CN114403308A - Fish feed additive and preparation method and application thereof - Google Patents
Fish feed additive and preparation method and application thereof Download PDFInfo
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- CN114403308A CN114403308A CN202111636850.7A CN202111636850A CN114403308A CN 114403308 A CN114403308 A CN 114403308A CN 202111636850 A CN202111636850 A CN 202111636850A CN 114403308 A CN114403308 A CN 114403308A
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- polyglutamic acid
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- polylysine
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
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- Animal Husbandry (AREA)
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- Food Science & Technology (AREA)
- Microbiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
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- Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
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Abstract
The invention discloses a fish feed additive with a growth promoting effect and application thereof, wherein the fish feed additive comprises the following components in parts by weight: the content of the polyglutamic acid is 0.2-1.0 part, the addition amount of the polylysine is 0.01-0.5 part, the addition amount of the chitosan oligosaccharide is 0.01-0.5 part, the addition amount of the bacillus subtilis is 0.01-0.5 part, and the proper addition amount is 0.01-0.5% (w/w) of the mass of the basic feed. The fish feed additive and the basic feed are stirred and fed, so that the growth of crucian can be promoted, the oxidation resistance and the digestion capability are improved, the immunity is enhanced, and the feed additive is a novel feed additive with the potential of replacing feed antibiotics. Meanwhile, the fish feed additive also has the adhesion function, can maintain the structure of the fish feed in water, improves the utilization rate of the fish feed and reduces the feed coefficient.
Description
Technical Field
The invention belongs to the field of feed additives, and relates to a fish feed additive and application thereof.
Background
According to data of '2020 Chinese fishery statistics yearbook', the total economic output value of national fishery in five years of 2015-2019 is steadily increased, wherein the fishery output value in 2019 is 12934.49 hundred million yuan, and the total economic output value is increased by 14.17% compared with that in 2015. In addition, the fresh water aquaculture yield in 2015-2019 is also stably increased from 2779 ten thousand tons in 2015 to 3013 ten thousand tons in 2019. Meanwhile, as the fishing amount of the seawater products is large, the fishing amount is reduced year by year, the yield of the seawater products is reduced, and the yield of the freshwater products is increased by 1.32 percent. However, in contrast to fishery production values and aquaculture yields, aquaculture areas and fishery practitioners have decreased year by year, with a 1.13% and 2.69% commensurate drop in 2019, respectively. The use of growth-promoting feed additives is therefore one of the important means of increasing yield. However, most of the traditional growth-promoting drug feed additives are antibiotics, so that the safety problems of serious drug residues and the like exist, and a series of national measures and reduction of culture areas and practitioners make the aquatic industry urgently need substitutes for novel growth-promoting drug feed additives.
Disclosure of Invention
The purpose of the invention is as follows: in order to solve the technical problems in the prior art, the invention provides a fish feed additive and a preparation method and application thereof.
In order to achieve the purpose, the invention provides a fish feed additive which comprises the following components in parts by weight: the content of the polyglutamic acid is 0.2-1.0 part, the addition amount of the polylysine is 0.01-0.5 part, the addition amount of the chitosan oligosaccharide is 0.01-0.5 part, and the addition amount of the bacillus subtilis is 0.01-0.5 part.
Wherein, the content of the polyglutamic acid is preferably 0.5-1.0 part, the addition amount of the polylysine is 0.01-0.2 part, the addition amount of the chitosan oligosaccharide is 0.01-0.2 part, and the addition amount of the bacillus subtilis is 0.01-0.2 part.
Preferably, the molecular weight of the polyglutamic acid is 500-2000 kDa.
The polyglutamic acid is a polyglutamic acid pure product, a polyglutamic acid fermentation liquid dried product, polyglutamic acid wettable powder or a mixture containing polyglutamic acid from natto extraction, and the dosage of the polyglutamic acid is calculated by the polyglutamic acid contained in the polyglutamic acid. The polyglutamic acid side chain contains rich carboxyl groups, can effectively crosslink organic matters and maintain the form of the feed in an aqueous solution; meanwhile, the polyglutamic acid can promote the growth of the fish, enhance the immunocompetence of the fish and the like.
The polylysine is epsilon-polylysine and a pure product of the epsilon-polylysine hydrochloride, epsilon-polylysine fermentation liquor or epsilon-polylysine fermentation liquor spray-dried powder, the polymerization degree is 9-35, preferably 15-30, and the dosage of the epsilon-polylysine is calculated by the polylysine contained in the epsilon-polylysine. Polylysine has good antibacterial property, and can be used as antibiotic substitute.
The chitosan oligosaccharide is obtained by hydrolyzing natural chitosan, the polymerization degree is 2-20, preferably 5-15, and the dosage is calculated by the chitosan oligosaccharide contained in the chitosan oligosaccharide. The chitosan oligosaccharide has good physiological activity and antiviral property, and can regulate intestinal canal morphology and improve fish absorption capacity.
The Bacillus subtilis PG-8 with the preservation number of CGMCC NO.6324 is prepared into powder for later use by spray drying after fermentation, and the effective viable count is more than or equal to 1 multiplied by 1010CFU/g. The bacillus subtilis fermentation liquor dry powder is rich in active ingredients such as antibacterial peptide, vitamins, amino acid and the like, mainly plays a nutritional role, and can promote the growth of fish, improve intestinal flora and improve the absorption capacity and the immunity of the fish.
The adding amount of the fish feed additive is 0.05-1% of the mass of the basic feed. Preferably, 0.05% -0.15% of fish feed additive and basal feed are stirred and fed to fish, so that the growth of fish can be promoted, the feed coefficient is reduced, the oxidation resistance and the digestion capacity are improved, and the immunity is enhanced.
The invention further provides application of the fish feed additive in preparation of any one or more of preparations for promoting fish growth, reducing feed coefficient, improving oxidation resistance and digestion capacity and enhancing immunity.
The fish is aquaculture fish, including but not limited to crucian, grass carp, tilapia, black carp and the like, and after the fish is used, the growth of the fish can be promoted, the feed coefficient can be reduced, the oxidation resistance and the digestion capacity can be improved, and the immunity can be enhanced.
Has the advantages that: compared with the prior art, the invention has the following advantages:
(1) according to the invention, the polyglutamic acid, the polylysine and the chitosan oligosaccharide are compounded according to the proportion to form a gel structure, so that the functions of the three components are synergized, and the granular state of the feed in a water body is maintained, the feed is not easy to disperse, and the feed utilization rate is improved;
(2) the fish feed additive and the basic feed can promote the growth of fish, reduce the feed coefficient, improve the oxidation resistance and the digestion capacity and enhance the immunity after being stirred.
Drawings
FIG. 1 is a graph of the effect of fish feed additives on the stability of fish feed in water;
FIG. 2 shows the form difference between the cultivation water tank and the carassius auratus gibelio of the G5 group and the CK group
Detailed Description
The present invention will be described in further detail with reference to specific examples, which will help understanding the present invention, but the scope of the present invention is not limited to the following examples.
Example 1 preparation of fish feed
Experimental basal feeds were purchased from premixed commercial powdered feeds from thowegian gmbh and the feed nutrients are shown in table 1.
TABLE 1 Experimental feed Nutrition ingredients
Using a blender to gradually blend 0.100% (w/w) of fish feed additives (G1, G2, G3, G4 and G5) into commercial powdered feed, meanwhile, setting the commercial powdered feed without any additive as a control group (CK), adding 1% of soybean oil and about 5% of distilled water, using an SLP-45 small feed granulator to prepare sinking granulated feed with the diameter of 2mm, and storing the sinking granulated feed in a refrigerator at-20 ℃ after being dried in the dark.
Wherein, the fish feed additive G1 comprises the following components: 5 parts of polyglutamic acid, 5 parts of polylysine, 5 parts of chitosan oligosaccharide and 5 parts of bacillus subtilis.
