CN104186432B - Cultivating method of nicotine-resisting black soldier fly strain - Google Patents
Cultivating method of nicotine-resisting black soldier fly strain Download PDFInfo
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- CN104186432B CN104186432B CN201410421012.1A CN201410421012A CN104186432B CN 104186432 B CN104186432 B CN 104186432B CN 201410421012 A CN201410421012 A CN 201410421012A CN 104186432 B CN104186432 B CN 104186432B
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- 241000709785 Hermetia illucens Species 0.000 title abstract 8
- SNICXCGAKADSCV-JTQLQIEISA-N (-)-Nicotine Chemical compound CN1CCC[C@H]1C1=CC=CN=C1 SNICXCGAKADSCV-JTQLQIEISA-N 0.000 claims abstract description 165
- 229960002715 nicotine Drugs 0.000 claims abstract description 165
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Abstract
The invention relates to the technical field of insect cultivating, in particular to a cultivating method of a nicotine-resisting black soldier fly strain. According to the cultivating method, black soldier fly ova of the nicotine-resisting black soldier fly strain are inoculated into first formula materials containing nicotine, the ova are cultivated until larvae are ripe and pupate, after black soldier fly pupae are subjected to emergence, a nicotine water solution is directly used for feeding adults, after the adults carry out oviposition, a next-generation domestication cultivating is carried out, and by continuous domestication cultivating of 20-30 generations, the stable nicotine-resisting black soldier fly strain can be obtained. Second formula materials are a water solution with the nicotine weight percent concentration of 0.2%-0.3%. According to the method, the nicotine water solution with the proper concentration is directly used for being combined with bran and imported fish meal in a scientific mode, the obtained first formula materials successfully cover the unpleasant smell and bitter taste of nicotine, nutrient components are proper, smooth growing of the black soldier fly larvae is guaranteed, due to the proper and certain nicotine content of the first formula materials and the second formula materials, the fact that the obtained black soldier fly ripe larva rates of the formula materials with the same weight combination in different batches under the same cultivation environment are basically the same is guaranteed, and a stable cultivating effect is obtained.
Description
Technical field
The present invention relates to insecticide Cultivating techniques field, and in particular to a kind of breeding method of the black soldier flies of resistance to nicotine strain.
Background technology
Inapplicable fresh tobacco leaf refers to the fresh tobacco leaves for not being suitable for cured tobacco production（Water content 80~90%）, it is big with document
The waste/hypo-tobacco leaf produced in the production process of flue-cured tobacco that amount is referred to or discarded tobacco leaf（Water content about 15%）It is different.Inapplicable Nicotiana tabacum L. leads to
Refer to cigarette strain normally to bind and remove after 2~3 milk foot leaves, cigarette industry it is inapplicable, need to be in the case where field be removed and is processed
Portion's leaf and upper leaf, mainly include cigarette strain bottom because illumination deficiency, dysplasia, inclusions are not enough, estimated roasting rear inferior quality,
Do not possess bad hacking at the top of 2 inferior leads of baking value, and cigarette strain, curtailment, structure is stiff, bake rear inferior quality
2 tops.
If these inapplicable Nicotiana tabacum Lves arbitrarily abandon field, environmental pollution and the repetition sense of tobacco viruses are easily caused
Dye, the processing mode destroyed according to the concentration of prior art or fill then needs to spend substantial amounts of expense, and current industry is per ton not
The processing cost mean height of Nicotiana tabacum L. is suitable for up to 200 yuan, each Nicotiana tabacum L. producing region is generally required under the organization leadership of local government of party committee,
Policy propaganda, tabulation registration are carried out, treatment facility is built, unified removed, record archives, publicity, cashes and the measure such as subsidize and supervise
The process of inapplicable Nicotiana tabacum L. is carried out, is wasted time and energy costly.
It is a kind of preferable alternate process method that biological treatment is carried out to inapplicable Nicotiana tabacum L. using the insecticide of resistance to nicotine, ring
Protect, high financial profit.In order to realize the inapplicable fresh tobacco leaf of biological treatment using insecticide, the insecticide product for filtering out resistance to nicotine are needed first
Kind.
The applicant is summed up using inapplicable Nicotiana tabacum L. based on studying for a long period of time to housefly and inapplicable Nicotiana tabacum L. treatment technology
Screening, reinforcing and the breeding method of resistance to nicotine strain are carried out to housefly.The cultivation is that the ovum of resistance to nicotine housefly is inoculated in into the
One formulation material, family's fly pupa feeds adult after sprouting wings with the second formulation material, and through the continuous domestication in 20~30 generations resistance to nicotine man is obtained
Fly kind.The composition that it is critical only that the first formulation material and the second formulation material of its method is ensureing the smooth life of resistance to nicotine housefly
It is long.First formulation material consists of inapplicable fresh tobacco leaf 75~80%, wheat bran 10~15%, imported fish meal 5~10%；Second formulation material
Consist of whole milk powder 70%, brown sugar 29.8~29.9%, pure nicotine 0.1~0.2%.
