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P F Nettleton

Publications and source records attributed to P F Nettleton.

79 records · Page 5Linked to original sources

The association of calf serum with the contamination of BHK21 clone 13 suspension cells by a parvovirus serologically related to the minute virus of mice (MVM).

An investigation of persistent cell deaths of BHK21 suspension cells during subculturing resulted in the isolation of a viral agent. The agent was isolated from samples of dead cells, cell growth media and 2 batches of calf serum. It was established that the agent was associated with the use of certain batches of calf serum. The isolated virus was found to replicate effectively only in rapidly growing BHK cell cultures. In monolayers it caused the formation of large intranuclear inclusion bodies. The isolate was a strong haemagglutinin; it was stable to ether, chloroform, pH 3 and heating at 56 degrees C. It was shown to be a DNA-virus and by electron-microscopy it was evident as unenveloped spherical, small particles (21 nm in diameter). In sucrose density gradients the virus sedimented as 2 peaks at approximately 114S and 85-92S, and by caesium chloride equilibrium centrifugation, two main peaks at densities of 1.39 and 1.31 g/ml were evident with a minor peak at 1.35. The capsid of the complete virion consisted of 3 polypeptides. The agent has therefore been provisionally designated a member of the Parvoviridae family, genus parvovirus. Serologically it was found to be related to MVM. The isolated parvovirus was inactivated by 0.05 per cent peracetic acid and 0.05 per cent acetylethyleneimine.

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Ruminant pestiviruses.

The ruminant pestiviruses, bovine virus diarrhoea virus (BVDV) and border disease virus (BDV) are highly successful and important pathogens which infect ruminant species worldwide. Although the serological relationships among ruminant pestiviruses require further clarification, there is growing evidence for two antigenic groups, one of which predominates in cattle and one in sheep. The success of pestiviruses stems from the ability of the non-cytopathic (NCP) biotype of the virus to cross the placenta and establish a persistent infection (PI) in the developing foetus. This biotype should be regarded as the 'normal' biotype with the cytopathic (CP) biotype being an abnormal virus that is usually isolated only from PI animals dying from mucosal disease. Recent molecular evidence points to CP viruses arising from their NCP counterparts by recombination events that include the insertion of host RNA and/or the duplication of viral RNA sequences. However, the biological mechanism through which CP viruses kill cells remains unknown. Virtually all CP and NCP viruses cause only mild, transient clinical symptoms in healthy adult animals and stimulate a protective immune response. Despite the urgent requirement for a safe, effective vaccine, there is still no commercial vaccine that has been shown to immunize dams so that foetal infection is prevented. In the absence of an effective vaccine, reliable diagnostic techniques are essential to implement effective control measures. There is now a range of monoclonal antibody-based enzyme-linked immunosorbent assays for identifying PI or convalescent animals. These tests are specific, rapid, sensitive and reliable but may themselves become redundant as they are superceded by ever-increasing molecular biology-based techniques.

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Border disease of sheep and goats.

Border disease (BD) is a congenital virus disease of sheep and goats first reported in 1959 from the border region of England and Wales. BD virus (BDV) is a pestivirus in the genus Flaviviridae and is closely related to classical swine fever virus and bovine virus diarrhoea virus (BVDV). Nearly all isolates of BDV are non-cytopathogenic (ncp) in cell culture. There are no defined serotypes but pestiviruses isolated from sheep exhibit considerable antigenic diversity and three distinct antigenic groups have been identified. Distribution of the virus is worldwide. Prevalence rates vary in sheep from 5 to 50% between countries and from region-to-region within countries. The disease in goats is rare and characterized by abortion. Clinical signs in sheep include barren ewes, abortions, stillbirths and the birth of small weak lambs. Affected lambs can show tremor, abnormal body conformation and hairy fleeces (so-called 'hairy-shaker' or 'fuzzy' lambs). Vertical transmission plays an important role in the epidemiology of the disease. Infection of fetuses can result in the birth of persistently infected (PI) lambs. These PI lambs are viraemic, antibody negative and constantly excrete virus. The virus spreads from sheep to sheep with PI animals being the most potent source of infection. Apparently healthy PI sheep resulting from congenital infection can be identified by direct detection of viral antigen or viral RNA in leukocytes or by isolation of ncp virus from blood or serum in laboratory cell cultures. Isolation of virus is unreliable in lambs younger than 2 months old that have received colostral antibody. The isolation of virus from tissues of aborted or stillborn lambs is difficult but tissues from PI sheep contain easily detectable levels of virus. To detect the growth of virus in cell cultures it is essential to use an immune-labelling method. Acute infection is usually subclinical and viraemia is transient and difficult to detect. Sheep may also be infected following close contact with cattle excreting the closely related BVDV.

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