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The Pirbright Institute publication directory contains details of selected publications written by our researchers.

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Beard P M, Daniels M J, Henderson D, Pirie A, Rudge K, Buxton D, Rhind S, Greig A, Hutchings M R, McKendrick I, Stevenson K, Sharp J M (2001)

Paratuberculosis infection of nonruminant wildlife in Scotland

Journal of Clinical Microbiology 39 (4), 1517-1521
Publisher’s version: http://dx.doi.org/10.1128/jcm.39.4.1517-1521.2001

Abstract

Recent reports of natural paratuberculosis (or Johne's disease) in rabbits, foxes, and stoats has focused debate on the presence and importance of wildlife reservoirs in the epidemiology of this disease. This paper describes an extensive study investigating 18 nonruminant wildlife species for evidence of paratuberculosis. Using both culture and histopathological analysis, fox, stoat, weasel, crow, rook, jackdaw, rat, wood mouse, hare, and badger were found to harbor Mycobacterium avium subsp, paratuberculosis, the causative organism of paratuberculosis, suggesting that the epidemiology of this disease is more complex than previously realized.

Beard P M, Rhind S M, Buxton D, Daniels M J, Henderson D, Pirie A, Rudge K, Greig A, Hutchings M R, Stevenson K, Sharp J M (2001)

Natural paratuberculosis infection in rabbits in Scotland

Journal of Comparative Pathology 124 (4), 290-299
Publisher’s version: http://dx.doi.org/10.1053/jcpa.2001.0466

Abstract

Natural paratuberculosis infection of rabbits (Oryctolagus cuniculus) was recently diagnosed in Scotland, and an investigation into the pathology of the disease in wild rabbits is reported in this paper. Evidence of Mycobacterium avium subsp. paratuberculosis (M.a. paratuberculosis) infection was detected in 22% of 110 rabbits: the organism was cultured from 17 of 110 rabbits, Land histopathological lesions consistent with many. paratuberculosis infection were noted in 18 of 98 rabbits examined. No macroscopical lesions suggestive of M.a. paratuberculosis infection were observed. The histopathological le,ions were either severe or mild. Severe lesions consisted of extensive macrophage granulomata and numerous giant cells, with many intracellular acid-fast bacteria in the small intestine. For the examination formalin-fixed, paraffin wax-embedded tissues, neither immunohistochemistry nor the polymerase chain reaction was as sensitive a method of diagnosis as histopathology.

Beard P M, Stevenson K, Pirie A, Rudge K, Buxton D, Rhind S M, Sinclair M C, Wildblood L A, Jones D G, Sharp J M (2001)

Experimental paratuberculosis in calves following inoculation with a rabbit isolate of Mycobacterium avium subsp paratuberculosis

Journal of Clinical Microbiology 39 (9), 3080-3084
Publisher’s version: http://dx.doi.org/10.1128/jcm.39.9.3080-3084.2001

Abstract

The role of wildlife species in the epidemiology of paratuberculosis has been the subject of increased research efforts following the discovery of natural paratuberculosis in free-living rabbits from farms in east Scotland. This paper describes the experimental inoculation of young calves with an isolate of Mycobacterium avium subsp. paratuberculosis recovered from a free-living rabbit. After a 6-month incubation period, all eight calves inoculated with the rabbit isolate had developed histopathological and/or microbiological evidence of M. avium subsp. paratuberculosis infection. Similar results were obtained from a group of calves infected with a bovine isolate of M. avium subsp. paratuberculosis. The virulence of the rabbit isolate for calves demonstrated in this study suggests that rabbits are capable of passing paratuberculosis to domestic ruminants and that wildlife reservoirs of M. avium subsp. paratuberculosis should therefore be considered when formulating control plans for the disease.

Carrasco L, Lima J S, Halfen D C, Salguero F J, Sanchez-Cordon P, Becker G (2001)

Systemic aspergillosis in an oiled Magallanic penguin (Spheniscus magellanicus)

Journal of Veterinary Medicine Series B-Infectious Diseases and Veterinary Public Health 48 (7), 551-554
Publisher’s version: http://dx.doi.org/10.1046/j.1439-0450.2001.00456.x

Abstract

This report describes a case of fatal aspergillosis caused by A. fumigatus during the recovery of an oiled Magallanic penguin. The possible role of aspergillosis as a possible complication responsible for the mortality of penguins surviving the first days of treatment for oil is emphasized.
Charleston B, Fray M D, Baigent S, Carr B V, Morrison W I (2001)

