CN103966122A - Providencia vermicola and application of providencia vermicola in preparing antibacterial drugs - Google Patents
Providencia vermicola and application of providencia vermicola in preparing antibacterial drugs Download PDFInfo
- Publication number
- CN103966122A CN103966122A CN201410159344.7A CN201410159344A CN103966122A CN 103966122 A CN103966122 A CN 103966122A CN 201410159344 A CN201410159344 A CN 201410159344A CN 103966122 A CN103966122 A CN 103966122A
- Authority
- CN
- China
- Prior art keywords
- vibrio
- preparation
- vibrio alginolyticus
- providencia vermicola
- vermicola
- 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
Landscapes
- Farming Of Fish And Shellfish (AREA)
- Fodder In General (AREA)
Abstract
The invention discloses providencia vermicola and application of providencia vermicola in preparing antibacterial drugs. The preservation number of providencia vermicola No. 22 is CCTCC No. M2013696. The providencia vermicola No. 22 has antagonism, so that the providencia vermicola No. 22 can be used for preparing preparation for preventing and controlling vibrio alginolyticus or preparing assistant hemolysis vibrio preparation; the preparation for preventing and controlling vibrio alginolyticus or the assistant hemolysis vibrio preparation can be used for beneficial microbial preparation to replace chemical medicine, improves water quality, and eliminates vibrio alginolyticus or assistant hemolysis vibrio in an aquatic product cultivation system, so that damage to aquatic product cultivation objects from the vibrio alginolyticus or assistant hemolysis vibrio is prevented, the development of the aquatic product cultivation industry is promoted, aquatic products are prevented from carrying the vibrio alginolyticus or assistant hemolysis vibrio, and health and safety of human beings are ensured.
Description
Technical field:
The invention belongs to aquaculture field, be specifically related to a strain activity resistent ingredient stability high, can be as Providence and the application in preparation antibacterials thereof of the Antagonistic Fungi of multiple bacterium.
Background technology:
Vibrio alginolyticus (Vibrio alginolyticus, V.alginolyticus) is subordinate to vibrionaceae (Vibrionaceae), Vibrio (Vibrio), and this bacterium is present in seawater, river mouth, Organisms.Vibrio alginolyticus has stronger pathogenic to cultivated animals such as fish, shrimp, shellfishes, can cause cultivated animals acute hemorrhagic septicemia.This disease, also claim Outbreak disease, vibriosis, ulcer in body surface etc., can infect multiple sea farming kind, as large yellow croaker, perch, lefteye flounder, malaber reefcod, black porgy, flat porgy, cabio, Crustin, japonicus, tigar prawn, Environment of Litopenaeus vannamei Low, clam, cuttlefish etc., it is the popular wider a kind of hydrocoles conditioned pathogen of China.This bacterium is Gram-negative tyrothricin, without gemma, pod membrane, Individual existence or tail end are connected to " C " or serpentine, for having a liking for salt, have a liking for warm nature, the raw vibrios in amphimicrobian sea, optimal temperature is 17～35 ℃, and growth optimum pH scope is 6.0～9.0, economize in recent years the explosive transmissible disease of cultured fishes shrimps in China Hainan, Guangdong, Fujian etc., multithread row and summer, a lot of reports are septicemia due to this bacterium, and sea farming disease control is more difficult than fresh water.In addition vibrio alginolyticus also has a large amount of reports to people pathogenic, can cause that people poisons by food, otitis etc.Vibrio alginolyticus is one of the main pathogenic fungi of sea farming kind fulminant transmissible disease.
The method of preventing and treating of this bacterium mainly contains: microbiotic control, herbal control, vaccine prevention and control, probiotic bacterium biological and ecological methods to prevent plant disease, pests, and erosion.
