CN105456285A - Method for reducing bacterial infection during production of biological agent by adopting silkworms or silkworm chrysalis - Google Patents
Method for reducing bacterial infection during production of biological agent by adopting silkworms or silkworm chrysalis Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
- A61K31/7056—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing five-membered rings with nitrogen as a ring hetero atom
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/425—Thiazoles
- A61K31/427—Thiazoles not condensed and containing further heterocyclic rings
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
- A61K31/542—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame ortho- or peri-condensed with heterocyclic ring systems
- A61K31/545—Compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins, cefaclor, or cephalexine
- A61K31/546—Compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins, cefaclor, or cephalexine containing further heterocyclic rings, e.g. cephalothin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/7036—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin having at least one amino group directly attached to the carbocyclic ring, e.g. streptomycin, gentamycin, amikacin, validamycin, fortimicins
Abstract
The invention discloses a method for reducing bacterial infection during the production of a biological agent by adopting silkworms or silkworm chrysalis. According to the method, when the biological agent is produced by utilizing a genetically modified baculovirus expression system and by adopting silkworms or silkworm chrysalis as a host carrier, or when silkworm insect grass products or cordyceps militaris products are produced by utilizing the silkworms or silkworm chrysalis as a host carrier, 2-3 kinds of different antibiotics are matched and mixed to be used for the host, so that bacterial diseases are inhibited, wherein the silkworms are bombyx or tussah, the silkworm chrysalis is bombyx puparium or tussah puparium, and the use method of antibiotics adopts puncture injection. According to the invention, the existing antibiotic mixing technology is adopted, the effective antibiotics are selected and mixed at an appropriate ratio, so that the bacterial infection is reduced, the ratio of the healthy host silkworms or silkworm chrysalis is effectively improved, the death rate of the silkworms or silkworm chrysalis is greatly reduced, the expression efficiency of exogenous genes taking silkworms or silkworm chrysalis as the expression carrier is improved, the production efficiency is improved, and the production cost is reduced. The method is simple and reliable, and is suitable for large-scale production application.
Description
Technical field
The present invention relates to biological technology application, specifically a kind of method reducing bacteriological infection when adopting silkworm or Pupa bombycis to produce biological preparation.
Background technology
At present with silkworm or Pupa bombycis for expressive host, the mode of producing biological preparation mainly contains two kinds, one is the production model adopting polyhedrosis recombinant baculovirus to carry out massive duplication production exogenous gene in host, as the M protein gene, foot-and-mouth disease antigen combination gene, insulin like growth factor II etc. of hepatitis B surface antigen: another kind is for host produces Cordyceps militaris bacterium, as silkworm grass and Cordyceps militaris (L.) Link. with silkworm or pupa.But in specific implementation process often because of operating environment or vaccinal injection utensil unclean and infected the silkworm of pathogenic bacteria or Pupa bombycis and to be caused by the secondary cross infection of Inoculating needle and by the silkworm or Pupa bombycis being connect profit, bacterial disease is occurred in a large number and infect and form deteriorated blood silkworm or deteriorated blood pupa makes production failure.
Summary of the invention
The method of bacteriological infection is reduced, to solve the problem proposed in above-mentioned background technology when the object of the present invention is to provide a kind of simple and reliable, employing silkworm that production cost is low or Pupa bombycis to produce biological preparation.
For achieving the above object, the invention provides following technical scheme:
A kind of method reducing bacteriological infection when adopting silkworm or Pupa bombycis to produce biological preparation, be adopt utilize transgenic baculovirus expression system to produce biological preparation using silkworm or Pupa bombycis as host cell or utilize silkworm or Pupa bombycis to produce silkworm grass or Cordyceps militaris (L.) Link. product as host cell time, by different types of for 2-3 kind antibiosis rope interworking conbined usage in host, bacterial disease is suppressed.
As the further scheme of the present invention: described silkworm is silkworm or Antherea pernyi Guerin-Meneville, Pupa bombycis is silkworm pupa or Antheraea pernyi Geurin Meneville.
