Since the basement algae are aquatic plants, their growth and reproduction are closely related to the water temperature. Therefore, the inoculation time should be selected during the breeding season, usually in the spring and autumn. It is best to be in spring, because it is the peak of its reproduction from March to May each year. Because the climate conditions in different places are different, the water temperature should be taken as the basic parameter, and it is most appropriate to inoculate in the season when the water temperature is 16~25°C. When the water temperature is lower than 16°C, the algae cells do not breed easily, and when the temperature is higher than 25°C, the algal cells are likely to rot and die.
Inoculation method
1. Natural reproduction and inoculation method: Take about 30g of cleaned algae species, and treat turtle shells that have been treated together with algae species in water tanks and other containers, and add clean culture water to adjust the pH to 7.5-6.5. Between the culture containers and the place where the scattered light is strong, the algal organisms are naturally released and fixed on the tortoise shell. Generally, after about 30 days, the turtle will be covered with a thin layer of green. Fluff, indicating successful vaccination.
2. Artificial inoculation method: The algae are cut with scissors, and then smashed with a wooden stick, with gauze to wrap the dilapidated algae, and the green juice spore fluid is filtered. The spore fluid contains a large amount of spores. Add 0.5 mL of VB1 solution per kilogram of spore fluid and 121 mL of VB, add the culture solution containing the appropriate amount of nutrient salts, and store it in a culture container, and then put the treated turtle species. This method is generally 25 to 30 days, the turtle's body surface will find green hair, indicating that the vaccination has been successful.
3. Medium inoculation method:
(1) Preparation of the medium: Accurately weighed 15 g potatoes, 20 g starch and 5 g sugar. Mix uniformly, place in a beaker, add water to 500 mL, and boil for 20 minutes. Filter and take the filtrate and add 16 g of agar to heat and melt. After cooling, the medium for inoculating the vegetative algae was prepared.
(2) Preparation of spore fluid: Take 500g of algae species and put them in a pot of water. After about 10 days, the water is dark green, the culture water temperature is kept at 20～24°C, and the hemocytometer is placed under the microscope. The concentration of spores in the culture solution was calculated. The culture solution was filtered when the spore concentration was greater than 100 per ml, and then the filtered culture liquid was placed in a centrifuge to centrifuge for 5 minutes at 1,500 revolutions per minute to obtain a spore concentrate.
(3) Inoculation: The medium is heated and melted. After cooling slightly, the medium is rubbed with a brush and wiped on a dried and processed turtle, and the medium is thinly evened, and the medium on the back of the turtle is not completely cooled. Use a clean brush to remove the spore concentrate and evenly apply it to the turtle's backing medium. After cooling, place the turtle into a transparent glass container filled with clean water. Place it in a bright place with scattered light, or use 20 ~ 40W fluorescent lamp irradiation fluorescent lamp 1m away from the water surface.
Problems to be aware of when inoculating:
(1) The size of the inoculation container should be appropriate. Inoculation of 100g of the turtle should be carried out in glass jars. One container should contain one and the turtle should be immersed in the water 2 to 3cm. The container should be tightly wrapped with a thin net cloth to avoid The turtle climbed out of the bottle.
(2) Do not change water or feed during inoculation. Changing the water will change the inoculation density of spores and spores, and turtles will excrete feces after feeding, resulting in spoilage of spores.
(3) Keep quiet: The place where turtles are vaccinated should be kept as quiet as possible so that the turtles can remain in a state of volcanic status and contribute to the survival of the genus Basidiomycetes.
Inoculation method
1. Natural reproduction and inoculation method: Take about 30g of cleaned algae species, and treat turtle shells that have been treated together with algae species in water tanks and other containers, and add clean culture water to adjust the pH to 7.5-6.5. Between the culture containers and the place where the scattered light is strong, the algal organisms are naturally released and fixed on the tortoise shell. Generally, after about 30 days, the turtle will be covered with a thin layer of green. Fluff, indicating successful vaccination.
2. Artificial inoculation method: The algae are cut with scissors, and then smashed with a wooden stick, with gauze to wrap the dilapidated algae, and the green juice spore fluid is filtered. The spore fluid contains a large amount of spores. Add 0.5 mL of VB1 solution per kilogram of spore fluid and 121 mL of VB, add the culture solution containing the appropriate amount of nutrient salts, and store it in a culture container, and then put the treated turtle species. This method is generally 25 to 30 days, the turtle's body surface will find green hair, indicating that the vaccination has been successful.
3. Medium inoculation method:
(1) Preparation of the medium: Accurately weighed 15 g potatoes, 20 g starch and 5 g sugar. Mix uniformly, place in a beaker, add water to 500 mL, and boil for 20 minutes. Filter and take the filtrate and add 16 g of agar to heat and melt. After cooling, the medium for inoculating the vegetative algae was prepared.
(2) Preparation of spore fluid: Take 500g of algae species and put them in a pot of water. After about 10 days, the water is dark green, the culture water temperature is kept at 20～24°C, and the hemocytometer is placed under the microscope. The concentration of spores in the culture solution was calculated. The culture solution was filtered when the spore concentration was greater than 100 per ml, and then the filtered culture liquid was placed in a centrifuge to centrifuge for 5 minutes at 1,500 revolutions per minute to obtain a spore concentrate.
(3) Inoculation: The medium is heated and melted. After cooling slightly, the medium is rubbed with a brush and wiped on a dried and processed turtle, and the medium is thinly evened, and the medium on the back of the turtle is not completely cooled. Use a clean brush to remove the spore concentrate and evenly apply it to the turtle's backing medium. After cooling, place the turtle into a transparent glass container filled with clean water. Place it in a bright place with scattered light, or use 20 ~ 40W fluorescent lamp irradiation fluorescent lamp 1m away from the water surface.
Problems to be aware of when inoculating:
(1) The size of the inoculation container should be appropriate. Inoculation of 100g of the turtle should be carried out in glass jars. One container should contain one and the turtle should be immersed in the water 2 to 3cm. The container should be tightly wrapped with a thin net cloth to avoid The turtle climbed out of the bottle.
(2) Do not change water or feed during inoculation. Changing the water will change the inoculation density of spores and spores, and turtles will excrete feces after feeding, resulting in spoilage of spores.
(3) Keep quiet: The place where turtles are vaccinated should be kept as quiet as possible so that the turtles can remain in a state of volcanic status and contribute to the survival of the genus Basidiomycetes.
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