Wherein, the fish feed additive G2 comprises the following components: 8 parts of polyglutamic acid, 4 parts of polylysine, 4 parts of chitosan oligosaccharide and 4 parts of bacillus subtilis.
Wherein, the fish feed additive G3 comprises the following components: 11 parts of polyglutamic acid, 3 parts of polylysine, 3 parts of chitosan oligosaccharide and 3 parts of bacillus subtilis.
Wherein, the fish feed additive G4 comprises the following components: 14 parts of polyglutamic acid, 2 parts of polylysine, 2 parts of chitosan oligosaccharide and 2 parts of bacillus subtilis.
Wherein, the fish feed additive G5 comprises the following components: 16 parts of polyglutamic acid, 1 part of polylysine, 1 part of chitosan oligosaccharide and 2 parts of bacillus subtilis.
Example 2 Fish feed clumping Performance test
2g of 6 fish feeds in example 1 are weighed respectively, placed in 45mL of solution, shaken for 2min to promote the dispersion of the fish feeds in water, then kept stand for 15min, observed in the form of the feeds in the water solution, and photographed and recorded.
The experimental results are shown in fig. 1, and G4 and G5 groups significantly improved the stability of the fish feed in water.
Example 3 gelling test
To further clarify the effect of different ratios between the three soluble substances polyglutamic acid, polylysine and chitosan oligosaccharide on gelling of the mixture, we prepared 5 groups of mixtures, designated as T1, T2, T3, T4 and T5, in the proportions given in example 1, and 1% (w/v) of the solution in pure water, and then measured the storage modulus (G') and loss modulus (G ") at an angular velocity of 1rad/s using an oscillatory rheometer (TA rheometer, DHR-1, usa).
Wherein, the composition of T1 is: 5 parts of polyglutamic acid, 5 parts of polylysine and 5 parts of chitosan oligosaccharide.
Wherein, the composition of T2 is: 8 parts of polyglutamic acid, 4 parts of polylysine and 4 parts of chitosan oligosaccharide.
Wherein, the composition of T3 is: 11 parts of polyglutamic acid, 3 parts of polylysine and 3 parts of chitosan oligosaccharide.
Wherein, the composition of T4 is: 14 parts of polyglutamic acid, 2 parts of polylysine and 2 parts of chitosan oligosaccharide.
Wherein, the composition of T5 is: 16 parts of polyglutamic acid, 1 part of polylysine and 1 part of chitosan oligosaccharide.
When G 'is far greater than G', the material is mainly elastically deformed, so that the material is in a solid state; when G 'is far larger than G', the material is mainly subjected to viscous deformation, so that when the material is in a liquid state, namely G 'and G', the material is in a semi-solid state, and gel is a typical semi-solid substance.
The experimental results are shown in Table 2, wherein G 'in T4 and T5 is far greater than G', and the solution has good gel forming property and is solid. The results were in accordance with example 2.
TABLE 2 storage modulus and loss modulus of solutions of different compositions
Example 4 Effect of feed additives of the invention on the growth, antioxidant, digestive and immunological competence of Carassius auratus gibelio
The fish used in the experiment is carassius auratus gibelio, and is provided by the south spring culture base of the research center of freshwater fishery of the Chinese academy of aquatic sciences. Crucian carps with strong body and uniform size are selected in the experiment, and the initial weight is 37.26 +/-0.26 g.
The experiments were grouped as follows, with fish feed fed with the CK, G1, G2, G3, G4 and G5 fish feed additives of example 1 as experimental groups, each group being arranged in triplicate. Before the experiment, a certain amount of the allogenic silverfish are put into a storage net cage (2m multiplied by 1m), domestication is carried out for 15 days by using commercial feed, and the experiment is started after the crucian floats upwards and is ingested. Randomly selecting net cages by an experimental group, then randomly selecting 30 carassius auratus gibelio with consistent growth vigor in each feeding net cage, and recording the total weight of the carassius auratus gibelio; feeding was carried out in 3 time periods (8:00, 12:30, 17:00) per day and daily feeding was recorded. And (4) weighing, measuring the body length, sampling and the like of the fish in each net cage after 8 weeks of feeding.
