year 2005 PAAT Programme volume 28 Against African part 1 Trypanosomiasis
TSETSE AND TRYPANOSOMIASIS INFORMATION Numbers 13087–13283
Edited by John N. Pollock Hove, East Sussex United Kingdom of Great Britain and Northern Ireland
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ª'"0 Tsetse and Trypanosomiasis Information
TSETSE AND TRYPANOSOMIASIS INFORMATION
The Tsetse and Trypanosomiasis Information periodical has been established to disseminate current information on all aspects of tsetse and trypanosomiasis research and control to institutions and individuals involved in the problems of African trypanosomiasis. This service forms an integral part of the Programme Against African Trypanosomiasis (PAAT) and is jointly sponsored by the Food and Agriculture Organization of the United Nations (FAO), the International Atomic Energy Agency (IAEA), the Inter-African Bureau for Animal Resources of the African Union (AU-IBAR), the World Health Organization (WHO), the Research Department for Livestock Production and Veterinary Medicine of the Centre de Coopération Internationale en Recherche Agronomique pour le Développement (CIRAD-EMVT), the British Government’s Department for International Development (DFID) and the Institute of Tropical Medicine (ITM), Antwerp.
The half-yearly periodical is prepared for publication, in both English and French editions, by the Food and Agriculture Organization of the United Nations. Each annual volume consists of two parts and an index. Subscription is free for all recipients engaged in trypanosomiasis research and control, and requests for enrolment may be sent to: Ms Maria Grazia Solari, AGAH, FAO, Viale delle Terme di Caracalla, 00100 Rome, Italy (fax +39 06 5705 5749; e-mail [email protected]).
Since the value of this information service depends to a great extent on the receipt of relevant material from research workers, campaign planners and organizers and field workers themselves, readers are requested to submit news items and copies of scientific papers and reports to the Editor: Dr John N. Pollock, 25 Palmeira Mansions, Church Road, Hove, East Sussex, BN3 2FA, United Kingdom (tel. +44 1273 326211; e-mail [email protected]).
We regret that we are unable to supply photocopies of the papers quoted in the periodical.
Distribution dates and copy deadlines
Copy deadline for Distribution news items (English and French editions)
Part 1 15 April July/August Part 2 15 October January/February
The Index will be distributed as soon as possible after the completion of each volume.
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ABBREVIATIONS USED IN TTI a.i. active ingredient LC50 median lethal concentration ACTH adrenocorticotrophic hormone LD50 median lethal dose ALAT alanine aminotransaminase M molar ASAT aspartic acid aminotransaminase mAEC miniature anion-exchange b.w. body weight centrifugation technique BIIT blood incubation infectivity test McAb monoclonal antibody CATT card agglutination test for MW molecular weight trypanosomiasis NARS National Agricultural Research CD50 median curative dose Services/Systems CNS central nervous system p.i. post-infection CSF cerebrospinal fluid PCR polymerase chain reaction DNA deoxyribonucleic acid PCV packed cell volume ELISA enzyme linked immunosorbent assay ppb parts per billion (109) HAT human African trypanosomiasis ppm parts per million HCT haematocrit centrifugation technique r.h. relative humidity GIS geographic information system(s) RNA ribonucleic acid GPS global positioning system(s) SIT sterile insect technique i.m. intramuscular(ly) sp(p). species (plural) i.p. intraperitoneal(ly) ssp(p). subspecies (plural) i.v. intravenous(ly) UV ultra-violet IFAT indirect fluorescent antibody test VAT variable antigen type KIVI kit for in vitro isolation of VSG variant surface glycoprotein trypanosomes WBC white blood cell
Organizations ANDE Agence Nationale de Développement de l’Elevage AU African Union AU/STRC African Union/Scientific, Technical and Research Commission BICOT Biological Control of Tsetse by the Sterile Insect Technique CEBV Communauté Economique du Bétail et de la Viande CEMV Centre Universitaire de Formation en Entomologie Médicale et Vétérinaire CGIAR Consultative Group on International Agricultural Research CIRAD Centre de Coopération Internationale en Recherche Agronomique pour le Développement CIRAD-EMVT Département d’Elevage et de Médecine Vétérinaire des Pays Tropicaux du CIRAD CIRDES Centre International de Recherche-Développement sur l’Elevage en Zone Subhumide CNERV Centre National d’Elevage et de Recherches Vétérinaires CNRS Centre National de Recherche Scientifique CREAT Centre de Recherche et d’Elevage, Avétonou, Togo CRSSA Centre de Recherches du Service de Santé des Armées Emile Pardé CTVM Centre for Tropical Veterinary Medicine DFID Department for International Development (UK) DSE German Foundation for International Development EC/EU European Community/European Union EDF European Development Fund FAO Food and Agriculture Organization of the United Nations FITCA Farming in Tsetse Control Areas of Eastern Africa
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GTZ Deutsche Gesellschaft für Technische Zusammenarbeit IAEA International Atomic Energy Agency IBAR Interafrican Bureau for Animal Resources ICIPE International Centre of Insect Physiology and Ecology ICPTV Integrated Control of Pathogenic Trypanosomes and their Vectors IFAD International Fund for Agricultural Development ILRI International Livestock Research Institute INRA Institut National de Recherche Agronomique IPR Institut Pierre Richet IRD Institut de Recherche et de Développement (formerly ORSTOM) ISCTRC International Scientific Council for Trypanosomiasis Research and Control ISRA Institut Sénégalais de Recherches Agricoles ITC International Trypanotolerance Centre ITM Institute of Tropical Medicine KARI Kenya Agricultural Research Institute KETRI Kenya Trypanosomiasis Research Institute LCV Laboratoire Central Vétérinaire LNERV Laboratoire National de l’Elevage et de Recherches Vétérinaires LSHTM London School of Hygiene and Tropical Medicine MRC Medical Research Council MRU Mano River Union NITR Nigerian Institute for Trypanosomiasis Research NRI Natural Resources Institute OCCGE Organisation de Coopération et de Coordination pour la Lutte contre les Grande Endémies OCEAC Organisation de Coordination pour la Lutte contre les Endémies en Afrique Centrale OGAPROV Office Gabonais pour l’Amélioration de la Production de la Viande OIE Office International des Epizooties OMVG Organisation pour la Mise en Valeur du Fleuve Gambie PAAT Programme against African Trypanosomiasis PATTEC Pan-African Tsetse and Trypanosomiasis Eradication Campaign PRCT Projet de Recherches Cliniques sur la Trypanosomiase RDI Rural Development International RUCA Rijksuniversitair Centrum Antwerpen SADC Southern African Development Community SIDA Swedish International Development Authority SODEPRA Société pour le Développement des Productions Animales TDR UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases TDRC Tropical Diseases Research Centre TPRI Tropical Pesticides Research Institute TTRI Tsetse and Trypanosomiasis Research Institute UNDP United Nations Development Programme USAID United States Agency for International Development USDA United States Department of Agriculture UTRO Uganda Trypanosomiasis Research Organisation WHO World Health Organization
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CONTENTS
Page SECTION A – NEWS Report of the tenth PAG Co-ordinators’ Meeting, Accra 1 Technical Co-operation projects of the Joint FAO/IAEA Division 10
SECTION B – ABSTRACTS 1. General (including land use) 13 2. Tsetse biology (a) Rearing of tsetse flies - (b) Taxonomy, anatomy, physiology, biochemistry 14 (c) Distribution, ecology, behaviour, population studies 20 3. Tsetse control (including environmental side effects) 23 4. Epidemiology: vector-host and vector-parasite interactions 28 5. Human trypanosomiasis (a) Surveillance 33 (b) Pathology and immunology 35 (c) Treatment 40 6. Animal trypanosomiasis (a) Survey and distribution 44 (b) Pathology and immunology 49 (c) Trypanotolerance 51 (d) Treatment - 7. Experimental trypanosomiasis (a) Diagnostics 54 (b) Pathology and immunology 55 (c) Chemotherapeutics 58 8. Trypanosome research (a) Cultivation of trypanosomes - (b) Taxonomy, characterisation of isolates - (c) Life cycle, morphology, biochemical and molecular studies 68
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SECTION A – NEWS
REPORT OF THE TENTH PAAT ADVISORY GROUP CO-ORDINATORS MEETING, ACCRA, GHANA. 22–23 SEPTEMBER 2004
The tenth PAAT Advisory Group (PAG) Coordinators’ meeting was held at the FAO Regional Office for Africa (FAORAF), Accra, Ghana, 22–23 September 2004. The meeting was jointly organized by FAO HQ, Rome and FAORAF, and supported by the Food and Agriculture Organization of the United Nations, the Inter-African Bureau for Animal Resources of the African Union, the International Atomic Energy Agency and World Health Organization of the United Nations. Prof. Albert Ilemobade, Chairman of PAAT, made the introductory remarks, followed by a welcoming statement from Mr Mensah Agyen-Frimpong, Director of Ghana Veterinary Services. Mr Joseph Tchicaya, Assistant Director General/Regional Representative for Africa of FAO officially opened the meeting. Conclusions and recommendations 1. Conclusion: The meeting commended the ADB-PATTEC initiative aimed at financing (through ADB loans and grants) multi-national tsetse and trypanosomiasis (T&T) intervention projects in East (Ethiopia, Kenya and Uganda) and West Africa (Burkina Faso, Ghana and Mali). The PAG appreciated this development and the role of Ghana in bringing together countries concerned to hold the project appraisal workshop in Accra. Recommendation: The meeting: Urges recipient countries to take advantage of available expertise in PAAT for project implementation, monitoring and evaluation; Encourages other governments and their national institutions to show similar levels of commitment to T&T intervention; Urges the strengthening of collaboration between PAAT and PATTEC in bringing about the achievement of the stated goals of the project. Action: ADB beneficiary countries, PAAT and PATTEC. 2. Conclusion: The meeting observed with concern the inability of the FAO Liaison Officers (LO) to perform as required and the dwindling frequency of their meetings. The meeting considers that the LOs play a key role in feeding PAG with information necessary to its work. Recommendation: It is recommended to: Re-activate as a matter of urgency the activities of the FAO LOs; Ensure that terms of reference of the LOs are clear and updated; National Governments to provide the LOs with resources to carry out their tasks. Action: FAO Regional Office for Africa. 3. Conclusion: The meeting noted with appreciation the progress that has been made in the search for new drugs for the treatment of sleeping sickness. The WHO initiative of bridging the gap between research and control by the establishment of
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the HATNETWORK is also acknowledged. This network should facilitate the development of new drugs and the use of new tools. Recommendation: The meeting recommends: To continue to support work on the development of new drugs and new diagnostic tools which can be considered as key elements needed to achieve disease elimination; To form a task force to facilitate and support the implementation of clinical trials for new drugs and the application of new diagnostic tools; To support the development of sub-regional disease elimination programmes. Action: WHO. 4. Conclusion: The meeting recognised the importance of advocacy as a means of accessing resources for the support of T&T interventions and observed that there has been an improvement in the communication between T&T workers and policy makers. Recommendation: The meeting: Urges increasing cooperation and collaboration between policy makers, researchers and field personnel; Recommends that T&T workers include strong advocacy as a key element of their general strategy and to actively participate in: (a) decision making on the elaboration of livestock development policies at the national, sub-regional and continental level; (b) the dissemination of information on T&T, livestock production and rural development; (c) the sensitisation of livestock farmers’ associations and support to these associations and other livestock professionals at the local, national and sub-regional levels. Action: National, regional and international policy makers, CVOs, extension and delivery services. 5. Conclusion: The meeting observed with appreciation the diverse ways in which CIRDES and ITC have supported NARS and institutions charged with T&T interventions in the West African region in the area of training. Also, it was noted with concern the lack of well-defined strategic training needs. Recommendation: The meeting recommends to: Develop mechanisms that would ensure that T&T endemic countries take full advantage of training opportunities; Assist NARS and national institutions charged with T&T interventions to identify strategic training needs; Re-visit PAAT previous recommendations on training needs and where necessary update them in the context of emerging trends. Action: CIRAD, CIRDES, ITM, ITC, PAAT. 6. Conclusion: The meeting recognises the growing importance of public-private sector partnership in national development and in any intervention in T&T. Recommendation: The meeting recommends PAAT to explore avenues for fostering such partnership in the future. Action: PAAT and its stakeholders.
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7. Conclusion: The meeting recognises that the commitment of NARS and other national stakeholders is crucial to the success of T&T interventions. Recommendation: Every effort should be made to stimulate the active participation of NARS and all the stakeholders involved in national and regional T&T intervention programmes. Action: National governments, NARS, National Extension and Delivery Services. 1. Brief and discussion on the last PAG meeting report – A.A. Ilemobade The report of the previous meeting held in Pretoria in September 2003 was discussed, and the conclusions and recommendations endorsed. The PAG was informed that the proposal of the “ICPTV – phase two” was not accepted for funding by the European Union, and ITM was exploring other financial sources. Additional information concerned meetings held in 2004 addressing the standardization of methodologies and devices for tsetse trapping and use of insecticide. In this regard, conclusions and consensus have not been reached yet. The participants agreed on the provisional agenda and time table. The next PAG meeting will be convened in Addis Ababa, Ethiopia, in September 2005 in conjunction with the 28th ISCTRC Conference. 2. The problem of T&T and related control: a move into the future and the importance of PATTEC – K. Gyening Despite a century of efforts, tsetse-transmitted trypanosomiasis still constitutes a major constraint for livestock-agricultural development in sub-Saharan Africa. Main problems in limiting or eliminating the impact of the disease derive from there being only a limited number of drugs available for disease treatment, an increased presence of drug resistant strains of parasites, a reduced capacity of veterinary services in delivering appropriate strategies to farmers for a proper use of chemotherapy, the high cost of vector control campaigns and difficulties in obtaining the involvement and participation of the local communities to support T&T interventions. Elimination of the T&T problem implies its removal and preventing it from coming back: this is a complex exercise. A fundamental prerequisite is the geographic isolation of the vector population(s) which allows the application of integrated area-wide pest management strategy in combination with proper management of land and other natural resources rendering the habitat unsuitable for reinvasion or new vector colonization. Due to the transboundary nature of tsetse distributions, a regional approach is needed. Training and capacity building also have to be considered in the planning phase of a T&T field intervention. Now, for the first time in more than 100 years, the African Nations have collectively shown the political will to free the continent from the burden of T&T. This political will was concretised with the endorsement by the OAU (now AU) of the Pan African Tsetse and Trypanosomiasis Eradication Campaign (PATTEC) in 2000. Currently, ADB has shown interest in supporting T&T interventions through allocation of loans and grants to six tsetse-affected African countries (Burkina Faso, Ethiopia, Ghana, Kenya, Mali and Uganda). Ownership and direct involvement of national governments are necessary to pave the way for success of interventions. In this context, PAAT is likely to provide the ideal environment to establish a mechanism for international collaboration, and neutral, technical advice and assistance in the planning phase. As recommended at the last ISCTRC Executive Committee meeting (Addis
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Ababa, September 2004), further consultation between PAAT and PATTEC is needed for concerted action with regard to this ADB initiative. 3. Report of the PAAT Secretariat and FAO/PAAT activities - R.C. Mattioli In the nine million square kilometres of sub-Saharan (SS) Africa infested by tsetse, 55 percent of the rural population lives below the poverty line. The proportion of under- nourished in sub-Saharan Africa has not dramatically changed since the early nineties. A negative relationship exists between the extent of cultivated area and under-nourishment. For instance, in the Near East and North Africa 28 percent of the total land is cultivated with 10 percent of people undernourished, while in sub-Saharan Africa only 3 percent of the land is cultivated and the fraction of the population undernourished reached 33 percent. Vast areas facing the T&T problem are suitable for livestock-agricultural development. Hence, besides the technical feasibility of any T&T intervention, it is necessary to concentrate efforts in those areas offering potential for land development. PAAT’s strategy is to promote SARD with a view to increase food security and reduce poverty in tsetse affected countries, and the role of PAG, the policy and strategy setting body of PAAT, is to develop standardised normative procedures and methodologies for T&T interventions in the context of SARD. The PAG was informed about FAO and PAAT activities as related to T&T since the 2003 PAG meeting. FAO and PAAT support to member countries and PATTEC was concretised in the endorsement of the FAO Progress Report on the implementation of the PATTEC initiative by the 32nd FAO Conference (Rome, December 2003). The concept note, developed by Ethiopian officials with the assistance of FAO and IAEA, concerning field T&T intervention in the Ethiopian Southern Rift Valley was officially endorsed by the Ethiopian Government. Further actions in the implementation of PAAT-PATTEC agreements concerned the organization of an international workshop on “Strategy for integrated T&T intervention in the ‘cotton-belt’ of Burkina Faso and Mali”, held in Ouagadougou, Burkina Faso, February 2004. Representatives of Burkina Faso, Mali, IAEA and FAO attended the forum. A major output of the workshop was the production of a note of interest concerning “Poverty reduction through reinforcement and intensification of mixed farming in T&T intervention area in the cotton-belt zone of Burkina Faso and Mali”. The note of interest was presented at the 8th PAAT Programme Committee, held in FAO HQ, April 2004. Two PAAT Technical and Scientific Series papers were published, a first paper entitled “Economic guidelines for strategic planning of tsetse and trypanosomiasis control in West Africa”, and a second one dealing with “Long-term tsetse and trypanosomiasis management options in West Africa”. Two additional papers are in press or in progress: (a) The role of trypanotolerant livestock in the context of trypanosomiasis intervention strategies (expected to be published first half of 2005); (b) Mapping the benefits of disease removal as a decision tool for T&T intervention (expected to be published jointly by FAO and DFID second half of 2005). With regard to publications, a joint FAO-IAEA-WHO-ERGO paper was published in the international journal Food, Agriculture and Environment, with the title “Tsetse and trypanosomiasis intervention policies supporting sustainable animal-agricultural development” [see TTI 27(2) 12891]. A radio interview (“Lutte contre la trypanosomose: partir du développement rural”) was broadcast by AGFAX-WREN (World radio for the
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Environment) media in May 2004. A communication on FAO and PAAT activities was presented at the OIE/NTTAT (Non-Tsetse Transmitted Animal Trypanosomiasis) meeting held in Paris, May 2004. In relation to the involvement of the private sector in FAO and PAAT activities, a first draft of the Memorandum of Understanding between FAO and IFAH on the harmonization of standards and protocols for quality control/quality assurance of trypanocides has been produced and is currently under evaluation by the interested parties. Through the FAO Representative in Sudan, preliminary contacts were undertaken with the EU for a possible WHO/FAO project on human and animal trypanosomiasis control in south Sudan. Progress was made in securing a grant from IFAD to support the PAAT Information System (PAAT IS). The grant is expected to be available early 2005. Additional financial support was received from CIRAD-EMVT, ITM and DFID mainly as contributions to the publication of the Tsetse and Trypanosomiasis Information (TTI) bulletin (formerly the Tsetse and Trypanosomiasis Information Quarterly). The PAG was briefed on the outcome of the PAAT Programme Committee (PAATPC) meeting, held in FAO Headquarters, Rome, April 2004. The meeting was attended by representatives of Burkina Faso, Ethiopia, Kenya, Mali, Namibia, AU/IBAR, IAEA, WHO, international research institutes, an FAO-NEPAD related officer, and representatives of the donor community (e.g. ADB, IFAD, Italian Cooperation, USDA/APHIS) and private sector (e.g. IFAH). The meeting focused on (i) progress and achievements of the four PAAT mandated organizations (AU/IBAR, FAO, IAEA, WHO), (ii) advancement in the preparation of conceptual notes for field programme proposals for T&T interventions in the two PAAT-PATTEC agreed priority areas, (iii) economics of T&T intervention, and (iv) the role of the private sector in intervention activities. The report is expected to be available early 2005 and will be distributed through PAAT-Link and will be downloadable from the PAAT website. Future FAO/PAAT activities will concentrate on (i) the advancement of the PAAT- PATTEC harmonization process, (ii) refinement and promotion of common policies for T&T interventions aiming at a positive SARD, (iii) assisting African countries (e.g. those countries receiving ADB support for T&T intervention) in planning SARD related T&T intervention programmes, (iv) implementation of FAO-IFAH partnership on quality control of trypanocides. The PAG was informed about the venue of the next PAAT-PC meeting (IAEA Headquarters, Vienna, 3-4 May 2005) and PAG meeting (Addis Ababa, just prior to the ISCTRC meeting, September 2005). The discussion which followed the presentation drew members’ attention to the inactivity of the FAO Liaison Officers’ (LO) platform. Concern was expressed about the irregularity of the LO meetings. The FAO Regional Office for Africa was urged to re- establish the LO meetings on a regular basis to provide the necessary support for PAG decisions. Some PAG members reported on the inability of the LO to perform efficiently because of logistic constraints. The PAG expressed agreement on the need for training, essential in the preparation for the implementation of field T&T intervention activities. In this regard, the meeting agreed that there is a general concern about a vacuum being created at the middle level category/personnel due to the retirement (and ageing) of most
5 2005 Tsetse and Trypanosomiasis Information of those who were trained in the seventies and eighties. The open door policy of CIRDES and ITC to the training needs of countries in the West African region was commended. 4. Report of AU/IBAR activities – J. Musiime The AU/IBAR report focused on implementation and achievements of the FITCA project, which is now operative in five East African countries: Ethiopia, Kenya, Rwanda, Tanzania and Uganda. The overall objective aims at the improvement of the welfare of people through sustainable rural development. A major specific objective concerns increased livestock-agricultural productivity by improving animal health through T&T control and integration of crop-livestock production with a view to increase level of food security. A key element of the FITCA project is the promotion of partnerships between local communities, public and private sectors and NGOs. In this regard, local communities are the primary agents in supporting tsetse control activities. Village communities are mainly active in the deployment and management of insecticide impregnated targets. In addition, farmers’ communities (more than 600 farmers’ groups established in Kenya and Uganda) engage in building and managing “crush pens” where cattle are treated epicutaneously with insecticides (upon payment up to KShs20), dewormed and receive additional health treatments. In fact, “crush pens” are now used as Rural Animal Health Centres by the village communities assisted by private animal health providers. Work carried out by FITCA has shown that tsetse control and rural development are interdependent. On the one hand, without controlling flies farming cannot become more productive; on the other hand, improvement of the rural infrastructure is hampered where agriculture is unproductive due to the presence of trypanosomiasis. Where the trypanosomiasis constraints were removed (i.e. in certain project areas in Kenya, Uganda, Ethiopia and Tanzania) or tsetse fly controlled, integrated crop-livestock farming systems were established. The introduction of insecticide-treated netting in Kenya, Uganda and Tanzania appears effective in protecting zero-grazing cattle production units not only from tsetse flies, but also from other nuisance biting insects, including mosquitoes. The demand for impregnated nettings is increasing and AU/IBAR is assisting in acquiring official registration of the netting technique for its easy availability on the market in Kenya and Uganda. Cases of sleeping sickness have been dramatically reduced in project areas in Kenya and Uganda. In the latter country, medical teams are fully operational in 12 districts and several “sleeping sickness hospitals” have been renovated in Uganda. A monitoring, surveillance, diagnosis and treatment scheme has been established and free treatments have been made available through WHO/private sector (AVENTIS) co- operation. In the project area, the Environment Management and Monitoring Component (EMMC) of FITCA is implemented through a special agreement with ILRI and the Scientific Environment Monitoring Group. A baseline survey on the environment has been carried out in Kenya, Uganda and Ethiopia using remote satellite imagery backed by ground-truthing in 3–4 selected villages of 5 km2 in each study area. The field survey assessed natural vegetation and biodiversity, current land use and, where possible, land use changes over time. This information is complemented with socio-economic surveys. FITCA financed research activities involving several national (KETRI, Kenya; LIRI,
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Uganda) and international (ILRI, ICIPE) institutions. The total amount of research fund allocated was €200 000. Finally, the presenter informed the house that the next ISCTRC Conference will be held 26th September–1st October 2005, in Addis Ababa, Ethiopia. 5. Report from IAEA – U. Feldmann (based on a note sent to the PAAT Secretariat) Since the last PAG meeting (September 2003, Pretoria, South Africa), the IAEA’s tsetse activities underwent a major internal and external review evaluation. While the findings and conclusions are not available at this stage, it is likely that the evaluation will have implications for the future interaction of the Agency with member states and other partners and stakeholders in the field of tsetse and trypanosomiasis. The Agency’s objective is to promote the use of nuclear techniques in the context of sustainable development. Although disputed by some, the sterile insect technique (SIT) is generally recognised as a proven technique when applied within an area-wide integrated pest management (AW-IPM) strategy. As such, the SIT component has the potential to contribute to the removal of the T&T problem in some selected development areas in African member states. The Agency’s mandate and specific expertise and experience lie with the transfer of SIT in support of this goal. Therefore, its support activities will, in the future, be largely restricted to the SIT component of AW-IPM. Other activities that were previously also supported by IAEA, such as conventional suppression activities using insecticides on livestock or artificial baits prior to the SIT phase, will, in future, likely need to be conducted by other partners or stakeholders. The Agency’s assistance to member states efforts against T&T will continue to benefit from its scientific-technical and management expertise. Substantially increased emphasis will be on the principles of national and regional ownership and international harmonization and prioritization. The Agency recognises that attaining the creation of tsetse-free zones will require a concerted action by many partners over many years, including appropriate policy, institutional and technological interventions with substantial human resources and financial commitments. IAEA will, therefore, work with development agencies and other partners to assist its member states in enhancing the effectiveness of their activities by strengthening their human resources and infrastructure and leveraging the financial contributions required to ensure that the full potential of SIT can be realised as a contribution to the wider development goal and in the context of the AU-PATTEC initiative. Further harmonization between PATTEC and PAAT is expected to avoid duplication of efforts by the different international partners and other stakeholders and to provide synergetic inputs to member states in support of the common development objective. Relevant services to member states will continue to be conducted using both the Agency’s Regular Budget (RB) resources and the IAEA Technical Cooperation Funds (TCF). RB funds will continue to be used for: Normative work, including the development of standards, guidelines, manuals, etc.) in close collaboration with other partners and through mechanisms like PAAT; In-house research at the FAO/IAEA Agriculture and Biotechnology Laboratory, Seibersdorf, Austria; Coordinated Research Projects (CRPs); and
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Consultants’ meetings on relevant topics. With regard to TCF, the Agency consistently experiences increasing member states’ demand for TC support activities in the field of T&T, and it remains a challenge to secure sufficient resources to meet the demand. In the near future, it is intended to pursue ways of enhancing the impact of TC projects, through better up-front planning, increased monitoring of results and expanded partnerships. In this context, the Agency’s tsetse related activities obviously will reflect and be in line with member states’ national development plans and priorities, and technical assistance to field projects will follow a phased, conditional approach. After a screening of the national policies, priorities and commitment and an initial phase of baseline data collection, the subsequent phases (feasibility assessment, pre- operational phase and operational phase) would only be supported by the Agency provided pre-agreed conditions/criteria are met. In all phases, the Agency would only contribute to assist in the SIT-relevant component. With regard to IAEA TCF supported activities in member states, IAEA currently supports SIT-baseline data collection, feasibility assessment and capacity building in ten T&T affected countries through eight national and one regional projects. As SIT is only a component of larger projects for creating tsetse-free zones and for overall sustainable agriculture and rural development, and considering the complex and multidisciplinary nature of such operations, IAEA does not and will not attempt to manage or lead such field programmes in the member states. The Agency’s role is to provide relevant guidance and advice on the establishment of appropriate national/sub-regional management structures for the effective execution of AW-IPM projects with an SIT component. Consistent with the efforts initiated under PAAT-PATTEC harmonization process and the above mentioned phased, conditional approach, the Agency will need to focus on a limited number of operational projects with the potential to create tsetse-free zones, provide capacity building where feasibility study was positive, and continue to support requests for the feasibility studies in other member states as appropriate. Regarding new member states’ request to IAEA for technical assistance in the field of T&T, the Agency will need to introduce a more rigorous project appraisal process, with attention being paid to the government’s commitment, the assessed technical feasibility, the relevance and anticipated impact of the proposed collaborative efforts in the context of sustainable agriculture and rural development, and the availability of Agency resources (human and financial) to ensure the effective delivery of its input. 6. Report from WHO – J. Jannin Since the establishment of PAAT, WHO has adapted its strategy to meet PAAT’s overall objectives, namely to provide a networking platform, to collect and disseminate information, to develop policy and strategy and to promote advocacy and awareness. These actions resulted in the establishment of a strong partnership with all those involved in sleeping sickness surveillance and control. These objectives complement the WHO’s mandate for coordination, support and resource mobilization for field activities and research. In practical terms WHO has created a forum where people can not only exchange ideas and share views on the final goal to eliminate sleeping sickness but also propose new activities and approaches to improve sleeping sickness surveillance and control. This forum took the name of the Human African Trypanosomiasis Network and brings together everyone concerned through a networking concept. Within this
8 2005 Tsetse and Trypanosomiasis Information networking process much progress has taken place and has been regularly reported at the PAAT meetings. Awareness has been considerably raised among the decision makers and the population at risk. With the willingness of public agencies and the private companies, coherent actions could be implemented. It is now conceivable and there is a great hope that elimination of sleeping sickness, as public health problem, will be achievable. The times when some 60 000 cases were diagnosed and treated while only 10 to 15 percent of the population at risk were under some surveillance is over. In the last years there has been a regular decline of cases, and in 2004 the number of new cases was less than 17 000, while there has been a substantial increase in the number of people under surveillance. The most outstanding achievements were an improvement in diagnostic quality, a substantial increase in the use of eflornithine as first line treatment and the development of the short melarsoprol treatment protocol, which has undergone a full clinical trial as a preliminary step before for any recommendation as an appropriate approach for treating patients. One could also mention the new molecule DB289 which is presently undergoing field trials. This new chemical is a promising alternative for first stage treatment. Although research is still needed, it is likely that it will also be adapted for the treatment of late stage sleeping sickness cases. Trials using nifurtimox and eflornithine combination are ongoing. Drug combination is considered a promising approach as an alternative therapy to avoid resistance. Furthermore, new molecules are under consideration and novel diagnostic tests are being validated. One of these tests is a highly specific and sensitive serological assay while another is an agglutination test to assess the involvement of the nervous central system. Other diagnostic tools are under consideration and there is good hope for their further development. Thus, networking has allowed the development, validation and introduction of new techniques in restructured and reinforced national programmes, which continue to receive seeding financial support to enhance surveillance and control. Training continues to provide the essential competence to implement the new tools as they become available. Certainly there are still countries where surveillance and control are still weak but improvements are constantly being made. It is unlikely that the disease will ever be eradicated, due to the nature of its reservoir, but it will undoubtedly be eliminated as public health problem. It is now time to look at the approaches and methodologies to sustain the results obtained. It seems timely to identify tools that can be integrated into health delivery systems to maintain surveillance. Sleeping sickness has been and occasionally still is an obstacle to rural development but it will soon be time to look at other diseases that represent stumbling blocks. This 10th PAAT Advisory Group Coordinators’ meeting should certainly take note of the recent success which is the result of organized determination, strong collaboration and networking. Everyone here should be satisfied to have brought a building stone to the edifice. In that context, FAO has progressed towards the improvement of meat and milk production and towards a better, more productive and sustainable agriculture, leading to rural development and food security in T&T affected areas. Together, lead agencies are moving more efficiently towards the ultimate goal of an improved economic and social status of the African population.
