CN104974949A - Screening method of insecticidal bacteria aiming to marine fish philasterides dicentrarchi antigen - Google Patents
Screening method of insecticidal bacteria aiming to marine fish philasterides dicentrarchi antigen Download PDFInfo
- Publication number
- CN104974949A CN104974949A CN201410149540.6A CN201410149540A CN104974949A CN 104974949 A CN104974949 A CN 104974949A CN 201410149540 A CN201410149540 A CN 201410149540A CN 104974949 A CN104974949 A CN 104974949A
- Authority
- CN
- China
- Prior art keywords
- seawater
- fish
- shield
- bacteria
- bacterium
- 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
Abstract
The invention provides a screening method of insecticidal bacteria aiming to marine fish philasterides dicentrarchi antigen. The screening method includes following steps: (1) separating and purifying marine bacteria; (2) separating and screening the marine fish philasterides dicentrarchi antigen; (3) performing co-cultivation to the separated marine bacteria with philasterides dicentrarchi in a 96-well plate; and (4) monitoring the survival rate of philasterides dicentrarchi in the co-cultivation system to determine the insecticidal bacteria. The screening method is advantaged in that on the basis of the researching works in the prior art, the method can screen the marine bacteria in a large scale rapidly and can obtain the bacterial strain which has a significant effect and has a definite insecticidal effect on target philasterides dicentrarchi, thereby increasing the possibility of preventing and treating the philasterides dicentrarchi diseases with a microorganism method. The screening method is less in required time, is increased in efficiency, is free of pollution in the whole experiment process and can achieve a green and environment-friendly demand.
Description
Technical field
The invention belongs to microorganism field, particularly relate to a kind of screening method for seawater fish shield ciliate cause of disease insecticidal bacteria.
Background technology
Sea water fish industrialized culture is a very important industry in China's aquaculture, but along with the continuous expansion improving constantly and cultivate scale of cultured output, the generation of various disease is also further frequent, and wherein shield balantidiasis wherein endangers larger one.Shield cilium Eimeria facultative parasite, generally seeks free living, with the fine particulate matter suspended (bacterium, micro-algae, protozoon etc.) for food; But in some environments, these ciliates can show as parasitics, engulf fish biological cells and tissues residue and in its tissue in-growth, breeding; Report display, all can there is shield balantidiasis from 12 ~ 26 DEG C in industrialized culture workshop, China various places, almost all can fall ill the whole year; Shield balantidiasis is the most violent in lefteye flounder morbidity, control ineffective there will be more than 90% mortality ratio.The fine class ciliate of the shield found in current China sea water fish industrialized culture mainly contains Parauronema virginianum, Mesanophrys carcini, Paralembus digitiformis, the fine worm of the pseudo-health of water droplet, voracious Miami worm.
For prevention and control shield balantidiasis is to the harm of China's marine fish culture industry, domestic Duo Jia unit has carried out the screening of protective agents for shield ciliate, but at present aquatic products still concentrates on formalin to the treatment of this disease, copper sulfate, the use of zinc sulfate, these medicines act on protein denaturation substantially, although it can kill shield ciliate in water body, but be difficult to radical cure and kill the shield ciliate in fish body, and these medicines do not have distinguishing ability to fish and human body cell, can play a role equally, lead to grave consequences, Chinese patent literature CN1762460A in addition, CN103349747A discloses the Chinese medicine preparation of the control lefteye flounder shield balantidiasis of two kinds of Chinese herbal medicine formulas, but the application of such preparation in actual production and stability also need more multiple testing.
In fact, at nature, also exist many have shield ciliate cause a disease or the microorganism of killing action, utilize this pathogenic shield balantidiasis of preventing and treating of microorganism can be a kind of effective biological control method, the method belongs to microbial control insect field, its research started from for 19th century, the exploitation practical stage was progressed into the upper semiduation in 20th century, microbial pesticide is that bio-science is combined the product developed with engineering, along with developing rapidly of modern biological project, microbial pesticide will have very large DEVELOPMENT PROSPECT, but according to current data, also do not find the method report carrying out insecticidal bacteria screening for seawater fish shield ciliate cause of disease at present.
Summary of the invention
The object of the invention is the defect for prior art, provides a kind of screening method for seawater fish shield ciliate cause of disease insecticidal bacteria, can efficient, in enormous quantities, the rapid screening carrying out insecticidal bacteria for seawater fish shield ciliate cause of disease by the method.
