CN103815138A - Bdellovibrio bacteriovorus nutrient enhancer for holothurian - Google Patents
Bdellovibrio bacteriovorus nutrient enhancer for holothurian Download PDFInfo
- Publication number
- CN103815138A CN103815138A CN201210465682.4A CN201210465682A CN103815138A CN 103815138 A CN103815138 A CN 103815138A CN 201210465682 A CN201210465682 A CN 201210465682A CN 103815138 A CN103815138 A CN 103815138A
- Authority
- CN
- China
- Prior art keywords
- bacteriophagic bdellovibrio
- parts
- bdellovibrio
- culture medium
- bdellovibrio bacteriovorus
- 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.)
- Pending
Links
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a bdellovibrio bacteriovorus nutrient enhancer for holothurian. The weight ratio of a bdellovibrio bacteriovorus agent to nutrient fodder is 1:(2-10). A preparation method for the bdellovibrio bacteriovorus agent comprises the following operation steps: collecting thalli after bdellovibrio bacteriovorus enrichment and washing are performed; drying the thalli after the thalli are mixed with the nutrient fodder to obtain the bdellovibrio bacteriovorus agent so as to obtain a final product with the water content of 2-5% after the bdellovibrio bacteriovorus agent is mixed with the nutrient fodder. The bdellovibrio bacteriovorus nutrient enhancer for holothurian, which is prepared through the method disclosed by the embodiment of the invention, is suitable for holothurian culture; the treating process is simple and time consumption is less; the preparation cost is lower than that of a conventional method; the powder is long in retention time and free of pollution. The bdellovibrio bacteriovorus nutrient enhancer has important practical significance on holothurian culture on a large scale.
Description
Technical field
The present invention relates generally to marine product disease detection technical field, is specifically related to marine product probio nutrient powder, especially a kind of sea cucumber bacteriophagic Bdellovibrio nutrition enhancer.
Background technology
China marine site is vast, superior natural conditions, and marine resources are very abundant.But the degree developing is at present also very low.This also illustrates that China develops having a high potential of marine resources, and marine site, Liaoning is important living marine resources, fishery resources treasure-house of China, and it abounds with sea cucumber, scallop, mussel, oyster and fish, shrimp and algae etc., drives marine economy development.Strengthen polygon and bilateral ocean development international cooperation, concentrate intensive exploitation marine site and island resource, reasonable disposition advantage marine industries, scientific and efficient exploitation marine resources, actively develop scientific research and test, development and utilization, sea-farming standardization is the important means of reasonable development sea-farming resource, studies and defines culturing marine products standard, reasonable development seawater mudflat aquaculture resource, improving cultivation marine product disease is the key content of current aquaculture.
Culturing marine products is a high risk aquaculture, and traditional small family's culturing marine products has brought huge market and technical risk to raiser.Grow risk and drop to minimumly for cultivation is raised on a household basis, carry out all kinds of aquacultures, comprise that the disease prevention and cure task of all kinds of marine products such as fish, shrimp, crab, sea cucumber, abalone, shellfish, marine alga is very important.Report, the harmful bacterias such as comma bacillus, floating vibrios, vibrio parahaemolytious, vibrio alginolyticus, Vibrio vulnificus, are the arch-criminals who causes marine product various diseases.
World's holothruian cultures are take stichopus japonicus (temperate species) and rough sea cucumber (noble cane) as principal item.Except China, Japan, the main culturing stichopus japonicus of Korea S, other countries are all take rough sea cucumber as main cultivation object.In recent years, along with the minimizing of natural resources and the increase of consumption demand, propagating artificially in some countries of sea cucumber risen gradually.Wherein, China and Japan is main cultivation country, and also there are small-scale aquaculture in India, Indonesia, Vietnam, Philippine, Korea S, Egypt etc.
