France Daigle of the University of Montreal in Canada will identify the microorganisms that enable the survival of the typhoid fever-causing bacterium, Salmonella enterica serovar Typhi, at low levels in water, and thereby enhances disease spread. Typhoid fever spreads through contaminated food and water, and results in over 125,000 deaths annually worldwide. S. Typhi are so-called auxotrophic bacteria because they rely on an external source of the essential amino acids that they need to grow. Microbial interactions may provide nutrients and also increase bacterial fitness and support persistence by protecting them from the environment, thereby increasing the rate of disease transmission. They will assemble a microbial community in water consisting of three components: one protozoan (from a group known to promote bacterial survival); a defined consortium of bacteria representative of the human fecal microbiota; and fluorescently-tagged S. Typhi. They will evaluate the ability of S. Typhi to grow in these microcosms, and how they grow, such as in biofilms or inside the protozoa. They will also determine whether these persistent S. Typhi are better able to infect and survive in human cells. Finally, water samples from an endemic region in East Africa will be analyzed for the presence of S. Typhi and identified beneficial microbial partners using quantitative PCR.

Matthew DeLisa of Cornell University in the U.S. will create a cell-free synthetic biology platform for low-income settings that produces thermostable polysaccharide-based conjugate vaccines against diarrheal pathogens upon the addition of water to a single tube. Half-a-million children under age five die each year from diarrhea and dysentery, the majority in low- and middle-income countries. Two major causes of bacterial diarrhea are enterotoxigenic E. coli (ETEC) and Shigella strains. Conjugate vaccines combine multiple antigens into one vaccine to increase its activity. However, they require a complex manufacturing process, living cells, and refrigerated storage, which limit their application in developing countries. They will develop the materials and methods for manufacturing thermostable anti-diarrheal vaccines in single tubes that only require the addition of water just ahead of administration. The tubes will contain a plasmid that can express an FDA-approved carrier protein, along with selected O-antigen-polysaccharides from ETEC or Shigella strains, and an enzyme that can conjugate the two via glycosylation, all within a freeze-dried pellet. Following development, they will test the safety, scalability and portability of the vaccines, and characterize their ability to generate effective antibodies that can kill the bacteria. The system is expected to reduce conjugate vaccine costs, and its modular nature will facilitate expansion to other vaccine-preventable diseases.

Robert Kingsley of the Quadram Institute Bioscience in the United Kingdom will locate the typhoid fever-causing bacteria S. Typhi in water reservoirs in Harare, Zimbabwe, and identify any associated protozoa species present in the water that may be supporting disease spread. Typhoid fever is endemic in Zimbabwe, with several major outbreaks reported in the last decade. The bacteria persist in unclean aquatic environments, possibly supported by protozoa, and are transmitted to humans through ingestion of contaminated drinking water. They will detect S. Typhi in sewage effluent and low-quality drinking water in hotspots of typhoid transmission by enrichment culture and PCR, and use whole genome sequencing to establish the phylogenetic relationship between these bacteria and clinical typhoid isolates in the same city. They will also amplify 18S rDNA from the sewage and drinking water samples to characterize the microbial community in water and define the protozoa population. These data will help identify potential synergistic interactions between S. Typhi and other microbes to inform prevention strategies.

Hasan Uludag of RJH Biosciences in Canada will develop an affordable immunotherapy system based on genome-integrating transposons that works inside the body for the treatment of a wide variety of diseases such as cancer and diabetes. Emerging immunotherapies offer promising treatment for many diseases, but they require genetic modification of immune cells outside the body, and are thus labor intensive and expensive, limiting their utility in developing countries. They will use engineered nanoparticles in a new approach to immunotherapy that modifies immune cells inside the body. The nanoparticles are derived from polymeric materials that can encapsulate nucleic acids and proteins and release them into host cells. These nanoparticles will be dispersed in a hydrogel matrix with immunostimulatory molecules to create a living bioreactor inside the host that will attract and genetically modify immune cells. They will select polymers for their ability to deliver DNA-based transposons (to facilitate integration into the host genome) to immune cells and to stably express a reporter gene. Optimal polymers will be transferred into mice and they will evaluate transfection efficiency into immune cells with a fluorescent reporter gene. Finally, they will test the therapeutic efficacy of their in situ immune cell engineering approach in a mouse leukemia model.

Tovi Lehmann of the National Institute of Health in the U.S. will establish cross-country networks of aerial sampling stations in Africa to monitor windborne movement of insects and pests, and evaluate risks to public health, food safety, and ecosystem stability. Vector-borne disease is among Africa's top health priorities, and control of the insect vectors is the primary target for prevention. They will use a unique aerial sampling program to collect airborne insects across Mali and Ghana, and identify insects and pathogens within them by molecular analysis. Sticky nets mounted on helium balloons have shown, in a pilot project, to collect diverse samples, more representative of area fauna than ground sampling protocols. The same project showed that mosquitoes frequently travel (and may spread disease) over hundreds of kilometers. Overnight aerial sampling will be conducted ten nights per month for six months, followed by insect taxonomic identification and RNA/DNA sequencing to identify insects and pathogens. Weather data will be collected from the sampling stations at both ground level and sampling altitude and combined with population data for statistical analysis and simulation of flight patterns. They will produce dynamic, species-specific maps of select insects and pathogens with putative sites of origin, routes and destinations, which will be used to evaluate risks to public health and food security.