(1) microbiotic control.Microbiotic is that microbiotic has significant inhibition or killing action to pathogenic bacteria for treating the choice drug of bacterial infectious disease, studies confirm that the medicines such as sulfanilamide (SN)+TMP, Ciprofloxacin, furadantin have stronger fungistatic effect to vibrio alginolyticus.But be widely used along with antibiotic, different areas are because types of medicines is different with usage in recent years, produced a large amount of Resistant strain, fishery products drug residue etc. and threatened aquaculture and human health, fishery products resistance can escalated dose just be absorbed in vicious cycle again simultaneously.(2) herbal control.Herbal medicine is without resistance, noresidue, and non-environmental-pollution, can not cause drug-induced disease and fungistatic effect is obvious, more and more causes people's concern.Research generally shows that folk prescription is used the herbal medicine such as pomegranate rind, garden burnet, shizandra berry, rheum officinale, dark plum, Flos Caryophylli, the capsule of weeping forsythia, Root of Indigowoad to have very strong bacteriostasis, in actual production, the herbal medicine with bacteriostatic action is combined into compound, suitably utilize different herbal medicine mutually to act synergistically to prevent and treat the disease that vibrio alginolyticus causes, can better bring into play the bacteriostatic action of herbal medicine.But unreasonable, the abuse folk prescription of filling a prescription, the problem such as complete processing is backward, theoretical investigation is less, make blindly medication of a lot of culturists.(3) vaccine prevention and control.The vaccine using in aquaculture is mainly divided into two classes: a class is attenuated vaccine, and another kind of is inactivated vaccine.The cause of disease alive that attenuated vaccine contains attenuation, energy inducing specific antibody, and similar to natural infection; The vaccine of deactivation is comprised of as the toxin of bacteria lipopolysaccharide or deactivation some compositions of dead cause of disease or cause of disease, can stimulating animal immunity system.Its immunization ways has immersion, oral, injection and spraying etc., injection system best results but waste time and energy and damage shrimp body, and soaking vaccine requirement is large; The development of vaccine is in the starting stage, and attenuated vaccine also lacks security guarantee, preparation trouble, high in cost of production; Also lack simple and easy, efficient, economic route of inoculation.(4) probiotic bacterium prevention and control.Along with making antibiotic use, the appearance of resistance problem and the deterioration of environment be subject to increasing test, and probiotic bacterium relies on the advantage of non-toxic efficient, by improving water quality, in host internal secretion nutritive substance competition or there is attachment site, strengthen host immune power, improve disease resistance, reduce the sickness rate of disease, and then improve aquaculture output and economic benefit, just receiving increasing concern.At present, on aquatic products disease is controlled, use more probiotic bacterium for the composite reactive microbial preparation that milk-acid bacteria, Vibrio, genus bacillus, photosynthetic bacterium, yeast, Rhodopseudomonas etc. are single or multiple beneficial bacterium forms, be applied in the cultivation of fish, shrimp, crab, shellfish, molluscan cultivation and bait.
Summary of the invention:
The object of this invention is to provide No. 22, a kind of Providence (providencia vermicola), this bacterial strain can effectively be eliminated the vibrio alginolyticus in aquaculture system with the effect of Antagonistic Fungi, has solved fishery products in prior art and has carried the immunization issues of vibrio alginolyticus.
Providence of the present invention (providencia vermicola) is preserved in Chinese Typical Representative culture collection center (CCTCC), address No. 22 on December 24th, 2013: Wuhan, China Wuhan University, deposit number is CCTCC No.M2013696.
Providence of the present invention (providencia vermicola) has antagonistic action to multiple bacterium No. 22, therefore can be by No. 22, Providence of the present invention (providencia vermicola) for the preparation of in antibacterials, as prevention and control subtilis, chromobacterium violaceum preparation, particularly in prevention and control aquatic products vibrios preparation, as prevention and control aquatic products vibrio alginolyticus or Vibrio parahaemolyticus preparation.Be applicable to the aquatic animals such as fish, shrimp, can be used for preventing and treating the disease that vibrio alginolyticus causes.