The housefly kind that the technical scheme can go out resistance to nicotine with successful incubation, but different batches cultivate the aging black of acquisition
The quantity difference of stratiomyiid larva is larger, and resistance to nicotine ability is also otherwise varied, there is the uncertainty of effect.Main reason is that not
It is suitable for the application aspect of Nicotiana tabacum L., although sum up through creative research and many experiments and add in formulation material zone of reasonableness
Inapplicable Nicotiana tabacum L. can ensure the control to Musca domestica larva toxic action, but affect the factor of not nicotine content of tobacco leaves very various,
Complicated mechanism, it is more difficult to realize accurately control.Affecting the factor of nicotine content in tobacco leaf mainly includes：（1）Different growing stages, Nicotiana tabacum L.
In contained nicotine content it is different：In tobacco seed nicotine content be 0, rise in seedling raising process beginning formed in root, after with root
System develops, and the synthesis of nicotine and accumulation ability gradually strengthen, so different growing stages, contained nicotine content is not in Nicotiana tabacum L.
Together；（2）Cigarette strain different parts, nicotine content is dramatically different, and nicotine content is also by leaf position and number of sheets large effect；（3）It is different
Nicotine amount contained by kind Nicotiana tabacum L. is different；（4）Weather conditions of soil dry-wet etc. have the impact of different savings to nicotine content；
（5）The different appreciable impact nicotine content of tobacco leaves of agronomic measures, including fertilization type, planting density, pinch smear wooden fork fertilising, girdling
With cut the nicotine content in root fertilizer, picking methods etc. factor all appreciable impact Nicotiana tabacum Lves, by taking fertilization type as an example, three kinds of forms
Nitrogenous fertilizer in, carbamide apply after, nicotine accumulation highest, ammonium nitrate takes second place, potassium nitrate process nicotine accumulation it is minimum, than apply carbamide
It is low by 27%；By taking Physical property as an example, the research such as Zhang Dan finds, postpones nicotine accumulation intensity peak with Physical property and declines successively, cigarette
Alkali content also lowers successively, often postpones and pinching for one day, and nicotine content just declines 0.0504%；Again to cut root fertilizer as a example by, cut root 1/
8 compared with root is not cut, and upper leaf nicotine declines 1.9 percentage points of ranges of decrease 31.33%, and middle leaf nicotine declines 2.13 percentage points,
The range of decrease 51.4%.How to avoid the uncertainty of the aging black soldier flies larva effect for cultivating acquisition using different batches compost is
This area technical barrier urgently to be resolved hurrily.And adopt and stablize the resistance to nicotine insecticide product that feasible technical scheme cultivates more multi items
It is and effectively continuous domestication is stabilized it, is this area important technological problems that simultaneously needs are solved.
The content of the invention
The technical problem to be solved in the present invention be in order to overcome prior art in lack the defect of the black soldier flies of resistance to nicotine kind, carry
For a kind of breeding method of the black soldier flies of resistance to nicotine strain, the inventive method is not affected by inapplicable Nicotiana tabacum L. various factors, can be obtained
Stable cultivate effect.
The purpose of the present invention is achieved by the following technical programs：
Black soldier flies are a kind of insecticides for being different from housefly, and it is a kind of important economic insects and scavenger insect, available
The various corrupt organic wastes of black soldier flies process, such as beer residue, bean dregs, chicken manure, pig manure and changing food waste.Existing black soldier flies kind
There are preferable biological treatment capabilities to nontoxic organic waste, but it is not good to not applying to the treatment effect of fresh tobacco leaf.Mainly
Reason is because in inapplicable fresh tobacco leaf containing toxicants such as a certain amount of nicotine, to black soldier flies larva（The aging children of black soldier flies
Worm）Effect with certain murder by poisoning and Developing restraint.In order to using the inapplicable fresh tobacco leaf of black soldier flies biological treatment, first need to be from
The black soldier flies kind of resistance to nicotine is filtered out in wild black soldier flies kind, tolerance of the black soldier flies to fresh tobacco leaf is improved.
The present invention provides a kind of breeding method of the black soldier flies of resistance to nicotine strain, comprises the steps：
S1. the black soldier flies kind of the nicotine aqueous formulation material that mass percent concentration is 3 ‰ will be tolerated under indoor conditionss
Ovum be seeded in the first formulation material, by worm's ovum cultivate to larva it is aging, pupate；
S2. black soldier flies pupa feeds its adult after sprouting wings using the second formulation material, after its spawning, into taming and dociling for next generation
Change and cultivate；
S3. above-mentioned steps repetitive operation is pressed, is cultivated through the continuous domestication of 20~30 generations, obtain stable resistance to nicotine black
Stratiomyiid strain；
Wherein, first formulation material is grouped into by each group of following percetage by weight：Wheat bran 70~90%, imported fish meal
10~30%, above-mentioned formulation material is adjusted to into humidity for 70% with the nicotine aqueous solution of mass percent concentration 3 ‰；
Second formulation material is the aqueous solution of nicotine mass percent concentration 2 ‰~3 ‰.
Preferably, first formulation material is grouped into by each group of following percetage by weight：Wheat bran 80%, imported fish meal
20%, above-mentioned formulation material is adjusted to into humidity for 70% with the nicotine aqueous solution of mass percent concentration 3 ‰.
Preferably, the second formulation material is the aqueous solution of nicotine mass percent concentration 2.5 ‰.
The nicotine can adopt the commercial products of purity >=99.5%.
The compound method of first formulation material is after each component mixing by the first formulation material, to add extra fine quality percentage
The humidity of formulation material is adjusted to 70% by the nicotine aqueous solution of specific concentration.