As the further scheme of the present invention: described transgenic baculovirus is bombyx mori nuclear polyhydrosis virus BmNPV and Antheraea pernyi nuclear polyhedrosis virus ApNPV, described transgenic baculovirus expression system refers to be cultivated in specific recombinant baculovirus or gene insertion silkworm or Pupa bombycis body, can the recombinant baculovirus implanted or gene be transcribed and be translated, the bioactive substance that Nature creating is useful to the mankind in silkworm or Pupa bombycis body.
As the further scheme of the present invention: described silkworm grass and Cordyceps militaris (L.) Link. refer to that the domestication of Chinese caterpillar fungus strain spore is inoculated into silkworm body or Pupa bombycis body, carrying out parasitic life with the material in silkworm body or Pupa bombycis body, finally to stretch out silkworm body or Pupa bombycis body surface after Sum decomposition and organism that a kind of fungus of being formed and silkworm body or Pupa bombycis body case coexist by growing.
As the further scheme of the present invention: described antibiotic refers to the aztreonam of beta-lactam, cefoperazone sodium, lincomycin class lincomycin hydrochloride, aminoglycoside streptomycin or fluoroquinolone antibiotics ciprofloxacin.
As the further scheme of the present invention: the using method of described antibiosis rope is puncture injection.
As the further scheme of the present invention: described antibiotic service time is five length of times and pupa time of silkworm.
As the further scheme of the present invention: described antibiotic use amount is as follows: with silkworm or silkworm pupa for the consumption of every bar silkworm or every pupa during host is: aztreonam is 40 ~ 80 μ g, cefoperazone sodium is 60 ~ 100 μ g, lincomycin hydrochloride is 15 ~ 40 μ g, streptomycin is 5 ~ 15 μ g, ciprofloxacin 4 ~ 8 μ g; With Antherea pernyi Guerin-Meneville or Antheraea pernyi Geurin Meneville for the consumption of every bar silkworm or every pupa during host is: aztreonam is 80 ~ 160 μ g, cefoperazone sodium is 120 ~ 200 μ g, lincomycin hydrochloride is 35 ~ 65 μ g, streptomycin is 10 ~ 20 μ g, salt ciprofloxacin 8 ~ 16 μ g.
As the present invention's further scheme: described antibiotic before use, by above-mentioned dosage and transgenic baculovirus liquid or silkworm Cordyceps liquid or Cordyceps militaris (L.) Link. fungus liquid according to injecting object dosage mixture.
Compared with prior art, the invention has the beneficial effects as follows:
The present invention adopts existing antibiotic interworking technology, effective antibiosis rope is selected to mix with appropriate ratio, the ratio that bacteriological infection effectively improves health hosts silkworm or Pupa bombycis can be reduced, significantly reduce silkworm or Pupa bombycis mortality rate, improving exogenous gene producing the production efficiency of silkworm grass or Cordyceps militaris (L.) Link. using silkworm or Pupa bombycis as carrier as expression efficiency during expression vector and raising using silkworm or Pupa bombycis, reducing production cost; The method is simple and reliable, is applicable to large-scale production application.
Detailed description of the invention
Be described in more detail below in conjunction with the technical scheme of detailed description of the invention to this patent.
A kind of method reducing bacteriological infection when adopting silkworm or Pupa bombycis to produce biological preparation, be adopt utilize transgenic baculovirus expression system to produce biological preparation using silkworm or Pupa bombycis as host cell or utilize silkworm or Pupa bombycis to produce silkworm grass or Cordyceps militaris (L.) Link. product as host cell time, by different types of for 2-3 kind antibiotic interworking conbined usage in host, bacterial disease is suppressed, the ratio of effective raising health hosts silkworm or Pupa bombycis, increases substantially production efficiency and reduces production cost; The method is simple and reliable, is applicable to large-scale production application.
