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Tick Research at ILRI 
Richard Bishop 
8 International Congress on Ticks and Tick-Borne Pathogens (TTP8) 
and 12 Biennial Conference, South Africa, 24-29 August 2014
ILRI Tick Unit Workshop 
ICTTD Cape Town 2014 
• How Does ILRI Reintegrate into 
the Global Tick Research Arena? 
• How can we better utilise the ILRI 
Tick unit -Collaborative projects? 
•
ILRI Tick Unit- Tick Species 
• Ixodid ticks (9 species) 
• Rhipicephalus appendiculatus: 9 stocks 
Vector of ECF- differences in vector 
competence for Theileria parva 
• R. zambeziensis, R.evertsi, R.pulchellus 
• Amblyomma variegatum 
• A.gemma 
• Hyalomma anatolicum 
• Boophilus (Rhipicephalus) decoloratus 
• B.(Rhipicephalus) microplus
Tick Unit facilities and expertise 
• Capacity for 16 cattle experiments for 
evaluation of vaccines or acaricides 
• Rabbit room with capacity for 48 rabbits 
• Salivary gland and other tissue 
dissection 
• RNAi equipment 
• Manager- with an agenda to develop s 
bew scientific activities-Naftaly Githaka
Laboratory Capacity 
• Advanced Molecular biology 
infrastructure 
• Next Generation sequencing 
- 454 titanium 
- Illumina Mi Seq 
Library preparation specialist: 
Bioinformatics Cluster 
Systems Administrator
The First Commercial ‘Anti-Parasite’ Vaccine 
TickGard (based on 
recombinant BM86 gut 
glycoprotein) from 
Rhipicephalus (Boophilus) 
microplus
BM86 and homologues 
• BM86 protects against Boophilus (Rhipicephalus ) 
decoloratus but not R. appendiculatus 
• Homologues cloned from R. decoloratus and R. 
appendiculatus- 
• In three host tick R. appendiculatus moulting from 
nymph to adults was affected 
• Protection does not correlate with sequence identity- 
Better efficacy with heterologus challenge 
• Conserved epitopes identified-Pepscan Hylomma- 
Rhipicephalus- Boophilus
ILRI-Evaluation of Unconventional 
tick vaccine candidates in cattle 
• Cement proteins 
• RIM 36-Immunodominant in cattle- no protection 
• Trp64-from IxodesPat Nuttall group protects against 
human encephalitis virus- no protection in cattle 
• Histamine Binding (Lipocalin fold) protein-ambiguous 
results
Genomics and Functional genomics for novel 
vaccine candidates 
• Salivary gland EST databases developed in 
collaboration with TIGR 
• Amblyomma variegatum 
• R.appendiculatus-infected versus uninfected- 
Many differences (7000+) low abundance 
• Suppression subtractive approach to look for 
transcripts up-regulated in infected salivary 
glands
Tick Genomics 
• Genome organisation. 
• Many Transposons 
• Novel SINE element 
• Allelic exclusion of very distinct RA86 
variants in R. appendulatus lab stock 
• Development of microsatellite markers 
for R. appendiculatus laboratory and 
field populations
A Wider Research Agenda 
• Recent Review of ILRI tick unit by Peter 
Willadsen-Ex CSIRO-recommended inclusion of 
novel activities in additional tick species 
• Activities related to direct tick control- Evaluating 
vaccines and acaricides-Group 1 
• Tick-pathogen interaction research-Group2 
• Measuring tick burdens and impact –Group 3 
Special focus on B. (R) microplus in Africa
ECF recombinant vaccine support 
• Sporozoite production 
• Tick challenge model-All current work is based on ITM (live 
sporozoite and drug administration model) 
• Investigate mechanisms of immunity from tick-delivered T. parva 
• Tick-sporozoite antigen combination vaccines 
• Complexity of challenge scenario-taxonomy of ticks using new 
techniques and monitoring pathogen loads
ITM improvement irradiated sporozoite 
vaccine 
• Irradiated sporozoite vaccine 
• Plasmodium-Malaria analogy 
• Not fully researched for T. parva 
• Interest from Sanaria 
• Simplify composition of current vaccine.