Stichopus japonicus (Apostichopus japonicus) is on the books more than 20 the planting in beche-de-mer and to be worth the highest, the only temperate species that is distributed in territory, the yellow Bohai Sea of China.Its protein content is high, and carbohydrate is abundant, and lipid content is low, and not containing cholesterol, has very high nutrition and health care and medical value, is considered as important precious marine product, thereby has important market value by people.In recent years, increasing year by year of sea cucumber consumption demand amount caused the excessive exploitation of sea cucumber natural resources and the sharply decline of population quantity in world wide, and therefore, propagating artificially also of sea cucumber grows up thereupon.The thing followed is all kinds of problems that run in holothruian cultures.
Bacteriophagic Bdellovibrio (Bdellovibrio bacteriovorus) is current the most frequently used probio kind.Bacteriophagic Bdellovibrio of the present invention, being called for short Bdellovibrio is that a class is attacked, infected, the bacterial parasite of other microorganisms of cracking, it is a kind of probiotics growing up in recent years, most of aquatic pathogenic bacteria are all had to stronger splitting action, it prevents by cracking bacterium or reduces the development of sea cucumber disease and spread, improve sea cucumber inside and outside environment, promote growth, strengthen sea cucumber immunity.Bacteriophagic Bdellovibrio has the title of natural biological modifying agent or natural biological purification factor, biological antibiotic or living antibiotics.The present invention does not limit the kind of bacteriophagic Bdellovibrio, as long as reference culture can be developed into holothruian cultures nutritional preparation, effectively improves the beneficial flora in sea cucumber body.Can reach antibacterial, immune, somatotrophic multiple effect, sea cucumber healthy aquaculture is had to realistic meaning.
Summary of the invention
From background technology, improve the immunity of stichopus japonicus self, strengthening its premunition is the important channel that solves stichopus japonicus disease problem.This research prevents and treats the ill object of sea cucumber in aquaculture for reaching, avoids using antibiotic in sea cucumber disease control, and sea cucumber of the present invention bacteriophagic Bdellovibrio nutrition enhancer, realize by the following technical solutions: formed by following components in certain proportion configuration:
Bacteriophagic Bdellovibrio agent and nutrient fodder weight ratio are 1:2 ~ 10, wherein,
The preparation method of bacteriophagic Bdellovibrio agent comprises following operating procedure:
(1) increase bacterium: be inoculated into bacteriophagic Bdellovibrio culture medium from bacteriophagic Bdellovibrio reference culture, 27～32 ℃ of ventilations were cultivated after 12 hours, again in the volume ratio inoculation bacteriophagic Bdellovibrio host culture medium with 3-5%, ferment, 27～32 ℃ ventilation cultivate 24～48 hours, to the final concentration of bacteria suspension bacterial content be 10
7~ 10
10cfu/g; Make tunning;
(2) washing: the tunning that step (1) is obtained, with the centrifugal 2 ~ 5min of 4000 ~ 8000rpm revolution, to collect thalline; Mass ratio with 1:50 mixes with PBS buffer solution, fully stirs after 1 ~ 5min, collects mixed liquor;
(3) probio proportioning: by the mixed liquor of collecting; Add soy meal by the mass ratio of 1:1, then mix;
(4) dry: the mixture of above-mentioned steps (3) gained mixes with nutrient fodder, mixture than being 1:2 ~ 10, is dried 24 ~ 36h with nutrient fodder gross weight, and product water content is 2 ~ 5%, makes bacteriophagic Bdellovibrio agent; Wherein:
Described nutrient fodder is: 50～70 parts, yellow meal worm sand; Sargassum powder: 25～40 parts; Sargassum 15～35; 10～15 parts of soybean proteins; Fish meal: 5～10 parts; 2～5 parts of spirulinas; 2～5 parts of Mussel Powders; Chitosan oligomer: 1～4 part; 1～4 part of compound vitamin; 1～4 part, compound many ore deposits.
In above-described all technical schemes, preferably condition is as follows: described bacteriophagic Bdellovibrio culture medium, its compound method is as follows: get 4g peptone, 1g dusty yeast, 1g sodium chloride, 10g glucose, 2mg calcium chloride, 2mg magnesium chloride is dissolved in 100ml water, 105 ℃ of sterilizing 20min; After described culture medium is cooling gradually, regulate the pH to 7.5 of described solution for subsequent use.
In above-described all technical schemes, preferably condition is as follows: described bacteriophagic Bdellovibrio cultivation temperature is 30 ℃.
In above-described all technical schemes, preferably condition is as follows: described bacteriophagic Bdellovibrio Host Strains culture medium is colibacillary LB nutrient solution;
In the present invention, in the said goods preparation method, all do not limit other equivalent conditions of method, because it can be determined according to prior art, do not limit the rank of agents useful for same yet, because it is little on result impact, and can be bought and be obtained by preparation or commercial sources, technical staff can make reference according to adjusting according to listed condition in embodiment.These believe that about the selection of preparation way and method those skilled in the art can be enlightened fully from prior art, and the present invention repeats no more.
Advantage of the present invention is: the nursery pond and the growing pond that are applicable to sea cucumber; Processing procedure is simple, consuming time few; Preparation cost is lower than conventional method, and the pulvis holding time of the present invention is long, pollution-free.
The specific embodiment
Following examples are used for illustrating the present invention, but are not used for limiting the scope of the invention.
Embodiment 1
(1) bacterial classification recovery: be inoculated into bacteriophagic Bdellovibrio culture medium from bacteriophagic Bdellovibrio reference culture, under 30 ℃ of conditions, cultivate 12h.
(2) increase bacterium: be inoculated in bacteriophagic Bdellovibrio host culture medium with 4% volume ratio again, under 30 ℃ of conditions, ferment, cultivate 28h, to the final concentration of bacteria suspension bacterial content be 10
8cfu/g; Make tunning;
Wherein, described bacteriophagic Bdellovibrio culture medium, its compound method is as follows:
Get 4g peptone, 1g dusty yeast, 1g sodium chloride, 10g glucose, 2mg calcium chloride, 2mg magnesium chloride is dissolved in 100ml water, 105 ℃ of sterilizing 20min; After described culture medium is cooling gradually, regulate the pH to 7.5 of described solution for subsequent use.
Described bacteriophagic Bdellovibrio Host Strains culture medium is colibacillary LB nutrient solution;
(3) washing: the tunning that step (2) is obtained, with the centrifugal 3min of 5000rpm revolution, to collect thalline; Mass ratio with 1:50 mixes with PBS buffer solution, fully stirs after 3min, collects mixed liquor;
(4) probio proportioning: by the mixed liquor of collecting; Add soy meal by the mass ratio of 1:1, then mix;
(5) dry: the mixture of above-mentioned steps (3) gained mixes with nutrient fodder, mixture than being 1:80, is dried 28h with nutrient fodder gross weight, and product water content is 3%, makes bacteriophagic Bdellovibrio agent;
Described nutrient fodder is: husky 50 parts of yellow meal worm; Sargassum powder: 40 parts; Sargassum 35; 15 parts of soybean proteins; Fish meal: 10 parts; 5 parts of spirulinas; 5 parts of Mussel Powders; Chitosan oligomer: 4 parts; 1 part of compound vitamin; 4 parts, compound many ore deposits.
Embodiment 2
(1) bacterial classification recovery: be inoculated into bacteriophagic Bdellovibrio culture medium from bacteriophagic Bdellovibrio reference culture, under 30 ℃ of conditions, cultivate 12h;
(2) increase bacterium: be inoculated in bacteriophagic Bdellovibrio host culture medium with 5% volume ratio again, under 30 ℃ of conditions, ferment, cultivate 48h, to the final concentration of bacteria suspension bacterial content be 10
10cfu/g; Make tunning;
Wherein, described bacteriophagic Bdellovibrio culture medium, its compound method is as follows: get 4g peptone, 1g dusty yeast, 1g sodium chloride, 10g glucose, 2mg calcium chloride, 2mg magnesium chloride is dissolved in 100ml water, 105 ℃ of sterilizing 20min; After described culture medium is cooling gradually, regulate the pH to 7.5 of described solution for subsequent use.
