CN103027187A - Anti-stress fermentation protein feed and producing method thereof - Google Patents

Anti-stress fermentation protein feed and producing method thereof Download PDF

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CN103027187A
CN103027187A CN2013100016255A CN201310001625A CN103027187A CN 103027187 A CN103027187 A CN 103027187A CN 2013100016255 A CN2013100016255 A CN 2013100016255A CN 201310001625 A CN201310001625 A CN 201310001625A CN 103027187 A CN103027187 A CN 103027187A
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fermentation
stress
feed
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dregs
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黄谦
于家伊
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BEIJING JIABOWEN BIOSCIENCE Co Ltd
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BEIJING JIABOWEN BIOSCIENCE Co Ltd
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Abstract

The invention provides an anti-stress fermentation protein feed which is produced by composite bacterium inoculation and high-temperature and moderate-temperature fermentation of a plant derived protein raw material, a terrestrial animal derived protein raw material and a marine animal derived protein raw material. Macromolecular substances in the protein feed are digested and decomposed into micromolecular substances, stressors in nutritional ingredients are decreased, anti-stress active factors such as genistein, glutathione and glucosamine are increased, the immunity of fed animals is improved, and the protein feed shows a good anti-stress capability. The invention further provides a producing method of the anti-stress fermentation protein feed. The feed product with high functionality, good anti-stress effect and high economical efficiency is obtained with the adoption of the multiple derived protein raw materials, a specific formula and an appropriate matching ratio and by mixed fermentation through a special secondary fermentation process.

Description

Resisting stress fermentation protein feedstuff and production method thereof
 
Technical field
The present invention relates to a kind of multielement protein composition of raw materials and by secondary fermentation produce anti-stress the fermentation protein concentrated feed.
 
Background technology
Through the practice of recent two decades, intensive and industrialized aquaculture has become the Main Patterns of aquatic products, livestock and poultry cultivation.This pattern also seriously exposes the problem that this system can't overcome that exists---Animal stress syndrome behind when having brought revolutionary variation to aquaculture, it has become the most active factor of obstacle aquaculture development.
Canadian pathologic, physiologic scholar Selye proposed theory of stress in 1936, stress be the factor overstimulations such as the body biology that is subjected to exist in extraneous and the internal environment, physics and chemistry after, the immediately a series of non-specific whole body adaptation reaction of generation.These stress reactions in various degree, the normal growth that not only can directly affect animal is grown, and the decline of serious caused immunity of organisms, brings out ill even dead.
The generation of stress reaction at first is owing to be subject to the stimulation of various stressors.In the animal-breeding growth course, often suffer vaccine inoculation, use medicine, resettlement, extremely hot, the abrupt change of climate etc. stress.By stress causing just feed intake decline, amino acid, vitamin, electrolyte absorption to reduce, and synthesis capability descends in the body, causes a series of variation of nutrition, physiology, metabolism.Stress can not be ignored to the loss that Production of Livestock and Poultry causes.Along with animal production scale and industrialization, the cultivation intensive degree is more and more higher, and animal is subjected to more and more stress; causing stress be excessive; be animal body when being subject to long-time or high-intensity stressor and stimulating, will produce serious adverse effect, thus the harm body.What cause thus stress often cause catabolism enhancing in the animal body, anabolism reduces, show as growth retardation, Body weight loss, feed conversion rate reduction, product quality decline, immunity of organisms reduction, more and more have influence on aborning animal the production achievement (Yang Yufeng etc., anti-stress the application of additive on Production of Livestock and Poultry.Today animal and veterinary, 08 phase in 2006).
In early days to stress the research of mechanism mainly concentrate on nerve and internal system, finding afterwards stress be also closely related with body immune system.Strong stress stimulation can cause nerve, endocrinopathy, immune dysfunction, aggregate performance is that the quantity of the immunocytes such as T, B lymphocyte, NK cell reduces, proliferation activity lowers, special and nonspecific immune reaction ability weakens, immune-regulating factor (IL-2, IFN etc.) produces and reduces (Tracy BH, Sheldon C. Stress and immunity in humans: a meta analytic review. Psychosomatic Medicine, 1993,55:364-379.).Be the excitement of hypothalamus-hypophysis-adrenal cortex axle system (HPA ax2is) and sympathetic nerve-adrenal medella system at the initial stage of stress reaction main manifestations, glucocorticoid secretion increases, and immune dysfunction is neural and the direct result of endocrine reaction, middle and later periods then shows as stress disorders, immunologic hypofunction (Eskandari F, Sternberg EM. Neural immune interactions in health and disease. Ann N Y Acad Sci, 2002,966:20-27.).Dissimilar stress may have different to the impact of immunologic function, but on the impact of immunologic function take cell immune response as main, show as the T lymphocyte quantity and reduce, proliferation activity reduces, immunological regulation and kill capability descend.The humoral immune reaction relative complex; relevant with stressor kind, intensity, duration etc.; such as the early metaphase stage at heat stress; the B lymphocyte antibody produces and increases; then then showing as cytoactive descends; antibody produces inhibition, and (Jin Hanghong etc., physiology and psychological stress reaction are on impact and the safeguard measure of immunologic function.China's Modern Nursing magazine, 2007(4), 7:591-594).
So, stress affect Immune Function In Animals by increasing blood plasma cortisol, reduce phagocytic function and suppressing antibody response.In addition, interior free yl is increased, further affect the content of vitamin E in the body, (Li Liuan etc., transport stress is to the progress of animal body function effect to have reduced the resistance against diseases of body.HEILONGJIANG ANIMAL SCIENCE AND VETERINARY MEDICINE, science and technology version, 2010,9:40-41).Immunity of organisms descends, premunition weakens, and easily causes infectious disease and epiphytotics popular.Stress be the source of ten thousand diseases thus.
Although at present correspondence excites living control method a lot, to alleviate or control stress measure still very imperfect.So obtain good anti-stress effect, should understand fully as far as possible stressor, take comprehensive measure (Antistress agent that reinforcement feeding and management, reasonable preparation daily ration, use have no side effect etc.), (Yang Yufeng etc., resisting stress the application of additive on Production of Livestock and Poultry just to obtain satisfied effect.Today animal and veterinary, 08 phase in 2006).
Anti-stress factor refers to have alleviation, control by stressor cause stress syndrome material, anti-stress the mechanism of action also have nothing in common with each other, sum up, mainly contain several aspects: 1. regulate central nervous system, play stable calm, reduction stress sensitive effect.As by nutrition such as tryptophan, tyrosine, taurine, 5-OH tryptamines, Chinese herbal medicine such as radix bupleuri etc. affects the secretion of hypothalamus cortex hormone of aadrenaline releasing hormone; Turn down accretion rate, prevent that metabolism is excessively to the damage of body.2. anti-oxidant, Scavenger of ROS free radical, regulate by the Radical Metabolism that stress cause disorderlyly, be many anti-major ways that stress additive play a role, such as VC, VE, glutathione, astaxanthin etc.); 3. regulate the various metabolisms of body, regulate unusual by the organism metabolism that stress cause; 4. improve immunity of organisms, strengthen resistance against diseases.
The below introduces several related to the present invention, can improve body immunity, has anti-oxidant, as to remove Scavenging Oxygen Free Radical anti-stress factor.
The natural tripeptides that contains sulfydryl that glutathione (glutathione, GSH) is comprised of glutamic acid, cysteine and glycine, as ubiquitous small molecule antioxidant in the eukaryotic, the oxygen radical in body is eliminated and is played a crucial role.
Glutathione easily with the combinations such as some drugs, toxin (such as free radical, iodoacetic acid, mustard gas, heavy metals such as lead, mercury, arsenic), have antioxidation and integration detoxication.Suitable redox environment plays vital effect (Izawa S for keeping body in vivo, etal. Oxidative stress response in yeast:effect of glutathione on adaptation to hydrogen peroxide stress in Saccharomyces cerevisiae. FEBS Lett. 1995, Jul, 10:368 (1): 73-6.; Zhu Xuan etc., dietary glutathione organize the impact of GSH-PX activity deposition and oxidation resistance on grass carp.Chinese aquatic science, 2008,01:160-166).Studies show that GSH can improve oxidation resistance and the digestive enzyme activity of aquatic livestock, excitating organism immunity improves anti-stress ability, thereby promotes body growth (Zambohino Infante J. etal; Partial substitution of diand tripeptides for native proteins in sea bass diets improve dicentrarchus labras larval development. J Nutr., 1997 Apr; 127 (4): 608-14; Liu Xiaohua etc., dietary glutathione is on the impact of Environment of Litopenaeus vannamei Low hepatopancrease Antioxidant Indexes and lipid peroxide contents.Aquatic product journal, 2007,02:235-240; Zhao Hongxia etc., dietary glutathione is on the impact of Growth of Grass Carps Ctenopharyngodon Idellus, physical signs and premunition.Chinese aquatic science, 2007,04:678-683; Cao Junming etc., dietary glutathione is on the impact of the Environment of Litopenaeus vannamei Low nospecific immunity factor and related enzyme activity.Hua Zhong Agriculture University's journal, 04 phase 528-532 in 2007).
Isoflavones belongs to the osajin phytoestrogen, mainly be present in the legume, its main component is genistein, daidzein and Daidezin, have anti-oxidant, regulate the various biological functions such as body immune system and internal system, extremely widely purposes is arranged in herding, culture fishery, and its application prospect is boundless.(Zhang Rui; The characteristic of isoflavones and application study progress thereof, Animal nutrition journal, 11 phase 1884-1890 in 2011.）
Isoflavones can promote growth of animal, and (Wang Kun etc., cysteamine, isoflavones, beta glucan are to eel elver affects on the growth.Fujian agriculture science and technology, 2008,06:75-77; Mi Haifeng etc., Determination of soybean isoflavone in feed and soyasaponins are on the impact of lefteye flounder liver and enteron aisle protein digestibility enzymatic activity and gene expression.Chinese Marine University's journal (natural science edition), 2011,41(12): 40-45), (Huang Guiju etc., isoflavones is to the Primary Study of fish immunity humidification to strengthen immunity of organisms.The south aquatic products, 2005,02:35-40), improve digestive enzyme activity, and (Ye Jidan etc., Daidezin is on the impact of several metabolic enzyme activities in eel growth and liver, the intestinal tissue by improving the active anti-stress ability that strengthens the fish body of antioxidation activity in the body and ACP.Collects The American University's journal (natural science edition), 2008,13(1): 1-6).Simultaneously, isoflavones also has the effect that strengthens immunologic function.Studies show that SPL increment, the transformation of isoflavones energy obvious stimulation mouse, promote mouse delayed allergy effect, improve mouse antibodies cellulation number and serum hemolysin, promote mouse monokaryon-macrophage carbon to clean up effect, the monokaryon of enhancing mouse-peritoneal macrophage phagocytic activity, it is active to improve NK cells in mice.(Han Yanbin etc.; The research that isoflavones affects immune function of mice, Chinese Journal of Health Laboratory Technology, 2010(05): 1046-1048+1058).
Genistein is main isoflavones components in the soybean, therefore, can represent by the content that detects genistein the relevant anti-content that stress bioactie agent of isoflavones in the albumen concentrate feed.Proved and can improve the immune growth of livestock and poultry animal, the enhanced machine body immunity function (thanks to development etc.; The isoflavones genistein is on the impact of laying hen periphery blood T lymphocyte conversion power, and the 8th academic conference of the biochemical branch of animal physiological and national ruminant animal nutrition Physiology and biochemistry be scientific seminar for the third time, and 2004, Changchun; Wang Yang etc., isoflavone tablet are on the impact of the young cock immunologic function of meat, and Chinese agronomy is circulated a notice of, 20 phase 35-37 in 2009; Golden light is bright etc., and isoflavones is on the impact of duck small intestinal mucosa endolymph cell distribution, Anhui Science and Technology College's journal, 2011 years 02 phase 69-72; Yaghoubi SM, Ghorbani GR, Rahmani HR, Nikkhah A. Growth, weaning performance and blood indicators of humoral immunity in Holstein calves fed supplemental flavonoids. J Anim Physiol Anim Nutr (Berl). 2008 Aug; 92 (4): 456-62. Epub 2007 Sep 14.); Genistein also have the physiologically actives such as antibacterial and non-oxidizability (Wang Haitao etc., the research of the bacteriostatic activity of genistein and mechanism thereof, Journal of Nutrition, 2008,30(4): 403-406; Xie Mingjie etc., the bacteriostasis of isoflavones, Soybean Science, 2004,23:101-105).
Bacteriostasis can reduce the pathogenic microorganism stressor; Antioxidation activity can the Scavenger of ROS free radical, regulates by the Radical Metabolism that stress cause disorderly; The raising of immunologic function also strengthens the resistant function to disease.Can improve body immunity so genistein (isoflavones) is a class, have the anti-stress factor of anti-oxidation function.
Be rich in chitin (claiming again chitin, chitin) in shrimp shell, the goldbeater's skin albumen, by the N-deacetylation, degradable is shitosan, and chemical name is Chitosan (1-4)-2-amino-B-D glucose.Can represent by the content that detects Glucosamine the content of the anti-stress factor of shitosan class in the albumen concentrate feed.
Shitosan has growth-promoting effect as feed addictive to animal, and can improve disease-resistant, the immunologic function of aquatic livestock, (Liu Xingguo etc., low molecular chitosan is to the research of Tilapia mossambica hepatic tissue oxidation resistance and liver fat content influence to show good antistress function.Sea fishery, 2004,26(4): 291-294; Shen Jinyu etc., immunopotentiator are on the impact of Eriocheir sinensis immunologic function, Zhejiang Agriculture journal, 2004,16 (1): 25-29; Hu Pinhu, the rare earth chitin application in culture of Chinese mitten crab.Aquaculture, 1999,05:13-14; Li Mingchun etc., the effect of the anti-stress damage of CMC.Practical medical magazine, 2008,25(02): 221; Chen Yun's ripples etc., shitosan is on the impact of allogynogenetic crucian carp and antibacterial ability.Shanghai Aquatic Products Univ. 9CN)'s journal, 2006,15(2): 243-246; Wang Hongquan etc., shitosan is on the impact of Growth of Grass Carps Ctenopharyngodon Idellus and muscle nutrition composition.Agricultural University Of Hunan's journal (natural science edition), 2008,34(5): 576-578).Shitosan and Glucosamine have antibacterial and antioxidation (Han Baoqin etc., CMC, aminoglucose hydrochloride and 2-Acetamido-2-deoxy-D-glucose are on the impact of rat Islet cells growth and amount of insulin secretion, the Chinese biological medical engineering, 2004,02:187-191; Zhang Lanjie etc., the preparation of D-aminoglucose hydrochloride and the research of anti-oxidation characteristics, Food Science, 2007,28(04): 103-105), can reduce and stress and regulate by the radical metabolism that stress cause disorderly.Glucosamine all has obvious facilitation to specific cellular immunity function and humoral immune function and non-specific immune function, can strengthen immune function of mice (Fu Chenwei etc., Glucosamine is on the impact of immune function of mice, Journal of Nutrition, Acta Nutrimenta Sinica, 01 phase 51-53+57 in 2007).Glucosamine and derivative thereof also have radical scavenging activity, reducing power, the protective effect (Yang Yan of the oxidative damage of metal complex ability and the large biological molecule that hydroxy radical is induced such as lipid, deoxyribose and protein; Chitosan oligosaccharide, the anti-oxidation function research of Glucosamine and derivative thereof, Chinese Marine University, biochemistry, 2005, thesis for the doctorate).Glucosamine can improve body immunity, has anti-oxidant, removing Scavenging Oxygen Free Radical, is very promising anti-stress factor.
Antibacterial peptide (Antimicruobial peptides) is the small-molecule peptide bioactivator that a class of organism immune system generation is resisted extraneous pathogenic infection.Antibacterial peptide can improve fish, and (Wang Sixin etc., antibacterial peptide are on the impact of Growth of Grass Carps Ctenopharyngodon Idellus performance, and feed is studied, 2011,04:29-31; Wang Sixin etc., antibacterial peptide are on the impact of carp growth performance, and feed is studied, 2012,05:64-65+73), prawn (Wang Guangjun etc., the antibacterial protein application test in culture of Penaeus vannamei, feed industry, 2005,08:33-34) and river crab (Wang Yijuan etc., antibacterial peptide are on the impact of river crab growth, immunity and oxidation resistance, the Jiangsu agricultural sciences, 2011,02:340-343) and bird (Wang Xiuqing etc.; Antibacterial peptide CecropinB is on the impact of growth and development of chickens and immunologic function, Ningxia Medical University's journal, 01 phase 39-41+46 in 2010; Li Li; Cecropin A baecin preparation and to the research of growth of meat chicken and Immunological Effect, Agricultural University of the Inner Mongol, Animal Nutrition and Feed Science, 2010, thesis for the doctorate; Wang Jing; Antibacterial peptide is on the impact of meat chicken production performance and immunologic function research, Shandong Agricultural University, Animal Nutrition and Feed Science, 2007, Master's thesis) daily gain and feed conversion rate, regulate immunity function, minimizing stress.
CN 102293351A discloses a kind of biological feedstuff for the variegated carp cultivation, and dregs of beans, peanut meal, cotton dregs, the dish dregs of rice are mixed, and the adding Bacillus subtillis stirs and ferments in compound; CN 102524535A discloses the compound oligopeptide additive of a kind of feeding animals and plants, with the animal protein raw materials such as processing fent of the vegetable protein raw material such as dregs of beans, the dish dregs of rice, cotton dregs, corn protein powder or vinasse and fish meal, fish, shrimp, crab or aquatic products, make through access lactic acid bacteria, saccharomycete and bacillus solid state fermentation; CN102669483A discloses a kind of mixed plant albumen fish meal, comprises that following fermenting raw materials makes: the mixture of the dish dregs of rice and cotton dregs, fermentation of bacillus subtilis liquid, Enterococcus faecalis fermentation liquid, fermentation of Aspergillus niger liquid, fermentation by saccharomyces cerevisiae liquid and inorganic salts; CN102334601A discloses a kind of egg feedstuff, utilizes lactic acid bacteria and saccharomyces cerevisiae that guar meal is fermented; CN102178092A discloses a kind of bird feed, at first utilizes composite bacteria liquid fermented bean dregs, cotton dregs, vinasse protein feed to make fermentation protein feedstuff, after fermentation protein feedstuff and corn mixed make; CN102178093A discloses a kind of pig feed, and take dregs of beans, cotton dregs, vinasse protein feed and corn as raw material, access saccharomycete, lactic acid bacteria and bacillus subtilis are fermented and form; CN102715341A discloses the feed of a kind of chicken, pig and ox, take wet slag, dregs of beans and the cotton dregs of producing alcohol as raw material, through the complex microbial community fermentation, comprising: lactobacillus, yeast flora, actinomycete group, thread flora etc.; CN102100302A discloses a kind of animal and fowl fodder, is formed by fermentation of bacillus subtilis grouts and wheat bran; CN102488084A discloses a kind of animal and fowl fodder, take stalk, grouts, bran and corn flour as raw material, add a small amount of ammonium sulfate and urea, transfer pH to 5.5-6.5 with 1% limewash, inoculation Geotrichum, lactic acid bacteria, bacillus licheniformis mixed culture fermentation are made; CN101449739 discloses a kind of micro-ecology fermentation protein feedstuff and preparation method thereof, and dregs of beans and silkworm chrysalis are made with bacillus subtilis, saccharomyces cerevisiae, Bacillus acidi lactici and Rhizopus oligosporus mixed culture solid state fermentation; CN102246884A discloses a kind of laying hen protein feed, utilizes the mixed culture solid state fermentations such as bacillus subtilis and lactobacillus bulgaricus to form.But these prior aries are not all utilized polynary prescription and are adopted secondary fermentation production, and the feed of producing do not relate to have anti-stress effect.
 
