Dissertations / Theses on the topic 'Trypanosomiasis in cattle – Zambia'

Made available in DSpace on 2014-06-11T19:31:10Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-23Bitstream added on 2014-06-13T18:41:54Z : No. of bitstreams: 1 teixeira_mca_dr_jabo.pdf: 2885202 bytes, checksum: 80086a83bb46ca2cb05bc1ae5ea75a8d (MD5)<br>Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)<br>Trypanosoma evansi é patogênico para a maioria dos animais, acometendo bovinos, bubalinos, caprinos, ovinos, suínos, cães, quatis, capivaras, camelos e outras espécies animais em áreas tropicais e subtropicais do globo terrestre sendo, no Brasil, a doença endêmica no pantanal mato-grossense. O presente estudo teve como fito principal estudar a evolução clínica, as alterações hematológicas, bioquímicas sérica, imunológicas e anatomopatológicas de bovinos infectados experimentalmente com T. evansi. Para tal, foram utilizados oito bovinos, clinicamente sadios e sorologicamente negativos para T. evansi. Três foram mantidos como testemunhos e cinco inoculados com T. evansi. Exames físicos, parasitológicos, hematimétricos e bioquímicos séricos (proteínograma, índice ictérico e glicose) e do líquido cefalorraquidiano foram realizados. Nos exames físicos realizados nos bovinos até 525° DAI não foi notada nenhuma anormalidade clínica com relação à temperatura retal, batimentos cardíacos, frequência respiratória, movimentos ruminais, aspectos de membranas mucosas (nasal, conjuntival, oral, vaginal e/ou prepucial) e dos linfonodos externos (mandibulares, maxilares, parotídeos, cervicais superficiais, sublíacos e mamários). A presença de tripomastigotas foi demonstrada através da prova biológica nos bovinos 01, 06 e 08 no15° DAI, bovinos 06 e 07 no 30° DAI, bovinos 01 e 06 no 45° DAI, bovino 06 no 60° DAI, bovino 01 no 75° DAI. As contagens de hemácias, os teores de hemoglobina e os volumes globulares dos bovinos, experimentalmente infectados, variaram dentro dos limites de normalidade para a espécie bovina. O VGM, HGM e CHGM, apresentam alterações pontuais.<br>Trypanosoma evansi are pathogenic to most of animals, affecting cattle, buffaloes, goats, sheep, pigs, dogs, coatis, capybaras, camels and other animals in tropical and subtropical areas of the globe, and, in Brazil, it causes an endemic disease in the Pantanal Mato Grosso. This study primarily aimed to study the clinical, hematological, biochemical, immunological and pathological alterations in cattle experimentally infected with T. evansi. For this purpose, we used eight animals, clinically healthy and serologically negative for T. evansi. Three animas were kept as evidence and five were inoculated with T. evansi. Physical, parasitological, hematological and serum biochemical (proteins, icteric index and glucose) and cerebrospinal fluid examination were performed. In the physical examination conducted in cattle up to 525th DAI were not observated any clinical abnormality in concerning rectal temperature, heart rate, respiratory rate, ruminal movements, aspects of the mucous membranes (nasal, conjunctival, oral, vaginal and / or specimen) and external nodes (mandibular, maxillary, parotid, superficial cervical, breast and sublíacos). The presence of trypomastigotes was demonstrated by bioassay in cattle 01, 06 and 08 no 15th DAI, cattle 06 and 07 at 30° DAI, cattle 01 and 06 on the 45 th DAI, cattle 06 in 60 th DAI, cattle 01 in 75 th DAI. Red blood cells counts, hemoglobin content and volume cell of experimentally infected cattle were within normal limits for the bovine species. The MCV, MHC and MCHC, showed specific changes. Physical examination of the cerebrospinal fluid did not show alterations in appearance and coloration. Morever, using the Giensa-stained blood smears, buffy coat technique (BCT) and mouse inoculation procedure were negative for T. evansi tripomastigote. Serum protein concentrations, identified 26 proteins with molecular weights ranging from 20 to 245 KD.

Over the past two decades movement of cattle towards the north of Uganda has enabled the Trypanosoma brucei rhodesiense focus in south-eastern Uganda to spread into previously unaffected districts. This thesis brings together important epidemiological data regarding the impact of mass cattle drug treatment on the point prevalence of several different species of trypanosome in a newly endemic area of human sleeping sickness. Crucially the findings illustrate mass drug treatment is effective in reducing the prevalence of T. b. rhodesiense in cattle, thus minimising the reservoir potential of these animals in the epidemiology of human disease. During 2006 a control programme was launched to halt the northward spread of this zoonotic parasite. This programme, entitled ‘Stamping Out Sleeping Sickness’ (SOS) proposed to reduce the prevalence of Human African Trypanosomiasis (HAT) in the newly affected districts by reducing the prevalence of this parasite in the main animal reservoir of infection – domestic cattle. Cattle were mass treated using trypanocides to clear infections. Previous work demonstrated the prevalence of T. brucei s. l. and T. b. rhodesiense in cattle was higher in the districts of Dokolo and Kaberamaido than in the other SOS intervention districts (Selby 2011). To determine whether animals in these areas were also exposed to pathogenic cattle trypanosomes samples were screened for the presence of T. vivax and T. congolense savannah using PCR. Chapter three of this thesis determined the prevalence of these trypanosomes in cattle in these districts. Before treatment had taken place the prevalence of T. vivax was 2% (4/200, 95% CI 3.57 – 0.12%) in Dokolo and 7.3% (21/310, 95% CI 10.17 - 4.24 %) in Kaberamaido. The prevalence of T. congolense savannah at baseline was 3.5% (7/200, 95% CI 7.08–1.42 %) in Dokolo and 9.1% (21/230, 95% CI13.6–5.7 %) in Kaberamaido. Monitoring was conducted three, nine and 18 months post treatment and both pathogens were detected at all time points. The impact the treatment had on point prevalence varied by trypanosome species and between the two districts. Several clusters of villages in Dokolo and Kaberamaido continued to report cases of HAT after the initial SOS intervention due in part to their proximity to livestock markets (Batchelor et al., 2009). In 2008 re-treatment of these ‘high risk’ areas was undertaken. Monitoring was performed before and six months after treatment. Cattle blood samples were collected at 20 village sites from ten ‘case-positive villages’ (from which human sleeping sickness cases had been reported six months prior to June 2007) and from ten ‘case-negative villages’ (no reported human sleeping sickness cases six months prior to June 2007). These samples were screened for all of the aforementioned trypanosomes using species specific PCR protocols. Chapter five details the results of this screening, and assessed whether re-treatment in Dokolo and Kaberamaido was effective in reducing the prevalence of trypanosomiasis. The re-treatment had a dramatic effect, significantly reducing the point prevalence of overall trypanosomiasis in the 20 villages screened from 38.1% (95% CI = 40.5 – 35.79%) at baseline to 26.9% (95% CI 28.96 – 24.97, p < 0.0001) at six months. Looking at each species separately, point prevalence of three out of four detected species of trypanosome fell significantly, including T. b. rhodesiense, which was reduced to 25% of its baseline prevalence. Finally the two SOS treatment cycles were compared both statistically and spatially with emphasis on trends at village level and the occurrence of mixed infections.

