阅读说明：本技术 一种大量快速繁殖果蝇并获取幼虫的方法 (Method for rapidly breeding fruit flies in large quantity and obtaining larvae ) 是由 张威 张文琪 袁潮海 于 2021-09-23 设计创作，主要内容包括：本发明涉及一种大量快速繁殖果蝇并获取幼虫的方法,采用圆柱形、透明的培养管作为培养容器,管盖上设有通气的小孔,将培养管洗净后晾干,装入果蝇饲料,以(1-3)：1的数量比加入雌雄果蝇,培养12h后收集虫卵,用另一个新装有果蝇饲料的培养管扣在产有虫卵的培养管上,待果蝇全部爬到新培养管后,取下培养管,将新培养管盖上盖子后继续培养收集虫卵,将装有虫卵的旧培养管盖上盖子后进行孵育,3-4天后,将培养管中的果蝇饲料转移至广口容器中,再将容器置于水浴中,刮取食物表面的幼虫,将幼虫清洗后过筛,再去掉幼虫表面的水分。该方法可在短时间内获取大量干净、活力大的幼虫,大大缩短了以往果蝇培育的时间,降低了果蝇饲养成本。(The invention relates to a method for breeding fruit flies and obtaining larvae rapidly in large quantity, which adopts a cylindrical and transparent culture tube as a culture container, a tube cover is provided with a small hole for ventilation, the culture tube is cleaned and dried, and fruit fly feed is filled in the culture tube, wherein the ratio of the culture tube to the culture tube is (1-3): 1, culturing for 12 hours, collecting ova, covering another culture tube with a fruit fly feed on the culture tube for producing the ova, taking down the culture tube after the fruit flies all climb to the new culture tube, covering the new culture tube with a cover, continuously culturing and collecting the ova, covering the old culture tube with the ova with a cover, incubating, transferring the fruit fly feed in the culture tube to a wide-mouth container after 3-4 days, placing the container in a water bath, scraping larvae on the surface of the food, cleaning the larvae, sieving, and removing the moisture on the surface of the larvae. The method can obtain a large amount of clean and active larvae in a short time, greatly shorten the previous fruit fly cultivation time, and reduce the fruit fly breeding cost.)

1. A method for rapidly propagating drosophila melanogaster in large quantities and obtaining larvae is characterized by comprising the following steps:

(1) a cylindrical culture tube is used as a culture container, the inner diameter of the culture tube is 4-6cm, the height of the culture tube is 7-9cm, the culture tube is made of transparent material, a plurality of small ventilation holes are formed in a tube cover, and the culture tube is cleaned and dried for later use;

(2) loading fruit fly feed into culture tube, selecting young fruit fly within 24h of emergence, and making use of CO₂After anesthesia, picking out and removing residual wings and wingless fruit flies, selecting male and female fruit flies according to the ratio of male and female fruit flies (1-3): 1, adding the mixture into a culture tube, and then putting the culture tube into an incubator for culture;

(3) collecting ova after culturing for 12h, during collection, forcibly shaking the culture tube to enable the ova to sink to the bottom of the culture tube, then quickly opening a tube cover, buckling another culture tube newly filled with fruit fly feed on the culture tube with the ova, taking down the culture tube by utilizing the characteristic that fruit flies climb upwards to the wall, covering the new culture tube with the fruit flies with a cover, putting the new culture tube into an incubator for culturing, collecting the ova every 12h, covering the old culture tube with the ova with the cover, and transferring the old culture tube to the incubator for incubation;

