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We investigated the spread of glycoprotein gE (gE)-negative pseudorabies virus (PRV) and its rescued 'wildtype' strain into and within the central nervous system (CNS) of 3-and 10-week-old pigs. This is the first study that demonstrates PRV invasion of the porcine CNS via the synapticaUy linked neurons of the olfactory and trigeminal routes and that demonstrates the role ofgE in this invasion. After intranasal inoculation with high doses of virus, gE-negative PRV replicated less efficiently in peripheral tissues. The titres of the gE-negative virus in the oropharyngeal mucosa, olfactory epithelium, draining lymph nodes and trigeminal ganglion were approximately 100-fold lower in 3-week-old pigs and 10-fold lower in 10-week-old pigs than titres of the ' wildtype' virus. In contrast to the 'wild-type' virus, titres of the gE-negative virus were very low or undetectable in the olfactory bulb, brain stem and other tissues of the CNS. Viral antigen of rescued 'wild-type' PRV and of gE-negative PRV was detected immunohistochemically in the olfactory epithelium and in neurons of the trigeminal ganglion, and also in the olfactory and trigeminal axons leading towards the CNS. But, in contrast to 'wild-type' virus, no viral antigen of the gEnegative virus was detected in second-or third-order neurons in the olfactory bulb or in the brain stem. We conclude that gE-negative PRV can infect first-order neurons of the olfactory and trigeminal routes and is able to spread via their axons towards the CNS. Yet, gEnegative PRV has a greatly reduced capacity to infect second-or third-order neurons. Finally, we report lateral spread of'wild-type' PRV in the trigeminal ganglion, i.e. nonsynaptic transport from neuron to neuron. Possible mechanisms that could explain the reduced levels of the gE-negative virus in the CNS are discussed.

Five malabsorption syndrome (MAS) homogenates from The Netherlands and Germany were used to reproduce MAS in broilers. We studied the histopathology after inoculation of 1-day-old broiler chicks and the agents that might be involved. Generally, the MAS homogenates induced signs that differed in severity and pathobiology. We could distinguish and classify the inoculated groups best by histopathology: proventriculitis, lesions in the small intestines in combination with proventriculitis, or lesions of the small intestines only. Lesions in the small intestine had more impact on weight gain depression than lesions in the proventriculus. In three out of five inoculated groups, microscopic lesions of the pancreas were found. Reovirus was detected in the inoculated groups by virus isolation and seroconversion, and reoviral antigen was detected by immunohistochemistry of the small intestine. Also, enteroviruslike particles were detected in three of the five inoculated groups, although not in the most affected group. Additionally, bacteriophages and bacteria (hemolytic Escherichia coli, Pasteurella hemolytica, and Enterococcus durans) were isolated from inoculated chicks. The role these agents play in pathogenesis of MAS is still unsolved.

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Envelope glycoprotein D (gD) is essential for entry of pseudorabies virus (PRV) into cells but is not required for the subsequent steps in virus replication. Phenotypically complemented gD mutants can infect cells and can spread, both in vitro and in mice, by direct cell-to-cell transmission. Progeny virions released by infected cellsare noninfectious because they lack gD. The aim of this study was to determine the role of gD in the neuropathogenicity of PRV in its natural host, the pig. We investigated whether gD-negative PRV can spread transneuronally via synaptically linked neurons of the olfactory and trigeminal routes. High doses of a phenotypically complemented gD mutant and gD mutants that are unable to express either gI or gI plus gE were inoculated intranasally in 3-to 5-week-old pigs. Compared with the wild-type virus, the virulence of the gD mutant was reduced. However, pigs inoculated with the gD mutant still developed fever and respiratory signs. Additional inactivation of either gI or gI plus gE further decreased virulence for pigs. Immunohistochemical examination of infected pigs showed that a PRV gD mutant could replicate and spread transneuronally into the central nervous system (CNS). Compared with the wild-type virus, the gD mutant had infected fewer neurons of the CNS on day 2. Nevertheless, on day 3, the gD-negative PRV had infected more neurons and viral antigens were present in second-and third-order neurons in the olfactory bulb, brain stem, and medulla oblongata. In contrast, gD mutants which are unable to express either gI or gI plus gE infected a limited number of first-order neurons in the olfactory epithelium and in the trigeminal ganglion and did not spread transneuronally or infect the CNS. Thus, transsynaptic spread of PRV in pigs can occur independently of gD. Possible mechanisms of transsynaptic transport of PRV are discussed.