After the feeding experiment is finished, fasting is carried out for 36h, 3 carassius auratus gibelio is randomly selected from each net cage, the carassius auratus gibelio is deeply anesthetized by 150mg/L MS-222, and tail vein blood collection is carried out on an ice surface by a disposable medical injector which is quickly rinsed by anticoagulant; standing the blood in an anticoagulation tube, centrifuging at 3500r/min for 10min, collecting appropriate amount of supernatant to prepare serum, and storing in a refrigerator at-20 deg.C; weighing the length of the body after blood collection, then quickly dissecting and immediately taking out the visceral mass and the liver and pancreas and weighing the visceral mass and the liver and pancreas at one time; during the period, quickly selecting about 0.1g of liver pancreas in 1mL of Trizol reagent, storing at-80 ℃ for molecular biological determination, taking a proper amount of liver pancreas for routine biochemical analysis, storing in a refrigerator at-20 ℃ for later use, taking another proper amount of liver pancreas in paraformaldehyde solution, and storing at normal temperature; simultaneously quickly stripping out an intestinal tract, taking a proper length of the crucian foregut for conventional biochemical analysis, taking about 1cm of the crucian foregut in a paraformaldehyde solution, preserving at normal temperature, taking another crucian hindgut, and carefully collecting the content of the intestinal tract; meanwhile, the back muscle of one side of the crucian is taken and stored in a refrigerator at the temperature of minus 20 ℃ for standby.
Weight gain/(%): WG ═ Wt-W0)/W0×100
Specific growth rate/(%/d): SGR ═ (ln W)t-ln W0)/t×100
The bait coefficient is as follows: FCR ═ FI/(Wt-W0)
Hepatosomatic ratio/(%): (W) HIS ═h/Wb)×100
Visceral volume ratio/(%): VSL ═ Wv/Wb)×100
Fullness/(%): CF ═ Wb/L3)×100
In the formula: w0The weight of each group of fish is equal to the weight of the fish at the end of the experiment; wtThe weight of each group of fish is equal to the weight of the fish at the end of the experiment; t is the number of days for feeding; fITotal feed intake (dry weight); whThe liver and pancreas of each fish are not heavy; wv is the visceral weight of each tail fish powder; wbThe weight of each fish powder; l is the length of each fish tail.
The moisture content of the muscle components is measured by a constant weight method (GB5009.3-2010), the crude protein content is measured by a Kjeldahl method (GB5009.5-2010), the crude fat content is measured by a Soxhlet extraction method (GB/T5009.6-2003), and the crude ash content is measured by a method (GB5009.4-2010) of burning to constant weight.
The experimental results are shown in table 3, the bait coefficient of all experimental groups is significantly reduced compared with the control group (P < 0.05); the terminal body weight was significantly higher in the G4 and G5 groups than in the CK group (P < 0.05); in the aspects of weight gain rate and specific growth rate, only the G5 group is obviously increased (P is less than 0.05) compared with the CK group, and the difference of other experimental groups is not obvious; the liver body ratio, the viscera body ratio and the fullness of the liver have little difference among the groups, and have no significant difference (P > 0.05). The water content, crude protein and ash of each experimental group were not significantly changed compared to the CK group (P > 0.05).