The g of black soldier flies ovum 0.5~1.0 is accessed among every formulation materials of 1 kg first.It is preferred that every formula of 1 kg first
The g of black soldier flies ovum 0.8 is accessed in material.
The tolerance mass percent concentration is that the black soldier flies of the resistance to nicotine kind of 3 ‰ nicotine aqueous formulation material refers to head
First the black soldier flies of resistance to nicotine kind is filtered out from wild black soldier flies kind, then again by the black soldier flies of the resistance to nicotine kind for screening
Carry out hot housing so as to tolerate the nicotine aqueous formulation material that mass percent concentration is 3 ‰.
It is described the black soldier flies of resistance to nicotine kind is filtered out from wild black soldier flies kind concrete grammar be：
S01. configuration traps compost, will trap compost and is fitted in open containers；
It is described to trap each component that compost contains following percetage by weight：Wheat bran 70~90%, imported fish meal 10~30%,
Above-mentioned formulation material is adjusted to into humidity for 70% with the nicotine aqueous solution of mass percent concentration 1 ‰~2 ‰；
S02. when weather is fine, the morning 8:00~9:00 container that will be equipped with trapping compost is put in respectively house refuse
Heap, chicken house and pig farm are trapping wild worm's ovum, afternoon 17:00~18:00 withdraws the container being put in everywhere；
S03. by reclaim each container be placed on indoor cultivation, wait the wild worm's ovum for trapping cultivate to larva it is aging when, profit
With black soldier flies mature larva black soldier flies larva is separated with formulation material from food phase behavior；
S04. by the Morphological Identification of black soldier flies larva, black soldier flies mature larva is chosen；
S05. filter out that 20~25 days aging, every black soldier flies larva weighs 0.10~0.16 g, figure is plump, color is pitch-black
Black soldier flies mature larva, at ambient temperature, allow black soldier flies mature larva to pupate naturally, sprout wings after by obtain resistance to nicotine
Black soldier flies kind.
It is described that the black soldier flies of resistance to nicotine for screening kind is carried out into hot housing so as to which that tolerating mass percent concentration is
The concrete grammar of 3 ‰ nicotine aqueous formulation material is：
S11. starting stage：The ovum of the black soldier flies of resistance to nicotine kind is seeded in initial incubation material, the initial incubation material
It is grouped into by each group of following percetage by weight：Wheat bran 70~90%, imported fish meal 10~30%, with mass percent concentration 1 ‰
Nicotine aqueous solution above-mentioned formulation material is adjusted to into humidity for 70%；In room conditions, by above-mentioned worm's ovum cultivate to larva it is aging,
After pupating, sprouting wings, its adult is fed；
S12. the 1st strain：After the Adult worms producting eggs of S11, the ovum for being produced is entered using the 1st hot housing material
Row cultivates culture；The 1st hot housing material contains each component of following percetage by weight：Wheat bran 70~90%, imported fish meal
10~30%, use mass percent concentration（1+S）Above-mentioned formulation material is adjusted to ‰ nicotine aqueous solution the value that humidity is 70%, S
Selected from 0.1~0.5；In room conditions, by above-mentioned worm's ovum cultivate to larva it is aging, pupate, sprout wings after, feed its adult；
S13. the N+1 time strain：Hot housing is carried out again to the ovum that the adult obtained by n-th strain produces,
And component contained by the compost used by the N+1 time strain meets following ratio：The mass percent of nicotine aqueous solution is dense
Degree=（N-th cultivates the content+S of nicotine in compost）‰, remaining constituent content is constant, and N is the natural number more than or equal to 1, weight
Multiple reinforcing, the mass percent concentration of nicotine aqueous solution is up to till 3 ‰ in the compost described in last time breeding phase.
Described its adult of nursing is fed using clear water.Preferably, the clear water may be selected pure water, tap water, well
Water or nontoxic river etc.；The mode that water is fed is to be saturated with water using sponge, is sucked for adult.
Preferably, it is described cultivate black soldier flies indoor conditionss be 25~30 DEG C of temperature, humidity 30~70%.
The invention has the beneficial effects as follows：
The present invention creatively directly using the nicotine aqueous solution and wheat bran and imported fish meal scientific compatibility of suitable concentration, is obtained
The first formulation material for obtaining not only successfully covers the unpleasant and bitterness of nicotine, and nutritional labeling is appropriate, it is ensured that black soldier flies larval growth is suitable
Profit, the and because appropriateness and definitiveness of the nicotine content of the first formulation material and the second formulation material, and the first formulation material and
The reasonability arranged before and after second formulation material nutrient substance and nicotine content, it is ensured that in same nurturing an environment, different batches
Same weight composition the black soldier flies mature larva rate that obtains of formulation material it is essentially identical, obtain stable cultivate effect.
Through the present invention black soldier flies of the resistance to nicotine strain for obtaining is cultivated due to have passed through the long-term acclimatization culture in 20~30 generations,
Culture conditions are not only adapted to, and has defined more stable hereditary character, the stability of the resistance to nicotine ability of its offspring is very
It is good, and the black soldier flies of the resistance to nicotine strain for obtaining is cultivated without the present invention（Only through screening and the black soldier flies of hot housing）Offspring
The toleration of nicotine is easily degenerated, stability is bad.
