FishPhytoFeed

Title: Development of an Advanced Integrated Toolbox for in vitro high-throughput screening of quality and phyto-estrogens in feed ingredients for Mediterranean finfish

Funding Source: EPAnEK Aquaculture

Budget IMBBC: 70,000€

Start / End Date: 2020 - 2023

Web site URL: https://fishphytofeed.gr


Project Progress: 100%

Principal Investigator:

Efthimia (Efi) Cotou

Project Members:

Research Directions:

Fish nutrition and feeding

Biomarkers and bioassays

Description:

The use of alternative ingredients in fish feeds calls for the development of adequate methodology for the control of their quality.Current methodologies are of analytical character and lacking the capacity to actually determine the dimension of their biological effects. The ultimate goal of the FishPhytoFeed is the development of an advanced integrated toolkit for the in vitro high throughput screening of fish feeds ingredients based on their biological activities and effects. No use of live fish will be required and the toolkit determinations will be combined in a synthetic quality factor that will reflect the effects of ingredients on key biological processes that consist fish growth.

Project Overview

The existing methodologies for controlling the quality of fish feed are based on feed analysis and resort to the determination of concentrations of substances that are possibly involved in the biological effects of plant raw materials, but without ultimately being able to determine the true dimension of these biological effects. The present project will use classical enzymology methodologies to determine the presence of KTI and BBI protease inhibitors, the pH-stat method to determine the digestibility of dietary proteins in vitro by sea bass and sea bream digestive enzymes, the Microtox Assay System© to determine of generalized cytotoxicity of raw materials, the ER-CALUX assay in human bone marrow cell lines with an integrated luciferase gene and the YES assay© in yeast cells bearing estrogen receptors for the determination of estrogenic activity of plant raw materials, and primary muscle cell cultures of sea bream to determine their myostatic action.

Aim

The ultimate goal of the program is the development of a comprehensive raw material testing toolbox that will allow rapid and efficient evaluation of a large number of raw materials for their biological activity without requiring experimentation with fish. The individual metrics of the toolkit will be combined into a composite feedstock quality index that will reflect feedstock impact on key biological functions that constitute fish growth.

Expected Results

The successful implementation of the project will lead to:

  • - Introduction of certified methodologies for the advanced quality control of raw materials and fish feed that will not require tests on fish and will produce quick conclusions regarding the biological action of the raw materials
  • - exploitation of knowledge on the effect of the most widespread raw materials used in the breeding of Mediterranean species on cytotoxicity and on the processes of digestion and growth of white muscle
  • - Identification of marker genes that signal muscle growth in sea bream and indicate its growth course
  • - Recording of estrogenic activity and concentration of genistin in the most widespread raw materials used in the breeding of Mediterranean species.