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Journal of Shellfish Research. Vol. 25, NO.3, 1051-1057, 2006. FLUOROMETRIC MEASUREMENT OF OXIDATIVE BURST IN LOBSTER HEMOCYTES AND
 

Summary: Journal of Shellfish Research. Vol. 25, NO.3, 1051-1057, 2006.
FLUOROMETRIC MEASUREMENT OF OXIDATIVE BURST IN LOBSTER HEMOCYTES AND
INHIBITING EFFECT OF PATHOGENIC BACTERIA AND HYPOXIA
BRANDON MOSS AND BASSEM ALLAM*
Marine Sciences Research Center, Stony Brook University, Stony Brook, New York 11794
ABSTRACT The stimulation of hemocytes during phagocytosis leads to the generation of a series of oxygen radicals known as
reactive oxygen species (ROS).Among these, hydrogen peroxide plays an important microbicidal role by directly killing microor-
ganisms or by sewing as an intermediate for other antimicrobial radicals. In this study, we adapted a technique using 2',7'-
dichlorofluorescein-diacetate(DCFH-DA)to measure H202production in lobster hemocytes. After oxidation by hydrogen peroxide,
this molecule produces a fluorescent product that can be easily detected. The respiratory burst was successfully activated in lobster
hemocytes by the addition of zymosan particles,but not with phorbol myristate acetate. After optimization,we used the technique to
investigate the effect of different bacterial strains, including lobster pathogens, on the oxidative burst. Results demonstrate that
Aureococcus viridans, a bacterial pathogen that is able to survive phagocytosis by lobster hemocytes, quenches ROS production. The
comparisonof ROS production in lobsters collected from field sites submitted to different levels of dissolved oxygen suggests that this
technique provides a good indicator of lobster physiological status and immunocompetency.
KEY WOIZDS: lobster, Homarus americanus, oxygen, in v i m , dichlorofluorescein diacetate, ROS
INTRODUCTION
The American lobster, Homarus arnericanus,is one of the most
commercially important species in the Northeastern United States
and Canada. This species is subject to different infections, includ-

  

Source: Allam, Bassem - School of Marine and Atmospheric Sciences, SUNY at Stony Brook

 

Collections: Biology and Medicine; Environmental Sciences and Ecology