Characterization of haemocytes of the Indian edible oyster, Crassostrea madrasensis (Preston) (original) (raw)
Related papers
Maintenance of haemocytes of edible oyster, Crassostrea madrasensis (Preston) in artificial media
Indian Journal of Fisheries, 2001
The haemocytes of Crassostrea madrasensis were maintained in Hank's balanced salt solution (HBSS), HBSS with 10% foetal calf serum (FCS), tissue culture media M 199, and M 199 with 10% FCS. Total counts of viable haemocytes were taken at 0 hr, 24 hrs, 48 hrs, and 72 hrs. HBSS and HBSS with 10% FCS were found to maintain cell viability upto 72 hrs without much reduction in the count of viable cells. In M199 and M199 with 10% FCS, it was found that a substantial increase of small hyalinocytes occurred at 24 hrs and 48 hrs. This was followed by a steep fall in the number of viable cells at 72 hrs in both the media. The HBSS, HBSS with 10% FCS, M199 and M199 with 10% FCS were found to be good for maintaining the cells in viable state. However, only M 199 and M 199 with 10% FCS indicated multiplication of haemocytes.
Sydney rock oyster (Saccostrea glomerata) hemocytes: Morphology and function
Journal of Invertebrate Pathology, 2007
In this study, three major hemocyte types were identified in the Sydney rock oyster. They were characterized primarily by light and electron microscopy based on the presence or absence of granules and nucleus to cytoplasm ratios. Hemoblast-like cells were the smallest cell type 4.0 ± 0.4 lm and comprised 15 ± 3% of the hemocyte population. They had large nuclei and scanty basic cytoplasm. This cell type also had some endoplasmic reticuli and mitochondria. The second major type were hyalinocytes. Hyalinocytes represented 46 ± 6% of all hemocytes. They were large cells (7.1 ± 1.0 lm) that had low nucleus:cytoplasm ratios and agranular basic or acidic cytoplasm. Hyalinocytes had the ability to phagocytose yeast cells and formed the core of hemocyte aggregates associated with agglutination. Four discrete sub-populations of hyalinocytes were identified. The third major cell type were the granulocytes, comprising 38 ± 1% of the hemocyte population. These cells were large (9.3 ± 0.3 lm) and were characterized by cytoplasm containing many acidic or basic granules. Granulocytes were more phagocytic than hyalinocytes and they formed the inner layer of hemocytes during the encapsulation of fungal hyphae. Five discrete sub-populations of granulocytes were identified based on the types of granules in their cytoplasm. Flow cytometry showed that the hemocytes of rock oysters could be divided into between two and four major cell types based on their light scattering properties. The most common of the cell types identified by flow cytometry corresponded to hyalinocytes and granulocytes. Cytochemical assays showed that most enzymes associated with immunological activity were localized in granulocytes. Their granules contained acid phosphatase, peroxidase, phenoloxidase, superoxide and melanin. Hyalinocytes were positive only for acid phosphatase. All of these observations suggest that Sydney rock oysters have a broad variety of functionally specialized hemocytes, many of which are involved in host defense.
Haemocyte morphology and function in the Akoya Pearl Oyster, Pinctada imbricata
Journal of Invertebrate Pathology, 2010
The morphology and cytochemistry of Pinctada imbricata haemocytes were studied in vitro. Three distinct blood cell types were identified; hyalinocytes, granulocytes, and serous cells. Haemocytes were classified based on the presence/absence of granules, and nucleus to cytoplasm ratio. Granulocytes were the most common cell type (62 ± 2.81%), followed by hyalinocytes (36 ± 2.35%), and serous cells (2 ± 0.90%). Granulocytes, and hyalinocytes were found to be immunologically active, with the ability to phagocytose Congo red stained yeast. Of the cells involved in phagocytosis, granulocytes were the most active with 88.8 ± 3.9% of these haemocytes engulfing yeast. Cytochemical stains (phenoloxidase, peroxidase, superoxide, melanin, neutral red) showed that enzymes associated with phagocytic activity were localised in granules within granulocytes. Based on their affinities for Giemsa/May-Grünwald stain, haemocytes were also defined as either acidic, basic or neutral. Hyalinocytes and serous cells were found to be eosinophilic, whilst granulocytes were either basophilic (large granulocytes), eosinophilic (small granulocytes) or a combination of the two (combination granulocytes). Light, differential interference contrast and epifluorescence microscopy identified three sub-populations of granulocytes based on size and granularity; small (4.00-5.00 lm in diameter, with small granules (0.05-0.5 lm in diameter), large (5.00-9.00 lm in diameter, with large granules (0.50-2.50 lm in diameter) and combination (5.00-9.00 lm in diameter, with both large and small granules). These observations demonstrate that P. imbricata have a variety of morphologically and functionally specialized haemocytes, many of which maybe associated with immunological functions.