Described bacteriophagic Bdellovibrio Host Strains culture medium is colibacillary LB nutrient solution;
(3) washing: the tunning that step (2) is obtained, with the centrifugal 3min of 5000rpm revolution, to collect thalline; After fully mixing with PBS buffer solution with the mass ratio of 1:50, collect mixed liquor;
(4) probio proportioning: by the mixed liquor of collecting; Add soy meal by the mass ratio of 1:1, then mix;
(5) dry: the mixture of above-mentioned steps (3) gained mixes with nutrient fodder, mixture than being 1:70, is dried 30h with nutrient fodder gross weight, and product water content is 5%, makes bacteriophagic Bdellovibrio agent;
Described nutrient fodder is: husky 60 parts of yellow meal worm; Sargassum powder: 30 parts; Sargassum 30; 15 parts of soybean proteins; Fish meal: 7 parts; 4 parts of spirulinas; 4 parts of Mussel Powders; Chitosan oligomer: 2 parts; 2 parts of compound vitamins; 2 parts, compound many ore deposits.
Experimental example 3
(1) bacterial classification recovery: bacteriophagic Bdellovibrio reference culture is inoculated in bacteriophagic Bdellovibrio culture medium, cultivates 12h under 30 ℃ of conditions.
(2) increase bacterium: be inoculated in bacteriophagic Bdellovibrio host culture medium with 5% volume ratio again, under 30 ℃ of conditions, carry out fermented and cultured 28h, to the final concentration of bacteria suspension bacterial content be 10
10cfu/g; Make tunning;
Wherein, described bacteriophagic Bdellovibrio culture medium, its compound method is as follows: get 4g peptone, 1g dusty yeast, 1g sodium chloride, 10g glucose, 2mg calcium chloride, 2mg magnesium chloride is dissolved in 100ml water, 105 ℃ of sterilizing 20min; After described culture medium is cooling gradually, regulate the pH to 7.5 of described solution for subsequent use.
Described bacteriophagic Bdellovibrio Host Strains culture medium is colibacillary LB nutrient solution;
(3) washing: the tunning that step (2) is obtained, with the centrifugal 3min of 6000rpm revolution, to collect thalline; Mass ratio with 1:50 mixes with PBS buffer solution, fully stirs after 5min, collects mixed liquor;
(4) probio proportioning: by the mixed liquor of collecting; Add soy meal by the mass ratio of 1:1, then mix;
(5) dry: the mixture of above-mentioned steps (3) gained mixes with nutrient fodder, mixture than being 1:60, is dried 36h with nutrient fodder gross weight, and product water content is 5%, makes bacteriophagic Bdellovibrio agent;
Described nutrient fodder is: husky 50 parts of yellow meal worm; Sargassum powder: 30 parts; Sargassum 15; 15 parts of soybean proteins; Fish meal: 8 parts; 5 parts of spirulinas; 4 parts of Mussel Powders; Chitosan oligomer: 2 parts; 2 parts of compound vitamins; 2 parts, compound many ore deposits.
Experimental example 4
(1) bacterial classification recovery: be inoculated into bacteriophagic Bdellovibrio culture medium from bacteriophagic Bdellovibrio reference culture, cultivate 12h under 30 ℃ of conditions.
(2) increase bacterium: be inoculated in bacteriophagic Bdellovibrio host culture medium with 5% volume ratio again, under 30 ℃ of conditions, ferment, cultivate 48h, to the final concentration of bacteria suspension bacterial content be 10
10cfu/g; Make tunning;
Wherein, described bacteriophagic Bdellovibrio culture medium, its compound method is as follows: get 4g peptone, 1g dusty yeast, 1g sodium chloride, 10g glucose, 2mg calcium chloride, 2mg magnesium chloride is dissolved in 100ml water, 105 ℃ of sterilizing 20min; After described culture medium is cooling gradually, regulate the pH to 7.5 of described solution for subsequent use.
