Flow cytometric analysis of haemocytes from eastern oysters, Crassostrea virginica, subjected to a sudden temperature elevation

Hégaret, Hélène; Wikfors, Gary H.; Soudant, Philippe

doi:10.1016/s0022-0981(03)00235-1

Cited by 241 publications

(127 citation statements)

References 25 publications

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“…Hemocytes from many bivalve species are known to be able to produce ROS. Oxidative mechanisms were reported in the oysters C. virginica, C. gigas, C. ariakensis and O. edulis (Larson et al, 1989;Bachère et al, 1991;Hégaret et al, 2003b;Lambert et al, 2003;Goedken and DeGuise, 2004;Labreuche et al, 2006;Donaghy et al, 2009), the mussels M. edulis and M. galloprovincialis (Pipe, 1992;Carballal et al, 1997b;Ordás et al, 2000b) and the scallops P. maximus and C. farreri (Le Gall et al, 1991;Chen et al, 2007a,b). In clams, few data are currently available and contradictory results between species were reported.…”

Section: Oxidative Mechanismsmentioning

confidence: 99%

Hemocytes of the carpet shell clam (Ruditapes decussatus) and the Manila clam (Ruditapes philippinarum): Current knowledge and future prospects

et al. 2009

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: Hemocytes are the immune effectors in clams, directing cellular but also humoral defences. In the Manila clam (Ruditapes philippinarum) and the carpet shell clam (Ruditapes decussatus), two phenotypically similar venerid species, fishery and aquaculture importance motivated scientific studies to improve knowledge about the participation of hemocytes in immune response. Currently, gaps still persist concerning the classification of hemocyte sub-populations as well as their non-immune activities. This review summarizes current knowledge about hemocytes of the Manila clam and the carpet shell clam, suggests some probable misinterpretations arising from extrapolations from other bivalves and vertebrates, and suggests future research prospects. The susceptibility of hemocytes to environmental modifications as well as biological and chemical toxicants is also re-evaluated. Hemocytes of bivalves are not limited in function to immunity and should be considered in a wider scheme including the environment and whole-animal physiology.

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Section: Oxidative Mechanismsmentioning

confidence: 99%

Hemocytes of the carpet shell clam (Ruditapes decussatus) and the Manila clam (Ruditapes philippinarum): Current knowledge and future prospects

et al. 2009

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“…Hemato-immunological parameters measured were: the concentration of circulating hemocytes (=total hemocyte counts À THC, cells ml À1 ) as well as hemocyte characterization, in terms of size using the FSC detector and granularity or internal complexity using the SSC detector, according to Hégaret et al (2003a). The immune functions measured were:…”

Section: Analysis Of Hemocyte Parametersmentioning

confidence: 99%

“…according to Hégaret et al (2003b); (b) Phagocytosis of fluorescent microbeads (Fluoresbrite YG Microspheres, 2.00 lm, Polysciences) by hemocytes, as percentage of highly-phagocytic (>2 beads) hemocytes according to Hégaret et al (2003b) and by the number of beads phagocytized by each hemocyte (G mean(>2 beads) /G mean(1 bead) ); (c) Hemocyte production of reactive oxygen species (ROS) with potential to kill non-self, engulfed particles was assessed using 2 V,7 V-dichlorofluorescein diacetate (DCFH-DA, Sigma) described in Buggé et al (2007). (d) Adhesion of the hemocytes was measured by assessing the proportion of hemocytes that detach from the surface of experimental chambers after incubation with potential toxins as previously described for clams by Choquet et al (2003).…”

Section: Analysis Of Hemocyte Parametersmentioning

confidence: 99%

Combined effects of a parasite, QPX, and the harmful-alga, Prorocentrum minimum on northern quahogs, Mercenaria mercenaria

