BioMatNet Item: FAIR-CT96-1518 - Development of microalgal pigments for aquaculture

FAIR-CT96-1518
Development of microalgal pigments for aquaculture

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FAIR Area 1.2 - Green Chemicals and Polymers Chain : Fine Chemicals

	Proposal No:	FAIR-CT96-1518
	Date Prepared:	July 2001, November 1999, September 1999
	Source:	Final Report Abstract and Executive Summary Progress Report September 1999 Progress Report September 1998

Final Report Executive Summary

Final Report Abstract

Source: Final report, February 2001

Consortium: The project was co-ordinated by the School of Biological and Earth Sciences, Liverpool John Moores University (UK), Universidade Catolica Portuguesa, Escola Superior de Biotechnologia, Porto (Portugal), Universidad de Santiago de Compostella, (Spain), Necton Campanhia Portuguesa de Culturas Marinhas Lda, Olhao (Portugal) and the Instituto de Ciencias Biomedicas Abel Salazar, Porto (Portugal), PEP Research & Consultancy Ltd, Plymouth University, Devon (UK), Addavita Ltd., Chesterfield (UK), Sorgal, Sociedade de Oleos e Racoes SA, Porto (Portugal), Bionova, Santiago (Spain).

Abstract

Objectives

The global objectives of this project are:

to improve the efficacy of feed formulations for the use of algal astaxanthin in the Aquaculture industry, and
to scale-up methodologies for the production of fresh-water microalgal biomass/pigments.

Activities

The work programme for this research and development project focused on the following technical activities. The first objectives were the selection of a suitable species and strain of microalgae that could be used for the commercial production of astaxanthin. Next, optimal conditions for the growth of the organism and identification of the conditions required for astaxanthin production were to be identified and conditions optimised. One of the most important tasks was the development, construction and subsequent testing of closed photobioreactors that would be suitable for the mass cultivation of the alga (and permit product accumulation). Identification of the equipment and conditions for the downstream processing of the astaxanthin-rich alga was also a very important objective as this directly affects product quality and subsequent efficacy. Finally the use of the product(s) developed would be tested in both salmonids (e.g. Rainbow Trout) and selected Seabream species. These pigmentation trials would be performed against existing commercial products and would ensure that commercial levels of pigmentation could be achieved in the target species.

Achievements

The main achievements of this project are:

Selection of suitable species and strain of microalgae for astaxanthin production
Optimisation of algal performance in terms of improved productivity and yield
Development of a novel media to support optimal microalgal yields
Identification and optimisation of conditions for astaxanthin accumulation
Development of alternative closed photobioreactor designs (suitable for scale-up) for the mass cultivation of microalgae (for use indoors and outdoors)
Optimisation of conditions for harvesting of algal biomass (green and red)
Optimisation of conditions for the processing (cracking / extraction / drying) and storage of the astaxanthin-rich algal biomass
Demonstration of the efficacy of algal astaxanthin products in pigmentation of salmonids (rainbow trout)
Markets for natural astaxanthin and new commercial opportunities identified

Together these have enabled an industrial process for the production of natural astaxanthin to be developed.

Progress Report September 1999

Objectives

The overall objectives of this project are:

to improve the efficacy of feed formulations in the use of algal astaxanthin in the aquaculture industry, and
to scale-up methodologies for the production of fresh-water microalgal biomass/pigments.

Activities

A series of tasks and sub-tasks have been established in order to achieve the main objectives of this project. Following on from progress in year one, in the second year of the project, the research and development work concentrated on the following activities.

The continuing development and testing of novel photobioreactors for outdoor cultivation in a closed system at an appropriate operational scale.
Identification of the ideal strains of astaxanthin-accumulating microalgae
Determination of optimum conditions for batch, fed-batch and continuous cultivation of microalgae (partly identified in year one) in larger scale cultures.
Development of a new (industrial) medium for mass cultivation of the identified strain for biomass/astaxanthin production

In parallel with the work directly related to the algae, the efficacy of natural (algal) astaxanthin was assessed in both rainbow trout and in gilthead seabream. An in vitro bioavailability system was established, with factors affecting the bioavailability of algal astaxanthin identified and actions taken to improve bioavailability and biodeposition of astaxanthin tested.