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[Due to lack of space the following sections are presented here by the titles and speakers only, but are due to appear soon in full in the report of the PAG meeting on the main AGA webpage as well as on the PAAT website (see www.fao.org/ag/paat.html)] 7. Progress on the refinement of the concept note for Burkina Faso and Mali: information available and information needed – I. Sidibe 8. Concerted action: the role and contribution of international research institutes (CIRAD, CIRDES, ITC, ITM) in supporting T&T intervention and related research activities – S. de la Rocque, I. Sidibe, K. Agyemang, S. Geerts 9. Concerted action: the role and contribution of NARS in T&T intervention and related activities – V. Codjia, I. Tamboura, C. Mahama, E. Coulibaly, C. Ly 10. Advancing together: from Burkina Faso, Mali and Ethiopia experience, the role of PAAT in assisting African countries in defining national T&T/SARD strategies as related to proposal(s) for joint international intervention – Moderators P.Holmes, A.A. Ilemobade 11. Report on advancement of FAO-IFAH partnership for Quality Control/Quality Assurance of trypanocides – R.C. Mattioli 12. Identification of areas for additional public-private sector partnership and support – Moderator V. Codjia.
TECHNICAL CO-OPERATION PROJECTS OF THE JOINT FAO/IAEA DIVISION
The following items of interest to the readership of TTI are summarised from Insect Pest Control Newsletter, No. 64, published by the Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture and FAO/IAEA Agriculture and Biotechnology Laboratory, Seibersdorf, Austria.
Ongoing Technical Co-operation Project: Feasibility Studies for Area-wide Intervention in Mali
The suitability of an area for intervention against its tsetse infestation depends on several factors. One is the extent of the tsetse infestation, considered on a species-by-species basis. Another is the possibility of re-invasion of cleared areas by flies from outside the cleared zone; a critical issue here is the efficiency of natural barriers such as watersheds between river basins to prevent re-infestation. Lastly, it needs to be confirmed that the control method under review actually works in the particular study area. The TCP project has addressed several of these factors. A survey of Glossina palpalis gambiensis distribution was carried out in the northern part of the Niger River Basin. After study of previous survey results, a zone of more than 9 000 km2 was surveyed. The new results indicated that most flies were taken from the vegetation of the left bank of the Niger River, and the northern limit to the present-day distribution appears to be south of that found in the earlier surveys. There
10 2005 Tsetse and Trypanosomiasis Information was some evidence of the very fragmented nature of the tsetse population distribution in this northern zone that had been predicted from theoretical studies. The dispersal potential of the G. palpalis gambiensis populations was assessed by means of an analysis of the gene frequencies in these populations, sampling from various river tributaries. Very little genetic heterogeneity was detected in the mitochondrial DNA and genetic diversity was low, indicating past genetic bottlenecks. As there was little genetic differentiation between tsetse populations in the various tributaries, it was concluded that the respective tsetse populations were not well isolated. Larger-scale sampling is called for to amplify these preliminary results. The above genetic study was complemented by a release-recapture exercise. The small-scale experiment conducted using colony-reared G. p. gambiensis indicated that flies could disperse over a watershed from the Niger catchment to the Senegal Basin. Such results lead us to approach the Area-Wide Integrated Pest Management strategy with extreme caution in this instance. Locally, tsetse (Glossina palpalis gambiensis) has been drastically suppressed using insecticide-impregnated Vavoua traps in the extended peri-urban area around Bamako. The intervention zone is to be increased, but in future it is expected that the first zone will be monitored and action taken at points where the fly is beginning to recover in numbers.
Developments affecting tsetse rearing at the Entomology Unit, Seibersdorf
A semi-automated system for rearing flies (TBU3) is being installed at Seibersdorf. The idea is to reduce further still the stress imposed on the breeding female tsetse flies, to encourage maximum productivity. The new unit also allows greater ease of management, so that one or two technicians can deal with a colony of more than a million females. It was anticipated that the new installation would be completed by January 2005. Efforts are being made to automate the sexing of tsetse pupae, using a near infra- red spectrometer. After the delivery of the spectrometer, tests will be carried out on Glossina pallidipes pupae, to separate them automatically into males and females. If these trials are satisfactory, the technique will be extended to other species. Some basic research into sound production in tsetse is being conducted. The pattern of sound production, the frequency spectrum of the sounds, the effects of time of day on calling are some of the areas being investigated. It is hoped that studying the sounds produced by tsetse will give information concerning factors important in successful mating, and help towards making a general index of fly fitness useful in colony quality control. Possible improvements to the quality of the blood fed to the fly colonies are being studied. Pasteurization is a possible alternative to irradiation to reduce bacteria in the blood feed. Various anti-coagulants are being tested, as well as other additives such as dyes, and trials are under way to test the advantages of different ratios of amino acids and lipids. The frequency and effect of re-mating in female tsetse flies has been studied in G. pallidipes and G. brevipalpis. In G. pallidipes, a low rate of re-mating (0–10 percent) was observed. It was felt that such levels were unlikely to have a significant effect on an SIT programme as sterilised males will be released in very high numbers compared to the
11 2005 Tsetse and Trypanosomiasis Information flies present in the field. In G. brevipalpis, a small genetic study gave preliminary indications that where a female mated twice, the second-mating male’s sperm were used for fertilizing the eggs. More observations are needed here.
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SECTION B - ABSTRACTS
1. GENERAL (INCLUDING LAND USE)
13087 Anon., 2004. African trypanosomiasis or sleeping sickness – fact-sheet. Weekly Epidemiological Record, 79 (32): 297–300.
WHO: Avenue Appia 20, 1211 Geneva 27, Switzerland.
A broad introduction to the disease is given, together with estimates of the geographical extent and the size of the population affected; the reader is warned that the true incidence is grossly underestimated by official statistics.
13088 Buguet, A., Bouteille, B., Cespuglio, R., Bisser, S., Chapotot, F., Bourdon, L., Vincendeau, P., Radomski, M.W. & Dumas, M., 2003. Maladie du sommeil: la recherche oubliée? [Sleeping sickness: has research been forgotten?] Médecine Tropicale, 63 (3): 223–227.
Buguet: Institut de médecine tropicale du service de santé des armées, Le Pharo, B.P. 46, 13998 Marseille Armées, France.
To investigate whether HAT has been forgotten as a target for research, a bibliographic database was searched for reports concerning studies in this area. It was found that the number of publications on HAT had remained steady, whereas those on malaria had shown a steady increase. Over 14 years, 1 million Euros had been spent to produce 68 publications recorded in this database.
13089 Iyayi, E.A., Okoruwa, V.O., Babayemi, O.J., Busari, A.A. & Peters, O.F., 2003. Livestock production pattern of agropastoralists in peri-urban centres of south-west Nigeria. Nigerian Journal of Animal Production, 30 (1–2): 87–92.
Peters: Department of Animal Science, University of Ibadan, Ibadan, Nigeria.
The remarkable reduction of trypanosomiasis and its vector, the tsetse fly, in the South West zone of Nigeria has led to the development of agropastoralism in the zone. This present study was carried out by the administration of structured questionnaires to farmers in three towns (Oyo, Ogbomosho and Saki) in order to highlight some of the factors influencing production in the area. It was found that in all the three towns, animals were maintained on free range grazing, browsing and feeding of crop residues. The most favoured breed kept is the Bunaji, with the highest concentration found in Ogbomosho (52 percent). Labour allocation among agropastoralists was based on sex. Diarrhoea was the most prevalent disease among the adult animals in wet and dry seasons while sand eating was common among the calves. In all the centres, cattle constituted the major
13 2005 Tsetse and Trypanosomiasis Information 1 ruminant in stock (77 percent) while sheep and goats accounted for 15 and 8 percent, respectively. A preponderance of female cattle over the male for all the breeds was recorded in all the towns. Saki had the highest number of cattle in stock and Oyo the lowest. Feed supplements offered all year round were salt lick and grains. Most of the agropastoralists depended on the use of local herbs and other such methods of combating diseases affecting the herd.
13090 Jannin, J. & Cattand, P., 2004. Treatment and control of human African trypanosomiasis. Current Opinion in Infectious Diseases, 17 (6): 565–571.
Access to treatment is a multi-step process and little progress has been made to improve treatments for sleeping sickness over the past 50 years. The current strategy is based on diagnostic tools developed in the 1960s while available drugs are still the same as those developed in the middle of the last century. Strategic opportunities can only be based on two achievements: improved diagnosis and safer drugs. This paper reviews the development of new diagnostic tools and drugs and the opportunity offered by new technologies for their further improvement. The prodrug DB289 shows excellent oral activity with low toxicity for the treatment of early-stage sleeping sickness; it has recently entered phase II(b) clinical trials. The recent ability to identify and test specific host and parasite biomarkers has allowed the development of new diagnostic and stage- determination tools that are more specific and sensitive. The accurate diagnosis of an infection by use of proteomic signature analysis has been achieved. Urinary nitrites and nitrates follow closely the increase of brain nitric oxide associated with the penetration of trypanosomes in the brain. Sleep-onset rapid eye movement-like episodes have been shown to occur at onset of late-stage trypanosomiasis. This unique disturbance of the wake/sleep cycle seems to be the first pathognomonic sign in the occurrence of late-stage trypanosomiasis. Following the description of the disease, and diagnostic tools and drugs that have been used, and are still in use today, the authors show how it has influenced over time the evolution of strategies for surveillance and control. Recent developments and prospects for new, more specific and sensitive diagnostic tools and a safer drug will undoubtedly improve the accuracy of patient recruitment and facilitate treatment, and provide opportunities for new strategies.
2. TSETSE BIOLOGY
(a) REARING OF TSETSE FLIES
(b) TAXONOMY, ANATOMY, PHYSIOLOGY, BIOCHEMISTRY
13091 Abila, P.P., Kiendrebeogo, M., Mutika, G.N., Parker, A.G. & Robinson, A.S., 2003. The effect of age on the mating competitiveness of male Glossina fuscipes fuscipes and G. palpalis palpalis. Journal of Insect Science, 3 (13): 1– 8.
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Abila: Livestock Health Research Institute (LIRI), P.O. Box 96, Tororo, Uganda. [[email protected]]
The effect of age on male Glossina fuscipes fuscipes and Glossina palpalis palpalis competitiveness were investigated with a view to estimate optimal age for sterile male release. Sterile insect technique involves the mass production, sterilization and sequential release of males of the target species to out compete the wild male population. Mating between released sterile males and wild females produce inviable progeny and the population is reduced over several generations to unsustainable levels. It is vital that the released male be of high quality and are sexually competitive. Age is one parameter affecting the sexual competitiveness of the male tsetse fly. The optimal release age was estimated by assessing sexual competitiveness of flies of different age categories, 1, 5, 8 and 13 days after adult eclosion. A walk-in field-cage was used in order to approximate as closely as possible the actual field scenario during sterile insect release programes. It was shown that 8- and 13-day old males mated significantly more frequently, i.e. were more competitive, in the presence of equal numbers of 1- and 5-day old males. The age of male tsetse flies significantly affected competitiveness in both species studied. The ability of G. f. fuscipes to inseminate was not age dependent, and insemination occurred in all females that mated regardless of male age. In G. p. palpalis, however, 1-day old males were least able to inseminate. Mating duration was not significantly affected by age in both species. Eight to thirteen days are here recommended as the optimal age for males for the sterile male release in the test species.
13092 Abubakar, L.U., Zimba, G., Wells, C., Mulaa, F. & Osir, E.O., 2003. Evidence for the involvement of a tsetse midgut lectin-trypsin complex in differentiation of bloodstream-form trypanosomes. Insect Science and its Application, 23 (3): 197–205.
Osir: International Centre of Insect Physiology and Ecology, P. O. Box 30772, Nairobi, Kenya.
A bloodmeal-induced molecule (lectin-trypsin complex) from the midgut of the tsetse fly, Glossina longipennis, with both lectin and trypsin activities, has been previously described. In this paper, the isolation of a similar molecule is reported from the midguts of Glossina fuscipes fuscipes and direct evidence for its involvement in the development of African trypanosomes is provided. The molecule (native Mr ~65,700) has two non-covalently linked subunits, Mr ~28,800 and Mr ~35,700. The native molecule was found to be capable of inducing differentiation of bloodstream-form trypanosomes into procyclic (midgut forms) in vitro. In the assays, specific antibodies against procyclin were used to monitor the transformation of the bloodstream-form trypanosomes into procyclic forms. This induction was specifically inhibited by D-glucosamine. Cis- aconitate was also capable of inducing the transformation process with the same efficiency as that of the lectin-trypsin complex. While increasing the concentrations of the lectin-trypsin complex in the incubation assays resulted in higher transformation rates, it also led to high parasite mortality. These results provide evidence for the
15 2005 Tsetse and Trypanosomiasis Information 2b involvement of the midgut lectin-trypsin complex in the differentiation of bloodstream- form trypanosomes within the tsetse midgut.
13093 Aksoy, S., Berriman, M., Hall, N., Hattori, M., Hide, W. & Lehane, M.J., 2005. A case for a Glossina genome project. Trends in Parasitology, 21 (3): 107–111.
Aksoy: Yale University School of Medicine, 60 College Street, 606 LEPH, New Haven, CT 06520, USA.
Given the medical and agricultural significance of Glossina, knowledge of the genomic aspects of the vector and vector-pathogen interactions has a high priority. In preparation for a full genome sequence initiative, an extensive set of expressed sequence tags (ESTs) has been generated from tissue-specific normalised libraries. In addition, bacterial artificial chromosome (BAC) libraries are being constructed, and information on the genome structure and size from different species has been obtained. An international consortium is now in place to further efforts to lead to a full genome project.
13094 Cabrero, P., Pollock, V.P., Davies, S.A. & Dow, J.A.T., 2004. A conserved domain of alkaline phosphatase expression in the Malpighian tubules of dipteran insects. Journal of Experimental Biology, 207 (19): 3299–3305.
Dow: Division of Molecular Genetics, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G11 6NU, UK. [j.a.t.dow@ bio.gla.ac.uk]
Malpighian (renal) tubules are key components of the insect osmoregulatory system and show correspondingly great diversity in both number and length. Recently, the organization of the Drosophila melanogaster tubule has been elucidated by enhancer trapping, and a gene expression array for functional properties has been shown to align with the functional domains. In Drosophila, there is a lower tubule domain, which coincides with expression of alkaline phosphatase and delineates the absorptive region of the tubule. Here, these observations are extended to three dipteran disease vectors (Aedes aegypti, Anopheles stephensi and Glossina morsitans) and a non-dipteran out-group, Schistocerca gregaria. Despite a huge range in cell number and size, alkaline phosphatase was found on the apical surface of the lower 10 percent of each of the dipteran tubules but nowhere within the orthopteran tubule. An alkaline phosphatase lower tubule domain is thus conserved among Diptera. Cell counts were also provided for each species. As in Drosophila, stellate cells were not found in the lower tubule domain of Anopheles or Aedes tubules, confirming the unique genetic identity of this domain. As previously reported, we failed to find stellate cells in Schistocerca but, remarkably, also failed to find them in Glossina, the dipteran most closely related to Drosophila. The orthodoxy that stellate cells are unique to, and general among, Diptera may thus require revision.
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13095 Dale, C., Jones, T. & Pontes, M., 2005. Degenerative evolution and functional diversification of type-III secretion systems in the insect endosymbiont Sodalis glossinidius. Molecular Biology and Evolution, 22 (3): 758–766.
Dale: Department of Biology, University of Utah, Salt Lake City, USA. [[email protected]]
Sodalis glossinidius, a maternally transmitted endosymbiont of tsetse flies, maintains two phylogenetically distinct type-III secretion systems encoded by chromosomal symbiosis regions designated SSR-1 and SSR-2. Although both symbiosis regions are closely related to extant pathogenicity islands with similar gene inventories, SSR-2 has undergone novel degenerative adaptations in the transition to mutualism. Notably, SSR-2 lacks homologues of genes found in SSR-1 that encode secreted effector proteins known to facilitate the host cell cytoskeletal rearrangements necessary for bacterial entry and uptake into eukaryotic cells. Also, as a result of relaxed selection, SSR-2 has undergone inactivation of genes encoding components of the type-III secretion system needle substructure. In the current study, we used quantitative PCR to determine the expression profiles of ysaV (SSR-1) and invA (SSR-2) transcripts when S. glossinidius infects an insect cell line, and we used an invasion assay to characterise the phenotype of an S. glossinidius mutant that lacks the ability to produce an OrgA protein that is required for function of the SSR-2 secretome. Whereas SSR-1 is required for bacterial invasion of host cells and ysaV is expressed when bacteria contact host cells, SSR-2 is required for bacterial proliferation after entry, and invA is only expressed in the intracellular stage of infection. These results demonstrate that degenerative genetic adaptations in SSR-2 have promoted functional diversification of the Sodalis SSR-2 type-III secretion system.
13096 Dinglasan, R.R., Valenzuela, J.G. & Azad, A.F., 2005. Sugar epitopes as potential universal disease transmission blocking targets. Insect Biochemistry and Molecular Biology, 35 (1): 1–10.
Dinglasan: Department of Microbiology and Immunology, University of Maryland School of Medicine, 20 Penn Street, Baltimore, MD 21201, USA.
One promising method to prevent vector-borne diseases is through the use of transmission blocking vaccines (TBVs). However, developing several anti-pathogen TBVs may be impractical. In this study, we have identified a conserved candidate carbohydrate target in the midguts of several arthropod vectors. Tests suggested that the anticarbohydrate malaria transmission blocking monoclonal antibody (MG96) preferentially recognises a particular discontinuous glycotope on arthropod vector midguts. The data suggest that these glycotopes may represent potential transmission blocking vaccine targets for a wide range of vector-borne pathogens.
13097 Gooding, R.H. & Krafsur, E.S., 2005. Tsetse genetics: Contributions to biology, systematics, and control of tsetse flies. Annual Review of Entomology, 50: 101–123.
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Gooding: Department of Biological Sciences, University of Alberta, Edmonton, Alberta T6G 2E9, Canada.
This review covers the distribution and importance of tsetse flies; the tsetse life cycle; population biology; the status of tsetse genetics; genetic variation; linkage groups; sex determination; population genetics; systematics and genetic methods of population suppression. In conclusion, it is mentioned that our knowledge of the genetics of tsetse greatly lags behind that of the pathogenic organism Trypanosoma.
13098 Gooding, R.H., Solano, P. & Ravel, S., 2004. X-chromosome mapping experiments suggest occurrence of cryptic species in the tsetse fly Glossina palpalis palpalis. Canadian Journal of Zoology – Revue Canadienne de Zoologie, 82 (12): 1902–1909.
Gooding: Univ Alberta, Department of Biological Sciences, University of Alberts, Edmonton, AB T6G 2E9, Canada.
Using flies from colonies of Glossina palpalis palpalis that originated in Nigeria and Bas-Zaire, the two microsatellite loci Gpg19.62 and Gpg55.3 have been added to the X-chromosome map, thus increasing to seven the number of loci mapped on that chromosome. During the mapping and other crossing experiments, sterile F1 and backcross males were found. Similarities between the patterns of sterility found in the present study and those occurring during hybridization of some subspecies of tsetse suggest that the nominal taxon G. p. palpalis may contain cryptic taxa. Differences in the width of the postgonite head in the genitalia of males from the two colonies were consistent with this suggestion.
13099 Hens, K., Macours, N., Claeys, I., Francis, C. & Huybrechts, R., 2004. Cloning and expression of the yolk protein of the tsetse fly Glossina morsitans morsitans. Insect Biochemistry and Molecular Biology, 34 (12): 1281–1287.
Hens: Department of Biology, Catholic University of Leuven, Naamsestraat 59, 3000 Leuven, Belgium.
Two major families of nutritional proteins exist in insects, namely the vitellogenins and the yolk proteins. While in other insects only vitellogenins are found, cyclorraphan flies only contain yolk proteins. Possible sites of yolk protein synthesis are the fat body and the follicle cells surrounding the oocyte. We report the cloning of the yolk protein of the tsetse fly Glossina morsitans morsitans, a species with adenotrophic viviparity. The tsetse fly yolk protein could be aligned with other dipteran yolk proteins and with some vertebrate lipases. In contrast to the situation in most fly species, only a single yolk protein gene was found in the tsetse fly. Northern blot analysis showed that only the ovarian follicle cells, and not the fat body, represent the site of yolk protein synthesis.
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13100 Hu, Y.J. & Aksoy, S., 2005. An antimicrobial peptide with trypanocidal activity characterised from Glossina morsitans morsitans. Insect Biochemistry and Molecular Biology, 35 (2): 105–115.
Aksoy: Department of Epidemiology and Public Health, Section of Vector Biology, Yale University, School of Medicine, 60 College St., 606 LEPH, New Haven, CT 06510, USA
Tsetse flies are vectors of African trypanosomes, the protozoan agents of devastating diseases in humans and animals. Prior studies in trypanosome infected Glossina morsitans morsitans have shown induced expression and synthesis of several antimicrobial peptides in fat body tissue. Here, we have expressed one of these peptides, Attacin (GmAttA1) in Drosophila (S2) cells in vitro. We show that the purified recombinant protein (recGmAttA1) has strong antimicrobial activity against Escherichia coli-K12, but not against the enteric gram-negative symbiont of tsetse, Sodalis glossinidius. The recGmAttA1 also demonstrated inhibitory effects against both the mammalian bloodstream form and the insect stage Trypanosoma brucei in vitro (minimal inhibitory concentration MIC50 0.075 ȝM). When blood meals were supplemented with purified recGmAttA1 during the course of parasite infection, the prevalence of trypanosome infections in tsetse midgut was significantly reduced. Feeding fertile females GmAttA1 did not affect the fecundity or the longevity of mothers, nor did it affect the hatchability of their offspring. We discuss a paratransgenic strategy, which involves the expression of trypanocidal molecules such as recGmAttA1 in the midgut symbiont Sodalis in vivo to reduce trypanosome transmission.
13101 Manosree Chandra, Liniger, M., Tetley, L., Roditi, I. & Barry, J.D., 2004. TsetseEP, a gut protein from the tsetse Glossina morsitans, is related to a major surface glycoprotein of trypanosomes transmitted by the fly and to the products of a Drosophila gene family. Insect Biochemistry and Molecular Biology, 34 (11): 1163–1173.
Barry: Wellcome Centre for Molecular Parasitology, University of Glasgow, 56 Dumbarton Road, Glasgow G11 6NU, UK.
African trypanosomes live in the lumen of the gut of tsetse (Glossina) and may have to face an immune response. As yet, it is unclear whether they are sensitive to antimicrobial peptides in vivo, but for some years there has been indirect evidence that one or more lectins can influence the infection. We have purified a protein complex from midgut extracts that, by SDS-PAGE, is a doublet of 37 and 38 kDa in a ratio of 3:1. Through prediction from corresponding cDNA clones, the full-length protein (tsetseEP) contains 320 amino acids, including a signal peptide. There is apparently only one gene encoding this protein. Towards the C terminus, the protein contains a run of 59 (EP) repeats, which surprisingly is what comprises almost the entire mature EP procyclin molecule present on the surface of trypanosomes in the tsetse gut. Drosophila contains a number of genes encoding proteins, of unknown function, with the same cysteine pattern as tsetseEP; this pattern is not reported for any other protein. Immunoblotting with a
19 2005 Tsetse and Trypanosomiasis Information 2c monoclonal antibody against (EP) repeats reveals expression in the gut, but not salivary glands, of female and male flies, whether or not fed. Immunoelectron microscopy shows the presence in vesicles in midgut cells and in the lumen of the gut. Attempts to demonstrate lectin activity were thwarted by limited availability of the protein complex.
13102 Pollock, V.P., McGettigan, J., Cabrero, P., Maudlin, I.M., Dow, J.A.T. & Davies, S.A., 2004. Conservation of capa peptide-induced nitric oxide signalling in Diptera. Journal of Experimental Biology, 207 (23): 4135–4145.
Davies: Institute of Biomedical and Life Sciences, Division of Molecular Genetics, University of Glasgow, Glasgow G11 6NU, UK. [[email protected]]
In Drosophila melanogaster Malpighian (renal) tubules, the capa peptides stimulate the production of nitric oxide (NO) and guanosine 3ƍ, 5ƍ-cyclic monophosphate (cGMP), resulting in increased fluid transport. The roles of NO synthase (NOS), NO and cGMP in capa peptide signalling were tested in several other insect species of medical relevance within the Diptera (Aedes aegypti, Anopheles stephensi and Glossina morsitans) and in one orthopteran out-group, Schistocerca gregaria. NOS immunoreactivity was detectable by immunocytochemistry in tubules from all species studied. Drosophila melanogaster, Ae. aegypti and An. stephensi only expressed NOS in the principal cells, whereas G. morsitans and S. gregaria showed a general NOS expression in the tubule. Measurement of associated NOS activity (NADPH diaphorase) showed that both D. melanogaster capa-1 and the 2 capa peptides encoded in the An. gambiae genome, QGLVPFPRVamide (AngCAPA-QGL) and GPTVGLFAFPRVamide (AngCAPA-GPT), all stimulated NOS activity in D. melanogaster, Ae. aegypti, An. stephensi and G. morsitans tubules but not in S. gregaria. Furthermore, capa-stimulated NOS activity in all the Diptera was inhibited by the NOS inhibitor L-NAME. All capa peptides stimulated an increase in cGMP content across the dipteran species, but not in S. gregaria. Similarly, all capa peptides tested stimulated the fluid secretion in D. melanogaster, Ae. aegypti, An. stephensi and G. morsitans tubules, but had either no effect or were inhibitory on S. gregaria. Consistent with these results, the Drosophila capa receptor was shown to be expressed in Drosophila tubules, and its closest Anopheles homologue was shown to be expressed in Anopheles tubules. This was the first demonstration of the physiological roles of two putative An. gambiae neuropeptides. We also demonstrated neuropeptide modulation of fluid secretion in tsetse tubule for the first time. Finally, we show the generality of capa peptide action, to stimulate NO/cGMP signalling and increase fluid transport, across the Diptera, but not in an orthopteran.
