User Manual

Lastly, we designed the instruction manual that includes all needed information in a simple yet precise manner for anyone to use. We aimed to streamline the process by naming all solutions used as S1,S2 and S3 and we also included a key for the interpretation of results. It is our belief that we simplified the process as best as we could without sacrificing any sort of scientific validity.

Charting

As with every virus, TSWV has its own distinct epidemiology, affecting some countries more than others. That is the reason that we sought to pinpoint the countries most heavily affected by the pathogen. After a thorough search we stumbled across some very well upkept databases that included data on the spread of tospoviruses around the world as well as their future projected status. A great model country to study the epidemiology of TSWV would be Egypt since it occupies a large portion of the worldwide tomato production but data, compared to other countries, are not easily retrievable.

According to FAO and Hagar Saeed, journalist of the “Egyptian Gazette”, the area of agricultural land in Egypt is confined to the Nile Valley and delta, with a few oases and some arable land in Sinai. The total cultivated area estimations range from 7.2 to 11.5 million feddans (1 feddan = 0.42 ha), depending on the year and the definition of what constitutes a cultivated land. Agriculture is a major component of the Egyptian economy, contributing 11.3 % of the country’s gross domestic product. The agricultural sector accounts for 28 % of all jobs and over 55 % of employment in Upper Egypt is agriculture-related.

It is important to note that the extent of the damage that solely TSWV has caused is unknown, yet it surely contributes to the plummet observed.

Nonetheless, one should always keep in mind that TSWV also affects pepper, lettuce, potato, papaya, groundnut, tobacco and chrysanthemum crops but most importantly, TSWV has an extremely wide host range with more than 1300 wild and weed species^5,6. This includes plant species in 15 monocotyledonous and 69 dicotyledonous families and one family of the Pteridophyta making it a major danger in our efforts for environmental conservation. Given that a plant can be a TSWV host even post-harvest makes it extremely easy to spread as the following map indicates

It is our personal opinion that even some countries not highlighted are affected by the virus yet either the presence of the virus has gone undetected or the spread has happened recently and data is still not available.

In any case there is no doubt that this virus is a global concern that, for now, affects some countries more.

Distribution

With an ideal candidate country in mind (Egypt) we sought out to identify the best possible way of distribution to it and what would that hypothetical deployment would require.

Distribution methods are heavily limited by the fact that the test includes live cells. Even though the cells, per se, are fixated and thus minimal danger is posed in regards to their survivability, the fact that they remain alive is enough of a liability for transportation standards. This is why we sought to learn how live material is preserved and transported in countries like Egypt and China. After short communications with people on the transportation sector we were informed that each country has its own distinct laws when it comes to importing and/or quarantining live material from other countries. We decided that it would be more realistic to focus our efforts in our neighbouring country of Egypt since actual transportation of our manufactured test there would be more realistic.

After reaching out to transportation companies and legal advisors we came to the conclusion that it would be wiser to ship our tests in bulk. Each test kit would be put in a hermetically sealed envelope and all envelopes would be placed in a 20*20*20 centimetre box. Each box would fit 15 kits and the average cost of transportation after enquiring about the cost in 8 different transportation companies was calculated to be 65.45 € thus amounting to a transportation cost of 4.36 € per kit. In addition to the aforementioned cost of production of the kit the grand total for the production and delivering of the test is 5.46 € .

Up next we decided to investigate the intricacies involved with transporting genetically modified microorganisms (GMMOs or GMMs). Depending on the mode of transportation, the country of origin and arrival, laws may vary. In our case we had to cover each part separately.

Departing from Greece

The laws of Greece regarding the export of GMMOs have been replaced and are in total alignment with the ones proposed by EU. More specifically according to “Directive 2001/18”, “Regulation 1829/2003”, “Regulation 1830/2003” and “Directive 90/219/EEC” even though the deployment of such organisms in EU soil is strictly regulated and a final decision on its usage may take up to 10 years. However, regarding transportation laws regulate that a legal entity is to be always held accountable for the transportation after a thorough quality check and an approval system that is yet to be deployed in its final form. After consulting with various experts on the field we have been informed that such an approval process may take up to a few years, giving us enough time to perfect our project and minimise any risk.

Transportation

Rules regarding the transportation of GMMOs are set by the IATA Dangerous Goods Regulations (62nd Edition) that is effective since the 1st of January 2021. Genetically modified organisms and microorganisms which do not meet the definition of toxic or infectious substances (such as ours is) must be assigned to code UN 3245. Boxes and individual contents must be shipped following Packing Instruction P904 (ICAO/IATA PI959) with a proper shipping name. This can be UN 3245 followed either by “GENETICALLY MODIFIED MICRO- ORGANISMS” or "GENETICALLY MODIFIED ORGANISMS".

Arriving in Egypt

According to the Library of Congress of the U.S “Egypt has no restrictions on releasing genetically modified organisms into the environment. In March 2008, the Ministry of Agriculture approved the domestic cultivation of genetically modified corn, and the Egyptian Ministry of Agriculture allowed the importation of twenty-eight tons of genetically modified corn seeds into Egyptian markets. However, in the spring of 2009, genetically modified corn seed imports were halted so that the National Biosafety Committee (NBC) could complete the country’s National Biosafety Framework (though the NBC continued to permit the planting of locally produced biotech seeds in newly reclaimed areas)”. Moreover, via our own research we could not find any special laws that have been drafted and voted by local or national assemblies on the importation and deployment of GMMOs.