The present invention is achieved by the following technical solutions, have developed a kind of screening method for seawater fish shield ciliate cause of disease insecticidal bacteria, it is characterized in that, comprise the following steps:
(1), separation and purification obtains marine bacteria: get habitat, ocean source sample, after adding sterilizing seawater concussion mixing, be configured to 10
-2, 10
-3, 10
-4, 10
-5diluent, be coated with on 100 μ l to improvement seawater 2216E solid medium flat board respectively with above-mentioned dilution diluent, each dilution repetition twice, be inverted in 25 DEG C with coated culture dish and cultivate 24-96 hour, single bacterium colony that picking grows, rule at seawater 2216E solid medium flat board, the culture dish pulling line is inverted in 25 DEG C and cultivates 24-96 hour, single bacterium colony that picking grows, again rule on seawater 2216E solid medium flat board, the culture dish pulling line is inverted in 25 DEG C and cultivates 24-96 hour, treat that single bacterium colony grows, picking list bacterium colony is in 1ml seawater 2216E liquid nutrient medium, and it is for subsequent use with sealed membrane sealing, the seawater 2216E liquid nutrient medium of the good single bacterium colony of inoculation is placed in 25 DEG C and shakes bacterium 24-96 hour, 0.4-0.7 is reached to its OD600, seal with sealed membrane, for subsequent use in Bacteria liquid,
(2), separation screening seawater fish shield ciliate cause of disease: marine fish culture factory is fetched the sea water fish of suffering from shield balantidiasis, get its gill portion, brain, body surface focus, make water logging sheet microscopic examination lesions position, get and find that there is the ciliophoran lesions position of shield and join in 5 milliliters of sterilizing seawater and shake, be configured to 10
-2, 10
-3, 10
-4, 10
-5diluent, above-mentioned dilution diluent is got 10 μ l respectively to be dripped on sterilized slide glass, by the ciliophoran quantity of shield in each water droplet of microscopic examination, an each existence ciliophoran water droplet of shield is transferred in 1ml fish soup substratum immediately and is placed in 14-18 DEG C of cultivation, treat that in fish soup substratum, shield ciliate density reaches 5 × 10
3individual/more than ml time, by be placed in this 1ml fish soup media transfer to 50ml fish soup substratum 14-18 DEG C continue cultivate, until shield ciliate density reaches 5 × 10 in this 50ml fish soup substratum
3individual/more than ml, for subsequent use as shield ciliate nutrient solution;
(3), the marine bacteria and shield ciliate that are separated acquisition are carried out Dual culture in 96 orifice plates: get 96 orifice plates, blank is done in A1, B1 hole of 96 orifice plates, and add 160 μ l sterilizing seawater, all the other holes are experimental group; Getting step (1) Bacteria liquid 160 μ l joins in 96 orifice plates, often kind of bacterium establish 2 parallel, namely each bacterium adds holes; Get the shield ciliate nutrient solution that step (2) obtains, to annotate 160 μ l shield ciliate nutrient solutions to the annotated Kong Zhongzai of sterilizing seawater or Bacteria liquid of 96 orifice plates successively, excellent 96 orifice plates will be added and be placed in wet box, and be put into 16 DEG C of Dual culture; (4), shield ciliophoran survival rate determination insecticidal bacteria in co-culture system is monitored: after starting Dual culture, every 1 day, 96 orifice plates are taken out from incubator, every hole liquid is blown even rear taking-up 10 μ l-20 μ l liquid and examine under a microscope shield ciliate growing state and survival rate, observe active shield ciliate individuality in liquid and be less than 10, this bacterium is effective bacterium, and effective bacterium preservation is retained.
Basigamy method cultivated by fish soup in described step (2): get the old seawater of 50ml, through 121 DEG C, 20min sterilizing, adds 1ml fish bouillon mother liquor after cooling.
In described step (4), effective bacterium preservation is retained, and its method is, is saved backup by effective bacterium plate is placed in 25 DEG C in picking list bacterium colony to 1 milliliter of seawater 2216E liquid nutrient medium shakes bacterium 36-96 hour, to its OD
600reach 0.4-0.6, add the glycerine of sterilizing, make the final concentration of glycerine be 30-50% ,-80 DEG C save backup.
