Understanding the ecology and epidemiology of tick-borne diseases requires detailed knowledge of the complex interactions among the tick vector, the microorganisms they carry and the vertebrate hosts used by ticks, as well as the environmental conditions experienced by all three groups of organisms in this triad. In this thesis, I addressed questions relating to the biology and vector ecology of the Rocky Mountain wood tick (Dermacentor andersoni) and the American dog tick (Dermacentor variabilis). Comparisons were made of the distribution of both tick species, the vertebrate hosts used by immature ticks, and the types and prevalence of bacteria in individual ticks from multiple localities near the northern extent of their geographic ranges in western Canada. The results revealed that the distributions of both D. andersoni and D. variabilis have expanded since the 1960s, and there is now a broad zone of sympatry in southern Saskatchewan. In this zone of sympatry, D. andersoni and D. variabilis immatures were found to use the same species of small mammals as hosts and, in some cases, the same host individuals. This provides for the possibility of cross-transmission of bacteria from one tick species to the other. Bacteria of several genera (e.g. Rickettsia, Francisella, Arsenophonus and Anaplasma) were detected in D. andersoni and/or D. variabilis, some of which represented new tick-bacteria associations. However, most bacterial species were highly host (tick)-specific, except for three examples of apparent host switching from one tick species to the other at localities where the two tick species occurred in sympatry. The findings of this thesis provide a basis for understanding microbial transmission, the structure of tick-borne microbial communities, the risk of tick-borne disease in humans and animals, and the vector potential of D. andersoni and D. variabilis in geographical areas where they have not been studied previously.

Feral swine (Sus scrofa domesticus) are spreading across North America at an alarming rate. Four Canadian provinces and 39 states within the continental United States now report standing populations of feral pigs. Estimates place the number of feral hogs in Texas >2M, accounting for more than half of the United States population. It is known that feral swine impact local ecology following establishment, with regard to shifts in local flora and fauna. The overall objective of this research was to investigate the role of feral swine in tick-borne pathogen transmission in Texas. The underpinning objectives were to establish host records for tick species parasitizing feral swine, determine the species assemblages associated with feral swine among different ecoregions of Texas, determine by immunoassay to which tick-borne bacteria feral pigs were being exposed, and detect the DNA of tick-borne bacteria by polymerase chain reaction assay in the event of poor or early immune response by the host. Feral pigs (N=432) were harvested from June 2008 to June 2010 using box and corral traps and by aerial gunning. Seven species of ticks, Amblyomma americanum, A. cajennense, A. maculatum, Dermacentor albipictus, D. halli, and D. variabilis; and Ixodes scapularis, were collected. Immature stages of A. cajennense and A. americanum were collected as well. All classes of feral pigs, gender by age, were infested with ticks. Serum was collected through a multi-organizational effort from 2006 to 2010 and tested by ELISA for previous exposure to tick-borne pathogens in the genera Rickettsia and Ehrlichia (N=888) and Borrelia (N=849). Prevalence percentages by immunoassay were 27.59 percent, 13.18 percent and 2.12 percent for Rickettsia, Ehrlichia, and Borrelia, respectively. Samples positive by ELISA for exposure to Borrelia were further screened by Western Blot for exposure to Borrelia turicatae. The results were equivocal in most cases. Blood samples (N=233) were collected from 2008 to 2010 and analyzed by polymerase chain reaction for the detection of the DNA of these same three genera of bacteria. Two of the samples were positive by PCR for the presence of Borrelia DNA. Texas feral swine are serving as hosts for at least seven species of ticks and are interacting with tick-borne pathogen transmissions cycles in Texas.

Thesis (MSc (Veterinary Tropical Diseases)--University of Pretoria, 2009. / Includes bibliographical references. Also available in print format.

In 1994 and 1995, seven cases of Human monocytic ehrlichiosis were reported in Indiana; six cf these were from southern counties. To test whether Ehrlichia cha ffeensis is present in native ticks, adult ticks representing two species, Dermacentor variabilis and Amblyomma americanum were collected in southern Indiana during the first week of May, 1995 and tested by PCR. A total of 510 D. variabilis ticks were collected and placed into 102 pools (5 ticks per pool). A total of 430 adult A. americanum ticks were collected and placed into 88 pools of 5 ticks or less.D. variabilis ticks were used to optimize the isolation of PCR amplifiable DNA and determine the minimum number of bacterial cells detected. A modified CTAB-phenol method permitted the detection of as few as 100 bacterial cells. The optimal amount of isolated DNA for a PCR amplification was determined to be 2.7 pg of total nucleic acid.The 88 pools of adult A. americanum were subjected to DNA isolation, PCR amplification, and Southern analysis to determine the presence of E. chaffeensis bacteria. Using the 16S rRNA gene from E. chaffeensis, with species specific primers Hut and HE3. Of the 88 pools (430 ticks; 21 were determined to be positive for the presence E. chaffeensis bacteria. Thus, minimal field infection rate for adult A. americanum ticks in Southern Indiana was 4.88%. This calculation is based upon the assumption that at least one tick was positive in each positive pool. / Department of Biology

The genomic RNA encoding the structural proteins of louping ill, a tickborne flavivirus, was cloned and sequenced. Sequence comparisons of louping ill envelope protein showed greater homology with tick-borne than mosquito-borne flaviviruses and greater homology with the western than the far eastern subtype of tick-borne encephalitis virus. Louping ill and tick-borne encephalitis viruses are probably varieties of a common tick-borne ancestral virus. The average amino acid sequence diversity between members of the tick-borne serogroup was significantly lower than that of mosquito-borne serogroups, suggesting that tick-borne flaviviruses have been subjected to different evolutionary immune selection pressure from the mosquito-borne viruses. Using the published model of tick-borne encephalitis envelope protein and the derived sequence data on louping ill virus, three discontinuous peptides (amino acids 81-88, 207-212 and 230-234) which may represent critical molecular determinants within the receptor binding site of tick-borne flaviviruses, were identified. These peptides may provide a specific genetic marker for these viruses. Recombinant baculoviruses and vaccinia viruses containing cloned DNA, encoding either the envelope protein or the structural proteins of louping ill virus, were constructed. Glycosylated envelope protein, presented both inside and on the surface of insect and mammalian cells, was expressed by all four recombinant viruses. Differences in antigenic presentation of envelope protein were observed between envelope protein and structural protein constructs as well as between insect cell and mammalian cell expression systems. Despite the expression of epitopes known to elicit neutralizing and protective antibodies when present in authentic antigen, the recombinant envelope protein expressed by either baculovirus or vaccinia virus failed to induce, under the experimental conditions employed, either neutralizing or protective antibodies in both mice and rabbits against louping ill virus. Hence, louping ill envelope protein expressed by baculoviruses and vaccinia viruses was antigenically reactive but immunogenically inert.

Thesis (M.S.)--University of Missouri--Columbia, 2004. / "May, 2004." Typescript. Includes bibliographical references (leaves 35-38). Also issued on the Internet.

Thesis (MSc (Veterinary Tropical Diseases, Veterinary Science))--University of Pretoria, 2008. / Includes bibliographical references. Also available in print format.

Thesis (M.S.)--University of Missouri--Columbia, 2004. / Typescript. Includes bibliographical references (leaves 35-38). Also issued on the Internet.

The proposed thesis contributes to the knowledge about tick-borne encephalitis and its pathogenesis. The thesis describes pathogenesis and immunopathogenesis of tick-borne encephalitis, impact of host's genotype in clinical course determination, immune response of patients with acute tick-borne encephalitis, the mechanism of tick-borne encephalitis virus migration into central nervous system and virus interaction with cells of neurovascular unit as well as potential medical interventions.

This study was focused on the influence of hard tick from Ixodes ricinus on the replication of tick-borne encephalitis virus in vivo and the on the accompained changes in the levels of inflammatory cytokines which were determined in serum of infected mice. Futher the effect of tick cystatins, the inhibitors of cysteine proteases, on gene expression of interferon regulated factors was investigated in dendritic cells upon stimulation with Toll-like receptor ligands.