Thesis (Ph. D.)--University of Washington, 2008. / Vita. Includes bibliographical references (p. 144-163).

Chronic wasting disease (CWD) was detected in Saskatchewan wild deer populations in 2000 which prompted disease management actions consisting of population reduction. Little is known about population structure, health status, interactions or movement patterns of deer in Saskatchewan and these factors are important in designing a management program for CWD. As part of an ongoing study on deer movement patterns of wild deer in southern Saskatchewan, a survey was conducted to: 1) determine prevalence of CWD and selected infectious agents in mule deer (Odocoileus hemionus) and white-tailed deer (Odocoileus virginianus), and 2) identify infectious agents which could be used as a surrogate measure of the effectiveness of the adopted CWD management strategies. Tonsil biopsies, feces and blood were collected from 254 mule deer and 43 white-tailed deer during winters of 2006, 2007 and 2008. Immunohistochemical staining of tonsil biopsies for CWD revealed a prevalence of 2.4% (6/249) in mule deer and 0% (0/43) in white-tailed deer. Parasitological investigation of 253 fecal samples from mule deer identified eggs of nematodes in the superfamily Trichostrongyloidea (29.2%); and parasitic stages of the following genera: Nematodirus (7.1%), Skrjabinema (14.3%), Trichuris (0.8%), Moniezia (16.2%), Thysanosoma (12.2%), Orthostrongylus (35.2%), Eimeria (13.4%) and Giardia (0%, 0/137). A similar investigation of 42 white-tailed deer fecal samples identified parasitic stages of nematodes in the super family Trichostrongyloidea (4.8%) and in genera of Orthostrongylus (2.4%), Moniezia (2.4%) and Eimeria (2.4%). Dorsal-spined larvae were detected in 2.4% of the white-tailed deer fecal samples. In serum samples from 253 mule deer, antibodies (Ab) were detected against bovine herpesvirus1 (BoHV-1) (34.8%), parainfluenza-3 (PI-3) (56.5%), bovine virus diarrhoea virus (BVDV-1) (30.8%) and Neospora caninum (15.4%, 36/245). In serum samples from 40 white-tailed deer, Ab to BoHV-1(32.5%), PI-3 (35%), BVD-1 (12.5%) and Neospora caninum (20.5%, 8/39) was detected. Based on relative host specificity, moderate prevalence and horizontal routes of transmission, herpesvirus, parainfluenza 3, Eimeria and Skrjabinema were identified as infectious agents which could potentially be used to evaluate the effectiveness of disease management strategies, which may in turn predict the response of CWD to these same strategies. Using polymerase chain reaction (PCR) a herpesvirus was detected, in 42.1% (40/95) of retropharyngeal lymph nodes from hunter-submitted mule deer and white-tailed deer heads from Saskatchewan in 2007. DNA sequences of the partial DNA polymerase gene from this virus were 98 - 100% identical to mule deer lymphotropic herpesvirus (mule deer-LHV). A 3.6 kb contiguous sequence of mule deer-LHV genome was generated by genome walking (GenBank Accession number: HM014314). Use of a mule deer-LHV-specific PCR on buffy coat samples collected during winters of 2007 and 2008, detected mule deer-LHV in 42.1% (67/158) of mule deer and 33.3% (8/24) of white-tailed deer. Very little DNA sequence diversity in the partial sequences of glycoprotein B (gB) gene and the intergenic spacer regions between DPOL and gB gene of mule deer-LHV was observed among deer from different wildlife management zones. Mule deer-LHV is also a potential marker for evaluating the effectiveness of disease management activities because of its moderate prevalence, host specificity, ease of sample collection and the availability of a rapid and low-cost method for its detection. A variable region of the mule deer-LHV genome needs to be identified if this virus to be used as an inferential tool for studying host population structure.