Described fish bouillon mother liquor collocation method is: get turbot flesh of fish 10-30g in Erlenmeyer flask, add the distilled water of 50ml, electric furnace boils 10min cooling, is divided by supernatant liquor in the centrifuge tube of the 1.5ml being filled to bacterium of having gone out, often pipe packing 1ml, is positioned over-20 DEG C of refrigerators for subsequent use.
Habitat, ocean source sample in described step (1) mainly comprises seawater, marine bottom sediment and marine organisms.
Positively effect of the present invention: on the basis of former research, pass through the present invention, can in enormous quantities, carry out rapidly the screening of marine bacteria, and obtain action effect has clear and definite killing action significantly bacterial strain to object shield ciliate, thus improve the possibility of using microbe method control shield balantidiasis, have consuming time short, raise the efficiency, pollution-free in whole experimentation, the requirement of environmental protection can be reached.
Embodiment
Embodiment 1
Substratum collocation method:
Seawater 2216E solid culture based formulas is: containing peptone 5g, yeast extract paste 1g, tertiary iron phosphate 0.01g, agar 20g in every 1 liter of Chen Haishui;
Seawater 2216E liquid culture based formulas is: containing peptone 5g, yeast extract paste 1g, tertiary iron phosphate 0.01g in every 1 liter of Chen Haishui;
Fish bouillon mother liquor collocation method is: get turbot flesh of fish 20g in Erlenmeyer flask, add the distilled water of 50ml, electric furnace boils 10min, cooling; Supernatant liquor is divided in the centrifuge tube of the 1.5ml being filled to bacterium of having gone out, often pipe packing 1ml, be positioned over-20 DEG C of refrigerators for subsequent use;
Fish soup substratum collocation method is: get the old seawater of 50ml, and through 121 DEG C, 20min sterilizing, adds 1ml fish bouillon mother liquor after cooling.
Embodiment 2
The bacterium having insecticidal effect to seawater fish shield ciliate cause of disease is separated from the turbot cultivating pool pond water sample gathered:
1), separation and purification obtains marine bacteria: get 1 milliliter of turbot cultivating pool Chi Shui, after adding sterilizing seawater concussion mixing, be configured to 10
-2, 10
-3, 10
-4, 10
-5diluent; Different dilution diluent is coated with on 100 μ l to seawater 2216E solid medium flat board respectively, each dilution repetition twice; Coated culture dish is inverted in 25 DEG C cultivate 38 hours, single bacterium colony that picking grows, rules at seawater 2216E solid medium flat board; The culture dish pulling line is inverted in 25 DEG C to cultivate 38 hours, single bacterium colony that picking grows, rules again on seawater 2216E solid medium flat board; The culture dish pulling line is inverted in 25 DEG C to cultivate 38 hours, treats that single bacterium colony grows; Picking list bacterium colony in 1ml seawater 2216E liquid nutrient medium, and seals for subsequent use with sealed membrane; The seawater 2216E liquid nutrient medium of the good single bacterium colony of inoculation is placed in 25 DEG C and shakes bacterium more than 38 hours, to its OD
600reach 0.4, with sealed membrane sealing, this Bacteria liquid is for subsequent use;
2), separation screening seawater fish shield ciliate cause of disease: marine fish culture factory is fetched the sea water fish of suffering from shield balantidiasis, get its gill portion, brain, body surface focus, make water logging sheet microscopic examination lesions position, get and find that there is the ciliophoran lesions position of shield and join in 5 milliliters of sterilizing seawater and shake, be configured to 10
-2, 10
-3, 10
-4, 10
-5diluent, dilution for difference diluent is got 10 μ l respectively to be dripped on sterilized slide glass, by the ciliophoran quantity of shield in each water droplet of microscopic examination, an each existence ciliophoran water droplet of shield is transferred in 1ml fish soup substratum immediately and is placed in 16 DEG C of cultivations, treat that in fish soup substratum, shield ciliate density reaches 5 × 10
3individual/more than ml time, by be placed in this 1ml fish soup media transfer to 50ml fish soup substratum 16 DEG C continue cultivate, until shield ciliate density reaches 5 × 10 in this 50ml fish soup substratum
3individual/ml, for subsequent use in shield ciliate nutrient solution;
3), the marine bacteria and shield ciliate that are separated acquisition are carried out Dual culture in 96 orifice plates: get 96 orifice plates, blank is done in A1, B1 hole of 96 orifice plates, and add 160 μ l sterilizing seawater, all the other holes are experimental group; Getting step (1) Bacteria liquid 160 μ l joins in 96 orifice plates, often kind of bacterium establish 2 parallel, namely each bacterium adds holes; Get the shield ciliate nutrient solution that step (2) obtains, to annotate 160 μ l shield ciliate nutrient solutions to the annotated Kong Zhongzai of sterilizing seawater or Bacteria liquid of 96 orifice plates successively, excellent 96 orifice plates will be added and be placed in wet box, and be put into 16 DEG C of Dual culture;
4), shield ciliophoran survival rate determination insecticidal bacteria in co-culture system is monitored: after starting Dual culture, every 1 day, 96 orifice plates are taken out from incubator, every hole liquid is blown even rear taking-up 10 μ l liquid and examine under a microscope shield ciliate growing state and survival rate, if when not observing active shield ciliate individuality in the liquid that certain hole is taken out, again get 50 μ l liquid from respective aperture to observe, if observe active shield ciliate individuality in liquid to be less than 10, and kindred circumstances appears in Duplicate Samples, then judge that its preservation to be retained as effective bacterium by this bacterium.