Tsetse flies, Glossina species, are the biological vectors of Trypanosoma species which cause animal African trypanosomiases (AAT) in livestock (especially cattle) in sub-Saharan Africa. This disease is often fatal without treatment and negatively impacts on rural, agricultural and economic development. On the Jos Plateau, north central Nigeria, AAT was historically of little significance due to the presumed absence of tsetse and Fulani pastoralists were encouraged to settle there. But over the last 30 years, the disease has become widespread and highly prevalent in the area. This has been attributed to the expansion of tsetse on the plateau, frequent migrations of cattle to areas with higher tsetse densities and the presence of other biting flies which serve as mechanical vectors. In the current study, the presence and abundance of tsetse was determined in selected villages using biconical tsetse trap surveys. The low number of flies trapped suggests that tsetse expansion has been very limited within the plateau but the fact that trypanosome DNA was present in over half of these flies implicates them in AAT transmission. The migration of a herd of cattle was also tracked and during the period, blood samples were collected from the cattle and examined for trypanosomes using molecular techniques. Despite prophylactic treatment and deltamethrin sprays, results showed that a significant proportion of the animals (52%) had become infected with T. vivax over the migration period. Tsetse flies (G. palpalis) were also slightly more abundant in some of parts of the migration area. Potential mechanical vectors (Stomoxys spp. and Tabanidae) were trapped and results obtained from the examination of their mouthparts for trypanosomes indicate their involvement in transmission. However, it is difficult to make any definite conclusions about their overall contribution which is thought to be minimal and more studies are needed to clarify their significance. It is concluded that trypanosomiasis risk from tsetse on the Jos Plateau is currently low and seasonal migration appears to be the main driver of AAT transmission by exposing cattle to more tsetse for longer periods. Other biting flies may play a limited role which remains undetermined. Continued monitoring of cattle and tsetse across the plateau over the next few years is important and the careful use of trypanocides and insecticide treated cattle is recommended as an appropriate control strategy.

Thesis (PhD)--Stellenbosch University, 2013.<br>ENGLISH ABSTRACT: In Uganda, cattle are an important reservoir of Trypanosoma brucei rhodesiense, a parasite that causes human African trypanosomiasis or sleeping sickness. We developed mathematical models to examine the transmission of T. b. rhodesiense by tsetse vector species, Glossina fuscipes fuscipes in a host population that consists of humans, domestic and wild mammals, and reptiles. The models were developed and analysed based on the situation in Tororo district in Southeastern Uganda, where sleeping sickness is endemic and which has a cattle and human population of 40, 000 and 500, 000, respectively. Assuming populations of cattle and humans only, the impact of mass chemoprophylaxis and vector control through insecticide-treated cattle (ITC) is evaluated. Keeping 12% or 82% of the cattle population on insecticides that have an insecticidal killing effect of 100% at all times or trypanocides that have 100% efficacy, respectively, can lead to the control of T. b. rhodesiense in both humans and cattle. Optimal control of T. b. rhodesiense is shown to be achieved through ITC alone or a combination of chemoprophylaxis and ITC, the former being the cheapest control strategy. Allowing for the waning effect of insecticides and including wildhosts, T. b. rhodesiense control can be achieved by keeping 21% or 27% of the cattle population on insecticides through whole-body or restricted application, respectively. Restricting the treatment of insecticides to adult cattle only would require 24% or 33% of the adult cattle population to be kept on insecticides through whole-body or restricted application, respectively, to control T. b. rhodesiense. A cost-effectiveness and benefit-cost analysis of using ITC to control T. b. rhodesiense show that restricted application of insecticides is a cheaper and more beneficial strategy compared to whole-body treatment. The results of the study show that the restricted application of insecticides on cattle provides a cheap, safe and farmer-based strategy for controlling tsetse and trypanosomiasis.<br>AFRIKAANSE OPSOMMING: In Uganda is beeste ’n belangrike reservoir van Trypanosoma brucei rhodesiense, ’n parasiet wat tripanosomiase of slaapsiekte in mense veroorsaak. Ons het wiskundige modelle ontwikkel wat die oordrag van T. b. Rhodesiense deur tesetse vektor spesies, Glossina fuscipes fuscipes in ’n draer populasie wat bestaan uit mense, mak en wilde diere en reptiele, ondersoek. Die modelle was ontwikkel en geanaliseer gebaseer op die oordrag situasie in die Tororo distrik in Suidoostelike Uganda, ’n gebied waar slaapsiekte endemies is en wat ’n populasie van 40, 000 beeste en 500, 000 mense het. Die impak van massa chemoprofilakse en vektor beheer deur insekdoder-behandelde beeste is gevalueer onder die aanname van bees en mens populasies alleenlik. Beheer oor T. b. Rhodesiense in beide mense en beeste kan verkry word deur of 12% van die bees populasie te behandel met ’n insekdoder wat 100% effektief is ten alle tye of 82% van die bees populasie te behandel met tripanosiedes wat 100% effektief is. Daar is aangetoon dat optimale beheer van T. b. Rhodesiense bereik kan word deur die gebruik van insekdoders alleenlik of ’n kombinasie van insekdoders en chemoprofilakse, hoewel eersgenoemde die goedkoopste strategie is. Wanneer die kwynende effek van insekdoders asook wilde diere as draers in ag geneem word, kan T. b. Rhodesiense beheer verkry word deur 21% van beeste se hele liggaam met insekdoders te behandel of 27% gedeeltelik te behandel. As slegs volwasse beeste met insekdoders behandel word, moet 24% se hele liggaam of 33% gedeeltelik behandel word vir beheer van T. b. Rhodesiense. ’n Koste-effektiwiteit en voordeel-koste analise van insekdoders as beheermaatstaf vir T. b. Rhodesiense toon aan dat gedeeltelike behandeling van die bees se liggaam die goedkoper en meer voordelige strategie is in vergelyking met behandeling van die hele liggaam. Die resultate van die studie wys dat gedeeltelike behandeling van beeste met insekdoders ’n goedkoop, veilige en landbouer-gebaseerde strategie is om tsetse en tripanosomiase te beheer.

