Thesis (Ph. D.)--Virginia Polytechnic Institute and State University, 1990. / Vita. Abstract. Includes bibliographical references (leaves 111-123). Also available via the Internet.

Thesis (M.S.)--University of Wisconsin--Madison, 1962. / Typescript. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 45-54).

Thesis (M.S.)--University of Wisconsin--Madison, 1970. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

No description available.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1949. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves [i]-viii).

Brucellosis is endemic in many livestock worldwide especially developing countries. The aims of this study were to estimate the seroprevalence of bovine brucellosis and risk factors associated with the seropositivity in rural and peri-urban buffaloes and cattle populations of Sindh. Firstly, a cross sectional study was conducted to estimate the seroprevalence of bovine brucellosis in cattle and buffaloes of Sindh province, Pakistan. Serum samples (2600) were tested using Rose Bengal Plate Test. The overall seroprevalence of brucellosis in Sindh province was 13.96% (95% C.I.; 11.55 - 16.37). Of the 917 herds tested, 232 or 25.30% herds (95%C.I.; 22.51-28.24) were positive for brucellosis. The adult animals were 2.05 (95% C.I.; 1.14-3.68, P= 0.02) times more likely to test positive for brucellosis. The animals in a peri-urban dairy production system were 2.07 times (95%C.I.; 1.09-3.90, P = 0.03) times more likely have brucellosis. The species or sex of animal did not appear to affect the risk of seropositivity in cattle or buffalo in this population. Secondly, a cross sectional survey was conducted to understand the structure and composition of farms, animal husbandry and management practices in peri-urban dairy colonies in Karachi and farmers’ awareness of zoonoses. The mean herd size was 93.58 animals and 88.01% of these animals were female buffaloes. Of 326 farms surveyed, only 37.42% were able to associate animals with transmission of diseases in human. The characteristics of peri-urban dairy farms in Karachi are discussed. Thirdly, the value of FTA® cards in detecting the Brucella DNA in milk samples was estimated by determining the detection limits of genus specific ERI PCR assay for FTA® cards and comparing the PCR results from whole sediments taken from culturing pooled milk samples with taking sediment on FTA® cards. The detection limits of this method ranged from 6.6 x 103 cfu/ml for B. abortus to 7.17 x 106 cfu/ml for B. suis. Assuming the results of ERI PCR for the whole sediment as gold standard (method 1), the method using sediment on FTA® cards as test samples (method 2) showed a diagnostic sensitivity of 81.44% (95% C.I.; 75.54-87.33) but a poor diagnostic specificity of 42.86% (95%C.I.; 16.95-68.78). The kappa value, κ, was 0.14 (p = 0.02) demonstrating a poor agreement between the two methods. Lastly, 181 bulk milk samples were used to estimate the herd level prevalence of bovine brucellosis in Landhi dairy colony, Karachi. The ERI PCR was used to test these samples. The herd prevalence was estimated as 92.26% (95% C.I.; 88.34-96.19). For each level (50 animals) increase in herd size, the risk of herd being brucellosis positive increases by 2.38 times. The herds that have a male animal for breeding are 0.09 times less likely to have brucellosis. The history of abortion, presence of small ruminants or the regions of animal purchase don’t appear to have any association with the risk of brucellosis at a herd level in this population at LDC, Karachi. A high seroprevalence of bovine brucellosis in livestock population in Sindh and a very high herd level prevalence in peri-urban dairy farms in particular poses a serious threat to the public health and livestock production in Sindh, Pakistan.

Previous analysis of the bovine SLC11A1 complementary DNA (cDNA) failed to identify any nucleotide variations other than a microsatellite length variation within the 3' untranslated region functionally associated with bovine brucellosis. In this study I set out to identify mutations in the genomic complement of the gene that may be associated with resistance or susceptibility to bovine brucellosis, and to determine if the microsatellite length polymorphism in the 3'UTR of bovine SLC11A1 modulates gene expression and subsequent disease resistance in a phase dependent manner. The results of this study demonstrate that there are seventy-five total single nucleotide polymorphic (SNP) sites (excluding indels) located within the bovine genomic SLC11A1 sequence of a Brucella abortus resistant bull and a susceptible cow. Twenty of these SNPs segregated between resistant and susceptible populations, with 3 non-synonymous SNPs significantly associating with resistance or susceptibility to B. abortus infection. An A695G within exon 2 resulted in a histidine (resistant allele) to arginine (susceptible allele) amino acid substitution and was in significant linkage disequilibrium with the previously described 3' untranslated region (UTR) microsatellite length variation associated with brucellosis resistance. A transcriptional element search in the 3' UTR revealed a ETS-domain PU.1 site, an IFN-γ activation site (GAS), an Interferon Consensus Sequence Binding Protein site (ICSBP) and several Initiation Response sites (Inr), suggesting a possible function for this region in regulation of the expression of SLC11A1. A mobility shift assay confirmed sequence-specific DNA-protein interaction within this region. A luciferase reporter assay indicated that the 3'UTR of SLC11A1 could act as a downstream enhancer for expression. Macrophage killing assays with RAW264.7 cells expressing bovine SLC11A1 demonstrated that the microsatellite repeat is functionally associated with the macrophage killing efficiency, but not in a phase-dependent manner, suggesting that these length polymorphisms do not affect the angular orientation between cooperatively binding transcription factors, and leaves the possibility that the 3'UTR microsatellites regulate SLC11A1 transcription through some alternate mechanism, possibly mRNA stability.

