A putative serine protease, SpSsp1, from Saprolegnia parasitica is recognised by sera of rainbow trout, Oncorhynchus mykiss

Kirsty L. Minor, Victoria L. Anderson, Katie S. Davis, Albert H. Van Den Berg, James S. Christie, Lars Loebach, Ali Reza Faruk, Stephan Wawra, Chris J. Secombes, Pieter Van West

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Abstract

Saprolegniosis, the disease caused by Saprolegnia sp., results in considerable economic losses in aquaculture. Current control methods are inadequate, as they are either largely ineffective or present environmental and fish health concerns. Vaccination of fish presents an attractive alternative to these control methods. Therefore we set out to identify suitable antigens that could help generate a fish vaccine against Saprolegnia parasitica. Unexpectedly, antibodies against S. parasitica were found in serum from healthy rainbow trout, Oncorhynchus mykiss. The antibodies detected a single band in secreted proteins that were run on a one-dimensional SDS-polyacrylamide gel, which corresponded to two protein spots on a two-dimensional gel. The proteins were analysed by liquid chromatography tandem mass spectrometry. Mascot and bioinformatic analysis resulted in the identification of a single secreted protein, SpSsp1, of 481 amino acid residues, containing a subtilisin domain. Expression analysis demonstrated that SpSsp1 is highly expressed in all tested mycelial stages of S. parasitica. Investigation of other non-infected trout from several fish farms in the United Kingdom showed similar activity in their sera towards SpSsp1. Several fish that had no visible saprolegniosis showed an antibody response towards SpSsp1 suggesting that SpSsp1 might be a useful candidate for future vaccination trial experiments. (C) 2014 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Original languageEnglish
Pages (from-to)630-639
Number of pages10
JournalFungal Biology
Volume118
Issue number7
Early online date30 Apr 2014
DOIs
Publication statusPublished - Jul 2014

Keywords

  • Oncorhynchus mykiss
  • Pathogenicity factor
  • Saprolegnia parasitica
  • Serine protease
  • salmo-trutta
  • brown trout
  • Atlantic salmon
  • malachite green
  • gene-expression
  • fish
  • pathogenicity
  • mortality
  • database
  • water

Cite this

A putative serine protease, SpSsp1, from Saprolegnia parasitica is recognised by sera of rainbow trout, Oncorhynchus mykiss. / Minor, Kirsty L.; Anderson, Victoria L.; Davis, Katie S.; Van Den Berg, Albert H.; Christie, James S.; Loebach, Lars; Faruk, Ali Reza; Wawra, Stephan; Secombes, Chris J.; Van West, Pieter.

In: Fungal Biology, Vol. 118, No. 7, 07.2014, p. 630-639.

