First cytochemical study of haemocytes from the crab Carcinus aestuarii (Crustacea, Decapoda) (original) (raw)
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First cytochemical study of haemocytes from the crab Carcinus aestuarii (Crustacea
2013
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
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.
Comparative Clinical Pathology, 2008
The caecilians (Amphibia, Gymnophiona) constitute one of the least known groups of terrestrial vertebrates because most species live underground in quite inaccessible environments. Siphonops annulatus is an exclusively fossorial species and is the most extensively distributed caecilian in South America. Little is known of this order concerning circulating granulocytes, including their morphological and cytochemical structure and ultrastructure. This paper is part of a project covering the study of granulocytes in representative species of the order Amphibia. Blood extensions were carried out and submitted to Leishman, Toluidine Blue, Periodic acid Schiff, Sirius Red and hydrogen o-toluidine peroxide methods. Part of the samples was prepared for conventional transmission electron microscopy. Among granular leukocytes, mature and immature neutrophils and eosinophils were identified, plus basophils. The most frequent granulocyte encountered in S. annulatus peripheral blood is the neutrophil. This is a cell with a hyper-segmented nucleus and with a very clear cytoplasm when compared to the eosinophil, which presents large cytoplasmic acidophilic granules. On the other hand, the basophils present basophilic and metachromatic granules. Glycogen was detected in the cytoplasm of the neutrophils and eosinophils, while basic protein rich in amino acids was observed in the eosinophil's granules. Myeloperoxidase activity was detected in the cytoplasm of the neutrophils and eosinophils. Neutrophils were ultrastructurally detected with three types of small granules: eosinophils with large and small spherical granules and basophils with large spherical granules with lamellate structures.
Scylla tranquebarica revealed the presence of prohaemocytes, granular haemocytes, cystocytes, plasmatocytes and podocytes. The diagnostic characteristics of these cells conformed to the descriptions given by earlier authors in decapod crustaceans and insects. The spherule cells, adipohemocytes and oenocytoids have been found to be absent. Anucleate cells have been identified by both light and transmission electron microscopic observations. The discernibility of such anulceated cells added a new dimension to the concept of hemocytes origin and differentiation. The existence of anucleate granulocytes is construed to be a tenable phenomenon and situation considering the stem cells property that they undergo both symmetric and asymmetric divisions to maintan cell population in organisms. The total hemocyte count and the differential count indicated the defence potential of the species.
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.
Haemocytes of the clam Tapes philippinarum (Adams & Reeve, 1850): morphofunctional characterisation
Fish & Shellfish Immunology, 2000
Tapes philippinarum is a bivalve mollusc of the Pacific Ocean, successfully imported for human consumption into the northern Adriatic Sea (Europe). For better knowledge of its considerable adaptive ability in comparison with similar autochthonous species, a morpho-functional characterisation of its haemocytes was carried out with the establishment of short-term cell cultures (60 min at 25 C). Various methods of cytochemical staining identified four cell types in the haemolymph: granulocytes (48•05% 1•43), hyalinocytes (32•18% 0•99), haemoblasts (18•97% 0•63) and serous cells (0•8% 0•19). The granulocytes, possessing cytoplasmic granules with di#ering dye a$nity, included basophils, neutrophils and acidophils. Such granules stained vitally with Neutral Red, and correspond to lysosomes. Hydrolytic and oxidative enzymes were mainly detectable after stimulation in the presence of yeast cells. Both granulocytes and hyalinocytes were positive for alkaline phosphatase, non-specific esterase, peroxidase, and cytochrome C oxidase, whereas only granulocytes were positive for-glucuronidase, acid esterase, and arylsulphatase. Both cell types were competent phagocytes towards yeast and plasma had an opsonising e#ect. Moreover, the respiratory burst accompanied phagocytosis with superoxide anion production, recognisable through cytoplasmic deposits of formazan after treatment with nitro blue tetrazolium. Haemoblasts were small undi#erentiated cells which, due to their morphology and positivity to the anti-CD34 antibody, show the typical features of stem cells. Serous cells, probably arising from Keber's gland and belonging to another di#erentiation pathway, contained non-sulphate acid mucopolysaccharides and play an important role in early defence mechanisms, taking part in the formation of clots.
