Single-cell algae plastic barrel closed expansion technology
2019-07-26 16:03:01
In the process of industrialized shellfish cultivation, a large amount of monilia algae needs to be cultivated as larvae's bait. The success of single cell algae cultivation is directly related to the success or failure of nursery. The traditional productive culture often uses small-scale glass bottles for first-class preservation, small tanks or glass jars for two-stage expansion, and large-pool three-level culture. Because its expansion is open, it is easily contaminated, so that the subsequent tertiary culture often fails. In order to solve this problem, nylon bags are currently used for closed-enlargement and the number of bags used for the expansion has reached several hundred. The authors have explored the expansion technology of plastic barrels for S. mongolicum in artificial breeding of shellfish and other shellfish. , is briefly introduced as follows:
1 culture device
A white drum-shaped plastic barrel with a commercial capacity of 50 L was used as a culture container (the greater the transparency, the better). Wash the bucket when it is used, then pour the boiled clean filtered seawater to more than 2/3 of the bucket; seal and seal it, ie, the bucket is sterilized at the same time with seawater. After the seawater in the bucket is completely cooled, open the cover and add nutrients according to the proportion. , Access to algae species, then connect the inflatable tube, then, the barrel mouth covered with double gauze, and then cover the nylon film.
2 medium
According to the different types of cultured algae, different culture fluid formulations are selected. The authors used the MAV culture solution to cultivate golden algae and Platymonas sp. The formula is as follows: 100 grams of KNO3, 2.5 grams of FeSO4.7H2O, 10 grams of KH2PO4, 0.25 grams of MnSO4.H2O, 20 grams of EDTA-Na2, and 1000 L of sterile seawater.
3 ways to expand
3. l Bottle-in-a-barrel expansion type A high-quality purebred species grown exponentially in a first grade glass bottle is used as an alga species for seed expansion. Inoculation density: gold algae, Chaetoceros sp. 50X104 cells/ml or more; Platymonas viciae 10x104 cells/ml or more . The first 1/3 barrel, with the growth of single cell algae, gradually increase the culture solution.
3.2 After the barrels are connected to the barrels and the seedlings are connected to the barrels, the unicellular algae in the barrels reach a certain density (Goldwort, Chaetoceros sp. 250X104 cells/ml; Platymonas density is 50x104 cells/ml), and the barrels can be further expanded. , that is, remove 2/3 of the algal fluid from the barrel and connect it to other barrels, leaving 1/3 of the algal fluid, and add the culture solution to continue cultivation. In this way, the barrel expansion tank can be expanded 2-3 times in a row, and generally the third barrel is no longer divided into buckets after the barrel is connected, so as to avoid the possible contamination due to the high number of times of the barrel separation.
4 matters needing attention
4. l Expand the seed with the culture system clean filtered seawater boiled, cooled, add nutrients prepared.
4.2 Before expanding the seed, hands should be washed, rubbed with alcohol to wipe the bacteria, and the amount of algae and the addition of culture liquid and the amount of salt added to the container should be reduced or measured, and the gauze and nylon membrane covering the mouth of the container should be used. Inflatable tubes must be boiled and used after sterilization.
4.3 barrels should be arranged in a well-lighted culture room, if in the hot season, it is best to arrange the barrels under a special ventilated four-sided ventilation, in order to avoid excessive temperature affect the growth of monocytic algae.
4.4 Inflate during the day and stop at night; inflate at noon during the high temperature season.
The 4.5 barrels shall be numbered and marked with the date of seed expansion so that the tertiary culture tanks can be inoculated in the order of seed expansion.
The 4.6 bottle-in-bucket, barrel-in-barrel, and barrel-pool are continuous inoculation processes. Algae must be pure and exponentially growing. Algae that have been contaminated and aged cannot be used for inoculation.
5 training effect
Practice has proved that using the above methods to expand and cultivate unicellular algae has the characteristics of fast growth, convenient operation and minimal pollution. The medium nursery with a size of 1000m3 water can be expanded with 150-200 barrels, and it can be used for subsequent tertiary pool cultivation. Provide good quality algae species.
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1 culture device
A white drum-shaped plastic barrel with a commercial capacity of 50 L was used as a culture container (the greater the transparency, the better). Wash the bucket when it is used, then pour the boiled clean filtered seawater to more than 2/3 of the bucket; seal and seal it, ie, the bucket is sterilized at the same time with seawater. After the seawater in the bucket is completely cooled, open the cover and add nutrients according to the proportion. , Access to algae species, then connect the inflatable tube, then, the barrel mouth covered with double gauze, and then cover the nylon film.
2 medium
According to the different types of cultured algae, different culture fluid formulations are selected. The authors used the MAV culture solution to cultivate golden algae and Platymonas sp. The formula is as follows: 100 grams of KNO3, 2.5 grams of FeSO4.7H2O, 10 grams of KH2PO4, 0.25 grams of MnSO4.H2O, 20 grams of EDTA-Na2, and 1000 L of sterile seawater.
3 ways to expand
3. l Bottle-in-a-barrel expansion type A high-quality purebred species grown exponentially in a first grade glass bottle is used as an alga species for seed expansion. Inoculation density: gold algae, Chaetoceros sp. 50X104 cells/ml or more; Platymonas viciae 10x104 cells/ml or more . The first 1/3 barrel, with the growth of single cell algae, gradually increase the culture solution.
3.2 After the barrels are connected to the barrels and the seedlings are connected to the barrels, the unicellular algae in the barrels reach a certain density (Goldwort, Chaetoceros sp. 250X104 cells/ml; Platymonas density is 50x104 cells/ml), and the barrels can be further expanded. , that is, remove 2/3 of the algal fluid from the barrel and connect it to other barrels, leaving 1/3 of the algal fluid, and add the culture solution to continue cultivation. In this way, the barrel expansion tank can be expanded 2-3 times in a row, and generally the third barrel is no longer divided into buckets after the barrel is connected, so as to avoid the possible contamination due to the high number of times of the barrel separation.
4 matters needing attention
4. l Expand the seed with the culture system clean filtered seawater boiled, cooled, add nutrients prepared.
4.2 Before expanding the seed, hands should be washed, rubbed with alcohol to wipe the bacteria, and the amount of algae and the addition of culture liquid and the amount of salt added to the container should be reduced or measured, and the gauze and nylon membrane covering the mouth of the container should be used. Inflatable tubes must be boiled and used after sterilization.
4.3 barrels should be arranged in a well-lighted culture room, if in the hot season, it is best to arrange the barrels under a special ventilated four-sided ventilation, in order to avoid excessive temperature affect the growth of monocytic algae.
4.4 Inflate during the day and stop at night; inflate at noon during the high temperature season.
The 4.5 barrels shall be numbered and marked with the date of seed expansion so that the tertiary culture tanks can be inoculated in the order of seed expansion.
The 4.6 bottle-in-bucket, barrel-in-barrel, and barrel-pool are continuous inoculation processes. Algae must be pure and exponentially growing. Algae that have been contaminated and aged cannot be used for inoculation.
5 training effect
Practice has proved that using the above methods to expand and cultivate unicellular algae has the characteristics of fast growth, convenient operation and minimal pollution. The medium nursery with a size of 1000m3 water can be expanded with 150-200 barrels, and it can be used for subsequent tertiary pool cultivation. Provide good quality algae species.
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