CN113481110B - Preparation method and application of Auricularia polytricha selenium-rich culture - Google Patents
Preparation method and application of Auricularia polytricha selenium-rich culture Download PDFInfo
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- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 title claims abstract description 82
- 229910052711 selenium Inorganic materials 0.000 title claims abstract description 82
- 239000011669 selenium Substances 0.000 title claims abstract description 82
- 240000005710 Auricularia polytricha Species 0.000 title claims abstract description 37
- 235000000024 Auricularia polytricha Nutrition 0.000 title claims abstract description 37
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 241000321977 Auricularia fuscosuccinea Species 0.000 claims abstract description 36
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- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims 2
- 239000002609 medium Substances 0.000 claims 2
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- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims 1
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- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims 1
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Abstract
The invention discloses a preparation method and application of a Auricularia polytricha selenium-rich culture, which comprises the following steps: step one, preparing a liquid culture medium of the Auricularia polytricha selenium-rich culture; step two, preparing a Auricularia polytricha selenium-rich culture solid culture medium; inoculating a Auricularia fuscospora (Auricularia cornea) in an Auricularia fuscospora liquid culture medium, and fermenting for a preset time to obtain a cultured Auricularia fuscospora liquid strain; and step four, inoculating the cultured liquid strain of the Auricularia fuscosuccinea, mixing the inoculated liquid strain with a solid culture medium, fermenting the mixture for a preset time, and drying the mixture at a low temperature to obtain the selenium-rich Auricularia fuscosuccinea culture. The prepared Auricularia polytricha selenium-rich culture can increase the feed-weight ratio of pigs, improve the oxidation resistance of pork, improve the selenium content of pork muscles and improve the oxidation stability of the pork.
Description
Technical Field
The invention belongs to the field of feeds, and particularly relates to a preparation method and application of a Auricularia polytricha selenium-rich culture (feed).
Background
Selenium-enriched feed is usually added in the animal breeding process. So that the meat of animals, such as pork, is rich in selenium element to supplement selenium in human body. For example, patent 201810432340.X discloses a selenium-rich pork and a method for feeding selenium-rich pigs. However, feeding pigs with the selenium-rich feed may cause negative effects on pig growth, such as slow pig growth, pork quality and the like, so that the development of the selenium-rich feed, determination of the effect on pig growth, namely quality, and determination of the dosage are needed.
Disclosure of Invention
In order to solve the problems, the invention discloses a preparation method and application of a Auricularia fuscosuccinea selenium-rich culture. The prepared Auricularia polytricha selenium-rich culture can increase the feed-weight ratio of pigs, improve the oxidation resistance of pork, improve the selenium content of pork muscles and improve the oxidation stability of pork.
In order to realize the purpose, the technical scheme of the invention is as follows:
a preparation method of a Auricularia polytricha selenium-rich culture comprises the following steps:
step one, preparing a selenium-rich culture liquid culture medium of Auricularia fuscosuccinea;
step two, preparing a Auricularia polytricha selenium-rich culture solid culture medium;
inoculating a Auricularia aurea mycelium (Auricularia cornea) into the Auricularia aurea selenium-rich culture liquid culture medium, and fermenting for a preset time to obtain a cultured Auricularia aurea liquid strain;
and step four, inoculating the cultured Auricularia fuscosuccinea liquid strain, mixing the inoculated Auricularia fuscosuccinea liquid strain with a solid culture medium of Auricularia fuscosuccinea selenium-rich culture, fermenting for a preset time, and drying at a low temperature to obtain the Auricularia fuscosuccinea selenium-rich culture.
In a further improvement, the liquid culture medium of the selenium-rich culture of the Auricularia polytricha comprises, by mass, 0.003% of sodium selenite, 3% of glucose, 2% of corn flour, 40.25% of KH2PO40, 40.15% of MgSO15 and the balance water.
Further improvement, the preparation method of the Auricularia polytricha selenium-rich culture solid culture medium comprises the following steps: mixing a main material and an auxiliary material to obtain a mixture, wherein the main material comprises 85 mass percent of wheat bran and 15 mass percent of corn flour; the auxiliary material is sodium selenite, and the weight of the auxiliary material is 0.003 percent of that of the main material; sterilizing the mixture at 120 ℃ for 1 hour, introducing sterile water, and cooling to obtain the Auricularia fuscosuccinea selenium-enriched culture solid culture medium, wherein the fermentation water content in the Auricularia fuscosuccinea selenium-enriched culture solid culture medium is controlled at 60%.
