Microbial adaptation and change: avian influenza.

The evolution of influenza is a continuing process involving viral and host factors. The increasing frequency of emergence of the highly pathogenic H5N1, H7N3 and H7N7 influenza viruses and the panzootic spread of H9N2 influenza virus, all of which can be potentially transmitted to humans, are of great concern to both veterinary and human public health officials. The question is how soon the next pandemic will emerge. A convergence of factors, including the population densities of poultry, pigs and humans, are likely factors affecting the evolution of the virus. Highly concentrated poultry and pig farming, in conjunction with traditional live animal or 'wet' markets, provide optimal conditions for increased mutation, reassortment and recombination of influenza viruses. Strategies to reduce the evolution of influenza and the emergence of pandemics include the separation of species, increased biosecurity, the development of new vaccine strategies and better basic knowledge of the virus. More effective co-operation between scientists and veterinary and public health officials is required to achieve these goals.

Fate of plasmid DNA encoding infectious bursal disease virus VP2 capsid protein gene after injection into the pectoralis muscle of the chicken.

The objective of this study was to determine whether recombinant plasmid DNA injected intramuscularly into chickens expressed the gene of interest in vivo and could be subsequently detected in primary and secondary lymphoid tissues with polymerase chain reaction (PCR). The VP2 capsid protein gene of the standard challenge strain (STC) of infectious bursal disease virus (IBDV) was cloned into a eukaryotic plasmid, and purified DNA was prepared. Fourteen 2-wk-old chickens were injected in the pectoral musculature with 500 microg of plasmid DNA dissolved in sterile PBS. Seven chickens were similarly injected with PBS alone. Pectoral muscle, thymus, spleen, bursa of Fabricius, and cecal tonsils were collected at 12, 24, 36, 48, 72, 96, and 168 h postinjection for detection of protein expression (in muscle) and to extract total DNA for PCR amplification of the VP2 capsid gene. Expression of VP2 was demonstrated in muscle tissue at 12 and 24 h postinjection by using an indirect immunofluorescence assay. PCR amplification with primers specific for the VP2 gene showed that the DNA was present in the thymus, spleen, and bursa of Fabricius but not in cecal tonsils. These results demonstrate that plasmid DNA injected directly into the pectoral muscle of chickens is transcribed and translated at the injection site and promptly distributed to primary and secondary lymphoid tissues.