Bacillus anthracis is a facultative intracellular bacterial pathogen that can cause cutaneous, gastrointestinal or respiratory disease in many vertebrates, including humans. Commercially available anthrax vaccines for immunization of humans are of limited duration and do not protect against the respiratory form of the disease. Brucella abortus is a facultative intracellular bacterium that causes chronic infection in animals and humans. As with other intracellular pathogens, cell mediated immune responses (CMI) are crucial in affording protection against brucellosis. B. abortus strain RB51 has been shown to be useful in eliciting protective cell mediated immunity and humoral responses against Brucella in cattle and other animal species. Since the protective antigen (PA) of B. anthracis is known to induce protective antibodies, it was decided that the objective of this research was to test whether the gene encoding PA could be expressed in Brucella producing a bivalent vaccine to protect against both brucellosis and anthrax. The pag gene was transcriptionally fused to promoters of genes encoding superoxide dismutase or heat shock protein groE, subcloned into a broad host range plasmid (pBBR1MCS) and shown to express in E. coli by immunoblotting using antiserum specific for PA. The immunoblot results revealed that E. coli produced a PA protein of the expected size. In addition, the culture medium was shown to contain the same PA protein using immunoblotting. These results show that E. coli is capable of expressing B. anthracis PA in both the cellular and extracellular forms. The pBB/PA plasmid was used to transform B. abortus RB51 and CmR clones screened for the expression of PA by immunoblotting. Twenty clones of strain RB51/pBBSOD were show to express a 30kDa PA protein. Three clones of strain RB51/pBBGroE-PA were shown to express a 63-83kDa protein as detected by antiserum specific for PA. Using the A/J mouse, an immunocompromised vertebrate model, immunization and challenge studies were performed. Preliminary results demonstrate that the bivalent vaccine is capable of producing protection against a live challenge with B. abortus and some protection against live non-disease producing spores of B. anthracis. / Master of Science

Call number: LD2668 .T4 1948 K4 / Master of Science

Call number: LD2668 .T4 1955 N67 / Master of Science

No description available.

In developing countries, cities are rapidly expanding, and urban and peri-urban agriculture (UPA) has an important role in feeding a growing urban population. However, UPA carries risks of zoonotic disease transmission. This study aims to understand the characteristics of UPA in Kampala, Uganda and the zoonotic risks to humans. Following a general overview of the subject in Chapter 1, Chapter 2 describes the determination of urban, peri-urban and rural areas of the Kampala economic zone and socio-economical characteristics of the peri-urban interface compared with the urban and rural counter parts using the Village Characteristic Survey in 87 randomly selected Local Councils (LC1s). Chapter 3 describes the characteristics of UPA in Kampala and found both the contribution of agriculture to the livelihood and risks of zoonoses were high. In Chapter 4, the most important zoonotic diseases affecting populations living in urban and peri-urban areas in Kampala were identified; brucellosis, GI infections, Mycobacterium bovis tuberculosis and Taenia solium cycticercosis based on investigations using the medical records of Mulago National Referral Hospital. Chapter 5 describes a series of case-control studies of the identified most important zoonoses using a spatial approach. The risks of identified zoonoses might be homogenously high at all levels of urbanicity. Brucellosis appeared to be the most significant disease. Chapter 6 investigates brucellosis further, with an epidemiological investigation into the prevalence of the disease in milking cows and a quantitative analysis of the level of infection in milk for sale in and around Kampala. The prevalence was 6.2% (95%CI: 2.7-9.8) at the herd level. Chapter 7 describes the risk analysis for purchase raw milk infected with Brucella abortus in urban areas of Kampala. A quantitative milk distribution model was developed synthesizing the results from the cattle survey and interviews with milk sellers. The infection rates of milk at sale obtained from milk testing and cattle survey were multiplied to this model to present distribution of the risk. 11.7% of total milk consumed in urban Kampala was infected when purchased and the risk management analysis found the most effective control option for human brucellosis was construction of milk boiling centres either in Mbarara, the largest dairy production area in Uganda, or in peri-urban areas of Kampala.