Embodiment 3
The bacterium having insecticidal effect to seawater fish shield ciliate cause of disease is separated sea cucumber culture pond bed mud sample from the sunshine gathered:
1), separation and purification obtains marine bacteria: get 1g sea cucumber culture pond at sunshine bed mud sample, after adding 10ml sterilizing seawater concussion mixing, be configured to 10
-2, 10
-3, 10
-4, 10
-5diluent; Different dilution diluent is coated with on 100 μ l to seawater 2216E solid medium flat board respectively, each dilution repetition twice; Coated culture dish is inverted in 25 DEG C cultivate 40 hours, single bacterium colony that picking grows, rules at seawater 2216E solid medium flat board; The culture dish pulling line is inverted in 25 DEG C to cultivate 40 hours, single bacterium colony that picking grows, rules again on seawater 2216E solid medium flat board; The culture dish pulling line is inverted in 25 DEG C to cultivate more than 40 hours, treats that single bacterium colony grows; Picking list bacterium colony is in 1m1 seawater 2216E liquid nutrient medium, and the sealing of this plate sealed membrane is for subsequent use; The seawater 2216E liquid nutrient medium of the good single bacterium colony of inoculation is placed in 25 DEG C and shakes bacterium 40 hours, to its 0D
600reach more than 0.6, with sealed membrane sealing, this Bacteria liquid is for subsequent use;
2), separation screening seawater fish shield ciliate cause of disease: fetch the sea water fish of suffering from shield balantidiasis from marine fish culture factory, get its gill portion, brain, body surface focus, make water logging sheet microscopic examination lesions position; Get and find that there is the ciliophoran lesions position of shield and join in 5 milliliters of sterilizing seawater and shake, be configured to 10
-2, 10
-3, 10
-4, 10
-5diluent; Dilution for difference diluent is got 10 μ l respectively to be dripped on sterilized slide glass, by the ciliophoran quantity of shield in each water droplet of microscopic examination, transfers to immediately in 1ml fish soup substratum be placed in 16 DEG C of cultivations to an each existence ciliophoran water droplet of shield; Treat that in fish soup substratum, shield ciliate density reaches 5 × 10
3during/more than ml, this 1ml fish soup substratum is all transferred in 50ml fish soup substratum and is placed in 16 DEG C of continuation cultivations, until shield ciliate density reaches 5 × 10 in this 50ml fish soup substratum
3/ more than ml, for subsequent use in shield ciliate nutrient solution;
3), the marine bacteria and shield ciliate that are separated acquisition are carried out Dual culture in 96 orifice plates: get 96 orifice plates, blank is done in A1, B1 hole of 96 orifice plates, and add 160 μ l sterilizing seawater, all the other holes are experimental group; Get step 1) Bacteria liquid 160 μ l joins in 96 orifice plates, often kind of bacterium establish 2 parallel, namely each bacterium adds holes; Get step 2) the shield ciliate nutrient solution that obtains, to annotate 160 μ l shield ciliate nutrient solutions to the annotated Kong Zhongzai of sterilizing seawater or Bacteria liquid of 96 orifice plates successively; Excellent 96 orifice plates will be added and be placed in wet box, be put into 16 DEG C of Dual culture;
4), shield ciliophoran survival rate determination insecticidal bacteria in co-culture system is monitored: after starting Dual culture, every 1 day, 96 orifice plates are taken out from incubator, every hole liquid is blown even rear taking-up 20 μ l liquid and examine under a microscope shield ciliate growing state and survival rate, if when not observing active shield ciliate individuality in the liquid that certain hole is taken out, again get 100 μ l liquid from respective aperture to observe, if observe active shield ciliate individuality in liquid to be less than 10, and kindred circumstances appears in Duplicate Samples, then judge that its preservation to be retained as effective bacterium by this bacterium.
