Team:Vilnius-Lithuania/Implementation

According to the Food and Agriculture Organization of the United Nations, 35% of global aquaculture harvest is either lost or wasted1. Considering that fish are purposefully killed for only two main reasons - consumption and disease control - pathogens are responsible for a large portion of the economic losses2. If an accurate treatment method would be applied as soon as the first infected fish is observed, there would be no need to kill potential carriers of the disease. The ability to detect the pathogens at the very start of an infection could help drastically minimize these numbers. Our pathogen detection kit was created for exactly this reason.

To help coldwater fish farmers alleviate the burden, we propose them to be the end users of our detection system. The test is especially relevant to them, because most of the farms do not have access to expensive laboratory equipment, yet the pathogens need to be detected as fast as possible, in order to prevent further spread of the disease. The requirement for the testing systems to be user-friendly and inexpensive for the target customers are stated in the WHO guidelines3, to which we referred to and complied with while developing the system.

The vision of user interaction starts with a typical consumer of fish. As one reads about the health benefits of fish products and is influenced by the growing popularity of seafood, one naturally invests more into them. His/her friends and family are also influenced by the same factors and, together with the growing population of the world, they create a considerable demand. The aquaculture farmer adjusts their production rates accordingly. An increasing number of consumable fish suddenly starts to drop. There are noticeable changes in the behavior of some fish as well as physical differences. From the yellow accumulations around the gills it is clear that Flavobacterium has infected the farm. A farmer orders a FlavoFlow testing kit, takes a sample from the gills of the lethargic animals and within an hour or two knows exactly which species of flavobacterium is causing the fluctuations in the production rate. Now, the most accurate treatment method can be chosen and applied.

In addition to this, we created a software tool OnFlow to speed up the process of pathogen detection test development, as there are numerous pathogens in the industry that lack a quick testing system. Hence, another group of end users would be people working in the industry of pathogen detection tests. Our vision is that they could use our software for that purpose. Developers would put the information gathered from various articles into the system and it would then calculate specific parameters, such as test line distance, test reaction time and optimal volume of the sample, adjusted to the pathogen of interest. They could use the given information for the design of the test strip, saving a lot of time and resources necessary for the development.

To implement our project into the real world and make it easy to use in the aquaculture farm, we developed a 3D printed case for the testing strip. The model can be printed out by anyone, who has access to a 3D printer, making the process even more effortless. In addition, a lyophilized mixture and a universal HDA methodology for the development of our product was used, which is easy to adapt and implement for different testing goals 4. IsoAmp is also known to have the smallest amount of pipetting steps, thus reducing human error. By combining the software, the case and universal methodologies we designed an overall user-friendly and adaptable testing system.

Although the test is highly accurate, one of the primary issues that would need to be considered is the temperature at which the reaction takes place. The problem is that farmers might not have a device to keep the temperature at a constant level. This aspect should be taken into account when developing other tests and presumably considering providing a hardware device for temperature control for the farmers.

Concerning the safety aspects of this project, the bacteria that cause fish disease and can be detected with our test are not pathogenic to humans. Furthermore the samples for the test are taken from the gills of the fish, without harming the animal waiving the need to kill it.

Briefly, we developed a rapid testing kit, that provides a quick and convenient pathogen detection solution for fish farmers and a software tool that enables other developers to speed the process of testing kit development.

References

  1. FAO. The State of World Fisheries and Aquaculture 2020. 2 (FAO, 2020)
  2. Hastein, T., Scarfe, A. & Lund, V. Evaluación por métodos científicos del bienestar de los animales acuáticos. Revue Scientifique et Technique de l'OIE 24, 529-547 (2005)
  3. Lorenzo-Ferreiro Carmen., et al. “Nucleic-Acid Lateral Flow Assay Optimization with Different Gold Nanoparticle Size for Detection of Pathogen after PCR, Using L. Monocytogenes as Model. Acta Scientific Microbiology 1.10 17-24 (2018)
  4. Biolabs, N. How to design the primer and select the amplicon for IsoAmp II Universal tHDA Kit? | NEB. International.neb.com at ' '