Described bacteriophagic Bdellovibrio Host Strains culture medium is colibacillary LB nutrient solution;
(3) washing: the tunning that step (2) is obtained, with the centrifugal 5min of 4000rpm revolution, to collect thalline; Mass ratio with 1:50 mixes with PBS buffer solution, fully stirs after 5min, collects mixed liquor;
(4) probio proportioning: by the mixed liquor of collecting; Add soy meal by the mass ratio of 1:1, then mix;
(5) dry: the mixture of above-mentioned steps (3) gained mixes with nutrient fodder, mixture than being 1:50, is dried 36h with nutrient fodder gross weight, and product water content is 2%, makes bacteriophagic Bdellovibrio agent;
Described nutrient fodder is: husky 70 parts of yellow meal worm; Sargassum powder: 25 parts; Sargassum 35; 15 parts of soybean proteins; Fish meal: 8 parts; 5 parts of spirulinas; 2 parts of Mussel Powders; Chitosan oligomer: 4 parts; 4 parts of compound vitamins; 4 parts, compound many ore deposits.
Compared with control group sea cucumber, utilize the bacteriophagic Bdellovibrio agent of method preparation described in the embodiment of the present invention 1 ~ 4 to feed sea cucumber, body weight all increases to some extent, and is difficult for ill; Product of the present invention is applicable to holothruian cultures; Processing procedure is simple, consuming time few; Preparation cost is lower than conventional method, and the pulvis holding time of the present invention is long, pollution-free.The present invention has important practical significance to the cultivation that solves sea cucumber in enormous quantities.
The above is only the preferred embodiment of the present invention, it should be pointed out that for those skilled in the art, is not departing under the prerequisite of the technology of the present invention principle, can also make some improvement and also should be considered as protection scope of the present invention.
Claims (3)
1. a sea cucumber bacteriophagic Bdellovibrio nutrition enhancer, is characterized in that: be 1:2 ~ 10 by bacteriophagic Bdellovibrio agent and nutrient fodder weight ratio, wherein:
The preparation method of bacteriophagic Bdellovibrio agent comprises following operating procedure:
(1) increase bacterium: be inoculated into bacteriophagic Bdellovibrio culture medium from bacteriophagic Bdellovibrio reference culture, 27～32 ℃ of ventilations were cultivated after 12 hours, again in the volume ratio inoculation bacteriophagic Bdellovibrio host culture medium with 3-5%, ferment, 27～32 ℃ ventilation cultivate 24～48 hours, to the final concentration of bacteria suspension bacterial content be 10
7~ 10
10cfu/g; Make tunning;
(2) washing: the tunning that step (1) is obtained, with the centrifugal 2 ~ 5min of 4000 ~ 8000rpm revolution, to collect thalline; Mass ratio with 1:50 mixes with PBS buffer solution, fully stirs after 1 ~ 5min, collects mixed liquor;
(3) probio proportioning: by the mixed liquor of collecting; Add soy meal by the mass ratio of 1:1, then mix;
(4) dry: the mixture of above-mentioned steps (3) gained mixes with nutrient fodder, mixture than being 1:2 ~ 10, is dried 24 ~ 36h with nutrient fodder gross weight, and product water content is 2 ~ 5%, makes bacteriophagic Bdellovibrio agent; Wherein:
Described nutrient fodder is: 50～70 parts, yellow meal worm sand; Sargassum powder: 25～40 parts; Sargassum 15～35; 10～15 parts of soybean proteins; Fish meal: 5～10 parts; 2～5 parts of spirulinas; 2～5 parts of Mussel Powders; Chitosan oligomer: 1～4 part; 1～4 part of compound vitamin; 1～4 part, compound many ore deposits.
2. sea cucumber according to claim 1 bacteriophagic Bdellovibrio nutrition enhancer, it is characterized in that, described bacteriophagic Bdellovibrio culture medium, its compound method is as follows: get 4g peptone, 1g dusty yeast, 1g sodium chloride, 10g glucose, 2mg calcium chloride, 2mg magnesium chloride is dissolved in 100ml water, 105 ℃ of sterilizing 20min; After described culture medium is cooling gradually, regulate the pH to 7.5 of described solution for subsequent use.