In conclusion it is our firm belief that realistically speaking, the most austere measures that need to be abided to are the ones set by the E.U, yet the lack of appropriate laws by the Egyptian government must not take away from the serious nature with which we need to design our test kit in order to avoid any accidental deployment of our living yet fixated genetically modified e-coli cells in Egyptian soil.

Once all requirements are met it would be wise to look to more countries on their respective laws. However, it is a good practice to always abide to the strictest of laws regarding GMMOs and build the detection kit accordingly in order to avoid future obs.

Final Goal

Years 7-10

As we have already stated, our final goal is to provide a viable solution for conservation efforts around the globe. After successfully completing the aforementioned experiments our team would find itself in an advantageous position; by having established our tool in the field of agrodiagnostics our team would have the necessary funds to redirect towards conservation efforts. By using the income generated from crop conservation efforts we can:

1.   Conduct the experiments needed for the repurposing of our kit to diagnose even more phytopathogens

2.   Establish permanent task force teams in places of interest for reconnaissance expeditions in places of rich wilderness

1. Repurposing

Even though TSWV, and tospoviruses in general, certainly pose a great threat to biodiversity, a plethora of phytopathogens exist that have the potential to harm wild plants or they already have. Microorganisms such as Phytopthora infestans, Ips typographus, Anoplophora chinensis, Thaumetopoea pityocampa and many more are becoming an increasing threat in forests, especially the ones in the Holarktis region. According to the European Commission’s (DG ENV) final report on the disturbances of EU forests caused by biotic agents (2012)3 these pathogens exhibit an aggressive spread and demonstrate a large spectrum of minima and maxima of survivability. Accordingly in Neotropis and Paleotropis, several dangerous pathogenic species have been identified. Notably in the Amazon Forest, certain soils reveal susceptibility to phytopathogens and lower fungal community dissimilarity than other forests4 thus making it a prime candidate for the proliferation of harmful microbial species.

Repurposing the detection kit won’t be an easy task. Nonetheless, given that our R&D team will only have this single task as their only preoccupation, combined with their projected gained expertise throughout the past 7 years, a proper full-out repurposing seems realistic.

2. Taskforces

Without a doubt, phytopathogens infiltrating and destroying the wildlife flora are a global problem. That fact, combined with the open trade of people, materials and plants creates the need to address this problem in a global scale.

Of course one team of 15 people cannot do that by themselves. It is thus proposed that task force centres are established in every continent, with permanent personnel, fully funded by the income of our agrodiagnostics section of the project and with the approval of every respective country.

These centres will be in charge of receiving, distributing and conducting the diagnostics test, while gathering data on each pathogens epidemiology within the country but in neighbouring states as well. Along with governments, they will work together to draft new conservation policies and promote the importance of environmental conservation in their local societies.

After meetings with members of organisations that include kindred taskforces in their organisational plan, we reached the conclusion that each centre would require at least the following:

• A General Manager
• A scientific team of at least 10 people in charge of testing
• A team in charge of logistics and transportation
• A team of at least 2 data analysts
• A legal team of at least 3 people
• A team in charge of communication and public media engagement
• An obligatory diverse team acting as the liaisons of the taskforce and the general public

Choosing the countries where those taskforces would be set up was not an easy feat as many parameters had to be put under consideration including but not limited to the severity of the state of the local wildlife, the legal prerequisites for realising such a task, the costs of setting up a team there and the ease of access from and to neighbouring countries. For each primary taskforce choice presented in the map below a short explanation is provided:

Elaboration on our primary taskforce locations

Brazil

Brazil was one of the most abundantly clear candidates for setting up a taskforce due to its extremely rich biodiversity, the importance of its tropical forests worldwide, its location in the continent of South America, its large population and its large contribution in the worldwide plant produce economy.

Italy

Italy presents itself as a prime candidate for Mediterranean and Alpine biodiversity with rapid geological switches between ecosystems. Due to its prime location in the centre of Southern Europe, its proximity to Greece and the fact that it is a member of the European Union, Italy is an ideal place for setting up our first outsourced European taskforce.

DR Congo

The Democratic Republic of Congo is one of the most rapidly developing countries of the African continent. It was partly chosen for the same reasons Brazil was, that is, for its prime geographical position and its vast rainforests.

India

India presents some unique features that little to no countries possess in the globe. Its unique biodiversity, unusual crop produces, extremely rich sociocultural background and the growing public interest on environmental conservation strategies make it a place in the project’s portfolio that can widen possible target plants while being a testing ground for locating the very much needed balance between agrodiagnostics and conservation diagnostics.

Australia

Australia is a vast country that entails many unique ecosystems ranging from desserts to rainforests. Due to its unusual fauna it comprises a major source of information on the constant correlations and interactions between flora and fauna and how our efforts on plant conservation may aid animal conservation as well. Moreover, Australian laws are extremely favourable in matters that revolve around the preservation of their ecosystems.