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C J Howard

Publications and source records attributed to C J Howard.

At least 145 records · Page 8Linked to original sources

Variation in the intracellular polypeptide profiles from different isolates of bovine virus diarrhoea virus.

Variation of the intracellular polypeptides induced in calf testis cells by 5 cloned isolates of bovine virus diarrhoea virus (BVDV) was examined. Three of the isolates were cytopathic (NADL, C 2415 and Pe 515 c) and two were non-cytopathic (C 1226 and Pe 515 nc) in these cells. The isolates Pe 515 c and Pe 515 nc were both isolated from an animal with clinical signs of mucosal disease. In cells infected with NADL, 8 virus specific proteins (vp 1 to vp 8) with molecular weights ranging from 120,000 (vp 1) to 23,000 (vp 8) were detected. Isolates C 2415 and Pe 515 c gave a similar array of polypeptides to NADL, but the 3 cytopathic isolates could be distinguished by the variation in the molecular weights of some of the proteins. The non-cytopathic isolates could also be distinguished from each other by this type of molecular variation; however, one feature that characterised these strains, when compared to the cytopathic isolates, was the absence of vp 2. Comparison of the polypeptides induced by Pe 515 c and Pe 515 nc showed that apart from the lack of vp 2 in the Pe 515 nc virus profile, the molecular weights of the other viral proteins were similar. This supports serological evidence that for mucosal disease to occur the pair of cytopathic and non-cytopathic viruses must be closely related. Four of the polypeptides induced by Pe 515 c were shown to be glycoproteins.

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Immune response of cattle to respiratory mycoplasmas.

M. bovis and M. dispar produce very different types of response in the calf lung. The immune response of the host plays a major role in the formation of the lesion induced by M. bovis and this response is predominantly Ig mediated. In contrast M. dispar produces a much milder reaction in the lung which may be related to an ability of the mycoplasma to inhibit the host's immune response. Recovery from infection appears to be primarily mediated by IgG present in the lung and functioning in conjunction with alveolar macrophages. The immunity following vaccination which is observed in experimental studies and field trials is also likely to be mediated by the same mechanism.

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Growth of Mycoplasma bovis in organ cultures of bovine foetal trachea and comparison with Mycoplasma dispar.

Inoculation of tracheal organ cultures from bovine foetuses with Mycoplasma bovis resulted in a loss of cellular structure of the lamina propria, followed 20-22 days later by lifting and detachment of overlying epithelium. The effect was associated with large numbers of M. bovis, identified by immunoperoxidase labelling and electromicroscopy, infiltrating between the epithelial cells and amassing in the lamina propria, especially in the region of the basement membrane of the epithelium. Ciliary activity was undiminished for up to 18 days following inoculation and little or no cytopathic effect on the ciliated epithelium was seen in spite of the close proximity of large numbers of organisms. In contrast, M. dispar was restricted to the margin of the ciliated epithelium where, as previously reported, it caused pyknosis, sloughing and flattening of the epithelium with consequent loss of ciliary activity. The cytopathology observed for each mycoplasma bore a close similarity to the behaviour of the two mycoplasmas in vivo and it is suggested that the organ culture system may be a useful and relevant system to elucidate the pathogenic mechanisms for each mycoplasma.

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Comparison by the neutralisation assay of pairs of non-cytopathogenic and cytopathogenic strains of bovine virus diarrhoea virus isolated from cases of mucosal disease.

Neutralising antibody to non-cytopathogenic and cytopathogenic strains of bovine virus diarrhoea virus (BVDV) was assayed in a microtitre test in which cultures of calf testis cells were stained by the immunoperoxidase method to detect viral replication. Fourteen BVDV strains were compared in cross neutralisation tests with antisera prepared in gnotobiotic calves. Ten of the strains comprised five pairs of non-cytopathogenic and cytopathogenic BVDV. Each pair was isolated from an animal with mucosal disease. All five animals were from five separate outbreaks of the disease. Each pair of strains from the same outbreak was found to be antigenically indistinguishable. In contrast, when the coefficient of antigenic similarity was calculated 11 of 45 comparisons between the pairs and 46 of 91 comparisons between all 14 viruses gave R values that distinguished strains. The observations suggest that an antigenic spectrum within a single related group exists for BVDV strains, rather than distinct serotypes. The findings are also consistent with the suggestion that cytopathogenic strains from natural outbreaks of mucosal disease arise by mutation from non-cytopathogenic virus.