Summary of the invention
The present invention adopts multiple endogenous binding protein raw material, and distinctive prescription, suitable proportioning, carries out mixed fungus fermentation by special secondary fermentation, obtains functional height, anti-stress effect is good, economy is strong protein feed product.
Protein feed of the present invention adopts plant source protein raw material 35-90%, terrestrial animal endogenous binding protein raw material 3-20%, marine animal endogenous binding protein raw material 2-60%, and the inoculation composite bacteria carries out high temperature, middle temperature two stage fermentations are made.
The present invention also provides a kind of production method of protein feed of the present invention, comprising: plant source protein raw material, terrestrial animal endogenous binding protein raw material, marine animal endogenous binding protein raw material are mixed in proportion; Inoculation hot fermentation composite bacteria carries out one time fermentation; Inoculation mesophilic digestion composite bacteria carries out secondary fermentation.
The invention still further relates to the purposes of protein feed of the present invention in preparation aquatic products or animal and fowl fodder.
Protein feed of the present invention by microbial fermentation, makes separate sources, the different albumen that forms, and with the microbial source mycoprotein, forms the balance of nutritional labeling, is more conducive to the full nutrition demand of fast growth of animal.
For making things convenient for the selection of protein raw materials of the present invention, improve the carbon-nitrogen ratio of feed of the present invention, the present invention preferably adopts dried porcine saluble as terrestrial animal endogenous binding protein raw material, and fish meal and/or shrimp shell meal are as marine animal endogenous binding protein raw material, and dregs of beans, cotton dregs and the dish dregs of rice are as plant source protein.These raw materials all belong in No. 1773 bulletins " feedstuff catalogue scope " of China in June, 2012 Ministry of Agriculture issue.
Among the present invention, can use simultaneously dregs of beans, cotton dregs and the dish dregs of rice as the plant source protein raw material, their ratios in feedstuff can be dregs of beans 20-80%, cotton dregs 3-40%, dish dregs of rice 3-40%.Dregs of beans also contains isoflavones except having the characteristics such as amino acid balance, good palatability, have the raising animal immunizing power, anti-stress effect.The main active of isoflavones is genistein (genistein), Dai (daidzein) and Glycitein (glycitein).By secondary fermentation, the isoflavones in the dregs of beans is converted into genistein etc., thereby has antibacterial, antioxidation activity, and can improve body immunity, antistress function.
Studies show that too high protein itself is exactly that stressor (Chen Pingjie etc., coerce, feed industry, 2007,28 (22): 18-21) by Crowding Stress and operation in the aquaculture.By secondary fermentation, and the little peptide of the intermediate product of protein degradation has the immune regulating action of the animal body of participation, promotes the anti-stress ability of body.And the little Toplink of antibacterial activity suppresses and kills pathogenic microorganism in the animal body, keeps the healthy of fry, improve survival rate (Sun Kenian, the aquiculture animal anti-stress feed addictive, landlocked aquatic products, 2004,10:45-46).Along with deepening continuously of protein digestibility absorption and metabolism rule research, find the little peptide of part can pass animal intestine wall barrier be absorbed enter the circulatory system (Lin Huangying, little peptide mechanism of absorption progress, herding market, 2005,08:76-78).Compare with amino acid, little peptide is more conducive to promote digesting and assimilating and growing of animal, strengthens resistance against diseases, can improve efficiency of feed utilization.Macro-molecular protein in the puffed soybean is degraded into little molecule small peptide, compare with the protein with same amino acid composition, has the growth of animal of promotion performance, strengthen immunity of organisms, promote biological action (the Chen Pei matches etc. such as absorption of trace elements, the growth of promotion intestinal mucosa 26S Proteasome Structure and Function, the Animal nutrition research of little peptide, cultivation and feed, 2008(4): 95-97).
And be rich in chitin (claiming chitin, chitin) in the shrimp shell, goldbeater's skin albumen, by secondary fermentation, degradable is shitosan and Glucosamine.These metabolic intermediates are with glutathione, antibacterial peptide and the bioactive peptide of microorganism secretion, and the fermentation protein concentrate feed that the present invention is produced not only strengthens the immunity of the animal of raising, and show good antistress function.
Dried porcine saluble is to adopt chitterlings setup action raw material, and through biological enzyme hydrolysis, adhere to and oven dry is made, a kind of animal derived feed protein in fact is rich in the amino acid of short-chain peptide and balance; Strengthen animal particularly production performance and the general level of the health of brood; The amino acid and the free amino acid that exist with the short-chain peptide form have higher biologically active.
As marine animal endogenous binding protein raw material, fish meal and shrimp shell meal can be selected a use, also can use simultaneously.If use simultaneously fish meal and shrimp shell meal, their ratios can be that arbitrary proportion is mixed.
Among the present invention, fermented bacterium preferably select bacillus subtilis ( Bacillus subtillis), bacillus licheniformis ( Baclicus lincheniformis), saccharomyces cerevisiae ( Saccharomyces cerevisiae) and Pediococcus acidilactici ( Pediococcus acidilactici).These four kinds of bacterium all belong in No. 1126 bulletin " feed addictive kind catalogue " of China Ministry of Agriculture in December, 2008 issue.Behind the secondary fermentation by composite bacteria, produce polypeptide, active polysaccharide, amino acid, the mesostate such as vitamin, enzyme and anti-stress nutrition element, and isoflavones and genistein, glutathione, shitosan and Glucosamine and antibacterial peptide etc. are anti-stress active factors.Not only strengthen the immunity of the animal of raising, also show good anti-copingability.
In a preferred embodiment, at fermentation stage, at first inoculate bacillus subtilis ( Bacillus subtillis) and bacillus licheniformis ( Baclicus lincheniformis), under 55-65 ℃ of condition high-temperature aerobic fermentation 6-24 hour, then when the culture medium temperature is reduced to 40-50 ℃, inoculate saccharomyces cerevisiae ( Saccharomyces cerevisiae) and Pediococcus acidilactici ( Pediococcus acidilactici), under 30-50 ℃ of condition, carry out middle temperature amphimicrobian fermentation 24-72 hour.
Bacillus subtilis and bacillus licheniformis are the aerobic bacterias that can produce gemma, can tolerate high temperature, high pressure and soda acid, and vitality is strong.Bacillus subtilis and bacillus licheniformis can produce enzyme and the multiple metabolites such as B family vitamin, VC, protease, amylase and lipase such as vitamin B1, B2, B6, the Nutrition and Metabolism that the degraded of feed is digested and assimilated with animal plays facilitation (Li Yongkai etc., fermented feed containing probiotics research and application present situation, herding and animal doctor, 2009,03:90-93); Withered grass and bacillus licheniformis can secrete multiple antibacterial material simultaneously, studies confirm that at present, bacillus subtilis can produce more than 40 kind of antibacterial material, such as subtilin, subtilosin A, antibacterial protein and the polypeptide (Hu Ruiping etc. such as TasA and sublancin, separation and purification and the evaluation of a bacillus subtilis BSD-2 antibacterial peptide, North China agronomy newspaper, 2011,06:201-206).(the bacillus antibacterial peptide is to the inhibitory effect of Fusariumsp in the shrimp culture environment, guangdong agricultural science, 2011 for Sun Yingfeng etc., 18:95-97) report, the bacillus antibacterial peptide is to the inhibitory effect of Fusariumsp in the shrimp culture environment, thereby improves water body environment, reduces stressor.
Contain the very multiple nutritional components such as rich in protein, B family vitamin, fat, sugar, enzyme in the yeast thalline.A large amount of application study evidences, yeast improve animal immunizing power, improve breeding performonce fo animals and reduce stress etc. the aspect all work.The growth promoting function of yeast may be relevant with its contained abundant free amino acid, nucleotides, beta glucan, mannosan etc. with anti-stress effect.Many studies show that added Yeast debris to having significantly improved animal immunizing power, antibiotic diseases prevention ability and anti-stress ability in the feed (Chen Changfu, immune polysaccharide (yeast cell wall) is to being exempted from the regulating action of hybridized prussian carp immune response.Fresh water fishery, 2004,4:55-57; Chen Guofeng etc., yeast extract is on the impact of allogynogenetic crucian carp and anti-stress ability, feed industry, 2009,18:22-25), (Liu Donghui etc., beta glucan and vitamin C are to the effect of Penaeus monodon growth and premunition because be rich in beta glucan in the Yeast debris.Zhongshan University's journal (natural science edition), 2002,04:59-62; Sahoo PK. etc., The effect of dietary immunomodulation upon Edwardsiella tarda vaccination in healthy and immunocompromised Indian major carp (Labeo rohita). Fish Shellfish Immunol. 2002 Jan; 12 (1): 1-16), (Zhuan Chengji etc., chitin is to the growth of Luo pond crayfish fries, Penaeus monodon and the research of fungi-proofing resistant effect for chitin.Marine University Of Zhanjiang's journal, 1998,18(3): 30-34; Liu is identical, and immune polysaccharide is to the research of Penaeus Vannmei effect.Oceanologia et Limnologia Sinica, 1998,29(3): 113-118) and nucleosides (equality, yeast nucleotides is on the impact of megalobrama amblycephala growth performance, anti-oxidation function and premunition.The Animal nutrition journal, 2012, Vol. (3): 583-590) etc.There is the glutathione of obvious antistress function also generally to produce (Jia Xuejie etc., complex mutation and resistance screening high-yield glutathione yeast strain, Chinese Journal of Pharmaceuticals, 2005,10:604-607 with the saccharomycetes to make fermentation method; Jia Jianping etc., the seed selection of high-yield glutathione bacterial strain, the microbiology circular, 2003,04:24-29).So it is one of main bacteria seed that secondary fermentation of the present invention is selected yeast.
Lactic acid bacteria be use the earliest, probio the most widely, it is present universally acknowledged safe food-grade microorganisms, research finds that its metabolite and living bacterial liquid have very strong fungistatic effect to gram-positive bacteria, Gram-negative bacteria, along with the reduction bacteriostasis of pH value grow gradually, contain higher superoxide dismutase (SOD) in the viable bacteria body and in the metabolite, can strengthen animal immune (Li Xiaohui, the kind of microorganism fodder and Main Function, feed industry, 2002,02:30-32).Lactic acid bacteria antibacterial peptide Nisin is universally acknowledged safe antiseptics for natural food; Class II a class antibacterial peptide take Pediocin as representative has that antimicrobial spectrum is wide, stability height, good dispersion, the advantage such as safe and efficient receive much attention (easy West China etc., the antibiotic potentiality Progress of Research on Mining of the antibiotic potentiality Progress of Research on Mining of bacteriocin lab bacteriocin lab, Food Additives Used in China, 2010,01:73-76).
In the present invention, bacterial classification can be individually according to a conventional method from the bacterial classification that low temperature is preserved the streak inoculation of picking bacterial classification cultivate under appropraite condition to the solid plate culture medium, bacterium colony to be grown is inoculated in fluid nutrient medium and carries out shaken cultivation; Then high temperature compound bacteria bacillus subtilis and bacillus licheniformis carry out respectively the solid fermentation cultivation, and the controlled fermentation condition makes it fully produce enzyme, and culture can use as bacterial classification at once, also can dry preservation, reaches one-year age most.In proportion (0.25-2.5): 1 respectively inoculation carry out one time fermentation, or carry out again inoculation fermentation after culture mixed in the viable count ratio.
Yeast and lactic acid bacteria culturers then are inoculated in respectively in the suitable fluid nutrient medium and ferment, because lactic acid bacteria culture be difficult for to preserve, generally use at once, can be with two kinds of bacterium in viable count ratio (1-5) before carrying out secondary fermentation: 1 secondary inoculation.
After the fermentation, the protein feed of acquisition can directly use.Also the above-mentioned material that ferments can be concentrated, make moisture be reduced to material water ratio 12% or lower, again material be sieved, measures.
Method of the present invention can realize by this area method and conventional equipment commonly used.In a concrete embodiment, method of the present invention can request for utilization number is that 200920108353.8 denominations of invention are implemented for disclosed equipment in the Chinese invention patent of " organic waste biochemical processor ".This complete equipment is made of five parts altogether: hot air circulating system, stirring system, air draft and dust-precipitating system, SOS, automatic lifting feed system, and electric-control system.Whole electric-control system is under programme-control, to the temperature in fermenting, pressure, oxygenating amount, circulated air temperature, temperature of outgoing air etc., by specific technological process, effectively regulate, so that material within the default time, is finished overall process fermentation dried and cooling automatically.
Utilize the biochemical processor of putting down in writing in this patent can finish whole sweat of the present invention, the process parameter control of fermentation can use the computer automation hierarchy of control, fully suitable aerobic and amphimicrobian fermentation condition is provided, guarantees that the protein feed technical target of the product has uniformity, stability and repeatable.
The present invention is in connection with this specific embodiment, to of the present invention anti-stress the polynary prescription of fermentation protein concentrated feed, secondary fermentation and production method detailed process thereof be described further:
(1) raw material disposal: with the metering of weighing of dregs of beans, cotton dregs, the dish dregs of rice and dried porcine saluble and fish meal and/or shrimp shell meal, the organic waste biochemical processor of packing into after mixing in proportion.
(2) once inoculation: hot fermentation composite bacteria bacillus subtilis and bacillus licheniformis carry out respectively solid fermentation to be cultivated, and by the viable count ratio: (0.25-2.5): 1 is mixed into the mixed bacteria of once inoculating.
The mixed bacteria for preparing is mixed with raw material.
Then, add water and regulate material water ratio to 45-60%, water transfer is the growing environment that makes the more suitable microorganism quick active of formation.
The inoculum concentration of bacterial classification can be that those skilled in the art obtain according to conventional knowledge and required attenuation degree mensuration.
Add the 12.5-35g mixed bacteria in per kilogram feed product ratio, so that every gram material contains compound bacteria viable count 1.5-2.0 * 10 7
(3) one time fermentation: start biochemical processor, will treat that fermentation materials stirs; Then start auto-programming, the effective control to the temperature of the temperature of the temperature of fermentation materials, circulated air, air draft and fermentation dried indoor pressure makes material reach 55-65 ℃; Keep constant temperature, fermented 6-24 hour.Circulating fan is constantly sent into fresh air in the processor fermentation vat during this time, and equipment stirs material with certain rotating speed (for example 10-20 rev/min) simultaneously.Make microorganism obtain sufficient oxygen, reach suitable bacillus subtilis and bacillus licheniformis growth, breeding requirement.Simultaneously, the thalline amount reproduction in the processor fermentation vat, and synthetic required various enzymes, the large molecule of degrading is organic, is converted into small molecular protein, polypeptide and metabolism oligosaccharides; Simultaneously material high temperature during the fermentation can be killed the harmful microorganism in the raw material, and falls except some ANFs or change the decomposition that its structure is beneficial to microorganism.
(4) secondary inoculation: after finishing one time fermentation, when temperature of charge is reduced to 40 ℃-50 ℃, inoculation yeast and lactic acid bacteria culturers liquid culture, in viable count ratio (1-5): 1 mixes, the per kilogram material adds the 20-60g mixed bacteria, so that every gram material contains compound bacteria viable count 1.5-2.0 * 10 7This determines that by yeast and lactobacter growth temperature the secondary inoculation applying liquid spawn can make bacterial classification enter fast secondary fermentation simultaneously, also moisture content of material behind the hot fermentation is replenished, and makes it be stabilized in 45-60%.
(5) secondary fermentation: by adjusting the computer automation hierarchy of control of biochemical processor, to the temperature in fermenting, pressure, oxygenating amount, circulated air temperature, temperature of outgoing air etc., by specific technological process, effectively regulate, make material reach 30-50 ℃; Keep constant temperature, fermented 24-72 hour.Make bioactive mesostate such as vitamin are arranged in a large number, glucan, shitosan, genistein, glutathione, Glucosamine, antibacterial peptide etc. can form, thereby make the fermentation protein feedstuff product have very strong anti-stress effect.
(6) processing after: after the fermentation, the fermentation protein feedstuff of acquisition can directly use.Exist with product formation for convenient, preferably that the above-mentioned material that ferments is concentrated, make moisture be reduced to material water ratio 12% or lower, again material is sieved, measures.
Protein feed product of the present invention, it is as shown in the table to obtain after testing its technical indicator:
Table 1 product main nutrient composition
Test item Technical indicator
Crude protein, % 40-60
Crude fibre, % 3-11
Coarse ash, % 6-14
Crude fat, % 1-9
Moisture, %, 5-12
Table 2 product main function material
Test item Technical indicator
Genistein, mg/kg 51-204
Glutathione, mg/kg 34-430
Glucosamine, mg/kg 20-146
Except containing nutritional labeling shown in the upper table and functional materials, BGB of the present invention anti-calcium content 0.2-3.5%, phosphorus content 0.4-1.5% that stress the fermentation protein concentrated feed, salt 0.15-4%, sour molten protein content 4-20%, VBN 15-150mg/100g etc.By special polynary prescription and secondary fermentation, make BGB anti-stress the nutrition of fermentation protein concentrated feed balance, quality are better more; Containing simultaneously the anti-of multiple pricing costliness stress additive; Compound anti-stress effect makes anti-stress effect more obvious, but that cost ratio adds anti-albumen concentrate feed that stress additive is low, more gets consumer reception.
Albumen concentrated feed product that the present invention produces has largely reduced anti-nutrient substance in the vegetable protein such as trypsin inhibitor, urase, hemagglutinin etc., ANFs falls except more than the rate 50%-90%, effectively eliminated protein antigenicity, reduce stressor and anti allergic reaction, be conducive to growing of animal and absorbing of enteron aisle.
Falling except effect (%) of table 3 ANFs
ANFs Fall except rate (%)
Trypsin inhibitor ≥60
The 11S glycinin ≥75
7S β-companion's globulin ≥75
Oligosaccharides ≥70
Phytic acid ≥25
Gossypol ≥60
Glucosinolate ≥60
Of the present invention resisting stress be adopted multicomponent raw material prescription and secondary fermentation by the fermentation protein concentrate feed, be rich in the mesostate that produces in the microorganism secondary fermentation process, such as active polysaccharide, glutathione, antibacterial peptide, enzyme, vitamin, Glucosamine etc., show good anti-stress effect.Genistein, glutathione, shitosan and Glucosamine and antibacterial peptide etc. are anti-stress the active factors total amount to reach 105-780 mg/kg.Thereby make the BGB fermentation protein feedstuff do not need extra add expensive anti-stress additive, not only low price can strengthen the immunity of the animal of raising again, and shows good anti-stress ability.
Resisting that the present invention produces stress be adopted multicomponent raw material prescription and secondary fermentation by the fermentation protein concentrate feed, except producing genistein, glutathione, shitosan and Glucosamine, and antibacterial peptide etc. is anti-stress active factors outside, simultaneously with the digested small-molecule substance that is decomposed into of macromolecular substances complicated in the raw material, poisonous and harmful substance is removed, stressor reduces in the nutritional labeling, increased simultaneously polypeptide, active polysaccharide, amino acid, vitamin, the mesostates such as enzyme and the nutrient that easily is absorbed and used, in aquatic product matched feed, the straight fiery fish meal of energy 3% equivalent substitution, or 4% substitute 3% steam fish meal and 1% powder, has the characteristics of concentrate feed.And can reduce mixed feed cost more than 50 yuan/tons, and do not affect production performance.
 
