CDC ENTOMOLOGY BRANCH ACTIVITIES

12 February 2007

Department of Health and Human Services (HHS)

US Public Health Service (USPHS) National Institutes of Health (NIH) Centers for Disease Control and Prevention (CDC) National Center for Zoonotic, Vector-borne & Enteric Diseases (NCZVED) – Formally NCID

ENTOMOLOGY BRANCH

Diseases Activities

• • • • • • • •

Malaria Chagas disease Lymphatic filariasis Onchocerciasis Leishmaniasis West Nile virus Trachoma Ectoparasites

• • • • • •

Outbreak investigations Intervention: ITN, IRS, drugs Insecticide resistance Molecular & vector biology Surveillance & training OCONUS: 15 PMI countries, Guatemala, Indonesia and DoD labs

World’s Most Dangerous Animals

Vector-Borne Diseases Impact

Disease (2001) Malaria Lymphatic filariasis Afr. trypanosomiasis Disease Burden DALYs (Thousands) 40,213 5,549 1,810 Deaths (Thousands) 1,080 0 41 Leishmaniasis Onchocerciasis Chagas disease Dengue TOTAL 1,713 951 680 433 51,349

DALYS = disability adjusted life years; Data from the WHO

11 0 21 12 1,165

Entomology Branch Personnel

16 FTEs + ~30 fellows, students, visiting scientists, military entomologists & others 8 FTE Entomologists: Lived overseas >50 years; field and laboratory TDY >15 yrs in more than 80 countries 2 FTE Chemists: Analytical and biological analysis of drugs and insecticides

Insectaries & Malaria Research and Reference Reagent Resource (MR4) Center

• • •

Insectaries - Total ~280 sq m (~2,500 sq ft) 12 separate rooms (two for infected vectors) Biosphere building ~90 sq m (~1,000 sq ft) Produce >3.5 million mosquitoes/year, to include all primary vectors of human malaria

• •

MR4 Vector Reagents Living Anopheles stocks (~50 species/strains) Field specimens/genomic-DNA; LN2 ~80,000

WHO Collaborating Centers

•

Malaria Vector Identification - ELISA

•

Insecticide Resistance in Vectors

•

Evaluation of Existing and New Insecticides

•

Evaluation of Anti-malarial Drugs (pending)

•

Malaria Control (MB)

•

Control/Elimination LF in the Americas (PDB)

•

Human African Trypanosomiasis (PDB)

Malaria: Worldwide

~500 million new infections annually; >1.4 million/day >1 million deaths/yr; a child dies every 20-30 sec A disease of the poor, ~60% in poorest ~20% Increasing problem in cities, now with ~15% of burden 1-4% loss in African GDP; ~US$12B/yr Over 40% worlds’ population is at risk ~10,000 travelers from Europe, Japan and NA (~2,000) contract malaria each year No vaccine; effective drugs expensive/counterfeited Resistant: parasites to drugs & vectors to insecticides

Malaria models for evaluation of drugs, vaccines and vector competency

Dr. Collins – 53 years at CDC; “…a national treasure…”

Identification of Malaria Vectors

• Only ~40 of ~430

Anopheles

species are important vectors of human malarias • Vector collection and ID a challenge

Need Replacement for HLC

Human landing collections

Primary Malaria Vector Complexes

Vector Competency/Parasite Interactions

ELISA: >2M VecTest: >45K

Molecular Lab: pop’n genetics & transgenics Germ line transformation of An. gambiae with GFP-transposable element

Insecticide Resistance Surveillance Program

• LARVAL & ADULT BIOASSAYS • BIOCHEMICAL ASSAYS • MOLECULAR ASSAYS • MOSQUITO POOLS • FIELD STUDIES AND TRAINING

ADULT RESISTANCE BOTTLE BIOASSAY

Bottle treated with insecticide only Susceptible population Test population

MICRO-PLATE BIOCHEMICAL ASSAYS

100 90 80 70 60 50 40 30 20 10 0 0.1

0.3

0.5

0.7

0.9

1.2

1.4

Absorbance 570 nm 1.6

1.8

2 Susceptible Resistant

MOLECULAR ASSAYS: Measure insecticide resistance genes…….

