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Project Wolbachia – Singapore Welcomes New Collaborator For Ongoing Phase 2 Field Study

18 Sep 2018

Verily joins Orinno Technology Pte. Ltd. as the second private sector collaborator in the ongoing Project Wolbachia Singapore field study

Singapore, 18 September 2018 – The National Environment Agency’s (NEA) ongoing Project Wolbachia – Singapore Phase 2 field study has been enhanced by the innovative supporting technologies developed by our private sector collaborators. Today, NEA welcomes a new partnership with Verily, the life sciences and healthcare company of Alphabet Inc. from the United States of America, for the ongoing Phase 2 field study. They join local start-up Orinno Technology Pte. Ltd. (OTPL), which had developed process modules that have helped to improve the production efficiency of the Wolbachia-Aedes aegypti mosquitoes required for field releases.

2          Such partnerships bring together advanced engineering technologies and artificial intelligence, that work synergistically in developing novel solutions for Aedes aegypti mosquito population control. Verily's automated mosquito production, sorting and release equipment have been used in successful large-scale field trials with Wolbachia-Aedes aegypti in the US and Australia, producing and releasing millions of male mosquitoes. The new partnership was announced at the opening ceremony of the Fifth Singapore International Dengue Workshop (SIDW), held since 2009 as a joint initiative by Singapore and the World Health Organization (WHO) to provide valuable training on dengue management, surveillance and control, for public health practitioners from all WHO regions worldwide.

Update on Phase 2 Field Study

3          The Phase 1 field study of Project Wolbachia – Singapore, conducted from October 2016 to December 2017 by NEA’s Environmental Health Institute (EHI), saw a 50 per cent suppression of the urban Aedes aegypti mosquito population at the sites where male Wolbachia-Aedes aegypti mosquitoes were released.

4          The ongoing Phase 2 field study, which started in April 2018, aims to improve release methodologies for Singapore’s high-density and high-rise urban landscape to achieve a larger impact and has been progressing well. Interim results show that the additional release of male Wolbachia-Aedes aegypti mosquitoes from high floors at residential blocks at the target sites and at adjoining buffer zones – in comparison to Phase 1 where releases were only made at ground floors at the target sites – have improved the chances of the male Wolbachia-Aedes aegypti mosquitoes mating with the urban female Aedes aegypti mosquitoes in Singapore’s high-rise residential blocks. The release of Wolbachia-Aedes aegypti males at adjoining buffer zones is also helpful in reducing the infiltration of urban males and females into the release sites.

5          This can be seen in the reduced viability of Aedes aegypti eggs collected from the release site at Yishun (eggs from the mating of Wolbachia-Aedes aegypti males with urban Aedes aegypti females are non-viable). During the eight-week period of such releases at the Yishun study site, 90 per cent of Aedes aegypti eggs collected at the release site were non-viable, suggesting that there had been successful mating between the released male Wolbachia-Aedes aegypti mosquitoes and the female urban Aedes aegypti mosquitoes, resulting in non-viable eggs due to biological incompatibility between the released and urban mosquitoes. This is an improvement from the 50 per cent egg viability observed in Phase 1, when male Wolbachia-Aedes aegypti mosquitoes were released only from the ground floor. More releases and monitoring are being carried out to determine if the decline in egg viability is sustainable, and if it is able to lead to a significant suppression of the urban Aedes aegypti mosquito population.

Inter-sectoral Collaboration and New Partners for Project Wolbachia – Singapore

6          As Project Wolbachia – Singapore makes progress, NEA’s Environmental Health Institute (EHI), a centre of excellence for vector biology, welcomes new collaborations with local and international partners to further the development of engineering and artificial intelligence (AI) solutions for Wolbachia-Aedes aegypti technology. Please see Annex A for more information on local and international collaborators for Project Wolbachia – Singapore.

7          One such partnership is with Orinno Technology Pte. Ltd., with which NEA’s EHI is jointly developing automated devices, to increase the productivity, consistency and quality of the Wolbachia-Aedes aegypti mosquitoes produced, thus enabling effective scale-up of production. The larvae counter is able to count mosquito larvae 30 times faster than manual methods, and has significantly reduced the time needed to count 4,000 mosquito larvae, from two hours to three minutes. This effectively enhanced the mass production and sorting of male Wolbachia-Aedes aegypti mosquitoes required for the Phase 2 field study.