Specific embodiment
The present invention is further described with reference to specific embodiment.Unless stated otherwise, the present invention adopt reagent,
Equipment is the art conventional reagent and equipment.
Nicotine is purchased from conventional chemical reagent shop, and purity is >=99.5%.
Embodiment 1 filters out the black soldier flies of resistance to nicotine kind from wild black soldier flies kind
S01. when weather is fine, preparation traps compost, each component for trapping compost by following percetage by weight
Composition：Wheat bran 90%, imported fish meal 10%, humidity is adjusted to the nicotine aqueous solution of mass percent concentration 1 ‰ by above-mentioned formulation material
For 70%.
S02. by the above-mentioned compost for modulating, every 1 kg is fitted in a square plastic feed basin（Long * width * height=45cm*30cm*
12cm）.In morning 8 on the same day:00 plastic tub that will be equipped with compost is put in by a life rubbish heap, afternoon 17:00 withdraws everywhere
Plastic tub, takes back indoor cultivation.
S03. in room conditions（25 DEG C of temperature, humidity 30%）, the wild worm's ovum for trapping is cultivated into aging to larva,
Using black soldier flies mature larva black soldier flies larva is separated with formulation material from food phase behavior（When i.e. black soldier flies mature larva is aging
Can voluntarily climb from compost, find and pupated where being dried）.
S04. black soldier flies mature larva is individually chosen with tweezers by the Morphological Identification of larva under anatomical lens, is excluded
Other fly kinds.
S05. filter out that 20~25 days aging, every black soldier flies larva weighs 0.10~0.16 g, figure is plump, color is pitch-black
Black soldier flies mature larva, at ambient temperature, allow these black soldier flies mature larvas to pupate naturally, sprout wings after by obtain resistance to cigarette
The black soldier flies kind of alkali.
Embodiment 2 filters out the black soldier flies of resistance to nicotine kind from wild black soldier flies kind
S01. when weather is fine, preparation traps compost, each component for trapping compost by following percetage by weight
Composition：Wheat bran 80%, imported fish meal 20%, are adjusted to above-mentioned formulation material with the nicotine aqueous solution of mass percent concentration 1.5 ‰ wet
Spend for 70%.
S02. by the above-mentioned compost for modulating, it is fitted in a square plastic feed basin per 1kg（Long * width * height=45cm*30cm*
12cm）.In morning 8 on the same day:00 plastic tub that will be equipped with compost is put in by a life rubbish heap, afternoon 17:00 withdraws everywhere
Plastic tub, takes back indoor cultivation.
S03. in room conditions（25 DEG C of temperature, humidity 30%）, the wild worm's ovum for trapping is cultivated into aging to larva,
Using black soldier flies mature larva black soldier flies larva is separated with formulation material from food phase behavior（When i.e. black soldier flies mature larva is aging
Can voluntarily climb from compost, find and pupated where being dried）.
S04. black soldier flies mature larva is individually chosen with tweezers by the Morphological Identification of larva under anatomical lens, is excluded
Other fly kinds.
S05. filter out that 20~25 days aging, every black soldier flies larva weighs 0.10~0.16 g, figure is plump, color is pitch-black
Black soldier flies mature larva, at ambient temperature, allow these black soldier flies mature larvas to pupate naturally, sprout wings after by obtain resistance to cigarette
The black soldier flies kind of alkali.
Embodiment 3 filters out the black soldier flies of resistance to nicotine kind from wild black soldier flies kind
S01. when weather is fine, preparation traps compost, each component for trapping compost by following percetage by weight
Composition：Wheat bran 70%, imported fish meal 30%, humidity is adjusted to the nicotine aqueous solution of mass percent concentration 2 ‰ by above-mentioned formulation material
For 70%.
S02. by the above-mentioned compost for modulating, every 1 kg is fitted in a square plastic feed basin（Long * width * height=45cm*30cm*
12cm）.In morning 8 on the same day:00 plastic tub that will be equipped with compost is put in by a life rubbish heap, afternoon 17:00 withdraws everywhere
Plastic tub, takes back indoor cultivation.
S03. in room conditions（25 DEG C of temperature, humidity 30%）, the wild worm's ovum for trapping is cultivated into aging to larva,
Using black soldier flies mature larva black soldier flies larva is separated with formulation material from food phase behavior（When i.e. black soldier flies mature larva is aging
Can voluntarily climb from compost, find and pupated where being dried）.
S04. black soldier flies mature larva is individually chosen with tweezers by the Morphological Identification of larva under anatomical lens, is excluded
Other fly kinds.
S5. filter out that 20~25 days aging, every black soldier flies larva weighs 0.10~0.16 g, figure is plump, color is pitch-black
Black soldier flies mature larva, at ambient temperature, allow these black soldier flies mature larvas to pupate naturally, sprout wings after by obtain resistance to cigarette
The black soldier flies kind of alkali.