(4) after incubating the old culture tube with the eggs for 3-4 days, gently taking out the feed of the fruit flies in the culture tube, transferring the feed to a wide-mouth container, placing the container in a water bath at 40-47 ℃, allowing the larvae to migrate to the surface of the feed of the fruit flies along with the rise of the temperature, scraping the larvae on the surface of the food, adding the larvae into the container filled with water, cleaning the larvae, sieving the larvae through a 80-mesh sieve, transferring the larvae to a culture dish paved with clean absorbent paper, and removing the water on the surfaces of the larvae to obtain the clean and moisture-free fruit fly larvae;

in the step (2), the fruit fly feed comprises the following raw materials: 2L of water, 55-65g of brown sugar, 14-16g of white sugar, 90-105g of non-degreased corn flour, 45-50g of live yeast powder, 15-18g of agar, 5-9ml of propionic acid and 35ml of nipagin solution; the nipagin solution is prepared according to the proportion that 1g of nipagin methyl ester is dissolved in 10ml of 95 v% ethanol solution.

2. The method for mass propagation of fruit flies and obtaining larvae according to claim 1, wherein in step (2), the fruit fly feed comprises the following raw materials: 2L of water, 60g of brown sugar, 14.5g of white sugar, 100g of non-degreased corn flour, 49g of live yeast powder, 16g of agar, 8ml of propionic acid and 35ml of nipagin solution.

3. The method for mass propagation of fruit flies and obtaining larvae according to claim 1, wherein in step (2), the ratio of the number of male and female fruit flies is 1.5: 1.

4. The method for mass production of fruit flies and obtaining larvae as claimed in claim 1, wherein in step (2), a layer of dried yeast powder is sprinkled on the surface of the fruit fly feed in the culture tube before the male and female fruit flies are added into the culture tube.

5. The method for mass propagation of fruit flies and harvesting of larvae according to claim 1, wherein in step (2), the culture conditions are: at 25 ℃, 60 to 70 percent of humidity and 12h/d of illumination.

6. A method for mass production of fruit flies and for harvesting larvae according to any one of claims 1 to 5, wherein in step (3) the incubation conditions are: 30 ℃, 60-70% of humidity and 12h/d of illumination.

Technical Field

The invention relates to a method for large-scale propagation of small insects, in particular to a method for large-scale rapid propagation of drosophila melanogaster and obtaining larvae.

Background

Fruit flies are insects of the order diptera, widely present in the temperate and tropical climatic regions of the world, often in human habitats, garbage dumps and rotted fruit surfaces. Fruit flies are vegetative saprophytic organisms which like to eat saccharomycetes. There are over 1000 species of drosophila that have been discovered so far. The life cycle of drosophila involves four phases: the life cycle of eggs, larvae, pupae and adults is short, and only about 10 days are needed from the eggs to the adult flies.

The drosophila larvae has the characteristics of high protein, high fat and the like, contains rich mineral elements, vitamins and chitin, can be used for extracting chitosan, and can even be developed into an edible insect for the food and feed industries. The drosophila melanogaster is small in size and strong in reproductive capacity, but has strong flying capacity and is difficult to capture, and the larvae of the drosophila melanogaster live in food and only migrate out of the food gradually when pupating, so that the mass acquisition of the larvae is limited, and the development of the drosophila melanogaster in the food and feed industries is influenced.

Disclosure of Invention

The invention aims to solve the defects of the prior art and provide a method for quickly breeding a large number of drosophila melanogaster and obtaining larvae, which can obtain a large number of drosophila melanogaster eggs in a short time and quickly obtain the larvae and can reduce the feeding cost of the drosophila melanogaster.

Technical scheme

A method for mass rapid propagation of Drosophila melanogaster and obtaining larvae comprises the following steps:

(1) a cylindrical culture tube is used as a culture container, the inner diameter of the culture tube is 4-6cm, the height of the culture tube is 7-9cm, the culture tube is made of transparent material, a plurality of small ventilation holes are formed in a tube cover, and the culture tube is cleaned and dried for later use;

(2) loading fruit fly feed into culture tube, selecting young fruit fly within 24h of emergence, and making use of CO₂After anesthesia, picking out and removing residual wings and wingless fruit flies, selecting male and female fruit flies according to the ratio of male and female fruit flies (1-3): 1 by number ratio, plusPutting the culture tube into a culture box for culture;