Tendo em vista a importância da bubalinocultura como fonte de proteína para o Brasil e também do Programa Nacional de Controle e Erradicação da Brucelose e da Tuberculose para a evolução das cadeias produtoras de bovinos e bubalinos e, devido à escassez de estudos sobre o sorodiagnóstico da brucelose na espécie bubalina, procedimento no qual se apóia o processo de certificação de rebanhos livres e monitorados, o presente estudo objetivou comparar o desempenho de diferentes testes para o sorodiagnóstico da brucelose nessa espécie. Soros de 696 fêmeas bubalinas adultas foram submetidos aos testes qualitativos BPA, AATE, AAT e quantitativos: SRP, SLT, ELISAI, ELISAC, PF e ME e FC. Foi empregada, gold standard, a combinação de dois testes, FC e ME, considerando dois critérios: a) animais positivos para os reagentes na FC (título &ge; 20UI) e no ME (critério para fêmea vacinada) e como negativos os animais não reagentes na FC (título < 20UI) e no ME; b) animais positivos os reagentes na FC (título &ge; =20UI) e no ME (critério para fêmea não vacinada) e negativos os não reagentes na FC (título < 20UI) e no ME. Esta consideração foi necessária em função dos animais não apresentarem histórico de vacinação confiável o que interfere na determinação do ponto de corte a ser utilizado no teste ME de animais vacinados e não vacinados. Desta forma, numa primeira análise todos os animais foram avaliados no ME como vacinados e em outra como não vacinados. Com base nesses resultados, foi construída a curva ROC para todos os testes quantitativos (SRP, SLT, ELISAC, ELISAI e PF). Com os resultados dessa análise estabeleceu-se pontos de corte que otimizam os valores de sensibilidade e especificidade relativas para cada teste. Esses valores foram utilizados para o cálculo do índice de concordância Kappa para cada teste, incluindo os qualitativos. Concluiu-se que: a) os resultados dos estudos com os testes de ELISAC e PF em bovinos podem ser inferidos para bubalinos com razoável segurança; b) devem ser estabelecidos pontos de corte específicos para búfalos para o SLT, o SRP e o ELISAI; c) as melhores combinações de sensibilidade e especificidade relativas foram alcançadas pelo ELISAC, o PF, o BPA e o AAT; d) os melhores resultados de Kappa foram verificados para o ELISAC (kappa), PF, BPA, AATE e AAT; e) o BPA, AAT e AATE são os melhores testes para o sorodiagnóstico de triagem em bubalinos e o ELISAC e o PF são os mais promissores testes confirmatórios nessa espécie, já que ganhos adicionais de especificidade podem ser alcançados pelo aumento do ponto de corte. / Considering the importance of the buffalo as a protein source in Brazil and the National Program of Brucelosis and Tuberculosis Control and Eradication for the evolution of the bovine and bubaline sector, and also due to the few number of studies on serological diagnosis of brucelosis in this specie, procedure that supports the process of certification of brucelosis free herds, the present study aimed to compare different serological tests for brucelosis diagnosis on buffaloes. Serum samples from 696 adult bubaline females had been analyzed to BPA, AATE, AAT (qualitatives), SRP, SLT, ELISAI, ELISAC, PF and ME and FC (quantitatives). The combination of two tests, FC and ME was used as reference, considering two criteria: a) positive animals: reactors for the FC (titer &ge; 20UI) and ME (vaccinated females), negative animals: no reactors for the FC (titer < 20UI) and ME (vaccinated females); b) positive animals: reactors for the FC (titer &ge; 20UI) and ME (not vaccinated females), negative animals: no reactors for the FC (titer < 20UI) and ME (not vaccinated females). It was necessary because was not possible to garantee the true vaccination status of the animals. The first step was performing the ROC analysis for all the quantitative tests (SRP, SLT, ELISAC, ELISAI and PF), and so the relative sensitivity and specificity and the Kappa were calculated for each test. Conclusions: a) the cut offs of ELISAC and PF tests proposed for bovines can be used for bubalines with reasonable security; b) specific cut offs for buffaloes to be used in SLT, SRP and ELISAI must be established; c) the best relative sensitivity and specificity had been reached by the ELISAC, PF, BPA and AAT; d) the best results of the Kappa had been reached by ELISAC (kappa), PF, BPA, AATE and AAT; e) BPA, AAT and AATE are the best screening diagnosis test for bubalines and ELISAC and PF are the most promising confirmatory tests for this species.