Below the turbot cultivating pool pond water sample only gathered and sunshine are separated the bacterium to the original insecticidal effect of seawater fish shield balantidiasis in sea cucumber culture pond bed mud sample, screening method for seawater fish shield ciliate cause of disease insecticidal bacteria is described, do not limit by embodiment during concrete enforcement, without departing from the present invention, various change and modification can also be made, institute's operation technique term in present method, except as otherwise noted, generally there is the implication that those of ordinary skill in the art understand usually, therefore all equivalent technical schemes also should belong to category of the present invention.
Claims (5)
1., for a screening method for seawater fish shield ciliate cause of disease insecticidal bacteria, it is characterized in that, comprise the following steps:
(1), separation and purification obtains marine bacteria: get habitat, ocean source sample, after adding sterilizing seawater concussion mixing, be configured to 10
-2, 10
-3, 10
-4, 10
-5diluent, be coated with on 100 μ l to improvement seawater 2216E solid medium flat board respectively with above-mentioned dilution diluent, each dilution repetition twice, be inverted in 25 DEG C with coated culture dish and cultivate 24-96 hour, single bacterium colony that picking grows, rule at seawater 2216E solid medium flat board, the culture dish pulling line is inverted in 25 DEG C and cultivates 24-96 hour, single bacterium colony that picking grows, again rule on seawater 2216E solid medium flat board, the culture dish pulling line is inverted in 25 DEG C and cultivates 24-96 hour, treat that single bacterium colony grows, picking list bacterium colony is in 1ml seawater 2216E liquid nutrient medium, and it is for subsequent use with sealed membrane sealing, the seawater 2216E liquid nutrient medium of the good single bacterium colony of inoculation is placed in 25 DEG C and shakes bacterium 24-96 hour, 0.4-0.7 is reached to its OD600, seal with sealed membrane, for subsequent use in Bacteria liquid,
(2), separation screening seawater fish shield ciliate cause of disease: marine fish culture factory is fetched the sea water fish of suffering from shield balantidiasis, get its gill portion, brain, body surface focus, make water logging sheet microscopic examination lesions position, get and find that there is the ciliophoran lesions position of shield and join in 5 milliliters of sterilizing seawater and shake, be configured to 10
-2, 10
-3, 10
-4, 10
-5diluent, above-mentioned dilution diluent is got 10 μ l respectively to be dripped on sterilized slide glass, by the ciliophoran quantity of shield in each water droplet of microscopic examination, an each existence ciliophoran water droplet of shield is transferred in 1ml fish soup substratum immediately and is placed in 14-18 DEG C of cultivation, treat that in fish soup substratum, shield ciliate density reaches 5 × 10
3individual/more than ml time, by be placed in this 1ml fish soup media transfer to 50ml fish soup substratum 14-18 DEG C continue cultivate, until shield ciliate density reaches 5 × 10 in this 50ml fish soup substratum
3individual/more than ml, for subsequent use as shield ciliate nutrient solution;
(3), the marine bacteria and shield ciliate that are separated acquisition are carried out Dual culture in 96 orifice plates: get 96 orifice plates, blank is done in A1, B1 hole of 96 orifice plates, and add 160 μ l sterilizing seawater, all the other holes are experimental group; Getting step (1) Bacteria liquid 160 μ l joins in 96 orifice plates, often kind of bacterium establish 2 parallel, namely each bacterium adds holes; Get the shield ciliate nutrient solution that step (2) obtains, to annotate 160 μ l shield ciliate nutrient solutions to the annotated Kong Zhongzai of sterilizing seawater or Bacteria liquid of 96 orifice plates successively, excellent 96 orifice plates will be added and be placed in wet box, and be put into 16 DEG C of Dual culture;
(4), shield ciliophoran survival rate determination insecticidal bacteria in co-culture system is monitored: after starting Dual culture, every 1 day, 96 orifice plates are taken out from incubator, every hole liquid is blown even rear taking-up 10 μ l-20 μ l liquid and examine under a microscope shield ciliate growing state and survival rate, observe active shield ciliate individuality in liquid and be less than 10, this bacterium is effective bacterium, and effective bacterium preservation is retained.
