CN104719639A - Method for preparing antibacterial feed protein by utilizing aquatic product leftovers - Google Patents
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Abstract
The invention discloses a method for preparing antibacterial protein by utilizing aquatic product leftovers. The method comprises the following main steps: mincing the fresh aquatic product leftovers for later use, crushing vegetable feed proteins, and filtering the crushed vegetable feed proteins by virtue of a 60-mesh screen for later use; mixing the minced aquatic product leftovers and the crushed vegetable proteins, and uniformly mixing nutrients and water to be used as a fermenting culture base; sterilizing and cooling the fermenting culture base, placing bacillus subtilis HL-1 capable of generating subtilin, and carrying out the solid aerobic fermentation; drying, crushing and packaging the fermented product to obtain the antibactieral feed protein. The prepared antibacterial feed protein is rich in subtilin, nutritional small peptide and probiotics and has a good function of promoting the growth and resisting the diseases. By adopting the antibacterial feed protein, not only can the utilization rate of the protein be increased, but also the antimicrobial-free feed can be prepared by utilizing the antibacterial feed protein, and the application prospect and the social benefit are good.
Description
Technical field
The present invention relates to a kind of method utilizing aquatic product leftovers to prepare antiseptic feed albumen, belong to processing of aquatic products and utilize field.
Background technology
There are 9.7 ten thousand tons for livestock breeding industry in about 210,000 tons of antibiotic raw materials that current China produces every year, account for gross annual output amount 46.1%.Excessive use antibiotic not only result in the drug resistance of cultivated animals to antibiotic medicine, the resistance against diseases of cultivated animals is declined, and these antibiotic can enter human body with food chain, human body is made inadvertently to produce drug-resistant bacteria, treatment difficulty and expense will be increased once sick, even likely can produce " superbacteria ", occur situation about pasting medical help.Along with improving constantly of people's quality of life, safe and sanitary varieties of food items being comprised to meat product has had higher requirement, also day by day strong to the cry of " quality-assured meat ", " pollution-free food ".Therefore, finding antibiotic green substitute is China's feed and problem demanding prompt solution in cultivation industry development.
As a kind of substitute of feeding antibiotic, antibacterial peptide usually has heat-resisting, has a broad antifungal spectrum, good water solubility and not easily produces the characteristics such as drug resistance, receives much concern in recent years.Antibacterial peptide derive from animal, plant and microorganism, but animal and plant source antibacterial peptide is due to limited source, and application is restricted; And microbial antibacterial peptide can large-scale production, advantage of lower cost, thus microbial source antibacterial peptide more has application prospect.Current microbial source antibacterial peptide most study be the antibacterial peptide of lactic acid bacteria, and in the world to bacillus subtilis (
bacillus subtilis) this novel fine bacteriums of the subtilin antibacterial peptide research that produces is then little, domestic rare especially.Industrial microorganism (2008,38(4): 14 ~ 18) disclose a kind of bacillus subtilis (
bacillus subtilis) FB123 produces the liquid fermentation condition of subtilin, but the solid state fermentation of subtilin is produced also not studies have reported that.
Within 1988, nisin obtains the accreditation of FDA first as food additives, has and is using nisin as food preservative more than 100 countries and regions.The direct use of antibacterial peptide on food, it is safe for this also illustrates antibacterial peptide to animals and humans.But at present, because high production cost makes the application of antibacterial peptide mainly at field of food, and little in the application of feedstuff industry.Therefore the production cost reducing antibacterial peptide is badly in need of, to promote that antibacterial peptide is in the application of feed industry.
Solid fermentation has the advantages that investment is low and operating cost is low; And tunning convection drying and pulverizing are prepared into finished product, greatly reduce post processing cost.Thus, compared with liquid fermentation, solid fermentation produces the production cost that antibacterial peptide obviously can reduce antibacterial peptide.Patent CN101381757A " the solid fermentation method preparations and applicatio of antibacterial peptide " discloses a kind of solid fermentation method of antibacterial peptide.Agricultural University Of Nanjing's Master's thesis in 2010 "
bacillus amyloliquefaciesthe research of ES-2-4 antibacterial lipopeptid solid-state fermentation process " disclose a kind of solid state fermentation production technology of bacillus amyloliquefaciens antibacterial lipopeptid.Although solid state fermentation produces the production cost that antibacterial peptide can reduce antibacterial peptide greatly, there be limited evidence currently of has the antibacterial peptide products application of solid state fermentation in feed industry.
