CN101623315A - Antibacterial immune double-effect compound Chinese herb for skin ulcerative syndrome of apostichopus japonicus - Google Patents
Antibacterial immune double-effect compound Chinese herb for skin ulcerative syndrome of apostichopus japonicus Download PDFInfo
- Publication number
- CN101623315A CN101623315A CN200910017176A CN200910017176A CN101623315A CN 101623315 A CN101623315 A CN 101623315A CN 200910017176 A CN200910017176 A CN 200910017176A CN 200910017176 A CN200910017176 A CN 200910017176A CN 101623315 A CN101623315 A CN 101623315A
- Authority
- CN
- China
- Prior art keywords
- herb
- ulcerative syndrome
- skin
- skin ulcerative
- syndrome
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 208000025865 Ulcer Diseases 0.000 title claims abstract description 33
- 150000001875 compounds Chemical class 0.000 title claims abstract description 32
- 208000011580 syndromic disease Diseases 0.000 title claims abstract description 29
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 12
- 241000965254 Apostichopus japonicus Species 0.000 title abstract description 23
- 241000746375 Andrographis Species 0.000 claims abstract description 15
- 230000036039 immunity Effects 0.000 claims abstract description 14
- 241000411851 herbal medicine Species 0.000 claims description 23
- 239000003814 drug Substances 0.000 claims description 17
- 241000628997 Flos Species 0.000 claims description 14
- 229940079593 drug Drugs 0.000 claims description 4
- 239000004615 ingredient Substances 0.000 claims description 4
- 201000010099 disease Diseases 0.000 abstract description 21
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 21
- 230000000694 effects Effects 0.000 abstract description 10
- 241000251511 Holothuroidea Species 0.000 abstract description 6
- 239000000843 powder Substances 0.000 abstract description 6
- 238000009395 breeding Methods 0.000 abstract description 5
- 230000001488 breeding effect Effects 0.000 abstract description 5
- 230000004083 survival effect Effects 0.000 abstract description 4
- 229940088710 antibiotic agent Drugs 0.000 abstract description 3
- 239000003242 anti bacterial agent Substances 0.000 abstract description 2
- 241000205585 Aquilegia canadensis Species 0.000 abstract 1
- 208000035240 Disease Resistance Diseases 0.000 abstract 1
- 241000334160 Isatis Species 0.000 abstract 1
- 241000212322 Levisticum officinale Species 0.000 abstract 1
- 239000001645 levisticum officinale Substances 0.000 abstract 1
- 239000002574 poison Substances 0.000 abstract 1
- 231100000614 poison Toxicity 0.000 abstract 1
- 210000003491 skin Anatomy 0.000 description 23
- 238000012360 testing method Methods 0.000 description 14
- 230000002265 prevention Effects 0.000 description 10
- 230000001717 pathogenic effect Effects 0.000 description 9
- GZCWLCBFPRFLKL-UHFFFAOYSA-N 1-prop-2-ynoxypropan-2-ol Chemical compound CC(O)COCC#C GZCWLCBFPRFLKL-UHFFFAOYSA-N 0.000 description 8
- 102000013563 Acid Phosphatase Human genes 0.000 description 8
- 108010051457 Acid Phosphatase Proteins 0.000 description 8
- 241000208340 Araliaceae Species 0.000 description 8
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 8
- 235000003140 Panax quinquefolius Nutrition 0.000 description 8
- 235000008434 ginseng Nutrition 0.000 description 8
- 244000052769 pathogen Species 0.000 description 8
- 208000024891 symptom Diseases 0.000 description 8
- 230000008901 benefit Effects 0.000 description 7
- 239000012530 fluid Substances 0.000 description 7
- 230000005764 inhibitory process Effects 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 206010030110 Oedema mouth Diseases 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- 230000001408 fungistatic effect Effects 0.000 description 5
- 230000036541 health Effects 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- 206010003694 Atrophy Diseases 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 4
- 241000519590 Pseudoalteromonas Species 0.000 description 4
- 241000589776 Pseudomonas putida Species 0.000 description 4
- 230000037444 atrophy Effects 0.000 description 4
- 230000003115 biocidal effect Effects 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 231100000397 ulcer Toxicity 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000003053 immunization Effects 0.000 description 3
- 238000002649 immunization Methods 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 210000003097 mucus Anatomy 0.000 description 3
- 229940126701 oral medication Drugs 0.000 description 3
- 229920003023 plastic Polymers 0.000 description 3
- 230000003449 preventive effect Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000005070 sampling Methods 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- 238000009631 Broth culture Methods 0.000 description 2
- 229920000742 Cotton Polymers 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 229920001503 Glucan Polymers 0.000 description 2
- 244000068988 Glycine max Species 0.000 description 2
- 235000010469 Glycine max Nutrition 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 241001135311 Pseudoalteromonas nigrifaciens Species 0.000 description 2
- 241000607598 Vibrio Species 0.000 description 2
- 241000607618 Vibrio harveyi Species 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 208000022531 anorexia Diseases 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 206010061428 decreased appetite Diseases 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 230000002766 immunoenhancing effect Effects 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000002844 melting Methods 0.000 description 2