3. sea cucumber according to claim 2 bacteriophagic Bdellovibrio nutrition enhancer, is characterized in that, described bacteriophagic Bdellovibrio cultivation temperature is 30 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210465682.4A CN103815138A (en) | 2012-11-16 | 2012-11-16 | Bdellovibrio bacteriovorus nutrient enhancer for holothurian |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210465682.4A CN103815138A (en) | 2012-11-16 | 2012-11-16 | Bdellovibrio bacteriovorus nutrient enhancer for holothurian |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103815138A true CN103815138A (en) | 2014-05-28 |
Family
ID=50750667
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210465682.4A Pending CN103815138A (en) | 2012-11-16 | 2012-11-16 | Bdellovibrio bacteriovorus nutrient enhancer for holothurian |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103815138A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110241057A (en) * | 2019-07-23 | 2019-09-17 | 福建九为生物技术有限公司 | A kind of direct putting type Bdellovibrio leavening and preparation method thereof and application method |
-
2012
- 2012-11-16 CN CN201210465682.4A patent/CN103815138A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110241057A (en) * | 2019-07-23 | 2019-09-17 | 福建九为生物技术有限公司 | A kind of direct putting type Bdellovibrio leavening and preparation method thereof and application method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhou et al. | Role and functions of beneficial microorganisms in sustainable aquaculture | |
| CN101983582B (en) | Traditional Chinese medicine microecological preparation for preventing and treating enteritis of grass carp and preparation method thereof | |
| CN101638627B (en) | Bacillus subtilis and application thereof in biological feed additives | |
| CN103275907B (en) | Bacillus amyloliquefacien and preparation method and application thereof | |
| CN101671638B (en) | New strain of bifidobacterium and fermentative preparation method and application thereof | |
| CN102517227B (en) | Enterococcus faecalis and applications and feed additive and leavening agent thereof | |
| CN102559560A (en) | Bacillus licheniformis for producing lactic acid at high yield after fermentation, and preparation and application of Bacillus licheniformis | |
| CN103815150A (en) | Lactic acid bacterium nutrient enhancer for holothurian culture | |
| CN101891307B (en) | Marine culture biological cleanser and preparation method thereof | |
| CN102669486B (en) | Special animal biogen for shrimps and preparation method thereof | |
| CN102017910B (en) | Energy-saving and emission-reducing culture method of turbot | |
| CN111728081B (en) | Composite bacteria fermentation liquor for feed additive and preparation method thereof | |
| CN103815138A (en) | Bdellovibrio bacteriovorus nutrient enhancer for holothurian | |
| Hu et al. | Synthetic microbiome for a sustainable poultry industry | |
| CN105838640A (en) | Composite probiotics for stichopus japonicus feeding and preparation method thereof | |
| CN102038714A (en) | Application of bdellovibrio bdelloplast in preparing bacterial agent for preventing and controlling red mouth disease of scophthalmus maximus | |
| CN103815139A (en) | Preparation method for bdellovibrio bacteriovorus nutrient enhancer for holothurian | |
| CN103815158A (en) | Preparation method for medical stone vector bdellovibrio bacteriovorus agent for holothurian | |
| CN103815137A (en) | Preparation method for lactic acid bacterium nutrient enhancer for holothurian culture | |
| CN103815162A (en) | Medical stone carried lactic acid bacteria agent for cultivating sea cucumbers | |
| CN103815167A (en) | Anti-vibrio alginnolyficus nutrition enhancer for sea cucumbers | |
| CN103815159A (en) | Bacillus nutrient enhancer for holothurian | |
| CN103815168A (en) | Method for preparing anti-vibrio alginnolyficus nutrition enhancer for sea cucumbers | |
| CN103815170A (en) | Method for preparing bacillus nutrition enhancer for sea cucumbers | |
| CN103815151A (en) | Medical stone vector bdellovibrio bacteriovorus agent for holothurian |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140528 |