Hégaret

Smolowitz

Sunila

et al. 2010

Marine Environmental Research

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a b s t r a c tNorthern quahogs, Mercenaria mercenaria (L.), frequently are infected with the parasite Quahog Parasite Unknown (QPX, Labyrintohomorpha, Thraustochytriales), which can cause morbidity and mortality of the quahogs. Possible interactions between this parasitic disease and exposure to the harmful dinoflagellate Prorocentrum minimum in M. mercenaria were studied experimentally. Quahogs from Massachusetts with variable intensity of QPX infection were exposed, under controlled laboratory conditions, to cultured P. minimum added to the natural plankton at a cell density equivalent to a natural bloom. After 5 days of exposure, individual clams were diagnosed histologically to assess prevalence and intensity of parasitic infection, as well as other pathological conditions. Further, cellular defense status of clams was evaluated by analyzing hemocyte parameters (morphological and functional) using flow-cytometry. Exposure of quahogs to P. minimum resulted in: a lower percentage of phagocytic hemocytes, higher production of reactive oxygen species (ROS), larger hemocyte size, more-numerous hemocytic aggregates, and increased numbers of hemocytes in gills accompanied by vacuolation and hyperplasia of the water-tubular epithelial cells of the gills. Quahogs had a low prevalence of QPX; by chance, the parasite was present only in quahogs exposed to P. minimum. Thus, the effect of QPX alone on the hemocyte parameters of quahogs could not be assessed in this experiment, but it was possible to assess different responses of infected versus non-infected quahogs to P. minimum. QPX-infected quahogs exposed to P. minimum had repressed percentage of phagocytic hemocytes, consistent with immuno-modulating effect of P. minimum upon several molluscan species, as well as smaller hemocytes and increased hemocyte infiltration throughout the soft tissues. This experiment demonstrates the importance of considering interactive effects of different factors on the immunology and histopathology of bivalve shellfish, and highlights the importance of considering the presence of parasites when bivalves are subjected to harmful-algal blooms.

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“…Calcein (or other similar cell-permeant fluorophores) and PI (or other similar cell-impermeant fluorophores) have long been used together as 'live/dead stains' in mammalian cells in vitro (Dive et al, 1990) but these have rarely been used together in marine invertebrates. However, hemocyte viability has been assessed in bivalves and crustaceans using calcein alone (Cardenas et al, 2004;Marin et al, 2004) and PI alone (Hegaret et al, 2003;Lee et al, 2001;Xue et al, 2001), and in G. dibranchiata erythrocytes in vivo using PI alone (Hance et al, 2008). We found that under control conditions in vitro, 100% of G. dibranchiata erythrocytes were labeled by calcein but were not labeled by PI whereas at 3% H 2 S (approximately 5 mmol l -1 dissolved sulfide), 99% of the cells were labeled by PI but were not labeled by calcein.…”

Section: J a Ortega J M Ortega And D Julianmentioning

confidence: 99%

Hypotaurine and sulfhydryl-containing antioxidants reduce H2S toxicity in erythrocytes from a marine invertebrate

Ortega

Julian

2008

Journal of Experimental Biology

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SUMMARYHypotaurine (HT) has been proposed to reduce sulfide toxicity in some deep-sea invertebrates by scavenging free radicals produced from sulfide oxidation or by scavenging sulfide via the reaction of HT with sulfide, forming thiotaurine (ThT). We tested whether HT or several antioxidants could reduce the total dissolved sulfide concentration in buffered seawater exposed to H 2 S, and whether HT, ThT or antioxidants could increase the viability of Glycera dibranchiata erythrocytes exposed to H 2 S in vitro. We found that 5 and 50 mmol l -1 HT reduced the dissolved sulfide in cell-free buffer exposed to H 2 S by up to 80% whereas the antioxidants glutathione ethyl ester (GEE) ), which contain sulfhydryl groups, increased cell viability during H 2 S exposure but to a lesser extent than HT whereas ASC, Tempol and Trolox, which do not contain sulfhydryl groups, decreased viability or had no effect. These data show that HT can protect cells from sulfide in vitro and suggest that sulfide scavenging, rather than free radical scavenging, is the most important mechanism of protection.,

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Flow cytometric analysis of haemocytes from eastern oysters, Crassostrea virginica, subjected to a sudden temperature elevation

Cited by 241 publications

References 25 publications

Hemocytes of the carpet shell clam (Ruditapes decussatus) and the Manila clam (Ruditapes philippinarum): Current knowledge and future prospects

Hemocytes of the carpet shell clam (Ruditapes decussatus) and the Manila clam (Ruditapes philippinarum): Current knowledge and future prospects

Combined effects of a parasite, QPX, and the harmful-alga, Prorocentrum minimum on northern quahogs, Mercenaria mercenaria

Hypotaurine and sulfhydryl-containing antioxidants reduce H2S toxicity in erythrocytes from a marine invertebrate

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