- 230000008018 melting Effects 0.000 description 2
- 231100000957 no side effect Toxicity 0.000 description 2
- 210000000496 pancreas Anatomy 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 230000003285 pharmacodynamic effect Effects 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 208000017520 skin disease Diseases 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- PKDBCJSWQUOKDO-UHFFFAOYSA-M 2,3,5-triphenyltetrazolium chloride Chemical compound [Cl-].C1=CC=CC=C1C(N=[N+]1C=2C=CC=CC=2)=NN1C1=CC=CC=C1 PKDBCJSWQUOKDO-UHFFFAOYSA-M 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 208000031295 Animal disease Diseases 0.000 description 1
- 241001517488 Clavus Species 0.000 description 1
- 241000756943 Codonopsis Species 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 239000008589 Cortex Fraxini Substances 0.000 description 1
- 241000238557 Decapoda Species 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 description 1
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 1
- 241001619461 Poria <basidiomycete fungus> Species 0.000 description 1
- 241001148079 Vibrio splendidus Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 230000002421 anti-septic effect Effects 0.000 description 1
- 210000000436 anus Anatomy 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 239000010231 banlangen Substances 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 230000003559 chemosterilizing effect Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 239000003640 drug residue Substances 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000009313 farming Methods 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 239000004459 forage Substances 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000005541 medical transmission Effects 0.000 description 1
- LGZXYFMMLRYXLK-UHFFFAOYSA-N mercury(2+);sulfide Chemical compound [S-2].[Hg+2] LGZXYFMMLRYXLK-UHFFFAOYSA-N 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 238000011020 pilot scale process Methods 0.000 description 1
- 239000002985 plastic film Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000011085 pressure filtration Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000000384 rearing effect Effects 0.000 description 1
- 239000012744 reinforcing agent Substances 0.000 description 1
- 238000009738 saturating Methods 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 235000015170 shellfish Nutrition 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 235000011121 sodium hydroxide Nutrition 0.000 description 1
- UAJTZZNRJCKXJN-UHFFFAOYSA-M sodium;2-dodecoxy-2-oxoethanesulfonate Chemical compound [Na+].CCCCCCCCCCCCOC(=O)CS([O-])(=O)=O UAJTZZNRJCKXJN-UHFFFAOYSA-M 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000003206 sterilizing agent Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses an antibacterial immune double-effect compound Chinese herb for skin ulcerative syndrome of apostichopus japonicus. The herb comprises the following components: common andrographis herb, dyer woad leaf, honeysuckle and szechuan lovage rhizome; the weight ratio of the components is 2:1:3:2; and the herb is powered, wherein granularity is 200 mesh. The herb is orally taken to prevent and cure the skin ulcerative syndrome of apostichopus japonicus, wherein each kilogram feed is added with 20g of herb powder for the mild sea cucumber and each kilogram feed is added with 30g of herb powder for the critical sea cucumber. When in use, the herb is weighed according to required dosage and is fed after being evenly stirred with stichopus japonicus feed one time per day, wherein continues 20 days are a period of treatment. The Chinese herb is special for the skin ulcerative syndrome of apostichopus japonicus, which can be used for the young sea cucumbers and the adult sea cucumbers, has good protection effect during the peak period of the skin ulcerative syndrome, and has cure rate high to 80-90% and survival rate high to more than 85% for the apostichopus japonicus having the skin ulcerative syndrome within 2-3 weeks. The compound herb has a double effect for immunity and disease resistance, has no poison and side effect, convenient use, can reduce or replace the use of antibiotics, and is suitable for breeding the young sea cucumbers and controlling diseases during breeding and producing.
Description
Technical field:
The invention belongs to control and culture the special-purpose medicaments of Radix Morinae Bulleyanae major disease, is a kind of antibacterial immunity double-effect compound Chinese herbal medicine to the Radix Morinae Bulleyanae skin ulcerative syndrome.
Background technology:
At the beginning of 21 century, apostichopus japonicus culture is coastal flourish in northern China, and annual value of production surpasses 20,000,000,000 yuan, becomes the main body of the 5th sea-farming tide of China.Because the apostichopus japonicus culture scale is expanded rapidly, supporting technologies such as disease control relatively lag behind, add the operation lack of standardization in the production process, cause the disease problem to be on the rise, annual due to illness evil break out cause year economic loss above 3,000,000,000 yuan, wherein the harm that causes with Radix Morinae Bulleyanae " skin ulcerative syndrome " is the most serious, is the most important diseases that causes China's Radix Morinae Bulleyanae large tracts of land death.