Providence of the present invention (providencia vermicola) has antagonistic action No. 22, therefore can be by No. 22, Providence of the present invention (providencia vermicola) for the preparation of in prevention and control vibrio alginolyticus or Vibrio parahaemolyticus preparation, this prevention and control vibrio alginolyticus or Vibrio parahaemolyticus preparation can be used as the effective microorganism preparation of instead of chemical medicine, be used for improving water quality, remove vibrio alginolyticus or Vibrio parahaemolyticus in aquaculture system, thereby prevention vibrio alginolyticus or the harm of Vibrio parahaemolyticus to aquaculture object, promote the development of culture fishery, avoid fishery products to carry vibrio alginolyticus or Vibrio parahaemolyticus, ensure human health safety.
Providence of the present invention (providencia vermicola) is preserved in Chinese Typical Representative culture collection center (CCTCC), address No. 22 on December 24th, 2013: Wuhan, China Wuhan University, deposit number is CCTCC No.M2013696.
Accompanying drawing explanation:
Fig. 1 is the screening of Antagonistic Fungi;
Fig. 2 is halogen worm Protection figure, and wherein experimental group ZJO represents only to add vibrio alginolyticus, experimental group 22# represents only to add vibrio alginolyticus Antagonistic Fungi starter, and experimental group ZJO+22# represents to add vibrio alginolyticus and vibrio alginolyticus Antagonistic Fungi starter.
Embodiment:
Following examples are to further illustrate of the present invention, rather than limitation of the present invention.
Embodiment 1: the screening and identification that No. 22, Providence (providencia vermicola)
(1) vibrio alginolyticus Antagonistic Fungi is found, is screened
(A) raw material
Vibrio alginolyticus
Antagonistic strain to be sieved: from marine site, Xisha, lobster enteron aisle, sea cucumber enteron aisle decile be from obtaining 311 strain bacterium.
Used medium reagent: LB substratum, from reagent company, purchase.
LB culture medium prescription: Tryptones 10g/L, yeast extract 5g/L, sodium-chlor 10g/L.
(B) preparation of vibrio alginolyticus liquid: by aseptic technique, transfering loop is inoculated into vibrio alginolyticus in LB substratum, 30 ℃ of 200rpm incubated overnight.
(C) vibrio alginolyticus Antagonistic Fungi separation screening
(1) bacterium is separated
By aseptic technique, lobster enteron aisle in Xisha sampling etc. is ground and is diluted to 10 times of gradients with PBS, respectively get 100ul and coat LA, be placed in 30 ℃ of incubators and cultivate 24h-48h, repeatedly rule 2-3 time, until obtain single bacterium colony, and the cultural characteristic such as size, shape, color, gloss that records bacterium colony in culture dish, is divided into from obtaining 311 strain bacteriums, as Antagonistic Fungi to be sieved, 30 ℃ of 200rpm cultivate 24-48h, as bacterium pure culture liquid to be measured.
(2) screening of Antagonistic Fungi
By aseptic technique, by 10 times of the vibrio alginolyticus dilutions of incubated overnight, get 100ul and coat LA flat board, with punch tool, on substratum, evenly beat afterwards the hole that diameter is 9mm, then draw respectively each 100ul of Antagonistic Fungi fermented liquid to be measured, add in hand-hole, culture dish is cultivated 24-48h in 30 ℃, observe the substratum punching size of inhibition zone around, preliminary screening obtains vibrio alginolyticus to have the bacterial strain of inhibition.
By the bacteriostatic experiment that punches on LA substratum, from 311 strain bacteriums, find a strain to have inhibiting antagonistic strain to vibrio alginolyticus.Code name is No. 22, bacterial strain, its cultural characteristic with to the inhibition of vibrio alginolyticus in Table 1.Its inhibition zone is shown in Fig. 1, negative contrast of Far Left, and middle and the right (arrow indication) is the fungistatic effect figure of Providence bacterium No. 22, and the amount of middle interpolation is less, and restraining effect is little, encloses less.
Table 1: the cultural characteristic that No. 22, bacterial strain and the inhibition to vibrio alginolyticus thereof
(2) Antagonistic Fungi is identified
No. 22, the strain vibrio alginolyticus antagonistic strain screening, its biological property: shaft-like, Gram-negative, wide x is long is (0.6-0.8) x (1.5-2.5) micron.Peritrichous, motion, without natural motility, amphimicrobian.Phenylalanine deamination acid enzyme, Citrate trianion utilization, indole reaction be the positive all, and fermentation seminose produces acid.Arginine dihydrolase, VP reaction be feminine gender all.
