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Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
DNA barcoding for disease DNA barcoding for disease vectors vectors
Daniel Adjei BoakyeDaniel Adjei BoakyeParasitology DepartmentParasitology Department
Noguchi Memorial Institute for Medical Noguchi Memorial Institute for Medical ResearchResearch
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
Lymphatic filariasisLymphatic filariasis
elephantiasiselephantiasis
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
OnchocerciasisOnchocerciasis
Health and Socio-Health and Socio-economic impacteconomic impact
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
LeishmaniasisLeishmaniasis
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
Impact of parasitic diseasesImpact of parasitic diseases
Cause immense sufferingCause immense suffering Life-long disabilitiesLife-long disabilities Impair childhood growth and developmentImpair childhood growth and development Promote poverty, impair education andPromote poverty, impair education and Economic development Economic development
• Arable land desertedArable land deserted Population affectedPopulation affected MortalityMortality
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
Neglected Tropical Diseases and MalariaNeglected Tropical Diseases and MalariaDiseaseDisease VBDVBD
11 Buruli ulcerBuruli ulcer ??
22 ChagasChagas **
33 CholeraCholera ****
44 DengueDengue **
55 DracunculiasisDracunculiasis **
66 Human African Human African TrypanosomiasisTrypanosomiasis
**
77 LeishmaniasisLeishmaniasis **
88 LeprosyLeprosy
99 FilariasisFilariasis **
1010 OnchocerciasisOnchocerciasis **
1111 SchistosomiasisSchistosomiasis **
1212 Soil-Transmitted Soil-Transmitted HelminthiasisHelminthiasis
1313 TrachomaTrachoma ****
1414 MalariaMalaria **
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
DiseasesDiseases VectorsVectors AreasAreas
ChagasChagas TriatomaTriatoma S/AmericaS/America
DengueDengue MosquitoMosquito
(Ae)(Ae)
Tropical WorldTropical World
Guinea wormGuinea worm CyclopCyclop AfricaAfrica
H/TrypanosomiasisH/Trypanosomiasis GlossinaGlossina AfricaAfrica
LeishmaniasisLeishmaniasis PhlebotomusPhlebotomus Africa, America,Africa, America,
AsiaAsia
Lymphatic filariasisLymphatic filariasis MosquitoMosquito
(An,Cx,Ae,Mn)(An,Cx,Ae,Mn)
Tropical WorldTropical World
OnchocerciasisOnchocerciasis SimuliumSimulium Africa, AmericaAfrica, America
SchistosomiasisSchistosomiasis SnailSnail Tropical WorldTropical World
MalariaMalaria MosquitoMosquito
(An)(An)
Tropical WorldTropical World
Vectors of Neglected Tropical Diseases
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
Importance of correct identificationImportance of correct identification
Understanding the diversity of vector Understanding the diversity of vector species and their respective vectorial species and their respective vectorial importance is necessary for any importance is necessary for any targeted control efforttargeted control effort
Studies on Vector speciesStudies on Vector species• Identification and clarification of species Identification and clarification of species
statusstatus• Biology and ecology of vectorsBiology and ecology of vectors• Vectorial importance of different speciesVectorial importance of different species
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
Members of the Members of the S. damnosumS. damnosum complex in West Africa complex in West Africa
Determination of important vector Determination of important vector speciesspecies
Basis for mapping limits of control Basis for mapping limits of control programmeprogramme
Only larvae can be identified with Only larvae can be identified with precisionprecision
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
Anopheles gambiaeAnopheles gambiae species complex species complex
Anopheles gambiae sensu lato
An. gambaie s.s
An. arabiensis
An. melas An. merus
An. quadrannulatus
An. bwambae
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
Taxonomic challenges within Taxonomic challenges within An. gambiaeAn. gambiae s.s s.s
Genetic discontinuitiesGenetic discontinuities• from chromosomal from chromosomal
analysis (Coluzzi analysis (Coluzzi et alet al 1985)1985)
Five chromosomal forms Five chromosomal forms in West Africa: Mopti; in West Africa: Mopti; Bamako; Bissau; Forest; Bamako; Bissau; Forest; SavannaSavanna
• in the ribosomal DNA in the ribosomal DNA region (rDNA) (Favia region (rDNA) (Favia et alet al 1997)1997)
Two molecular forms Two molecular forms described (della Torre et described (della Torre et al 2001; 2005)al 2001; 2005)
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
Problems associated with current Problems associated with current identificationidentification
In Mali and Burkina Faso, the M and S forms In Mali and Burkina Faso, the M and S forms correspond to the Mopti and Savanna correspond to the Mopti and Savanna chromosomal forms chromosomal forms
Molecular forms do not always correspond to the Molecular forms do not always correspond to the chromosomal forms in other countrieschromosomal forms in other countries
The two molecular forms may be a reflection of The two molecular forms may be a reflection of the presence of two different species in what is the presence of two different species in what is considered as one species – considered as one species – An. gambiaeAn. gambiae s.s. s.s.
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
Practical consequences for disease Practical consequences for disease control control
Susceptibility to Susceptibility to insecticidesinsecticides
Differences in disease Differences in disease transmission transmission capabilitiescapabilities
Adaptation to different Adaptation to different breeding sitesbreeding sites
There is therefore the There is therefore the need to understand need to understand whether there are 2 or whether there are 2 or only 1 speciesonly 1 species - 5 - 4 - 3 - 2 - 1 0 1 2 3 4
5
6
7
8
9
10
11
12
13
14
15
1 0
2
2 2
3 0
5
1 5
3 2
1 0
5
1 0
6
2
50
1 6
7
1 2
1 7
9
1 0
6 2
5
3 4
4 85 02 6
5
7 4
7 2
1 5
3 0
6 0
7 2
5 0
4 4
3
03
8
2 2
4
0
6
6
6
2
5 4
5
4 4
9
6
4 0
4 31 0
2 6
3
1 2
8
1 305
4 6
1 66
1 4
1 7
4
52 74
9
0
6
1 3
0
4
1 6
1
1 22 01 8
0
8
3
6
7
2 1
53
3 2
6 0
5
6
1 39
2 8
0
3
4 8
0
9
7
4 2
5
4
3
3 9
5 8
0
4
4
3
5 23 4
1
7
8
3 8
5
4 2
2 8
2 2
9
6
05
0
4 4
0
3
4 0
9
3 0
1
2
65
5
3 2
6 0
2 4
0
1
0
3 2
5 2
24
0
5 2
6
0
0
1 1
4
6
4 0
2
2 1
60
8
1 9
1
6
2
1 2
3
3 4
2
6 21 9
0
6
0
24
98
4 6
0
2
2
2
8
4 6
1 1
1 5
31
1 2
1 4
3 1
2 4
6
1 0
3 5
54
6
3 3
4 6
0
5 2
6 8
1 8
1 45 4
6
8
0
3
6
1 2
3 5
4 8
0
2 8
3 4
1 7
5 8
2
79
1 21 02 7
50
3 2
1 4
1 0
1 1
2
1 0
5 2
0
7
3 5
1 4
0
5
0
0
0
7
1 0
3
6 6
4 8
0
3 8
43
2 0
9
3 6
0
4 8
6 0
0
0
0
9
1
0
2
0
0
3 2
1
3 6
0
0
6 0
00
0
0
5 8
0
0
0
0
0
0
72 6
0
5 6
3
0
0
0
00
1
0
4 0
1
4
1 1
0
00
0
0
001
3
4
0
0
1
0
0
3 1
00
0
000
1
00
0
00
0
0
0
0
0
0
00
0
0
3 8
0
0
0
6
5 4
6
0
00
51 07
7
4
4 6
0
0
3
1
1
2
0
3
1
0
0
0
0
0
0
7
1
1 1
0
0
1 3
0
2
94
0
4
0
00
0
2
0
0
1
0
0
0
0
0
1
0
0
Distribution of LF in Burkina Faso, Benin, Togo and Ghana
E stim a tedp rev a len ceo f ag (% )
0
1
2
3
5
10
30
50
B u rk in a F aso
B en in
T o g oG h an a
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
MethodologyMethodology
Multiplex PCR for species
identification PCR – RFLP for molecular form identification
Morphological identification -
Gillies and Coetzee (1987)
Intergenic spacer of the rDNA region of the X-chromosome Scott et al. (1993), Favia et al. (1997) and Fanello et al (2002)
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
MethodologyMethodology
PCR of CO1 fragment & Sequencing:
(Applied Biosystems 377 automated DNA
sequencer)
Sequence alignment & comparisons: The sequences obtained were aligned using BioEdit (Hall, 1999) BLAST database search for closely related
sequences in the Genebank database . Analysis for species relationships (Kimura 2-parameter (K2P) model; Kimura, 1980: Neighbour-Joining (NJ) tree;
Saitou & Nei, 1987) were conducted in Mega 4 (Tamura et al., 2007).
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
Preliminary resultsPreliminary results
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
Preliminary resultsPreliminary results
Molecular Molecular forms of An. forms of An. gambiae s.s.gambiae s.s.
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
Example of phylogenetic treeExample of phylogenetic tree
TE6
TE8
TE9
TE3
TE1
TE2
39 Ankwanda melas
41 Ankwanda melas
40 Ankwanda gambiae ss M form
17 Ankwanda gambiae ss S form
35 Ankwanda gambiae ss M form
69 (nice peaks) Ada melas
23 Ada gambiae ss M form
KP7 Kpone on sea gambiae ss S form
WA33
WA44
AK130
SA2
SA4
SA3
SA1
SA10
0.02
Sudan Savanna
Coastal Savanna
Semi Deciduous Forest
Savanna
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
Ghana Mosquitoes’ file in the Completed Project section of the Barcode of Life Ghana Mosquitoes’ file in the Completed Project section of the Barcode of Life
website (website (http://http://www.barcodinglife.orgwww.barcodinglife.org).).
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
ConclusionConclusion
DNA barcode can be used to DNA barcode can be used to • Separate out closely related members of Separate out closely related members of
sibling species in vectors of diseasessibling species in vectors of diseases• Draw relationships between species Draw relationships between species
collected from wider geographical areascollected from wider geographical areas• Determine the presence of individuals Determine the presence of individuals
currently unidentified using the currently unidentified using the available techniquesavailable techniques
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
AcknowledgementsAcknowledgements
Funding: Funding: • JRS Biodiversity FoundationJRS Biodiversity Foundation• World Health OrganizationWorld Health Organization• Gate’s FoundationGate’s Foundation
Collaborators at the NMIMRCollaborators at the NMIMR• Prof. Michael WilsonProf. Michael Wilson• Dr Maxwell AppawuDr Maxwell Appawu• Dziedzom de SouzaDziedzom de Souza• Hilaria AmuzuHilaria Amuzu• Ignatius NdongIgnatius Ndong
Western and Central Africa DNA BarcodingWestern and Central Africa DNA Barcoding meeting: Abuja, Nigeria meeting: Abuja, Nigeria
Thank youThank you