Be treatment and prevention aquatic animal disease, the tradition method of preventing and treating is to use chemosterilant, antibiotic, and the history in existing more than 30 year; Because of life-time service, not only the Drug resistance of cause of disease is more and more obvious, and its drug residue also is on the rise, and causes disease to be difficult to cure, and aquatic product quality safety is affected; Vaccine also is effective way of prevention animal infectious disease, but defective such as targetedly strong, that sphere of action is little, wait invertebrates (as prawn, Radix Morinae Bulleyanae, shellfish etc.) effect limited for the low of main dependence nonspecific immunity system, and use operation also very inconvenient, be difficult to apply.
Before the present invention forms, mainly concentrate in the research of aspects such as epidemiology, nosetiology and histopathology about the report of culturing Radix Morinae Bulleyanae " skin ulcerative syndrome ".The inventor has taken the lead in carrying out the research of China's Stichopus japonicus disease, and has named breed Stichopus japonicus major disease " skin ulcerative syndrome " in 2004.Skin ulcerative syndrome is commonly called as " change skin disease ", and its cardinal symptom comprises: anorexia, shake the head, swollen mouth, row are dirty, mouthful turgescence canker, health atrophy, body surface large tracts of land ulcer, mortality rate reaches 80% in 1-2 week, its disease transmission is strong, involves wide.The pathogenic former multiformity that has of this disease, that has reported has 4 kinds, be respectively Vibrio splindidus (Vibrio splendidus), Pseudoalteromonas (Pseudoalteromona nigrifaciens), Vibro harveyi (Vibrio harveyi), pseudomonas putida (Pseudomonas putida), its disease symptom and main pathogen see table 1 for details.
Table 1: the disease symptom and the main pathogen of culturing Radix Morinae Bulleyanae " skin ulcerative syndrome ":
| Sampling time | The sampling position | Disease symptom | Pathogen | The intelligencer |
| 2003.04 | Area, Qingdao two morbidities plant | Infect the Radix Morinae Bulleyanae anorexia, shake the head, swollen mouth, row are dirty, mouthful turgescence canker, health atrophy, body surface large tracts of land ulcer, the mortality rates in indivedual ginsengs pond reach 80% in 1-2 week | Vibrio splindidus (V.plendidus) | Zhang Chunyun, Wang Yingeng, Rong Xiaojun |
| 2004.11-2005.01 | Mouping, Shandong Province, Jimo, area, Jiangnan 3 families morbidity apostichopus japonicus culture field | Initial stage infects Radix Morinae Bulleyanae and shakes the head oral area swelling, most of then dirty phenomenon of appearance row of attending a meeting of stinging; Infect Radix Morinae Bulleyanae health atrophy, stiff mid-term, small size ulcer appears earlier in food portion, forms the blue and white speckle; The body wall ulcer place of terminal infection Radix Morinae Bulleyanae increases, and the epidermis large tracts of land is rotted, and at last molten is nasal mucus shape colloid | Pseudoalteromonas (P.nigrifaciens) | Wang Yingeng, square wave, Zhang Chunyun, Rong Xiaojun |
| Spring in 2005 | The Jincheng Town, Laizhou City, Shandong Province | On the skin at oral area of the ill Radix Morinae Bulleyanae of their early stage or anus position small particles is arranged, the part Radix Morinae Bulleyanae has the phenomenon of telling intestinal, and the Radix Morinae Bulleyanae activeness reduces; Then the scope of this white point enlarges rapidly, and the phenomena of mortality will appear in Radix Morinae Bulleyanae in 3 days; Radix Morinae Bulleyanae all rots to become a pile white nasal mucus sample mucus in 5 days | Pseudoalteromonas (P.nigrifaciens) | Meng Qingguo, Wu Liuji, Wu Xinzhong, Yang Jianbo |
| 2006.01-2006.05 | Jimo City, Qingdao and Chengyang District | Swollen mouth, shake the head, arrange dirty, body surface blue and white and fester, the health atrophy, final disintegrate dead | Pseudomonas putida (P.putida) | Wang Yingeng, yellow Hua Wei |
| 2006.07-2006.10 | Jimo City, Qingdao and Chengyang District | Swollen mouth, dirty, the health micro of row, body surface occur blue and white fester disintegrate then dead.Dissect to find not arrange dirty disease and join its intestinal and be Chinese red, the body inwall is creamy white and is close to the shape that festers | Vibro harveyi (V. harveyi) | Wang Yingeng, yellow Hua Wei |
Up to now, domestic research report at Radix Morinae Bulleyanae " skin ulcerative syndrome " control medicine mainly is an antibiotic.Carry out the relevant report of oral medication with Chinese herbal medicine, rarely seen have Li Hua etc. " Chinese herbal compound preparation of treatment causative pathogen skin ulcerative syndrome of Apostichopus japonicus " (number of patent application: 200810012641.3), do not see that this prescription has effect and the relevant report that improves Stichopus japonicus immunity and pre-chemical defence skin disease.
Summary of the invention:
The object of the invention provides a kind of compound Chinese herb, solve the technical problem of its compound Chinese herb, make the use of this compound recipe can effectively improve Radix Morinae Bulleyanae immunity, and possess the function of the multiple pathogen of inhibition Radix Morinae Bulleyanae " skin ulcerative syndrome " simultaneously, improve the apostichopus japonicus culture survival rate.
