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Four types of granulocytes, neutrophil, basophil, eosinophil and pithy granulocyte, were observed in peripheral blood of bowfin Amia calva. The neutrophil had three types of granules, eosinophilic granule (αG； orange), chromophobic granule (βG) and basophilic granule (γG； light blue) in the cytoplasm. These granules were round or oval, but the maximum diameter was different each other (αG, ≤0.5 μm; βG, ≤1.0 μm； γG, ≤0.3 μm). Alkaline phosphatase and peroxidase (PO) were detected in the βG, however, central part of the granule was negative. Basophil had round or oval granules (≤1.1 μm) which show violet. The granules were periodic acid Schiff (PAS) and toluidine blue (TB) positive. Eosinophil had two types of granules. One was major which was large and showed eosinophilic, another was minor with small size and basophilic. Both granules were PO positive (the former was weakly and the latter strongly). The latter was also PAS and TB positive. Pithy granulocyte was occasionally found in the blood. The granulocyte had ovoid granules with eosinophilic rod-shaped core and chromophobic surrounding part of the core.

Our observations of naturally lysed neutrophils from red sea-bream Pagrus major revealed that the fundamental neutrophil granule is chromophobic. It can be classified into two subpopulations based on the presence or absence of eosinophilic core.

Morphological and cytochemical characteristics of neutrophil from African lungfish, Protopterus annectens, were examined by light microscopy. The neutrophils were round to oval (35-40μm in diameter) and the nucleus round to lobule-shaped. Only one type of granule, panchromatophilic granule (PG) was observed in the neutrophil. The PG was round to rod-shaped and stained panchromatophilic according to staining conditions. The PG was eosinophilic or methylenophilic（methylene blue positive) with May-Gr?nwald (MG). And further, the PG showed reddish purple with Giemsa stain. This color tone may be due to the metachromatic reaction by azure B in Giemsa (metaazurophilic). This granule was acid phosphatase, α-naphtyl acetate esterase, α-naphtyl butyrate esterase, naphthol AS-D chloroacetate esterase, PAS (α-amylase digestion resistance) and toluidine blue (metachromatic) positive.

Morphological and cytochemical characteristics of neutrophil in Japanese amberjack, Seriola quinqueradiata were examined by light microscopy. The neutrophils were round to oval (7.0-11.0μm in diameter) and the nucleus round to lobule-shaped. Granules of the neutrophil were classified into three types； acidophilic granule (αG), chromophobic granule (βG), chromophobic granule (βG) and basophilic granule (γG). The αG was rod-shaped (0.5-1.0μm in length, 0.2μm in width) and stained with May-Grünwald (MG) stain. This granule was not observed in the preparations stained with Giemsa stain. The MG-Giemsa (MGG) staining pattern of the granule was influenced by pH and concentation of diluent of the staining solution. The βG was round to oval (≦0.5μm in diameter) and unstained by Romanovsky type stain (MG, Giemsa and MGG). This granule was alkaline phosphatase, peroxidase and sudan black B positive. The γG was round to oval (≦0.3μm in diameter) and stained with Giemsa and MGG, but not with MG. This granule was α-naphtyl acetate esterase positive. The Yasumoto body (Y-body) was also found in the neutrophil and toluidine blue positive.

Morphological and cytochemical characteristics of neutrophil in red sea-bream, Pagrus major were examined by light microscopy. The neutrophils were round to oval (7.5-11.5μm in diameter) and the nucleus round to kidney-shaped. Granules of the neutrophil were classified into two types； acidophilic granule (αG) and chromophobic granule (βG). The αG was round to oval (0.4 μm in diameter) and stained with May-Grünwald (MG) and MG-Giemsa (MGG) stain. The Giemsa staining pattern of the granule was influenced by pH and concentration of diluent of the staining solution. Some enzymes, such as acid phosphatase, α-naphtyl acetate esterase and naphthol AS-D chloroacetate esterase were detected in the αG. The βG was round to oval (≦0.5μm in diameter) and unstained by Romanowsky type stain (MG, Giemsa and MGG). This granule was peroxidase and sudan black B positive. The Yasumoto body (Y-body) was also found in the neutrophil and toluidine blue positive.

