Aedes aegypti control programmes in Brazil. Margareth L. Capurro

Transcription

1 Aedes aegypti control programmes in Brazil Margareth L. Capurro

2 Emerging Infectious Diseases Vol. 12, No. 6, June 2006

3 Integrate Control for Aedes aegypti Population Supression

4 Integrate Control for Aedes aegypti Population Supression

5 Integrate Control for Aedes aegypti Population Supression

6 Integrate Control for Aedes aegypti Population Supression 80%

7 Integrate Control for Aedes aegypti Population Supression 80% 80%

8 Integrate Control for Aedes aegypti Population Supression 80% 80%

9 Step 1. Suppression Natural Population Step 2. Replacement

10 Open Field Release of OX513A Aedes aegypti Transgenic line evaluation

11 Repressive of Insects carrying a Dominant Lethal gene (RIDL) Antidote (Tetra) X teto Minimal promoter tta death Thomas et al Science 287:

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13 Males for releases 1,5 million/week

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15 Project Phase 2 Jacobina - Bahia

16 Pupa Transportation

17 Pupa transportation (LEMI) C25 BOD 16 o C ON 180,000 per container Arriving at LEMI Emergency, Monitoring and Information Lab Preparation for release

18 First Trial Second Trial

19 What do we learn? We need Genetic Sexing Strain (GSS) Producing Sterile male strain (no Larvae) Use of tetracycline only in colonies

20 Sterility Conditional Construct - SCC No Antidote Antidote (Tet +) X No Viable Eggs Viable Eggs

21 SCC Transgenes Two Effector molecules: Endonuclease 3XP3 DsRed tto Endonuclease B2tubulin ttav SV40 IAP Antagonist 3XP3 DsRed tto Michelob_X B2tubulin ttav SV40

22 Transgenic Lines 150 pb 350 pb 150 pb

23 Sterile Conditional Construct (SCC) 60% Homozygous 80% Reduction

24 Improving transgenic lines Aedes aegypti and Aedes albopictus Producing Sterile male strain (no Larvae) Producing GSS (Genetic Sexing Strain)

25 Dsx knockout

26 Step 1. Suppression Natural Population Step 2. Replacement

27 Gene Introduction Virus-regulated mosquito gene Suicidal Model (Double death model) Natural Population

28 Gene Introduction Virus-regulated mosquito gene Suicidal Model (Double death model) Natural Population + Introduction

29 Gene Introduction Virus-regulated mosquito gene Suicidal Model (Double death model) Virus Infection Natural Population + Introduction

30 Gene Introduction Virus-regulated mosquito gene Suicidal Model (Double death model) Virus Infection Natural Population + Introduction

31 Gene Introduction Virus-regulated mosquito gene Suicidal Model (Double death model) Step 2. Replacement NS3 Cleavage site RRRRSAG HS-PRO Constitutive Promoter Mx Lethal protein Sec61γ ER localization Lúmen do RER C-terminal L4 L2 L6 L8 Sec61γ Sec61β

32 Cleavage sites A5 RRRRSAG RRRRSAG HS EGFP Mx Sec pr M E NS1 ER Lumen SAG RRRR C 4 2A 2B NS3 4A 4B 3 NS5 Cytoplasm 5 6 X or APOPTOSIS Double death model - Infection phenotypes in dengue challenged mosquito?

33 C6/36 cells - Transfection and D2 challenge: GFP filter Light GFP filter Light DAPI Merge DAPI Merge PIE EGFP Sec61 Mx EGFP GFP filter Light 3 days after DEN2 GFP filter Light ER marker red dye Merge DAPI Merge Sec61 Mx EGFP Sec61 Mx EGFP

34 Intrathoracic injection - Survivorship Percent survival Wild type CS (n=77) Wild type IS (n=82) p= Days after injection Percent survival DUI-Y CS (n=81) DUI-Y IS (n=66) p= Days after injection Survival rates of wild type (Higgs-strain) and DUI-Y transgenic line challenged with DENV-2 via intrathoracic injection. Five-to-seven day old females were injected with control supernatant (CS) and DENV-2 infected supernatant (IS) (vial load = 7.0 x 10 6 genomic copies). Females were recovered during 24hs after injection and survived females were maintained on 10% sucrose solution and censored during 14 days after injection (14DPI). Solid lines are showing survival curves of females injected with CS and dotted lines are showing survival curves of females injected with IS. Survival curves were statistically analyzed by Log-rank (Mantel-Cox) test.

35 Bringing new technology to the field Technical Community Male Regulator y Nature

36 How the Sterility works? How the Sterile Mosquito Works? Why releasing male mosquitoes you kill mosquitoes? Males Female Wild Sterile X

37 Before releasing mosquitoes Site selection Public Engagement: Evolving Public Authorities (Government and agencies); Local ones (community engagement); Local people - explain what we are going to do in that area.

38 Community Engagement

39 Community Engagement Total people 17,101,269 in Brazil Based on the Brazilian Institute of Public Opinion and Statistics (IBOPE) data

40 City Hall Public Hearing Talks and Lectures

41 Leaflet distribution

42 Mosquito Aedes \Dengue Muriçoca (Culex) Pica durante o dia (bite during the day) Pica durante a noite (bite during the night)

43 Bar Blood for Sale! Only females (girls) bite Nectar Fonte: Google

44 ACTION TARGET POPULATION LEVEL # EVENTS # PEOPLE Presentations/Lectures Local/Regional Leaflets ( ¹ ) Local - 10,000 Jingle (¹) Local - - Meetings National/ International 39 6,020 Interviews (radio) Regional 15 1,500 Interviews (TV) Regional/National 09 17,094,000 (2) Interviews Local/Regional/ (newspaper/magazine) National 13 - Internet (website / social network) Regional/National 24 - ( ³ ) Houses visited/interviewed with residents Local 581 2,341 Meetings with local leaders, health agents Local Presentations at elementary and middle school Local Presentation at community center/city hall/others Local Driving truck with loudspeakers in the releasing area Local Spots, jingles and short messages Local broadcasted in local radio station 52 1,200 (4) TOTAL 17,101,269

45 STRATEGIES Mandatory Recommended Suggested - Visit/interview sample/every house in the target area - Meetings with local leaders, school principals, district managers - Lectures at schools targeting kids/teens - Press coverage at local/regional level of PAT activities: production, releases - Lectures at community centers/churches targeting adults - Radio spots, jingles and messages broadcasted - Press releases by Moscamed journalists - PAT technical personnel interviewed by local/regional/(inter)national radio stations - Press coverage at national level of PAT activities: production, releases - Action within a local event (parade, carnival, street fairs) - Driving truck with loudspeakers in the targeting area jingle and messages - Use of social media: Facebook and twitter - Press coverage at international level of PAT activities: releases

46 Spot and Jingle

47 SPOT To control dengue Moscamed is releasing in this community A large amount of TRANSGENIC MOSQUITOES. We would like to recall that this mosquitoes are not the well known CULEX They are transgenic MALES and they DON T BITE. They are good fellows that will give you protection against dengue. For more information call a health agent or get in touch with MOSCAMED By the phone (74) PAT AEDES TRANSGENIC PROJECT This one makes the difference.

48 Jingle Transgenic Aedes

49 Universidade de São Paulo UPAT LEMI Dr Peter Piermarini Dr Jeff Powell Dr Lincoln Suesdek Dr Chun-Hong Chen Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular