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INDONESIA
Indonesian Journal of Animal and Veterinary Sciences
ISSN : 08537380     EISSN : 2252696X     DOI : -
Core Subject : Health,
JITV (Indonesian Journal of Animal and Veterinary Science),  ISSN: 0853-7380 E-ISSN: 2252-696X is a peer-reviewed, scientific journal published by Indonesian Center for Animal Research and Development (ICARD). The aim of this journal is to publish high-quality articles dedicated to all aspects of the latest outstanding developments in the field of animal and veterinary science. It was first published in 1995. The journal has been registered in the CrossRef system with Digital Object Identifier (DOI) prefix 10.14334.
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Articles 7 Documents
Search results for , issue " Vol 5, No 3 (2000): SEPTEMBER 2000" : 7 Documents clear
cDNA library construction and isolation of genes for candidate vaccine antigens from Chrysomya bezziana (the Old World Screwworm fly) Voucolo, Tony; Supriyanti, Florentina; Muharsini, Sri; Wijffe, Gene
Jurnal Ilmu Ternak dan Veteriner Vol 5, No 3 (2000): SEPTEMBER 2000
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (309.028 KB) | DOI: 10.14334/jitv.v5i3.195

Abstract

The construction and use of cDNA libraries for the isolation of genes encoding candidate antigens for use in a recombinant vaccine against Chrysomya bezziana is described. RNA was isolated and mRNA purified from first and third instar larvae of Chrysomya bezziana and used in the synthesis of two cDNA libraries in the bacteriophage vector λ ZAP express®. These libraries were screened using Digoxigenin-labeled DNA probes obtained from two independent approaches. First, a homolog approach used probes designed from previously characterized peritrophic membrane genes identified from the related myiasis fly, Lucilia cuprina. Secondly, a de novo approach used amino-terminal and internal peptide sequence information derived from purified Chrysomya bezziana peritrophic membrane proteins to generate DNA probes. Three peritrophic membrane genes were identified and characterized. Chrysomya bezziana peritrophin-48 was identified using the homolog approach and, Chrysomya bezziana peritrophin-15 and Chrysomya bezziana peritrophin-42 were identified using the de novo approach. The identification of these genes as encoding candidate antigens against Chrysomya bezziana has allowed the production of recombinant proteins for use in vaccination trials.   Key words: cDNA library, peritrophin, peritrophic membrane, Chrysomya bezziana, Lucilia cuprina, vaccine
Localization of the glycoprotein Cb42 in larvae of the screwworm fly Chrysomya bezziana (Diptera: Calliphoridae) Eisemanni, Creig; Muharsini, Sri
Jurnal Ilmu Ternak dan Veteriner Vol 5, No 3 (2000): SEPTEMBER 2000
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (520.363 KB) | DOI: 10.14334/jitv.v5i3.200

Abstract

The glycoprotein Cb-peritrophin-42 was localized in third instar larvae of Chrysomya bezziana using primary antibodies raised against a non-glycosylated bacterial recombinant form of this protein. Both immunofluorescent antibody techniques on unfixed whole mounts of gut tissues and immunogold electron microscopy techniques on ultra-thin sections of fixed and embedded tissues were employed. The protein was shown to be exposed over the whole of both surfaces of the peritrophic membrane and to occur throughout its thickness. Immunogold labelling indicated that Cb-peritrophin-42 was expressed in the peritrophic membrane-secreting cells of the cardia, a specialized peritrophic membrane-forming organ situated at the junction of the foregut and midgut. The accessibility of Cb-peritrophin-42 present in intact peritrophic membrane to the primary antibodies used in the immunofluorescent antibody localization indicates that this glycoprotein is a potential molecular target for vaccination of host animals against larvae of Chrysomya bezziana.   Key words: Cb-peritrophin-42, Cb42, glycoprotein, peritrophic membrane, immunolocalization
Bacterial expression of larval peritrophins of Chryosomya bezziana Wijffels, Gene; Voucoloco, Tony; Muharsini, Sri; Supriyanti, Florentina
Jurnal Ilmu Ternak dan Veteriner Vol 5, No 3 (2000): SEPTEMBER 2000
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (411.091 KB) | DOI: 10.14334/jitv.v5i3.196

Abstract

Three candidate antigens, Chrysomya bezziana peritrophin-48, Chrysomya bezziana peritrophin-15 and Chrysomya bezziana peritrophin-42, were prepared for recombinant protein production in Escherichia coli using a variety of expression vectors. Cb peritrophin-48 was expressed as a recombinant protein possessing a carboxy-terminal hexaHis tag. Cb peritrophin-15 was expressed as both a glutathione S-transferase fusion protein and as an amino-terminal hexaHis tagged protein. The glutathione Stransferase Cb peritrophin-15 construct produced a heterogeneous group of fusion proteins. Cb peritrophin-42 was also expressed as an amino-terminal hexaHis tagged protein. The two putative domains of Cb peritrophin-42 were also separately expressed, again with amino-terminal hexaHis tags. Cultures of the hexaHis constructs Cb peritrophin-48, -15 and –42 were demonstrated to be useful for the production and purification of these protein antigens and were scaled-up for vaccine trials and protein characterization studies.   Key words: Chrysomya, peritrophic membrane, peritrophin, recombinant protein, hexaHis tag, GST
Vaccination against Chrysomya bezziana: a summary of current knowledge Willadseni, Peter
Jurnal Ilmu Ternak dan Veteriner Vol 5, No 3 (2000): SEPTEMBER 2000
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (125.226 KB) | DOI: 10.14334/jitv.v5i3.201