(c) DISTRIBUTION, ECOLOGY, BEHAVIOUR, POPULATION STUDIES
[See also 28: 13097]
13103 Barrozo, R., Schilman P., Minoli, S. & Lazzari, C., 2004. Daily rhythms in disease-vector insects. Biological Rhythm Research, 35 (1–2): 79–92.
20 2005 Tsetse and Trypanosomiasis Information 2c
Barrozo: Laboratorio de Fisiología de Insectos, DBBE, Facultad de Ciencias, Exactas y Naturales Universidad de Buenos Aires, Argentina
The host-vector-parasite interaction offers a clear illustration of the adaptive value of biological rhythms. In this review, we summarise some of the information currently available on daily rhythms of insect vectors, particularly those responsible for the transmission of parasites to humans. Included amongst the cases described are circadian rhythms of locomotor and flight activity, and of eclosion and oviposition in tsetse flies, blood-sucking hemipterans, mosquitoes and other haematophagous insects. Both published and new data are presented, and they indicate that a study of the rhythms in disease-vector insects can provide an understanding of the value of this application of chronobiology to ecology and applied sciences. Related to this, the possibility of strategies for the control of insects based on their temporal characteristics is proposed; for example, the use of insecticides could be restricted to those phases of the day when the susceptibility of the insects to them is increased.
13104 Dankwa, D., Oddoye, E.O.K. & Mzamo, K.B., 2000. Efficacy of three trap types for trapping Glossina palpalis, G. tachinoides, and G. mortisans in the Mole Game Reserve in Ghana. Ghana Journal of Agricultural Science, 33 (2): 177–181.
Dankwa: Animal Research Institute, Council for Scientific and Industrial Research, P. O. Box AH 20, Achimota, Ghana.
A study was undertaken to determine the most effective of three trap types (biconical, monoconical, and NGU Siamese (NG2B)) for catching three Glossina species (G. palpalis, G. tachinoides, and G. mortisans). The three species were trapped in the Mole Game Reserve. Results showed that in the absence of odour attractants, the biconical trap was the most efficient trap. Out of a total of 4 619 tsetse flies caught during the trapping period, 2 073 (44.9 percent) were caught in the biconical trap. The monoconical trap caught 1 362 tsetse flies (29.5 percent) whilst the NG2B trap caught 1 184 tsetse flies (25.6 percent). There were, however, locational differences, with the number of flies caught in the riverine area being significantly higher than the number caught in the pond area, but not significantly different from the number caught in the woodland. There was also a very significant difference in the species of tsetse flies caught by the traps. Significantly more G. tachinoides were caught as compared to G. mortisans; virtually no G. palpalis were caught. The number of female flies caught (3 621) was significantly higher than the number of male flies caught (995).
13105 Geerts, S., Mahama, C.I., Kone, A., de la Rocque, S., & De Deken, R. 2005. Prediction of the distribution of Glossina tachinoides (Diptera: Glossinidae) in the Volta basin of northern Ghana. Bulletin of Entomological Research, 95 (1): 63–67.
21 2005 Tsetse and Trypanosomiasis Information 2c
Geerts: Animal Health Department, Institute of Tropical Medicine, Nationalestraat 155, B-2000, Antwerp, Belgium.
The classification of a Landsat Thematic Mapper satellite image helped demonstrate prevailing habitat types and land use intensity in the Volta basin of the Northern Region of Ghana. A geo-referenced data layer comprising the capture results of a cross-sectional survey of Glossina tachinoides was overlaid on a data layer of habitat types within 500m of either bank of the Volta River and its tributaries. An evaluation of the relationship between habitat types and the capture results of G. tachinoides suggested a strong preference of G. tachinoides for woodland, followed by shrubland, grassland and flood plains. The findings were used to classify the suitability of habitat types for G. tachinoides as ‘high’, ‘medium’ and ‘low’ and a prediction map for the distribution of G. tachinoides in the entire river network was produced. The usefulness of this method in estimating the potential distribution of G. tachinoides in an area of increasing agricultural expansion is discussed.
13106 Getachew Abebe, Malone, J.B. & Thompson, A.R., 2004. Geospatial forecast model for tsetse-transmitted animal trypanosomosis in Ethiopia. SINET: Ethiopian Journal of Science, 27 (1): 1–8.
Thompson: Faculty of Veterinary Medicine, Addis Ababa University, PO Box 34, Debre Zeit, Ethiopia.
The aims of this study were to develop a geographic information system (GIS) forecast and risk assessment model for cyclically transmitted trypanosomosis in Ethiopia by matching features in the environment to available information on the preferences and limits of tolerance of the parasite and its vector, and validate and further develop the GIS model using data from current and historical prevalence survey data and habitat microenvironment studies from trypanosomosis endemic areas. Results indicate that a GIS model developed for parasitic diseases based on the growing degree day (GDD) concept can be applied to tsetse-transmitted trypanosomosis. These GIS for animal trypanosomosis were created using Food and Agriculture Organization-Crop Production System Zones (FAO-CPSZ) database and Normalised Difference Vegetation Index (NDVI) and maximum temperature (Tmax) from the Global Land 1 km dataset. The two datasets used to determine the risk of tsetse flies and consequently animal trypanosomosis in Ethiopia indicated the magnitude of the disease and possible expansion of the “fly belt” in the future. The GIS model indicated the distribution and importance of tsetse- transmitted trypanosomosis in Ethiopia. Spatial and statistical analyses verified the use of the GDD concept and climate forecast system that were developed to determine the suitability of a given environment for the proliferation of a biotype, in this case tsetse and trypanosomes. Results presented here indicate the importance of GDD and the two climatic variables (NDVI and Tmax) in the development of a forecast model for tsetse- transmitted trypanosomosis in Ethiopia.
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13107 Marquez, J.G., Vreysen, M.J.B., Robinson, A.S., Bado, S. & Krafsur, E.S., 2004. Mitochondrial diversity analysis of Glossina palpalis gambiensis from Mali and Senegal. Medical and Veterinary Entomology, 18 (3): 288–295.
Krafsur: Department of Entomology, Iowa State University, Ames, IA 50011, USA.
West African riverine tsetse populations of Glossina palpalis gambiensis were investigated for gene flow, inferred from mitochondrial diversity in samples of 69 flies from Senegal and 303 flies from three river drainages in Mali. Four polymorphic mitochondrial loci were scored. Mean haplotype diversities were 0.30 in Mali, 0 in Senegal and 0.18 over both Mali and Senegal. These diversities estimate the probabilities that two randomly chosen tsetse have different haplotypes. Substantial rates of gene flow were detected among flies sampled along tributaries belonging to the river basins of the Senegal, Niger, and Bani in Mali. There was virtually no gene flow between tsetse in Senegal and Mali. No seasonal effects on gene flow were detected. The implications of these preliminary findings for the implementation of area-wide integrated pest management (AW-IPM) programmes against riverine tsetse in West Africa are discussed.
3. TSETSE CONTROL (INCLUDING ENVIRONMENTAL SIDE EFFECTS)
[See also 28: 13091, 13097]
13108 Admassu, B., Nega, S., Haile, T., Abera, B., Hussein, A. & Catley, A., 2005. Impact assessment of a community-based animal health project in Dollo Ado and Dollo Bay Districts, Southern Ethiopia. Tropical Animal Health and Production, 37 (1): 33–48.
Catley: African Union/Interafrican Bureau for Animal Resources, PO Box 30786, 00100 Nairobi, Kenya.
Participatory methods were used to assess the impact of a community-based animal health worker (CAHW) project in two remote pastoralist districts of Ethiopia. The CAHW project had been operating for three years at the time of the assessment. Participatory methods were standardised and repeated with ten groups of informants in the project area. The assessment showed significant reductions in disease impact for diseases handled by CAHWs compared with diseases not handled by CAHWs. In camels, there was significant reduction in the impact of mange, trypanosomosis, helminthosis, anthrax and non-specific respiratory disease. In cattle there was a significant reduction in the impact of blackleg, anthrax and helminthosis. In sheep and goats there was a significant reduction in the impact of mange, helminthosis, contagious caprine pleuropneumonia, orf and non-specific diarrhoea. In order of importance, these reductions in disease impact were attributed to (1) increased use of modern veterinary services provided by CAHWs, (2) vaccination campaigns involving CAHWs, (3) good rainfall and availability of grazing and (4) decreased herd mobility. Decreased herd
23 2005 Tsetse and Trypanosomiasis Information 2c,3 mobility was also associated with negative impact of tick infestation. Community-based animal health workers were considered to be highly accessible, available, affordable and trustworthy relative to other service providers. They were also perceived to be suppliers of a good quality service. Specific types of positive impact attributed to CAHW activities were increases in milk, meat, income and draught power.
13109 Barclay, H.J. & Hargrove, J.W., 2005. Probability models to facilitate a declaration of pest-free status, with special reference to tsetse (Diptera: Glossinidae). Bulletin of Entomological Research, 95 (1): 1–11.
Barclay: Pacific Forestry Centre, 506 West Burnside Road, Victoria, British Columbia V8Z 1M5, Canada.
A methodology is presented to facilitate a declaration that an area is ‘pest-free’ following an eradication campaign against an insect pest. This involves probability models to assess null trapping results and also growth models to help verify, following a waiting period, that pests were not present when control was stopped. Two probability models are developed to calculate the probability of negative trapping results if in fact insects were present. If this probability is sufficiently low, then the hypothesis that insects are present is rejected. The models depend on knowledge of the efficiency and the area of attractiveness of the traps. To verify the results of the probability model, a waiting period is required to see if a rebound occurs. If an incipient but non-detectable population remains after control measures are discontinued, then a rebound should occur. Using a growth model, the rate of increase of an insect population is examined starting from one gravid female or one male and a female. An example is given for tsetse in which both means and confidence limits are calculated for a period of 24 reproductive periods after control is terminated. If no rebound is detected, then a declaration of eradication can be made.
13110 De Deken, R., Speybroeck, N., Gillain, G., Sigue, H., Batawi, K. & Van den Bossche, P., 2004. The macrocyclic lactone “spinosad”, a promising insecticide for tsetse fly control. Journal of Medical Entomology, 41 (5): 814– 818.
De Deken: Veterinary Department, Institute of Tropical Medicine, Nationalestraat 155, B-2000, Antwerp, Belgium.
The susceptibility of tsetse flies, Glossina palpalis gambiensis, and G. m. morsitans to topically applied spinosad, a mixture of insecticidal molecules from the actinomycete Saccharopolyspora spinosa, is almost as high as to deltamethrin. However, susceptibility to spinosad does not differ significantly between teneral and gravid flies, contrary to the reaction to deltamethrin. Spinosad might be a promising candidate for future tsetse control by the sequential aerial technique.
24 2005 Tsetse and Trypanosomiasis Information 3
13111 Foil, L.D., Coleman, P., Eisler, M., Fragoso-Sanchez, H., Garcia-Vazquez, Z., Guerrero, F.D., Jonsson, N.N., Langstaff, I.G., Li, A.Y., Machila, N., Miller, R.J., Morton, J., Pruett, J.H. & Torr, S., 2004. Factors that influence the prevalence of acaricide resistance and tick-borne diseases. Veterinary Parasitology, 125 (1–2): 163–181.
Foil: Entomology Department, 402 Life Sciences Building, Louisiana State University Agricultural Center, Baton Rouge, LA 70803, USA.
Within a broader survey, this review of acaricide resistance mentions that the use of pesticides for the control of pests of cattle other than ticks can affect the incidence of tick resistance and tick-borne diseases. Simple analytical models of tick- and tsetse-borne diseases suggest that reducing the abundance of ticks, for example by treating cattle with pyrethroids, can have a variety of effects on tick-borne diseases. In the worst-case scenario, the models suggest that treating cattle might not only have no impact on trypanosomosis but could increase the incidence of tick-borne disease. In the best-case scenario, treatment could reduce the incidence of both trypanosomosis and tick-borne diseases.
13112 Habtewold, T., Prior, A., Torr, S.J. & Gibson, G., 2004. Could insecticide- treated cattle reduce Afrotropical malaria transmission? Effects of deltamethrin-treated Zebu on Anopheles arabiensis behaviour and survival in Ethiopia. Medical and Veterinary Entomology, 18 (4): 408–417.
Gibson: FARM-Africa, Southampton Place, London, UK.
Anopheles arabiensis is the most widespread vector of malaria in the Afrotropical Region. Because An. arabiensis feeds readily on cattle as well as humans, the insecticide- treatment of cattle as employed to control tsetse and ticks might simultaneously affect the malaria vectorial capacity of this mosquito. Field experiments in southern Ethiopia were conducted to establish whether Zebu cattle treated with a pour-on pyrethroid formulation of 1 percent deltamethrin, widely used to control ticks and tsetse, would be effective against An. arabiensis or cause the female mosquitoes to feed more frequently on humans due to behavioural avoidance of insecticide-treated cattle. Contact bioassays (3 min exposure) showed that the insecticide remained effective for about 1 month (kill rate > 50 percent) against mosquitoes feeding on the flanks of treated cattle. A novel behavioural assay demonstrated that An. arabiensis readily fed on insecticide-treated cattle and were not deflected to human hosts in the presence of treated cattle. DNA-fingerprinting of bloodmeals revealed that in nature An. arabiensis feeds most frequently on older animals, coinciding with the established practice of applying insecticide only to older cattle, in order to allow younger untreated animals to gain immunity against infections transmitted by ticks. These encouraging results were tempered by finding that >90 percent of An. arabiensis, An. pharoensis and An. tenebrosus females feed on the legs of cattle, farthest from the site of pour-on application along the animal’s back and where the treatment may be least residual due to weathering. Observations of mosquitoes feeding naturally on insecticide-treated cattle showed that the majority of wild female anophelines alighted on
25 2005 Tsetse and Trypanosomiasis Information 3 the host animal for less than 1 min to feed, with significantly shorter mean duration of feeding bouts on insecticide-treated animals, and the effective life of the insecticide was only 1 week. Thus the monthly application of deltamethrin to cattle, typically used to control tsetse and ticks, is unlikely to be effective against An. arabiensis populations or their vectorial capacity. Even so, it seems likely that far greater impact on anopheline mosquitoes could be achieved by applying insecticide selectively to the legs of cattle.
13113 Hargrove, J.W., 2005. Extinction probabilities and times to extinction for populations of tsetse flies Glossina spp. (Diptera: Glossinidae) subjected to various control measures. Bulletin of Entomological Research, 95 (1): 13–21.
Hargrove: 9 Monmouth Rd, Avondale, Harare, Zimbabwe
A stochastic branching process was used to derive equations for the mean and variance of the probability of, and time to, extinction in tsetse populations. If the remnant population is a single inseminated female, the extinction probability increases linearly with adult mortality and is always certain if this mortality >3.5 percent per day even for zero pupal mortality. If the latter mortality is 4 percent per day, certain extinction is only avoided if adult mortality <1.5 percent per day. For remnant female populations >1, the extinction probability increases in a non-linear manner with adult mortality. Extinction is still certain for adult mortality >3.5 percent per day but, when the remnant population is >16, extinction is highly unlikely for adult mortality <2.5 percent per day if all females are inseminated. Extinction probability increases with increasing probability of sterile mating in much the same way as it does with increasing adult mortality. Extinction is assured if the probability of insemination can be reduced to 0.1. The required reduction decreases with increasing adult mortality. For adult mortality equal to 6–8 percent per day, the time to extinction increases only by one generation per order of magnitude increase in the starting population. Time to extinction is less sensitive to changes in the pupal than in the adult mortality. Reductions in the probability of insemination only become important when adult mortality is small; if the adult mortality is 8 percent per day, reducing the insemination probability from 1 to 0.1 only reduces the expected time to extinction by two generations. Conversely, increases in adult mortality produce important reductions in the required time even when the probability of insemination is 0.1. The practical, economic implication for the sterile insect technique is that the low- tech methods used to suppress tsetse populations should not be halted when the release of sterile males is initiated. The sterile insect technique should only be contemplated when it has been demonstrated that the low-tech methods have failed to effect eradication. The theory is shown to be in good accord with the observed results of tsetse control campaigns involving the use of odour-baited targets in Zimbabwe and the sterile insect technique on Unguja Island, Zanzibar.
13114 Muriuki, G.W., Njoka, T.J., Reid, R.S. & Nyariki, D.M., 2005. Tsetse control and land-use change in Lambwe valley, south-western Kenya. Agriculture Ecosystems and Environment, 106 (1): 99–107.
26 2005 Tsetse and Trypanosomiasis Information 3
Muriuki: Kenya Trypanosomiasis Research Institute, PO Box 362, Kikuyu, Kenya.
For a long time, trypanosomosis spread by the tsetse fly constrained human settlement in the Lambwe Valley, a south-western Kenya rangeland. After lengthy efforts to control tsetse over many years, the valley is currently experiencing an increase in human population growth rate, and rapid changes in land-use and cover are taking place. Using time-series aerial photograph interpretation, social survey methods, and a review of human population trends over five decades, a three-fold expansion in cultivation in the settled areas over a 50-year period, with a consequent decrease in woody vegetation cover was identified. In the Ruma National Park, occupying a third of the valley floor, shrublands and thickets have expanded while open grasslands have decreased. The sudden increase of land under cultivation adjacent to prime agricultural land designated for wildlife conservation, exacerbated by bush encroachment and dwindling resources for tsetse control could provide a situation suitable for land-use conflicts. Sustainability of this unique rangeland is dependent on how judiciously the resources are shared among all stakeholders in the valley. This study suggests continued tsetse surveillance and agricultural intensification in the settled areas to minimise chances of conflicts in land- use.
13115 Otim, C.P., Ocaido, M., Okuna, N.M., Erume, J., Ssekitto, C., Wafula, R.Z.O., Kakaire, D., Walubengo, J., Okello, A., Mugisha, A. & Monrad, J., 2004. Disease and vector constraints affecting cattle production in pastoral communities of Ssembabule district, Uganda. Livestock Research for Rural Development, 16 (5): article 35.
Monrad: Royal Veterinary and Agricultural University, Dyrlaegevej 100, DK-1870 Frederiksberg C, Denmark. [[email protected]]
A survey was carried out to determine the disease and vector constraints to cattle production in pastoral communities settled in three villages of two sub-counties in Ssembabule district, Uganda. A total of 454 cattle randomly selected were sampled for blood parasite infections (antibody ELISA serology and direct parasite detection), for gastrointestinal parasites and for tick infestation. Tsetse exposure was assessed by trapping. The prevalence of trypanosomiasis was low on direct trypanosome detection in blood samples (0.7 percent) as well as on serology (6.7 percent), and no tsetse flies were encountered. Three head of cattle (0.7 percent) were infected with Theileria parva, the cause of East Coast Fever (ECF). However, serological analysis for antibodies against Theileria parva revealed a very high prevalence of exposed animals (93.3 percent). Three head of cattle (0.7 percent) were further found infected with Anaplasma marginale. In addition, 59.1 percent of the cattle examined were shedding worm eggs. Meanwhile 28.7 percent of the sera of cattle tested using Rose Bengal rapid agglutination test were positive for bovine brucellosis. A relatively high tick infestation was demonstrated. Rhipicephalus appendiculatus was found to be the most abundant species, followed by R. evertsi and Amblyomma variegatum. In a preceding questionnaire survey the cattle owners in Ssembabule district had pointed to trypanosomiasis as their most important
27 2005 Tsetse and Trypanosomiasis Information 3 disease problem. This laboratory-based diagnostic survey, however, revealed that ticks, tick-borne diseases and brucellosis were the real major constraints to cattle production in that district, while tsetse and trypanosomiasis appeared to be of little or no importance. It is concluded that planning of disease control measures in a given area should depend on laboratory-based diagnostic observations rather than on a questionnaire survey.
4. EPIDEMIOLOGY: VECTOR-HOST AND VECTOR-PARASITE INTERACTIONS
[See also 28: 13100, 13132, 13147, 13149]
13116 Abenga, J.N. & Lawal, I.A., 2005. Implicating roles of animal reservoir hosts in the resurgence of Gambian trypanosomosis (sleeping sickness). African Journal of Biotechnology, 4 (2): 134–137.
Abenga: Pathology, Epidemiology and Statistics Division, Nigerian Institute for Trypanosomiasis Research, PMB 2077, Kaduna, Kaduna State, Nigeria. [[email protected]]
Gambian trypanosomosis is a complex and debilitating disease of man. For many years the disease has been ravaging several parts of sub-Saharan Africa despite decades of therapeutic control. Although animal reservoir hosts are believed to be associated with the disease, not much evidence has been brought forward to prove the existence of animal reservoir hosts for Trypanosoma brucei gambiense and the zoonotic nature of Gambian sleeping sickness. This paper reviews recent evidence based on molecular and other biotechnologies leading to the identification of mammalian hosts as reservoirs of T. b. gambiense and the roles of such hosts in the transmission and resurgence of sleeping sickness in sub-Saharan Africa.
13117 Chiejina, S.N., Musongong, G.A., Fakae, B.B., Behnke, J.M., Ngongeh, L.A. & Wakelin, D., 2005. The modulatory influence of Trypanosoma brucei on challenge infection with Haemonchus contortus in Nigerian West African Dwarf goats segregated into weak and strong responders to the nematode. Veterinary Parasitology, 128 (1–2): 29–40.
Behnke: School of Biology, University of Nottingham, Nottingham NG7 2RD, UK
Although Nigerian West African Dwarf (WAD) goats are relatively resistant to infection with Haemonchus contortus and are also trypanotolerant, natural outbreaks of both infections are known to occur. Despite their relative resistance to H. contortus WAD goats nevertheless show variability in response phenotype and it was of interest to examine the effect of this variability on the outcome of concurrent trypanosome infection. Trypanosoma brucei infections were established in goats that were initially classified as good or poor responders to H. contortus. Thirty-nine goats were exposed to an escalating
28 2005 Tsetse and Trypanosomiasis Information 4 infection with H. contortus, and on the basis of their mean faecal egg counts (FEC) were allocated to high FEC (poor responders, 18 goats with the highest FEC) or low FEC (good responders, 18 goats with the lowest FEC) classes. Nine uninfected naïve control goats were included to provide reference baseline values. Retrospective analysis of parasitological and pathological parameters after allocation into high/low FEC classes showed that FECs differed significantly, in both classes packed cell volume (PCV) values fell relative to naïve controls, neither class lost weight and both generated marked IgG responses. All goats received anthelminthic on day 61, half of each group was infected with 50 million trypanosomes and on day 67, excepting the controls, all goats were challenged with 3000 L3 of H. contortus. Trypanosome parasitaemia was generally low, and marginally, but not significantly, higher in the low compared with high FEC class, peaking 12–16 days after exposure in both groups and then falling to below microscopically detectable levels (although still detectable by sub-inoculation into mice) by week 3. At autopsy (days 109–110), worm burdens were significantly higher in the trypanosome-infected goats from the high FEC class, relative to all other groups. Trypanosome infected goats showed a tendency (although not significant) towards higher FEC and, irrespective of their FEC class, had lower PCV values although body weight did not vary significantly. All goats challenged with H. contortus had higher antibody levels than naïve controls, but neither trypanosome infection nor FEC class affected the magnitude of responses. These results confirm that WAD goats comprise a range of response phenotypes to initial H. contortus infection and that trypanotolerance is a key trait of this breed. Although immunity to nematode infection develops even in poor responders, these animals harbour higher nematode burdens during concurrent infection with T. brucei.
13118 Chretien, J.-P.L. & Smoak, B.L., 2005. African trypanosomiasis: changing epidemiology and consequences. Current Infectious Disease Reports, 7 (1): 54–60.
Chretien: Department of Defense Global Emerging Infections Surveillance & Response System (DoD-GEIS), Division of Preventive Medicine, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, MD 20910, USA.
Human African trypanosomiasis has re-emerged as a serious public health threat after near-elimination because of diminished investment in previously successful control programs. The continued, occasional importation of African trypanosomiasis to the United States can be expected as tourists and immigrants travel from high-risk areas. No vaccine or chemoprophylaxis is available for this disease, and travellers to affected areas should be counselled on tsetse fly avoidance. New diagnostic and staging tests are promising but have not replaced the classical method of examining body fluids for trypanosomes. Prompt diagnosis and staging is essential because if untreated, East African and West African sleeping sickness are fatal. Drug regimens are toxic and cumbersome, and short-term prospects for therapeutic advances are limited.
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13119 Courtin, F., Dupont, S., Zeze, D.G., Jamonneau, V., Sané, B., Coulibaly, B., Cuny, G. & Solano, P., 2005. Trypanosomose Humaine Africaine: transmission urbaine dans le foyer de Bonon (Côte d’Ivoire). [Human African trypanosomiasis: urban transmission in the Bonon (Côte d’Ivoire) focus.] Tropical Medicine and International Health, 10 (4): 340–346.
Solano: Institut Pierre Richet, Rue Fleming zone 4C, Abidjan 04, Côte d’Ivoire.
Human African trypanosomiasis (HAT) is a vector-borne parasitic disease which has often been considered a rural disease. Population increases in African countries have entailed the spread of urban centres, creating favourable conditions for the appearance of new epidemiological conditions. In Côte d’Ivoire, HAT transmission has been described in the surroundings of towns such as Daloa or Sinfra. In the focus of Bonon, located in central-western Côte d’Ivoire, a medical survey detected 96 patients. The sites visited by the patients every day were geo-referenced and the routes between them recorded. In parallel, an entomological survey of the patients’ daily locations enabled the collection of data on the vector. In Bonon, we observed urban cases and tsetse (Glossina palpalis) feeding on people. Trypanosoma brucei gambiense was identified in both man and vector; thus all conditions for possible intra-urban trypanosomosis transmission were met. The consequences of this are discussed regarding the problem of diffusion of the disease.
13120 Grébaut, P., Mbida, J.A.M., Kondjio, C.A., Njiokou, F., Penchenier, L. & Laveissière, C., 2004. Spatial and temporal patterns of human African trypanosomosis (HAT) transmission risk in the Bipindi focus, in the forest zone of Southern Cameroon. Vector Borne and Zoonotic Diseases, 4 (3): 230– 238.
Grébaut: Laboratoire de Recherche et de Coordination contre les Trypanosomoses (LRCT) CIRAD-IRD, Montpellier, France.
Vector control is an effective and cost-efficient way to disrupt the transmission of human African trypanosomosis (HAT); it has nonetheless been little used to date in the disease foci. With the aim to target trapping more precisely and to develop an optimised vector control system, a transmission risk index was used in the HAT focus of Bipindi, in the forest zone of southern Cameroon. The authors used a simplified version of the index originally developed by Laveissière et al. in 1994. The calculation of this new index only requires knowledge of the proportion of teneral flies and the proportion of flies with human blood meals in samples caught in different biotopes. This makes it possible to identify the biotopes displaying permanent risk, such as riverbanks, as well as biotopes displaying seasonal risk, such as marshy hollows and encampments. In the villages, the domestic pig, with 49 percent of the identified blood meals, is the favourite host of the tsetse flies during the short rainy season. The proportion of blood meals taken on human beings does not significantly increase when domestic pigs are absent. Game animals, contributing to 46 percent and 64 percent of the blood meals during the short rainy season
30 2005 Tsetse and Trypanosomiasis Information 4 and the long dry season, respectively, are also favoured as feeding hosts in this particular HAT focus.
