Work Packages

This project is split between five work packages which aim to cover all aspects of evaluating prophylactic health products and novel alternatives in small-scale aquaculture farming. These include value chain analysis, product efficiency and microbial safety, as well as ensuring that the project has a lasting impact on stakeholders.

WP1: Project management and dissemination

The University of Stirling is responsible for the management of the project including operational, technical and financial aspects. Alongside partner countries it will also manage country relations and ethical monitoring, monitoring gender, social and ethical issues encountered during the project implementation as well as those raised by research.

Work Package 2: Scoping and overview of production systems, PHP markets and usage

T2.1 Systems overview of farming systems and sample-frame development

Collect, collate and appraise available data on smallholder intensification, associated farmer livelihoods and aquatic production systems for target commodity-species and countries.
Case-studies on key species/ country systems using rapid rural appraisal techniques.

T2.2 Survey of farmer health management practices and PHP usage

Review of secondary data and development of farmer surveys to assess current PHP use patterns as part of wider farm health-management challenges and practices
This survey will also incorporate a qualitative component designed to assess under-lying motives for farmer decision behavior with respect to application of PHPs and associated trends e.g. opinion of product efficacy, use history, cost-benefit perceptions compared to alternatives etc.

T2.3 Value Chain analysis for PHP products including regulatory assessment

Collect, collate and appraise available data on PHP markets, governance, policy, regulatory systems, labelling and certification schemes
Application of Global Value Chain (GVC) framework for assessment of market dynamics, power relations, lead companies and their quality specifications, processing, storage and distribution characteristics, seasonality factors, margins, risks and rewards of participation
Value chain analysis will be based on key-informant and stakeholder semi-structured interviews.
Results will be used to identify ‘up-grading’ options at appropriate levels of value-chains with a view to directly or indirectly benefiting target beneficiaries

T2.4 Inventory of commercial PHPs

Development of a photographic and written inventory of prphylactic health products (PHPs) on the market
Information on composition, provenance, supply, price, storage and distribution to be collected
Information will be compared to data collected in Tasks 2.2 and 2.3.
Results will be used to classify commercial products according to a range of product attributes e.g. derived from farmer opinion, market availability, cost and the robustness of product claims.

T2.5 Microbiological and chemical analysis of PHPs

Samples of PHPs belonging to different product classes identified in T2.4 will be evaluated in the laboratory for mode of action
Information on composition, provenance, supply, price, storage and distribution to be collected
Results will be used to shortlist a range of the most promising products as candidates for use as positive controls in RCT trials (T3.2)

WP 3: Evaluating cost and benefits of commercial probiotic PHPs

T3.1 Assessment of the effect of PHPs on immune functionality

To evaluate the mode of action of PHPs in fish/ shrimp, we will use larval zebrafish to model changes in gut histology, microbiota and immune parameters
An assessment of impact of PHP upon gut health, microbiota and fish immune response will be produced

T3.2 Randomised Control Trials (RCTs)

RCT trials will be conducted with smallholder farmers to compare the efficacy of selected probiotics (T2.5)
In aerated shrimp ponds probiotic use will also be compared against untreated ponds with C:N optimised heterotrophic biofloc assemblages.

T3.3 Evaluation of PHPs on organoleptic qualities of tilapia in Bangladesh

Samples of tilapia will be collected from RCT trial ponds (T3.2) in Mymensingh District for assessment of organoleptic qualities through taste testing.
Different parts of the tilapia carcass consistent with local eating habits will be selected for analysis. This will include samples most likely to contain the highest concentrations of off-flavour causing geosmin and MTB due to their elevated fat content.
Samples will be steamed and their flavour assessed by a trained panel of 10 trained panel of members using standard processing industry evaluation scales for different taints.

