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1
Smuts, Celia Margaretha. "Development of tools to improve the detection of Trypanoma evansi in Australia /." Murdoch University Digital Theses Program, 2009. http://wwwlib.murdoch.edu.au/adt/browse/view/adt-MU20090709.113425.
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Tchamo, Cesaltina da Conceicao Lopes Menete. "Evaluation of the pathogenicity in goats of Trypanosoma congolense from Matutuine, Mozambique." Pretoria : [s.n.], 2007. http://upetd.up.ac.za/thesis/available/etd-04212008-143822/.
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Knight-Jones, Theo. "Field evaluation of foot-and-mouth disease vaccination in Turkey." Thesis, Royal Veterinary College (University of London), 2014. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.618321.
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Yu, Chun-I. Palucka Karolina Banchereau Jacques. "Humanized mice to test vaccination against influenza virus via dendritic cells." Waco, Tex. : Baylor University, 2008. http://hdl.handle.net/2104/5184.
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Thesis (Ph.D.)--Baylor University, 2008.In abstract the '2' and '-/-' in NOD-SCID-[beta]2m-/- is superscript. In abstract the '+' after CD34 and CD8 is superscript. In abstract the '-' and '+' in CD45RA-CD27+CD4+ are superscript. Includes bibliographical references (p. 103-123).
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O'Quin, Jeanette M. "Prophylactic Animal Rabies Vaccination Requirements in Ohio and Involvement of Local Health Departments in Low Cost Rabies Vaccination Clinics." The Ohio State University, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=osu1322617160.
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La, Tom. "Characterisation, recombinant expression and immunogenicity of BHLP29.7, an outer membrane lipoprotein of Brachyspira hyodysenteriae." La, Tom (2006) Characterisation, recombinant expression and immunogenicity of BHLP29.7, an outer membrane lipoprotein of Brachyspira hyodysenteriae. PhD thesis, Murdoch University, 2006. http://researchrepository.murdoch.edu.au/140/.
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Swine dysentery (SD) is an important endemic infection in many piggeries, and control can be problematic. In this study, the gene encoding a 29.7 kDa outer membrane lipoprotein of the causative intestinal spirochaete Brachyspira hyodysenteriae, was identified and sequenced. An 816 bp hypothetical open reading frame (ORF) was identified, with a potential ribosome binding site, and putative -10 and -35 promoter regions upstream from the start of the ORF. The 29.7 kDa outer membrane lipoprotein was designated Bhlp29.7 and the encoding gene named bhlp29.7.
The amino acid sequence of Bhlp29.7 included a 19 residue hydrophobic signal peptide, incorporating a potential signal peptidase cleavage site and membrane lipoprotein lipid attachment site. In silico analysis of this protein together with lipidation studies further supported its probable outer membrane localisation. Comparison of the Bhlp29.7 sequence with public sequence databases showed that it had up to 40% similarity with the D-methionine substrate-binding outer membrane lipoprotein (MetQ) of a number of bacterial pathogens. The Bhlp29.7 gene was detected in all 48 strains of B. hyodysenteriae examined, and in Brachyspira innocens strain B256T, but not in 10 other strains of B. innocens or in 42 strains of other Brachyspira spp. The gene was sequenced from B. innocens strain B256T and from 11 strains of B. hyodysenteriae. The B. hyodysenteriae genes shared 97.9-100% nucleotide sequence identity and had 97.5-99.5% identity with the gene of B. innocens strain B256T. The Bhlp29.7 gene was subsequently cloned and expressed as a histidine fusion protein in an Escherichia coli expression system.
An ELISA test using recombinant his-tagged Bhlp29.7 (His6-Bhlp29.7) as the detecting antigen was developed and evaluated. The threshold value of the test was chosen to provide a highly stringent assessment of the disease status of a herd. The sensitivity and specificity of the test was 100%. When the test was applied to sera from eight herds with suspected SD, four gave ELISA values indicating that the herds were diseased. The remaining four herds gave ELISA values below the threshold value. These results indicated that the Bhlp29.7-ELISA was useful as an indirect test for exposure of a herd to B. hyodysenteriae and may be a helpful complement to current methods of SD diagnosis.
Recombinant His6-Bhlp29.7 was evaluated as a vaccine subunit for prevention of SD. The His6-Bhlp29.7 was shown to be immunogenic in mice following two intramuscular injections. Vaccination of mice with His6-Bhlp29.7 provided full protection after oral challenge with B. hyodysenteriae. In two experiments, intramuscular and oral vaccination of pigs with the His6-Bhlp29.7 resulted in a 50% reduction in incidence of SD compared to unvaccinated control pigs (P=0.047). This is the first subunit vaccine shown to provide pigs with protection from SD. Further work is needed to optimise delivery routes and adjuvants for commercial development of the vaccine.
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Idachaba, Stella Ejura. "Status of canine vaccination and the prevalence of rabies in humans and dogs in Plateau State, Nigeria 1998-2007." Pretoria : [s.n.], 2010. http://upetd.up.ac.za/thesis/available/etd-02252010-104653/.
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Schaten, Kathrin Maria. "One Health approach to measure the impact on wellbeing of selected infectious diseases in humans and animals in Zambia." Thesis, University of Edinburgh, 2018. http://hdl.handle.net/1842/33198.
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This study describes the results of a cross-sectional survey conducted in Mambwe district in the Eastern Province in Zambia. It uses a One Health approach to assess the impact of veterinary, medical, environmental and social determinants on animal and human health and wellbeing. One Health is defined as a holistic and interdisciplinary approach that describes the complexities between people, animals, the environment and their health. Human wellbeing is defined in this thesis as 'a condition in which all members of society are able to determine and meet their needs and have a large range of choices to meet their potential' (Prescott-Allen, 2001). As a first step, eight focus group discussions with the inhabitants followed by key informant interviews with stakeholders in the area were conducted to give a primary impression and narrow down the problems in relation to animal and human health of the area in general. Following this, a randomized selection of 210 households was visited and in each household blood samples were taken from all humans and all animals belonging to five animal species, namely cattle, goats, sheep, pigs and dogs. A third of the households did not keep any of the animal species chosen for sampling, but their inclusion was important for the social analysis. In all of these 210 households a wellbeing questionnaire was administered and, for every human and animal sampled, a health questionnaire. The study area falls within the tsetse-infested region of Zambia. It has a high wildlife density reflecting the proximity of several national parks and is historically endemic for both human and animal African trypanosomiasis (HAT&AAT). Therefore humans and animals were tested for trypanosomiasis using internal transcribed spacer (ITS) polymerase chain reaction (PCR). Since it is important as a differential diagnosis, malaria was tested for by a rapid diagnostic test in the field from human blood. Sera from mature individuals from all animal species except pigs were tested in a field laboratory for brucellosis using the Rose Bengal test. Additionally, cattle and dogs were tested for five genera of tick-borne infections (TBI) including Anaplasma, Ehrlichia, Theileria, Babesia and Rickettsia using reverse line blot (RLB) in the laboratory at the University of Edinburgh (UoE). The blood samples for PCR and RLB analysis at UoE were stored on WhatmanTM FTA cards. A total of 1012 human samples were tested for HAT and none found positive. 1005 (seven people had been tested positive or treated against malaria shortly before the sampling) people tested for malaria showed an overall prevalence of 15% (95% CI 13.2-17.7). None of the 734 Rose Bengal tests showed up positive for brucellosis. The prevalence of AAT in 1275 samples tested was much lower compared to former samplings; in cattle 22% (95% CI 18-27.2), in goats 7% (95% CI 4.5-9.2), in pigs 6% (95% CI 3.2-9.4), in dogs 9% (95% CI 5.2-13.6) and no samples were found positive in sheep. The prevalence of TBIs is much more complex with many multiple infections. A total of 340 cattle and 195 dogs were tested. In cattle the number of samples positive for any microorganism was as follows; 92% (95% CI 88- 94.2). Overall there were fewer positive samples from dogs with 25% of animals infected (95% CI 19.2-31.8). The wellbeing and health questionnaires were designed to help to identify possible risk factors for the above-mentioned diseases and signs, such as fever, diarrhoea and seizures, indicative for several other diseases. The results of these surveys might also help to identify potential reasons for a lower or higher prevalence of trypanosomiasis and malaria found than expected from previous studies. Additionally, information on personal happiness, attitudes towards veterinary and medical services, medical treatments received, education, women's reproductive history, drug abuse, people's perceptions of changes in environment and agriculture, demography, poverty and migration were collected via the questionnaires alongside information on livestock demographics and fertility. One of the main conclusions is that both medical and veterinary health care systems suffer from a number of shortcomings. The distance to appropriate treatment and care facilities is far and the necessary drugs are often unavailable. Also, both the knowledge and technology for diagnosing selected diseases is not in place. This study suggests that neurocysticercosis (NCC) plays an important role in this area due to the high number of seizures reported in people, in whom treatment for epilepsy was unsuccessful. Samples taken from a few pigs indicated the presence of Taenia solium, the causal agent of NCC. Furthermore, many of the TBIs are of zoonotic nature and further investigations must be made to begin to assess the burden of these diseases in humans and animals. Environmental changes such as degradation of the vegetation are likely to have an influence on the prevalence of studied diseases and this aspect is being investigated further in other studies. Due to the nature of a cross-sectional study, only limited conclusions can be drawn on the causal relationships of disease prevalence, but the social analysis conducted in this study confirmed the interactions of selected factors related to health and wealth unique for this study area.