(3) collecting ova after culturing for 12h, during collection, forcibly shaking the culture tube to enable the ova to sink to the bottom of the culture tube, then quickly opening a tube cover, buckling another culture tube newly filled with fruit fly feed on the culture tube with the ova, taking down the culture tube by utilizing the characteristic that fruit flies climb upwards to the wall, covering the new culture tube with the fruit flies with a cover, putting the new culture tube into an incubator for culturing, collecting the ova every 12h, covering the old culture tube with the ova with the cover, and transferring the old culture tube to the incubator for incubation;

(4) after incubating the old culture tube with the eggs for 3-4 days, gently taking out the feed of the fruit flies in the culture tube, transferring the feed to a wide-mouth container, placing the container in a water bath at 40-47 ℃, allowing the larvae to migrate to the surface of the feed of the fruit flies along with the rise of the temperature, scraping the larvae on the surface of the feed, adding the larvae into the container filled with water, cleaning the larvae, sieving the larvae through a 80-mesh sieve, transferring the larvae to a culture dish paved with clean absorbent paper, and removing the moisture on the surfaces of the larvae to obtain the clean and moisture-free fruit fly larvae.

In the step (2), the fruit fly feed comprises the following raw materials: 2L of water, 55-65g of brown sugar, 14-16g of white sugar, 90-105g of non-degreased corn flour, 45-50g of live yeast powder, 15-18g of agar, 5-9ml of propionic acid and 35ml of nipagin solution; the nipagin solution is prepared according to the proportion that 1g of nipagin methyl ester is dissolved in 10ml of 95 v% ethanol solution.

Further, in the step (2), the fruit fly feed comprises the following raw materials: 2L of water, 60g of brown sugar, 14.5g of white sugar, 100g of non-degreased corn flour, 49g of live yeast powder, 16g of agar, 8ml of propionic acid and 35ml of nipagin solution.

Further, in the step (2), the ratio of the number of male and female fruit flies is 1.5:1, and if the number of male fruit flies is greater than that of female fruit flies in the same culture tube, the egg laying amount is greatly reduced, even when the ratio of male flies to female flies reaches 1: and 3, the female fruit flies stop laying eggs, and the optimal egg laying amount is reached when the number ratio of the female fruit flies to the male fruit flies is 1.5: 1.

Further, in the step (2), before the male and female fruit flies are added into the culture tube, a layer of dried yeast powder is scattered on the surface of the fruit fly feed in the culture tube, so that sufficient protein can be provided for the fruit flies, and the egg laying amount of the fruit flies is increased.

Further, in the step (2), the culture conditions are as follows: at 25 ℃, 60 to 70 percent of humidity and 12h/d of illumination.

Further, in the step (3), the incubation conditions are as follows: 30 ℃, 60-70% of humidity and 12h/d of illumination.

The invention has the beneficial effects that:

the invention relates to a method for breeding fruit flies and obtaining larvae rapidly in large quantities, which adopts a self-made fruit fly feed which can not only meet the nutritional requirements of the fruit flies for breeding in large quantities, but also is not easy to grow mould.

Detailed Description

The technical solution of the present invention is further illustrated by the following specific examples.

Example 1

A method for mass rapid propagation of Drosophila melanogaster and obtaining larvae comprises the following steps:

(1) a cylindrical culture tube is used as a culture container, the inner diameter of the culture tube is 5cm, the height of the culture tube is 7cm, the culture tube is made of transparent plastic, a plurality of small ventilation holes are formed in a tube cover, and the culture tube is cleaned and dried for later use;