Prove the effect of feed of the present invention below by experiment.
1 experimental animal: Eriocheir sinensis
1.1 experimental design:
Choose similar pond, 6 choma borders, 10 mu in each pool mouth is put 600 juvenile crabs in a suitable place to breed for every mu; If 2 processed group (control group, aquatic feeds product treatment group), every group of 3 repetitions.The mode of feeding is quantitative bait casting of fixed place and time, free choice feeding.Feed product addition of the present invention is 30% in the test group mixed feed.
1.2 index determining and method:
1.2.1 production performance index: grazing speed, average feed intake, average weight gain, average specification, survival rate, per mu yield, mu benefit;
1.2.2 breeding environment: dissolved oxygen, ammonia nitrogen, nitrite, total nitrogen, total phosphorus;
1.2.3 meat index: amino acid is carried out according to GB/T 5009.124-2003; Volatile materials etc. adopt the GC-14C gas chromatograph to detect.
1.3 result of the test:
The aquatic feeds product that the present invention produces, when adding 30% in river crab compound feed, the river crab grazing speed accelerates 13%, and feed digestibility improves 5%, the river crab weightening finish improves 8%, average specification improves 13.46%, and survival rate improves 16.53%, and water body total nitrogen, ammonia nitrogen reduce about 15%, dissolved oxygen improves 11%, the cultivation per mu yield improves 11.06%, and the mu benefit improves 44.8%, has good economic effect.In addition, dissolved oxygen in water is whole high, and nitrite reduction by 72%, ammonia nitrogen reduction by 17.96%, total nitrogen reduce by 22.06%, and total phosphorus reduces by 9.78%, and breeding environment obviously improves.And, to feed and contain the feed for river crab of aquatic feeds of the present invention, total hydrolysising amino acid content improves 10% in the male crab liver pancreas, total trip amino acid is high by 26.83%, Ester is up to 13.53% in the whole crab, and the fatty acid material has unique flavor characteristic up to 4.05%.
 
