Excision with hypochlorite dressings perioperatively and delayed split skin grafting led to eventual eradication.

Adult↗

Human orf.

Human Orf is an uncommon viral disease acquired through contact with infected sheep and goats. We report five cases of human Orf acquired while preparing mutton. The clinical picture and the management of human Orf are presented. Awareness of the benign nature of the condition is important in preventing ill-advised therapy.

Adult↗

Studies of the pathogenesis of orf virus infection in sheep.

Damage to the skin is essential for the establishment of orf virus infection and the development of typical lesions. However, analysis of the pathogenesis of experimental lesions induced by viral challenge of mildly abraded skin, indicated that the virus does not establish in the damaged epidermis, but replicates in the cells of an underlying replacement epidermal layer derived from the walls of the wool follicles. The skin reaction consists of a cellular response with necrosis and sloughing of the affected epidermis and underlying stratum papillare of the dermis. Healing is then completed by the formation of a third epidermis derived from the deeper portions of the wool follicles. Previous cutaneous infection did not prevent reintroduction of the disease, even on the same area of skin although the lesions were less severe and persisted for a shorter period.

Animals↗

Sero-surveillance for orf antibodies in sheep and goats in Saudi Arabia employing the ELISA technique.

A serological survey was conducted on sheep and goats in Saudi Arabia to detect humoral antibodies against the orf virus. The ELISA technique was superior to the AGID and CFT in detecting such antibodies. Of the 239 abattoir serum samples examined, 60 per cent had orf antibodies by ELISA while 94.8 per cent of the convalescent sera had antibodies detectable by ELISA. With the lack of vaccination and the reports that the clinical disease is commonly seen in different parts of the Kingdom, it may be that orf is becoming enzootic in this country. Recommendations for implementing a satisfactory vaccination regimen are given.

Animals↗

Immune response of lambs to experimental infection with Orf virus.

A group of six specific pathogen free (SPF) lambs were infected epidermally with Orf virus. Seven weeks later they were reinfected. For a period of 4 weeks after each inoculation they were observed clinically and blood was collected for analysis of virus specific antibody measured by ELISA and peripheral blood lymphocyte (PBL) proliferative response to various viral antigens. After the primary infection all animals showed clinical signs of Orf, namely vesicle formation which became pustular followed by scabbing; this steadily became heavier prior to shedding and the resolution of the infection by about 4 weeks. The severity of infection varied within the group. Little lymphoproliferative activity was recorded during the primary infection, although five/six test animals had positive lymphoproliferative responses to an sodium dodecyl sulphate (SDS) solubilised scab purified Orf virus preparation at some point between days 7 and 14 after inoculation. All animals seroconverted to Orf virus, lymphoproliferative activity always preceding specific antibody detection. Resolution of the secondary infection was very rapid. Vesicles were visible by day 2 after inoculation which became pustular followed by scab formation and resolution in the majority of animals by day 8. All animals showed a significant (greater than four-fold) rise in specific antibody titre following secondary inoculation. The proliferative activity of PBL's was much greater than that recorded for the primary infection although the magnitude of this response varied greatly between individuals.

Animals↗

Response of efferent lymph and popliteal lymph node to epidermal infection of sheep with orf virus.

Functional and phenotypic changes in the cell populations were monitored in the popliteal efferent lymph of sheep following experimental epidermal infection with orf virus. In another group of sheep, cells from the popliteal lymph node draining the site of infection were similarly monitored and compared with the cells from contralateral popliteal and mesenteric lymph nodes. All sheep showed serological evidence of previous exposure to orf virus. Following infection, anti-orf antibody titres rose and efferent lymphocyte and blast cell output increased. Interferon-like activity was detected in efferent lymph early after orf virus but not mock infection. Lymphocytes from the draining popliteal lymph node showed antigen-specific lymphoproliferation on Days 3-7 while cells in the efferent lymph demonstrated proliferative activity on Days 4-6. The requirement for exogenous antigen-presenting cells in the culture of efferent lymphocytes varied between individual sheep. The culture supernatant from proliferating lymph node cells contained interferon-like activity but no anti-orf antibodies, the reverse of that from cultured efferent lymphocytes, perhaps indicating a different reactive T cell population. During the course of the experiment there was an increase in the percentage of efferent lymphocytes expressing MHC Class II antigens and surface immunoglobulins, the latter being recorded as a double peak. The short-term nature of the local T cell response may in part explain the incompleteness of immunity to orf virus in sheep.

Animals↗

Human orf and milkers' nodule: a clinicopathologic study.

We report the clinical and histopathologic features of 17 patients with orf or milkers' nodule infection. The majority were male, 12 to 65 years of age, and gave a history of contact with farm animals. Most lesions affected the hands or arms, ranged in size from 1 to 3 cm, and occurred on average 3 weeks after presumed exposure. On low-power examination the epidermis showed endophytic strandlike proliferations and the dermal papillae were distended by intense edema. There was massive capillary proliferation and dilation and a dense inflammatory infiltrate. High-power examination revealed epidermal viral cytopathic changes with inclusion bodies, clumping of keratohyalin, and cytoplasmic vacuolation that had a distinctive "spongiform" appearance within follicular structures. We conclude that orf and milkers' nodule infection have distinctive histopathologic features, and, in contrast to some previous reports, viral changes may frequently be found.

Adolescent↗

A seven-year-old girl with orf of the hand.

A seven-year-old child presented with orf of the hand. There was a clear history of contact with sheep. The lesion was incised and drained and healed spontaneously within five weeks. The features of orf infection are described as a reminder of a diagnosis to be remembered in the urban accident and emergency department or hand clinic.

Animals↗

The response of the supramammary lymph node of the sheep to secondary infection with orf virus.

The response of the supramammary lymph node of seven sheep to secondary infection with orf virus was examined by cannulating the efferent lymphatic before infecting the drainage area of the node. All animals developed typical orf lesions and responded after an initial lag period with an increase in total cell output paralleled by a rising proportion of lymphoblast cells. Most lymphoblast contained immunoglobulin with a predominance of the IgG class. When cultured, these cells produced measurable amounts of virus-specific antibody. The proportion of cells which stained with a T-cell-specific monoclonal antibody was measured in two of the sheep and found to decrease as the response developed. These data suggest that the nodal response is directed mainly towards the production of virus-specific antibody. The extent of T-cell involvement remains unclear.

Animals↗