CN107114561B - Induced mulberry leaf for preventing and treating bombyx mori BmNPV disease and application thereof - Google Patents

Induced mulberry leaf for preventing and treating bombyx mori BmNPV disease and application thereof Download PDF

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CN107114561B
CN107114561B CN201710252959.8A CN201710252959A CN107114561B CN 107114561 B CN107114561 B CN 107114561B CN 201710252959 A CN201710252959 A CN 201710252959A CN 107114561 B CN107114561 B CN 107114561B
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mulberry leaves
bmnpv
induced
ultraviolet light
silkworm
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CN107114561A (en
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朱玮
田景奎
杨丙贤
张琳
胡瑾
虞姣姣
管祺杰
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Kaihua Mingyang Health Technology Co.,Ltd.
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Zhejiang University ZJU
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/90—Feeding-stuffs specially adapted for particular animals for insects, e.g. bees or silkworms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/60—Moraceae (Mulberry family), e.g. breadfruit or fig
    • A61K36/605—Morus (mulberry)

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
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  • Food Science & Technology (AREA)
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  • Animal Husbandry (AREA)
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  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
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  • Pharmacology & Pharmacy (AREA)
  • Molecular Biology (AREA)
  • Physiology (AREA)
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  • Alternative & Traditional Medicine (AREA)
  • Medical Informatics (AREA)
  • Insects & Arthropods (AREA)
  • Birds (AREA)
  • Fodder In General (AREA)
  • Feed For Specific Animals (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

The invention discloses an induced mulberry leaf for preventing and treating bombyx mori BmNPV disease and application thereof. Silkworm nuclear polyhedrosis virus is one of the viruses causing the greatest harm to silkworms, and no clear treatment method for BmNPV disease exists at present. The method comprises the steps of irradiating fresh mulberry leaves by ultraviolet light, wherein the ultraviolet light is one or a combination of more of UVA wave band, UVB wave band and UVC wave band, the irradiation intensity of the ultraviolet light is 10-1000W, the ultraviolet irradiation time is 0.05-3h, and the humidity is kept at 10% -100% in the irradiation process. The mulberry leaves have important pharmacological activity and are the main food sources of the silkworms. The invention can obviously change the content of secondary metabolites of mulberry leaves, reduce the death rate of the silkworm infected with virus and resist the silkworm nuclear polyhedrosis virus.