13121 Jamonneau, V., Ravel, S., Koffi, M., Kaba, D., Zeze, D.G., Ndri, L., Sane, B., Coulibaly, B., Cuny, G. & Solano, P., 2004. Mixed infections of trypanosomes in tsetse and pigs and their epidemiological significance in a sleeping sickness focus of Côte d’Ivoire. Parasitology, 129 (6): 693–702.
Solano: Institut Pierre Richet, s/c Représentation IRD, Rue Chevalier de Clieu, 15 BP 917, Abidjan 15, France. [[email protected]]
In a sleeping sickness focus of Côte d’Ivoire, trypanosomes were characterised in humans, pigs and tsetse using various techniques. Out of 74 patients, all the 43 stocks isolated by KIVI (Kit for In Vitro Isolation) appeared to belong to only one zymodeme of Trypanosoma brucei gambiense group 1 (the major zymodeme Z3). The only stock isolated on rodents belonged to a different, new, zymodeme (Z50), of T. b. gambiense group 1. From 18 pigs sampled in the same locations as the patients, PCR showed a high proportion of mixed infections of T. brucei s.l. and T. congolense riverine-forest. Zymodemes of T. brucei s.l. from these pigs were different from those found in humans. From a total of 16 260 captured tsetse (Glossina palpalis palpalis), 1 701 were dissected and 28 percent were found to be infected by trypanosomes. The most prevalent trypanosome was T. congolense riverine-forest type, followed by T. vivax, T. brucei s.l. and T. congolense savannah type, this latter being associated with the forest type of T. congolense in most cases. Mixed infections by two or three of these trypanosomes were also found. Use of a microsatellite marker allowed us to distinguish T. b. gambiense group I in some of the mature infections in tsetse. Differences in infection rates and in trypanosome genotypes according to the host might indicate that the pig may not be an active animal reservoir for humans in this focus.
13122 Magona, J.W., Walubengo, J., Odiit, M., Okedi, L.A., Abila, P., Katabazi, B.K., Gidudu, A.M. & Olaho-Mukani, W., 2005. Implications of the re- invasion of Southeast Uganda by Glossina pallidipes on the epidemiology of bovine trypanosomosis. Veterinary Parasitology, 128 (1–2): 1–9.
Magona: Livestock Health Research Institute, POBox 96, Tororo, Uganda.
A study to assess the influence of re-invasion of Glossina pallidipes on the epidemiology of bovine trypanosomosis was conducted in Southeast Uganda. A total of 1 992 cattle were screened for trypanosomosis in villages using a combination of the BCT and HCT methods; of these cattle, 949 head were in villages infested with G. pallidipes and 1 043 head were in villages without that species. The prevalence of trypanosomosis (15.5 vs 7.1 percent), Trypanosoma brucei infection (1.4 vs 0.6 percent), T. congolense infection (7.2 vs 2.0 percent), T. vivax infection (5.3 vs 3.3 percent) and mixed infection (1.6 vs 1.2 percent) in cattle in villages with G. pallidipes was significantly higher than in those without G. pallidipes. Infections of Trypanosoma congolense were predominant in cattle in villages with G. pallidipes, while T. vivax
31 2005 Tsetse and Trypanosomiasis Information 4 infections were predominant in cattle in villages without that species. In all villages, T. brucei infections were fewer than either T. congolense or T. vivax infections. The risk of transmission of T. brucei, T. congolense and T. vivax infections was 3, 2.7 and 1.6 times higher, respectively, in villages with G. pallidipes than in those without that species, despite the presence of G. f. fuscipes in both sets of villages. The mean PCV and mean herd size of cattle in villages with G. pallidipes were significantly lower than in those in villages without that species. It is evident that the presence of G. pallidipes brings about an increase in the prevalence of T. congolense, which causes a more severe disease in cattle than other species of trypanosomes. This is a rare case of a re-invasion of a tsetse species, the disease transmission capacity of which calls for a refocusing of the traditional national tsetse and trypanosomosis control strategies to contain it.
13123 Njiokou, F., Simo, G., Nkinin, S.W., Laveissière, C. & Herder, S., 2004. Infection rate of Trypanosoma brucei s.l., T. vivax, T. congolense “forest type”, and T. simiae in small wild vertebrates in south Cameroon. Acta Tropica 92 (2): 139–146.
Njiokou: LRT, OCEAC, BP 288, Yaounde, Cameroon. [fnjiokou@ yahoo.com]
In order to identify the infection rate of trypanosome species infecting wild animals in four localities (Bipindi, Campo, Fontem and Nditam) of southern Cameroon, 1 141 wild animals were sampled. These animals belonged to 36 species grouped in 8 orders including 407 primates, 347 artiodactyls, 264 rodents, 54 pangolins, 53 small carnivores, 11 saurians and crocodilians and 5 hyraxes. PCR using specific primers for Trypanosoma vivax, T. brucei s.l., T. congolense “forest type” and T. simiae showed that 18.7 percent of the animals were infected by at least one of these trypanosome species. A positive PCR result may not indicate absolutely an active infection because PCR can detect also transient infections. T. vivax (Duttonella) had the highest infection rate (9.5 percent) and was found in almost all the host orders studied. T. brucei s.l. mostly infected primates, rodents and some duikers (Cephalophus dorsalis and C. monticola). Trypanosomes of the subgenus Nannomonas had a lower infection rate of 5.5 percent (2.4 percent for T. simiae and 3.1 percent for T. congolense “forest type”). They were harboured mainly by primates, ungulates and rodents. Trypanosome infection rates were highest in Nditam (24.5 percent) and Bipindi (21 percent). Trypanosoma brucei s.l. (Trypanozoon) had its maximum infection rate of 10.4 percent in Bipindi. The “Quantitative Buffy Coat” (QBC®) and kit for in vitro isolation (KIVI) techniques were used to identify 48 (6.1 percent) infected animals. Thirteen were positive using QBC®), and 42 were positive by KIVI. However, PCR was negative on 16 of these infected animals, probably due to infections with other trypanosome species. This study showed that trypanosomes of the subgenera Duttonella, Nannomonas and Trypanozoon could infect small wild vertebrates as has been shown for large ungulates and carnivores. The presence of T. brucei s.l. in a large range of wild animals strengthens the hypothesis of the existence of a wild animal reservoir of T. b. gambiense in Cameroon.
32 2005 Tsetse and Trypanosomiasis Information 4,5a
13124 Turner, C.M.R., McLellan, S., Lindergard, L.A.G., Bisoni, L., Tait, A. & MacLeod, A., 2004. Human infectivity trait in Trypanosoma brucei: stability, heritability and relationship to sra expression. Parasitology, 129 (4): 445–454.
Turner: Division of Infection and Immunity, Institute of Biomedical and Life Sciences, Joseph Black Building, University of Glasgow, Glasgow, G12 8QQ, UK. [[email protected]]
Some Trypanosoma brucei lines infect humans, whereas others do not because the parasites are lysed by human serum. We have developed a robust, quantitative in vitro assay based on differential uptake of fluorescent dyes by live and dead trypanosomes to quantify the extent and kinetics of killing by human serum. This method has been used to discriminate between three classes of human serum resistance: sensitive, resistant and intermediate. TREU 927/4, the parasite used for the T. brucei genome project, is intermediate. The phenotype is expressed in both bloodstream and metacyclic forms, and is stably expressed during chronic infections and on cyclical transmission through tsetse flies. Trypanosomes of intermediate phenotype are distinguished from sensitive populations of cells by the slower rate of lysis and by the potential to become fully resistant to killing by human serum as a result of selection or long-term serial passaging in mice, and to pass on full resistance phenotype to their progeny in a genetic cross. The sra gene has been shown previously to determine human serum resistance in T. brucei but screening for the presence and expression of this gene indicated that it is not responsible for the human serum resistance phenotype in the trypanosome lines that we have examined, indicating that an alternative mechanism for HSR exists in these stocks. Examination of the inheritance of the phenotype in F1 hybrids for both bloodstream and metacyclic stages from two genetic crosses demonstrated that the phenotype is co- inherited in both life-cycle stages in a manner consistent with being a Mendelian trait, determined by only one or a few genes.
5. HUMAN TRYPANOSOMIASIS
(a) SURVEILLANCE
[See also 28: 13088, 13118, 13120, 13131]
13125 Agranoff, D., Stich, A., Abel, P. & Krishna, S., 2005. Proteomic fingerprinting for the diagnosis of human African trypanosomiasis. Trends in Parasitology, 21 (4): 154–157.
Krishna: Department of Cellular and Molecular Medicine (Infectious Diseases), St George’s Hospital Medical School, Cranmer Terrace, London, SW17 ORE, UK.
33 2005 Tsetse and Trypanosomiasis Information 4,5a
Papadopoulos et al. [see TTI 27(2): 12940] recently reported the discovery of a diagnostic serum proteomic signature for human African trypanosomiasis (HAT), using a combination of surface-enhanced laser desorption-ionization time-of-flight (SELDI-TOF) mass spectrometry and data-mining algorithms. This novel approach, coupled with biochemical characterization of the proteins that contribute to the signature, provides powerful new tools for the development of improved diagnostic tests, disease staging and identification of potential novel drug targets in HAT.
13126 Anon. 2005. A new form of human trypanosomiasis in India. Weekly Epidemiological Record, 80 (7): 62–63.
Weekly Epidemiological Record: Avenue Appia 20, 1211 Geneva 27, Switzerland.
On 26th October 2004, a 40-year-old farmer working in close contact with livestock in Maharashtra State was admitted to a hospital in Nagpur, India. The patient presented with episodes of fever associated with sensory disorders. Testing of the blood revealed the presence of trypanosomes of Trypanosoma evansi. A specialist from the Institut français de recherche pour le développement (Montpellier, France) was sent by WHO to confirm the parasitological diagnosis. Treatment with suramin over three months led to a rapid improvement. At the time of this report it was assumed that the patient was cured. This is the first formally identified human case in the world of trypanosomiasis caused by T. evansi.
13127 Hutchinson, O.C., Webb, H., Picozzi, K., Welburn, S. & Carrington, M., 2004. Candidate protein selection for diagnostic markers of African trypanosomiasis. Trends in Parasitology, 20 (11): 519–523.
Carrington: Institute of Zoology, Zoological Society of London, Regent's Park, London, NW1 4RY, UK.
The ability to accurately diagnose the presence of an infective microorganism is not only important for individual human and animal health and wellbeing, but is also central to surveillance programmes. Effective and sustainable control of many diseases in the developing world depends on the availability of field applicable diagnostics that are cheap, reliable, simple in design and application, and which provide immediate results. This review examines how the genome sequences can be used in the selection of potential candidate proteins for developing new serodiagnostics for African trypanosomiasis.
13128 Ngoma, M.S., Nalubamba, M., Kowa, S., Minyoi, D., Mubanga, J., Mwansa, J., Musuku, J., Chintu, C., Bhat, G., Kapakala, T., Mumba, P. & Nyalungwe, K., 2004. Congenital trypanosomiasis. Journal of Tropical Pediatrics, 50 (6): 377–381.
Ngoma: School of Medicine, Department of Paediatrics and Child Health, PO Box 50110, Lusaka, Zambia. [[email protected]]
34 2005 Tsetse and Trypanosomiasis Information 5a,b
The last successfully treated case of congenital trypanosomiasis in Zambia was in October 1978, with detailed analysis of immunoglobulins, illustrating the waning of blood and serum levels of IgA, IgG, and IgM during treatment, up to 99 days after treatment. Twenty-five years later, we report on a case of congenital trypanosomiasis. The disease is now rare and can be missed or dismissed as retroviral disease, particularly in adults. The main unusual symptoms were the prolonged intermittent convulsions in an otherwise well infant. Management of the disease is now more interdisciplinary, resources for laboratory support are fewer, lumbar puncture is more relevant, and antitrypanosomal drugs are more difficult to obtain. The mother died within one week of hospitalization and the infant initially responded to three doses of suramin and 3 weeks of melsopropol. Convulsions ceased during the second round of melsopropol. Unfortunately, the infant died of nosocomial infection.
(b) PATHOLOGY AND IMMUNOLOGY
[See also 28: 13088, 13124]
13129 Buguet, A., Bisser, S., Josenando, T., Chapotot, F. & Cespuglio, R., 2005. Sleep structure: a new diagnostic tool for stage determination in sleeping sickness. Acta Tropica, 93 (1): 107–117.
Buguet: EA 3734 Neurobiologie des états de Vigilance, IFR-19, Claude- Bernard-Lyon 1 University, 8 Avenue Rockefeller, 69373 Lyon Cedex 08, France.
Human African trypanosomiasis (HAT), due to the transmission of Trypanosoma brucei gambiense and T. b. rhodesiense by tsetse flies, is re-emerging in intertropical Africa. It evolves from the hemolymphatic Stage I to the meningo-encephalitic Stage II. The latter is generally treated with melarsoprol, an arseniate often provoking a deadly encephalopathy. A precise determination of the HAT evolution stage is therefore crucial. Stage II patients show: (i) a deregulation of the 24-h distribution of the sleep-wake alternation; (ii) an alteration of the sleep structure, with frequent sleep onset rapid eye movement (REM) periods (SOREMPs). Gambian HAT was diagnosed in eight patients (four, Stage II; three, Stage I; one, “intermediate” case) at the trypanosomiasis clinic at Viana (Angola). Continuous 48-h polysomnography was recorded on Oxford Medilog 9000-II portable systems before and after treatment with melarsoprol (Stage II) or pentamidine (Stage I and “intermediate” stage). Sleep traces were visually analyzed in 20-s epochs using the PRANA software. Stage II patients showed the complete sleep- wake syndrome, partly reversed by melarsoprol 1 month later. Two Stage I patients did not experience any of these alterations. However, the “intermediate” and one Stage I patients exhibited sleep disruptions and/or SOREMPs, persistent after pentamidine treatment. Polysomnography may represent a diagnostic tool to distinguish the two stages of HAT. Especially, SOREMPs appear shortly after the central nervous system invasion
35 2005 Tsetse and Trypanosomiasis Information 5a,b by trypanosomes. The reversibility of the sleep-wake cycle and sleep structure alterations after appropriate treatment constitutes the basis of an evaluation of the healing process.
13130 Girard, M., Giraud, S., Courtioux, B., Jauberteau-Marchan, M.-O. & Bouteille, B., 2004. Endothelial cell activation in the presence of African trypanosomes. [human bone marrow endothelial cell] Molecular and Biochemical Parasitology, 139 (1): 41–49.
Girard: EA 3174 Neuroparasitologie et Neuroépidémiologie Tropicales, Faculté de medecine, Limoges, France.
During human African trypanosomiasis, trypanosomes (Trypanosoma brucei gambiense or T. b. rhodesiense) invade the central nervous system (CNS). Mechanisms of blood-brain barrier and blood-cerebrospinal fluid barrier leakage remain unknown. To better understand the relationships between trypanosomes and endothelial cells, the principal cell population of those barriers, we cultured a human bone marrow endothelial cell (HBMEC) line in the presence or absence of T. b. gambiense, to study cell activation. As indicated by NF-țB translocation to the nucleus, cells were activated in the presence of trypanosomes. The expression of the adhesion molecules ICAM-1, E-selectin and VCAM-1 increased in co-culture. The parasites induced the synthesis of the pro- inflammatory cytokines TNF-Į, IL-6 and IL-8, and of nitric oxide (NO) by HBMEC. Cells were also cultured in the presence of parasite variant surface glycoproteins (VSGs), and an increase in TNF-Į, IL-6, IL-8, and NO synthesis was also observed. Soluble VSGs induced NF-țB translocation, and the expression of adhesion molecules, indicating that they could possibly be the molecular soluble factor responsible for endothelial cell activation. The permeability coefficient of the HBMEC layer increased when cells were cultured in the presence of trypanosomes, parasite culture supernatant, or VSGs. Thus, T. b. gambiense can activate endothelial cells in vitro, through the release of soluble activating factors. Consequences of endothelial cell activation by parasite products may include a potentiation of the inflammatory reaction, leukocyte recruitment, passage of trypanosomes into the CNS, and barrier dysfunction observed during CNS involvement of HAT.
13131 Lejon, V. & Büscher, P., 2005. Cerebrospinal fluid in human African trypanosomiasis: a key to diagnosis, therapeutic decision and post-treatment follow-up. [Review] Tropical Medicine and International Health, 10 (5): 395– 403.
Lejon: Department of Parasitology, Institute of Tropical Medicine, Nationalestraat 155, 2000 Antwerpen, Belgium. [[email protected]]
Human African trypanosomiasis is a lethal parasitic infection with neurological involvement. Examination of the cerebrospinal fluid (CSF) plays an essential role in diagnosis, selection of treatment and post-treatment follow-up. This paper reviews clinical presentation, diagnosis and treatment of the disease, with emphasis on CSF
36 2005 Tsetse and Trypanosomiasis Information 5b characteristics and interpretation of the CSF results for therapeutic decision and post- treatment follow-up.
13132 Lugli, E.B., Pouliot, M., Portela, M. del P.M., Loomis, M.R. & Raper, J., 2004. Characterization of primate trypanosome lytic factors. Molecular and Biochemical Parasitology, 138 (1): 9–20.
Raper: Department of Medical and Molecular Parasitology, New York University School of Medicine, 341, East 25th Street, New York, NY 10010, USA.
Humans are one of the few species that resist infection by Trypanosoma brucei brucei because the parasites are killed by lytic factors found in human serum. Trypanosome lytic factors (TLFs) are protein/lipid complexes that contain apolipoprotein A-I (apoA-I), and are therefore a class of high density lipoproteins (HDLs). Haptoglobin- related protein (Hpr) is a unique protein component of TLFs, and its expression has only been demonstrated in humans. Trypanolytic activity has only been found in the sera of five primates: humans, gorillas, mandrills, baboons and sooty mangabeys. We describe here previously unidentified components of highly purified human TLF1: apolipoprotein L-I (apoL-I), human cathelicidin antimicrobial peptide 18 (hCAP18) and glycosyl- phosphatidylinositol-specific phospholipase D (GPI-PLD). However, we found that hCAP18 and GPI-PLD, along with apoA-I, are common components of both lytic and non-lytic primate HDLs. In contrast, Hpr, which has been previously implicated as the main lytic component of TLF1, was a unique component of all trypanolytic primate HDLs. Furthermore, a polyclonal antiserum to Hpr neutralised the lytic activity from humans and baboons. ApoL-I, a candidate lytic component of human serum, was not immunologically or genetically detectable in two primate species with lytic activity. Polyclonal antiserum to apoL-I also did not neutralise TLF activity in a total human HDL preparation. These findings suggest that apoL-I is not essential in all primate TLFs, and apoL-I alone is not sufficient for optimal trypanosome lytic activity in human TLF.
13133 Lundkvist, G.B., Kristensson, K. & Bentivoglio, M., 2004. Why trypanosomes cause sleeping sickness. Physiology, 19 (4): 198–206.
Kristensson: Department of Neuroscience, Karolinska Insitutet, Stockholm, SE-171 77 Sweden. [[email protected]]
African trypanosomiasis or sleeping sickness is hallmarked by sleep and wakefulness disturbances. In contrast to other infections, there is no hypersomnia, but the sleep pattern is fragmented. This overview discusses how the causative agents, the parasites Trypanosoma brucei, target circumventricular organs in the brain, causing inflammatory responses in hypothalamic structures that may lead to dysfunctions in the circadian-timing and sleep-regulatory systems.
13134 Lutumba, P., Robays, J., Miaka, C., Kande, V., Simarro, P.P., Shaw, A.P.M., Dujardin, B. & Boelaert, M., 2005. Efficience de différentes stratégies de
37 2005 Tsetse and Trypanosomiasis Information 5b
détection de la trypanosomiase humaine Africaine à T. b. gambiense [Efficiency of different detection strategies concerning human African trypanosomiasis due to T. b. gambiense.] Tropical Medicine and International Health, 10 (4): 347–356.
Boelaert: Unité d’épidémiologie, Institut de Médecine Tropicale Prince Léopold, Nationalestraat 155, 2000 Anvers, Belgique. [mboelaert@itg. be]
Population screening for human African trypanosomiasis (HAT) is often based on a combination of two screening tests: lymph node palpation (LN) and card agglutination test for trypanosomiasis (CATT). This decision analysis compared the efficiency of three alternative detection strategies: screening by LN only, CATT only and their combination (LN and CATT). An HAT detection strategy was defined as the sequence of screening and confirmation. Efficacy was evaluated in terms of lives saved. The cost of screening and confirmation tests was estimated in US$. The different parameters in the decision tree were based on published literature and observations of the HAT control programme in the Democratic Republic of Congo. A sensitivity analysis was carried out on those parameters subject to uncertainty. The cost-effectiveness of a detection strategy based on CATT was US$125 per life saved, compared with US$517 for LN and US$452 for the combined. Marginal cost to add LN to CATT only was between US$1 225 and US$5 000 per life saved. Sensitivity analysis shows that these results are robust to variation. The CATT strategy was the most efficient. None of the strategies was able to avoid more than 60 percent of HAT deaths. This moderate efficacy is due to the low sensitivity of the confirmatory (diagnostic) tests. Substantial efficiency gains can be obtained by adopting a CATT only strategy and resources can be better allocated to more sensitive confirmatory tests or to increasing the coverage of populations at risk.
13135 Monteiro, R.Q., Campana, P.T., Melo, P.A. & Bianconi, M.L., 2004. Suramin interaction with human Į-thrombin: inhibitory effects and binding studies. International Journal of Biochemistry & Cell Biology, 36 (10): 2077–2085.
Monteiro: Departamento de Bioquímica Médica, ICB/CCS, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil. [[email protected]]
Suramin is a hexasulfonated naphthylurea commonly used as antitrypanosomal drug and more recently for the treatment of malignant tumours. Here we show that suramin binds to human Į-thrombin inhibiting both the hydrolysis of the synthetic substrate S-2238 (IC50=40 μM), and the thrombin-induced fibrinogen clotting (IC50=20 μM). The latter is completely reversed by albumin (30 mg mL-1) suggesting that, at therapeutic concentrations, suramin is unable to affect Į-thrombin activity in the plasma. Kinetic analysis showed that suramin acts as a non-competitive inhibitor decreasing Vmax without changing the Km for S-2238 hydrolysis. Calorimetric studies revealed two distinct binding sites for suramin in Į-thrombin. In addition, circular dichroism studies showed that suramin causes significant changes in Į-thrombin tertiary structure, without affecting the secondary structure content. Interaction with Į-thrombin resulted in an increased
38 2005 Tsetse and Trypanosomiasis Information 5b fluorescence emission of the drug. Complex formation was strongly affected by high ionic strength suggesting the involvement of electrostatic interactions. Altogether our data suggest that part of the biological activities of suramin might be related to Į- thrombin inhibition at extra-vascular sites.
13136 Rock, R.B., Gekker, G., Hu, S.X., Sheng, W.S., Cheeran, M., Lokensgard, J.R. & Peterson, P. K., 2004. Role of microglia in central nervous system infections. Clinical Microbiology Reviews, 17 (4): 942–964.
Peterson: Neuroimmunology Laboratory, Minneapolis Medical Research Foundation, and University of Minnesota Medical School, Minneapolis, Minnesota Department of Medicine, Hennepin County Medical Center, 701 Park Ave., Minneapolis, MN 55415 USA. [[email protected].]
The nature of microglia fascinated many prominent researchers in the 19th and early 20th centuries, and in a classic treatise in 1932, Pio del Rio-Hortega formulated a number of concepts regarding the function of these resident macrophages of the brain parenchyma that remain relevant to this day. However, a renaissance of interest in microglia occurred toward the end of the 20th century, fuelled by the recognition of their role in neuropathogenesis of infectious agents, such as human immunodeficiency virus type 1, and by what appears to be their participation in other neurodegenerative and neuroinflammatory disorders. During the same period, insights into the physiological and pathological properties of microglia were gained from in vivo and in vitro studies of neurotropic viruses, bacteria, fungi, parasites, and prions, which are reviewed in this article. New concepts that have emerged from these studies include the importance of cytokines and chemokines produced by activated microglia in neurodegenerative and neuroprotective processes and the elegant but astonishingly complex interactions between microglia, astrocytes, lymphocytes, and neurons that underlie these processes. It is proposed that an enhanced understanding of microglia will yield improved therapies of central nervous system infections, since such therapies are, by and large, sorely needed.
13137 Truc, P., 2003. About Trypanosoma brucei gambiense, the causative agent of the chronic form of Human African Trypanosomiasis: some findings and proposals. African Journal of Biotechnology, 2 (12): 657–661
Truc: Organisation de Coordination pour la lutte contre les Endémies en Afrique Centrale (OCEAC), Department of Research and Control of Human African Trypanosomiasis, BP 288, Yaounde, Cameroon. [[email protected]]
Since its discovery at the beginning of 20th century, Human African Trypanosomiasis or sleeping sickness has killed several hundred thousands of individuals in Africa, mainly in the central part of the continent. The present status of the disease in several countries is dramatic, such as the Sudan, Democratic Republic of Congo or Angola. However, diagnosis is still based on the detection of parasites in blood or lymphatic juice by microscopy, as in the 1930s. Furthermore, since the discovery of melarsoprol in 1949, no new drugs for treatment have been synthesized. We have to see
39 2005 Tsetse and Trypanosomiasis Information 5b
HAT as coming under the head of ‘most-neglected diseases’ but some findings obtained through biotechnology are discussed in terms of epidemiology and control applications.
(c) TREATMENT
[See also 28: 13087, 13088, 13090, 13129]
13138 Deterding, A., Dungey, F.A., Thompson, K.A. & Steverding, D., 2005. Anti- trypanosomal activities of DNA topoisomerase inhibitors. Acta Tropica, 93 (3): 311–316.
Steverding: School of Medicine, Health Policy and Practice, University of East Anglia, Norwich NR4 7TJ, UK.
Only four drugs are available for chemotherapy of human African sleeping sickness and these have undesirable toxic side effects. The development of new anti-trypanosomal drugs is, therefore, urgently required. In this study, 15 DNA topoisomerase inhibitors, including approved anti-cancer drugs, were tested for in vitro activity against bloodstream forms of Trypanosoma brucei and human leukaemia HL-60 cells. All compounds exhibited anti-trypanosomal activity, with ED50 values ranging between 3 nM and 30 ȝM, and MIC values between 100 nM and >100 ȝM. The trypanocidal activities of the most effective DNA topoisomerase inhibitors, aclarubicin, doxorubicin and mitoxantrone, were comparable with those of commercial anti-trypanosomal drugs. These data support the use of DNA topoisomerase inhibitors as lead compounds for anti- trypanosomal drug development.
13139 Miezan T.W., Dje N.N., Doua F. & Boa F., 2002. Trypanosomose humaine africaine en Côte d’Ivoire: caractéristiques biologiques après traitement. A propos de 812 cas traités dans le foyer de Daloa (Côte d’Ivoire). [Human trypanosomiasis in Côte d’Ivoire (Ivory Coast): biological characteristics after treatment. A study of 812 cases treated in the Daloa focus (Côte d’Ivoire).] Bulletin de la Société de Pathologie exotique, 95 (5): 362–365.
Miezan: Projet de recherches cliniques sur la trypanosomiase, BP 1425 Daloa, Côte d’Ivoire.
The treatment and post-therapeutic follow up of patients diagnosed with HAT are important for HAT control. A longitudinal survey was implemented in the focus of Daloa (Côte d’Ivoire). A total of 812 patients infected with Trypanosoma brucei gambiense in meningoencephalitic stage and treated with melarsoprol were included; this study focussed on the biological characteristics of patients after treatment. The relapse occurs between 1 and 24 months after treatment. It is essentially neurological, and characterised by the presence in the CSF of antibodies, by the increase of cell count compared with value immediately after treatment, or by the presence of trypanosomes. The cure can be confirmed from 18 months after treatment, and is characterised by the absence of
40 2005 Tsetse and Trypanosomiasis Information 5c antibodies and trypanosomes in the CSF, by a normal cell count and a normal proteinorachy. However, some biological damage was noted in some patients by the end of the 18-month follow-up period but no relapses occurred amongst them.
