CN107173337B - Method for preventing and treating vertical propagation of micro-particle disease embryo of silkworm double cross breeder seed - Google Patents
Method for preventing and treating vertical propagation of micro-particle disease embryo of silkworm double cross breeder seed Download PDFInfo
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- CN107173337B CN107173337B CN201710462792.8A CN201710462792A CN107173337B CN 107173337 B CN107173337 B CN 107173337B CN 201710462792 A CN201710462792 A CN 201710462792A CN 107173337 B CN107173337 B CN 107173337B
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- 241000255789 Bombyx mori Species 0.000 title claims abstract description 115
- 238000000034 method Methods 0.000 title claims abstract description 23
- 201000010099 disease Diseases 0.000 title claims abstract description 19
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title claims abstract description 19
- 210000001161 mammalian embryo Anatomy 0.000 title abstract description 11
- 239000011859 microparticle Substances 0.000 title abstract description 4
- 235000013601 eggs Nutrition 0.000 claims abstract description 75
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- 238000002791 soaking Methods 0.000 claims abstract description 17
- 208000015181 infectious disease Diseases 0.000 claims abstract description 15
- 238000001035 drying Methods 0.000 claims abstract description 12
- 238000005057 refrigeration Methods 0.000 claims abstract description 9
- 230000009965 odorless effect Effects 0.000 claims abstract description 5
- 238000007654 immersion Methods 0.000 claims abstract description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 20
- 230000000694 effects Effects 0.000 abstract description 14
- 230000002265 prevention Effects 0.000 abstract description 9
- 230000012447 hatching Effects 0.000 abstract description 8
- 238000011161 development Methods 0.000 abstract description 3
- 230000018109 developmental process Effects 0.000 abstract description 3
- 238000009402 cross-breeding Methods 0.000 abstract description 2
- 231100000989 no adverse effect Toxicity 0.000 abstract description 2
- 241000894007 species Species 0.000 abstract 1
- 241000243190 Microsporidia Species 0.000 description 6
- 238000011534 incubation Methods 0.000 description 6
- 230000001737 promoting effect Effects 0.000 description 6
- 238000007598 dipping method Methods 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 230000005484 gravity Effects 0.000 description 3
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 241001126828 Nosema bombycis Species 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- RPAJSBKBKSSMLJ-DFWYDOINSA-N (2s)-2-aminopentanedioic acid;hydrochloride Chemical class Cl.OC(=O)[C@@H](N)CCC(O)=O RPAJSBKBKSSMLJ-DFWYDOINSA-N 0.000 description 1
- 235000008708 Morus alba Nutrition 0.000 description 1
- 240000000249 Morus alba Species 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 241001126829 Nosema Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000005058 diapause Effects 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 230000013011 mating Effects 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 210000000287 oocyte Anatomy 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000009366 sericulture Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
- A01K67/04—Silkworms
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for preventing and treating infection of a micro-particle disease embryo of a silkworm double cross breeding species, which comprises the following steps: silkworm eggs without refrigeration treatment: after the silkworms lay eggs, standing the silkworm eggs for 15-24 h at 20-30 ℃, and then soaking the silkworm eggs in water at 47-48 ℃ for 6-10 min; taking out silkworm eggs, eluting with clear water until the silkworm eggs are odorless, and drying; silkworm eggs are refrigerated: taking the silkworm eggs out of a refrigeration house, standing for several hours at 15-20 ℃ as a buffer, and then putting the silkworm eggs into 47-50 ℃ water for immersion treatment for 6-10 min; taking out silkworm egg, eluting with clear water until no smell is produced, and drying. The relative prevention and treatment effect of the invention on the vertical propagation of the silkworm pebrine disease embryo seeds reaches 90-97%, the silkworm egg hatching rate can reach more than 92%, and no adverse effect is caused on the growth and development of the silkworm.
Description
Technical Field
The invention belongs to the field of silkworm production, and particularly relates to a method for preventing and treating vertical propagation of a pebrine disease embryo of a double cross stock of silkworms.
Background
Silkworm Nosema is an infectious silkworm disease which is caused by infecting silkworms with pathogenic microsporidia such as Nosema bombycis (Nosema bombycis). The disease can infect silkworms by eating mulberry leaves and also by infecting the silkworms with embryo seeds, so the disease is always valued by the industry and is the only object for silkworm quarantine.
The rapid development of silkworm in China in the eight and ninety years of the last century, and the laggard silkworm seed production facilities and the extensive technical management result in the national pandemic of silkworm diseases. In recent years, with the change of silkworm breeding areas, seed production environments and operation modes, the pebrine has the signs of death and reignition, and brings great economic loss to the sericulture industry.
At present, the varieties of domestic silkworms popularized and applied in the silkworm industry are all quaternary hybrid seeds, also called common seeds. It is a hybrid variety bred by using heterosis and adopting a mating mode of (A multiplied by B) multiplied by (C multiplied by D) for four silkworm varieties. The two varieties are first dually hybridized to produce double cross stock, and then the two double cross stocks are hybridized to produce quaternary hybrid. Therefore, the propagation of double cross stock is a major task in the stock farm of silkworms.
The prevention and treatment of the infection of the germ of the double cross breeding stock is an important guarantee for the production safety of the silkworm. However, silkworm pebrine lacks a specific treatment method, and the microsporidia is detected from the oocyte-producing moth under a microscope at present, and toxic silkworm seeds are removed according to a microscopic result, so that the infection of the embryo seeds of the silkworm pebrine is prevented. However, the method is related to the experience and technical level of virus testers, and is easy to cause misjudgment and missed judgment. Therefore, the search for new methods for preventing and treating the infection of the silkworm pebrine embryo is always the key direction of the scientific research of the silkworm industry.
Disclosure of Invention
Aiming at the current situation that the method for preventing and treating the infection of the embryo of the double cross bred silkworm is not complete, the invention aims to provide the method for preventing and treating the infection of the embryo of the micro particle disease of the double cross bred silkworm, and the aim of inactivating the microsporidian of the silkworm in the double cross bred silkworm is fulfilled by treating the double cross bred silkworm. The method is simple to operate, is practical and feasible, makes up the possible condition of missing judgment in the microscopic examination method, is a beneficial supplement to the microscopic examination method, and basically realizes the non-toxicity of the treated silkworm eggs.
The purpose of the invention is realized by the following technical scheme:
a method for preventing and treating the infection of the silkworm egg caused by the corpuscular disease of the double cross bred silkworm egg comprises the following steps:
silkworm eggs without refrigeration treatment: after the silkworms lay eggs, standing the silkworms at 20-30 ℃ for 15-24 h, and carrying out next treatment when the embryos of the silkworms develop to a germ band forming stage; then soaking the silkworm eggs in water at the temperature of 47-48 ℃ for 6-10 min, wherein the silkworm eggs are ensured to be uniformly and fully contacted with the solution during soaking; taking out silkworm eggs, eluting with clear water until the silkworm eggs are odorless, and drying;
silkworm eggs are refrigerated: taking the silkworm eggs out of a refrigeration house, standing for several hours at 10-20 ℃ as a buffer, and then putting the silkworm eggs into 47-50 ℃ water for dipping treatment for 6-10 min, wherein the silkworm eggs are ensured to be uniformly and fully contacted with the solution during dipping; taking out silkworm eggs, eluting with clear water until the silkworm eggs are odorless, and drying;
the silkworm eggs are double-cross stock;
said several hours are preferably 1 to 3 hours.