Summary of the invention
Although the antibacterial peptide finished product that solid fermentation method is produced can reduce the production cost of antibacterial peptide, because content is low, the antibacterial peptide finished product of low dosage can not play effective effect in feed; The antibacterial peptide finished product of high dose affects again the formula Design of feed.For this problem that the antibacterial peptide product of current solid fermentation exists, the object of the invention is to be prepared into a kind of antiseptic feed albumen, with the antibacterial peptide series products significantly increasing solid fermentation addition and don't affect the design of feed formula in mixed feed.
In order to solve the problem, the technical solution adopted in the present invention is as follows: a kind of method utilizing aquatic product leftovers to prepare antiseptic feed albumen, comprises the steps:
(1) by slant preservation can produce subtilin bacillus subtilis (
bacillus subtilis) in the LB fluid nutrient medium that cools after being linked into sterilizing of HL-1, in 37 DEG C, 150r/min cultivates 16h, be prepared into bacillus subtilis (
bacillus subtilis) HL-1 seed liquor is for subsequent use;
(2) for subsequent use after fresh aquatic product leftovers being rubbed, cross 60 mesh sieves after being pulverized by vegetable feed proteins for subsequent use;
(3) aquatic product leftovers of rubbing step (2) obtained and the rear vegetable feed proteins crossing 60 mesh sieves of pulverizing are that the ratio of 0.5 ~ 1.5:1 mixes in mass ratio, and Ensure Liquid thing and water mixing, obtain final moisture content and control the fermentation medium in 40% ~ 80% scope;
(4) fermented and cultured of mixing step (3) obtained is based on 121 DEG C of sterilizing 30min;
(5) fermentation medium after sterilizing step (4) obtained is cooled to room temperature, then access bacillus subtilis prepared by step (1) (
bacillus subtilis) HL-1 seed liquor, and mix;
(6) fermentation of raw material: the fermentation medium adding bacterium that step (5) obtains is fermented according to following method: fermentation medium is divided in the fermentation tray of having sterilized, charging thickness is 4cm ~ 8cm, the sterilized wet gauze of fermentation medium surface coverage in tray, again tray is placed in the shelf top fermentation in sterilized solid fermentation room, the temperature that controls environment is at 28 DEG C ~ 30 DEG C, and fermentation time is 35h ~ 45h;
(7) product handling system: fermentation ends, by the product drying obtained after fermentation, after pulverizing and packaging, has namely obtained a kind of antiseptic feed albumen finished product utilizing aquatic product leftovers to prepare.
The bacillus subtilis of access in described step (1) (
bacillus subtilis) HL-1 can produce subtilin, this bacterium is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on January 20th, 2015, deposit number is CGMCC No.10379, and preservation address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica.
Aquatic product leftovers in described step (2) is the one in leftovers of tilapia, shrimp head, scallop body, cod leftover bits and pieces, squid leftover bits and pieces, carp leftover bits and pieces, Harengula zunasi leftover bits and pieces, black carp leftover bits and pieces, silver carp leftover bits and pieces, grass carp leftover bits and pieces and stripped tuna leftover bits and pieces; Vegetable feed proteins in described step (2) is a kind of or wherein two or more composition in dregs of beans, peanut meal, Cottonseed Meal and the dish dregs of rice.。
The mixture that nutrients in described step (3) is glucose, frerrous chloride and dipotassium hydrogen phosphate form; Nutraceutical addition in described step (3) accounts for fermentation medium gross mass percentages with the nutraceutical quality of adding, and is respectively: glucose 0.01% ~ 5%, frerrous chloride 0.01% ~ 5%, dipotassium hydrogen phosphate 0.01% ~ 5%.
The middle bacillus subtilis of described step (5) (
bacillus subtilis) inoculum concentration of HL-1 seed liquor accounts for fermentation medium gross mass percentages with the quality accessing seed liquor, is 0.1% ~ 5%.
The present invention compares with existing technology, has the following advantages.