In the third step, the inoculation amount of the fungus mycelium (Auricularia cornea) is 10g/L, and the fermentation conditions are as follows: fermenting at 25 deg.C and 180RPM for 7 days.
In the fourth step, the Auricularia fuscosuccinea liquid strain is mixed with the Auricularia fuscosuccinea selenium-rich culture solid culture medium in an inoculation amount of 10%, and then fermented for 15 days at the temperature of 27 ℃ and the relative humidity of 60%.
The application of the Auricularia fuscosuccinea selenium-rich culture is characterized in that the Auricularia fuscosuccinea selenium-rich culture is used as a feed additive for increasing the growth performance of pigs and the quality of pork.
In a further improvement, the Auricularia polytricha selenium-rich culture is used as a feed additive for increasing the weight ratio of pig feed, improving the oxidation resistance of pork, improving the selenium content of pork muscle and improving the oxidation stability of pork.
In a further improvement, the Auricularia fuscosuccinea selenium-rich feed is also used as a feed additive for improving the content of butyric acid in pork, improving the expression level of MCT1mRNA in the colon of a pig, improving the content of GSH-Px in the serum and the liver of the pig and reducing the content of MDA in the liver.
In a further improvement, the addition amount of the Auricularia polytricha selenium-rich culture is 1.2-2.4% of the total feed weight.
The invention has the advantages that:
the prepared Auricularia polytricha selenium-rich culture can increase the feed-weight ratio of pigs, improve the oxidation resistance of pork, improve the selenium content of pork muscles and improve the oxidation stability of pork.
Drawings
Figure 1 is a graph of the relative expression of MCT1mRNA in colon in pigs fed a control diet and supplemented with 0.6%, 1.2% and 2.4% (SAC) diets, respectively.
Detailed Description
The invention is further explained by the following embodiments in conjunction with the drawings.
Example 1
The preparation method of the Auricularia polytricha selenium-rich culture comprises the following steps:
preparing a liquid culture medium of the Auricularia polytricha selenium-rich culture: 0.003 percent of sodium selenite, 3 percent of glucose, 2 percent of corn flour and KH 2 PO 4 0.25％，MgSO 4 0.15 percent. Then inoculating 10g/L Auricularia (Auricularia cornea) mycelium, and fermenting at 25 deg.C and 180RPM for 7 days.
Preparing a solid culture medium: wheat bran 85%, corn flour 15%, and sodium selenite 0.003%. Sterilizing at 120 deg.C for 1 hr, introducing sterile water, and cooling. The fermentation moisture is controlled at 60%.
The cultured Auricularia fuscosuccinea liquid strain is mixed with solid culture medium at an inoculum size of 10%, and fermented at 27 deg.C and 60% relative humidity for 15 days.
Then drying the culture by low-temperature drying equipment (the temperature is controlled to be less than or equal to 55 ℃) to obtain the finished product of the Auricularia polytricha selenium-rich culture. Detecting and analyzing nutrient substances of the Auricularia polytricha selenium-rich culture (SAC).
TABLE 1 nutrient analysis of Auricularia polytricha selenium-enriched culture
Auricularia fuscosuccinea selenium-rich culture
1. The selenium-rich culture of the Auricularia fuscosuccinea is used for the growth fattening feeding test:
the 96-head ternary hybridization growth fattening pig is randomly divided into 4 treatments, and each treatment is 6 times repeated. The 4 treatments were fed a control diet and a test diet supplemented with 0.6%, 1.2% and 2.4% selenium-enriched culture of Auricularia fuscosuccinea, respectively. All diets met the nutritional requirements of NRC (2012). The test period was 45 days. The ambient temperature is controlled at 25-28 ℃. The test is divided into two stages, a growth stage (stage 1: 0-23 days) and a fattening stage (stage 2: 24-45 days). Daily feed intake and body weight were weighed and recorded on days 0, 23 and 45 of the trial, respectively, and the Average Daily Gain (ADG), average Daily Feed Intake (ADFI) and feed-meat ratio (F/G) were determined. And on 45 days, taking a serum sample and a fresh excrement sample for detecting serum biochemical indexes and short-chain fatty acids.
At the end of the test, 24 pigs (1 per pen, 6 per treatment) were selected for slaughter testing. After fasting (12 h), slaughter was performed. Colonic tissue specimens were taken and stored at-80 ℃ for mRNA expression analysis. The tissues of the 10 th to 12 th intercostal longissimus muscle (LDM) (about 200 g), liver and kidney of the right half of carcass were collected and immediately stored at-20 ℃ for meat quality evaluation.