Through Automatic Analyzer for Microbes Biolog, identify, again in conjunction with 16SrDNA(as shown in SEQ ID NO.1) sequencing result comparison is known, itself and Providencia vermicola ALK619(KC456588.1) similarity 99%, Providenciavermicola BAB-205(KF535113.1) similarity 99%, Providencia vermicola FFA8(JN092796.1) similarity 99%.Belong to the member in Providence (providencia vermicola), according to qualification result and in conjunction with bacterial strain number, by its called after: No. 22, Providence (providencia vermicola).
No. 22, Providence of the present invention (providencia vermicola), compares with known Providence, all variant aspect form, Physiology and biochemistry and nutritional character.Its principal character is: aerobic, bacterium colony oyster white is regular, opaque.In 4%NaCl, grow, and do not grow in 8%NaCl.Liquefy gelatin, pectin, do not decompose seminose, fructose, maltose, sucrose, lactose.Can decompose semi-lactosi, can utilize glycerine, N.F,USP MANNITOL, arabitol, inositol, but not utilize sorbyl alcohol.Serine, L-glutamic acid, the L-Ala positive, arginine, quininic acid, saccharic acid feminine gender.Can be at pH5 well-grown.
This Providence (providencia vermicola) is preserved in Chinese Typical Representative culture collection center (CCTCC), address No. 22 on December 24th, 2013: Wuhan, China Wuhan University, deposit number is CCTCC No.M2013696.
Embodiment 2: the preparation of Antagonistic Fungi preparation (starter)
The biological property that No. 22, the best vibrio alginolyticus Antagonistic Fungi Providence that obtains according to screening, filters out suitable substratum, and adopts suitable fermentation condition, and this Antagonistic Fungi is made to zymocyte liquid.
The concrete fermentation condition adopting is as follows:
Fermentation strain: No. 22, vibrio alginolyticus Antagonistic Fungi Providence.
Fermentative medium formula: Tryptones 10g, yeast extract 5g, sodium-chlor 10g, distilled water 1L.
Leavening temperature: 30 ℃
Fermentation period: 48-72h
Fermentation shaking speed: 150-200rpm
According to above-mentioned fermentation condition, fermentation obtains vibrio alginolyticus Antagonistic Fungi starter, and its pH is 8.0-8.5.No. 22 concentration of Providence reach 1x10
8-6x10
9cfu/ml.
Embodiment 3: the experiment of halogen worm application simulation
Take 0.2g halogen worm (shrimps) ovum and put into the aseptic seawater of 500ml, under the condition of 30 ℃ of thermostatic pump gas, the hatching of spending the night.With 12 orifice plates, test, prepare in advance the vibrio alginolyticus Antagonistic Fungi starter that embodiment 2 ferments, the aseptic seawater of 2ml and 30 halogen worms are put in every hole, and experimental group is for only adding vibrio alginolyticus (10
8cfu/ml), only add vibrio alginolyticus Antagonistic Fungi starter (10
8and add vibrio alginolyticus (10 cfu/ml),
8cfu/ml) and vibrio alginolyticus Antagonistic Fungi starter (10
8cfu/ml), control group, for not adding any bacterium, only has halogen worm.Every group arrange 3 parallel, experimental session condition does not change, and observes 4d, statistics is respectively organized mortality ratio.Concrete outcome is as shown in table 2 and Fig. 2.
Table 2: each organizes survival rate
By Fig. 2 and table 2, can be found out the vibrio alginolyticus that Providence of the present invention can be removed in water for No. 22, thereby the harm of prevention vibrio alginolyticus to aquaculture object.