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Comparative pathogenicity of Mycoplasma bovis and Mycoplasma dispar for the respiratory tract of calves.

Mycoplasma bovis and M. dispar are both important in the etiology of calf respiratory disease, but marked differences are evident in the pathogenesis of disease caused by the two species. Both attach to bovine cells but vary in their invasiveness. M. bovis is capable of invading the lung parenchyma from the airways, while M. dispar predominantly infects the ciliated epithelial cells lining the respiratory tract. M. dispar causes ciliostasis in fetal-bovine tracheal organ cultures, whereas M. bovis does not have the same dramatic effect on ciliary activity. The immunological response of the host contributes to the lesions produced by M. bovis. Both T- and B-lymphocytes accumulate in the lung, which becomes a site for the production of antibody. In contrast, M. dispar produces a mild cellular response in the lungs of calves and may have immunosuppressive properties.

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Pathogenesis and epidemiology of bovine virus diarrhoea virus infection of cattle.

An outline of the clinical diseases that arise following BVDV infection is given. Isolates of BVDV can be separated into two forms non-cytopathic and cytopathic depending on their effect on cell cultures. In utero infection of the foetus with non-cytopathic virus may result in a number of syndromes, such as abortions, stillbirths or weak calves. It may also result in the birth of calves with a persistent viraemia and these animals may later develop mucosal disease as a result of superinfection with a "homologous" cytopathic strain of BVDV. Acute infection of seronegative animals in usually mild and subclinical. A chronic disease also occurs and this can be protracted with progressive wastage and diarrhoea. This condition has not yet been well defined but it is suggested that it may be the result of superinfection of a viraemic animal with a "heterologous" cytopathic strain of virus. The maintenance of non-cytopathic virus within the cattle population can be either by the slow spread following acute infections of seronegative animals or, more importantly, by spread from persistently viraemic cattle. The cytopathic virus is usually found in association with cases of mucosal disease and may be maintained in the population only by continually arising, possibly by mutation, from the non-cytopathic virus. It is recommended that persistently viraemic animals are eliminated from the herd to avoid in utero infections and the possibility of subsequent mucosal disease.

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Systemic and local immune responses of gnotobiotic calves to respiratory infection with Mycoplasma bovis.

Gnotobiotic calves were inoculated by the intratracheal route with Mycoplasma bovis and the specific antibody response in sera and tracheo-bronchial washings examined by radioimmunoassay. In sera an IgM response which reached a peak two weeks post infection was followed by IgG1 and IgG2 antibody responses. Low levels of IgA antibody were detected in sera three and four weeks after infection. The predominant antibody in tracheo-bronchial washings 2 weeks after infection was IgA. Four weeks after infection IgG1 antibody predominated, but IgG2 and IgA antibodies were also present. Cells containing Ig were present in the cellular accumulations around the necrotic zones produced by M. bovis in the lung parenchyma two and four weeks after infection. IgG1 containing cells predominated in these cellular infiltrates. IgG2 producing cells were the next in frequency. It is concluded that the lung lesion caused by M. bovis is partly due to the host's immune response, presumably contributing to the control of the infection, and that the cells infiltrating the lung are a major source of the local and systemic IgG antibody that is detected after infection. IgA staining cells were observed in the submucosa of tissues from nasal cavity and trachea. These cells are probably the source of IgA in tracheo-bronchial washings and sera since IgA-producing cells were not a predominant component of the lesion in the lung parenchyma.

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Mycoplasma bovis infection in gnotobiotic calves and combined infection with respiratory syncytial virus.

Mycoplasma bovis was inoculated alone or in combination with respiratory syncytial virus into the respiratory tracts of 12 gnotobiotic calves. Clinical signs ranged from transient pyrexia to protracted fever accompanied by severe lower respiratory signs and in one case, arthritis. Pulmonary lesions included foci of coagulative necrosis surrounded by mononuclear cells and suppurative bronchiolitis with varying degrees of lympho-reticular hyperplasia. No enhancement of lesions occurred in the combined infections of M. bovis and respiratory syncytial virus. M. bovis was identified by immunoperoxidase labelling in lesions of necrosis, especially at interfaces between the lesion and mononuclear cells and in bronchiolar exudates. Organisms were also located in necrotic lesions of joint capsules, in tonsillar crypts, and in liver.

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Immune responses to mycoplasma infections of the respiratory tract.