The most important tick-borne diseases (TBDs) occurring in Zambia that affect domestic animals, particularly cattle and small ruminants, are theileriosis (East Coast fever and Corridor disease), anaplasmosis (gall sickness), babesiosis (red water), and heartwater (cowdriosis). Of these, theileriosis is the most important, causing significantly more deaths than the other tick-borne diseases combined. Despite their importance, little is known about the occurrence and prevalence of haemoparasites in cattle in the communal areas of Zambia. Clinical signs and post mortem lesions are pathognomonic of mixed tick-borne infections especially babesiosis, anaplasmosis and East Coast fever (ECF). The main objective of this study was, therefore, to screen selected communal herds of cattle for tick-borne haemoparasites and identify the tick vectors associated with the high cattle mortalities due to suspected TBDs in the local breeds of cattle grazing along the banks of the Chambeshi River in Mungwi, Zambia. East Coast fever is endemic to the district of Mungwi, Northern Province, Zambia and vector control using acaricides has proved to be very costly for the small scale farmers. Also, Mungwi experiences increased cattle mortalities between December to March and May to July. All age groups of cattle are affected. A total of 299 cattle blood samples were collected from July to September 2010 from Kapamba (n=50), Chifulo (n=102), Chisanga (n=38), Kowa (n=95) and Mungwi central (n=14) in the Mungwi District, Northern Province, Zambia. Ticks were also collected from the sampled cattle from April to July 2011. DNA was extracted and the parasite hypervariable region of the small subunit rRNA gene was amplified and subjected to the reverse line blot (RLB) hybridization assay. The results of the RLB assay revealed the presence of tick-borne haemoparasites in 259 samples occurring either as single or mixed infections. The most prevalent species present were the benign Theileria mutans (54.5%) and T. velifera (51.5%). Anaplasma marginale (25.7%), Babesia bovis (7.7%) and B. bigemina (3.3%) were also detected in the samples. Nine percent of the samples tested negative for the presence of haemoparasites. In a number of samples (4%) the PCR products failed to hybridize with any species-specific probes but hybridized only with the genus-specific probes which could suggest the presence of a novel species or variant of a species. Of the four Theileria species known to occur in Zambia (T. parva, T. mutans, T. velifera and T. taurotragi), T. parva is the most economically important, causing Corridor disease in the Southern, Central, Lusaka and the Copper-belt provinces, while causing ECF in the Northern and Eastern provinces of Zambia. In our study, only one sample (from Kapamba) tested positive for the presence of T. parva. This was an unexpected finding; also because the tick vector, Rhipicephalus appendiculatus, was identified on animals from Kowa (14%), Chisanga (8.5%), Chifulo (6%) and Kapamba (1.4%). We can only speculate that the RLB hybridization assay may not have been able to detect the parasite in the animals sampled due to a too low parasitaemia. The samples should also be subjected to the T. parva specific real-time PCR assay to determine a more accurate T. parva prevalence in cattle in the Mungwi district, Northern Province. In Zambia, Babesia bovis and B. bigemina are recognized as being of economic importance in cattle. In our study, B. bovis was present in 7.7% of the sampled animals and B. bigemina in 3.3% of the animals. We detected B. bovis in all of the five sampled areas with the highest detection in Mungwi central (14.3%) and Kowa (10.5%). As expected, the tick vector Rhipicephalus (Boophilus) microplus was identified from animals from all of these areas. Babesia bigemina was only reported from Kowa (10.5%). The most abundant ticks identified from the sampled animals from Kowa were Rhipicephalus (Boophilus) decoloratus (36.3%) and Rhipicephalus (Boophilus) microplus (8.8%). These tick vectors have been implicated in the transmission of B. bigemina. Our findings are in concordance with results obtained by other authors who have speculated that an increase in the detection of B. bovis may indicate that B. bovis is becoming endemic in this part of the country. This could be due to uncontrolled movement of cattle that frequently occurs within Zambia. Heartwater (cowdriosis) is caused by Ehrlichia ruminantium, a rickettsial disease that affects domestic and wild ruminants. In Zambia, heartwater is mainly a disease of cattle, although outbreaks in sheep and goats have been reported and recorded. In our study, only one sample (from Kapamba) tested positive for the presence of E. ruminantium even though Amblyomma variegatum ticks were identified from 52.9% of the sampled animals from all study areas. The cattle sampled in our study are not regularly dipped and no game has been spotted in cattle grazing areas. It is possible that these cattle may have attained a state of endemic stability to heartwater. It is also possible that the RLB hybridization assay may not have been sensitive enough to detect E. ruminantium infections if the parasitaemia was very low. Samples should also be subjected to the E. ruminantium-specific pCS20 real-time PCR assay to determine more accurately the E. ruminantium prevalence in cattle in the Mungwi district, Northern Province. Anaplasma marginale (the causative agent of bovine anaplasmosis) has previously been shown to be present in all the provinces of Zambia and is the only Anaplasma species of importance to cattle in Zambia. In our study, 25.7% of the sampled cattle tested positive for A. marginale; it was detected in all areas except Chisanga. Amblyomma variegatum was identified from 52.9% of the sampled cattle, and Rhipicephalus (Boophilus) microplus from 12.1% of the cattle. Rhipicephalus (Boophilus) microplus has been incriminated as being a vector of A. marginale. Furthermore, three samples (from Kowa) tested positive for the presence of Anaplasma centrale. To our knowledge, no vaccination regime using A. centrale is being conducted in the Mungwi district of Zambia. The presence of A. centrale is, therefore, an interesting finding. The results of our study suggest that the cause of cattle mortalities in Mungwi during the winter outbreaks is mainly due to A. marginale, B. bovis and B. bigemina infections. This was confirmed by the results of the RLB hybridization assay, clinical manifestation of the disease in the affected cattle (own observation) and the tick species identified on the animals. It appears that in Mungwi, babesiosis due to B. bovis mostly infects cattle above one year of age. Calves appear to be less affected by B. bovis infection. There is need for further epidemiological surveys in Mungwi district, Northern Province, Zambia to get a better understanding of the epidemiology of these tick-borne haemoparasites affecting cattle. We conclude that integrated control policies should be developed to take account of multi-species pathogen communities that are commonly associated with clinical and sub-clinical TBD infections in Zambia.<br>Dissertation (MSc)--University of Pretoria, 2012.<br>Veterinary Tropical Diseases<br>unrestricted

Two-dimensional (2-D) motion sensors are activity motion sensors that use electronic accelerometers to record the lying, standing and walking behaviour of animals. They were used in this study with the aim of monitoring and quantifying the movement behaviour of traditionally managed cattle in the context of improving animal health and production in rural sub-Saharan Africa. Improvements in animal health and production could be made if data can be automatically collected on large numbers of animals and over prolonged periods of time. This data can then be used by stakeholders in making management and disease control decisions. This study was designed to assess whether the 2-D motion sensors were suitable for use on traditionally managed cattle in Kasero and Makale, two veterinary camps in Petauke District, Eastern Province of Zambia. It further aimed to provide a baseline for future research on traditional cattle movement behaviour. The study was carried out in a region where trypanosomiasis and tick-borne diseases are endemic and low haemoglobin values are often associated with these and other parasitic infections. An assessment was made on the effect on cattle movement behaviour of a treatment directed against tsetse-transmitted (trypanosoma congolense (Savannah type), trypanosoma vivax and trypanosoma brucei), tick-transmitted (theileria parva, anaplasma spp. and babesia spp.) and pasture-transmitted pathogens of African cattle. A structured questionnaire on livestock ownership and management practices showed that cattle owners considered trypanosomiasis and theileriosis the main constraints to improved cattle health and production in their traditional crop-livestock mixed farming system. A baseline study was conducted in which haemoglobin values were measured in 432 cattle in the two areas. In each area, ten pairs of co-grazing cattle were selected on the basis of one high and one low haemoglobin value in each pair. The co-grazing pairs were age and sex matched. Each animal had a motion sensor placed on its hind leg, to continuously measure and record its activity for two weeks. There were significant differences in haemoglobin levels between the two camps with Makale having lower levels than Kasero. Baseline data indicated that a larger proportion of sampled animals in Makale had trypanosomiasis while those in Kasero had theileriosis. Molecular parasitological results showed that the proportion of animals sampled in Makale that had trypanosomiasis was greater (21.4% [95%CI: 16.4 – 27.1]) than that in Kasero (1.4% [95%CI: 0.5 – 4.1]). However, Kasero had a greater proportion of animals positive for theileriosis (25.6% [95%CI: 20.2 – 31.9]) than Makale (2.4% [95%CI: 1.0 – 5.2]). A total of 204 cattle were screened for a three week treatment study in Makale. From this number, 40 animals with low circulating haemoglobin levels (<8g/dl) were paired and investigated for differences in movement behaviour patterns between treated and non- treated cattle. Analysing the sensor data using principal components analysis (PCA) revealed that the treated animals (which had higher mean haemoglobin values at the end of the study) were clustered more closely on the score plots than the control animals (which had lower mean haemoglobin values). The numbers of steps taken by high haemoglobin cattle in both studies were significantly higher than the low haemoglobin cattle. This, coupled with the PCA results suggests an association between cattle haemoglobin levels and their movement patterns.