8          NEA’s EHI is also collaborating with Verily to test an automated mosquito sex-sorter and a mosquito release device developed by the company. Verily’s adult mosquito sex-sorter uses artificial intelligence to separate the male from the female mosquitoes with very high accuracy, which will help prevent the release of females. It would thus avoid the build-up of female Wolbachia-Aedes aegypti mosquitoes in the field, and ensure that the released male Wolbachia Aedes aegypti mosquitoes remain effective in suppressing the urban Aedes aegypti population. In addition, Verily has developed an automated cart for dispensing male Wolbachia-Aedes aegypti mosquitoes at Singapore’s high-rise residential blocks. The cart will be tested at the Tampines West study site from September 2018 to January 2019, as part of NEA’s Phase 2 field study. More information on the Verily release system used for Project Wolbachia – Singapore can be found in Annex B.

9          “We are excited to partner Singapore’s National Environment Agency to bring Verily’s Debug technology to Project Wolbachia – Singapore. This collaboration will help advance the science of the male Wolbachia-Aedes aegypti method in mosquitoes, and further our understanding of how best to operationalise this technique in dense, urban environments. We hope that by working together, we can see a large reduction in the population of Aedes aegypti, a mosquito responsible for many devastating diseases, at our release sites”, said Mr Linus Upson, Vice-President of Engineering and Head of the Debug Project, Verily.

10        NEA’s EHI has forged a strong network of collaborators, from scientists, mathematicians and engineers in varying fields of expertise, to grassroots leaders and community volunteers, and continues to work closely with its various partners. Michigan State University (USA), provided EHI with the Wolbachia-Aedes aegypti strain. Expert scientists and mathematicians from the University of Melbourne (Australia), Imperial College London (United Kingdom), Saw Swee Hock School of Public Health, National University of Singapore, Genome Institute of Singapore, A*STAR and Duke–NUS Medical School, have also contributed to the project.

Collaboration and Capacity Building in the Fight Against Dengue

11        These collaborations between the different sectors are part of our 3P (People, Public and Private) partnership that is central to Singapore’s dengue control programme, and to our efforts in developing innovative and sustainable initiatives to combat dengue and other Aedes-borne diseases. To tackle this complex dengue problem, many heads are better than one, especially when each brings with it different complementary expertise”, said Mr. Khoo Seow Poh, Deputy Chief Executive Officer of NEA, at the opening address of the Fifth Singapore International Dengue Workshop (SIDW).

12        The partnerships were highlighted at the opening ceremony of the Fifth SIDW, which promotes and trains international public health practitioners on dengue management, surveillance and control. Scheduled to run from 18 to 27 September 2018, SIDW is co-organised by NEA’s Environmental Health Institute, the World Health Organization (WHO), the Ministry of Foreign Affairs and the National Centre for Infectious Diseases, in partnership with Ngee Ann Polytechnic, and supported by the Singapore Society for Microbiology and Biotechnology. SIDW is a unique workshop that offers public health practitioners a holistic view of dengue surveillance and control, with an emphasis on the establishment of linkages between laboratory diagnosis and surveillance, clinical management and field control. Please see Annex C for more information on the Fifth Singapore International Dengue Workshop (SIDW).

13        Formerly known as the Asia-Pacific Dengue Workshop, the previous four workshop runs have trained more than 240 public health practitioners from 41 countries. For this fifth run of the workshop, we welcome 86 participants from 24 WHO Member State countries – representing the Western Pacific, South-East Asia, Eastern Mediterranean and African WHO Regions, including the Pan American Health Organization – comprising vector control programme managers, laboratory specialists, clinicians, researchers and policymakers. Other than sharing current best practices in dengue control, the workshop will also be introducing participants to new technologies such as Wolbachia, Gravitraps for Aedes mosquito surveillance, use of machine learning for outbreak prediction, and use of the Geographical Information System for generating situational awareness and planning of dengue control operations.