2. a kind of screening method for seawater fish shield ciliate cause of disease insecticidal bacteria according to claim l, it is characterized in that: the fish soup in described step (2) is cultivated basigamy method and is: get the old seawater of 50ml, through 121 DEG C, 20min sterilizing, adds 1ml fish bouillon mother liquor after cooling.
3. a kind of screening method for seawater fish shield ciliate cause of disease insecticidal bacteria according to claim 1, it is characterized in that: in described step (4), effective bacterium preservation is retained, its method is, effective bacterium is saved backup and plate is placed in 25 DEG C in picking list bacterium colony to 1 milliliter of seawater 2216E liquid nutrient medium shakes bacterium 36-96 hour, to its OD
600reach 0.4-0.6, add the glycerine of sterilizing, make the final concentration of glycerine be 30-50% ,-80 DEG C save backup.
4. a kind of screening method for seawater fish shield ciliate cause of disease insecticidal bacteria according to claim 2, it is characterized in that: described fish bouillon mother liquor collocation method is: get turbot flesh of fish 10-30g in Erlenmeyer flask, add the distilled water of 50ml, electric furnace boils 10min cooling, supernatant liquor is divided in the centrifuge tube of the 1.5ml being filled to bacterium of having gone out, often pipe packing 1ml, is positioned over-20 DEG C of refrigerators for subsequent use.
5. a kind of screening method for seawater fish shield ciliate cause of disease insecticidal bacteria according to claim 1, is characterized in that: habitat, the ocean source sample in described step (1) mainly comprises seawater, marine bottom sediment and marine organisms.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410149540.6A CN104974949B (en) | 2014-04-08 | 2014-04-08 | A kind of screening technique for seawater fish shield infusorian cause of disease insecticidal bacteria |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410149540.6A CN104974949B (en) | 2014-04-08 | 2014-04-08 | A kind of screening technique for seawater fish shield infusorian cause of disease insecticidal bacteria |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104974949A true CN104974949A (en) | 2015-10-14 |
| CN104974949B CN104974949B (en) | 2018-02-13 |
Family
ID=54271916
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410149540.6A Expired - Fee Related CN104974949B (en) | 2014-04-08 | 2014-04-08 | A kind of screening technique for seawater fish shield infusorian cause of disease insecticidal bacteria |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104974949B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105199999A (en) * | 2015-11-15 | 2015-12-30 | 国家海洋局第一海洋研究所 | Zunongwangia profunda and application thereof |
| CN105624075A (en) * | 2016-04-05 | 2016-06-01 | 国家海洋局第一海洋研究所 | Salinivibrio costicola strain |
| CN111154652A (en) * | 2020-01-08 | 2020-05-15 | 华南师范大学 | Free-living ciliate protozoan culture medium and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1762460A (en) * | 2005-09-23 | 2006-04-26 | 中国水产科学研究院黄海水产研究所 | The Chinese herbal medicine compound prescription of control seawater fish shield balantidiasis |
| CN102559509A (en) * | 2011-11-07 | 2012-07-11 | 天津师范大学 | Screening and application of Bohai Bay acremonium cephalosporium |
| CN103387949A (en) * | 2013-08-04 | 2013-11-13 | 国家海洋局第一海洋研究所 | Scophthalmas maximus fin rot disease antagonistic bacterium and application thereof |
-
2014
- 2014-04-08 CN CN201410149540.6A patent/CN104974949B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1762460A (en) * | 2005-09-23 | 2006-04-26 | 中国水产科学研究院黄海水产研究所 | The Chinese herbal medicine compound prescription of control seawater fish shield balantidiasis |
| CN102559509A (en) * | 2011-11-07 | 2012-07-11 | 天津师范大学 | Screening and application of Bohai Bay acremonium cephalosporium |
