CN112868888B - Biological fermentation preparation and preparation method thereof - Google Patents
Biological fermentation preparation and preparation method thereof Download PDFInfo
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- CN112868888B CN112868888B CN202110150958.9A CN202110150958A CN112868888B CN 112868888 B CN112868888 B CN 112868888B CN 202110150958 A CN202110150958 A CN 202110150958A CN 112868888 B CN112868888 B CN 112868888B
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/105—Aliphatic or alicyclic compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K40/00—Shaping or working-up of animal feeding-stuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Food Science & Technology (AREA)
- Animal Husbandry (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Physiology (AREA)
- Botany (AREA)
- Mycology (AREA)
- Birds (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Sustainable Development (AREA)
- Fodder In General (AREA)
Abstract
The invention belongs to the technical field of biological fermentation, and discloses a biological fermentation preparation and a preparation method thereof. Inoculating activated and cultured zymophyte into a solid fermentation culture medium consisting of citrus pulp, green plum fruit extract, bran and urea for fermentation culture to obtain a citrus pulp biological fermentation preparation; the zymophyte consists of aspergillus niger, candida tropicalis, lactobacillus plantarum and bacillus subtilis. According to the invention, through the combination of specific zymophytes and the use of citrus residues as main fermentation raw materials, the obtained fermentation product can be applied to broiler feeding to improve the digestibility of broiler nutrients and promote intestinal development, thereby improving the growth performance of broiler and improving the quality and flavor of meat.
Description
Technical Field
The invention belongs to the technical field of biological fermentation, and particularly relates to a biological fermentation preparation and a preparation method thereof.
Background
With the upgrading of consumption, the demands of people on products such as raw juice raw stock, fruit juice, fruit wine processing and the like are gradually increased, and a large amount of pomace waste is generated. Taking the citrus pulp as an example, the citrus pulp generated every year in China is calculated to be more than 500 million tons. The citrus pulp is rich in soluble polysaccharide, vitamins, crude fat, mineral elements and other nutrient components, and some polyphenol and flavonoid functional substances. In addition, the pomace has high water content and crude fiber, is rich in sugar and acidic molecules with strong polarity, and is inconvenient to store and transport. At present, most of the pomace cannot be effectively and comprehensively utilized and can be directly discarded or buried, so that the environment is polluted, and great resource waste is caused. Therefore, how to realize the high-value utilization of the pomace is a problem to be solved urgently at present.
Meanwhile, with the rapid development of animal husbandry, the problem of feed resource shortage becomes a key factor restricting the development of animal husbandry, and the phenomenon of food competition between people and livestock is increasingly severe. Therefore, the adoption of unconventional feed instead of part of traditional feed becomes one of the approaches for solving the grain shortage of the current feed. The pomace, which is a common agricultural product processing waste, contains the nutritional ingredients and functional ingredients, and also contains anti-nutritional substances such as cellulose, lignin, phytic acid and the like, and is directly fed to influence the palatability and the digestibility of the pomace, so that the digestion and the absorption of animals are not facilitated, the growth performance of the animals is further influenced, and the feasibility of directly using the pomace as a feed is seriously restricted. Therefore, the nutritional and functional improvement of the pomace is the key to realize the high-value use of the pomace. The solid state fermentation technology has the advantages of low cost and simple operation, but most of the existing researches and technologies only solve part of links of the feed application of the fermented citrus pulp, and lack of systematicness and corresponding technology integration, so that the fermented fruit pulp lacks of effective technical support in the application process, and the utilization of fruit pulp resources is severely restricted.
