Vector-borne diseases are a major global health burden. One of the most dangerous <B>vectors</B> is the <B>mosquito</B>, which is responsible for more than <B>one million</B> deaths annually. After taking a blood meal on an infected individual, the mosquito carrying the <B>pathogenic agent</B> can transmit the pathogen to another host during a new bite. Despite intense research, <B>vaccines</B> and/or <B>treatments</B> are still needed, thus, the fight against these deadly diseases relies mostly on <B>vector control</B>. One of the primary sources for wide spread vector control are the usage of <B>insecticides</B>. </br>However, the over spraying of insecticides impacts the <B>environment</B> and leads to the selection of insecticide <B>resistance<B> mosquitoes. Therefore, we developed a novel diagnostic device, <a href="https://2016.igem.org/Team:Pasteur_Paris/Moskit_devices"><B>Mos(kit)o</B></a>. This kit includes a <B>fixed or mobile mosquito trap</B> and a <B>biosilica cellulose composite patch</B>, incorporating proteins produced by bacteria, which was the result of using genetically, modified <i>E. coli</i>. It was our intent to create a device that not only <B>classified</B> the vector-borne viruses circulating within specific regions but also provided a tool for local agencies to <B>identify hot spots</B>. This would allow for better <B>targeting</B> of insecticide spraying and it could help address public and environmental health concerns.</br></br></br>

With the assistance of <B>synthetic biology</B>, we successfully modified <i>E. coli</i>, within a <B>contained</B> fully functioning biosafety laboratory. With it, we produce the protein needed to create biosilica, and bind antibodies onto a cellulose support. We selected <B>biosilica<B> to increase <B>rigidity</B> of our patch and because it is completely <B>biodegradable</B>. The innovative design of the patch creates a multilayered matrix coated with antibodies capable of <B>detecting</B> a wide panel of vector-borne pathogens and insecticide resistant proteins from captured mosquitoes. This patch is <B>customizable</B> and can be easily adapted to simultaneously test for multiple vector-borne pathogens prevalent in specific locations. Additionally, the patch will have <B>2D barcoded</B> readouts, generating an environmental <B>surveillance database</B>. A precise map of vector hot spots will provide a better assessment and response to vector-borne diseases, assisting <B>local health authorities</B> with <B>anticipating</B> and <B>preparing</B> for an epidemic. </br></br>

The development of our project’s concept and the design of our device (i.e. patch and trap) was an iteractive process that involved a multidimensional collaboration of people and ideas. We successfully achieved this through:</br>

4) consulting local agencies (French Interdepartmental Mosquito Control Board (EID) and in the United States the New Orleans Mosquito Control Board) and a drone company to discuss our project and if applicable, potential collaboration.</br></br>  

Additionally, we incorporated surveys assessing the general public’s understanding and acceptance of synthetic biology. These steps were essential to ensuring we gathered knowledge, gained feedback, and considered as many options as possible; making necessary adjustments based on our findings. Our tool will be user-friendly, safe, and applicable.</br> </br></br>   

The Mos(kit)o device is easy to use and it does not require scientific expertise so it is possible to train people locally. From the set up of the trap to the analysis of the patch, the steps are simple and designed to reduce user error. For example, the quick read function has a color change component to ensure quick and accurate interpretation of results; changing from no color to color for positive results.</br> </br>   

  The students working on this project received laboratory safety training and were supervised during all experiments. All manipulations involving GMOs only took place in a contained biosafety level II laboratory and the usage of GMOs was only required for the production of the biosilica protein (made using E. coli bacteria). The protein was made in the cell and when the cell was lysed, the protein was released and the GMO destroyed. Therefore, there was no release of the GMO during this process. </br></br>

In the Field. All personnel using the Mos(kit)o will be trained to ensure proper usage of the device. The kits are designed to be safe and user friendly, requiring no scientific background. The operators will not have any contact with the infectious materials, reagents, or the newly produced protein. </br></br></br>