TABLE 3 influence of different fish feed additives on growth index and muscle composition of carassius auratus gibelio
Note IW: initial body weight; FW: terminal body weight; WG: the weight gain rate; SGR: a specific growth rate; FCR: the bait coefficient; HIS: the liver to body ratio; and VSI: the body ratio of the zang organs; CF: fullness of fertilizer
Taking a proper amount of liver and pancreas of carassius auratus gibelio, and homogenizing the mixture with normal saline according to the proportion of 1: 9 (W: V) by using a magnetic bead homogenizer in a quick ice bath to prepare 10% liver and pancreas homogenate. Then, the homogenate was divided into several equal portions, centrifuged at 3500rpm, 10000rpm and 12000rpm at 4 ℃ for 10 minutes according to the measurement of various indices, and the supernatant was stored in a refrigerator at-80 ℃ for use. The detection indexes include Malondialdehyde (MDA), superoxide dismutase (SOD), Catalase (CAT), glutathione peroxidase (GSH-Px), reduced Glutathione (GSH) and total antioxidant capacity (T-AOC), and are determined by using the kit.
The experimental results are shown in Table 4, T-AOC, CAT, SOD and GSH-Px of the G5 group are all obviously increased compared with the control group, the MDA content is obviously reduced, and the antioxidant activity of the carassius auratus gibelio is improved.
TABLE 4 influence of different fish feed additives on antioxidant capacity of liver and pancreas of Carassius auratus gibelio
Taking a proper amount of liver pancreas of carassius auratus gibelio, processing according to the above method to obtain supernatant, and determining the enzyme activities of alpha-amylase, lipase and trypsin in the liver pancreas by using a kit produced by Nanjing technology Limited.
The experimental results are shown in table 5, the digestive enzyme activity of the experimental group shows a higher level, but the enzyme activity of the G5 group is the highest, and the enzyme activities of amylase, lipase and trypsin are all significant compared with CK,
TABLE 5 Effect of different Fish feed additives on digestive enzymes in liver and pancreas of Carassius auratus gibelio
A proper amount of serum is taken, and immunoglobulin M (IgM), complement 3(C3), complement 4(C4), Glucose (GLU), Total Protein (TP), Albumin (ALB), alkaline phosphatase (ALP), Lactate Dehydrogenase (LDH), UREA (UREA), High Density Lipoprotein (HDL), Low Density Lipoprotein (LDL), Triglyceride (TG), cholesterol (TC), total bile acid (T-Bil), glutamic oxaloacetic transaminase (AST) and glutamic pyruvic transaminase (ALT) are measured by a biochemical analyzer, and related kits are all purchased from Shenzhen New industry biomedical engineering Limited.
As shown in table 6, IgM of G5, C3 and C4 all had significant increases (P <0.05) compared to the other groups, and IgM of G5 had significant increases and was not significantly different from the control group compared to the other groups. The blood glucose concentrations of G4 and G5 were significantly increased compared to the control group (P < 0.05). The total cholesterol concentration of G5 showed a significant decrease (P <0.05) compared to CK, G3 and G2, with no significant difference between the other groups. The glutamate pyruvate transaminase activities of G1 and G3 were significantly reduced compared to CK, G4 and G2 (P <0.05), while the glutamate pyruvate transaminase activity of G5 was also significant compared to CK and G4 (P < 0.05). The other indexes have no significant difference.
TABLE 6 influence of different fish feed additives on the serum biochemical index of carassius auratus gibelio
In conclusion, the group G5, i.e., the fish feed additive treatment group, has the best effects of promoting the growth of carassius auratus gibelio, improving the oxidation resistance and digestion capacity and enhancing the immunity.
The present invention provides a fish feed additive, and a method and a means for implementing the technical solution are numerous, and the above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, a plurality of modifications and embellishments can be made without departing from the principle of the present invention, and these modifications and embellishments should also be regarded as the protection scope of the present invention. All the components not specified in the present embodiment can be realized by the prior art.
Example 5 Effect of the feed additive of the invention on the growth of Tilapia mossambica
According to the results of example 4, based on the feed of group G5, 0.02%, 0.05%, 0.1%, 1%, 1.2% of fish feed additives were prepared according to the method described in example 1 and named G5-0.02, G5-0.05, G5-0.1, G5-1, G5-1.2, respectively.