Patent CN 108998391A discloses a composite microbial starter, a biological fermentation preparation prepared by using the starter and a preparation method thereof. Taking the combination of aspergillus niger, saccharomycetes and bacillus subtilis as a leavening agent, taking glucose, sucrose, maltose, dextrin or molasses as a carbon source, taking fresh citrus pulp, rice bran, stone powder and magnesium sulfate as solid raw materials, and performing facultative anaerobic fermentation to obtain the magnesium-rich citrus pulp fermented feed. The fattening pig feeding experiment shows that the fermented feed can influence the meat flavor and taste of the fattening pigs; the laying hen feeding experiment shows that the egg quality can be effectively improved. Firstly, the patent can only use specific deposited strains to achieve the established effect, and the realization of the effect depends on mutual promotion and synergistic interaction after aspergillus niger, saccharomycetes and bacillus subtilis are matched with each other according to a specific proportion, and the defect is that the effect is not enough. Secondly, the fermentation technology needs to additionally add a carbon source, so that the fermentation cost is increased, and the effective utilization of the bacteria to the citrus pulp is correspondingly reduced. Thirdly, the fermentation technique does not reveal the promoting effect of the addition of the nitrogen source on the fermentation effect. Fourth, the fermented feed obtained by the patent technology is applied to animal feeding, and results show that the flavor and the taste of meat and eggs can be improved, but the effect that the fermented feed can increase the digestibility of nutrient substances of broiler chickens and promote the development of intestinal tracts so as to improve the growth performance of the broiler chickens is not disclosed.
Patent CN 109907161A discloses a biological fermentation feed and a preparation method thereof. The biological fermentation feed is prepared by fermenting citrus pulp, sweet potato pulp, pomace, bran and cottonseed cake by using a microbial composite strain, wherein the composite strain consists of lactobacillus fermentum, bacillus subtilis, saccharomyces cerevisiae and candida utilis. However, the patent technology only discloses the influence of the corresponding fermentation raw material proportion, strain proportion, inoculation amount, moisture content, fermentation time, fermentation temperature and the like on the contents of crude protein, free amino acid and the like in a fermented feed product, does not disclose the promotion effect of the addition of a nitrogen source on the fermentation effect, and does not perform corresponding animal feeding application tests and application effects.
The technical problem to be solved by the scheme is as follows: the fermentation process and the nutrient components of the existing fermentation preparation are further improved by matching a proper solid fermentation culture medium and zymophyte.
Disclosure of Invention
Aiming at the defects and shortcomings of the prior art, the invention mainly aims to provide a preparation method of a biological fermentation preparation.
The invention also aims to provide the biological fermentation preparation of the citrus pulp, which is prepared by the method.
According to the invention, by optimizing the solid state fermentation process, including strain screening, nitrogen source addition, fermentation substrate proportion, inoculation amount and fermentation time optimization, anti-nutritional factors in the citrus pulp are reduced, and the content of nutritional and functional components is increased. And the feed additive is applied to broiler feeding, can improve the meat quality, flavor and taste, and also has the effects of increasing the digestibility of broiler nutrients, promoting intestinal development and further improving the growth performance of broiler. The implementation of the invention not only can improve the resource utilization degree of the citrus pulp and the economic benefit of fruit processing enterprises, but also has important guiding significance for realizing the high-efficiency resource utilization of the citrus pulp, developing the grain-saving livestock breeding industry and promoting the virtuous cycle economic development of the fruit husbandry industry.
Meanwhile, the invention combines the citrus pulp, the green plum fruit extract and the bran, the green plum fruit extract contains organic acid, flavonoid glycoside, triterpenoid saponin, polyphenol, olefin and other substances, and the green plum fruit extract is matched with the citrus pulp, so that the digestibility of nutrient substances of the broiler chicken can be effectively improved, the intestinal development can be promoted, and the growth performance of the broiler chicken can be further improved.
Meanwhile, through experiments, the fact that the addition of the green plum fruit extract in the solid culture medium can promote the fermentation speed of the fermentation strain combination, can improve the protein conversion rate to a certain extent and reduce the fiber rate can be found.
The purpose of the invention is realized by the following technical scheme:
a preparation method of a biological fermentation preparation comprises the following preparation steps:
inoculating activated and cultured zymophyte into a solid fermentation culture medium consisting of citrus pulp, green plum fruit extract, bran and urea for fermentation culture to obtain a citrus pulp biological fermentation preparation; the zymophyte consists of aspergillus niger, candida tropicalis, lactobacillus plantarum and bacillus subtilis; the mass ratio of the citrus pulp to the green plum fruit extract to the bran in the solid fermentation medium is 75-85: 1-3: 14-22.
In the above method for preparing a biological fermentation preparation, the activating culture step is: and (3) taking unfrozen strains to inoculate in a solid culture medium for culture, then selecting a single colony growing well on the solid culture medium to inoculate in a liquid culture medium for culture, and then taking a bacterial solution for shake culture.
In the preparation method of the biological fermentation preparation, the mass ratio of the citrus pulp, the green plum fruit extract and the bran in the solid fermentation culture medium is 80:2: 18.
In the preparation method of the biological fermentation preparation, the urea in the solid fermentation culture medium is 0.5-2.0% of the total mass of the citrus pulp, the green plum fruit extract and the bran.
In the above method for producing a biological fermentation preparation, the water content of the solid fermentation medium is controlled to be 50 to 60%.
In the preparation method of the biological fermentation preparation, the fermentation culture temperature is 30 ℃, and the fermentation culture time is 3-8 days.
In the preparation method of the biological fermentation preparation, the inoculation ratio of aspergillus niger, candida tropicalis, lactobacillus plantarum and bacillus subtilis in the zymophyte is 1:1:1: 1.
In the preparation method of the biological fermentation preparation, the inoculation amount of the fermentation bacteria inoculated into the solid fermentation culture medium is 10-20%.
A biological fermentation preparation is prepared by the above method.
The biological fermentation preparation of citrus pulp is applied to broiler feeding.
Compared with the prior art, the invention has the beneficial effects that:
(1) the invention adopts the combination fermentation of four strains of aspergillus niger, candida tropicalis, lactobacillus plantarum and bacillus subtilis, and can obviously increase the content of crude protein in a fermentation product and reduce the content of crude fiber.
(2) The fermentation medium mainly comprises the citrus pulp, can fully utilize nutrient components in the citrus pulp, and obviously improves the resource utilization degree of the citrus pulp.
(3) The fermentation raw material orange residue of the invention has rich carbon source but lacks nitrogen source, and in order to make the microorganism fully utilize the nutrient components in the orange residue, a certain amount of urea is added to supplement the nitrogen source, thereby further improving the content of crude protein in the fermented orange residue. Meanwhile, the invention combines the citrus pulp, the green plum fruit extract and the bran, the green plum fruit extract contains organic acid, flavonoid glycoside, triterpenoid saponin, polyphenol, olefin and other substances, and the green plum fruit extract is matched with the citrus pulp, so that the digestibility of nutrient substances of the broiler chicken can be effectively improved, the intestinal development can be promoted, and the growth performance of the broiler chicken can be further improved. Meanwhile, the green plum fruit extract is added into the solid culture medium, so that the fermentation speed of the fermentation strain combination is promoted, the protein conversion rate can be improved to a certain extent, and the fiber rate is reduced.
(4) According to the invention, through the combination of specific zymophytes and the use of citrus residues as main fermentation raw materials, the obtained fermentation product can be applied to broiler feeding to improve the digestibility of broiler nutrients and promote intestinal development, thereby improving the growth performance of broiler and improving the quality and flavor of meat.
Detailed Description
The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.