Cattle ticks and tick-borne infections (TBIs) undermine cattle health and productivity in the whole of sub-Saharan Africa (SSA) including Nigeria. In this West African country, two thirds of the cattle population are reared in the central-northern regions, kept under the traditional pastoral husbandry of Fulani herders. Under the Fulanis’ management, cattle are grazed extensively, being exposed to infestation by several tick genera (i.e. Amblyomma, Hyalomma, and Rhipicephalus spp., sub-genus Boophilus spp. included), vectors of the causative agents of the most important bovine TBIs in West Africa: anaplasmosis, babesiosis and ehrlichiosis (cowdriosis). Nevertheless, the Fulani pastoralists do not usually employ chemicals to control ticks in their cattle, merely relying on traditional methods (i.e. manual removal of the most conspicuous specimens). This approach, however, does not prevent cattle from being re-infested, leaving the animals challenged by a broad variety of other tick species, most of which are vectors of economically relevant TBIs. Knowledge of tick and TBIs occurrence is an essential pre-requisite to assist field diagnosis and devising effective control strategies for a given area. Existing information on tick infestation of cattle in Nigeria is rather out-dated, mostly derived from studies carried out in the south of the country. Similarly, all studies published to date on cattle TBIs in the country do not include any molecular analysis, being based instead on cytological and/or serological diagnostics. Therefore, the aim of the present thesis was to assess the presence of cattle ticks and TBIs occurring in an area of Central Nigeria (i.e. Plateau State). This is a densely populated area with traditionally managed cattle, where no acarides have historically been employed on livestock. The work undertaken herein firstly reviews the information available to date on ticks and TBIs known to be endemic in Nigerian cattle, identifying gaps present in the existing knowledge, leading to the rationale of this study. An initial survey was conducted documenting the tick species infesting cattle in Central Nigeria, in order to assess the infestation rate of surveyed animals at the time of the year when the tick load on the host is known to be most abundant (i.e. the wet season). The survey provided novel information on tick populations in cattle in Nigeria disclosing the presence of a broad variety of species, most of which are vectors of hazardous TBIs. In order to conduct a molecular diagnosis of the TBIs within the study area, a novel methodology was developed (i.e. reverse line blotting, RLB). The application of this approach was based on a thorough review of its application to the diagnosis of TBIs worldwide as well as in SSA. The optimisation of the RLB at the University of Edinburgh to enable the detection of a broad-spectrum of TBIs in Nigeria, caused by an array of five genera of microorganisms (i.e. Ehrlichia and Anaplasma, Theileria and Babesia, Rickettsia spp.) is presented. The assessment of the analytical sensitivity of this technique for the detection of Anaplasma marginale, a highly endemic tick-borne pathogen in SSA, demonstrated a detection threshold of ≥ 7 infected cells (keeping the limit of a natural infection). The occurrence of TBIs in cattle in the study area was assessed during a large-scale epidemiological survey through the application of the validated RLB. This study disclosed the occurrence of a high prevalence of several bovine TBIs in Central Nigeria, some of which are of great veterinary and zoonotic concern. The RLB enabled the detection of carrier status as well as of numerous multiple infections (69.5%, 95% CI: 65.5–73.6%). Based on the findings presented, endemic stability for highly prevalent haemoparasites (i.e. Theileria mutans, Theileria velifera, Theileria taurotragi, Anaplasma marginale, Ehrlichia species Omatjenne) is postulated, whereas a more instable epidemiological scenario is hypothesized for other microorganisms (i.e. Anaplasma centrale and Babesia bovis), which might be connected with outbreaks of clinically apparent disease, sporadically seen in the study area. The effect of a monthly tsetse-borne trypanosomiasis-focused control programme (based on the application 0.005% deltamethrin spray formulation, applied only to the lower quarters of cattle) on the kinetics of bovine TBIs was assessed at the village level. Longitudinal monitoring of control and treated cattle was conducted over the period of eleven months. Results generated provide input to the improvement of future control strategies to be rolled out across SSA, aiming to achieve an integrated control of both trypanosomiasis and TBIs. The present thesis contributes to a better understanding of the epidemiology of bovine TBIs in Nigeria as well as in the rest of West Africa, using a highly sensitive tool of wide applicability. These findings will be shared with the local pastoralist communities to further promote effective yet sustainable, vector control, in tune with the traditional long-established practices.

Orientador: Luiz Carlos Marques<br>Banca: Rosangela Zacarias Machado<br>Banca: Fabiano Antonio Cadioli<br>Banca: Percílio Brasil dos Passos<br>Banca: Thais Helena Constantino Patelli<br>Resumo: Trypanosoma evansi é patogênico para a maioria dos animais, acometendo bovinos, bubalinos, caprinos, ovinos, suínos, cães, quatis, capivaras, camelos e outras espécies animais em áreas tropicais e subtropicais do globo terrestre sendo, no Brasil, a doença endêmica no pantanal mato-grossense. O presente estudo teve como fito principal estudar a evolução clínica, as alterações hematológicas, bioquímicas sérica, imunológicas e anatomopatológicas de bovinos infectados experimentalmente com T. evansi. Para tal, foram utilizados oito bovinos, clinicamente sadios e sorologicamente negativos para T. evansi. Três foram mantidos como testemunhos e cinco inoculados com T. evansi. Exames físicos, parasitológicos, hematimétricos e bioquímicos séricos (proteínograma, índice ictérico e glicose) e do líquido cefalorraquidiano foram realizados. Nos exames físicos realizados nos bovinos até 525° DAI não foi notada nenhuma anormalidade clínica com relação à temperatura retal, batimentos cardíacos, frequência respiratória, movimentos ruminais, aspectos de membranas mucosas (nasal, conjuntival, oral, vaginal e/ou prepucial) e dos linfonodos externos (mandibulares, maxilares, parotídeos, cervicais superficiais, sublíacos e mamários). A presença de tripomastigotas foi demonstrada através da prova biológica nos bovinos 01, 06 e 08 no15° DAI, bovinos 06 e 07 no 30° DAI, bovinos 01 e 06 no 45° DAI, bovino 06 no 60° DAI, bovino 01 no 75° DAI. As contagens de hemácias, os teores de hemoglobina e os volumes globulares dos bovinos, experimentalmente infectados, variaram dentro dos limites de normalidade para a espécie bovina. O VGM, HGM e CHGM, apresentam alterações pontuais.