14        Collaboration has been key to WHO’s efforts in capacity building and leveraging new technologies to tackle mosquito-borne diseases. Dr. Rabindra Abeyasinghe, Coordinator, Division of Communicable Diseases, Western Pacific Regional Office, WHO, who delivered the opening lecture at the workshop said, “WHO is thankful to the Government of Singapore for its willingness to support other countries, not only in Asia-Pacific but also beyond, to address the challenges posed by dengue, through sharing its experiences. Singapore has achieved unique success in clinical management, surveillance and control through proactive planning, careful attention to detail and a willingness to adopt new technologies. There are useful lessons to be learnt by other countries facing the challenges of dengue, and WHO is pleased to jointly organise this workshop.”

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Collaborations for Mosquito Production and Release Solutions

  • The Environmental Health Institute (EHI), of the National Environment Agency (NEA), has been working with various collaborators to explore and develop engineering and artificial intelligence solutions, to support the production and release of male Wolbachia-Aedes aegypti mosquitoes for Project Wolbachia – Singapore. Mosquito rearing is laborious and time-consuming, with very little technological support, despite advances in engineering and AI solutions in other fields.

Collaboration with Michigan State University (USA) and Sun Yat-sen University-Michigan State University (China)

  • EHI obtained Aedes aegypti eggs carrying a Wolbachia strain from its collaborator –Professor Xi Zhiyong – based at the Department of Microbiology and Molecular Genetics, Michigan State University, US; and Director of Sun Yat-sen University–Michigan State University Joint Center of Vector Control for Tropical Diseases, China. Professor Xi Zhiyong had extracted a Wolbachia strain from the Aedes albopictus mosquito (another black-and-white mosquito species commonly found in Singapore), and introduced the bacterium into Aedes aegypti eggs via microinjection, causing adult mosquitoes that emerged to carry the bacterium. As Wolbachia is passed on from female Wolbachia-Aedes mosquitoes to their offspring, the females produced are used for generating more Wolbachia-Aedes mosquitoes in the laboratory, whilst the males are used for release in the field for Aedes aegypti population control.
  • In March 2015, Professor Xi Zhiyong and his team released 500,000 male Wolbachia-Aedes albopictus mosquitoes (a mosquito species responsible for most of the mosquito-borne diseases in China) at Shazai Island, Guangzhou, China. Results from a year of study showed that the mosquito population on the island had been reduced by 99 per cent.
  • In January 2017, Professor Xi Zhiyong received a USD 1 million grant from the United States Agency for International Development to build a mosquito factory in Yucatan, Mexico. Professor Xi Zhiyong is coordinating with multiple agencies of the Mexican government, the governor of Yucatan and Universidad Autonoma de Yucatan, to breed mosquitoes and conduct field tests in Mexico. Two villages have been selected for release, and community education and baseline data collection are underway.

Collaboration with Orinno Technology Pte. Ltd. (Singapore)

  • EHI and Orinno Technology Pte. Ltd., have come together to develop various novel automated devices that will increase the productivity and quality of the mosquitoes reared.
  • One of the inventions – the larvae counter – has yielded successful results, and has shortened the time required to count 4,000 larvae, from two hours to three minutes. This has allowed EHI to produce enough male mosquitoes for the Phase 1 field study.

Devices developed with Orinno Technology Pte. Ltd.


Innovation developed

Brief Description


Larvae counter

  • Larvae purification and concentration system based on light and micro-fluid control
  • 30 times faster than manual counting methods


Pupae counting and dispensing module

  • Pupae purification and concentration system based on light and fluid control
  • Estimated 20 times faster than manual counting methods


Sex-sorting module


  • High speed image processing to analyse pupae size profile in mass
  • Automated separation (without need for human involvement) of males and females at both pupal and adult stages


Collaboration with Verily (USA)

