Publications

Epizootic hemorrhagic disease virus (EHDV) is a Culicoides-transmitted orbivirus that infects domestic and wild ruminants and is provisionally thought to be distributed throughout Africa, North America, Australia, East Asia and the Middle East. Historically, of the seven proposed serotypes of EHDV, only EHDV-1 and EHDV-2 have been reported from North America. In 2006, EHDV isolates were recovered from moribund or dead white-tailed deer (Odocoileus virginianus) in Indiana and Illinois that could not be identified as either EHDV-1 or EHDV-2 by virus neutralization tests or by serotype-specific RT-PCR. Additional serological and genetic testing identified the isolates as EHDV-6, a serotype that, although originally described from Australia, has recently been recognized as an emerging pathogen of cattle in Morocco, Algeria and Turkey. In 2007 and 2008, EHDV-6 was isolated again from white-tailed deer, this time in Missouri, Kansas and Texas, suggesting that the virus is capable of overwintering and that it may become, or already is, endemic in a geographically widespread region of the USA. Genetic characterization of the virus indicates that it is a reassortant, such that the outer capsid proteins determining serotype specificity (VP2 and VP5) are derived from exotic EHDV-6, whilst the remaining structural and non-structural proteins are apparently obtained from indigenous EHDV-2 (Alberta).

Culicoides biting midges (Diptera: Ceratopogonidae) are responsible for the spread of several arboviruses of livestock and humans that are of international importance. This study assesses the virulence of 18 insect-pathogenic fungal strains from the genera Metarhizium, Beauveria, Isaria and Lecanicillium to larval stages of Culicoides nubeculous Meigen as a means of examining their potential as biocontrol agents. In initial screening, six strains of M. anisopliae (ERL700, CA1, V275, LRC181A, ARSEF 3291 and ARSEF 4556) outperformed the other tested genera and were found to cause between 90% and 100% larval mortality in all larval instars of this species at 72 h post inoculation. The virulence of the most effective strain, M. anisopliae V275, was then further tested by exposing larvae to doses which ranged from 104–108 conidia/ml and recording mortality at 24, 48 and 72 h in a 24-multi-well plate with each well containing 600 ?l of water and at 24 and 48 h in 250 ml plastic cups containing 50 ml of water. Sensitivity of larvae was extremely high in the multi-well plates, with LC50 values of 4.3–4.5 × 103 conidia/ml and no significant differences between larval instars. In the 250 ml cups, M. anisopliae V275 caused mortalities of between 70% and 100% to larvae and later instars exhibited higher mortality rates. The results are discussed in relation to incorporation of M. anisopliae into biocontrol programmes to control arboviruses vectored by Culicoides.

In common with other positive-strand RNA viruses, replication of feline calicivirus (FCV) results in rearrangement of intracellular membranes and production of numerous membrane-bound vesicular structures on which viral genome replication is thought to occur. In this study, bioinformatics approaches have identified three of the FCV non-structural proteins, namely p32, p39 and p30, as potential transmembrane proteins. These proteins were able to target enhanced cyan fluorescent protein to membrane fractions where they behaved as integral membrane proteins. Immunofluorescence microscopy of these proteins expressed in cells showed co-localization with endoplasmic reticulum (ER) markers. Further electron microscopy analysis of cells co-expressing FCV p39 or p30 with a horseradish peroxidase protein containing the KDEL ER retention motif demonstrated gross morphological changes to the ER. Similar reorganization patterns, especially for those produced by p30, were observed in naturally infected Crandel–Rees feline kidney cells. Together, the data demonstrate that the p32, p39 and p30 proteins of FCV locate to the ER and lead to reorganization of ER membranes. This suggests that they may play a role in the generation of FCV replication complexes and that the endoplasmic reticulum may represent the potential source of the membrane vesicles induced during FCV infection.

A network of foot and mouth (FMD) vaccine banks has been initiated with the support of vaccine bank managers and technical advisors that participated in a workshop held at the Institute for Animal Health, Pirbright, in the United Kingdom in April 2006. Terms of Reference that provide guidance for coordinated activities are under consultation. Practical and economic benefits can be realised from collaboration, which will be achieved through mutually acceptable mechanisms for the exchange of information and materials relevant to vaccine banks and their management. If administrative and technical hurdles can be overcome, the network has the potential to contribute significantly to the improved control of FMD worldwide. A 'global' and interactive vaccine bank association could be created by agreeing a system of resource sharing that could orchestrate additional emergency cover with vaccine or antigen from the reserves of network members.