CN112544812A - Mixed-strain fermentation type Chinese herbal medicine additive for laying hens in later period of laying, and preparation method and application thereof - Google Patents
Mixed-strain fermentation type Chinese herbal medicine additive for laying hens in later period of laying, and preparation method and application thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of feed additives, discloses a mixed bacteria fermentation type Chinese herbal medicine feed additive for laying hens in the later period of laying, and provides a mixed bacteria fermentation type Chinese herbal medicine feed additive which can effectively increase the digestion and utilization rate of gastrointestinal tracts to nutrient substances, improve the oxidation resistance level of organisms, delay the degeneration of reproductive systems of the laying hens, prolong the laying period and improve the quality of eggs for the laying hens in the later period of laying. In the process of mixed fermentation of Chinese herbal medicines, aspergillus niger and saccharomyces cerevisiae change own metabolic pathways to adapt to special growth environment, generate new metabolites, generate synergistic effect with the traditional Chinese medicine components, and achieve the effects of improving drug effect, changing drug property, generating new treatment effect and the like. In addition, aromatic substances are generated in the fermentation process, so that the palatability of the feed is improved, and the feeding condition of animals is improved. The invention also provides a preparation method of the mixed bacteria fermentation type Chinese herbal medicine feed additive, which has the advantages of low cost, simple operation process, high content of effective components, richer nutrient substances and better drug effect.
Description
Technical Field
The invention relates to the technical field of feed additives, and particularly relates to a mixed bacteria fermentation type Chinese herbal medicine additive for laying hens in the later period of egg laying, and a preparation method and application thereof.
Background
The feed additive is an important input product in laying hen production, and the antibiotic used as the feed additive in the past decades has been widely used in livestock and poultry breeding production due to the functions of promoting growth and improving feed conversion rate. With the development of feed industry and the wide concern of public on the safety and environmental pollution of animal products, the application of Chinese herbal medicines as substitutes of antibiotic additives in the feeding of laying hens is increasing. The Chinese herbal medicine is used as a green and nuisanceless pure natural feed additive, has rich medicine sources, no toxic or side effect in animals, almost no residue, no drug resistance and dual functions of medicine and nutrition. The Chinese herbal medicine contains a plurality of chemical components, is rich in proteins, amino acids, vitamins, various trace elements, a plurality of organic acids, enzymes, alkaloids, polysaccharides, glycosides, tannins and other active substances which have the effects of nutrition and growth promotion, but a large number of effective components are difficult to release due to the obstruction of plant cell walls. According to the conventional method, the effective components in the Chinese herbal medicines are difficult to be fully dissociated, so that a plurality of effective components are discarded, the dosage is increased, and the economic benefit is reduced. According to the theory of microbial metabolism and fermentation technology, the Chinese herbal medicine is enhanced in efficacy after fermentation, can generate new active ingredients, is obvious in curative effect, has the characteristics of no toxicity, no side effect, no pollution, no drug resistance and the like, and has a very wide application prospect. The number of strains which can be used for producing the microbial fermentation feed is large and tends to the synergistic fermentation of strains, and the synergistic fermentation effect of two or more than two microbes is superior to that of single-bacterium fermentation.
Chinese patent (CN201210056181.0) discloses a traditional Chinese medicine additive for laying hen feed, which comprises: motherwort, prepared rehmannia root, cape jasmine, malaytea scurfpea fruit, dried orange peel, kudzuvine root, medlar, codonopsis pilosula, angelica, bighead atractylodes rhizome, eucommia bark, hawthorn, isatis root, raspberry, ligusticum wallichii, white paeony root, pine needle and hot pepper. The Chinese herbal medicine additive improves the production performance of the laying hens and the quality and the nutritional value of eggs, but the additive cannot effectively improve the gastrointestinal digestion and absorption functions and improve the oxidation resistance of organisms.
Disclosure of Invention
The invention aims to solve the technical problem of overcoming the problems in the prior art and provides a mixed bacteria fermentation type Chinese herbal medicine feed additive for laying hens in the later period of laying.
The second purpose of the invention is to provide a preparation method of the mixed bacteria fermentation type Chinese herbal medicine feed additive for laying hens in the later period of laying.
The third purpose of the invention is to provide the application of the mixed bacteria fermentation type Chinese herbal medicine feed additive for laying hens in the later period of laying.
The purpose of the invention is realized by the following technical scheme:
a mixed bacteria fermentation type Chinese herbal medicine feed additive for laying hens in the later period of egg laying is disclosed, wherein the Chinese herbal medicine comprises the following substances in parts by weight: 5-11 parts of kudzu vine root, 3-9 parts of glossy privet fruit, 3-9 parts of Chinese wolfberry, 2-8 parts of Chinese angelica, 3-9 parts of prepared rehmannia root, 2-8 parts of red paeony root, 5-11 parts of Chinese yam, 3-9 parts of fructus psoraleae, 4-10 parts of combined spicebush root, 2-8 parts of mint, 3-9 parts of cumin, 4-10 parts of schizochytrium limacinum, 4-10 parts of eucommia bark, 5-11 parts of rhodiola rosea, 1-7 parts of cape jasmine fruit and 3-9 parts of longhairy antenoron herb.
Preferably, the Chinese herbal medicine consists of the following substances in parts by weight: 8 parts of kudzuvine root, 6 parts of glossy privet fruit, 6 parts of medlar, 5 parts of Chinese angelica, 6 parts of prepared rehmannia root, 5 parts of red paeony root, 8 parts of Chinese yam, 6 parts of malaytea scurfpea fruit, 7 parts of combined spicebush root, 5 parts of mint, 6 parts of cumin, 7 parts of schizochytrium limacinum, 7 parts of eucommia bark, 8 parts of rhodiola rosea, 4 parts of cape jasmine fruit and 6 parts of longhairy antenoron herb.
Radix Puerariae is root tuber of Pueraria lobata Ohwi of Leguminosae, is sweet and mild in taste and slightly bitter and astringent, and has effects of clearing heat, quenching thirst, promoting fluid production, promoting eruption, removing toxic substance, invigorating yang, relieving diarrhea, activating spleen and relieving hangover. The pueraria isoflavone in the effective bioactive substances of the pueraria plays an important role in regulating the endocrine of female poultry, improving the laying rate and reducing the feed-egg ratio.
The glossy privet fruit is a glossy privet fruit of the family meliaceae, has mild nature, sweet and bitter taste, can strengthen yin, strengthen waist and knee joints, calm five internal organs, nourish spirit, tonify middle-jiao and strengthen tendons and bones, has the efficacies of clearing heat, improving eyesight, supporting healthy energy, and tonifying liver and kidney, and is generally used for treating diseases such as dizziness, tinnitus, dim eyesight, soreness and pain of waist and knees, internal heat, early white beard and hair and the like.
The Chinese wolfberry is a general name of the Chinese wolfberry species such as commercial Chinese wolfberry, plant Ningxia Chinese wolfberry, Chinese wolfberry and the like. The dry mature fruit of Ningxia wolfberry fruit, namely wolfberry fruit, is sweet in nature and neutral, can nourish liver, nourish kidney and moisten lung, and the effective active components of wolfberry polysaccharide and wolfberry pigment have important influence on improving the immune function, antioxidant activity and protecting reproductive function of the organism.
The angelica is a perennial herb belonging to the family of Umbelliferae, and the dried storage root of the angelica is a common Chinese medicinal material in China, has warm taste, enters liver and spleen channels, has the effects of tonifying qi and activating blood, and has wide pharmacological actions of protecting kidney, moistening dryness and lubricating intestines, resisting inflammation and cancer, resisting aging, immunizing and the like.
Radix rehmanniae Preparata (also called dry rehmanniae radix) is prepared from radix rehmanniae of Scrophulariaceae or radix rehmanniae of Huaiqing by processing, steaming and drying in the sun. Sweet in nature and slightly warm in nature, can nourish yin, enrich the blood, replenish vital essence and fill marrow, and has certain pharmacological actions on the aspects of immunity, blood, endocrine, central nervous system, cardiovascular and cerebrovascular diseases, tumor resistance, aging resistance and the like.
The red peony root is the dry root of the Ranunculaceae plant peony or the Sichuan red peony root, is bitter in property and slightly cold, enters liver meridian, can relieve pain and tonify qi, dredge blood vessels, clear heat and cool blood, activate blood and remove stasis, has obvious pharmacological actions of protecting liver, resisting tumor, protecting nerves, protecting heart, resisting thrombus and the like, and the catechin compound in the main chemical active components has strong biological activity of removing free radicals in vivo.
The rhizoma dioscoreae, a dioscorea plant of the dioscoreaceae family, has rich starch, saponin, mucus, choline, glycoprotein and various amino acids in tubers, and has the effects of tonifying deficiency and benefiting vital energy, strengthening spleen and stomach, reducing blood fat and blood sugar, regulating acid-base balance in vivo and improving digestion function of organisms when being used as a medicine.
The fructus psoraleae is bitter, pungent and warm in taste, enters kidney and spleen channels, warms kidney and assists yang, receives qi and relieves asthma, warms spleen and stops diarrhea, and has the activities of resisting inflammation, resisting bacteria and regulating immunity.
The combined spicebush root is the dry root tuber of the combined spicebush root of Lauraceae, pungent and warm in nature, enters lung, spleen, kidney and bladder channels, and is an important analgesic for warming stomach and regulating qi. It can be used for treating cold stagnation, abdominal distention and pain, asthma, bladder deficiency cold, enuresis, pollakisuria, hernia pain, and cold abdominal pain. In addition, the combined spicebush root also has the effects of improving the palatability of the feed, regulating and promoting the digestive function and the like.
The mint is fresh and refreshing, has the effects of dispelling wind and dissipating heat, and can be used for treating symptoms such as cold, fever, laryngalgia, headache, conjunctival congestion, myalgia, pruritus due to rubella on skin, measles without adequate eruption and the like. Menthol contained in mint has various medicinal properties, can relieve abdominal pain and gallbladder problems such as spasm, and also has the effects of antisepsis, sterilization, diuresis, phlegm reduction, stomach strengthening, digestion promotion and the like.
The cumin is rich in various functional nutritional ingredients such as protein, essential fatty acid, polyphenol substances, dietary fiber and the like, has the health-care effects of regulating qi, stimulating appetite, dispelling cold, removing dampness, resisting allergy, sterilizing, inhibiting bacteria, resisting cancer, resisting platelet aggregation, resisting diabetes and the like, and can stimulate appetite and protect the oxidation resistance of an organism.
Schizochytrium limacinum is one of marine microalgae, is rich in essential amino acids, vitamins, mineral elements and various enzymes, and can promote the growth of beneficial microorganisms in healthy intestinal tracts.
Folium Eucommiae is slightly pungent and warm, enters liver and kidney meridians, and is mainly used for treating liver and kidney deficiency, soreness and weakness of waist and knees, flaccidity of tendons and bones, etc. The folium Eucommiae is rich in vitamins, carotene, free amino acids and trace elements, and has antibacterial, antiviral, antifungal, antitumor, antiaging and immunostimulating effects.
The radix Rhodiolae is dry root and rhizome of rhodiola crenulata of Crassulaceae, has sweet, bitter and neutral taste, and enters lung and heart channels, and is mainly used for treating qi deficiency, blood stasis, thoracic obstruction, cardiodynia, apoplexy, hemiplegia, listlessness and asthma. The rhodiola root extract or the effective components thereof have the obvious effects of resisting aging, fatigue, tumor, virus and radiation and the nervous system, the cardiovascular system and the immune system.
Gardenia is dry mature fruit of Gardenia jasminoides Ellis of Rubiaceae, and iridoid glycoside is effective in promoting gallbladder function; the alcohol extract, crocin and crocetin have the effects of promoting bile secretion, promoting pancreas secretion, improving liver and gastrointestinal functions, enhancing the disease resistance of organisms and the like; the gardenia yellow pigment has strong antioxidant, free radical quenching and cancer inhibition activities.
Herba Lysimachiae Christinae belonging to Primulaceae is bitter, sour and slightly cold, and has effects of resisting urinary system calculus, improving cardiovascular system, resisting inflammation and promoting gallbladder.