| CN103387949A (en) * | 2013-08-04 | 2013-11-13 | 国家海洋局第一海洋研究所 | Scophthalmas maximus fin rot disease antagonistic bacterium and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| L.LEON-RODRIGUEZ ET AL.: "A vaccine based on biodegradable microspheres induces protective against scuticociliatosis without producing side effects in turbot", 《FISH & SHELLFISH IMMUNOLOGY》 * |
| 胡志钰等: "海洋放线菌杀虫抗生素的一种快速筛选模型", 《海洋通报》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105199999A (en) * | 2015-11-15 | 2015-12-30 | 国家海洋局第一海洋研究所 | Zunongwangia profunda and application thereof |
| CN105199999B (en) * | 2015-11-15 | 2018-02-27 | 国家海洋局第一海洋研究所 | A kind of deep layer Wang Zunong Salmonella and its application |
| CN105624075A (en) * | 2016-04-05 | 2016-06-01 | 国家海洋局第一海洋研究所 | Salinivibrio costicola strain |
| CN105624075B (en) * | 2016-04-05 | 2019-04-09 | 国家海洋局第一海洋研究所 | A kind of raw salt vibrios of rib |
| CN111154652A (en) * | 2020-01-08 | 2020-05-15 | 华南师范大学 | Free-living ciliate protozoan culture medium and application thereof |
| CN111154652B (en) * | 2020-01-08 | 2020-11-27 | 华南师范大学 | Free-living ciliate protozoan culture medium and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104974949B (en) | 2018-02-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108676756B (en) | Bei Laisi bacillus and its application as aquatic pathogenic bacterium inhibitor | |
| CN103952348B (en) | Brevibacterium frigoritolerans as well as microbial inoculant and application of brevibacterium frigoritolerans and microbial inoculant | |
| CN106544278A (en) | A kind of pathogenic fungi for Blatta seu periplaneta Biological control | |
| CN102994417B (en) | Bacillus pumilus and application thereof in prevention and control of comma bacillus in aquatic product | |
| CN104974949A (en) | Screening method of insecticidal bacteria aiming to marine fish philasterides dicentrarchi antigen | |
| CN104830728B (en) | A kind of providencia rettgeri | |
| CN104403958B (en) | One plant can effectively suppress the bacillus cereus NY5 of Rofe source of fish Streptococcusagalactiae | |
| CN106987539B (en) | A kind of 6262 fermentation liquid of bacterium Lysobacter soli and its application | |
| CN106148264B (en) | One plant of bacillus amyloliquefaciens with the molten algae ability enhanced and its application | |
| CN108841748A (en) | Sinorhizobium nitrogen-fixing bacteria strain H6 and its application | |
| Zhang et al. | Bioremediation of bacteria pollution using the marine sponge Hymeniacidon perlevis in the intensive mariculture water system of turbot Scophthalmus maximus | |
| CN107674850A (en) | Certain kind of berries reality pseudomonas Sned811, metabolite and the application of a kind of killing root-knot nematode | |
| CN105695376B (en) | One plant of simple bacillus for killing plant root-knot nematode and its application | |
| CN114437964B (en) | Bacillus belicus strain and application thereof | |
| CN109609405A (en) | Produce bacillus and the purposes of algistatic activity substance | |
| CN108102943A (en) | A kind of efficient denitrification microorganism and its application | |
| CN105199999B (en) | A kind of deep layer Wang Zunong Salmonella and its application | |
| CN105176887A (en) | Method for culturing escherichia coli | |
| CN107012104B (en) | Bacillus subtilis, microecological preparation, feed additive, premix and feed | |
| CN105624075B (en) | A kind of raw salt vibrios of rib | |
| CN1807570A (en) | Process for industrialized production of microsporidian using silkworm as host | |
| CN113980815B (en) | Chlorella with characteristic of guangsalt and application thereof | |
| CN103966120B (en) | Bacillus sp. and application of bacillus sp. in preparing antibacterial drugs | |
| CN110396482B (en) | Antagonistic bacterium bacillus amyloliquefaciens X8 strain and application thereof | |
| Hossain et al. | Aspergillus niger grows faster than Escherichia coli in eosin methylene blue media and deter their growth by reducing the pH of the media |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180213 Termination date: 20200408 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |