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《Oceanologia Et Limnologia Sinica》 1999-01
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STUDIES ON THE RICKETTSIA-LIKE ORGANISM DISEASES OF TROPICAL MARINE PEARL OYSTER─THE MORPHOLOGY, MORPHOGENESIS AND ULTRASTRUCTURE OF RLO INCLUSIONS, AN AGENT FOR PINCTADA MAXIMA

WU Xin-zhong; PAN Jin-pei(South China Sea Institute of Oceanology,the Chinese Academy of Sciences,Guanhzhou,510301)  
The morphology, morphogenesis and ultrastructure of Rickettsia-like organism (RLO)inclusion in 5-month old diseased or moribund pearl oysters, Pinctada maxima, collected from theXinying Bay, Hainan Is1and in November 1993 and November 1994 were described. For ahistological examination the mantle, gills, hepatopancreas (including parts of stomoch and inteshne),rectum, adductor muscle, foot muscle and keber's organ were dissected and fixed in Bouln'sfixahve and 4% neutral formalin fixahve. Sechons were swined with rouhne Ehrich's H.E procedureand special methods including Gram-Weigert's stalning and Lehl-Neelsen and Wade-Fite foracid-fast staning. For scanning electron microscopy (SEM) analysis fresh specimens were fixed in2% glutaraldehyde. The selected small pieces (1mm2) of manue hssues were shaved with adoubleedged rasor blade and washed with a sodium cacodylate buffer (three time, 3othen/hme). Driedtissues with the shaved surfaces exposed were sputtercoated with goldpalladium. The characterishcsof RLO inclusion distribuhons in hssues showed that all kjnds of epithelial, connective tissue andendothelial cells are the target cells for RLO infechon. The morphology and size of inclusions fromdifferent tissues are different. The inclusions are Gramnegative but vallable with developing inclusions,Acid-fast-posihve in Which bright red when stained with Wade-Flte's ston and reddish black whenstrined with Ziehl Neelsen's stain.Pregranular and granular stages of developing inclusions were notedunder light microscopic examination. The granular stage may be dishngnished from stage I to stageⅢ (as, to asⅢ). At the pregranular stage (pgi) the inclusions do not contrin granules. Gram stainwas Poor and host cell has sligtihy hypertrophied. From granular stage I to stage Ⅲ, inclusionscontrin developing granules which became more (tens up to 100) and larger (1.0- 4.0u asmeasured in gs Ⅲ inclusions). Gram stain varied from pale red or pale blue to purplish red with b1ueand host cell becoming much more hypertrophied, at last ruptured. Utrastwturaly, twostereo-structure granules, small granules (average 645.85 x 359.O9Um) and large granules (avempe2 090. 93 x 1 556. O8pm), in inclusionpositive host cells were observed under SEM in which somesmall granules were undergoing transverse binary fission (Hg. ltb) and budding division (Hg. I:c),and the large granule was undergoing ruptured with organism escaping (Fig. 1.d). Combined thepresent resu1ts obseried by light and SEM with previous transmission electron microscopy. it issuggested that RLO inclusions consist of two different granules surrounded by a membrane-bound, inwhich the fine granules may be free individual RLO and the large granules may be RLOmicroclonies involved in phagolysosomes.
【Fund】: 中国科学院重点课题!KS85-121
【CateGory Index】: S944.3
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