The technical solution adopted in the present invention is:
Ingredient and proportioning are: ingredient is: Herba Andrographis, Folium Isatidis, Flos Lonicerae and Rhizoma Chuanxiong, medicine are Powdered, granularity 200 orders; Medicine is by weight ratio: 2: 1: 3: 2.
Take mode: oral.
Drug dose is: oral prevention or treat light disease consumption, and the per kilogram feedstuff adds 20g medicated powder, and oral medication serious symptom consumption is that the per kilogram feedstuff adds 30g medicated powder.
Using method is: when oral prevention and treatment use, all take by weighing medicine according to above-mentioned consumption, throw something and feed after stirring with holothurian feed, throw something and feed every day 1 time, continuous 20 days is a course of treatment.
Compound recipe of the present invention has following characteristics:
1, the present invention is developing on the multiple former basis of carrying out folk prescription, compound recipe pharmacodynamic analysis and Stichopus japonicus immunization of causing a disease skin ulcerative syndrome, is having the immunity of the Radix Morinae Bulleyanae of raising, high-efficiency antimicrobial economic benefits and social benefits characteristics.
2, the Radix Morinae Bulleyanae that the present invention prepared " skin ulcerative syndrome " special-purpose medicaments is joined the phase and is become the ginseng phase all can use the children, can reach more than 80% the cure rate of ill Radix Morinae Bulleyanae, and survival rate reaches more than 85%.
3, use operational approach easy, only need medicine to be added to throw something and feed in the feedstuff in proportion to get final product, be applicable to that children's ginseng is cultivated and the control of breeding production disease.
4, the compound recipe composition adopts Chinese herbal medicine green, that have no side effect, and is cheap.Use the present invention to reduce or alternative antibiotic use, the commercial quality and the food safety of culturing Radix Morinae Bulleyanae had guaranteeing role.
Description of drawings:
Fig. 1: the variation of coelomic fluid neutral and alkali phosphatase (AKP);
Fig. 2: the variation of acid phosphatase in the coelomic fluid (ACP).
The specific embodiment:
The present invention is developing on the multiple former basis of carrying out folk prescription, compound recipe pharmacodynamic analysis and Stichopus japonicus immunization of causing a disease skin ulcerative syndrome, is having antibacterial immunity economic benefits and social benefits characteristics.
Be described in detail technology contents of the present invention in conjunction with the accompanying drawings with embodiment below:
One, determining of the screening of ingredient of the present invention and proportioning:
1, the screening of medicine and processing:
According to herbal nature, select Rhizoma Atractylodis, Radix Et Rhizoma Rhei, Herba Portulacae, Radix Glycyrrhizae, Herba Andrographis, Folium Isatidis, Herba Houttuyniae, Radix Angelicae Sinensis, Flos Lonicerae, Cortex Acanthopancis, the Cortex Eucommiae, Herba Epimedii, Cortex Fraxini, Rhizoma Chuanxiong, Radix Bupleuri, Poria, Cortex Phellodendri, Radix Sanguisorbae, Radix Codonopsis, Galla Chinensis, Radix Isatidis and 24 kinds of Chinese herbal medicine dry powder of Radix Scutellariae for use, number consecutively is H-1, H-2, H-3......H-24.
Chinese herbal medicine dry powder respectively takes by weighing 5g after sieving by 200 orders, adds about 50 ℃ of distilled water respectively, and promptly 350mL soaked after 2 hours, was heated to boiling with big fire, and littler fire boils 90min; When being concentrated into 50mL, leach medicinal liquid with 8 layers of antiseptic gauze, use double-deck filter paper pressure filtration at last, promptly make the Chinese herbal medicine stock solution of 0.1g/mL concentration, 121 ℃ of sterilization 20min are standby.
With the 96 aseptic hole ELISA Plate (transparent plastic sheets that 96 holes are arranged, the big swept volume of every Kongzui is about 250 μ L) wherein 7 wells add and measure Chinese herbal medicine stock solution (0.1g/mL), (TSB is the pancreas peptone soybean broth culture medium with liquid TSB, available from Beijing overpass technology company limited) carry out 2 times of dilutions of going forward one by one, add 5 μ L, 0.5% red tetrazolium indicator in every then hole, be used to whether indicate bacterial growth.Shaking table is cultivated 6-8h in 27 ℃ of wet boxes, and liquid presents redness in its corresponding ELISA Plate hole of person that has the bacteria growing, and no change color person's minimum inhibitory concentration is the MIC value.
For influencing judgement person because of Chinese herbal medicine band color itself, dip in the mixed liquor of getting in the ELISA Plate hole with aseptic cotton carrier and coat that (TSB is the pancreas peptone soybean broth culture medium on the TSB flat board, both can be configured to the solid plate culture medium, also can be configured to fluid medium), directly take culture method to observe drug effect and minimum inhibitory concentration that whether bacterial growth judges medicine, see Table 2.The result shows that folk prescription Chinese herbal medicine H-5 (Herba Andrographis), H-6 (Folium Isatidis), H-9 (Flos Lonicerae) and H-14 (Rhizoma Chuanxiong) have good fungistatic effect to 4 strain Stichopus japonicus skin ulcerative syndrome pathogen, and its MIC value is respectively 6.25,7.81,5.86 and 3.52mg/L.