13140 Moore, A.C., 2005. Prospects for improving African trypanosomiasis chemotherapy. Journal of Infectious Diseases, 191 (11): 1793–1795.
Moore: Division of Parasitic Diseases, M/S F-22, Centers for Disease Control and Prevention, 4770 Buford Hwy., Atlanta, GA 30341 USA. [aym2@ cdc.gov].
Nearly a century ago, specific antimicrobial agents for the treatment of human African trypanosomiasis (HAT) were among the first achievements in the new science of drug discovery and development. However, few advances in HAT chemotherapy have been made in recent decades, and patients today are usually treated with drugs that were introduced before 1950. The infection is transmitted only in rural Africa and does not have a profitable market that would encourage drug development. This lack of progress is part of a larger trend in the global pharmaceutical industry. Investment in treatment for tropical parasitic diseases has plummeted, despite the increasing health burden of these infections. Less than 1 percent of the new chemical entities introduced during the mid-1970s through 1999 were developed for tropical disease indications, and few candidate drugs are in development. Insufficient financial return has not only discouraged research and development but also caused companies to withdraw existing antiparasitic drugs from the market. Five years ago, when virtually every drug capable of curing central nervous system (CNS) infection was no longer produced or was threatened, it appeared that HAT might become untreatable. At present, these drugs are back in production and are available from the World Health Organization (WHO) through donation programs, but a method to ensure availability for the long term has not been identified. The current serious resurgence of the disease and the present inadequacy of the available drugs to treat it are described. Trials of new drugs and improved methods of applying the existing drugs give hope for some improvement in the present serious situation.
13141 Pépin, J. & Mpia, B., 2005. Trypanosomiasis relapse after melarsoprol therapy, Democratic Republic of Congo, 1982-2001. Emerging Infectious Diseases, 11 (6): 921–927.
Pépin: Centre for International Health, 3001, 12ème Avenue Nord, Sherbrooke, Québec J1H 5N4, Canada. [[email protected]]
Recently, a high proportion of patients with late-stage Trypanosoma brucei gambiense trypanosomiasis, that had been treated with melarsoprol in some disease- endemic areas, subsequently relapsed. To determine whether the frequency of post- melarsoprol relapses increased over time, we reviewed data from 2 221 trypanosomiasis patients treated with melarsoprol during this period in Nioki, Democratic Republic of Congo, from 1982 to 2001. The frequency of relapses was 5.6 percent (31/553), 6.8 percent (35/512), 4.5 percent (18/398), 11.4 percent (34/299), and 5.0 percent (17/343)
41 2005 Tsetse and Trypanosomiasis Information 5c for those treated from 1982 to 1985, 1986 to 1989, 1990 to 1993, 1994 to 1997, and 1998 to 2001, respectively. The higher frequency of relapses in 1994 to 1997 was associated with an incremental dosage regimen of melarsoprol. In multivariate analysis, after adjustment for treatment regimen, sex, residence, and trypanosomes in cerebrospinal fluid, post-melarsoprol relapses were shown not to have increased in Nioki, perhaps because 1) little drug pressure exists; 2) sub-therapeutic doses have rarely been administered; 3) little potential exists for the preferential transmission of melarsoprol- resistant strains.
13142 Sanchez-Delgado, R.A. & Anzellotti, A., 2004. Metal complexes as chemotherapeutic agents against tropical diseases: Trypanosomiasis, malaria and leishmaniasis. Mini-Reviews in Medicinal Chemistry, 4 (1): 23–30.
Sanchez-Delgado: Instituto Venezolano de Investigaciones Científicas (IVIC), Apartado 21827 Caracas 1020-A, Venezuela.
Parasitic diseases represent a major world health problem with very limited therapeutic options, most of the available treatments being decades old and suffering from limited efficacy and/or undesirable collateral effects. The use of metal complexes as chemotherapeutic agents against these ailments appears as a very attractive alternative. Although the design of metal complexes with good therapeutic index is still rather empirical, a number of potential metal-based antiparasitic drugs have become available. In this review, advances in the use of metal complexes for the treatment of trypanosomiasis, malaria, and leishmaniasis as important representatives of the general area of tropical diseases are described.
13143 Schmid, C., Nkunku, S., Merolle, A., Vounatsou, P. & Burri, C., 2004. Efficacy of 10-day melarsoprol schedule 2 years after treatment for late-stage gambiense sleeping sickness. Lancet, 364 (9436): 789–790.
Burri: Swiss Tropical Institute, 4002 Basel, Switzerland. [[email protected]]
In 2000, we reported that a new short treatment schedule of melarsoprol was not worse than the longer and demanding standard treatment for late-stage human African trypanosomiasis. This alternative schedule was assessed in an open, randomised clinical equivalence trial of 500 patients in Angola. At 24h after treatment, all patients were parasite free. Of 442 patients, 12 (3 percent) had relapsed after one year, of whom seven (3 percent) had had standard treatment and five (2 percent) the alternative treatment. After two years, 23 (5 percent) relapsing patients were reported, 11 (5 percent) in the standard treatment group and 12 (6 percent) in the new group. The results at the two-year follow-up support and strengthen our previous findings.
13144 Schmid, C., Richer, M., Bilenge, C.M.M., Josenando, T., Chappuis, F., Manthelot, C.R., Nangouma, A., Doua, F., Asumu, P.N., Simarro, P.P. & Burri, C., 2005. Effectiveness of a 10-day melarsoprol schedule for the
42 2005 Tsetse and Trypanosomiasis Information 5c
treatment of late-stage human African trypanosomiasis: Confirmation from a multinational study (IMPAMEL II). Journal of Infectious Diseases, 191 (11): 1922–1931.
Burri: Swiss Tropical Institute, Socinstrasse 57, CH-4002 Basel, Switzerland. [[email protected]]
Treatment of late-stage human African trypanosomiasis (HAT) with melarsoprol can be improved by shortening the regimen. A previous trial demonstrated the safety and efficacy of a 10-day treatment schedule. We demonstrate the effectiveness of this schedule in a non-controlled, multinational drug-utilization study. A total of 2 020 patients with late-stage HAT were treated with the 10-day melarsoprol schedule in 16 centres in seven African countries. We assessed outcome on the basis of major adverse events and the cure rate after treatment and during two years of follow-up. The cure rate 24 h after treatment was 93.9 percent; two years later, it was 86.2 percent. However, 49.3 percent of patients were lost to follow-up. The overall fatality rate was 5.9 percent. Of treated patients, 8.7 percent had an encephalopathic syndrome that was fatal 45.5 percent of the time. The rate of severe bullous and maculopapular eruptions was 0.8 percent and 6.8 percent, respectively. The 10-day treatment schedule was well implemented in the field and was effective. It reduces treatment duration, drug amount, and hospitalization costs per patient, and it increases treatment-centre capacity. The shorter protocol has been recommended by the International Scientific Council for Trypanosomiasis Research and Control for the treatment of late-stage HAT caused by Trypanosoma brucei gambiense.
13145 Sternberg, J.M., 2004. Human African trypanosomiasis: clinical presentation and immune response. Parasite Immunology, 26 (11–12): 469–476.
Sternberg: School of Biological Sciences, University of Aberdeen, Aberdeen AB24 2TZ, UK.
Human African trypanosomiasis or sleeping sickness is caused by infection with two subspecies of the tsetse-fly-vectored haemoflagellate parasite Trypanosoma brucei. Historically, epidemic sleeping sickness has caused massive loss of life, and related animal diseases have had a crucial impact on development in sub-Saharan Africa. After a period of moderately successful control during the mid-part of the 20th century, sleeping sickness incidence is currently rising, and control is hampered by a combination of factors, including civil unrest and the possible development of drug resistance by the parasites. The prevailing view is that the disease is invariably fatal without anti- trypanosomal drug treatment. However, there have also been intriguing reports of wide variations in disease severity as well as evidence of asymptomatic carriers of trypanosomes. These differences in the presentation of the disease will be discussed in the context of our knowledge of the immunology of trypanosomiasis. The impact of dysregulated inflammatory responses in both systemic and CNS pathology will be examined and the potential for host genotype variation in disease severity and control will be discussed.
43 2005 Tsetse and Trypanosomiasis Information 5c
6. ANIMAL TRYPANOSOMIASIS
(a) SURVEY AND DISTRIBUTION
13146 Abenga, J.N., David, K., Ezebuiro, C.O.G., & Lawani, F.A.G., 2005. Observations on the tolerance of young dogs (puppies) to infection with Trypanosoma congolense. African Journal of Clinical and Experimental Microbiology, 6 (1): 28–33.
Abenga: Nigerian Institute for Trypanosomiasis Research, PMB 2077, Kaduna, Nigeria. [[email protected]]
Studies were undertaken to assess the susceptibility of young local dogs to infection with Trypanosoma congolense. Six puppies (7 weeks old) were used for the study. Although the puppies became parasitaemic 6 to 7 days post infection, they were tolerant to infection as the parasitaemia remained low throughout the first seven weeks of the eight-week observation period. The packed cell volume also only dropped slightly during the last four weeks while the mean body weight continued to increase. Similarly, the mean daily body temperature did not differ significantly from those of uninfected control. The significance of trypanotolerance in Nigerian local dogs is discussed.
13147 Abenga, J.N., Enwezor, F.N.C., Lawani, F.A.G., Ezebuiro, C., Sule, J. & David, K.M., 2002. Prevalence of trypanosomosis in trade cattle at slaughter in Kaduna, Nigeria. Nigerian Journal of Parasitology, 23: 107–110.
Abenga: Pathology, Epidemiology and Statistics Division, Nigerian Institute for Trypanosomiasis Research, PMB 2077, Kaduna, Kaduna State, Nigeria. [[email protected]]
The prevalence of trypanosomosis in trade cattle at slaughter in Kaduna was investigated during the peak of the rainy season (July to September, 1999). A total of 500 blood samples was collected and parasitologically examined using the haematocrit centrifugation technique and buffy coat method. Physical examination of the animals was done before and after slaughter to assess their clinical condition. The overall infection rate was 3.8 percent. However, 30.9 percent of the animals showed severe emaciation. The infection rate in emaciated animals (7.6 percent) was higher than in non-emaciated animals (2.1 percent). Infected cattle also had significantly lower packed cell volume values than non-infected cattle. The prevalence of trypanosomosis observed in this study does not reflect the true status of the disease in Kaduna State. However, these findings indicate that trypanosomosis has an impact on the physical condition of cattle at slaughter and consequently on the market value.
13148 Abenga, J.N., Enwezor, F.N.C., Lawani, F.A.G., Osue, H.O. & Ikemereh, E.C.D., 2004. Trypanosome prevalence in cattle in Lere area in Kaduna State,
44 2005 Tsetse and Trypanosomiasis Information 6a
North Central Nigeria. Revue d'Élevage et de Médecine vétérinaire des Pays tropicaux, 57 (1–2): 45–48.
Abenga: Pathology, Epidemiology and Statistics Division, Nigerian Institute for Trypanosomiasis Research, PMB 2077, Kaduna, Kaduna State, Nigeria. [[email protected]]
Trypanosome prevalence in cattle was estimated in August 2001 in selected farms in the three districts of Lere local government area of Kaduna State, North Central Nigeria. The study was subsequent to reports of trypanosomosis outbreaks, which had resulted in deaths of cattle that led to early migration of semi-nomadic Fulanis out of the area, especially during the rains. Blood samples collected randomly from 526 cattle were examined for presence of trypanosomes using the buffy coat technique and Giemsa thin blood smears. Overall, 48 animals were found infected: 39 (81 percent) with Trypanosoma vivax, 7 (15 percent) with T. congolense and 2 (4 percent) with T. brucei. The infection rates in young and adult cattle were 6 and 10 percent, respectively, which were not significantly different. From this study, it is clear that trypanosomosis is still a major obstacle to livestock production in Nigeria and that the incidence rate is similar in young and adult animals.
13149 Cherenet, T., Sani, R.A., Panandam, J.M., Nadzr, S., Speybroeck, N. & van den Bossche, P., 2004. Seasonal prevalence of bovine trypanosomosis in a tsetse-infested zone and a tsetse-free zone of the Amhara Region, north-west Ethiopia. Onderstepoort Journal of Veterinary Research, 71 (4): 307–312.
Cherenet: Faculty of Veterinary Medicine, University Putra Malaysia, 43400 Serdang, Malaysia.
During a period of four consecutive years, trypanosomosis surveys were conducted in a tsetse-infested and tsetse-free area of the Amhara Region of north-west Ethiopia. In each study area randomly selected communal cattle were sampled and their blood was investigated using parasitological diagnostic methods. At the same time the population of biting flies was sampled. The monthly average prevalence of trypanosome infections in cattle did not differ significantly between study areas. In both study areas, the prevalence of trypanosome infections was highest during the long rainy season. Trypanosome infections were mainly due to Trypanosoma vivax and they significantly reduced the average packed cell volume and the body condition of the animals. The monthly prevalence of infection was correlated with the density of biting flies, such as Tabanidae and Stomoxys spp., in the preceding month suggesting an important role of mechanical transmission in the epidemiology of trypanosomosis in both areas.
13150 Dinka, H. & Abebe, G., 2005. Small ruminants trypanosomosis in the southwest of Ethiopia. Small Ruminant Research, 57 (2–3): 239–243.
45 2005 Tsetse and Trypanosomiasis Information 6a
Abebe: Department of Parasitology, Faculty of Veterinary Medicine, Addis Ababa University, P.O. Box 34, Debre Zeit, Ethiopia.
A study was conducted in two valleys of the southwest Ethiopia (Didessa and Ghibe valleys) from November 2002 to April 2003 to collect baseline data on the prevalence of trypanosomosis in local breeds of sheep and goats. Blood samples from 533 randomly selected small ruminants of different species, sex and age groups were collected and examined with conventional haematological and parasitological techniques. Among the small ruminants examined during the study period 27 animals (5.1 percent) were infected with trypanosomes. Most of the infections were due to Trypanosoma congolense (46.7 percent, 33.3 percent) followed by T. vivax (26.7 percent, 25.0 percent) and the rest was due to T. brucei (6.7 percent, 8.3 percent) and mixed infections of T. congolense and T. vivax (13.3 percent, 25.0 percent), T. brucei and T. vivax (6.7 percent, 8.3 percent) in sheep and goats, respectively. There was no statistically significant difference in infection between male and female, among age groups in sheep and goats as well as valleys. Infection between sheep and goats showed significant difference, the higher being 7.7 percent in sheep and the lower 3.6 percent in goats. Mean packed cell volume (PCV) value of parasitaemic animals was significantly lower than that of aparasitaemic animals. In an attempt to identify the vectors involved in the transmission of small ruminants trypanosomosis, both tsetse flies of the morsitans group (Glossina pallidipes and G. morsitans submorsitans) and palpalis group (G. fuscipes fuscipes) and mechanical vectors of trypanosomosis that belong to Tabanidae (Tabanus) were captured in the lowlands of Didessa (1400–1780 m above sea level) and Ghibe (1250–1700 m above sea level) valleys. The study revealed that trypanosomosis in sheep and goats is an important disease and small ruminants serve as a potential reservoir of infection for other animals.
13151 Magona, J.W., Walubengo, J. & Olaho-Mukani, W., 2004. Knowledge and attitudes of cattle owners regarding trypanosomosis control in tsetse-infested areas of Uganda. Journal of the South African Veterinary Association, 75 (4): 173–176.
Magona: Ministry of Agricultural and Animal Industries & Fisheries, Animal Resources, POBox 105, Entebbe, Uganda.
A pilot survey using a structured questionnaire was conducted in Tororo and Busia districts of Uganda on the knowledge and attitudes of cattle owners regarding tsetse fly and trypanosomosis control, in order to understand factors that hindered their full participation. A total of 81 cattle owners were randomly selected and interviewed, of which 92.5 percent were aware of tsetse flies and trypanosomosis and 87.6 percent recognised animal trypanosomosis as a problem in the area. Most cattle owners were aware of tsetse fly trapping (76.5 percent), isometamidium chloride use (55.5 percent), diminazene aceturate use (48 percent) and pour-on applications (18.5 percent). However, knowledge did not coincide with the application of control measures. Despite the widespread awareness, tsetse fly trapping and pour-on applications were used by only a small percentage of cattle owners (7.5 percent applied tsetse fly trapping while 76.5
46 2005 Tsetse and Trypanosomiasis Information 6a percent were aware of it; 1.2 percent applied pour-on insecticides while 18.5 percent were aware of them). Differences between awareness and application were highly significant for tsetse fly trapping and pour-on applications, but not for isometamidium chloride use and diminazene aceturate use. Most cattle owners (97.5 percent) were willing to participate in future control programmes, but preferred participating on a group basis (85.2 percent) rather than individually (14.8 percent). The four most favoured control options in order of importance were: fly traps supplied by the government and maintained by cattle owners; contribution of labour by cattle owners for trap deployment; self- financing of trypanocidal drugs and self-financing of pour-on insecticide. The control options that should be selected in order to elicit full participation by cattle owners are discussed.
13152 Mahama, C.I., Desquesnes, M., Dia, M.L., Losson, B., De Deken, R., Speybroeck, N. & Geerts, S., 2005. A longitudinal epidemiological survey of bovine trypanosomosis and its vectors in the White Volta river basin of Northern Ghana. Veterinary Parasitology, 128 (3–4): 201–208.
Geerts: Animal Health Department, Institute of Tropical Medicine, Nationalestraat 155, B-2000, Antwerp, Belgium.
A longitudinal epidemiological survey of bovine trypanosomosis and its vectors was carried out in the Volta river basin of Northern Ghana to determine the relationship between cattle management and the incidence of bovine trypanosomosis. Two groups of sentinel cattle under different systems of management, classified as “fully-sedentary” and “partially-sedentary”, were followed over a 1-year period starting from March 2003 onwards. Cattle were screened at intervals of three months using the buffy coat technique (BCT). Buffy coat specimen from animals that were positive for the BCT and those that were negative, but with a packed cell volume (PCV) of less than 21 percent, were further tested using the polymerase chain reaction (PCR). Plasma from all animals was tested for antibody using the indirect antibody enzyme-linked immunosorbent assay (ELISA). Trypanosomosis challenge was determined in tandem with the epidemiological survey with watering sites of sentinel cattle being the foci of interest. The parasitological prevalence at the start of the survey was higher in the fully-sedentary group (9 percent) than in the partially-sedentary group (3 percent). In subsequent visits, however, the parasitological incidence was consistently higher in the partially-sedentary group than in the fully-sedentary group. The mean seroprevalence (ELISA) of both groups increased from 3 percent in March to 54 percent in December. Statistical analysis of the serological results using a random effect logistic regression, showed a significant difference in incidence of bovine trypanosomosis between the two groups. There was also a significant effect of time. The influence of cattle herding on host-vector-parasite interface and its consequence on the incidence of trypanosomosis are discussed.
13153 Mochabo, K.O.M., Kitala, P.M., Gathura, P.B., Ogara, W.O., Catley, A., Eregae, E.M. & Kaitho, T.D., 2005. Community perceptions of important
47 2005 Tsetse and Trypanosomiasis Information 6a
camel diseases in Lapur Division of Turkana District, Kenya. Tropical Animal Health and Production, 37 (3): 187–204.
Mochabo: Kenya Trypanosomiasis Research Institute (KETRI), PO Box 362, Kikuyu, Kenya.
This paper presents the results of a study conducted in Lapur Division of Turkana District, Kenya, to estimate the incidence and mortality of camel trypanosomosis using participatory methods. Four livestock camps (‘adakars’) were selected for the study. Four informant groups comprising 6–8 key persons were used for the participatory exercises. The camel diseases identified by the pastoralists in their order of importance according to annual incidence were: trypanosomosis (11.4 percent); mange (10.8 percent); tick infestation (7.9 percent); haemorrhagic septicaemia (7.7 percent); and non-specific diarrhoea (7.6 percent). Almost half (49.3 percent) of the camel population suffered from at least one disease over the previous year. The annual incidence and mortality rates of trypanosomosis were estimated at 15 percent and 9.9 percent in adult camels and 6.9 percent and 5.2 percent in young camels, respectively. There was a seasonal occurrence of trypanosomosis, with most cases reported in the dry season. The prevalence levels of the disease reportedly declined from about 100 percent in 1978 to an almost stable state of about 15 percent in 2002. This study revealed that camel trypanosomosis is still an important disease in Turkana District, exacting a heavy toll in terms of morbidity and mortality. The economic losses due to the disease were likely to have been great owing to the central role the camel plays in this arid district of Kenya.
13154 Njiru, Z.K., Constantine, C.C., Guya, S., Crowther, J., Kiragu, J.M., Thompson, R.C.A. & Dávila, A.M.R., 2005. The use of ITS1 rDNA PCR in detecting pathogenic African trypanosomes. Parasitology Research, 95 (3): 186–192.
Njiru: Division of Veterinary and Biomedical Sciences, Western Australian Biomedical Research Institute (WABRI), Murdoch University, South Street, 6150 Murdoch, Western Australia, Australia.
There are 11 different pathogenic trypanosomes in trypanosomiasis endemic regions of Africa. Their detection and characterisation by molecular methods relies on species-specific primers; consequently several PCR tests have to be made on each sample. Here, new primers have been used and the PCR products assessed for specificity and sensitivity against a range of economically and medically important Trypanosoma species. The primers offer promise as a routine diagnostic tool through the use of a single PCR; however, further evaluation is recommended.
13155 Zinsstag, J. & Schelling, E., 2003. Vector-borne diseases in humans and animals: activities of the Swiss Tropical Institute and risks for Switzerland. SAT, Schweizer Archiv für Tierheilkunde, 145 (12): 559–569.
Zinsstag: Swiss Tropical Institute, PO Box, 4002 Basel, Switzerland.
48 2005 Tsetse and Trypanosomiasis Information 6a,b
The recent outbreak of anaplasmosis in a Swiss cattle herd triggered a discussion of the risk of vector-borne diseases in animals and humans in relation to climate changes and other factors. This overview presents the Swiss Tropical Institute’s activities on vector-borne diseases (malaria, trypanosomosis, and leishmaniosis in humans and tick- borne diseases in livestock), describes the possible risks for humans and animals in Switzerland, and discusses options for action in the domains of public health, livestock production and companion animals. Switzerland is increasingly confronted with vector- borne diseases in humans and animals, but this is mainly due to an increase in imported cases. The emergence of a disease in one sector (human or veterinary medicine) may predict future trends in the other. A stronger inter-sectoral collaboration between public health and veterinary institutions at the federal and cantonal level is needed.
(b) PATHOLOGY AND IMMUNOLOGY
[See also 28: 13162]
13156 Abenga, J. N., Sanda, S.A., Idowu, T.B. & Lawani F.A.G., 2002. Effect of acute caprine trypanosomiasis on haemoglobin, urea and serum electrolytes. African Journal of Clinical and Experimental Microbiology, 3 (1): 45–47.
Abenga: Pathology, Epidemiology and Statistics Division, Nigerian Institute for Trypanosomiasis Research, PMB 2077, Kaduna, Kaduna State, Nigeria. [[email protected]]
The effect of acute caprine trypanosomiasis on haemoglobin concentration, urea and serum electrolytes was studied in Red Sokoto goats infected with Trypanosoma vivax. The course of infection lasted only two weeks when the infected goats died of fulminating parasitaemia and high fever. Haemoglobin concentration of the infected goats was only slightly decreased. However, the serum urea level was significantly increased + while Cl, K , and HCO3 levels were slightly increased above pre-infection values by week two post infection. Serum Na+ increased only in the first week post infection, but returned to pre-infection values by the second week.
13157 Al-Qarawi, A.A., Omar, H.M., Abdel-Rahman, H.A., El-Mougy, S.A. & El- Belely, M.S., 2004. Trypanosomiasis-induced infertility in dromedary (Camelus dromedarius) bulls: changes in plasma steroids concentration and semen characteristics. Animal Reproduction Science, 84: (1–2): 73–82.
Al-Qarawi: Department of Veterinary Medicine, Faculty of Agriculture and Veterinary Medicine, King Saud University, P.O. Box 1482, Bureidah, Al- Qassim, Saudi Arabia. [[email protected]]
The effect of trypanosomiasis on concentrations of plasma steroids and semen characteristics was studied in 24 dromedary bulls. Based upon the parasitological and
49 2005 Tsetse and Trypanosomiasis Information 6b,c serological diagnosis, 18 bulls were found infected with Trypanosoma evansi (Group 2) and six were found to be free from infection and served as controls (Group 1). The infected animals exhibited signs of anaemia indicated by the decrease of packed cell volume (PCV) and haemoglobin concentration (Hb), pale mucus membranes, weight loss, lethargy, weakness and dullness. However, five animals (27.8 percent) of the infected group revealed elevated rectal temperatures and three animals (16.7 percent) revealed testicular degeneration upon palpation of their scrotal contents. Concentrations of plasma oestradiol-17ȕ (86.5 ± 8.6 pg/ml versus 232.5 ± 74.4 pg/ml) and testosterone (4.8 ± 0.7 ng/ml versus 2.7 ± 1.5 ng/ml) were significantly different between the control and infected bulls. The semen collected by electroejaculation was evaluated by a computerised cell motion analyzer; this revealed normal semen characteristics in the control animals compared to sub-normal ones in the infected bulls. There were highly significant decreases in sperm count (12.2 ± 1.3/ml versus 6.5 ± 4.9 × 106 ml-1), motility percentage (68.2 ± 6.7 percent versus 27.4 ± 15.6 percent), percentage of live spermatozoa (73.2 ± 8.3 percent versus 35.8 ± 8.2 percent) and increases in percentage of morphological abnormalities (3.3 ± 0.6 percent versus 15.9 ± 1.0 percent) in the infected group. An examination of the plasma hormonal profiles and semen characteristics in the infected bulls indicated that altered Sertoli cell function due to formation of immune complexes in four bulls (Group 2A), pituitary dysfunction in six bulls (Group 2B), testicular degeneration in three bulls (Group 2C) and finally trypanotolerance in five bulls (Group 2D) are possible factors responsible for poor semen characteristics and infertility induced by T. evansi infection in dromedary bulls.
13158 Darsaud, A., Bourdon, L., Chevrier, C., Keita, M., Bouteille, B., Queyroy, A., Canini, F., Cespuglio, R., Dumas, M. & Buguet, A., 2003. Clinical follow- up in the rat experimental model of African trypanosomiasis. Experimental Biology and Medicine, 228 (11): 1355–1362.
Darsaud: Centre de recherches du Service de santé des armées, Département des facteurs humains, La Tronche cedex, France.
Animal models of Human African Trypanosomiasis (HAT) have been developed to understand the pathogenic mechanisms leading to the passage into the neurological phase, most of them referring to histological aspects but not clinical or behavioural data. Our study aimed at defining simple clinical and/or behavioural markers of the passage between the haemolymphatic phase and the meningo-encephalitic stage of the disease. Sprague-Dawley rats (n = 24) were infected with Trypanosoma brucei brucei AnTat 1.1E. Food intake and body weight were measured daily from the day of infection until death. Haematocrit was measured twice a week. Behavioral disturbances were evaluated through an Open-field test. A sudden weight loss occurred on the twelfth day after infection, due to a significant drop of food intake starting two days before. The rats developed an anaemic state shown by the haematocrit measurements. The Open-field test showed them to be less active and reactive as soon as the second week after infestation. A complementary histological study observed trypanosomes and inflammatory cells in the choroid plexus at the same period. These results are in favour of central nervous system functional disturbances. The observed weight loss is discussed as being a measure of the
50 2005 Tsetse and Trypanosomiasis Information 6b,c entry in the meningo-encephalitic phase. The rat model reproduces neurological symptoms observed in the human disease and may prove to be useful for further neurohistological and therapeutic studies.
13159 Mbanasor, U.U., Anene, B.M., Chime, A.B., Nnaji, T.O., Eze, J.I. & Ezekwe, A.G., 2003. Haematology of normal and trypanosome-infected Muturu cattle in southeastern Nigeria. Nigerian Journal of Animal Production, 30 (2): 236– 241.
Mbanasor: Faculty of Veterinary Medicine, University of Nigeria, Nsukka, Nigeria.
