P‐33 Poxvirus infection in two cats

A 3‐year‐old Burmese cat was presented with a history of nonresolving crusted and papular lesions of the face and prior treatment with prednisolone. Skin biopsies revealed typical pox lesions with hyperplasia and ulceration of the epidermis and eosinophilic cytoplasmic inclusion bodies in the epidermal cells. In the upper dermis there was prominent diffuse mast cell infiltration and mild neutrophilic and eosinophilic inflammation. Rare cytoplasmic inclusion bodies were also present in swollen endothelial cells of dermal venules, which showed no other degenerative changes. Histological diagnosis was confirmed by electron microscopic evidence of pox virus particles in inclusion bodies of epidermal cells. The lesions resolved within 6 weeks with systemic antibiotic therapy and supportive care. A 2‐year‐old domestic short‐haired cat was presented with multiple disseminated papular and ulcerative pox lesions with central eschar over the entire body. Histologically, large epidermal inclusion bodies (up to 6 μm in diameter) were present. Widespread haemorrhage and vascular wall necrosis was visible in the dermis and subcutis. Some subcutaneous vessels showed neutrophilic vasculitis. In addition to diffuse dermal neutrophilic and eosinophilic inflammation, a lymphohistiocytic panniculitis was also present. The cat died as a result of massive haemorrhage and lymphedema, despite supportive care. Funding: Self‐funded.     

Identification of Cytokeratins in Bovine Sperm Outer Dense Fibre Fractions

Outer dense fibres (ODF) are important substructures of mammalian sperm tails that are involved in the regulation of sperm motility. In this study, we investigated the identity of several sodium dodecyl sulphate (SDS)‐insoluble ODF proteins. Bovine ODF were purified by separating sperm heads and tails using ultrasound and Percoll^® density gradient centrifugation. Sperm flagella were treated with the detergent cetyltrimethylammonium bromide (CTAB). CTAB‐insoluble material, which reportedly represents the ODF fraction, was collected, and electron microscopy confirmed a highly purified ODF fraction. We found after solubilization of this fraction with SDS that high amounts of insoluble material were retained after centrifugation. SDS‐insoluble material was collected and quantitatively dissolved in 8 M urea. SDS‐gel electrophoresis in the presence of urea revealed polypeptides with apparent molecular masses of approximately 25, 43, and 50 kDa. Subsequent immunoblotting with anti‐cytokeratin antibodies detected two urea‐soluble, SDS‐insoluble proteins with apparent molecular masses of 45 and 66 kDa. The 45‐kDa protein was identified as cytokeratin 19. An antibody reacting with a palette of cytokeratins (CK 1–18 and CK 20), KL1, was the only antibody that reacted with the 66‐kDa polypeptide. We conclude that sperm ODF fractions contain at least one each of type I and type II intermediate filaments. As keratins and intermediate filaments are described as rope‐like structures, we suggest that these intermediate filaments play an important structural or tension‐bearing role in sperm flagella.