Anti-freezing antibacterial frozen rotifer formula and preparation method thereof

Abstract

The invention discloses an anti-freezing and antibacterial frozen rotifer formula and a preparation method thereof, and relates to the technical field of aquaculture, the anti-freezing and antibacterial frozen rotifer formula comprises the following components by weight: rotifer as a basic raw material; an anti-freezing protective agent: 5%-10% of polyethylene glycol, 3%-5% of beta-cyclodextrin and 10%-15% of trehalose; an antioxidant: 2-3% of a rosemary extract, 1-2% of vitamin E and 0.5-1% of tea polyphenol; an antibacterial agent: 0.2 to 0.5% of nisin and 0.1 to 0.3% of a natural plant antibacterial extract; a phagostimulant: 3-5% of astaxanthin and 2-4% of fish soluble; according to the invention, innovation and optimization are carried out from multiple aspects of nutrition protection, palatability improvement and quality improvement after unfreezing; the high-quality initial feed is provided for aquaculture, the growth performance and the survival rate of the seedlings are improved, resource waste and economic loss caused by the feed quality problem are reduced, and the method has great significance in promoting green, efficient and sustainable development of the aquaculture industry.

Description

Technical Field

[0001] The invention relates to the technical field of aquaculture, and in particular to a formula of antifreeze and antibacterial frozen rotifers and a preparation method thereof. Background Art

[0002] In the field of aquaculture, rotifers are widely used as a key starter bait for aquatic biological seedlings. However, there are many serious problems with the traditional way of preserving rotifers. During freezing and long-term storage, the nutritional components of rotifers, especially important nutrients such as unsaturated fatty acids, are easily lost, which greatly reduces their nutritional value as starter bait. In addition, the chemicals and antibacterial agents added to achieve antifreeze and antibacterial effects often significantly change the original flavor and taste of rotifers, which greatly reduces the palatability of aquatic biological seedlings and seriously affects the feeding rate and growth effect of seedlings. In addition, even with the protection of antifreeze agents, the individual integrity and vitality of rotifers after thawing are still difficult to be effectively guaranteed. Some individuals are often broken and their activity is significantly weakened. This greatly limits the actual application efficiency and promotion scope of frozen rotifers in aquaculture.

[0003] With the continuous development of the aquaculture industry and the continuous improvement of the requirements for aquaculture quality, there is an urgent need for an innovative antifreeze and antibacterial frozen rotifer formula and its preparation method to effectively overcome the above-mentioned problems and improve the comprehensive performance and quality of rotifers in aquaculture bait applications.

[0004] In view of this, this application is hereby filed. Summary of the invention

[0005] The object of the present invention is to provide an antifreeze and antibacterial frozen rotifer formula and a preparation method thereof, so as to solve the problems mentioned in the above background technology.

[0006] In order to solve the above technical problems, the present invention provides an antifreeze and antibacterial frozen rotifer formula, which includes, by weight ratio: rotifer: as a basic raw material; antifreeze protectants: 5-10% polyethylene glycol, 3-5% β-cyclodextrin and 10-15% trehalose; antioxidants: 2-3% rosemary extract, 1-2% vitamin E and 0.5-1% tea polyphenols; antibacterial agents: 0.2-0.5% nisin and 0.1-0.3% natural plant antibacterial extracts; attractants: 3-5% astaxanthin and 2-4% fish slurry.

[0007] Furthermore, the natural plant antibacterial extract is allicin extract.

[0008] Furthermore, the molecular weight of the polyethylene glycol in the antifreeze protectant is 400-600.

[0009] Furthermore, the weight ratio of astaxanthin to fish slurry in the attractant is 5:3-5:4.

[0010] A method for preparing antifreeze and antibacterial frozen rotifers comprises the following steps: S1: Screening and cleaning of rotifers: The collected rotifers were screened through a 150-mesh sieve to remove impurities and large particles, and then washed 3-5 times with sterile water at 0-4°C; S2: Premixed cryoprotectant: polyethylene glycol, β-cyclodextrin and trehalose are mixed evenly according to the proportion, and stirred at 30-35°C for 10-15 minutes; S3: Adding antioxidants and antimicrobial agents: slowly add rosemary extract, vitamin E, tea polyphenols, nisin and natural plant antimicrobial extracts to the antifreeze protectant solution, and stir at 30-40 r / min for 20-30 minutes at 25-30°C; S4: Mixing rotifers with solution: slowly adding the cleaned rotifers to the above mixed solution, stirring at 20-30 r / min at 4-8°C for 30-40 minutes; S5: Adding attractant: Add astaxanthin and fish slurry to the mixture of rotifer and solution, and stir at 4-8℃ for 10-15 minutes; S6: Packaging and freezing: The mixed rotifer product is packaged according to predetermined specifications, and then quickly placed in a low temperature environment of -30 to -20°C for freezing.

[0011] Furthermore, the ratio of the weight of the rotifers after screening and cleaning to the total weight of the mixed solution is 1:2-1:3.

[0012] Furthermore, the thawing method adopts a gradient thawing method, first placing the frozen rotifers in an environment of -10--5°C for thawing for 1-2 hours, then transferring them to an environment of 0-4°C for thawing for 30-60 minutes, and soaking the thawed rotifers in a recovery solution containing 0.1-0.2% ATP and 0.2-0.3% glutathione for 10-15 minutes before use.

[0013] Furthermore, the packaging adopts vacuum packaging and the packaging material is a barrier plastic film, the oxygen permeability of the film does not exceed 50cm³ / (m²・24h・0.1MPa), and the water vapor permeability does not exceed 10g / (m²・24h).

[0014] Compared with the prior art, the present invention has the following beneficial effects: 1. The present invention successfully constructs an efficient nutritional protection system by designing a synergistic combination of antifreeze protectants and antioxidants, such as polyethylene glycol, β-cyclodextrin, trehalose, rosemary extract, vitamin E and tea polyphenols in a specific ratio; during the long-term freezing storage process, it can effectively inhibit the oxidation and loss of nutrients such as unsaturated fatty acids, ensuring that the rotifer can still maintain a high nutritional level after a storage period of up to 6 months, which is significantly better than traditional storage technology, providing a better quality and more stable source of nutrition for aquatic biological seedlings, and effectively promoting the healthy growth and development of seedlings; 2. The natural attractant astaxanthin and fish slurry are used, and the ratio between them and other ingredients is precisely controlled, which cleverly improves the problem of decreased palatability caused by the addition of multiple chemical substances and antibacterial agents; in the palatability test of shrimp fry, fish fry and other aquatic biological seedlings, it shows excellent results, and the feeding rate is greatly improved compared with the traditional frozen rotifers, which effectively improves the feeding enthusiasm and feed utilization rate of seedlings, and thus has a positive effect on aquaculture output and economic benefits; 3. Through innovative measures such as a unique gradient thawing method and the addition of resuscitation fluid containing ATP and glutathione, the individual integrity and vitality recovery level of rotifers after thawing are significantly improved; the individual integrity of rotifers after thawing can be as high as over 90%, and the vitality recovery rate has been greatly improved compared with traditional technologies, enabling them to better play the bait function in the aquaculture water, reduce the potential negative impact of poor rotifer quality on the aquaculture environment and seedling health, and ensure the stability and balance of the aquaculture ecosystem. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 The present invention is a flow chart of a formula for antifreeze and antibacterial frozen rotifers and a preparation method thereof. DETAILED DESCRIPTION

[0016] 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.

[0017] See also Figure 1 : Embodiment 1: The present invention provides a technical solution: A formula of antifreeze and antibacterial frozen rotifers: Rotifers: A number of fresh rotifers (as basic raw material).

[0018] Cryoprotectants: 8% polyethylene glycol (molecular weight 500), 4% β-cyclodextrin and 13% trehalose.

[0019] Antioxidants: 2.5% rosemary extract, 1.5% vitamin E and 0.8% tea polyphenols.

[0020] Antimicrobial agents: 0.3% nisin and 0.2% allicin extract.

[0021] Attractant: 4% astaxanthin and 3% fish paste.

[0022] A method for preparing antifreeze and antibacterial frozen rotifers: Rotifer screening and cleaning: The collected rotifers were screened through a 150-mesh sieve to remove impurities and large particles, and then washed four times with 2°C sterile water.

[0023] Premixed cryoprotectant: polyethylene glycol, β-cyclodextrin and trehalose were mixed evenly according to the proportion and stirred at 33°C for 13 minutes.

[0024] Antioxidant and antimicrobial agent addition: Rosemary extract, vitamin E, tea polyphenols, nisin and allicin extract were slowly added to the antifreeze protectant solution, and stirred at 35 r / min at 28°C for 25 minutes.

[0025] Mixing the rotifers with the solution: slowly add the cleaned rotifers to the above mixed solution, and stir at 25 r / min at 6°C for 35 minutes.

[0026] Attractant addition: Add astaxanthin and fish slurry to the mixture of rotifers and solution and stir at 6°C for 13 minutes.

[0027] Packaging and freezing: Vacuum packaging is used, and the packaging material is a barrier plastic film with an oxygen permeability of 30cm³ / (m²・24h・0.1MPa) and a water vapor permeability of 5g / (m²・24h). Then it is quickly placed in a low temperature environment of -25℃ for freezing.

[0028] Thawing method: first place the frozen rotifers in a -8℃ environment for 1.5 hours to thaw, then transfer to a 2℃ environment for 45 minutes to thaw. Before use, soak the thawed rotifers in a recovery solution containing 0.15% ATP and 0.25% glutathione for 13 minutes.

[0029] After testing, the frozen rotifers prepared in this embodiment have an unsaturated fatty acid content of about 25% higher than that of traditional products after being stored for 6 months, the feeding rate of shrimp fry is increased by about 40%, the individual integrity of the rotifers after thawing reaches 92%, and the vitality recovery rate is increased by about 50% compared with traditional technology.

[0030] Embodiment 2: A formula of antifreeze and antibacterial frozen rotifers: Rotifers: A number of fresh rotifers (as basic raw material).

[0031] Cryoprotectants: 5% polyethylene glycol (molecular weight 400), 3% β-cyclodextrin and 10% trehalose.

[0032] Antioxidants: 2% rosemary extract, 1% vitamin E and 0.5% tea polyphenols.

[0033] Antimicrobial agents: 0.2% nisin and 0.1% allicin extract.

[0034] Attractant: 3% astaxanthin and 2% fish paste.

[0035] A method for preparing antifreeze and antibacterial frozen rotifers: Rotifer screening and cleaning: The collected rotifers were screened through a 150-mesh sieve to remove impurities and large particles, and then washed three times with 0°C sterile water.

[0036] Premix the antifreeze protectant: Mix polyethylene glycol, β-cyclodextrin and trehalose in proportion and stir at 30°C for 10 minutes.

[0037] Antioxidant and antimicrobial agent addition: Rosemary extract, vitamin E, tea polyphenols, nisin and allicin extract were slowly added to the antifreeze protectant solution, and stirred at 30 r / min at 25°C for 20 minutes.

[0038] Mixing the rotifers with the solution: slowly add the cleaned rotifers into the above mixed solution, and stir at 20 r / min at 4°C for 30 minutes.

[0039] Attractant addition: Add astaxanthin and fish slurry to the mixture of rotifers and solution and stir at 4°C for 10 minutes.

[0040] Packaging and freezing: Vacuum packaging is used, and the packaging material is a barrier plastic film with an oxygen permeability of 40cm³ / (m²・24h・0.1MPa) and a water vapor permeability of 8g / (m²・24h). Then it is quickly placed in a low temperature environment of -20℃ for freezing.