Description

Induced mulberry leaf for preventing and treating bombyx mori BmNPV disease and application thereof
Technical Field
The invention belongs to insect breeding, and particularly relates to induced mulberry leaves for preventing and treating bombyx mori BmNPV disease and application thereof.
Background
Silkworm (Bombyx mori), Lepidoptera, Bombycidae, and silkworm cocoon, and is eaten by mulberry leaves. The bombyx mori nuclear polyhedrosis virus (BmNPV) particle is rod-shaped, belongs to the family of baculovirus, the genus of baculovirus, and the species of bombyx mori nuclear polyhedrosis virus, which is a representative species of single capsid nuclear polyhedrosis virus. The outer envelope membrane contains double-stranded DNA molecules, is one of the most sensitive and common pathogens of silkworm species, and is festered and killed after the infection of silkworms, and the reaction is severe and rapid, so that huge economic loss is caused in the silkworm industry. After the virus infects silkworm, ODV is released in alkaline midgut fluid, and then the ODV passes through a feeding membrane and infects midgut columnar epithelial cells, the virus removes the cyst membrane, the nucleocapsid enters into cell nucleus and the virus is replicated and assembled to form new virus particles and polyhedra. The replicated nucleocapsid is released early in the infection to form BV. In the late stages of infection, polyhedrin encapsulates the newly formed cyst membrane protein-coated ODV to form an OB. During infection, virions use the replicated DNA and synthetic proteins for tissue assembly, acquire envelopes from host cell membranes through budding, after which viral infection spreads throughout the host and can cause secondary infections using the bombyx mori tracheal system, resulting in host disruption. After viral infection, the body of silkworms swell, fester and die, after which OB is released into the environment to restart a new round of infection. The resistance of the silkworm to BmNPV can also be influenced by the growth environment, the quality of mulberry leaves and the like, for example, the proper temperature and humidity are beneficial to the growth of the silkworm, so that the resistance of the silkworm can be enhanced, and the effective and safe disinfectant can prevent virus infection, for example, the disinfectant liquid mixed by dibromohydantoin and sodium carbonate has the disinfection effect on pathogens. However, there is currently no clear way to treat BmNPV disease.
Mulberry (Morus alba L.) is a plant of Morus genus of Moraceae family. Mulberry leaf, being cold in nature and sweet and bitter in taste, has the functions of dispelling wind and heat, clearing lung-heat and moistening dryness, and clearing liver and improving vision, and is one of the commonly used traditional Chinese medicines. The mulberry leaves contain more secondary metabolites, have the functions of tumor resistance, oxidation resistance, virus resistance and the like, and are important medicinal plants. After folium mori responds to ultraviolet induction, secondary metabolites are significantly changed.
Disclosure of Invention
Aiming at the defects of the prior art and the current situation of prevention and treatment of the BmNPV disease of the silkworms, the invention provides an induced mulberry leaf for preventing and treating the BmNPV disease of the silkworms and application thereof. The induced mulberry leaf can obviously reduce the death rate of the silkworm infected with BmNPV. The invention adopts the following specific technical scheme:
induced mulberry leaves for preventing and treating bombyx mori BmNPV disease are obtained after the mulberry leaves are irradiated by ultraviolet light; the ultraviolet light is one or a combination of more of UVA wave band, UVB wave band and UVC wave band. The mulberry leaves have important pharmacological activity and are the main food sources of the silkworms. After the ultraviolet irradiation, the content of secondary metabolites of mulberry leaves can be obviously changed, the death rate of the silkworm infected with virus is reduced, and the silkworm nuclear polyhedrosis virus is resisted.
Based on this technical solution, a plurality of preferred embodiments can be provided, and technical features in the preferred embodiments can be combined with each other without conflicting with each other, without limitation.
Preferably, the mulberry leaves are picked fresh mulberry leaves.
Preferably, the picking time of the mulberry leaves is 4-10 months.
Preferably, the ultraviolet irradiation intensity is 10 to 1000W.
Preferably, the air humidity is maintained between 10% and 100% during irradiation.
Preferably, the irradiation time of the mulberry leaves by the ultraviolet light is 0.05 to 3 hours.
The induced mulberry leaves prepared by any scheme can be used for preparing corresponding feeds and can also be used for preparing medicines for preventing and treating the BmNPV diseases of the silkworms.
The invention can obviously change the content of secondary metabolites of mulberry leaves, and the death rate of the silkworms infected with BmNPV can be obviously reduced after the silkworms are fed with the feed, so that the silkworm nuclear polyhedrosis virus can be resisted.
Detailed Description
The present invention is further illustrated by the following specific examples so that those skilled in the art can better understand the essence of the present invention.
Example 1
The mulberry leaves which are freshly picked in 6 months are irradiated for 10 minutes by UVB ultraviolet light with the intensity of 1000W, the air humidity is kept at 40 percent in the irradiation process, and the obtained mulberry leaves are used as induced mulberry leaves. Silkworm is divided into two groups, one group is fed with normal fresh mulberry leaves, the other group is fed with induced mulberry leaves, each group contains 90 silkworm leaves, the first day of the second age is inoculated with 200 muL BmNPV, and the mulberry leaves are sprayed with 5 diseases with different concentrationsToxic, i.e. 1.22 × 104units/mL，1.22×105units/mL，1.22×106units/mL，1.22×107units/mL，1.22×108In this experiment, water was used instead of the virus as a control group, and as a result, it was found that the half lethal concentration (LC50) of BmNPV of the silkworm group bred with normal fresh mulberry leaves was (1.86. + -. 0.15) × 106LC50 of BmNPV of the silkworm group bred to induce mulberry leaves was (3.30. + -. 0.51) × 106unit/mL, the induction of mulberry leaves can obviously reduce the death rate of silkworms after virus infection.
Example 2
Fresh mulberry leaves picked in 7 months are irradiated by UVA ultraviolet light with the intensity of 200W for 30 minutes, the air humidity is kept at 50% in the irradiation process, the obtained mulberry leaves are used as induced mulberry leaves, silkworms are divided into two groups, one group is fed with normal fresh mulberry leaves, the other group is fed with induced mulberry leaves, each group contains 90 mulberry leaves, inoculation is carried out on the first day of the second age by 200 mu L BmNPV, 5 viruses with different concentrations are sprayed on the mulberry leaves, namely 1.22 × 104units/mL，1.22×105units/mL，1.22×106units/mL，1.22×107units/mL，1.22×108In this experiment, water was used instead of the virus as a control group, and as a result, LC50 of BmNPV of a silkworm group bred with normal fresh mulberry leaves was found to be (1.83. + -. 0.09) × 106LC50 of BmNPV of the silkworm group bred to induce mulberry leaves was (2.51. + -. 0.43) × 106unit/mL, the induction of mulberry leaves can obviously reduce the death rate of silkworms after virus infection.
Example 3
The method comprises the steps of picking fresh mulberry leaves in 8 months, keeping the air humidity at 60% in the irradiation process for 2 hours under UVB ultraviolet light with the intensity of 20W, taking the obtained mulberry leaves as induced mulberry leaves, dividing silkworms into two groups, feeding normal fresh mulberry leaves in one group, feeding induced mulberry leaves in one group, inoculating 90 mulberry leaves in 200 mu L BmNPV in the first day of the third year, and spraying 5 viruses with different concentrations on the mulberry leaves, namely 1.22 × 104units/mL，1.22×105units/mL，1.22×106units/mL，1.22×107units/mL，1.22×108units/mL. In this experiment, water was usedAs a result, it was found that the half lethal concentration (LC50) of BmNPV in the silkworm group bred with normal fresh mulberry leaves was (1.94. + -. 0.35) × 106LC50 of BmNPV of the silkworm group bred to induce mulberry leaves was (2.87. + -. 0.21) × 106unit/mL, the induction of mulberry leaves can obviously reduce the death rate of silkworms after virus infection.
Example 4
The method comprises the steps of picking fresh mulberry leaves in 9 months, irradiating the fresh mulberry leaves for 60 minutes under UVC ultraviolet light with the intensity of 400W, keeping the air humidity at 50% in the irradiation process, taking the obtained mulberry leaves as induced mulberry leaves, dividing silkworms into two groups, feeding normal fresh mulberry leaves in one group, feeding induced mulberry leaves in one group, inoculating 90 mulberry leaves in each group at 200 mu L BmNPV in the first day of the third year of age, and spraying 5 viruses with different concentrations on the mulberry leaves, namely 1.22 × 104units/mL，1.22×105units/mL，1.22×106units/mL，1.22×107units/mL，1.22×108In this experiment, water was used instead of the virus as a control group, and as a result, it was found that the half lethal concentration (LC50) of BmNPV of the silkworm group bred with normal fresh mulberry leaves was (1.95. + -. 0.25) × 106LC50 of BmNPV of the silkworm group bred to induce mulberry leaves was (3.17. + -. 0.37) × 106unit/mL, the induction of mulberry leaves can obviously reduce the death rate of silkworms after virus infection.
The above-described embodiments are merely preferred embodiments of the present invention, which should not be construed as limiting the invention. Various changes and modifications may be made by one of ordinary skill in the pertinent art without departing from the spirit and scope of the present invention. For example, the ultraviolet irradiation intensity, the air humidity and the irradiation time can be adjusted according to the test, the ultraviolet irradiation intensity is 10-1000W, the air humidity is 10-100%, and the ultraviolet irradiation time is 0.05-3 hours. Therefore, the technical scheme obtained by adopting the mode of equivalent replacement or equivalent transformation is within the protection scope of the invention.

