ECOLOGY & GENETICS OF Anopheles gambiae ON ISLANDS

Bijago Islands Guinea- Ecology and Genetics of Anopheles gambiae on Islands 05/08/2009 - 04/30/2011

NIH Grant Number: R21AI062929

Progress toward the development of genetically modified mosquito (GMM) strains to be used in a genetic control program for mosquito vectors of malaria has reached a point where most of the scientific community involved in this research are beginning to think about field trials. The following recommendation was made by a group of experts in the field of vector biology participating in a meeting entitled "Genetically Engineered Arthropod Vectors of Human Infectious Diseases": "The exciting progress in molecular strategies for blocking parasite development in vectors has far outpaced the related and essential work on field sites and vector populations that must precede any GM release field trial." Imperial College, London 14-15 September, 2001.

We believe that the most promising sites for GMM field trial sites would be on islands off the coast of sub- Saharan Africa. With this Exploratory/ Developmental Research Grant (R21) we initiated studies aimed at assessing the suitability of two island sites. We selected one island off the east coast of Africa and one off the west to explore for this program. Based on a critical analysis of available data we chose to study Anopheles gambiae populations on Ihla de , part of the Bijago Archipelago in West Africa and Grande Comore in East Africa. Our initial assessment included:

(1)collection of basic ecological and biological data for An. gambiae at each site, including relative population densities, adult and larval distributions;

(2)an assessment of the degree to which island populations are genetically isolated from the nearest mainland populations and from populations on neighboring islands, and a description of the genetic structure of populations on each of the two target islands. ISLANDS MALARIA PROJECT

The Bijago Islands, Guinea-Bissau PARTNERSHIP

Instituto Nacional de Saúde Pública

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University of California-Davis GUINEA-BISSAU PROJECT WORKPLAN OCTOBER - NOVEMBER 2009

I. Mainland collections:

1) Antula ~ 5 km north of Bissau 2) Prábis 3) Mansoa 4) Bambadinca ~ 75 km east of Bissau 5) Enxudé near Bissau, south of the Rio Gêba (there is a ferry that goes from Bissau to Enxudé) 6) Fulacunda

II. - Island collections:

1) Ilha de - Bolama 2) Ilha de - Inorei 3) Ilha de Orango - Eticoga 4) Ilha de - Abu

Contact Info for Guinea-Bissau Personnel:

Jao Dinis: [email protected] Daniel Otiudi Go: [email protected] Bijago Island Expedition, October-November 2009. l. to r. Jao Denis, Yoosook Lee, Anton Cornel, Greg Lanzaro, George ?, Chuck Taylor, Daniel Otiudi Go. Mosquito Collection sites

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1. Bissau / Antula 5. Prabis 2. Bruce, Ilha do 6. Gabú 3. Eticoga, Ilha do Orango 7. Canjufa 4. Abu, Ilha do Formosa 8. Bambadinca PHOTOLOG Bijago Islands Project October/November 2009 Greg Lanzaro, Anton Cornel, Chuck Taylor & Yoosook Lee Monday, October 26, 2009 –

Arrive Bissau at 1:30 AM, spent night at Solmar Hotel, Bissau. Met by Jao Dinis. Met with Cesareo Martins at the hotel later that day to arrange for transportation to Bubaque Island and have dinner.

Hallway outside rooms at the Solmar Hotel Front of Solmar Hotel

The registration desk in the lobby of the Solmar Hotel At dock in Bissau arranging transport to the islands. Cesaro (foreground), Chuck, Yoosook Tuesday, October 27, 2009

Departed for Ilha do Bubaque in small boat with the pilot, George, Jao and assistant, Daniel about two hours by boat to Bubaque and checked into the Casa Paulino Hotel in the main town of Bubaque.

Our transportation from Bissau to the Bijagos Islands, Yoosook and Jao on the way to Bubaque the small boat, which turned out to be wholly inadequate.

Greg and Chuck on trip to Bubaque Abandoned hotel in the town of Bubaque. Tuesday, October 27, 2009 Ilha Bubaque

Casa Paulino Hotel , Bubaque. Down the street from hotel.

Street in Bubaque. Coast of Bubaque Island. Wednesday, October 28, 2009

Drove by ambulance to the village of Bruce, met with the chief and attempted to collect adults, but could not find any.

………………………………………….This ambulance was one of few vehicles on the island………………………………………………..

……………………………………………………………….…….….village of Bruce………………………………………………………………………… Wednesday, October 28, 2009

Found a couple of flooded pools in the road outside the village of Bruce that contained larvae so we made a good larval collection.

Yoosook, Jao and Daniel collecting larvae. Yoosook, Anton and Daniel collecting larvae.

Chuck and his following. Collecting larvae with trusty ambulance in background. Thursday, October 29, 2009

Collected larvae from site near hospital, on Bubaque. Friday, October 30, 2009

Departed 8:30 AM for Orango Island. Arrived at Orango Parque Hotel and checked in.