Embodiment 4: the fungistatic effect test that No. 22, Providence (providencia vermicola)
The shaker test of the Antagonistic Fungi of reference example 1, carries out the fungistatic effect test of No. 22, Providence (providencia vermicola), and concrete outcome is as shown in table 3:
Table 3
| Indicator | Inhibition zone (diameter d mm) |
| Bacillus subtilis | 11±0.1 |
| Staphylococcus aureus | - |
| C.violaceum CV026 | 12±0.1 |
| Vibrio alginolyticus(E167) | - |
| Vibrio alginolyticus(A187) | - |
| Aeromonas salmonicida199 | 24±0.1 |
| Vibrio parahaemolyticus13150 | 12±0.2 |
| Vibrio harveyi ATCC044 | 10±0.1 |
| Vibrio harveyi BB170 | - |
| Vibrio anguillarum NB10 | - |
| Vbirio alginolyticus1903 | 17±0.1 |
| Vibrio cholerae006 | 8±0.1 |
As can be seen from the above table, Providence of the present invention (providencia vermicola) all has good antagonistic action to the Vibrio of subtilis, chromobacterium violaceum and Vibrio No. 22.
Claims (5)
1. No. 22, Providence (providencia vermicola), and its deposit number is CCTCC No.M2013696.
2. No. 22, Providence claimed in claim 1 (providencia vermicola) application in preparation antibacterials.
3. application according to claim 2, is characterized in that, described antibacterials are prevention and control subtilis or chromobacterium violaceum preparation.
4. application according to claim 2, is characterized in that, described antibacterials are prevention and control aquatic products vibrios preparation.
5. application according to claim 4, is characterized in that, described prevention and control aquatic products vibrios preparation is prevention and control vibrio alginolyticus or Vibrio parahaemolyticus preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410159344.7A CN103966122B (en) | 2014-04-18 | 2014-04-18 | One strain Providence and the application in preparation antibacterials thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410159344.7A CN103966122B (en) | 2014-04-18 | 2014-04-18 | One strain Providence and the application in preparation antibacterials thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103966122A true CN103966122A (en) | 2014-08-06 |
| CN103966122B CN103966122B (en) | 2016-05-11 |
Family
ID=51236130
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410159344.7A Active CN103966122B (en) | 2014-04-18 | 2014-04-18 | One strain Providence and the application in preparation antibacterials thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103966122B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103525803A (en) * | 2013-10-28 | 2014-01-22 | 江南大学 | Preparation method and application of natural material composite system immobilized bifunctional enzyme used for wine |
| CN104480053A (en) * | 2015-01-12 | 2015-04-01 | 山东泰宝生物科技股份有限公司 | Root-knot nematode biocontrol bacterium, antibiological inoculant and application of biocontrol bacterium and antibiological inoculant |
| CN109157546A (en) * | 2018-07-27 | 2019-01-08 | 江苏省海洋水产研究所 | A kind of compound micro-ecological preparation of the acute Hepatopancreatic necrosis syndrome of prevention and control Penaeus Vannmei and its application |
| CN111961619A (en) * | 2020-08-19 | 2020-11-20 | 天津科技大学 | Vibrio maritima capable of producing alginate lyase with good thermal stability and application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102146342A (en) * | 2011-01-11 | 2011-08-10 | 山东潍坊润丰化工有限公司 | Halophilic bacterial agent and preparation method thereof as well as biological treatment system fixed with bacterial agent and application thereof |
| CN103289940A (en) * | 2013-06-19 | 2013-09-11 | 重庆大学 | Providencia rettgeri strain and application thereof |
-
2014
- 2014-04-18 CN CN201410159344.7A patent/CN103966122B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102146342A (en) * | 2011-01-11 | 2011-08-10 | 山东潍坊润丰化工有限公司 | Halophilic bacterial agent and preparation method thereof as well as biological treatment system fixed with bacterial agent and application thereof |
