Generation of Paramoeba perurans clonal cultures using flow cytometry and confirmation of virulence

C Collins, M Hall, D Bruno, J Sokolowska, L Duncan, R Yuecel, U McCarthy, M J Fordyce, C C Pert, R McIntosh, Z MacKay

Research output: Contribution to journalArticle

8 Citations (Scopus)
4 Downloads (Pure)

Abstract

Amoebic gill disease (AGD) in farmed Atlantic salmon is caused by the amoeba Paramoeba perurans. The recent establishment of in vitro culture techniques for P. perurans has provided a valuable tool for studying the parasite in detail. In this study, flow cytometry was used to generate clonal cultures from single-sorted amoeba, and these were used to successfully establish AGD in experimental Atlantic salmon. The clonal cultures displayed differences in virulence, based on gill scores. The P. perurans load on gills, determined by qPCR analysis, showed a positive relationship with gill score, and with clonal virulence, indicating that the ability of amoebae to proliferate and/or remain attached on gills may play a role in virulence. Gill scores based on gross signs and histopathological analysis were in agreement. No association between level of gill score and specific gill arch was observed. It was found that for fish with lower gill scores based on histopathological examination, gross examination and qPCR analysis of gills from the same fish were less successful in detecting lesions and amoebae, respectively.
Original languageEnglish
Pages (from-to)351–365
Number of pages15
JournalJournal of Fish Diseases
Volume40
Issue number3
Early online date15 Aug 2016
DOIs
Publication statusPublished - 1 Mar 2017

Keywords

  • Paramoeba
  • perurans
  • flow cytometry
  • clonal
  • culture
  • virulence

Fingerprint Dive into the research topics of 'Generation of <i>Paramoeba perurans</i> clonal cultures using flow cytometry and confirmation of virulence'. Together they form a unique fingerprint.

  • Equipment

  • BD LSR Fortessa

    Andrea Holme (Manager)

    Iain Fraser Cytometry Centre

    Research Facilities: Equipment

  • Iain Fraser Cytometry Centre

    Andrea Holme (Manager), Linda Duncan (Senior Application Scientist), Ailsa Laird (Technician) & Kate Burgoyne (Technician)

    Institute of Medical Sciences

    Research Facilities: Facility

  • Cite this

    Collins, C., Hall, M., Bruno, D., Sokolowska, J., Duncan, L., Yuecel, R., McCarthy, U., Fordyce, M. J., Pert, C. C., McIntosh, R., & MacKay, Z. (2017). Generation of Paramoeba perurans clonal cultures using flow cytometry and confirmation of virulence. Journal of Fish Diseases, 40(3), 351–365. https://doi.org/10.1111/jfd.12517