(1) antibacterial peptide product protein content is high, can carry out formula Design neatly for feed industry.The protein content of antiseptic feed albumen prepared by the present invention is greater than 50%, and the protein content of this content and three grades of fish meal maintains an equal level.Antiseptic feed albumen prepared by the present invention addition in prawn mixed feed even can reach more than 20%, and does not affect the design of prawn compound feed prescription.
(2) the little peptide of nutrition is rich in.The present invention is high due to fermentation medium protein content, can a large amount of little peptide of nutrition in sweat, and this not only can improve the utilization rate of protein, and also has obvious growth-promoting effect.
The invention provides a kind of method utilizing aquatic product leftovers to prepare antiseptic feed albumen.The culture experiment of the pig of antiseptic feed albumen in South China that the present invention obtains, chicken, prawn shows, the feed that the antiseptic feed albumen produced with the present invention coordinates, not only can improve the growth result of letting animals feed, but also immunity and the premunition of letting animals feed can be improved, directly can be used for preparing nonreactive feed, have a good application prospect and social effect.
Detailed description of the invention
In order to understand the present invention better, with regard to the selection of the kind of aquatic product leftovers and vegetable feed proteins, fermentation parameter and fermentation mode, the invention will be further described in conjunction with the embodiments below, but protection scope of the present invention is not limited thereto.
Embodiment 1: by slant preservation can produce subtilin bacillus subtilis (
bacillus subtilis) in the LB fluid nutrient medium that cools after being linked into sterilizing of HL-1, in 37 DEG C, 150r/min cultivates 16h, be prepared into bacillus subtilis (
bacillus subtilis) HL-1 seed liquor is for subsequent use.It is for subsequent use after fresh leftovers of tilapia is rubbed; 60 mesh sieves are crossed for subsequent use after being pulverized by dregs of beans.By 100 kilograms rub leftovers of tilapia and 100 kilograms pulverizes and crosses 60 mesh sieves dregs of beans mixing, and add 400 grams of glucose, 200 grams of frerrous chlorides, 500 grams of dipotassium hydrogen phosphates mix together with 10 kg of water, acquisition mix after fermentation medium; Fermented and cultured after mixing is based on 121 DEG C of sterilizing 30min; Then add in the fermentation medium cooled after sterilizing 2 kilograms of bacillus subtilises (
bacillus subtilis) HL-1 seed liquor, stir and mix; Postvaccinal fermentation medium is divided in the fermentation tray of the alcohol surface sterilization using 75%, thickness of feeding in tray is 8 centimetres, the wet gauze of fermentation medium surface coverage sterilization treatment in tray, again tray is placed in the shelf top fermentation in the solid fermentation room of sterilizing, the temperature that controls environment is 28 DEG C, and fermentation time is 45h; After fermentation ends, by the product drying obtained after fermentation, after pulverizing and packaging, namely obtain a kind of antiseptic feed albumen finished product utilizing aquatic product leftovers to prepare of the present invention.
Embodiment 2: by slant preservation can produce subtilin bacillus subtilis (
bacillus subtilis) in the LB fluid nutrient medium that cools after being linked into sterilizing of HL-1, in 37 DEG C, 150r/min cultivates 16h, be prepared into bacillus subtilis (
bacillus subtilis) HL-1 seed liquor is for subsequent use.It is for subsequent use after fresh shrimp head is rubbed; By for subsequent use for broken for peanut meal rear mistake 60 mesh sieve.By 80 kilograms rub shrimp heads and 100 kilograms pulverizes and crosses 60 mesh sieves peanut meal mixing, and add 300 grams of glucose, 200 grams of frerrous chlorides, 450 grams of dipotassium hydrogen phosphates mix together with 10 kg of water, acquisition mix after fermentation medium; Fermented and cultured after mixing is based on 121 DEG C of sterilizing 30min; Then add in the fermentation medium cooled after sterilizing 2 kilograms of bacillus subtilises (
bacillus subtilis) HL-1 seed liquor, stir and mix; Postvaccinal raw material is divided in the fermentation tray of the alcohol surface sterilization using 75%, thickness of feeding in tray is 4 centimetres, the wet gauze of material surface coverage sterilization treatment in tray, again tray is placed in the shelf top fermentation in the solid fermentation room of sterilizing, the temperature that controls environment is 30 DEG C, and fermentation time is 35h; After fermentation ends, by the product drying obtained after fermentation, after pulverizing and packaging, namely obtain a kind of antiseptic feed albumen finished product utilizing aquatic product leftovers to prepare of the present invention.