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Bruce, Mieghan. "The impact of brucellosis in Albania : a systems approach." Thesis, Royal Veterinary College (University of London), 2016. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.701674.
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Viol, Milena Araúz [UNESP]. "Detecção de reações cruzadas por Leishmania spp. e Trypanosoma spp. em cães pelo ensaio imunoenzimático indireto, pela reação de imunofluorescência indireta e reação em cadeia de polimerase." Universidade Estadual Paulista (UNESP), 2011. http://hdl.handle.net/11449/94717.
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viol_ma_me_araca.pdf: 293015 bytes, checksum: bdc10c5bc3d8d20c951f2fc3b6dc7f4e (MD5)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
O objetivo deste estudo foi detectar reações cruzadas por Leishmania spp. e Trypanossoma cruzi pelo Ensaio Imunoenzimático Indireto (ELISA), pela Reação de Imunofluorescência Indireta (RIFI) e pela Reação em Cadeia da Polimerase (PCR). Assim, foram colhidas 408 amostras sanguíneas de cães domiciliados no município de Araçatuba,SP, de ambos os sexos, de diversas raças e com idade a partir de seis meses. Em relação à Leishmania spp., pela RIFI, 14,95 % (61/408) foram reagentes. A positividade por meio do ELISA, foi de 20,10% (82/408) e pela PCR, 29,66% (121/408), com diferença significativa para o sexo e a idade destes animais (p<0,05). Para Trypanosoma spp., a ocorrência de anticorpos pelo ELISA foi de 10,54% (43/408) e pela PCR, 2,45% (10/408) cães foram positivos. Pela RIFI, 10,29% (42/408) dos animais foram considerados positivos e somente o sexo apresentou diferença significativa (p<0,05). Neste trabalho, constatou-se que 10,54%(43/408) dos animais foram soropositivos por ELISA para Trypanosoma spp., sendo que 79,07%(34/43) obtiveram resultados positivos no diagnóstico molecular para Leishmania spp. e dos 10,29% (42/408) positivos por RIFI, 95,24% (40/42) dos cães confirmaram a infecção por este parasita. Por meio dos resultados obtidos, pode-se inferir que foram evidenciadas reações cruzadas nos ensaios sorológicos para ambos os protozoários, nos animais analisados neste trabalho
The aim of this study was to detect cross-infection by Leishmania spp. and Trypanosoma spp. by indirect immunosorbent assay (ELISA), by Indirect Immunofluorescence (IFA) and by the Polymerase Chain Reaction (PCR). Thus, blood samples were collected from 408 domestic dogs of both sexes, different races and ages from six months. For Leishmania spp. by IFA, 14.95% (61/408) were positive. Positive by ELISA, was 20.10% (82/408), and PCR 29.66% (121/408), with significant difference for sex and age of animals (p <0.05). For Trypanosoma spp., the occurrence of antibodies by ELISA, was 10.54% (43/408), and PCR, 2.45% (10/408) dogs were positive. By IFA, 10.29% (42/408) of the animals were considered positive and only sex was significant difference (p <0.05). In this work it was found that 10.54% (43/408) animals were seropositive by ELISA for Trypanosoma spp., 79.07% (34/43) had positive results in molecular diagnostic for Leishmania spp. and 10.29% (42/408) positive by IFA, 95.24% (40/42) dogs confirmed the infection by this parasite. Through the results obtained can be inferred that cross-infection were observed by both protozoa in animals of this paper
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Modumo, Jacob. "Determination of the minimum protective dose for bluetongue serotype 2, 4 and 8 vaccines in sheep." Diss., Pretoria : [s.n.], 2009. http://upetd.up.ac.za/thesis/available/etd-07312009-145842/.
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Rodrigues, Acácio Romoaldo Assoni [UNESP]. "Programa de erradicação da febre aftosa no estado de São Paulo: avaliação dos dados oficiais obtidos no período de 1997 a 2012." Universidade Estadual Paulista (UNESP), 2014. http://hdl.handle.net/11449/121911.
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000815117.pdf: 1605584 bytes, checksum: 806edb3db344e3acd66cec8448e8efae (MD5)A febre aftosa (FA), doença altamente contagiosa que afeta os animais biungulados, é reconhecida pelos prejuízos econômicos nas propriedades e nas regiões onde é endêmica. O Estado de São Paulo não registra focos de FA desde 1996. A partir de dados oficiais obtidos de 1997 a 2012, este estudo organizou, analisou e confrontou com a literatura pertinente as taxas de vacinação contra FA nas diferentes etapas e nas diversas regiões do Estado de São Paulo; analisou os resultados do monitoramento oficial delineado para demonstrar a eficiência da vacinação contra a FA; analisou os procedimentos de vigilância passiva e de atendimento às notificações de suspeitas de doenças vesiculares no Estado de São Paulo; analisou os resultados do monitoramento oficial delineado para demonstrar a ausência de circulação viral no Estado de São Paulo; relacionou as medidas preventivas adotadas pela Coordenadoria de Defesa Agropecuária, da Secretaria de Agricultura e Abastecimento do Estado de São Paulo (CDA/SAA/SP) diante de focos de FA em outros Estados limítrofes. Finalmente, foram descritas as perspectivas de implementação de área livre sem vacinação para o Estado de São Paulo, classificação outorgada pela Organização Mundial de Saúde Animal (OIE)
Foot-and-mouth disease (FMD), a highly contagious disease that affects cloven-hoofed animals, is recognized by economic losses on properties and in the regions where it is endemic. The State of São Paulo has had no records of outbreaks of FMD since 1996. From official data from 1997 to 2012, this study organized, analyzed and confronted with relevant literature rates of FMD vaccination at different periods and in different regions of the State of São Paulo; analyzed the results of the official monitoring designed to demonstrate the efficiency of vaccination against FMD; examined the passive surveillance procedures and care to reports of suspected vesicular disease in the State of São Paulo; analysed the results of the official monitoring designed to demonstrate the absence of viral circulation in the State of São Paulo; related preventive measures adopted by Coordenadoria de Defesa Agropecuária, the Secretaria de Agricultura e Abastecimento of the Estado de São Paulo before outbreaks of FMD in other neighboring States. Finally, the prospects of implementing free zone without vaccination for the State of São Paulo, ranking granted by the World Organization for Animal Health
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Rodrigues, Acácio Romoaldo Assoni. "Programa de erradicação da febre aftosa no estado de São Paulo : avaliação dos dados oficiais obtidos no período de 1997 a 2012 /." Jaboticabal, 2014. http://hdl.handle.net/11449/121911.
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Orientador: Samir Issa SamaraCoorientador: Luis Guilherme de Oliveira
Banca: Fernando Gomes Buchala
Banca: Aramis Augusto Pinto
Resumo: A febre aftosa (FA), doença altamente contagiosa que afeta os animais biungulados, é reconhecida pelos prejuízos econômicos nas propriedades e nas regiões onde é endêmica. O Estado de São Paulo não registra focos de FA desde 1996. A partir de dados oficiais obtidos de 1997 a 2012, este estudo organizou, analisou e confrontou com a literatura pertinente as taxas de vacinação contra FA nas diferentes etapas e nas diversas regiões do Estado de São Paulo; analisou os resultados do monitoramento oficial delineado para demonstrar a eficiência da vacinação contra a FA; analisou os procedimentos de vigilância passiva e de atendimento às notificações de suspeitas de doenças vesiculares no Estado de São Paulo; analisou os resultados do monitoramento oficial delineado para demonstrar a ausência de circulação viral no Estado de São Paulo; relacionou as medidas preventivas adotadas pela Coordenadoria de Defesa Agropecuária, da Secretaria de Agricultura e Abastecimento do Estado de São Paulo (CDA/SAA/SP) diante de focos de FA em outros Estados limítrofes. Finalmente, foram descritas as perspectivas de implementação de área livre sem vacinação para o Estado de São Paulo, classificação outorgada pela Organização Mundial de Saúde Animal (OIE)
Abstract: Foot-and-mouth disease (FMD), a highly contagious disease that affects cloven-hoofed animals, is recognized by economic losses on properties and in the regions where it is endemic. The State of São Paulo has had no records of outbreaks of FMD since 1996. From official data from 1997 to 2012, this study organized, analyzed and confronted with relevant literature rates of FMD vaccination at different periods and in different regions of the State of São Paulo; analyzed the results of the official monitoring designed to demonstrate the efficiency of vaccination against FMD; examined the passive surveillance procedures and care to reports of suspected vesicular disease in the State of São Paulo; analysed the results of the official monitoring designed to demonstrate the absence of viral circulation in the State of São Paulo; related preventive measures adopted by Coordenadoria de Defesa Agropecuária, the Secretaria de Agricultura e Abastecimento of the Estado de São Paulo before outbreaks of FMD in other neighboring States. Finally, the prospects of implementing free zone without vaccination for the State of São Paulo, ranking granted by the World Organization for Animal Health
Mestre
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Viol, Milena Araúz. "Detecção de reações cruzadas por Leishmania spp. e Trypanosoma spp. em cães pelo ensaio imunoenzimático indireto, pela reação de imunofluorescência indireta e reação em cadeia de polimerase /." Araçatuba : [s.n.], 2011. http://hdl.handle.net/11449/94717.