Research output: Contribution to journalArticle

Minor, KL, Anderson, VL, Davis, KS, Van Den Berg, AH, Christie, JS, Loebach, L, Faruk, AR, Wawra, S, Secombes, CJ & Van West, P 2014, 'A putative serine protease, SpSsp1, from Saprolegnia parasitica is recognised by sera of rainbow trout, Oncorhynchus mykiss' Fungal Biology, vol. 118, no. 7, pp. 630-639. https://doi.org/10.1016/j.funbio.2014.04.008
Minor, Kirsty L. ; Anderson, Victoria L. ; Davis, Katie S. ; Van Den Berg, Albert H. ; Christie, James S. ; Loebach, Lars ; Faruk, Ali Reza ; Wawra, Stephan ; Secombes, Chris J. ; Van West, Pieter. / A putative serine protease, SpSsp1, from Saprolegnia parasitica is recognised by sera of rainbow trout, Oncorhynchus mykiss. In: Fungal Biology. 2014 ; Vol. 118, No. 7. pp. 630-639.
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title = "A putative serine protease, SpSsp1, from Saprolegnia parasitica is recognised by sera of rainbow trout, Oncorhynchus mykiss",
abstract = "Saprolegniosis, the disease caused by Saprolegnia sp., results in considerable economic losses in aquaculture. Current control methods are inadequate, as they are either largely ineffective or present environmental and fish health concerns. Vaccination of fish presents an attractive alternative to these control methods. Therefore we set out to identify suitable antigens that could help generate a fish vaccine against Saprolegnia parasitica. Unexpectedly, antibodies against S. parasitica were found in serum from healthy rainbow trout, Oncorhynchus mykiss. The antibodies detected a single band in secreted proteins that were run on a one-dimensional SDS-polyacrylamide gel, which corresponded to two protein spots on a two-dimensional gel. The proteins were analysed by liquid chromatography tandem mass spectrometry. Mascot and bioinformatic analysis resulted in the identification of a single secreted protein, SpSsp1, of 481 amino acid residues, containing a subtilisin domain. Expression analysis demonstrated that SpSsp1 is highly expressed in all tested mycelial stages of S. parasitica. Investigation of other non-infected trout from several fish farms in the United Kingdom showed similar activity in their sera towards SpSsp1. Several fish that had no visible saprolegniosis showed an antibody response towards SpSsp1 suggesting that SpSsp1 might be a useful candidate for future vaccination trial experiments. (C) 2014 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.",
keywords = "Oncorhynchus mykiss, Pathogenicity factor, Saprolegnia parasitica, Serine protease, salmo-trutta, brown trout, Atlantic salmon, malachite green, gene-expression, fish, pathogenicity, mortality, database, water",
author = "Minor, {Kirsty L.} and Anderson, {Victoria L.} and Davis, {Katie S.} and {Van Den Berg}, {Albert H.} and Christie, {James S.} and Lars Loebach and Faruk, {Ali Reza} and Stephan Wawra and Secombes, {Chris J.} and {Van West}, Pieter",
note = "Acknowledgements Our work was supported by the BBSRC (BB/C518457/1, BB/G012075/1, BB/J018333/1) (K.L.M., C.J.S., J.S.C., K.S.D., and P.v.W.), the University of Aberdeen (V.L.A., C.J.S., and P.v.W.), MSD Animal Health (J.S.C., K.S.D., and A.H.v.d.B), and The Royal Society (P.v.W.). This work was also supported by a Marie Curie Initial Training Networks with the SAPRO (sustainable approaches to reduce Oomycete (Saprolegnia) infections in aquacultures) grant PITN-GA-2009-238550 (A.H.v.d.B., L.L., C.J.S., P.v.W.). We would like to acknowledge Aberdeen Proteomics for carrying out LC–MS/MS and Laura Grenville-Briggs for valuable discussion and technical help. We are grateful to the Broad Institute (Carsten Russ, Rays Jiang, Brian Haas, and Chad Nusbaum), Brett Tyler (VBI), and P.v.W. for early release of draft supercontigs of the genome sequence of isolate CBS233.65, which helped us identify SpSsp1.",
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TY - JOUR

T1 - A putative serine protease, SpSsp1, from Saprolegnia parasitica is recognised by sera of rainbow trout, Oncorhynchus mykiss

AU - Minor, Kirsty L.

AU - Anderson, Victoria L.

AU - Davis, Katie S.

AU - Van Den Berg, Albert H.

AU - Christie, James S.

AU - Loebach, Lars

AU - Faruk, Ali Reza

AU - Wawra, Stephan

AU - Secombes, Chris J.