Arriving on Orango Island. There was no dock so we Patio and reception desk. had to wade ashore with our gear.

Path from hotel down to the beach. Fisherman near our hotel. Friday, October 30, 2009

Ilhao de Orango, Orango Parque Hotel. Friday, October 30, 2009

Walked from hotel to village of Eticoga, about 2km.

Hiking from hotel to the village of Eticoga. Road to Eticoga, on right.

Chatting with a man on the road. Little girl in Eticoga Friday, October 30, 2009

At Eticoga met with clinic nurse, Augusto da Lama, then with the chief, Augusto Fernando da Pina. He gave us permission to collect the following morning.

Scene in Eticoga. Young girl outside one of the houses.

Chuck on the beach outside Eticoga. Health clinic in Eticoga. Saturday, October 31, 2009

Walked back to Eticoga to collect adults, but found few. Found a flooded pasture with larvae. Yoosook and Greg returned later that afternoon and collected larvae with Jao and Daniel. Anton and Chuck returned to Eticoga to try to do adult collections with Jao and Daniel that night, but were unsuccessful. Anton, Chuck and Greg collected a reasonable number of adults biting at the hotel. None developed ovaries for karyotyping. These were Anopheles gambiae and it turned out that 18% were infected with P. falciparum.

In the village of Eticoga. Preparing collecting equipment.

Collecting larvae in a flooded pasture just outside the village. Looking for adults in the same pasture. Sunday, November 1, 2009

Took day to see salt-water hippos and did night catch again at hotel.

Hiking through saw grass in search of hippos. Group (called a ‘crash’) of salt-water hippos in a small pond not far from the hotel. Big bull in center of photo became a bit aggressive.

We picked up a lot of large, black leeches crossing this stream Beach near our hotel. Monday, November 2, 2009

Traveled in heavy rain to Ilha de Formosa to meet with village nurse at village of Abu. Collected good number of adults

Boat ready to depart Ilha de Orango for Ilha de Formosa. Arriving on Ilha de Formosa.

Home and office of the village nurse. Heading out in search of mosquitoes. Tuesday, November 3, 2009

Spent day at Abu, Ilha de Formosa collected good number of adults. Trip back to Bubaque was very bad in choppy waters.

Outside the village of Abu. House on California Street (Rua Califonia) in Abu.

Trail outside of Abu. Return to nurses house after collecting. Wednesday, November 4, 2009

Traveled from Bubaque to Formosa for additional collection of adults from Abu. Collected for about one hour and then went on to Bissau. Terrible trip, rough waters everyone badly shaken by pounding of boat. Checked back into Solmar Hotel in Bissau.

Our trusty transportation while on Bubaque. Room at the Solmar Hotel, Bissau.

Roman Catholic cathedral in Bissau Relaxing at café in Bissau. Thursday & Friday, November 5-6, 2009

Collection trip to Antula, about 5 km outside Bissau and Prabis, about 25 km SW of Bissau. On Friday Greg stayed back in Bissau with Chuck because of stomach problems. We made a large collections of adults at both sites. Chuck departs for return to States.

Hiking from main road to the village of Prabis. House in the village of Prabis.

Children in the village of Prabis. Moistening towel to keep adult mosquitoes humidified. Saturday & Sunday, November 7-8, 2009

Collection trip to Gabú, about 200km northeast of Bissau. Checked into hotel and traveled to village of Canjufa, ~25 km northeast of Gabú. Were unable to make a good adult collection (N=12), so we collected larvae along the road between Canjufa and Gabú. Were able to collect 50 larvae from 3 road pools. On Sunday we stopped to collect in village of Bambadinca, about75 km northeast of Bissau. Made a collection of adults (N=50). After collecting we returned to the Sol Mar Hotel in Bissau.

………………………………………………………Collecting larvae in roadside pool between Canjufa and Gabú……………..…………………………………….. Monday, November 9

Met Amabilia in the morning at the National Institute of Public Health office to review our work. She expressed an interest in further collaboration. After the meeting we returned to Prabis to make a collection of adults. Made a large collection of adults which Anton and Greg put into two one gallon cages to hold for bloodmeal digestion and oviposition.

At the National Institute of Public Health office in Bissau. At the National Institute of Public Health office in Bissau. Left to right; Yoosook Lee, Amabelia Rodrigues, Anton Cornel Cesario Martins, Greg Lanzaro and Jao David. Tuesday, November 10, 2009

Returned to Antula, near Bissau to make an additional collection of adults. Want a large collection from here since this is the site from which high frequencies of M/S hybrids were high. We departed Bissau on Thursday, November 12 traveled home via Lisbon and Amsterdam.

Anton using mouth aspirator to collect resting Yoosook searching for adult A. gambiae in house. adults in a house in Antula.

Back at hotel in Bissau sorting and storing Yoosook preparing labels and placing specimens in tubes. mosquitoes collected in Antula. Jao, Anton & Yoosook. Publications

2011. 6(11): e27920. doi:10.1371/journal.pone.0027920

2013. doi:10.1111/eva.12056