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Donaghy, Ludovic,Hong, Hyun-Ki,Kim, Moonkoo,Park, Heung-Sik,Choi, Kwang-Sik Elsevier 2016 MARINE POLLUTION BULLETIN Vol.113 No.1
<P><B>Abstract</B></P> <P>In December 2007, >150km of the West coast of Korea were heavily polluted by crude oil leaked from the oil tanker <I>Hebei Spirit</I>, leading to mass mortality of bivalve mollusks on the intertidal areas. Two years after, mussels <I>Mytilus galloprovincialis</I> were collected from two impacted sites to investigate sub-lethal effects of the oil spill. Tissue content in polycyclic aromatic hydrocarbons (PAHs), hemocyte parameters, reproductive status and energetic reserves were analyzed. PAHs in tissues of mussels as well as hemocyte parameters were not different between impacted and control sites. Energetic reserves were altered in mussels from the impacted sites. Glycogen content remained low at polluted sites, whatever the season. Two years after the <I>Hebei Spirit</I> oil spill, mussels then presented altered energetic metabolism. Further investigations are thus warranted to monitor the sustainability of mussel populations on the oil spilled West coast of Korea.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Fitness of mussels was assessed two years after the <I>Hebei spirit</I> oil spill. </LI> <LI> Hemocyte parameters are similar to control. </LI> <LI> Mussels present altered energetic metabolisms. </LI> <LI> Altered fitness of mussels may threaten the sustainability of the populations. </LI> </UL> </P>
Donaghy, L.,Hong, H.K.,Jauzein, C.,Choi, K.S. Academic Press 2015 FISH AND SHELLFISH IMMUNOLOGY Vol.42 No.1
Reactive oxygen and nitrogen species (ROS and RNS) are naturally produced in all cells and organisms. Modifications of standard conditions alter reactive species generation and may result in oxidative stress. Because of the degradation of marine ecosystems, massive aquaculture productions, global change and pathogenic infections, oxidative stress is highly prevalent in marine bivalve molluscs. Haemocytes of bivalve molluscs produce ROS and RNS as part of their basal metabolism as well as in response to endogenous and exogenous stimuli. However, sources and pathways of reactive species production are currently poorly deciphered in marine bivalves, potentially leading to misinterpretations. Although sources and pathways of ROS and RNS productions are highly conserved between vertebrates and invertebrates, some uncommon pathways seem to only exist in marine bivalves. To understand the biology and pathobiology of ROS and RNS in haemocytes of marine bivalves, it is necessary to characterise their sources and pathways of production. The aims of the present review are to discuss the currently known and unknown intracellular sources of reactive oxygen and nitrogen species in marine bivalve molluscs, in light of terrestrial vertebrates, and to expose principal pitfalls usually encountered.
Donaghy, L.,Kim, B.K.,Hong, H.K.,Park, H.S.,Choi, K.S. Academic Press 2009 FISH AND SHELLFISH IMMUNOLOGY Vol.27 No.2
The Suminoe oyster Crassostrea ariakensis has been attempted to be introduced in the Chesapeake Bay, USA, as an alternative to the eastern oyster Crassostrea virginica. Commercial production of Suminoe oysters is currently restricted due to the incomplete understanding of their biological, physiological and immunological nature. Accordingly, understanding immune system of C. ariakensis is crucial to prevent disease associated mortality and subsequent management of the Suminoe oyster. We investigated immunological activities and morphology of hemocytes of the Suminoe oyster using flow cytometry and light microscopy. Three types of hemocytes were identified in the hemolymph including hyalinocyte, granulocyte and blast-like cells. Hyalinocytes were the largest cells and the most abundant, while granulocytes were intermediate-size cell containing numerous granules in the cytoplasm. Blast-like cells were the smallest and least numerous. Flow cytometry revealed that the granulocytes are most active in the cell phagocytosis and spontaneous reactive oxygen species (ROS) production. The hyalinocytes also showed a certain level of the phagocytosis and oxidative activity but in a lesser extent than the granulocytes. In contrast, the blast-like cells did not show any phagocytosis or oxidative activity. The flow cytometry used in this study confirmed that as observed from other marine bivalves, the granulocytes are the main hemocytes involved in the cellular defence in the Suminoe oyster.