The kudzu root, the glossy privet fruit, the medlar, the angelica, the prepared rehmannia root and the red paeony root in the Chinese herbal medicine raw materials are qi-tonifying and blood-enriching medicines, have the effects of strengthening body resistance and consolidating the constitution, tonifying qi and consolidating superficial resistance and have estrogen-like effects, and the synergistic effect can stimulate the activity of reproductive organs of poultry, excite the hypothalamus-pituitary-gonadal axis, increase the secretion of blood gonadotropin and promote the development and ovulation of follicles. The Chinese yam, the combined spicebush root, the malaytea scurfpea fruit, the mint and the cumin are Chinese herbal medicines for promoting digestion, removing food stagnation, regulating qi and opening spleen, have the functions of stimulating appetite and improving metabolic rate, can increase calcium storage and pancreatic secretion, and regulate and improve digestive function. The schizochytrium limacinum and eucommia ulmoides are rich in essential amino acids, vitamins and various mineral elements, and have pharmacological effects of resisting bacteria, fungi, viruses and oxidation. In addition, the angelica, the medlar, the cumin, the eucommia leaves, the rhodiola rosea and the like selected by the invention can enhance the oxidation resistance of the organism, inhibit the growth of cancer cells and delay the aging of the organism.
The invention also provides a preparation method of the Chinese herbal medicine feed additive, which takes the mixture of the saccharomyces cerevisiae and the aspergillus niger as a fermentation strain, takes the liquid obtained from the Chinese herbal medicines as a culture medium, and carries out aerobic fermentation, and after the fermentation is finished, the supernatant fluid is the crude product of the feed additive.
Preferably, the preparation method of the Chinese herbal medicine feed additive comprises the following steps:
(1) preparing a bacterial liquid:
preparing a saccharomyces cerevisiae seed solution and an aspergillus niger seed solution;
(2) preparing a Chinese herbal medicine liquid culture medium:
cleaning the Chinese herbal medicines, removing impurities, drying, soaking and sterilizing to obtain the Chinese herbal medicine liquid culture medium.
(3) And (3) fermentation process:
adding pre-cultured Aspergillus niger and yeast seed liquid into the cooled Chinese herbal medicine liquid culture medium, fermenting for 30 days, separating, and sterilizing to obtain the Chinese herbal medicine feed additive.
Preferably, in the step (3), the inoculation amount of the aspergillus niger seed liquid or the yeast seed liquid is 1: 1.2 to 1.5.
More preferably, the above preparation method comprises the steps of:
(1) preparing a bacterial liquid:
inoculating a saccharomyces cerevisiae strain to a malt wort agar culture medium inclined plane, and culturing for 2-4 days in a constant-temperature incubator at 28 ℃ to prepare a saccharomyces cerevisiae seed solution; inoculating aspergillus niger strains to a PDA culture medium inclined plane, culturing in a constant-temperature incubator at 28 ℃, then inoculating into a conical flask filled with a PDB culture medium according to the inoculation amount of 5%, and carrying out shake culture on a constant-temperature shaking table at 28 ℃ for 2-4 days to prepare an aspergillus niger seed solution.
(2) Preparing a Chinese herbal medicine liquid culture medium:
weighing the Chinese herbal medicine components in proportion; firstly, removing impurities such as fibrous roots and silt from kudzuvine roots, rhodiola rosea, Chinese angelica, prepared rehmannia roots, red paeony roots and combined spicebush roots, cleaning schizochytrium limacinum, folium cortex eucommiae, cumin and mint, drying and crushing the Chinese medicinal herbs at 40 ℃, sieving the Chinese medicinal herbs by a 50-mesh sieve to prepare Chinese medicinal powder for later use, finally adding water accounting for 80 percent of the total weight of the Chinese medicinal components into the prepared Chinese medicinal powder, uniformly stirring the mixture, soaking the mixture for 0.5 to 2 hours, adding a proper amount of glucose to adjust the PH value to be about 6.0, carrying out steam sterilization at 100 ℃ for 30min, and cooling the mixture to 30 ℃ to obtain a Chinese.
(3) And (3) fermentation process:
adding pre-cultured Aspergillus niger and yeast seed liquid (according to the inoculum size of 1: 1.5) into the cooled Chinese herbal medicine liquid culture medium; placing the fermentation tank in an environment of about 35 ℃ for aerobic fermentation for 30 days, and ending the fermentation. Centrifuging, drying the concentrated supernatant and the filter residue, mixing, and steam sterilizing to obtain the final product.
The invention also provides application of the Chinese herbal medicine feed additive. The additive is added into the laying hen feed as a feed additive, and the additive can improve the laying rate, improve the quality of eggs and effectively reduce the soft shell egg breaking rate. Meanwhile, the digestive function of the gastrointestinal tract is regulated, the oxidation resistance level of the organism is improved, the aging of the organism is delayed, the egg laying period is prolonged, and the aim of improving the economic benefit of the laying hen breeding industry is fulfilled. In addition, the fermentation product obtained by mixing and fermenting the Chinese herbal medicines is rich in various nutrient substances, the variety of active ingredients of the medicine is increased, and the medicine effect is obviously improved.
Preferably, 0.2 to 1.0 percent of the laying hen feed by weight is added into the daily feed of the laying hen at the later period of egg laying,
preferably, the feed additive is added from 350 days old of laying hens.
Compared with the prior art, the invention has the following beneficial effects:
the invention provides a mixed bacteria fermentation type Chinese herbal medicine feed additive for laying hens in the later period of laying, and provides a fermentation type Chinese herbal medicine feed additive which can effectively increase the digestion utilization rate of gastrointestinal tracts to nutrient substances, improve the oxidation resistance level of organisms, delay the reproductive system degeneration of the laying hens, prolong the laying period and improve the quality of eggs for the laying hens in the later period of laying.
Firstly, the selected herbal medicines have double functions of medicine effect and nutrition, the provided rich nutrient substances can promote the laying hens to absorb and utilize the nutrient substances, improve the intestinal health and the organism immunity function, can clear free radicals in the old laying hens, improve the oxidation resistance state of the organism and delay the aging of the reproductive system, and achieve the aims of improving the egg laying performance and prolonging the production period.
Secondly, the invention can improve the eggshell quality of old laying hens, ensure that the internal quality of eggs is not infected by external bacteria, effectively reduce the loss of eggs in the processes of collection, transportation, storage and the like, and increase the breeding benefit. The Chinese herbal medicines selected by the invention can regulate the pH value of the gastrointestinal tract of the laying hen, increase the action activity of digestive enzyme, inhibit the propagation of harmful bacteria, promote the growth and the planting of beneficial bacteria, improve the absorption and the utilization of nutrient substances, particularly mineral elements such as calcium, phosphorus and the like, and are of great importance for the protection of eggshells. On the other hand, the selected Chinese herbal medicines are rich in vitamins and minerals, can effectively enhance the immune function and the disease-resistant, antibacterial and antioxidant capabilities of the organism, ensure the uterus health of the laying hens and promote the eggshell deposition process.
Thirdly, the aspergillus niger and the saccharomyces cerevisiae are mixed to ferment the Chinese herbal medicine composition, and since the mould can degrade starch and cellulose into monosaccharide or polysaccharide substances which can be more easily utilized by the yeast, the good propagation of the saccharomyces cerevisiae in the fermentation period is promoted. In the process of mixed fermentation of Chinese herbal medicines, aspergillus niger and saccharomyces cerevisiae change own metabolic pathways to adapt to special growth environment, generate new metabolites, generate synergistic effect with the traditional Chinese medicine components, and achieve the effects of improving drug effect, changing drug property, generating new treatment effect and the like. In addition, aromatic substances are generated in the fermentation process, so that the palatability of the feed is improved, and the feeding condition of animals is improved.
The invention also provides a preparation method of the mixed bacteria fermentation type Chinese herbal medicine feed additive, which has the advantages of low cost, simple operation process, high content of effective components, richer nutrient substances and better drug effect.
Detailed Description
The following further describes the embodiments of the present invention. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
All experimental data were subjected to one-way anova and Duncan's multiple comparisons using SPSS 21.0 analysis software.
Example 1
The Chinese herbal medicine feed additive comprises the following Chinese herbal medicine components: every 100 parts (weight parts) of the Chinese herbal medicine composition comprises the following components (in parts by weight): 5 parts of kudzuvine root, 8 parts of glossy privet fruit, 3 parts of medlar, 6 parts of Chinese angelica, 8 parts of prepared rehmannia root, 6 parts of red paeony root, 10 parts of Chinese yam, 9 parts of malaytea scurfpea fruit, 4 parts of combined spicebush root, 2 parts of mint, 3 parts of cumin, 10 parts of schizochytrium, 8 parts of eucommia bark, 5 parts of rhodiola rosea, 5 parts of cape jasmine fruit and 8 parts of longhairy antenoron herb.
The preparation method of the Chinese herbal medicine feed additive comprises the following steps:
(1) preparing a bacterial liquid:
inoculating a saccharomyces cerevisiae strain to a malt wort agar culture medium inclined plane, and culturing for 2-4 days in a constant-temperature incubator at 28 ℃ to prepare a saccharomyces cerevisiae seed solution; inoculating aspergillus niger strains to a PDA culture medium inclined plane, culturing for 2 days in a constant-temperature incubator at 28 ℃, then inoculating the aspergillus niger strains into a conical flask filled with a PDB culture medium according to the inoculation amount of 5%, and performing shake culture for 2-4 days on a constant-temperature shaking table at 28 ℃ to prepare the aspergillus niger seed liquid.
(2) Preparing a Chinese herbal medicine liquid culture medium:
weighing the following Chinese herbal medicine components in parts by weight: 5 parts of kudzuvine root, 8 parts of glossy privet fruit, 3 parts of medlar, 6 parts of Chinese angelica, 8 parts of prepared rehmannia root, 6 parts of red paeony root, 10 parts of Chinese yam, 9 parts of malaytea scurfpea fruit, 4 parts of combined spicebush root, 2 parts of mint, 3 parts of cumin, 10 parts of schizochytrium, 8 parts of eucommia bark, 5 parts of rhodiola rosea, 5 parts of cape jasmine fruit and 8 parts of longhairy antenoron herb.
Firstly, removing impurities such as fibrous roots and silt from kudzuvine roots, rhodiola rosea, Chinese angelica, prepared rehmannia roots, red paeony roots and combined spicebush roots, cleaning schizochytrium limacinum, eucommia ulmoides, cumin and mint, drying and crushing the Chinese medicinal herbs at 40 ℃, sieving the Chinese medicinal herbs by a 50-mesh sieve to prepare Chinese medicinal powder for later use, finally adding water accounting for 80 percent of the total weight of the Chinese medicinal components into the prepared Chinese medicinal powder, uniformly stirring the mixture, soaking the mixture for 0.5 to 2 hours, adding a proper amount of glucose to adjust the pH value to be about 6.0, carrying out steam sterilization at 100 ℃ for 30min, and cooling the mixture to 30 ℃ to obtain a Chinese medicinal.
(3) And (3) fermentation process:
adding pre-cultured Aspergillus niger or yeast seed liquid (according to the inoculum size of 1: 1.5) into the cooled Chinese herbal medicine liquid culture medium; placing the fermentation tank in an environment of about 35 ℃ for aerobic fermentation for 30 days, and ending the fermentation. Centrifuging, drying the concentrated supernatant and the filter residue, mixing well, and steam sterilizing to obtain the Chinese herbal medicine additive.
Example 2
The Chinese herbal medicine feed additive comprises the following Chinese herbal medicine components: every 100 parts (weight parts) of the Chinese herbal medicine composition comprises the following components (in parts by weight): 8 parts of kudzu root, 6 parts of glossy privet fruit, 6 parts of medlar, 5 parts of angelica, 6 parts of prepared rehmannia root, 5 parts of red paeony root, 8 parts of Chinese yam, 6 parts of malaytea scurfpea fruit, 7 parts of combined spicebush root, 5 parts of mint, 6 parts of cumin, 7 parts of schizochytrium, 7 parts of eucommia bark, 8 parts of rhodiola rosea, 4 parts of cape jasmine fruit and 6 parts of longhairy antenoron herb.
The preparation method of the Chinese herbal medicine feed additive comprises the following steps:
(1) preparing a bacterial liquid:
inoculating a saccharomyces cerevisiae strain to a malt wort agar culture medium inclined plane, and culturing for 2-4 days in a constant-temperature incubator at 28 ℃ to prepare a saccharomyces cerevisiae seed solution; inoculating aspergillus niger strains to a PDA culture medium inclined plane, culturing for 2 days in a constant-temperature incubator at 28 ℃, then inoculating the aspergillus niger strains into a conical flask filled with a PDB culture medium according to the inoculation amount of 5%, and performing shake culture for 2-4 days on a constant-temperature shaking table at 28 ℃ to prepare the aspergillus niger seed liquid.
(2) Preparing a Chinese herbal medicine liquid culture medium:
weighing the following Chinese herbal medicine components in parts by weight: 8 parts of kudzu root, 6 parts of glossy privet fruit, 6 parts of medlar, 5 parts of angelica, 6 parts of prepared rehmannia root, 5 parts of red paeony root, 8 parts of Chinese yam, 6 parts of malaytea scurfpea fruit, 7 parts of combined spicebush root, 5 parts of mint, 6 parts of cumin, 7 parts of schizochytrium, 7 parts of eucommia bark, 8 parts of rhodiola rosea, 4 parts of cape jasmine fruit and 6 parts of longhairy antenoron herb.
Firstly, removing impurities such as fibrous roots and silt from kudzuvine roots, rhodiola rosea, Chinese angelica, prepared rehmannia roots, red paeony roots and combined spicebush roots, cleaning schizochytrium limacinum, eucommia ulmoides, cumin and mint, drying and crushing the Chinese medicinal herbs at 40 ℃, sieving the Chinese medicinal herbs by a 50-mesh sieve to prepare Chinese medicinal powder for later use, finally adding water accounting for 80 percent of the total weight of the Chinese medicinal components into the prepared Chinese medicinal powder, uniformly stirring the mixture, soaking the mixture for 0.5 to 2 hours, adding a proper amount of glucose to adjust the pH value to be about 6.0, carrying out steam sterilization at 100 ℃ for 30min, and cooling the mixture to 30 ℃ to obtain a Chinese medicinal.
(3) And (3) fermentation process:
adding pre-cultured Aspergillus niger or yeast seed liquid (according to the inoculum size of 1: 1.2) into the cooled Chinese herbal medicine liquid culture medium; placing the fermentation tank in an environment of about 35 ℃ for aerobic fermentation for 30 days, and ending the fermentation. Centrifuging, drying the concentrated supernatant and the filter residue, mixing well, and steam sterilizing to obtain the Chinese herbal medicine additive.
Example 3
The Chinese herbal medicine feed additive comprises the following Chinese herbal medicine components: every 100 parts (weight parts) of the Chinese herbal medicine composition comprises the following components (in parts by weight): 11 parts of kudzuvine root, 5 parts of glossy privet fruit, 8 parts of medlar, 3 parts of Chinese angelica, 3 parts of prepared rehmannia root, 3 parts of red paeony root, 5 parts of Chinese yam, 8 parts of malaytea scurfpea fruit, 10 parts of combined spicebush root, 8 parts of mint, 9 parts of cumin, 4 parts of schizochytrium, 6 parts of eucommia bark, 11 parts of rhodiola rosea, 2 parts of cape jasmine fruit and 4 parts of longhairy antenoron herb.
The preparation method of the Chinese herbal medicine feed additive comprises the following steps:
(1) preparing a bacterial liquid:
inoculating a saccharomyces cerevisiae strain to a malt wort agar culture medium inclined plane, and culturing for 2-4 days in a constant-temperature incubator at 28 ℃ to prepare a saccharomyces cerevisiae seed solution; inoculating aspergillus niger strains to a PDA culture medium inclined plane, culturing for 2 days in a constant-temperature incubator at 28 ℃, then inoculating the aspergillus niger strains into a conical flask filled with a PDB culture medium according to the inoculation amount of 5%, and performing shake culture for 2-4 days on a constant-temperature shaking table at 28 ℃ to prepare the aspergillus niger seed liquid.
(2) Preparing a Chinese herbal medicine liquid culture medium:
weighing the following Chinese herbal medicine components in parts by weight: 11 parts of kudzuvine root, 5 parts of glossy privet fruit, 8 parts of medlar, 3 parts of Chinese angelica, 3 parts of prepared rehmannia root, 3 parts of red paeony root, 5 parts of Chinese yam, 8 parts of malaytea scurfpea fruit, 10 parts of combined spicebush root, 8 parts of mint, 9 parts of cumin, 4 parts of schizochytrium, 6 parts of eucommia bark, 11 parts of rhodiola rosea, 2 parts of cape jasmine fruit and 4 parts of longhairy antenoron herb.
Firstly, removing impurities such as fibrous roots and silt from kudzuvine roots, rhodiola rosea, Chinese angelica, prepared rehmannia roots, red paeony roots and combined spicebush roots, cleaning schizochytrium limacinum, eucommia ulmoides, cumin and mint, drying and crushing the Chinese medicinal herbs at 40 ℃, sieving the Chinese medicinal herbs by a 50-mesh sieve to prepare Chinese medicinal powder for later use, finally adding water accounting for 80 percent of the total weight of the Chinese medicinal components into the prepared Chinese medicinal powder, uniformly stirring the mixture, soaking the mixture for 0.5 to 2 hours, adding a proper amount of glucose to adjust the pH value to be about 6.0, carrying out steam sterilization at 100 ℃ for 30min, and cooling the mixture to 30 ℃ to obtain a Chinese medicinal.
(3) And (3) fermentation process:
adding pre-cultured Aspergillus niger or yeast seed liquid (according to the inoculum size of 1: 1.5) into the cooled Chinese herbal medicine liquid culture medium; placing the fermentation tank in an environment of about 35 ℃ for aerobic fermentation for 30 days, and ending the fermentation. Centrifuging, drying the concentrated supernatant and the filter residue, mixing well, and steam sterilizing to obtain the Chinese herbal medicine additive.
Comparative example 1
The formulation of the herbal medicine of this comparative example is the same as that of example 1, except that the fermentation method is different from that of example 1.
The fermentation process of this comparative example comprises the following steps:
(1) preparing a bacterial liquid:
and inoculating the saccharomyces cerevisiae strain to the inclined plane of the wort agar culture medium, and culturing for 2-4 days in a constant-temperature incubator at 28 ℃ to prepare saccharomyces cerevisiae seed liquid.
(2) Preparing a Chinese herbal medicine liquid culture medium:
weighing the following Chinese herbal medicine components in parts by weight: 5 parts of kudzuvine root, 8 parts of glossy privet fruit, 3 parts of medlar, 6 parts of Chinese angelica, 8 parts of prepared rehmannia root, 6 parts of red paeony root, 10 parts of Chinese yam, 9 parts of malaytea scurfpea fruit, 4 parts of combined spicebush root, 2 parts of mint, 3 parts of cumin, 10 parts of schizochytrium, 8 parts of eucommia bark, 5 parts of rhodiola rosea, 5 parts of cape jasmine fruit and 8 parts of longhairy antenoron herb.
Firstly, removing impurities such as fibrous roots and silt from kudzuvine roots, rhodiola rosea, Chinese angelica, prepared rehmannia roots, red paeony roots and combined spicebush roots, cleaning schizochytrium limacinum, eucommia ulmoides, cumin and mint, drying and crushing the Chinese medicinal herbs at 40 ℃, sieving the Chinese medicinal herbs by a 50-mesh sieve to prepare Chinese medicinal powder for later use, finally adding water accounting for 80 percent of the total weight of the Chinese medicinal components into the prepared Chinese medicinal powder, uniformly stirring the mixture, soaking the mixture for 0.5 to 2 hours, adding a proper amount of glucose to adjust the PH value to be about 6.0, carrying out steam sterilization at 100 ℃ for 30min, and cooling the mixture to 30 ℃ to obtain a Chinese medicinal.
(3) And (3) fermentation process:
adding a pre-cultured yeast seed solution (according to the inoculation amount of 1: 1.5) into the cooled Chinese herbal medicine liquid culture medium; placing the fermentation tank in an environment of about 35 ℃ for aerobic fermentation for 30 days, and ending the fermentation. Centrifuging, drying the concentrated supernatant and the filter residue, mixing well, and steam sterilizing to obtain the Chinese herbal medicine additive.
Comparative example 2
The formulation of the herbal medicine of this comparative example is the same as that of example 1, except that the fermentation method is different from that of example 1.
The fermentation process of this comparative example comprises the following steps:
(1) preparing a bacterial liquid:
inoculating aspergillus niger strains to a PDA culture medium inclined plane, culturing for 2 days in a constant-temperature incubator at 28 ℃, then inoculating the aspergillus niger strains into a conical flask filled with a PDB culture medium according to the inoculation amount of 5%, and performing shake culture for 2-4 days on a constant-temperature shaking table at 28 ℃ to prepare the aspergillus niger seed liquid.
(2) Preparing a Chinese herbal medicine liquid culture medium:
weighing the following Chinese herbal medicine components in parts by weight: 5 parts of kudzuvine root, 8 parts of glossy privet fruit, 3 parts of medlar, 6 parts of Chinese angelica, 8 parts of prepared rehmannia root, 6 parts of red paeony root, 10 parts of Chinese yam, 9 parts of malaytea scurfpea fruit, 4 parts of combined spicebush root, 2 parts of mint, 3 parts of cumin, 10 parts of schizochytrium, 8 parts of eucommia bark, 5 parts of rhodiola rosea, 5 parts of cape jasmine fruit and 8 parts of longhairy antenoron herb.
Firstly, removing impurities such as fibrous roots and silt from kudzuvine roots, rhodiola rosea, Chinese angelica, prepared rehmannia roots, red paeony roots and combined spicebush roots, cleaning schizochytrium limacinum, eucommia ulmoides, cumin and mint, drying and crushing the Chinese medicinal herbs at 40 ℃, sieving the Chinese medicinal herbs by a 50-mesh sieve to prepare Chinese medicinal powder for later use, finally adding water accounting for 80 percent of the total weight of the Chinese medicinal components into the prepared Chinese medicinal powder, uniformly stirring the mixture, soaking the mixture for 0.5 to 2 hours, adding a proper amount of glucose to adjust the PH value to be about 6.0, carrying out steam sterilization at 100 ℃ for 30min, and cooling the mixture to 30 ℃ to obtain a Chinese medicinal.
(3) And (3) fermentation process:
adding a pre-cultured Aspergillus niger seed solution (according to the inoculum size of 1: 1.5) into the cooled Chinese herbal medicine liquid culture medium; placing the fermentation tank in an environment of about 35 ℃ for aerobic fermentation for 30 days, and ending the fermentation. Centrifuging, drying the concentrated supernatant and the filter residue, mixing well, and steam sterilizing to obtain the Chinese herbal medicine additive.
Examples of the experiments
300 healthy 54-week-old kalimeris indica gray laying hens with similar body weight and an laying rate of 82% are selected and randomly divided into 6 groups, each group is 5 in number, and each group is 10 in number. The group 1 is a control group, and basic ration (without the Chinese herbal medicine additive) is fed; group 2 was trial group i, with 0.2% herbal additive added to the basal ration (example 1); group 3 was trial group ii, with 0.5% herbal additive added to the basal ration (example 2); group 4 was trial group iii with 1.0% herbal additive added to the basal ration (example 3), group five was comparative example 1 (0.2% herbal additive added to basal ration in the same amount as in example 1), group six was comparative example 2 (0.2% herbal additive added to basal ration in the same amount as in example 1), and the trial ration formula is shown in table 1. The test period consisted of 10 weeks, 2 weeks for the pilot period, and 8 weeks for the plus period. The feeding test is carried out in a chicken farm in the city of cattail in Shanxi, and the chickens in the same group are uniformly distributed in a henhouse during the test and are fed and drunk freely.
Table 1 test ration formula
1. Production Performance
And recording the daily egg number, the feeding amount, the egg weight, the number of the stored chicken and the number of broken eggs of each group of laying hens, weighing the residual feed once a week, and counting the egg laying rate, the feed-egg ratio, the average egg weight, the broken egg rate, the death rate and the like of the laying hens.
The results are shown in Table 2, compared with the control group, the egg production rates of the test group I, the test group II and the test group III are obviously increased (P is less than 0.05), and the egg breaking rate and the feed-egg ratio of the test group I, the test group II and the test group III are all obviously lower than those of the control group (P is less than 0.05); the laying rate and average egg weight of the comparative example 1 group and the comparative example 2 group were improved relative to the control group, but were also lower than those of the test I group; the feed-egg ratio and the egg breakage rate of the comparative example 1 group and the comparative example 2 group are reduced, but are higher than those of the test I group, and the feeding effect of the Chinese herbal medicine additive prepared by mixed fermentation is better than that of the Chinese herbal medicine additive prepared by single fermentation.
TABLE 2 influence of mixed fermentation Chinese herbal medicine additive on laying hen production performance
| Item | Control group | Test group I | Test II group | Test group III | Comparative example 1 group | Comparative example 2 group |
| Laying rate (%) | 81.6c | 87.3a | 88.5a | 87.0a | 84.3b | 83.8b |
| Average egg weight (g) | 54.80 | 55.06 | 55.19 | 55.01 | 54.91 | 54.86 |
| Material to egg ratio | 2.42a | 1.56b | 1.28c | 1.33c | 2.22ab | 2.10ab |
| Percentage of broken eggs (%) | 0.13a | 0.06c | 0.07c | 0.06c | 0.10b | 0.11b |
Note: the data in the same row is marked with no letters or the same lower case letters on the shoulder to indicate that the difference is not significant (P is more than 0.05), and the data in the same row is marked with different lower case letters on the shoulder to indicate that the difference is significant (P is less than 0.05). The following table is the same.
2. Egg shell quality and egg internal quality
In the experiment, 20 eggs were randomly picked from each group at 2, 4, 6, and 8 weeks, and the egg shape index (long diameter/short diameter) was calculated by measuring the long diameter and short diameter of the eggs with a vernier caliper. The egg weight, the eggshell thickness, the eggshell strength, the egg white height, the yolk color and the haugh unit were directly read by an egg quality measuring instrument (japanese DET6500) to calculate the eggshell ratio (eggshell ratio ═ eggshell weight/eggshell weight × 100%), and the eggshell density (eggshell density ═ eggshell weight/surface area of egg). The egg-yolk protein was separated by an egg-white-egg-yolk separator, and the egg-yolk-protein ratio (egg-yolk-protein ratio/egg-white mass) was calculated by measuring the mass of each egg-yolk protein.
As shown in table 3, the eggshell thickness, eggshell strength, and eggshell ratio of the test i, test ii, and test iii groups were significantly increased (P <0.05), and the eggshell density was slightly increased, as compared to the control group. Compared with a control group, the harderian unit and the egg white height of the test I group, the test II group and the test III group are obviously increased (P is less than 0.05), and the color of the egg yolk is slightly increased; the eggshell quality and the internal quality of the comparative example 1 group and the comparative example 2 group are improved relative to the control group, but are lower than those of the test I group, which indicates that the Chinese herbal medicine additive prepared by mixed fermentation is better than that prepared by single fermentation in the whole quality of eggs laid by laying hens fed with the Chinese herbal medicine additive.
TABLE 3 influence of mixed fermentation Chinese herbal medicine additive on egg quality
3. Serum antioxidant enzyme activity
In the test of 2 weeks, 4 weeks, 6 weeks and 8 weeks, 5 chickens are randomly selected from each group, blood is collected from fasting wing veins, serum is prepared, and the activity of serum total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-PX), total antioxidant capacity (T-AOC) and the concentration of Malondialdehyde (MDA) are measured by a kit method.
As shown in Table 4, compared with the control group, the T-SOD and GSH-PX activity of the test I group, the test II group and the test III group are all significantly improved (P <0.05), and the MDA concentration is significantly reduced (P < 0.05); compared with the control group, the serum antioxidant enzyme capacity of the group 1 and the group 2 is obviously improved (P is less than 0.05) and is also obviously lower than that of the group I (P is less than 0.05), which shows that compared with the Chinese herbal medicine additive prepared by single-bacterium fermentation, the Chinese herbal medicine additive prepared by mixed-bacterium fermentation has the feeding effect which is more beneficial to improving the organism antioxidant state of the laying hens.
TABLE 4 influence of mixed fermentation Chinese herbal medicine additive on the oxidation resistance of egg-laying hen serum
| Item | Control group | Test group I | Test II group | Test group III | Comparative example 1 group | Comparative example 2 group |
| T-AOC(U/ml) | 10.39 | 10.37 | 10.41 | 10.43 | 10.37 | 10.39 |
| T-SOD(U/ml) | 296.3c | 334.7a | 324.9a | 324.3a | 312.1b | 315.9b |
| GSH-PX(U/ml) | 6725.0c | 7187.3a | 7756.7a | 7650.0a | 6854.3b | 6871.4b |
| MDA(Umol/l) | 5.24a | 4.94c | 4.21c | 4.18c | 5.12b | 5.09b |
4. Digestive tract pH and digestive enzyme Activity
At 2 weeks, 4 weeks, 6 weeks and 8 weeks of the experiment, 5 chickens fed for 2 hours are randomly selected from each group to be killed, chyme of crop, muscular stomach and duodenum is taken, and chyme pH of the crop, muscular stomach and duodenum is directly measured by a pH meter. The activity of the duodenal amylase, the chymotrypsin and the trypsin is determined according to the corresponding kit instructions.
The results of the digestive enzyme activity assay are shown in Table 5. Compared with a control group, the amylase activity and the chymotrypsin activity of the test I group, the test II group and the test III group are all obviously increased (P <0.05), and the trypsin activity of the test I group, the test II group and the test III group is slightly higher than that of the control group. The digestive enzyme activities of the layers of the comparative example 1 group and the comparative example 2 group are improved compared with the control group, but are also obviously lower than that of the test I group (P < 0.05). The results of the digestive tract assay of the laying hens are shown in Table 6. The duodenal PH was significantly reduced (P <0.05) in test I, test ii and test iii groups compared to the control group, and the PH of the layer crop, the muscular stomach and the duodenum was slightly higher in comparative example 1 and comparative example 2 groups than in test I group. Compared with the Chinese herbal medicine additive prepared by single-bacterium fermentation, the Chinese herbal medicine additive prepared by mixed-bacterium fermentation is easier to digest and absorb by the laying hens.
TABLE 5 influence of mixed fermentation Chinese herbal medicine additive on digestive enzyme activity of layer chicken
| Item | Control group | Test group I | Test II group | Test group III | Comparative example 1 group | Comparative example 2 group |
| Amylase (U/mg) | 60.95c | 64.93a | 64.59a | 66.07a | 63.21b | 63.08b |
| Chymotrypsin (U/g) | 1.47c | 1.69a | 1.71a | 1.64a | 1.53b | 1.55b |
| Trypsin (U/g) | 50.78 | 52.14 | 52.27 | 51.06 | 50.94 | 51.05 |
TABLE 6 influence of mixed fermentation Chinese herbal medicine additive on pH of laying hen digestive tract
| Item | Control group | Test group I | Test II group | Test group III | Comparative example 1 group | Comparative example 2 group |
| Crop | 5.66 | 5.62 | 5.59 | 5.61 | 5.61 | 5.62 |
| Muscular stomach | 3.51 | 3.48 | 3.50 | 3.45 | 3.50 | 3.49 |
| Duodenum | 6.08a | 5.73b | 5.92b | 5.62b | 6.02ab | 5.98ab |
5. Apparent metabolic rate of calcium and phosphorus
The test adopts a total manure collection method to determine the apparent metabolic rate of calcium and phosphorus. The feed intake was accurately recorded for 10 chickens per group, and the excreta were collected for 48 hours continuously and the apparent calcium and phosphorus metabolic rates were calculated on the 2 week day of the experiment. The metabolic tests at week 4, week 6 and week 8 were performed as at week 2. And (3) measuring the contents of calcium and phosphorus by flame atomic absorption spectrometry. The nutrient metabolism rate calculation formula is as follows:
a certain nutrient apparent metabolic rate (%) (W1 × a1 to W2 × a2)/(W1 × a 1);
wherein W1 is the daily feed air-dried feed quality (g); w2 is daily excrement air-dried matter mass (g); a1 and A2 are the contents (%) of certain nutrients (calcium and phosphorus) in the diet and excrement, respectively.
As shown in table 7, the calcium and phosphorus metabolic rates were significantly increased in test i, test ii and test iii groups compared to the control group (P < 0.05); the apparent calcium and phosphorus metabolism rates of the group 1 and the group 2 are improved relative to the control group, but are also obviously lower than that of the group I (P <0.05), which indicates that the Chinese herbal medicine additive prepared by mixed fermentation has reduced apparent calcium and phosphorus metabolism rates and higher calcium and phosphorus contents of egg laying of laying hens after being fed relative to the Chinese herbal medicine additive prepared by single fermentation.
TABLE 7 influence of mixed bacteria fermented Chinese herbal medicine additive on apparent calcium and phosphorus metabolic rate of laying hens
While the preferred embodiments of the present invention have been illustrated and described in detail, it should be understood that modifications and variations can be made by persons skilled in the art in light of the above teachings without inventive faculty. Therefore, any technical solutions that can be obtained by a person skilled in the art through logical analysis, reasoning or limited experiments based on the prior art according to the present inventive concept should be within the scope of protection defined by the present claims.
Claims (8)
1. The mixed bacteria fermentation type Chinese herbal medicine feed additive for laying hens in the later period of laying is characterized in that the Chinese herbal medicine is composed of the following substances in parts by weight: 5-11 parts of kudzu vine root, 3-9 parts of glossy privet fruit, 3-9 parts of Chinese wolfberry, 2-8 parts of Chinese angelica, 3-9 parts of prepared rehmannia root, 2-8 parts of red paeony root, 5-11 parts of Chinese yam, 3-9 parts of fructus psoraleae, 4-10 parts of combined spicebush root, 2-8 parts of mint, 3-9 parts of cumin, 4-10 parts of schizochytrium limacinum, 4-10 parts of eucommia bark, 5-11 parts of rhodiola rosea, 1-7 parts of cape jasmine fruit and 3-9 parts of longhairy antenoron herb.
2. The mixed bacteria fermentation type Chinese herbal medicine feed additive for laying hens in the later period of laying eggs according to claim 1, wherein the Chinese herbal medicine is composed of the following substances in parts by weight: 8 parts of kudzuvine root, 6 parts of glossy privet fruit, 6 parts of medlar, 5 parts of Chinese angelica, 6 parts of prepared rehmannia root, 5 parts of red paeony root, 8 parts of Chinese yam, 6 parts of malaytea scurfpea fruit, 7 parts of combined spicebush root, 5 parts of mint, 6 parts of cumin, 7 parts of schizochytrium limacinum, 7 parts of eucommia bark, 8 parts of rhodiola rosea, 4 parts of cape jasmine fruit and 6 parts of longhairy antenoron herb.
3. The method for preparing the mixed fermentation type Chinese herbal medicine feed additive according to any one of claims 1 or 2, characterized in that a mixture of saccharomyces cerevisiae and aspergillus niger is used as a fermentation strain, liquid obtained from Chinese herbal medicines is used as a culture medium, aerobic fermentation is carried out, and after the fermentation is finished, the supernatant is a crude product of the feed additive.
4. The preparation method of the mixed fermentation type Chinese herbal medicine feed additive according to claim 3, which is characterized by comprising the following steps:
(1) preparing a bacterial liquid:
preparing a saccharomyces cerevisiae seed solution and an aspergillus niger seed solution;
(2) preparing a Chinese herbal medicine liquid culture medium:
cleaning the Chinese herbal medicines, removing impurities, drying, soaking and sterilizing to obtain a Chinese herbal medicine liquid culture medium;
(3) and (3) fermentation process:
adding pre-cultured Aspergillus niger and yeast seed liquid into the cooled Chinese herbal medicine liquid culture medium, fermenting for 30 days, separating, and sterilizing to obtain the Chinese herbal medicine feed additive.
5. The method for preparing the mixed fermentation type Chinese herbal medicine feed additive according to claim 4, wherein in the step (3), the inoculation amount of the Aspergillus niger seed solution or the Saccharomyces cerevisiae seed solution is 1: 1.2 to 1.5.
6. The use of the mixed fermentation Chinese herbal medicine feed additive of any one of claims 1 or 2.
7. The use of claim 6, wherein the feed is added to the daily feed of laying hens at the later stage of egg laying in an amount of 0.2 to 1.0% by weight based on the feed amount of the laying hens.
8. The use according to claim 7, wherein the feed additive is added from the 350-day-old laying hens.
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| CN102578423A (en) * | 2012-03-06 | 2012-07-18 | 福建省农业科学院农业生态研究所 | Chinese herbal medicine feed additive for laying hen and preparation method thereof |
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