Windy Tanner, formerly at the University of Utah and now at Yale University in the U.S., together with Jim VanDerslice of the University of Utah and colleagues from Mehran University of Engineering and Technology in Pakistan, will analyze water samples to determine the conditions that promote the survival of the typhoid fever causing bacterium Salmonella Typhi, and they will use metagenomic deconvolution to identify any gene exchange from other microbial species that may produce drug-resistant strains. S. Typhi is responsible for over 100,000 deaths each year, mostly in the developing world where fecal contamination of food and drinking water is common. The emergence of drug-resistant strains has limited the available treatment options. Biofilms are environmental niches with complex microbial communities and are ubiquitous in the environments where S. Typhi is commonly found. They will sample water and biofilms from a variety of these environments along the fecal-drinking water transmission route in the Sindh province of Pakistan and test for the presence of S. Typhi using qPCR and culture methods. They will also evaluate whether specific organisms stabilize and protect S. Typhi in these biofilms and could cause resistance gene exchange.

Andrew Greenhill of Federation University Australia in Australia, along with partners at the Papua New Guinea Institute of Medical Research, will use advanced environmental microbiology methods to study microbial community dynamics associated with survival of the typhoid fever-causing bacterium Salmonella Typhi in aquatic environments in Papua New Guinea. "Typhoid Mary" Mallon was an Irish-American cook, written into infectious disease folklore as the first asymptomatic carrier of S. Typhi. More than eighty years after her death, little is still known about how the bacteria persists in environmental niches such as contaminated water, which is a major route of disease transmission. They will collect and filter water samples from streams in areas where the disease is common over an eight-month period covering both wet and dry seasons and analyze the microbial communities within by qPCR. This will be combined with physiochemical parameters of the water (temperature, stream height, photosynthetic activity) collected using DIMPP - a low-cost, late-stage prototype suitable for use in low-resource environments - to build network models. These models will be used to identify mathematical connections between environmental factors, aquatic populations, and the presence of antibiotic resistance genes. These connections could be used to develop an early warning system for impending outbreaks.

Feng Fu of Dartmouth College in the U.S. will use social networks to promote positive attitudes and overcome negative views of vaccinations and thereby increase demand. The success of vaccinations has led to steep declines in the incidence of many serious diseases. However, this has decreased the perception of disease risk and thereby lowered vaccination coverage as parents concerns switch to other factors, such as cost and the perceived risk of the vaccination itself, which are fueled via social media channels. These current low vaccination rates exhibit so-called hysteresis whereby the past concerns about safety or necessity prevent the rates from increasing even when the concerns have been disproven. To overcome this, they will use computer modeling approaches to test the ability of targeting social networks to leverage social "contagion" (i.e., spread) of positive attitudes to vaccine knowledge. They will use a healthcare intervention dataset from a network of rural villages in Honduras to model how one or more health-related behaviors or beliefs of an individual affects the group to simulate the social contagion process related to vaccines. They will also evaluate the potential positive impact of influential individuals who publicly support vaccination. The results will be used to develop social network targeting algorithms to increase the demand for vaccination. Their modeling results will be validated using the real data from the village networks.

Caroline Aura from the University of Nairobi in Kenya will teach frontline health workers and caregivers new skills so they can apply simple techniques such as swaddling and rocking to lessen the pain and distress of infants during injections to improve vaccination rates. Vaccination rates are still too low in many low-resource settings, which may be due in part to the discomfort they cause infants. This in turn makes caregivers reluctant to obtain all the recommended vaccinations for their children. Methods exist to reduce the associated pain of injections, but health workers lack the knowledge and skills to implement them. To test their approach, they will recruit vaccinators and community health workers at four rural immunization centers and use seminars and workshops to teach them pain-relieving techniques, including using specific positions and making soothing sounds. They will also develop audio-visual training tools and illustrative guides to help teach the techniques to parents for them to use at home as well. All healthy children under 12 months old visiting the centers for a vaccination will also receive one of the pain relief techniques. They will evaluate the ability of the health workers to manage pain, the level of distress of the infants, and the experience of the caregivers.

Anita Shet of Johns Hopkins School of Public Health in the U.S. will seek to increase childhood vaccination coverage in India and Nigeria by identifying opportunities for catch-up vaccinations when under-vaccinated children are hospitalized. Of the three million people who die each year from vaccine-preventable diseases, about half are children under the age of five, many of whom live in areas where vaccinations are available, but inequitably distributed. Inadequately vaccinated children frequently become sick and are hospitalized, yet most leave the hospital without receiving catch-up vaccines because of physical and policy barriers, or perceived contraindications. They will engage stakeholders, including hospital policy makers and community workers to launch MOVE (Missed Opportunities for Vaccine Equity) to identify and correct missed opportunities for vaccination by using child hospital visits to provide vaccine education and access. MOVE has three components: inpatient in-reach, where MOVE staff check immunization records and inform hospital personnel of missing vaccines; immunization service linkage to inpatient care, which ensures that vaccines are available and provides a vaccination schedule at discharge; and community outreach to provide follow-up reminders and education. They will modify an immunization application and reminder tool to record the data and maintain a cloud-based registry, and test their approach in two large community hospitals in India and Nigeria.

Tapash Roy, Subhash Chandir and Toufiq Rahman of IRD Global in Singapore will improve child immunization coverage among the homeless, or floating, populations in urban slums in Bangladesh by offering vaccines during evening sessions at existing shelters. The mortality rate for children under five in Bangladesh is high, and many deaths would be preventable with better vaccination coverage. The lack of an integrated public healthcare delivery system in urban areas has resulted in a large inequity between the rich and poor, and the vaccination rate among the floating population of approximately 25,000 is well below the national average. They will develop and test a plan to provide vaccination services using an unconventional gathering place of the floating populations (called Pavement Dweller Centers) during convenient hours. PDCs have been established on city property to provide temporary shelter, safe drinking water, and sanitation services to the homeless. They will use this infrastructure to provide free, mobile phone-based digital immunization registry vaccinations during extended hour clinics with evening shifts, so visitors don't need to miss work to spend time traveling to a healthcare center. The program will be executed by personnel trained in communication to pre-empt any apprehensions by caregivers, and will involve educating visitors on the importance of vaccination. They will evaluate the feasibility and cost-effectiveness of the program in three PDCs in Dhaka and assess community response by a post-intervention survey.

