A kind of separation and purification method of iodine algae in Honghu lake

A technology for separation and purification of iodine ammonium, applied in the biological field, can solve the problems of complex operation, lack of technical specifications, high cost, etc., and achieve the effect of simple process, convenient operation and low cost

Active Publication Date: 2021-12-28
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The existing polar capsule extraction method is realized by microfluidic chip technology based on quadrupole electrode array, but this method is complicated to operate, high in cost, and lacks technical specifications

Method used

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  • A kind of separation and purification method of iodine algae in Honghu lake
  • A kind of separation and purification method of iodine algae in Honghu lake
  • A kind of separation and purification method of iodine algae in Honghu lake

Examples

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Embodiment 1

[0023] The present invention is a separation method based on gradient centrifugation, and its key elements are centrifugal medium (sucrose, Percoll, etc.) and gradient composition. In order to determine the optimal method for the separation and purification of Honghu iodine polar capsules, a series of explorations and optimizations were carried out on the key elements of the gradient centrifugation steps:

[0024] Scheme 1 used sucrose with low cost as the centrifugation medium, and carried out the first exploration of polar capsule separation. The steps are as follows:

[0025] A. Take 5 mL of Honghu iodine spore fragmentation solution and add it to the bottom of a 50 mL centrifuge tube;

[0026] B. Through the syringe, inject the sucrose solutions with mass percentages of 40%, 45%, 50%, 55%, and 60% into the centrifuge tube in sequence, each layer is 8mL;

[0027] C, 3500rpm speed, centrifugal 15min;

[0028] Results: After centrifugation, the ratio of polar capsules to sh...

Embodiment 2

[0043] The separation and purification method of Honghu iodine polar capsules includes tissue sample homogenization, spore purification, ultrasonic crushing, Percoll gradient centrifugation, polar capsule recovery, secondary purification, resuspension and cleaning of recovered liquid, and the specific steps are as follows:

[0044] A. Tissue sample homogenization: cut out the crucian crucian throat that was found to contain Honghu iodine cysts by microscopic examination, put it into a hand-held homogenizer, and then add 0.1M PBS buffer to grind thoroughly until there is no obvious tissue in the homogenizer. organization block;

[0045]B. Purification of spores: filter the homogenate with double layers of medical gauze, collect the filtrate; centrifuge at 2500rpm for 15min, wash twice with 0.1M PBS buffer, resuspend the spores to 8mL with PBS, and add to the bottom of a 50mL centrifuge tube ; The sucrose solutions with mass percentages of 30%, 40%, 50%, and 60% were injected in...

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Abstract

The invention discloses a method for separating and purifying polar capsules of Honghu iodophobia, which comprises tissue sample homogenization, spore purification, ultrasonic crushing, Percoll density gradient centrifugation, polar capsule recovery, secondary purification, and recovery solution resuspension and cleaning. This method mainly utilizes the difference in mass proportion between polar capsules and other impurities to realize the separation of polar capsules and impurities, and uses 50%, 70%, 90% Percoll separation liquid to carry out density gradient centrifugation on the spore crushing liquid, and the 50% and 70% Percoll solutions Complete polar capsules with a purity close to 100% can be obtained between the layers and in the middle of the 70% Percoll separation solution. The present invention provides a new way for the source of myxosporidium polar cysts, and the purified polar cysts will help to study the biological characteristics of Honghu iodophora, analyze the weak link of life history, clarify the pathogenic mechanism and establish early detection means etc.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a method for separating and purifying polar capsules of Honghu iodine hyacinthia. Background technique [0002] Myxozoans are common parasites in farmed fish, and some species can cause major diseases of farmed fish. Among them, Honghu iodine worm is the pathogen of heterogeneous gibel crucian carp's "throat sporosis". It will form bean curd residue-like cysts in the throat of crucian carp, deprive the host of nutrients, hinder the fish from eating and cause mechanical compression, eventually leading to the death of the fish. The mortality rate of "throat spore disease" can exceed 90%, which seriously endangers the development of heterogeneous gibel crucian carp aquaculture industry. At present, the prevention and control of "laryngosporia" is facing several bottlenecks. On the one hand, the biological characteristics and pathogenic mechanism of Honghu iodophora...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/10C12N1/02C12R1/90
CPCC12N1/10C12N1/02
Inventor 顾泽茂郭庆祥翟艳花柳阳
Owner HUAZHONG AGRI UNIV
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