CN109757444B - Method for treating silkworm pebrine by targeted heat treatment of silkworm seeds - Google Patents
Method for treating silkworm pebrine by targeted heat treatment of silkworm seeds Download PDFInfo
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- CN109757444B CN109757444B CN201910225294.0A CN201910225294A CN109757444B CN 109757444 B CN109757444 B CN 109757444B CN 201910225294 A CN201910225294 A CN 201910225294A CN 109757444 B CN109757444 B CN 109757444B
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Abstract
A method for treating silkworm pebrine by targeted heat treatment of silkworm eggs comprises the following steps: soaking silkworm eggs in acid, continuously carrying out two times of heat treatment at intervals of 24-48h, and finally carrying out incubation until silkworm eggs are hatched; and (3) carrying out heat treatment on silkworm eggs with the egg age of 4-28 h for refrigerating silkworm eggs, carrying out heat treatment again after 24-48h, then warehousing and refrigerating, pickling after the silkworm eggs are taken out of a warehouse, and accelerating hatching. Compared with a high-temperature pickling method, the method disclosed by the invention can be used for thoroughly inactivating the nosema bombycis in the silkworm seeds, ensuring the safety of the silkworm seeds and meeting the conditions for popularization and application in production. After the method disclosed by the invention is disclosed, the silkworm quarantine link is hopefully cancelled, so that the silkworm production efficiency and economic benefit are obviously improved. Compared with chemical medicines, the invention is physical treatment, is environment-friendly and safe, and has no toxic or side effect on silkworm eggs.
Description
Technical Field
The invention belongs to the field of silkworm disease control, and particularly relates to a method for treating silkworm pebrine by targeted heat treatment of silkworm eggs.
Background
Nosema bombycis disease caused by silkworm microsporidia (Nosema bombycis) infecting silkworm is a destructive epidemic disease in silkworm production. The disease can infect silkworms by eating mulberry leaves and can also infect filial generation through eggs, so that the silkworm breeding is lost or even the silkworm does not accept granules, and the disease is always valued by the industry and is listed as the only legal quarantine object by all silkworm producing countries.
In recent years, with the change of silkworm breeding environment, seed production environment and operation mode, the occurrence situation of the pebrine disease in China becomes more severe, and the quantity of the silkworm seeds eliminated due to the ultra-toxicity is high, so that the economic loss caused each year reaches thousands of yuan.
At present, there is no specific and effective treatment means for the pebrine disease. The silkworm breeding farm is mainly characterized in that a silkworm breeding place and tools are disinfected by a disinfectant for silkworms, and pathogens in the environment are purified, so that infection of the silkworms is avoided. "carbendazim powder for silkworm" produced by Guangdong four seasons mulberry field silkworm science and technology limited company is the only silkworm microparticle disease treatment drug which obtains production batch, but the drug can generate toxic and side effects on silkworms after long-term use, and the yield and quality of silkworm eggs are influenced.
The Chinese patent ZL 201710462723.7 discloses a method for preventing and treating the vertical propagation of embryo seeds of the pebrine silkworm pebrine disease of quaternary hybrid silkworm, which can effectively block the vertical propagation of the embryo seeds of the pebrine silkworm disease by carrying out high-temperature pickling treatment on the virus-carrying silkworm seeds with the egg age of 15-30 h. In the period, the pathogeny in the disease egg is guessed to be mainly in a schizont state, and the heat treatment can kill the schizont without adverse effect on the silkworm embryo.
However, the method of the above patent suffers from two significant drawbacks:
(1) mature spores in sick eggs cannot be inactivated, so that the spores continue to grow and proliferate in silkworm eggs and larva bodies, and the safety of silkworm eggs cannot be guaranteed;
(2) compared with qualified silkworm, the life rate and the cocooning rate of the silkworm larvae are reduced, and the treatment effect is unstable.
Therefore, the method of the above patent faces great difficulty in practical production.
Disclosure of Invention
Aiming at the defect that mature spores in diseased eggs cannot be inactivated by the existing method, the invention aims to provide a method for treating the nosema bombycis disease by targeted heat treatment of silkworm eggs. The method has strong operability and lower cost, the treated silkworm eggs can be normally incubated to complete the development of the whole generation, and the method has no toxic or side effect on offspring and does not influence the economic character of the offspring.
The purpose of the invention is realized by the following technical scheme:
a method for treating silkworm pebrine by targeted heat treatment of silkworm seeds comprises the following steps:
namely soaking silkworm seeds: pickling silkworm eggs with the egg age of 14-28 h, carrying out incubation for 24-48h, and carrying out hot air treatment or hot water bath treatment; continuously carrying out incubation for 24-48h, and carrying out hot air treatment or hot water bath treatment again (the development of silkworm eggs can be influenced by continuous heat treatment); finally, hastening hatching until the silkworm eggs are hatched;
the pathogeny in the silkworm eggs at the initial stage of spawning is mainly schizont and can be inactivated by high temperature, so that the schizont can be killed by soaking treatment within 20h, but the mature spores can not be killed, which is an imperfect place in the prior patent, because the undecided mature spores can continue to grow and proliferate.