Upon successful completion of our lab work and following informed discussions with stakeholders, we illustrated two application scenarios where the Mos(kit)o would be utilized: in developed countries and in developing countries. The diagram below explains the life cycle of our device, beginning with the production of the biosilica protein and concluding with real life applications and potential outcomes.</br></br>

<B>Context 1</B> describes developed countries where agencies or municipalities like the Centers for Disease Control and Prevention (CDC) in the US or the French Interdepartmental Mosquito Control Board (EID), respond to emergencies. A warning is received from a specific location and these agencies are responsible for distributing the Mos(kit)o and ensuring that appropriate personnel are deployed to endemic areas. </br></br>

Currently, the CDC has an Epidemic Intelligence Service (EIS) that responds during public health outbreaks. They are responsible for identifying the causal agent(s) and they are tasked with assisting endemic areas with efforts to prevent and/or control the transmission of the disease(s). 2 A similar institutional set-up can be found in France. Following our conversation with a division director for mosquito preparedness at EID (acronym), we developed a practical example illustrating the utility of the Mos(kit)o device. Presently, administrators manually test endemic areas, which is labor and time intensive. The design of our device eliminates these conditions by using stationary traps and a patch capable of simultaneously detecting multiple viruses. Additionally, the division director of EID expressed interested in our device and suggested the need to have these traps within certain distances from hospitals where infected people are treated or airports because it is a transit place for global entry/exit from endemic areas. Our device would be used as an important proactive measure that could protect non-infected people if they are in close proximity to these particular hot spots (e.g. hospitals, airports) where infected mosquitoes are circulating. In scenario 1, these organizations have the resources to respond to the hazards, administer the kits, and analyze the results; making them ideal for Mos(kit)o.</br></br>

<B>In Context 2</B> , which describes developing countries where access is not as easy and there are no local agencies as mentioned in scenario 1, the Ministry of Health, or equivalent instance, would assume responsibility for administering the kits. Personnel at the local Ministry of Health will be trained to properly set up the trap and on how to use the diagnostic device. We are aware that resources are limited in these settings; therefore, we designed the device to require minimum personnel for operation and to be low cost, user friendly, and safe. We were unable to discuss this specific scenario with a health ministry official to get their feedback, however, based on a conversation with tropical entomologists and the New Orleans Mosquito Control Board, which is solicited by the Brazilian government, we envisioned a situation in which a country receives a report of an outbreak of a vector-borne disease, like zika virus. In that case, the Ministry of Health would respond by requesting a Mos(kit)o device, either from the Mos(kit)o company or possibly a local NGO or CDC, with a stationary trap (for easily accessible areas) or a drone (capable of reaching remote areas). Upon arrival of the kit, trained local authorities will implement the trap in targeted locations and perform the required tests. An additional authorized person reads the results and the data will be entered into mapping software, capable of producing real-time updates.  </br></br>

In both scenarios, the trained personnel will only have to inject specific syringes containing the reagents needed to test for various viruses, having no direct interaction with the mosquitoes, which are contained within the trapping device. Following use, the device will be returned to the distributing municipality/agency where it will be washed and reloaded. All waste will be properly treated and disposed of to eliminate the release of any pathogens in the environment. The data will be uploaded into a computer database for further analysis.

The data generated from the Mos(kit)o diagnostic device could be used as a global tool assisting with the environmental surveillance of vector-borne diseases. For example, the World Health Organization (WHO) is equipped with the resources to best manage, store, and protect the data. The WHO is responsible for supporting, responding, and communicating important information related to health to globally protect people. Our novel diagnostic device will:</br> 1) assist with identifying regional hot spots allowing local authorities to quickly respond;</br> 2) provide pertinent information pertaining to specific areas to target during insecticide spraying; </br> 3) prevent the propagation of infected mosquitoes. </br></br> </br></br> </br></br>