The water preferably contains acid or hydrogen peroxide;
the acid is hydrochloric acid, sulfuric acid, nitric acid or acetic acid, etc.;
the density of the water containing hydrochloric acid is 1.075-1.092g/cm3。
The method of the present invention depends on water temperature to kill microsporidian, but for silkworm double cross stock, the water temperature and soaking time need to be found and selected. When the water temperature is low, the microsporidia cannot be killed, and when the water temperature is too high, the hatching rate of the silkworm eggs is influenced; the same applies to the soaking time, which is short enough to kill microsporidia, and the soaking time is long enough to affect the hatching rate of the silkworm eggs. The invention pays creative labor for determining the two characteristics, obtains a proper range and obtains good effect. Meanwhile, the invention is dipped by dilute acid solution, which is beneficial to removing diapause of silkworm eggs and improving the hatching rate.
Compared with the prior art, the invention has the following advantages and effects:
1. the relative prevention and treatment effect of the invention on the vertical propagation of the microspherical disease embryo of the double cross stock silkworm reaches 90-97%.
2. The method of the invention can reach the silkworm egg hatching rate of more than 92 percent and has no adverse effect on the growth and development of the silkworms.
Drawings
FIG. 1 is a graph of the effect of immersion temperature on hatchability of inbreds that were not refrigerated.
FIG. 2 is the effect of immersion temperature on hatchability of cryopreserved diplodial stock.
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
A method for preventing and treating the infection of the silkworm egg caused by the corpuscular disease of the double cross bred silkworm egg comprises the following steps:
the two-way cross stock of 'Shuangguang II' is prepared by laying eggs, protecting the silkworm eggs at 25 deg.C for 20 hr, and placing at 47 deg.C with density of 1.075g/cm3Soaking in hydrochloric acid solution for 10min, eluting, drying, promoting incubation, and incubating. The practical hatchability reaches 94%, and the relative prevention and treatment effect reaches 95% compared with that of untreated contrast through microscopic examination of 3-year silkworms.
The calculation formula of the relative control effect is as follows:
relative control effect (%) (100-treatment group morbidity/control group morbidity) x 100
Example 2
A method for preventing and treating the infection of the silkworm egg caused by the corpuscular disease of the double cross bred silkworm egg comprises the following steps:
the two-way cross stock of the 'Shuangguang II' is used for protecting the silkworm eggs at 25 ℃ for 20 hours after the eggs are laid, and then the silkworm eggs are placed at 48 ℃ and the density is 1.075g/cm3Soaking in hydrochloric acid solution for 6min, eluting, drying, promoting incubation, and incubating. The practical hatchability reaches 93%, and the relative prevention and treatment effect reaches 96% compared with that of untreated control through microscopic examination of 3-year silkworms.
Example 3
A method for preventing and treating the infection of the silkworm egg caused by the corpuscular disease of the double cross bred silkworm egg comprises the following steps:
the two-way cross stock of the 'Shuangguang II' is used for protecting the silkworm eggs at 25 ℃ for 20 hours after laying eggs, and then the silkworm eggs are placed at 47.5 ℃ and the density is 1.075g/cm3Soaking in hydrochloric acid solution for 8min, eluting, drying, promoting incubation, and incubating. The practical hatchability reaches 94%, and the relative prevention and treatment effect reaches 97% compared with that of an untreated control through microscopic examination of 3-year silkworms.
Example 4
Under the same other operation conditions, experiments of different soaking temperature solutions were carried out on 9-Fule and 7-Xiang silkworm eggs of the double-cross stock, and the influence of the soaking temperature on the hatching rate of the silkworm eggs of the non-refrigerated double-cross stock is examined, and the result is shown in FIG. 1. It can be seen that the liquid temperature exceeds 48 ℃ and the silkworm egg hatching rate is significantly reduced, so that the upper limit of the temperature of the dipping solution is selected to be 48 ℃.
