CN101919408B - Application of photosynthetic bacterium and bacillus subtilis mixed bacterial liquid in preparing microbial inoculum for preventing and controlling red mouth disease of turbot - Google Patents
Application of photosynthetic bacterium and bacillus subtilis mixed bacterial liquid in preparing microbial inoculum for preventing and controlling red mouth disease of turbot Download PDFInfo
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- CN101919408B CN101919408B CN2010102684356A CN201010268435A CN101919408B CN 101919408 B CN101919408 B CN 101919408B CN 2010102684356 A CN2010102684356 A CN 2010102684356A CN 201010268435 A CN201010268435 A CN 201010268435A CN 101919408 B CN101919408 B CN 101919408B
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- bacillus subtilis
- photosynthetic bacterium
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Abstract
The invention discloses an application of photosynthetic bacterium and bacillus subtilis mixed bacterial liquid in preparing microbial inoculum for preventing and controlling red mouth disease of turbot. The application is characterized in that photosynthetic bacterium culture medium and bacillus subtilis culture medium are mixed according to the ratio of 1:1 on the basis of the number of bacteria; the photosynthetic bacterium is at least one of rhodopseudomonas, rhodopseudomonas palustris and capsule erythrocyte. The application method of the bacterial liquid comprises the following steps: spraying bactericide into a water body to ensure that the final concentration of the photosynthetic bacterium and bacillus subtilis mixed bacterial liquid is between 10 and 107cfu/ml, or dipping feed in the photosynthetic bacterium and bacillus subtilis mixed bacterial liquid, wherein the concentration of the photosynthetic bacterium and the bacillus subtilis in the mixed bacterial liquid is between 10 and 107cfu/mL. The bactericide is effective, environement-friendly and without public nuisance, has excellent application prospect, and has important guidance meaning for preventing overuse of antibiotics.
Description
Technical field
The invention belongs to biological technical field, relate to the application of mushroom, particularly photosynthetic bacteria and bacillus subtilis prevent and treat the application in the agent of the red mouth germ of turbot in preparation.
Background technology
Turbot is to originate in the coastal a kind of famous and precious flatfish in Europe, the flat sub-circular of its health, and eyes are positioned at left side of body; Have a side to be blue or green brown, point-like melanin and a small amount of prickle are arranged, anophthalmia sidelight cunning is white in color; Dorsal fin and anal fin do not have hard thorn, and be very long, and head and tail fin are all less; Health middle part meat is thick, and visceral mass is little.The turbot natural distribution is littoral in Europe, east side, the Atlantic Ocean, is common in the North Sea and Baltic Sea.It has advantage such as adapt to that low water temperature life, fast growth, meat are good, breed and market potential are big, becomes one of good marine fish of European various countries exploitation in succession.Introduced China, and become the coastal important breed variety of northern China at present in 1992.But be accompanied by the expansion of breed scope and the increase of cultivation density, cause the disease problem of how precious fish to be on the rise, caused massive losses to aquaculture.Wherein " red mouth is sick ", main harm 50-150g fingerling, cardinal symptom shows as: the sick fish kiss end and the outside of belly upper jaw are hemorrhage; The part fish gill cover has the hemorrhage little pothole that festers, the oral cavity internal haemorrhage, and the enlargement of the sick flake eyeball of part is given prominence to, needled has a large amount of festers in the adventitia; The congested no food of enteron aisle; Di has a small amount of thick liquid, and liver is congested, the look dim diffusing shape of gruel that is, and the spleen enlargement is rubescent.In therapeutic process, the raiser is the economic pursuit benefit, and phenomenon such as medication happens occasionally carelessly, causes the residue of pesticide problem to be on the rise.How to carry out healthy aquaculture, aquatic products circle that just becomes problem demanding prompt solution.
Photosynthetic bacteria is one type of photoautotrophy or heterotrophic microorganism, can be that carbon source is carried out photosynthesis with the organic matter in the breeding water body, plays the effect that purifies water, suppresses the pathogenic microorganism growth and breeding and improve aquatic livestock immunity.