Improved tick control 
• Novel acaricide evaluation: 
- Sigma-Genosys acaricide trial 
New Vaccines- 
• FER2 evaluation as a vaccine candidate in cattle for 
b. microplus 
• Participation in wider evaluation of novel candidates 
from reverse vaccinology
Acaricide Resistance Monitoring 
• In Collaboration with Ministries of Livestock 
• Not Well researched in Africa 
• Start in East Africa
Tick-Host-Pathogen interface 
Theileria parva 
Arthropod 
vector 
Pathogen-Host 
Infection of vertebrate host 
Role of tick salivary gland factors? 
Parasite evasion of vertebrate host 
Vertebrate host 
(i) 
(iii) 
(ii) 
Pathogen-vector 
Pathogen has to overcome tick ‘barriers’: 
1. Gut Invasion 
2. Tick immune system 
3 Salivary gland maturation 
Tick- vertebrate host 
responses 
Prolonged attachment by Ixodid tick: 
Tick injects a cocktail of peptides and lipids 
Tissue cement to enhance feeding 
Inflammatory responses 
Immunomodulation 
Haemostatic responses
ILRI Nascent Activities for 
tick/pathogen interaction 
• Two R. appendiculatus genetically related 
stocks that differ in vector competence for T. 
parva 
• Metagenomics- Novel viral pathogen 
discovery-Access to African tick species
Livestock Tick distribution and 
Impact 
• Special focus on Boophilus (Rhipicephalus) in 
an African context- This tick is spreading 
rapidly in South, East and West Africa 
• Test for acaricide resistance not well 
monitored in Africa 
• What is the direct impact of ticks on livestock 
production including milk?
Tick Challenge Model 
• Efficacy of anti-pathogen or anti-vector antigens in the 
field is difficult to test using a needle challenge in the 
laboratory 
• Needle challenge is a MODEL SYSTEM 
• Therefore an experimental tick challenge system run in 
parallel will be very valuable 
• Baseline data on tick challenge has been generated at 
ILRI using a selected low abundance line-designed to 
be more comparable to field challenge
R. appendiculatus salivary gland EST 
Data from T.parva infected and 
uninfected females 
• 18,000+ total sequences RA sequences. 
7359 unique-Generated in collaboration with 
TIGR. 
• No Major clusters qualitatively different 
between T. parva infected and uninfected 
salivary glands
ILRI research-Combination 
sporozoite-tick salivary gland 
• T.parva p67 sporozoite antigen better in the 
laboratory than the field 
• Combine with salivary gland antigens 
• Antibody responses can be induced against 
seven distinct recombinant antigens P67, and 
six salivary gland components
Better lives through livestock 
ilri.org 
The presentation has a Creative Commons licence. You are free to re-use or distribute this work, provided credit is given to ILRI.

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Tick research at ILRI

  • 1. Tick Research at ILRI Richard Bishop 8 International Congress on Ticks and Tick-Borne Pathogens (TTP8) and 12 Biennial Conference, South Africa, 24-29 August 2014
  • 2. ILRI Tick Unit Workshop ICTTD Cape Town 2014 • How Does ILRI Reintegrate into the Global Tick Research Arena? • How can we better utilise the ILRI Tick unit -Collaborative projects? •
  • 3. ILRI Tick Unit- Tick Species • Ixodid ticks (9 species) • Rhipicephalus appendiculatus: 9 stocks Vector of ECF- differences in vector competence for Theileria parva • R. zambeziensis, R.evertsi, R.pulchellus • Amblyomma variegatum • A.gemma • Hyalomma anatolicum • Boophilus (Rhipicephalus) decoloratus • B.(Rhipicephalus) microplus
  • 4. Tick Unit facilities and expertise • Capacity for 16 cattle experiments for evaluation of vaccines or acaricides • Rabbit room with capacity for 48 rabbits • Salivary gland and other tissue dissection • RNAi equipment • Manager- with an agenda to develop s bew scientific activities-Naftaly Githaka
  • 5. Laboratory Capacity • Advanced Molecular biology infrastructure • Next Generation sequencing - 454 titanium - Illumina Mi Seq Library preparation specialist: Bioinformatics Cluster Systems Administrator
  • 6. The First Commercial ‘Anti-Parasite’ Vaccine TickGard (based on recombinant BM86 gut glycoprotein) from Rhipicephalus (Boophilus) microplus
  • 7. BM86 and homologues • BM86 protects against Boophilus (Rhipicephalus ) decoloratus but not R. appendiculatus • Homologues cloned from R. decoloratus and R. appendiculatus- • In three host tick R. appendiculatus moulting from nymph to adults was affected • Protection does not correlate with sequence identity- Better efficacy with heterologus challenge • Conserved epitopes identified-Pepscan Hylomma- Rhipicephalus- Boophilus