Blood parameters of 23 Muturu cattle in a herd were studied between April and August 1998 by monthly examination of their blood samples. Fourteen of a total of 110 blood samples analysed (five samples were unsuitable for analysis) were infected with Trypanosoma vivax. Data from the trypanosome-infected blood were included to evaluate the role of trypanosome infection. The blood values of infected animals were, except for the erythrocyte indices of MCH and MCHC, indistinguishable from those of uninfected animals. There were variations due to age and physiological status (open heifer, pregnant and lactating). Mean RBC and WBC count, MCV and monocyte count were lower in calves than older animals. Open heifers had higher mean RBC and monocyte count, and lower WBC count, MCV and MCH than pregnant and suckling cows.
13160 Shi MeiQing, Wei GuoJian, Pan WanLing & Tabel, H., 2004. Trypanosoma congolense infections: antibody-mediated phagocytosis by Kupffer cells. [mice] Journal of Leukocyte Biology, 76 (2): 399–405.
Tabel: Department of Veterinary Microbiology, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon, Saskatchewan, S7N 5B4, Canada. [[email protected]]
(c) TRYPANOTOLERANCE
[See also 28: 13117, 13147]
13161 Agyemang, K., 2005. Trypanotolerant livestock in the context of trypanosomiasis intervention strategies. PAAT Technical and Scientific Series, 7, pp. viii + 66.
Agyemang: International Trypanotolerance Centre, Banjul, Gambia.
An overview is given of the nature of the tsetse-trypanosomiasis problem in Africa, and the various options available for control. The guiding principles are set out for the role of trypanotolerant livestock within an integrated approach to combating the tsetse and trypanosomiasis problem. From this should come a better understanding of where and when trypanotolerant livestock might be used economically and sustainably to
51 2005 Tsetse and Trypanosomiasis Information 6b,c combat the tsetse and trypanosomiasis problem. The importance of the attitudes of livestock owners is taken into account.
13162 Faye, D., Fall, A., Leak, S., Losson, B. & Geerts, S., 2005. Influence of an experimental Trypanosoma congolense infection and plane of nutrition on milk production and some biochemical parameters in West African Dwarf goats. Acta Tropica, 93 (3): 247–257.
Geerts: Animal Health Department, Institute of Tropical Medicine, Nationalestraat 155, B-2000 Antwerpen, Belgium
The interactions of trypanosomosis and plane of nutrition on health and productivity of multiparous and primiparous West African Dwarf (WAD) does were studied in a multi-factorial experiment including diet (supplementation or basal diet) and infection (infected or control). Experimental does were infected with Trypanosoma congolense at the beginning of the second week post-kidding and monitored for 16 weeks after infection. Trypanosome infection significantly reduced packed cell volume (PCV). Regardless of infection, the drop in PCV from the pre-infection period to the end of the experiment was more severe in animals under restricted diet. Trypanosome parasitaemia tended to be higher in the supplemented group than in the basal diet group and multiparous animals had a higher parasitaemia than primiparous animals. Trypanosome infection as well as dietary supplement had a significant effect on lactation length. Milk off-take from trypanosome-infected does was significantly lower than that from the uninfected control group and there was a positive effect of plane of nutrition. The drop in milk off-take due to trypanosome infection was more severe in the supplemented group than in the group receiving a basal diet due to the number of does from the supplemented group that were withdrawn from the experiment. The effect of trypanosome infection on doe’s live-weight was only noticeable during the first eight weeks of lactation and there was no significant effect on offspring growth rate unless the mother died. Plasma total protein (TP), albumin and cholesterol concentrations were significantly reduced by the infection but were significantly increased by supplementation. Supplemented does had a higher level of cholesterol and a tendency for a higher parasitaemia. Does of high parity also had a higher cholesterol level than primiparous does and, based on the number of animals that were withdrawn from the experiment, they showed a lower resistance to the infection.
13163 Freeman, A.R., Meghen, C.M., MacHugh, D.E., Loftus, R.T., Achukwi, M.D., Bado, A., Sauveroche, B. & Bradley, D.G., 2004. Admixture and diversity in West African cattle populations. Molecular Ecology, 13 (11): 3477–3487.
Freeman: Department of Animal Science and Production and Conway Institute of Biomolecular and Biomedical Research, Faculty of Agriculture, University College Dublin, Belfield, Dublin 4, Ireland.
Genetic analysis of microsatellite variation in 16 West African cattle populations was carried out. West Africa represented a unique juxtaposition of different climatic and
52 2005 Tsetse and Trypanosomiasis Information 6c,d ecological zones in a relatively small geographical area. While more humid coastal regions were inhabited by the tsetse fly, a vector which spread trypanosomiasis among cattle, the disease was not transmitted in the drier areas outside this zone. This was the most thorough study of genetic diversity in cattle within this area, which contained genetically important trypanotolerant cattle breeds. Genetic relationships among the many breeds were examined and levels of diversity were assessed. Admixture levels were determined using a variety of methods. Ancestry informative or population-associated alleles (PAAs) were selected using populations from India, the Near East and Europe. Multivariate analysis, the admix programme and model-based Bayesian admixture analysis approaches were also employed. These analyses showed the direct impact of ecological factors and the profound effect of admixture on the cattle of this region. Results also highlighted the importance of efforts to prevent further dilution of African taurine breeds by zebu cattle.
13164 Hill, E.W., O’Gorman, G.M., Agaba, M., Gibson, J.P., Hanotte, O., Kemp, S.J., Naessens, J., Coussens, P.M. & MacHugh, D.E., 2005. Understanding bovine trypanosomiasis and trypanotolerance: the promise of functional genomics. Veterinary Immunology and Immunopathology, 105 (3–4): 247– 258.
MacHugh: Animal Genomics Laboratory, Department of Animal Science, Conway Institute for Biomolecular and Biomedical Research, Faculty of Agri-Food and the Environment, University College Dublin, Belfield, Dublin 4, Irish Republic.
African bovine trypanosomiasis, caused by the protozoan parasite Trypanosoma congolense, is endemic throughout sub-Saharan Africa and is a major constraint on livestock production. A promising approach to disease control is to understand and exploit naturally evolved trypanotolerance. We describe the first attempt to investigate the transcriptional response of susceptible Boran (Bos indicus) cattle to trypanosome infection via a functional genomics approach using a bovine total leukocyte cDNA microarray platform. Four male Boran cattle were experimentally infected with T. congolense and peripheral blood mononuclear cells (PBMC) were collected before infection and 13, 17, 23 and 30 days post-infection. A reference experimental design was employed using a universal bovine reference RNA pool. Data were normalised to the median of a set of invariant genes (GAPDH) and BRB-Array tools were used to search for statistically significant differentially expressed genes between each time-point. Using a set of 20 microarray hybridizations, we have made a significant contribution to understanding the temporal transcriptional response of bovine PBMC in vivo to a controlled trypanosome infection. The greatest changes were evident 13 days p.i. after parasites were first detected in the blood. Significant differences were observed in clusters of protein kinase C subunits and MHC class I/II related molecules.
(d) TREATMENT
[See also 28: 13135]
53 2005 Tsetse and Trypanosomiasis Information 7b
7. EXPERIMENTAL TRYPANOSOMIASIS
(a) DIAGNOSTICS
13165 Kuboki, N., Sakurai, T., Di Cello, F., Grab, D.J., Suzuki, H., Sugimoto, C. & Igarashi, I., 2003. Loop-mediated isothermal amplification for detection of African trypanosomes. Journal of Clinical Microbiology, 41 (12): 5517–5524.
Sakurai: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080- 8555, Japan. [[email protected]]
While PCR is a method of choice for the detection of African trypanosomes in both humans and animals, the expense of this method negates its use as a diagnostic method for the detection of endemic trypanosomiasis in African countries. The loop-mediated isothermal amplification (LAMP) reaction is a method that amplifies DNA with high specificity, efficiency, and rapidity under isothermal conditions with only simple incubators. An added advantage of LAMP over PCR-based methods is that DNA amplification can be monitored spectrophotometrically and/or with the naked eye without the use of dyes. Here we report our conditions for a highly sensitive, specific, and easy diagnostic assay based on LAMP technology for the detection of parasites in the Trypanosoma brucei group (including T. brucei brucei, T. brucei gambiense, T. brucei rhodesiense, and T. evansi) and T. congolense. We show that the sensitivity of the LAMP-based method for detection of trypanosomes in vitro is up to 100 times higher than that of PCR-based methods. In vivo studies in mice infected with human-infective T. brucei gambiense further highlight the potential clinical importance of LAMP as a diagnostic tool for the identification of African trypanosomiasis.
13166 Njiru, Z.K., Constantine, C.C., Ndung’u, J.M., Robertson, I., Okaye, S., Thompson, R.C.A. & Reid, S.A., 2004. Detection of Trypanosoma evansi in camels using PCR and CATT/T. evansi tests in Kenya. Veterinary Parasitology, 124 (3–4): 187–199.
Njiru: Western Australian Biomedical Research Institute, Division of Health Sciences, School of Veterinary and Biomedical Sciences, Murdoch University, South Street, Murdoch, WA 6150, Australia.
54 2005 Tsetse and Trypanosomiasis Information 7b
(b) PATHOLOGY AND IMMUNOLOGY
[See also 28: 13187]
13167 Abenga, J.N. & Anosa, V.O., 2005. Serum total proteins and creatine levels in experimental Gambian trypanosomosis of vervet monkeys. African Journal of Biotechnology, 4 (2): 187–190.
Abenga: Pathology, Epidemiology and Statistics Division, Nigerian Institute for Trypanosomiasis Research, PMB 2077, Kaduna, Kaduna State, Nigeria. [[email protected]]
Although human African trypanosomosis presently constitutes a major socio- economic problem in several parts of sub-Saharan Africa, conflicting reports on experimental infections appear to be one of the factors limiting the chemotherapeutic control of the disease. An attempt was therefore made to evaluate the effect of two strains of Trypanosoma brucei gambiense on total proteins and other serum biochemical parameters using vervet monkeys as a model. The outcome of both strains in vervet monkeys was traumatic as the monkeys died from infection 12–15 weeks p.i. while the serum proteins increased due to an increase in the serum globulins with resultant fall in the albumin/globulin ratio. Similarly creatinine and fibrinogen levels increased after infection. The study confirms the existence of atypical virulent infections with a resultant early death from T. b. gambiense.
13168 Darsaud, A., Bourdon, L., Mercier, S., Chapotot, F., Bouteille, B., Cespuglio, R. & Buguet, A., 2004. Twenty-four-hour disruption of the sleep-wake cycle and sleep-onset REM-like episodes in a rat model of African trypanosomiasis. Sleep, 27 (1): 42-46.
Darsaud: Centre de recherches du Service de santé des armées, Département des facteurs humains, B.P. 87, F-38702 La Tronche cedex, France.
Patients with human African trypanosomiasis (sleeping sickness) due to the inoculation of Trypanosoma brucei gambiense or T. b. rhodesiense show a major disruption of the 24-hour sleep-wake distribution, accompanied by the occurrence of sleep-onset rapid-eye-movement (REM) sleep episodes, proportional to the severity of the illness. Although animal models of human African trypanosomiasis have been developed to understand the pathogenic mechanisms leading to immune alterations, the development of an animal model featuring the alterations of endogenous biologic rhythms remains a necessity. Following the description of interventions and experiments on the rat infected with T. b. brucei, it is concluded that the Trypanosoma brucei brucei- infected rat is a good model of the syndrome seen in human African trypanosomiasis, ie the 24-hour disruption of the sleep-wake cycle and the occurrence of sleep-onset REM- like sleep episodes.
55 2005 Tsetse and Trypanosomiasis Information 7b
13169 Espinoza, E., Sandoval, E., Mavare, M., González, N. & Rangel, L., 2000. Comparación de la serie eritrocítica y leucocítica en ovejas y cabras infectadas con Trypanosoma vivax. [Comparison of erythrocyte and leucocyte changes in sheep and goats infected with Trypanosoma vivax.] Veterinaria Tropical, 25 (1): 29–39.
Espinoza: Universidad Simón Rodríguez, Rectorado, Apdo. Postal 3690, Caracas, Venezuela.
13170 Faye, D., Sulon, J., Kane, Y., Beckers, J.F., Leak, S., Kaboret, Y., de Sousa, N.M., Losson, B. & Geerts, S., 2004. Effects of an experimental Trypanosoma congolense infection on the reproductive performance of West African Dwarf goats. Theriogenology, 62 (8): 1438–1451.
Geerts: Animal Health Department, Institute of Tropical Medicine, Nationalestraat 155, B-2000, Antwerp, Belgium.
Thirty-six West African Dwarf (WAD) goats were used to assess the effects of an experimental Trypanosoma congolense infection on their reproductive system. Oestrous cycles were synchronised and when confirmed pregnant (n = 31), the does were randomly allocated into control and trypanosome-infected groups. After infection, the animals were carefully observed until parturition. Trypanosome infection caused an increase of rectal temperature, a significant drop in PCV and abortions in 27.8 percent of the infected does. Kids born from infected does had a lower birth weight than kids born from control goats. Eight out of 13 kids (61.5 percent) that were born alive from infected does died during their first week of life. Plasma pregnancy-associated glycoprotein (PAG) and progesterone concentrations were lower in the infected animals than in the controls. In general, PAG concentration in does which aborted dropped before abortion. Our results revealed that artificial T. congolense infection affected reproductive performance of WAD goats with abortions, premature births and perinatal losses being observed. Neither transplacental transmission of T. congolense nor histopathological lesions of the placenta could be demonstrated.
13171 Grab, D.J., Nikolskaia, O., Kim, Y.V., Lonsdale-Eccles, J.D., Ito, S., Hara, T., Fukuma, T., Nyarko, E., Kim, K.J., Stins, M.F., Delannoy, M.J., Rodgers, J. & Kim, K.S., 2004. African trypanosome interactions with an in vitro model of the human blood-brain barrier. Journal of Parasitology, 90 (5): 970– 979.
Grab: Department of Pediatrics, The Johns Hopkins School of Medicine, Baltimore, MA 21287, USA. [[email protected]]
The neurological manifestations of sleeping sickness in man are attributed to the penetration of the blood-brain barrier (BBB) and invasion of the central nervous system by Trypanosoma brucei gambiense and Trypanosoma brucei rhodesiense. However, how African trypanosomes cross the BBB remains an unresolved issue. We have examined the
56 2005 Tsetse and Trypanosomiasis Information 7b traversal of African trypanosomes across the human BBB using an in vitro BBB model system constructed of human brain microvascular endothelial cells (BMECs) grown on Costar Transwell inserts. Human-infective T. b. gambiense strain IL 1852 was found to cross human BMECs far more readily than the animal-infective T. b. brucei strains 427 and TREU 927. Tsetse fly-infective procyclic trypomastigotes did not cross the human BMECs either alone or when coincubated with the bloodstream form T. b. gambiense. After overnight incubation, the integrity of the human BMEC monolayer measured by transendothelial electrical resistance was maintained on the inserts relative to the controls when the endothelial cells were incubated with T. b. brucei. However, decreases in electrical resistance were observed when the BMEC-coated inserts were incubated with T. b. gambiense. Light and electron microscopy studies revealed that T. b. gambiense initially bind at or near intercellular junctions before crossing the BBB paracellularly. This is the first demonstration of paracellular traversal of African trypanosomes across the BBB. Further studies are required to determine the mechanism of BBB traversal by these parasites at the cellular and molecular level.
13172 Mgbenka, B.O. & Ufele, A.N., 2004. Effects of dietary supplementation of vitamin E and selenium on blood parameters of trypanosome-infected rats (Rattus rattus). Bio-research, 2 (1): 8–17.
Mgbenka: Department of Zoology, University of Nigeria, Nsukka, Nigeria.
This study was carried out to evaluate the combined effects of dietary supplementation of vitamin E and selenium on total white blood cells, mononucleated cells, polymorphonucleated cells and packed cell volume of Trypanosoma congolense- infected rats (Rattus rattus). Ninety adult R. rattus were used. The rats were randomly assigned to 6 treatment groups (A–F). Each treatment was replicated three times. The rats were fed chicks’ mash-based diets containing varied quantities of vitamin E and selenium. In treatment A, the control, neither vitamin E nor selenium was included in the diet, and in other treatments (B–F) the chicks’ mash contained different levels of vitamin E and selenium: diet B, 0.1 mg selenium and 60 mg of vitamin E; diet C, 0.3 mg selenium and 80 mg vitamin E; diet D, 0.5 mg selenium and 100 mg vitamin E; diet E, 0.3 mg selenium and 0.0 mg vitamin E; and diet F, 0.0 mg selenium and 80 mg vitamin E. Blood samples of the rats from each treatment were taken on a weekly basis. The experiment lasted five weeks and at the end of which it was found that 0.3 mg selenium and 80 mg vitamin E (diet C) significantly enhanced the resistance of the R. rattus against trypanosomiasis; the rats lived beyond the usual 10–20 days after being infected with T. congolense.
13173 Naessens, J., Kitani, H., Nakamura, Y., Yagi, Y., Sekikawa, K. & Iraqi, F., 2005. TNF-Į mediates the development of anaemia in a murine Trypanosoma brucei rhodesiense infection, but not the anaemia associated with a murine Trypanosoma congolense infection. Clinical and Experimental Immunology, 139 (3): 405–410.
57 2005 Tsetse and Trypanosomiasis Information 7b
Iraqi: International Livestock Research Institute, PO Box 30709, 00100 Nairobi, Kenya.
Development of anaemia in inflammatory diseases is cytokine-mediated. Specifically, the levels of tumour necrosis factor-Į (TNF-Į), produced by activated macrophages, are correlated with severity of disease and anaemia in infections and chronic disease. In African trypanosomiasis, anaemia develops very early in infection around the time when parasites become detectable in the blood. Since the anaemia persists after the first waves of parasitaemia when low numbers of trypanosomes are circulating in the blood, it is generally assumed that anaemia is not directly induced by a parasite factor, but might be cytokine-mediated, as in other cases of anaemia accompanying inflammation. To clarify the role of TNF-Į in the development of anaemia, blood parameters of wild type (TNF-Į+/+), TNF-Į-null (TNF-Į-/-) and TNF-Į- hemizygous (TNF-Į-/+) trypanotolerant mice were compared during infections with the cattle parasite Trypanosoma congolense. No differences in PCV, erythrocyte numbers or haemoglobin were observed between TNF-Į-deficient and wild type mice, suggesting that the decrease in erythrocytes was not mediated by TNF-Į. Erythropoetin (EPO) levels increased during infection and no significant differences in EPO levels were observed between the three mouse strains. In contrast, during an infection with the human pathogen Trypanosoma brucei rhodesiense, the number of red blood cells in TNF-Į-deficient mice remained significantly higher than in the wild type mice. These data suggest that more than one mechanism promotes the development of anaemia associated with trypanosomiasis.
(c) CHEMOTHERAPEUTICS
13174 Campbell, M., Prankerd, R.J., Davie, A.S. & Charman, W.N., 2004. Degradation of berenil (diminazene aceturate) in acidic aqueous solution. Journal of Pharmacy and Pharmacology, 56 (10): 1327–1332.
Campbell: Centre for Drug Candidate Optimisation, Victorian College of Pharmacy, Monash University (Parkville Campus), 381 Royal Parade, Parkville, Victoria 3052, Australia.
The trypanocide berenil was assessed for chemical stability over the pH range 1–8 at 37°C and 0.2 M ionic strength. It was found to be sufficiently unstable under acid conditions that its therapeutic efficacy is most likely severely compromised when administered orally. At pH 3, the half-life was 35 min, decreasing to 1.5 min at pH 1.75. Reaction rate constants were corrected for the effects of buffer catalysis and were found to range from 2.00 min-1 at pH 1 to 6.1 × 10-6 min-1 at pH 8. The pH-rate profile displayed a region (pH 1–4) where specific acid catalysis was dominant, followed by a transitional region (pH 5–7), and finally a region (pH>7) where uncatalysed degradation was most important. It is recommended that berenil be enteric coated for formulations to be used in treating Third World parasitic diseases.
58 2005 Tsetse and Trypanosomiasis Information 7c
13175 Dembitsky, V.M. & Levitsky, D.O., 2004. Arsenolipids. Progress in Lipid Research, 43 (5): 403–448.
Dembitsky: Department of Organic Chemistry, P.O. Box 39231, Hebrew University, Jerusalem 91391, Israel.
Natural arsenolipids are analogues of neutral lipids, like monoglycerides, glycolipids, phospho- and also phosphonolipids. They have been found in microorganisms, fungi, plants, lichens, in marine molluscs, sponges, other invertebrates, and in fish tissues. This review presents structures of natural arsenolipids (and derivatives), their distribution, biogenesis in algae and invertebrates, synthesis, and also biological activity. Arsenolipids are thought to be end products of arsenate detoxification processes, involving reduction and oxidative methylation and adenosylation. The proposed biogenesis of arsenolipids is based on the natural occurrence of arsenic metabolites, and all the intermediates in the proposed pathway have been identified as natural products of algal origin. Different arseno species are shown to be inhibitors of glycerol kinase, bovine carbonic anhydrase, and also are an effective therapy for acute promyelocytic leukaemia, and there has been promising activity noted in other haematologic and solid tumours. Arsonoliposomes demonstrated high anti-trypanosomal activity against Trypanosoma brucei and inhibit growth of some types of cancer cells (HL-60, C6 and GH3).
13176 Fujii, N., Mallari, J.P., Hansell, E.J., Mackey, Z., Doyle, P., Zhou, Y.M., Gut, J., Rosenthal, P.J., McKerrow, J.H. & Guy, R.K., 2005. Discovery of potent thiosemicarbazone inhibitors of rhodesain and cruzain. Bioorganic & Medicinal Chemistry Letters, 15 (1): 121–123.
Guy: Department of Pharmaceutical Chemistry, University of California, San Francisco, CA 94143, USA.
The synthesis and evaluation of a series of thiosemicarbazones as potential inhibitors of cysteine proteases relevant to parasitic diseases are reported. Derivatives of thiosemicarbazone 1 were discovered to be potent inhibitors of cruzain and rhodesain, crucial proteases in the life cycles of Trypanosoma cruzi and T. brucei rhodesiense, the organisms causing Chagas’ disease and sleeping sickness. However, the entire series had only modest potency against falcipain-2, an essential protease for Plasmodium falciparum, the organism causing malaria. Among the active inhibitors, several potently inhibited proliferation of cultures of T. brucei. However, only modest activity was observed in inhibition of proliferation of T. cruzi or P. falciparum.
13177 Huysmans, G., Ranquin, A., Wyns, L., Steyaert, J. & Gelder, P. van, 2005. Encapsulation of therapeutic nucleoside hydrolase in functionalised nanocapsules. Journal of Controlled Release, 102 (1): 171–179.
van Gelder: Department of Molecular and Cellular Interactions, Flanders
59 2005 Tsetse and Trypanosomiasis Information 7c
Interuniversity Institute for Biotechnology (VIB), Free University Brussels, Pleinlaan 2, 1050 Brussels, Belgium.
Liposomes are introduced as encapsulating carriers for prodrug activating enzymes. The technique used is described in detail. The authors state that with these vesicles they have introduced a new strategy for prodrug therapy.
13178 Joubert, K.E., Kettner, F., Lobetti, R.G. & Miller, D.M., 2003. The effects of diminazene aceturate on systemic blood pressure in clinically healthy adult dogs. Journal of the South African Veterinary Association, 74 (3): 69–71.
Joubert: Department of Companion Animal Clinical Studies, Faculty of Veterinary Science, University of Pretoria, Private Bag X04, Onderstepoort, 0110, South Africa.
Diminazene aceturate is a commonly used antibabesial agent. It has been postulated that diminazene may induce a decrease in blood pressure and exacerbate the hypotension presented in dogs with babesiosis. This study was undertaken to assess the effect of diminazene aceturate on the blood pressure of healthy dogs. Six healthy German shepherd dogs between 18 and 24 months of age with a mean weight of 30.4±2.75 kg were used. Blood pressure was directly measured at the following time intervals: -5, 0, 5, 10, 15, 20, 25, 30, 35, 40, 50, 60, 90 and 120 minutes after treatment with diminazene aceturate (4.2 mg/kg) intramuscularly. No statistical difference (P>0.05) was found in blood pressure between any of the time intervals. An increase in heart rate was seen 5 minutes after the administration of diminazene aceturate but no change in blood pressure was evident. This study concluded that diminazene aceturate in its current formulation with antipyrine does not alter blood pressure in healthy adult dogs.
13179 Khabnadideh, S., Pez, D., Musso, A., Brun, R., Pérez, L.M.R., Gonzalez- Pacanowska, D. & Gilbert, I.H., 2005. Design, synthesis and evaluation of 2,4-diaminoquinazolines as inhibitors of trypanosomal and leishmanial dihydrofolate reductase. Bioorganic and Medicinal Chemistry, 13 (7): 2637– 2649.
Gilbert: Welsh School of Pharmacy, Cardiff University, Redwood Building, King Edward VII Avenue, Cardiff, CF10 3XF, UK.
This paper describes the design, synthesis and evaluation of a series of 2,4- diaminoquinazolines as inhibitors of leishmanial and trypanosomal dihydrofolate reductase. Compounds were designed by generating a virtual library of compounds and docking them into the enzyme active site. Following their synthesis, they were found to be potent and selective inhibitors of leishmanial dihydrofolate reductase. The compounds were also found to have potent activity against Trypanosoma brucei rhodesiense, a causative organism of African trypanosomiasis and also against T. cruzi, the causative organism of Chagas’disease. There was significantly lower activity against Leishmania donovani, one of the causative organisms of leishmaniasis.
60 2005 Tsetse and Trypanosomiasis Information 7c
13180 Machocho, A.K., Bastida, J., Codina, C., Viladomat, F., Brun, R. & Chhabra, S.C., 2004. Augustamine type alkaloids from Crinum kirkii. Phytochemistry, 65 (23): 3143–3149.
Chhabra: Department of Chemistry, Kenyatta University, PO Box 43844, 00100 GPO, Nairobi, Kenya.
Sixteen more Amaryllidaceae alkaloids have been isolated from bulbs of Crinum kirkii of which noraugustamine and 4a,N-dedihydronoraugustamine are hitherto unknown. Their structures and those of earlier known alkaloids have been established by physical and spectroscopic analysis. Application of 2D NMR techniques was used for complete characterization of the alkaloids as well as of 3-O-acetylsanguinine. 1,2- diacetyllycorine and 3-O-acetylsanguinine showed activity against Trypanosoma brucei rhodesiense, the parasite associated with sleeping sickness. 3-O-acetylsanguinine also showed some activity against Trypanosoma cruzi.
13181 Magona, J.W., Mayende, J.S.P., Okiria, R. & Okuna, N.M., 2004. Protective efficacy of isometamidium chloride and diminazene aceturate against natural Trypanosoma brucei, Trypanosoma congolense and Trypanosoma vivax infections in cattle under a suppressed tsetse population in Uganda. Onderstepoort Journal of Veterinary Research, 71 (3): 231–237.
Magona: Livestock Health Research Institute, P.O. Box 96, Tororo, Uganda.
The protective efficacy of isometamidium chloride (ISMM) and diminazene aceturate (DIM) against Trypanosoma brucei, Trypanosoma congolense and Trypanosoma vivax infections in cattle under a suppressed tsetse population was assessed in southeast Uganda. A total of 66 and 57 trypanosome-infected cattle were treated with ISMM and DIM respectively, together with 177 trypanosome-free animals not treated, and were followed for 12 months, checking every 4 weeks. There was no statistical difference in the mean time to infection with any trypanosome species in animals treated with ISMM or DIM. However, the mean time to trypanosome infection was significantly longer for treated animals than controls. The mean time to infection with each of the three trypanosome species differed significantly, with the average time to T. vivax infection the lowest, followed by T. congolense and then T. brucei. The protective efficacy of DIM was as good as that of ISMM, implying that curative treatments against trypanosomosis are sufficient for combination with tsetse control. These results indicate that isometamidium chloride and diminazene aceturate have retained their effectiveness against trypanosome infections in southeast Uganda, despite their use over many years.
13182 Mbwambo, Z.H., Apers, S., Moshi, M.J., Kapingu, M.C., Van Miert, S., Claeys, M., Brun, R., Cos, P., Pieters, L. & Vlietinck, A., 2004. Anthranoid compounds with antiprotozoal activity from Vismia orientalis. [T. b. rhodesiense] Planta Medica, 70 (8): 706–710.
61 2005 Tsetse and Trypanosomiasis Information 7c
Mbwambo: Institute of Traditional Medicine, Muhimbili University College of Health Sciences, Dar es Salaam, Tanzania.
A phytochemical investigation of the 80 percent ethanolic extract of stem bark of Vismia orientalis (Guttiferae or Clusiaceae), a plant used in traditional medicine in Tanzania, resulted in the isolation and spectroscopic characterisation of several compounds, of which vismione D exhibited a broad range of antiprotozoal activities including some against Trypanosoma brucei rhodesiense. However, the compound was also slightly cytotoxic against human L6 cells.
13183 Molfetta, F.A., Bruni, A.T., Honorio, K.M. & da Silva, A.B.F., 2005. A structure-activity relationship study of quinine compounds with trypanocidal activity. European Journal of Medicinal Chemistry, 40 (4): 329–338.
da Silva: Departamento de Química e Física Molecular, Instituto de Química de São Carlos, Universidade de São Paulo, CP 780, 13560-970 São Carlos, SP, Brazil.
13184 Moyersoen, J., Choe, J.-W., Fan, E.K., Hol, W.G.J. & Michels, P.A.M., 2004. Biogenesis of peroxisomes and glycosomes: trypanosomatid glycosome assembly is a promising new drug target. FEMS Microbiology Reviews, 28 (5): 603–643.
Michels: Research Unit for Tropical Diseases, Christian de Duve Institute of Cellular Pathology and Laboratory of Biochemistry, Université Catholique de Louvain, ICP-TROP 74.39, Avenue Hippocrate 74, B-1200 Brussels, Belgium. [[email protected]]
In trypanosomatids (Trypanosoma and Leishmania), protozoa responsible for serious diseases of mankind in tropical and subtropical countries, core carbohydrate metabolism including glycolysis is compartmentalised in peculiar peroxisomes called glycosomes. Proper biogenesis of these organelles and the correct sequestering of glycolytic enzymes are essential to these parasites. Biogenesis of glycosomes in trypanosomatids and that of peroxisomes in other eukaryotes, including the human host, occur via homologous processes involving proteins called peroxins, which exert their function through multiple, transient interactions with each other. Decreased expression of peroxins leads to death of trypanosomes. Peroxins show only a low level of sequence conservation. Therefore, it seems feasible to design compounds that will prevent interactions of proteins involved in biogenesis of trypanosomatid glycosomes without interfering with peroxisome formation in the human host cells. Such compounds would be suitable as lead drugs against diseases caused by trypanosomatids.
13185 Nishimura, K., Shima, K., Asakura, M., Ohnishi, Y. & Yamasaki, S., 2005. Effects of heparin administration on Trypanosoma brucei gambiense infection in rats. Journal of Parasitology, 91 (1): 219–222.
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Nishimura: Division of Veterinary Science, Graduate School of Agriculture and Biological Sciences, Osaka Prefecture University, 1-1, Gakuencho, Sakai, Osaka 599-8531, Japan. [[email protected]]
We examined whether heparin administration influences in vivo trypanosome proliferation in infected rats. Administration of heparin every 8 hours via cardiac catheter inhibited growth of Trypanosoma brucei gambiense and prolonged survival of treated rats. Heparin administration increased lipoprotein lipase activity, high-density lipoprotein (HDL) concentration in the blood, and haptoglobin messenger RNA content of the liver. The presence of heparin in culture media did not directly affect proliferation of trypanosomes in vitro. However, the addition of plasma from infected rats treated with heparin to culture media decreased the number of trypanosomes. This effect was decreased by incubating the trypanosomes with benzyl alcohol, a known inhibitor of receptor-mediated endocytosis of lipoprotein. These data suggested that heparin administration reduced the number of trypanosomes in infected rats. Trypanosome lytic factor, a HDL and haptoglobin-related protein, protects humans and some animals from infection by Trypanosoma brucei brucei. In rats, increases in HDL and haptoglobin may affect the proliferation of T. b. gambiense.
13186 Nyasse, B., Ngantchou, I., Tchana, E.M., Sonké, B., Denier, C. & Fontaine, C., 2004. Inhibition of both Trypanosoma brucei bloodstream form and related glycolytic enzymes by a new kolavic acid derivative isolated from Entada abyssinica. Pharmazie, 59 (11): 873–875.
Nyasse: Laboratory of Bio-organic and Medicinal Chemistry, Department of Organic Chemistry, Faculty of Science, University of Yaounde I, Box 812 Yaounde, Cameroon.
13187 Othman, O.E. & Ahmed, S., 2004. Cytogenetic effect of the trypanocidal drug berenil in blood cultures of river buffalo. Journal of Biological Sciences, 4 (2): 180–184.
Othman: Department of Cell Biology, National Research Center, Dokki, Giza, Cairo, Egypt.
The cytogenetic effect of berenil was evaluated at three concentrations (0.25, 0.5, and 1 mg ml-1) on river buffalo lymphocyte cultures in vitro. The formation of chromosomal aberrations, the induction of sister chromatid exchanges, and the measurement of micronuclei formation were the cytogenetic parameters used in the study. The results showed that the number of cells with different types of structural chromosomal aberrations, including breaks, gaps, deletions, fragments and centric fusions, were increased significantly with the three doses of berenil as compared with the control. It is concluded that the three cytogenetic parameters used to evaluate the effect of
63 2005 Tsetse and Trypanosomiasis Information 7c diminazine aceturate (berenil) indicate that this drug has a mutagenic effect on river buffalo lymphocyte cultures.
13188 Salem, M.M. & Werbovetz, K.A., 2005. Antiprotozoal compounds from Psorothamnus polydenius. Journal of Natural Products, 68 (1): 108–111.
Werbovetz: Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, The Ohio State University, Columbus, OH 43210, USA.
Fractionation of the methanolic extract of Psorothamnus polydenius led to the identification of a number of compounds, which were tested in vitro for their antiprotozoal activity against Leishmania donovani and Trypanosoma brucei. Chalcones 1 and 2 and dalrubone exhibited leishmanicidal and trypanocidal properties.
13189 Seebacher, W., Schlapper, C., Brun, R., Kaiser, M., Saf, R. & Weis, R., 2005. Antiprotozoal activities of new bicyclo[2.2.2]octan-2-imines and esters of bicyclo[2.2.2]octan-2-ols. European Journal of Pharmaceutical Sciences, 24 (4): 281–289.
Seebacher: Institute of Pharmaceutical Sciences, Pharmaceutical Chemistry, Karl-Franzens University, Universitätsplatz 1, A-8010 Graz, Austria.
13190 Silva Filho, A.A. da, Albuquerque, S., e Silva, M.L.A., Eberlin, M.N., Tomazela, D.M. & Bastos, J.K., 2004. Tetrahydrofuran lignans from Nectandra megapotamica with trypanocidal activity. Journal of Natural Products, 67 (1): 42–45.
Eberlin: Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Avenida do Café S/N, Ribeirão Preto, 14040- 903, SP, Brazil.
13191 Steverding, D. & Scory, S., 2004. Trypanosoma brucei: unexpected azide sensitivity of bloodstream forms. Journal of Parasitology, 90 (5): 1188–1190.
Steverding: School of Biological Sciences, University of Bristol, Woodland Road, Bristol BS8 1UG, UK. [[email protected]]
Bloodstream forms of Trypanosoma brucei lack cytochromes and are, therefore, insensitive to cyanide. Azide is a toxic anion that bears chemical and biological properties in common with cyanide and may act in a similar way by inhibition of cytochrome c oxidase. It was, therefore, surprising to find that bloodstream forms of T. brucei were sensitive to sodium azide; growth was reduced by 50 percent with 0.1 mM sodium azide. So far, the only enzyme known in bloodstream forms of T. brucei to be sensitive to azide is the iron-containing superoxide dismutase. However, because the activity of the superoxide dismutase was not affected in parasites incubated for 16 h with 0.5 mM sodium azide (a concentration at which no cell proliferates), the toxic action of
64 2005 Tsetse and Trypanosomiasis Information 7c azide cannot be due to inhibition of this enzyme. These results indicate that the general toxicity of azide is different from that of cyanide.
13192 Tanyildizi, S. & Türk, G., 2004. The effects of diminazene aceturate and ceftriaxone on ram sperm. Theriogenology, 61 (2–3): 529–535.
Tanyildizi: Department of Pharmacology and Toxicology, Faculty of Veterinary Medicine, Firat University, 23119 Elazi÷, Turkey.
Effects of diminazene aceturate and ceftriaxone disodium were evaluated on sperm quality of rams. Daily intramuscular injections of diminazene (6 mg/kg) or ceftriaxone (28.5 mg/kg) were given to each of seven Akkaraman rams assigned per drug for two days. Semen samples were collected from the rams at post-treatment 1, 4, 24, 48, 72, 144, 288 and 336 h and examined for sperm characteristics and hyaluronidase activity. Results showed that use of ceftriaxone and diminazene caused significant (P<0.01) decreases in sperm concentration, volume and motility compared to control group within 288 h post- treatment. In addition, hyaluronidase activity increased significantly (P<0.01) in semen of rams treated with ceftriaxone while remained unchanged in those received diminazene. In conclusion, diminazene aceturate and ceftriaxone disodium did not have any deleterious effect on hyaluronidase enzyme. However, both drugs caused impairment of sperm in rams during the 288 h.
13193 Tasdemir, D., Brun, R., Perozzo, R. & Donmez, A.A., 2005. Evaluation of antiprotozoal and plasmodial enoyl-ACP reductase inhibition potential of Turkish medicinal plants. Phytotherapy Research, 19 (2): 162–166.
Tasdemir: Institute of Organic Chemistry, University of Zurich, Winter- thurerstrasse 190, CH-8057 Zürich, Switzerland. [[email protected]]
A total of 58 extracts of different polarity were prepared from various organs of 16 species of Turkish plants and screened for their antitrypanosomal, antileishmanial and antiplasmodial activities. No significant activity was observed against Trypanosoma cruzi, whereas many extracts showed appreciable trypanocidal potential against T. brucei rhodesiense, with the CHCl3-soluble portion of Phlomis kurdica being the most active.
13194 Tasdemir, D., Güner, N.D., Perozzo, R., Brun, R., Dönmez, A.A., Calis, I. & Rüedi, P., 2005. Anti-protozoal and plasmodial FabI enzyme inhibiting metabolites of Scrophularia lepidota roots. Phytochemistry, 66 (3): 355–362.
Tasdemir: Institute of Organic Chemistry, University of Zurich, Winter- thurerstrasse 190, CH-8057 Zürich, Switzerland.
13195 Tsujimura, Y., Watarai, S., Uemura, A., Ohnishi, Y. & Kodama, H., 2005. Effect of anti-ganglioside antibody in experimental Trypanosoma brucei infection in mice. Research in Veterinary Science, 78 (3): 245–247.
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Wataraia: Laboratory of Veterinary Immunology, Division of Veterinary Science, Graduate School of Agriculture and Biological Sciences, Osaka Prefecture University, 1-1 Gakuen-cho, Sakai, Osaka 599-8531, Japan.
The effect of antibody against ganglioside antigen on Trypanosoma brucei parasites was examined in vitro and in vivo using anti-ganglioside GM1 (AGM-1) monoclonal antibody. The antibody showed complement-dependent cytotoxicity against T. brucei with mouse complement. Furthermore, mice given AGM-1 were challenged intraperitoneally with T. brucei. Although all non-treated control mice died within six days after infection, all of AGM-1-injected mice had survived by six days post-infection. These data suggest that antibody against ganglioside antigen on T. brucei has potential in protection against T. brucei infection.
13196 Uemura, A., Watarai, S., Ohnishi, Y. & Kodama, H., 2005. Protective effect of antiganglioside antibodies against experimental Trypanosoma brucei infection in mice. Journal of Parasitology, 91 (1): 73–78.
Watarai: Laboratory of Veterinary Immunology, Division of Veterinary Science, Graduate School of Agriculture and Biological Sciences, Osaka Prefecture University, Sakai, Osaka 599-8531, Japan. [[email protected] u.ac.jp]
Liposome-associated ganglioside antigens (ganglioside GM1 or bovine brain gangliosides) were prepared to evaluate the potential protective efficacy against Trypanosoma brucei. Mice were immunized with liposome-associated ganglioside GM1 or bovine brain gangliosides intraperitoneally (i.p.). After immunization, significantly higher antigen-specific IgG and IgM antibodies were detected in sera than in the non- immunized control group. When sera from immunized mice were analyzed for isotype distribution, antigen-specific IgG1, IgG2a, and IgG3 antibody responses were also noted. After immunization, mice were challenged i.p. with 1 × 102 cells of T. brucei. Sixty percent of liposome-associated ganglioside GM1-immunized mice survived the infection, and all the mice immunized with bovine brain gangliosides-containing liposomes survived. However, all control mice died within seven days after infection. These data demonstrate that liposomes containing ganglioside antigens have the potential usefulness for the induction of a protective immune response against T. brucei infection and suggest the possibility of developing vaccines that may ultimately be used for the prevention of trypanosomiasis.
13197 Verlinde, C.L.M.J., Bressi, J.C., Choe, J., Suresh, S., Buckner, F.S., Van Voorhis, W.C., Michels, P.A.M., Gelb, M.H. & Hol, W.G.J., 2002. Protein structure-based design of anti-protozoal drugs. [T. brucei] Journal of the Brazilian Chemical Society, 13 (6): 843–848.
Verlinde: Biomolecular Structure Center, University of Washington, Box 357742, Seattle, WA98195-7742, USA. [[email protected]]
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The repertoire of drugs to fight protozoal diseases such as malaria, Chagas’ disease, leishmaniasis and African trypanosomiasis is woefully inadequate. Now, genome sequencing and structural genomics projects are quickly elucidating new drug targets, providing incredible opportunities for medicinal chemists. Here, we illustrate the power of structure-based drug design in this process by our efforts to selectively block trypanosomal glycolysis.
13198 Wesongah, J.O., Murilla, G.A., Kibugu, J.K. & Jones, T.W., 2004. Evaluation of isometamidium levels in the serum of sheep and goats after prophylactic treatment against trypanosomosis. Onderstepoort Journal of Veterinary Research, 71 (3): 175–179.
Wesongah: Kenya Trypanosomiasis Research Institute, P.O. Box 362, Kikuyu, Kenya.
Isometamidium chloride has been used for the control of trypanosomosis in animals for over 36 years, but recently there have been reports of prophylaxis failure under natural conditions. In this study, use of the drug for prophylactic purpose against trypanosomosis in small ruminants was investigated. Forty-two sheep and 44 goats were divided into four treatment groups. Groups 1 and 2 were treated with isometamidium chloride (Samorin) at 3-month intervals while groups 3 and 4 were used as controls. All the animals were exposed to natural tsetse challenge and monitored for serum isometamidium levels and anti-trypanosome antibodies. Seven days after drug administration, isometamidium levels were significantly higher in goats 13.7 ± 0.07 ng/ml than in sheep 6.2 ± 0.06 ng/ml. However, the elimination half-life in the sheep was 14.2 ± 0.92 days and was significantly higher than that of the goats 12 ± 0.5 days. This study established that isometamidium metabolism differs between sheep and goats and this difference may have important implications in high tsetse challenge areas.
13199 Witola, W.H., Inoue, N., Ohashi, K. & Onuma, M., 2004. RNA-interference silencing of the adenosine transporter-1 gene in Trypanosoma evansi confers resistance to diminazene aceturate. Experimental Parasitology, 107 (1–2): 47– 57.
Onuma: Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan. [[email protected]]
Drug resistance of trypanosomes is now a problem, but its underlying mechanisms are not fully understood. Cellular uptake of the major trypanocidal drugs is thought to occur through an adenosine transporter. The adenosine transporter-1 gene, TbAT1, encoding a P2-like nucleoside transporter has previously been cloned from Trypanosoma brucei brucei, and when expressed in yeast, it showed very similar substrate specificity to the P2-nucleoside transporter, but could not transport diamidines (pentamidine and diminazene). We have cloned and sequenced a similar gene (TevAT1) from Trypanosoma evansi and found it to have 99.7 percent identity to the TbAT1 gene. To elucidate the role of the TevAT1 gene on diamidine trypanocidal effect, we genetically engineered T. evansi
67 2005 Tsetse and Trypanosomiasis Information 7c for conditional knock-out of the TevAT1 gene by RNA interference (RNAi). Induction of the RNAi resulted in 10-fold depletion of TevAT1 mRNA, with concomitantly significant resistance to diminazene aceturate (berenil). The induced parasites propagated normally and attained peak cell density at an in vitro concentration of berenil, 5.5-fold higher than the IC100 of the wild-type. TevAT1 knock-out had no effect on the trypanocidal activity of suramin and antrycide, but conferred some resistance to samorin. Our findings validate the significance of the TevAT1 adenosine transporter-1 gene in mediating the trypanocidal effect of diamidines in T. evansi. Further, we show for the first time that RNAi gene silencing in T. evansi can be induced using plasmids designed for T. brucei. We also demonstrate the usefulness of real-time PCR in rapidly quantifying mRNA levels in trypanosomes.
8. TRYPANOSOME RESEARCH
(a) CULTIVATION OF TRYPANOSOMES
(b) TAXONOMY, CHARACTERIZATION OF ISOLATES
[See also 28: 13283]
(c) LIFE CYCLE, MORPHOLOGY, BIOCHEMICAL AND MOLECULAR STUDIES
[See also 28: 13124, 13125]
13200 Ackers, J.P., Dhir, V. & Field, M.C., 2005. A bioinformatic analysis of the RAB genes of Trypanosoma brucei. Molecular and Biochemical Parasitology, 141 (1): 89–97.
Field: Wellcome Trust Laboratories for Molecular Parasitology, Department of Biological Sciences, Imperial College, Exhibition Road, London SW7 2AY, UK. [[email protected]]
13201 Alibu, V.P., Storm, L., Haile, S., Clayton, C. & Horn, D., 2004. A doubly inducible system for RNA interference and rapid RNAi plasmid construction in Trypanosoma brucei. Molecular and Biochemical Parasitology, 139 (1): 75–82.
Clayton: Zentrum für Molekulare Biologie der Universität Heidelberg, ZMBH, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany.
13202 Allen, J.W.A., Ginger, M.L. & Ferguson, S.J., 2004. Maturation of the unusual single-cysteine (XXXCH) mitochondrial c-type cytochromes found in trypanosomatids must occur through a novel biogenesis pathway. Biochemical Journal, 383 (3): 537–542.
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Allen: Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK. [[email protected]]
13203 Allen, J.W.A., Ginger, M.L. & Ferguson, S.J., 2005. Complexity and diversity in c-type cytochrome biogenesis systems. Biochemical Society Transactions, 33 (1): 145–146.
Allen: Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK. [[email protected]]
13204 Alsford, S., Glover, L. & Horn, D., 2004. Multiplex analysis of RNA interference defects in Trypanosoma brucei. Molecular and Biochemical Parasitology, 139 (1): 129–132.
Horn: London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT, UK.
13205 Ansede, J.H., Anbazhagan, M., Brun, R., Easterbrook, J.D., Hall, J.E. & Boykin, D.W., 2004. O-alkoxyamidine prodrugs of furamidine: in vitro transport and microsomal metabolism as indicators of in vivo efficacy in a mouse model of Trypanosoma brucei rhodesiense infection. Journal of Medicinal Chemistry, 47 (17): 4335–4338.
Ansede: Division of Drug Delivery and Disposition, School of Pharmacy, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7360, USA.
13206 Baticados, W.N., Witola, W.H., Inoue, N., Kim, J.Y., Kuboki, N., Xuan, X.N., Yokoyama, N. & Sugimoto, C., 2005. Expression of a gene encoding Trypanosoma congolense putative abc1 family protein is developmentally regulated. Journal of Veterinary Medical Science, 67 (2): 157–164.
Inoue: National Research Centre for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido 0808555, Japan.
13207 Becker, M., Aitcheson, N., Byles, E., Wickstead, B., Louis, E. & Rudenko, G., 2004. Isolation of the repertoire of VSG expression site containing telomeres of Trypanosoma brucei 427 using transformation-associated recombination in yeast. Genome Research, 14 (11): 2319–2329.
Rudenko: Peter Medawar Building for Pathogen Research, University of Oxford, Oxford OX1 5SY, UK. [[email protected]]
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13208 Berriman, M., 2004. The genomes of eukaryotic pathogens. South African Journal of Science, 100 (9–10): 452–458.
Berriman: The Wellcome Trust Sanger Institute, Genome Campus, Hinxton, CB10 1SA, UK.
13209 Best, A., Handoko, L., Schlüter, E. & Göringer, H.U., 2005. In vitro synthesised small interfering RNAs elicit RNA interference in African trypanosomes. Journal of Biological Chemistry, 280 (21): 20573–20579.
Göringer: Dept. of Microbiology and Genetics, Darmstadt University of Technology, Schnittspahnstrasse 64287 Darmstadt, Germany. [goringer@ hrzpub.tu-darmstadt.de]
13210 Besteiro, S., Barrett, M.P., Rivière, L. & Bringaud, F., 2005. Energy generation in insect stages of Trypanosoma brucei: metabolism in flux. Trends in Parasitology, 21 (4): 185–191.
Bringaud: Laboratoire de Génomique Fonctionnelle des Trypanosomatides, UMR-5162 CNRS, Université Victor Segalen Bordeaux 2, Bordeaux, France.
The generation of energy in African trypanosomes is a subject of undoubted importance. In bloodstream-form organisms, substrate-level phosphorylation of glucose is sufficient to provide the energy needs of the parasite. The situation in procyclic-form trypanosomes is more complex. For many years, it was accepted that glucose metabolism followed a conventional scheme involving glycolysis, the tricarboxylic acid cycle and ATP-producing oxidative phosphorylation linked to the electron-transport chain. However, progress in sequencing the Trypanosoma brucei genome and the development of gene-knockout and RNA interference technology has provided novel insights. Coupling these new technologies with classical approaches, including NMR and mass spectrometry to analyse glycolytic intermediates and end products, has yielded several surprises. In this article, we summarise how these recent data have helped to change the view of metabolism in procyclic-form T. brucei.
13211 Brems, S., Guilbride, D.L., Gundlesdodjir-Planck, D., Busold, C., Luu, V.-D., Schanne, M., Hoheisel, J. & Clayton, C., 2004. The transcriptomes of Trypanosoma brucei Lister 427 and TREU927 bloodstream and procyclic trypomastigotes. Molecular and Biochemical Parasitology, 139 (2): 163–172.
Clayton: DKFZ, Im Neuenheimer Feld 580, D 69120 Heidelberg, Germany.
13212 Camargo, R.E., Uzcanga, G.L. & Bubis, J., 2004. Isolation of two antigens from Trypanosoma evansi that are partially responsible for its cross-reactivity with Trypanosoma vivax. Veterinary Parasitology, 123 (1–2): 67–81.
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Bubis: Laboratorio de Química de Proteínas, Departamento de Biología Celular, División de Ciencias Biológicas, Universidad Simón Bolívar, Apartado 89.000, Valle de Sartenejas, Baruta, Caracas 1081-A, Venezuela.
13213 Charrière, F., Tan, T.H.P. & Schneider, A., 2005. Mitochondrial initiation factor 2 of Trypanosoma brucei binds imported formylated elongator-type tRNAMet. Journal of Biological Chemistry, 280 (16): 15659–15665.
Schneider: From the Department of Biology/Zoology, University of Fribourg, Chemin du Musée 10, CH-1700 Fribourg, Switzerland. [[email protected]]
13214 Chattopadhyay, A., Jones, N.G., Nietlispach, D., Nielsen, P.R., Voorheis, H.P., Mott, H.R. & Carrington, M., 2005. Structure of the C-terminal domain from Trypanosoma brucei variant surface glycoprotein MITat1.2. Journal of Biological Chemistry, 280 (8): 7228–7235.
Carrington: Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge CB2 1GA, UK. [[email protected]]
13215 Chowdhury, S.F., Guerrero, R.H., Brun, R., Ruiz-Perez, L.M., Pacanowska, D.G. and Gilbert, I.H., 2002. Synthesis and testing of 5-benzyl-2,4- diaminopyrimidines as potential inhibitors of leishmanial and trypanosomal dihydrofolate reductase. [T. brucei] Journal of Enzyme Inhibition and Medicinal Chemistry, 17 (5): 293–302.
Gilbert: Welsh School of Pharmacy, Cardiff University, Redwood Building, King Edward VII Avenue, Cardiff, CF 10 3XF, UK.
13216 Chung Wei-Lien, Carrington, M. & Field, M.C., 2004. Cytoplasmic targeting signals in transmembrane invariant surface glycoproteins of trypanosomes. Journal of Biological Chemistry, 279 (52): 54887–54895.
Field: Department of Pathology, Tennis Court Road, University of Cambridge, Cambridge CB2 1QP, UK. [[email protected]]
13217 Dacks, J.B. & Doolittle, W.F., 2004. Molecular and phylogenetic characterization of syntaxin genes from parasitic protozoa. Molecular and Biochemical Parasitology, 136 (2): 123–136.
Dacks: Department of Zoology, The Natural History Museum, Cromwell Road, London SW7 5BD, UK.
13218 Dardonville, C., Rinaldi, E., Barrett, M.P., Brun, R., Gilbert, I.H. & Hanau, S., 2004. Selective inhibition of Trypanosoma brucei 6-phosphogluconate
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dehydrogenase by high-energy intermediate and transition-state analogues. Journal of Medicinal Chemistry, 47 (13): 3427–3437.
Gilbert: Welsh School of Pharmacy, Redwood Building, Cardiff University, King Edward VII Avenue, Cardiff CF10 3XF, U.K.
13219 Deng, J.P., Schnaufer, A., Salavati, R., Stuart, K.D. & Hol, W.G.J., 2004. High resolution crystal structure of a key editosome enzyme from Trypanosoma brucei: RNA editing ligase 1. Journal of Molecular Biology, 343 (3): 601–613.
Hol: Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, USA. [[email protected]]
13220 Dhir, V., Allen, C.L. & Field, M.C., 2005. Perturbation of local endogenous expression by insertion of Pol I expression constructs into the genome of Trypanosoma brucei. Experimental Parasitology, 109 (3): 198–200.
Field: Department of Biological Sciences, Imperial College, London SW7 2AY, UK. [[email protected]]
13221 Dhir, V., Goulding, D. & Field, M.C., 2004. TbRAB1 and TbRAB2 mediate trafficking through the early secretory pathway of Trypanosoma brucei. Molecular and Biochemical Parasitology, 137 (2): 253–265.
Field: Department of Biological Sciences, Imperial College, London, SW7 2AY, UK. [[email protected]]
13222 DiPaolo, C., Kieft, R., Cross, M. & Sabatini, R., 2005. Regulation of trypanosome DNA glycosylation by a SWI2/SNF2-like protein. Molecular Cell, 17 (3): 441–451.
Sabatini: Global Infectious Diseases Program, Marine Biological Laboratory, Woods Hole, MA 02543 USA. [[email protected]]
13223 Downey, N., Hines, J.C., Sinha, K.M. & Ray, D.S., 2005. Mitochondrial DNA ligases of Trypanosoma brucei. Eukaryotic Cell, 4 (4): 765-774.
Ray: Paul D. Boyer Hall, 611 Charles Young Dr., University of California, Los Angeles, CA 90095-1570 USA. [[email protected]]
13224 Durand-Dubief, M. & Bastin, P., 2003. TbAGO1, an Argonaute protein required for RNA interference, is involved in mitosis and chromosome segregation in Trypanosoma brucei. BMC Biology, 1 (2): pp. (12 December 2003). [see http://www.biomedcentral.com/1741-7007/1/2]
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Bastin: INSERM U565 & CNRS UMR8646, MNHN USM0503, Muséum National d'Histoire Naturelle, 43 rue Cuvier, 75231 Paris cedex 05, France.
13225 Field, M.C., Allen, C.L., Dhir, V., Goulding, D., Hall, B.S., Morgan, G.W., Veazey, P. & Engstler, M., 2004. New approaches to the microscopic imaging of Trypanosoma brucei. Microscopy and Microanalysis, 10 (5): 621– 636.
