Detection and differentiation of bovine herpesvirus 1 and 5 using a multiplex real-time polymerase chain reaction.

A multiplex real-time PCR was developed for the detection and differentiation of two closely related bovine herpesviruses 1 (BoHV-1) and 5 (BoHV-5). The multiplex real-time PCR combines a duplex real-time PCR that targets the DNA polymerase gene of BoHV-1 and BoHV-5 and a real-time PCR targeting mitochondrial DNA, as a house-keeping gene, described previously by Cawthraw et al. (2009). The assay correctly identified 22 BoHV-1 and six BoHV-5 isolates from the Biosecurity Sciences Laboratory virus collection. BoHV-1 and BoHV-5 were also correctly identified when incorporated in spiked semen and brain tissue samples. The detection limits of the duplex assay were 10 copies of BoHV-1 and 45 copies of BoHV-5. The multiplex real-time PCR had reaction efficiencies of 1.04 for BoHV-1 and 1.08 for BoHV-5. Standard curves relating Ct value to template copy number had correlation coefficients of 0.989 for BoHV-1 and 0.978 for BoHV-5. The assay specificity was demonstrated by testing bacterial and viral DNA from pathogens commonly isolated from bovine respiratory and reproductive tracts. The validated multiplex real-time PCR was used to detect and differentiate BoHV-1 and BoHV-5 in bovine clinical samples with known histories.

Biodiversity of Australian smut fungi.

There are about 250 species of smut fungi known from Australia of which 95 are endemic. Fourteen of these endemic species were first collected in the period culminating with the publication of Daniel McAlpine's revision of Australian smut fungi in 1910. Of the 68 species treated by McAlpine, 10 were considered to be endemic to Australia at that time. Only 23 of the species treated by McAlpine have names that are currently accepted . During the following eighty years until 1990, a further 31 endemic species were collected and just 11 of these were named and described in that period. Since 1990, 50 further species of endemic smut fungi have been collected and named in Australia . There are 115 species that are restricted to either Australia or to Australia and the neighbouring countries of Indonesia, New Zealand, Papua New Guinea and the Philippines . These 115 endemic species occur in 24 genera, namely Anthracoidea (1 species), Bauerago (1), Cintractia (3), Dermatosorus (1), Entyloma (3), Farysporium (1), Fulvisporium (1), Heterotolyposporium (1), Lundquistia (1), Macalpinomyces (4), Microbotryum (2), Moreaua (20), Pseudotracya (1), Restiosporium (5), Sporisorium (26), Thecaphora (2), Tilletia (12), Tolyposporella (1), Tranzscheliella (1), Urocystis (2), Ustanciosporium (1), Ustilago (22), Websdanea (1) and Yelsemia (2). About a half of these local and regional endemic species occur on grasses and a quarter on sedges . The northern tropical savannah region of Australia offers most promise for the discovery of new endemic species . The agricultural, quarantine and environmental significance to Australia of some introduced species is discussed.