According to the proliferation rule (figure 1, embodiment 1) of the pathogen in the silkworm eggs after the poisonous silkworm eggs are immediately pickled, that the pathogen grows at the proliferation peak after 24 hours and 48 hours after soaking, the proliferation stage that mature spores can grow to schizonts after 24-48 hours is an effective time period for continuous heat treatment, and the method is called targeted heat treatment. Like tumor chemotherapy, cancer cells in vivo can be completely killed by several successive chemotherapies. Similarly, through several times of targeted heat treatment, the pathogens in the silkworm eggs can be thoroughly killed, so that a better treatment effect is obtained.
Refrigerating silkworm eggs: carrying out hot air treatment or hot water bath treatment on silkworm eggs with the egg age of 4-28 h, protecting for 24-48h at 25 ℃, carrying out hot air treatment or hot water bath treatment again, and then selecting the silkworm eggs according to the egg-using time to put in a warehouse for refrigeration; pickling silkworm eggs after leaving the house, and hatching;
as above, the initial stage of spawning is the schizont stage, which is the effective target period for heat treatment. However, since the silkworm eggs are refrigerated (i.e. the refrigerated silkworm eggs cannot be hatched immediately, but are refrigerated for a certain period of time as required and then taken out of the house), the silkworm eggs cannot be treated by pickling (since pickling is equivalent to preventing diapause of the silkworm eggs, the silkworm eggs will be hatched), and other heat treatment measures are only available.
According to the invention, the proliferation rule of pathogens in the silkworm eggs after the eggs of the poisonous silkworm eggs are laid (figure 2 and example 3) shows that the pathogen amount in the silkworm eggs at the 2 nd day is increased sharply, the proliferation peak period of the nosema bombycis at 1-2 d after the eggs are laid is presumed, and the pathogen amount per day has no significant change and is probably related to that the silkworm eggs gradually enter a diapause state and the growth and development of the pathogens are also inhibited.
The silkworm seeds are all silkworm seeds of original stock, stock and common stock.
The pickling conditions are as follows: the acid solution has a density of 1.075-1.092g/cm3The temperature of the acid solution is 46-48 ℃,pickling for 5-12 min;
the silkworm egg hatching is carried out at the ambient temperature of 27 +/-1 ℃ and the humidity of 80-85%.
The hot air treatment is carried out at the temperature of 46-50 ℃ for 20-80 min;
and performing hot water bath treatment at the temperature of 46-50 ℃ for 1-12 min.
The method of the invention is based on targeted heat treatment of silkworm eggs with poison, thereby completely inactivating the nosema bombycis in silkworm eggs. The invention carries out systematic research on the growth and proliferation rules of pathogens in the silkworm eggs during the period from egg laying to hatching, realizes the targeted positioning of pathogen schizonts on the basis, provides a specific method for targeted heat treatment of silkworm eggs, and optimizes specific conditions of heat treatment by taking the silkworm egg hatching rate as an index. The present invention is creative in this regard, and achieves a remarkable therapeutic effect compared to the previous methods.
Compared with the prior art, the invention has the following advantages and effects:
1. compared with chemical medicines, the invention is physical treatment, is environment-friendly and safe, and has no toxic or side effect on silkworm eggs.
2. Compared with a high-temperature pickling method, the method disclosed by the invention can be used for thoroughly inactivating the nosema bombycis in the silkworm seeds, ensuring the safety of the silkworm seeds and meeting the conditions for popularization and application in production.
3. After the method disclosed by the invention is disclosed, the silkworm quarantine link is hopefully cancelled, so that the silkworm production efficiency and economic benefit are obviously improved.
Drawings
FIG. 1 shows the proliferation rule of pathogen in silkworm eggs after the instant pickling of the virus-carrying silkworm eggs.
FIG. 2 shows the proliferation rule of pathogen in silkworm eggs after the eggs of the silkworm with poison are laid.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
Example 1
Eluting the 'two Guang No. two' silkworm eggs with toxicity into loose eggs, carrying out instant pickling treatment on the silkworm eggs at the egg age of 20h (47 ℃, 7min), then putting the silkworm eggs into a hatching chamber for hatching, sampling 40 silkworm eggs every 24h, extracting DNA, carrying out absolute quantification on pathogen in the silkworm eggs through fluorescent quantitative PCR reaction, and analyzing the proliferation rule of the pathogen in the silkworm eggs after the instant pickling.
As a result, referring to FIG. 1, it can be seen that the pathogens showed proliferation peaks at 24h (2 d after egg laying) and 48h (3 d after egg laying) after the immersion, which is the preferable time for the heat treatment.
It is to be explained that in FIG. 1, the number of schizonts is highest at 8d, but it can be seen that 24-48h is the first peak, where the schizonts are absolutely dominant and decrease by 4-6d, at which stage the schizonts have developed into mature spores. Then division and proliferation are carried out again in 7-8 days, so it is high.