<br>Abstract: Trypanosoma evansi are pathogenic to most of animals, affecting cattle, buffaloes, goats, sheep, pigs, dogs, coatis, capybaras, camels and other animals in tropical and subtropical areas of the globe, and, in Brazil, it causes an endemic disease in the Pantanal Mato Grosso. This study primarily aimed to study the clinical, hematological, biochemical, immunological and pathological alterations in cattle experimentally infected with T. evansi. For this purpose, we used eight animals, clinically healthy and serologically negative for T. evansi. Three animas were kept as evidence and five were inoculated with T. evansi. Physical, parasitological, hematological and serum biochemical (proteins, icteric index and glucose) and cerebrospinal fluid examination were performed. In the physical examination conducted in cattle up to 525th DAI were not observated any clinical abnormality in concerning rectal temperature, heart rate, respiratory rate, ruminal movements, aspects of the mucous membranes (nasal, conjunctival, oral, vaginal and / or specimen) and external nodes (mandibular, maxillary, parotid, superficial cervical, breast and sublíacos). The presence of trypomastigotes was demonstrated by bioassay in cattle 01, 06 and 08 no 15th DAI, cattle 06 and 07 at 30° DAI, cattle 01 and 06 on the 45 th DAI, cattle 06 in 60 th DAI, cattle 01 in 75 th DAI. Red blood cells counts, hemoglobin content and volume cell of experimentally infected cattle were within normal limits for the bovine species. The MCV, MHC and MCHC, showed specific changes. Physical examination of the cerebrospinal fluid did not show alterations in appearance and coloration. Morever, using the Giensa-stained blood smears, buffy coat technique (BCT) and mouse inoculation procedure were negative for T. evansi tripomastigote. Serum protein concentrations, identified 26 proteins with molecular weights ranging from 20 to 245 KD.<br>Doutor

Factors affecting the epidemiology and impact of trypanosomiasis in livestock were studied in households in Mambwe District in an area extending from the Luangwa river valley to the edge of the eastern plateau. A structured questionnaire survey on demography, migration and farm activities of householders showed that about 84% of the households depended on farming for their income, mainly cotton growing. A cross-sectional study of 649 cattle, 811 goats, 58 sheep and 177 pigs in these households used Giemsa-stained blood smears and ITS-PCR amplification for diagnosis of trypanosomiasis. Prevalence was highest in cattle (28.4% [95% CI: 23.7-33.5]) followed by pigs (21.5% [13.9-31.8]), sheep (18.2% [5.1-47.7]), and goats (9.2% [6.8-12.4]). The prevalence within households depended on the particular combinations of livestock species kept; small ruminants were more likely to be infected if cattle were also present. In cattle, prevalence ranged from 26.3% (CI: 19.6-34.2) above 700m above sea level to 44.1% (CI: 36.9-51.6) below 600m. <i>Trypanosoma congolense</i> (savannah type)) was identified in 82.4% of all trypanosome-infected cattle, <i>T. vivax</i> in 24.5%, <i>T. brucei</i> in 2.7%. <i>Trypanosoma simiae</i> was only identified in pigs (27.8% of infected pigs). Finally, ‘restricted application’, a novel modality of use of synthetic pyrethroid to control both tsetse- and tick-borne diseases of cattle was investigated in a longitudinal intervention study in 12 villages in Petauke District on the eastern plateau. Baseline data showed that trypanosomiasis was more prevalent in villages in the northern part of the study are while theileriosis was more prevalent in southern villages. Existing infection of trypanosomes were treated with two doses of diminazene aceturate 42 and 14 days prior to interventions. Spraying only the limbs, belly (predilection feeding sites of tsetse) and ears (predilection feeding site of <i>Rhipicephalus</i> species ticks) of cattle with dilute deltamethrin, was compared with conventional pour-on application of deltamethrin, trypanocidal chemoprophylaxis using isometamidium chloride, and non-intervention controls. Each intervention (or non-intervention) was applied to 80cattle in each of 3 villages, monthly (deltamethrin) or just once (isometamidium). The subsequent incidence of trypanosome infection was too low to make a meaningful conclusion on the interventions. Nevertheless, restricted application had significant effect on tick infestations and animals treated with deltamethrin showed lower cumulative incidence of <i>Theileria </i>species.

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.

Antimicrobial resistance (AMR) is a worldwide public health concern. While it is evident that the use of antibiotics creates selection pressure for the evolution of antibiotic resistance genes, there are still considerable knowledge gaps relating to the status quo of antibiotic use, emergence of resistant pathogens in different livestock production systems and spread within human and animal communities. This thesis includes a survey of antibiotic use in the dairy sector within a specific area of Zambia and analysis of AMR and virulence factors in E. coli isolated from dairy cattle and diarrhoea human patients with the following objectives. 1. To investigate the usage of antibiotics in the dairy sector and the drivers for use. 2. To determine the prevalence and patterns of antimicrobial resistance in E. coli isolated from faecal samples of dairy cattle. 3. To use whole genome sequencing (WGS) to investigate the molecular epidemiology of resistance determinants in E. coli strains isolated from both dairy cattle and humans. 4. To assess the zoonotic potential of isolated E. coli focusing on Shiga toxin-producing E. coli (STEC) and relationship to STEC associated with clinical disease in the UK. In view of these objectives, the first part of the work was carried out in Zambia and involved a questionnaire, a field survey, isolation of E. coli from dairy cattle faecal samples and phenotypic testing for AMR. In addition, E. coli isolates were obtained from another study that was focused on human patients presenting with diarrhoea at the University Teaching Hospital in Lusaka. The second part involved whole genome sequencing and molecular analyses of E. coli for resistance and virulence genotypes at the Roslin Institute (UK). For the field study, a stratified random sample of 104 farms was studied, representing approximately 20% of all dairy farms in the region. On each farm, faecal samples were collected from a random sample of animals and a standardised questionnaire on the usage of antibiotics was completed. An E. coli isolate was obtained from 98.67% (371/376) of the sampled animals and tested for resistance against the six types of antibiotics (tetracycline, ampicillin, sulfamethoxazole/trimethoprim, cefpodoxime, gentamicin and ciprofloxacin). These E. coli were then analysed together with those from humans for genotypes in the laboratory and from Illumina short read whole genome sequences using