03163     2200241   4500001002100000005001500021008003900036035002000075082001500095084001500110245033500125250003800460260000900498300003200507520206100539650009602600700014002696700001602836700001302852700001902865700002202884990001502906INLIS00000000001769520211004095001211004                |          | |    a0010-0721002227  aARTVET2041  aARTVET2041  aFractionation, identification and vaccination efficacy of native antigens from the screwworm fly, Chrysomya bezziana. /cRiding, George; Muharsini, Sri[Research Institute for Veterinary Science Jalan R.E. Martadinata 30, P.O. Box 151, Bogor 16114, Indonesia]Pearson, Roger; Sukarsih; Satria, Edy; Wijffels, Gene; Willadsen, Peter.  aJurnal Ilmu Ternak dan Veteriner.  c2000  aVol.5 (3): p.150-159. 2000.  aProteases of larval excretory/secretory material and proteins of the peritrophic membrane are the most intensively studied
sources of potential protective antigens from the sheep blowfly
Lucilia cuprina
. Their importance in the screwworm fly
Chrysomya bezziana has now been investigated. Purified serine proteases from Chrysomya bezziana were tested for their
potential as vaccine antigens in sheep, efficacy being assessed by
in vitro
 and
in vivo
 assays with larval
 Chrysomya bezziana
. No
effect of vaccination was observed by the
in vitro
 assay. However, in the
in vivo
 challenge, larval weights were diminished in the
vaccinated sheep, although larval recoveries increased marginally. Vaccination with Chrysomya bezziana peritrophic membrane
does induce an effective immune response against the parasite resulting in a significant reduction in larval growth and
considerable larval mortality in the in vitro assay. Sequential fractionation of the peritrophic membrane with various surfactants
and chaotrophic agents of increasing solubilisation capacity resulted in the separation of discrete groups of proteins. The groups
of fractionated proteins were tested in a vaccination trial in sheep with vaccine efficacy assessed by in vitro assays. The urea
extract, guanidine-HCl extract and SDS soluble fraction each induced significant levels of protection against Chrysomya
bezziana larvae but the effects were poorer than those obtained from vaccination with whole, native peritrophic membrane.
Several major proteins selected from the three most protective fractions were purified by SDS polyacrylamide gel
electrophoresis. Since insufficient quantities of these proteins were available for vaccination trials, they were either sequenced
directly from the N-terminus or subjected to endoproteinase Lys-C digestion, followed by peptide purification and amino acid sequencing. This gave the information necessary for the expression of several of these proteins as recombinants in a form
suitable for vaccination studies. 4aChrysomya bezziana, peritrophic membrane, vaccination, amino acid sequence, serine protease  aMuharsini, Sri[Research Institute for Veterinary Science Jalan R.E. Martadinata 30, P.O. Box 151, Bogor 16114, Indonesia]Pearson, Roger  aSatria, Edy  aSukarsih  aWijffels, Gene  aWilladsen, Peter.  aARTVET2041