CN109157546A - A kind of compound micro-ecological preparation of the acute Hepatopancreatic necrosis syndrome of prevention and control Penaeus Vannmei and its application - Google Patents
A kind of compound micro-ecological preparation of the acute Hepatopancreatic necrosis syndrome of prevention and control Penaeus Vannmei and its application Download PDFInfo
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- CN109157546A CN109157546A CN201810844204.1A CN201810844204A CN109157546A CN 109157546 A CN109157546 A CN 109157546A CN 201810844204 A CN201810844204 A CN 201810844204A CN 109157546 A CN109157546 A CN 109157546A
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Abstract
The present invention relates to a kind of compound micro-ecological preparation of the acute Hepatopancreatic necrosis syndrome (AHPNS) of prevention and control Penaeus Vannmei and its applications, including Providence and enterobacteria.The compound micro-ecological preparation is added in feed and feeds.Compound micro-ecological preparation of the invention has effects that stronger inhibition vibrio parahemolyticus, has significant application effect, is one of the important channel for realizing Penaeus Vannmei green cultivation, has a good application prospect.
Description
Technical field
The invention belongs to aquatic products control and prevention of disease field, in particular to a kind of acute Hepatopancreatic necrosis of prevention and control Penaeus Vannmei
The compound micro-ecological preparation of syndrome and its application.
Background technique
The acute Hepatopancreatic necrosis syndrome (AHPNS) of Penaeus Vannmei is prominent to cause prawn with prawn Hepatopancreatic necrosis
Hair property steals a kind of dead disease for main feature, is to influence one of prawn culturing disease the most serious in recent years.Disease early stage
Prawn seedling stage is taken place mostly in, Deaths syndrome (Early mortality syndrome, EMS) is referred to as.It has sent out
Existing some vibrios for carrying specific Disease-causing gene are to lead to the pathogenetic cause of disease, wherein mainly vibrio parahaemolytious.Tran
Deng to from Vietnam AHPNS morbidity shrimp ponds in isolate 1 plant of Vibrio harveyi (Vibrio harveyi), impregnate infection after prawn it is dead
Rate is died up to 100%.Joshi etc. has isolated several plants of vibrio parahaemolytious from Thailand disease shrimp, and prawn is infected in intramuscular injection, wherein one group
Dead 100% in for 24 hours, bacterial strain detection is the AHPNS positive.Liu isolates 1 plant of Irving in the AHPNS morbidity shrimp ponds in Shanghai
Si Shi vibrios, it is believed that the bacterial strain is AHPNS pathogenic strain.Xuan etc. thinks that the 1 plant of Vibrio campbellii separated out of sick shrimp body can be led
Cause the generation of AHPNS.
PirvpAB gene coding binary toxin be pathogenic vibrio Major Virulence Factors, the binary toxin structure of generation with
Parasporal crystal structure caused by bacillus thuringiensis is similar, toxic effect also with bacillus thuringiensis Cry albumen virulence
Act on it is similar, all show as on Insertion Into Host Cell film formed wear die hole hole, intraor extracellular osmotic pressure changes, and makes target cell
Damage is even dead.
Existing part scientific research personnel starts the research and development for being conceived to probiotics, but it is acute to be specific to Penaeus Vannmei
The probiotics of Hepatopancreatic necrosis syndrome are delivered there is no any document at present.
Summary of the invention
Technical problem to be solved by the invention is to provide a kind of acute Hepatopancreatic necrosis syndromes of prevention and control Penaeus Vannmei
Compound micro-ecological preparation and its application, overcome in existing product that there are no bad specifically for the acute hepatopancrease of Penaeus Vannmei
The defect of the probiotics of dead syndrome.
The present invention provides a kind of compound micro-ecological preparation of the acute Hepatopancreatic necrosis syndrome of prevention and control Penaeus Vannmei, institutes
Stating complex ecological preparation includes Providence (Providencia sp.) and enterobacteria (Enterobacter sp.)；Its
In, Providence deposit number is CGMCC No.15415, and enterobacteria deposit number is CGMCC No.15416.
Providence (Providencia sp.) of the present invention, depositary institution's title: CGMCC China is micro-
Biological inoculum preservation administration committee common micro-organisms center, preservation address: in Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Institute of microbiology of the academy of sciences of state, deposit number are CGMCC NO.15415, and the deposit date is on March 6th, 2018.It is recommended that classification life
Name are as follows: Providence (Providencia sp.).
Enterobacteria (Enterobacter sp.) of the present invention, depositary institution's title: CGMCC China Microbiological bacterium
Kind preservation administration committee common micro-organisms center, preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Chinese science
Institute of microbiology of institute, deposit number are CGMCC NO.15416, and the deposit date is on March 6th, 2018.It is recommended that classification naming are as follows:
Enterobacteria (Enterobacter sp.).
The Providence and enterobacteria are cultivated respectively to concentration 1.0 × 108Mixing for standby use after CFU/mL.
The culture medium prescription of use are as follows: peptone 5g, yeast extract 1g, high ferric phosphate 0.01g, sodium acetate 3g, phosphoric acid hydrogen two
Potassium 1g, Chen Haishui 1000mL, pH value 7.6~7.8.
The configuration proportion of the Providence and enterobacteria after liquid fermentation is 1:0.5-2.
The ratio that the Providence and enterobacteria account for compound micro-ecological preparation is 2-3%.
The present invention also provides answering for the compound micro-ecological preparation of the acute Hepatopancreatic necrosis syndrome of prevention and control Penaeus Vannmei
With the compound micro-ecological preparation is added in feed and feeds.It can not be used in mixed way with antibiotic, dosage are as follows: per kilogram feed adds
Add 20~40ml of compound micro-ecological preparation, 3~5d of feeding cycle.
Present invention is mainly used for the acute Hepatopancreatic necrosis syndromes of prevention and control Penaeus Vannmei, and acute Hepatopancreatic necrosis is inhibited to cause
The growth of dead property vibrios, high mortality caused by reducing because of acute Hepatopancreatic necrosis syndrome.
Providence CGMCC No.15415 has the ability of synthesis or semi-synthetic beta-lactam antibiotics active material,
Lipopeptide antibiotic substance can be generated, compared with high inhibition Penaeus Vannmei pathogenic bacteria.
The adjustable intestinal flora balance of enterobacteria CGMCC No.15416 can produce class bacteriocin and lactic acid, inhibit cause of disease
Bacterium growth, improves Penaeus Vannmei immunity of organisms and premunition.
The two ratio according to the invention, which is applied in combination, enables to cause the acute Hepatopancreatic necrosis of Penaeus Vannmei to cause a disease
Property vibrio parahemolyticus rejection further enhance, in addition, the two collocation can inhibit 10 within the scope of Jiangsu Province thin
The pathogenic vibrio parahemolyticus of the acute Hepatopancreatic necrosis of the Penaeus Vannmei of bacterial strain type.And existing withered grass gemma currently on the market
The combination of bacillus and lactic acid bacteria can only inhibit conventional vibrio parahemolyticus to a certain extent, can not but inhibit multiple bacterial strains
The pathogenic parahemolyticas arc of the acute Hepatopancreatic necrosis of the Penaeus Vannmei of type.The combination of two plants of bacteriums of the invention, to Jiangsu Province's model
The pathogenic vibrio parahemolyticus of the acute Hepatopancreatic necrosis of Penaeus Vannmei of 10 bacterium plant types in enclosing has specific special
Property.
Beneficial effect
Compound micro-ecological preparation of the invention has effects that stronger inhibition vibrio parahemolyticus, has significant application
Effect is one of the important channel for realizing Penaeus Vannmei green cultivation, has a good application prospect.
Specific embodiment
Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention
Rather than it limits the scope of the invention.In addition, it should also be understood that, after reading the content taught by the present invention, those skilled in the art
Member can make various changes or modifications the present invention, and such equivalent forms equally fall within the application the appended claims and limited
Range.
Embodiment 1
Present embodiments provide a kind of compound micro-ecological preparation, including Providence (Providencia sp.) and
Enterobacteria (Enterobacter sp.)；Wherein, Providence deposit number is CGMCC No.15415, enterobacteria deposit number
For CGMCC No.15416.Two kinds of bacterium are formulated after liquid fermentation in 1:1 ratio respectively.Two kinds of bacterium cultivate respectively in advance to
Concentration 1.0 × 108Mixing for standby use after CFU/mL.Wherein, culture medium prescription are as follows: peptone 5g, yeast extract 1g, high ferric phosphate
0.01g, sodium acetate 3g, dipotassium hydrogen phosphate 1g, Chen Haishui 1000mL, pH value 7.6~7.8.
The application method of compound micro-ecological preparation is as follows:
Penaeus Vannmei pellet is added in Providence and enterobacteria culture mixed liquor, mixed proportion 1:
50, by the feedstuff feeding Penaeus Vannmei of mixing, and observe the food situation of bait table.The same day of every morning 8:00~9:00
It feeds for the first time, feeds daily once, feeding time 2 hours, feed 7d altogether.
The effect experiment of compound micro-ecological preparation is as follows:
One, inhibit acute Hepatopancreatic necrosis lethal vibrios test
9 shrimp seedling sources of In A Certain Locality, Jiangsu Province, the consistent prawn culturing temperature canopy of growth conditions are chosen in test, and infection outburst is acute
Liver Pancreatic Necrosis syndrome (about 0.7 mu/canopy) is randomly divided into 3 groups, and one group of control (is not added by the raising of raiser's normal mode
Any probiotics), it compares two groups and adds bacillus subtilis and lactic acid bacteria in feed, add general sieve in test group feed
Wei Dengsi bacterium and enterobacteria compound micro-ecological preparation, taken at the 0th, 1,3,5,7 day in pond fall ill shrimp sample (each pond takes at 3
The shrimp sample of different location takes the liver pancreas in shrimp sample, mixes), using molecular biology method, to acute in liver pancreatic tissues
Hepatopancreatic necrosis lethal vibrios carries out qualitative analysis, while observing prawn death condition.
Amount of vibrio changes in 1 water body of table
Note: different letters indicate significant difference (P<0.05), and same letter indicates that difference is not significant (P>0.05)
From table 1 it follows that one group of control is not added with any probiotics, acute Hepatopancreatic necrosis is comprehensive in shrimp body
Cause of disease always exists, the extremely significant increase of dead individuals quantity, until the 5th day all dead.It compares two groups and uses bacillus subtilis
Bacterium and lactic acid bacteria are as compound microecological system, and the comprehensive sick cause of disease of acute Hepatopancreatic necrosis always exists in shrimp body, dead individuals number
Extremely significant increase is measured, until the 5th day all dead, does not have significant difference with compareing one group.Test group feeds compound microecological system
After agent the 3rd day, the comprehensive sick Pathogen test of acute Hepatopancreatic necrosis is in weakly positive, is negative after the 5th day, dead individuals were from the 3rd day
After start conspicuousness reduction, have no dead individuals within the 7th day, before use compared with significant difference (P < 0.05).It can be seen that this reality
The using effect for applying example is significantly better than control group one and two.
Two, kinds of pathogenic vibrio plate Inhibition test
Test choose 3 plants of acute Hepatopancreatic necrosis syndrome lethal vibrios, with bite vibrios, bacillus, lactic acid bacteria and
Antagonistic experiment is arranged in this compound fermented liquid.Four groups are randomly divided into, the first control group is bacteriophage, and the second control group is gemma bar
Bacterium, third control group are lactic acid bacteria, and experimental group is this compound bacteria.3 pathogen strain bacterium fall ill inside the province from Jiangsu shrimp separation, purifying, warp
Molecular biology identification contains acute Hepatopancreatic necrosis syndrome virulence gene pirA and pirB, number HYS-AHPND-1,
HYS-AHPND-2, HYS-AHPND-3, three plants of bacterium are with bacteria concentration for 106CFU/ml is coated on LB plate, with four kinds of microorganisms
Fermentation liquid (biting vibrios, bacillus, lactic acid bacteria and this compound bacteria) is cooked drug sensitive test paper, is cultivated 48 hours at 28 DEG C, is measured antibacterial
The diameter of circle, experimental result are shown in Table 2.The result shows that compound formulation of the invention is to three plants of Jiangsu Penaeus Vannmei Acute Hepatic pancreas
The fungistatic effect of gland necrosis syndrome lethal vibrios it is extremely significant be higher than bite vibrios, bacillus, lactic acid bacteria (P < 0.01).
Bacteriostatic activity of 2 bacterial strain of table to the acute Hepatopancreatic necrosis syndrome lethal bacterial strain of Penaeus Vannmei
Note: different letters indicate significant difference (P<0.05), and same letter indicates that difference is not significant (P>0.05).
Claims (6)
1. a kind of compound micro-ecological preparation of the acute Hepatopancreatic necrosis syndrome of prevention and control Penaeus Vannmei, it is characterised in that: described
Complex ecological preparation includes Providence (Providencia sp.) and enterobacteria (Enterobacter sp.)；Wherein,
Providence deposit number is CGMCC No.15415, and enterobacteria deposit number is CGMCC No.15416.
2. a kind of compound microecological system of the acute Hepatopancreatic necrosis syndrome of prevention and control Penaeus Vannmei according to claim 1
Agent, it is characterised in that: the Providence and enterobacteria are cultivated respectively to concentration 1.0 × 108Mixing for standby use after CFU/mL.
3. a kind of compound microecological system of the acute Hepatopancreatic necrosis syndrome of prevention and control Penaeus Vannmei according to claim 2
Agent, it is characterised in that: the culture medium prescription of use are as follows: peptone 5g, yeast extract 1g, high ferric phosphate 0.01g, sodium acetate 3g, phosphorus
Sour hydrogen dipotassium 1g, Chen Haishui 1000mL, pH value 7.6~7.8.
4. a kind of compound microecological system of the acute Hepatopancreatic necrosis syndrome of prevention and control Penaeus Vannmei according to claim 1
Agent, it is characterised in that: the configuration proportion of the Providence and enterobacteria after liquid fermentation is 1:0.5-2.
5. a kind of compound microecological system of the acute Hepatopancreatic necrosis syndrome of prevention and control Penaeus Vannmei according to claim 1
Agent, it is characterised in that: the Providence and enterobacteria account for the ratio of compound micro-ecological preparation as 2-3%.
6. a kind of compound micro-ecological preparation of the acute Hepatopancreatic necrosis syndrome of prevention and control Penaeus Vannmei as described in claim 1
Application, it is characterised in that: the compound micro-ecological preparation is added in feed and feeds.
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| CN201810844204.1A CN109157546B (en) | 2018-07-27 | 2018-07-27 | Compound microecological preparation for preventing and controlling acute hepatopancreas necrosis syndrome of penaeus vannamei boone and application thereof |
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| CN201810844204.1A CN109157546B (en) | 2018-07-27 | 2018-07-27 | Compound microecological preparation for preventing and controlling acute hepatopancreas necrosis syndrome of penaeus vannamei boone and application thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112753629A (en) * | 2020-12-30 | 2021-05-07 | 亚太海洋生物科技(厦门)有限公司 | Method for removing acute hepatopancreas necrosis virus in early stage of culture of penaeus vannamei boone |
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| AU2003225946A1 (en) * | 2002-03-21 | 2003-10-08 | Board Of Regents, The University Of Texas System | Methods and compositions for vaccination against or involvingenterobacteriaceae bacteria |
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Patent Citations (4)
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|---|---|---|---|---|
| AU2003225946A1 (en) * | 2002-03-21 | 2003-10-08 | Board Of Regents, The University Of Texas System | Methods and compositions for vaccination against or involvingenterobacteriaceae bacteria |
| CN103340277A (en) * | 2013-06-07 | 2013-10-09 | 北京大北农科技集团股份有限公司 | Feeding microecological additive and compound feed containing same |
| CN103966122A (en) * | 2014-04-18 | 2014-08-06 | 中国科学院南海海洋研究所 | Providencia vermicola and application of providencia vermicola in preparing antibacterial drugs |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112753629A (en) * | 2020-12-30 | 2021-05-07 | 亚太海洋生物科技(厦门)有限公司 | Method for removing acute hepatopancreas necrosis virus in early stage of culture of penaeus vannamei boone |
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