Haemocytes of the clam Tapes philippinarum (Adams & Reeve, 1850): morphofunctional characterisation
Fish & Shellfish Immunology, 2000
Tapes philippinarum is a bivalve mollusc of the Pacific Ocean, successfully imported for human consumption into the northern Adriatic Sea (Europe). For better knowledge of its considerable adaptive ability in comparison with similar autochthonous species, a morpho-functional characterisation of its haemocytes was carried out with the establishment of short-term cell cultures (60 min at 25 C). Various methods of cytochemical staining identified four cell types in the haemolymph: granulocytes (48·05% 1·43), hyalinocytes (32·18% 0·99), haemoblasts (18·97% 0·63) and serous cells (0·8% 0·19). The granulocytes, possessing cytoplasmic granules with di#ering dye a$nity, included basophils, neutrophils and acidophils. Such granules stained vitally with Neutral Red, and correspond to lysosomes. Hydrolytic and oxidative enzymes were mainly detectable after stimulation in the presence of yeast cells. Both granulocytes and hyalinocytes were positive for alkaline phosphatase, non-specific esterase, peroxidase, and cytochrome C oxidase, whereas only granulocytes were positive for -glucuronidase, acid esterase, and arylsulphatase. Both cell types were competent phagocytes towards yeast and plasma had an opsonising e#ect. Moreover, the respiratory burst accompanied phagocytosis with superoxide anion production, recognisable through cytoplasmic deposits of formazan after treatment with nitro blue tetrazolium. Haemoblasts were small undi#erentiated cells which, due to their morphology and positivity to the anti-CD34 antibody, show the typical features of stem cells. Serous cells, probably arising from Keber's gland and belonging to another di#erentiation pathway, contained non-sulphate acid mucopolysaccharides and play an important role in early defence mechanisms, taking part in the formation of clots.
Cytometric, morphologic and enzymatic characterisation of haemocytes in Anodonta cygnea
Comparative Biochemistry and Physiology A-molecular & Integrative Physiology, 2002
The haemocytes in bivalve mussels are involved in many processes such as lesion repair, shell repair, elimination of small particles and toxic substances. In Anodonta cygnea there are two categories of haemolymph cells, the granulocytes and hyalinocytes. Two groups of cells were identified by flow cytometry and morphological studies: one with larger size and granularity representing 75%, and another group of cells (25%) which were approximately half the size. The cytochemical reactions showed peroxidase activity in the larger cells and a weak prophenoloxidase activity in the smaller cells. These characteristics suggest that the most common haemocytes are granulocytes and hyalinocytes are less common. Enzymatic studies showed clear activities of few enzymes in different compartments of the mantle. Both haemocytes presented significant variations for a-manosidase and b-glucurosidase activities depending on the acid or alkaline pH. Almost all were sensitive to the pH changes, mainly the b-galactosidase in the haemolymph plasma. On the contrary, the same enzymatic analysis in the extrapallial elements showed more stabilised activities. The simulation of acidic and alkaline condition with the observation of significant morphological and enzymatic activity changes, allow us to speculate some functional role, mainly in the haemolymph elements. The granulocytes may be speculated to have intense involvement in the digestion of small residues with the formation of calcareous stores while the hyalinocytes are more responsible for the elimination of soluble cytotoxic compounds. ᮊ
International Journal of Fisheries and Aquaculture, 2013
For the first time haematological studies on freshwater crab P. masoniana (Henderson) has been carried out in Jammu, India. The aim of studies were to determine various type of hemocytes, their total and differential count as well. Based on morphological studies of hemocytes under microscope three types of haemocytes were identified viz., Hyalinocytes (H), Semigranulocytes (SG) and Granulocytes (G). These cells were varied with respect to their proportion, Semigranulocytes being the heighest (54%) follow by Granulocytes (30%) while Hyalinocytes were least (16%) among three cells.
Fish & Shellfish Immunology, 2007
For the first time, morpho-functional characterisation of haemocytes from the cockle Cerastoderma glaucum was performed to identify circulating cell types and to study their involvement in immune responses. Haemocyte mean number was 5.5 (Â10 5) cells/ mL haemolymph. Two main haemocyte types were found in haemolymph: granulocytes (85%), about 10 mm in diameter and with evident cytoplasmic granules, and hyalinocytes (15%), 8 to 14 mm in diameter, with a few or no granules. Most of the cytoplasmic granules stained in vivo with Neutral Red, indicating that they were lysosomes. On the basis of haemocyte staining properties, granulocytes and hyalinocytes were further classified as basophils and acidophils. Acidophil hyalinocytes were the largest haemocyte type (about 14 mm in diameter) and had an eccentric nucleus and a large cytoplasmic vacuole. Both granulocytes and hyalinocytes (except acidophils) were able to phagocytise yeast cells, although the basal phagocytic index was very low (about 2%). It increased significantly (up to 26%) after pre-incubation of yeast in cell-free haemolymph, suggesting that haemolymph has opsonising properties. Haemocytes also produced superoxide anion. Moreover, both granulocytes and hyalinocytes (except acidophils) were positive for some important hydrolytic and oxidative enzymes. Lysozyme-like activity was recorded in both cell-free haemolymph and haemocyte lysate, although enzyme activity in cell lysate was significantly higher. Results indicate that haemocytes from C. glaucum are effective cells in immune responses.