Embodiment 3: by slant preservation can produce subtilin bacillus subtilis (
bacillus subtilis) in the LB fluid nutrient medium that cools after being linked into sterilizing of HL-1, in 37 DEG C, 150r/min cultivates 16h, be prepared into bacillus subtilis (
bacillus subtilis) HL-1 seed liquor is for subsequent use.It is for subsequent use after scallop body is rubbed; 60 mesh sieves are crossed for subsequent use after being pulverized by Cottonseed Meal.By 120 kg rub scallop body and 100 kilograms pulverizes and crosses 60 mesh sieves Cottonseed Meal mixing, and add 250 grams of glucose, 150 grams of frerrous chlorides, 400 grams of dipotassium hydrogen phosphates mix together with 5 kg of water, acquisition mix after fermentation medium; Mixed fermented and cultured is based on 121 DEG C of sterilizing 30min; Then add in the fermentation medium cooled after sterilizing 2 kilograms of bacillus subtilises (
bacillus subtilis) HL-1 seed liquor, stir and mix; Postvaccinal raw material is divided in the fermentation tray of the alcohol surface sterilization using 75%, thickness of feeding in tray is 6 centimetres, the wet gauze of material surface coverage sterilization treatment in tray, again tray is placed in the shelf top fermentation in the solid fermentation room of sterilizing, the temperature that controls environment is at 28 DEG C, and fermentation time is 43h; After fermentation ends, by the product drying obtained after fermentation, after pulverizing and packaging, namely obtain a kind of antiseptic feed albumen finished product utilizing aquatic product leftovers to prepare of the present invention.
Embodiment 4: by slant preservation can produce subtilin bacillus subtilis (
bacillus subtilis) in the LB fluid nutrient medium that cools after being linked into sterilizing of HL-1, in 37 DEG C, 150r/min cultivates 16h, be prepared into bacillus subtilis (
bacillus subtilis) HL-1 seed liquor is for subsequent use.It is for subsequent use after cod leftover bits and pieces is rubbed; 60 mesh sieves are crossed for subsequent use after being pulverized by the dish dregs of rice.By 120 kg rub after cod leftover bits and pieces and 100 kilograms pulverizes and crosses 60 mesh sieves the dish dregs of rice mixing, and add 350 grams of glucose, 120 grams of frerrous chlorides, 300 grams of dipotassium hydrogen phosphates mix together with 5 kg of water, acquisition mix after fermentation medium; Mixed fermented and cultured is based on 121 DEG C of sterilizing 30min; Then add in the fermentation medium cooled after sterilizing 2 kilograms of bacillus subtilises (
bacillus subtilis) HL-1 seed liquor, stir and mix; Postvaccinal raw material is divided in the fermentation tray of the alcohol surface sterilization using 75%, thickness of feeding in tray is 8 centimetres, the wet gauze of material surface coverage sterilization treatment in tray, again tray is placed in the shelf top fermentation in the solid fermentation room of sterilizing, the temperature that controls environment is at 28 DEG C, and fermentation time is 45h; After fermentation ends, by the product drying obtained after fermentation, after pulverizing and packaging, namely obtain a kind of antiseptic feed albumen finished product utilizing aquatic product leftovers to prepare of the present invention.