The 4 pairs of black soldier flies of resistance to nicotine for screening kinds of embodiment carry out hot housing makes its tolerance mass percent dense
Spend the formulation material of the nicotine aqueous solution for 3 ‰
The black soldier flies of the resistance to nicotine kind that any embodiment of embodiment 1~3 is screened is cultivated so as to tolerate quality
Percent concentration is the formulation material of 3 ‰ nicotine aqueous solution, and concrete grammar is as follows：
S11. starting stage：
Initial incubation material is prepared, the initial incubation material is grouped into by each group of following percetage by weight：Wheat bran 90%, enter
Mouth fish flour 10%, humidity is adjusted to for 70% with the nicotine aqueous solution of mass percent concentration 1 ‰ by above-mentioned formulation material.By above-mentioned modulation
Good compost, every 1 kg is fitted in a square plastic feed basin（Long * width * height=45cm*30cm*12cm）, black soldier flies ovum is accessed per basin
0.8 g；In room conditions（25~30 DEG C of temperature, humidity 30~70%）, by above-mentioned worm's ovum cultivate to larva it is aging, pupate, plumage
After change, its adult is fed.
Strain：
S12. the 1st strain：After the Adult worms producting eggs of starting stage, the ovum for being produced is using the 1st reinforcing training
Nutriment carries out hot housing；The 1st hot housing material contains each component of following percetage by weight：Wheat bran 80%, import fish
Powder 20%, uses mass percent concentration（1+S）Above-mentioned formulation material is adjusted to humidity and is selected from 0.1 for 70%, S by ‰ nicotine aqueous solution
~0.5；In room conditions（25~30 DEG C of temperature, humidity 30~70%）, by above-mentioned worm's ovum cultivate to larva it is aging, pupate, plumage
After change, its adult is fed.
S13. the N+1 time strain：Hot housing is carried out again to the ovum that the adult obtained by n-th strain produces,
And component contained by the compost used by the N+1 time strain meets following ratio：The mass percent of nicotine aqueous solution is dense
Degree=（N-th cultivates the content+S of nicotine in compost）‰, remaining constituent content is constant, and N is the natural number more than or equal to 1, weight
Multiple reinforcing, the mass percent concentration of nicotine aqueous solution is up to till 3 ‰ in the compost described in last time strain.
Preferably, in this enforcement, in whole breeding phase, the mass percent concentration of nicotine aqueous solution is followed successively by
1.1‰、1.2‰、1.3‰、1.4‰、1.5‰、1.6‰、1.7‰、1.8‰、1.9‰、2.0‰、2.1‰、2.2‰、
2.3‰、2.4‰、2.5‰、2.6‰、2.7‰、2.8‰、2.9‰、3.0‰；Namely black soldier flies ovum experiences altogether 20 reinforcing trainings
Support.The black soldier flies kind that this stage turns out has higher tolerance to nicotine.
The 5 pairs of black soldier flies of resistance to nicotine for screening kinds of embodiment carry out hot housing makes its tolerance mass percent dense
Spend the formulation material of the nicotine aqueous solution for 3 ‰
The black soldier flies of the resistance to nicotine kind that any embodiment of embodiment 1~3 is screened is cultivated so as to tolerate quality
Percent concentration is the formulation material of 3 ‰ nicotine aqueous solution, and concrete grammar is as follows：
S11. starting stage：
Initial incubation material is prepared, the initial incubation material is grouped into by each group of following percetage by weight：Wheat bran 80%, enter
Mouth fish flour 20%, humidity is adjusted to for 70% with the nicotine aqueous solution of mass percent concentration 1 ‰ by above-mentioned formulation material.By above-mentioned modulation
Good compost, every 1 kg is fitted in a square plastic feed basin（Long * width * height=45cm*30cm*12cm）, black soldier flies ovum is accessed per basin
0.8 g；In room conditions（25~30 DEG C of temperature, humidity 30~70%）, by above-mentioned worm's ovum cultivate to larva it is aging, pupate, plumage
After change, its adult is fed.
Strain：
S12. the 1st strain：After the Adult worms producting eggs of starting stage, the ovum for being produced is using the 1st reinforcing training
Nutriment carries out hot housing；The 1st hot housing material contains each component of following percetage by weight：Wheat bran 80%, import fish
Powder 20%, uses mass percent concentration（1+S）‰ nicotine aqueous solution by above-mentioned formulation material be adjusted to humidity for 70%, S selected from 0.1~
0.5；In room conditions（25~30 DEG C of temperature, humidity 30~70%）, by above-mentioned worm's ovum cultivate to larva it is aging, pupate, sprout wings
Afterwards, its adult is fed；
S13. the N+1 time strain：Hot housing is carried out again to the ovum that the adult obtained by n-th strain produces,
And component contained by the compost used by the N+1 time strain meets following ratio：The mass percent of nicotine aqueous solution is dense
Degree=（N-th cultivates the content+S of nicotine in compost）‰, remaining constituent content is constant, and N is the natural number more than or equal to 1, weight
Multiple reinforcing, the mass percent concentration of nicotine aqueous solution is up to till 3 ‰ in the compost described in last time strain.
Preferably, in this enforcement, in whole breeding phase, the mass percent concentration of nicotine aqueous solution is followed successively by
1.2‰、1.4‰、1.6‰、1.8‰、2.0‰、2.2‰、2.4‰、2.6‰、2.8‰、3.0‰；Namely the common Jing of black soldier flies ovum
Go through 10 hot housing.The black soldier flies kind that this stage turns out has higher tolerance to nicotine.
The 6 pairs of black soldier flies of resistance to nicotine for screening kinds of embodiment carry out hot housing makes its tolerance mass percent dense
Spend the formulation material of the nicotine aqueous solution for 3 ‰
The black soldier flies of the resistance to nicotine kind that any embodiment of embodiment 1~3 is screened is cultivated so as to tolerate quality
Percent concentration is the formulation material of 3 ‰ nicotine aqueous solution, and concrete grammar is as follows：
S11. starting stage：
Initial incubation material is prepared, the initial incubation material is grouped into by each group of following percetage by weight：Wheat bran 70%, enter
Mouth fish flour 30%, humidity is adjusted to for 70% with the nicotine aqueous solution of mass percent concentration 1 ‰ by above-mentioned formulation material.By above-mentioned modulation
Good compost, every 1 kg is fitted in a square plastic feed basin（Long * width * height=45cm*30cm*12cm）, black soldier flies ovum is accessed per basin
0.8 g；In room conditions（25~30 DEG C of temperature, humidity 30~70%）, by above-mentioned worm's ovum cultivate to larva it is aging, pupate, plumage
After change, its adult is fed.Described its adult of nursing is fed using clear water.The clear water may be selected pure water, tap water, well
Water or nontoxic river etc.；The mode that water is fed is to be saturated with water using sponge, is sucked for adult.Similarly hereinafter.
Strain：
S12. the 1st strain：After the Adult worms producting eggs of starting stage, the ovum for being produced is using the 1st reinforcing training
Nutriment carries out hot housing；The 1st hot housing material contains each component of following percetage by weight：Wheat bran 80%, import fish
Powder 20%, uses mass percent concentration（1+S）Above-mentioned formulation material is adjusted to humidity and is selected from 0.1 for 70%, S by ‰ nicotine aqueous solution
~0.5；In room conditions（25~30 DEG C of temperature, humidity 30~70%）, by above-mentioned worm's ovum cultivate to larva it is aging, pupate, plumage
After change, its adult is fed；
S13. the N+1 time strain：Hot housing is carried out again to the ovum that the adult obtained by n-th strain produces,
And component contained by the compost used by the N+1 time strain meets following ratio：The mass percent of nicotine aqueous solution is dense
Degree=（N-th cultivates the content+S of nicotine in compost）‰, remaining constituent content is constant, and N is the natural number more than or equal to 1, weight
Multiple reinforcing, the mass percent concentration of nicotine aqueous solution is up to till 3 ‰ in the compost described in last time strain.
Preferably, in this enforcement, in whole breeding phase, the mass percent concentration of nicotine aqueous solution is followed successively by
1.5‰、2.0‰、2.5‰、3.0‰；Namely black soldier flies ovum experiences altogether 4 hot housing.The black soldier flies kind that this stage turns out
There is higher tolerance to nicotine.
The continuous domestication of more than 7 generation of embodiment is cultivated and obtains the stable black soldier flies of resistance to nicotine strain
The tolerance mass percent concentration that any embodiment of embodiment 4~6 is cultivated is that 3 ‰ nicotine aqueous solution is matched somebody with somebody
The black soldier flies kind of square stock is cultivated again through the continuous domestication of multiple generations, forms the stable black soldier flies of resistance to nicotine strain.Specifically
Method is：
S1. under indoor conditionss, the black soldier flies kind of the nicotine aqueous formulation material that mass percent concentration is 3 ‰ will be tolerated
Ovum be seeded in the first formulation material, by worm's ovum cultivate to larva it is aging, pupate；
S2. black soldier flies pupa feeds its adult after sprouting wings using the second formulation material, after its spawning, into taming and dociling for next generation
Change and cultivate；
S3. above-mentioned steps repetitive operation is pressed, is cultivated through the continuous domestication of 30 generations, obtain the stable black soldier flies of resistance to nicotine
Strain；
The preparation of the first formulation material：First formulation material is grouped into by each group of following percetage by weight：Wheat bran 90%,
Imported fish meal 10%, humidity is adjusted to for 70% with the nicotine aqueous solution of mass percent concentration 3 ‰ by above-mentioned formulation material.
The above-mentioned formulation material for modulating, every 1 kg are fitted in a square plastic feed basin（Long * width * height=45cm*30cm*
12cm）, the g of black soldier flies ovum 0.8 is accessed per basin；
In room conditions（25~30 DEG C of temperature, humidity 30~70%）, by above-mentioned worm's ovum cultivate to larva it is aging, pupate；
Black soldier flies pupa feeds its adult after sprouting wings using the second formulation material, and the second formulation material is nicotine mass percent concentration
2 ‰ aqueous solution, after its spawning, into the acclimatization culture of next generation；
By above-mentioned steps repetitive operation, cultivate through the continuous domestication of 30 generations, obtain the stable black soldier flies of resistance to nicotine product
System.
Embodiment 8
Other operations are grouped into embodiment 7, except for the difference that first formulation material by each group of following percetage by weight：
Wheat bran 80%, imported fish meal 20%, humidity is adjusted to for 70% with the nicotine aqueous solution of mass percent concentration 3 ‰ by above-mentioned formulation material.
Second formulation material is the aqueous solution of nicotine mass percent concentration 2.5 ‰.Cultivate through the continuous domestication of 25 generations, obtain
The stable black soldier flies of resistance to nicotine strain.
