CN112472730A - Chinese mugwort herb micro-ecological fermentation preparation and preparation method and application thereof - Google Patents
Chinese mugwort herb micro-ecological fermentation preparation and preparation method and application thereof Download PDFInfo
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Abstract
A Chinese medicinal microecological fermented preparation of wormwood, a preparation method and an application thereof relate to the field of livestock and poultry breeding, and the preparation comprises lactobacillus rhamnosus YL198 bacterial suspension and wormwood, wherein the weight ratio of the lactobacillus rhamnosus YL198 bacterial suspension to the wormwood is 1L:100 kg-1L: 500 kg; the lactobacillus rhamnosus YL198 is preserved in China center for type culture Collection in 11 months and 27 days of 2020, and the preservation number is CCTCC NO: m2020781. According to the invention, the traditional Chinese medicine wormwood is fermented by the lactobacillus rhamnosus YL198, and the magnetic field generated by the terahertz ore is used for assisting probiotic fermentation and conversion of bioactive components of the wormwood, so that the pH value of the obtained traditional Chinese medicine microecological preparation is 3.0-3.5, the product has the functions of enhancing organism immunity, adjusting intestinal flora, improving intestines and stomach and the like, is green, environment-friendly, safe, effective and stable, and can be used as a feed additive for replacing or reducing antibiotics.
Description
Technical Field
The invention relates to the technical field of livestock and poultry breeding, and particularly relates to a wormwood traditional Chinese medicine micro-ecological fermentation preparation as well as a preparation method and application thereof.
Background
Many food safety issues are due to the detection of antibiotic residues in the livestock and poultry products, which is caused by the fact that the breeders do not comply with national standards or do not strictly comply with the drug holiday in the breeding process. The overuse of antibiotics is always a potential safety hazard in the breeding industry of China, not only is the residue of antibiotics easily caused, but also the long-term use of the antibiotics can cause endogenous infection and superinfection of animals, and the production performance and the service life of the animals are reduced. Wherein, endogenous infection means that antibiotics can cause the dysbacteriosis of microbial flora, so that the beneficial microbial flora in an animal is reduced, and the original drug-resistant pathogenic bacteria in the intestinal tract dominate to cause animal diseases.
All preparations capable of promoting the growth and the reproduction of normal microbiota and inhibiting the growth and the reproduction of pathogenic bacteria are called 'microecologics'. It is prepared by using probiotics which are beneficial to and harmless to a host or growth promoting substances of the probiotics by applying the microecology principle through the processes of culturing, fermenting, drying, processing and the like. The microecological preparation has the characteristics of no toxic or side effect, no drug resistance, no residue, low cost, obvious effect, no environmental pollution and the like, and is gradually accepted by the vast breeding world.
The feed additive enters the micro-ecological era from the antibiotic era, and the research and the application of the probiotic series products have very wide prospect and good economic benefit. The use of current microecologics in the livestock breeding industry presents several problems: the viable bacteria preparation is easily affected by intestinal bile acid, hydrochloric acid and the like, and the play of the function is greatly reduced, so that the genetic engineering technology is dedicated to screening and breeding of beneficial strains, and finally, the strains with better tolerance are selected for modification and development; in addition, the viable bacteria preparation is easy to lose activity in the processes of processing, transportation and storage, so the development of the microcapsule process has better effect on the growth and drying of the probiotics, and the probiotics with high number of stable and efficient viable bacteria can be produced according to the characteristics of the strains; the quality of the product is good and uneven, and the collocation, content and safety of the microecological preparation are uncertain. Therefore, the probiotic preparation should be scientifically prepared and the effectiveness of the probiotic preparation should be exerted to the maximum extent.
Disclosure of Invention
The invention aims to provide a wormwood traditional Chinese medicine micro-ecological fermentation preparation, and a preparation method and application thereof, so as to solve the problems of the existing micro-ecological preparation in the livestock and poultry breeding industry.
The technical scheme adopted by the invention for solving the technical problem is as follows:
the Chinese herbal microecological fermented preparation comprises a Lactobacillus rhamnosus YL198 bacterial suspension and wormwood, wherein the dosage ratio of the Lactobacillus rhamnosus YL198 bacterial suspension to the wormwood is as follows: the weight of the powder is 1L:100 kg-1L: 500 kg;
the Lactobacillus rhamnosus (Lactobacillus rhamnosus) YL198 is preserved in China center for type culture Collection at 11 month and 27 month in 2020, and the preservation numbers are as follows: CCTCC NO: m2020781.
In a preferred embodiment, the ratio of the Lactobacillus rhamnosus (Lactobacillus rhamnosus) YL198 bacterial suspension to the wormwood is as follows: the volume and weight are 1L to 300 kg.
The preparation method of the wormwood traditional Chinese medicine micro-ecological fermentation preparation comprises the following steps:
inoculating lactobacillus rhamnosus YL198 to a liquid MRS culture medium, performing static culture at 37-45 ℃ for 16-25 h, centrifuging at 4-10 ℃ and 4000-8000 rpm for 5-10 min, and collecting thallus precipitates; suspending with aqueous solution of fermentation and sterilization culture medium, adjusting viable count to 1.0 × 109～1.0×1010CFU/ml to obtain YL198 bacterial suspension;
naturally drying the overground part of fresh wormwood, soaking for 2 times by using 0.5-1% of saline solution, cleaning with clear water, placing terahertz ore at the bottom of a fermentation tank, placing the wormwood, covering the wormwood with the terahertz ore, wherein the total weight of the terahertz ore is 1-2% of the total weight of the wormwood, adding YL198 bacterial suspension, covering a plastic film, sealing, fermenting for 10-20 days at constant temperature of 30-35 ℃, performing solid-liquid separation after fermentation is finished, fermenting the wormwood again under the same condition, combining fermentation liquor obtained in two times, filtering, and packaging to obtain the wormwood traditional Chinese medicine micro-ecological fermentation preparation, wherein the pH value is 3.0-3.5.
As a preferred embodiment, the preparation method of the wormwood traditional Chinese medicine micro-ecological fermentation preparation comprises the following steps:
inoculating Lactobacillus rhamnosus YL198 to liquid MRS culture medium, standing at 37 deg.C for 16 hr, centrifuging at 4 deg.C and 6000rpm for 8min, and collecting thallus precipitate; by fermentationSuspending the aqueous solution of the sterilized culture medium, and adjusting the number of viable bacteria to 1.0 × 109CFU/ml to obtain YL198 bacterial suspension;
naturally drying the overground part of fresh wormwood, soaking for 2 times by using 0.75% of edible salt water, cleaning by using clear water, putting terahertz ore at the bottom of a fermentation tank, putting the wormwood, covering the wormwood with the terahertz ore, adding YL198 bacterial suspension, wherein the total weight of the terahertz ore is 1.5% of the total weight of the wormwood, adding the YL198 bacterial suspension and the wormwood according to the volume ratio of 1L to 300kg, covering a plastic film, sealing, carrying out constant temperature of 30 ℃, fermenting for 10 days, carrying out solid-liquid separation after the fermentation is finished, carrying out re-fermentation on the wormwood once under the same condition, combining two fermentation liquids, filtering and packaging to obtain the wormwood traditional Chinese medicine micro-ecological fermentation preparation, wherein the pH value is 3.0-3.5.
In a preferred embodiment, the aqueous solution of the fermentative sterilization medium is prepared by the following method: adding 30g of corn steep liquor powder and 30g of molasses into each liter of liquid culture medium, and autoclaving at 121 deg.C for 15min to obtain the final product.
In a preferred embodiment, the mugwort Chinese medicinal micro-ecological fermentation preparation is in the form of water aqua, powder or tablet.
The Chinese medicinal herb micro-ecological fermentation preparation of the invention is applied as a feed additive.
The invention has the beneficial effects that:
the Lactobacillus rhamnosus (Lactobacillus rhamnosus) YL198 adopted by the invention is preserved in China center for type culture Collection in 11 months and 27 days in 2020, and the preservation numbers are as follows: CCTCC M2020781.
Aiming at the technical requirement of 'nonreactive breeding' of livestock and poultry, the traditional Chinese medicine wormwood is fermented by Lactobacillus rhamnosus YL198, and the magnetic field generated by terahertz ore is used for assisting probiotic fermentation and assisting fermentation conversion of bioactive components of the wormwood to prepare the wormwood traditional Chinese medicine microecological fermentation preparation. The prepared wormwood traditional Chinese medicine micro-ecological fermentation preparation has the functions of enhancing organism immunity, regulating intestinal flora, improving stomach and intestine and the like, and also has the functions of improving the growth performance of livestock and poultry, improving the biochemical index of serum, improving the organism immunity, reducing the expression of serum inflammatory factors, maintaining the micro-ecological balance of the intestinal tract, promoting metabolism, improving meat quality and the like.
The Chinese mugwort herb micro-ecological fermentation preparation prepared by the invention is green, environment-friendly, safe, effective and stable, and can be used as a feed additive for replacing or reducing antibiotics.
The traditional Chinese medicine microbial fermentation preparation provided by the invention can effectively solve the problems of drug resistance generated by antibiotics, residue in animals and the like, thereby providing a more green, safe and environment-friendly choice for feed additives.
Detailed Description
The Chinese herbal microecological fermented preparation mainly comprises Lactobacillus rhamnosus YL198 bacterial suspension and folium artemisiae argyi, and the dosage ratio of the Lactobacillus rhamnosus YL198 bacterial suspension to the folium artemisiae argyi is as follows: the weight of the powder is 1L:100 kg-1L: 500 kg; preferably, the dosage ratio of the Lactobacillus rhamnosus (Lactobacillus rhamnosus) YL198 bacterial suspension to the folium artemisiae argyi is as follows: the volume and weight are 1L to 300 kg.
Wherein, the Lactobacillus rhamnosus (Lactobacillus rhamnosus) YL198 is preserved in China center for type culture Collection at 11/27 of 2020 with the addresses as follows: eight-path Wuhan university school (Wuhan university collection center) 299 in Wuchang area, Wuhan city, Hubei province has the collection number: CCTCC NO: m2020781.
The Chinese medicinal herb microecological fermentation preparation of wormwood of the invention is prepared by fermenting Chinese medicinal herbs serving as pharmacodynamic raw materials through Lactobacillus rhamnosus (Lactobacillus rhamnosus) YL198, and specifically comprises the following steps:
inoculating lactobacillus rhamnosus YL198 to a liquid MRS culture medium, performing static culture at 37-45 ℃ for 16-25 h (preferably, 16h at 37 ℃), centrifuging at 4-10 ℃ and 4000-8000 rpm for 5-10 min (preferably, centrifuging at 4 ℃ and 6000rpm for 8min), and collecting thallus precipitates; suspending with aqueous solution of fermentation and sterilization culture medium, adjusting viable count to 1.0 × 109～1.0×1010CFU/ml to obtain YL198 bacterial suspension;
naturally drying the overground part of fresh wormwood, soaking for 2 times by using 0.5-1% (preferably 0.75%) of salt solution, cleaning with clear water, placing terahertz ore at the bottom of a fermentation tank, placing wormwood, covering the terahertz ore on the wormwood, wherein the total weight of the terahertz ore is 1-2% (preferably 1.5%) of the total weight of the wormwood, adding YL198 bacterial suspension, compacting, covering a plastic film, sealing, keeping the temperature at 30-35 ℃ (preferably, keeping the temperature at 30 ℃), fermenting for 10-20 days (preferably, fermenting for 10 days), performing solid-liquid separation after fermentation is finished, repeatedly fermenting the wormwood once under the same condition, combining two fermentation liquors, filtering and packaging to obtain the wormwood traditional Chinese medicine micro-ecological fermentation preparation, wherein the pH value is 3.0-3.5.
Preferably, the fermentation sterilization culture medium aqueous solution is prepared by the following method: adding 30g of corn steep liquor powder and 30g of molasses into each liter of liquid culture medium, and autoclaving at 121 deg.C for 15min to obtain the final product.
Preferably, the formulation of the wormwood traditional Chinese medicine micro-ecological fermentation preparation is water, powder or tablets.
The Chinese medicinal herb micro-ecological fermentation preparation of the invention is applied as a feed additive.
According to the wormwood traditional Chinese medicine micro-ecological fermentation preparation, the fermentation and conversion effects of the lactobacillus rhamnosus YL198 are mainly utilized, on one hand, the lactobacillus rhamnosus YL198 ferments wormwood, active ingredients in the wormwood are converted into medicinal ingredients beneficial to improving the growth performance of livestock and poultry and enhancing the immunity of the livestock and poultry, and the medicinal effect of the wormwood is improved; on the other hand, the lactobacillus rhamnosus YL198 is proliferated in a culture medium containing the wormwood, is metabolized to generate various functional substances and nutrient components such as bioactive enzymes, organic acids, amino acids, biological polysaccharides, vitamins and the like, and has the effects of promoting digestion of livestock and poultry and supplementing nutrition. Meanwhile, the magnetic field generated by the terahertz ore is used for assisting probiotic fermentation and conversion of the wormwood bioactive components during fermentation, so that the function of the wormwood lactobacillus microecological preparation product is further increased. Experiments prove that the Chinese medicinal herb microecological fermentation preparation has the functions of preventing and treating constipation and diarrhea, regulating intestinal flora, reducing the occurrence of diseases, improving the production performance and the like, and has the advantages of environmental protection, exact curative effect and capability of reducing or replacing the use of antibiotics
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The experimental procedures in the following examples are conventional unless otherwise specified.
The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified.
In the quantitative tests in the following examples, three replicates were set up and the results averaged.
Example 1 isolation, identification and preservation of strains
First, isolation of the Strain
Putting a naturally fermented wormwood sample into a transport culture medium, quickly putting the wormwood sample into an ice box, and conveying the wormwood sample to a laboratory for strain separation. Grinding and diluting the pickled Chinese cabbage sample in a gradient manner, separating lactobacillus in the sample by using a lactobacillus selective culture medium, culturing for 48h at 37 ℃, selecting typical bacterial colonies (milky white, round, neat edges, convex, opaque and moist and smooth surfaces) on a flat plate, carrying out streak culture on an MRS solid culture medium, and separating to obtain pure bacterial colonies. Inoculating the pure cultured strain to liquid MRS culture medium, culturing, adding 60% glycerol, and storing in refrigerator at-80 deg.C. Of these, 1 strain of Lactobacillus was designated YL 198.
II, identification of the strains
Physiological and biochemical identification results of YL 198: gram staining is positive, catalase test is negative, benzidine test is negative, indole test is negative, and acetyl methyl methanol test is positive; starch is not hydrolyzed, gelatin is not liquefied, hydrogen sulfide is not generated, and acid and gas are not generated by fermenting glucose; immotile bacilli; the growth can be carried out at 15 ℃ and 45 ℃, and the optimal growth temperature is 37-42 ℃; the pH is suitably 5.0 to 7.0; tolerates 6.5% NaCl; YL198 shows uniform turbid growth in liquid MRS medium, and the thallus is white precipitate after long-term storage.
The API 50CH (Merria, France) identification of YL198 is given in the following table:
in the above table, "+" indicates that the strain is a positive reaction; "-" indicates that the strain was negative.
16S rDNA sequence homology analysis: YL198 genome DNA is extracted, PCR amplification is carried out on the gene by utilizing lactobacillus primers (16 sF: 5 '-AGAGTTTGATCCTGGCTCAG-3'; 16 sR: 5 '-AGAAAGGAGGTGATCCAGC-3'), an amplification product is sent to the company Limited of biological engineering (Shanghai) to carry out sequencing, and the obtained 16s rDNA sequence is shown as a sequence 1 in a sequence table. Homology alignment analysis of the 16s rDNA sequence in GenBank database by BLAST program found that the homology of YL198 to Lactobacillus rhamnosus reaches 99%.
From the above results, the strain YL198 was identified as Lactobacillus rhamnosus (Lactobacillus rhamnosus).
Third, preservation of the Strain
The Lactobacillus rhamnosus (Lactobacillus rhamnosus) YL198 is preserved in China Center for Type Culture Collection (CCTCC) in 11 months and 27 days of 2020, and has the address as follows: eight-path Wuhan university school (Wuhan university collection center) 299 in Wuchang area, Wuhan city, Hubei province has the collection number: CCTCC NO: m2020781.
Example 2 preparation of a Chinese medicinal micro-ecological fermented preparation of Artemisia argyi of the present invention
Preparation of zymocyte suspension
Inoculating Lactobacillus rhamnosus YL198 (hereinafter referred to as YL 198) in liquid MRS culture medium, standing at 37 deg.C for 16 hr, centrifuging at 4 deg.C and 6000rpm for 8min, and collectingCollecting thallus precipitate, suspending with fermentation sterilization culture medium aqueous solution, adjusting viable count to 1.0 × 109CFU/ml to obtain YL198 bacterial suspension; wherein, the aqueous solution of the fermentation and sterilization culture medium is prepared by the following method: adding 30g of corn steep liquor powder and 30g of molasses into each liter of liquid culture medium, and autoclaving at 121 ℃ for 15 min.
Second, preparation of Chinese medicine micro-ecological fermented preparation of wormwood
Naturally drying the overground part of fresh wormwood, soaking for 2 times by using 1.75% of edible salt water, cleaning by using clear water, putting terahertz ore at the bottom of a fermentation tank, putting the wormwood, covering the wormwood with the terahertz ore, adding YL198 bacterial suspension, wherein the total weight of the terahertz ore is 1.5% of the total weight of the wormwood, adding the YL198 bacterial suspension and the wormwood according to the volume ratio of 1L to 300kg, compacting, covering a plastic film, sealing, fermenting at the constant temperature of 30 ℃ for 10 days, performing solid-liquid separation after the fermentation is finished, fermenting the wormwood again under the same condition, combining two fermentation liquids, filtering and packaging to obtain the wormwood traditional Chinese medicine micro-ecological fermentation preparation.
After fermentation is finished, the content of fermentation thalli in the obtained traditional Chinese medicine fermentation product is not less than 2.5 hundred million/ml, and the pH value is 3.0-3.5. The wormwood probiotic fermentation product, namely the wormwood traditional Chinese medicine micro-ecological fermentation preparation, obtained in the embodiment can be directly used as a feed additive.
Example 3 influence of the Chinese herbal medicine micro-ecological fermented preparation of wormwood on growth performance and serum biochemical indexes of weaned pigs
1. The experiment selects 40 weaned piglets with similar age in days and weight and good health condition, and randomly divides the weaned piglets into a control group and an experimental group, wherein each group has 2 repetitions, and each repetition has 10 piglets. After weaning at 21 days, in addition to normal feed feeding, the experimental group was fed with mugwort Chinese medicinal micro-ecological fermentation preparation (content: 3%), and the control group was fed with normal feed and free drinking water only, without mugwort Chinese medicinal micro-ecological fermentation preparation, and fed continuously for 30 days.
2. Index of growth performance
The daily feed intake of piglets was recorded and the weight of each duplicate piglet was weighed at 21 and 51 days of age. Average Daily Feed Intake (ADFI), Average Daily Gain (ADG) and feed to weight ratio (F/G) were calculated. The results are shown in Table 1.
TABLE 1
Indicates significant difference (P <0.05) compared to the control group, and indicates very significant difference (P <0.01) compared to the control group.
As can be seen from Table 1, the average daily food intake was slightly lower in the experimental group than in the control group, but the difference between the two groups was not significant (P > 0.05); the average daily gain of the experimental group is obviously higher than that of the control group; the material weight ratio of the experimental group is obviously lower than that of the control group. Therefore, the Chinese medicinal herb microecological fermentation preparation of wormwood can obviously improve the growth performance of piglets.
3. Determination of diarrhea Rate
The number of diarrhea heads per repeat per day was recorded and the diarrhea rate was calculated. The diarrhea rate (%) × 100 [ total diarrhea per replicate piglet head/total head per replicate × number of trial days) ]. As a result, the diarrhea rate of the control group was 3.34%, while the diarrhea rate of the experimental group was significantly lower than that of the control group, only 1.46%. Therefore, the wormwood traditional Chinese medicine micro-ecological fermentation preparation can obviously reduce the diarrhea rate of piglets.
4. Determination of serum biochemical indicators
When piglets are 33 days old, 1 head of normal pigs are randomly taken from each repetition, 10mL of blood is collected from the anterior vena cava, serum is centrifugally separated, and the piglets are frozen and preserved at the temperature of 20 ℃ below zero, and the total serum protein (TP), Albumin (ALB), Globulin (GLB), glucose, Urea Nitrogen (UN), immunoglobulin G (IgG), immunoglobulin M (IgM) and immunoglobulin A (IgA) contents, glutamic-pyruvic transaminase (GTP) and glutamic-oxalacetic transaminase (GOT) activities are measured. Serum indexes are measured by a full-automatic biochemical analyzer. The results are shown in Table 2.
TABLE 2
Indicates significant difference (P <0.05) compared to the control group, and indicates very significant difference (P <0.01) compared to the control group.
As can be seen from Table 2, the serum total protein and albumin contents of the experimental group are significantly higher than those of the control group, and the difference between the serum globulin contents of the experimental group and those of the control group is very significant, so that the strong synthesis capacity of tissue protein can be demonstrated, and the growth of tissue organs can be promoted, therefore, the serum protein content is an important index for measuring the animal nutrition status. The content can accurately reflect the balance condition between protein metabolism and amino acid in the animal body, the serum urea nitrogen content of an experimental group is lower than that of a control group, and the lower serum urea nitrogen content shows that the amino acid balance is good and the protein synthesis rate of the organism is higher. The IgG content of the experimental group is obviously higher than that of the control group, which shows that the wormwood Chinese medicinal micro-ecological fermentation preparation has a promotion effect on the development of the immune system of piglets and is beneficial to activating the humoral immune response of organisms.
Example 4 Effect of Chinese mugwort herb micro-ecological fermented preparation of the present invention on the immune function of chickens
1. Selecting 60 feathers of 14-day-old Specific Pathogen Free (SPF) chicks, randomly dividing the feathers into 2 groups, each group having 5 repetitions and each repetition having 6 feathers. The control group is fed with basic feed and free drinking water without adding Chinese medicinal microecological fermentation preparation of folium Artemisiae Argyi, the experimental group is fed with basic feed and free drinking water with Chinese medicinal microecological fermentation preparation of folium Artemisiae Argyi (content of 1%) added into drinking water, and fed for 21d continuously.
2. Measurement of growth Performance
The chickens were weighed on test days 1 and 21, weighed as feed, and the Average Daily Feed Intake (ADFI), Average Daily Gain (ADG) and feed-to-weight ratio (F/G) during the test period were calculated in units of replicates. The results are shown in Table 3.
TABLE 3
Indicates significant difference (P <0.05) compared to the control group, and indicates very significant difference (P <0.01) compared to the control group.
As can be seen from Table 3, the average daily food intake of the experimental group is slightly lower than that of the control group, but the difference between the two groups is not significant (P is more than 0.05), the average daily gain of the experimental group is significantly higher than that of the control group, and the material weight ratio of the experimental group is significantly lower than that of the control group. Therefore, the Chinese medicinal herb microecological fermentation preparation of wormwood can obviously improve the growth performance of the chicks.
3. Antibodies and cytokines
Randomly taking 1 feather chicken in each repetition of each group on the 21 st day of the experiment, aseptically collecting blood from heart, standing at 4 deg.C for 1h, centrifuging at 3000r/min for 15min, separating serum, and storing at-20 deg.C. Measuring IgA, IgM, IgG, IL-1, IL-2, IL-4 and IL-6 contents in chick serum by enzyme-linked immunosorbent assay. The results are shown in tables 4 and 5.
TABLE 4
Indicates significant difference (P <0.05) compared to the control group, and indicates very significant difference (P <0.01) compared to the control group.
As can be seen from Table 4, the levels of serum cytokines IL-2, IL-6 and IL-1 in the chicks in the experimental group are significantly lower than those in the control group, so that the wormwood Chinese medicinal micro-ecological fermentation preparation has the effect of reducing the serum inflammation of the chicks.
TABLE 5
| Group of | IgA(ng/ml) | IgM(ng/ml) | IgG(ng/ml) |
| Control group | 121.36±6.33 | 432.01±10.23 | 633.98±14.01 |
| Experimental group | 134.29±7.25* | 496.32±9.88* | 788.21±13.02** |
Indicates significant difference (P <0.05) compared to the control group, and indicates very significant difference (P <0.01) compared to the control group.
As can be seen from Table 5, the contents of IgA and IgM in the serum of the experimental group chicks are both significantly higher than those of the control group, the content of IgG in the serum of the experimental group chicks is significantly higher than that of the control group, and the difference is very significant (P <0.01), so that the wormwood traditional Chinese medicine micro-ecological fermentation preparation disclosed by the invention can activate the immune system of the chicks.
Example 5 influence of Chinese medicinal herb micro-ecological fermented preparation of Artemisia argyi on production performance and meat quality of fattening sheep
1. 20 Duhan hybrid F1 mutton sheep weighing about 20kg were selected and randomly divided into control and experimental groups, each group consisting of 10 sheep (half male and half female), and the two groups were separately bred. In addition to normal compound feed feeding, the experimental group was continuously fed with 1.5% Chinese medicinal herb microecological fermentation preparation, and the control group was fed with free drinking water and no Chinese medicinal herb microecological fermentation preparation for 100 days.
2. Daily gain and feed conversion ratio
The daily gain adopts the method of respectively measuring the initial weight and the final weight of a test sheep in an empty stomach in the morning, calculating the average daily gain, recording the average daily consumption of the feed of each group during the test period, and calculating the feed reward. The results are shown in Table 6.
TABLE 6
Indicates significant difference (P <0.05) compared to the control group, and indicates very significant difference (P <0.01) compared to the control group.
As can be seen from table 6, the average daily gain of the experimental group was significantly higher than that of the control group. Under the condition that the average daily feed intake difference is not obvious, the feed-meat ratio of the experimental group is obviously lower than that of the control group, so that the wormwood Chinese medicinal micro-ecological fermentation preparation obviously improves the production performance of fattening sheep.
3. Slaughter and abdominal fat rates
The dressing percentage is the percentage of the total weight of ketone bodies to the total weight of the living body before slaughtering, and the abdominal fat percentage is the percentage of the total weight of abdominal fat to the total weight of the living body. The results of the dressing percentage and abdominal fat percentage calculations are shown in table 7.
TABLE 7
| Group of | Live weight of slaughter/kg | Hot carcass weight/kg | Slaughter Rate/% | Weight of hot abdominal fat/kg | Abdominal fat ratio-％ |
| Control group | 38.65±1.54 | 25.03±0.99 | 64.76±1.35 | 1.10±0.32 | 2.84±0.12 |
| Experimental group | 47.96±1.36** | 28.87±0.97* | 60.19±1.52 | 0.94±0.41* | 1.96±0.22* |
Indicates significant difference (P <0.05) compared to the control group, and indicates very significant difference (P <0.01) compared to the control group.
As can be seen from table 7, in each index of slaughter performance, the final slaughter live weight test group is higher than the control group under the condition that the initial weight is basically the same, and the difference is very significant (P < 0.01); the hot carcass weight test group is improved compared with the control group, and the difference is obvious (P <0.05), but the difference of the dressing percentage is not obvious (P > 0.05). The decrease of the abdominal fat rate of the experimental group compared with the control group effectively shows that the feeding of the wormwood Chinese medicinal micro-ecological fermentation preparation can obviously reduce the deposition of the abdominal fat.
4. Influence of wormwood traditional Chinese medicine micro-ecological fermentation preparation on quality of fattening mutton
The marbling grade at the cross section of the eye muscle is determined by comparing the marbling grade pictures, the marbling is divided into 1 grade, 1.5 grade, 2 grade, 2.5 grade, 3 grade, 3.5 grade and 4 grade, the marbling is extremely rich in 1 grade, the marbling is rich in 2 grade, the marbling is little in 3 grade, the marbling is almost not in 4 grade, and the marbling is 0.5 grade between the two grades. The meat color grade is divided into 1-8 grades according to the color depth, and the best grade is 3-5 grades. The pH was measured using a direct measurement carcass pH meter, at 45min and 24h after slaughter. The water loss rate is an evaluation index of intramuscular protein change before and after mutton slaughter, and directly influences freshness, flavor and color of mutton. The cooked meat rate is a measure of the weight loss of mutton after heating at a certain temperature for a certain period of time. The influence of the Chinese medicinal herb micro-ecological fermentation preparation on the quality of fattening mutton is determined according to the prior art. The results are shown in Table 8.
TABLE 8
Indicates significant difference (P <0.05) compared to the control group, and indicates very significant difference (P <0.01) compared to the control group.
As can be seen from Table 8, the marbling of the experimental group was superior to that of the control group, and there was no significant difference between the flesh-colored groups. The results of measuring the pH value between the longissimus dorsi of fattening sheep slaughtered for 45min and the pH value between the longissimus dorsi of fattening sheep stored for 24h at 4 ℃ respectively show that the pH values of the two groups are reduced to different degrees along with the time extension, and the phenomenon can be caused by that after slaughtering, the blood circulation stops, muscle cells are in an anaerobic state, and the in vivo glycogen is used to generate lactic acid, so that the pH value of meat is reduced. For the cooked meat rate, the difference between the two groups was not significant (P > 0.05). The water loss rate reflects the degree of intramuscular protein change and intramuscular water loss after slaughter, and the water loss rate directly influences the tenderness, flavor, color and the like of mutton. The water loss rate of the experimental group is obviously lower than that of the control group, the lower the water loss rate is, the stronger the water retention performance is, and the tender the meat quality is, so that the wormwood Chinese medicinal micro-ecological fermentation preparation can improve the water retention performance of the fattening mutton.
The invention discloses a wormwood traditional Chinese medicine micro-ecological fermentation preparation and a preparation method and application thereof, and a person skilled in the art can realize the preparation by properly improving process parameters by referring to the content. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the invention has been described in terms of preferred embodiments, it will be apparent to those skilled in the art that the technology can be practiced and applied by modifying or appropriately combining the products described herein without departing from the spirit and scope of the invention.
Sequence listing
<110> Shanghai plant fermentation Living Biotech Co., Ltd
<120> Chinese mugwort herb micro-ecological fermentation preparation and preparation method and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1310
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
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ctcaccaagg caatgatacg tagccgaact gagaggttga tcggccacat tgggactgag 180
acacggccca aactcctacg ggaggcagca gtagggaatc ttccacaatg gacgcaagtc 240
tgatggagca acgccgcgtg agtgaagaag gctttcgggt cgtaaaactc tgttgttgga 300
gaagaatggt cggcagagta actgttgtcg gcgtgacggt atccaaccag aaagccacgg 360
ctaactacgt gccagcagcc gcggtaatac gtaggtggca agcgttatcc ggatttattg 420
ggcgtaaagc gagcgcaggc ggttttttaa gtctgatgtg aaagccctcg gcttaaccga 480
ggaagtgcat cggaaactgg gaaacttgag tgcagaagag gacagtggaa ctccatgtgt 540
agcggtgaaa tgcgtagata tatggaagaa caccagtggc gaaggcggct gtctggtctg 600
taactgacgc tgaggctcga aagcatgggt agcgaacagg attagatacc ctggtagtcc 660
atgccgtaaa cgatgaatgc taggtgttgg agggtttccg cccttcagtg ccgcagctaa 720
cgcattaagc attccgcctg gggagtacga ccgcaaggtt gaaactcaaa ggaattgacg 780
ggggcccgca caagcggtgg agcatgtggt ttaattcgaa gcaacgcgaa gaaccttacc 840
aggtcttgac atcttttgat cacctgagag atcaggtttc cccttcgggg gcaaaatgac 900
aggtggtgca tggttgtcgt cagctcgtgt cgtgagatgt tgggttaagt cccgcaacga 960
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Claims (7)
1. The microecological fermentation preparation of the Chinese mugwort herb is characterized by comprising a Lactobacillus rhamnosus YL198 bacterial suspension and the mugwort herb, wherein the dosage ratio of the Lactobacillus rhamnosus YL198 bacterial suspension to the mugwort herb is as follows: the weight of the powder is 1L:100 kg-1L: 500 kg;
the Lactobacillus rhamnosus (Lactobacillus rhamnosus) YL198 is preserved in China center for type culture Collection at 11 month and 27 month in 2020, and the preservation number is as follows: CCTCC NO: m2020781.
2. The microecological fermented preparation of Artemisia argyi traditional Chinese medicine of claim 1, wherein the dosage ratio of Lactobacillus rhamnosus YL198 bacterial suspension to Artemisia argyi is: the volume and weight are 1L to 300 kg.
3. The preparation method of the wormwood traditional Chinese medicine micro-ecological fermentation preparation according to claim 1, comprising the following steps:
inoculating lactobacillus rhamnosus YL198 to a liquid MRS culture medium, performing static culture at 37-45 ℃ for 16-25 h, centrifuging at 4-10 ℃ and 4000-8000 rpm for 5-10 min, and collecting thallus precipitates; suspending with aqueous solution of fermentation and sterilization culture medium, adjusting viable count to 1.0 × 109～1.0×1010CFU/ml to obtain YL198 bacterial suspension;
naturally drying the overground part of fresh wormwood, soaking for 2 times by using 0.5-1% of saline solution, cleaning with clear water, placing terahertz ore at the bottom of a fermentation tank, placing the wormwood, covering the wormwood with the terahertz ore, wherein the total weight of the terahertz ore is 1-2% of the total weight of the wormwood, adding YL198 bacterial suspension, covering a plastic film, sealing, fermenting for 10-20 days at constant temperature of 30-35 ℃, performing solid-liquid separation after fermentation is finished, fermenting the wormwood again under the same condition, combining fermentation liquor obtained in two times, filtering, and packaging to obtain the wormwood traditional Chinese medicine micro-ecological fermentation preparation, wherein the pH value is 3.0-3.5.
4. The method of claim 3, comprising the steps of:
inoculating Lactobacillus rhamnosus YL198 to liquid MRS culture medium, standing at 37 deg.C for 16 hr, centrifuging at 4 deg.C and 6000rpm for 8min, and collecting thallus precipitate; suspending with aqueous solution of fermentation and sterilization culture medium, adjusting viable count to 1.0 × 109CFU/ml to obtain YL198 bacterial suspension;
naturally drying the overground part of fresh wormwood, soaking for 2 times by using 0.75% of edible salt water, cleaning by using clear water, putting terahertz ore at the bottom of a fermentation tank, putting the wormwood, covering the wormwood with the terahertz ore, adding YL198 bacterial suspension, wherein the total weight of the terahertz ore is 1.5% of the total weight of the wormwood, adding the YL198 bacterial suspension and the wormwood according to the volume ratio of 1L to 300kg, covering a plastic film, sealing, carrying out constant temperature of 30 ℃, fermenting for 10 days, carrying out solid-liquid separation after the fermentation is finished, carrying out re-fermentation on the wormwood once under the same condition, combining two fermentation liquids, filtering and packaging to obtain the wormwood traditional Chinese medicine micro-ecological fermentation preparation, wherein the pH value is 3.0-3.5.
5. The method according to claim 3, wherein the aqueous solution of the fermentative sterilization medium is prepared by the following method: adding 30g of corn steep liquor powder and 30g of molasses into each liter of liquid culture medium, and autoclaving at 121 deg.C for 15min to obtain the final product.
6. The preparation method according to claim 3, wherein the mugwort Chinese herbal micro-ecological fermentation preparation is in the form of aqua, powder or tablet.
7. The use of the wormwood Chinese medicinal micro-ecological fermented preparation according to claim 1 or 2 as a feed additive.
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