2 experimental animals: AA broiler chicken
2.1 experimental design:
1000 AA meat chicks are divided into 5 processed group (conventional corn group, aureomycin processed group, low dosage animal and fowl fodder group, middle dosage animal and fowl fodder group, high dose animal and fowl fodder group) at random, every group of 8 repetitions, each repeats 25 broiler chicken, and male and female is raised together with, free choice feeding.
During the 0-21 age in days, the animal and fowl fodder consumption is that 1-5% adds, and during the 22-42 age in days, the animal and fowl fodder consumption is that 5-15% adds.
2.2 index determining and method:
2.2.1 production performance index: average weight gain, average feed intake, feed conversion rate, the death rate;
2.2.2 breeding environment: H about 2 pm on every Sundays in the duck shed 2S, NH 3, CO 2Concentration;
2.2.3 meat index: pH0 and pH24(0 and 24 represented apart from the time difference of butchering), cooking loss and shearing force, amino acid is carried out according to GB/T 5009.124-2003; Volatile materials etc. adopt the GC-14C gas chromatograph to detect.
2.3 result of the test:
The feed product that the present invention produces when adding 1% in 0-21 age in days AA formula feed of broilers, is compared with the blank group, add 1% group of meat chick average daily gain 28.4g, average daily ingestion amount 45.5g is higher by 14.5% than blank group respectively, 1.1%, feedstuff-meat ratio 1.68 is than blank group low 6.5%; In 22-42 age in days AA meat, in the large chicken mixed feed, add 5% group of meat chick average daily gain 72.1g, average daily ingestion amount 147.6g, higher by 4.3%, 4.7% than blank group respectively, feedstuff-meat ratio 2.06 is than blank group low 1.1%; Product has higher body weight gains and feed intake, preferably feed conversion rate.Feed product of the present invention is on the net in the broiler breeding, ammonia-nitrogen content 5-20mg/kg in the house, dust content, 5-20mg/m 3, breeding environment obviously improves.Feed product of the present invention all 5% adds in the broiler fodder in China, the content 0.24g/100g (dry weight) of 44d meat chick brisket free glutamic acid, and the contrast group is high by 4.34%; The content 0.35g/100g (dry weight) of das Beinfleisch free amino acid, the contrast group is high by 12.9%; Brisket imp content 2027.95 μ g/g(fresh weights), the contrast group is high by 8.93%; Das Beinfleisch imp content 980.6 μ g/g(fresh weights), the contrast group is high by 13.07%; The content of 14 unrighted acids was higher than control group during broiler chicken aliphatic acid formed, and obviously improved the delicate flavour of meat.
 