Abstract

Methods for the culture of the Old World Screwworm Fly Chrysomya bezziana on a small laboratory scale, the production of life stages and tissues suitable for vaccine study, and the in vitro and in vivo assessment of vaccine effects have all been developed. These methods would be applicable to further studies on screwworm control by many alternative technologies. It has been shown to be possible to induce dramatic immunological effects on feeding larvae and to isolate and characterise individual antigens. However, a recombinant vaccine remains elusive. Current understanding of the effects of vaccination is discussed and suggestions made for possible future research directions.   Key words: Chrysomya bezziana, Old World screwworm fly, vaccine, recombinant antigen, sheep
An investigation of the feasibility of vaccinating against the Old World Screwworm fly Chrysomya bezziana Willadseni, Peter; Partoutomo, Sujitno
Jurnal Ilmu Ternak dan Veteriner Vol 5, No 3 (2000): SEPTEMBER 2000
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (127.591 KB) | DOI: 10.14334/jitv.v5i3.192

Abstract

The problem of the Old World Screwworm fly Chrysomya bezziana and the limitations of current methods of control are discussed briefly. Any attempt to investigate the feasibility of vaccinating against the fly, as a novel control technology, demands the establishment of methods for fly culture, the production of sources of antigens and the development of assay techniques suitable for the assessment of vaccination effects. This must be coupled to a strategy for vaccine development. This strategy is described, as a prelude to a series of papers evaluating the feasibility of vaccination in detail.   Key words: Chrysomya bezziana, screwworm fly, vaccine
Expression in yeast (Pichia pastoris) of recombinant Cb-peritrophin-42 and Cbperitrophin- 48 isolated from Chrysomya bezziana (the Old World Screwworm fly) Muharsini, Sri; Voucoloco, Tony
Jurnal Ilmu Ternak dan Veteriner Vol 5, No 3 (2000): SEPTEMBER 2000
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (343.863 KB) | DOI: 10.14334/jitv.v5i3.197

Abstract

Pichia pastoris has been investigated as a means to express recombinant forms of two putative peritrophic membrane antigens from Chrysomya bezziana, Cb-peritrophin-42 (Cb42) and Cb-peritrophin-48 (Cb48). Recombinant Cb48 was expressed as a secreted and glycosylated protein. The yield of recombinant protein was 8 mg per litre of culture. In contrast, recombinant Cb42 was not expressed at detectable levels in Pichia pastoris, probably due to A + T rich sequence which may cause premature transcriptional termination. To expedite Cb42 expression in yeast, Cb42 was divided into two domains: Cb42A and Cb42B. Cb42B was successfully expressed in Pichia pastoris, yielding 0.4 mg per litre of culture. However, Cb42A was not expressed. This work demonstrates that although Pichia pastoris offers considerable benefits as an expression system producing high level of glycosylated protein, success may vary from protein to protein.   Key words: Chrysomya bezziana, peritrophin, Pichia pastoris
Vaccination trials in sheep against Chrysomya bezziana larvae using the recombinant peritrophin antigens Cb15, Cb42 and Cb48 ., Sukarsih; Partoutomo, Sujitno; Wijffels, Gene; Voucoloco, Tony; Willadsen, Peter
Jurnal Ilmu Ternak dan Veteriner Vol 5, No 3 (2000): SEPTEMBER 2000
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (149.842 KB) | DOI: 10.14334/jitv.v5i3.199

Abstract

Recombinant forms of a number of peritrophic membrane proteins from the screwworm fly Chrysomya bezziana have been assessed in vitro and in vivo for their efficacy as antigens in vaccination against the tissue-invasive, larval form of the parasite. The proteins included Cb15 and Cb42 expressed in Escherichia coli and Cb48 expressed in both Escherichia coli and Pichia pastoris. In all cases, the in vitro assays of larval growth on serum from vaccinated sheep failed to show inhibition of larval weight gain or any detrimental effect on larval survival relative to controls. Chrysomya bezziana Cb48 has a significant degree of sequence identity with the antigen PM48 from Lucilia cuprina. Feeding Lucilia cuprina larvae on antisera to Cb48 induced a small but statistically significant reduction in weight gain, as does feeding on antisera to PM48. In vivo, larvae feeding on sheep vaccinated with Escherichia coli-expressed Cb15 and Cb42 and Pichia pastoris-expressed Cb48 showed marginally greater weight gain and survival which was equal to or greater than that on non-vaccinated sheep. The significance of these observations is discussed.   Key words: Chrysomya bezziana, recombinant antigen, peritrophin, vaccination

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