Withered grass bud pole bacterium is the Gram-positive bacteria that a kind of rod-short, no pod membrane can move, sprout in producing to embrace, for strictness aerobic.The distribution of withered grass bud pole bacterium is very extensive, and soil, lake, ocean, animals and plants body surface and habitat thereof all can be found.Be one of probio of using always, can secrete the growth that antibiotic suppresses pathogenic microorganism, avirulence, the application in aquatic products has great potentiality and advantage.
In conjunction with the mixed bacteria liquid that the advantage of photosynthetic bacteria and bacillus subtilis is processed, it is sick to reach the red mouth of effective control, and cure rate is up to 100%.Overcome side effect and the high frequency that the antibiotic abuse brings and changed expensive that water brings, a kind of safe, green, non-harmful aquaculture pattern is provided.
Summary of the invention
The objective of the invention is to overcome the weak point that existing abuse of antibiotics treatment fish disease causes various food-safety problems, provide a kind of photosynthetic bacteria and bacillus subtilis mixed bacteria liquid to prevent and treat the application in the agent of the red mouth germ of turbot in preparation.
The object of the invention is realized through following technical scheme:
Photosynthetic bacteria and bacillus subtilis mixed bacteria liquid are prevented and treated the application in the agent of the red mouth germ of turbot in preparation.
The ratio of photosynthetic bacteria and bacillus subtilis bacterial population is 1: 1 in said photosynthetic bacteria and the bacillus subtilis mixed bacteria liquid.
Said photosynthetic bacteria is red pseudomonas (Rhodopseudomonas sp.; CGMCC 1.2193), Rhodopseudomonas palustris (Rhodopseudomonas palustris; CGMCC 1.2352) or pod membrane red blood cell (Rhodobacter capsulatus, at least a in GIM1.168).
Said bacillus subtilis (Bacillus Subtilis Cohn, GIM1.136).
Said photosynthetic bacteria and the application of bacillus subtilis mixed bacteria liquid in turbot is formed are that photosynthetic bacteria and bacillus subtilis mixed bacteria liquid are splashed in water body, make that photosynthetic bacteria and bacillus subtilis mixed bacteria liquid final concentration are 10-10 in the water body
7Cfu/mL.
Said photosynthetic bacteria and bacillus subtilis mixed bacteria liquid are to use concentration to be 10-10 in feed in the application in turbot is formed
7The photosynthetic bacteria of cfu/mL and bacillus subtilis mixed bacteria liquid soak 15～45min.The present invention utilizes photosynthetic bacteria and bacillus subtilis to suppress the pathogenic microorganism growth and breeding, thereby realizes effectively preventing and treating the sick purpose of the red mouth of turbot.
The present invention has following advantage and effect with respect to prior art:
1, the present invention can effectively prevent and treat the red mouth disease of turbot.
2, microbial inoculum according to the invention safety in the application of disease control of aquatic animal is good.
Utilize photosynthetic bacteria and bacillus subtilis to the antagonism of pathogen, utilize organic substance in the culture environment of aquatic products, produce some and help the active factors of aquatic animal to grow and reach the method for preventing and treating disease belonging to biological method, thereby overcome the side effect that drug abuse brings
3, microbial inoculum of the present invention effectively, environmental protection, do not produce the pathogen resistance, application prospect is good.
Embodiment
Below in conjunction with embodiment the present invention is described in further detail, but embodiment of the present invention is not limited thereto.
The preparation of embodiment 1 bactericide
(1) preparation of photosynthetic bacteria liquid
Concentration is 10
9The red pseudomonas of cfu/mL (Rhodopseudomonas sp.; CGMCC 1.2193; Purchase in Chinese common micro-organisms culture presevation administrative center) be inoculated into by 0.1% of culture volume and optimize in the RCVBN liquid nutrient medium; In 30 ℃, intensity of illumination is that the illumination anaerobism is cultivated in the RCVBN medium that the bacterium liquid that obtains behind the 48h goes into to optimize by 10% inoculation of culture volume under the 3000lx; The illumination anaerobic fermentation is cultivated 48h, and the centrifugal 15min of 6000rpm obtains thalline then, and can to obtain concentration be 10 to the aseptic phosphate buffer of adding in the thalline
12The photosynthetic bacteria liquid of cfu/mL.
(2) preparation of bacillus subtilis bacterium liquid
Concentration is 10
9The bacillus subtilis of cfu/mL (Bacillus Subtilis Cohn; GIM1.136; Purchase in the Guangdong Microbes Inst) be inoculated in the nutrient broth medium by 0.1% of culture volume; In 37 ℃, 250rpm shaking table activation culture 18h, 10% inoculation that the bacterium liquid that obtains is mentioned by medium is gone in the nutrition broth bouillon, places 37 ℃, 250rpm shaking table cultivation 24h; The centrifugal 15min of 6000rpm obtains thalline then, and can to obtain concentration be 10 to the aseptic phosphate buffer of adding in the thalline
12The photosynthetic bacteria liquid of cfu/mL.
(3) the photosynthetic bacteria concentrate is mixed by bacterial population with the bacillus subtilis concentrate at 1: 1.
The application 1 of embodiment 2 bactericide
Each turbot is cultured pond turbot 5000 tails (turbot derives from the fish farm, Shandong Province), turbot counterpoise 25 ± 0.5g, and water temperature is 25 ℃.
Average mark A, B, C, four experimental group of D, B, C, each experimental group of D have 4 groups, are respectively B1, C1, D1; B2, C2, D2; B3, C3, D3; B4, C4, D4; Every group of 2 repetitions.The processing of five groups of experimental group is following:
A group: the breeding way of plant produced (control group).
B group: the mixed bacteria liquid of in water, splash photosynthetic bacteria and bacillus subtilis makes that photosynthetic bacteria and bacillus subtilis mixed bacteria liquid concentration reach 10,10 respectively in the water body
3, 10
5, 10
7Cfu/mL, the feeding normal diet changed Quan Chishui in per 40 days and adds bacterium after once.
The C group: in water, throwing in through concentration is 10,10
3, 10
5, 10
7Feed after the photosynthetic bacteria of cfu/mL and the mixed bacteria liquid of bacillus subtilis soak changed Quan Chishui once in per 40 days.
D group: in water, add the mixed bacteria liquid of photosynthetic bacteria and bacillus subtilis, make that photosynthetic bacteria and bacillus subtilis mixed bacteria liquid concentration reach 10,10 respectively in the water body
3, 10
5, 10
7Cfu/mL, throwing in through concentration simultaneously respectively is 10,10
3, 10
5, 10
7Feed after the photosynthetic bacteria of cfu/mL and bacillus subtilis mixed bacteria liquid soak changed Quan Chishui in per 40 days and adds bacterium after once.
The bacterial concentration that B1, C1, D1 throw in is 10cfu/mL
The bacterial concentration that B2, C2, D2 throw in is 10
3Cfu/mL
The bacterial concentration that B3, C3, D3 throw in is 10
5Cfu/mL
The bacterial concentration that B4, C4, D4 throw in is 10
7Cfu/mL
Feeds utilized People's Republic of China's agricultural industry criteria " the manufactured feed safety limit is used in the pollution-free food fishing " regulation (NY5072-2002) that meets.
Off-test is measured turbot and is suffered from the sick number of red mouth.
The sick illness rate of the red mouth of table one
Can find out that by last table through adding photosynthetic bacteria and bacillus subtilis bacterium liquid, it is sick to prevent turbot to change red mouth, can be effective as long as least concentration reaches 10cfu/mL.Thereby the breed new method of a kind of green, safety is provided for the breed of turbot.
The application 2 of embodiment 3 bactericide
Each turbot is cultured the pond and is suffered from the sick turbot of red mouth (the sick fish kiss end and the outside of belly upper jaw are hemorrhage) 30 tails, and water temperature is 25 ℃, 8 days treatment times.
Average mark A, B, C, D, five experimental group of E, C, D, each experimental group of E have 4 groups, are respectively C1, D1, E1; C2, D2, E2; C3, D3, E3; C4, D4, E4; C1, E1; C2, E2; C3, E3; C4, the every group of E4 are further divided into 2 groups, i.e. C11, C12......E41, E42, every group of 2 repetitions.The processing of five groups of experimental group is following:
A group: the breeding way of plant produced (control group).
The B group: the antibiotic of in water, splashing (PVP-I, 1mg/L-2mg/L), the feeding normal diet.
C group: the mixed bacteria liquid of in water, splash photosynthetic bacteria and bacillus subtilis makes that photosynthetic bacteria and bacillus subtilis mixed bacteria liquid concentration reach 10,10 respectively in the water body
3, 10
5, 10
7Cfu/mL, the feeding normal diet.
The D group: in water, throwing in through concentration is 10,10
3, 10
5, 10
7Feed after the photosynthetic bacteria of cfu/mL and the mixed bacteria liquid of bacillus subtilis soak.
E group: in water, add the mixed bacteria liquid of photosynthetic bacteria and bacillus subtilis, make cell concentration reach 10,10 respectively
3, 10
5, 10
7Cfu/mL, throwing in through concentration simultaneously respectively is 10,10
3, 10
5, 10
7Feed after the photosynthetic bacteria of cfu/mL and bacillus subtilis mixed bacteria liquid soak.
The bacterial concentration that C1, D1, E1 throw in is 10cfu/mL
The bacterial concentration that C2, D2, E2 throw in is 10
3Cfu/mL
The bacterial concentration that C3, D3, E3 throw in is 10
5Cfu/mL
The bacterial concentration that C4, D4, E4 throw in is 10
7Cfu/mL
Feeds utilized People's Republic of China's agricultural industry criteria " the manufactured feed safety limit is used in the pollution-free food fishing " regulation (NY5072-2002) that meets.Used fish medicine meets the regulation of People's Republic of China's agricultural industry criteria " pollution-free food drugs used in aquiculture usage criteria " (NY 5071-2002).
The sick fish cure rate (%) of the red mouth of table two
Can find out by last table; C, D, E group are significantly good than A and B group effect; And no matter be the bacterium liquid or add bacterium liquid of in the pond, splashing in the feed the inside; Can reach the result of treatment of cure rate 100%, and splash simultaneously mixed bacteria liquid and the feeding mixed bacteria liquid feed better effects if of soaking.Photosynthetic bacteria and bacillus subtilis are through antagonism, suppress growth of pathogenic bacteria.The mixed bacteria liquid of photosynthetic bacteria and bacillus subtilis has very big value and application potential in the sick treatment of the red mouth of turbot.
Embodiment of the present invention is not restricted to the described embodiments; Other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; All should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (1)
1. photosynthetic bacteria and bacillus subtilis mixed bacteria liquid are prevented and treated the application in the agent of the red mouth germ of turbot in preparation, it is characterized in that: said photosynthetic bacteria is at least a among red pseudomonas CGMCC 1.2193 (Rhodopseudomonas sp.), Rhodopseudomonas palustris CGMCC 1.2352 (Rhodopseudomonas palustris) or the red bacterium GIM1.168 of pod membrane (Rhodobacter capsulatus); Said bacillus subtilis GIM1.136 (Racillus Subtilis Cohn); The ratio of photosynthetic bacteria and bacillus subtilis bacterial population is 1: 1 in said photosynthetic bacteria and the bacillus subtilis mixed bacteria liquid.
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| CN2010102684356A CN101919408B (en) | 2010-08-31 | 2010-08-31 | Application of photosynthetic bacterium and bacillus subtilis mixed bacterial liquid in preparing microbial inoculum for preventing and controlling red mouth disease of turbot |
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| CN103583880B (en) * | 2013-10-18 | 2015-04-08 | 上海永凡生物科技有限公司 | Microbial fermentation feed for litopenaeus vannamei |
| CN103667109B (en) * | 2013-10-23 | 2016-02-03 | 大连市水产技术推广总站 | A kind of Rhodobacter capsulatus and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101264980A (en) * | 2008-04-23 | 2008-09-17 | 中国水产科学研究院南海水产研究所 | Sea water culture environment multiple state position biological restoring method |
| CN101269886A (en) * | 2008-05-05 | 2008-09-24 | 中国水产科学研究院黄海水产研究所 | Water quality scavenging agent for high-density cultivation system for penaeus orientalis |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101264980A (en) * | 2008-04-23 | 2008-09-17 | 中国水产科学研究院南海水产研究所 | Sea water culture environment multiple state position biological restoring method |
| CN101269886A (en) * | 2008-05-05 | 2008-09-24 | 中国水产科学研究院黄海水产研究所 | Water quality scavenging agent for high-density cultivation system for penaeus orientalis |
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