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Orientador: Katia Denise Saraiva BrescianiBanca: Renato Andreotti e Silva
Banca: Valéria Marçal Felix de Lima
Resumo: O objetivo deste estudo foi detectar reações cruzadas por Leishmania spp. e Trypanossoma cruzi pelo Ensaio Imunoenzimático Indireto (ELISA), pela Reação de Imunofluorescência Indireta (RIFI) e pela Reação em Cadeia da Polimerase (PCR). Assim, foram colhidas 408 amostras sanguíneas de cães domiciliados no município de Araçatuba,SP, de ambos os sexos, de diversas raças e com idade a partir de seis meses. Em relação à Leishmania spp., pela RIFI, 14,95 % (61/408) foram reagentes. A positividade por meio do ELISA, foi de 20,10% (82/408) e pela PCR, 29,66% (121/408), com diferença significativa para o sexo e a idade destes animais (p<0,05). Para Trypanosoma spp., a ocorrência de anticorpos pelo ELISA foi de 10,54% (43/408) e pela PCR, 2,45% (10/408) cães foram positivos. Pela RIFI, 10,29% (42/408) dos animais foram considerados positivos e somente o sexo apresentou diferença significativa (p<0,05). Neste trabalho, constatou-se que 10,54%(43/408) dos animais foram soropositivos por ELISA para Trypanosoma spp., sendo que 79,07%(34/43) obtiveram resultados positivos no diagnóstico molecular para Leishmania spp. e dos 10,29% (42/408) positivos por RIFI, 95,24% (40/42) dos cães confirmaram a infecção por este parasita. Por meio dos resultados obtidos, pode-se inferir que foram evidenciadas reações cruzadas nos ensaios sorológicos para ambos os protozoários, nos animais analisados neste trabalho
Abstract: The aim of this study was to detect cross-infection by Leishmania spp. and Trypanosoma spp. by indirect immunosorbent assay (ELISA), by Indirect Immunofluorescence (IFA) and by the Polymerase Chain Reaction (PCR). Thus, blood samples were collected from 408 domestic dogs of both sexes, different races and ages from six months. For Leishmania spp. by IFA, 14.95% (61/408) were positive. Positive by ELISA, was 20.10% (82/408), and PCR 29.66% (121/408), with significant difference for sex and age of animals (p <0.05). For Trypanosoma spp., the occurrence of antibodies by ELISA, was 10.54% (43/408), and PCR, 2.45% (10/408) dogs were positive. By IFA, 10.29% (42/408) of the animals were considered positive and only sex was significant difference (p <0.05). In this work it was found that 10.54% (43/408) animals were seropositive by ELISA for Trypanosoma spp., 79.07% (34/43) had positive results in molecular diagnostic for Leishmania spp. and 10.29% (42/408) positive by IFA, 95.24% (40/42) dogs confirmed the infection by this parasite. Through the results obtained can be inferred that cross-infection were observed by both protozoa in animals of this paper
Mestre
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Weyer, Jacqueline. "Immune responses against recombinant poxvirus vaccines that express full-length lyssavirus glycoprotein genes." Thesis, University of Pretoria, 2006. http://hdl.handle.net/2263/28113.
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Rabies is a fatal but preventable neurotropic disease of potentially all mammals. The disease is caused by lyssaviruses. Rabies is recognized as the 10th most common lethal infectious disease in the world, rendering it one of the most feared zoonotic diseases known to man. Nevertheless, rabies can be prevented by application of pre- or post exposure treatments. Rabies vaccines have been available since the time of Pasteur, more that one hundred years ago. Since, vaccine research focused on the development of safer and more effective vaccines. Topics of current interest in the field of rabies vaccinology were addressed in this study. A primary concern regarding the disease is human mortalities, in the range of 60 000, reported every year. Most of these are linked to exposure to rabid dogs. In addition, a great number of post exposure treatments are administered each year at great costs. Despite availability of efficacious biologics, several factors influence the optimal use and accessibility of these agents in the countries of interest, with cost and availability being the major contributing factors. A proven approach is mass oral vaccination of target animals, such as dogs, which indirectly infers protection to susceptible hosts, including man. Currently available vaccines present several disadvantages of use though, including issues of safety or doubtful stability. Safer but effective alternative vaccines that could be used in oral baits would be valuable. Here the use of two candidate host restricted poxvirus vaccine vectors were explored, particularly also in regard to oral innocuity. The construction, convenient isolation and use of a recombinant Lumpy skin disease virus (Neethling strain) expressing rabies virus glycoprotein in a mouse model were investigated. In addition, a recombinant Modified Vaccinia virus Ankara expressing rabies virus glycoprotein was prepared and tested as a vaccine in mice, dogs and raccoons. In both cases it was clear that the severe attenuation of these viruses did affect the efficacy of the recombinant vaccines in the non-permissive hosts. With the recombinant MVA a clear dosage effect could be shown, and equivalent humoral responses could only be attained at much higher titers of vaccine virus as with replication competent counterparts. Secondly, the cross-protection of rabies vaccines across the spectrum of lyssaviruses was addressed. Lyssaviruses can be divided into two groups based on sequence analysis and pathogenesis. Viruses belonging to the so-called phylogroup II, are the Mokola, Lagos and West Caucasian Bat viruses. Classic rabies biologics fail to fully protect against the viruses attributed to a lack of cross-neutralization. Here, cross-protection and cross-reactive immune responses induced by recombinant vaccinia viruses expressing rabies, Mokola or West Caucasian Bat virus glycoproteins, in single or dual combinations, were investigated. As expected, there was a lack of cross-protection of rabies and Mokola glycoprotein vaccines. There was also a clear lack of cross-protection of West Caucasian Bat virus glycoprotein vaccine and rabies and Mokola viruses. The dual antigen expressing vaccines did not appear to offer any additional protective effect in the tested model. The Mokola virus glycoprotein vaccines induced neutralizing antibody responses that significantly cross-neutralized Lagos Bat virus.Thesis (PhD (MIcrobiology))--University of Pretoria, 2007.
Microbiology and Plant Pathology
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Babboni, Selene Daniela [UNESP]. "Compêndio para planejamento de campanha de vacinação anual contra a raiva de cães e gatos na zona urbana de municípios." Universidade Estadual Paulista (UNESP), 2015. http://hdl.handle.net/11449/140241.
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000866849.pdf: 1100440 bytes, checksum: 1e6bed6dfed6644cd4635ccdb9bd4b7e (MD5)A Raiva continua sendo uma enfermidade de preocupação mundial com severidade de ser praticamente 100% letal para as pessoas tendo o cão como seu principal transmissor. O procedimento mais eficaz para controlar a enfermidade é quando são vacinados 75% dos cães de um Município, quantidade que contribui para a interrupção do elo epidemiológico de transmissão da doença e, por consequência, proteção à saúde pública. A experiência acumulada desde 1968 em planejamentos de campanhas de vacinação anual em cães e gatos em diferentes Municípios do estado de São Paulo objetivou a elaboração do presente compêndio. Os materiais e métodos foram descritos e analisados sobre aspectos temáticos e operacionais de áreas do conhecimento sendo empreendidos como procedimentos para permitir sustentação para elaborações de políticas setoriais subsequentes. O desenvolvimento do compêndio, portanto, foi sustentado nesses 47 anos de planejamentos de campanhas de vacinação anual contra a Raiva baseando-se principalmente nas características da área e nas dinâmicas populacionais animal e humana dos Municípios alvos das campanhas, detalhando os procedimentos operacionais necessários para que a meta de vacinação dos animais possa ser atingida em cada Município, considerando-se ainda as condições socioeconômicas e culturais das áreas objetos dos planejamentos das campanhas de vacinação
Rabies is still a concerning worldwide disease with almost 100% lethality to people and the dog acting as the main carrier. The most efficient proceeding to control the disease is when 75% of the dog population in one city is vaccinated, amount that contributes to the interruption of the disease epidemiological transmission cycle, and, by consequence, public health protection. The experience accumulated since 1968 in planning the annual vaccination campaign in dogs and cats in different cities from São Paulo state aimed the elaboration of the present compendium. Material and methods were described and analyzed under the thematic and operational aspects from areas of knowledge and undertaken as proceedings to allow sustaining to elaborate subsequential sectorial politics. The compendium development, thus, was sustained on these 47 years of annual vaccination campaign against rabies based mainly on the area characteristics and animal and human populational dynamics at the target cities detailing the operational proceedings needed so that the goal to vaccinate the animals be achieved in each city considering further the cultural and socioeconomic conditions of the areas from the vaccination campaign planning
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Babboni, Selene Daniela. "Compêndio para planejamento de campanha de vacinação anual contra a raiva de cães e gatos na zona urbana de municípios." Botucatu, 2015. http://hdl.handle.net/11449/140241.