The selection of 24-48h for heat treatment is also based on the following general considerations:
(1) attack pathogeny, certainly when the amount of the pathogeny is small, but not when the amount of the pathogeny in the body is large;
(2) by 8 days, the silkworm egg embryo is rapidly developed into a newly-hatched silkworm, the tolerance is reduced, and the heat treatment affects the silkworm embryo.
Example 2
A method for treating silkworm pebrine by targeted heat treatment of silkworm seeds comprises the following steps:
the method comprises the following steps of carrying out instant pickling treatment on silkworm eggs 20 hours after spawning on the 'two Guangdong No. two' quaternary hybrid seeds (47 ℃, 7min), then putting the silkworm eggs into a hatching chamber for hatching, carrying out targeted hot air treatment (46 ℃, 60min) or hot water bath treatment (47 ℃, 6min) after 24 hours, continuing hatching, carrying out targeted hot air treatment or hot water bath treatment again after 24 hours, and continuing hatching until the silkworm eggs are hatched. The practical hatchability reaches 93 percent, and the incidence rate is only 2.58 percent through microscopic examination of 3-year-old silkworms.
Example 3
Eluting the 'two Guang No. two' silkworm eggs with toxin into loose eggs, standing at 25 ℃, sampling 40 silkworm eggs every 24 hours, extracting DNA, absolutely quantifying pathogen in the silkworm eggs through fluorescence quantitative PCR reaction, and analyzing the proliferation rule of the pathogen in the silkworm eggs after egg laying.
As a result, as shown in FIG. 2, it can be seen that the pathogen amount in the eggs at 2d shows a sharp increase, which indicates that 1-2 d after egg laying is the proliferation peak period of nosema bombycis, which is the preferred time for heat treatment.
Example 4
A method for treating silkworm pebrine by targeted heat treatment of silkworm seeds comprises the following steps:
and (3) carrying out hot air treatment or hot water bath treatment on silkworm eggs with the egg age of 20h, protecting at 25 ℃ for 24h, carrying out hot air treatment or hot water bath treatment again, and warehousing and refrigerating the silkworm eggs when the silkworm eggs are 72 h. And (5) pickling after delivery, and accelerating incubation and hatching. The practical hatchability reaches 95 percent, and the incidence rate is only 1.97 percent when the 3-year-old silkworms are examined by a microscope.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (4)
1. A method for treating silkworm pebrine by targeted heat treatment of silkworm seeds is characterized by comprising the following steps:
namely soaking silkworm seeds: pickling silkworm eggs of 20h of egg age, carrying out incubation for 24h, and carrying out hot air treatment; continuously carrying out green-keeping for 24h, and carrying out hot air treatment again; finally, hastening hatching until the silkworm eggs are hatched;
refrigerating silkworm eggs: carrying out hot air treatment on silkworm eggs with the egg age of 20h, protecting the silkworm eggs at 25 ℃ for 24h, carrying out hot air treatment again, and then putting the silkworm eggs into a warehouse for refrigeration according to the seed using time; pickling silkworm eggs after leaving the house, and hatching;
the hot air treatment is carried out at the temperature of 46-50 ℃ for 20-80 min;
the silkworm eggs are of two Guangdong provinces and two types.
2. The method of claim 1, wherein: the acid solution is 1.075-1.092g/cm3Hydrochloric acid of (2).
3. The method of claim 1, wherein: and pickling, wherein the temperature of the acid liquor is 46-48 ℃, and the pickling time is 5-12 min.
4. The method of claim 1, wherein: the silkworm egg hatching is carried out at the ambient temperature of 27 +/-1 ℃ and the humidity of 80-85%.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107041342A (en) * | 2017-06-19 | 2017-08-15 | 广东省农业科学院蚕业与农产品加工研究所 | For the prevention and controls of tetra hybrid pebrine disease semina vertical transmission |
| CN107173337A (en) * | 2017-06-19 | 2017-09-19 | 广东省农业科学院蚕业与农产品加工研究所 | A kind of prevention and controls for silkworm double cross original seed granulosis semina vertical transmission |
| CN107232149A (en) * | 2017-06-19 | 2017-10-10 | 广东省农业科学院蚕业与农产品加工研究所 | A kind of prevention and controls for silkworm original silkworm egg granulosis semina vertical transmission |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107041342A (en) * | 2017-06-19 | 2017-08-15 | 广东省农业科学院蚕业与农产品加工研究所 | For the prevention and controls of tetra hybrid pebrine disease semina vertical transmission |
| CN107173337A (en) * | 2017-06-19 | 2017-09-19 | 广东省农业科学院蚕业与农产品加工研究所 | A kind of prevention and controls for silkworm double cross original seed granulosis semina vertical transmission |
| CN107232149A (en) * | 2017-06-19 | 2017-10-10 | 广东省农业科学院蚕业与农产品加工研究所 | A kind of prevention and controls for silkworm original silkworm egg granulosis semina vertical transmission |
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