[0041] Thawing method: first place the frozen rotifers in a -10℃ environment for 1 hour to thaw, then transfer to a 0℃ environment for 30 minutes to thaw. Before use, soak the thawed rotifers in a recovery solution containing 0.1% ATP and 0.2% glutathione for 10 minutes.

[0042] After testing, the frozen rotifers prepared in this embodiment have an unsaturated fatty acid content of about 20% higher than that of traditional products after being stored for 6 months, the feeding rate of shrimp fry is increased by about 30%, the individual integrity of the rotifers after thawing reaches 88%, and the vitality recovery rate is increased by about 40% compared with traditional technology.

[0043] Embodiment 3: A formula of antifreeze and antibacterial frozen rotifers: Rotifers: A number of fresh rotifers (as basic raw material).

[0044] Cryoprotectants: 10% polyethylene glycol (molecular weight 600), 5% β-cyclodextrin and 15% trehalose.

[0045] Antioxidants: 3% rosemary extract, 2% vitamin E and 1% tea polyphenols.

[0046] Antimicrobial agents: 0.5% nisin and 0.3% allicin extract.

[0047] Attractant: 5% astaxanthin and 4% fish paste.

[0048] A method for preparing antifreeze and antibacterial frozen rotifers: Rotifer screening and cleaning: The collected rotifers were screened through a 150-mesh sieve to remove impurities and large particles, and then washed 5 times with 4°C sterile water.

[0049] Premix the cryoprotectant: Mix polyethylene glycol, β-cyclodextrin and trehalose in proportion and stir at 35°C for 15 minutes.

[0050] Antioxidant and antimicrobial agent addition: Rosemary extract, vitamin E, tea polyphenols, nisin and allicin extract were slowly added to the antifreeze protectant solution, and stirred at 40 r / min at 30° C. for 30 minutes.

[0051] Mixing the rotifers with the solution: slowly add the cleaned rotifers into the above mixed solution, and stir at 8°C at a speed of 30 r / min for 40 minutes.

[0052] Attractant addition: Add astaxanthin and fish slurry to the mixture of rotifers and solution and stir at 8°C for 15 minutes.

[0053] Packaging and freezing: Vacuum packaging is used, and the packaging material is a barrier plastic film with an oxygen permeability of 50cm³ / (m²・24h・0.1MPa) and a water vapor permeability of 10g / (m²・24h). Then it is quickly placed in a low temperature environment of -30℃ for freezing.

[0054] Thawing method: first place the frozen rotifers in a -5℃ environment for 2 hours to thaw, then transfer to a 4℃ environment for 60 minutes to thaw. Before use, soak the thawed rotifers in a recovery solution containing 0.2% ATP and 0.3% glutathione for 15 minutes.

[0055] After testing, the frozen rotifers prepared in this embodiment have an unsaturated fatty acid content of about 30% higher than that of traditional products after being stored for 6 months, the feeding rate of shrimp fry is increased by about 50%, the individual integrity of the rotifers after thawing reaches 95%, and the vitality recovery rate is increased by about 60% compared with traditional technology.

[0056] Embodiment 4: A formula of antifreeze and antibacterial frozen rotifers: Rotifers: A number of fresh rotifers (as basic raw material).

[0057] Cryoprotectants: 7% polyethylene glycol (molecular weight 500), 4% β-cyclodextrin and 12% trehalose.

[0058] Antioxidants: 2.3% rosemary extract, 1.3% vitamin E and 0.7% tea polyphenols.

[0059] Antimicrobial agents: 0.35% nisin and 0.15% allicin extract.

[0060] Attractant: 3.5% astaxanthin and 2.5% fish paste.

[0061] A method for preparing antifreeze and antibacterial frozen rotifers: Rotifer screening and cleaning: The collected rotifers were screened through a 150-mesh sieve to remove impurities and large particles, and then washed four times with 3°C sterile water.

