Method for preparing and culturing culture medium for supravital culture of seawater scuticociliatida ciliates

A technology of shield ciliate and culture medium, which is applied in the field of aquatic animal disease research, can solve problems such as complex culture medium formula, and achieve the effects of simple and easy culture method, less equipment and longer culture time.

Inactive Publication Date: 2012-06-13
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

They mainly use artificial synthetic medium containing various components to continuously culture ciliates in vitro. The medium formula is complex, and the culture time of ciliates is only about 10 days.

Method used

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  • Method for preparing and culturing culture medium for supravital culture of seawater scuticociliatida ciliates
  • Method for preparing and culturing culture medium for supravital culture of seawater scuticociliatida ciliates
  • Method for preparing and culturing culture medium for supravital culture of seawater scuticociliatida ciliates

Examples

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Embodiment 1

[0022] In June 2011, a turbot cultured in a turbot breeding factory in Yantai, Shandong Province developed shield ciliate disease. The diseased turbot was packaged and oxygenated and brought back to the laboratory of Qingdao Yellow Sea Fisheries Research Institute for pathogen isolation and cultivation.

[0023] In the laboratory, the in vivo medium of scutellum ciliates was first prepared. Select 20 healthy turbot brought back from the breeding factory, cut about 1g of its brain tissue and about 1g of its back skinned muscle, mix the two into a large centrifuge tube, add 20ml of sterilized seawater and mix well with the tissue After homogenizing by the pulper for 10 minutes, add 70ml of sterilized seawater to fully shake and mix evenly. Transfer 90ml of the mixed mixture to a 200ml glass bottle with a sealed cap, seal it well and place it in a water bath at 65°C for 2 hours, during which it shakes gently every 15 minutes. After the water bath, the glass bottle was naturally ...

Embodiment 2

[0031] In August 2011, flounder flounder raised in a flounder breeding factory in Jiaonan City, Qingdao, Shandong Province also suffered from scutellum ciliate disease. The sick flounder was packed and oxygenated and brought back to the laboratory of Qingdao Yellow Sea Fisheries Research Institute for pathogen isolation and cultivation.

[0032] In the laboratory, the in vivo medium of scutellum ciliates was first prepared. Select 30 healthy flounder brought back from the breeding factory, cut about 1.5g of its brain tissue and about 0.5g of its back skinned muscle, mix the two into a large centrifuge tube, add 20ml of sterilized seawater for tissue After homogenizing with a homogenizer for 10 minutes, add 90ml of sterilized seawater to fully shake and mix evenly. Transfer the mixed 110ml mixture to a 200ml glass bottle with a sealed cap, seal it and place it in a water bath at 70°C for 3 hours, shaking it every 15 minutes during the water bath. After the water bath, the glas...

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Abstract

The invention relates to a method for preparing and culturing a culture medium for the supravital culture of seawater scuticociliatida ciliates, and relates to the field of the study of aquatic animal diseases. The method for preparing the in-vitro culture medium comprises the following steps of: mixing fresh brain tissue and muscular tissue of paralichthys olivaceus or turbot fish in a mass ratio of (1:1)-(3:1), and homogenizing to form muddy flesh; adding sterilized seawater into the muddy flesh in a mass / volume ratio of the muddy flesh to the seawater of (1:45)-(1:55), and performing waterbath at the temperature of between 65 and 70 DEG C for 2 hours to prepare the in-vitro culture medium, wherein the prepared culture medium is stored at the temperature of 4 DEG C for a long time. Themethod for preparing the culture medium is simple, and complex components are not needed to be added; and the seawater scuticociliatida ciliates are subjected to the supravital culture by utilizing the culture medium prepared by the method, and the scuticociliatida ciliates can survive for over 30 days, so that the foundation is established for the further study of the pathogenesis and preventionand control technology of the scuticociliatida ciliates.

Description

Technical field: [0001] The invention belongs to the field of research on diseases of aquatic animals, and in particular relates to a method for realizing long-term cultivation of seawater shield ciliates in vitro of aquatic animals and preparation of culture medium thereof. Background technique: [0002] Scutellum ciliates are recognized as the main pathogens in mariculture, and many scholars at home and abroad have reported their serious harm to mariculture (Azusa Umehara, et al.2003, ESterud et al.2000). In general, shield ciliates live freely and feed on tiny organic particles (bacteria, protozoa, microalgae, organic debris, etc.) suspended in water. However, in the case of high-density culture, scutellum ciliates can usually become parasitic, feed on cells or tissue debris of echinoderms, shellfish, crustaceans and fish, and rapidly grow and proliferate in host tissues , and then cause deep tissue damage, forming serious lesions, and eventually lead to organ failure an...

Claims

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Application Information

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IPC IPC(8): C12N1/10C12R1/90
Inventor 张正李彬王印庚荣小军王岚廖梅杰陈贵平于雯雯
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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