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Edy Satria
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Establishment and maintenance of a colony of the Old World Screwworm fly Chrysomya bezziana at Balitvet in Bogor, West Java, Indonesia ., Sukarsih; Partoutomo, Sujitno; Tozer, Robert; Satria, Edy; Wijeffels, Gene; Riding, George
Indonesian Journal of Animal and Veterinary Sciences Vol 5, No 3 (2000)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (410.062 KB) | DOI: 10.14334/jitv.v5i3.193

Abstract

A colony of the Old World Screwworm fly, Chrysomya bezziana, has been established and maintained at Balai Penelitian Veteriner, Bogor, West Java, Indonesia since 1994. Culture on a convenient, defined diet able to sustain long term reproducibility of insect performance is described. The long-term viability of the colony has been further assured with an effective and reliable culturing procedure, with the genetic diversity and robustness of the colony being maintained through regular infusions of wild-type field isolates. The colony has been a reliable source of larval material used for the isolation and identification of protective antigens in the ACIAR funded Screwworm Fly Vaccination Project. Larvae have been used in the development and routine application of assays employed to evaluate vaccine efficacy. The colony continues to be a valuable resource for both screwworm fly vaccine and other new research includi the evaluation of screwworm fly attractants and lures.   Key words: Chrysomya bezziana, culture, colony, larvae, vaccination
Fractionation, identification and vaccination efficacy of native antigens from the screwworm fly, Chrysomya bezziana Riding, George; Muharsini, Sri; Pearson, Roger; ., Sukarsih; Satria, Edy; Wijffels, Gene; Willadseni, Petter
Indonesian Journal of Animal and Veterinary Sciences Vol 5, No 3 (2000)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (246.005 KB) | DOI: 10.14334/jitv.v5i3.194

Abstract

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  roteins as recombinants in a form suitable for vaccination studies.   Key words: Chrysomya bezziana, peritrophic membrane, vaccination, amino acid sequence, serine protease
Fractionation, identification and vaccination efficacy of native antigens from the screwworm fly, Chrysomya bezziana George Riding; Sri Muharsini; Roger Pearson; Sukarsih .; Edy Satria; Gene Wijffels; Petter Willadseni
Jurnal Ilmu Ternak dan Veteriner Vol 5, No 3 (2000): SEPTEMBER 2000
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (246.005 KB) | DOI: 10.14334/jitv.v5i3.194

Abstract

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  roteins as recombinants in a form suitable for vaccination studies.   Key words: Chrysomya bezziana, peritrophic membrane, vaccination, amino acid sequence, serine protease
Establishment and maintenance of a colony of the Old World Screwworm fly Chrysomya bezziana at Balitvet in Bogor, West Java, Indonesia Sukarsih .; Sujitno Partoutomo; Robert Tozer; Edy Satria; Gene Wijeffels; George Riding
Jurnal Ilmu Ternak dan Veteriner Vol 5, No 3 (2000): SEPTEMBER 2000
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (410.062 KB) | DOI: 10.14334/jitv.v5i3.193

Abstract

A colony of the Old World Screwworm fly, Chrysomya bezziana, has been established and maintained at Balai Penelitian Veteriner, Bogor, West Java, Indonesia since 1994. Culture on a convenient, defined diet able to sustain long term reproducibility of insect performance is described. The long-term viability of the colony has been further assured with an effective and reliable culturing procedure, with the genetic diversity and robustness of the colony being maintained through regular infusions of wild-type field isolates. The colony has been a reliable source of larval material used for the isolation and identification of protective antigens in the ACIAR funded Screwworm Fly Vaccination Project. Larvae have been used in the development and routine application of assays employed to evaluate vaccine efficacy. The colony continues to be a valuable resource for both screwworm fly vaccine and other new research includi the evaluation of screwworm fly attractants and lures.   Key words: Chrysomya bezziana, culture, colony, larvae, vaccination
Co-Authors Gene Wijeffels Gene Wijffels George Riding Petter Willadseni Robert Tozer Roger Pearson Sri Muharsini SRI MUHARSINI Sujitno Partoutomo Sukarsih .