Andrew Seal of the Institute for Global Health and Development in the United Kingdom will test whether traditional female social groups in Somalia can adopt a participatory learning and action (PLA) approach to improve vaccine knowledge and coverage in humanitarian settings. Vaccine-preventable diseases are prevalent in Somalia; measles is the leading cause of death in children under five, yet less than 40% of children are immunized. This is due in part to lack of knowledge about the benefits of vaccination. The PLA approach is based on the idea that sustainable social change is possible if teachers and learners engage in meaningful dialogue and share ideas and experiences. Abbay-Abbay groups, common throughout Somalia, are social groups of 10-20 women, led by an elected Khalifada (lead woman). They meet regularly and have a core interest in the challenges of child rearing, with most women having direct or indirect experience with losing a child to measles. They will recruit coordinators to support Abbay-Abbay leaders, providing information and facilitating learning around vaccinations. They will evaluate their approach for improving attitudes to vaccination and reducing the incidence of measles via a randomized cluster study.

Joseph Tucker of the London School of Hygiene and Tropical Medicine in the United Kingdom will hold a national crowdsourcing contest to develop a social media-based intervention to improve confidence in childhood vaccines and boost coverage in China. Expert-driven strategies have been launched to promote vaccination coverage in China, but have had limited effect. As an alternative approach, they will apply crowdsourcing to tap into the knowledge of individuals to design a more effective, online intervention. They will open the contest with a call for new ideas that use text, images, and videos to promote vaccinations; enable online evaluation of those ideas by crowd and expert judges; and assemble a steering committee of health experts to produce the finalists. The final content of the intervention will be developed by the finalists in an intensive 'designathon' event. They will test the new intervention in select community health centers in three cities in China and analyze its ability to improve confidence in vaccinations.

Itoro Ata of the Solina Center for International Development and Research in Nigeria will implement a program to support unemployed mothers in Nigeria by linking education on the importance of vaccinations with vocational skills education and training to improve immunization coverage. Many rural areas in Nigeria have consistently low rates of routine immunization and large populations of unemployed, stay-at-home mothers whose children are at risk of vaccine-preventable disease. They will create a direct link between community health systems and economic empowerment programs focused on vocational skill development by creating the Routine Immunization Buddy System (RIBS). Mothers in the program will be placed in small support groups and paired with another woman within the group. The lead woman of each group will be trained by community health workers to provide practical information on vaccines using a teaching tool called Hannun Rigakafi (the immunization hand). Women in the RIBS groups will also be taught about possible work options, such as farming, and offered associated tools and training. Pairing education on vaccines with vocational training should help boost the confidence of the mothers and better motivate them to complete the five routine childhood immunizations for their children. The program will be piloted in the Kaduna state of Nigeria and evaluated for improving vaccination knowledge and demand.

Marnie Winter and Benjamin Thierry from the University of South Australia, together with Tina Bianco-Miotto, Claire Roberts, and Clare Whitehead of the University of Adelaide in Australia and the University of Toronto in Canada, will develop and test short-interfering RNAs (siRNA) high-density lipoprotein (HDL) nanocarriers for the treatment of preeclampsia. Globally, ten million women develop preeclampsia during pregnancy each year, which results in the deaths of 76,000 women and 500,000 babies; 99% of these are in developing countries. Most current treatments focus on treating the symptoms (high blood pressure and proteinuria) rather than the molecular causes. Some of the causative molecules, such as the angiogenesis inhibitor sFlt1, can be blocked by specific siRNAs, but the challenge is targeting the siRNAs to the right cells in the body. HDL delivery systems for this purpose are effective and safe, and both siRNAs and HDLs are stable at room temperature, important for therapies in resource-poor areas. They will optimize the formulation of their HDL nanocarrier manufacturing platform, and characterize siRNA loading, carrier stability, size, cellular uptake, and silencing ability in 2D culture. Further, they will bioengineer an ex-vivo placenta model that fully recapitulates the structural and phenotypic complexity of a preeclamptic placenta and use it to evaluate tissue penetration and silencing abilities of the siRNA-nanocarrier complex.

Martin Karplus of Harvard University in the U.S. will integrate informatics and artificial intelligence approaches to design stable, synthetic antigens based on the viral hemagglutinin (HA) protein to be used as a universal influenza vaccine. Seasonal influenza causes substantial morbidity worldwide, and the development of a universal vaccine is a global health priority. The current HA-based vaccines do not provide broad coverage against multiple strains, and must be administered annually because of the high mutability of the virus. To address this, they will use a new approach to develop vaccines that elicit broadly-neutralizing antibodies effective against many different influenza strains, including avian, swine, and human varieties. First, they will assemble the publicly-available DNA sequences of the HA gene of avian, swine, and human influenza A strains, and use homology modeling with existing protein structures as templates to create a library of antigens that together are predicted to both maintain the immune system’s memory of influenza while enabling a rapid immune response to future seasonal or pandemic strains. The antigen library will be optimized by performing affinity maturation simulations initiated from known germline precursors of broadly-neutralizing antibodies, and combining the results with machine learning, to predict which antigen cocktails will produce antibodies with the greatest breadth of reactivity. The most promising of these immunization cocktails will be efficacy-tested in small animal models.