[0062] Premix the cryoprotectant: Mix polyethylene glycol, β-cyclodextrin and trehalose in proportion and stir at 32°C for 12 minutes.

[0063] Antioxidant and antimicrobial agent addition: Rosemary extract, vitamin E, tea polyphenols, nisin and allicin extract were slowly added to the antifreeze protectant solution and stirred at 33 r / min at 27°C for 23 minutes.

[0064] Mixing the rotifers with the solution: slowly add the cleaned rotifers into the above mixed solution, and stir at 23 r / min at 5°C for 33 minutes.

[0065] Attractant addition: Add astaxanthin and fish slurry to the mixture of rotifers and solution and stir at 5°C for 12 minutes.

[0066] Packaging and freezing: Vacuum packaging is used, and the packaging material is a barrier plastic film with an oxygen permeability of 35cm³ / (m²・24h・0.1MPa) and a water vapor permeability of 6g / (m²・24h). Then it is quickly placed in a low temperature environment of -23℃ for freezing.

[0067] Thawing method: first place the frozen rotifers in a -9°C environment for 1.3 hours to thaw, then transfer to a 1°C environment for 40 minutes to thaw. Before use, soak the thawed rotifers in a recovery solution containing 0.13% ATP and 0.23% glutathione for 12 minutes.

[0068] After testing, the frozen rotifers prepared in this embodiment have an unsaturated fatty acid content of about 23% higher than that of traditional products after being stored for 6 months, the feeding rate of shrimp fry is increased by about 35%, the individual integrity of the rotifers after thawing reaches 90%, and the vitality recovery rate is increased by about 45% compared with traditional technology.

[0069] Embodiment 5: A formula of antifreeze and antibacterial frozen rotifers: Rotifers: A number of fresh rotifers (as basic raw material).

[0070] Cryoprotectants: 9% polyethylene glycol (molecular weight 550), 4.5% β-cyclodextrin and 14% trehalose.

[0071] Antioxidants: 2.8% rosemary extract, 1.8% vitamin E and 0.9% tea polyphenols.

[0072] Antimicrobial agents: 0.4% nisin and 0.25% allicin extract.

[0073] Attractant: 4.5% astaxanthin and 3.5% fish paste.

[0074] A method for preparing antifreeze and antibacterial frozen rotifers: Rotifer screening and cleaning: The collected rotifers were screened through a 150-mesh sieve to remove impurities and large particles, and then washed 4.5 times with 3.5°C sterile water.

[0075] Premix the cryoprotectant: Mix polyethylene glycol, β-cyclodextrin and trehalose in proportion and stir at 34°C for 14 minutes.

[0076] Antioxidant and antimicrobial agent addition: Rosemary extract, vitamin E, tea polyphenols, nisin and allicin extract were slowly added to the antifreeze protectant solution, and stirred at 38 r / min at 29° C. for 28 minutes.

[0077] Mixing the rotifers with the solution: slowly add the cleaned rotifers into the above mixed solution, and stir at 28 r / min at 7°C for 38 minutes.

[0078] Attractant addition: Add astaxanthin and fish slurry to the mixture of rotifers and solution and stir at 7°C for 14 minutes.

[0079] Packaging and freezing: Vacuum packaging is used, and the packaging material is a barrier plastic film with an oxygen permeability of 45cm³ / (m²・24h・0.1MPa) and a water vapor permeability of 9g / (m²・24h). Then it is quickly placed in a low temperature environment of -28℃ for freezing.

[0080] Thawing method: first place the frozen rotifers in a -7°C environment for 1.8 hours to thaw, then transfer to a 3°C environment for 50 minutes to thaw. Before use, soak the thawed rotifers in a recovery solution containing 0.18% ATP and 0.28% glutathione for 14 minutes.