Described silkworm is silkworm or Antherea pernyi Guerin-Meneville, Pupa bombycis is silkworm pupa or Antheraea pernyi Geurin Meneville, described transgenic baculovirus is bombyx mori nuclear polyhydrosis virus BmNPV and Antheraea pernyi nuclear polyhedrosis virus ApNPV, transgenic baculovirus expression system also claims silkworm or Antherea pernyi Guerin-Meneville bioreactor, described transgenic baculovirus expression system refers to be cultivated in specific recombinant baculovirus or gene insertion silkworm or Pupa bombycis body, can the recombinant baculovirus implanted or gene be transcribed and be translated in silkworm or Pupa bombycis body, the bioactive substance that Nature creating is useful to the mankind, as biologics, biovaccine, health food etc.
Described silkworm grass and Cordyceps militaris (L.) Link. have another name called Cordyceps militaris (L.) Link. or Cordyceps militaris (Cordycepsmilitaris), refer to that the domestication of Chinese caterpillar fungus strain spore is inoculated into silkworm body or Pupa bombycis body, parasitic life is carried out with the material in silkworm body or Pupa bombycis body, finally stretching out silkworm body or Pupa bombycis body surface after Sum decomposition and organism that a kind of fungus of being formed and silkworm body or Pupa bombycis body case coexist by growing, is a kind of integration of edible and medicinal herbs fungus of preciousness.
Described antibiotic refers to the aztreonam of beta-lactam, cefoperazone sodium, woods can mould rope class lincomycin hydrochloride, aminoglycoside streptomycin or fluoroquinolone antibiotics ciprofloxacin, antibiotic using method is puncture injection, and antibiotic service time is five length of times and pupa time of silkworm.
Described antibiotic use amount is as follows: with silkworm or silkworm pupa for the consumption of every bar silkworm or every pupa during host is: aztreonam is 40 ~ 80 μ g, cefoperazone sodium is 60 ~ 100 μ g, lincomycin hydrochloride is 15 ~ 40 μ g, streptomycin is 5 ~ 15 μ g, ciprofloxacin 4 ~ 8 μ g; With Antherea pernyi Guerin-Meneville or Antheraea pernyi Geurin Meneville for the consumption of every bar silkworm or every pupa during host is: aztreonam is 80 ~ 160 μ g, cefoperazone sodium is 120 ~ 200 μ g, lincomycin hydrochloride is 35 ~ 65 μ g, streptomycin is 10 ~ 20 μ g, ciprofloxacin 8 ~ 16 μ g, antibiotic before use, by above-mentioned dosage and transgenic baculovirus liquid or silkworm Cordyceps liquid or Cordyceps militaris (L.) Link. fungus liquid according to injecting object dosage mixture.
Embodiment 1
Composite antibiotic is used to express the impact of AsiaI foot-and-mouth disease antigen combination gene P1-2A3C to silkworm pupa bioreactor
(1) experiment material: AsiaI foot-and-mouth disease antigen combination gene recombinant baculovirus rBmNPV (P1-2A3C) and AsiaI type foot and mouth disease ELISA detection kit are provided because of Lanzhou, academy of agricultural sciences veterinary institute by middle.
(2) amplification of recombinant virus: the BmN attached cell of getting 100 μ l recombinant virus infection normal growths, after infection 4 DEG C of preservations after cell floats in about 5 days.
(3) inoculation of virus: employing QiuFeng×BaiYu Silkworm varieties in conventional manner example is supported to being placed on small straw bundles to spin cocoons, and 25 DEG C of protections are to the 3rd day injection inoculation of pupating.
(4) antibiotic combinations and preparation: get aztreonam for injection 0.5g, adds the solution that 5mL normal saline becomes 100mg/mL, and hydrochloric acid woods can adopt injection finished product by mould rope: content is 600mg/2mL.Get 8.13mL normal saline and add 1.2mL aztreonam (concentration 100mg/mL)+0.17mL lincomycin hydrochloride (concentration 300mg/mL), cumulative volume is 9.5mL.