Claims (1)

1. An application of induced mulberry leaves in preparing a medicament for preventing and treating bombyx mori BmNPV diseases is disclosed, wherein the preparation method of the induced mulberry leaves comprises the following steps: irradiating mulberry leaves by ultraviolet light to obtain induced mulberry leaves; the ultraviolet light is one or a combination of more of UVA wave band, UVB wave band and UVC wave band; the mulberry leaves are picked fresh mulberry leaves, and the picking time of the mulberry leaves is 4-10 months; the ultraviolet light irradiation intensity is 10-1000W, the air humidity is kept at 10-100% in the irradiation process, and the irradiation time of the ultraviolet light on the mulberry leaves is 0.05-3 hours.
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CN108034641A (en) * 2018-01-24 2018-05-15 浙江大学 Isolation and purification method of Diels-Alderase enzymes and application thereof in a kind of uv induction mulberry leaf
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CN103690534A (en) * 2013-12-04 2014-04-02 西南大学 Application of prodiginine (PG) in prevention and control of bombyx morinuclear polyhedrosis virus
CN103947614A (en) * 2014-04-24 2014-07-30 苏州大学 Method for inhibiting in-vivo BmNPV viral multiplication of silkworms

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CN101875647A (en) * 2010-05-31 2010-11-03 杭州柏希特医药科技有限公司 Method for improving plant antibiotic chalcomoracin content in mulberry leaves

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CN103947614A (en) * 2014-04-24 2014-07-30 苏州大学 Method for inhibiting in-vivo BmNPV viral multiplication of silkworms

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