Show the minimum inhibitory concentration (mg/mL) of 2:24 kind folk prescription Chinese herbal medicine water decoction to 4 kinds of antibacterials:
2, inhibition zone is measured:
Cut-off is the flat pair of dish of about 90mm directly, injection is through the TSB of heating and melting 7-9mL, be evenly distributed at the bottom of the dish, place superclean bench, treat that it solidifies after, insert internal diameter 6.0 ± 0.1mm at each flat pair of medium branch of dish, external diameter 7.8 ± 0.1mm Oxford cup, pour 15-17mL again into, solidify back reuse sterilization tweezers the Oxford cup is taken out, just form culture medium flat plate with holes through the TSB of heating and melting.After treating that flat board is cooled to room temperature, with bacteria suspension (1 * 10
7Cfu/mL) with the evenly coating on perforated flat plate of sterilization cotton swab, in the hole, add 100 μ L Chinese herbal medicine stock solutions then, every kind of Chinese herbal medicine is done 2 parallel group simultaneously, measure record inhibition zone size (the inhibition zone size reflects the size of medicine fungistatic effect), get its meansigma methods and (average and to eliminate experimental error.Be the experiment basis of selecting 4 kinds of folk prescription Chinese herbal medicine).By the test of vitro inhibition " skin ulcerative syndrome " pathogen, from 24 kinds of Chinese herbal medicine, filter out 4 kinds of folk prescription Chinese herbal medicine of best fungistatic effect, promptly Herba Andrographis, Folium Isatidis, Flos Lonicerae, Rhizoma Chuanxiong see Table 3.
The inhibition zone of table 3:24 kind folk prescription Chinese herbal medicine water decoction is measured:
Unit (mm)
Annotate: because the hole of the cup-shaped one-tenth in Oxford is approximately 10mm, so antibacterial circle diameter≤10mm duration is 0.
And then design orthogonal test, see Table 4, compound Chinese herb H-5 (Herba Andrographis), H-6 (Folium Isatidis), H-9 (Flos Lonicerae), when H-14 (Rhizoma Chuanxiong) ratio is respectively 20g, 10g, 30g and 20g, its average diameter of inhibition zone reaches maximum respectively, is 15.25mm, 12.67mm, 13.50mm and 13.25mm.The best proportioning of compound recipe is H-5: H-6: H-9: H-14=2: 1: 3: 2.
Table 4: compound compatibility and fungistatic effect analytical table:
Annotate: MIC represents the MIC meansigma methods, and BC represents the antibacterial circle diameter meansigma methods.
Carry out extracorporeal bacteria inhibitor test again, the result proves that (H-5: H-6: H-9: H-14=2: 1: 3: 2) is obvious to the fungistatic effect of 4 kinds of pathogenic bacterium for this compound recipe, sees Table 5.
Table 5: compound Chinese herb is to the extracorporeal bacteria inhibitor test of 4 kinds of antibacterials:
"+" antibacterial "-" of representing not grow under this concentration is represented the antibacterial that grown under this concentration.
Two, the enforcement experiment and the effect of medicine:
1, immunoenhancement result detects test group:
Select healthy Stichopus japonicus individuality in Tian Yuan Aquatic product company limited holothruian cultures field, Yantai, Shandong in October, 2007, and the 30-90g/ head is raised in the white plastic case of about 35L size, adds water 20L.Every rearing in box is supported 6, raises 3 casees for every group, and test is divided into 6 groups, and the test group normal feedstuff is joined precious board children ginseng special compound feed for the benefit that Dalian Feng Yuan reaches the production of bait company limited.Go into Herba Andrographis, Folium Isatidis, Flos Lonicerae in every kg feedstuff, (mass ratio is Rhizoma Chuanxiong: 2: 1: 3: 2) compound Chinese herb, set 0.5%, 1%, 3%3 drug level gradient, each experimental group is its positive controls with 0.5%, 1% and 3% β-1,3-glucosan respectively, what only throw something and feed normal feedstuff is the blank group, and the forage volume of throwing something and feeding is thrown something and fed by 1% of Radix Morinae Bulleyanae body weight.The water source is the deep-well sea water, and average water temperature is 16 ℃.Morning, siphonage was inhaled the end, changed water once, threw something and fed once at dusk.
Support 1 week, not feeding in preceding 2 days, the back 5 days normal feedstuff of throwing something and feeding before the test temporarily.After on-test, per 5 days once sampling, fish for 4 Radix Morinae Bulleyanaes, each extracts the 0.3mL coelomic fluid in centrifuge tube at every turn, measures these 2 indexs just relevant with Radix Morinae Bulleyanae immunity of centrifuge tube mesenteron liquid caustic soda acid phosphatase (AKP) and acid phosphatase (ACP) that finish in 3 hours.As shown in Figure 1, each is organized reinforcing agent there were significant differences to the Radix Morinae Bulleyanae immunoenhancement result.The high dose interpolation group (3%) of compound recipe was since the 5th day, and AKP raises by a small margin, the 10th day, the utmost point is significantly higher than blank group (P＜0.01), and the 20th day peaking is blank 5.5 times of organizing, be 4 times of initial value (the 0th day), begin after 25 days to descend, but still be higher than blank group.0.5% performance of middle dosage group 1% and low dose group steadily.As shown in Figure 2,3% pair of Radix Morinae Bulleyanae coelomic fluid of high dose interpolation group ACP influence is the most obvious, in the time of the 10th day, the high dose interpolation is organized just, and the utmost point is significantly higher than blank group (P＜0.01), be reduced to subsequently and be lower than blank group, but peaking the 25th day the time is significantly higher than blank group (P＜0.05), be significantly higher than other test group ACP same period value simultaneously, the utmost point is significantly higher than initial value (P＜0.01).In dosage group and low dose group to compare difference in duration of test ACP vigor and blank group not remarkable.Show that by immunization experiment compound Chinese herb is adding the immunity that can significantly improve Radix Morinae Bulleyanae under 3% dosage, promptly add the 30g compound Chinese herb in the per kilogram feedstuff and can obtain best immune effect.
2, safety of medicine property testing:
Divide 2 experimental grouies to carry out.Add 20% compound Chinese herb (20g Chinese herbal medicine/80g feedstuff) in experimental group 1 feedstuff, add 25% Chinese herbal medicine (25g Chinese herbal medicine/75g feedstuff) in experimental group 2 feedstuffs, the matched group basic feedstuff of only throwing something and feeding.Each experimental group is established 3 repeating groups, and each repeating groups has 6 Radix Morinae Bulleyanaes (30-90g/ head).Support temporarily before the experiment and do not throw feedstuff in 2 days.During experiment the Chinese herbal medicine of aforementioned proportion is added in the fluid feedstuff (the fluid feedstuff Stichopus japonicus of throwing something and feeding can directly ingest), stir and soak the Radix Morinae Bulleyanae of throwing something and feeding after 2 hours, apostichopus japonicus culture is in the white plastic case.Determine the feed situation of Radix Morinae Bulleyanae by the 2nd day Radix Morinae Bulleyanae bait surplus and feces situation.Duration of test runs 15 days.The result shows: each experimental group Radix Morinae Bulleyanae does not have any unusual condition, and each ratio is added medicine and all Radix Morinae Bulleyanae do not caused toxic effect, and Radix Morinae Bulleyanae ingests normally, well-grown.
Conclusion: when oral medication carries out diseases prevention and treatment, add compound Chinese herb 30g/kg feedstuff, can improve Stichopus japonicus immunity, and Stichopus japonicus is had no side effect.
Three, using this compound Chinese herb treats 6 different breeding fields, area, Shandong trouble " skin ulcerative syndrome " Radix Morinae Bulleyanae.Effect such as following table 6:
The clinical pilot-scale experiment of table 6 compound Chinese herb control skin ulcerative syndrome
| Time | The place | Symptom | Compound recipe proportioning and using method | The course of treatment | Effect |
| ??2007.11 | Yantai | The 50-120g Radix Morinae Bulleyanae, a little less than the ingestion ability, part body surface large tracts of land is festered | By Herba Andrographis: Folium Isatidis: Flos Lonicerae: Rhizoma Chuanxiong=2: 1: 3: 2 proportioning 3% companion raises the treatment of throwing something and feeding | 18 days | Cure rate 80% |
| ??2007.11 | Peng Lai | 2 ages, ginseng was shaken the head, | Herba Andrographis: Folium Isatidis: Flos Lonicerae: Rhizoma Chuanxiong | 20 days | Cure rate |
| Swollen mouth, body surface festers | =2: 1: 3: 2 proportioning 3% companion raises the treatment of throwing something and feeding | ??85％ | |||
| ??2007.12 | Qingdao | Young ginseng is shaken the head, and swollen mouth has self-dissolving | Herba Andrographis: Folium Isatidis: Flos Lonicerae: Rhizoma Chuanxiong=2: 1: 3: 2 proportioning 3% companion raises the treatment of throwing something and feeding | 14 days | Cure rate 80% |
| ??2008.1 | Weihai | It is mashed saturating that 50-120g ginseng, part are joined body surface, and part is arranged dirty | Herba Andrographis: Folium Isatidis: Flos Lonicerae: Rhizoma Chuanxiong=2: 1: 3: 2 proportioning 2% companion raises the treatment of throwing something and feeding | 20 days | Cure rate 90% |
| ??2008.3 | Sunshine | Young ginseng clavus is bald blunt, and part is arranged dirty | Herba Andrographis: Folium Isatidis: Flos Lonicerae: Rhizoma Chuanxiong=2: 1: 3: 2 proportioning 2% companion raises the treatment of throwing something and feeding | 17 days | Cure rate 90% |
| ??2008.2 | The Jiangnan | The skin ulcerative syndrome high-incidence season | Herba Andrographis: Folium Isatidis: Flos Lonicerae: Rhizoma Chuanxiong=2: 1: 3: 2 proportioning 2% companion raises the prevention of throwing something and feeding | 20 days | Do not see morbidity, preventive effect is remarkable |
| ??2008.2 | Peng Lai | The skin ulcerative syndrome high-incidence season | Herba Andrographis: Folium Isatidis: Flos Lonicerae: Rhizoma Chuanxiong=2: 1: 3: 2 proportioning 2% companion raises the prevention of throwing something and feeding | 20 days | Do not see morbidity, preventive effect is remarkable |
By a large amount of clinical prevention evidences: in the skin ulcerative syndrome high-incidence season, raise the prevention of throwing something and feeding with 2% companion, do not see morbidity, its preventive effect is good; Patients whose symptoms were less severe raise and throw something and feed with 2% companion, and the heavier person of symptom raises with 3% companion and throws something and feeds, and its cure rate is more than the 80-90% in the treatment 20 days of throwing something and feeding.Carried out the treatment pilot plant test in many plants in the clinical practice process, prevention effect is remarkable, has retrieved a large amount of economic losses for culturing unit.The present invention has started the precedent of utilizing Chinese herbal medicine to improve Stichopus japonicus immunity and effectively suppressing the skin ulcerative syndrome pathogen simultaneously, have immunity, disease-resistant economic benefits and social benefits effect, can reduce and replace antibiotic use, to solving major disease " skin ulcerative syndrome ", improve and culture survival rate and product quality, ensure that the breeding production stable and high yields has crucial meaning.
Claims (1)
1, a kind of antibacterial immunity double-effect compound Chinese herbal medicine of Radix Morinae Bulleyanae skin ulcerative syndrome is characterized in that its ingredient is: Herba Andrographis, Folium Isatidis, Flos Lonicerae and Rhizoma Chuanxiong; Medicine is by weight ratio: 2: 1: 3: 2; It is Powdered that medicine is, granularity 200 orders.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100171767A CN101623315B (en) | 2009-07-28 | 2009-07-28 | Antibacterial immune double-effect compound Chinese herb for skin ulcerative syndrome of apostichopus japonicus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100171767A CN101623315B (en) | 2009-07-28 | 2009-07-28 | Antibacterial immune double-effect compound Chinese herb for skin ulcerative syndrome of apostichopus japonicus |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101623315A true CN101623315A (en) | 2010-01-13 |
| CN101623315B CN101623315B (en) | 2011-01-26 |
Family
ID=41519410
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009100171767A Active CN101623315B (en) | 2009-07-28 | 2009-07-28 | Antibacterial immune double-effect compound Chinese herb for skin ulcerative syndrome of apostichopus japonicus |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101623315B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102206595A (en) * | 2011-03-31 | 2011-10-05 | 国家海洋局第一海洋研究所 | Apostichopus japonicus antagonistic bacterium and its application |
| CN103463389A (en) * | 2013-08-30 | 2013-12-25 | 兴化市恒威生物技术有限公司 | Medicinal composition for treating skin ulceration syndrome of sea cucumber and preparation method thereof |
| CN104815249A (en) * | 2015-05-20 | 2015-08-05 | 山东亚康药业股份有限公司 | Traditional Chinese medicine preparation for treating stichopus japonicus ulcerative syndrome with high cure rate and preparation method thereof |
| CN108949615A (en) * | 2018-06-25 | 2018-12-07 | 青岛农业大学 | Probiotics for the apostichopus japonicus Selenka feed that ferments |
-
2009
- 2009-07-28 CN CN2009100171767A patent/CN101623315B/en active Active
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102206595A (en) * | 2011-03-31 | 2011-10-05 | 国家海洋局第一海洋研究所 | Apostichopus japonicus antagonistic bacterium and its application |
| CN102206595B (en) * | 2011-03-31 | 2012-12-05 | 国家海洋局第一海洋研究所 | Apostichopus japonicus antagonistic bacterium and its application |
| CN103463389A (en) * | 2013-08-30 | 2013-12-25 | 兴化市恒威生物技术有限公司 | Medicinal composition for treating skin ulceration syndrome of sea cucumber and preparation method thereof |
| CN104815249A (en) * | 2015-05-20 | 2015-08-05 | 山东亚康药业股份有限公司 | Traditional Chinese medicine preparation for treating stichopus japonicus ulcerative syndrome with high cure rate and preparation method thereof |
| CN104815249B (en) * | 2015-05-20 | 2018-04-27 | 山东亚康药业股份有限公司 | A kind of Chinese medicine preparation and preparation method for treating stichopus japonicus skin ulcer syndrome |
