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The Cultivation of Bonamia ostreae in Vitro and the Preparation of Polyclonal Antibody Against B. ostreae

Author: WangFei
Tutor: YangLianRu
School: Inner Mongolia Agricultural University
Course: Preventive Veterinary Medicine
Keywords: Bonamia ostreae Culture in vitro purification Fluorescence hybridization insitu Polyclonal antibody
CLC: S944
Type: Master's thesis
Year: 2013
Downloads: 1
Quote: 0
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Over the last few years, the outbreak of the Bonamiosis in several European countries resulted in serious mortalities of flat oysters and destroyed the oyster industries in these countries. However, the study of B.ostreae is still in primary stage. It is important to develop a reliable method to culture B.ostreae in vitro, especially for the studies of molecular mechanism of pathogenicity, the interaction between the host and parasite, and the disease managament.This study consists of three parts:1. The oysters suspected to be infected with B. ostreae were screened by PCR and the B.ostreae was cultured in vitro with host homogenate. Finall B.ostreae were demonstrated successfully cultured and passaged for three generations in vitro by in situ hybridization.2. B.ostreae were purified by Percoll gradient centrifugation and the quantities of pathogens for each layer were determined through RTQ-PCR.The results showed that the amount of B. ostreae for each layer ranged from9.63%(the layer of60%~70%) to29.08%(the layer of30%) Besides, the survival rates of B.ostreae were determined too.3. The polyclonal antibodies against B. ostreae were produced by inoculating mice with purified pathogens. The specificity of these polyclonal antibodies against B. ostreae was determined by western blot.In summary, this study developed a simple, feasible culturing method for B. ostreae in vitro. Moreover, the cultured pathogens were successfully purified and used for producing polyclonal antibodies against B. ostreae.

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CLC: > Agricultural Sciences > Aquaculture, fisheries > Fisheries Protection > Shellfish diseases, predators,and its control
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