Establishment of persistent infection with non-cytopathic bovine viral diarrhoea virus in cattle is associated with a failure to induce type 1 interferon

Journal of General Virology 82, 1893-1897
Publisher’s version: http://dx.doi.org/10.1099/0022-1317-82-8-1893

Abstract

The establishment of persistent infections with non-cytopathic bovine virus diarrhoea virus (ncpBVDV) is crucial for the maintenance of BVDV in cattle populations. Also, super-infection of persistently infected individuals with antigenically homologous cytopathic BVDV (cpBVDV) results in fatal mucosal disease. Persistent infection with ncpBVDV is established by infection of the foetus during the first trimester of pregnancy. It has been shown previously that foetal infection with cpBVDV does not result in persistent infection. Infection of cells in vitro has demonstrated that cpBVDV induces type I interferon (IFN), whereas ncpBVDV fails to induce IFN. In this study we demonstrate that foetal challenge with cpBVDV results in IFN production, whereas ncpBVDV does not. These findings strongly suggest that the ability of ncpBVDV to inhibit the induction of type I IFN has evolved to enable the virus to establish persistent infection in the early foetus.
Charleston B, Hope J C, Carr B V, Howard C J (2001)

Masking of two in vitro immunological assays for Mycobacterium bovis (BCG) in calves acutely infected with non-cytopathic bovine viral diarrhoea virus

Veterinary Record 149, 481-484
Publisher’s version: http://dx.doi.org/10.1136/vr.149.16.481

Abstract

Acute infection of calves, previously vaccinated with bacille Calmette-Guerin (BCG), with non-cytopathic viral diarrhoea virus (BVDV) resulted in the temporary suppression of two in vitro assays used to monitor Mycobacterium bovis infection. Lymphocyte proliferation and interferon-gamma production by whole blood cultures containing purified protein derivatives prepared from Mycobacterium avium (PPD-A) and M bovis (PPD-B) were markedly suppressed. The implication is that acute infections of cattle with non-cytopathic BVDV may temporarily compromise diagnostic tests for M bovis infections and result in a failure to identify cattle with tuberculosis.
Chernukhin I V, Seago J E, Newbury S F (2001)

Drosophila 5′ → 3′-exoribonuclease Pacman

Methods in Enzymology: Ribonucleases, Pt B edited by A. W. Nicholson 342, 293-302
Publisher’s version: http://dx.doi.org/10.1016/S0076-6879(01)42553-5

Abstract

This chapter concentrates on the methods used to express a Drosophila recombinat 5? ? 3?-exoribonuclease, purify the protein, and analyze its activity in vitro. Analysis of early development in Drosophila has shown that RNA localization, control of translation, and mRNA stability are intimately linked. Generally, translational repression leads to degradation of an RNA, and failure of an RNA to localize correctly also leads to its degradation. Work on the yeast Saccharomyces cerevisiae has identified many ribonucleases and associated factors that control mRNA decay, RNA splicing, and rRNA processing. In yeast, it has been shown that 3? ? 5? degradation/processing of RNA requires the exosome, which is a complex of at least 10 proteins. Degradation of RNA in a 5? ? 3? direction occurs by initial decapping of the mRNA, followed by 5? ? 3? degradation of the RNA by Xrn1p. The two decapping proteins (Dcp1p and Dcp2p) and Xrnlp have been shown to be complexed to seven Lsm proteins, which are likely to form a ring encircling the RNA. To understand the role of RNA stability in development, a number of approaches can be used. Once the genes encoding particular ribonucleases or associated factors have been identified, then expression of the RNA during development can be determined. These techniques have been used to show that the 5? ? 3?-exoribonuclease Pacman is differentially expressed during development.