Example 1
Respectively activating Aspergillus niger, Candida tropicalis, Lactobacillus plantarum and Bacillus subtilis (unfreezing strain 50u l is taken at the edge of a plate culture medium, inoculated on a corresponding lower culture medium by a scribing method and cultured in a culture medium at 37 ℃, the Lactobacillus plantarum and the Bacillus subtilis are cultured for about 12 hours, and the Candida tropicalis and the Aspergillus niger are cultured for about 48 hours at 30 ℃) to obtain activated strains; then performing shake culture on the activated strain (selecting a single colony growing well on a solid culture medium and inoculating the single colony into 5ml of liquid culture medium, placing lactobacillus plantarum and bacillus subtilis in a shaking table at 37 ℃ for 16h, candida tropicalis and aspergillus niger in a shaking table at 30 ℃ for 18h, then respectively inoculating 1ml of bacterial liquid into 100ml of culture medium, respectively placing lactobacillus plantarum and bacillus subtilis in a shaking table at 37 ℃ and a rotating speed of 160r/min for 24h, and placing candida tropicalis and aspergillus niger in a shaking tableCulturing in a shaking table at 30 deg.C and 160r/min for 48h) to obtain bacterial liquid; inoculating 4 kinds of bacteria solution into solid fermentation culture medium composed of Mandarin orange residue, mume fructus extract (GC-MS analysis of ethyl acetate extraction component in mume fructus alcoholic extract, 1.2.1, sample preparation, 3 rd date of 2005, Korean et al, Lin Production chemical engineering communication), testa Tritici and urea at a mass ratio of 1:1:1, and performing fermentation culture, wherein the total inoculation amount of the bacteria solution is 10%, and the number of viable bacteria is 108。
The inoculum size in this context means the percentage of the volume of the inoculum solution relative to the culture medium; the viable bacteria number refers to the number of viable bacteria contained in each ml of bacterial liquid.
The mass percentage of the citrus pulp in the culture medium is 80%, the mass percentage of the green plum fruit extract is 2%, and the mass percentage of the bran is 18%; the adding amount of urea is respectively 0.5%, 1.0%, 1.5% and 2.0% of the total mass of the citrus pulp and the bran, adjusting the water content of the culture medium to be 60%, and performing fermentation culture at 30 ℃ for 4d to obtain the biological fermentation preparation of the citrus pulp.
Crude protein content is shown in Table 1 below
TABLE 1 Effect of different Urea dosages on crude protein production
| The dosage of urea is% | Crude protein content% |
| 0 | 16.95 |
| 0.5 | 18.76 |
| 1 | 19.25 |
| 1.5 | 20.17 |
| 2 | 20.28 |
Compared with the condition without adding urea, the content of crude protein is obviously improved under the condition of adding urea; and with the continuous increase of the urea addition level, the crude protein content of the fermented citrus pulp is increased, which shows that the addition of the urea is helpful for promoting the generation of crude protein in the fermentation process.
Example 2
Respectively activating Aspergillus niger, Candida tropicalis, Lactobacillus plantarum and Bacillus subtilis (unfreezing strain 50u l is taken at the edge of a plate culture medium, inoculated on a corresponding lower culture medium by a scribing method and cultured in a culture medium at 37 ℃, the Lactobacillus plantarum and the Bacillus subtilis are cultured for about 12 hours, and the Candida tropicalis and the Aspergillus niger are cultured for about 48 hours at 30 ℃) to obtain activated strains; then carrying out shake culture on the activated strain (selecting a single colony growing well on a solid culture medium and inoculating the single colony into a 5ml liquid culture medium, placing lactobacillus plantarum and bacillus subtilis in a shaking table at 37 ℃ for 16h, placing candida tropicalis and aspergillus niger in a shaking table at 30 ℃ for 18h, then respectively taking 1ml of bacterial liquid and respectively inoculating the bacterial liquid into 100ml of culture medium, placing lactobacillus plantarum and bacillus subtilis in a shaking table at 37 ℃ and a rotating speed of 160r/min for 24h, and placing candida tropicalis and aspergillus niger in a shaking table at 30 ℃ and a rotating speed of 160r/min for 48h) to obtain bacterial liquid; inoculating 4 kinds of bacteria liquid into a solid fermentation culture medium composed of orange residue, mume fructus extract, bran and urea at a mass ratio of 1:1:1:1, and performing fermentation culture, wherein the total inoculum size of the bacteria liquid is 15%, and the viable bacteria number is 1 × 108The mass percentage of the citrus pulp in the culture medium is 80%, the mass percentage of the green plum fruit extract is 2%, and the mass percentage of the bran is 18%; adding urea 1.5% of the total weight of Mandarin orange residue, mume fructus extract and testa Tritici, adjusting water content of culture medium to 60%, and fermenting and culturing at 30 deg.C for 4d to obtain Mandarin orangeResidue biological fermentation preparation, namely fermentation group 1.
The dried orange residue group contains orange residue 80 wt%, bran 20 wt%, no urea, and no fermentation bacteria, and the orange residue and bran are pulverized and mixed.
The results of the nutrient measurement before and after fermentation in this example are shown in Table 2 below.
TABLE 2 optimal process conditions for fermentation of citrus pulp nutrients
| Nutrient composition | Dried orange pomace (orange pomace and bran) | Fermentation group 1 |
| Moisture Moisture (%) | 9.61 | 55.15 |
| Crude protein CP (%) | 11.67 | 20.17 |
| Crude fat EE (%) | 2.27 | 2.55 |
| Crude fiber CF (%) | 12.83 | 9.12 |
| Total Flavonoids g/100g | 0.245 | 0.36 |
| Hesperidin (%) | 2.07 | 3.57 |
From the results in table 2, it can be seen that the combination of the proper fermentation process and the formula can increase the content of crude protein, total flavonoids and hesperidin and reduce the content of crude fiber.
Examples 3 and 4
The same as example 2, except that the amounts of citrus pulp, green plum fruit extract and bran in example 3 were 75%, 3%, 22%, respectively;
the amounts of citrus pulp, green plum fruit extract and bran in example 4 were 85%, 1%, 14%, respectively.
In example 3, the content of crude protein is 23.57%, the content of total flavone is 0.27%, and the content of hesperidin is 2.87%;
in example 4, the content of crude protein was 18.35%, total flavonoids 0.45%, and hesperidin 4.91%.
Comparative example 1 (Green plum fruit extract deletion experiment)
Referring to example 2, except that the solid fermentation medium consists of citrus pulp, bran and urea, the mass percentage of the citrus pulp in the medium is 80%, and the mass percentage of the bran is 20%; the adding amount of the urea is 1.5 percent of the total mass of the citrus pulp and the bran;
the obtained product is the fermentation group 2.
TABLE 3 Citrus reticulata blanco residue fermentation process nutrient composition under the missing green plum fruit extract process conditions
The test results show that the green plum fruit extract can promote the fermentation of microorganisms and accelerate the fermentation process of citrus pulp.
Comparative example 2 (Candida tropicalis deletion experiment)
Referring to example 2, except that the bacterial species consists of aspergillus niger, lactobacillus plantarum, bacillus subtilis, in the following ratio: aspergillus niger, lactobacillus plantarum and bacillus subtilis in a ratio of 1:1:1, and the number of viable bacteria is 1 × 108 。
The obtained product is the fermentation group 3.
TABLE 4 Citrus reticulata residue fermentation process nutrient composition under Candida tropicalis missing process conditions
Animal experiment 1
Application of citrus pulp biological fermentation preparation obtained by the invention in broiler feeding
1 test design and test daily ration
The test broilers are selected from healthy yellow-feathered broilers with age of 90 days and similar weight, and are divided into 5 treatment groups according to a single-factor random block test, each treatment group is 6 in number, each treatment group is 30 in number, and the treatment groups are adjusted after weighing the broilers one by one, so that the total weight of each treatment group is not obviously different. The 5 treatment groups were: the control group was fed with a basal diet containing no citrus pulp, and the test groups were supplemented with 10% of dry citrus pulp group and 10% of fermented citrus pulp (the citrus pulp biofermentation preparation obtained in example 2), comparative example 1 (green plum extract-deficient group) and comparative example 2 (candida tropicalis-deficient group), respectively, and the diets among the groups were equal in energy and nitrogen. The test is carried out for 4 days before the test is started, the test period is 30 days, and the test can be carried out by freely eating and drinking water. The experimental basic diet is prepared according to NRC (1994 version) broiler chicken nutrition standard, the control group and the dried orange residue group are prepared into granular materials, and the fermented orange residue is directly added into the feed in an initial state after fermentation, namely an undried state. The composition of the test ration is shown in table 5.
Table 5 test diet composition and nutritional level (%)
2 feeding management
The henhouse is thoroughly cleaned, sterilized and disinfected before the test, and is used after air purification. After the broiler chickens are conveyed to a test base, weighing and grouping are carried out, and the drinking water is added with the compound vitamin to relieve transportation stress. The test material control group and the unfermented group are granular materials, the fermented group is a mixture of granules and fermented citrus pulp, the daily feed intake is recorded, and the growth condition and the health condition of the chicken flocks are observed. The feeding management and environmental conditions of each treatment group are basically consistent.
3 sample Collection
The feed is cut off 4 hours in advance one day before the test is finished, 2 test chickens with the weight close to the average weight are randomly selected from each repeating group to be slaughtered, and the carcass character indexes such as bilateral pectoralis muscles, left unilateral leg muscles, abdominal fat and the like are taken to be measured. The left pectoral muscle was taken to determine the quality of the meat.
4 measurement index and method
4.1 growth Performance
At the beginning and end of the trial, all test chickens were weighed in columns and the feed intake (DFI) per column per day was recorded for calculation of the Average Daily Feed Intake (ADFI), Average Daily Gain (ADG) and feed-to-weight ratio (F/G).
DFI (feed-residue)/number of chickens per column;
ADFI is the average total feed consumption per feeding day;
ADG ═ final weight-initial weight/days of feeding;
F/G＝ADFI/ADG。
4.2 apparent digestibility of nutrients
And (3) when the feeding test is finished, selecting 2 test chickens with the weight close to the average weight repeatedly, treating 6 times repeatedly, transferring into a metabolism cage, continuously collecting feces samples for three days at 8 points in the morning every day, and carrying out a metabolism test. And (3) putting the collected excrement sample into a 65 ℃ drying oven for drying treatment, completely drying the excrement sample, putting the excrement sample into air for moisture regain for 1d, weighing the excrement sample, and crushing the excrement sample to determine the content of each nutrient in the excrement sample.
Apparent nutrient digestibility (%) - (the mass of nutrients in feed-the mass of nutrients in feces)/the mass of nutrients in feed × 100%
4.3 intestinal morphology Observation
1cm of intact part of jejunum and middle section of ileum are respectively taken and put into a fixing solution (4% paraformaldehyde solution) for preservation, and the fixing solution is used for observing the morphology of intestinal tissues. After dehydration, embedding, slicing, hematoxylin-eosin staining and the like, the obtained product is observed under a microscope. Each sample was subjected to total enteroscopy and 10 different fields of view were selected for each section to determine the length of the intact villi and the depth of the crypt.
4.4 slaughter Performance
(1) Live body weight: weight after 8h fasting before slaughter.
(2) Carcass weight: exsanguination, and removing the weight of feather, cuticle, and toe shell.
(3) Breast muscle weight: the weight was peeled off the pectoral muscles on both sides behind the carcass.
(4) Leg muscle weight: the left leg behind the carcass was skinned to remove the bony muscle mass.
(5) Abdominal fat weight: abdominal fat and fat surrounding the muscular stomach.
Half bore ratio (%) — half bore weight/living body weight × 100;
the total open weight (%) -, total open weight/living body weight × 100.
4.5 meat quality
(1) Determination of pH
After slaughter, samples of the left pectoral muscle were taken, three different positions were selected and their pH was measured using a pH meter for 45min and 24h, respectively. The instrument employs a pH meter model HI 8424 from Beijing Hanna Instrument science and technology Limited.
(2) Determination of flesh color
Collecting left pectoral muscle sample after slaughtering, selecting three different positions, and measuring brightness (L) for 45min and 24h with color difference meter*) Red (a)*) And yellowness (b), using an automatic colorimeter model CR-410 from the company of maytans, japan.
(3) Determination of drip loss
After slaughtering, a left pectoral muscle sample is taken, the length, width and height of the meat is about 3cm multiplied by 2cm multiplied by 1cm, the meat is weighed after 45min (W1), then the meat is hung on an air blowing bag by an iron hook, the meat is stored at 4 ℃ in a sealed mode, the sample is taken out after 24h, and the meat is continuously weighed after the surface moisture is absorbed by filter paper (W2).
Water drop loss (%) [ (W1-W2)/W1] × 100.
(4) Determination of cooking loss
Weighing the left pectoral muscle sample after 24h refrigeration treatment (W1), inserting a thermometer into the center of the sample, putting the sample into a self-sealing bag, cooking the sample in a water bath kettle at 100 ℃, taking out the sample when the temperature of the muscle reaches 70 ℃, cooling the sample to room temperature, and weighing the sample (W2).
The cooking loss ratio (%) [ (W1-W2)/W1] × 100.
(5) Determination of tenderness
And (3) synchronously performing with the cooking loss, cooling to room temperature, weighing the muscle, taking 5 cylindrical muscle samples of 5cm multiplied by 1.5cm by a sampler according to the trend of the muscle fiber, measuring 10 shear force values by a tenderness meter, and calculating the average value of the shear force values. The instrument used was an Instron model 4411 tenderizer from Instron corporation, Instron, USA.
And (3) test results:
as shown in table 6, the end weight and average daily gain of the broilers of fermentation group 1 and fermentation group 2 were significantly increased compared to the control group and the dried citrus pulp group; compared with the control group, the dried orange residue group and the fermentation group 3, the feed-weight ratio of the broiler chickens in the fermentation group 1 is obviously reduced.
TABLE 6 influence of fermented Citrus pomace on growth Performance of yellow-feathered broilers
As shown in table 7, the apparent digestibility of crude protein and digestion energy was significantly increased in fermentation group 1 (P <0.05) compared to the control group, the dry citrus pomace group, fermentation group 2 and fermentation group 3, while the apparent digestibility of crude protein and digestion energy was significantly higher in fermentation group 2 than in control group, dry citrus pomace group and fermentation group 3(P < 0.05).
TABLE 7 influence of fermented Citrus reticulata Blanco on apparent digestibility of yellow-feathered broilers
As shown in table 8, in jejunum, fermentation 1 and fermentation 2 had ratios of jejunal villus height and villus height to crypt depth (P <0.05) compared to control, while crypt depth was significantly lower for fermentation 1 and fermentation 2 than for dry citrus pomace (P < 0.05). In the ileum, crypt depth was significantly reduced in fermentation 1 and fermentation 2 compared to control (P <0.05), while the ratio of villus height to crypt depth was significantly reduced in fermentation 1 (P < 0.05).
TABLE 8 influence of fermented Citrus residue on intestinal morphology of yellow-feathered broilers
As shown in table 9, different treatments of the daily ration have no significant effect on the slaughter rate, the half-bore rate, the full-bore rate, the leg muscle rate, the breast muscle rate and the abdominal fat rate of the broiler chickens, which indicates that the addition of the citrus pulp in different fermentation forms to the daily ration has no adverse effect on the slaughter performance of the broiler chickens.
TABLE 9 influence of fermented Citrus pomace on slaughter Performance of yellow-feathered broilers
As shown in table 10, the pH value of 45mi muscle after slaughter of broilers and the yellowness value of 24h after slaughter of broilers in fermentation group 1 and fermentation group 2 were significantly increased (P <0.05) and the drip loss was significantly decreased (P <0.05) compared to the control group. Meanwhile, the yellowness values of the fermentation groups 1 and 2 after slaughtering are also obviously higher than those of the dried citrus pulp groups (P is less than 0.05), which shows that the meat quality of the broiler chicken can be improved by adding 10% of citrus pulp of the fermentation groups 1 and 2 into the ration.
TABLE 10 influence of fermented Citrus reticulata Blume on meat quality of yellow-feathered broilers
As shown in table 11, the intramuscular fat and inosinic acid content of the dorking pectoralis muscles of the broilers of the fermentation groups 1 and 2 were significantly increased (P <0.05) compared to the control group, the dried citrus pulp group, and the fermentation group 3, while the inosinic acid content in the pectoralis muscle of the fermentation group 1 was also significantly higher than that of the fermentation group 2(P < 0.05). Further shows that the quality and flavor of the yellow feather broiler muscle can be effectively improved by adding the citrus pulp which contains the green plum fruit extract and is obtained by fermenting the four bacteria into the daily ration.
TABLE 11 influence of fermented Citrus reticulata Blume on conventional nutritional ingredients of yellow-feathered broiler muscles
As shown in Table 12, compared with the control group, the dried citrus pulp group and the fermentation group 3, the fermentation group 1 and the fermentation group 2 significantly increased the ratios of C18:0, C20:3, DHA and PUFA in the breast muscle, and decreased the ratios of C16:1, C gamma 18:1n-9 and MUFA, indicating that the intake of the diet containing the fermented citrus pulp increased the concentration of functional fatty acids in the breast muscle, and further increased the nutritional value thereof
TABLE 12 Effect of fermented Citrus pomace on fatty acid composition in breast muscle of yellow-feathered broilers
In conclusion, the test result shows that the optimal process parameters are obtained by optimizing the citrus pulp fermentation process. Under the conditions that the water content is 60%, the temperature is 30 ℃, the inoculation ratio is 1:1:1:1, and the addition amount of urea is 1.5%, the ratio of fermentation substrate orange residue, green plum fruit extract and bran is 80:2:18, the inoculation amount is 15%, the fermentation time is 4d, and the fermentation process of the citrus pulp is optimal. The fermented citrus pulp with the concentration of 10% is added into the daily ration of the broiler chicken, so that the digestibility of nutrient substances of the broiler chicken is increased, the intestinal development is promoted, the growth performance of the broiler chicken is improved, and the quality and the flavor of the broiler chicken are improved.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (7)
1. A preparation method of a biological fermentation preparation is characterized by comprising the following steps: the preparation method comprises the following preparation steps:
inoculating activated and cultured zymophyte into a solid fermentation culture medium consisting of citrus pulp, green plum fruit extract, bran and urea for fermentation culture to obtain a citrus pulp biological fermentation preparation; the zymophyte consists of aspergillus niger, candida tropicalis, lactobacillus plantarum and bacillus subtilis; the mass ratio of the citrus pulp to the green plum fruit extract to the bran in the solid fermentation medium is 75-85: 1-3: 14-22;
the urea in the solid fermentation culture medium is equivalent to 0.5-2.0% of the total mass of the citrus pulp, the green plum fruit extract and the bran;
the inoculation ratio of aspergillus niger, candida tropicalis, lactobacillus plantarum and bacillus subtilis in the zymocyte is 1:1:1: 1;
the mume fructus extract contains organic acids, flavone glycoside, triterpene saponin, polyphenol and olefin.
2. The method of claim 1, wherein the step of preparing the biofermentation preparation comprises: the activation culture comprises the following steps: and (3) taking unfrozen strains to inoculate in a solid culture medium for culture, then selecting a single colony growing well on the solid culture medium to inoculate in a liquid culture medium for culture, and then taking a bacterial solution for shake culture.
3. The method of claim 1, wherein the step of preparing the biofermentation preparation comprises: the mass ratio of the citrus pulp to the green plum fruit extract to the bran in the solid fermentation medium is 80:2: 18.
4. The method of claim 1, wherein the step of preparing the biofermentation preparation comprises: the water content of the solid fermentation medium is controlled to be 50-60%.
5. The method of claim 1, wherein the step of preparing the biofermentation preparation comprises: the temperature of the fermentation culture is 30 ℃, and the time of the fermentation culture is 3-8 d.
6. The method of claim 1, wherein the step of preparing the biofermentation preparation comprises: the inoculation amount of the fermentation bacteria inoculated into the solid fermentation culture medium is 10-20%.
7. A biological fermentation preparation, which is characterized in that: prepared by the method of any one of claims 1 to 6.
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| CN113875887B (en) * | 2021-09-16 | 2023-05-23 | 广东省农业科学院动物科学研究所 | Citrus residue-based fermentation preparation and preparation method and application thereof |
| CN114504062B (en) * | 2022-03-28 | 2023-05-23 | 广西壮族自治区畜牧研究所 | Feed for replacing daily ration of spicy chickens and application method of feed |
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