Theses and Dissertations


Title: Identification of Channel Catfish (Ictalurus Punctatus) and Basa (Pangasius Bocourti) Fillets by an Enzyme Linked Immunoabsorbant Assay using Monoclonal Antibodies and by Fatty Acid Methyl Ester Analysis

Name: McNulty, Shawn Thomas

Degree: PhD

Chair: Klesius, P.

Resides: SFAA

University: Auburn University

Location: Auburn

Date: 2004

Pages: 0

Keywords: channel catfish, basa, pangasius, flesh identification, monoclonal antibodies

Abstract:

The first objective of the study was to produce
monoclonal antibodies (MAb) that would identify both channel catfish and basa
fillets. A monoclonal antibody specific for a 36.8 kDa channel catfish (Ictalurus
punctatus 
) fillet protein
was produced and characterized by indirect enzyme-linked immunoabsorbant assay
(ELISA) and western blotting. Using the MAb clone 1A10 in the indirect ELISA,
fillet proteins from Ictalurus
sp. 
could be differentiated
from non-Ictalurid fish fillet proteins. One hundred percent of Ictalurus fillets were correctly identified in a
single blind study of both raw and cooked fillets. An additional MAb that was
specific for a 71.6 kDa basa fillet protein was also developed. The MAb, clone
3F11, bound specifically to fillet proteins of the genus Pangasius in an indirect ELISA and correctly
identified basa fillets in a single blind study. These results show that
indirect ELISA using MAbs is a rapid and sensitive method for identification of
channel catfish and basa fillets. The second objective of this study was to
determine whether fatty acid analysis using software developed by MIDI (Newark,
DE) was able to identify channel catfish fillets and differentiate from basa
fillets. Ten samples of 25 mg frozen muscle from each species were processed to
produce fatty acid methyl esters (FAME) and the fillet fatty acid profiles were
used to create species-specific fillet libraries. The results show that the
fatty acid profiles of basa are significantly different from those of channel catfish.
The fatty acid fillet profiles for each species were highly reproducible when
the same fillet was analyzed daily for three consecutive days. In a single
blind study, 25 fillet samples from each species were assayed to identify the
species of the fillet sample processed. The FAME method identified 25 of 25
(100%) basa fillet samples. Three out of 25 samples from channel catfish
fillets were incorrectly identified as basa fillets. However, none of the basa
fillets were identified as channel catfish fillets.

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