3 experimental animals: the blue brown laying hen in sea
3.1 experimental design:
600 blue brown laying hens in sea are divided into 5 processed group (control group, 5% animal and fowl fodder group, 10% animal and fowl fodder group, 15% animal and fowl fodder group, 20% animal and fowl fodder group) at random, every group of 6 repetitions, each repeats 20 laying hens.Feeding mode is free choice feeding, drinking-water.
3.2 index determining and method:
3.2.1 production performance index: laying rate, average egg weight, average daily ingestion amount, death rate, feedstuff-egg ratio;
3.2.2 egg matter index: eggshell strength, shell thickness, Hough unit, egg shape index, adopt Egg Quality series instrument (SONOVA company), yolk color adopts Luo Shi colorimetric fan (Switzerland Roche Holding Ag).Volatile materials etc. adopt the GC-14C gas chromatograph to detect.
3.3 result of the test:
The feed product that the present invention produces, in the blue brown laying hen mixed feed in sea, add 10%, when carrying out 60 days feeding experiments, compare with the blank group, can improve yelk bepaint, egg cholesterol content 3.87mg/g, lower by 43.08% than common egg, the linoleic acid of egg yolk, leukotrienes, lecithin content are more than the twice of common egg; Add 5%, egg Hough unit average is 76.44%, and the contrast group improves 4.34%.
 
4 experimental animals: cherry valley duck
4.1 experimental design:
With 4500 Cherry Village Duckss be divided at random 3 processed group (control group, etc. salt replace the corn processed group, etc. nutritive index replace the corn processed group), every group of 5 repetitions, each repeats 300 meat ducks.Feeding mode is free choice feeding.Wherein, testing one group is that the 0-14 age in days adds 5%, 14-32 age in days animal and fowl fodder interpolation 8% with two groups of animal and fowl fodder consumptions of test.
4.2 index determining and method:
4.2.1 production performance index: average weight gain, average feed intake, feedstuff-meat ratio, death rate;
4.2.2 breeding environment: H about 2 pm on every Sundays in the duck shed 2S, NH 3, CO 2Concentration.
4.3 result of the test:
The feed product that the present invention produces adds 5% in the Cherry Village Ducks mixed feed, when carrying out 32 days feeding experiments, daily gain 70.0g, organize than blank high by 2.7%, death rate 0.47%, the contrast group is low by 48.9%, ammonia concentration 7.3-10.7 mg/m in the house 3, than the low 10.8-14.4% of same time point control group ammonia concentration, breeding environment obviously improves.
 
5 experimental animals: Pig Beijing Black
5.1 experimental design:
Pig Beijing Black about 60 51.7kg is divided into 2 processed group (control group, biology corn processed group) at random, every group of 3 repetitions, each repeats 10 pigs.Feeding mode is free choice feeding and drinking-water.Wherein, the test group animal and fowl fodder adds 8%.
5.2 index determining and method:
5.2.1 production performance index: average daily gain, feed intake, feedstuff-meat ratio;
5.2.2 breeding environment: H about 2 pm on every Sundays in the pig house 2S, NH 3, CO 2Concentration.
5.2.3 meat detects: amino acid is carried out according to GB/T 5009.124-2003; Volatile materials etc. adopt the GC-14C gas chromatograph to detect.
5.3 result of the test:
The feed product that the present invention produces, 3% adds in 55-110kg body weight hog grower feed, daily gain 857.14g, the contrast group is high by 4.04%, pig house ammonia level＜20 mg/m 3, hydrogen sulfide content＜10 mg/m 3Breeding environment obviously improves, the content of free glutamic acid is respectively 0.0452g/100g in test group pig leg meat, the lean pork taken under the spinal column of a hog, 0.0786g/100g, the contrast group is high by 13.5%, 13.7% respectively, in volatile materials detects, contain hexyl acetate that control group do not have, octyl acetate etc. in the test group pork, test group pork has the gentle mouthfeel of unique aquatic foods and local flavor.
 
