CYTOCHEMICAL FEATURES OF SHRIMP HEMOCYTES (original) (raw)
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Selected morpho-chemical features of hemocytes in farmed shrimp, Fenneropenaeus indicus in Iran
The aim of the present study was to determine various types of hemocytes, total and differential hemocyte count and to show some indices of phagocytosis, including percent phagocytosis and phagocytic index in Indian white shrimp, Fenneropenaeus indicus in Iranian shrimp farms. The hemolymph was extracted from the shrimps and mixed with anticoagulant. It then stained and Yeast was added as foreign particles to samples. Based on the cell size and presence of the granules and nucleocytoplasmic ratio, three major groups of hemocytes, including hyaline cells, small granular cells (SGC) and large granular cells (LGC) were identified. Hyaline cell (HC) was the smallest hemocyte. HC (hyaline cell) about was 10-15%, lower quantity in comparison to some species and those of LGC and SGC were 20-25% and 60-65%, respectively, suggesting that relative percentage of differential hemocyte count in Fenneropenaeus indicus is different from other decapods. Also, in this study, it was shown that SGH and LGH could engulf the yeast particles. In Contrast to some previous studies, no engulfment was observed by hyaline cells in vitro. In this study it was showed that there are some variations in total and differential hemocyte count compare to other species in the family of Penaeidae.
Morphological Characterization of the Hemocytes of the Clam,Ruditapes decussatus(Mollusca: Bivalvia)
Journal of Invertebrate Pathology, 1997
Josef Berger and Klára Slavíčková (2008) Morphological characterization of hemocytes in the adult linden bug, Pyrrhocoris apterus (L.) (Heteroptera). Zoological Studies 47(4): xxx-xxx. The morphology of hemocytes of the adult linden bug, Pyrrhocoris apterus (L.) (Heteroptera) was characterized via light (both classical and confocal) and transmission electron microscopy. Based on this characterization, we classified hemocytes into 4 distinct types: prohemocytes, granulocytes, plasmatocytes, and spherulocytes. Differential hemocyte counts are described. We propose using the Pappenheim panoptic stain to identify different hemocyte populations of P. apterus. These results are the beginning building blocks of a knowledge base for our ongoing studies of the function of P. apterus hemocytes. As certain morphological characteristics are similar to mammalian blood cells which are used in hemotoxicological safety screening, the use of P. apterus as a new hematotoxicological biomodel is suggested.
International Journal of Cell Biology, 2016
Hemocytes represent one of the most important defense mechanisms against foreign material in Crustacea and are also involved in a variety of other physiological responses. Fluorescent lectin-binding assays and cytochemical reactions were used to identify specificity and distribution of carbohydrate moieties and presence of several hydrolytic enzymes, in hemocytes of whiteleg shrimpLitopenaeus vannamei. Two general classes of circulating hemocytes (granular and agranular) exist inL. vannamei, which express carbohydrates residues for FITC-conjugated lectins WGA, LEA, and PNA; UEA and Con-A were not observed. Enzymatic studies indicated that acid phosphatase, nonspecific esterase, and specific esterases were present; alkaline phosphatase was not observed. The enzymes and carbohydrates are useful tools in hemocyte classification and cellular defense mechanism studies.
Morphofunctional study of hemocytes from lions-paw scallop Nodipecten subnodosus
Immunobiology, 2013
Hemocytes play an important role in internal defense in mollusk bivalves; they are generally divided into granular and hyaline types, where the granules possess molecules to combat foreign particles. We investigated the morphology, staining reactions, and immune-related activities of hemocytes from the pectinid lions-paw scallop Nodipecten subnodosus. We showed that, in contrast to the conditions in most bivalves and similar to other pectinids, no granular cells were observed; only very few semi-granular cells were present and the large majority of cells were hyalinocytes and blast-like cells. The cytoplasm in many hyalinocytes showed vesicles of various sizes, but none with the staining characteristics of typical granules. When hemolymph was removed from these scallops, the plasma did not clot and the hemocytes rapidly adhered to one another in suspension. When living hemocytes were placed on a glass surface, they clustered into groups of various sizes; within minutes, pseudopods extended from the cells and outward migration began. Many cells spread to form extensive networks of flat cells, and other undifferentiated cells did not spread and kept a rounded morphology. Some hemocytes were immunologically active because they phagocytize Escherichia coli bacteria. Hemocytes expressed the carbohydrates residues N-acetyl-d-glucosamine, ␣-d-glucose, ␣-d-mannose, and ␣-L-fucose moieties, and cytochemical assays showed activity of acid phosphatase, specific esterase, and non-specific esterase, enzymes commonly associated with lysosomes and immunologic activity. These observations serve as a useful tool for further investigation of morphofunctional studies and physiological status of scallops.
Journal of Morphology, 1987
The architecture and fine structure of the epigastric hematopoietic nodules of the ridgeback prawn, Sicyonia ingentis, are described. The nodules consist of a highly branched series of tubules that contain the maturing hemocytes within a connective tissue stroma. Hemocytes can exit the hematopoietic nodules by penetrating through fenestrations in the endothelial cell layer into the central hemal space or by migrating through the outer later of capsular cells and associated collagen fibrils. Four hemocyte categories were observed: agranular, small granule with cytoplasmic deposits, small granule without cytoplasmic deposits, and large granule hemocytes. This classification was based upon the presence, size, and type of cytoplasmic granules and the presence of cytoplasmic deposits. Only agranular cells and small granule hemocytes without cytoplasmic deposits appeared capable of division. Intermediate stages were observed between agranular hemocytes and small granule hemocytes with deposits and between small granule hemocytes without deposits and large granule hemocytes, suggesting existence of two distinct hemocyte lines.
Pakistan Veterinary Journal, 2013
The Litopenaeus vannamei is a prime aquaculture species and has a worth market value all over the world. In this study, we investigated the genesis and morphology of hematopoietic tissue (HPT) and types of hemocyte based on its morphology and cytochemical feature using histological and histochemical methods. The results revealed that HPT in L. vannamei is not visible histologically in mysis and early two post larvae (P 1 & P 2), and emerges in P 3 which locates in epigastric region. The HPT showed a continuous propagation of cells in successive post larval stages. Four types of cells can be identified in the HPT from adult specimen and mitotic activity is visible in the HPT. The hemocytes have been differentiated into five types and some of these types are co-related with the HPT cells. The cytochemical studies suggested that type I and II cells of hemocytes are PAS positive while scanty presence of prophenoloxidase was observed in type I cells. Similarly the type I cells are Sudan Black B positive and rest of the cells showed weak activity against lipid detective stain. This is the first effort towards genesis of HPT and its relation with circulating hemocytes in L. vannamei.
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.
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.
Journal of Experimental Zoology, 1977
Localization of c l o t t a b l e protein (CP) i n hemocytes of the Japanese horseshoe crab, Tachypleus tridentatus, was investigated by a fluorescent antibody technique. minority of hemocytes (15%) were stained strongly and uniformly while those i n the majority of hemocytes (85%) were not. cytoplasmic matrix of the majority was stained with various degrees of intensity and t h a t of the minority was not. The two types of hemocytes which showed d i f f e r e n t i n t r a c e l l u l a r localization of CP might be i n d i f f e r e n t maturational stages o r m i g h t play d i f f e r e n t roles in blood coagulation.