The simple immunoprecipitation method was used to isolate tilapia immunoglobulin (Ig) for immunization in order to produce monoclonal antibodies (MAbs) specific to tilapia Ig. First, the tilapia antiserum against bovine serum albumin (BSA) was prepared by peritoneal injection of BSA into tilapia, and the tilapia anti-BSA antiserum was used to precipitate BSA to form the Ig/BSA immune complex. The Ig/ BSA immune complex was then injected into Swiss mice for hybridoma production. After fusion, three hybridoma clones producing MAbs specific to the tilapia antibody were selected by dot blot and Western blot. All MAbs (101A, 59G, and 11A) were bound specifically to the heavy chain of immunoglobulin M (IgM). The MAbs 101A and 59G demonstrated twofold higher affinity than MAb 11A and the commercialized antibody. However, MAbs 11A could also bind to the heavy chain of IgM in Asian seabass, Lates calcarifer, as well. These MAbs can be used to monitor the immune responses of individual fish by indirect ELISA upon exposure to various antigens.
Chitosan and chitooligosaccharides were extracted from white-leg shrimp shells by chemical treatment. Low molecular weight (13 kDa) and a high degree of deacetylation (54.83%) in chitooligosaccharides led to high water solubility compared to chitosan. Antimicrobial assays indicated that chitosan and chitooligosaccharides exhibited marked inhibitory activity against food-borne pathogenics, spoilage bacterial, and fungal strains tested. However, chitooligosaccharides revealed greater inhibitory effects than chitosan on tested microorganisms. The substitution of flour by chitosan or chitooligosaccharides in bread formulation (1 g/100 g total weight basis) showed antimicrobial effects against and sp. growth. Also, the fruity odor in bread containing chitosan or chitooligosaccharides was delayed. Interestingly, the bread containing chitooligosaccharides showed a stronger inhibitory effect against and sp. compared to bread containing chitosan and control, where and sp. were observed growing on the surface of bread after 4 days of incubation at 30 °C.
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