Team:IOANNINA/Implementation

Implementation

Proposed Implementation

Our team designed the basic parts of a microorganism that can deactivate tetracyclines and macrolides in its environment. Our proposed implementation revolves around poultry waste management. This microorganism will deactivate the bioactive antibiotics that are present, before they end up in the environment, which could further cause AMR.

We chose to focus on poultry waste for two reasons. Firstly, our city, Ioannina, is located in Epirus, an area where chicken and animal farms are thriving. These farms are the biggest in our country, supplying more than half of the chicken products that are consumed. Secondly, poultry waste usually undergo a specific management procedure, with fertilizer as the final product.

During this procedure the bioctive antibiotics are in direct contact with the soil and they can add pressure to the bacteria present towards developing antibiotic resistance mechanisms. This leads to an increase in antibiotic resistance genes (ARGs), thus amplifying the problem of AMR.

Our goal is to minimize the amount of antibiotics before they end up in the environment. More specifically, we minimize the time antibiotics remain bioactive in the environment and at the same time we create an antibiotic free raw material, ready for further processing.

After considering all the safety standards and consulting experts in the field, we came up with a possible implementation system for our engineered bacterial strain. This bacterium (not our chassis organism) will not be harmful for humans and animals, and it could belong to the flora of the poultry, so no disruption of the micro-communities during the poultry waste management will occur.

A suitable suggestion is the Gram-negative bacterium called Labrys portucalensis F11. This bacterium has been isolated from antibiotic-contaminated soils and from sediments, sludge, animal feces and seawater. , (Cycoń et al., 2019). Concerning the biosafety level, it belongs to Risk group 1 (Schoch et al., 2020).

Nowadays, the process that is followed in Greece regarding poultry manure management includes 3 stages. For the first stage the poultry manure is removed from the hennery and transported to a specific open place (i.e., to the ground), where is let to become homogenous for 2-4 months in piles. This is the stage where the manure gets into contact with the soil.

During the second stage, composting occurs via the activation of the existing microorganisms and the rise of temperature. This stage lasts approximately one month. This is the time required for the raw material to reach 70°C for at least an hour.

This stage takes place in several long channels, at the start of which the poultry manure is loaded in. In the end of these channels, the poultry manure has its final composition and, after desiccation (stage 3), it can be sold as fertilizer in the form of pellet.

So, after having discussed this matter with an expert (Mr. Tzamixas, manager of Fertilizer Factory and Rendering Plant), we suggested that a bioreactor be inserted in the existing line of the waste management. More specifically, as soon as the poultry manure is removed from the farm, it will be inserted in a bioreactor containing our engineered microorganism. The bioreactor will stir the whole mass, letting the microorganism come in contact with as much antibiotic as possible. When all the antibiotics are deactivated and the microorganism is dead, the bioreactor will be emptied and the next batch will be loaded into it and so on. After this treatment, the stages of the poultry management will be followed as described above. By doing this, we will minimize the time that the antibiotic containing manure is in direct contact with the soil, causing the emergence of ARGs in the bacteria present in it. In other words, we aim to solve the problem at its roots.

Surely, there are many things to be done before the actual implementation of this project. We are only in the first stages of this proposed solution for the problem of AMR. There are many aspects that should be taken into consideration, such as how big the bioreactor should be, how much time the engineered microorganism needs to deactivate all the antibiotics, how we may ensure biocontainment. However, we feel we have proposed a solid design that others may build upon.