Peter Kwong of the Vaccine Research Center, National Institute of Allergy and Infectious Diseases in the U.S. will use an informatics-based approach to identify influenza virus epitopes that are especially suited to induce a strong and broad immune response, being conserved, accessible, and with a specific structural flexibility, and develop them as vaccine targets. Influenza is highly contagious and can cause severe illness, particularly among the young, elderly, and those with pre-existing conditions. Current vaccines are only partially effective, protect against single strains, and are generally ineffective against pandemic strains. Starting with three exposed influenza viral proteins, hemagglutinin (HA), neuraminidase (NA), and matrix protein 2 ectodomain (M2e), they will identify and rank candidate epitopes by determining their structural flexibility using all-atom molecular dynamic simulations, and by evaluating their conservation and surface accessibility, to promote recognition by the immune system and the generation of antibodies that target different viral strains. Candidate epitopes will be tested for antigenicity in vitro, and the best used to immunize mice to characterize the antibody response. After optimizing the vaccine regimen, they will ultimately evaluate the ability of the candidate vaccines to provide protection against diverse influenza strains in multiple animal models.

Dr. Yoshihiro Kawaoka of the University of Tokyo in Japan will develop broadly effective influenza vaccines by mixing together epitopes of conserved fragments of the viral hemagglutinin (HA) protein, which only elicit a weak immune response, together with millions of different, non-naturally occurring fragments that elicit a strong immune response, to induce broadly cross-reacting antibodies. Influenza is of world-wide concern severely impacting public health and the global economy. Tens of millions of reported cases result in tens of thousands of deaths annually in the U.S. alone, and the rapid spread of the virus between countries causes epidemic or pandemic outbreaks. Traditional vaccines are directed towards selected epitopes in the head region of the viral HA protein because they elicit a strong immune response. However, these regions are frequently mutated, rendering the vaccines useless. Vaccines directed towards more conserved epitopes only elicit a weak immune response, but this can be strengthened using HA proteins previously unseen by the immune system. They will produce a library of millions of HA epitopes that contain artificial mutations in the immune-dominant regions while preserving the conserved regions. This should focus the production of highly reactive antibodies against the conserved HA epitopes, which will eliminate a wider range of influenza strains. They will test this using single and repeat immunizations of different mixes in ferrets. Once optimized, the vaccination strategy will be tested in ferrets pre-exposed to influenza virus to mimic the human situation. The result will be a single vaccination that protects against a wider range of influenza strains than traditional vaccines.

Alice McHardy of the Helmholtz Centre for Infection Research in Germany will use a computational approach to engineer a more stable, neuraminidase (NA)-like antigen for use in influenza vaccines to increase both the duration and the breadth of protection against multiple influenza strains. Seasonal influenza viruses are constantly mutating, and current vaccines designed to target the variable, but strongly immunogenic, surface antigen, hemagglutinin (HA), must be frequently replaced. Vaccines designed to target another immunogenic antigen, neuraminidase (NA), fail to elicit a strong immune response likely due to the instability of the NA protein during the manufacturing process. They will combine genetic, structural, and evolutionary data to design an NA-like antigen with epitopes conserved across at least two influenza subtypes to provide protection against a broader range of influenza strains. The protein will be optimized for stability and immunogenicity, and then tested in mouse and ferret influenza infection models. The antigen will be co-formulated with c-di-AMP (known to improve responsiveness to vaccines in vulnerable populations including infants and the elderly) as the adjuvant. The expected result of this project is an antigen that provides increased duration and breadth of immune protection against multiple strains of influenza.

Hanseup Kim of the University of Utah in the U.S. will develop small, ultra-low power, chemical sensors that can be distributed around farms to help detect crop diseases in low-resource settings. Plants under attack from pests and diseases release low levels of volatile organic compounds that could be used as an early warning system to reduce crop losses, which can be substantial. They will design chemical sensors that trigger a change in electrical conductivity when they bind a target compound to minimize energy consumption so that they can be operated over the eight-month farming season in low-resource settings. The sensors will first be developed to bind trace levels of hexenol, hexenal, or indole, which are released from damaged maize and sorghum. They will optimize sensitivity by testing different sensor materials and correlate compound detection with different types and stages of crop damage. They will also evaluate wireless monitoring of multiple sensors distributed around a small plot of crops ready for scaling up to future in-field testing on these and other crops. Note: This grant is funded by the Foundation for Food and Agriculture Research (FFAR).

David Hughes of Pennsylvania State University, John Corbett of aWhere, and Rhiannan Price of DigitalGlobe, in the U.S. will develop a software platform comprising prediction algorithms that leverage artificial intelligence to predict where and when plant diseases and pests will occur from weather and satellite data to alert farmers to check their crops. Pests and diseases are moving targets, however most current surveillance methods monitor only their presence or absence. Predicting when and where they are likely to occur would be more valuable for preventing them. This has recently been made possible by studies on how environmental factors influence the emergence and behaviour of crop pests and diseases. They will use a systems approach that incorporates these new predictors along with historical data and couples them with an artificial intelligence component that learns from ground observations recorded using smartphones to improve accuracy. They will combine their existing agricultural intelligence platform and smartphone application with their prototype predictive model and test their approach with maize and cassava crops on farms across seven different counties in Kenya. The platform will produce location-specific forecasts that can be acted upon immediately by farmers.

There is a compelling need for the development of new drugs for trematode infections since current drugs are often ineffective and/or have widespread resistance. Drug repurposing which is advantageous in fast-tracking compounds into clinical studies is a promising drug discovery approach. Edwin’s research involves the application of computational biology in the drug repurposing of kinase inhibitors as new therapies for trematode infections

The project aims to discover molecular scaffolds that could be forerunners of EAEC therapeutics. Following a small molecule library screen, the team is evaluating hits, determining their mechanisms of action and their potential to be progressed as drug candidates. The group will also apply their anti-biofilm screen to other small libraries with a view to increasing the repertoire of promising leads against EAEC and other neglected enteric pathogens.