  • 8. ILRI-Evaluation of Unconventional tick vaccine candidates in cattle • Cement proteins • RIM 36-Immunodominant in cattle- no protection • Trp64-from IxodesPat Nuttall group protects against human encephalitis virus- no protection in cattle • Histamine Binding (Lipocalin fold) protein-ambiguous results
  • 9. Genomics and Functional genomics for novel vaccine candidates • Salivary gland EST databases developed in collaboration with TIGR • Amblyomma variegatum • R.appendiculatus-infected versus uninfected- Many differences (7000+) low abundance • Suppression subtractive approach to look for transcripts up-regulated in infected salivary glands
  • 10. Tick Genomics • Genome organisation. • Many Transposons • Novel SINE element • Allelic exclusion of very distinct RA86 variants in R. appendulatus lab stock • Development of microsatellite markers for R. appendiculatus laboratory and field populations
  • 11. A Wider Research Agenda • Recent Review of ILRI tick unit by Peter Willadsen-Ex CSIRO-recommended inclusion of novel activities in additional tick species • Activities related to direct tick control- Evaluating vaccines and acaricides-Group 1 • Tick-pathogen interaction research-Group2 • Measuring tick burdens and impact –Group 3 Special focus on B. (R) microplus in Africa
  • 12. ECF recombinant vaccine support • Sporozoite production • Tick challenge model-All current work is based on ITM (live sporozoite and drug administration model) • Investigate mechanisms of immunity from tick-delivered T. parva • Tick-sporozoite antigen combination vaccines • Complexity of challenge scenario-taxonomy of ticks using new techniques and monitoring pathogen loads
  • 13. ITM improvement irradiated sporozoite vaccine • Irradiated sporozoite vaccine • Plasmodium-Malaria analogy • Not fully researched for T. parva • Interest from Sanaria • Simplify composition of current vaccine.
  • 14. Improved tick control • Novel acaricide evaluation: - Sigma-Genosys acaricide trial New Vaccines- • FER2 evaluation as a vaccine candidate in cattle for b. microplus • Participation in wider evaluation of novel candidates from reverse vaccinology
  • 15. Acaricide Resistance Monitoring • In Collaboration with Ministries of Livestock • Not Well researched in Africa • Start in East Africa
  • 16. Tick-Host-Pathogen interface Theileria parva Arthropod vector Pathogen-Host Infection of vertebrate host Role of tick salivary gland factors? Parasite evasion of vertebrate host Vertebrate host (i) (iii) (ii) Pathogen-vector Pathogen has to overcome tick ‘barriers’: 1. Gut Invasion 2. Tick immune system 3 Salivary gland maturation Tick- vertebrate host responses Prolonged attachment by Ixodid tick: Tick injects a cocktail of peptides and lipids Tissue cement to enhance feeding Inflammatory responses Immunomodulation Haemostatic responses
  • 17. ILRI Nascent Activities for tick/pathogen interaction • Two R. appendiculatus genetically related stocks that differ in vector competence for T. parva • Metagenomics- Novel viral pathogen discovery-Access to African tick species
  • 18. Livestock Tick distribution and Impact • Special focus on Boophilus (Rhipicephalus) in an African context- This tick is spreading rapidly in South, East and West Africa • Test for acaricide resistance not well monitored in Africa • What is the direct impact of ticks on livestock production including milk?
  • 19. Tick Challenge Model • Efficacy of anti-pathogen or anti-vector antigens in the field is difficult to test using a needle challenge in the laboratory • Needle challenge is a MODEL SYSTEM • Therefore an experimental tick challenge system run in parallel will be very valuable • Baseline data on tick challenge has been generated at ILRI using a selected low abundance line-designed to be more comparable to field challenge
  • 20. R. appendiculatus salivary gland EST Data from T.parva infected and uninfected females • 18,000+ total sequences RA sequences. 7359 unique-Generated in collaboration with TIGR. • No Major clusters qualitatively different between T. parva infected and uninfected salivary glands
  • 21. ILRI research-Combination sporozoite-tick salivary gland • T.parva p67 sporozoite antigen better in the laboratory than the field • Combine with salivary gland antigens • Antibody responses can be induced against seven distinct recombinant antigens P67, and six salivary gland components
  • 22. Better lives through livestock ilri.org The presentation has a Creative Commons licence. You are free to re-use or distribute this work, provided credit is given to ILRI.