Fernandez-Fernandez M R, Lucini C, Lopez-Moya J J, Guo H, Martinez-Torrecuadrada J, Casal J I, Mourino M, Plana-Duran J, Rivera J, Rodriguez J F, Montoya M, Del Val M, Garcia J A (2001)

Use of plum pox potyvirus as an expression vector in plants

Molecular Farming. Proceedings of the OECD Workshop held in La Grande Motte, France, 3-6 September, 2000., 161-172
Publisher’s version:

Abstract

Plum pox potyvirus (PPV) is the causal agent of a devastating disease that affects stone fruit trees of the Prunus genus. However, it is also able to infect herbaceous hosts from several different families. This broad host range confers PPV an added value regarding its interest as a putative expression vector in plants. Two strategies have been used to express foreign sequences of interest using vectors based on PPV. The N-terminal region of the capsid protein (CP) has been used to express epitopes on the surface of virion particles. Four different insertion sites were evaluated, but only three of them have rendered viable chimeras. These vectors have shown notable differences in tolerance to sequence insertions. Foreign antigenic peptides expressed in them were easily recognized by specific antibodies. Moreover, sequences cloned at one of these vectors (position 12 of CP) were able to elicit an efficient B-cell response in experimental hosts. Epitope mapping by pepscan analysis and fine tuning of cloning positions are being conducted in an attempt to select optimal sites for epitope insertion. The second approach that we have followed is the use of PPV as an expression vector of whole independent proteins. Two insertion sites have been selected, one between P1 and HC proteins (PPV-XS) and the other between NIb and CP proteins (PPV-NK). A double expression vector has been constructed, which allows production of two foreign proteins with a single vector. ifferent reporter genes have been cloned in both insertion sites. The VP60 structural protein of rabbit hemorrhagic disease virus (RHDV) was also successfully expressed making use of PPV-NK vector. Extracts from the VP60-expressing plants were able to induce a remarkable immune response against RHDV in its natural hosts, rabbits. Moreover, these animals were protected against a lethal challenge with RHDV.
Gomez-Villamandos J C, Ruiz-Villamor E, Bautista M J, Sanchez C P, Sanchez-Cordon P J, Salguero F J, Jover A (2001)

Morphological and immunohistochemical changes in splenic macrophages of pigs infected with classical swine fever

Journal of Comparative Pathology 125 (2-3), 98-109
Publisher’s version: http://dx.doi.org/10.1053/jcpa.2001.0487

Abstract

Classical swine fever (CSF) was induced in 20 pigs by inoculation with a virulent strain of CSF virus to determine sequential changes (2, 4, 7, 10 and 14 days post-inoculation) in the number and morphology of splenic macrophages (red pulp and lymphoid marginal zone) and thus to assess the role of these cells in the pathogenesis of the disease. The first splenic cells to be infected with CSF virus were macrophages in the marginal zone followed by other macrophage populations. The initial phase of CSF was associated with an increase in splenic macrophage numbers in the marginal zone and a decrease in the red pulp. Subsequently, the numbers in the red pulp increased. The study suggested that infection, mobilization and apoptosis of splenic macrophages play an important role in the spread of CSF virus in vivo. Moreover, the secretory changes that occurred in macrophages in the initial phase of the infection suggested that macrophages release chemical mediators capable of modulating pathogenesis, (C) 2001 Harcourt Publishers Ltd.
Graham S P, Brown D J, Vatansever Z, Waddington D, Taylor L H, Nichani A K, Campbell J D, Adamson R E, Glass E J, Spooner R L (2001)

Proinflammatory cytokine expression by Theileria annulata infected cell lines correlates with the pathology they cause in vivo

Vaccine 19 (20-22), 2932-2944
Publisher’s version: http://dx.doi.org/10.1016/S0264-410X(00)00529-6

Abstract

Control of Theileria annulata is currently best achieved by the use of live attenuated cell line vaccines. However, the mechanisms underlying attenuation are unclear and there is a need to rapidly produce new cell line vaccines, which could safely and effectively vaccinate cattle against tropical theileriosis. There is increasing evidence to suggest that proinflammatory cytokines produced by T. annulata infected cells play a central role in both pathology and immune evasion. This study aimed to test this hypothesis and to evaluate cytokine expression as a marker of virulence. The pathogenicity and protective efficacy of cloned T. annulata cell lines that expressed different levels of proinflammatory cytokines were compared. In two independent trials using different stocks of T. annulata, cell lines that expressed higher levels of proinflammatory cytokines induced severe reactions, and in some cases death, when used to vaccinate groups of cattle. In contrast, low cytokine expressing lines induced low post-vaccinal reactions. The results clearly demonstrated that cytokine expression by T. annulata infected cells could be used as a marker of virulence and provided strong evidence to support a role for cytokines in the induction of pathology. Both high and low cytokine expressing cell lines protected cattle against heterologous challenge infection, offering the possibility of using cytokine expression to rapidly select new safe, potent vaccines against tropical theileriosis without the need for culture attenuation.

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