Microecological preparation and method for applying microecological preparation to industrial aquaculture of penaeus vannamei boone

Abstract

The invention relates to the technical field of culture, in particular to a microecological preparation and a method for applying the microecological preparation to industrial culture of penaeus vannamei boone, and the method comprises the following steps: step S1, separation, identification and safety analysis of intestinal tract or healthy culture water strains of penaeus vannamei boone; and S2, influence of the strain on the growth performance of the penaeus vannamei. According to the method, strains are screened and separated from intestinal tracts and culture water bodies of the penaeus vannamei, the safety of the strains is analyzed, the influence of the screened strains on the growth performance of the penaeus vannamei is evaluated by applying different types and numbers of strains and planting modes in the culture process, and the microecological preparation is developed by optimizing a thallus formula and a production process; the method is applied to industrial aquaculture of the penaeus vannamei and realizes green and healthy aquaculture of the penaeus vannamei.

Description

Technical Field

[0001] The invention relates to the technical field of aquaculture, and in particular to a microecological preparation and an application thereof in a method for factory-based aquaculture of white shrimp. Background Art

[0002] Whiteleg shrimp, also known as Penaeus vannamei, has the advantages of fast growth, delicious meat and high nutritional value. It is recognized as a high-yield shrimp variety in the world and is also the main shrimp variety cultivated in my country. At present, high-yield whiteleg shrimp is mainly cultivated in factories, which realizes the supervision and control of shrimp growth throughout the process, and improves the breeding efficiency and output. However, during the breeding process, the content of nitrogen, phosphorus, nitrate and nitrite in organic waste such as uningested feed residues and excrement increases during the degradation process, and it is very easy to breed a large number of pathogenic microorganisms, which in turn affects the water quality and is not conducive to the healthy growth of shrimp. Therefore, while the shrimp farming industry is developing rapidly, problems such as environmental pollution, water quality deterioration and frequent diseases that restrict the development of the industry need to be solved urgently.

[0003] At present, the strains of commercially available microecological agents are derived from terrestrial species. However, the intestinal microbial flora structure of white shrimp is quite different from that of terrestrial species, and the existing microecological preparations have a poor colonization effect in shrimp. At the same time, the body temperature of white shrimp is lower and the growth environment is significantly different from that of terrestrial animals. The bacteria are not resistant to stress and the strains have poor adaptability. Some strains have weak resistance to adverse environmental conditions such as temperature, acid-base environment, and oxygen concentration.

[0004] The intestinal microbial flora structure of Penaeus vannamei is quite different from that of terrestrial species, resulting in poor application of commercially available microecological preparations in Penaeus vannamei farming.

[0005] The abuse of antibiotics and chemical drugs in the process of high-density farming of white shrimp has destroyed the farming environment and the ecological balance of the normal flora in the intestines of the farmed shrimp, leading to frequent shrimp diseases.

[0006] To sum up the above problems, we propose a microecological preparation and its application in the factory farming method of white shrimp. Summary of the invention

[0007] The purpose of the present invention is to provide a microecological preparation and its application in a method for factory-scale breeding of white shrimp to solve the problems raised in the above-mentioned background technology.

[0008] To achieve the above object, the present invention provides the following technical solutions:

[0009] A method for analyzing strains in water body of Penaeus vannamei culture, characterized by comprising:

[0010] Step S1, isolation, identification and safety analysis of strains from the intestinal tract of Penaeus vannamei or healthy aquaculture water;

[0011] Step S11, screening and isolating strains in the intestines of Penaeus vannamei or in aquaculture water, and performing morphological identification of the strains;

[0012] Step S12: using molecular biology techniques to identify the isolated strains and conduct phylogenetic tree analysis;

[0013] Step S13, screening the physical and chemical properties and safety indicators of the strains for comprehensive evaluation;

[0014] Step S2, the effect of the strain on the growth performance of Penaeus vannamei;

[0015] Step S21, the influence of strain type and quantity on growth indexes of Penaeus vannamei;

[0016] Step S22, the effect of strain colonization method on the microbial colony and pathogen abundance in the intestine of Penaeus vannamei;

[0017] Step S23, the effect of the strain on the utilization rate of the feed of Penaeus vannamei, and the changes of ammonia nitrogen and nitrite indicators in the water body are analyzed at the same time.

[0018] Preferably, the step S11 specifically includes:

[0019] The bacterial cells in the intestines of Penaeus vannamei or in the aquaculture water were screened by the spreading culture method, and the morphological structure of the colonies was observed using optical microscopy technology to preliminarily classify the strains.

[0020] Preferably, the step S12 specifically includes:

[0021] The strain sequence was determined using 16SrRNA technology, and the obtained strains were sequenced to preliminarily determine the homologous species. Mega5.0 was used to construct a phylogenetic evolutionary tree to further determine the species.

[0022] Preferably, the step S13 specifically includes:

[0023] Step S131, screening the ability of the isolated strain to inhibit the growth of indicator bacteria by the minimum inhibitory concentration method, and determining the ability of the strain to secrete amylase and protease by spectrophotometry;

[0024] Step S132, the ammonia nitrogen content is measured by a water quality analyzer, and the nitrite nitrogen concentration is measured by visible spectrophotometry;

[0025] Step S133: acute toxicity experiment in mice to evaluate the safety of different strains.

[0026] Preferably, the step S2 specifically includes:

[0027] Step S21, weigh the body weight and liver weight of the white shrimp with an electronic balance, calculate the weight gain rate and liver-to-body ratio; use a vernier caliper to measure the body length, cephalothorax length, second leg length and intestine length, and calculate the shrimp length and intestine-to-body ratio;

[0028] Step S22, analyzing the intestinal bacterial colony structure of Penaeus vannamei using high-throughput determination technology, and determining the type and abundance changes of pathogenic bacteria using 16SrRNA technology and PCR technology;

[0029] Step S23, measuring the weight and feed consumption of Penaeus vannamei, and calculating the feed utilization rate.

[0030] A method for analyzing strains in water for breeding Penaeus vannamei, further comprising:

[0031] Step S3, development of microecological preparations and their application in the cultivation of Penaeus vannamei;

[0032] Step S31, analyzing the effects of the probiotic preparations prepared by different strain types and proportions on the growth performance of Penaeus vannamei, and determining the optimal probiotic preparation formula;

[0033] Step S32, studying the effects of different carriers and storage methods on the activity of the probiotic preparation, and determining the production process of the probiotic preparation;

[0034] Step S33: applying the microecological preparation to the factory-based farming of Penaeus vannamei according to the optimal microecological preparation formula.

[0035] Preferably, the screening of the optimal microecological preparation formula in step S31 specifically includes:

[0036] The compatibility of strain types and quantities, weight gain rate, body length, intestine-to-body ratio, feed utilization rate, and ammonia nitrogen and nitrite nitrogen indicators of white shrimp were analyzed, and the effects of different microecological preparation formulas were comprehensively evaluated.

[0037] Preferably, the production process of the microecological preparation in step S32 specifically includes:

[0038] Using different breeding feeds or embedding agents as strain carriers and storage conditions as factors, the number of bacteria was calculated using the spreading culture method, and the enzyme activity of the strain was determined by the spectrophotometer.

[0039] A microecological preparation is applied to the factory farming of Penaeus vannamei.

[0040] A microecological preparation is applied to a method for industrialized breeding of white shrimp, specifically comprising:

[0041] The weight gain rate, body length, intestine-to-body ratio, feed utilization rate, and ammonia nitrogen and nitrite nitrogen indicators of whiteleg shrimp were determined during the application of microecology, and the optimal microecological preparation was applied in the factory farming of whiteleg shrimp.

[0042] Compared with the prior art, the present invention has the following beneficial effects:

[0043] The invention uses the intestinal tract of Penaeus vannamei and healthy aquaculture water as raw materials to screen strains that can be well colonized and propagated in the intestinal tract of Penaeus vannamei.

[0044] The microecological preparation of the present invention has strong adaptability and can improve the growth performance of white shrimp and have a synergistic effect in regulating aquaculture water bodies.

[0045] The composite microecological preparation of the present invention has the advantages of low cost, no toxic side effects, no drug residue, no drug resistance, etc., and is an ecological and healthy breeding technology urgently needed for shrimp breeding production.

[0046] The invention screens and isolates strains from the intestinal tract of Penaeus vannamei and aquaculture water, analyzes the safety of the strains, applies strains of different types, quantities and colonization methods in the aquaculture process, evaluates the effects of the screened strains on the growth performance of Penaeus vannamei, optimizes the bacterial formula and production process to develop a microecological preparation, which is applied to the factory aquaculture of Penaeus vannamei, and realizes the green and healthy aquaculture of Penaeus vannamei. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 It is a schematic diagram of the structure of the present invention. DETAILED DESCRIPTION

[0048] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0049] Embodiment 1:

[0050] See also Figure 1 , a method for analyzing strains in water body of Penaeus vannamei culture, comprising:

[0051] Step S1, isolation, identification and safety analysis of strains from the intestinal tract of Penaeus vannamei or healthy aquaculture water;

[0052] Step S11, screening and isolating strains in the intestines of Penaeus vannamei or in aquaculture water, and performing morphological identification of the strains;

[0053] Step S12: using molecular biology techniques to identify the isolated strains and conduct phylogenetic tree analysis;

[0054] Step S13, screening the physical and chemical properties and safety indicators of the strains for comprehensive evaluation;

[0055] Step S2, the effect of the strain on the growth performance of Penaeus vannamei;

[0056] Step S21, the influence of strain type and quantity on growth indexes of Penaeus vannamei;

[0057] Step S22, the effect of strain colonization method on the microbial colony and pathogen abundance in the intestine of Penaeus vannamei;

[0058] Step S23, the effect of the strain on the utilization rate of the feed of Penaeus vannamei, and the changes of ammonia nitrogen and nitrite indicators in the water body are analyzed at the same time.

[0059] Embodiment 2:

[0060] like Figure 1 As shown, a microecological preparation is applied to the industrialized culture method of Penaeus vannamei, specifically including:

[0061] Step S1, isolation, identification and safety analysis of strains from the intestinal tract of Penaeus vannamei or healthy aquaculture water;

[0062] Step S11, screening and isolating strains in the intestines of Penaeus vannamei or in aquaculture water, and performing morphological identification of the strains;

[0063] The bacterial cells in the intestines of Penaeus vannamei or in the aquaculture water were screened by the spread culture method, and the morphological structure of the bacterial colonies was observed by optical microscopy to preliminarily classify the strains.

[0064] Step S12: using molecular biology techniques to identify the isolated strains and conduct phylogenetic tree analysis;

[0065] The strain sequence was determined using 16S rRNA technology, and the obtained strains were sequenced to preliminarily determine the homologous species. The phylogenetic evolutionary tree was constructed using Mega5.0 to further determine the species.

[0066] Step S13, screening the physical and chemical properties and safety indicators of the strains for comprehensive evaluation;

[0067] Step S131, screening the ability of the isolated strain to inhibit the growth of indicator bacteria by the minimum inhibitory concentration method, and determining the ability of the strain to secrete amylase and protease by spectrophotometry;

[0068] Step S132, the ammonia nitrogen content is measured by a water quality analyzer, and the nitrite nitrogen concentration is measured by visible spectrophotometry;

[0069] Step S133, acute toxicity experiment in mice to evaluate the safety of different strains;

[0070] Step S2, the effect of the strain on the growth performance of Penaeus vannamei;

[0071] Step S21, the influence of strain type and quantity on growth indexes of Penaeus vannamei;

[0072] The weight and liver weight of white shrimp were weighed with an electronic balance to calculate the weight gain rate and liver-to-body ratio; the body length, cephalothorax length, second leg length and intestine length were measured with a vernier caliper to calculate the shrimp length and intestine-to-body ratio;

[0073] Step S22, the effect of strain colonization method on the microbial colony and pathogen abundance in the intestine of Penaeus vannamei;

[0074] High-throughput measurement technology was used to analyze the intestinal flora structure of white shrimp, and 16SrRNA technology and PCR technology were used to determine the type and abundance changes of pathogens;

[0075] Step S23, the effect of the strain on the utilization rate of the feed of Penaeus vannamei, and the changes of ammonia nitrogen and nitrite indicators in the water body are analyzed;

[0076] Determine the weight and feed consumption of Penaeus vannamei and calculate feed utilization rate;

[0077] Step S3, development of microecological preparations and their application in the cultivation of Penaeus vannamei;

[0078] Step S31, analyzing the effects of the probiotic preparations prepared by different strain types and proportions on the growth performance of Penaeus vannamei, and determining the optimal probiotic preparation formula;

[0079] Analyze the compatibility of strain types and quantities, weight gain rate, body length, intestine-to-body ratio, feed utilization rate, and ammonia nitrogen and nitrite nitrogen indicators of white shrimp, and comprehensively evaluate the effects of different microecological preparations;

[0080] Step S32, studying the effects of different carriers and storage methods on the activity of the probiotic preparation, and determining the production process of the probiotic preparation;

[0081] Using different breeding feeds or embedding agents as strain carriers and storage conditions (storage temperature and time, feed moisture content) as factors, the number of bacteria was calculated using the spread culture method, and the enzyme activity of the strain was determined using the spectrophotometer method;

[0082] Step S33, applying the microecological preparation to the factory farming of Penaeus vannamei according to the optimal microecological preparation formula;

[0083] The weight gain rate, body length, intestine-to-body ratio, feed utilization rate, and ammonia nitrogen and nitrite nitrogen indicators of whiteleg shrimp were determined during the application of microecology, and the optimal microecological preparation was applied in the factory farming of whiteleg shrimp.

[0084] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for analyzing strains in water body of Penaeus vannamei culture, characterized in that: include: Step S1, isolation, identification and safety analysis of strains from the intestinal tract of Penaeus vannamei or healthy aquaculture water; Step S11, screening and isolating strains in the intestines of Penaeus vannamei or in aquaculture water, and performing morphological identification of the strains; Step S12: using molecular biology techniques to identify the isolated strains and conduct phylogenetic tree analysis; Step S13, screening the physical and chemical properties and safety indicators of the strains for comprehensive evaluation; Step S2, the effect of the strain on the growth performance of Penaeus vannamei; Step S21, the influence of strain type and quantity on growth indexes of Penaeus vannamei; Step S22, the effect of strain colonization method on the microbial colony and pathogen abundance in the intestine of Penaeus vannamei; Step S23, the effect of the strain on the utilization rate of the feed of Penaeus vannamei, and the changes of ammonia nitrogen and nitrite indicators in the water body are analyzed at the same time.

2. A method for analyzing strains in water body of Penaeus vannamei culture according to claim 1, characterized in that: The step S11 specifically includes: The bacterial cells in the intestines of Penaeus vannamei or in the aquaculture water were screened by the spreading culture method, and the morphological structure of the colonies was observed using optical microscopy technology to preliminarily classify the strains.

3. A method for analyzing strains in water body of Penaeus vannamei culture according to claim 1, characterized in that: The step S12 specifically includes: The strain sequence was determined using 16SrRNA technology, and the obtained strains were sequenced to preliminarily determine the homologous species. Mega5.0 was used to construct a phylogenetic evolutionary tree to further determine the species.

4. A method for analyzing strains in water body of Penaeus vannamei culture according to claim 1, characterized in that: The step S13 specifically includes: Step S131, screening the ability of the isolated strain to inhibit the growth of indicator bacteria by the minimum inhibitory concentration method, and determining the ability of the strain to secrete amylase and protease by spectrophotometry; Step S132, the ammonia nitrogen content is measured by a water quality analyzer, and the nitrite nitrogen concentration is measured by visible spectrophotometry; Step S133: acute toxicity experiment in mice to evaluate the safety of different strains.

5. The method for analyzing strains in water body of Penaeus vannamei culture according to claim 1, characterized in that: The step S2 specifically includes: Step S21, weigh the body weight and liver weight of the white shrimp with an electronic balance, calculate the weight gain rate and liver-to-body ratio; use a vernier caliper to measure the body length, cephalothorax length, second leg length and intestine length, and calculate the shrimp length and intestine-to-body ratio; Step S22, analyzing the intestinal bacterial colony structure of Penaeus vannamei using high-throughput determination technology, and determining the type and abundance changes of pathogenic bacteria using 16SrRNA technology and PCR technology; Step S23, measuring the weight and feed consumption of Penaeus vannamei, and calculating the feed utilization rate.

6. The method for analyzing strains in water body of Penaeus vannamei culture according to claim 1, characterized in that: Also includes: Step S3, development of microecological preparations and their application in the cultivation of Penaeus vannamei; Step S31, analyzing the effects of probiotics prepared by different strain types and proportions on the growth performance of white shrimp, and determining the optimal probiotic formula; Step S32, studying the effects of different carriers and storage methods on the activity of the probiotic preparation, and determining the production process of the probiotic preparation; Step S33: applying the microecological preparation to the factory-based farming of Penaeus vannamei according to the optimal microecological preparation formula.

7. A method for analyzing strains in water body of Penaeus vannamei culture according to claim 6, characterized in that: The screening of the optimal microecological preparation formula in step S31 specifically includes: The compatibility of strain types and quantities, weight gain rate, body length, intestine-to-body ratio, feed utilization rate, and ammonia nitrogen and nitrite nitrogen indicators of white shrimp were analyzed, and the effects of different microecological preparation formulas were comprehensively evaluated.

8. A method for analyzing strains in water body of Penaeus vannamei culture according to claim 7, characterized in that: The production process of the microecological preparation in step S32 specifically includes: Using different breeding feeds or embedding agents as strain carriers and storage conditions (storage temperature and time, feed moisture content) as factors, the number of bacteria was calculated using the spread culture method, and the enzyme activity of the strain was determined by the spectrophotometer.

9. A microecological preparation as claimed in claim 8 used in industrial farming of Penaeus vannamei.

10. A method for applying the probiotic preparation as claimed in claim 9 to a factory-based culture of Penaeus vannamei, characterized in that: Specific examples include: The weight gain rate, body length, intestine-to-body ratio, feed utilization rate, and ammonia nitrogen and nitrite nitrogen indicators of whiteleg shrimp were determined during the application of microecology, and the optimal microecological preparation was applied in the factory farming of whiteleg shrimp.

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