CN107372370B - Hatching solution for hatching aedes albopictus eggs and hatching method for aedes albopictus eggs - Google Patents
Hatching solution for hatching aedes albopictus eggs and hatching method for aedes albopictus eggs Download PDFInfo
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- CN107372370B CN107372370B CN201710803952.0A CN201710803952A CN107372370B CN 107372370 B CN107372370 B CN 107372370B CN 201710803952 A CN201710803952 A CN 201710803952A CN 107372370 B CN107372370 B CN 107372370B
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- 230000012447 hatching Effects 0.000 title claims abstract description 174
- 235000013601 eggs Nutrition 0.000 title claims abstract description 94
- 241000256173 Aedes albopictus Species 0.000 title claims abstract description 72
- 238000000034 method Methods 0.000 title claims abstract description 28
- 238000011534 incubation Methods 0.000 claims abstract description 58
- 239000012530 fluid Substances 0.000 claims abstract description 46
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 43
- 239000001888 Peptone Substances 0.000 claims abstract description 42
- 108010080698 Peptones Proteins 0.000 claims abstract description 42
- 235000019319 peptone Nutrition 0.000 claims abstract description 42
- 239000000243 solution Substances 0.000 claims abstract description 38
- 239000007864 aqueous solution Substances 0.000 claims abstract description 35
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- 241000255925 Diptera Species 0.000 claims abstract description 19
- 238000007789 sealing Methods 0.000 claims abstract description 14
- 238000005303 weighing Methods 0.000 claims abstract description 13
- 239000007788 liquid Substances 0.000 claims abstract description 7
- 238000000855 fermentation Methods 0.000 claims description 6
- 230000004151 fermentation Effects 0.000 claims description 6
- 230000003068 static effect Effects 0.000 claims 2
- 230000004083 survival effect Effects 0.000 abstract description 11
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 35
- 230000000052 comparative effect Effects 0.000 description 13
- 241000238631 Hexapoda Species 0.000 description 7
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 7
- 239000001301 oxygen Substances 0.000 description 7
- 229910052760 oxygen Inorganic materials 0.000 description 7
- 230000000694 effects Effects 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 4
- 241000607479 Yersinia pestis Species 0.000 description 3
- 238000009395 breeding Methods 0.000 description 3
- 230000001488 breeding effect Effects 0.000 description 3
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000005265 energy consumption Methods 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 230000001418 larval effect Effects 0.000 description 2
- 208000001490 Dengue Diseases 0.000 description 1
- 206010012310 Dengue fever Diseases 0.000 description 1
- 206010014596 Encephalitis Japanese B Diseases 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 201000006353 Filariasis Diseases 0.000 description 1
- 108010058643 Fungal Proteins Proteins 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- 201000005807 Japanese encephalitis Diseases 0.000 description 1
- 241000710842 Japanese encephalitis virus Species 0.000 description 1
- 101001061492 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) Partitioning protein REP1 Proteins 0.000 description 1
- 208000003152 Yellow Fever Diseases 0.000 description 1
- 208000020329 Zika virus infectious disease Diseases 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
Abstract
The invention relates to an incubation liquid for incubating aedes albopictus eggs and an incubation method of aedes albopictus eggs. The hatching solution is an aqueous solution prepared from yeast peptone and water, and the concentration of the yeast peptone in the aqueous solution is 0.7-1.5 g/L. The hatching method comprises the following steps: fermenting the hatching solution for hatching the aedes albopictus eggs; weighing aedes albopictus eggs, adding the aedes albopictus eggs into an incubation tube, adding the fermented incubation liquid into the incubation tube, sealing a tube cover of the incubation tube, shaking uniformly, standing for incubation, then opening the tube cover of the incubation tube, and transferring the incubated larvae into a feeding tray. The hatching fluid is used for hatching the aedes albopictus eggs under certain conditions, so that the hatching rate of the aedes albopictus eggs can be greatly increased, the hatching rate of active mosquito eggs in 1 hour is high, and the larva growth synchronism is good. The hatching method has the advantages of high larva survival rate, simple operation and low hatching cost.
Description
Technical Field
The invention relates to the technical field of artificial breeding of aedes albopictus, in particular to an incubation liquid for incubating aedes albopictus eggs and an incubation method of aedes albopictus eggs.
