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Biomedical subjects

H W Reid

Publications and source records attributed to H W Reid.

At least 37 records · Page 2Linked to original sources

[Malignant catarrhal fever in zoo ruminants].

An outbreak of malignant catarrhal fever (MCF) in Indian gauer (Bos gaurus gaurus) and Javan banteng (Bos javanicus javanicus) occurred in the year 1964 and killed almost all animals of the groups of these species kept at the Munich zoo at that time. In the consecutive years at highly irregular intervals cases of the head-and-eye-form of MCF occurred sporadically at the zoo in European and American bison (Bison bonasus, B. bison' bison, B. bison athabascae), elk (Alces alces), red deer (Cervus elaphus), Père David's deer (Elaphurus davidianus) and again in gaur and banteng. The clinical and pathomorphological observations, including those of prophylactic and tentative treatment are reported. The subspecies of caprinae and alcelaphinae potentionally latently infected with viral agents of MCF kept at the zoo over the reported period are listed. Some details concerning housing, taking care of the animals etc. of the latently infected carriers of the family caprinae and the susceptible species of bovidae and cervidae are also given as far as they are of epidemiological interest. The results of the retrospective analysis and the results of testing paraffin-embedded tissue samples for the presence of ovine herpesvirus-2 (OHV-2)-specific DNA and alcelaphine herpesvirus-1 (AHV-1)-specific DNA from 1964 up to 1997 are discussed. The negative results for OHV-2-specific DNA suggest that MCF at Munich zoo until 1979 could have been "wildebeest-associated", also because until 1974 blue wildebeest (Connochaetes taurinus taurinus) and until 1983 white-tailed gnu (Connochaetes gnou) were kept. However, positive results for OHV-2-specific DNA in the tissues collected from 1985 onwards, strongly suggest the "sheep-associated" MCF.

Animals↗

A novel strategy for determining protective antigens of the parapoxvirus, orf virus.

We investigated the feasibility of using vaccinia virus (VAC) recombinants containing large multigene fragments of orf virus DNA to identify protective antigens of orf virus (OV). Sixteen OV strain NZ2 DNA fragments with an average size of 11.4 kb were recombined into VAC strain Lister. Each fragment was mapped relative to OV restriction endonuclease maps but was otherwise uncharacterized. Together the recombinants represent 95% of the OV genome in an overlapping manner. Immunofluorescence showed all 16 constructs expressed products recognized by OV antiserum and radioimmune precipitation with the same antiserum allowed the localization of the major antigens of OV to specific recombinants. These data indicated the approximate genomic locations of the genes encoding the OV major antigens and showed that their expression was authentic rather than resulting from read through from VAC sequences adjacent to the site of recombination. Vaccination of OV-naive sheep with the recombinant library provided protection against a subsequent challenge with virulent OV. These data confirm the feasibility of the proposed strategy.

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Identification of ovine herpesvirus-2 infection in sheep.

A polymerase chain reaction test for the detection of ovine herpesvirus-2 (OHV-2) DNA was used to identify sites of OHV-2 infection in peri-natal lambs and in adult sheep. OHV-2 was detected in the nasal secretions from all lambs within a period of two months following birth. Subsequently, OHV-2 DNA was identified in a number of epithelial tissues including the cornea, turbinates and pharynx. In addition, OHV-2 DNA was detected exclusively in B-lymphocytes from six of ten adult sheep tested. An infection cycle for OHV-2 in sheep is proposed which bears similarities with the gammaherpesviruses Epstein-Barr virus and mouse herpesvirus-68.

Animals↗

Definitive identification of louping ill virus by RT-PCR and sequencing in field populations of Ixodes ricinus on the Lochindorb estate.

Rapid and precise virus detection procedures are an important component of any epizootiological study. An automated one tube reverse transcriptase and nested primer polymerase chain reaction (RT-PCR) followed by nucleotide sequencing of the cDNA product, was used for the rapid detection and identification of louping ill (LI) virus in field caught Ixodes ricinus and compared with a classical isolation method i.e. infectivity in cell culture. The results establish the genetic identity of LI virus on the Lochindorb Estate. There was a high correlation between the results obtained by RT-PCR and infectivity assays. RT-PCR and sequencing proved to be a rapid and accurate system for identifying LI virus in field specimens. Development of this system should improve the capacity to undertake detailed epizootiological studies of LI virus.

Animals↗

Efficacy of acaricidal tags and pour-on as prophylaxis against ticks and louping-ill in red grouse.