The specific embodiment
Below in conjunction with embodiment, the specific embodiment of the present invention is described in further detail.Following examples are used for explanation the present invention, but are not used for limiting the scope of the invention.
The feed of following examples all both had been suitable for livestock and poultry, also applicable aquatic products.
 
Embodiment 1
Produce as follows fermentation protein feedstuff of the present invention:
1) raw material: dregs of beans, cotton dregs, the dish dregs of rice, shrimp shell meal, dried porcine saluble.
2) once inoculate bacterial classification (hot fermentation composite bacteria): by the viable count ratio of bacillus subtilis, bacillus licheniformis: the solid gemma bacillus agent that 2.5:1 is mixed.
3) one time fermentation: bacterial classification and raw material are mixed, regulate material water ratio to 50-60%, wherein:
Material name Proportioning (%) Batching (kg)
Dregs of beans 20 28
Shrimp shell meal 2 2.8
Cotton dregs 29.5 41.3
The dish dregs of rice 40 56
Dried porcine saluble 5 7
The hot fermentation composite bacteria 3.5 4.9
Subtotal: 100 140
Fermentation time: 6 hours;
Fermentation temperature: 55-65 ℃.
4) secondary inoculation bacterial classification (mesophilic digestion composite bacteria): by the viable count ratio of saccharomyces cerevisiae and Pediococcus acidilactici (liquid): the yeasts lactic acid bacteria agent that 1:1 is mixed.Inoculum concentration 2%.
5) secondary fermentation:
Fermentation time: 72 hours;
Fermentation temperature: 30-45 ℃.
 
Embodiment 2
1) one time fermentation: bacterial classification and raw material are mixed, regulate material water ratio to 50-60%, wherein:
Once inoculate bacterial classification (hot fermentation composite bacteria): by the viable count ratio of bacillus subtilis, bacillus licheniformis: the solid gemma bacillus agent that 2.0:1 is mixed.
 
Material name Proportioning (%) Batching (kg)
Dregs of beans 20 28
Dried porcine saluble 3 4.2
Fish meal 3.5 4.9
Cotton dregs 40 56
The dish dregs of rice 30 42
The hot fermentation composite bacteria 3.5 4.9
Subtotal: 100 140
Fermentation time: 12 hours;
Fermentation temperature: 55-65 ℃.
2) secondary inoculation bacterial classification (mesophilic digestion composite bacteria): by the viable count ratio of saccharomyces cerevisiae and Pediococcus acidilactici (liquid): the yeasts lactic acid bacteria agent that 2:1 is mixed.Inoculum concentration 3%.
3) secondary fermentation:
Fermentation time: 48 hours;
Fermentation temperature: 40-45 ℃.
 
Embodiment 3
1) one time fermentation: bacterial classification and raw material are mixed, regulate material water ratio to 50-55%, wherein:
Once inoculate bacterial classification (hot fermentation composite bacteria): by the viable count ratio of bacillus subtilis, bacillus licheniformis: the solid gemma bacillus agent that 1.5:1 is mixed.
 
Material name Proportioning (%) Batching (kg)
Dregs of beans 55 77
Dried porcine saluble 5 7
Shrimp shell meal 5 7
Cotton dregs 16 22.4
The dish dregs of rice 16.5 23.1
The hot fermentation composite bacteria 2.5 3.5
Subtotal: 100 140
Fermentation time: 18 hours;
Fermentation temperature: 55-65 ℃.
2) secondary inoculation bacterial classification (mesophilic digestion composite bacteria): by the viable count ratio of saccharomyces cerevisiae (solid) and Pediococcus acidilactici (liquid): the yeasts lactic acid bacteria agent that 3:1 is mixed.Inoculum concentration 3%.
3) secondary fermentation:
Fermentation time: 36 hours;
Fermentation temperature: 40-50 ℃.
 
Embodiment 4
1) one time fermentation: bacterial classification and raw material are mixed, regulate material water ratio to 45-50%, wherein:
Once inoculate bacterial classification (hot fermentation composite bacteria): by the viable count ratio of bacillus subtilis, bacillus licheniformis: the solid gemma bacillus agent that 1:1 is mixed.
 
Material name Proportioning (%) Batching (kg)
Dregs of beans 80 112
Dried porcine saluble 7 9.8
Shrimp shell meal 2 2.8
Fish meal 3.5 4.9
Cotton dregs 3 4.2
The dish dregs of rice 3 4.2
The hot fermentation composite bacteria 1.5 2.1
Subtotal: 100 140
Fermentation time: 20 hours;
Fermentation temperature: 55-65 ℃.
2) secondary inoculation bacterial classification (mesophilic digestion composite bacteria): by the viable count ratio of saccharomyces cerevisiae and Pediococcus acidilactici (liquid): the yeasts lactic acid bacteria agent that 4:1 is mixed.Inoculum concentration 4%.
3) secondary fermentation:
Fermentation time: 48 hours;
Fermentation temperature: 35-45 ℃.
 
Embodiment 5
1) one time fermentation: bacterial classification and raw material are mixed, regulate material water ratio to 45-50%, wherein:
Once inoculate bacterial classification (hot fermentation composite bacteria): by the viable count ratio of bacillus subtilis, bacillus licheniformis: the solid gemma bacillus agent that 0.5:1 is mixed.
 
Material name Proportioning (%) Batching (kg)
Dregs of beans 67 93.8
Dried porcine saluble 20 28
Shrimp shell meal 2 2.8
Fish meal 3.5 4.9
Cotton dregs 3 4.2
The dish dregs of rice 3 4.2
The hot fermentation composite bacteria 1.5 2.1
Subtotal: 100 140
Fermentation time: 8 hours;
Fermentation temperature: 55-60 ℃.
2) secondary inoculation bacterial classification (mesophilic digestion composite bacteria): by the viable count ratio of saccharomyces cerevisiae and Pediococcus acidilactici (liquid): the yeasts lactic acid bacteria agent that 4:1 is mixed.Inoculum concentration 5%.
3) secondary fermentation:
Fermentation time: 36 hours;
Fermentation temperature: 35-45 ℃.
 
Embodiment 6
1) one time fermentation: bacterial classification and raw material are mixed, regulate material water ratio to 45-50%, wherein:
Once inoculate bacterial classification (hot fermentation composite bacteria): by the viable count ratio of bacillus subtilis, bacillus licheniformis: the solid gemma bacillus agent that 0.25:1 is mixed.
 
Material name Proportioning (%) Batching (kg)
Dregs of beans 20 28
Dried porcine saluble 5 7
Fish meal 60 84
Cotton dregs 7 9.8
The dish dregs of rice 6.75 9.45
The hot fermentation composite bacteria 1.25 1.75
Subtotal: 100 140
Fermentation time: 6 hours;
Fermentation temperature: 55-60 ℃.
2) secondary inoculation bacterial classification (middle temperature composite fermentation bacterial classification): by the viable count ratio of saccharomyces cerevisiae and Pediococcus acidilactici (liquid): the yeasts lactic acid bacteria agent that 5:1 is mixed.Inoculum concentration 6%.
3) secondary fermentation:
Fermentation time: 24 hours;
Fermentation temperature: 30-40 ℃.
 
Embodiment 7
The protein feed product of embodiment 1-6 gained 59-80 ℃ of lower oven dry, is made moisture be reduced to material water ratio and is lower than 12%, the fermentation protein concentrated feed of gained is carried out technical indicator detect.The examination criteria of all technical is respectively:
(1) conventional feed nutritive index examination criteria is " the Chinese agriculture standard compilation: feed detection method volume " that China Standard Press published distribution on January 1st, 2010.
Crude protein determining method in the GB/T 6432-1994 feed
The mensuration of crude fat in the GB/T 6433-2006 feed
Coarse-fibred assay filtration method in the GB/T 6434-2006 feed
The mensuration of moisture and other volatile matter contents in the GB/T 6435-2006 feed
The mensuration of GB/T 6436-2002 Calcium in Feed
The mensuration AAS of total phosphorus in the GB/T 6437-2002 feed
The mensuration of coarse ash in the GB/T 6438-2007 feed
The mensuration of water soluble chloride in the GB/T 6439-2007 feed
(2) sour molten Protein Detection standard adopts in November, 2008 issue of Standardization Administration of China of General Administration of Quality Supervision, Inspection and Quarantine o of the People's Republic of China, and China Standard Press publishes " the GB/T 22492-2008 soy peptide powder " implemented in January, 2009.
(3) genistein detection method: Su Ju etc., " biotechnology ", 2005,15(4): 54-56
(4) glutathione detection method: Zhu Yaling etc., " Food Science ", 2011,32(06): 251-255
(5) Glucosamine detection method: Zou Qiaogen etc., " Pharmaceutical Biotechnology ", 2008,15 (1): 59-63.
Testing result is as shown in the table:
Table 4 fermentation protein concentrated feed Major Nutrient and functional component detect
Nutritive index and function factor Embodiment 1 Embodiment 2 Embodiment 3 Embodiment 4 Embodiment 5 Embodiment 6
Crude protein, % 40.75 44.45 45.95 48.18 50.45 61.85
Crude fibre, % 9.65 8.97 6.8 4.98 4.46 3.4
Coarse ash, % 7.8 6.57 8.1 7.18 8.61 9.5
Crude fat, % 1.95 1.98 1.8 1.58 2.49 8.0
Moisture, % 10.92 10.65 10.23 9.8 10.5 10.13
Glutathione, mg/kg 34 102 136 204 272 430
Genistein, mg/kg 61 57 109 204 171 51
Glucosamine, mg/kg 39 20 66 60 146 33
As can be seen from the above embodiments, the fermentation protein feedstuff product protein content of producing through the present invention is high, nutritious, and the functional component content such as genistein are high, is applicable to aquatic products, livestock and poultry cultivation and aquatic products, animal and fowl fodder preparation.Simultaneously, the digested small-molecule substance that is decomposed into of macromolecular substances in the fermentation protein concentrated feed that the present invention produces, stressor reduces in the nutritional labeling, having increased simultaneously that genistein, glutathione and Glucosamine etc. are anti-stress active factors, strengthen the immunity of the animal of raising, and showed good anti-stress ability.Carry out multiple batches of processing according to the inventive method, all obtained above basic similarly result.
The % that relates among the present invention if no special instructions outside, all refer to percentage by weight.
The above only is preferred embodiment of the present invention; should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the technology of the present invention principle; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.

Claims (15)

  1. One kind anti-stress fermentation protein feedstuff, it is characterized in that by plant source protein raw material 35-90%, terrestrial animal endogenous binding protein raw material 3-20%, marine animal endogenous binding protein raw material 2-60%, by the inoculation composite bacteria, carry out high temperature, middle temperature two stage fermentations are made, in the described feed, the content of genistein is 51-204 mg/kg, and the content of glutathione is 34-430 mg/kg, and the content of Glucosamine is 20-146 mg/kg.
  2. 2. according to claim 1 anti-stress protein feed wherein uses dried porcine saluble as terrestrial animal endogenous binding protein raw material, and fish meal and/or shrimp shell meal are as marine animal endogenous binding protein raw material, and dregs of beans, cotton dregs and the dish dregs of rice are as plant source protein.
  3. 3. according to claim 2 anti-stress protein feed, wherein as dregs of beans, cotton dregs and the dish dregs of rice of plant source protein raw material, their ratios in feedstuff are dregs of beans 20-80%, cotton dregs 3-40%, dish dregs of rice 3-40%.
  4. 4. according to claim 1 and 2 anti-stress protein feed, wherein at fermentation stage, at first inoculate bacillus subtilis ( Bacillus subtillis) and bacillus licheniformis ( Baclicus lincheniformis), under 55-65 ℃ of condition high-temperature aerobic fermentation 6-24 hour, then when the culture medium temperature is reduced to 40-50 ℃, inoculate saccharomyces cerevisiae ( Saccharomyces cerevisiae) and Pediococcus acidilactici ( Pediococcus acidilactici), under 30-50 ℃ of condition, carry out middle temperature amphimicrobian fermentation 24-72 hour.
  5. 5. according to claim 4 anti-stress protein feed, wherein high-temperature aerobic fermentation is in the stage, bacillus subtilis and bacillus licheniformis are in the viable count ratio: (0.25-2.5): 1 inoculates respectively or is mixed into the once mixed bacteria of inoculation, and the inoculum concentration of fermented bacterium is by this mixed bacteria of per kilogram feed product ratio interpolation 12.5-35g.
  6. 6. according to claim 4 or 5 anti-stress protein feed, wherein in the warm amphimicrobian fermentation stage, yeast and lactic acid bacteria are in viable count ratio (1-5): 1 is mixed into the mixed bacteria of secondary inoculation, and the inoculum concentration of fermented bacterium is pressed this mixed bacteria that the per kilogram material adds 20-60g.
  7. 7. each anti-stress protein feed is wherein concentrated with the material that ferments according to claim 1-6, makes moisture be reduced to material water ratio 12% or lower.
  8. 8. anti-production method that stress fermentation protein feedstuff, it is characterized in that comprising: with plant source protein raw material, terrestrial animal endogenous binding protein raw material, marine animal endogenous binding protein raw material, mix in the ratio of plant source protein raw material 35-90%, terrestrial animal endogenous binding protein raw material 3-20%, marine animal endogenous binding protein raw material 2-60%; Inoculation hot fermentation composite bacteria carries out one time fermentation; Inoculation mesophilic digestion composite bacteria carries out secondary fermentation; Make in the feed of gained, the content of genistein is 51-204 mg/kg, and the content of glutathione is 34-430 mg/kg, and the content of Glucosamine is 20-146 mg/kg.
  9. 9. according to claim 8 anti-stress protein feed production method wherein uses dried porcine saluble as terrestrial animal endogenous binding protein raw material, and fish meal and/or shrimp shell meal are as marine animal endogenous binding protein raw material, and dregs of beans, cotton dregs and the dish dregs of rice are as plant source protein.
  10. 10. according to claim 9 anti-stress protein feed production method, wherein as dregs of beans, cotton dregs and the dish dregs of rice of plant source protein raw material, their ratios in feedstuff are dregs of beans 20-80%, cotton dregs 3-40%, dish dregs of rice 3-40%.
  11. 11. according to claim 8 or 9 anti-stress protein feed production method, wherein at fermentation stage, at first inoculate bacillus subtilis ( Bacillus subtillis) and bacillus licheniformis ( Baclicus lincheniformis), under 55-65 ℃ of condition high-temperature aerobic fermentation 6-24 hour, then when the culture medium temperature is reduced to 40-50 ℃, inoculate saccharomyces cerevisiae ( Saccharomyces cerevisiae) and Pediococcus acidilactici ( Pediococcus acidilactici), under 30-50 ℃ of condition, carry out middle temperature amphimicrobian fermentation 24-72 hour.
  12. 12. anti-stress protein feed production method according to claim 11, wherein high-temperature aerobic fermentation is in the stage, bacillus subtilis and bacillus licheniformis are in the viable count ratio: (0.25-2.5): 1 is mixed into the once mixed bacteria of inoculation, and the inoculum concentration of fermented bacterium is by this mixed bacteria of per kilogram feed product ratio interpolation 12.5-35g.
  13. 13. according to claim 11 or 12 anti-stress protein feed production method, wherein in the warm amphimicrobian fermentation stage, yeast and lactic acid bacteria are in viable count ratio (1-5): 1 is mixed into the mixed bacteria of secondary inoculation, and the inoculum concentration of fermented bacterium is pressed this mixed bacteria that the per kilogram material adds 20-60g.
  14. 14. each anti-stress protein feed production method is wherein concentrated with the material that ferments according to claim 8-13, makes moisture be reduced to material water ratio 12% or lower.
  15. 15. the purposes of the protein feed of claim 1 or 2 in preparation aquatic feeds or animal and fowl fodder.
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