In vitro effect of temperature on phagocytic activity in rainbow trout (Oncorhynchus mykiss)

Kód výsledku v IS VaVaI

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Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

In vitro effect of temperature on phagocytic activity in rainbow trout (Oncorhynchus mykiss)

Popis výsledku v původním jazyce

19th International Conference on Diseases of Fish and Shellfish, Porto, Portugal, 9. - 12. 9. 2019 – poster, abstract. Introduction: Phagocytosis assay represents a useful method for assessing the fish immune system and its ability to react to stimulants. During this assay, peripheral blood cells are incubated with various particles (e.g. zymosan, latex beads) in conditions specific for each fish species. Leukocytes are subsequently isolated by haemolysis in a hypotonic environment or gradient centrifugation. Measurement by flow cytometry is very effective and allows us to differentiate individual cell populations and easily evaluate the intensity of phagocytic activity. In fish, phagocytes comprise neutrophils, monocytes and also a small proportion of lymphocytes. Methodology varies between different authors, and there is a high natural variability between individual fish that needs to be considered. In this study, the optimal incubation temperature for rainbow trout phagocytes was investigated. Methodology: Peripheral blood was collected from the tail vein of rainbow trout (Oncorhynchus mykiss) into a heparinized syringe. Sample manipulation and incubation were performed at three different temperatures – 4°C, 15°C and 22°C (37°C). Leukocytes were incubated with various particles (zymosan Texas Red, Alexa Fluor 488) and isolated by haemolysis in a hypotonic environment. Isolated cells were centrifuged and fixed with EDTA. Differences between phagocyte numbers and the intensity of phagocytic activity at different temperatures were evaluated by flow cytometry. Results: The highest viability of phagocytic cells was obtained with the incubation at 4°C. However, phagocytic activity was suppressed at this temperature. Cell viability was further decreasing with rising temperature. The highest levels of phagocytic activity were recorded with the room temperature incubation (i.e. higher than the optimum temperature for rainbow trout). At 37°C, cell viability as well as phagocytic activity were decreased. Conclusion: Room temperature provided optimal conditions for in vitro phagocytosis of rainbow trout peripheral blood cells. However, the phagocytes viability was reduced compared to the incubation at lower temperatures; the assay therefore requires fast processing.

Název v anglickém jazyce

In vitro effect of temperature on phagocytic activity in rainbow trout (Oncorhynchus mykiss)

Popis výsledku anglicky

19th International Conference on Diseases of Fish and Shellfish, Porto, Portugal, 9. - 12. 9. 2019 – poster, abstract. Introduction: Phagocytosis assay represents a useful method for assessing the fish immune system and its ability to react to stimulants. During this assay, peripheral blood cells are incubated with various particles (e.g. zymosan, latex beads) in conditions specific for each fish species. Leukocytes are subsequently isolated by haemolysis in a hypotonic environment or gradient centrifugation. Measurement by flow cytometry is very effective and allows us to differentiate individual cell populations and easily evaluate the intensity of phagocytic activity. In fish, phagocytes comprise neutrophils, monocytes and also a small proportion of lymphocytes. Methodology varies between different authors, and there is a high natural variability between individual fish that needs to be considered. In this study, the optimal incubation temperature for rainbow trout phagocytes was investigated. Methodology: Peripheral blood was collected from the tail vein of rainbow trout (Oncorhynchus mykiss) into a heparinized syringe. Sample manipulation and incubation were performed at three different temperatures – 4°C, 15°C and 22°C (37°C). Leukocytes were incubated with various particles (zymosan Texas Red, Alexa Fluor 488) and isolated by haemolysis in a hypotonic environment. Isolated cells were centrifuged and fixed with EDTA. Differences between phagocyte numbers and the intensity of phagocytic activity at different temperatures were evaluated by flow cytometry. Results: The highest viability of phagocytic cells was obtained with the incubation at 4°C. However, phagocytic activity was suppressed at this temperature. Cell viability was further decreasing with rising temperature. The highest levels of phagocytic activity were recorded with the room temperature incubation (i.e. higher than the optimum temperature for rainbow trout). At 37°C, cell viability as well as phagocytic activity were decreased. Conclusion: Room temperature provided optimal conditions for in vitro phagocytosis of rainbow trout peripheral blood cells. However, the phagocytes viability was reduced compared to the incubation at lower temperatures; the assay therefore requires fast processing.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

40301 - Veterinary science

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů