Previous reports have suggested that T. orientalis group species may be non-pathogenic in healthy cattle, and an incidental finding in blood samples. However, this investigation provided evidence that in New Zealand, this pathogen is capable of causing clinical disease in cattle not necessarily debilitated by another disease. The potential for disease should be considered when naive cattle are brought in from non-endemic to endemic regions, for instance cattle from the South Island moved to regions where the vector for T. orientalis group species, Haemaphysalis longicornis, is active, and T. orientalis is present.
Fresh and formalin-fixed tissues and blood samples in ethylenediaminetetraacetate were collected from cattle, deer and buffalo with clinical signs suggestive of malignant catarrhal fever (MCF). In addition, formalin-fixed, paraffin-embedded tissue blocks collected from these animals and retrospectively from field cases of MCF were examined. DNA samples extracted from these samples were analysed by polymerase chain reaction (PCR) assay using primers specific for the sheep-associated (SA)- and wildebeest-associated (WA)-MCF viruses. Both the SA-MCF virus and WA-MCF virus PCR yielded positive results which were in nearly complete agreement with the histopathological diagnoses of MCF in fresh and formalin-fixed, paraffin-embedded tissue from 29 cattle, 24 deer and three buffaloes. Some blood samples tested by the two assays indicated that some of the infected cattle were possible carriers.
Human papillomaviruses (HPVs) are associated with benign and malignant neoplasms of the cervix. One of the criteria for their etiologic role requires an assessment of whether virtually all or only a small fraction of lesions contain viral genomes. DNA preparations from colposcopically directed punch biopsies of cervical lesions were analyzed by Southern blot hybridization and the polymerase chain reaction (PCR) for the presence of HPV DNA. The biopsy specimens represented different pathologic entities (koilocytosis, condyloma, cervical intraepithelial neoplasia, and invasive carcinoma). In Southern blot hybridization with radioactive probes for HPV 11, 16, 18, 31, and 33, HPV DNA was detected in 74% of the biopsy specimens (42 of 57 cases), with the predominant types being HPV 16 and HPV 18. In contrast, after PCR amplification with primers yielding fragments of characteristic size for HPV 11, 16, and 18, the analysis of the same 57 biopsy specimens revealed that all samples were positive for at least one HPV type. To exclude false-positive PCR results, controls without HPV DNA were interspersed at regular intervals, and results were evaluated only if these controls remained HPV negative. To exclude false-negative results due to failure of the reaction, a target sequence within the c-Ha-ras-1 gene was used as an internal control. All HPV typing results obtained by Southern blot hybridization were in agreement with HPV typing by PCR. The higher number of positive samples in the latter analysis stems from the increased sensitivity of PCR, which was which was effective in identifying as few as 10-100 HPV DNA molecules; in contrast, the sensitivity of Southern blot hybridization was 1 pg, or approximately 10(5) molecules of HPV DNA. The authors conclude that, with sufficiently sensitive diagnostic methods, HPV DNA can be detected in most, if not all, neoplastic cervical lesions.
Chicken embryo fibroblasts (CEFs) and Vero cells infected with infectious bursal disease virus (IBDV) exhibited the biochemical feature of apoptosis. Agarose gel electrophoresis of DNA extracted from IBDV-infected cells revealed the characteristic laddering pattern of DNA fragmentation, which was more intense in infected CEFs than in Vero cells. The appearance of apoptotic nucleosomal DNA fragments in IBDV-infected CEFs was independent of virus replication and occurred at an early stage following in vitro infection.
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