Full textAbstract:
Orientador: José Rafael ModoloBanca: Denise Lopes
Banca: José Paes de Almeida Nogueira Pinto
Banca: Hení Falcão da Costa
Banca: João Marcelo Azevedo de Paula Antunes
Resumo: A Raiva continua sendo uma enfermidade de preocupação mundial com severidade de ser praticamente 100% letal para as pessoas tendo o cão como seu principal transmissor. O procedimento mais eficaz para controlar a enfermidade é quando são vacinados 75% dos cães de um Município, quantidade que contribui para a interrupção do elo epidemiológico de transmissão da doença e, por consequência, proteção à saúde pública. A experiência acumulada desde 1968 em planejamentos de campanhas de vacinação anual em cães e gatos em diferentes Municípios do estado de São Paulo objetivou a elaboração do presente compêndio. Os materiais e métodos foram descritos e analisados sobre aspectos temáticos e operacionais de áreas do conhecimento sendo empreendidos como procedimentos para permitir sustentação para elaborações de políticas setoriais subsequentes. O desenvolvimento do compêndio, portanto, foi sustentado nesses 47 anos de planejamentos de campanhas de vacinação anual contra a Raiva baseando-se principalmente nas características da área e nas dinâmicas populacionais animal e humana dos Municípios alvos das campanhas, detalhando os procedimentos operacionais necessários para que a meta de vacinação dos animais possa ser atingida em cada Município, considerando-se ainda as condições socioeconômicas e culturais das áreas objetos dos planejamentos das campanhas de vacinação
Abstract: Rabies is still a concerning worldwide disease with almost 100% lethality to people and the dog acting as the main carrier. The most efficient proceeding to control the disease is when 75% of the dog population in one city is vaccinated, amount that contributes to the interruption of the disease epidemiological transmission cycle, and, by consequence, public health protection. The experience accumulated since 1968 in planning the annual vaccination campaign in dogs and cats in different cities from São Paulo state aimed the elaboration of the present compendium. Material and methods were described and analyzed under the thematic and operational aspects from areas of knowledge and undertaken as proceedings to allow sustaining to elaborate subsequential sectorial politics. The compendium development, thus, was sustained on these 47 years of annual vaccination campaign against rabies based mainly on the area characteristics and animal and human populational dynamics at the target cities detailing the operational proceedings needed so that the goal to vaccinate the animals be achieved in each city considering further the cultural and socioeconomic conditions of the areas from the vaccination campaign planning
Doutor
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Camossi, Lucilene Granuzzio [UNESP]. "Vacina irradiada de Toxoplasma gondii em rats wistar: análise molecular da carga parasitária tecidual, transmissão congênita e eliminação do agente pelo leite." Universidade Estadual Paulista (UNESP), 2013. http://hdl.handle.net/11449/103806.
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000738028.pdf: 1911096 bytes, checksum: 107327a17db129713052a243c7622095 (MD5)A toxoplasmose se destaca pela elevada importância médica, veterinária e zootécnica, e seu controle tornou-se um desafio para a saúde pública em todo o mundo, sinalizando a prioridade em buscar na vacinação uma estratégia para conter a infecção. Este estudo foi realizado com o objetivo de avaliar a proteção conferida pela pré-imunização, com vacina constituída de taquizoítos irradiados a 255 Gy, com raios γ, em ratas Wistar, contra desafios orais por diferentes estágios infectantes do Toxoplasma gondii. Nos experimentos foi investigada a proteção vacinal contra formação de cistos cerebrais, cistos na musculatura esquelética, transmissão congênita e eliminação do agente pelo leite. A vacina foi administrada por via oral, com uso de adjuvante, em três doses com intervalos de 15 dias, antes do acasalamento das ratas. Constatada a gestação, realizou-se o desafio oral das ratas com bradizoítos, oocistos e taquizoítos de T. gondii. Foram utilizadas ratas Wistar em idade reprodutiva, que foram distribuídas em dois experimentos, composto por oito grupos cada. No experimento I cada grupo foi formado por seis ratas e, no experimento II, por cinco ratas. Os animais em cada experimento foram distribuídos da seguinte forma: Grupo 1: imunizado e desafiado com bradizoítos (ME-49) de T. gondii; Grupo 2: desafiado com bradizoítos (ME-49) de T. gondii; Grupo 3: imunizado e desafiado com oocistos (ME-49) de T. gondii; Grupo 4: desafiado com oocistos (ME-49) de T. gondii; Grupo 5: imunizado e desafiado com taquizoítos (RH) de T. gondii; Grupo 6: desafiado com taquizoítos (RH) de T. gondii; Grupo 7: apenas imunizado, Grupo 8: não imunizado e não desafiado. Semanalmente, desde a imunização, foram realizadas análises sorológicas e hematológicas. Após a parição (experimento I), as ratas e neonatos foram eutanasiados para determinação da carga parasitária no cérebro e na musculatura esquelética...
Toxoplasmosis has high medical and veterinary importance, their control has become a challenge to public health worldwide, signaling the priority in seeking vaccination strategy to contain the infection.The objective of the present study was to evaluate the efficacy of Toxoplasma gondii (T. gondii) tachyzoites irradiated vaccine, administered in experimentally inoculated Wistar female rats by oral challenges against different infectious stage of Toxoplasma gondii for blocking the production of T. gondii cysts in brain and muscle tissues, congenital transmission and also the elimination of the parasite in milk. Vaccine was administered orally, plus aluminium hidroxy as adjuvant, with three doses of 15 days interval. One week after pregnancy confirmation, challenge was done by gavage with T. gondii cysts, oocysts and tachizoites. Wistar rats in reproductive stage were distributed in two studies, composed by eight groups of six animals each (experiment I) an five animals each (experiment II), as follows: Group 1: immunized and challenge with bradizoites of ME-49 T. gondii strain; Group 2: non- immunized and challenged bradizoites of ME-49 T. gondii strain; Group 3: immunized and challenged with oocysts of ME-49 T. gondii strain; Group 4: non- immunized and challenged with oocysts of ME-49 T. gondii strain; Group 5: immunized and challenged with tachizoites of RH T. gondii strain; Group 6: non- immunized and challenged with tachizoites of RH T. gondii strain; Group 7: only immunized, Group 8: non- immunized and non-challenged. Weekly, after vaccination serological and hematological were performed. After parturition (Experiment I), the rats and newborns were euthanized, tissues and organs were collected for parasite load evaluation by Real Time PCR, intestinal tissue was collected for subsequent histological examinations, immunological involved in immune response against T. gondii. During lactation ...
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Camossi, Lucilene Granuzzio. "Vacina irradiada de Toxoplasma gondii em rats wistar : análise molecular da carga parasitária tecidual, transmissão congênita e eliminação do agente pelo leite /." Botucatu, 2013. http://hdl.handle.net/11449/103806.
Full textAbstract:
Orientador: Helio LangoniBanca: Italmar Teodorico Navarro
Banca: Vírginia Bodelão Richini
Banca: Simone Bandini Lucheis
Banca: Rodrigo Costa da Silva
Resumo: A toxoplasmose se destaca pela elevada importância médica, veterinária e zootécnica, e seu controle tornou-se um desafio para a saúde pública em todo o mundo, sinalizando a prioridade em buscar na vacinação uma estratégia para conter a infecção. Este estudo foi realizado com o objetivo de avaliar a proteção conferida pela pré-imunização, com vacina constituída de taquizoítos irradiados a 255 Gy, com raios γ, em ratas Wistar, contra desafios orais por diferentes estágios infectantes do Toxoplasma gondii. Nos experimentos foi investigada a proteção vacinal contra formação de cistos cerebrais, cistos na musculatura esquelética, transmissão congênita e eliminação do agente pelo leite. A vacina foi administrada por via oral, com uso de adjuvante, em três doses com intervalos de 15 dias, antes do acasalamento das ratas. Constatada a gestação, realizou-se o desafio oral das ratas com bradizoítos, oocistos e taquizoítos de T. gondii. Foram utilizadas ratas Wistar em idade reprodutiva, que foram distribuídas em dois experimentos, composto por oito grupos cada. No experimento I cada grupo foi formado por seis ratas e, no experimento II, por cinco ratas. Os animais em cada experimento foram distribuídos da seguinte forma: Grupo 1: imunizado e desafiado com bradizoítos (ME-49) de T. gondii; Grupo 2: desafiado com bradizoítos (ME-49) de T. gondii; Grupo 3: imunizado e desafiado com oocistos (ME-49) de T. gondii; Grupo 4: desafiado com oocistos (ME-49) de T. gondii; Grupo 5: imunizado e desafiado com taquizoítos (RH) de T. gondii; Grupo 6: desafiado com taquizoítos (RH) de T. gondii; Grupo 7: apenas imunizado, Grupo 8: não imunizado e não desafiado. Semanalmente, desde a imunização, foram realizadas análises sorológicas e hematológicas. Após a parição (experimento I), as ratas e neonatos foram eutanasiados para determinação da carga parasitária no cérebro e na musculatura esquelética ...
Abstract: Toxoplasmosis has high medical and veterinary importance, their control has become a challenge to public health worldwide, signaling the priority in seeking vaccination strategy to contain the infection.The objective of the present study was to evaluate the efficacy of Toxoplasma gondii (T. gondii) tachyzoites irradiated vaccine, administered in experimentally inoculated Wistar female rats by oral challenges against different infectious stage of Toxoplasma gondii for blocking the production of T. gondii cysts in brain and muscle tissues, congenital transmission and also the elimination of the parasite in milk. Vaccine was administered orally, plus aluminium hidroxy as adjuvant, with three doses of 15 days interval. One week after pregnancy confirmation, challenge was done by gavage with T. gondii cysts, oocysts and tachizoites. Wistar rats in reproductive stage were distributed in two studies, composed by eight groups of six animals each (experiment I) an five animals each (experiment II), as follows: Group 1: immunized and challenge with bradizoites of ME-49 T. gondii strain; Group 2: non- immunized and challenged bradizoites of ME-49 T. gondii strain; Group 3: immunized and challenged with oocysts of ME-49 T. gondii strain; Group 4: non- immunized and challenged with oocysts of ME-49 T. gondii strain; Group 5: immunized and challenged with tachizoites of RH T. gondii strain; Group 6: non- immunized and challenged with tachizoites of RH T. gondii strain; Group 7: only immunized, Group 8: non- immunized and non-challenged. Weekly, after vaccination serological and hematological were performed. After parturition (Experiment I), the rats and newborns were euthanized, tissues and organs were collected for parasite load evaluation by Real Time PCR, intestinal tissue was collected for subsequent histological examinations, immunological involved in immune response against T. gondii. During lactation ...
Doutor
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Klimovich, Charlotte Marie. "Investigations into the physiological and biomechanical basis of differential success in oral rabies vaccination between skunks (Mephitis mephitis) and raccoons (Procyon lotor)." Ohio University Honors Tutorial College / OhioLINK, 2017. http://rave.ohiolink.edu/etdc/view?acc_num=ouhonors1502454684921845.
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Mjihdi, Abdelkarim. "Capacité de reproduction de la souris et infection aiguë par Trypanosoma cruzi." Doctoral thesis, Universite Libre de Bruxelles, 2004. http://hdl.handle.net/2013/ULB-DIPOT:oai:dipot.ulb.ac.be:2013/211065.
Full textAbstract:
Trypanosoma cruzi est un parasite protozoaire à multiplication intracellulaire, agent de la maladie de Chagas, infectant 16 à 18 millions de personnes en Amérique latine. Il peut être transmis de la mère infectée au fœtus dans 2 à 10 % des cas, mais ses autres effets sur la gestation ont été peu étudiés. Par ailleurs, les cytokines ont des effets sur la gestation. Certaines d’entre elles, comme l’interleukine-1, l’IL-4, l’IL-5, l’IL-10, le GM-CSF et le TGF-b2, sont bénéfiques pour la gestation, tandis que d’autres, comme l’IL-2, l’IL-12, l’IFN-g et le TNF-a ont des effets nocifs sur celle-ci. L’impact de l’infection à T. cruzi, stimulant la production de TNF-a et d’IFN-g, sur l'implantation et la croissance fœtale n’a pas été étudié.Le but de notre travail était d’étudier les effets de l’infection aiguë à T. cruzi sur la capacité de reproduction de la souris. Nous avons ainsi évalué les effets de cette infection sur la fertilité, le développement et la viabilité des fœtus de souris et le rôle de l’IFN-g et du TNF produits au cours de l’infection sur le développement de la gestation.
Nous avons montré que l’infection aiguë à T. cruzi :i) diminue la capacité de reproduction de la souris ;ii) provoque une mortalité fœtale massive précoce (résorptions), tardive et néonatale associée à un retard de croissance intra-utérin, et ce, iii) en dehors de toute transmission congénitale du parasite.
Par ailleurs nos travaux montrent que la mortalité fœtale/néonatale est associée à une invasion parasitaire massive du placenta qui présente d’importantes lésions à type d’infiltrats inflammatoires, de nécrose ischémique, de dépôts de fibrine et de thromboses vasculaires. Nous avons noté qu’il existe une relation inverse entre la charge parasitaire des unités utéro-placentaires et la viabilité du conceptus, suggérant que ces lésions placentaires contribuent à la mortalité fœtale en limitant les échanges materno-fœtaux.
Enfin, nous avons également étudié le rôle de cytokines abortogènes comme le TNF et l’IFN-g, produites abondamment pendant l’infection aiguë de la souris par T. cruzi. Les taux sanguins maternels d’IFN-g étaient augmentés au 9ième mais pas aux 17ième et 19ième jours de gestation, alors que les taux de TNF sanguin et la production placentaire de cette cytokine augmentaient aux 17ième et 19ième jours de gestation. Afin d’évaluer le rôle de ces deux cytokines dans la mortalité fœtale, des souris ont été traitées par la pentoxifylline, pour inhiber la transcription du gène de TNF-a et diminuer la production d’IFN-g. Ces souris montraient une réduction de la mortalité fœtale à mi-gestation, associée à une diminution de la production du TNF placentaire, sans modifications des taux systémiques et sans effets sur l’IFN-g, suggérant la contribution du TNF dans la mortalité fœtale associée à l’infection aiguë par T. cruzi.
En conclusion, notre travail montre que l’infection aiguë à Trypanosoma cruzi exerce un effet particulièrement néfaste sur la capacité de reproduction et le développement de la gestation chez la souris et que les lésions placentaires liées à l’infection et la production de TNF par le placenta infecté contribuent à cet effet.
Doctorat en sciences biomédicales
info:eu-repo/semantics/nonPublished
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Eyssen, Lauren Elizabeth-Ann. "Studying trypanosomal peptidase antigen targets for the diagnosis of animal African trypanosomiasis." Thesis, 2013. http://hdl.handle.net/10413/11165.
Full textAbstract:
The lack of a vaccine candidate due to antigenic variation by trypanosomal parasites, the causative agents of human and animal African trypanosomiasis, requires the disease to be controlled by surveillance, diagnosis and appropriate treatment schedules. Due to the non-specific symptoms along with the toxicity and side effects of the current trypanocides, diagnosis needs to be accurate, cost effective and applicable to active case finding in mostly rural settings. Trypanosomal proteases have been identified as virulence factors as they are essential to the parasites‟ survival. Here the diagnostic potential of previously described virulence factors, oligopeptidase B (OPB), pyroglutamyl peptidase (PGP) and the full length and catalytic domain of the cathepsin L-like peptidases (CATLFL and CATL respectively) from T. congolense (Tc) as well as OPB and CATL from T. vivax (Tv), was determined. These antigens were recombinantly expressed, purified and used to generate antibodies in chickens. The purified recombinant antigens were tested in an inhibition and indirect ELISA format using two separate blinded serum panels consisting of sera from non-infected and experimentally infected cattle, one each for T. congolense and for T. vivax. The tested sera were diluted 1:10 for the TcCATLFL, TcCATL antigens whilst the TvCATL antigen used a 1:100 serum dilution. The TcCATLFL, TcCATL and TvCATL antigens had the highest diagnostic potential in the indirect ELISA format with a 90.91, 92.21% accuracy at the second cut-off and a 77.22% accuracy at the third cut-off along with 0.8084, 0.7785 and 0.8813 area under curve (AUC) values respectively. These antigens show potential for development of lateral flow tests to detect T. congolense and T. vivax infections in cattle. The recently discovered metacaspases (MCAs) have been implicated in caspase-like activity and differentiation in T. b. brucei, T. cruzi and L. major and are considered to be virulence factors. The putative metacaspase 5 gene from T. congolense (TcMCA5) was successfully cloned, expressed within inclusion bodies, resolubilised and refolded using immobilised metal affinity chromatography. Recombinant TcMCA5 was successfully refolded as evident by the hydrolysis of the synthetic peptide substrate, Z-Gly-Gly-Arg-AMC. Autocatalytic processing was observed within the inclusion bodies and the products were purified along with the full length recombinant protein. Anti-TcMCA5 IgY antibodies, raised in chickens, were able to detect the native TcMCA5 along with the autocatalytic processed products within the lysate of the procyclic T. congolense (strain IL 3000) parasites. The diagnostic potential of TcMCA5 still requires verification.Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2013.
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Bartlett, Cara-Lesley. "Recombinant expression and evaluation of a- and b- tubulin from Trypanosoma congolense as vaccine candidates for African trypanosomiasis." Thesis, 2010. http://hdl.handle.net/10413/4768.
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African trypanosomiasis is caused by protozoan parasites known as trypanosomes, which are transmitted by the tsetse fly, affecting both humans and animals. Trypanosoma congolense is one of the main trypanosome species affecting cattle and causes the disease known as nagana. Control of animal African trypanosomiasis currently relies on
chemotherapy and vector control methods, neither of which has proven satisfactory. An effective vaccine against trypanosomiasis would be the most cost effective solution to control the disease; however, due to the phenomenon of antigenic variation, intrinsic to the parasite’s outer coat of variable surface glycoprotein, this has not yet been achieved. Recent
vaccine efforts have been centred on identification of invariant parasite antigens for use as vaccine candidates. Trypanosome cytoskeleton components have in recent years been shown to be capable of providing a protective immune response against trypanosome infection. These include
tubulin proteins, which form the main components of the cytoskeleton, as well as microtubule associated proteins (MAPs) and paraflagellar rod proteins. In the present study α- and β-tubulin from T. congolense were recombinantly expressed and their immuno-protective potential in mice assessed. Amplification of both α- and β-tubulin ORFs from T. congolense genomic DNA was followed by cloning of the amplicons into the T-vector pTZ57R/T, and thereafter sub-cloning into the bacterial expression vector,
pET238a and the yeast expression vector pPICZαA28. Only the α-tubulin amplicon was successfully sub-cloned into pICZAαA28; however, no protein expression was achieved upon transfection of the methylotrophic yeast, Pichia pastoris, with this construct. Subcloning of both α- and β-tubulin inserts into pET28a was successful. Expression of recombinant α- and β-tubulin as fusion proteins with a histidine tag, both at a size of 55 kDa, was achieved in Escherichia coli host BL21 (DE3). Recombinant proteins were successfully purified using nickel chelate chromatography under denaturing conditions. Refolding was first attempted by dilution of purified
denatured proteins in a refolding buffer followed by reconcentration, but was largely unsuccessful. A second, more successful refolding method was performed wherein denatured proteins were refolded by application of a decreasing gradient of urea, while bound to a nickel chelate column. Native tubulin from cultured T.congolense procyclics was successfully purified and renatured using a polymerisation/depolymerisation method for use as a control for immunisation. Mice were immunised separately with refolded recombinant α- and β-tubulin, native tubulin or an irrelevant protein VP4AA expressed in the same way as the tubulins. ELISA analysis confirmed the production of antibodies against each protein. Parasitaemia developed in all mice following challenge with T. congolense. Only the group immunised
with β-tubulin recorded no deaths during the monitoring period despite the presence of parasitaemia, with 60% of mice immunised with α-tubulin or VP4AA and the no antigen control and no mice from the native tubulin immunised group surviving. The results showed that partial protection against trypanosomiasis caused by T. congolense infection was achieved in the group immunised with β-tubulin and suggest that β-tubulin may have
vaccine potential.Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2010.
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Kalundi, Erastus Mulinge. "Molecular analysis of the congopain gene family." Thesis, 2008. http://hdl.handle.net/10413/4140.
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Animal trypanosomosis is a major constraint in livestock production in Sub-Saharan Africa. With the emergence of resistance against trypanocidal drugs, the cost and environmental concerns raised by vector control, and the challenge of antigenic variation in vaccine development, alternative control measures are being sought. An anti-disease strategy, whereby the immune response or chemotherapy is aimed towards pathogenic factors rather than the parasite itself, constitutes such a novel approach. Congopain is the major cysteine protease in Trypanosoma congolense, and upon release in the bloodstream of infected cattle, acts as a pathogenic factor. It is therefore an attractive candidate for an anti-disease vaccine. It was hence deemed necessary to investigate the variability of congopain-like cysteine proteases before attempting to design drugs and vaccines based on the inhibition of congopain. Most congopain-like cysteine protease genes of T. congolense exist in a single locus of 12-14 copies organised as tandem repeats of 2 kb gene units. A gene unit library of 120 clones was constructed out of several cosmid clones selected in a previous study that contained various lengths of the congopain locus. Some 24 gene unit clones were sequenced, and it was found that congopain genes cluster in three sub-families, named CP1 (8 clones), CP2 (12 clones) and CP3 (4 clones). The latter most characteristically shows a substitution of the active site cysteine by a serine. Isoform specific primers were designed and used to verify the proportions of the three isoforms (one third CP1, half CP2 and a sixth CP3) in the remaining clones of the library. Since this first study was conducted in one isolate, IL 3000, the results were subsequently validated in a large array of isolates, of T. congolense, as well as T. vivax and T. brucei subspecies, by a PCR approach. Finally, to gain access to copies of congopain genes that are not present in the locus, but rather scattered in the genome, an attempt was made to construct a 2 kb size-restricted genomic library. Only 206 clones could be produced, of which a mere 8 coded for congopain-like proteases. The fact that 7 out of 8 of these clones belong to CP3 (thought to be inactive) suggested a cloning artefact, possibly related to the activity of the cloned proteases.
Overall, all congopain genes appear very conserved in a given species, with 87-99% identity at protein level. The pre- and pro-region were the most conserved, while the catalytic domain was the most variable, especially around the active site cysteine, with frequent replacement by a serine residue, and in one instance by phenylalanine. The histidine residue of the catalytic triad was also substituted by either a serine or a tyrosine in some instances. The proenzyme cleavage site sequence was also variable, with APEA being the predominant N-terminal sequence. RT-PCR analyses indicated that CP1, CP2 and CP3 mRNA are all present in the bloodstream forms of T. congolense, showing that these variants are likely to be expressed. The conclusion of this study is that, given the high overall conservation of congopain genes in the genome, for the purpose of anti-disease vaccine, it is likely that a single immunogen will suffice to raise antibody able to inhibit all circulating congopain-like cysteine proteases. For chemotherapy however, a more in-depth enzymatic characterisation of the mutants, involving functional recombinant expression, will have to be undertaken.Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2008.
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Mucache, Hermogenes Neves. "Functional expression of Trypanosoma congolense pyroglutamyl peptidase type 1 and development of reverse genetics tools." Thesis, 2012. http://hdl.handle.net/10413/9915.
Full textAbstract:
Trypanosoma congolense is a protozoan parasite transmitted by tsetse flies. It causes bovine trypanosomosis, the major disease for livestock in sub-Saharan Africa. Control methods include trypanocidal drugs and vector control, but none is fully satisfactory, due to resistance and environmental issues. A method that would have the greatest impact on controlling the disease is vaccination. However, development of a conventional vaccine has been hampered by the mechanism of antigenic variation, which allows the parasite to evade the host’s immune system.
An alternative strategy in vaccine design is to target the bioactive compounds released by dead and dying trypanosomes. This approach is termed ‘‘anti-disease’’, and does not affect the survival of the parasite but targets the pathogenic factors released by the trypanosomes. The development of a successful anti-disease vaccine necessitates knowledge of all pathogenic factors involved in the disease process. Several macromolecules, primarily peptidases, have been implicated in the pathogenesis of trypanosomosis. Pyroglutamyl peptidase type I (PGP) was shown to be involved in abnormal degradation of thyrotropin- and gonadotropin-releasing hormones in rodents infected with T. brucei, but to date no data are available on the T. congolense PGP.
Molecular cloning and expression in E. coli of the coding sequence of T. congolense PGP, as well as the enzymatic characterisation of the recombinant protein, are reported here, completed by the development of reverse genetics tools for studies of gene function.
A 678 bp PCR fragment covering the complete open reading frame of PGP was cloned and sequenced. The deduced amino acid sequence showed 52% and 29% identity with the T. brucei and Leishmania major enzymes respectively. The catalytic residues Glu, Cys and His described in Bacilus amyloliquefaciens PGP are conserved in the T. congolense sequence. PGP was expressed in bacterial systems as a soluble active, 26 kDa enzyme. The recombinant enzyme showed activity specific for the fluorescent substrate pGlu-AMC, with a kcat/Km of 1.11 s-1μM. PGP showed activity in the pH 6.5-10 range, with maximal activity at pH 9.0. The enzyme was strongly inhibited by sulfhydryl-blocking reagents such as iodoacetic acid and iodoacetamide with a kass of 125 M-1 s-1 and 177 M-1 s-1 respectively. Antibodies raised in chickens against the recombinant enzyme allowed the detection of native PGP in both procyclic and bloodstream T.
congolense developmental stages, and displayed complete inhibition of the enzyme in vitro at physiological concentrations. To get insight into the role of PGP in parasite biology and trypanosomosis progression, two types of vectors for reverse genetics studies were developed. For RNA interference, a 400 bp 3′ end segment of the PGP open reading frame was cloned into the plasmid p2T7Ti, that will allow PGP gene down-regulation upon integration into the genome of an engineered tetracycline-inducible strain such as TRUM:29-13. For gene knock-out, several rounds of molecular engineering were carried-out in order to create two plasmid vectors, pGL1184-based (blasticidin resistance) and pGL1217-based (neomycin resistance), each bearing 200 bp-long regions at the 5′ and 3′ ends of the PGP open reading frame. In subsequent studies, taking advantage of the recent advances in culture and transformation of T. congolense, these plasmids will allow the creation of single and double knock-out mutants of PGP.Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.
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Vather, Perina. "Vivapain : a cysteine peptidase from Trypanosoma vivax." Thesis, 2010. http://hdl.handle.net/10413/4775.
Full textAbstract:
African animal trypanosomosis is a devastating disease affecting livestock mainly found in sub-Saharan Africa. This disease is known as nagana and is transmitted by the trypanosome parasite from the tsetse fly vector to a mammalian host. There are three African trypanosomes namely Trypanosoma vivax, T. congolense and T. brucei brucei that are the causative agents responsible for this disease in African cattle. This disease is serious since it not only affects livestock but also has a negative impact on the sub-Saharan African economy. There is, therefore, a great demand for better control methods of the disease and suitable diagnostic methods. Current control measures such as the use of trypanocidal drugs, tsetse fly eradication methods and trypanotolerant cattle have become inadequate. The defence mechanism of the trypanosome to continuously change its surface coat by a process of antigenic variation has made it impossible to produce a suitable vaccine. Therefore, chemotherapy is still one of the key approaches for control of this wasting disease. The long existence of the current trypanocidal drugs has allowed the development of drug resistance. The development of new chemotherapeutic drugs is focused on targeting the pathogenic factors such as parasite cysteine peptidases that contribute to the disease. Vivapain is the main cysteine peptidase of T. vivax and shares high sequence identity with congopain, the main cysteine peptidase of T. congolense, which was previously shown to be a pathogenic factor contributing to trypanosomosis. Vivapain, thus, has potential as a target for chemotherapeutic drug design. Hence, the first part of this study involved the recombinant expression and enzymatic characterisation of vivapain for future production of new synthetic inhibitors for the use in new trypanocidal drugs. The catalytic domain of vivapain (Vp) was recombinantly expressed in the Pichia pastoris yeast expression system and enzymatically characterised. The main finding from this study was that Vp was only able to hydrolyse a substrate if the P2 position was occupied by either a hydrophobic Phe or Leu residue. Vp was also found to be active close to physiological pH and was inhibited by the reversible cysteine peptidases, leupeptin, antipain and chymostatin and the irreversible cysteine peptidases L-trans-epoxysuccinyl-leucylamido (4-guanidino) butane (E-64), iodoacetic acid (IAA) and iodoacetamide (IAN). A further important aspect of controlling trypanosomosis is the diagnosis of the disease. Clinical, parasitological, molecular and serological techniques have been applied and used to diagnose trypanosomosis. One of the most promising serological techniques has proven to be the enzyme-linked immunosorbent assay (ELISA), more specifically the antibody and antigen detection ELISAs. The main requirement for this technique is a readily available and reproducible antigen such as that produced by recombinant expression. While there are recombinant antigens that are available to be used to detect T. congolense, T. brucei brucei and even T. evansi infections, there are none available to detect T. vivax infections. In the second part of this study, a mutant inactive full length form of vivapain (FLVp) was expressed in a bacterial expression system for the detection of T. vivax infections. Antibodies against this antigen were produced in both chickens and mice. Both the chicken IgY and mice sera were able to detect the recombinant FLVp in western blots. The mice sera were also able to detect native vivapain in a T. vivax lysate, which is very promising for future use of the FLVp antigen and the corresponding antibodies in diagnosis of T. vivax infections in sera of infected animals.Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2010.
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Kangethe, Richard Thiga. "Gene disruption of TcoCATL (Congopain) and oligopeptidase B, pathogenic factors of African trypanosomes." Thesis, 2011. http://hdl.handle.net/10413/8708.
Full textAbstract:
African trypanosomosis is a parasitic disease in man and animals caused by protozoan parasites of the genus Trypanosoma. T. congolense, T. vivax and T. brucei brucei cause nagana in cattle. The variable nature of the parasite surface coat has hindered the development of an effective vaccine. An option for developing vaccines and chemotherapeutic agents against trypanosomosis is to target pathogenic factors released by the parasite during infection, namely an “anti-disease” approach. Two pathogenic factors released during infection are oligopeptidase B (OPB) and TcoCATL (congopain). TcoCATL, a major lysosomal cysteine peptidase, is a member of the papain family C1 cysteine peptidases. RNA interference (RNAi) was used to down-regulate the expression of TcoCATL in T. congolense IL3000 TRUM183:29-13 parasites in vivo during mouse infections. TcoCATL RNAi was monitored in infected mouse blood by comparing the hydrolysis of Z-Phe-Arg-AMC and parasitaemia between mice in which RNAi was induced and control mice. Mice infected with parasites induced for TcoCATL RNAi had lower parasitaemia when compared to control mice. An attempt was also made at deleting the entire CATL gene array in both T. congolense IL3000 and T. brucei 427 Lister strains. The second pathogenic factor studied, OPB, is a cytosolic trypanosomal peptidase that hydrolyses peptides smaller than 30 amino acid residues, C-terminal to basic residues. In order to evaluate the role that OPB play during disease, RNAi was also applied to knock-down the expression levels of OPB in T. brucei T7T and T. congolense IL3000 TRUM183:29-13 strains (TbOPB and TcoOPB respectively). Oligopeptidase B null mutant strains (Δopb) were also generated in T. brucei brucei Lister 427. An attempt was also made to generate OPB null mutants in T. congolense IL3000 parasites. Western blot analysis of the knock-down experiments using chicken anti-TcoOPB peptide IgY showed that only TbOPB levels were reduced in T. brucei T7T parasites induced for RNAi when compared to TcoOPB RNAi induced cultures. Quantitative assessment of a fourteen day induction experiment for OPB RNAi in T. brucei showed an 87% reduction in TbOPB levels when compared to levels on day one. There was no growth effect observed in T. brucei parasites cultured in vitro and induced for TbOPB RNAi. It was concluded that TbOPB is not necessary for the in vitro survival of T. brucei parasites, thus making the generation of OPB null mutants possible. Δopb T. brucei parasites were successfully generated and grew normally in vitro and were as virulent as wild type strains during infection in mice. Immunohistopatholgy of infected mouse testes revealed Δopb parasites in extra vascular regions showing that T. brucei OPB (TbOPB) is not involved in assisting T. brucei parasites to cross microvascular endothelial cells. Gelatin gel analysis of Δopb null mutants and wild type strains showed an increase in cysteine peptidase activity. Enzymatic activity assays were carried out to identify how closely related oligopeptidases are affected by knocking out TbOPB, and a significant increase of T. brucei prolyl oligopeptidase (TbPOP) activity was observed. However, western blot analysis did not show any increase of TbPOP protein levels in Δopb parasites, suggesting that either TbOPB is responsible for generating an endogenous inhibitor for TbPOP or that another POP-like enzyme might compensate for a loss in OPB activity in Δopb null mutants. This study made a significant contribution to an understanding of the interplay between different trypanosomal peptidases that are important pathogenic factors in trypanosomosis. It highlights the need to simultaneously target several trypanosomal peptidases to develop an effective vaccine or chemotherapeutic agents for African animal trypanosomosis.Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2011.
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Takada, Ayato. "Experimental Study on Mucosal Vaccination of Animals against Viral Infections." Doctoral thesis, 1996. http://hdl.handle.net/2115/32735.
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Jackson, Laurelle. "Enzymatic and crystallisation studies of CATL-like trypanosomal cysteine peptidases." Thesis, 2011. http://hdl.handle.net/10413/8688.
Full textAbstract:
African animal trypanosomosis or nagana is a disease in livestock caused by various
species of protozoan parasites belonging to the genus Trypanosoma particularly T.
congolense, T. vivax and T. b. brucei. Nagana is the most important constraint to livestock
and mixed crop-livestock farming in tropical Africa. Trypanosomes undergo part of their
developmental life in their insect vector, the tsetse fly and part in their mammalian host.
Measures for eradicating the continent of the tsetse fly vector include insecticidal spraying,
targeting and trapping. Vaccine development has been hampered by the generation of an
inexhaustible collection of variant surface glycoproteins that trypanosomes possess and
allow for evasion of the host immune system. Anti-disease vaccines aimed at reducing the
symptoms of the disease rather than killing the parasite itself have been demonstrated as an
alternative approach. Trypanotolerant cattle are able to protect themselves from the
disease-associated symptoms. They are able to mount a better antibody response to the
CATL-like cysteine peptidase, TcoCATL, compared to trypanosusceptible breeds. Bovine
trypanosomosis, however, continues to be controlled primarily by trypanocidal compounds
such as isometamidium chloride, homidium and diaminazene that have been developed
more than 50 years ago and consequently drug resistance is widespread. Trypanosomal
cysteine peptidases have also been proven to be effective targets for chemotherapeutics.
TcrCATL, inhibited by the vinyl sulfone pseudopeptide inhibitor K11777, was effective in
curing or alleviating T. cruzi infection in preclinical proof-of-concept studies and has now
entered formal preclinical drug development investigation.
Understanding enzymatic as well as structural characteristics of pathogenic peptidases is
the first step towards successful control of the disease. To date no such characterisation of
the major cysteine peptidases from T. vivax has been conducted. Although the major
cysteine peptidase from T. vivax, TviCATL, has not been proven as a pathogenic factor yet,
its high sequence identity with the pathogenic counterparts such as TcrCATL and
TcoCATL hold much speculation for TviCATLs role in pathogenocity.