Background
The male sterile insect release technology is used as a new technology in pest control, and by cultivating the male sterile insects of pests and releasing the male sterile insects to a pest breeding place, the male sterile insects can not propagate offspring after mating with wild female insects, so that the insects can be reduced and even eliminated. Compared with the traditional pesticide disinsection strategy, the technology is environment-friendly and has no influence on other insects and the environment. Mosquitoes serve as intermediate hosts for other pathogens such as dengue fever, Zika, malaria, yellow fever, filariasis, Japanese encephalitis, and the like. In a large-scale male Aedes albopictus release plan, the incubation of the mosquito eggs is an important link, so that a simple, efficient and stable mosquito egg incubation method is needed.
The hatching rate of the aedes albopictus eggs in an anaerobic environment is much faster than that in an aerobic environment, and the traditional hatching mode of the aedes albopictus eggs is to remove oxygen by heating water to a saturation temperature under normal pressure by thermal power, so that the solubility of oxygen in water is reduced, and the oxygen escapes from the water to form the anaerobic environment. Then quickly transferring the oxygen-free water into a closed container, filling and sealing the container, cooling to 25-30 ℃, weighing the required mosquito eggs, opening the cover, pouring in the eggs, shaking uniformly, sealing the cover again, incubating at the temperature of 25-30 ℃ for 1 hour, finally opening the container, and transferring the incubated larvae into a feeding tray. The traditional technology has a general effect of promoting the hatching of Aedes albopictus, less than 80% of active mosquito eggs can be hatched within 1 hour, the rest mosquito eggs are gradually hatched after being transferred to a feeding tray, and the larva growth synchronism is poor. The oxygen content in the closed container is low, and the larvae hatched out first easily die. The incubation process needs heating, the energy consumption is high, the operation is complicated, and the requirement on the tightness of the container is strict. Is not beneficial to the standardized operation and cost control of mass production.
Disclosure of Invention
Based on the hatching solution, the hatching solution for hatching the aedes albopictus eggs can greatly accelerate the hatching rate of the aedes albopictus eggs, the hatching rate of active mosquito eggs in 1 hour is high, and the growth synchronism of larvae is good.
The specific technical scheme is as follows:
a hatching solution for hatching the eggs of Aedes albopictus is an aqueous solution prepared from yeast peptone and water, wherein the concentration of the yeast peptone in the aqueous solution is 0.7-1.5 g/L.
Preferably, the concentration of yeast peptone in the aqueous solution is 0.8-1.0 g/L.
Preferably, the concentration of yeast peptone in the aqueous solution is 0.9 g/L.
Preferably, the yeast peptone is Angel yeast peptone.
The invention also provides a hatching method of the aedes albopictus mosquito eggs. The method can greatly accelerate the hatching rate of the mosquito eggs of Aedes albopictus, the hatching rate of the active mosquito eggs in 1 hour is high, and the growth synchronism of the larvae is good.
The specific technical scheme is as follows:
an incubation method of aedes albopictus eggs comprises the following steps:
fermenting the hatching solution for hatching the aedes albopictus eggs to obtain the fermented hatching solution; the hatching solution is an aqueous solution prepared from yeast peptone and water, and the concentration of the yeast peptone in the aqueous solution is 0.7-1.5 g/L;
weighing aedes albopictus eggs, adding the aedes albopictus eggs into an incubation tube, adding the fermented incubation liquid into the incubation tube, sealing a tube cover of the incubation tube, shaking uniformly, standing for incubation, opening the tube cover of the incubation tube after incubation is finished, and transferring the incubated larvae into a feeding tray.
Preferably, the hatching solution is an aqueous solution prepared from yeast peptone and water, and the concentration of the yeast peptone in the aqueous solution is 0.8-1.0 g/L.
Preferably, the hatching solution is an aqueous solution prepared from yeast peptone and water, and the concentration of the yeast peptone in the aqueous solution is 0.9 g/L.
Preferably, the yeast peptone is Angel yeast peptone.
Preferably, the temperature of the fermentation is 25-30 ℃, and the time of the fermentation is 20-28 hours.
Preferably, the addition amount of the hatching fluid after fermentation is 50-80% of the volume of the hatching tube.
Preferably, the addition amount of the hatching fluid after fermentation is 60-70% of the volume of the hatching tube.
Preferably, the ratio of the aedes albopictus eggs to the fermented hatching fluid is 1 g: 150 and 350 mL.
Preferably, the temperature of the standing incubation is 25-30 ℃, and the time of the standing incubation is 0.8-1.2 hours.