Não disponível / Not available

Tendo em vista a importância da bubalinocultura como fonte de proteína para o Brasil e também do Programa Nacional de Controle e Erradicação da Brucelose e da Tuberculose para a evolução das cadeias produtoras de bovinos e bubalinos e, devido à escassez de estudos sobre o sorodiagnóstico da brucelose na espécie bubalina, procedimento no qual se apóia o processo de certificação de rebanhos livres e monitorados, o presente estudo objetivou comparar o desempenho de diferentes testes para o sorodiagnóstico da brucelose nessa espécie. Soros de 696 fêmeas bubalinas adultas foram submetidos aos testes qualitativos BPA, AATE, AAT e quantitativos: SRP, SLT, ELISAI, ELISAC, PF e ME e FC. Foi empregada, gold standard, a combinação de dois testes, FC e ME, considerando dois critérios: a) animais positivos para os reagentes na FC (título &ge; 20UI) e no ME (critério para fêmea vacinada) e como negativos os animais não reagentes na FC (título < 20UI) e no ME; b) animais positivos os reagentes na FC (título &ge; =20UI) e no ME (critério para fêmea não vacinada) e negativos os não reagentes na FC (título < 20UI) e no ME. Esta consideração foi necessária em função dos animais não apresentarem histórico de vacinação confiável o que interfere na determinação do ponto de corte a ser utilizado no teste ME de animais vacinados e não vacinados. Desta forma, numa primeira análise todos os animais foram avaliados no ME como vacinados e em outra como não vacinados. Com base nesses resultados, foi construída a curva ROC para todos os testes quantitativos (SRP, SLT, ELISAC, ELISAI e PF). Com os resultados dessa análise estabeleceu-se pontos de corte que otimizam os valores de sensibilidade e especificidade relativas para cada teste. Esses valores foram utilizados para o cálculo do índice de concordância Kappa para cada teste, incluindo os qualitativos. Concluiu-se que: a) os resultados dos estudos com os testes de ELISAC e PF em bovinos podem ser inferidos para bubalinos com razoável segurança; b) devem ser estabelecidos pontos de corte específicos para búfalos para o SLT, o SRP e o ELISAI; c) as melhores combinações de sensibilidade e especificidade relativas foram alcançadas pelo ELISAC, o PF, o BPA e o AAT; d) os melhores resultados de Kappa foram verificados para o ELISAC (kappa), PF, BPA, AATE e AAT; e) o BPA, AAT e AATE são os melhores testes para o sorodiagnóstico de triagem em bubalinos e o ELISAC e o PF são os mais promissores testes confirmatórios nessa espécie, já que ganhos adicionais de especificidade podem ser alcançados pelo aumento do ponto de corte. / Considering the importance of the buffalo as a protein source in Brazil and the National Program of Brucelosis and Tuberculosis Control and Eradication for the evolution of the bovine and bubaline sector, and also due to the few number of studies on serological diagnosis of brucelosis in this specie, procedure that supports the process of certification of brucelosis free herds, the present study aimed to compare different serological tests for brucelosis diagnosis on buffaloes. Serum samples from 696 adult bubaline females had been analyzed to BPA, AATE, AAT (qualitatives), SRP, SLT, ELISAI, ELISAC, PF and ME and FC (quantitatives). The combination of two tests, FC and ME was used as reference, considering two criteria: a) positive animals: reactors for the FC (titer &ge; 20UI) and ME (vaccinated females), negative animals: no reactors for the FC (titer < 20UI) and ME (vaccinated females); b) positive animals: reactors for the FC (titer &ge; 20UI) and ME (not vaccinated females), negative animals: no reactors for the FC (titer < 20UI) and ME (not vaccinated females). It was necessary because was not possible to garantee the true vaccination status of the animals. The first step was performing the ROC analysis for all the quantitative tests (SRP, SLT, ELISAC, ELISAI and PF), and so the relative sensitivity and specificity and the Kappa were calculated for each test. Conclusions: a) the cut offs of ELISAC and PF tests proposed for bovines can be used for bubalines with reasonable security; b) specific cut offs for buffaloes to be used in SLT, SRP and ELISAI must be established; c) the best relative sensitivity and specificity had been reached by the ELISAC, PF, BPA and AAT; d) the best results of the Kappa had been reached by ELISAC (kappa), PF, BPA, AATE and AAT; e) BPA, AAT and AATE are the best screening diagnosis test for bubalines and ELISAC and PF are the most promising confirmatory tests for this species.

Thesis (M.S.)--University of Nevada, Reno, 2008. / "August, 2008." Includes bibliographical references (leaves 62-69). Online version available on the World Wide Web.

Thesis (MS)--Montana State University--Bozeman, 2009. / Typescript. Chairperson, Graduate Committee: Scott Creel. Includes bibliographical references (leaves 45-49).