AU - Van West, Pieter

N1 - Acknowledgements Our work was supported by the BBSRC (BB/C518457/1, BB/G012075/1, BB/J018333/1) (K.L.M., C.J.S., J.S.C., K.S.D., and P.v.W.), the University of Aberdeen (V.L.A., C.J.S., and P.v.W.), MSD Animal Health (J.S.C., K.S.D., and A.H.v.d.B), and The Royal Society (P.v.W.). This work was also supported by a Marie Curie Initial Training Networks with the SAPRO (sustainable approaches to reduce Oomycete (Saprolegnia) infections in aquacultures) grant PITN-GA-2009-238550 (A.H.v.d.B., L.L., C.J.S., P.v.W.). We would like to acknowledge Aberdeen Proteomics for carrying out LC–MS/MS and Laura Grenville-Briggs for valuable discussion and technical help. We are grateful to the Broad Institute (Carsten Russ, Rays Jiang, Brian Haas, and Chad Nusbaum), Brett Tyler (VBI), and P.v.W. for early release of draft supercontigs of the genome sequence of isolate CBS233.65, which helped us identify SpSsp1.

PY - 2014/7

Y1 - 2014/7

N2 - Saprolegniosis, the disease caused by Saprolegnia sp., results in considerable economic losses in aquaculture. Current control methods are inadequate, as they are either largely ineffective or present environmental and fish health concerns. Vaccination of fish presents an attractive alternative to these control methods. Therefore we set out to identify suitable antigens that could help generate a fish vaccine against Saprolegnia parasitica. Unexpectedly, antibodies against S. parasitica were found in serum from healthy rainbow trout, Oncorhynchus mykiss. The antibodies detected a single band in secreted proteins that were run on a one-dimensional SDS-polyacrylamide gel, which corresponded to two protein spots on a two-dimensional gel. The proteins were analysed by liquid chromatography tandem mass spectrometry. Mascot and bioinformatic analysis resulted in the identification of a single secreted protein, SpSsp1, of 481 amino acid residues, containing a subtilisin domain. Expression analysis demonstrated that SpSsp1 is highly expressed in all tested mycelial stages of S. parasitica. Investigation of other non-infected trout from several fish farms in the United Kingdom showed similar activity in their sera towards SpSsp1. Several fish that had no visible saprolegniosis showed an antibody response towards SpSsp1 suggesting that SpSsp1 might be a useful candidate for future vaccination trial experiments. (C) 2014 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

AB - Saprolegniosis, the disease caused by Saprolegnia sp., results in considerable economic losses in aquaculture. Current control methods are inadequate, as they are either largely ineffective or present environmental and fish health concerns. Vaccination of fish presents an attractive alternative to these control methods. Therefore we set out to identify suitable antigens that could help generate a fish vaccine against Saprolegnia parasitica. Unexpectedly, antibodies against S. parasitica were found in serum from healthy rainbow trout, Oncorhynchus mykiss. The antibodies detected a single band in secreted proteins that were run on a one-dimensional SDS-polyacrylamide gel, which corresponded to two protein spots on a two-dimensional gel. The proteins were analysed by liquid chromatography tandem mass spectrometry. Mascot and bioinformatic analysis resulted in the identification of a single secreted protein, SpSsp1, of 481 amino acid residues, containing a subtilisin domain. Expression analysis demonstrated that SpSsp1 is highly expressed in all tested mycelial stages of S. parasitica. Investigation of other non-infected trout from several fish farms in the United Kingdom showed similar activity in their sera towards SpSsp1. Several fish that had no visible saprolegniosis showed an antibody response towards SpSsp1 suggesting that SpSsp1 might be a useful candidate for future vaccination trial experiments. (C) 2014 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

KW - Oncorhynchus mykiss

KW - Pathogenicity factor

KW - Saprolegnia parasitica

KW - Serine protease

KW - salmo-trutta

KW - brown trout

KW - Atlantic salmon

KW - malachite green

KW - gene-expression

KW - fish

KW - pathogenicity

KW - mortality

KW - database

KW - water

U2 - 10.1016/j.funbio.2014.04.008

DO - 10.1016/j.funbio.2014.04.008

M3 - Article

VL - 118

SP - 630

EP - 639

JO - Fungal Biology

JF - Fungal Biology

SN - 1878-6146

IS - 7

ER -