(2) preparing a fruit fly feed, wherein the fruit fly feed comprises the following raw materials: 2L of water, 60g of brown sugar, 14.5g of white sugar, 100g of non-defatted corn flour, 49g of live yeast powder, 16g of agar, 8ml of propionic acid and 35ml of a nipagin solution, wherein the nipagin solution is prepared by dissolving 1g of nipagin methyl ester in 10ml of 95 v% ethanol solution;

the preparation method of the fruit fly feed comprises the following steps: adding 16g of agar powder into 1L of water, and heating and boiling to obtain an agar solution; and pouring 800ml of water into a 1L beaker, putting a rotor, carrying out magnetic stirring, sequentially adding brown sugar, white sugar, corn flour and yeast powder, stirring and dissolving, adding into the agar solution, washing the beaker with the last remaining 200ml of water, adding into the agar solution, stirring and mixing uniformly, adding propionic acid, mixing uniformly, cooling to 60 ℃, adding the nipagin solution, stirring and mixing uniformly to obtain the fruit fly feed.

The culture tubes were filled with fruit fly feed (20 ml of fruit fly feed per culture tube, 75 culture tubes in total), young fruit flies within 24 hours of emergence were selected, and the fruit flies were allowed to utilize CO₂After anesthesia, picking out the residual wings and the wingless fruit flies and removing, and selecting male and female fruit flies according to the ratio of 1.5:1, adding the mixture into culture tubes (90 female fruit flies and 60 male fruit flies in each culture tube), and before the female fruit flies and the male fruit flies are added into the culture tubes, scattering a layer of dry yeast powder on the surfaces of fruit fly feed in the culture tubes to provide sufficient protein for the fruit flies so as to increase the egg laying amount of the fruit flies; putting the culture tube into an incubator, and culturing at 25 ℃ under the conditions of 60-70% of humidity and 12h/d of illumination;

(3) collect the ovum after cultivateing 12h, during the collection, rock the culture tube hard, make the ovum sink to the culture tube bottom, then open the tube closure rapidly, detain on the culture tube of producing the ovum with another culture tube (treating the fruit fly fodder solidifies the back) that is equipped with the fruit fly fodder, utilize the fruit fly to climb up the characteristic of wall, treat that the fruit fly is whole to climb to the new culture tube after, take off the culture tube, cover 75 new culture tubes that are equipped with the fruit fly and put into the incubator behind the lid and cultivate, collect an ovum every 12h, cover 75 old culture tubes that are equipped with the ovum and transfer to the incubator after the lid and incubate, the incubation condition is: 30 ℃, 60-70% of humidity and 12h/d of illumination;

(4) after 4 days of incubation, observing that two or three larvae climb the wall of the tube wall, wherein the larvae have the strongest peristalsis ability, the larvae in the food can climb to the outer surface of the food when sensing the ambient temperature change, slightly taking out the food in the culture tube by using a spoon, paying attention to keep the integrity of the food shape so as not to damage the larvae, uniformly pouring the food into a 20cm x 10cm open-cover plastic container, putting each piece of food in turn to be beneficial to scraping, then placing the container into a 40-47 ℃ water bath kettle (the temperature range can ensure that the larvae can feel the ambient environment change and migrate to the surface of the food without damaging the activity and physiological state of the larvae), and exceeding the optimal survival temperature of the larvae with the continuous rise of the temperature of the food in the container, so that the larvae gradually climb out from the food to the surface, and a large amount of larvae gather into a hill-like pile on the surface of the food, scraping the larvae on the surface of the food from far to near by using a spoon, transferring the larvae to another container filled with water, manually shaking the container filled with pure water after scraping, separating food residues from the larvae to achieve the effect of cleaning the larvae, then sieving the larvae through a 80-mesh sieve, transferring the larvae to a culture dish paved with clean absorbent paper, removing the moisture on the surface of the larvae, and obtaining the clean and moisture-free drosophila larvae. In this example, 698g (fresh weight) of Drosophila larvae were collected from 75 old culture tubes containing eggs for a total consumption of 3L Drosophila feed. And (4) after the culture tube filled with the fruit flies in the step (3) is cultured for 12 hours, continuously collecting worm eggs until the fruit flies do not lay eggs any more.