Characterization of hemocytes from different body fluids of the eastern oyster Crassostrea virginica
Fish & shellfish immunology, 2017
Bivalve hemocytes are involved in a variety of physiological and immunological functions. Circulating hemocytes in the hemolymph represent the main component of the internal self-defense system while hemocytes present in the extrapallial space (between the mantle and the shell) are actively involved in biomineralization and shell formation. This study focused on the characterization of hemocytes from different body fluids of the eastern oyster Crassostrea virginica. Hemocytes present in the hemolymph were compared to those contained in the extrapallial fluid. Hemocytes associated with the mucus layer covering pallial organs (mantle, gills, body wall) were also investigated because of their potential role as sentinel cells. Hemocytes were characterized using flow cytometry in conjunction with fluorescent epitope markers (clusters of differentiation, lectins) as well as functional assays (i.e. phagocytosis and reactive oxygen species -ROS). Compared with the hemolymph, there was a sig...
2010
The morphology and cytochemistry of Pinctada imbricata haemocytes were studied in vitro. Three distinct blood cell types were identified; hyalinocytes, granulocytes, and serous cells. Haemocytes were classified based on the presence/absence of granules, and nucleus to cytoplasm ratio. Granulocytes were the most common cell type (62 ± 2.81%), followed by hyalinocytes (36 ± 2.35%), and serous cells (2 ± 0.90%). Granulocytes, and hyalinocytes were found to be immunologically active, with the ability to phagocytose Congo red stained yeast. Of the cells involved in phagocytosis, granulocytes were the most active with 88.8 ± 3.9% of these haemocytes engulfing yeast. Cytochemical stains (phenoloxidase, peroxidase, superoxide, melanin, neutral red) showed that enzymes associated with phagocytic activity were localised in granules within granulocytes. Based on their affinities for Giemsa/May-Grünwald stain, haemocytes were also defined as either acidic, basic or neutral. Hyalinocytes and serous cells were found to be eosinophilic, whilst granulocytes were either basophilic (large granulocytes), eosinophilic (small granulocytes) or a combination of the two (combination granulocytes). Light, differential interference contrast and epifluorescence microscopy identified three sub-populations of granulocytes based on size and granularity; small (4.00-5.00 lm in diameter, with small granules (0.05-0.5 lm in diameter), large (5.00-9.00 lm in diameter, with large granules (0.50-2.50 lm in diameter) and combination (5.00-9.00 lm in diameter, with both large and small granules). These observations demonstrate that P. imbricata have a variety of morphologically and functionally specialized haemocytes, many of which maybe associated with immunological functions.
Cytometric investigations on hemocytes of the American oyster, Crassostrea virginica
Tissue and Cell, 1988
Pericardial hemolymph was obtained from American Oysters (Crassosrrea virginica) and the hemocytes characterized by flow cytometry. The cells were found to have a broad unimodal size distribution with a median diameter of 7 micrometers. Total protein measured by flow cytometric fluorescence of dansylated cells also revealed a broad unimodal distribution similar to that obtained for size. The proportion of hemocytes in each stage of the cell cycle was measured using DNA-specific DAPI fluorescence. Histograms showed a single peak representing the Go/G, population. There was no evidence of S or G2+M phases of the cell cycle, nor was polyploidy seen. The forward and orthogonal light scatter of lixed hemocytes showed no evidence of sub-populations on the basis of cytoplasmic granularity. Thus, in terms of these parameters, oyster hemocytes appear to represent a single population exhibiting graded cellular differences.