  • Verily is the life sciences and healthcare company of Alphabet Inc. in the United States. EHI is collaborating with Verily in the Phase 2 field study of Project Wolbachia – Singapore, to test the automated mosquito sex-sorting and release mechanisms developed by Verily. Please see Annex B for details of the mosquito release cart.
  • Verily’s experienced mosquito scientists are collaborating with NEA on mosquito rearing and other aspects of the Phase 2 field study.
  • Verily’s Debug Project has conducted successful large-scale field trials of its automated mosquito rearing, sex-sorting and release systems in both the US and Australia.
  • In July 2017, Verily collaborated with MosquitoMate and the Consolidated Mosquito Abatement District, Fresno County, US, to launch Debug Fresno – a field study that aimed to suppress the Aedes aegypti population in Fresno County, California, US, by releasing male Wolbachia-Aedes aegypti mosquitoes. Automated mass rearing and sex-sorting processes developed by Verily enabled the release of one million insects a week at full production. The use of mobile release devices developed by Verily also allowed mosquitoes to be released in an even and targeted way during the breeding season. Results showed that during the peak of the mosquito season in 2017, there was an average of 68 percent reduction in female Aedes aegypti mosquitoes at the release area relative to other similar sites. Debug Fresno is continuing during the 2018 mosquito season (April to December) with preliminary results showing an even greater reduction in biting, female Aedes aegypti mosquitoes.
  • Using its automated and scalable mosquito rearing and sex-sorting methods, and mosquito release devices, Verily partnered the Commonwealth Scientific and Industrial Research Organisation and James Cook University, Australia, to launch Debug Innisfail in November 2017 – releasing male Wolbachia-carrying Aedes aegypti to reduce the Aedes aegypti population in Innisfail, Australia. The field study concluded in June 2018, and initial results showed that the presence of biting, female mosquitoes at the three towns where releases occurred had decreased by more than 80 per cent over three months.

Collaborations for Study Design, Data Analytics and Modelling, and Biological Mechanisms

Collaboration with University of Melbourne (Australia)

  • EHI is collaborating with Professor Ary Hoffmann, at the University of Melbourne, Australia, on study design, data analytics and modelling for Project Wolbachia – Singapore.
  • Professor Ary Hoffmann (Australian Laureate Fellow and Professor, School of BioSciences and Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Australia) is also a member of NEA’s Dengue Expert Advisory Panel (DEAP).
  • DEAP was appointed by NEA in June 2014 to provide professional advice on new modalities for dengue control, particularly on the use of Wolbachia for suppression of the Aedes aegypti mosquito population. The roles of the Panel are to:

a. Review latest scientific literature on dengue infection, with particular emphasis on risk factors and identifying new modalities for disease control;

b. Advise NEA on the planning, experimental design and implementation of appropriate interventional studies;

c. Assess the suitability of using male Wolbachia-Aedes mosquitoes for reducing the impact of dengue in Singapore, taking local epidemiology and ecology into consideration; and

d. Review field study/ trial and results.

Collaboration with Imperial College London (United Kingdom)

  • EHI is collaborating with Professor Neil Ferguson, at Imperial College London, United Kingdom (UK), on data analytics and modelling for Project Wolbachia – Singapore. The collaboration aims to develop a mechanistic model for mosquito population, to assess the impact of male Wolbachia-Aedes releases on the mosquito reproductive number.
  • Professor Neil Ferguson (Founding Director of the Medical Research Council Centre for Outbreak Analysis and Modelling at Imperial College London, and Director of the Health Protection Research Unit for Modelling Methodology under the UK National Institute for Health Research) is also a member of NEA’s DEAP.

Collaboration with Saw Swee Hock School of Public Health, National University of Singapore (Singapore)

  • EHI is collaborating with Associate Professor Alex Cook from Saw Swee Hock School of Public Health, National University of Singapore, on data analytics and modelling for Project Wolbachia – Singapore. As NEA progresses towards a randomised trial for impact assessment of Wolbachia technology and in order to determine the overall effectiveness of the technology, the collaboration will seek to design a robust study, conduct data analysis, develop data visualisation tools, evaluate the impact of Wolbachia and other intervention methods, and conduct cost-benefit analysis of the Wolbachia approach.

Collaboration with Duke–NUS Medical School (Singapore)

  • EHI is collaborating with Assistant Professor Julien Pompon from Duke–NUS Medical School to gain in-depth understanding of the mechanisms of cytoplasmic incompatibility (CI), induced by different strains of X-ray treated and non-treated male Wolbachia-Aedes and urban female Aedes aegypti mosquitoes. This is part of NEA’s mitigation measures for the potential risk of failure, to pre-empt any potential loss of function of CI, or the potential development of resistance of Singapore’s urban Aedes aegypti mosquitoes to CI. Furthermore, a better characterisation of CI will improve use of the Wolbachia suppression strategy in Singapore.