bioinformatics tools. Tetracylines and penicillin were the commonly used antibiotics in dairy herds. This finding was in line with the resistance phenotypes detected in E. coli isolated from the dairy cattle. The most prevalent AMR was to tetracycline (10.61; 95%CI: 7.40-13.82), followed by ampicillin (6.02; 95%CI: 3.31-8.73), sulfamethoxazole/ trimethoprim (4.49; 95%CI: 2.42-6.56), cefpodoxime (1.91; 95%CI: 0.46-3.36), gentamicin (0.89; 95%CI: 0.06-1.84) and ciprofloxacin (0%). The risk analysis indicated that AMR was associated with livestock diseases (lumpy skin disease and foot rot), exotic breeds (Jersey and Friesian), location, farm size and certain management practices. Analysis of whole genome sequences showed that isolates from humans had both higher levels and a greater diversity of resistance alleles than the cattle isolates. Common genotypes in both populations were: tetA (16%), tetB (10%), tetC (2%) for cattle isolates with tetA (32%), tetB (22%) and tetD (1%) in human isolates. Other common genotypes were blaTEM (56%), sul1 (29%), sul2 (66%), strA4 (57%) and strB1 (64%) in isolates of human origin while blaTEM (15%), sul1 (3%), sul2 (17%), strA4 (13%) and strB1 (19%) were in the cattle isolates. Whilst the E. coli isolates from cattle encoded resistance to common antibiotics of limited significance to human clinical medicine, isolates from humans had additional extended spectrum beta-lactamases (blaOXA, blaCMY, blaNDM, and blaDHA, blaOKP and blaCTX-M) that encode for resistance to essential antibiotics such as third generation cephalosporins and carbapenems. This was an evidence that AMR is an ongoing public health subject in Zambia but the exclusivity of certain resistances in the human population points to limited or no exchange of genotypes between E. coli of human origin and those from cattle. AMR in humans was probably independently selected by the use of antibiotics of clinical importance such as cephalosporin and fluoroquinolones. The virulence analysis focused on STEC, 11% (41/371) of E. coli isolates from cattle contained Shiga toxin genes (stx) while none (0/73) of the human isolates were positive. Phylogenetic analysis showed a random distribution of bovine STEC, with no indication of clonal spread. Although 89% (16/18) of the STEC tested had a cytotoxic effect on Vero cells, indicative of Shiga toxin production, only three (O45, O111, O157) belonged to one of the seven serogroups (O26, O157, O111, O103, O121, O145 and O45) associated with life-threatening enterohaemorrhagic E. coli (EHEC) infections in humans. In line with this, only the O157 serotype encoded a type 3 secretion system. This shows that, while Stx-encoding strains are common in these dairy herds of Zambia, they are not strain backgrounds known to pose an immediate threat to human health as they lack colonisation factors that are found in typical human EHEC. However, we must remain vigilant as emergence of EHEC strains in these animals remains an ever-present threat.

The protozoan parasites Trypanosoma congolense and T. vivax cause trypanosomosis in cattle. The major lysosomal cysteine proteinase of T. congolense, congopain, may contribute to pathogenesis of the disease, and antibody-mediated inhibition of this enzyme may contribute to mechanisms of trypanotolerance. Oligopeptidase B, a trypanosomal serine peptidase, is also a potential virulence factor in African trypanosomes because it is released into the host circulation by dead or dying parasites, where it retains catalytic activity due to the enzyme's insensitivity to serum protease inhibitors. The vaccine potential of the catalytic domain of congopain, C2, and oligopeptidase B complexed with 0'2-macroglobulin (0'2M) was evaluated by producing antibodies in rabbits. Inhibition of congopain and oligopeptidase B activity by these antibodies was assessed. The oligopeptidase B open reading frame from T. congolense and T. vivax was cloned and expressed in Escherichia coli, from which active recombinant enzymes were purified. These recombinant enzymes exhibited trypsin-like specificity for peptide substrates, cleaving on the carboxy side of basic amino acid residues such as arginine and lysine. Enzymes were found to be optimally active between pH 8 and 10, optimally stable at pH 6, and showed activation by reducing agents and sensitivity to ionic strength. The enzymes showed typical oligopeptidase B-like inhibitor profiles, except that they were not inhibited by thiol sensitive inhibitors such as iodoacetamide and Nethylmaleimide. High yields of bovine and rabbit 0'2M were isolated by a three-step procedure of fractionation by PEG 6000, and zinc chelate and Sephacryl S-300 HR chromatography. Congopain, its catalytic domain C2, papain and cathepsin L all cleaved the bait region of bovine 0'2M and became trapped inside the 0'2M molecule, where their activity against large molecular weight substrates was inhibited. C2 could thus be complexed with 0'2M directly or used to form C2-0'2M-oligopeptidaseB complexes for immunisation purposes. iv The catalytic domain of congopain, C2, was used to immunise rabbits either without adjuvant, as a water-in-oil emulsion with Freund's adjuvant, or in a complex with either bovine or rabbit U2M. Freund's adjuvant elicited the highest anti-C2 antibody response. However, the greatest inhibition, 65%, of C2 activity against Z-Phe-Arg-AMC was obtained with antibodies produced by rabbits receiving C2-U2Mcomplexes. In a second study, C2 and oligopeptidase B were used to immunise rabbits , either in alum, or complexed to bovine U2M. Anti-C2 antibody levels were highest in rabbits immunised with the free proteins in alum, whereas anti-oligopeptidase B antibody levels were comparable for each adjuvant system. Anti-oligopeptidase antibodies produced with alum gave 100% inhibition of oligopeptidase B activity. In contrast, antibodies produced against C2-u2M-oligopeptidase B complexes had little effect on oligopeptidase B activity. However, these antibodies inhibited 55% of C2 activity. Alum was a slightly less efficient adjuvant for C2 and 50% inhibition of C2 activity was observed. It appeared that immunisation of rabbits with C2 complexed to U2M resulted in the production of antibodies that were better able to neutralise the proteolytic activity of C2 and congopain in vitro than that with conventional adjuvants . The immunisation of C2 complexed to bovine u2-macroglobulin therefore has the potential to neutralise parasite congopain in vivo, and may contribute to an anti-disease vaccine against African trypanosomosis. Complexation of oligopeptidase B to u2M offers no benefit, since antibodies produced against this complex are not able to inhibit the activity of oligopeptidase B. Immunisation with oligopeptidase B in alum is sufficient to produce efficient enzyme-inhibiting antibodies in the context of an anti-disease vaccine against African trypanosomosis.<br>Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2006.

