Graduate Thesis Or Dissertation
 

Fish oral antigen delivery system development and optimization

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  • Oral administration is the most practical way of vaccination of fish. It has advantages over intraperitoneal injection and immersion methods that include no stressful handling, no interference with routine fish husbandry and antigens can be delivered to the fish in their feed. An oral antigen delivery system was developed for salmonides using fluid-bed spray dry technique. The delivery system was prepared by spraying antigen on to nonpareil sugar beads to form an antigen layer, then an enteric protected polymerpolymethacrylic acid-ethylacrylate (Eudragit®-L30D) layer was applied. The physicomechnical characteristics of the delivery system and antigen release was evaluated by scanning electronic microscope and in vitro dissolution testing. The delivery system was optimized in terms of enteric protection, antigen release rate and the storage stability. It was observed that the film processing temperature should be above 40°C to have a good quality enteric film coat, and the film thickness should be 29μm to ensure the enteric protection. Different types and amounts of plasticizer were tested to differentiate their effects on the antigen release. The efficacy of immunization of oral ECAMs was carried out by comparing four different administration routes in Naive juvenile Coho salmon (Oncorhynchus kisutch): oral ECAMs, intraperitoneal injection, immersion and anal intubation. Fish given a high oral dose of trinitrophenylated lipopolysaccharide (TNP-LPS) had significantly higher antibody levels than the immersion method and negative control could induce at 4 and 6 weeks post immunization. Also the serum antibody titers of fish elicited by oral enteric-coated trinitrophenylated keyhole limpet (TNP-KLH) was significantly greater than that of negative control but in a dose dependent manner. Oral immunization of a variety of antigen forms may have the equivalent efficacy as other conventional vaccination methods. Modification of oral ECAM by incorporation of sodium glycolate starch increased antigen release rate when the delivery system was at a high pH environment. The dual functionality of sodium starch glycolate, i.e., outstanding water wicking capacity and good swelling properties facilitate the antigen release from ECAMs and may improve their in vivo performance in fish.
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