This paper examines the efficacy of 10% lambdacyhalothrin-impregnated plastic tags and a deltamethrin pour-on preparation in protecting red grouse chicks from parasitism by ticks and subsequent infection with the louping-ill virus. In 1995, ten red grouse hens (Lagopus lagopus scoticus) in a free-living population in north-east Scotland were fitted with lambdacyhalothrin-impregnated plastic tags, glued to radio transmitters. Chicks of more than 10 days of age from a further ten untreated radio-collared hens were caught and fitted with individual tags to the ptagium. Both treatments significantly reduced tick burdens in the short term. The number of larvae and nymphs on chicks up to 45 days was less under both treatments than on control chicks and tagged chicks had fewer nymphs than chicks from treated hens. Nevertheless, treatments did not reduce viral infection rates nor increase survival to 10 weeks, possibly explained by incomplete treatment of tagged broods and/or direct or indirect mortality due to tags. In 1996 chicks in ten broods from hens with radio transmitters were individually treated at 14 days of age at a rate of 1 mg/kg of chick with a deltamethrin pour-on preparation. This preparation significantly reduced the number of larvae and nymphs on grouse chicks 7-10 days after application below the number on untreated controls. At 20 days from application, however, only larval numbers were lower on treated chicks. Nevertheless louping-ill virus infection prevalences were significantly reduced at 35 days of age and survival of chicks to 10 weeks increased.

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Molecular variation, evolution and geographical distribution of louping ill virus.

Following the demonstration that the tick-borne encephalitis (TBE) subgroup viruses are distributed as a cline across the Northern Hemisphere (Zanotto et al., 1995), we have analyzed the dispersal pattern of louping ill (LI) virus, the most westerly located member in the cline. A total number of 21 LI or LI-related virus E gene sequences have been used for a detailed molecular analysis of the evolution, phylogeny and geographical distribution of LI virus in the British Isles and Ireland. The results show that LI virus is genetically stable in general but minor differences enable its separation into four genetically distinct subtypes (genotypes) with clear geographical correlation, designated Type 1 in Scotland and England, Type 2 in Scotland, Type 3 in Wales and Type 4 in Ireland. These data demonstrate that geographically independent evolution of LI viruses has occurred. The molecular systematics and substitutional parameters analyses combined with the clinal distribution of the TBE virus complex allow the assignment of the origin for both Negishi (NEG) virus and a Norwegian isolate to the British Isles. Moreover, proposals for the classification of LI and LI-like viruses which cause encephalomyelitis in sheep, goat or cattle are presented.

Animals↗

Natural transmission of orf virus from clinically normal ewes to orf-naive sheep.

The apparent natural transmission of orf virus from clinically normal ewes to susceptible sheep was observed during a border disease vaccine experiment. The 14 susceptible sheep were persistently infected with border disease virus and had been reared indoors in isolation from other sheep since birth. Their ages ranged from two to four years and they were housed in two groups; group 1 consisted of four sheep persistently infected with the Moredun strain of border disease virus and group 2 consisted of 10 sheep persistently infected with the Oban strain of the virus. On day 0, six sheep were removed from group 2 and rehoused. To the remaining four sheep in each group were added eight four- to six-year-old pregnant conventionally reared ewes at 48 days gestation. Fourteen days later the four sheep in group 1 were moved to another pen housing eight similar five-year-old pregnant ewes at 48 days' gestation, and the four sheep from group 2 were rehoused with their original stallmates. Twenty-one days later lip lesions typical of orf were first observed on the sheep from both groups and the disease spread to all the sheep persistently infected with border disease virus over the next four weeks. Virological and serological evidence demonstrated that the source of infection for the sheep was almost certainly the conventionally reared ewes, on which no lesions resembling orf were observed at any time during the study.

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Tissue culture-propagated orf virus vaccine protects lambs from orf virus challenge.

Twenty, eight-day-old specific pathogen-free (SPF) lambs were vaccinated by a single scarification approximately 4 cm in length on the inner right thigh with a double-pronged applicator. The titre of live virus in the vaccine was 10(7.2) TCID50/ml and the estimated dose per lamb was 0.04 ml. Three months and six months later 10 of the vaccinated lambs and five age-matched unvaccinated control specific pathogen free lambs were challenged by a single scarification with virulent virus on the inner left thigh in the same way. After the vaccination all 20 lambs developed lesions characteristic of orf virus infection that had largely resolved four weeks later, when they all had reciprocal ELISA antibody titres > or = 3200 that persisted in all but one of them until they were challenged. After the challenge, the development of lesions in the vaccinated and unvaccinated sheep was compared daily for four weeks by means of a clinical scoring system. Both groups of vaccinated lambs had significantly lower (P < 0.01) total clinical scores after challenge at three months and six months than the unvaccinated lambs.