Morphological and cytochemical characteristics of neutrophil in Japanese eel, Anguilla japonica were examined by light microscopy. The neutrophils were round to oval (9.0-14.5μm in diameter) and the nucleus round to lobule-shaped. Granules of the neutrophil were classified into three types； acidophilic granule (αG), chromophobic granule (βG) and basophilic granule (γG). The αG was round to oval (0.3μm in diameter) or rod-shaped (≦1.0 μm in length) and stained with May-Grünwald (MG) stain. This granule was not observed in the preparations stained with Giemsa stain. The βG was round to oval (≦0.6μm in diameter) and unstained by Romanowsky type stain (MG,Giemsa and MG-Giemsa (MGG)). This granule was peroxidase positive. The γG was round to oval (≦0.3μm in diameter) and stained with Giemsa stain. Some enzymes, such as acid phosphatase, α-naphtyl acetate esterase and α-naphtyl butyrate esterase were detected in the γG. The Yasumoto body (Y-body) was also found in the neutrophil and toluidine blue positive.

Morphological and cytochemical characteristics of neutrophil in Polypterus endlicheri were examined by light microscopy and the composition of neutrophil granule were inferred in this study. The neutrophils were round or oval(14 .0 ～ 23.0 μm in diameter) and the nucleus round to two lobule-shaped. Three types of granules； two types of acidophilic granules (α-1G and α-2G), and basophilic granule (γG) were observed in neutrophil. Chromophobic granule (βG) was not observed. The α-1G was round (0.3μm in diameter) or rod (0.8μm in length) in shape and observed in May-Grünwald (MG), Giemsa and MG-Giemsan (MGG) preparations. The α-1G was alkaline phosphatase, acid phosphatase, β-glucronidase, α-naphtyl acetate esterase, α-naphtyl butyrate esterase, naphthol AS-D chloroacetate esterase and sudan black B positive. On the preparation fixed with formalin vapor, the α-1G was stained with eosin. The α-2G was round (0.3μm in diameter) and observed in MG and MGG preparations using 1/15M phosphate buffer (pH7.0 and pH8.0). The α-2G was stained with eosin on the preparation fixed with formalin-methanol. The γG was round to oval (

Ｍorphological and cytochemical characteristics of neutrophil in hagfish, Eptatretus burgeri were examined by light microscopy and the composition of neutrophil granule were inferred in this study. The neutrophils were round or oval (15-20μm in diameter) and the nucleus round to two lobule-shaped. Only one types of granule ； basophilic granule (γG) was observed in the neutrophil. The γG was round or oval (≦0.5μm in diameter) and stained with not only MG using distilled water and 5 mM phosphate buffer, but also Giemsa and MGG. Alpha-naphthyl butyrate esterase was detected in the γG. Yasumoto body (Y-body) was also observed in neutrophil and stained with toluidine blue.

Morphological and cytochemical characteristics of hemocyte in the seed shrimp, Vargula hilgendorfii, were examined by light microscopy. Only a single type of hemocyte (granulocyte) was observed in the hemolymph. This hemocyte was large (18-40 μm in diameter) and irregular in shape. Two types of round granules, large granule (LG) and small granule (SG) were observed in a hemocyte. The LG was not stained with May-Grünwald nor with HE. The SG was stained with eosin, safranin, methylene blue, toluidine blue, PAS, sudan black B. Phenoloxidase activity (DOPA reaction) was detected in both SG and hyaloplasm. However, the LG was not stained with these stains. These morphological and tinctorial characteristics are compared with those of other crustacean hemocytes.

Morphological and cytochemical characteristics of neutrophil in asian arowana, Scleropages formosus were examined by light microscopy and the composition of neutrophil granule were inferred in this study. The neutrophils were round or oval (12.0～17.0μm in diameter) and the nucleus round to three lobule-shaped. Three types of granules； acidophilic (eosinophilic) granule (αG), chromophobic granule (βG) and basophilic granule (γG) were observed in neutriphil. The αG was rod to spindle in shape (1.0×0.3μm) and observed in May-Grünwald (MG) preparation using distilled water and low molar (5mM) buffer. The acidophilicity of αG disappeared by Giemsa stain after MG. The βG was round to oval (

Morphological and cytochemical characteristics of neutrophil in the northern pike, Exos lucius were examined by light microscopy and the composition of neutrophil granule were inferred in this study. The neutrophils were round or oval, and the nucleus round to three lobule-shaped. The neutrophil possessed the chromophobic granule (βG) and Y-body. The βG was acid phosphatase and peroxidase positive. The Y-body was toluidine blue positive.