Collaboration with Genome Institute of Singapore (Singapore)

  • EHI is working with Dr. Paola Florez De Sessions from the Genome Institute of Singapore on genomic characterisation of vector-borne pathogens and vector populations by Next Generation Sequencing, to understand the genetic structure of vector populations that may contribute to vector-borne disease transmission in Singapore, which is a component that forms the risk assessment framework of the Wolbachia-Aedes initiative. The success of the Wolbachia-Aedes approach depends on the ability of Wolbachia-Aedes aegypti mosquitoes to compete with the urban Aedes aegypti population and express the desired phenotypes, regardless of Aedes aegypti genetic variations. There is thus a need to regularly monitor the proportion of Wolbachia-Aedes aegypti mosquitoes in the field, as well as the temporal and spatial shifts of genetic diversity of the urban Aedes aegypti population in Singapore.



How the Release Device Works

  • The automated release device is able to dispense male Wolbachia-Aedes aegypti mosquitoes at the ground and high floors of residential blocks to achieve even, controlled releases.
  • The automated release device works in the following manner:

a. A release cart, operated by one person, is pushed along the corridors of residential blocks.

b. Bluetooth beacons (small coin-sized devices that respond with a unique signal when triggered by a transmitter) will be installed beforehand at corridors of residential blocks to help reliably trigger the release of batches of male Wolbachia-Aedes aegypti mosquitoes.

c. The male Wolbachia-Aedes aegypti mosquitoes are gently blown out from the nozzle of the cart in controlled doses evenly along the corridor, outwards away from residential units and above the height of the balcony balustrade.

Wolbachia 18 Sep

Fig. 1. Infographic showing the release system: cart and beacon (not to the same scale: beacon is coin-sized)



About the Singapore International Dengue Workshop

  • The Singapore International Dengue Workshop (formerly known as the Asia-Pacific Dengue Workshop) is a capacity and capability building programme for knowledge sharing and exchange, which supports the Western Pacific Regional Action Plan for Prevention and Control of Dengue (2016), endorsed by Western Pacific Member States and the WHO Global Strategy for Dengue Prevention and Control 2012-2020.

    The workshop is jointly organised by the Environmental Health Institute (EHI) [a World Health Organization Collaborating Centre for Reference and Research of Arbovirus and their Associated Vectors, and a centre of excellence for vector biology] of the National Environment Agency (NEA), the World Health Organization (WHO), the Ministry of Foreign Affairs (MFA) under the Singapore Cooperation Programme, and the National Centre for Infectious Diseases (NCID), in partnership with Ngee Ann Polytechnic (NP), and supported by the Singapore Society for Microbiology and Biotechnology.

Workshop Objectives

  • The aim of the workshop is to enable the participants:
    • To be trained in best practices for dengue surveillance, control and management
    • To build capacity and acquire relevant knowledge and skills in implementing and strengthening key components of the national integrated dengue control programmes
    • To share clinical management guidelines and best practices for dengue
    • To promote close inter-sectoral collaboration amongst laboratory investigation, field operations, and clinical management and surveillance
    • To encourage a dengue laboratory surveillance network within the region
    • To promote regional and global dengue surveillance efforts in alignment with the Asia-Pacific Strategy for Emerging Diseases and Public Health Emergencies (APSED III), the Western Pacific Regional Action Plan for Prevention and Control of Dengue (2016), and the WHO Global Strategy for Dengue Prevention and Control 2012-2020
  • This year, besides local experts and lecturers from NEA, MOH, NCID, NUS, NTU and NP, there will also be invited expert speakers from the following institutions/ organisations:



Institutions/ Organisations:



World Health Organization (WHO)


  • WHO Headquarters (Geneva)
  • Western Pacific region
  • Eastern Mediterranean region



  • Chinese Centre for Disease Control and Prevention (National Institute for Communicable Disease Control and Prevention)
  • Yunnan Institute of Parasitic Diseases