TABLE 2 test diet composition
Chemical analysis and calculation:
samples were tested for crude fat (EE), dry Matter (DM), crude Protein (CP), ash, total phosphorus and calcium. Neutral Detergent Fiber (NDF) and Acid Detergent Fiber (ADF) contents were determined using a fiber analyzer (2010, foss, denmark). Total energy (GE) in diet, faeces and urine was measured using an oxygen bomb calorimeter (C2000, IKA, germany). The selenium content was quantified using inductively coupled plasma spectroscopy (ICP-MS, thermo, USA). The concentration of Short Chain Fatty Acids (SCFA) was analyzed by ion chromatography (883, metrohm, switzerland) according to Wu et al (2017). A commercial detection kit is adopted to detect the oxidation resistance indexes of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), malondialdehyde (MDA) and total oxidation resistance (T-AOC). Serum High Density Lipoprotein (HDL), low Density Lipoprotein (LDL), glucose (GLU), total Cholesterol (TC), and Triglyceride (TG) levels were determined using a Hitachi 3100 full-automatic biochemical analyzer, and a related kit.
Carcass traits and meat quality determination:
the method for measuring the length of the carcass, the weight of the carcass, marbling, eye muscle area, backfat thickness, hot carcass weight and the like refers to the technical specification NY/T825-2004 for measuring the carcass traits of lean-type pigs
Meat quality indicators, including meat color, pH, shear, cooking loss and drip loss, were then examined.
Relative expression amount of MCT1mRNA expression: the expression level of mRNA of the monocarboxylate transporter 1 (MCT 1) was analyzed by RT-PCR. MCT1 and beta-actin as shown in Table 3.
TABLE 3 primer information
Fresh meat oxidation stability at simulated retail display:
24 fresh meat samples were selected. Then, the fresh meat with thickness of 2.5 cm, no bone and neatly cut is respectively put on a foamed plastic tray and then wrapped by a breathable polyvinyl chloride film. Subsequently, all packaged samples were stored in an open top display box, temperature maintained at 4 ℃ and continuous lamp illumination conditions (TLD-T836W, 6200K). On days 0, 3 and 6, samples of about 10g were taken for MDA content analysis.
And (3) data analysis:
the growth performance of the raw meat, the biochemical indexes of serum, short chain fatty acids, the quality of the meat, the selenium concentration of the tissue and the content of MDA are analyzed by adopting SAS (SAS Inst.Inc., USA) software version 9.2. P <0.05 was considered statistically significant.
And (3) test results:
TABLE 4 influence of Auricularia polytricha selenium culture on growth performance of growing-finishing pigs
As shown in table 4, the feed weight ratio (p < 0.05) for pigs with SAC addition was significantly lower than the control on days 1-23 of the experiment. On days 24-45 of the test, the weight ratio of the swine feed with 1.2% and 2.4% SAC added was significantly higher than that of the control and with 0.6% SAC added (p < 0.01). The average daily gain was higher for 1.2% and 2.4% SAC treatment than the control (p < 0.05), 0.6%, 1.2% and 2.4% SAC treated less than the control (p < 0.05) throughout the test period.
As can be seen from table 5, addition of SAC to the ration significantly increased the butyric acid concentration for 45 days (p < 0.05) compared to the control group.
TABLE 5 Effect of Auricularia fuscosuccinea selenium-rich culture on short-chain fatty acids in fresh feces
FIG. 1 Effect of selenium-enriched culture of Auricularia fuscosuccinea on expression level of MCT1mRNA in colon
As shown in figure 1, the porcine colon MCT1mRNA expression (P = 0.04) was higher for 1.2% and 2.4% sac diets than for the control group.
TABLE 6 influence of selenium-enriched Auricularia polytricha culture on serum biochemical index
As can be seen from Table 6, the GSH-Px content was significantly higher on day 45 in the 1.2% and 2.4% SAC groups than in the control group (p < 0.05).
TABLE 7 Effect of selenium-enriched Auricularia polytricha culture on antioxidant index of liver and longissimus dorsi
As can be seen from table 7, the addition of 2.4% sac to the diet significantly increased the hepatic GSH-Px content (p < 0.05), and the addition of 1.2% and 2.4% sac significantly decreased the hepatic MDA content (p < 0.05). GSH-Px activity was significantly higher in the 2.4% treated group than in the control group (p < 0.05).
TABLE 8 Effect of Auricularia fuscosuccinea selenium-enriched cultures on tissue selenium content
As can be seen from Table 8, the selenium content in the LDM for the fattening pig added with 2.4% SAC was higher (p < 0.05) than that in the other 3 treatments. The selenium content of the liver of the pigs fed with SAC daily ration tends to increase (p = 0.09).