| CN103289940A (en) * | 2013-06-19 | 2013-09-11 | 重庆大学 | Providencia rettgeri strain and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 王永贤: "水牛简达气单胞菌和拉氏普罗威登斯菌的分离与鉴定", 《云南畜牧兽医》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103525803A (en) * | 2013-10-28 | 2014-01-22 | 江南大学 | Preparation method and application of natural material composite system immobilized bifunctional enzyme used for wine |
| CN104480053A (en) * | 2015-01-12 | 2015-04-01 | 山东泰宝生物科技股份有限公司 | Root-knot nematode biocontrol bacterium, antibiological inoculant and application of biocontrol bacterium and antibiological inoculant |
| CN104480053B (en) * | 2015-01-12 | 2018-07-24 | 山东泰宝生物科技股份有限公司 | One plant of root-knot nematode biocontrol bacteria, biocontrol agent and its application |
| CN109157546A (en) * | 2018-07-27 | 2019-01-08 | 江苏省海洋水产研究所 | A kind of compound micro-ecological preparation of the acute Hepatopancreatic necrosis syndrome of prevention and control Penaeus Vannmei and its application |
| CN111961619A (en) * | 2020-08-19 | 2020-11-20 | 天津科技大学 | Vibrio maritima capable of producing alginate lyase with good thermal stability and application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103966122B (en) | 2016-05-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Dash et al. | Biocontrol of luminous vibriosis in shrimp aquaculture: a review of current approaches and future perspectives | |
| Verschuere et al. | Probiotic bacteria as biological control agents in aquaculture | |
| Irianto et al. | Probiotics in aquaculture | |
| Gupta et al. | Growth performance, feed utilization, digestive enzyme activity, innate immunity and protection against Vibrio harveyi of freshwater prawn, Macrobrachium rosenbergii fed diets supplemented with Bacillus coagulans | |
| CN103320365B (en) | Fish-sourced aeromonas hydrophila disease antagonistic strain and application thereof | |
| CN102154176B (en) | Turbot pathogenic strain and inactivated vaccine for ascites disease | |
| US11851644B2 (en) | Marine bacteria formulation useful in aquaculture | |
| CN106544278A (en) | A kind of pathogenic fungi for Blatta seu periplaneta Biological control | |
| Ottesen et al. | Effects on survival and mucous cell proliferation of Atlantic halibut, Hippoglossus hippoglossus L., larvae following microflora manipulation | |
| CN103966122B (en) | One strain Providence and the application in preparation antibacterials thereof | |
| CN107858302A (en) | A kind of bacillus subtilis 7K and its application | |
| Sohn et al. | Probiotic strains for disease management in hatchery larviculture of the eastern oyster Crassostrea virginica | |
| Mastan et al. | Vibriosis in farm reared white shrimp, Litopenaeus vannamei in Andhra Pradesh-natural occurrence and artificial challenge | |
| CN107988171A (en) | Fragility Vibriophage ValSw4-1 and the bactericidal composition comprising the bacteriophage and its application | |
| CN104774781A (en) | Bacillus subtilis DCU and use thereof | |
| CN103387949B (en) | Scophthalmas maximus fin rot disease antagonistic bacterium and application thereof | |
| CN108048412A (en) | Fragility Vibriophage ValLY-4 and the bactericidal composition comprising the bacteriophage and its application | |
| CN108070570A (en) | Fragility Vibriophage ValDsh-1 and the bactericidal composition comprising the bacteriophage and its application | |
| CN103966121A (en) | Pseudomonas aeruginosa and application of pseudomonas aeruginosa in preparing antibacterial drugs | |
| Maheswaran et al. | Screening and characterization of marine seaweeds and its antimicrobial potential against fish pathogens | |
| CN103966123B (en) | One strain Huo Shi enterobacteria and the application in preparation antibacterials thereof | |
| CN103966120B (en) | Bacillus sp. and application of bacillus sp. in preparing antibacterial drugs | |
| Marwiyah et al. | Total plate count and identification of vibrio in pacific white shrimp (Litophenaeus vannamei) from ponds and in those exposed to immunogenic protein membrane Zoothamnium penaei | |
| KR20180131948A (en) | Pseudomonas extremorientalis strain KACC 81047BP and composition for comprising the same | |
| Novita et al. | Potential of Bacillus sp., as a producer of AHL lactonase and its application as a probiotic for the prevention of Mas in catfish (Clarias gariepinus) |
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 |