Mycoplasmas are capable of causing respiratory disease in a number of species of animals. The pathogenicity of the mycoplasma species ranges from those that cause major disease outbreaks and economic loss to what might be considered the more highly evolved and successful parasites at the other end of the spectrum that survive for long periods in the host without being recognised and evicted. This prolonged colonisation of mucous membranes which is typical of many mycoplasmas is related to certain unique features of the mycoplasma and its interaction with the hosts immune system. An initial step in infection is the attachment of the mycoplasma to the epithelial lining of the respiratory tract. Lack of cell wall confers plasticity and may engender the intimate association of mycoplasma and host cell that has been noted. This in turn may favour persistence of the extracellular parasite. Before the specific immune response is produced avoidance of the non-specific immune mechanisms would clearly aid survival. Both passive (capsules) and active (toxic effects) mechanisms of avoiding phagocytosis have been proposed. Both humoral and cell mediated responses are generated by mycoplasma infections. The serum antibody response follows the usual course IgM, G and A. The indications of cell mediated immunity that have been reported include; delayed type hypersensitivity reactions, lymphocyte transformation responses and inhibition of macrophage migration. The concept that the pathological lesions are in a large part due to host reactivity is well accepted. The lung lesions may contain infiltrating and dividing lymphocytes some of which are producing specific antibody. Evidence for the lung lesion in some animals being partly due to the hosts cell mediated response has also been produced. The local immune response appears to be of greater relevance to immunity to infection than the systemic response, in general the association between local antibody and immunity is much better than for serum antibody. Of particular note is the high contribution of local IgG production, particularly in the lower respiratory tract. Attempts are now being made to use this increased understanding to produce effective killed vaccines that produce immune responses in the lung. Such studies will hopefully lead to the development of 'killed' vaccines that are effective. It can be urged that mycoplasmas would be less pathogenic if they did not produce an inflammatory response and some species have been shown to have an immunosuppressive effect. Such a property could affect the lesion, and hence pathogenicity, and also aid mycoplasma persistence.(ABSTRACT TRUNCATED AT 400 WORDS)

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An enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to bovine viral diarrhoea virus (BVDV) in cattle sera.

A microtitre ELISA has been established for the quantitation of antibodies to bovine viral diarrhoea virus (BVDV). Single dilutions of sera were assayed and units of antibody were calculated from a standard curve. In order to detect the maximum number of responding animals both IgG1 and IgG2 antibody should be assayed, although detection of IgG1 alone was nearly as effective. The ELISA was as sensitive as the virus neutralization test for detection of antibody; comparison of an ELISA that detected IgG1 plus IgG2 antibody to BVDV with the virus neutralization test gave a correlation coefficient (r) of 0.89 (P less than 0.001 for 95 compared sera). Although similar amounts of IgG1 and IgG2 antibodies were present in sera from both experimentally- and naturally-infected cattle, antibody to BVDV in colostrum and in the sera from young calves was predominantly IgG1. The number of adult cows with antibody was 40 out of 41 while 36 of 44 calves reared in a beef unit were found to have produced antibody by the time they were 31.5 weeks old, an indication of the high prevalence of BVDV in the cattle population.

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Experimental production of fatal mucosal disease in cattle.

Three outbreaks of mucosal disease were investigated. Careful examination of 47 cattle that were persistently viraemic with bovine virus diarrhoea virus (BVDV) revealed no clinical disease, no or low levels of BVDV antibody and only non-cytopathic virus in their blood. The four animals with mucosal disease all showed clinical disease and both cytopathic and non-cytopathic virus in their blood. Following post mortem examination, there were particularly high levels of cytopathic virus in gut tissue. A hypothesis for the induction of mucosal disease is suggested. It states that animals become persistently infected with non-cytopathic virus following in utero infection and when, in post natal life, they become superinfected with a cytopathic virus, then mucosal disease ensues. The experimental reproduction of mucosal disease in support of this hypothesis is described.

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Comparison of bovine IgG1, IgG2 and IgM for ability to promote killing of Mycoplasma bovis by bovine alveolar macrophages and neutrophils.

The IgG1, IgG2 and IgM fractions were purified by chromatography from bovine antisera to Mycoplasma bovis. They were assayed for specific antibody and compared for ability to promote killing of M. bovis by bovine peripheral-blood neutrophils and alveolar macrophages. None of the Ig preparations killed the mycoplasma in the absence of the cells. The IgG1 and IgG2 preparations both promoted mycoplasma killing by the macrophages; IgM appeared to have no effect. The IgG2 preparation promoted killing by the neutrophils but neither the IgG1 or IgM fraction appeared effective.

