1. Eastern Africa Barcode Workshop, Oct 18-19 DNA Barcoding - Parasites and Vectors Dan Masiga Molecular Biology and Biotechnology Department

2. Eastern Africa Barcode Workshop, Oct 18-19 Insects

3. Eastern Africa Barcode Workshop, Oct 18-19 Vector-transmitted pathogens Viruses Rickettsia Bacteria Protozoa helminths

4. Eastern Africa Barcode Workshop, Oct 18-19 TDR Disease Focus 1.African trypanosomiasis 2.Chagas disease 3.Leishmaniasis 4.Dengue 5.Lymphatic filariasis 6.Malaria 7.Onchocerciasis 8.Schistosomiasis 9.Tuberculosis 10.Leprosy UNICEF/UNDP/World Bank/WHO - TDR http://www.who.int/tdr/grants/strategic-emphases/files/matrix.pdf TDR Disease Focus 1.African trypanosomiasis 2.Chagas disease 3.Leishmaniasis TDR Disease Focus 1.African trypanosomiasis 2.Chagas disease 3.Leishmaniasis 4.Dengue 5.Lymphatic filariasis 6.Malaria 7.Onchocerciasis 4.Dengue 5.Lymphatic filariasis 6.Malaria

5. Eastern Africa Barcode Workshop, Oct 18-19 Distribution of human-infective trypanosomes causing sleeping sickness Animal-infective trypanosomes Are found throughout sub-saharan Africa, with the exception of the Southern tip.

6. Eastern Africa Barcode Workshop, Oct 18-19 Well fed tsetse Barcoding tsetse flies and trypanosomes T. brucei in blood smear tsetse fly feeding

7. Eastern Africa Barcode Workshop, Oct 18-19 Tsetse flies and trypanosomes There are about 31 species of tsetse flies, the only members of genus Glossina, are recognized. About 10 tsetse-transmitted trypanosome species and sub-species are known; probably >100 other species. Beyond Africa, T. evansi and T. vivax are transmitted mechanically (without parasite development in the vector)

8. Eastern Africa Barcode Workshop, Oct 18-19

9. Loci that have been used Tsetse:ITS 1 Trypanosomes:Satellite DNA; small-subunit ribosomal RNA (ssu rRNA) gene Barcoding tsetse and trypanosomes

10. Eastern Africa Barcode Workshop, Oct 18-19 Identification of trypanosomes based on satellite DNA Satellite DNA is defined as “a portion of DNA in eukaryotes whose density differs from that of the majority of DNA and that consists of short, repeating sequences of nucleotide pairs (typically 5-500 bp long).

11. Eastern Africa Barcode Workshop, Oct 18-19 Trypanosome phylogeny (and ID) using ssu rDNA (18S rDNA locus)

12. Eastern Africa Barcode Workshop, Oct 18-19 Current phylogeny is based on 18S rDNA phylogeny is based on 18S rDNA (taken from Hamilton et al., 2004)

13. Eastern Africa Barcode Workshop, Oct 18-19 Can COI be used for barcoding trypanosomatids?

14. Eastern Africa Barcode Workshop, Oct 18-19 mitochondrion glycosomes Inside a trypanosome

15. Eastern Africa Barcode Workshop, Oct 18-19 Kinetoplast DNA (mitochondrial DNA) 20-50 identical maxicircles (20-40 kb) encode 18 protein encoding (12 non- functional) genes, 2 rRNAs and a few guide (g) RNAs. 5000-12000 heterogeneous minicircles encode (over 900 different) small guide RNAs. Maxi- and minicircles are interlocked in a giant catenate. Dyskinetoplastic cells lack minicircles, maxicircles or both, e.g. T. evansi. Cannot be transmitted by Tsetse. EM of the periphery of an isolated kDNA network Network of minicircles condensed network Prof. Christine Clayton

16. Eastern Africa Barcode Workshop, Oct 18-19 Some benefits of using COI A single locus of accessible size. Resolve classification of Glossina at sub-generic level (3 or 4 have been proposed: fusca, palpalis, austeni & morsitans). A more accurate indication of relationships and distances, perhaps providing explaining some key phenotypes, e.g. transmission efficiencies.

17. Eastern Africa Barcode Workshop, Oct 18-19 Sandflies, vectors of Leishmaniasis Species of Leishmania L. major L. aethiopica L. infantum L. donovani L. tropica L. mexicana complex L. guyanensis L. naiffi L. braziliensis L. tarentolae E. scheideri Lutzomyia spp. (New World ) Phlebotomus spp. (Old World) Lutzomyia longipalpis lapinha TDR image Lib ID 00061063

18. Eastern Africa Barcode Workshop, Oct 18-19 Insects

19. Eastern Africa Barcode Workshop, Oct 18-19 Barcoding and epidemiology/disease control

20. Eastern Africa Barcode Workshop, Oct 18-19 Global view of endemic regions for leishmaniasis With such a wide distribution, you can start small, but must network to advance.

21. Eastern Africa Barcode Workshop, Oct 18-19 NHGRI Awards $13M for Next Generation Sequencing Technologies [October 4, 2006] NEW YORK (GenomeWeb News) – The National Human Genome Research Institute has awarded eleven grants worth a total of more than $13 million to develop technologies that will lower the cost of DNA sequencing. The agency has a near-term goal of lowering the costs of sequencing a mammalian-sized genome to $100,000, and to eventually cut the cost of whole- genome sequencing to $1,000 or less. Lowering the cost will make genome sequencing possible as part of routine medical care, NHGRI said. Nine grants fund researchers developing technologies that will make the $1,000 genome possible and two grants for researchers working on the $100,000 mammalian genome: