Periodic Reporting for period 1 - PathoGelTrap (New Blue Revolution through a pioneering pathogen-trapping technology based on bioselective hydrogel-forming proteins)
Okres sprawozdawczy: 2020-06-01 do 2021-05-31
Aquaculture currently supplies more than 50% of all the human consumption of fish and seafood, and is a key player to ensure future food and nutrition security, especially in poor countries where fish is the main source of proteins. To meet the challenge of feeding a rapidly growing global population, large-scale farming is needed. However, intensive farming models could drive to an increase in disease outbreaks, impacting fish health, production, the environment and the economy of this industry. Disease outbreaks could become a major constraint to the expansion of aquaculture and have a significant impact on the economic development and aquatic products supply, as well as for the environment. The number of tools available for disease control in European aquaculture is limited. The use of medication is necessarily largely controlled, and prevention programmes include a combination of the use on vaccination, water general disinfection treatments and management programmes. Sustainable solutions for preventing and controlling pathogen hazards are used in ecological aquaculture (quarantine, lower fish density, better health management, etc.). However, these solutions are sometimes difficult to transfer to intensive farming without an unaffordable reduction of industry production. In this context, new models to control and prevent pathogen infections are urgently needed for ensuring longevity and sustainability of the so-called “Blue Revolution”.
The PathogelTrap Project proposes to transform the future of aquaculture driving a change of paradigm in disease management practices. The project will provide the industry with a pioneering pathogen-trapping technology, able to target and remove specific pathogenic agents directly from the water. At the core of our science-enabled technology are 2 topics revolutionizing the scientific community: affibodies (AFBs) and the Liquid-Liquid-Phase Separation phenomenon (LLPS). These liquid-droplet forming proteins often present the so-called Low Complexity Regions (LCRs), with a lower diversity of residues, and follow a phase transition dynamic: the monomeric proteins self-assemble into liquid droplets (liquid-liquid phase separation) which later evolve into a gel state, i.e. hydrogel (liquid-to-gel transition).
The PathogelTrap Project proposes to transform the future of aquaculture driving a change of paradigm in disease management practices. The project will provide the industry with a pioneering pathogen-trapping technology, able to target and remove specific pathogenic agents directly from the water. At the core of our science-enabled technology are 2 topics revolutionizing the scientific community: affibodies (AFBs) and the Liquid-Liquid-Phase Separation phenomenon (LLPS). These liquid-droplet forming proteins often present the so-called Low Complexity Regions (LCRs), with a lower diversity of residues, and follow a phase transition dynamic: the monomeric proteins self-assemble into liquid droplets (liquid-liquid phase separation) which later evolve into a gel state, i.e. hydrogel (liquid-to-gel transition).
Inspired by this naturally occurring cellular phenomenon, we propose to develop PathoGelTrap (PGT), a dynamic tool to bind specific pathogens into the water. We use the current knowledge on the self-assembling properties of the LLPS proteins to perform rational design of a chimeric LCR-AFB biomaterial that will recognize fish pathogens (both viruses and bacteria) and trap them. We aim to provide the industry with two different solutions: i) PGT Liquid: the LCR-AFB monomeric protein as a flocculant agent to be added directly into the fish-farm water. ii) PGT Filter: we will cast a customized LCR-AFB hydrogel to be used as a preformed filter for trapping the targeted pathogenic agents when passing through aquaculture filtration systems. The project has chosen two relevant models, Betanodavirus affecting Seabass, and Yersinia affecting rainbow trout.
Viral Nervous Necrosis (VNN) also known as Viral Encephalopathy and Retinopathy (VER), is a hazardous and devastating disease of many species of cultured and marine fish worldwide. It is caused by Betanodaviruses, a serious concern especially in larvae and juvenile fish. This virus infects most of the cultured fish causing severe mortality. European sea bass (Dicentrarchus labrax L.) is a very valuable fish species in Mediterranean countries, and it is currently one of the main cultured fish species in Europe. However, different infectious diseases can affect its production and cause important economic impacts in the aquaculture industry. One of the most significant diseases affecting D. labrax is viral encephalopathy and retinopathy (VER), which is characterized by severe damage to nervous tissues.
Enteric redmouth disease (ERM) is a disease generally accruing amongst cultured salmonids, causing significant economic losses to the fish-farming industry. The disease is caused by Yersinia ruckeri, a gram-negative bacteria first isolated from rainbow trout in Idaho, USA and currently found throughout North and South America, Europe, Australia, South Africa, the Middle East and China.
Towards the use of PGT products in two selected environments, marine and fresh water, we have selected the best LCR components derived from the in vitro phase separation assays in real fish farms conditions (hCPEB3-S6, DBP1 Nt+Ct and mHP1α). Also, the selected antigens (the P-domain of Betanodavirus RGNNV and OmpF membrane protein from Yersinia ruckeri) have been defined. The new selected binder scaffold (rcSso7d) has presented good results, particularly with Betanodavirus. This will contribute to high specificity and maximum efficiency. Also, we have avoided environmental drawback via the selection of LCRs with no amyloid properties. During the screening each LCR candidate has been selected according to the expression already described in E. coli and focused on small size and high yields of production: with the subsequent scaling-up step in mind, the LCR-AFB chimera must be capable of being produced in large concentrations and at low cost.
Also the characterization of the operation conditions typical of both production environments (marine and fresh water farms) are used in the selection and design of the molecular components. This will contribute to the efficacy and the safety of the PGT products, both from the animal welfare and an environmental safety perspective.
Viral Nervous Necrosis (VNN) also known as Viral Encephalopathy and Retinopathy (VER), is a hazardous and devastating disease of many species of cultured and marine fish worldwide. It is caused by Betanodaviruses, a serious concern especially in larvae and juvenile fish. This virus infects most of the cultured fish causing severe mortality. European sea bass (Dicentrarchus labrax L.) is a very valuable fish species in Mediterranean countries, and it is currently one of the main cultured fish species in Europe. However, different infectious diseases can affect its production and cause important economic impacts in the aquaculture industry. One of the most significant diseases affecting D. labrax is viral encephalopathy and retinopathy (VER), which is characterized by severe damage to nervous tissues.
Enteric redmouth disease (ERM) is a disease generally accruing amongst cultured salmonids, causing significant economic losses to the fish-farming industry. The disease is caused by Yersinia ruckeri, a gram-negative bacteria first isolated from rainbow trout in Idaho, USA and currently found throughout North and South America, Europe, Australia, South Africa, the Middle East and China.
Towards the use of PGT products in two selected environments, marine and fresh water, we have selected the best LCR components derived from the in vitro phase separation assays in real fish farms conditions (hCPEB3-S6, DBP1 Nt+Ct and mHP1α). Also, the selected antigens (the P-domain of Betanodavirus RGNNV and OmpF membrane protein from Yersinia ruckeri) have been defined. The new selected binder scaffold (rcSso7d) has presented good results, particularly with Betanodavirus. This will contribute to high specificity and maximum efficiency. Also, we have avoided environmental drawback via the selection of LCRs with no amyloid properties. During the screening each LCR candidate has been selected according to the expression already described in E. coli and focused on small size and high yields of production: with the subsequent scaling-up step in mind, the LCR-AFB chimera must be capable of being produced in large concentrations and at low cost.
Also the characterization of the operation conditions typical of both production environments (marine and fresh water farms) are used in the selection and design of the molecular components. This will contribute to the efficacy and the safety of the PGT products, both from the animal welfare and an environmental safety perspective.
New biomolecular tools for disease control are on demand for Health Management Programmes within animal production systems. However, the success of novel, new technologies depends to a large degree on the public’s risk-benefit perception. The perceived risk from LCR-AFB biomaterial is largely based on the uncertainties related to its release and fate in the environment. With novel products, materials and processes comes potential secondary risks related to usage and disposal. The fate and behaviour of these biomaterials, maybe of ecotoxicological, and subsequently human toxicological concern. Unintended and unmanaged exposure to these biomaterials, coupled with their novel physico-chemical characteristics and behaviours, may result in adverse (eco)toxicological effects. Exposure modelling involves the collection of data relating to the characteristics, release and fate of a biomaterial and the application of this data to theory relating to environmental or biological fate and behaviour in order to quantify actual or potential environmental or human risk from the biomaterial of concern.
The use of PGT as an integral part of the biosecurity strategy of the aquaculture sector will improve the custom-made-tool-box for disease prevention. This will translate into better health management, improved animal welfare, more precise pathogen removal, reduced transfer of pathogen charge to the environment.
The use of PGT as an integral part of the biosecurity strategy of the aquaculture sector will improve the custom-made-tool-box for disease prevention. This will translate into better health management, improved animal welfare, more precise pathogen removal, reduced transfer of pathogen charge to the environment.