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A monoclonal antibody, 3/22, to rabbit CD11c which induces homotypic T cell aggregation: evidence that ICAM-1 is a ligand for CD11c/CD18.

The rabbit CD11c molecule has been characterized by use of new monoclonal antibody, mAb 3/22. Expression of the p150,95 integrin (CD11c/CD18) has been shown by flow cytometry and immunohistochemistry to be restricted to monocytes, macrophages, dendritic cells and a small population of lymphocytes in peripheral blood. No expression on neutrophils could be demonstrated. Incubation of the newly derived CD8+ T cell line, BJ/873, with mAb 3/22 causes homotypic aggregation, which has been shown to be cell surface event that is not dependent on intracellular signaling or on receptor cross-linking. Inhibition studies show that the ligands responsible for this aggregation are CD11c/CD18 and ICAM-1, both of which are expressed on BJ/873. One other rabbit T cell line, K34, that also expresses p150,95 and ICAM-1, shows a similar aggregation response when stimulated with 3/22. Cell lines that express p150,95 but not ICAM-1 do not aggregate. These observations suggest that ICAM-1 is a ligand for activated p150,95.

Amino Acid Sequence↗

The activation status of ovine CD45R+ and CD45R- efferent lymph T cells after orf virus reinfection.

The dynamics and activation status of CD4+ and CD8+ T-cells differentially expressing the CD45R (220 kDa) antigen were studied in prefemoral efferent lymph draining the site of cutaneous reinfection with orf virus. CD4+, CD45R+ lymphoblasts preceded CD4+, CD45R- lymphoblasts during the first 48 h after reinfection. Thereafter, the output of both total and blast-transformed CD4+, CD45R- T-cells increased in proportion to the CD4+, CD45R+ cells for the duration of the virus reinfection. Output of CD8+, CD45R+ T-cells exceeded that of the CD8+, CD45R+ cells both before and after reinfection. However, within the lymphoblast population, CD8+, CD45R+ and CD8+, CD45R- T-cells increased and decreased in parallel. CD4+, CD45R- and CD8+, CD45R- T-cells produced interleukin-2, interferon-gamma and granulocyte-macrophage colony-stimulating factor after culture for 24 h without exogenous restimulation, whereas CD4+, CD45R+ T-cells produced only interleukin-2. The results show that although both CD45R+ and CD45R- alpha beta receptor+ T-cell subsets are activated as a consequence of virus reinfection in vivo, it is the CD45R- subset that predominates in the later stages of reinfection and is the principal cellular source of lymphokines in the efferent lymph.

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Characterisation of the lymphoproliferation in rabbits experimentally affected with malignant catarrhal fever.

Malignant catarrhal fever (MCF) in rabbits caused by the three Herpesviruses: alcelaphine herpesvirus-1 (AHV-1), ovine herpesvirus-2 (OHV-2) and hippotragine herpesvirus-1 (HipHV-1) induced hyperplasia of lymphoid tissues and accumulations of mononuclear lymphoid cells in non-lymphoid tissues. However, certain lymph nodes were affected preferentially. The lymphoid cells in non-lymphoid tissues were CD43+ T-cells which showed evidence of in situ multiplication. A more detailed phenotypic analysis of splenocytes and lymph node cells in AHV-1 infected rabbits suggested that the hyperplasia was probably due to the expansion of CD8+ T-cells. On the basis of these data and the observations of other authors, that no or very little viral expression can be detected in lesions of MCF affected animals, we propose that the pathogenesis of MCF results from a dysregulation of a secretory T-cell activator. The variable pathology induced by the three viruses may reflect a quantitative or qualitative differences in this proposed activator.

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Cyclosporin A abrogates the acquired immunity to cutaneous reinfection with the parapoxvirus orf virus.