Chronic wasting disease (CWD) was detected in Saskatchewan wild deer populations in 2000 which prompted disease management actions consisting of population reduction. Little is known about population structure, health status, interactions or movement patterns of deer in Saskatchewan and these factors are important in designing a management program for CWD. As part of an ongoing study on deer movement patterns of wild deer in southern Saskatchewan, a survey was conducted to: 1) determine prevalence of CWD and selected infectious agents in mule deer (Odocoileus hemionus) and white-tailed deer (Odocoileus virginianus), and 2) identify infectious agents which could be used as a surrogate measure of the effectiveness of the adopted CWD management strategies. Tonsil biopsies, feces and blood were collected from 254 mule deer and 43 white-tailed deer during winters of 2006, 2007 and 2008. Immunohistochemical staining of tonsil biopsies for CWD revealed a prevalence of 2.4% (6/249) in mule deer and 0% (0/43) in white-tailed deer. Parasitological investigation of 253 fecal samples from mule deer identified eggs of nematodes in the superfamily Trichostrongyloidea (29.2%); and parasitic stages of the following genera: Nematodirus (7.1%), Skrjabinema (14.3%), Trichuris (0.8%), Moniezia (16.2%), Thysanosoma (12.2%), Orthostrongylus (35.2%), Eimeria (13.4%) and Giardia (0%, 0/137). A similar investigation of 42 white-tailed deer fecal samples identified parasitic stages of nematodes in the super family Trichostrongyloidea (4.8%) and in genera of Orthostrongylus (2.4%), Moniezia (2.4%) and Eimeria (2.4%). Dorsal-spined larvae were detected in 2.4% of the white-tailed deer fecal samples. In serum samples from 253 mule deer, antibodies (Ab) were detected against bovine herpesvirus1 (BoHV-1) (34.8%), parainfluenza-3 (PI-3) (56.5%), bovine virus diarrhoea virus (BVDV-1) (30.8%) and Neospora caninum (15.4%, 36/245). In serum samples from 40 white-tailed deer, Ab to BoHV-1(32.5%), PI-3 (35%), BVD-1 (12.5%) and Neospora caninum (20.5%, 8/39) was detected. Based on relative host specificity, moderate prevalence and horizontal routes of transmission, herpesvirus, parainfluenza 3, Eimeria and Skrjabinema were identified as infectious agents which could potentially be used to evaluate the effectiveness of disease management strategies, which may in turn predict the response of CWD to these same strategies. Using polymerase chain reaction (PCR) a herpesvirus was detected, in 42.1% (40/95) of retropharyngeal lymph nodes from hunter-submitted mule deer and white-tailed deer heads from Saskatchewan in 2007. DNA sequences of the partial DNA polymerase gene from this virus were 98 - 100% identical to mule deer lymphotropic herpesvirus (mule deer-LHV). A 3.6 kb contiguous sequence of mule deer-LHV genome was generated by genome walking (GenBank Accession number: HM014314). Use of a mule deer-LHV-specific PCR on buffy coat samples collected during winters of 2007 and 2008, detected mule deer-LHV in 42.1% (67/158) of mule deer and 33.3% (8/24) of white-tailed deer. Very little DNA sequence diversity in the partial sequences of glycoprotein B (gB) gene and the intergenic spacer regions between DPOL and gB gene of mule deer-LHV was observed among deer from different wildlife management zones. Mule deer-LHV is also a potential marker for evaluating the effectiveness of disease management activities because of its moderate prevalence, host specificity, ease of sample collection and the availability of a rapid and low-cost method for its detection. A variable region of the mule deer-LHV genome needs to be identified if this virus to be used as an inferential tool for studying host population structure.

Herpesvirus ateles is an oncogenic tumor virus of New World primates, which has sequence homology and biological properties in common with Herpesvirus saimiri. Each causes acute T-cell lymphomas in susceptible species of New World primates, while establishing a latent infection of the T lymphocytes of its normal host. The thesis research consists of characterization of the viral genome, cloning of viral DNA, in vitro immortalization of T cells with the virus, and mapping of viral transcripts within immortalized cells. Additional experiments performed to transfect cloned DNA into immortalized cells were unsuccessful. Fragments of H. ateles virion DNA were cloned into the vector pHyg, which is selectable in both prokaryotic and mammalian cells. Overlapping clones of >95% of the viral genome were characterized by restriction mapping, and were used to determine restriction maps of H. ateles strains 73 and 810. Peripheral blood T lymphocytes from several species of New World primate were expanded in medium containing IL-2. T-cells from cottontopped tamarins were immortalized with H. ateles 73, H. saimiri 11, and H. saimiri 484-77, becoming IL-2-independent and growing continuously in culture. Immortalization was highly efficient, occurring reproducibly in cultures of 104-105 cells. Immortalization of IL-2-expanded T-cells of red-bellied tamarins, spider monkeys, and squirrel monkeys was unsuccessful with all strains of virus used. The right end of H. saimiri DNA is deleted in nononcogenic mutants. It has been found to produce four small RNAs in immortalized cells. Similarly, two small viral RNAs were found to be transcribed in cells immortalized by H. ateles 73. The RNAs, of 115 and 119 nuc1eotides, were mapped in the right end of the viral genome. The RNAs contain two regions of high conservation with H. saimiri RNAs. The genes for the small RNAs contain promoter, internal, and terminator sequences characteristic of cellular U RNAs.