Meagre and the greater amberjack are promising emerging species in aquaculture. This is due to their fast growth rate, large size, high processing yield and high quality fillet. However, little is known about the immune system of these fish and several potential pathogenic threats to sustainable culture of these fish have been identified. Therefore, this thesis has begun to characterise the immune response of these species in order to better combat pathogenic threats. The focus of this thesis is the identification, modulation and monitoring of key innate and adaptive immune genes. This is achieved by identifying conserved regions of target gene sequence and designing consensus primers to these in order to generate a partial sequence, which could then be used as a base from which RACE PCR could be performed to obtain full sequence. The identified sequence was then used to produce qPCR primers, anti-meagre IgM and anti-meagre IgT monoclonal antibodies and a functional synthetic greater amberjack piscidin active peptide. Using the developed qPCR primers the constitutive expression of target genes was determined in the gills, gut, head kidney and spleen. Changes in transcript expression were then monitored in these tissues in response to PAMP stimulation in vivo and in vitro in order to better understand how these genes are modulated by viral, bacterial and fungal stimuli. Furthermore, the meagre development of the immune system was monitored from 1 day post hatch (dph) until 120 dph, providing insights into when immune maturity is achieved. Monoclonal antibodies were also produced and their viability tested in a variety of assays including Western blot, Immunohistochemistry and ELISA. Finally, A synthetic greater amberjack piscidin was produced and the bacteriostatic capabilities of this peptide were tested and showed promising results against known bacterial fish pathogens, indicating a potential for future uses within and outwith aquaculture.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:752666 |
| Date | January 2018 |
| Creators | Milne, Douglas John |
| Contributors | Secombes, Chris ; Zou, Jun |
| Publisher | University of Aberdeen |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://digitool.abdn.ac.uk:80/webclient/DeliveryManager?pid=237613 |
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