The effect of cyclosporin A (CsA) on host immunity to cutaneous reinfection with the parapoxvirus orf virus was studied in 6-month-old lambs. In control reinfected animals, clinical lesions and viral replication (measured by the presence of vesicular/pustular lesions and viral antigen) in regenerating epidermal cells were at a maximum on day 4 with resolution by day 9. Lesion histology revealed recruitment of T cells, B cells and dermal dendritic cells (DDC) which increased and decreased in parallel with the clinical course of the reinfection. In animals treated with CsA (25 mg/kg/day) 1 day before and for 8 days after reinfection, more severe clinical lesions and viral replication typical of primary infections were recorded and had not resolved by 28 days following reinfection. During CsA treatment, the recruitment of T cells, B cells and DDC was inhibited. With cessation of CsA treatment there was dramatic recruitment of CD4+ T cells followed by DDC then B cells to the lesion site but rapid onset of acquired immunity was not recorded. Reverse transcription-polymerase chain reaction (RT-PCR) analysis of cytokine mRNAs from lesion biopsies showed individual sheep variations. However, interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) mRNAs were detected in the control reinfected animals on days 3 and/or 9 after reinfection but not on these days in animals undergoing treatment with CsA. In the untreated lambs there was an inexplicable lack of IL-2 and IFN-gamma mRNAs on day 6 after reinfection. Tumour necrosis factor-alpha (TNF-alpha) and vascular endothelial growth factor (VEGF) mRNAs were unaffected by CsA treatment. The data suggest that CsA abrogates acquired immunity to orf virus reinfection by targetting T-cell lymphokine production.

Animals↗

Cytolytic activity and associated serine protease expression by skin and afferent lymph CD8+ T cells during orf virus reinfection.

Following orf virus reinfection in sheep, CD8+ T cells were recruited to the lesion site in the skin during the period of viral replication and appeared in increased numbers in afferent lymph draining the infection site. A proportion of the CD8+ T cells were activated, particularly in the skin, as determined by their expression of the cytolytic cell-associated serine protease, BLT-esterase. This was a useful marker for activated cytolytic cells as there was a good correlation between afferent lymph CD8+ T cell-mediated lysis of autologous orf virus-infected skin cells and their expression of BLT-esterase in vitro. The majority of the cells that expressed BLT-esterase in the skin and lymph were CD8+ T cells and cytolysis in vitro was predominately by the MHC class I antigen presentation pathway. The conclusion is that orf virus reinfection is characterized by limited viral replication and stimulation of an activated CTL response in the skin.

Animals↗

Isolation of a parapoxvirus from a grey seal (Halichoerus grypus).

A grey seal (Halichoerus grypus) developed cutaneous pocks which progressed to involve the skin extensively, necessitating euthanasia. Macroscopically and histologically the lesions resembled previous descriptions of parapoxvirus infections of seals and virus particles were observed in preparations of a scab and a skin lesion. Suspensions of the scab and skin lesion were prepared and inoculated on to monolayer cultures of grey seal kidney cells. After 25 days in culture and three passages, cytopathic effects were observed and parapoxvirus particles were detected by electron microscopy in the supernatant fluid. Both isolates were adapted to cultures of fetal lamb muscle cells and shown to be antigenically related to orf virus.

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An arbovirus cline across the northern hemisphere.

The mode and tempo of arbovirus evolution and dispersal can help to explain the dynamics of pandemics, viral outbreaks, and emerging viruses. By comparing nucleotide and deduced amino acid sequences of their envelope proteins, we describe the continuous distribution of the tick-borne encephalitis (TBE) complex viruses, the most important flaviviruses in Europe, across major geographical areas and the conditions under which mutations occur. The analyses reveal a correlation between the geographical and genetic distances of these viruses. The arthropod host appears to be a key factor for the formation and maintenance of this cline by constraining TBE dispersal and evolution. This is also illustrated by comparisons with mosquito-borne flaviviruses.

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Johne's disease in a herd of farmed red deer.

An outbreak of Johne's disease in a herd of farmed red deer was studied for four years. Serological, histopathological and cultural techniques were used to monitor the progress of the disease, and delayed type hypersensitivity skin tests were also applied. The results of the serological tests showed that they were poor predictors of future clinical cases and did not consistently identify animals harbouring mycobacteria. The histopathological methods provided a sensitive and specific means of confirming the infection. The skin tests had a low sensitivity and the results were poorly correlated with the serological results in seropositive animals. A vaccination policy was instituted which was accompanied by a change in the pattern of disease. Although the histopathological evidence suggested that the infection was still occurring, there was a marked reduction in the incidence of clinical disease. Vaccinated animals showed a good response to the skin test.

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