Embodiment 9
Other operations are grouped into embodiment 7, except for the difference that first formulation material by each group of following percetage by weight：
Wheat bran 70%, imported fish meal 30%, humidity is adjusted to for 70% with the nicotine aqueous solution of mass percent concentration 3 ‰ by above-mentioned formulation material.
Second formulation material is the aqueous solution of nicotine mass percent concentration 3 ‰.Cultivate through the continuous domestication of 20 generations, obtain steady
The fixed black soldier flies of resistance to nicotine strain.
Comparative example 1
Operation is grouped into embodiment 1, the except for the difference that compost that traps by each group of following percetage by weight：Wheat bran
80% and imported fish meal 20%.Experiment is found using as above trapping the kind that the ovum that compost traps is not resistance to nicotine.
Embodiment 10
Black soldier flies kind, the embodiment 4 to 6 that the embodiment of the present invention 1 to 3 is screened is through cultivating the cigarette for tolerating 3 ‰
Black soldier flies kind, the embodiment 7 to 9 of alkali formulation material is through the black soldier flies kind more for continuous domestication culture, the process of each comparative example
Black soldier flies kind afterwards carries out resistance to nicotine aptitude tests, and test process is：
The ovum 0.5g that the black soldier flies adult after the embodiment culture is processed is produced is taken respectively, is respectively connected to containing for 1kg weights
The formulation material of better quality percent concentration nicotine aqueous solution（Wheat bran 80%, imported fish meal 20%, use mass percent concentration
Above-mentioned formulation material is adjusted to humidity for 70% by 3.0 ‰ nicotine aqueous solution）In, test result is shown in Table 1：
Table 1
Test result illustrates that comparative example 1 is not resistant to the nicotine aqueous solution of mass percent concentration 3.0 ‰.
1 to 3 black soldier flies kind through screening of embodiment is modulated in the nicotine aqueous solution of mass percent concentration 3.0 ‰
Diapause phenomenon is there occurs in formulation material, larval weight is lighter, and have relatively large death.After the hot housing of embodiment 4 to 9 is processed
The resistance to nicotine ability of black soldier flies kind is greatly improved, and can tolerate the nicotine aqueous solution modulation of mass percent concentration 3.0 ‰
Formulation material.
The stability experiment of embodiment 11
The black soldier flies of the resistance to nicotine kind obtained through the methods described of embodiment 4 to 6 is carried out into conventional cultivation（Wherein black soldier flies are young
Worm culture material formula is wheat bran 80%+ fish flour 20%, and it is 70% that above-mentioned formulation material is adjusted to into humidity with clear water, and adult forage is clear water）
It is standby after 25 generations.Meanwhile, the black soldier flies of the resistance to nicotine strain that the methods described of Jing embodiments 7 to 9 is obtained is using cultivation material（It is wherein black
Stratiomyiid larva culture material formula be wheat bran 80%, imported fish meal 20%, will be upper with the nicotine aqueous solution of mass percent concentration 3.0 ‰
It is 70% to state formulation material and be adjusted to humidity, and adult forage is the nicotine aqueous solution of mass percent concentration 2.5 ‰）25 are persistently cultivated again
For rear standby.The black soldier flies kind of above-mentioned two strain is carried out into resistance to nicotine aptitude tests, detection culture material formula is wheat bran
80%th, imported fish meal 20%, be by the humidity that above-mentioned formulation material is adjusted to formulation material with the nicotine aqueous solution of mass percent concentration 3 ‰
70%, whether the resistance to nicotine ability for detecting its offspring declines.Test result is shown in Table 2：
The stability experiment result of black soldier flies strain of the table 2 after 25 generations
| Process | Larvae development goes through the phase（My god） | Larva bar number/g（Bar） | Survival rate of larvae（%） |
| The conventional strain of embodiment 4 | 7 | 48 | 63 |
| The conventional strain of embodiment 5 | 7 | 47 | 66 |
| The conventional strain of embodiment 6 | 7 | 48 | 64 |
| Embodiment 7 cultivates strain | 5 | 28 | 96 |
| Embodiment 8 cultivates strain | 5 | 28 | 96 |
| Embodiment 9 cultivates strain | 5 | 29 | 95 |
As a result the black soldier flies of the resistance to nicotine kind after reinforcing is shown after with the generation of cellar culture material culture 25, the energy of resistance to nicotine
Power there occurs serious degradation phenomenon, and pass through that the present invention cultivates that the black soldier flies of the resistance to nicotine lines progenies for obtaining tolerate to nicotine it is steady
It is qualitative fine, and in inventive formulation material compositing range, the Heisui River of the formulation material acquisition of the same weight composition of different batches
Horsefly has stable cultivate effect.
Claims (8)
1. a kind of breeding method of the black soldier flies of resistance to nicotine strain, it is characterised in that comprise the steps：
S1. under indoor conditionss, the ovum of the black soldier flies kind of the nicotine aqueous formulation material that mass percent concentration is 3 ‰ will be tolerated
In being seeded in the first formulation material, by worm's ovum cultivate to larva it is aging, pupate；
S2. fly pupa feeds its adult after sprouting wings using the second formulation material, after its spawning, into the acclimatization culture of next generation；
S3. above-mentioned steps repetitive operation is pressed, is cultivated through the continuous domestication of 20~30 generations, obtain the stable black soldier flies of resistance to nicotine
Strain；
Wherein, first formulation material is grouped into by each group of following percetage by weight：Wheat bran 80%, imported fish meal 20%, use
Above-mentioned formulation material is adjusted to humidity for 70% by the nicotine aqueous solution of mass percent concentration 3 ‰；
Second formulation material is the aqueous solution of nicotine mass percent concentration 2 ‰~3 ‰.
