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In alphaherpesviruses, glycoproteins E and I (gE and gI, respectively) form a heterodimer that facilitates cell-to-cell spread of virus. Using traditional homologous recombination techniques, as well as CRISPR/Cas9-assisted homologous recombination, we separately deleted gE and gI coding sequences from an Australian field strain (CSW-1) and a vaccine strain (A20) of infectious laryngotracheitis virus (ILTV) and replaced each coding sequence with sequence encoding green fluorescent protein (GFP). Virus mutants in which gE and gI gene sequences had been replaced with GFP were identified by fluorescence microscopy but were unable to be propagated separately from the wildtype virus in either primary chicken cells or the LMH continuous chicken cell line. These findings build on findings from a previous study of CSW-1 ILTV in which a double deletion mutant of gE and gI could not be propagated separately from wildtype virus and produced an in vivo phenotype of single-infected cells with no cell-to-cell spread observed. Taken together these studies suggest that both the gE and gI genes have a significant role in cell-to-cell spread in both CSW-1 and A20 strains of ILTV. The CRISPR/Cas9-assisted deletion of genes from the ILTV genome described in this study adds this virus to a growing list of viruses to which this approach has been used to study viral gene function., (© 2022. The Author(s).)

Human herpesvirus 8 (HHV-8), the causative agent of Kaposi’s sarcoma, multicentric Castleman’s disease and primary effusion lymphoma, predominantly manifests in immunocompromised individuals. However, infection in immunocompetent individuals does occur. The prevalence of HHV-8 exposure in blood donors from non-endemic countries ranges between 1.2% and 7.3%. Nothing was known about the prevalence in Australian blood donors. Therefore, this study investigated the active and cumulative exposure of HHV-8 in this cohort. Plasma samples (n = 480) were collected from eastern Australian blood donors and were tested for HHV-8 DNA by qPCR, and for HHV-8 antibodies by two different ELISAs. Samples initially positive on either ELISA were retested in duplicate on both, and on a mock-coated ELISA. Any samples positive two or three out of the three times tested on at least one ELISA, and repeat negative on the mock-coated ELISA, were assigned as repeat positive. None of the 480 samples tested contained HHV-8 DNA. Serological testing revealed 28 samples (5.83%; 95% CI: 3.74−7.93%) had antibodies to HHV-8. There was no difference (p > 0.05) in seropositivity between sex or with increasing age. This is the first study to show serological evidence of cumulative HHV-8 exposure and no HHV-8 DNAemia within a select blood donor population in Australia. Our molecular and serological data is consistent with published results for blood donors residing in HHV-8 non-endemic countries, which shows the prevalence to be very low.

Common carp (Cyprinus carpio) are an invasive species of the rivers and waterways of south-eastern Australia, implicated in the serious decline of many native fish species. Over the past 50 years a variety of control options have been explored, all of which to date have proved either ineffective or cost prohibitive. Most recently the use of cyprinid herpesvirus 3 (CyHV-3) has been proposed as a biocontrol agent, but to assess the risks and benefits of this, as well as to develop a strategy for the release of the virus, a knowledge of the fundamental processes driving carp distribution and abundance is required. To this end, we developed a novel process-based modelling framework that integrates expert opinion with spatio-temporal datasets via the construction of a Bayesian Network. The resulting weekly networks thus enabled an estimate of the habitat suitability for carp across a range of hydrological habitats in south-eastern Australia, covering five diverse catchment areas encompassing in total a drainage area of 132,129 km 2 over a period of 17-27 years. This showed that while suitability for adult and subadult carp was medium-high across most habitats throughout the period, nevertheless the majority of habitats were poorly suited for the recruitment of larvae and young-of-year (YOY). Instead, high population abundance was confirmed to depend on a small number of recruitment hotspots which occur in years of favourable inundation. Quantification of the underlying ecological drivers of carp abundance thus makes possible detailed planning by focusing on critical weaknesses in the population biology of carp. More specifically, it permits the rational planning for population reduction using the biocontrol agent, CyHV-3, targeting areas where the total population density is above a "damage threshold" of approximately 100 kg/ha., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)