Embodiment 5: by slant preservation can produce subtilin bacillus subtilis (
bacillus subtilis) in the LB fluid nutrient medium that cools after being linked into sterilizing of HL-1, in 37 DEG C, 150r/min cultivates 16h, be prepared into bacillus subtilis (
bacillus subtilis) HL-1 seed liquor is for subsequent use.It is for subsequent use after Harengula zunasi leftover bits and pieces is rubbed; By peanut meal and broken rear mistake 60 mesh sieve of Cottonseed Meal for subsequent use.By 80 kilograms rub after Harengula zunasi leftover bits and pieces and 100 kilograms pulverizes and cross 60 mesh sieves mixed feed albumen (50 kilograms of peanut meals+50 kilograms of Cottonseed Meals) mix, and add 400 grams of glucose, 150 grams of frerrous chlorides, 400 grams of dipotassium hydrogen phosphates mix together with 10 kg of water, obtain mixing after fermentation medium; Mixed fermented and cultured is based on 121 DEG C of sterilizing 30min; Then add in the fermentation medium cooled after sterilizing 1 kilogram of bacillus subtilis (
bacillus subtilis) HL-1 seed liquor, stir and mix; Postvaccinal raw material is divided in the fermentation tray of the alcohol surface sterilization using 75%, thickness of feeding in tray is 4 centimetres, the wet gauze of material surface coverage sterilization treatment in tray, again tray is placed in the shelf top fermentation in the solid fermentation room of sterilizing, the temperature that controls environment is at 29 DEG C, and fermentation time is 43h; After fermentation ends, by the product drying obtained after fermentation, after pulverizing and packaging, namely obtain a kind of antiseptic feed albumen finished product utilizing aquatic product leftovers to prepare of the present invention.
Claims (7)
1. utilize aquatic product leftovers to prepare a method for antiseptic feed albumen, it is characterized in that: the method comprises the steps:
(1) in the LB fluid nutrient medium cooled after the bacillus subtilis HL-1 that can produce subtilin of slant preservation being linked into sterilizing, in 37 DEG C, 150r/min cultivates 16h, is prepared into bacillus subtilis HL-1 seed liquor for subsequent use;
(2) for subsequent use after fresh aquatic product leftovers being rubbed, cross 60 mesh sieves after being pulverized by vegetable feed proteins for subsequent use;
(3) aquatic product leftovers of rubbing step (2) obtained and the rear vegetable feed proteins crossing 60 mesh sieves of pulverizing are that the ratio of 0.5 ~ 1.5:1 mixes in mass ratio, and Ensure Liquid thing and water mixing, obtain final moisture content and control the fermentation medium in 40% ~ 80% scope;
(4) fermented and cultured of mixing step (3) obtained is based on 121 DEG C of sterilizing 30min;
(5) fermentation medium after sterilizing step (4) obtained is cooled to room temperature, then accesses the bacillus subtilis HL-1 seed liquor prepared by step (1), and mixes;
(6) fermentation of raw material: the fermentation medium adding bacterium that step (5) obtains is fermented according to following method: fermentation medium is divided in the fermentation tray of having sterilized, charging thickness is 4 ~ 8cm, the sterilized wet gauze of fermentation medium surface coverage in tray, again tray is placed in the shelf top fermentation in sterilized solid fermentation room, the temperature that controls environment is at 28 DEG C ~ 30 DEG C, and fermentation time is 35h ~ 45h;
(7) product handling system: fermentation ends, by the product drying obtained after fermentation, after pulverizing and packaging, has namely obtained a kind of antiseptic feed albumen finished product utilizing aquatic product leftovers to prepare.
2. a kind of method utilizing aquatic product leftovers to prepare antiseptic feed albumen according to claim 1, is characterized in that: bacillus subtilis HL-1 used in described step (1) is CGMCC No.10379 at the deposit number at China Committee for Culture Collection of Microorganisms's common micro-organisms center.
3. a kind of method utilizing aquatic product leftovers to prepare antiseptic feed albumen according to claim 1, is characterized in that: the aquatic product leftovers in described step (2) is the one in leftovers of tilapia, shrimp head, scallop body, cod leftover bits and pieces, squid leftover bits and pieces, carp leftover bits and pieces, Harengula zunasi leftover bits and pieces, black carp leftover bits and pieces, silver carp leftover bits and pieces, grass carp leftover bits and pieces and stripped tuna leftover bits and pieces.
4. a kind of method utilizing aquatic product leftovers to prepare antiseptic feed albumen according to claim 1, is characterized in that: the vegetable feed proteins in described step (2) is a kind of or wherein two or more mixture in dregs of beans, peanut meal, Cottonseed Meal and the dish dregs of rice.
5. a kind of method utilizing aquatic product leftovers to prepare antiseptic feed albumen according to claim 1, is characterized in that: the mixture that the nutrients in described step (3) is glucose, frerrous chloride and dipotassium hydrogen phosphate form.