The hatching solution for hatching the aedes albopictus eggs and the hatching method of the aedes albopictus eggs have the following advantages and beneficial effects:
the inventor of the invention unexpectedly discovers that a stable and efficient hatching method of aedes albopictus eggs can be obtained by using yeast peptone to prepare an aqueous solution with a specific concentration as a hatching solution for hatching the aedes albopictus eggs in long-term experience accumulation and experiment processes, the method can greatly accelerate the hatching rate of the aedes albopictus eggs, and the hatching effect is far better than that of the existing hatching method for removing oxygen by heating. The hatching solution of the aedes albopictus eggs is prepared by using yeast peptone, the hatching solution is fermented firstly when in use, and the yeast colonies which are propagated and increased in quantity consume oxygen in the solution through the respiration effect during the fermentation, so that an anaerobic environment which can promote the rapid hatching of the aedes albopictus eggs is created. The hatching fluid is used for hatching the aedes albopictus eggs under certain conditions, so that the hatching rate of the aedes albopictus eggs can be greatly increased, the hatching rate of active mosquito eggs in 1 hour is high, and the larva growth synchronism is good.
The hatching method of the invention has no strict requirement on the tightness of the hatching tube, and the hatching tube is more favorable for hatching effect if a certain amount of oxygen is left, so that hatched larvae can not die due to oxygen deficiency in a short time, the survival rate of the larvae is high, and the method creates a prerequisite for further large-scale standardized breeding of aedes albopictus.
According to the hatching method, only yeast peptone and water are needed for preparing the hatching solution, the preparation process and the operation process of hatching are very simple, energy consumption is not needed, and the hatching cost is greatly reduced.
Detailed Description
The incubation liquid for incubating aedes albopictus eggs and the incubation method of aedes albopictus eggs according to the present invention will be described in further detail with reference to specific embodiments.
The yeast peptone used in the following examples is Angel yeast peptone FP101, which is a stable, safe and comprehensive nutrient peptone prepared from pure cultured high protein baker's yeast by separating and enriching yeast protein and complex enzyme action. The contents thereof are as follows, total nitrogen: not less than 10.0%, a-amino nitrogen: not less than 2.5%, water content: less than or equal to 6.0 percent, NaCl: 2.0% or less, ash content: less than or equal to 15.0%, pH (2% solution): 5.3-7.2.
example 1
The embodiment provides a hatching method of aedes albopictus eggs, which comprises the following steps:
adding water into yeast peptone to prepare an aqueous solution with the concentration of 0.9g/L, and taking the aqueous solution as hatching fluid;
placing the hatching solution at the temperature of 25-30 ℃ for fermenting for 24 hours to obtain fermented hatching solution;
weighing 0.14g of aedes albopictus eggs, adding the eggs into a 50mL hatching tube, and adding 33.3mL of fermented hatching fluid (the fermented hatching fluid accounts for 2/3 of the volume of the hatching tube) into the hatching tube; sealing the tube cover of the incubation tube, shaking uniformly, horizontally placing, standing at 25-30 ℃ for incubation for 1 hour, then opening the tube cover of the incubation tube, and transferring the incubated larvae to a feeding tray.
Example 2
The embodiment provides a hatching method of aedes albopictus eggs, which comprises the following steps:
adding water into yeast peptone to prepare an aqueous solution with the concentration of 0.9g/L, and taking the aqueous solution as hatching fluid;
placing the hatching solution at the temperature of 25-30 ℃ for fermenting for 24 hours to obtain fermented hatching solution;
weighing 0.14g of aedes albopictus eggs, adding the eggs into a 50mL hatching tube, and adding 25mL of fermented hatching fluid into the hatching tube (the fermented hatching fluid accounts for 50% of the volume of the hatching tube); sealing the tube cover of the incubation tube, shaking up, standing and incubating for 1 hour at the temperature of 25-30 ℃, then opening the tube cover of the incubation tube, and transferring the incubated larvae to a feeding tray.
Example 3
The embodiment provides a hatching method of aedes albopictus eggs, which comprises the following steps:
adding water into yeast peptone to prepare an aqueous solution with the concentration of 0.9g/L, and taking the aqueous solution as hatching fluid;
placing the hatching solution at the temperature of 25-30 ℃ for fermenting for 24 hours to obtain fermented hatching solution;
weighing 0.14g of aedes albopictus eggs, adding the eggs into a 50mL hatching tube, and adding 40mL of fermented hatching fluid into the hatching tube (the fermented hatching fluid accounts for 80% of the volume of the hatching tube); sealing the tube cover of the incubation tube, shaking up, standing and incubating for 1 hour at the temperature of 25-30 ℃, then opening the tube cover of the incubation tube, and transferring the incubated larvae to a feeding tray.
