DAO 86:253-262 (2009)  -  DOI: https://doi.org/10.3354/dao02121

Changes induced by two strains of Vibrio splendidus in haemocyte subpopulations of Mya arenaria, detected by flow cytometry with LysoTracker

Dante R. Mateo1,*, Aleks Spurmanis2, Ahmed Siah1, Mebrahtu T. Araya1, Mariana Kulka2, Franck C. J. Berthe1,3, Gerry R. Johnson1, Spencer J. Greenwood1,4

1Department of Pathology and Microbiology, Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue, Charlottetown, Prince Edward Island C1A 4P3, Canada
2Institute for Nutrisciences and Health, National Research Council (NRC), Regis and Joan Duffy Research Centre, University of Prince Edward Island, 550 University Avenue, Charlottetown, Prince Edward Island C1A 4P3, Canada
3Animal Health and Welfare Unit, European Food Safety Authority (EFSA), Largo N, Palli 5IA, 43100 Parma, Italy
4AVC Lobster Science Centre, Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue, Charlottetown, Prince Edward Island C1A 4P3, Canada

ABSTRACT: Flow-cytometric characterisation of bivalve haemocytes is usually performed by light-scatter profiles based on size and complexity of the cells. Additional means of characterisation such as specific fluorescent dyes are not commonly used to discriminate cell subpopulations in challenged and unchallenged haemocytes. In the present study, we characterise the changes in haemocyte subpopulations of soft-shell clam Mya arenaria induced by in vivo challenge with 2 strains of Vibrio splendidus by using a fluorescent probe. Responses were measured 24 h after infection with either a local wild strain (7SHRW) or a modification (LGP32-GFP) of a strain associated with oyster mortalities in France (LGP32). Changes in haemocyte subpopulations were analysed using flow cytometry based on 2-parameter scatter profiles and lysosomal content reflected by LysoTracker staining. Forward and side-scatter profiles revealed 2 haemocyte subpopulations: hyalinocytes and granulocytes. Granulocytes exhibited significantly higher levels of lysosomal staining (p < 0.01). Following infection with LGP32-GFP, both subpopulations merged into a single continuous group and their lysosomal content significantly decreased (p < 0.05). Independent modifications after infection were observed in the proportions of subpopulations established by their lysosomal content. While the subpopulation of hyalinocytes had lower levels of lysosomal content after infection, especially with LGP32-GFP (p < 0.001), the subpopulation of granulocytes had similar levels of lysosomes after infection with 7SHRW and significantly decreased levels after infection with LGP32-GFP (p = 0.001). Our data suggest specific modulation of bivalve responses against pathogenic bacteria that would include degranulation.


KEY WORDS: Mya arenaria · Vibrio splendidus · Haemocyte subpopulations · Flow cytometry · LysoTracker · Lysosome


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Cite this article as: Mateo DR, Spurmanis A, Siah A, Araya MT and others (2009) Changes induced by two strains of Vibrio splendidus in haemocyte subpopulations of Mya arenaria, detected by flow cytometry with LysoTracker. Dis Aquat Org 86:253-262. https://doi.org/10.3354/dao02121

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