Example 5
A method for preventing and treating the infection of the silkworm egg caused by the corpuscular disease of the double cross bred silkworm egg comprises the following steps:
the two-way cross stock of 'Shuangguang No. two', is refrigerated and preserved, the silkworm eggs are taken out from a refrigeration house and protected for 2 hours at the intermediate temperature (15 ℃), and then placed at the temperature of 47 ℃ with the specific gravity of 1.092g/cm3Soaking in hydrochloric acid solution for 10min, eluting, drying, promoting incubation, and incubating. The practical hatchability reaches 94%, and the relative prevention and treatment effect reaches 92% compared with that of untreated contrast through microscopic examination of 3-year silkworms.
Example 6
A method for preventing and treating the infection of the silkworm egg caused by the corpuscular disease of the double cross bred silkworm egg comprises the following steps:
the two-way cross stock of 'Shuangguang No. two', is refrigerated and preserved, the silkworm eggs are taken out from a refrigeration house and protected for 2 hours at the intermediate temperature (15 ℃), and then the silkworm eggs are placed at the temperature of 50 ℃ and the specific gravity of the silkworm eggs is 1.092g/cm3Soaking in hydrochloric acid solution for 6min, eluting, drying, promoting incubation, and incubating. The practical hatchability reaches 93%, and the relative prevention and treatment effect reaches 93% compared with that of an untreated control through microscopic examination of 3-year-old silkworms.
Example 7
A method for preventing and treating the infection of the silkworm egg caused by the corpuscular disease of the double cross bred silkworm egg comprises the following steps:
the two-way cross stock of 'Shuangguang No. two', is refrigerated and preserved, the silkworm eggs are taken out from a refrigeration house and protected for 2 hours at the intermediate temperature (15 ℃), and then put into a refrigerator at the temperature of 48 ℃ and the specific gravity of 1.092g/cm3Soaking in hydrochloric acid solution for 8min, eluting, drying, promoting incubation, and incubating. The practical hatchability reaches 94%, and the relative prevention and treatment effect reaches 95% compared with that of untreated contrast through microscopic examination of 3-year silkworms.
Example 8
Under the same other operation conditions, experiments of different soaking temperature solutions were carried out on 9-Fule and 7-Xiang silkworm eggs of the double-cross stock, and the influence of the soaking temperature on the hatchability of the silkworm eggs of the refrigerated double-cross stock is examined, and the result is shown in FIG. 2. It can be seen that the liquid temperature exceeds 50 ℃ and the hatching rate of the silkworm eggs decreases significantly, so that the upper limit of the temperature of the dipping solution is selected to be 50 ℃.
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 (2)
1. A method for preventing and treating the infection of the silkworm egg caused by the corpuscular disease of the double cross bred silkworm egg is characterized by comprising the following steps:
silkworm eggs without refrigeration treatment: after the silkworms lay eggs, standing the silkworm eggs for 15-24 h at 20-30 ℃, and then soaking the silkworm eggs in water at 47-48 ℃ for 6-10 min; taking out silkworm eggs, eluting with clear water until the silkworm eggs are odorless, and drying;
silkworm eggs are refrigerated: taking the silkworm eggs out of a refrigeration house, standing for several hours at 10-20 ℃ as a buffer, and then putting the silkworm eggs into 47-50 ℃ water for immersion treatment for 6-10 min; taking out silkworm eggs, eluting with clear water until the silkworm eggs are odorless, and drying;
the water contains hydrochloric acid.
2. A method according to claim 1, characterized in that: the density of the water containing hydrochloric acid is 1.075-1.092g/cm3。
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| CN1687767A (en) * | 2005-05-11 | 2005-10-26 | 苏州大学 | Diseased eggs of nosemosis in product eggs of domestic silkworm, and method for detecting rate of diseased eggs |
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Application publication date: 20170919 Assignee: Guangdong Guangxin Seed Industry Co.,Ltd. Assignor: SERICULTURE & AGRI FOOD Research Institute GAAS Contract record no.: X2023980053696 Denomination of invention: A Prevention and Control Method for Vertical Transmission of Microparticle Disease in Embryos of Double Crossed Original Silkworm Granted publication date: 20200414 License type: Common License Record date: 20231222 |
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