PATHOLOGICAL AND MOLECULAR DIAGNOSIS OF SHEEP POX

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Date
2015-08
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SRI VENKATESWARA VETERINARY UNIVERSITY, TIRUPATI - 517 502 (A.P) INDIA.
Abstract
ABSTRACT : Sheep farming is the major source of livelihood for landless poor in India. Pox infection in sheep causes a huge economic losses in the Indian subcontinent in terms of poor hide quality, decreased productivity, abortions and even mortality in lambs. Hence, the present study was undertaken to describe the gross and histopathological changes in affected organs and to study the ultrastructural changes in skin and lung and also to confirm the disease by PCR. Sheep pox is caused by SPPV, a member of the genus Capripoxvirus and family Pox viridae. In the present study a total of 1240 sheep (Nellore Dora breed) of both sex and different age groups from various slaughter houses and field were screened for clinical signs/ lesions of sheep pox and of these, 77 animals were found affected (including both ailing and dead) with an overall incidence of 6.2%. Among 77 affected sheep, detailed necropsy was carried out in 16 animals and the representative tissue samples were collected for further studies. A few skin/scab samples from ailing animals were also collected at random for PCR studies. Clinically, the ailing animals showed rise of body temperature, occulonasal discharges and characteristic cutaneous pox eruptions at different stages of development. The symptoms were more severe in young lambs. At necropsy, gross lesions of sheep pox were predominantly seen on the skin and lungs followed by other internal organs. The skin lesions were characterized by papules, nodules and scabs mostly on wool-less parts of the body. The papules were greyish white, circular, hard that coalesced to form white and firm nodules with ulcerated centres. Some papules were encrusted to form the scab. The affected lungs were edematous and congested with irregular areas of consolidation besides rib impressions on surface. A few macules and numerous papules/nodules were noticed along with a very few pustules. Round and white papules were seen on the tongue, rumen, and reticulum. Liver, kidney, heart and spleen showed small, white and circular necrotic areas. Lymph nodes were enlarged and edematous. In the present study, microscopic lesions were noticed in the skin, lungs and in other internal organs like tongue, rumen, reticulum, liver, kidney, heart and spleen. Sections of skin showed hyperkeratosis, acanthosis, parakeratosis, hydropic changes with microvesicle formation and presence of eosinophilic intracytoplasmic inclusion bodies in epidermal layer. In the dermis folliculitis, vasculitis, presence of SPC’s and infiltration of mononuclear cells and fibroblasts were seen. In the lungs, congestion, focal areas of necrosis, proliferative alveolitis, foetalization, thickened interalveolar septa with infiltration of mononuclear cells were seen. The alveolar lumen revealed eosinophilic amorphous material and presence of sheep pox cells with eosinophilic intracytoplasmic inclusion bodies. Proliferative bronchiolitis and peribronchiolar lymphoid aggregations were the other changes observed in affected sheep. The sections of tongue revealed congestion, ulcerations and formation of microvesicles. In the rumen, hyperkeratization, necrosis of squamous epithelial cells and presence of characteristic cytoplasmic inclusions in the prickle cells were seen. Reticulum revealed necrosis, sloughing of squamous epithelium with presence of eosinophilic intracytoplasmic inclusion bodies. Sections of liver showed sinusoidal dilatation, presence of inclusion bodies in few hepatocytes and formation of new bile ductules. The kidney sections showed interstitial nephritis, presence of hyaline casts, eosinophilic intracytoplasmic inclusion bodies in the tubular epithelium and necrotic foci. Sections of heart revealed haemorrhages and infiltration of lymphocytes in between the muscle fibers. Depletion of lymphocytes in the lymphoid follicles was seen in the spleen and lymph nodes of pox affected sheep. In the present study, the ultrastructural changes were studied in the skin and lung tissue samples by TEM. The skin revealed hydropic changes and presence of numerous electron dense particles suggestive of immature virions. In the lung, proliferation of type II pneumocytes, presence of the immature virus particles in affected alveolar epithelial cells and macrophages, thickened interalveolar septa along with nuclear and chromatin changes were seen. The virus suspension from skin papule/ nodule was passaged thrice in embryonated eggs through CAM route that revealed death of embryo as early as 48 hours PI. Grossly, congestion, edema, hemorrhages and necrosis and microscopically congestion were noticed in the CAM. In the present study, molecular diagnosis of pox by PCR was carried out with skin and lung samples from affected sheep using B68 and B69 primers specific for P32 gene of Capripoxvirus that yielded an amplified product of 390 bp, confirming the Capripoxvirus infection in the samples tested. The present study revealed the gross, histopathological and ultrastructural lesions in different organs in natural cases of sheep pox. Further, the disease was confirmed based on the pathological and molecular diagnosis of Capripox infection in the affected sheep in the study area.
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SHEEP ; Pox infection; SPPV; MOLECULAR DIAGNOSIS
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