The project aims to develop an assay/test to measure the activity of antimalarial drugs on the transmission of the malaria parasite. This innovative work is anticipated to be a useful addition to the current tools for drug discovery and to support the malaria eradication agenda

The aim of the research is to identify selective modulators of the TB bacteria, Mycobacterium tuberculosis (Mtb), to support development of newer, more effective therapies.

Yingda Xie of Rutgers, The State University of NJ and JoAnne Flynn of the University of Pittsburgh, both in the U.S., will develop a non-invasive approach for testing candidate anti-tuberculosis compounds in animal models and patients using positron emission tomography-x-ray computed tomography (PET/CT). Tuberculosis (TB) is a leading cause of death in developing countries, and rates are sustained by the causative bacterium, Mycobacterium tuberculosis, developing resistance to current drugs. To circumvent this, new drugs are being designed to target human cells and proteins rather than those of the bacteria. To test these drugs, new tools are also needed to monitor TB in patients. 18 Fluorodeoxyglucose (FDG)-PET/CT is a non-invasive imaging tool that uses radioactively-labelled glucose to light up areas of metabolic activity in the body such as the lesions formed by M. tuberculosis and immune cells that play a critical role in infection. They have histopathological sections and cell and chemical data of TB lesions from non-human primate models and will use them to quantify the different lesions. Then, by using the available PET-CT scans of the lesions, they will search for quantitative signatures that can predict a specific type of lesion. The accuracy of these PET/CT signatures will be tested in a separate group of animals. Their study will reveal details of the TB immune response across different lesions, which could help design new treatments, and the signatures can be used to test the activity of new drug candidates in animal models and humans.

Paul de Figueiredo and Daniel Alge of Texas A&M University in the U.S. will develop a portable, disposable bioreactor for the low-cost production of gut microbial biotherapeutics at an estimated $0.09 per dose in low-resource settings. Dysfunction of the human gut microbiome is a common consequence of malnutrition in poor countries. It may be effectively treated with live biotherapeutics, yet current production methods are complicated and expensive. Glucose oxidase consumes oxygen as a co-substrate in glucose oxidation and has been shown to create hypoxic microenvironments in vitro, similar to that in the human gastrointestinal tract. They will engineer an inexpensive bioreactor by immobilizing glucose oxidase in a hydrogel placed in dialysis tubing and incubated in liquid media; the glucose oxidation reaction will deplete the bioreactor of oxygen and create an oxygen gradient to mimic the intestinal lumen. This will enable growth of a consortium of anaerobic bacteria, after which the microparticles will be removed by filtration. They will optimize the system using an artificial consortium of at least ten strains of common gut bacteria.

Jessica Manning and Fabiano Oliveira of the National Institute of Allergy and Infectious Diseases along with Daniel Parker of the University of California Irvine, all in the U.S., will develop a combinatorial approach to better define the incidence of vector-borne diseases in Cambodia in order to limit their transmission. The global incidence of vector-borne diseases such as malaria and dengue are increasing globally. However, accurately defining the disease-burden is challenging, particularly in resource-poor settings where diagnostics and expertise are limited. They will perform an exploratory clinical study in a peri-urban referral hospital with a catchment area of over 60,000 people in Kampong Speu Province, Cambodia, using a combination of approaches to define the high-resolution landscape of vector-borne diseases in the heart of the Greater Mekong Subregion. The study will involve next generation sequencing to identify pathogens in prospectively collected samples from infants with fever in their existing pediatric cohort as well as in new samples to be collected from febrile individuals aged between 0 and 45 arriving at the hospital; ELISA to generate antibody reactivity profiles of vector saliva, which plays a critical but often overlooked role in disease transmission; and geolocating cases using open-source (Quantum GIS) software. These data will be used to model transmission risk and identify target areas for interventions.

Joyce Nakatumba-Nabende of Makerere University in Uganda will use artificial intelligence to mine data from local village radio stations to generate timely data on crop pests and disease in sub-Saharan Africa. Crop loss due to pests and disease threatens the economic survival of smallholder farmers, and access to surveillance data is critically important yet often unaffordable. Local radio shows are a powerful source of information flow in rural African villages: they cover topics including politics, policy, climate, and social circumstances, in addition to crop concerns. Collectively, this information provides a holistic representation of current events in these communities. They will analyze local broadcasts to generate crop surveillance data that is linked to the local community situation. Radio content will be collected at low cost through a collaboration with Pulse Labs Kampala, and they will build artificial intelligence models based on deep neural networks and keyword identification to mine the data. The results will be combined with photographs of diseased crops provided by local farmers and used to train machine learning models to ultimately extract radio information in multiple languages and with diverse accents. This project will provide near real-time crop surveillance data and allow for timely responses to threats.

Gregory Medlock of the University of Virginia in the U.S. will develop a method to predict the optimal combinations of different strains of human gut microbes with health-promoting (probiotic) properties to maximize their yield by fermentation and minimize production costs. Microbes tend to grow better when they are in a mixed population (co-culture) because they can share resources and are more resistant to pathogens. Co-culturing can also lower production costs. However, identifying the right mix is challenging as it depends on the properties of each strain and how they interact with other strains. To simplify this, they will build an algorithm that can be applied to any strain of interest. Metabolic and growth profiles will be collected from 10 probiotic strains grown under different conditions to determine their nutrient preferences. These profiles will be used to model the metabolic space occupied by each strain for identifying the combinations that maximize the potential for cross-feeding and minimize resource competition when grown under specified culture conditions. They will test their predictions by comparing biomass produced using the predicted combinations with random combinations and with strains grown on their own.

