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PARTNERSHIPS FOR ENHANCED ENGAGEMENT IN RESEARCH (PEER)
Cycle 6 (2017 Deadline)


Mapping of pesticide residue and oocysts on vegetable and fruits using low-cost field based assays

PI: Basant Giri, chembasant@gmail.com, Kathmandu Institute of Applied Sciences
U.S. Partner: Toni Barstis, Saint Mary's College, Notre Dame, Indiana
Project Dates: December 2017 - November 2019


Project Overview:

Contamination of vegetables and fruits by pesticides and endoparasites is a major public health concern in developing countries, including Nepal. Pesticides are widely used worldwide to protect crops from pests, but their excessive and unmanaged use is harmful to humans and the ecosystem. Pesticides are well-known toxins that cause acute and delayed health effects, including disruption of the central and peripheral nervous systems and cancer. Similarly, according to the World Health Organization (WHO), about 8% of total deaths reported in Southeast Asia are caused by diarrhea. In Nepal, several instances of diarrheal outbreaks are reported every year. Human exposure to pesticides and parasites is due to contaminated food and water. Concerns over food pollution are rising in Nepal, but science-based understanding of the level of pollution is limited, due in part to the unavailability of reliable, economical, easy-to-use, and rapid field test methods to be used as important early warning tools for consumers. Conventional methods for determining pesticide residues on food involve sophisticated, time-consuming, expensive chromatographic methods that require advanced lab facilities and skilled operators. Similarly, endoparasite oocysts in food products are identified using expensive, high resolution optical microscopes, immunofluorescence-based microscopy, and polymerase chain reaction techniques that are not suitable for field screening of samples in developing countries. This project aims to develop (1) a paper-based pesticide residue assay and (2) smartphone-based oocyst assay methods. Both of these high throughput, low-cost, easy methods will be first developed and validated in the laboratory and later tested with vegetable samples at various locations across Nepal. It will be the first large-scale field testing of these methods with real samples. Other activities on the project will include providing training to students and government technicians, organizing workshops for concerned stakeholders, creating online map with field results, and disseminating information. Bringing reliable assay methods for pesticide residue and oocysts for public use with the possibility of commercialization will be the major output of the project. The USG-supported partner will provide technical advice and training for both the lab development and field testing components.

In addition, the team will involve undergraduate students from the Tri-Chandra Multiple Campus, Tribhuvan University. They will learn to use the newly developed assay methods and go into the field to test food pollution levels. In addition, two Master’s students from the Central Department of Environmental Science, Tribhuvan University, will be given the opportunity to carry out their thesis work in the framework of this project. The PI and his colleagues will share their field screening findings and involve the Plant Protection Directorate (PPD) of the Government of Nepal. PPD technicians will be trained in the new methods and be given some of the assay tools. Two workshops and one symposium will also be organized to share the results of the project.


Summary of Recent Events

In this project, the PI and his team aim to develop two types of devices/methods. The first one is a paper-based analytical device that will determine the pesticide residue in vegetables and fruits. The second one is a smartphone based microscope that will count the (oo)cysts in vegetable and fruits samples. They spent this first quarter ending March 2018 mainly in planning the project.

Planning of the project involved reviewing already published reports and journal articles related to their study. As collaboration and support from other researchers and government officials is also very important to successfully complete our work on time, they took initiative to organize and participate several meetings including the Social Welfare Council, Ministry of Science and Technology and the Ministry of Agriculture Development.

In the mean time, we finalized recruiting human resources such as research associate, project coordinator, thesis student and research interns. The team also visited a number of other research labs in Kathmandu to get an idea on the type/manufactures/model equipment these labs are using for their research work. The labs we visited were - a) Research Institute of Bioscience and Biotechnology, b) Central Department of Biotechnology Tribhuvan University, c) National College of Higher Education (NIST), d) Center for Health and Disease Studies Nepal, and e) Phutung Research Institute.

Dr. Giri also visited and had meetings with Faculty and students from Institute of Engineering, Pulchwok Campus and Kantipur Engineering College, Dhapakhel - both under Tribhuvan University regarding involving
student researchers in "smartphone application" development. To this end, students from Kantipur Engineering College have been selected for "app" development. These students are being jointly supervised by Dr. Basant Giri (PI), Dr. Bishesh Khanal from Nepal Applied Mathematics and Informatics Institute (NAMII) and Dr. Suresh Manandhar from the York University, UK. These students have made some progress on their work.

Lastly, they have also started initial testing of pesticide detection assays. The pesticide residue detection method is based on enzyme inhibition activity of pesticide. The enzyme they proposed to use is acetylcholinesterase. To this end, they tested two different types of enzyme substrates (indoxyl acetate and Dithionitrobenzoic Acid). They are also optimizing various parameters of the enzyme assays such as the enzyme concentration, signal reading time, solvent effect etc. In all these testing, they used Dichlorvos (DDV) as standard pesticide. The optimization will require more work.

In the next 3-6 months, the activities will involve getting final letter of approval from SWC, complete planning and purchasing equipment, purchasing regents and supplies. During this period, they hope to get a first demo of smartphone app, and complete optimization of enzyme assay and start lab work on (oo)cyst detection.

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