In the present study, native TviCATL was isolated from T. vivax Y486, purified and
characterised. TviCATL showed to have a general sensitivity to E-64 and cystatin and has a
substrate specificity defined by the S2 pocket. TviCATL exhibited no activity towards the
CATB-like substrate, Z-Arg-Arg-AMC but was able to hydrolyse Z-Phe-Arg-AMC, the
CATL-like substrate. Leu was preferred in the P2 position and basic and non-bulky
hydrophobic residues were accepted in the P1 and P3 positions respectively. Similar
findings were reported for TcoCATL. The substrate specificity of TviCATL and TcoCATL
does argue for a more restricted specificity compared to TcrCATL. This was based on the
Glu333 in TcrCATL substituted with Leu333 in TviCATL and TcoCATL. In the case of
TcrCATL, the Glu333 allows for the accommodation of Arg in the P2 position. Like other
trypanosomal cysteine peptidases, TviCATL was inhibited by both chloromethyl ketones,
Z-Gly-Leu-Phe-CMK and H-D-Val-Phe-Lys-CMK. Determining further structural and
functional characteristics as well as whether TviCATL, like the T. congolense homolog,
TcoCATL, acts as a pathogenic factor, would be important information to the designing of
specific chemotherapeutic agents.
To date, TcrCATL and TbrCATL (from T. b. rhodesiense) are the only trypanosomal
CATL-like cysteine peptidases been crystallised and their tructures solved. This advantage
has allowed for the directed design of synthetic peptidase inhibitors. The crystal structure
of TcoCATL will be of major significance to the design of specific chemotherapeutic
agents. Furtherrmore, understanding the dimeric conformation of TcoCATL is important
for vaccine design as immune responses are likely to recognise the dimer specific epitopes.
In the current study, the catalytic domain of TcoCATL and TviCATL, were recombinantly
expressed in Pichia pastoris and purified to homogeneity. The T. congolense cysteine
peptidase pyroglutamyl peptidase (PGP), also proven to be pathogenic in T. b. brucei, was
recombinantly expressed in E. coli BL21 (DE3) cells and also purified to homogeneity.
Purified cysteine peptidases along with previously purified TcoCATL dimerisation
mutants, TcoCATL (H43W) and TcoCATL (K39F; E44P), possessing mutated residues
involved in TcoCATL dimerisation, as well as the mutant proenzyme TcoCATL (C25A),
were screened for crystallisation conditions using the Rigaku robotic crystallisation suite.
One-dimensional needle-like crystals were found for TcoCATL (K39F; E44P).
Optimisation of the TcoCATL (K39F; E44P) crystals were analysed for X-ray diffraction.
The poor diffraction pattern prompted further optimisations for better crystal quality,
which is presently underway. The crystal structure of TcoCATL, with some of the residues
involved in dimerisation mutated, will be pivotal in understanding the dimerisation model.
Furthermore, the information about the structure will be valuable for vaccine design and
chemotherapeutics development.Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2011.
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Chang, Yuan-Chun, and 張淵俊. "The Development of Vaccination and the Application of Laboratory Animals in Vaccines." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/15353056579819573721.
Full textAbstract:
碩士大葉大學
生物產業科技學系碩士在職專班
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The development of vaccines has been the most important achievement in preventive medicine. In the past years epidemic diseases were incurable to human beings. However, since the development of vaccines, many infectious diseases are gradually controlled, and even the notorious disease smallpox has been disappeared in the world. Thus, vaccine has plays a key role and making a major contribution to the public health. The main types of vaccines can be divided into four: Inactivated vaccine, Attenuated vaccine, Subunit vaccine and DNA vaccine. The first two types are the traditional vaccines, although the system is old and simple, but is still the most widespread and effective biologics. As biotechnology and pharmaceutical industries have become more advanced in vaccine and medicine making, Subunit and DNA vaccines could be gain our attention in the future in terms of research and development trends. Laboratory animals have been in the use of scientific research for a long history, and the vaccine testing on Laboratory animals is very common. In fact, vaccine safety is extensively evaluated through animal safety studies before using by humans. The animal safety studies included in vitro and in vivo. Thus, scientists have performed many research in vitro and in vivo in the laboratory to prove the safety and effectiveness of medical products. For example, a new development of vaccine also must have considered numbers of animal testing data to support its safety effectiveness. As for the vaccine manufacturing process, both traditional and modern vaccines are prepared by using of laboratory animals to carry out. In terms of quality control, the results of animal testing can guarantee a product quality. For the future development of a new vaccine, still must rely on laboratory animals for vaccine research and study. Therefore, laboratory animals testing has plays a key role in vaccine development, and further to the public health.
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Green, Karin Kappmeier. "Strategy for monitoring and sustainable integrated control or eradication of Glossina brevipalpis and G.austeni (Diptera: Glossinidae) in South Africa." Thesis, 2002. http://hdl.handle.net/2263/29893.
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Compton, Justin A. "Ecology of common raccoon (Procyon lotor) in western Pennsylvania as related to an oral rabies vaccination program." 2007. http://www.etda.libraries.psu.edu/theses/approved/WorldWideIndex/ETD-1824/index.html.
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Crow, Linnet Jean Isobel. "Testing for passive transfer of immunity in foals, and an evaluation of the African horse sickness vaccination schedule." Thesis, 2005. http://hdl.handle.net/10413/4089.
Full textAbstract:
This thesis comprises an introductory review of the literature, followed by reports of two experiments which are presented in the form of scientific papers. For this reason, there may be some repetition between chapters, particularly in terms of experimental procedure. To avoid unnecessary repetition, a single list of references is given at the end of the thesis. For the sake of completeness , several appendices are attached to Chapters Two and Three which would not ordinarily be included in a scientific paper. The literature review looks at the passive transfer of immunity from the mare to the foal, the consequences of failure of passive transfer of immunity and different methods of testing whether the transfer of passive immunity has occurred. The review concludes with a discussion of vaccination programmes against African horse sickness. Trial One evaluated different tests for determining whether the transfer of passive immunity from mare to foal has occurred in order to determine which of these tests should be used preferentially. A single radial immunodiffusion test was used as the reference standard. A series of samples .was taken from a group of foals and tested using four methods: single radial immunodiffusion , glutaraldehyde coagulation, zinc sulphate turbidity and protein refractometer tests. Trial Two explored African horse sickness vaccination programmes, focusing on when to vaccinate foals for the first time. A series of samples was taken from a group of foals from birth until two months after their second set of African horse sickness vaccinations (one year old). The samples were tested for the presence of African horse sickness antibodies for each of the nine serotypes to determine when maternal immunity fades and to evaluate the effect of each vaccination on the level of immunity.Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.
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De, Castro Minique Hilda. "Sialotranscriptomics of the brown ear ticks, Rhipicephalus appendiculatus Neumann, 1901 and R. Zambeziensis Walker, Norval and Corwin, 1981, vectors of Corridor disease." Thesis, 2017. http://hdl.handle.net/10500/24735.
Full textAbstract:
Text in EnglishCorridor disease is an economically important tick-borne disease of cattle in southern Africa. The disease is caused by Theileria parva and transmitted by the vectors, Rhipicephalus appendiculatus and R. zambeziensis. There is currently no vaccine to protect cattle against T. parva that is permitted in South Africa. To develop recombinant anti-tick vaccines against Corridor disease, comprehensive databases of genes expressed in the tick’s salivary glands are required. Therefore, in Chapters 2 and 3, mRNA from the salivary glands of R. appendiculatus and R. zambeziensis was sequenced and assembled using next generation sequencing technologies. Respectively, 12 761 and 13 584 non-redundant protein sequences were predicted from the sialotranscriptomes of R. appendiculatus and R. zambeziensis and uploaded to public sequence domains. This greatly expanded the number of sequences available for the two vectors, which will be invaluable resources for the selection of vaccine candidates in future. Further, in Chapter 3, differential gene expression analysis in R. zambeziensis revealed dynamic expression of secretory protein transcripts during feeding, suggestive of stringent transcriptional regulation of these proteins. Knowledge of these intricate expression profiles will further assist vaccine development in future. In Chapter 4, comparative sialotranscriptomic analyses were performed between R. appendiculatus and R. zambeziensis. The ticks have previously shown varying vector competence for T. parva and this chapter presents the search for correlates of this variance. Phylogenetic analyses were performed using these and other publically available tick transcriptomes, which indicated that R. appendiculatus and R. zambeziensis are closely related but distinct species. However, significant expression differences were observed between the two ticks, specifically of genes involved in tick immunity or pathogen transmission, signifying potential bioinformatic signatures of vector competence. Furthermore, nearly four thousand putative long non-coding RNAs (lncRNAs) were predicted in each of the two ticks. A large number of these showed differential expression and suggested a potential transcriptional regulatory function of lncRNA in tick blood feeding. LncRNAs are completely unexplored in ticks. Finally, in Chapter 5, concluding remarks are given on the potential impact the R. appendiculatus and R. zambeziensis sialotranscriptomes may have on future vaccine developments and some future research endeavours are discussed.
Life and Consumer Sciences
Ph. D. (Life Sciences)