6. a kind of method utilizing aquatic product leftovers to prepare antiseptic feed albumen according to claim 1, it is characterized in that: the nutraceutical addition in described step (3) accounts for fermentation medium gross mass percentages with the nutraceutical quality of adding, be respectively: glucose 0.01% ~ 5%, frerrous chloride 0.01% ~ 5%, dipotassium hydrogen phosphate 0.01% ~ 5%.
7. a kind of method utilizing aquatic product leftovers to prepare antiseptic feed albumen according to claim 1, it is characterized in that: in described step (5), the inoculum concentration of bacillus subtilis HL-1 seed liquor accounts for fermentation medium gross mass percentages with the quality accessing seed liquor, is 0.1% ~ 5%.
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| CN105685478A (en) * | 2016-03-09 | 2016-06-22 | 广东海洋大学 | Method for preparing green growth-promoting feed additive through high-temperature fermentation of aquatic protein |
| CN105685479A (en) * | 2016-03-09 | 2016-06-22 | 广东海洋大学 | Preparation method of fermented soybean meal |
| CN105724764A (en) * | 2016-03-09 | 2016-07-06 | 广东海洋大学 | Method for preparing green antibacterial growth-promoting feed additive by utilizing aquatic protein fermentation |
| CN105746890A (en) * | 2016-03-09 | 2016-07-13 | 广东海洋大学 | Method of utilizing fermentation of aquatic product protein and plant feed protein to prepare fish meal replacing protein |
| CN105746892A (en) * | 2016-03-09 | 2016-07-13 | 广东海洋大学 | Method of utilizing solid-state fermentation of aquatic product protein to prepare green growth-promoting feed additive |
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| CN106689795A (en) * | 2015-11-13 | 2017-05-24 | 江苏银宝生物科技有限公司 | Feed formula for artificially breeding saltwater fish and processing method of feed formula |
| CN107173521A (en) * | 2016-03-09 | 2017-09-19 | 广东海洋大学 | A kind of preparation method of ferment feather powder |
| CN107173534A (en) * | 2016-03-09 | 2017-09-19 | 广东海洋大学 | A kind of utilization aquatic product protein and feather meal fermentation prepare the method that fish meal substitutes albumen |
| CN107319135A (en) * | 2017-07-27 | 2017-11-07 | 珠海天凯生物科技有限公司 | A kind of premixed feed and preparation method thereof |
| CN109170130A (en) * | 2018-08-08 | 2019-01-11 | 浙江炯炜海洋生物科技有限公司 | The method that solid state fermentation aquatic products leftover bits and pieces prepares fish meal |
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| CN106689795A (en) * | 2015-11-13 | 2017-05-24 | 江苏银宝生物科技有限公司 | Feed formula for artificially breeding saltwater fish and processing method of feed formula |
| CN107173521A (en) * | 2016-03-09 | 2017-09-19 | 广东海洋大学 | A kind of preparation method of ferment feather powder |
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| CN105685479A (en) * | 2016-03-09 | 2016-06-22 | 广东海洋大学 | Preparation method of fermented soybean meal |
| CN107173534A (en) * | 2016-03-09 | 2017-09-19 | 广东海洋大学 | A kind of utilization aquatic product protein and feather meal fermentation prepare the method that fish meal substitutes albumen |
| CN105746892B (en) * | 2016-03-09 | 2019-10-29 | 广东海洋大学 | A method of green feed additive for promoting growth is prepared using aquatic product protein solid state fermentation |
| CN105746890A (en) * | 2016-03-09 | 2016-07-13 | 广东海洋大学 | Method of utilizing fermentation of aquatic product protein and plant feed protein to prepare fish meal replacing protein |
| CN105685478B (en) * | 2016-03-09 | 2019-09-10 | 广东海洋大学 | A method of green feed additive for promoting growth is prepared using aquatic product protein hot fermentation |
| CN105918674A (en) * | 2016-05-16 | 2016-09-07 | 栾文华 | Composite premix for sows in perinatal period |
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| CN109170130B (en) * | 2018-08-08 | 2022-02-08 | 浙江炯炜海洋生物科技有限公司 | Method for preparing fish meal by solid-state fermentation of aquatic product leftovers |
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