T3.4 Evaluation of the prevalence of foodborne pathogens in ponds and the microbiological safety of probiotic strains

The prevalence and antibiotic sensitivity of foodborne pathogens in case vs control ponds will be assessed, in order to anticipate potential issues of transfer of antibiotic resistance genes.
Any probiotic strains identified as carrying antibiotic resistance genes will be ruled out of further analysis and PHP development
The probiotic strains will also be tested in cell culture systems or in experimental animals, as appropriate, for their toxicity towards fish and humans

WP4: Evaluating novel pre-biotic carbohydrate decoy and natural product immune stimulatory PHPs

Marine organisms have the potential for transforming into Prophylactic Health Products in aquaculture practice. In addition, several Indian medicinal plants are already effectively administered as PHPs in shrimp/fish aquaculture. Typical bioactives are generally either extracts or compounds of bacterial, fungal, microalgae, macroalgae or animal origin. They are mainly polysaccharides, peptides and peptidoglycan, alkaloids, saponins, phenolics, terpenoids etc.

T4.1 Sourcing, extraction and isolation of carbodydrate-based bioactives

Selected mangroves, marine microalgae, seaweeds, sponges and echinoderms will be sampled from marine environments along the Indian coast
These will include marine species such as Microalgae (Chlorella spp., Phormidium valderianum), Mangroves (Bruguiera spp, Xylocarpus spp., Cereops tagal), Algae (Ulva, Gracilaria spp., Hypneamusciformis, Laurencia spp., Portieria hornemannii, Porphyra., Sargassum spp., Turbinaria spp.,), and selected echinoderms. Selected terrestrial plants with established medicinal value and immunomodulating effect will also be sampled (e.g. Withania somnifera, Ocimum sanctum, Artocarpus heterophyllus, Bidenspilosa, Urticadioica, Warburgia ugandensis, Fagaropsis angolensis, Sapindus laurifolius).

T4.2: Development of carbohydrate and small molecule microarrays

Glycan and natural product arrays will be developed with materials provided by Asian partners and visitors.
These microarrays will be based on the substances listed above

T4.3 Assessment of microbial interactions with microarrays

With validated methods in hand, evaluation of fish pathogen binding to arrays will conducted in the lab with microbes from the zebrafish gut microbiome models established in WP3 or directly with WSSV.
These analyses will probe the arrays with cultures of individual species of fish/shellfish pathogen (e.g. Streptococcus agalactiae, S. iniae and Flavobacterium columnare, Edwardsiella ictaluri, E. tarda, Aeromonas hydophila, and WSSV), and mixed microbiome cultures, to assess which organisms recognise which carbohydrates.
This will identify which bioactives bind to pathogens.

T4.4 Elucidation of bioactive structures

We will drill down to analyse which glycans and natural products are present in the array 'hits'
This will define structures of carbohydrate/natural product-pathogen interactors.

T4.5 Evaluation of the efficacy of identified bioactives under lab and farm conditions

‘Hits’ from microarray analyses and prospective immune-active methanol extracts will be evaluated for their efficacy and mode of action in vivo in a zebrafish model

T4.6 Development of standardised laboratory operating procedures (SOPs) for producing and evaluating the efficacy of PHPs

Standardisation and scale-up of pertinent glycan and natural product extractions will be established, along with recommended species/cultivars and formulations. SOPs will be worked-up for roll-out to farmers, feed manufacturers and suppliers, and/or directly to fish farmers or, depending on the outcomes above and the technical complexity or otherwise of the process.

WP5: Technology transfer and commercialization

The project will use a modified theory of change approach to supporting pathways to impact and a participatory market chain approach (PMCA) to disseminating technologies and improving market access for the resource-poor. The PMC approach aims to foster market access by generating collaboration among the different market chain actors and to facilitate change in market chains that lack coordination. The approach aims to create an environment that builds trust among the different market chain actors through supporting mutual learning and shared innovation.

This will be delivered through:

Establishment of an online network for knowledge exchange and dissemination of outputs to farmers, extension services, SMEs, pharmaceutical companies, certification and regulatory bodies etc. The network will be also used to establish case-study dialogues between probiotic users and other value chain members with different perspectives over the project life. It will also be used a vehicle to engage with networks of related sectors (eg livestock health and nutrition)
Exchange visit of key actors in PHP value chains in Bangladesh and India
SME workshops in partner countries on novel PHPs
Dissemination of key messages at National Farmers meetings
Development of prophylactic health management extension materials for farmers
Policy briefs on regulatory needs and recommendations
Data archiving
Media dissemination of key findings