TABLE 9 Effect of selenium-enriched Auricularia polytricha culture on carcass traits and meat quality
As can be seen from Table 9, the feeding of SAC at different levels had no effect on the carcass traits and meat quality of the swine.
TABLE 10 influence of selenium-enriched Auricularia fuscosuccinea culture on antioxidant status of fresh meat in simulated shelf life test
As can be seen from table 10, the simulated retail display had a lower fresh meat MDA content (p < 0.05) in the 2.4% sac group compared to the control group at day 6.
In conclusion, the Auricularia fuscosuccinea selenium-rich culture has good effects on promoting the growth and development of pigs, improving the oxidation resistance and the content of selenium in muscles and improving the oxidation stability of meat.
While embodiments of the invention have been described above, and not limited to the applications set forth in this specification and the embodiments, it is to be understood that the invention is capable of use in various other respects, all without departing from the general concept as defined in the appended claims and their equivalents, and further modifications may be readily made by those skilled in the art, and it is therefore not to be limited to the specific details and illustrations contained herein.
Claims (6)
1. The application of the Auricularia polytricha selenium-rich culture is characterized in that the Auricularia polytricha selenium-rich culture is used as a feed additive for increasing the growth performance of pigs and the quality of pork; the preparation method of the Auricularia polytricha selenium-rich culture comprises the following steps: step one, preparing a selenium-rich culture liquid culture medium of Auricularia fuscosuccinea;
step two, preparing a Auricularia polytricha selenium-rich culture solid culture medium;
step three, inoculating the Auricularia fuscosuccinea mycelium in the Auricularia fuscosuccinea selenium-rich culture liquid culture mediumAuricularia cornea) Fermenting for a preset time to obtain a cultured Auricularia fuscosuccinea liquid strain;
inoculating the cultured liquid strain of the Auricularia fuscosuccinea, mixing the inoculated liquid strain with a solid culture medium, fermenting for a preset time, and drying at a low temperature to obtain an Auricularia fuscosuccinea selenium-rich culture; the liquid culture medium of the Auricularia polytricha selenium-rich culture comprises, by mass, 0.003% of sodium selenite, 3% of glucose, 2% of corn flour, 0.25% of KH2PO4, 0.15% of MgSO4 and the balance of water;
the preparation method of the Auricularia polytricha selenium-rich culture solid culture medium comprises the following steps: mixing a main material and an auxiliary material to obtain a mixture, wherein the main material comprises 85 mass percent of wheat bran and 15 mass percent of corn flour; the auxiliary material is sodium selenite, and the weight of the auxiliary material is 0.003 percent of that of the main material; sterilizing the mixture at 120 ℃ for 1 hour, introducing sterile water, and cooling to obtain the Auricularia fuscosuccinea selenium-rich culture solid medium, wherein the fermentation water content in the Auricularia fuscosuccinea selenium-rich culture solid medium is controlled at 60%.
2. The use of the Auricularia fuscosuccinea selenium-enriched culture of claim 1, wherein the Auricularia fuscosuccinea selenium-enriched culture is used as a feed additive for reducing the weight ratio of pig feed, improving the oxidation resistance of pork, improving the selenium content of pork muscle and improving the oxidation stability of pork.
3. The use of the Auricularia polytricha selenium-enriched culture of claim 1, wherein the Auricularia polytricha selenium-enriched feed is further used as a feed additive for increasing the content of butyric acid in pork, increasing the expression level of MCT1mRNA in the colon of a pig, increasing the content of GSH-Px in the serum and liver of a pig and reducing the content of MDA in the liver.
4. The use of the Auricularia polytricha selenium-enriched culture of claim 1, wherein the Auricularia polytricha selenium-enriched culture is added in an amount of 1.2-2.4% of the total feed weight.
5. The use of the selenium-enriched culture of Auricularia fuscosuccinea as claimed in claim 1, wherein in step three, the mycelium of Auricularia fuscosuccinea (L.), (Auricularia cornea) The inoculation amount of (2) is 10g/L, and the fermentation conditions are as follows: fermenting at 25 deg.C and 180RPM for 7 days.
6. The use of the Auricularia polytricha selenium-enriched culture of claim 1, wherein in the fourth step, the Auricularia polytricha liquid strain is mixed with the Auricularia polytricha selenium-enriched culture solid culture medium at an inoculation amount of 10%, and then fermented for 15 days at 27 ℃ and 60% relative humidity.
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