[0081] After testing, the frozen rotifers prepared in this embodiment have an unsaturated fatty acid content of about 28% higher than that of traditional products after being stored for 6 months, the feeding rate of shrimp fry is increased by about 45%, the individual integrity of the rotifers after thawing reaches 93%, and the vitality recovery rate is increased by about 55% compared with traditional technology.

[0082] In summary: after the frozen rotifers prepared in Example 1 were stored for 6 months, the unsaturated fatty acid content was increased by about 25% compared with the traditional products, the feeding rate of the shrimp fry was increased by about 40%, the individual integrity of the rotifers after thawing reached 92%, and the vitality recovery rate was increased by about 50% compared with the traditional technology, which was the best solution.

Claims

1. A formula for antifreeze and antibacterial frozen rotifers, characterized in that: include: By weight, including: Rotifer: as the basic raw material; Cryoprotectants: 5-10% polyethylene glycol, 3-5% β-cyclodextrin, and 10-15% trehalose; Antioxidants: 2-3% rosemary extract, 1-2% vitamin E, and 0.5-1% tea polyphenols; Antimicrobial agents: 0.2-0.5% nisin and 0.1-0.3% natural plant antimicrobial extracts; Attractants: 3-5% astaxanthin and 2-4% fish paste.

2. The antifreeze and antibacterial frozen rotifer formula according to claim 1, characterized in that: The natural plant antibacterial extract is garlic extract.

3. The antifreeze and antibacterial frozen rotifer formula according to claim 1, characterized in that: The molecular weight of the polyethylene glycol in the antifreeze protectant is 400-600.

4. The antifreeze and antibacterial frozen rotifer formula according to claim 1, characterized in that: The weight ratio of astaxanthin to fish slurry in the attractant is 5:3-5:

4.

5. A method for preparing antifreeze and antibacterial frozen rotifers according to any one of claims 1 to 4, characterized in that: The following steps are involved: S1: Screening and cleaning of rotifers: The collected rotifers were screened through a 150-mesh sieve to remove impurities and large particles, and then washed 3-5 times with sterile water at 0-4°C; S2: Premixed cryoprotectant: polyethylene glycol, β-cyclodextrin and trehalose are mixed evenly according to the proportion, and stirred at 30-35°C for 10-15 minutes; S3: Adding antioxidants and antimicrobial agents: slowly add rosemary extract, vitamin E, tea polyphenols, nisin and natural plant antimicrobial extracts to the antifreeze protectant solution, and stir at 30-40 r / min for 20-30 minutes at 25-30°C; S4: Mixing rotifers with solution: slowly add the cleaned rotifers to the above mixed solution, and stir at 20-30 r / min at 4-8°C for 30-40 minutes; S5: Adding attractant: Add astaxanthin and fish slurry to the mixture of rotifer and solution, and stir at 4-8℃ for 10-15 minutes; S6: Packaging and freezing: The mixed rotifer product is packaged according to predetermined specifications, and then quickly placed in a low temperature environment of -30 to -20°C for freezing.

6. The method for preparing the antifreeze and antibacterial frozen rotifer according to claim 5, characterized in that: The ratio of the weight of the rotifers after screening and cleaning to the total weight of the mixed solution is 1:2-1:

3.

7. The method for preparing the antifreeze and antibacterial frozen rotifer according to claim 5, characterized in that: The thawing method adopts a gradient thawing method, firstly placing the frozen rotifers in an environment of -10--5°C for thawing for 1-2 hours, then transferring them to an environment of 0-4°C for thawing for 30-60 minutes, and soaking the thawed rotifers in a recovery solution containing 0.1-0.2% ATP and 0.2-0.3% glutathione for 10-15 minutes before use.

8. The method for preparing the antifreeze and antibacterial frozen rotifer according to claim 5, characterized in that: The packaging adopts vacuum packaging and the packaging material is a barrier plastic film, the oxygen permeability of the film does not exceed 50cm³ / (m²・24h・0.1MPa), and the water vapor permeability does not exceed 10g / (m²・24h).

Images