Field: Department of Biological Sciences, Imperial College, London, SW7 2AY, UK. [[email protected]]
Protozoan parasites are fearsome pathogens responsible for a substantial proportion of human mortality, morbidity, and economic hardship. The principal disease agents are members of the orders Apicomplexa (Plasmodium, Toxoplasma, Eimeria) and Kinetoplastida (trypanosomes, Leishmania). The majority of humans are at risk from infection from one or more of these organisms, with profound effects on the economy, social structure and quality of life in endemic areas; Plasmodium itself accounts for over one million deaths per annum, and an estimated 4 × 107 disability-adjusted life years (DALYs), whereas the Kinetoplastida are responsible for over 100,000 deaths per annum and 4 × 106 DALYs. Current control strategies are failing due to drug resistance and inadequate implementation of existing public health strategies. Trypanosoma brucei, the African trypanosome, has emerged as a favoured model system for the study of basic cell biology in Kinetoplastida, because of several recent technical advances (transfection, inducible expression systems, and RNA interference), and these advantages, together with genome sequencing efforts, are widely anticipated to provide new strategies of therapeutic intervention. Here we describe a suite of methods that have been developed for the microscopic analysis of T. brucei at the light and ultrastructural levels, an essential component of analysis of gene function and hence identification of therapeutic targets.
13226 Figarella, K., Rawer, M., Uzcategui, N.L., Kubata, B.K., Lauber, K., Madeo, F., Wesselborg, S. & Duszenko, M., 2005. Prostaglandin D-2 induces programmed cell death in Trypanosoma brucei bloodstream form. Cell Death and Differentiation, 12 (4): 335–346.
Duszenko: Department of Chemistry and Pharmacy, Interfakultäres Institut für Biochemie, University of Tübingen, Hoppe-Seyler-Str. 4, Tübingen, Germany. [[email protected]]
13227 Gadelha, C., Wickstead, B., Souza, W. de, Gull, K. & Cunha-e-Silva, N., 2005. Cryptic paraflagellar rod in endosymbiont-containing kinetoplastid protozoa. Eukaryotic Cell, 4 (3): 516–525.
Gadelha: Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK. [catarina.gadelha@path.
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ox.ac.uk]
13228 Gao, G.H., Simpson, A.M., Kang, X.D., Rogers, K., Nebohacova, M., Li, F. & Simpson, L., 2005. Functional complementation of Trypanosoma brucei RNA in vitro editing with recombinant RNA ligase. Proceedings of the National Academy of Sciences of the United States of America, 102 (13): 4712–4717.
Simpson: Howard Hughes Medical Institute, University of California, 6780 MRL, 675 Charles E. Young Drive South, Los Angeles, CA 90095, USA. [[email protected]]
13229 Glenn, R.J., Pemberton, A.J., Royle, H.J., Spackman, R.W., Smith, E., Rivett, A.J. & Steverding, D., 2004. Trypanocidal effect of alpha´, beta´- epoxyketones indicates that trypanosomes are particularly sensitive to inhibitors of proteasome trypsin-like activity. International Journal of Antimicrobial Agents, 24 (3): 286–289.
Steverding: School of Biological Sciences, University of Bristol, Woodland Road, Bristol BS8 1UG, UK. [[email protected]]
13230 Greenbaum, D.C., Mackey, Z., Hansell, E., Doyle, P., Gut, J., Caffrey, C.R., Lehrman, J., Rosenthal, P.J. & McKerrow, J.H. & Chibale, K., 2004. Synthesis and structure-activity relationships of parasiticidal thiosemi- carbazone cysteine protease inhibitors against Plasmodium falciparum, Trypanosoma brucei, and Trypanosoma cruzi. Journal of Medicinal Chemistry, 47 (12): 3212–3219.
Greenbaum: Sandler Center for Basic Research in Parasitic Diseases, Department of Pathology, University of California, San Francisco, California 94143, USA.
13231 Guerreiro, L.T.A., Souza, S.S., Wagner, G., De Souza, E.A., Mendes, P.N., Campos, L.M., Barros, L., Pires, P.F., Campos, M.L.M., Grisard, E.C. & Dávila, A.M.R., 2005. Exploring the genome of Trypanosoma vivax through GSS and in silico comparative analysis. Omics, 9 (1): 116–128.
Dávila: Departamento de Bioquímica e Biologia Molecular, Instituto Oswaldo Cruz/Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.
13232 Hall, B., Allen, C.L., Goulding, D. & Field, M.C., 2004. Both of the Rab5 subfamily small GTPases of Trypanosoma brucei are essential and required for endocytosis. Molecular and Biochemical Parasitology, 138 (1): 67–77.
Field: Department of Biological Sciences, Imperial College, Exhibition Road, London SW7 2AY, UK. [[email protected]]
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13233 Hall, B.S., Arun Pal, Goulding, D. & Field, M.C., 2004. Rab4 is an essential regulator of lysosomal trafficking in trypanosomes. Journal of Biological Chemistry, 279 (43): 45047–45056.
Field: Department of Biological Sciences, Imperial College, London SW7 2AY, UK. [[email protected]]
13234 Hall, B.S., Smith, E., Langer, W., Jacobs, L.A., Goulding, D. & Field, M.C., 2005. Developmental variation in Rab11-dependent trafficking in Trypanosoma brucei. Eukaryotic Cell, 4 (5): 971–980.
Field: Department of Pathology, Tennis Court Road, University of Cambridge, Cambridge CB2 1QP, UK. [[email protected]]
13235 Hammarton, T.C., Lillico, S.G., Welburn, S.C. & Mottram, J.C., 2005. Trypanosoma brucei MOB1 is required for accurate and efficient cytokinesis but not for exit from mitosis. Molecular Microbiology, 56 (1): 104–116.
Mottram: Wellcome Centre for Molecular Parasitology, The Anderson College, University of Glasgow, 56 Dumbarton Road, Glasgow G11 6NU, UK. [[email protected]]
13236 Inoue, M., Nakamura, Y., Yasuda, K., Yasaka, N., Hara, T., Schnaufer, A., Stuart, K. & Fukuma, T., 2005. The 14-3-3 proteins of Trypanosoma brucei function in motility, cytokinesis, and cell cycle. Journal of Biological Chemistry, 280 (14): 14085–14096.
Inoue: Department of Parasitology, Kurume University School of Medicine, 67 Asahi-machi, Kurume, Fukuoka 830-0011, Japan.
13237 Janzen, C.J., Lander, F., Dreesen, O. & Cross, G.A.M., 2004. Telomere length regulation and transcriptional silencing in KU80-deficient Trypanosoma brucei. Nucleic Acids Research, 32 (22): 6575–6584.
Cross: Laboratory of Molecular Parasitology, The Rockefeller University, Box 185, 1230 York Avenue, New York, NY 10021-6399, USA. [[email protected]]
13238 Jones, D., Mehlert, A. & Ferguson, M.A.J., 2004. The N-glycan glucosidase system in Trypanosoma brucei. Biochemical Society Transactions, 32 (5): 766–768.
Jones: School of Life Sciences, University of Dundee, Dundee DD1 5EH, UK. [[email protected]]
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13239 Kang, X.D., Rogers, K., Gao, G.H., Falick, A.M., Zhou, S. & Simpson, L., 2005. Reconstitution of uridine-deletion precleaved RNA editing with two recombinant enzymes. [Trypanosomatid mitochondria] Proceedings of the National Academy of Sciences of the United States of America, 102 (4): 1017– 1022.
Simpson: Howard Hughes Medical Institute, University of California, Los Angeles, CA 90095, USA. [[email protected]]
13240 Kao, C.Y. & Read, L.K., 2005. Opposing effects of polyadenylation on the stability of edited and unedited mitochondrial RNAs in Trypanosoma brucei. Molecular and Cellular Biology, 25 (5): 1634–1644.
Read: Department of Microbiology and Immunology and Witebsky Center for Microbial Pathogenesis and Immunology, SUNY Buffalo School of Medicine, Buffalo, New York, USA.
13241 Kessler, P.S. & Parsons, M., 2005. Probing the role of compartmentation of glycolysis in procyclic form Trypanosoma brucei: RNA interference studies of PEX14, hexokinase, and phosphofructokinase. Journal of Biological Chemistry, 280 (10): 9030–9036.
Parsons: Seattle Biomedical Research Institute, Seattle, Washington 98109, USA. [[email protected]]
13242 Kita, K., Nihei, C. & Tomitsuka, E., 2003. Parasite mitochondria as drug target: diversity and dynamic changes during the life cycle. Current Medicinal Chemistry, 10 (23): 2535–2548.
Department of Biomedical Chemistry, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
13243 Lanteri, C.A., Trumpower, B.L., Tidwell, R.R. & Meshnick, S.R., 2004. DB75, a novel trypanocidal agent, disrupts mitochondrial function in Saccharomyces cerevisiae. Antimicrobial Agents and Chemotherapy, 48 (10): 3968–3974.
Tidwell: Department of Pathology and Laboratory Medicine, Room 805, Brinkhous-Bullitt Building, University of North Carolina, Chapel Hill, NC 27599-7525, USA. [Tidwell@ med.unc.edu]
13244 Law, J.A., Huang, C.E., O’Hearn, S.F. & Sollner-Webb, B., 2005. In Trypanosoma brucei RNA editing, band II enables recognition specifically at each step of the U insertion cycle. Molecular and Cellular Biology, 25 (7): 2785–2794.
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Sollner-Webb: Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA. [bsw@ jhmi.edu] .
13245 Laxman, S., Rascón, A. & Beavo, J.A., 2005. Trypanosome cyclic nucleotide phosphodiesterase 2B binds cAMP through its GAF-A domain. Journal of Biological Chemistry, 280 (5): 3771–3779.
Beavo: Department of Pharmacology, University of Washington, Seattle, Washington 98195-7280, USA. [[email protected]]
13246 Lecordier, L., Walgraffe, D., Devaux, S., Poelvoorde, P., Pays, E. & Vanhamme, L., 2005. Trypanosoma brucei RNA interference in the mammalian host. [Mice] Molecular and Biochemical Parasitology, 140 (1): 127–131.
Vanhamme: Laboratory of Molecular Parasitology, Institute for Molecular Biology and Medicine (IBMM), Free University of Brussels, 12 rue des Professeurs Jeener et Brachet, 6041 Gosselies, Belgium. [zknjiru@ hotmail.com]
13247 Lewdorowicz, M., Yoffe, Y., Zuberek, J., Jemielity, J., Stepinski, J., Kierzek, R., Stolarski, R., Shapira, M. & Darzynkiewicz, E., 2004. Chemical synthesis and binding activity of the trypanosomatid cap-4 structure. RNA, 10 (9): 1469–1478.
Darzynkiewicz: Department of Biophysics, Institute of Experimental Physics, Warsaw University, 93 Zwirki and Wigury St., 02-089 Warsaw, Poland [[email protected]]
13248 Liang, X.H., Uliel, S., Hury, A., Barth, S., Doniger, T., Unger, R. & Michaeli, S., 2005. A genome-wide analysis of C/D and H/ACA-like small nucleolar RNAs in Trypanosoma brucei reveals a trypanosome-specific pattern of rRNA modification. RNA, 11 (5): 619–645.
Michaeli: Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan 52900, Israel. [[email protected]]
13249 Liniger, M., Urwyler, S., Studer, E., Oberle, M., Renggli, C. K. & Roditi, I., 2004. Role of the N-terminal domains of EP and GPEET procyclins in membrane targeting and the establishment of midgut infections by Trypanosoma brucei. Molecular and Biochemical Parasitology, 137 (2): 247– 251.
Roditi: Institut für Zellbiologie, Universität Bern, Baltzerstrasse 4, CH-3012 Bern, Switzerland.
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13250 Lowell, J.E. & Cross, G.A.M., 2004. A variant histone H3 is enriched at telomeres in Trypanosoma brucei. Journal of Cell Science, 117 (24): 5937– 5947.
Cross: Laboratory of Molecular Parasitology, The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. [george.cross@ rockefeller.edu]
13251 Mackey, Z.B., O’Brien, T.C., Greenbaum, D.C., Blank, R.B. & McKerrow, J.H., 2004. A cathepsin B-like protease is required for host protein degradation in Trypanosoma brucei. Journal of Biological Chemistry, 279 (46): 48426– 48433.
Mackey: Department of Pathology Tropical Disease Research Unit, University of California, 513 Parnassus HSW 501, San Francisco, CA 94143, USA. [[email protected]]
13252 Mao, J., Gao, Y.G., Odeh, S., Robinson, H., Montalvetti, A., Docampo, R. & Oldfield, E., 2004. Crystallization and preliminary X-ray diffraction study of the farnesyl diphosphate synthase from Trypanosoma brucei. Acta Crystallographica Section D, Biological Crystallography, 60 (10): 1863–1866.
Oldfield: Department of Chemistry and Biophysics, University of Illinois at Urbana-Champaign, 600 South Matthews Avenue, Urbana, IL 61801, USA. [[email protected]]
13253 Matthews, K.R., 2005. The developmental cell biology of Trypanosoma brucei. Journal of Cell Science, 118 (2): 283–290.
Matthews: Institute of Immunology and Infection Research, School of Biological Sciences, University of Edinburgh, West Mains Road, Edinburgh, EH9 3JT, UK. [[email protected]]
Trypanosoma brucei provides an excellent system for studies of many aspects of cell biology, including cell structure and morphology, organelle positioning, cell division and protein trafficking. However, the trypanosome has a complex life cycle in which it must adapt either to the mammalian bloodstream or to different compartments within the tsetse fly. These differentiation events require stage-specific changes to basic cell biological processes and reflect responses to environmental stimuli and programmed differentiation events that must occur within a single cell. The organization of cell structure is fundamental to the trypanosome throughout its life cycle. Modulations of the overall cell morphology and positioning of the specialised mitochondrial genome, flagellum and associated basal body provide the classical descriptions of the different life cycle stages of the parasite. The dependency relationships that govern these morphological changes are now beginning to be understood and their molecular basis
78 2005 Tsetse and Trypanosomiasis Information 8c identified. The overall picture emerging is of a highly organized cell in which the rules established for cell division and morphogenesis in organisms such as yeast and mammalian cells do not necessarily apply. Therefore, understanding the developmental cell biology of the African trypanosome is providing insight into both fundamentally conserved and fundamentally different aspects of the organization of the eukaryotic cell.
13254 Morgan, G.W., Denny, P.W., Vaughan, S., Goulding, D., Jeffries, T.R., Smith, D.F., Gull, K. & Field, M.C., 2005. An evolutionarily conserved coiled-coil protein implicated in polycystic kidney disease is involved in basal body duplication and flagellar biogenesis in Trypanosoma brucei. Molecular and Cellular Biology, 25 (9): 3774–3783.
Field: Department of Pathology, Tennis Court Road, University of Cambridge, Cambridge CB2 1QP, UK. [[email protected]]
13255 Morty, R.E., Shih, A.Y., Fülöp, V. & Andrews, N.W., 2005. Identification of the reactive cysteine residues in oligopeptidase B from Trypanosoma brucei. FEBS Letters, 579 (10): 2191–2196.
Morty: Department of Internal Medicine, University of Giessen School of Medicine, Aulweg 123 (Room 6-11), D-35392 Giessen, Germany
13256 Mussmann, R., Engstler, M., Gerrits, H., Kieft, R., Toaldo, C.B., Onderwater, J., Koerten, H., van Luenen, H.G.A.M. & Borst, P., 2004. Factors affecting the level and localization of the transferrin receptor in Trypanosoma brucei. Journal of Biological Chemistry, 279 (39): 40690– 40698.
Borst: The Netherlands Cancer Institute, Division of Molecular Biology, Plesmanlaan 121, 1066 CX Amsterdam, Netherlands. [[email protected]]
13257 Natto, M.J., Wallace, L.J.M., Candlish, D., Al-Salabi, M.I., Coutts, S.E. & de Koning, H.P., 2005. Trypanosoma brucei: expression of multiple purine transporters prevents the development of allopurinol resistance. Experimental Parasitology, 109 (2): 80–86.
de Koning: Division of Infection and Immunity, Institute of Biomedical and Life Sciences, University of Glasgow, Joseph Black Building, Glasgow G12 8QQ, UK.
13258 Nilsson, D. & Andersson, B., 2005. Strand asymmetry patterns in trypanosomatid parasites. Experimental Parasitology, 109 (3): 143–149.
Andersson: Centre for Genomics and Bioinformatics, Karolinska Institutet, Berzeliusv. 35, SE-171 77 Stockholm, Sweden
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13259 Nyame, A.K., Kawar, Z.S. & Cummings, R.D., 2004. Antigenic glycans in parasitic infections: implications for vaccines and diagnostics. Archives of Biochemistry and Biophysics, 426 (2): 182–200.
Cummings: Department of Biochemistry and Molecular Biology, Oklahoma Center for Medical Glycobiology, University of Oklahoma Health Sciences Center, Biomedical Research Center, Room 417, 975 NE 10th Street, Oklahoma City, OK 73104, USA. [[email protected]]
13260 Palfi, Z., Schimanski, B., Günzl, A., Lücke, S. & Bindereif, A., 2005. U1 small nuclear RNP from Trypanosoma brucei: a minimal U1 snRNA with unusual protein components. Nucleic Acids Research, 33 (8): 2493–2503.
Bindereif: Institut für Biochemie, Justus-Liebig-Universität Giessen Heinrich-Buff-Ring 58, D-35392 Giessen, Germany [albrecht.bindereif@ chemie.bio.uni-giessen.de]
13261 Papageorgiou, I.G., Yakob, L., Al Salabi, M.I., Diallinas, G., Soteriadou, K.P. & De Koning, H.P., 2005. Identification of the first pyrimidine nucleobase transporter in Leishmania: similarities with the Trypanosoma brucei U1 transporter and antileishmanial activity of uracil analogues. Parasitology, 130 (3): 275–283.
De Koning: Institute of Biomedical and Life Sciences, Division of Infection and Immunity, University of Glasgow, Glasgow G12 8QQ, UK.
13262 Peacock, L., Bailey, M. & Gibson, W., 2005. Tetracycline induction of gene expression in Trypanosoma brucei within the tsetse fly vector. Molecular and Biochemical Parasitology, 140 (2): 247–249.
Gibson: School of Biological Sciences, University of Bristol, Woodland Road, Bristol BS8 1UG, UK.
13263 Price, H.P., Panethymitaki, C., Goulding, D. & Smith, D.F., 2005. Functional analysis of TbARL1, an N-myristoylated Golgi protein essential for viability in bloodstream trypanosomes. Journal of Cell Science, 118 (4): 831–841.
Smith: Wellcome Trust Laboratories for Molecular Parasitology, Centre for Molecular Microbiology and Infection, Department of Biological Sciences, Imperial College London, London, SW7 2AZ, UK. [[email protected]]
13264 Raes, G., Brys, L., Dahal, B.K., Brandt, J., Grooten, J., Brombacher, F., Vanham, G., Noël, W., Bogaert, P., Boonefaes, T., Kindt, A., Bergh, R. van den, Leenen, P.J.M., Baetselier, P. de & Ghassabeh, G.H., 2005. Macrophage galactose-type C-type lectins as novel markers for alternatively
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activated macrophages elicited by parasitic infections and allergic airway inflammation. [T. b. brucei] Journal of Leukocyte Biology, 77 (3): 321–327.
Raes: Cellular and Molecular Immunology Unit, Vlaams Interuniversitair Instituut voor Biotechnologie, Vrije Universiteit Brussel, Building E, Level 8, Pleinlaan 2, B-1050 Brussels, Belgium.
13265 Rivière, L., van Weelden, S.W.H., Glass, P., Vegh, P., Coustou, V., Biran, M., van Hellemond, J.J., Bringaud, F., Tielens, A.G.M. & Boshart, M., 2004. Acetyl:succinate CoA-transferase in procyclic Trypanosoma brucei: gene identification and role in carbohydrate metabolism. Journal of Biological Chemistry, 279 (44): 45337–45346.
Boshart: Laboratoire de Genomique Fonctionnelle des Trypanosomatides, UMR-5162 CNRS, Université Victor Segalen Bordeaux II, 33076 Bordeaux Cedex, France. [[email protected]]
13266 Roper, J.R., Guther, M.L.S., MacRae, J.I., Prescott, A.R., Hallyburton, I., Acosta-Serrano, A. & Ferguson, M.A.J., 2005. The suppression of galactose metabolism in procylic form Trypanosoma brucei causes cessation of cell growth and alters procyclin glycoprotein structure and copy number. Journal of Biological Chemistry, 280 (20): 19728–19736.
Ferguson: Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QP, UK. [[email protected]]
13267 Rusconi, F., Durand-Dubief, M. & Bastin, P., 2005. Functional complementation of RNA interference mutants in trypanosomes. BMC Biotechnology, 5 – art. no. 6.
Bastin: UMR5153 CNRS, USM0503 MNHN, U565 INSERM 57, rue Cuvier B.P. 26, F-75231, Paris Cedex 05, France.
13268 Ryan, C.M. & Read, L.K., 2005. UTP-dependent turnover of Trypanosoma brucei mitochondrial mRNA requires UTP polymerization and involves the RET1 TUTase. RNA, 11 (5): 763–773.
Read: Department of Microbiology and Witebsky Center for Microbial Pathogenesis and Immunology, School of Medicine and Biomedical Sciences, State University of New York at Buffalo, Buffalo, NY 14214, USA. [[email protected]]
13269 Sahin, A., Lemercier, G., Tetaud, E., Espiau, B., Myler, P., Stuart, K., Bakalara, N. & Merlin, G., 2004. Trypanosomatid flagellum biogenesis: ARL-3A is involved in several species. Experimental Parasitology, 108 (3–4): 126–133.
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Merlin: Laboratoire de Génomique Fonctionnelle des Trypanosomatides, UMR CNRS 5162, Université de Bordeaux 2, 146 Rue Léo Saignat, 33 000 Bordeaux, France.
13270 Salmon, D., Paturiaux-Hanocq, F., Poelvoorde, P., Vanhamme, L. & Pays, E., 2005. Trypanosoma brucei: growth differences in different mammalian sera are not due to the species-specificity of transferrin. Experimental Parasitology, 109 (3): 188–194.
Pays: Laboratory of Molecular Parasitology, IBMM, Free University of Brussels, 12, rue des Professeurs Jeener et Brachet, B-6041 Gosselies, Belgium.
13271 Schimanski, B., Ma, L.Y. & Günzl, A., 2004. Failure to detect binding of Trypanosoma brucei SNAPc to U2 and U6 snRNA gene sequences by in vitro transcription competition and pull-down assays. Molecular and Biochemical Parasitology, 137 (2): 293–296.
Günzl: Center for Microbial Pathogenesis, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030-3710, USA.
13272 Schlecker, T., Schmidt, A., Dirdjaja, N., Voncken, F., Clayton, C. & Krauth- Siegel, R.L., 2005. Substrate specificity, localization, and essential role of the glutathione peroxidase-type tryparedoxin peroxidases in Trypanosoma brucei. Journal of Biological Chemistry, 280 (15): 14385–14394.
Krauth-Siegel: Biochemie-Zentrum der Universität Heidelberg, Im Neuenheimer Feld 504, 69120 Heidelberg, Germany. [krauth- [email protected]]
13273 Semballa, S., Geffard, M., Daulouede, S., Malvy, D., Veyret, B., Lemesre, J.L., Holzmuller, P., Mnaimneh, S. & Vincendeau, P., 2004. Antibodies directed against nitrosylated neoepitopes in sera of patients with human African trypanosomiasis. Tropical Medicine and International Health, 9 (10): 1104–1110.
Semballa: Laboratoire de Parasitologie, Université de Bordeaux II, Bâtiment 1b, 33076 Bordeaux Cedex, France.
13274 Shi, H.F., Chamond, N., Tschudi, C. & Ullu, E., 2004. Selection and characterization of RNA interference-deficient trypanosomes impaired in target mRNA degradation. [T. brucei] Eukaryotic Cell, 3 (6): 1445–1453.
Ullu: Department of Internal Medicine, Yale Medical School, BCMM
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136D, 295 Congress Ave., New Haven, CT 06536-8012, USA. [[email protected]]
13275 Shi, H.F., Ullu, E. & Tschudi, C., 2004. Function of the trypanosome Argonaute 1 protein in RNA interference requires the N-terminal RGG domain and arginine 735 in the Piwi domain. Journal of Biological Chemistry, 279 (48): 49889–49893.
Tschudi: Department of Epidemiology and Public Health, Yale University Medical School, 295 Congress Ave., New Haven, CT 06536-0812, USA. [[email protected]]
13276 Smith, T.K., Crossman, A., Brimacombe, J.S. & Ferguson, M.A.J., 2004. Chemical validation of GPI biosynthesis as a drug target against African sleeping sickness. EMBO Journal, 23 (23): 4701–4708.
Ferguson: Division of Biological Chemistry and Molecular Microbiology, The Wellcome Trust Biocentre, School of Life Sciences, University of Dundee, Dundee DD1 5EH, UK. [[email protected]]
13277 Stuart, K.D., Schnaufer, A., Ernst, N.L. & Panigrahi, A.K., 2005. Complex management: RNA editing in trypanosomes. Trends in Biochemical Sciences, 30 (2): 97–105.
Stuart: Seattle Biomedical Research Institute, 307 Westlake Avenue North, Suite 500, Seattle, WA 98109 USA. [[email protected]]
13278 Tu, X.M. & Wang, C.C., 2005. Pairwise knockdowns of cdc2-Related Kinases (CRKs) in Trypanosoma brucei identified the CRKs for G1/S and G2/M transitions and demonstrated distinctive cytokinetic regulations between two developmental stages of the organism. Eukaryotic Cell, 4 (4): 755–764.
Wang: Department of Pharmaceutical Chemistry, UCSF, San Francisco, CA 94143-2280 USA. [[email protected]]
13279 Uzcategui, N.L., Szallies, A., Pavlovic-Djuranovic, S., Palmada, M., Figarella, K., Boehmer, C., Lang, F., Beitz, E. & Duszenko, M., 2004. Cloning, heterologous expression, and characterization of three aquaglyceroporins from Trypanosoma brucei. Journal of Biological Chemistry, 279 (41): 42669– 42676.
Duszenko: Physiologisch-chemisches Institut, Universität Tübingen, Hoppe-Seyler Str. 4, 72076 Tübingen, Germany. [micheal.duszenko@uni- tuebingen.de]
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13280 Vondrušková, E., van den Burg, J., Zíková, A., Ernst, N.L., Stuart, K., Benne, R. & Lukeš, J., 2005. RNA interference analyses suggest a transcript- specific regulatory role for mitochondrial RNA-binding proteins MRP1 and MRP2 in RNA editing and other RNA processing in Trypanosoma brucei. Journal of Biological Chemistry, 280 (4): 2429–2438.
Lukes: Institute of Parasitology, Czeck Academy of Sciences, University of South Bohemia, 37005 Ceské BudƟjovice, Czech Republic. [jula@ paru.cas.cz]
13281 Walgraffe, D., Devaux, S., Lecordier, L., Dierick, J.-F., Dieu, M., Abbeele, J. van den, Pays, E. & Vanhamme, L., 2004. Characterization of subunits of the RNA polymerase I complex in Trypanosoma brucei. Molecular and Biochemical Parasitology, 139 (2): 249–260.
Pays: Laboratory of Molecular Parasitology, Department of Molecular Biology, IBMM, Free University of Brussels, 12, rue des Professeurs Jeener et Brachet, B-6041 Gosselies, Belgium.
13282 Walker, R., Jr., Saha, L., Hill, G.C. & Chaudhuri, M., 2004. The effect of over-expression of the alternative oxidase in the procyclic forms of Trypanosoma brucei. Molecular and Biochemical Parasitology, 139 (2): 153– 162.
Chaudhuri: Department of Microbiology, Meharry Medical College, Nashville, TN 37208, USA.
13283 Witola, W.H., Sarataphan, N., Inoue, N., Ohashi, K. & Onuma, M., 2005. Genetic variability in ESAG6 genes among Trypanosoma evansi isolates and in comparison to other trypanozoon members. Acta Tropica, 93 (1): 63–73.
Onuma: Laboratory of Infectious Diseases, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.
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