Progress

This has been in line with expectation. Following the original design of two prototype novel closed photobioreactors in year one, the design and operation of these systems was further developed and a number of operational problems overcome. The reliability and robustness of these bioreactors has been greatly improved, although further work will be needed in year three of the project. The optimal conditions for microalgal growth and carotenogenesis in batch and fed-batch culture for use in these bioreactors have been identified. Feed trials of both Gilthead Seabream and Rainbow Trout with astaxanthin-rich Haematococcus have been performed. These studies indicated that microalgae rich in astaxanthin can readily contribute the desirable (marketable) levels of pigmentation in Rainbow Trout, and thereby provide a natural alternative to the current synthetic product. An in vitro bioavailability system using isolated fish guts has been developed.

Achievements

The main achievements during the second year of this project are:

identification of an optimal microalgal strain for the synthesis of astaxanthin
development of a new industrial growth media to optimise algal biomass/astaxanthin production
further development of two novel closed photobioreactors for optimised microalgal growth
extensive testing of the reactors
the incorporation of algal astaxanthin into extruded diets has been shown to result in marketable levels of pigmentation in Rainbow Trout
a market analysis study for astaxanthin utilisation has been undertaken.

Future Activities

These will focus on three main areas.

The development of the phobioreactors for outdoor algal cultivation under controlled conditions, with more trial work using Haematococcus, as well as further work to improve the processing of Haematococcus

In addition, the processed materials produced need to be evaluated as to their efficacy in terms of fish pigmentation. Hence, further feed trails have been planned with both rainbow trout and seabream using processed astaxanthin-rich microalgae and purified carotenoids in order to understand the limits of bioavailability and thereby further improve bioavailability of the carotenoid. In parallel with these feed trials, a series of in vitro studies using isolated inverted fish guts will also be undertaken to understand bioavailibility of algal astaxanthin. The effects of carotenoid supplementation in the diet of rainbow trout on fish health will be assessed, since this activity was postponed from year two.

Progress Report September 1998

Summary

Objectives The global objectives of this project are:

to improve the efficacy of feed formulations for the use of algal astaxanthin in the aquaculture industry
to scale-up methodologies for the production of fresh-water microalgal biomass/pigments

Activities Seven tasks and associated sub-tasks have been established in order to meet the main objectives of this project. In the first year of the project, the bulk of the research and development work was focused on mass outdoor cultivation of freshwater microalgae. To achieve high yields and to minimise or eliminate contamination and predation, novel closed photobioreactors were to be designed and constructed. The optimum conditions for batch, fed-batch and continuous cultivation of microalgae within these systems were to be identified. The conditions optimal to the use of microalgae rich in astaxanthin as a pigment source for marine and freshwater species of commercially farmed fish were also to be determined.

Results Progress towards the overall objectives within the first 12 months of the project has generally been on-target and met expectations. Facilities for the cultivation of freshwater microalgal species that accumulate astaxanthin have been constructed. This included the design, building and characterisation of two prototype novel closed photobioreactors which will be used to produce carotenoid-rich microalgal biomass. These bioreactors have been developed on the basis of trial runs with and without microalgae. Preliminary data indicate that these systems are capable of producing the required high levels of biomass and are efficient for carotenogenesis. The optimal conditions for microalgal growth and carotenogenesis in batch and fed-batch culture for use in these bioreactors have been identified. Feed trials of both gilthead seabream (marine) and rainbow trout (freshwater) with astaxanthin-rich microalgae have been performed. These studies indicate that microalgae rich in astaxanthin can be an effective source of pigmentation provided that the algal cell is processed to achieve desirable levels of bio-availability.

Progress The main achievements during the first 12 months of this project are:

the optimal conditions for batch and fed-batch cultivation of Haematococcus to produce astaxanthin have been identified
feeding trials using carotenoid-rich microalgae as a pigment source have been performed for rainbow trout (freshwater) and gilthead seabream (marine)
a series of prototypes for novel closed photobioreactors for microalgal growth have been designed and constructed and are being tested
a market analysis study for astaxanthin utilisation has been undertaken.

Future activities The technical work to date has highlighted the need for considerable attention to the processing of astaxanthin-rich algal cells in order to improve its bio-availability. As a result this task has been given higher priority. The next 12 months will also see the first complete scale-up trials in the UK and Portugal of the prototype closed photobioreactor algal growth systems with a working volume of 400+ litres. In addition, further feed trails will be conducted with both rainbow trout and gilthead seabream using processed microalgae and purified carotenoid esters. The effects of carotenoid supplementation in the diet of rainbow trout in terms of the state of fish health will be determined. A series of in vitro studies using isolated fish organs will also be undertaken. The objective here will be to further improve the deposition efficacy of microalgal carotenoids within the flesh and/or skin, thereby improve the overall performance of microalgal pigment products.