Shabir Madhi, Vicky Lynne Baillie, and Courtney Paige Olwagen of Wits Health Consortium in South Africa will use next generation sequencing to identify pathogenic causes of neonatal deaths and stillbirths to help develop new treatments such as vaccines and better prevent disease spread. There are around 2.5 million neonatal deaths annually, and an estimated 2.6 million stillbirths, the vast majority of which occur in low- to middle-income countries. Conventional assays have recently indicated that infectious diseases cause far more of these neonatal deaths and stillbirths than previously thought. They also revealed a worryingly high proportion caused by multi-drug resistant, hospital-acquired infections. They will apply a next generation sequencing approach to 80 archived blood samples from stillbirths obtained in an earlier study and analyze the results using the Global IDseq software platform to identify any novel or unculturable or untargeted pathogens that were not identified using the more traditional microbiological methods. They will also use in silico approaches to analyze virulence factors and antimicrobial resistance profiles of identified pathogens.

Christophe Lacroix of ETH Zürich in Switzerland will develop a new method to grow mixed strains of bacteria in bioreactors more efficiently and at lower cost for producing microbial-based biotherapeutics by immobilizing the bacteria on porous polysaccharide gel beads. Damage to the naturally-occurring bacterial populations in the human gut often occurs as a result of malnutrition and can cause serious illness. Healthy populations may be restored by gut microbial biotherapeutics - the ingestion of mixtures of naturally-occurring gut bacteria. Traditional processes to manufacture these mixtures is complex and expensive because many strains have strict growth requirements and do not grow well in mixed culture. They will immobilize bacteria on microbeads to allow multiple strains of bacteria to grow in the same culture: antagonistic strains will be spatially separated, less competitive strains will be maintained, and diversity will be preserved. Using inoculums from healthy human adults, they will optimize individual components of the system including the composition of the bacterial mixture, method of immobilization, and fermentation conditions within a continuous-culture stirred-tank reactor. The result will be the controlled, reproducible and efficient production of gut microbial biotherapeutics.

Ahmad Khalil of Boston University in the U.S. will develop a low-cost bioreactor platform to simultaneously optimize growth conditions of multiple bacterial species for large scale production of biotherapeutics. The human gut microbiome plays an essential role in health and development and living microbial biotherapeutics could be an effective treatment in the case of damage by illness or malnutrition. Commercial production is limited by the capacity of bioreactors, which are costly and challenging to scale and relatively inflexible. Using their eVOLVER continuous culture system, which is modular, inexpensive, and highly scalable, they will adapt the set-up and optimize protocols to allow for the management of unique growth conditions for individual species in parallel, and dynamic mixing of cultures from individual pools to precisely tune multi-species formulations. They will conduct full-scale tests to evaluate their approach for optimizing production of human gut microbiota.

Melanie Bannister-Tyrrell of Ausvet in Australia will create an SMS-based communication system for farmers in Kenya to anonymously report crop disease and pest infestations and generate surveillance data to minimize crop loss. Pest infestation and disease cause substantial crop losses each year. In many low-income countries, farmers do not report disease to local agricultural authorities because they fear their crops will be destroyed without compensation. Yet information on the presence and spread of pests is needed to inform decisions on planting and control measures. They will optimize the design of an SMS interface for anonymous reporting using SMS codes for diseases and pests. The system will be piloted in one county, leveraging existing users and community volunteers to recruit new users. Reports will be used to estimate the local prevalence of pests and disease using modified small-area statistics. If a credible threat is detected, all users will be alerted. The system will improve connection and trust between farmers and agricultural extension workers and improve surveillance.

Mustafa Naseem of the University of Michigan in the U.S. will create an android application to present digital immunization and performance data from front-line health workers to their medical supervisors to improve vaccination coverage in Pakistan. Polio is a vaccine-preventable disease, eradicated in much of the world yet endemic in Pakistan due to poor compliance with immunization schedules. Vaccine administration in rural provinces is challenging because of understaffed, understocked, and sparsely-located healthcare centers. To improve this, the government introduced eVaccs – a smartphone-based monitoring system to track the movements and vaccinations administered by each vaccinator. However, these data remain largely inaccessible to their direct supervisors. To address this, they will develop an application to present them with relevant data in a useful format in real time. This will enable supervisors to better monitor performance, identify key challenges to comprehensive vaccination coverage, and help them better manage vaccine supplies. They will perform behavioral experiments in the field to test whether their approach positively influences vaccination coverage.

Amit Lal of Geegah LLC in the U.S. will develop battery-powered ultrasonic imagers to collect and wirelessly transmit high-resolution images of soil and airborne pests for the early detection of crop threats across large farming areas in rural Africa. Crop losses due to pest infestation negatively impact both food security and local economies. Damage caused by nematodes is particularly difficult to detect because the symptoms visible above ground are not unique and are often incorrectly attributed to deficiencies in soil nutrients or moisture. Currently the only way to test for nematodes is through root and soil samples taken after harvest, when it is too late to respond to an infestation. They will integrate complementary metal-oxide semiconductors (CMOS) with GHz ultrasonic imagers to detect nematodes and survey soil properties over large areas to detect infestations before crop damage occurs and transmit the data in real time. They will optimize the sensor technology in a controlled laboratory setting to maximize sensitivity and specificity and minimize power consumption and then transition to a farm setting for incorporating data transmission via the radio frequency wireless network.

