CD40/CD154 interactions are required for the optimal maturation of skin-derived APCs and the induction of Helminth-specific IFN-? but not IL-4

The mechanisms through which Schistosoma mansoni larvae induce Th1 rather than Th2 immune responses are not well understood. In this study, using CD154–/– mice exposed to radiation-attenuated S. mansoni larvae, we demonstrate roles for CD154/CD40 in the activation of skin-derived APCs and the development of Th1 cells in the skin-draining lymph nodes (sdLN). The presence of CD154 was important for optimal IL-12p40 and essential for Ag-specific IFN-, but CD154 expression by wild-type CD4– cells was insufficient to rescue recall responses of CD4+ cells from CD154–/– mice. This defect is probably due to impaired CD40-dependent IL-12 production in vivo, because administration of anti-CD40 Ab, or rIL-12, restored IFN- production by sdLN cells from CD154–/– mice. CD154 ligation of CD40 was not required for the migration of skin-derived APCs, but did have a limited role in their maturation (increased MHC II and CD86). Unexpectedly, although CD4 cells from CD154–/– mice were deficient in their ability to produce IFN-, they produced significant amounts of IL-4 and IL-5 in the presence of skin-derived APCs from wild-type and CD154–/– mice. Thus, in contrast to IFN-, the production of Th2-associated cytokines is (in this model) independent of CD154. We conclude that whereas the priming of Th1 responses soon after exposure to schistosome larvae is completely CD40/CD154 dependent, IL-4, IL-5, and IL-13 are independent of CD154, suggesting a dichotomy in the specific mechanisms that induce these cytokines by CD4+ cells in the sdLN

Isolation of bovine herpesvirus type 5 from the semen of a healthy bull in Australia.

Artificial insemination is widely used in the cattle industry and a major challenge is to ensure that semen is free of infectious agents. A healthy donor bull was tested for freedom from infectious agents. A bovine herpesvirus was isolated in testis cells and identified as bovine herpesvirus type 5 (BoHV-5) by polymerase chain reaction and by direct amplicon sequencing. The amplicon sequence shared 100% similarity with the published sequence of BoHV-5. This is the first report in Australia of BoHV-5 in semen. The implications of this finding are discussed

Detection and quantitation of gallid herpesvirus 1 in avian samples by 5′ taq nuclease assay utilizing minor groove binder technology

A 5′ Taq nuclease assay utilizing Minor Groove Binder technology and targeting the thymidine kinase gene of gallid herpesvirus 1 (GaHV-1) was designed and optimized for use in diagnosing avian infectious laryngotracheitis. The assay was specific for GaHV-1 in that it did not react with other avian viral or bacterial pathogens. The detection limit was 1.0-10-2 median tissue culture infectious dose per reaction or 90 target copies per reaction. Fifteen out of 41 diagnostic samples from sick birds reacted in the assay, five of which produced a typical alphaherpesvirus cytopathic effect (CPE) on chicken kidney (CK) cells. Sequencing, using amplicons generated by a polymerase chain reaction with primers flanking the 5′ Taq nuclease amplicon, confirmed the presence of GaHV-1 in six samples (two producing alphaherpesvirus CPE on CK cells, three not producing alphaherpesvirus CPE, and one that was not inoculated onto CK cells). Tracheal swabs taken from 18 healthy broilers did not react in the assay. The ability of the assay to determine viral load in samples was demonstrated. Overall the assay is suitable for the rapid diagnosis of infectious laryngotracheitis. © 2010 Houghton Trust Ltd

Pasteurella multocida detection by 5' Taq nuclease assay: A new tool for use in diagnosing fowl cholera

A 5′ Taq nuclease assay utilising minor groove binder technology and targeting the 16S rRNA gene was designed to detect Pasteurella multocida (the causative agent of fowl cholera) in swabs collected from poultry. The assay was first evaluated using pure cultures. The assay correctly identified four P. multocida taxonomic type strains, 18 P. multocida serovar reference strains and 40 Australian field isolates (17 from poultry, 11 from pigs and 12 from cattle). Representatives of nine other Pasteurella species, 26 other bacterial species (18 being members of the family Pasteurellaceae) and four poultry virus isolates did not react in the assay. The assay detected a minimum of approximately 10\ua0cfu of P. multocida per reaction. Of 79 poultry swabs submitted to the laboratory for routine bacteriological culture, 17 were positive in the 5′ Taq nuclease assay, but only 10 were positive by culture. The other 62 swabs were negative for P. multocida by both 5′ Taq nuclease assay and culture. The assay is suitable for use in diagnosing fowl cholera, is more rapid than bacteriological culture, and may also have application in diagnosing P. multocida infections in cattle and pigs

The integration of GPS, vegetation mapping and GIS in ecological and behavioural studies

Global Positioning System (GPS) satellite navigation receivers are increasingly being used in ecological and behavioural studies to track the movements of animals in relation to the environments in which they live and forage. Concurrent recording of the animal's foraging behaviour (e.g. from jaw movement recording) allows foraging locations to be determined. By combining the animal GPS movement and foraging data with habitat and vegetation maps using a Geographical Information System (GIS) it is possible to relate animal movement and foraging location to landscape and habitat features and vegetation types. This powerful approach is opening up new opportunities to study the spatial aspects of animal behaviour, especially foraging behaviour, with far greater precision and objectivity than before. Advances in GPS technology now mean that sub-metre precision systems can be used to track animals, extending the range of application of this technology from landscape and habitat scale to paddock and patch scale studies. As well as allowing ecological hypotheses to be empirically tested at the patch scale, the improvements in precision are also leading to the approach being increasing extended from large scale ecological studies to smaller (paddock) scale agricultural studies. The use of sub-metre systems brings both new scientific opportunities and new technological challenges. For example, fitting all of the animals in a group with sub-metre precision GPS receivers allows their relative inter-individual distances to be precisely calculated, and their relative orientations can be derived from data from a digital compass fitted to each receiver. These data, analyzed using GIS, could give new insights into the social behaviour of animals. However, the improvements in precision with which the animals are being tracked also needs equivalent improvements in the precision with which habitat and vegetation are mapped. This needs some degree of automation, as vegetation mapping at a fine spatial scale using the traditional manual approach is far too time consuming. This paper explores these issues, discussing new applications as well as approaches to overcoming some of the associated problems