2. breeding method according to claim 1, it is characterised in that indoor conditionss described in S1 are 25~30 DEG C of temperature, wet
Degree 30~70%.
3. breeding method according to claim 1, it is characterised in that second formulation material is nicotine mass percent concentration
2.5 ‰ aqueous solution.
4. breeding method according to claim 1, it is characterised in that the compound method of first formulation material is to match somebody with somebody first
After each component of square stock is mixed, above-mentioned formulation material is adjusted to into humidity for 70% with the nicotine aqueous solution of mass percent concentration 3 ‰.
5. breeding method according to claim 1, it is characterised in that black soldier flies ovum 0.5 is accessed among first formulation material
The formulation materials of~1.0g/1kg first.
6. breeding method according to claim 1, it is characterised in that the tolerance mass percent concentration is 3 ‰ nicotine
The preparation method of the black soldier flies kind of aqueous formulation material is：Filter out the black soldier flies of resistance to nicotine from wild black soldier flies kind first
Kind, then by the black soldier flies of the resistance to nicotine kind for screening carries out hot housing so as to which tolerating mass percent concentration is again
3 ‰ nicotine aqueous formulation material.
7. breeding method according to claim 6, it is characterised in that the method for the screening of the black soldier flies of the resistance to nicotine kind
For：
S1. configuration traps compost, will trap compost and is fitted in open containers；
It is described to trap each component that compost contains following percetage by weight：Wheat bran 70~90%, imported fish meal 10~30%, use
The above-mentioned compost that traps is adjusted to humidity for 70% by the nicotine aqueous solution of mass percent concentration 1 ‰~2 ‰；
S2. when weather is fine, will be equipped with trapping the container of compost and be put in house refuse heap, chicken house and pig farm respectively to lure in the morning
Collect wild black soldier flies worm's ovum, withdraw the container being put in everywhere afternoon；
S3. by reclaim each container be placed on indoor cultivation, wait the wild worm's ovum for trapping cultivate to larva it is aging when, using Heisui River
Horsefly mature larva from food phase behavior black soldier flies mature larva is separated with formulation material；
S4. by the Morphological Identification of black soldier flies larva, black soldier flies mature larva is chosen；
S5. filter out that 20~25 days aging, every black soldier flies larva weighs 0.10~0.16g, the Heisui River that figure is plump, color is pitch-black
Horsefly mature larva, at ambient temperature, allow black soldier flies mature larva to pupate naturally, sprout wings after by obtain the black soldier flies of resistance to nicotine
Kind.
8. breeding method according to claim 6, it is characterised in that the side of the hot housing of the black soldier flies of the resistance to nicotine kind
Method is：
Starting stage：The ovum of the black soldier flies of resistance to nicotine kind is seeded in initial incubation material, the initial incubation material is by weighing as follows
The each group of amount percent is grouped into：Wheat bran 70~90%, imported fish meal 10~30%, with the nicotine of mass percent concentration 1 ‰
Above-mentioned initial incubation material is adjusted to humidity for 70% by aqueous solution；In room conditions, by above-mentioned ovum cultivate to larva it is aging, change
After pupa, emergence, adult is fed；
1st strain：After the Adult worms producting eggs of starting stage, the ovum for being produced is carried out using the 1st hot housing material
Hot housing；The 1st hot housing material is grouped into by each group of following percetage by weight：Wheat bran 70~90%, import fish
Powder 10~30%, it is 70% that above-mentioned hot housing material is adjusted to into humidity with the nicotine aqueous solution of mass percent concentration (1+S) ‰,
S is selected from 0.1~0.5；In room conditions, by above-mentioned ovum cultivate to larva it is aging, pupate, sprout wings after, feed adult；
The N+1 time strain：Hot housing, and N+1 are carried out again to the ovum that the adult obtained by n-th strain produces
Component contained by compost used by secondary strain meets following ratio：The mass percent concentration of nicotine aqueous solution=(n-th
Content+the S of nicotine in hot housing material) ‰, remaining constituent content is constant, and N is the natural number more than or equal to 1, repeats to strengthen, directly
To the mass percent concentration of nicotine aqueous solution in compost described in last time strain up to 3 ‰.
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| CN103283681A (en) * | 2012-03-05 | 2013-09-11 | 朱大恒 | Biological utilization method of waste tobacco stems |
| CN103563857B (en) * | 2013-11-07 | 2015-11-18 | 中国烟草总公司广东省公司 | A kind of screening technique of resistance to nicotine housefly kind |
| CN103563858B (en) * | 2013-11-07 | 2015-01-14 | 南雄市中大赛尔生物技术有限公司 | Reinforcing method for nicotine-resistant housefly species |
| CN103651263B (en) * | 2013-11-07 | 2015-12-09 | 中山大学 | A kind of breeding method of resistance to nicotine housefly strain |
| CN103975902B (en) * | 2014-05-30 | 2016-01-06 | 中山大学 | A kind ofly lure the method collecting wild black soldier flies |
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