Trypanosoma congolense is a major causative agent of the bovine disease trypanosomosis which has a considerable economic impact on sub-Saharan Africa. Current control methods for trypanosomosis are unsatisfactory and vaccine development has been hampered by antigenic variation. An anti-disease vaccine is based on the idea that disease is caused by the pathogenic factors released by the parasite, rather than by the parasite itself. Therefore, if these pathogenic factors could be neutralised by antibodies produced by vaccination, the disease could be circumvented. The method used here for identification of novel pathogenic factors is based on the concept that trypanotolerant cattle are able to mitigate the disease by generating a specific immune response against a few key antigens (pathogenic factors). Two immuno-affinity columns were therefore prepared: one containing IgG from noninfected sera and a second column containing IgG from trypanotolerant N’Dama cattle serially infected with T. congolense. The differential binding of antigens to the two columns allowed identification of antigens specifically recognised by the immune system of a trypanotolerant animal, i.e. potential pathogenic factors. The most promising antigens identified included several variant cathepsin L-like cysteine peptidases (CPs) and the Family M1 Clan MA aminopeptidases (APs). For the CPs, a study of the genetic organisation was conducted in order to further understand the variability present in this gene family. To this end, two different mini-libraries of cathepsin L-like genes were prepared: one in which genes as different as possible from congopain (the major CP of T. congolense) were selected, and a second which contained all possible genes present in the congopain array. Analysis of the sequences obtained in these two mini-libraries showed that there was significant variability of the genes within the congopain array. Two variants of CPs, chosen for differences in their catalytic triads, were cloned for expression. The recombinantly expressed CP variants differed in substrate preferences from one another and from C2 (the recombinant truncated form of congopain), and surprisingly, all enzymes were active at physiological pH. The two APs were cloned and expressed as insoluble inclusion bodies in an E. coli system, and subsequently refolded. The refolded APs showed a substrate preference for H-Ala-AMC, an optimum pH of 8.0, localisation to the cytoplasm and inhibition by puromycin. The two APs were not developmentally regulated and present in procyclic, metacyclic and bloodstream form parasites. Down-regulation of both APs by RNAi resulted in a slightly reduced growth rate in procyclic parasites in vitro. Immunisation of BALB/c mice with the APs did not provide protection when challenged with T. congolense. For an anti-disease vaccine to be protective, it would possibly have to include all pathogenic factors, including the two APs and at least one CP described in the present study.<br>Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2010.

Trypanosomosis is a parasitic disease in man, domestic and wild animals and is of major economic importance in many parts of the world, particularly in Sub-Saharan Africa. Trypanosoma congolense, T vivax and T brucei brucei are the major pathogenic trypanosomes infecting cattle in sub-Saharan Africa. The parasite itself is not directly responsible for the disease, but rather causes illness through the release of pathogenic factors. One of the major pathogenic factors released by trypanosomes is proteinases. Trypanotolerant cattle produce antibodies against a trypanosomal proteinase, congopain, that inhibit congopain activity. Congopain thus has vaccine potential. This study describes the mapping of immunogenic epitopes of congopain to identify peptide regions of the protein that induce enzyme inhibitory antibodies for inclusion in a trypanosome vaccine. This vaccine approach targets the disease, rather than the parasite by focusing on a pathogenic factor. These peptides also have potential for use in diagnostic assays. Peptides from the catalytic domain of a trypanosomal cysteine proteinase, congopain, were selected using an epitope prediction program. Peptides selected were from the two forms of congopain called CP1 and CP2. Antibodies against peptide-carrier conjugates were produced in chickens. The antibodies recognised native congopain, recombinant CP2 and the recombinant catalytic domain (C2). This suggests that the peptides selected have promise for use in vaccines. The peptides were also used to determine whether they are natural immunogenic epitopes of CP2 and thus have potential for use in diagnostic assays. Antibodies in the sera from T. congolense infected cattle recognised all the peptides in an ELISA. Antibodies in the sera from C2-immunised, non-infected cattle recognised most of the peptides in an ELISA. In order to distinguish between T. congolense and T vivax infection, two different peptides from the C-terminal extensions of CP2 and vivapain were used in ELISA tests with sera from infected cattle. Although anti-peptide antibodies produced against the two C-terminal extension peptides were specific for their respective peptides, thereby indicating the discriminatory power of the peptides selected, there was cross-reactivity by the sera from T. congolense and T. vivax infected cattle. Optimal antibody binding peptide sequences of these two peptides need to be identified by testing modified sequences of these two peptides to improve the sensitivity of this assay. In addition to attempting to define the epitopes of congopain, preliminary studies to increase the immunogenicity of congopain were also undertaken. Alpha 2-macroglobulin is a natural host inhibitor of proteinases. Inhibition occurs by entrapment of an active proteinase within the alpha 2-macroglobulin cage. In addition, it has been demonstrated that antigen complexed with alpha 2-macroglobulin becomes more immunogenic, resulting in enhanced antigenic presentation of an entrapped antigen. This study reports the interaction between congopain and alpha 2-macroglobulin. The preliminary results of this study showing congopain-alpha 2-macroglobulin interaction could be used to explore the possibility of increasing the immunogenicity of congopain and congopain epitopes by complexing these to alpha 2-macroglobulin. Congopain epitopes complexed with alpha 2-macroglobulin could be used to form a peptide-based vaccine.<br>Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2003.