THE GLYCOPROTEINS OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS AND THEIR ROLE IN INFECTION AND IMMUNITY

The porcine reproductive and respiratory syndrome virus (PRRSV) is an economically important pathogen of swine and is known to cause abortion and infertility in pregnant sows and respiratory distress in piglets. PRRSV contains a major glycoprotein (GP5) and three minor glycoproteins (GP2a, GP3, and GP4) on the virion envelope, all of which are required for infectious virus production. To study their interactions amongst each other and with a cellular receptor for PRRSV, CD163, I cloned each of the viral glycoproteins and CD163 in various expression vectors. My studies have shown that while the GP2a, GP3, and GP4 are co-translationally glycosylated, the GP5 is post-translationally glycosylated. By using co-immunoprecipitation (co-IP) assays, strong interaction was demonstrated between GP4 and GP5 proteins, although weak interactions among the other envelope glycoproteins were also detected. Further, GP4 was found to mediate interactions leading to formation of multiprotein glycoprotein complex. My results also show that GP2a and GP4 proteins are the only two GPs that specifically interact with the CD163 molecule and that glycosylation of these GPs is required for efficient interaction. Based on these studies, I have developed an interactome map of the viral GPs and CD163 and have proposed a model of the viral glycoprotein complex and its interaction with CD163. Studies reported here also show that glycan addition at residue 184 (N184) of GP2a, and residues N42, N50, and N131 of GP3 is essential for recovery of infectious virus. Although single site glycosylation mutants of GP4 had no effect on infectious virus production, introduction of double mutations was lethal. The loss of glycan moieties of GP2a, GP3, and GP4 proteins had no effect on host neutralizing antibody production. Overall, I conclude that the PRRSV glycoproteins are co-translationally and post-translationally glycosylated, the GP4 protein is central to mediating interglycoprotein interactions, and along with GP2a, serves as the viral attachment protein that is responsible for interactions with the viral receptor, CD163. Further, glycosylation of GP2a, GP3, and GP4 proteins is required for infectious virus production, efficient interaction with CD163, but does not play any role in neutralizing antibody response in infected animals.

SEASONAL, HABITAT AND SEX-SPECIFIC PATTERNS OF FOOD UTILIZATION BY RED-WINGED BLACKBIRDS (Agelaius phoeniceous) IN EASTERN ONTARIO AND THEIR ECONOMIC IMPORTANCE

Crop depredation by red-winged blackbirds (Agelaius phoeniceus) causes serious economic losses to agricultural crops each year in both Canada and the United States. The concentration of vulnerable, monocultural crops, particularly corn, during periods when large flocks of blackbirds congregate in roosting areas prior to migration has invariably led to heavy feeding pressure (Stone et al., 1972; Wiens and Dyer, 1975; Tyler et al., 1978). Efforts to reduce damage levels by mechanical and chemical dispersal agents have been largely unsuccessful, at least in terms of a long-term solution to the problem. Recently, the lethal control of blackbird populations using surfactants has been proposed. However, the potential repercussions of the removal of substantial numbers of birds from northern breeding areas are virtually unknown (Robertson et al., 1978). Much of the research dealing with the feeding ecology of red-winged blackbirds has been limited to fall and winter periods when large aggregations of birds are actively involved in crop depredation (Goddad, 1969; Williams, 1976; Dolbeer et al., 1978) or pose a potential health hazard (Monroe and Cronholm, 1976). However, what is not known is the degree to which the removal of deleterious weed seed and insect pests cited in several studies (Bird and Smith, 1964; Mott et al., 1972; Robertson et al., 1978) might be of potential value to agriculture. The issue of whether the benefits derived from redwing foraging compensate for the negative aspects associated with crop depredation and health hazards remains largely unresolved. The present study attempted to evaluate the pest status of this species using diet information derived from food habits analysis conducted during the residency of red- winged blackbirds in a northern breeding area. By determining how the feeding ecology of red-winged blackbirds varies on a seasonal basis, among different breeding habitats and between sexes, we hoped to determine more realistically which segments of the population might be responsible for the greatest benefits or detriments and, thereby, more accurately evaluate the economic impact of the species as a whole. To achieve this aim, the study provides an accurate description of the common insects and weed pests utilized by redwings. By determining the relative proportions of those items known to be detrimental, we hoped to illustrate, at least qualitatively, the degree to which redwing foraging is comprised of both beneficial and harmful components.