Example 4
The embodiment provides a hatching method of aedes albopictus eggs, which comprises the following steps:
adding water into yeast peptone to prepare an aqueous solution with the concentration of 0.8g/L, and taking the aqueous solution as hatching fluid;
placing the hatching solution at the temperature of 25-30 ℃ for fermenting for 24 hours to obtain fermented hatching solution;
weighing 0.14g of aedes albopictus eggs, adding the eggs into a 50mL hatching tube, and adding 33.3mL of fermented hatching fluid (the fermented hatching fluid accounts for 2/3 of the volume of the hatching tube) into the hatching tube; sealing the tube cover of the incubation tube, shaking up, standing and incubating for 1 hour at the temperature of 25-30 ℃, then opening the tube cover of the incubation tube, and transferring the incubated larvae to a feeding tray.
Example 5
The embodiment provides a hatching method of aedes albopictus eggs, which comprises the following steps:
adding water into yeast peptone to prepare an aqueous solution with the concentration of 1.4g/L, and taking the aqueous solution as hatching fluid;
placing the hatching solution at the temperature of 25-30 ℃ for fermenting for 24 hours to obtain fermented hatching solution;
weighing 0.14g of aedes albopictus eggs, adding the eggs into a 50mL hatching tube, and adding 33.3mL of fermented hatching fluid (the fermented hatching fluid accounts for 2/3 of the volume of the hatching tube) into the hatching tube; sealing the tube cover of the incubation tube, shaking up, standing and incubating for 1 hour at the temperature of 25-30 ℃, then opening the tube cover of the incubation tube, and transferring the incubated larvae to a feeding tray.
Comparative example 1
The embodiment provides a hatching method of aedes albopictus eggs, which comprises the following steps:
adding water into yeast peptone to prepare an aqueous solution with the concentration of 2.0g/L, and taking the aqueous solution as hatching fluid;
placing the hatching solution at the temperature of 25-30 ℃ for fermenting for 24 hours to obtain fermented hatching solution;
weighing 0.14g of aedes albopictus eggs, adding the eggs into a 50mL hatching tube, and adding 33.3mL of fermented hatching fluid (the fermented hatching fluid accounts for 2/3 of the volume of the hatching tube) into the hatching tube; sealing the tube cover of the incubation tube, shaking up, standing and incubating for 1 hour at the temperature of 25-30 ℃, then opening the tube cover of the incubation tube, and transferring the incubated larvae to a feeding tray.
Comparative example 2
The embodiment provides a hatching method of aedes albopictus eggs, which comprises the following steps:
adding water into yeast peptone to prepare an aqueous solution with the concentration of 0.1g/L, and taking the aqueous solution as hatching fluid;
placing the hatching solution at the temperature of 25-30 ℃ for fermenting for 24 hours to obtain fermented hatching solution;
weighing 0.14g of aedes albopictus eggs, adding the eggs into a 50mL hatching tube, and adding 33.3mL of fermented hatching fluid (the fermented hatching fluid accounts for 2/3 of the volume of the hatching tube) into the hatching tube; sealing the tube cover of the incubation tube, shaking up, standing and incubating for 1 hour at the temperature of 25-30 ℃, then opening the tube cover of the incubation tube, and transferring the incubated larvae to a feeding tray.
Comparative example 3
The embodiment provides a hatching method of aedes albopictus eggs, which comprises the following steps:
adding water into yeast peptone to prepare an aqueous solution with the concentration of 0.9g/L, and taking the aqueous solution as hatching fluid;
placing the hatching solution at the temperature of 25-30 ℃ for fermenting for 24 hours to obtain fermented hatching solution;
weighing 0.14g of aedes albopictus eggs, adding the eggs into a 50mL hatching tube, and adding 15mL of fermented hatching fluid into the hatching tube (the fermented hatching fluid accounts for 30% of the volume of the hatching tube); sealing the tube cover of the incubation tube, shaking up, standing and incubating for 1 hour at the temperature of 25-30 ℃, then opening the tube cover of the incubation tube, and transferring the incubated larvae to a feeding tray.
Comparative example 4
The embodiment provides a hatching method of aedes albopictus eggs, which comprises the following steps:
adding water into yeast peptone to prepare an aqueous solution with the concentration of 0.9g/L, and taking the aqueous solution as hatching fluid;
placing the hatching solution at the temperature of 25-30 ℃ for fermenting for 24 hours to obtain fermented hatching solution;
weighing 0.14g of aedes albopictus eggs, adding the eggs into a 50mL hatching tube, and adding 47.5mL of fermented hatching fluid into the hatching tube (the fermented hatching fluid accounts for 95% of the volume of the hatching tube); sealing the tube cover of the incubation tube, shaking up, standing and incubating for 1 hour at the temperature of 25-30 ℃, then opening the tube cover of the incubation tube, and transferring the incubated larvae to a feeding tray.