First cytochemical study of haemocytes from the crab Carcinus aestuarii (Crustacea, Decapoda)
European Journal of Histochemistry, 2010
For the first time, a morphological study of haemocytes from the crab Carcinus aestuarii was carried out by means of light microscopy and differing cytochemical assays. Analysis of haemocyte size frequency distribution (performed by means of a Coulter Counter) revealed the presence of two distinct haemocyte fractions in C. aestuarii haemolymph, depending on cell size. The first fraction was of about 3-5 µm in diameter and 30-50 fL in volume, the second was of about 6-12 µm in diameter and over 200 fL in volume. Mean cell diameter and volume were 8.20±1.7 µm and 272.30±143.5 fL, respectively. Haemocytes observed under light microscope were distinguished in three cell types: granulocytes (28%; 11.94±1.43 µm in diameter) with evident cytoplasmic granules, semigranulocytes (27%; 12.38±1.76 µm in diameter) with less granules than granulocytes, and hyalinocytes (44%; 7.88±1.6 µm in diameter) without granules. In addition, a peculiar cell type was occasionally found (about 1%): it was 25-30 µm in diameter and had a great vacuole and a peripheral cytoplasm with granules. Granulocyte and semigranulocyte granules stained in vivo with Neutral Red, indicating that they were lysosomes. Giemsa's dye confirmed that granulocytes and semigranulocytes were larger than hyalinocytes. Pappenheim's panoptical staining and Ehrlich's triacid mixture allowed to distinguish granule-containing cells (including semigranulocytes) in acidophils (64%), basophils (35%) and neutrophils (1%). Hyalinocytes showed always a basophilic cytoplasm. Haemocytes were positive to the PAS reaction for carbohydrates, even if cytoplasm carbohydrate distribution varied among cell types. Lastly, lipids were found on cell membrane and in cytoplasm of all haemocyte types in the form of black spots produced after Sudan Black B staining. The morphological characterisation of C. aestuarii haemocytes by light microscopy was necessary before performing both ultrastructural and functional studies of circulating cells.
Journal of Experimental Marine Biology and Ecology, 2003
In this report, we provide detailed protocols for flow-cytometric characterization of haemocytes from the eastern oyster, Crassostrea virginica, and report effects of sudden temperature elevation upon haemocyte characteristics. Haemocytes were differentiated from other particles in the haemolymph using a combination of the DNA-binding fluorochrome SYBR Green and internal cell complexity. In formalin-fixed haemolymph, four distinct subpopulations of haemocytes were found: small hyalinocytes, large hyalinocytes, small granulocytes, and granulocytes. Hyalinocytes were numerically dominant, small and large together accounting for 60% of the haemocytes; granulocytes accounted for 30 -35%, and small granulocytes were least numerous at < 5%. These percentages were the same for haemolymph analysed immediately after it was withdrawn from living oysters (without fixation) as for haemolymph fixed with formalin, although small and large hyalinocytes were not always clearly differentiated into two distinct populations. There was a general trend for the fixed cells to appear in the flow cytometer to be larger and more complex than the fresh ones (except for granulocyte complexity).
Fish & Shellfish Immunology, 2008
This work presents the first detailed microscopic and functional analysis of the haemocytes of an abalone; the European Haliotis tuberculata. It is shown that in contrast to the situation in bivalves, only very few basophilic ''granulocytes'' could be found and exclusively with a histological stain. Neither flow cytometry, phase contrast observation nor transmission electron microscopy were able to detect any granular cells. The large majority of cells was constituted of ''hyalinocytes'', which could be sorted by flow cytometry, for the first time, into small (blast-like) and large cells. This permits a detailed analysis of haemocytes and especially of the lowly represented blast-like cells. The differences in haemolymph cell composition between bivalves and gastropods is reviewed in depth and discussed in view of the new data we present. Most of the abalone haemocytes analysed harbour many vacuoles, large glycogen deposits, lipid inclusions and acidic compartments. However, although the number of these ''inclusions'' was rather variable in between individual hyalinocytes, these experiments did not allow to discern subpopulations using these criteria, and the population appears more a v a i l a b l e a t w w w . s c i e n c e d i r e c t . c o m j o u r n a l h o m e p a g e : w w w . e l s e v i e r . c o m / l o c a t e / f s i Fish & Shellfish Immunology (2008) 24, 400e411
Helgoländer Meeresuntersuchungen, 1984
Hemocytes represent one of the most important defense mechanisms against foreign material in Mollusca. The morphology, hematological parameters and behaviour of hemolymph cells were studied in the southern quahog Mercenaria campechiensis, the eastern oyster Crassostrea virginica, and the blood ark Anadara ovalis challenged with the bacteria Vibrio vulnificus and V. anguillarum. Two general classes of hemocytes (granular and agranular) exist in C. virginica and M. campechiensis. In contrast, A. ovalis possesses 3 general classes (granular, agranular and erythrocytes). Three types of granules were identified by light microscopy. When hemolymph cells were studied by transmission electron microscopy, the cytoplasm of hemolymph cells was noted to contain many organelles, including electron dense granules. Both agranular and granular hemolymph cells were capable of colchicine-sensitive pseudopodial movement and spreading. The results indicate that marine bivalves possess hemolymph blood cells which may play a role in the internal defense paralleling mammalian phagocytes. The morphology of these cells, as determined by light, scanning and transmission electron microscopy, showed some similarity to mammalianmononuclear phagocytes. The sub-cellular events of molluscan hemocyte phagocytosis of V. vulnificus and V~ anguillarum were studied by both scanning and transmission electron microscopy. The role of these cells and the factors which govern their behavior are of economic and public health importance.