  • Ministry of Health (Disease Control Division)
  • Universiti Sains Malaysia (School of Biological Sciences)



  • Norwegian University of Life Sciences (visiting at Kasetsart University, Thailand)


Sri Lanka

  • Ministry of Health (National Dengue Control Unit)

Uniqueness of the Singapore International Dengue Workshop Series

  • It is a unique workshop that offers public health practitioners a holistic view of dengue surveillance and control, with an emphasis on the establishment of linkages between laboratory diagnosis and surveillance, clinical management and field control.
  • The curriculum is specially designed to meet the training needs of public health practitioners responsible for:
    • Laboratory diagnosis and surveillance
    • Vector surveillance and control
    • Clinical management
  • A technical panel comprising representatives from WHO, Singapore’s NEA and NCID, and Malaysia’s Ministry of Health (MOH), has been convened to review the curriculum to ensure that it continues to be relevant to the needs of public health practitioners in the region and beyond.

Outcomes of Previous Runs

  • To-date, more than 240 public health practitioners from 41 countries have benefited from the Dengue Workshop series, and have been trained over the four workshop runs held in 2009, 2010, 2012 and 2014.
  • The first workshop in 2009 started with countries from the WHO Western Pacific and South-East Asia regions. By the fourth workshop, we had additionally welcomed participants from the Eastern Mediterranean and African regions, as well as practitioners from non-endemic countries – showing the increasing threat of dengue globally.
  • Evaluation of post-workshop surveys to gauge the impact of the workshop – gathered from alumni of the workshops – revealed the very significant gain in knowledge and skills; gain in confidence in executing their work; and application of the knowledge to their work back in their home countries. For example, past participants from the Solomon Islands have now established a new PCR laboratory for dengue confirmation; participants from the Philippines have initiated the use of rapid diagnostic test kits; and new measures have been introduced in Fiji, Indonesia, Malaysia and Micronesia to further enhance their respective dengue control programmes.
  • The participants also greatly benefited from the broad-based curriculum and cross-disciplinary integrated approach, which offered them a comprehensive perspective of dengue prevention and control, ranging from clinical management, to laboratory diagnosis and field vector control.
  • The previous workshops also generated some positive outcomes and spin-offs:
  • UNITEDengue (UNited In Tackling Epidemic Dengue) regional surveillance

➢ UNITEDengue is a cross-border partnership to achieve shared goals in dengue control, with the following objectives:

(i) Cross-border case and virological dengue surveillance to allow timely sensing of the dengue situation in the Asia-Pacific region;
(ii) Regional capacity building and strengthening for dengue viral and entomological surveillance; and
(iii) Regional capacity building and strengthening for dengue control.

➢ UNITEDengue is an outcome of the highly successful first two Dengue Workshops (2009 and 2010), and was officially launched at the third workshop (2012). Since its official launch by the two founding partners (Singapore and Malaysia) in September 2012, the network has now expanded to 11 countries. Additional partners are: Brunei, Cambodia, Lao PDR, Myanmar, Pakistan, Philippines, Sri Lanka, Thailand, Vietnam.

➢ UNITEDengue has received the support and concurrence of the ASEAN Health Sector, as the platform for sharing of dengue data in the region.

➢ A joint publication with Malaysia Ministry of Health entitled ‘2013 Dengue Outbreaks in Singapore and Malaysia Caused by Different Viral Strains’ was published in the American Journal of Tropical Medicine and Hygiene (June 2015). EHI also provides regular reports following analysis of available data on the dengue situation.

  • Identification of a critical need to sustain regional expertise in taxonomy for mosquitoes of public health importance

➢ To address this gap, in partnership with the Walter Reed Biosystematics Unit of the Smithsonian Institution in the United States, an Advanced Mosquito Taxonomy Workshop was held in 2011 to build up the capability for mosquito species identification in the surveillance of mosquito-borne diseases.

  • Set-up of an informal social media network for Dengue Workshop alumni, to share dengue updates and to expand and sustain partnerships

➢ Initiated by participants of the third Dengue Workshop, the social media group allows participants to stay in touch with each other, as well as share dengue updates easily. Through this network, past participants have also been able to provide feedback and comments about the workshop and its benefits to their work.