Danya Arif, Subhash Chandir, and Qadeer Baig of Interactive Research and Development Global Limited in Singapore will develop and implement a school-based initiative in Pakistan to train adolescent girls to provide practical immunization information to parents and caregivers and thereby increase vaccination coverage in peri-urban areas. Poor compliance with routine immunization schedules puts children at risk of vaccine-preventable disease; some of the most vulnerable are urban-slum communities where traditional methods to increase vaccination uptake have failed. As an alternative approach, they propose to create a strong community force of adolescent girls, trained in leadership and communication skills and educated on basic immunization facts, to counsel mothers in their community on the importance of timely vaccination and provide them with practical information on clinic locations and schedules. The girls will also be equipped to identify never- and under-vaccinated children. They will pilot test their approach in ten schools in two peri-urban slums in Karachi city with 500 female students in grades eight to ten. They predict that mothers will be more likely to accept and act on information delivered by these girls as accepted members of their community, and training the girls as adolescents will have a sustainable impact: as they themselves become mothers they will be lifelong advocates for childhood immunization.

Subhash Chandir, Danya Arif, and Ali Habib of Interactive Research and Development Global Limited in Singapore will develop a text-based immunization chatbot that uses artificial intelligence (AI) and natural language processing to provide vaccine education and practical information to caregivers to improve vaccine coverage in Pakistan. Although free clinics exist, many children remain incompletely vaccinated because caregivers are unaware of the importance of immunization and uninformed about clinic locations and schedules. They will develop Bablibot – a chatbot to replicate interactive human conversations and able to understand caregiver queries and respond with automated text messages. The bot will provide information on immunization locations and schedules, address post-vaccination concerns, and prompt caregivers when vaccinations are due, easing the burden on overworked health workers. Chatbot conversations will be validated through human-in-the-loop (HITL) to enhance AI potential and, if needed, connect the caregiver with a human being. It will be widely accessible as it works via text. It will be pilot tested by integrating with the existing digital immunization registry of over one million children and recruiting 5,000 caregivers. The ability of Bablibot to provide accurate information, continuity of care, and convenient interaction with the healthcare system could increase immunization demand and coverage.

Paul Namwanja of the Community Health Centre, Busabala in Uganda will implement mobile technology for village health teams to monitor households and childhood vaccinations on remote islands in Uganda and to establish public-private partnerships with community health workers and boat owners to improve vaccine coverage. Vaccination rates on Ugandan islands are significantly lower than the national level because of unreliable transportation to access mainland healthcare centers; island residents rely on commercial fishing boats to travel and face long wait times for healthcare service. They will modify an existing mobile application for village health teams to create a registry of new and expectant mothers by recording household location and required vaccines. These data can then be shared with mainland health workers and used to facilitate access to a subsidized boat taxi service for travel to medical appointments. They will test their approach on six trial islands by providing the mobile phones and application along with training to health workers and recruiting boat drivers and evaluate its effect on vaccination coverage.

Rumi Chunara of New York University in the U.S. will collect data from mobile phones of healthcare workers to develop algorithms that will help prioritize healthcare resources to increase vaccine coverage in Punjab, Pakistan. Immunization is one of the most cost-effective and successful public health strategies and is estimated to prevent up to three million deaths each year. Still, many rural areas of low- and middle-income countries have an under-vaccinated population due to a lack of formal education and awareness of the importance of vaccinations. They will collect data on disease incidence and vaccine resources and coverage from the mobile phones of 3,800 rural healthcare workers. These data will be used to train an artificial intelligence model to identify the ideal times and locations to target vaccine efforts. The model will also address accessibility and awareness issues by incorporating distance to healthcare centers and frequency of visits. Their approach will be evaluated by analyzing coverage before and after implementation. Once proven, it can be scaled to other areas in Pakistan.

Rajiv Rimal of George Washington University in the U.S. will increase vaccination uptake in Nepal by improving the state of health centers to make them more enjoyable and practical places for mothers to bring their children and to motivate health workers to provide better care. In Nepal and many other low-resource settings, essential health services such as vaccinations are often provided in settings with long waiting times and limited facilities, which is also demotivating for the staff. To address this, they will interview recent mothers and service providers to understand the challenges around receiving and providing vaccinations. These will be used to design an intervention that involves improving the relationship between service providers and those giving the vaccinations as well as cleaning up the health centers by painting and providing seating. They will test the ability of their approach to increase vaccination uptake by conducting a two-arm cluster randomized study using 10 clinics in the Makwanpur district.

Do-yeon Pi of PiQuant in the Republic of Korea will develop a small, low-cost, water quality test equipment and a GIS-based monitoring system to continuously map water quality in real-time across Vietnam. Despite the extensive river network running through the country, many Vietnamese, especially in rural areas, remain short of clean water. Drinking water can be polluted by many different pathogenic microorganisms such as Salmonella and E. coli that cause waterborne diseases. Spectroscopic devices are used to monitor water quality but they require expensive time-consuming procedures to remove background noise and improve accuracy. They have built a small spectroscopic device incorporating a noise-canceling algorithm that can quickly and accurately measure water quality at a much lower cost than traditional devices. They have identified the major water pollutants in each region and have developed a prototype water scanner. This will be distributed in a pilot area to demonstrate preliminary proof of performance for measuring water quality. They will also set up a GIS-based monitoring system that maps the water quality data and allow users to analyze the potential causes of water pollution and react quickly.

Noshad Ali of Precision Health Consultants Pvt Ltd in Pakistan will develop a speech recognition platform to record child vaccination data and increase efficiency at vaccination clinics. Adherence to the recommended vaccination schedule is critical for reducing vaccine-preventable disease in developing countries and is increased when caregivers have positive interactions with healthcare workers. They will implement a system that will allow caregivers to dictate and record child vaccination information via speech recognition. This will decrease the amount of time spent recording data, allowing vaccinators more time with caregivers to address any concerns. This should help strengthen the relationship between healthcare workers and caregivers and thereby promote vaccination adherence. Software developers will shadow vaccinators in clinics during the design phase, noting the main discussion topics and questions asked. Once functioning, the success of the platform will be measured by software precision and satisfaction of vaccinators using the technology.

