Ninety-one reptiles were examined for the presence of yeasts by standard protocols and pathohistological methods. Yeasts were isolated from 42 of the animals. Representatives of herbivorous families (Testudinidae and Iguanidae) carried yeasts more often than animals belonging to carnivorous taxa (Boidae and Emydidae). Yeasts were most often isolated from the gastrointestinal tract, and in 24-6 per cent of cases they could be cultured from the oral cavity and/or cloaca of living animals. Postmortem examination revealed that the intestines of 80-6 per cent of the animals carried yeasts. In all, 56 isolates, belonging to the genera Candida (32), Trichosporon (11), Torulopsis (9) and Rhodotorula (3), and one perfect yeast were obtained. The species included taxa potentially pathogenic to man. However, no sufficiently reliable criteria could be established to prove that positive culture results were associated with disease. In the reptiles examined postmortem, three cases of dermatomycosis were detected histologically. No case of organ mycosis was identified.
A new class of highly antigenic, MHC-II–restricted mycobacterial lipopeptides that are recognized by CD4-positive T lymphocytes of Mycobacterium tuberculosis–infected humans has recently been described. To investigate the relevance of this novel class of mycobacterial Ags in the context of experimental bacille Calmette-Guérin (BCG) vaccination, Ag-specific T cell responses to mycobacterial lipid and lipopeptide-enriched Ag preparations were analyzed in immunized guinea pigs. Lipid and lipopeptide preparations as well as complex Ag mixtures, such as tuberculin, mycobacterial lysates, and culture supernatants, all induced a similar level of T cell proliferation. The hypothesis that lipopeptide-specific T cells dominate the early BCG-induced T cell response was corroborated in restimulation assays by the observation that Ag-expanded T cells specifically responded to the lipopeptide preparation. A comparative analysis of the responses to Ag preparations from different mycobacterial species revealed that the antigenic lipopeptides are specific for strains of the M. tuberculosis complex. Their intriguing conservation in pathogenic tuberculous bacteria and the fact that these highly immunogenic Ags seem to be actively released during in vitro culture and intracellular infection prompt the urgent question about their role in the fine-tuned interplay between the pathogen and its mammalian host, in particular with regard to BCG vaccination strategies.
The workshop on Three Rs Approaches in the Production and Quality Control of Fish Vaccines aimed a) to identify animal tests currently stipulated for the production and quality control of fish vaccines and to highlight animal welfare concerns associated with these tests; b) to identify viable options to replace, reduce, and refine animal use for fish vaccine testing; and c) to discuss the way forward and set out how the Three Rs may be implemented without jeopardizing the quality of the vaccines. The workshop participants - experts from academia, regulatory authorities, a scientific animal welfare organization, and the fish vaccine industry - agreed that efforts should be undertaken to replace the vaccination-challenge batch potency testing with tests based on antigen quantification or antibody response tests. Regulatory requirements of questionable scientific value and relevance for the quality of fish vaccines, such as the re-testing of batches produced outside Europe, or the double-dose batch safety test, should be re-considered. As an immediate measure the design of the current animal tests should be evaluated and modified in the light of refinement and reduction, for example, the number of unprotected control fish in vaccination-challenge tests should be reduced to the minimum.
This paper reports on the viral content of up to 52 tissue and organ samples of 18 individual large psittacines which died during an epornithic of Pacheco's parrot disease (PPD) caused by psittacid herpesvirus 1 (PsHV1). Associated clinical signs and pathological lesions are described. The large spectrum of samples found to be positive for PsHVl suggests that birds succumb to PPD during viraemia. Tissues and organs from which the virus could be isolated included the integument and associated structures, the muscular, respiratory and circulatory system, bone marrow, the nervous system, thyroid and adrenal glands, spleen and liver, the urogenital tract and the gastro-intestinal tract. Nevertheless, individual and organ (but not species)-specific variation does occur. Virus isolation appears to be most promising from the respiratory, vascular and nervous system and the liver. Highest titres were obtained from heart blood and liver (up to 7.6 log(10)/g tissue), airsac, Nervus vagus and pulp and quill of pin feathers. Pin feathers may therefore be suitable for in-vivo diagnosis. In contrast, HV could not be isolated from any of the feather vanes examined. For the most part, post mortem lesions do not reflect the organ pattern found to be most permissive for virus replication as judged by the success of virus isolation and virus titres. A closer quantitative correlation is indicated for the lungs, spleen and liver, only. Corresponding findings as to frequency of gross pathological lesions and virus quantification appear to be restricted to the liver. In accordance with clinical observations and experimental findings, tissue virus content indicates that horizontal spread of herpesviruses is mediated by cloacal contents or secretions from the respiratory system.
Within the Innovative Medicines Initiative 2 (IMI 2) project VAC2VAC (Vaccine batch to vaccine batch comparison by consistency testing), a workshop has been organised to discuss ways of improving the design of multi-centre validation studies and use the data generated for product-specific validation purposes. Moreover, aspects of validation within the consistency approach context were addressed. This report summarises the discussions and outlines the conclusions and recommendations agreed on by the workshop participants.
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