The results of statistics of hatchability and larval survival rates of the active mosquito eggs of examples 1 to 5 and comparative examples 1 to 4 are shown in table 1 below, from which it can be seen that: the concentration of the yeast peptone in the hatching fluid and the addition of the hatching fluid in the hatching tube are all decisive for the hatching rate and the survival rate of the larvae. Wherein, the effect of the examples 1-5 is much better than that of the comparative examples 1-4, and the effect of the examples 1-3 is the best, and the hatchability of active mosquito eggs and the survival rate of larvae are high; example 4 incubation rate of active mosquito eggs was lower than examples 1-3 due to lower concentration of yeast peptone in hatching fluid, but did not affect larval survival rate; example 5 incubation rate of active mosquito eggs was high due to higher concentration of yeast peptone in the hatching fluid, but survival rate of hatched larvae was lower than that of examples 1-3. Comparative example 1 the concentration of yeast peptone in the hatching fluid was too high compared to example 1, resulting in very low survival of hatched larvae; comparative example 2 the concentration of yeast peptone in the hatching fluid was too low compared to example 1, resulting in an extremely low hatching rate of active mosquito eggs; compared with example 1, the addition amount of the hatching fluid is too small, so that the hatching rate of active mosquito eggs is low, and the survival rate of hatched larvae is low; comparative example 4 the hatching fluid was added in too much amount, resulting in low survival rate of larvae, compared to example 1.
TABLE 1
| Hatchability of active mosquito eggs | Survival rate of larvae | |
| Example 1 | 98.34％ | 99.32％ |
| Example 2 | 95.28％ | 99.15％ |
| Example 3 | 97.16％ | 99.43％ |
| Example 4 | 85.67％ | 99.28％ |
| Example 5 | 98.11％ | 74.92％ |
| Comparative example 1 | 98.39％ | 3.51％ |
| Comparative example 2 | 12.53％ | 99.19％ |
| Comparative example 3 | 35.56％ | 83.79％ |
| Comparative example 4 | 96.45％ | 37.07％ |
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. The hatching solution for hatching the mosquito eggs of aedes albopictus is characterized by being obtained by fermenting an aqueous solution prepared from yeast peptone and water, wherein the concentration of the yeast peptone in the aqueous solution is 0.7-1.5 g/L.
2. Hatching fluid for hatching aedes albopictus eggs according to claim 1, wherein the concentration of yeast peptone in the aqueous solution is 0.8-1.0 g/L.
3. Hatching fluid for hatching aedes albopictus eggs according to claim 2, wherein the concentration of yeast peptone in the aqueous solution is 0.9 g/L.
4. Hatching fluid for hatching aedes albopictus eggs according to any one of claims 1 to 3, wherein the yeast peptone is Angel yeast peptone.
5. An incubation method of aedes albopictus eggs is characterized by comprising the following steps:
obtaining a hatching fluid for hatching the eggs of aedes albopictus according to any one of claims 1 to 4;
weighing aedes albopictus eggs, adding the aedes albopictus eggs into an incubation tube, adding the incubation liquid into the incubation tube, sealing a tube cover of the incubation tube, shaking uniformly, standing for incubation, opening the tube cover of the incubation tube after incubation is finished, and transferring incubated larvae into a feeding tray.
6. An incubation method of aedes albopictus eggs according to claim 5, wherein the temperature of the incubation liquid is 25 ℃ to 30 ℃, and the time of the fermentation is 20 hours to 28 hours.
7. A method of incubating Aedes albopictus mosquito eggs as claimed in claim 5, wherein said hatching fluid is added in an amount of 50-80% of the volume of said hatching tube.
8. A method of incubating Aedes albopictus mosquito eggs as claimed in claim 7, wherein said hatching fluid is added in an amount of 60-70% of the volume of said hatching tube.
9. An incubation method of aedes albopictus eggs according to any one of claims 5-8, wherein the ratio of aedes albopictus eggs to the incubation solution is 1 g: 150 and 350 mL.
10. The method for hatching Aedes albopictus eggs as claimed in any one of claims 5 to 8, wherein the temperature of the static hatching is 25 ℃ to 30 ℃ and the time of the static hatching is 0.8 to 1.2 hours.
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| US8844465B2 (en) * | 2009-04-08 | 2014-09-30 | Kristian J. K. Holland | Apparatus and method for controlling maturation of aquatically hatched insects |
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