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Jurnal Ilmu Ternak Veteriner

JITV
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Published by Indonesian Center for Animal Research and Development

ISSN : 08537380 EISSN : 2252696X DOI : -

Core Subject : Health,

Veterinary

Aims JITV (Jurnal Ilmu ternak dan Veteriner) or Indonesian Journal of Animal and Veterinary Sciences (IJAVS) aims to publish original research results and reviews on farm tropical animals such as cattle, buffaloes, sheep, goats, pigs, horses, poultry, as well as non domesticated Indonesian endemic animals, such as deers, anoa, babirusa, etc. Scope Indonesian Journal of Animal and Veterinary Sciences . The journal will consider primary research papers from any source if they make an original contribution to the experimental or theoretical understanding and application of theories and methodologies of some aspects of animal science and veterinary

Arjuna Subject : Ilmu Pertanian dan Biologi (Semua) - Ilmu Hewan dan Zoologi

Articles 1,756 Documents

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An investigation of the feasibility of vaccinating against the Old World Screwworm fly Chrysomya bezziana Willadseni, Peter; Partoutomo, Sujitno
Indonesian Journal of Animal and Veterinary Sciences Vol 5, No 3 (2000)
Publisher : Indonesian Animal Sciences Society

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. Â Key words: Chrysomya bezziana, screwworm fly, vaccine

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

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. Â 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

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 Â 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 Â roteins as recombinants in a form suitable for vaccination studies. Â Key words: Chrysomya bezziana, peritrophic membrane, vaccination, amino acid sequence, serine protease

cDNA library construction and isolation of genes for candidate vaccine antigens from Chrysomya bezziana (the Old World Screwworm fly) Muharsini, Sri; Wijffe, Gene; Voucolo, Tony; Supriyanti, Florentina
Indonesian Journal of Animal and Veterinary Sciences Vol 5, No 3 (2000)
Publisher : Indonesian Animal Sciences Society

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 Î» ZAP expressÂ®. 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. Â Key words: cDNA library, peritrophin, peritrophic membrane, Chrysomya bezziana, Lucilia cuprina, vaccine

Bacterial expression of larval peritrophins of Chryosomya bezziana Wijffels, Gene; Voucoloco, Tony; Muharsini, Sri; Supriyanti, Florentina
Indonesian Journal of Animal and Veterinary Sciences Vol 5, No 3 (2000)
Publisher : Indonesian Animal Sciences Society

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 â42 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. Â Key words: Chrysomya, peritrophic membrane, peritrophin, recombinant protein, hexaHis tag, GST

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
Indonesian Journal of Animal and Veterinary Sciences Vol 5, No 3 (2000)
Publisher : Indonesian Animal Sciences Society

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. Â Key words: Chrysomya bezziana, peritrophin, Pichia pastoris

Purification of recombinant peritrophic membrane proteins of the Old World Screwworm fly, Chrysomya bezziana Pearson, Roger; Muharsini, Sri; Wijffels, Gene; Voucoloco, Tony
Indonesian Journal of Animal and Veterinary Sciences Vol 5, No 3 (2000)
Publisher : Indonesian Animal Sciences Society

To evaluate the feasibility of vaccinating sheep against the Old World Screwworm fly Chrysomya bezziana several recombinant peritrophin proteins were expressed in either a denatured form in Escherichia coli or a native-like form in Pichia pastoris cultures. Purification of the hexaHis tagged proteins was achieved by immobilized metal affinity chromatography. Proteins purified under reducing conditions were refolded using a glutathione shuffle procedure. Purification of a glutathione-Stransferase fusion protein was attempted using glutathione affinity chromatography in conjunction with anion exchange chromatography. The authenticity of the expressed proteins was verified by amino terminal amino acid sequencing. Carbohydrate analysis using biotinylated lectins revealed that Cb-peritrophin-48 expressed in Pichia pastoris was glycosylated with high mannose-type sugars. Four of the purified recombinant proteins were used to evaluate their protective immunogenicity in sheep against Chrysomya bezziana strike. Â Key words: Screwworm fly, Chrysomya bezziana, recombinant proteins, immobilized metal affinity chromatography

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
Indonesian Journal of Animal and Veterinary Sciences Vol 5, No 3 (2000)
Publisher : Indonesian Animal Sciences Society

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. Â Key words: Chrysomya bezziana, recombinant antigen, peritrophin, vaccination

Localization of the glycoprotein Cb42 in larvae of the screwworm fly Chrysomya bezziana (Diptera: Calliphoridae) Eisemanni, Creig; Muharsini, Sri
Indonesian Journal of Animal and Veterinary Sciences Vol 5, No 3 (2000)
Publisher : Indonesian Animal Sciences Society

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. Â Key words: Cb-peritrophin-42, Cb42, glycoprotein, peritrophic membrane, immunolocalization

Vaccination against Chrysomya bezziana: a summary of current knowledge Willadseni, Peter
Indonesian Journal of Animal and Veterinary Sciences Vol 5, No 3 (2000)
Publisher : Indonesian Animal Sciences Society

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. Â Key words: Chrysomya bezziana, Old World screwworm fly, vaccine, recombinant antigen, sheep

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Ahmadi Riyanto

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Home Page

OAI Link

Editorial Team

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Contact Name Ahmadi Riyanto

Contact Email medpub@litbang.deptan.go.id

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Location Kota bogor, Jawa barat INDONESIA

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Contact Name
Ahmadi Riyanto

Contact Email
medpub@litbang.deptan.go.id

Phone
-

Journal Mail Official
ahmadi_puslitbangnak@yahoo.com

Editorial Address
-

Location
Kota bogor,

Jawa barat

INDONESIA