Copy Number Expression Level Point Mutations

EB Chemistry Activity

•

Anti-malarial drug analysis

•

Resistance vs. compliance

•

Pharmacokinetics

•

QA of pharmaceuticals

•

Counterfeit drug detection

•

Insecticide/formulation testing

•

ITN efficacy evaluations

•

New ITN treatment methods

Analytical Testing Capability for all Available Anti-Malarial Drugs Quinoline compounds: Quinine/ Quinidine Chloroquine Amodiaquine Mefloquine Halofantrine Primaquine Antifol combinations: Pyrimethamine/ sulfadoxine Pyrimethamine/ sulfalene Trimethoprim/ sulfamethoxazole Biguanides and Biguanide/ sulfa combinations: Proguanil Proguanil/ sulfone Miscellaneous: Tetracyclines Clindamycin Atovaquone (+ Proguanil) Pyronaridine Azithromycin Artemisinins Benflumetol (+ Artemether)

Malaria Therapy Drug Costs

Chloroquine or SP Mefloquine Artemisinins Malarone US$ 0.10

US$ 3.00

US$ 3.70

US$ 8.00

Assay: <10 minutes; >99% of tablet available for use 2001-4: No artesunate in 38% OTC tablets in Vietnam, Laos, and Myanmar; ~50% OTC Cambodia counterfeit Africa with expensive combination therapy = ?

GC Analysis ITN Bioassay

Laboratory Evaluation of LLTNs

100 80 60 40 20 0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Number of Washes 17 18 R 19 eg en 20 er at io n Pe rmaNe t® 1.0

Dawa® Inse ctor Olyse t® Cyclode xtrin Conv e ntional

DPD Domestic Malaria Activities • Prevention & Treatments Guidelines – Drug options, other prevention options – Malaria case management in the United States • Prevention Communication – “Health Information for International Travel” – Building and updating interactive WEB pages • Assist states: lab support & outbreak investigations

CDC Assists States in Malaria Outbreaks: 63 in 59 years (1957-2006)

1 5 6 17 2 3 4 Routes of Migration

Laboratory Diagnosis of Malaria

Microscopic Based Diagnosis Giemsa Stained Morphology PCR based Diagnosis rRNA Genes

QuickTime™ and a Photo - JP EG decompressor are needed to see this picture.

Molecular Analysis of PBC P. vivax

• • •

MSP-3

a/-b

& CSP VK210 genes identical by sequence or RFLP.

S-type rRNA and ORF 470 genes of New World type Conclusion: All 8 P. vivax infections likely originated from a single source of infection from Latin America PBC_MSP3B(S) PBC_MSP3B(F) PBC_MSP3B(MOC) Ao.1

67B.1

67T.5

56 IEC.2

Belem IEC.8

VA_KM3B VA_JB3B VA_NF3B 69.1

India 781B SalI Chess BellT NK Thai 781T

MSP-3 b Phenogram

Cey57 SL.3

Bangl

CDC OCONUS Malaria Activities

Amazon Malaria Initiative LSTM GF PMI CDC/KEMRI CDC TZ India Indonesia PMI: ANG, BEN, TAZ, UGA, SEN, MOZ, GAH, MALI, LIB, ETH, RWA KEN, ZAM, MAD, MLWI

CDC person in country CDC Regional projects PMI supported programs Single country support programs

Amazon Malaria Initiative (AMI)

CDC assists with surveillance, treatment, drug resistance, vector control & training

Partners: USAID, PAHO, NMRCD, CDC and MOHs Countries: Bolivia, Brazil, Colombia, Ecuador, Peru, Guyana, Suriname, Venezuela

CDC Entomology Support in Indonesia

# # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # #

CDC entomological support =

High Medium Low

None Level of malaria transmission

CDC Malaria Activities: Indonesia

• ~2 million cases & 40,000 deaths annually • Range of transmission intensities & ecologies • Support from partners, e.g., USAID, UNICEF, GF • Opportunities to learn how to drive transmission to ever lower levels — relevance to Africa • Post-tsunami entomological evaluation & now have a Ent Br FTE entomologist in Indonesia

President’s Malaria Initiative (PMI)

A 5-year, $1.2 B initiative to rapidly scale-up malaria control interventions in high burden countries in Africa.

Goal: Reduce malaria-related mortality by 50% in 15 countries

PMI Malaria Control Tools

•

Transmission reduction – insecticide treated nets (ITNs), indoor residual spraying (IRS) & larval control

•

Case management using rapid diagnostic tests (RDT) & artemisinin combination treatment (ACT)

•

Intermittent Preventive Treatment in pregnancy (IPTp)

•

CDC has made important contributions to developing, evaluating and improving these strategies

CDC/KEMRI Field Station Western Kenya Gem Asembo CDC/KEMRI laboratoryy at Kisian Equator

Provincial Hospital Kisumu Lake Victoria

KENYA

KENYA ITN SUMMARY: 120,000 people

ITNs Mosquitoes 38%



47%



35%



28%



Parasitemia Severe anemia Placental malaria Low birth weight 95% fewer An gambiae Healthier pregnancies 90% reduction in transmission 74%



Healthier children Malaria attack rate Median time to 1st infection 4.5



10.7 mo 60%



Incidence of severe anemia Improved infant and child growth 27%



Sick child visits to the clinic Improved infant and child survival 26%



14%



Infant mortality Child (1-4yr) mortality

Field Evaluation of LLTNs – Essential for PMI

1 0.9

0.8

0.7

0.6

0.5

0.4

0.3

0.2

0.1

0 0 Cyclodextrin Olyset Insector PermaNet DAWA Conventional 30 60 90 120 150 180 210 240 270 300 330 360 390 420 450 480 510 540 570 600 630 660 690 720

Days since distribution

*Failure defined as 2 consecutive bioassays with <50% mortality

Indoor Residual Spraying

Entire household protected No change in human behavior required

•

Coordinate IRS with MOH & contractor

•

Evaluate insecticides & formulations

•

Evaluate more efficient application methods, e.g. ULV

Photo courtesy of Dr Mark Benedict

Experimental Hut DDT Studies Ifakara - March 07

• Brazil =

An. darlingi

– Monitor entering, exiting & biting.

– Sprayed vs. unsprayed houses – Spatial Repellent action stopped >95% from entering house – Behavioral actions> 12 months • Tanzania =

An. gambiae

Insecticide Based Control

• •

Other materials eave curtains, calendars/posters

•

Excito-repellents & MOA studies Goal: Efficacy of IRS & distribution ease of ITNs

Chagas Disease

Cases: 16-18 million Deaths: 50,000 /yr At risk: 90 million; 21 countries Control: Three multi-national control programs (Southern Cone, Andean Pact, and CA) Agent: Trypanosoma cruzi Vector: Triatomine bugs Transmission: Vector-borne >80% Transfusion ~16% No effective treatment for established infections New drugs needed

Chagas Disease Insecticide-Based Vector Control Obstacles



Incomplete domestic coverage



Reinfestation by peridomestic/ sylvatic populations (T. dimidiata in CA; T. sordida and T. brasiliensis in SA)



Insufficient post-treatment surveillance



Effects on non-target insects



Cost and long term sustainability

Molecular Approach: Bacterial symbiont Rhodococcus rhodnii and Chagas parasite Trypanosoma cruzi live in the gut of Rhodnius prolixus bugs

Symbionts transmitted to nymphs by coprophagy - the ingestion of feces

Insects not acquiring the symbionts die

Genetically Modified (GM) Bacterial Symbionts

(Transmission) Cure Insect of symbiont Isolate symbiont Transform symbiont (No Transmission) Reinfect insect with transformed symbiont

Ongoing Studies

• • • •

Acute toxicity in normal and T-cell deficient mice (completed) Chronic toxicity/ allergenicity studies (ongoing) Lab/Greenhouse efficacy studies (ongoing) GGC Proposal

Lymphatic Filariasis (LF)

>120 million infected in over 80 countries Over 1 billion people at risk Second leading cause of disability worldwide High disease burdens: India, Nigeria, Indonesia Mosquito-borne, including vectors of malaria Elimination is a public health goal No vaccine; mass drug administration, hygiene and mosquito control

MF PERIODICITY

Mass Drug Administration – Focus of Control

Effectiveness of DEC Salt in Miton, Haiti

MF in People 25 20 15 10 5 0 Pre-Tx 1 Year Post-Tx % MF Prevalence MF Density (20ul) L3 in Mosquitoes 2.5

2 1.5

1 0.5

0 Pre Tx 1st Qtr 2nd Qtr 3rd Qtr 4th Qtr

Monitor MF in vectors to determine MDA efficacy

Novel Applications of Technology

GC/MS: attractants; surveys based on odors; markers for flight range, e.g., Rb NIR: species ID/ITN status; vector age; counterfeit drugs Acoustics: larval/adult ID; surveillance; infections

Tsetse SIT collaboration – NIRs?

Spectrum of a wheat kernel

700 900 1100 1300

Wavelength (nm)

1500 1700

NIR SORTING Manual ~ 1 pupa per 30 sec Automated ~1 pupa per sec

Sexing Tsetse Pupae by NIRS

• Sex pupae - females returned to the colony • Male pupae irradiated and released • NIRS: ~98% of males and females correctly classified; automated sorting at 1/second

Fly Age Predicted by NIRS

(n=600, R 2 =0.95)

14 12 10 8 6 4 2 0 -2 -2 0 2 4 6 8 Actual Age (d) 10 12 14

Independent of preservation method; better than pterin analysis

The Mosquito Net – by John Singer Sargent