CN114681496B - Application of Acremonium terricola culture - Google Patents
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Abstract
The present disclosure belongs to the field of microorganisms, disclosing the use of a culture of Acremonium terrestris for non-therapeutic purposes, in particular to the use of a culture of Acremonium terrestris for increasing the number of Clostridium praecox in the gastrointestinal tract of an animal; use of a culture of acremonium for ameliorating inflammation in the gastrointestinal tract of an animal.
Description
Technical Field
The present disclosure is in the field of microorganisms, in particular to the use of Acremonium acutum cultures for increasing the number of Clostridium praecox in the gastrointestinal tract of animals; use of a culture of acremonium for ameliorating inflammation in the gastrointestinal tract of an animal.
Background
The clostridium praecox has a related relation with pathological changes of inflammatory bowel patients, the reduction of the clostridium praecox has a positive relation with the pathological changes of inflammatory bowel patients, the clostridium praecox can up-regulate TJ protein expression to reduce permeability of intestinal mucosa, an epithelial anaerobic barrier can be constructed to protect the intestinal mucosa, and butyric acid can be produced to reduce intestinal inflammation.
Disclosure of Invention
In view of this, the present application provides in a first aspect the use of a culture of Acremonium to enhance Clostridium praecox in the gastrointestinal tract of an animal, said use being for non-therapeutic purposes, to ameliorate inflammation in the gastrointestinal tract of an animal.
In view of this, the present application provides in a second aspect the use of a culture of Acremonium in a feed additive or medicament for non-therapeutic purposes, the feed additive being for increasing the elevation of Clostridium praecox in the gastrointestinal tract of an animal.
In view of this, the present application provides in a third aspect the use of a culture of Acremonium for the amelioration of inflammation in the gastrointestinal tract of an animal, said use for non-therapeutic purposes, said culture of Acremonium for use in the improvement of Clostridium praecox in the gastrointestinal tract of an animal, for the amelioration of inflammation in the gastrointestinal tract of an animal.
Optionally, according to the use of the first, second and third aspects, the animal is a chicken or pig.
Optionally, according to the application of the first aspect, the second aspect and the third aspect, the acremonium terrestris culture contains the following active substances, wherein the content of the active substances is: 0.001-5% of adenosine, 0.002-4% of ergosterol, 0.002-4% of guanosine, 0.002-4% of uridine and 0.002-10% of cordyceps polysaccharide.
Optionally, according to the use of the first, second and third aspects, the Acremonium culture is obtained by liquid fermentation or solid fermentation.
Optionally, according to the use of the first, second and third aspects, the acremonium terrestris culture is fermented by a liquid comprising the steps of:
the method comprises the steps of inoculating Acremonium to a test tube culture medium for culture, inoculating Acremonium after culture of the test tube strain culture medium to a shake flask culture medium for culture, inoculating Acremonium after culture of the shake flask culture medium to a fermentation tank culture medium for culture, pumping out fermentation liquor after culture of the fermentation tank strain, centrifuging, drying and crushing to form Acremonium culture.
Optionally, according to the use of the first, second and third aspects, the acremonium terrestris culture comprises the following steps by solid fermentation:
the method comprises the steps of inoculating Acremonium to a test tube strain culture medium for culture, inoculating Acremonium after culture in the test tube strain culture medium to a shake flask strain culture medium for culture, inoculating Acremonium after culture in the shake flask strain culture medium to a fermentation tank strain culture medium for culture, inoculating the fermentation tank strain to a solid culture medium for culture, and drying and crushing the culture after culture is finished to form the Acremonium culture.
Optionally, the carbon sources in the test tube medium, the shake flask medium and the fermenter medium are: one or more of beef extract, peptone, peanut cake powder, amino acid, gelatin, glucose, sucrose, starch, molasses and whey.
Optionally, the nitrogen sources in the test tube medium, the shake flask medium and the fermenter medium are: one or more of beef extract, yeast extract, soybean powder, cake meal, silkworm chrysalis meal, urea, peptone and gelatin.
Optionally, the carbon source or the nitrogen source of the solid medium is: one or more of corn flour, rice flour, soybean meal, silkworm chrysalis meal, bran, bean hull, straw, rice hull powder, rice and fish meal.
The application has one or more of the following advantages:
the use of the acremonium culture or the acremonium culture of the application for increasing the number of clostridium in the gastrointestinal tract of an animal; the use of a culture of Acremonium terricola in a feed additive or a medicament; use of a culture of acremonium for ameliorating inflammation in the gastrointestinal tract of an animal.
Detailed Description
The following description of the technical solutions in the embodiments of the present disclosure will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present disclosure, but not all embodiments. All other embodiments, which can be made by one of ordinary skill in the art based on the embodiments in this disclosure without inventive faculty, are intended to fall within the scope of this disclosure.
The application discloses an application of a Acremonium terricola culture in improving the content of clostridium prasugrel. The application also discloses application of the Acremonium terricola culture in preparation of medicines, feeds, feed additives or health-care products. Acremonium terricola cultures are effective in increasing the content of Clostridium praecox.
The application discloses a feed additive, which is in a dry powder or liquid form by adding a Acremonium terrestris culture, and the feed additive is a feed additive which exists in any form in a broad sense, but aims at non-treatment and improves the improvement or health care effect; the strengthening method comprises the following steps: adding dry powder of Acremonium terrestris culture in a proportion of 0.001-3% into raw materials or necessary foods, and adding the dry powder into complete feed for use; the feed is added in the feed processing process; adding into the finished product; biologically adding; adding by a physical and chemical method; the adding form is as follows: (1) dry mixing: for example, weighing 1kg of Acremonium terricola culture, adding into 1 ton of complete feed (0.1% of the total feed), uniformly mixing step by step, and directly feeding; (2) Preparing solution by weighing required Acremonium terrestris culture according to the proportion of 0.1%, dissolving in water, stirring uniformly, and adding Acremonium terrestris culture in liquid form, such as: weighing the needed Acremonium terrestris culture according to the proportion of 0.1 percent, and dissolving the Acremonium terrestris culture in water; or other means of addition.
The feed additive containing the Acremonium terrestris culture can improve the content of clostridium prasugrel in the gastrointestinal tract of animals, thereby reducing intestinal inflammation.
Regarding the acquisition of the culture of the Acremonium, in particular, the acquisition is carried out in the form of the culture of the Acremonium or the acquisition is carried out in the form of solid fermentation or liquid fermentation, the Acremonium is purchased in the market, and for convenience of explanation and verification, the illustrated Acremonium is purchased in the institute of China academy of sciences microbiology, in particular, is stored in the institute of China academy of sciences microbiology at the date of 2018 and 4, which is only one purchasing channel of the Acremonium, and of course, the acquisition can also be carried out from other channels; regarding the procurement of Acremonium culture, it can also be procured from the following manufacturers, for example: guangdong big organism stock limited, etc.
Details of the preparation method for Acremonium terricola cultures:
a method for preparing a culture of Acremonium cultivated by liquid cultivation comprises the following steps: fermentation bacteria liquid: acremonium terrestris (Acremonium terricola) is preserved in China academy of sciences microbiological institute at 8.4.2018 with the preservation number of CGMCC 7.357;
test tube strain: the test tube culture medium is as follows: agar 2%, peptone 10%, glucose 20%, and sterilizing at 121deg.C for 20min. Three-point inoculation after cooling, and placing in a biochemical incubator at 25 ℃ for culturing for 5-7 days.
As in the examples, test tube species may be selected: the test tube strain culture medium is as follows: agar 20g/L, peptone 100g/L, glucose 200g/L, and sterilizing at 121deg.C for 20min. Three-point inoculation after cooling, and placing in a biochemical incubator at 25 ℃ for culturing for 5-7 days.
Shaking the bottle strain: the shake flask strain culture medium is as follows: 10% of potato powder, 5% of glucose, 0.5% of dipotassium hydrogen phosphate and 0.5% of magnesium sulfate. Sterilizing at 121deg.C for 20min. Inoculating with test tube strain after cooling, and culturing in shake incubator at 25deg.C for 5-7 days.
As in the examples, shake flask species may be selected: the shake flask strain culture medium is as follows: 100g/L of potato powder, 50g/L of glucose, 5g/L of dipotassium hydrogen phosphate and 5g/L of magnesium sulfate. Sterilizing at 121deg.C for 20min. Inoculating with test tube strain after cooling, and culturing in shake incubator at 25deg.C for 5-7 days.
Fermentation tank strain: the strain culture medium of the fermentation tank is as follows: 10% of potato powder, 5% of glucose, 0.5% of dipotassium hydrogen phosphate, 0.5% of magnesium sulfate and 200L of fermentation tank volume. Sterilizing at 121deg.C for 30min. After cooling, 200ml of shaking flask strains were inoculated, at an ambient temperature of 26℃and aeration culture was carried out for 48 hours during the culture.
As in the examples, fermenter species media may be selected as: 100g/L of potato powder, 50g/L of glucose, 5g/L of dipotassium hydrogen phosphate, 5g/L of magnesium sulfate and 200L of fermentation tank volume. Sterilizing at 121deg.C for 30min. After cooling, 200ml of shaking flask strains were inoculated, at an ambient temperature of 26℃and aeration culture was carried out for 48 hours during the culture.
Liquid mycelium preparation: pumping out fermentation liquor after the fermentation tank is cultured, centrifuging, taking out mycelium at the lower layer, drying at 80-85 ℃, controlling the moisture of the dried mycelium below 8%, and crushing and packaging for later use.
The content of the active ingredients of the Acremonium culture obtained by liquid fermentation of mycelium is shown in Table 1:
TABLE 1 content of active ingredients of Acremonium culture
| Index (I) | Content of |
| Adenosine (mg/kg) | 812 |
| Ergosterol (mg/kg) | 1008 |
| Polysaccharide (%) | 3.4 |
| Uridine (mg/kg) | 987 |
| Guanosine (mg/kg) | 913 |
A method for preparing a culture of Acremonium cultivated by solid cultivation comprises the following steps:
fermentation bacteria liquid: acremonium terricolum (Acremonium terricola) was deposited at the China academy of sciences microbiological study, at 8.4.2018;
test tube strain: the test tube strain culture medium is as follows: agar 2%, peptone 10%, glucose 20%, and sterilizing at 121deg.C for 20min. Three-point inoculation after cooling, and placing in a biochemical incubator at 25 ℃ for culturing for 5-7 days.
As in the examples, test tube species may be selected: the test tube strain culture medium is as follows: agar 20g/L, peptone 100g/L, glucose 200g/L, and sterilizing at 121deg.C for 20min. Three-point inoculation after cooling, and placing in a biochemical incubator at 25 ℃ for culturing for 5-7 days.
Shaking the bottle strain: the shake flask strain culture medium is as follows: 10% of potato powder, 5% of glucose, 0.5% of dipotassium hydrogen phosphate, 0.5% of magnesium sulfate and 20min of sterilization at 121 ℃. Inoculating with test tube strain after cooling, and culturing in shake incubator at 25deg.C for 5-7 days.
As in the examples, shake flask species may be selected: the shake flask strain culture medium is as follows: 100g/L of potato powder, 50g/L of glucose, 5g/L of dipotassium hydrogen phosphate and 5g/L of magnesium sulfate. Sterilizing at 121deg.C for 20min. Inoculating with test tube strain after cooling, and culturing in shake incubator at 25deg.C for 5-7 days.
Fermentation tank strain: the strain culture medium of the fermentation tank is as follows: 10% of potato powder, 5% of glucose, 0.5% of dipotassium hydrogen phosphate, 0.5% of magnesium sulfate and 200L of fermentation tank volume. Sterilizing at 121deg.C for 30min. After cooling, 200ml of shaking flask strains were inoculated, at an ambient temperature of 26℃and aeration culture was carried out for 48 hours during the culture.
As in the examples, fermenter species media may be selected as: 100g/L of potato powder, 50g/L of glucose, 5g/L of dipotassium hydrogen phosphate, 5g/L of magnesium sulfate and 200L of fermentation tank volume. Sterilizing at 121deg.C for 30min. After cooling, 200ml of shaking flask strains were inoculated, at an ambient temperature of 26℃and aeration culture was carried out for 48 hours during the culture.
Inoculating the strain of the fermentation tank into a sterilized and cooled solid culture medium for culture, wherein the inoculation amount is 10%, placing the inoculated material into a woven bag for culture, maintaining the room temperature at 25 ℃, drying the material after 96 hours of culture, controlling the temperature of a hot air circulation oven at 85-90 ℃, controlling the water content of the final material to be below 8%, and crushing and packaging by 60 meshes for later use.
Wherein the solid fermentation culture medium comprises 20% of corn flour, 10% of rice flour, 15% of soybean meal, 30% of silkworm chrysalis meal, 15% of bran, 10% of bean shell, and pulse vacuum sterilization at 121 ℃ for 90min, and cooling after sterilization.
As in the examples, the solid fermentation medium is selected from corn flour 200g/kg, rice flour 100g/kg, soybean meal 150g/kg, silkworm chrysalis meal 300g/kg, bran 150g/kg, soybean hull 100g/kg, and pulse vacuum sterilizing at 121deg.C for 90min, and cooling after sterilization.
The content of the active ingredients of the Acremonium acutum culture obtained by solid fermentation is shown in Table 2:
TABLE 2 content of active ingredients in Acremonium culture
| Index (I) | Content of |
| Adenosine (mg/kg) | 1035 |
| Ergosterol (mg/kg) | 1952 |
| Polysaccharide (%) | 4.5 |
| Uridine (mg/kg) | 1518 |
| Guanosine (mg/kg) | 1078 |
Animal test verification:
examples of use on pigs
And (3) establishing an animal model: pigs, 40, are divided into 4 groups of 10, and a control group and a test group are respectively arranged. Daily feed intake of test group and control group, wherein the test group Acremonium culture is added in the form of feed additive, the addition ratio is 0.001%,0.1%,3%, and other conditions test group and control group are consistent; after 21 days of column feeding, feces are taken and subjected to 16s genome sequencing, and the 16s gene sequencing method is as follows: after the DNA sample is received, the sample is tested; constructing a library by using the sample which is qualified in detection: recovering the target Amplicon fragment, repairing the cohesive end formed by breaking into a flat end by using T4 DNA Polymerase, klenow DNA Polymerase and T4PNK, and adding a base "A" to the 3 'end to ensure that the DNA fragment can be connected with a special connector with a "T" base at the 3' end; or designing and synthesizing a double Index fusion primer containing a sequencing joint, carrying out fusion primer PCR by taking genomic DNA as a template, screening a target Amplicon fragment by using magnetic beads, and finally, carrying out cluster preparation and sequencing by using a qualified library. And (3) carrying out corresponding biological information analysis by using the data obtained by the machine, and obtaining the following results:
the effect of different addition ratios of Acremonium culture on the abundance of the Clostridium praecox flora is shown in Table 3:
TABLE 3 influence of different addition ratios of Acremonium cultures on the abundance of the gastrointestinal Prazimuthing group in pigs
From the above experiments, it can be seen that the Acremonium terrestris culture as a feed additive can indeed increase the content of Clostridium praecox in the gastrointestinal tract of pigs, and the difference is significant.
Examples of use on chickens:
chicken test group and control group: chickens are 16 columns, four columns are a group, 7 chickens are arranged in each column, and a control group and a test group are respectively arranged. Daily feed intake of test group and control group, wherein the test group Acremonium culture is added in the form of feed additive, the addition ratio is 0.001%,0.1%,3%, and other conditions test group and control group are consistent; after 21 days of column feeding, feces are taken and subjected to 16s genome sequencing, and the 16s gene sequencing method is as follows: after the DNA sample is received, the sample is tested; constructing a library by using the sample which is qualified in detection: recovering the target Amplicon fragment, repairing the cohesive end formed by breaking into a flat end by using T4 DNA Polymerase, klenow DNA Polymerase and T4PNK, and adding a base "A" to the 3 'end to ensure that the DNA fragment can be connected with a special connector with a "T" base at the 3' end; or designing and synthesizing a double Index fusion primer containing a sequencing joint, carrying out fusion primer PCR by taking genomic DNA as a template, screening a target Amplicon fragment by using magnetic beads, and finally, carrying out cluster preparation and sequencing by using a qualified library. The corresponding bioinformatic analysis was performed using the data obtained from the off-machine to obtain the following table 4:
TABLE 4 influence of different addition ratios of Acremonium cultures on the abundance of the gastrointestinal Prazimuthally of chickens
From the above experiments, it can be seen that the Acremonium terrestris culture as a feed additive can indeed increase the content of Clostridium praecox in the gastrointestinal tract of chickens, and the difference is significant.
Of course, the above-mentioned culture of Acremonium has only been verified to be capable of increasing the abundance of the group of Clostridium praecox in the gastrointestinal tract of pigs and chickens, and it is not exhaustive to limit that the culture of Acremonium praecox can be used only for the group of Clostridium praecox in the gastrointestinal tract of pigs and chickens, and can be used for cattle, pets, or the like in practical use.
Acremonium species Cordyceps species, which culture can increase the abundance of the group of Prazium bacteria in the gastrointestinal tract of pigs and chickens, other species of Cordyceps species cultures (e.g., cordyceps sinensis, paecilomyces hepiali, cordyceps militaris, cordyceps cicadae, cordyceps guangdongensis, etc.) may also increase the abundance of the group of Prazium bacteria in the gastrointestinal tract of pigs and chickens, and likewise, acremonium species cultures may also increase the abundance of the group of Prazium bacteria in the gastrointestinal tract of other animals.
In the description of the present specification, the descriptions of the terms "one embodiment," "example," "specific example," and the like, mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present disclosure. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiments or examples. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The foregoing has shown and described the basic principles, principal features, and advantages of the present disclosure. It will be understood by those skilled in the art that the present disclosure is not limited to the embodiments described above, and that the embodiments and descriptions described herein are merely illustrative of the principles of the disclosure, and various changes and modifications may be made without departing from the spirit and scope of the disclosure, which are within the scope of the disclosure as claimed.
Claims (2)
1. Use of a culture of acremonium to prepare a feed additive for increasing the number of clostridium prasugrel in the gastrointestinal tract of chickens or pigs;
the Acremonium terricola culture is obtained by a preparation method of liquid fermentation or solid fermentation;
the Acremonium terricola culture comprises the following steps by liquid fermentation:
the method comprises the steps of inoculating Acremonium to a test tube culture medium for culture, inoculating Acremonium after culture in the test tube culture medium to a shake flask culture medium for culture, inoculating Acremonium after culture in the shake flask culture medium to a fermentation tank culture medium for culture, pumping out fermentation liquor after the strain culture in the fermentation tank is finished, centrifuging, drying and crushing to form Acremonium culture;
the Acremonium terricola culture comprises the following steps through solid fermentation:
the method comprises the steps of inoculating Acremonium to a test tube culture medium for culture, inoculating Acremonium after culture in the test tube culture medium to a shake flask culture medium for culture, inoculating Acremonium after culture in the shake flask culture medium to a fermentation tank culture medium for culture, inoculating the fermentation tank strain to a solid culture medium for culture, and drying and crushing the culture after the culture is finished to form Acremonium culture;
the carbon sources in the test tube strain culture medium, the shake flask strain culture medium and the fermentation tank strain culture medium are as follows: one or more of beef extract, peptone, peanut cake powder, amino acid, gelatin, glucose, sucrose, starch, molasses and whey;
the nitrogen sources in the test tube strain culture medium, the shake flask strain culture medium and the fermentation tank strain culture medium are as follows: one or more of beef extract, yeast extract, soybean powder, cake meal, silkworm chrysalis meal, urea, peptone and gelatin;
the carbon source or the nitrogen source of the solid culture medium is as follows: one or more of corn flour, rice flour, soybean meal, silkworm chrysalis meal, bran, bean hull, straw, rice hull powder, rice and fish meal.
2. The use according to claim 1, characterized in that the culture of acremonium terrestris contains the following active substances in the following amounts: 0.001-5% of adenosine, 0.002-4% of ergosterol, 0.002-4% of guanosine, 0.002-4% of uridine and 0.002-10% of cordyceps polysaccharide.
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| CN103734482A (en) * | 2014-01-20 | 2014-04-23 | 安徽康智生物科技有限责任公司 | Production method of feed additive 'Acremonium terricola culture' |
| CN107772076A (en) * | 2016-08-31 | 2018-03-09 | 辽宁弘侨生物科技股份有限公司 | A kind of immune potentiator for wormgrass base feed and preparation method thereof |
| CN109055241A (en) * | 2018-08-30 | 2018-12-21 | 广东容大生物股份有限公司 | A method of preparing mortierella Diding culture |
| CN112538436A (en) * | 2020-12-23 | 2021-03-23 | 江苏三仪生物工程有限公司 | Preparation method of acremonium terricola culture |
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| CN103734482A (en) * | 2014-01-20 | 2014-04-23 | 安徽康智生物科技有限责任公司 | Production method of feed additive 'Acremonium terricola culture' |
| CN107772076A (en) * | 2016-08-31 | 2018-03-09 | 辽宁弘侨生物科技股份有限公司 | A kind of immune potentiator for wormgrass base feed and preparation method thereof |
| CN109055241A (en) * | 2018-08-30 | 2018-12-21 | 广东容大生物股份有限公司 | A method of preparing mortierella Diding culture |
| CN112538436A (en) * | 2020-12-23 | 2021-03-23 | 江苏三仪生物工程有限公司 | Preparation method of acremonium terricola culture |
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