Bolanle Oyeledun of the Center for Integrated Health Programs in Nigeria will develop a new approach that involves teaching Nigerian parents to leverage their motivations for ensuring their children were fully immunized so that they can persuade other parents in low-income settings to do the same. They will hold interviews and group discussions with around 250 parents and grandparents of children who are fully immunized across four regions of Nigeria. These will be used to identify their positive and negative experiences during the vaccination process and why they ultimately ensured their children were fully immunized. From this group, around 40 will be selected to act as mentors who will be trained to deliver some of the key lessons learned from the interviews via group discussions. The ability of their approach to increase knowledge and demand for vaccinations will be tested with the 40 mentors along with 400 caregivers of infants younger than five months who have already missed at least one vaccination.

Federico Costa of the Federal University of Bahia, Brazil, Mitermayer Galvão dos Reis of Fiocruz, Brazil, and Nathan Grubaugh and Albert I Ko of Yale University in the U.S. will establish metagenomic next generation sequencing in clinical settings in an urban region of Brazil classified as an infectious disease ‘hot spot’ to help develop new diagnostics and identify emerging pathogens. Rapid urbanization in Salvador, a metropolitan of 2.9 million inhabitants in Northeast Brazil, has produced a favorable environment for the emergence and spread of infectious diseases, and was the founding site for the recent Zika epidemic. Early detection is critical for preventing disease spread, particularly in Salvador, which is a transport hub and popular holiday destination. However, diagnosis can be challenging in low-resource settings, especially when the causative pathogen is unknown, the disease has diverse symptoms, or a known pathogen starts causing new symptoms. They will collect around 160 clinical samples from patients with suspected infections at a local infectious disease reference hospital and a maternity unit and apply a next generation sequencing approach together with the IDseq analysis platform to identify pathogens from the sequencing data.

James Berkley and Abdi Abdirahman of the University of Oxford in the United Kingdom will test whether metagenomic analysis of clinical samples from patients with suspected infectious diseases can better identify the causative pathogens than current diagnostic methods, to help improve treatment. In Africa and Asia, many severely malnourished infants die after being discharged from a hospital likely due to infectious disease syndromes such as pneumonia, diarrhea, and sepsis. However, the causative pathogen remains unidentified in the vast majority of cases due to the limited sensitivity of current diagnostic methods. Another group highly vulnerable to the limitations of diagnostics currently available in low-resource settings are those with suspected viral encephalitis, which can cause acute seizures and coma. If diagnosed correctly, some of these diseases are treatable. They will use next generation sequencing to perform metagenomic analyses of samples from patients in both groups to identify the causative pathogens, and test whether first isolating extracellular vesicles that may concentrate viral nucleic acids from the samples can improve sensitivity.

Thushan de Silva, Abdul Karim Sesay, Helen Brotherton, and Beate Kampmann of the Medical Research Council in the United Kingdom will locally implement equipment and methods for next generation sequencing of a range of clinical sample types to detect infectious pathogens in hospitalized neonates in low-resource settings. Almost three million children under five years old die each year in sub-Saharan Africa, which is the highest rate globally. A substantial proportion is likely to be caused by pathogenic infections, including multi-drug resistant organisms. However, defining the etiology of neonatal infections, which is crucial for timely and effective treatment and to block disease spread, remains challenging in countries with limited resources. They will test the potential for next generation sequencing to overcome this challenge by conducting a pilot study using clinical samples from around 30 hospitalized neonates within an ongoing clinical trial in a low-resource setting in West Africa. They will optimize protocols on site for detecting pathogens from a range of sample types and develop methods to specifically detect relevant anti-microbial resistant strains. They plan to use the study to install a network of sites across the region that can perform next generation sequencing and share the resulting large datasets with other sites.

Ophelia Venturelli of the University of Wisconsin-Madison in the U.S. will study the growth kinetics and microbial interactions of a synthetic bacterial community in order to optimize bioreactor design and produce large quantities of mixed cultures at low cost. Mixed microbial populations are used to reconstitute the healthy gut microbiota in infants and children who have suffered malnutrition. However, affordable, large-scale production of microbial communities for biotherapeutics is challenging, in part because of poor understanding of how the growth and viability of individual strains are impacted by environmental factors and the presence of other microbes. To address this, they will build a dynamic model of a synthetic 12-member community that mimics the functional and phylogenetic diversity of the human gut microbiome and monitor its stability and growth over time and in response to variations in culturing parameters. This information will be used to predict community growth and stability in bioreactors, which will then be tested. Their approach is low cost, scalable to industrial-sized bioreactors, and can be generalized to other microbial communities relevant for human health.

Samuel Dorevitch of the University of Illinois at Chicago in the U.S. will build solar-powered ozonation systems to supply purified water to families living in Kenyan slums. Many peri-urban informal settlements (slums) around the world lack safe, affordable drinking water. In the absence of centralized water purification, methods like chlorination, solar disinfection, and filtration can be used. However, these are time-consuming and expensive, and are generally not monitored for water quality. They have developed microplasma technology for a solar-powered system that can purify surface water by ozonation without the need for electricity. They will increase the scale of this system to produce 2,000 litres of clean water each day and add sensing and reporting technology for sending information on water quality to local health officials. To test their approach, a purification system run by an array of solar panels will be installed in each of two slums in Kisumu city to provide water for 50 families and be run by local clean water teams. Two frameworks for managing and sustaining the clean water system - a membership-based cooperative and a for-profit vendor charging a nominal fee – will also be tested. The results will be used to inform the next stage of expansion to systems with a 10,000 litre/day capacity in cities in two African and two South Asian countries.