No description available.

Brucellosis is a zoonotic disease caused by a Gram-negative coccobacillus. There are four Brucella strains of zoonotic importance in our domestic species, subdivided by their culture phenotypes: Brucella abortus (B. abortus, B. melitensis, B. suis (smooth strains) and B. canis (rough strain). Dogs can serve as hosts for all four of the zoonotic strains; however, routine serologic testing in dogs has been limited to the identification of antibodies to B. canis. The aim of our study was to identify a serologic test that can be utilized to identify smooth Brucella strain antibodies in canines. We hypothesize that the Brucella abortus Fluorescence Polarization Assay would be successful in identifying antibodies to smooth Brucella strain in canines. Ninety-five dogs, including forty-five hog hunting dogs were screened for circulating antibodies to any of the four zoonotic strains of the bacteria utilizing a combination of Canine Brucella Slide Agglutination Test (CBSA), Brucella canis Agar Gel Immunodiffusion II test (AGID), Brucella abortus Card Agglutination Test (BCA), and the Brucella abortus Fluorescence Polarization Assay (FPA). Test interpretation results yielded a 0% (0/95) smooth Brucella strain seropositivity rate, with 2% (2/95) of dogs yielding inconclusive rough Brucella strain serology results (0-2% rough strain seropositivity rate). Additionally, a retrospective portion of the study was performed to identify sera containing circulating antibodies to any of the smooth strains of Brucella by testing previously banked canine serum samples stored at Cornell's Veterinary Diagnostic Laboratory from 2018-2019 via Brucella abortus FPA. Of the 769 serum samples tested, 13/769 (1.7%) yielded an inconclusive result, 725/769 (94.2%) were negative, 30/769 (4%) yielded a positive FPA test result, and 1/769 (0.1%) had to be excluded due to insufficient sample remaining to perform the diagnostic test. Of the 30 FPA positive canine serum samples, 97% (29/30) also tested positive on the CBSA test. Additionally, there was a statistically significant (p <0.0001) likelihood of altered (spayed/neutered) and mixed breed dogs to be FPA positive when compared to intact, purebred dogs. / Master of Science / Brucellosis is a bacterial disease that can cause severe reproductive, orthopedic and general illness in both dogs and humans. There are four different species of Brucella that can be transmitted between animals and people: Brucella canis, abortus, suis, and melitensis. Although Brucella canis is the species that is widely recognized and breeding dogs are routinely tested for this strain, we have vastly under recognized the ability for dogs to contract and transmit the other three (smooth) Brucella species. Of added concern is the fact that the test currently used to screen dogs for brucellosis only identifies Brucella canis infection. Thus, veterinarians may be missing cases where dogs are infected with other Brucella species. This study revealed promising evidence in identification of smooth Brucella strain antibodies in canines, particularly altered and mixed breed canines, via the Brucella abortus Fluorescent Polarization Assay. The contributions of this study are threefold. First, to heighten awareness that both smooth and rough strains of brucellosis infection in dogs are infectious diseases of zoonotic concern. Second, it demonstrates that smooth Brucella strain infection along with the traditional strategy of selectively screening dogs breeding dogs may be underestimating the prevalence of brucellosis among the canine population in the United States thus, supporting the need for broadened screening recommendations. Third, it reveals the need for a commercially-available, validated test for the smooth strains of brucellosis in dogs and offers direction for future research efforts to likely focus on the validation of the Brucella abortus Fluorescent Polarization Assay.

B. abortus is an intracellular facultative bacterial pathogen which causes abortion in cattle and undulant fever in humans. Cattle vaccines such as B. abortus strains 19 and RB51 are live vaccine strains which protect approximately 75% of the vaccinated animals. No effective vaccines are available for the prevention of brucellosis in humans. We are developing vaccinia virus recombinants expressing various B. abortus proteins to prevent brucellosis in susceptible mammalian species. In this work the B. abortus groEL gene encoding the antigenic heat shock protein GroEL was subcloned into vaccinia virus via homologous recombination. Expression of the GroEL protein in vaccinia infected cells in-vivo was confirmed by immunoblotting. Groups of 5 female BALB/C mice were injected with the vaccinia recombinant or appropriate positive and negative control vaccines. Mice were bled and their humoral immune responses assessed. In addition, mice were challenged with virulent B. abortus strain 2308 and protection measured by the rate of splenic clearance of live Brucella. In spite of demonstrating specific GroEL antibodies in recombinant vaccinia injected mice, no significant level of protection was demonstrable. Preliminary lymphocyte transformation assays were carried out to establish if a cell mediated immune response to GroEL was induced in the vaccinated animals. / Master of Science