(5) preparation of injection virus liquid: the recombinant virus liquid 0.5mL getting cell amplification good joins in the antibiotic solution of above-mentioned 9.5mL, becomes viral dilution injection, matching while using, every Pupa bombycis injects 5 μ L about containing 105pfu recombinant virus.Be equivalent to that every pupa injection aztreonam is 60 μ g, hydrochloric acid woods can mould rope be 25 μ g.
The check plot of adding composite antibiotic trial zone and only adding normal saline is established in test, and 50 Pupa bombycises are tested in every district, and in triplicate, before vaccinal injection, Pupa bombycis is sterilized with 0.3% effective chlorine bleaching powder and uses clear water rinsing.
Every Pupa bombycis vaccinal injection 5 μ L recombinant virus rBmNPV (Bm-P12A3C) diluent (being about 1.O × 105pfu containing recombinant virus), then express 120 hours at being placed in 25 DEG C, the incidence rate of investigation antibacterial deteriorated blood pupa, collect only morbidity viral disease and express the Pupa bombycis of product ,-20 DEG C frozen in order to detect expression efficiency.
(6) detection of antigen: every variety plot, each trial zone is in male and female 1: 1 ratio randomization Pupa bombycis, refiner grinding pupal cell, PBS dilution is added by 1: 10, after centrifuging and taking supernatant, the double antibody sandwich ELISA that reference reagent box description carries out AsiaI type foot-and-mouth disease antigen detects, and calculates the meansigma methods repeated for three times.
Experimental result is shown in Table 1:
The generation quantity of table 1 deteriorated blood pupa and ELISA method thereof measure the expression values (OD492nm) of P1-2A3C gene
Result shows, and the generation quantity of adding the trial zone antibacterial deteriorated blood pupa of composite antibiotic is antibiotic far less than not adding, and the expression no significant difference of the mNPV (P1-2A3C) of two kinds of process.
Embodiment 2
Compound antibiosis rope is adopted Antheraea pernyi Geurin Meneville to be produced to the impact of Cordyceps militaris (L.) Link.
(1) experiment material: artificial Chinese caterpillar fungus is produced bacterial strain ZUYCC and provided by Sheng Ke institute of Zhejiang University.
(2) strain cultivation: culture medium prescription is by peptone 20g, glucose 20g, NaH2PO41g, KH2PO41.5g, milk powder 5g, VB10.01g, and adding distil water is to 1000ml.Get this formula 100ml, load in 300ml triangular flask, add tampon disinfection inoculation, shaken cultivation 5 ~ 7d.
(3) Pupa bombycis is the live body Antheraea pernyi Geurin Meneville of commercially available rigidifying pupa
(4) antibiotic combinations and preparation: get injection cefoperazone sodium 1g, add the solution that 5mL normal saline becomes 200mg/mL, lincomycin hydrochloride adopts injection finished product: content is 600mg/2mL.Get 9.03mL normal saline to add 0.8mL cefoperazone sodium (concentration 200mg/mL) and add 0.17mL lincomycin hydrochloride (concentration 300mg/mL), cumulative volume is 10.0mL.
(5) injection inoculation bacterium liquid preparation: get strain cultivation liquid 90 and enter the antibiotic solution that ml adds the above-mentioned 10mL prepared, become containing antibiotic vaccinal injection liquid, matching while using, every Antheraea pernyi Geurin Meneville injection 0.1mL inoculates bacterium liquid, and being equivalent to every pupa injection cefoperazone sodium 160 μ g, lincomycin hydrochloride is 50 μ g.
The check plot of adding composite antibiotic trial zone and only adding normal saline is established in test, and 50 Pupa bombycises are tested in every district, and in triplicate, before vaccinal injection, Antheraea pernyi Geurin Meneville is sterilized with 0.3% effective chlorine bleaching powder and uses clear water rinsing.
Adopt injecting method by liquid-spawn inoculation in Antheraea pernyi Geurin Meneville body cavity, every Antheraea pernyi Geurin Meneville vaccinal injection 0.1mL Cordyceps militaris strain.
After injection inoculation, condition of culture according to a conventional method, and the cordycepin in Cordyceps militaris (L.) Link.sporophore and Cordyceps Determination of Adenosine adopt high performance liquid chromatography (with reference to Pharmacopoeia of People's Republic of China .2000 version annex VID).
Experimental result is shown in Table 2:
The experiment effect of table 2 cordyceps different vaccination method
Result shows, and the generation quantity of trial zone antibacterial deteriorated blood pupa of adding composite antibiotic is antibiotic far less than not adding, and success ratio of inoculation (hardening ratio) is high, and cordycepin in Cordyceps militaris (L.) Link.sporophore and Cordyceps adenosine content are without significant difference.
Embodiment 3
Composite antibacterial element is adopted silkworm to be produced to the impact of silkworm grass
(1) test material: artificial Chinese caterpillar fungus is produced bacterial strain ZUYCC and provided by Sheng Ke institute of Zhejiang University.
(2) strain cultivation: culture medium prescription is by peptone 20g, glucose 20g, NaH2PO41g, KH2PO41.5g, milk powder 5g, VB10.01g, and adding distil water is to 1000ml.Get this formula 100ml, load in 300ml triangular flask, add tampon disinfection inoculation, shaken cultivation 5 ~ 7d.
(3) antibiotic combinations and preparation: get injection cefoperazone sodium 1g, add the solution that 5mL normal saline becomes 200mg/mL, lincomycin hydrochloride adopts injection finished product: content is 600mg/2mL.Get 9.03mL normal saline to add 0.8mL cefoperazone sodium (concentration 200mg/mL) and add 0.17mL lincomycin hydrochloride (concentration 300mg/mL), cumulative volume is 10.0mL.
(4) injection inoculation bacterium liquid preparation: get strain cultivation liquid 90 and enter the antibiosis rope solution that ml adds the above-mentioned 10mL prepared, become containing antibiotic vaccinal injection liquid, matching while using, every Antheraea pernyi Geurin Meneville injection 0.1mL inoculates bacterium liquid.Being equivalent to every pupa injection cefoperazone sodium 160 μ g, lincomycin hydrochloride is 50 μ g.
The check plot of adding composite antibiotic trial zone and only adding normal saline is established in test, and 50 boss silkworms are tested in every district, in triplicate.Inoculation is started when silkworm larva is developed to the 2nd day 5 ages.Adopt injecting method by liquid-spawn inoculation in silkworm larva body cavity, every boss silkworm vaccinal injection 0.1mL silkworm grass liquid spawn.
After injection inoculation, condition of culture according to a conventional method, and the content of the cordycepin profit Cordyceps adenosine in silkworm Cordyceps sporophore adopts high effective liquid chromatography for measuring (with reference to Pharmacopoeia of People's Republic of China .2000 version annex VID).
Experimental result is shown in Table 3:
The experiment effect of table 3 silkworm silkworm grass different vaccination method
Result shows, and the generation quantity of trial zone antibacterial deteriorated blood pupa of adding composite antibiotic is antibiotic far less than not adding, and connects sharp success rate (hardening ratio) high, and cordycepin in silkworm Cordyceps sporophore and Cordyceps adenosine content are without significant difference.
The present invention adopts existing antibiotic interworking technology, effective antibiotics is selected to mix with appropriate ratio, the ratio that bacteriological infection effectively improves health hosts silkworm or Pupa bombycis can be reduced, significantly reduce silkworm or Pupa bombycis mortality rate, improving exogenous gene producing the production efficiency of silkworm grass or Cordyceps militaris (L.) Link. using silkworm or Pupa bombycis as carrier as the expression efficiency of expressing when cutting out body and raising using silkworm or Pupa bombycis, reducing production cost; The method is simple and reliable, is applicable to large-scale production application.
Above the better embodiment of this patent is explained in detail, but this patent is not limited to above-mentioned embodiment, in the ken that one skilled in the relevant art possesses, various change can also be made under the prerequisite not departing from this patent aim.
Claims (9)
1. one kind adopts the method reducing bacteriological infection when silkworm or Pupa bombycis production biological preparation, it is characterized in that, be adopt utilize transgenic baculovirus expression system to produce biological preparation using silkworm or Pupa bombycis as host cell or utilize silkworm or Pupa bombycis to produce silkworm grass or Cordyceps militaris (L.) Link. product as host cell time, by different types of for 2-3 kind antibiotic interworking conbined usage in host, bacterial disease is suppressed.
2. reduce the method for bacteriological infection when employing silkworm according to claim 1 or Pupa bombycis produce biological preparation, it is characterized in that, described silkworm is silkworm or Antherea pernyi Guerin-Meneville, and Pupa bombycis is silkworm pupa or Antheraea pernyi Geurin Meneville.
3. when employing silkworm according to claim 1 or Pupa bombycis produce biological preparation, reduce the method for bacteriological infection, it is characterized in that, described transgenic baculovirus is bombyx mori nuclear polyhydrosis virus BmNPV and Antheraea pernyi nuclear polyhedrosis virus ApNPV, described transgenic baculovirus expression system refers to be cultivated in specific recombinant baculovirus or gene insertion silkworm or Pupa bombycis body, can the recombinant baculovirus implanted or gene be transcribed and be translated, the bioactive substance that Nature creating is useful to the mankind in silkworm or Pupa bombycis body.
4. when employing silkworm according to claim 1 or Pupa bombycis produce biological preparation, reduce the method for bacteriological infection, it is characterized in that, described silkworm grass and Cordyceps militaris (L.) Link. refer to that the domestication of Chinese caterpillar fungus strain spore is inoculated into silkworm body or Pupa bombycis body, carrying out parasitic life with the material in silkworm body or Pupa bombycis body, finally to stretch out silkworm body or Pupa bombycis body surface after Sum decomposition and organism that a kind of fungus of being formed and silkworm body or Pupa bombycis body case coexist by growing.
5. when employing silkworm according to claim 1 or Pupa bombycis produce biological preparation, reduce the method for bacteriological infection, it is characterized in that, described antibiotic refers to the aztreonam of beta-lactam, cefoperazone sodium, lincomycin class lincomycin hydrochloride, aminoglycoside streptomycin or fluoroquinolone antibiotics ciprofloxacin.
6. reduce the method for bacteriological infection when employing silkworm according to claim 1 or Pupa bombycis produce biological preparation, it is characterized in that, described antibiotic using method is puncture injection.
7. reduce the method for bacteriological infection when employing silkworm according to claim 1 or Pupa bombycis produce biological preparation, it is characterized in that, described antibiotic service time is five length of times and pupa time of silkworm.
8. when employing silkworm according to claim 1 or Pupa bombycis produce biological preparation, reduce the method for bacteriological infection, it is characterized in that, described antibiotic use amount is as follows: with silkworm or silkworm pupa for the consumption of every bar silkworm or every pupa during host is: aztreonam is 40 ~ 80 μ g, cefoperazone sodium is 60 ~ 100 μ g, lincomycin hydrochloride is 15 ~ 40 μ g, streptomycin is 5 ~ 15 μ g, ciprofloxacin 4 ~ 8 μ g; With Antherea pernyi Guerin-Meneville or Antheraea pernyi Geurin Meneville for the consumption of every bar silkworm or every pupa during host is: aztreonam is 80 ~ 160 μ g, head embraces that piperazine ketone sodium is 120 ~ 200 μ g, lincomycin hydrochloride is 35 ~ 65 μ g, streptomycin is 10 ~ 20 μ g, ciprofloxacin 8 ~ 16 μ g.
9. when employing silkworm according to claim 8 or Pupa bombycis produce biological preparation, reduce the method for bacteriological infection, it is characterized in that, described antibiotic before use, by above-mentioned dosage and transgenic baculovirus liquid or silkworm Cordyceps liquid or Cordyceps militaris (L.) Link. fungus liquid according to injecting object dosage mixture.
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| CN108220250A (en) * | 2017-12-18 | 2018-06-29 | 蔡国祥 | A kind of method for expanding numerous bombyx mori nuclear polyhydrosis virus |
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Application publication date: 20160406 |