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Animal ureaplasmas: their ecological niche and role in disease.

Each animal species is colonized by a restricted and characteristic group of ureaplasmas, although it can be demonstrated experimentally that host specificity is not absolute. Ureaplasmas are likely to be complicating agents in several diseases, including respiratory infections, conjunctivitis and urogenital infections; alone they can induce mild disease in the lung and conjunctiva. Ureaplasmas are commonly isolated from the urogenital tract where they have been shown to cause vulvitis. In general, attempts to relate disease to specific serotypes have not been successful.

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Immune response of calves following the inoculation of Mycoplasma dispar and Mycoplasma bovis.

A small but significant reduction in the number of Mycoplasma dispar colonising the respiratory tract after intratracheal challenge was observed in gnotobiotic-calves previously inoculated subcutaneously three times with formalin-killed organisms and oil adjuvant. Injection of M. dispar by the intramuscular route, with oil adjuvant, and 2 weeks later by the intratracheal route, without adjuvant, failed to induce immunity to subsequent intratracheal challenge. Following the subcutaneous injection of killed M. dispar, the amount of antibody detected by single radial haemolysis (SRH) increased markedly with increasing age in groups of calves with average ages of 16 to 155 days when first injected. Most calves aged less than 40 days failed to produce an antibody response to a single injection of M. dispar. With M. bovis a smaller difference was observed between antibody levels generated in calves of different ages; also, calves less than 40 days old produced a detectable SRH antibody response following a single injection of killed M. bovis. IgG1 and IgG2 antibody to M. dispar and M. bovis were measured by ELISA. IgG1 appeared before IgG2 antibody and this was particularly pronounced in younger calves. Also, for both mycoplasmas IgG2 antibody levels were lower in younger than older calves. The IgG1 response to M. dispar was compared in three groups of calves with average ages of 16, 55 and 155 days and was greatest in the oldest and least in the youngest animals. In contrast, the IgG1 response to M. bovis varied little in calves of different ages. It therefore appears that the immune response of young calves to M. dispar is impaired or defective.

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Comparison of ureaplasmas from sheep and goats with Ureaplasma diversum and U. urealyticum.

Ureaplasmas isolated from sheep and goats were compared by immunofluorescence with antisera prepared in calves and by PAGE of polypeptides labelled by growth in the presence of [35S]methionine. The ovine and caprine strains constituted two groups defined by serology and polypeptide composition that were not related to the animal species from which they originated. Strains representing these two groups were compared with Ureaplasma urealyticum (human isolates) and U. diversum (bovine isolates). They could not be classified with either but were more similar to the U. diversum strains.

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A search for new microorganisms in calf pneumonia by the inoculation of gnotobiotic calves.

In a detailed microbiological and pathological study of eight outbreaks of calf pneumonia, which included the inoculation of respiratory material into 18 gnotobiotic calves, a total of five viruses, four species of mycoplasma and 19 species of bacteria were identified. The only microorganism not previously associated with bovine respiratory disease was a coronavirus. The experimental disease produced in the majority of the gnotobiotic calves by the intratracheal inoculation of unpassaged respiratory material, closely resembled the natural disease. However statistical analysis of the results could not ascribe the disease seen to a single microorganism or to a particular combination of microorganisms. Assuming therefore that no microorganisms were missed in the study, it was concluded that the microbiological cause of the experimental disease lay amongst the organisms already found and that one or more of three other factors, discussed in the paper, accounted for the experimental disease produced.

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Identification of ureaplasmas from cattle using antisera prepared in gnotobiotic calves.

Antisera were prepared in gnotobiotic calves to three bovine ureaplasma strains which were selected as representatives of the three clusters of serologically similar, but not identical, strains recognized in previous experiments with rabbit antisera. A further 110 isolates from various organ systems of cattle from six countries reacted by the fluorescent antibody technique with at least one of these bovine antisera and it appears that bovine ureaplasmas can be identified and classified by means of three antisera only. It is suggested that the bovine ureaplasmas could be designated as groups A, B or C, these being further subdivided with rabbit antisera. These findings make possible the screening of cattle for antibody to ureaplasmas with just three antigens and also have implications for immunization.

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