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Translation 2723 ffiICH1VES FISHERIES RESEARCH BOARD OF CANADA Translation Series No. .2723 • MorpholOgical and. physiOlOgiCal: study of the . of natural and : .acquired iMmunity in sOMe ftsh bloOd; from Indo-Chinese waters Durand Original. title: Etude• morphologigue et physiologique du sang, de 1' inununite naturelle et acquise chez quelques poissons Indo-Chinois From.: Annales de l'Institut. Oceanographique (Annals .of the Océanographic. Institute) . , . 25 : 109-206; 1950 ' Translated by the Translation Section Department of the Environment Department of the Environment Fisheries Research Board of Canada Ha Iifàxltabbratory -W.- S.- 1973 it .128 pages typescript F? i3 t2 r.;.)- ANNALES DE L'INSTITUT OCEANOGRAPHIQUE (ANNALS OF THE OCEANOGRAPHIC INSTITUTE) nv eLd Volume XXV, pagr:3 109 to 206. December 26, 1950. WORK CONDUCTED AT THE OCEANOGRAPHIC INSTITUTE OF INDOCHINA MORPHOLOGICAL AND PHYSIOLOGICAL STUDY OF THE BLOOD, OF NATURAL AND Arcerrprm TrimuNTry IN SOME FISH FROM INDOCHINESE WATERS by M. J. DURAND, Oceanographic Institute of Indochina j3 c2 .7;') SUMMARY PART ONE COMPARATIVE MORPHOLOGY OF THE BLOOD OF SOME MARINE AND FRESHWATER FISH OF INDOCHINA Bibliographical Survey Chapter One. - THE BLOOD OF MARINE FISH I.- STUDY MATERIAL p.1- 7 IL COMPOSITION OF NORMAL BLOOD p.2- 7 Diploprion bifasciatum p.2- 7 Plectorhynchus lineatus p.9 Epinephelus maculatus p.10 Acanthurus str4 g^,-; - p.11 Pseudoscarus nuchipunctatus p.13 Pomacanthus semicirculatus p.14 Platax orbicularis p.14 Amphiprion polylepis p.15 • Balistes stellatus p.16 Scolopsis dubiosus p.17 Pseudorhombus neglectus p.18 Seriola nigrofasciata p.19 Lutj anus sanguineus p.20 Lutjanus chrysotaenia p.20 Cynoglossus bilineatus p.21 Arius tha1assinus p.21 III.- DIFFERENTIAL WHITE CELL COUNT OF NORMAL BLOOD p.24 IV. SUMMARY AND CONCLUSIONS, CHAPTER I p.25 Chapter Two - THE BLOOD OF FRESHWATER FISH p.28 I. STUDY MATERIAL p.28 II. COMPOSITION OF NORMAL BLOOD Notopterus notopterus p.29 Catlacarpio siamensis p.30 Osteochilus me1anopleura p.30 Cyclochellichthys enoploides p.31 Puntiop1ites proctozisron p.32 Labeo (Morulius) chrysophekadion p.32 Wal1ago attu p.33 Belodontichthys dinema p.34 Cryptopterus apogon p.34 Pangasius sutchi p.35 Pangasianodon gigas p.36 Macrones nemurus p.37 Ophiocephalus micropeltes p.37 Pseudosciaena soldado p. 38 Oxyeleotris marmorata p.39 Anabas testudineus p.39 -2- 111. - DIFFERENTIAL WHITE CELL COUNT OF NORMAL BLOOD p.41 1V. - SUMMARY AND CONCLUSIONS„CHAPTER 11 p.42 PART TWO STUDY OF CELLULAR REACTIONS, OF IMAGOCYTOSIS AND NATURAL IMMUNITY IN SOME FISH FROM INDOCHINESE WATERS. p.44 STUDY MATERIAL AND RESEARCH METHODOLOGY P.44 CHAPTER 111 - EFFECT OF VARIOUS FACTORS ON HAEMATIC WHITE CELL COMPOSITION p.48 REMARKS ON LOCAL EOSINOPHILIA 1. - PHYSICAL AND PHYSIOLOGICAL FACTORS p.48 Temperature - Seasonal Rhythm - Inanition - Splenectomy p.48 1. - SOME CHEMICAL FACTORS p.53 M 1. Effect of Arsenious Acid on Anabas testudineus, p.53 2. Effect of Pilocarpine and Adrenalin: on Diploprion bifasciatum p.55 151FEEUFF7FIEEhs 1ineatus p.56 111. - REMARKS ON LOCAL EOSINOPHILIA p.57 Effect of Pilocarpine on the white cell count of the body - cavity of Plectorhynchus lineatus p.57 1V. - SUMMARY AND CONCLUSIONS, CHAPTER 111 p.60 CHAPTER 1V - STUDY OF CELLULAR REACTIONS, PHAGOCYTOSIS • AND NATURAL IMMUNITY p.61 1. - BIBLIOGRAPHICAL SURVEY p.61 11. - PHAGOCYTOSIS OBSERVED IN THE BLOOD OF FISH p.63 111. - PHAGOCYTOSIS AND CELLULAR REACTION OBSERVED IN THE BODY- CAVITY p.64 A) Experiments on the Marine Fish Plectorhynchus lineatus. p.64 B) Experiments on the Freshwater Fish Anabas testudineus. , p.69 -3- 1. - Piscine Strain Inoculation of Pseudomonas scissa and Pseudomonaà flliépreâéêhà • ophiocephali P: 77à Inoculation with Eberthella bbb ib,e) Inoculation with Shigella - Anthracoïdes - AlcaligeUeâ = Vibrio - Escherichia - Proteus p:âi Inoculation with Escherichia isolated from Anabas 2. - Human Strain plâà • Inoculation with Mycobacterium tuberculosis b i&kbbt p:àà Inoculation with Staphylococcus aureus Inoculation with Vibrio comma 666666 biti 1'3151 • Inoculation with Bacillus prodigiosus &&& itb‘b ih96 1V. - SOME OBSERVATIONS ON ACQUIRED IMMUNITY IN FISH &&&&&&& 1. Cellular Reaction and Phagocytosis in Vaccillméd Piàh, f3,iôô 2. Duration and Specificity of Immunity pàôà V. SUMMARY AND CONCLUSIONS, CHAPTER 1V && 15,164 BIBLIOGRAPHY pilM I/ • PART ONE COMPARATIVE MORPHOLOGY OF THE BLOOD OF SOME MARINE AND FRESHWATER FISH OF INDOCHINA Bibliographical survey The microscopic study of both normal and pathological blood of fish has been done by numerous authors. In 1908, SABRAZtS and MURATET mention, among the works which,had appeared up to that time on the haematology of fish th6se of CUÉNOT (1889 and 1891), HIRSCH- FELD (1897), GIGLIOTOS (1896), GRUNBERG and RAWITZ (1900). They note that RAWITZ's research was concerned with Scyllium catulus (a squaloid selachian), a ganoid (Acipenser ruthoenus L.) and Teleostei (Scorpaena porcus L.; Serranus scriba L.; Sargus vulgaris Geof.; Crenilabrus pavo C.V.). They indicate that one of RAWITZ's important results was that the blood of various fish is far from homogeneous. Crenilabrus pavo, for example, has two kinds of red corpuscles without transitional phases; Scyllium catulus and Sargus vulgaris display erythrocytolytic patterns without the appear- ance of blood platelets; these patterns are not found in other species; leukocytes in various degenerative stages are found in the circulating blood of Scyllium and Crenilabrus. Scorpaena, Sargus and Crenilabrus variation in fine Li have large leukocytes which exhibit considerable structure; the leukocytes of Scyllium and Acipenser are characterized by polymorphous granulation. According to RAW.ITZ, eosinophils are totally absent in Teleostei. In 1894, SABRAZES reported the absence of eosinophils in the blood of the sea-horse as well. Mbreover, in his work co-authored with MURATET, SABRAZES stated that GRUNBERG described the following cell forms observed in the blood of Scyllium catulus from the Berlin Aquarium: large and small mono- nuclear leukocytes occur in various shapes, depending on the size ratio of the nucleus (always large) to the cell body; the nucleus consists of a sharply defined chromatic structure; its shape is usually regular without marked variations; the leukocytes are oval, round or fusiform. In some, the protoplasm is homogeneous and greyer than the nucleus; others have a reticulated protoplasm with colourless lacunae. GRUNBERG also noted mononuclear leukocytes with a smaller, reniform nucleus; the cell body, which in some cases contains vacuoles, is much more developed than in the preceding types and stains unevenly.A third • - 2- class of leukocyte has a more or less convoluted, lobed nucleus. Poly- nuclear leukocytes, probably derived from the latter, as well as vacuolized, degenerate leukocytes are also found. Finally, the author reports spherical acidophilic granules in a number of leuko- cytes of the first group, irregular acidophilic granules in the ele- ments of the second group, and rod-like, acidophilic granules in the similar leukocytes of the third group and in the polynuclear group. He did not find any leukocytes with basophilic granulation. SIAWCILLO (1895), studying the blood of the three piscine species: Gobio fluviatilis, Cobitis fossilis and Raya denticulata, was able to confirm the presence of typical eosinophils in the blood of the ray. He also observed cells'in which the protoplasm is filled with fine, acicular crystals, also stainable by eosin. In the blood of the gudgeon, the same author observed cells containing granules of irregular shape and size, generally polyhedral and of soft consistency. MESNIL, who p qie same year carried out research on the resist- ance of fish u£eenMetions, experimented on Gobio fluviatilis, Perca fluviatilis and Carassius auratus. In the lymph of the gudgeon, MESNIL noticed rare cells of special appearance, which he described el"e : "Th,.se 1 eliknnyte c A if f er frnm the n thera only in that the i r protoplasm, instead of assuming the usual bluish tint, is slightly but not uniformly stainable by eosin; one sees sometimes a kind of pink mosaic pattern in which the various sections are separated by blue lines, and sometimes coarse, irregular granules of variable shape and size. Thus they are not true eosinophils as defined by Ehrlich, but rather pseudo-eosinophils."These are likely the same cells as were observed by SIAWCILLO in the blood of the same fish. • MESNIL mentions further that granulated cells are absent in the blood of the perch, as well as in Carassius auratus and certain narine species which he examined from that point of view: Merlangus vulgaris, Pagellus centrodontus, Conger vulgaris, Scomber scombrus and Platessa vulgaris. MEINERTZ (1902) described eosinophils in certain freshwater fish, particularly in Leuciscus rutilus, in the perch and the tench. SABRAZÉS and MURATET (1908) studied the blood of a Lophobranch, the sea-horse, Syngnathus hippocampus (Linnaeus), taken from the Arcachon bassin. These authors report the existence of two kinds of white corpuscles in the blood of this fish: -3- 1. Lymphocytes, of variable size: some round, measuring from 3.22 to 7.83 microns in diameter; others less regularly rounded, measuring 5 to 8.7 by 7 to 10.5 microns. According to the authors, these lymphocytes cannot be differentiated from those of mammals. The large nucleus is less basophilic than the protoplasm; a nucleole can sometimes be seen. Among these lymphocytes, there
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