The fish used in the experiment is the gift tilapia, and is provided by the south spring aquaculture base of the research center of freshwater fishery of the Chinese academy of aquatic sciences. Tilapia mossambica with robust fish bodies and uniform size is selected in the experiment, and the initial weight is 31.48 +/-3.16 g.
The experiment groups are as follows, the fish feed which is fed with the fish feed additive G5-0.02, G5-0.05, G5-0.1, G5-1 and G5-1.2 is taken as an experiment group, and three groups are arranged in parallel. Before the experiment begins, a certain amount of tilapia is put into a storage net cage (2m multiplied by 1m), domestication is carried out for 15 days by using commercial feed, and the experiment begins after the tilapia floats upwards and ingests food. Randomly selecting net cages by an experimental group, then randomly selecting 30 tilapia mossambica with consistent growth vigor in each feeding net cage, and recording the total weight of the tilapia mossambica; feeding was carried out in 3 time periods (8:00, 12:30, 17:00) per day and daily feeding was recorded. And weighing, measuring the body length, sampling and the like are carried out on the fish in each net cage after the fish is raised for 6 weeks.
The experimental results are shown in Table 7, the promotion effect of the G5-0.1 treatment group on the growth of tilapia is most obvious, and the effects of the G5-0.05 and G5-1 treatment groups are also obvious. The G5-0.02 and G5-1.2 treated groups were similar to the control group in effect and had no significant growth promoting effect.
TABLE 7 Effect of different amounts of added fish feed additives on the growth of Tilapia mossambica
Note: FW: terminal body weight; WG: the weight gain rate; FCR: and (4) bait coefficient.
Claims (9)
1. The fish feed additive is characterized by comprising the following components in parts by weight: the content of the polyglutamic acid is 0.2-1.0 part, the addition amount of the polylysine is 0.01-0.5 part, the addition amount of the chitosan oligosaccharide is 0.01-0.5 part, and the addition amount of the bacillus subtilis is 0.01-0.5 part.
2. The fish feed additive according to claim 1, wherein the polyglutamic acid is preferably 0.5-1.0 part, the polylysine is 0.01-0.2 part, the chitosan oligosaccharide is 0.01-0.2 part, and the bacillus subtilis is 0.01-0.2 part.
3. The fish feed additive according to claim 1, wherein the polyglutamic acid has a molecular weight of 500-2000 kDa, and is a polyglutamic acid-derived mixture derived from pure polyglutamic acid, pure polyglutamate, polyglutamic acid fermentation broth, dried polyglutamic acid fermentation broth, wettable polyglutamic acid powder, or natto, in terms of the amount of polyglutamic acid contained therein.
4. The fish feed additive according to claim 1, wherein the polylysine is epsilon-polylysine and a pure product of hydrochloride thereof, epsilon-polylysine fermentation broth or epsilon-polylysine fermentation broth spray-dried powder, the polymerization degree is 9-35, and the dosage is calculated by the polylysine contained therein.
5. The fish feed additive according to claim 1, wherein the chitosan oligosaccharide is obtained by hydrolyzing natural chitosan, and has a degree of polymerization of 2 to 20, in terms of the amount of chitosan oligosaccharide contained therein.
6. The fish feed additive of claim 1, wherein the Bacillus subtilis PG-8 with the preservation number of CGMCC number 6324 is prepared into powder for later use by spray drying after fermentation, and the effective viable count is more than or equal to 1 x 1010 CFU/g。
7. The fish feed additive of claim 1, wherein the amount of the fish feed additive added is 0.05-1% by mass of the basal feed.
8. Use of a fish feed additive according to claim 1 in the preparation of a formulation for any one or more of promoting fish growth, reducing feed coefficient, increasing antioxidant and digestive ability, and enhancing immune ability.
9. Use according to claim 8, wherein the fish is an aquaculture fish.
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| CN116267713A (en) * | 2023-02-27 | 2023-06-23 | 南方海洋科学与工程广东省实验室(珠海) | Method for breeding groupers with fresh and tender meat quality |
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