| CN108949615A (en) * | 2018-06-25 | 2018-12-07 | 青岛农业大学 | Probiotics for the apostichopus japonicus Selenka feed that ferments |
| CN108949615B (en) * | 2018-06-25 | 2020-05-05 | 青岛农业大学 | Microecological preparation for fermenting stichopus japonicus feed |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101623315B (en) | 2011-01-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103141709B (en) | Carp feed and preparation method thereof | |
| CN106259097A (en) | A kind of ecological cultivation method of Cyprinus carpio | |
| CN101744924B (en) | Multiple-effect Chinese herbal medicine compound of bacterial diseases of turbot | |
| CN103349747A (en) | Traditional Chinese medicine preparation for preventing and curing paralichthys olivaceus and sea cucumber scuticoci liatlda diseases | |
| CN101623315B (en) | Antibacterial immune double-effect compound Chinese herb for skin ulcerative syndrome of apostichopus japonicus | |
| CN102524584B (en) | Composite pig feed additive for effectively preventing edema of pigs | |
| CN102771874A (en) | Natural food preservative and preparation method and applications thereof | |
| CN103830404A (en) | Traditional Chinese medicine composition for treating Chicken colibacillosis, preparation method of traditional Chinese medicine mixture and traditional Chinese medicine mixture | |
| DE202023100017U1 (en) | Novel drug for the prevention and treatment of bacterial mixed infections in freshwater fish | |
| CN102240323A (en) | Traditional Chinese medicine for controlling aeromonas hydrophila disease of aquatic animals and utilizing method thereof | |
| CN100589821C (en) | Method for preparing Plague-clearing toxin-removing oral liquid for treating infectious bursa of Fabricius disease of livestock and fowl | |
| CN101422419A (en) | Toilet water and manufacture method thereof | |
| CN106619887B (en) | Compound traditional Chinese medicine preparation for inhibiting and killing schbert unicellular bacteria, preparation method and application thereof | |
| CN101961399A (en) | Preparation method of insecticide for aquatic animals | |
| CN106689222A (en) | Traditional Chinese medicine disinfectant for preventing aquaculture bacteriosis and preparation method of disinfectant | |
| CN102935166A (en) | Chinese herbal medicine compound fish medicine | |
| CN104222527B (en) | The application process of belulinic acid Betulinic acid | |
| CN100571721C (en) | Long stalk Fructus Schisandrae Sphenantherae extract, Preparation Method And The Use | |
| CN102416050B (en) | Veterinary medicinal preparation for treating colibacillosis and preparation method thereof | |
| CN105055401A (en) | Application of chrysin in preparing cryptosporidium parvum resisting medicine | |
| CN102895546B (en) | Chinese herbal medicine compound formula for preventing and treating apostichopus japonicus balantidiasis | |
| CN105250387A (en) | Antibacterial Chinese herbal medicine compound for fishes as well as preparation method and application of antibacterial Chinese herbal medicine compound | |
| CN100509038C (en) | Compound decoction with astragalus root for birds and method for preparing same | |
| CN103919889B (en) | A kind of Chinese medicine composition strengthening turbot immunity | |
| CN102657729A (en) | Traditional Chinese medicine injection used in treatment of dairy cow mastitis and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Assignee: QINGDAO ZHONGREN PHARMACEUTICAL Co.,Ltd. Assignor: YELLOW SEA FISHERIES Research Institute CHINESE ACADEMY OF FISHERY SCIENCES Contract record no.: 2011990000391 Denomination of invention: Antibacterial immune double-effect compound Chinese herb for skin ulcerative syndrome of apostichopus japonicus Granted publication date: 20110126 License type: Exclusive License Open date: 20100113 Record date: 20110524 |
|
| EM01 | Change of recordation of patent licensing contract |
Change date: 20140821 Contract record no.: 2011990000391 Assignee after: QINGDAO ZHONGREN ANIMAL PHARMACEUTICAL CO.,LTD. Assignee before: QINGDAO ZHONGREN PHARMACEUTICAL Co.,Ltd. |
|
| LICC | Enforcement, change and cancellation of record of contracts on the licence for exploitation of a patent or utility model | ||
| EC01 | Cancellation of recordation of patent licensing contract | ||
| EC01 | Cancellation of recordation of patent licensing contract |
Assignee: QINGDAO ZHONGREN ANIMAL PHARMACEUTICAL CO.,LTD. Assignor: YELLOW SEA FISHERIES Research Institute CHINESE ACADEMY OF FISHERY SCIENCES Contract record no.: 2011990000391 Date of cancellation: 20221202 |
|
| EE01 | Entry into force of recordation of patent licensing contract | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20100113 Assignee: Fuzhou Tiankai Biotechnology Co.,Ltd. Assignor: YELLOW SEA FISHERIES Research Institute CHINESE ACADEMY OF FISHERY SCIENCES Contract record no.: X2022370010023 Denomination of invention: Antibacterial and Immune Compound Chinese Herbs for Stichopus japonicus Rot Skin Syndrome Granted publication date: 20110126 License type: Common License Record date: 20221206 |