Donaghy, L.,Hong, H.K.,Lambert, C.,Park, H.S.,Shim, W.J.,Choi, K.S. Academic Press 2010 FISH AND SHELLFISH IMMUNOLOGY Vol.28 No.1
The disk abalone Haliotis discus discus and the spiny top shell Turbo cornutus are edible gastropod species of high economic value, mainly in Asia. Mortality outbreaks and variations in worldwide stock abundance have been reported and suggested to be associated, at least in part, with pathogenic infections. Ecology, biology and immunology of both species are currently not well documented. The characterisation of the immune systems of these species is necessary to further assess the responses of H. discus discus and T. cornutus to environmental, chemical and disease stresses. In the present study, we investigated the morphology and immune-related activities of hemocytes in both species using light microscopy and flow cytometry. Two types of hemocytes were identified in the disk abalone hemolymph, blast-like cells and hyalinocytes; whereas four main hemocyte types were distinguished in the spiny top shell, blast-like cells, type I and II hyalinocytes, and granulocytes. Flow cytometric analysis also revealed differences between cell types in immune-related activities. Three subsets of hemocytes, defined by differing lysosomal characteristics, were observed in the hemolymph of the spiny top shell, and only one in the disk abalone. Phagocytic activity was higher in H. discus discus hemocytes than in T. cornutus hemocytes, and the kinetics of PMA-stimulated oxidative activity was different between hemocytes of the disk abalone and the spiny top shell. Finally our results suggest for the first time a predominant mitochondrial origin of oxidative activity in gastropod hemocytes.
Flow cytometric characterization of hemocytes of the solitary ascidian, <i>Halocynthia roretzi</i>
Donaghy, Ludovic,Hong, Hyun-Ki,Park, Kyung-Il,Nobuhisa, Kajino,Youn, Seok-Hyun,Kang, Chang-Keun,Choi, Kwang-Sik Elsevier 2017 FISH AND SHELLFISH IMMUNOLOGY Vol.66 No.-
<P><B>Abstract</B></P> <P>Internal defense of ascidians relies, at least partially, on cells circulating in body fluids and infiltrating in tissues, referred to as hemocytes, although structure and composition of ascidian hemocytes still remain unclear. In the present study, we investigated hemocyte types and their functions of the solitary ascidian <I>Halocynthia roretzi</I> using flow cytometry. Based on morphology, cellular activities and intracellular parameters from the flow cytometry, we identified eight hemocyte types including, three granulocytes (Gr-1, Gr-2, and Gr-3), 4 hyalinocytes (Hy-1, Hy-1′, Hy-2, and Hy-3) and lymphocyte-like (Ly-like) cells. The granulocyte Gr-1 accounted for 30% of the total circulating hemocytes and exhibited highest density of lysosomes and oxidative activity. Gr-1 was deeply involved in phagocytosis and degradation of foreign material. Hyalinocytes consist of two main populations, Hy-1 and Hy-2, and each accounted for 30% of the circulating hemocyte. Hy-1 displayed lysosomal content, an inducible oxidative activity, and no proteases, while Hy-2 expressed highest density of intracellular proteases, no lysosomes and a low oxidative activity. It was believed that Hy-2 may represent an important link between cellular and humoral immune reactions. Hy-1 did not show phagocytosis activity. Hy-3 and the Ly-like cells presented a similar profile except for their size and complexity, and Hy-3 may represent an intermediate differentiation/maturation step between Ly-like cells and other hemocyte populations. This first characterization of the hemocyte populations of <I>H. roretzi</I> provides a solid basis to investigate further their respective roles and functions in physiological and pathological contexts.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A few studies have investigated structure and types of ascidian immune cells. </LI> <LI> We characterized hemocytes of the ascidian <I>Halocynthia roretzi</I> using flow cytometry. </LI> <LI> Eight types of hemocytes identified: 3 granulocytes, 4 hyalinocytes and lymphocyte-like cells. </LI> <LI> This study provides a basis to investigate further respective roles and functions of the ascidian hemocytes. </LI> </UL> </P>
박경일,Ludovic Donaghy,강현실,홍현기,김영옥,최광식 한국해양과학기술원 2012 Ocean science journal Vol.47 No.1
Cellular and humoral immune parameters are often used as biomarkers to trace environmental and physiological stresses in marine bivalves. In this study, we compared various immune parameters of Manila clams (Ruditapes philippinarum) under normal conditions and under a high level of desiccation,using flow cytometry. The immune parameters analyzed included, total hemocyte count, hemocyte mortality, hemocyte DNA damage, reactive oxygen species (ROS) production, and phagocytosis activity. Total hemocyte count, hemocyte DNA damage, and hemocyte mortality were significantly elevated among clams under high desiccation stress, while phagocytosis activity and spontaneous ROS production were significantly lower compared to those parameters of the control clams (p<0.05). These data suggest that the immune parameters analyzed in this study well reflect the physiological status of clams.