CN116144536A - Composite probiotics fermentation composition for improving male sperm motility and application - Google Patents
Composite probiotics fermentation composition for improving male sperm motility and application Download PDFInfo
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Abstract
The invention belongs to the technical field of traditional Chinese medicine fermentation, and particularly relates to a composite probiotic fermentation composition for improving male sperm motility and application thereof. The composition comprises the following components: bifidobacterium lactis, bifidobacterium bifidum, bifidobacterium animalis, bifidobacterium longum, lactobacillus salivarius, lactobacillus casei, lactobacillus helveticus, lactobacillus paracasei, lactobacillus plantarum, lactobacillus reuteri, lactobacillus rhamnosus, lactobacillus acidophilus; the bifidobacterium animalis is a mixture of bifidobacterium animalis BB-12, bifidobacterium animalis B94 and bifidobacterium animalis ZK-77 with the mass ratio of 1:1-3:6-10. The composite bacteria can be used for fermenting the traditional Chinese medicine composition to obtain fermentation liquor, so that the activity of male sperms can be effectively improved, the prepared traditional Chinese medicine liquid can be directly used for preparing downstream products for improving the activity of male sperms, and the traditional Chinese medicine liquid can also be prepared into a remanufacturing agent after drying, and the preparation method is simple and environment-friendly.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicine fermentation, and particularly relates to a composite probiotic fermentation composition for improving male sperm motility and application thereof.
Background
The causes of male infertility include sexual dysfunction, varicocele, genital tract infection, congenital dysplasia, acquired sexual disease, endocrine disturbance, and immunity factor. However, most of them are male infertility caused by unknown causes, which are only manifested by sperm quality abnormalities such as oligospermia, oligospermia and/or teratospermia, and the male infertility caused by unknown causes may be caused by various factors such as endocrine disruption, active oxygen and gene defects caused by long-term stress environmental factors.
The current western medicine treatment mainly comprises: (1) artificial insemination: is the most effective treatment method for male infertility, and uses various physical and biochemical techniques to treat semen to improve the conception ability of sperm; (2) treatment of genital tract inflammation: a series of male infertility treatment methods using antibiotics in combination with anti-inflammatory agents are generally claimed; (3) immunotherapy: surgical operation is applied to cut off local lesion focus of the genital tract, so that the generation of anti-sperm antibody is reduced, and an immune preparation is used at the same time; (4) endocrine treatment: the long-acting testosterone enanthate is mainly used for treating hypogonadism of gonadotrophin. But western medicines have great side effects on the body and sperm quality of patients.
The traditional Chinese medicine has unique advantages in improving the sperm quality of men, and improves the fatigue resistance of organisms, the sperm motility and the sperm quality and the hormone level through overall regulation.
The Chinese patent application CN108853469A discloses a composition for improving male sperm motility and male erectile function and a preparation process thereof, wherein the composition is prepared from deer products such as deer meat, deer tendons, deer tails, deer penis, deer kidneys and the like, marine fish oligopeptide powder, maca powder, rhizoma polygonati, chinese yam, raspberry, fructus alpiniae oxyphyllae and the like, and particularly is prepared from auxiliary materials such as xylitol and the like in a form of pressed candy. The composition can be used as health products, health foods, health beverages or medical products. The composition of the invention has excellent effects of enhancing kidney qi of human body and improving kidney function. In addition, the composition of the invention has excellent effect of improving male sperm motility and male erectile function.
Another chinese patent application CN103768139a discloses a medicament for improving sperm quantity and activity of male, and its preparation, application and preparation method, wherein the medicament and its preparation are prepared from hickory nut kernel, siraitia grosvenorii, zinc gluconate and mannans, and the above-mentioned medicaments are added with auxiliary materials, and made into medicinal preparation according to conventional preparation process. The medicament and the preparation thereof have unexpected treatment effects when applied to enhancing the number and the vitality of male sperms and improving the sperm quality, and are another option for treating male infertility.
Also, chinese patent application CN107913357A discloses a new use of a traditional Chinese medicine composition in preparing a medicine/health care product for treating low sperm motility, wherein the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 5 to 30 parts of cordyceps cephalosporin powder, 1 to 20 parts of mushroom, 1 to 20 parts of lucid ganoderma, 1 to 20 parts of dogwood, 5 to 30 parts of cistanche, 5 to 30 parts of fragrant solomonseal rhizome and 1 to 20 parts of dwarf lilyturf tuber. The composition can improve various indexes of sperm motility, has small toxic and side effects and has good clinical application prospect.
Although the above patents all have studied to improve male fertility and sperm motility and obtain preliminary results, the amounts of rare medicinal materials such as ganoderma lucidum, hairy antler are large, and the preparation and the like cannot fully utilize the active ingredients of the medicinal materials, so that the efficacy still needs to be further improved.
The fermented Chinese medicine is a novel medicine which takes one or more probiotics in optimized intestinal probiotics flora as a strain, utilizes a microecology and bionics method to simulate the intestinal environment of a human body and the digestion and decomposition process of Chinese medicine components in the human body in vitro by using a biological grafting method, carries out biological conversion on the extracted Chinese medicine active components, converts macromolecular substances of the Chinese medicine into small molecular components which can be directly absorbed by the intestinal tract of the human body through microorganism, and ensures that the Chinese medicine is quickly absorbed and quantitatively treated.
Disclosure of Invention
In order to overcome the technical problems, the invention provides the composite probiotic fermentation composition for improving the sperm motility of men, which has the advantages of less medicinal material consumption, high utilization rate, definite medicinal effect for improving the sperm motility and good social value.
In order to achieve the above object, the technical scheme provided by the invention is as follows:
the bifidobacterium animalis ZK-77 is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.23681 and the preservation date of 2021, 10 months and 28 days.
The bifidobacterium animalis (Bifidobacterium animalis) provided by the invention is classified and named as bifidobacterium animalis Bifidobacterium animalis; the preservation unit is China general microbiological culture Collection center, the preservation number is CGMCC No.23681, and the preservation address is: beijing, chaoyang area, north Chen Xili No. 1, 3, china academy of sciences, microbiological institute.
A composite probiotic fermented composition, the composition comprising the following components: bifidobacterium lactis, bifidobacterium bifidum, bifidobacterium animalis, bifidobacterium longum, lactobacillus salivarius, lactobacillus casei, lactobacillus helveticus, lactobacillus paracasei, lactobacillus plantarum, lactobacillus reuteri, lactobacillus rhamnosus, lactobacillus acidophilus;
preferably, the bifidobacterium animalis is a mixture of bifidobacterium animalis BB-12, bifidobacterium animalis B94 and bifidobacterium animalis ZK-77 in a mass ratio of 1:1-3:6-10.
Preferably, the bifidobacterium lactis is a mixture of bifidobacterium lactis HN019 and bifidobacterium lactis Bi07 in a mass ratio of 1:2-5.
Preferably, the lactobacillus salivarius is lactobacillus salivarius ZK-88, which is preserved in China general microbiological culture Collection center with a preservation number of CGMCC No.23682 and a preservation date of 2021, 10 months and 28 days.
The lactobacillus salivarius (Lactobacillus salivarius) provided by the invention is classified and named as lactobacillus salivarius Lactobacillus salivarius; the preservation address is: beijing, chaoyang area, north Chen Xili No. 1, 3, china academy of sciences, microbiological institute.
Preferably, the composition comprises the following components: bifidobacterium lactis HN019, bifidobacterium bifidum Bb06, bifidobacterium animalis Bb-12, bifidobacterium lactis Bi07, bifidobacterium longum R0175, bifidobacterium animalis B94, lactobacillus salivarius ZK-88, lactobacillus casei LC11, lactobacillus helveticus R0052, lactobacillus paracasei Lpc37, lactobacillus plantarum R1012, lactobacillus reuteri HA188, lactobacillus rhamnosus R0011, lactobacillus acidophilus NCFM, and bifidobacterium animalis ZK-77.
Preferably, the composition comprises the following components in parts by weight: 6-7 parts of bifidobacterium lactis, 1-3 parts of bifidobacterium bifidum, 10-20 parts of bifidobacterium animalis, 5-10 parts of bifidobacterium longum, 10-20 parts of lactobacillus salivarius, 2-12 parts of lactobacillus casei, 5-10 parts of lactobacillus helveticus, 6-10 parts of lactobacillus paracasei, 15-20 parts of lactobacillus plantarum, 1-6 parts of lactobacillus reuteri, 3-9 parts of lactobacillus rhamnosus and 5-12 parts of lactobacillus acidophilus.
It is still another object of the present invention to provide a composite probiotic fermented composition, the preparation method of which comprises the steps of:
(1) Pulverizing the Chinese medicinal composition, dispersing in water, performing enzymolysis, and inactivating enzyme to obtain enzymolysis system; the traditional Chinese medicine comprises the following components: astragalus root, red ginseng, epimedium herb, sealwort, schisandra fruit and dodder seed;
(2) Adding a culture medium accounting for 2% of the mass of the enzymolysis system, inoculating the composite probiotic composition, fermenting, and centrifuging to obtain a centrifugate and a precipitate;
(3) Extracting the precipitate with ethanol water solution to obtain ethanol extract;
(4) Mixing the centrifugate in step (2) with ethanol extract, and freeze drying to obtain the final product.
Preferably, the traditional Chinese medicine in the step (1) comprises the following components: 5-10 parts of astragalus membranaceus, 1-5 parts of red ginseng, 1-5 parts of epimedium herb, 1-5 parts of rhizoma polygonati, 1-5 parts of schisandra chinensis and 1-5 parts of semen cuscutae;
preferably, the enzymolysis is performed with a complex enzyme, wherein the complex enzyme is 1:1-2 of cellulase and papain, the mass of the complex enzyme being 1-5% of the volume of water;
preferably, the enzymolysis temperature is 55-60 ℃, the enzymolysis pH is 5.0-7.5, and the enzymolysis time is 2-4h.
Preferably, the composition of the medium in step (2) is: 10-15% of soybean powder, 8-14% of sucrose, 6-12% of peptone, 0.1-0.2% of potassium dihydrogen phosphate, 0.1-0.5% of magnesium sulfate and the balance of water; fermenting and culturing for 60-72h at 35-40 ℃;
preferably, the ethanol water solution in the step (3) is 65-75% ethanol water solution, and the extraction is heating reflux extraction, and the extraction time is 0.5-2h.
It is also an object of the present invention to provide the use of the above fermentation composition for the preparation of a product for improving sperm motility in men.
The invention also aims to provide a separation method of the bifidobacterium animalis ZK-77 strain lactobacillus salivarius ZK-88, which comprises the following steps: is obtained by separating from human mouth and screening at 37-42 deg.C.
Preferably, the separation method comprises the steps of: 100ul of human mouth rinse solution sample is taken and diluted in a gradient of 10 -1 -10 -4 Double, respectively coating on a selective culture medium flat plate of MRS+bromocresol purple and MRS+mupirocin lithium salt, placing at 37 ℃, culturing overnight under anaerobic condition, picking single colony for purification, and identifying and preserving seeds for 16 seconds;
16S identification: using universal primers 27F (AGAGTTTGATCCTGGCTCAG) and 1492R
(TACGGCTACCTTGTT-ACGACTT) to generate a 16S PCR fragment, and submitting it to the sequencing (forward) by the department of Optimago.
Compared with the prior art, the invention has the technical advantages that:
(1) The invention provides a new strain, provides a microbial inoculum containing the strain, and prepares a composite probiotic composition, which can be used for fermenting traditional Chinese medicine liquid, has better synergistic effect among strains, can effectively improve the fermentation effect of the probiotic composition on traditional Chinese medicine materials, and improves the drug effect of the traditional Chinese medicine materials.
(3) The traditional Chinese medicine fermentation composition provided by the invention can effectively improve the male sperm motility, can be directly used for preparing products for improving the male sperm motility, can also be used for preparing remanufacturing agents after drying, and is simple and green in preparation method.
(4) The product obtained in the invention contains specific composite probiotics and a fermented composition thereof, and the specific composite probiotics and the fermented composition cooperate to promote and improve the activity of male sperms.
Detailed Description
The present invention will be described by way of specific examples, to facilitate understanding and grasping of the technical solution of the present invention, but the present invention is not limited thereto. The experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagents and materials, unless otherwise specified, are commercially available; and the different sources have no significant effect on the product performance.
The probiotics in the following examples are all commercial conventional raw materials. Wherein suppliers of bifidobacterium lactis HN019, bifidobacterium lactis Bi07, lactobacillus paracasei Lpc37, lactobacillus acidophilus NCFM, lactobacillus casei LC11 and bifidobacterium bifidum Bb06 are DuPont Danesk groups; the supplier of Bifidobacterium animalis BB-12 is Hansen, denmark; the suppliers of lactobacillus helveticus R0052, lactobacillus plantarum R1012, bifidobacterium animalis B94, bifidobacterium longum R0175, lactobacillus rhamnosus R0011, lactobacillus reuteri HA188 are canadian raman groups.
EXAMPLE 1 isolation and characterization of bifidobacterium animalis ZK-77 and Lactobacillus salivarius ZK-88
100ul of human mouth rinse solution sample is taken and diluted in a gradient of 10 -1 -10 -4 Double, respectively coating on MRS+bromocresol purple and MRS+mupirocin lithium salt selective culture medium plates, placing at 37 ℃ for anaerobic overnight culture, respectively picking single colony for purification, adopting universal primers 27F (AGAGTTTGATCCTGGCTCAG) and 1492R (TACGGCTACCTTGTT-ACGACTT) to generate 16S PCR fragments, submitting the fragments to the sequencing (forward direction) of the Optimago company, and adopting 16S to identify that the colonies picked in the two culture mediums are lactobacillus salivarius (MRS+bromocresol purple culture medium) and bifidobacterium animalis (MRS+mupirocin lithium salt culture medium) respectively; 16S identification: the 16S PCR fragment was generated using the universal primers 27F (AGAGTTTGATCCTGGCTCAG) and 1492R (TACGGCTACCTTGTT-ACGACTT), submitted to the department of Prinsepia sequencing (forward), and passed.
The bifidobacterium animalis (Bifidobacterium animalis) provided by the invention is classified and named as bifidobacterium animalis Bifidobacterium animalis, and is preserved in the China general microbiological culture collection center (CGMCC) with the preservation number of CGMCC NO.23681; the preservation unit is China general microbiological culture Collection center, and the preservation address is: the collection date of the institute of microbiology of the national academy of sciences of China is 2021, 10 months and 28 days.
The lactobacillus salivarius (Lactobacillus salivarius) provided by the invention is classified and named as lactobacillus salivarius Lactobacillus salivarius; the preservation unit is China general microbiological culture Collection center, the preservation number is CGMCC No.23682, and the preservation address is: the collection date of the institute of microbiology of the national academy of sciences of China is 2021, 10 months and 28 days.
Example 2
The composite probiotic composition comprises the following components in parts by weight: 6 parts of bifidobacterium lactis, 06 parts of bifidobacterium bifidum Bb06, 10 parts of bifidobacterium animalis, R0175 parts of bifidobacterium longum, ZK-88 parts of lactobacillus salivarius, LC115 parts of lactobacillus casei, R0052 parts of lactobacillus helveticus, lpc37 parts of lactobacillus paracasei, R1012 15 parts of lactobacillus plantarum, HA188 1 parts of lactobacillus reuteri, R0011 parts of lactobacillus rhamnosus and NCFM 5 parts of lactobacillus acidophilus.
Wherein the bifidobacterium lactis is a mixture of bifidobacterium lactis HN019 and bifidobacterium lactis Bi07 in a mass ratio of 1:2; the bifidobacterium animalis is a mixture of bifidobacterium animalis BB-12, bifidobacterium animalis B94 and bifidobacterium animalis ZK-77 in a mass ratio of 1:3:6.
The viable bacteria specification of the composite probiotic composition is 220 hundred million CFU/g.
Example 3
The composite probiotic composition of the example 15 comprises the following components in parts by weight: 7 parts of bifidobacterium lactis, 06 parts of bifidobacterium bifidum Bb06, 20 parts of bifidobacterium animalis, 0175 10 parts of bifidobacterium longum R, 20 parts of lactobacillus salivarius ZK-88, 11 parts of lactobacillus casei LC11, 0052 10 parts of lactobacillus helveticus R, 37 parts of lactobacillus paracasei Lpc, 1012 20 parts of lactobacillus plantarum R1012 20 parts, 188 parts of lactobacillus reuteri HA, 0011 parts of lactobacillus rhamnosus R0011 and 12 parts of lactobacillus acidophilus NCFM.
Wherein the bifidobacterium lactis is a mixture of bifidobacterium lactis HN019 and bifidobacterium lactis Bi07 in a mass ratio of 1:4; the bifidobacterium animalis is a mixture of bifidobacterium animalis BB-12, bifidobacterium animalis B94 and bifidobacterium animalis ZK-77 in a mass ratio of 1:1:8.
The viable bacteria specification of the composite probiotic composition is 220 hundred million CFU/g.
Example 4
The preparation method of the fermentation composition is characterized by comprising the following steps:
(1) Crushing 5 parts of astragalus, 1 part of red ginseng, 1 part of epimedium herb, 1 part of rhizoma polygonati, 1 part of shizandra berry and 1 part of semen cuscutae, dispersing in 8 times of water, adding 2% of water quality complex enzyme, carrying out enzymolysis for 2 hours at the temperature of 55 ℃ and the pH value of 5.0, and inactivating the enzyme at the temperature of 100 ℃ for 5 minutes to prepare an enzymolysis system; the complex enzyme is 1:1 and papain;
(2) Adding a culture medium with the mass of 2% into an enzymolysis system, inoculating a composite probiotic composition of example 2 with the weight of 0.5% of the enzymolysis system, fermenting and culturing for 72 hours at 35 ℃, and centrifuging to obtain a centrifugate and a precipitate; wherein the culture medium comprises 10% of soybean powder, 14% of sucrose, 6% of peptone, 0.1% of potassium dihydrogen phosphate, 0.1% of magnesium sulfate and the balance of water;
(3) Reflux-extracting the precipitate with 5 times of 65% ethanol water solution for 0.5 hr to obtain ethanol extractive solution;
(4) Mixing the centrifugate in (2) with the ethanol extract in (3), and freeze-drying to obtain fermentation product.
Example 5
The preparation method of the fermentation composition is characterized by comprising the following steps:
(1) Crushing 10 parts of astragalus, 3 parts of red ginseng, 3 parts of epimedium herb, 3 parts of rhizoma polygonati, 3 parts of shizandra berry and 3 parts of semen cuscutae, dispersing in 8 times of water, adding 2% of water compound enzyme, carrying out enzymolysis for 3h at 55 ℃ and pH of 6.0, and inactivating enzyme at 100 ℃ for 5min to obtain an enzymolysis system; the complex enzyme is 1:1 and papain;
(2) Adding a 2% culture medium into an enzymolysis system, inoculating 0.5% of the composite probiotic composition of the example 2, fermenting and culturing at 35 ℃ for 72 hours, and centrifuging to obtain a centrifugate and a precipitate; wherein the culture medium comprises 15% of soybean powder, 8% of sucrose, 6% of peptone, 0.2% of potassium dihydrogen phosphate, 0.5% of magnesium sulfate and the balance of water;
(3) Reflux-extracting the precipitate with 5 times of 75% ethanol water solution for 1 hr to obtain ethanol extractive solution;
(4) Mixing the centrifugate in (2) with the ethanol extract in (3), and freeze drying to obtain fermentation product.
Example 6
The preparation method of the fermentation composition is characterized by comprising the following steps:
(1) Crushing 8 parts of astragalus, 2 parts of red ginseng, 2 parts of epimedium herb, 2 parts of rhizoma polygonati, 2 parts of shizandra berry and 2 parts of semen cuscutae, dispersing in 8 times of water, adding 2% water compound enzyme, carrying out enzymolysis for 4 hours at 55 ℃ and pH5.0, and inactivating enzyme at 100 ℃ for 5 minutes to obtain an enzymolysis system; the complex enzyme is 1:2 and papain;
(2) Adding a culture medium with the mass of 2% into an enzymolysis system, inoculating the composite probiotic composition of the example 3 with the weight of 0.5% of the enzymolysis system, fermenting and culturing for 72 hours at 40 ℃, and centrifuging to obtain a centrifugate and a precipitate; wherein the culture medium comprises 15% of soybean powder, 14% of sucrose, 6% of peptone, 0.1% of potassium dihydrogen phosphate, 0.1% of magnesium sulfate and the balance of water;
(3) Reflux-extracting the precipitate with 5 times of 75% ethanol water solution for 1 hr to obtain ethanol extractive solution;
(4) Mixing the centrifugate in (2) with the ethanol extract in (3), and freeze-drying to obtain fermentation product.
The crude drug amounts of the Chinese medicinal compositions used in examples 4 to 6 were consistent.
Example 7
The difference between this example and example 4 is that the composite probiotic material is different, the specific composition is: 6 parts of bifidobacterium lactis, 06 parts of bifidobacterium bifidum Bb06, 4 parts of bifidobacterium animalis, R0175 parts of bifidobacterium longum, 16 parts of lactobacillus salivarius ZK-88, 11 parts of lactobacillus casei LC, 2 parts of lactobacillus helveticus R0055 parts of lactobacillus paracasei Lpc37 parts of lactobacillus plantarum, R1012 15 parts of lactobacillus reuteri HA188 1 parts of lactobacillus rhamnosus R0011 parts of lactobacillus acidophilus NCFM 5 parts of lactobacillus helveticus.
Wherein the bifidobacterium lactis is a mixture of bifidobacterium lactis HN019 and bifidobacterium lactis Bi07 in a mass ratio of 1:2; the bifidobacterium animalis is a mixture of bifidobacterium animalis BB-12 and bifidobacterium animalis B94 in a mass ratio of 1:3.
The viable bacteria specification of the composite probiotic composition is 220 hundred million CFU/g.
The preparation method is the same as in example 4.
Example 8
The difference from example 4 is that the complex probiotics do not contain lactobacillus salivarius ZK-88; the use amount of the extract is added into bifidobacterium animalis ZK-77, and the specific composition is as follows: the composite probiotic composition comprises the following specific components: the weight portions are as follows: 6 parts of bifidobacterium lactis, 06 parts of bifidobacterium bifidum Bb06, 20 parts of bifidobacterium animalis, R0175 parts of bifidobacterium longum, 115 parts of lactobacillus casei LC, R0052 parts of lactobacillus helveticus, 37 parts of lactobacillus paracasei Lpc, 1012 15 parts of lactobacillus plantarum, HA188 1 parts of lactobacillus reuteri, R0011 parts of lactobacillus rhamnosus and NCFM 5 parts of lactobacillus acidophilus.
Wherein the bifidobacterium lactis is a mixture of bifidobacterium lactis HN019 and bifidobacterium lactis Bi07 in a mass ratio of 1:2; the mass ratio of the bifidobacterium animalis is 1:3:16, bifidobacterium animalis BB-12, bifidobacterium animalis B94, and bifidobacterium animalis ZK-77.
The viable bacteria specification of the composite probiotic composition is 220 hundred million CFU/g.
The preparation method is the same as in example 4.
Example 9
The difference between this pair of examples and example 4 is the fermentation process; in particular to a traditional Chinese medicine composition. The specific process is as follows:
(1) Crushing 5 parts of astragalus, 1 part of codonopsis pilosula, 1 part of morinda officinalis, 1 part of rhizoma polygonati, 1 part of schisandra chinensis and 1 part of cistanche deserticola, dispersing in 8 times of water, adding 2% of water quality complex enzyme, carrying out enzymolysis for 2 hours at the temperature of 55 ℃ and the pH value of 5.0, and inactivating the enzyme at the temperature of 100 ℃ for 5 minutes to obtain an enzymolysis system; the complex enzyme is 1:1 and papain;
(2) Adding a 2% culture medium into an enzymolysis system, inoculating 0.5% of the composite probiotic composition of example 2, fermenting and culturing at 35 ℃ for 72 hours, and centrifuging to obtain a centrifugate and a precipitate; wherein the culture medium comprises 10% of soybean powder, 14% of sucrose, 6% of peptone, 0.1% of potassium dihydrogen phosphate, 0.1% of magnesium sulfate and the balance of water;
(3) Reflux-extracting the precipitate with 5 times of 65% ethanol water solution for 0.5 hr to obtain ethanol extractive solution;
(4) Mixing the centrifugate in (2) with the ethanol extract in (3), and freeze drying to obtain the final product.
Pharmacodynamic experiments
Experimental grouping: SD male rats 8 weeks old, group: blank, model, examples 4,5,6,7,8, 9.
The experimental method comprises the following steps: male mice except the blank group were anesthetized, sterilized, right testis removed, and administration was started 3 days after surgery. Sampling for 4 weeks, taking blood from abdominal aorta after the experiment is finished, measuring the content of serum testosterone, killing rats, weighing prepuce gland, seminal vesicle gland and prostate gland, and calculating viscera coefficient mg/g; the left epididymis was taken, cut vertically 2 times, rinsed with 1mL of physiological saline and incubated for 10min, and then 1 drop was placed in a hemocytometer to count the number of sperm (mL). Another 1 drop was taken, 200 sperm were microscopically observed, the number of motile sperm was recorded, and the sperm motility (sperm motility = motile sperm count/(motile sperm count + motile sperm count) 100%) was calculated.
The experimental results are shown in tables 1-2 below.
TABLE 1 influence on the glandular index of castrated mice
Note that: the model set is compared with the blank set, * p is less than 0.05; in contrast to the set of models, △ p < 0.05, compared with example 4, # P＜0.05。
TABLE 2 analysis of results of sperm count and sperm motility in rats
Note that: the model set is compared with the blank set, * p is less than 0.05; in contrast to the set of models, △ p < 0.05, compared with example 4, # P＜0.05。
the foregoing detailed description is directed to one of the possible embodiments of the present invention, which is not intended to limit the scope of the invention, but is to be accorded the full scope of all such equivalents and modifications so as not to depart from the scope of the invention.
Claims (10)
1. A composite probiotic composition, characterized in that it comprises the following components: bifidobacterium lactis, bifidobacterium bifidum, bifidobacterium animalis, bifidobacterium longum, lactobacillus salivarius, lactobacillus casei, lactobacillus helveticus, lactobacillus paracasei, lactobacillus plantarum, lactobacillus reuteri, lactobacillus rhamnosus, lactobacillus acidophilus; the bifidobacterium animalis is a mixture of bifidobacterium animalis BB-12, bifidobacterium animalis B94 and bifidobacterium animalis ZK-77 with the mass ratio of 1:1-3:6-10.
2. The probiotic composition according to claim 1, wherein said bifidobacterium animalis ZK-77 is deposited at the China general microbiological culture Collection center with a collection number of CGMCC No.23681 and a collection date of 2021, 10 months and 28 days.
3. The probiotic composition according to claim 1, characterized in that the bifidobacterium lactis is a mixture of bifidobacterium lactis HN019, bifidobacterium lactis Bi07 in a mass ratio of 1:2-5.
4. The probiotic composition according to claim 1, wherein the lactobacillus salivarius is lactobacillus salivarius ZK-88 deposited at the China general microbiological culture Collection center with the accession number CGMCC No.23682 and the date of deposit is 2021, 10 months and 28 days.
5. The probiotic composition according to claim 1, characterized in that it comprises the following components: bifidobacterium lactis HN019, bifidobacterium bifidum Bb06, bifidobacterium animalis Bb-12, bifidobacterium lactis Bi07, bifidobacterium longum R0175, bifidobacterium animalis B94, lactobacillus salivarius ZK-88, lactobacillus casei LC11, lactobacillus helveticus R0052, lactobacillus paracasei Lpc37, lactobacillus plantarum R1012, lactobacillus reuteri HA188, lactobacillus rhamnosus R0011, lactobacillus acidophilus NCFM, and bifidobacterium animalis ZK-77.
6. The probiotic composition according to claim 1, characterized in that it comprises the following components in parts by weight: 6-7 parts of bifidobacterium lactis, 1-3 parts of bifidobacterium bifidum, 10-20 parts of bifidobacterium animalis, 5-10 parts of bifidobacterium longum, 10-20 parts of lactobacillus salivarius, 2-12 parts of lactobacillus casei, 5-10 parts of lactobacillus helveticus, 6-10 parts of lactobacillus paracasei, 15-20 parts of lactobacillus plantarum, 1-6 parts of lactobacillus reuteri, 3-9 parts of lactobacillus rhamnosus and 5-12 parts of lactobacillus acidophilus.
7. A composite probiotic fermented composition, characterized in that the preparation method of the composition comprises the following steps:
(1) Pulverizing the Chinese medicinal composition, dispersing in water, performing enzymolysis, and inactivating enzyme to obtain enzymolysis system; the traditional Chinese medicine comprises the following components: astragalus root, red ginseng, epimedium herb, sealwort, schisandra fruit and dodder seed;
(2) Adding a culture medium into the enzymolysis system, inoculating the probiotic composition according to any one of claims 1-6, fermenting, centrifuging to obtain a centrifugate and a precipitate;
(3) Extracting the precipitate with ethanol water solution to obtain ethanol extract;
(4) Mixing the centrifugate of step (2) with ethanol extract, and lyophilizing to obtain fermented composition.
8. The fermentation composition of claim 7, wherein the traditional Chinese medicine in step (1) comprises the following components: 5-10 parts of astragalus membranaceus, 1-5 parts of red ginseng, 1-5 parts of epimedium herb, 1-5 parts of rhizoma polygonati, 1-5 parts of schisandra chinensis and 1-5 parts of semen cuscutae; the enzymolysis is carried out by using complex enzyme, wherein the complex enzyme is 1:1-2 of cellulase and papain, the mass of the complex enzyme being 1-5% of the volume of water; the enzymolysis temperature is 55-60 ℃, the enzymolysis pH is 5.0-7.5, and the enzymolysis time is 2-4h.
9. The fermentation composition of claim 7, wherein the medium in step (2) has a composition of: 10-15% of soybean powder, 8-14% of sucrose, 6-12% of peptone, 0.1-0.2% of potassium dihydrogen phosphate, 0.1-0.5% of magnesium sulfate and the balance of water; fermenting and culturing for 60-72h at 35-40 ℃; the ethanol water solution in the step (3) is 65-75% ethanol water solution, and the extraction is heating reflux extraction, and the extraction time is 0.5-2h.
10. Use of a complex probiotic fermented composition according to any one of claims 7 to 9 for the preparation of a product for improving sperm motility in men.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117286080A (en) * | 2023-11-23 | 2023-12-26 | 吉林农业大学 | Probiotics for improving sperm motility and application thereof |
| CN117701456A (en) * | 2023-12-22 | 2024-03-15 | 善恩康生物科技（苏州）有限公司 | Bifidobacterium lactis, preparation thereof and application thereof in reproductive injury or disease |
| CN117778275A (en) * | 2024-02-23 | 2024-03-29 | 微康益生菌(苏州)股份有限公司 | Probiotic agent for improving oligospermia and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117286080A (en) * | 2023-11-23 | 2023-12-26 | 吉林农业大学 | Probiotics for improving sperm motility and application thereof |
| CN117286080B (en) * | 2023-11-23 | 2024-02-06 | 吉林农业大学 | Probiotics for improving sperm motility and application thereof |
| CN117701456A (en) * | 2023-12-22 | 2024-03-15 | 善恩康生物科技（苏州）有限公司 | Bifidobacterium lactis, preparation thereof and application thereof in reproductive injury or disease |
| CN117778275A (en) * | 2024-02-23 | 2024-03-29 | 微康益生菌(苏州)股份有限公司 | Probiotic agent for improving oligospermia and application thereof |
| CN117778275B (en) * | 2024-02-23 | 2024-06-04 | 微康益生菌(苏州)股份有限公司 | Probiotic agent for improving oligospermia and application thereof |
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