CN115478029B - Lactobacillus reuteri LRB5, microbial inoculum and application - Google Patents

Lactobacillus reuteri LRB5, microbial inoculum and application Download PDF

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CN115478029B
CN115478029B CN202211154794.8A CN202211154794A CN115478029B CN 115478029 B CN115478029 B CN 115478029B CN 202211154794 A CN202211154794 A CN 202211154794A CN 115478029 B CN115478029 B CN 115478029B
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lactobacillus reuteri
lrb5
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CN115478029A (en
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赵勇
钟儒清
陈亮
张宏福
姜岳
张鹏飞
周业勋
郝亚男
卢东欣
房涵涵
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Institute of Animal Science of CAAS
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A61P15/08—Drugs for genital or sexual disorders; Contraceptives for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus

Abstract

The application relates to the field of microorganisms, in particular to lactobacillus reuteri LRB5, a microbial inoculum and application thereof. The lactobacillus reuteri LRB5 can be planted in an animal body, and has strong tolerance to artificial bile salts, artificial gastric acid and artificial intestinal juice; can improve intestinal health, improve intestinal immunity, and increase sperm motility, concentration, pregnancy rate and number of live animals; the strain can improve animal metabolism, increase beneficial metabolite, thereby increasing testis beneficial metabolite such as retinoic acid, and improving sperm quality and fertility of male animals.

Description

Lactobacillus reuteri LRB5, microbial inoculum and application
Technical Field
The application relates to the field of microorganisms, in particular to lactobacillus reuteri LRB5, a microbial inoculum and application thereof.
Background
At present, the male sterility and sterility rate of male animals are greatly improved mainly because the male reproductive system is more easily affected by environmental factors. However, there is no good method to improve semen quality to treat male infertility. In recent years, researches show that intestinal flora is closely related to the health of people and other animals, however, the relation between the intestinal flora and a male reproductive axis is not confirmed, and although researches show that intestinal flora mixtures can change the semen quality of male animals and beneficial intestinal beneficial flora can improve the semen quality of animals, no specific beneficial bacteria can improve the semen quality of animals and improve the fertility of male animals, and no literature discloses specific intestinal bacteria can improve the fertility of male animals. The existing lactobacillus reuteri ATCC PTA 6475 can mainly increase the number of animal testis mesenchyme, increase the content of testosterone and also improve the concentration and activity of sperms. But does not indicate that it can increase the acrosome integrity of sperm, reduce the rate of sperm malformation, and increase fertility, pregnancy rate and number of live animals. While improving the sperm motility and sperm concentration of an animal does not represent a certain ability to improve the fertility of an animal. Many factors affect animal fertility including sperm concentration, sperm motility, sperm acrosome integrity, sperm deformity rate, sperm fertilization capacity, and some physiological functions of the animal body (such as physical ability, etc.). Previous studies by applicant have found that chestnut polysaccharides can increase sperm motility and sperm concentration in animals but cannot increase fertility in animals. Many studies have found similar results.
Disclosure of Invention
The application aims to provide lactobacillus reuteri @Lactobacillus reuteri)B5。
Another object of the present application is to provide a composition comprising the above Lactobacillus reuteriLactobacillus reuteri) Bacterial agent of B5.
Another object of the present application is to provide the above Lactobacillus reuteriLactobacillus reuteri) Application of B5.
Lactobacillus reuteri of the applicationLactobacillus reuteri) B5, the strain is preserved in China general microbiological culture Collection center (CGMCC) for 7 and 18 months of 2022, and has a preservation address of 1 st Xila No. 3, 3 rd national institute of microbiology, academy of China academy of sciences, and is classified and named as Lactobacillus reuteriLactobacillus reuteriThe preservation number is CGMCC No.25324.
The application provides a lactobacillus reuteri-containing composition(Lactobacillus reuteri) The microbial agent of B5 can be liquid microbial agent or solid microbial agent, and is prepared by adding auxiliary materials allowed in the field of microbial preparations by adopting a conventional technical means.
The application provides lactobacillus reuteri(Lactobacillus reuteri) B5 is used for preparing a preparation for the following purposes,
(1) Improving intestinal health;
(2) Improving blood and testis metabolome;
(3) Increasing sperm concentration in male animals;
(4) Increasing sperm motility of male animals;
(5) Improving the mating success rate of male animals and the pregnancy rate of mating female animals; and/or
(6) Improving the number of females producing animals mating with males.
The lactobacillus reuteri LRB5 can be planted in an animal body, and has strong tolerance to artificial bile salts, artificial gastric acid and artificial intestinal juice. The lactobacillus reuteri LRB5 can improve intestinal health, improve intestinal immunity, and increase sperm motility, concentration, pregnancy rate and number of live animals; the strain can improve animal metabolism, increase beneficial metabolites, and increase testis beneficial metabolites such as retinoic acid (regulating spermatogenesis initiation), thereby improving sperm quality and fertility of male animals. The gene sequence of LRB5 and the known lactobacillus reuteri LL7 have a plurality of mutation sites, more than 87% of which are mutated by resequencing. Only 1 strain LRB5 in 15 strains of lactobacillus reuteri isolated and cultured by the method has the effect of improving the semen quality and the fertility, and other strains have no effect.
Drawings
FIG. 1 shows Lactobacillus reuteri of the present applicationLactobacillus reuteri) B5 growth curve (MRS medium);
FIG. 2 shows the body weight change of the test mice in example 2 of the present application at 35 days of the test period;
FIG. 3 shows sperm concentration of test mice in example 2 of the present application;
FIG. 4 shows sperm motility of test mice in example 2 of the present application;
FIG. 5 shows sperm acrosome integrity of the test mice in example 2 of the present application;
FIG. 6 shows the acrosome deformity rate of the test mice in example 2 of the present application;
FIG. 7 shows pregnancy rate of the test mice in example 2 of the present application;
FIG. 8 shows the number of live mice produced in the test mice in example 2 of the present application;
FIG. 9 shows semen collection of a test boar in example 3 of the present application;
FIG. 10 shows the concentration of sperm of a test boar in example 3 of the present application;
FIG. 11 shows the motility of sperm of a test boar in example 3 of the present application;
FIG. 12 shows the acrosome integrity of the sperm of the test boar in example 3 of the present application;
FIG. 13 shows the rate of sperm malformation of the test boar in example 3 of the present application;
FIG. 14 shows the concentration of sperm of the test caprine in example 4 of the present application;
FIG. 15 shows the motility of sperm of a test caprine in example 4 of the present application;
FIG. 16 shows the acrosome integrity of the sperm of the caprine in example 4 of the present application;
FIG. 17 shows the rate of malformation of sperm of the test caprine in example 4 of the present application.
Lactobacillus reuteri of the applicationLactobacillus reuteri) B5 strain, classified and named Lactobacillus reuteriLactobacillus reuteriThe strain is preserved in China general microbiological culture collection center (CGMCC) for 7 months and 18 days in 2022, wherein the preservation address is CGMCC No.25324, and the preservation number is CGMCC No. 3, the microbiological institute of China academy of sciences, which is the North Chen West Lu No. 1, the Korean area of Beijing city.
Detailed Description
The application verifies that the lactobacillus reuteri is%Lactobacillus reuteri) B5 (abbreviated as LRB 5) has beneficial effect on male animal fertility, and a Busulfan (Busulfan) model is used for establishing a mouse oligospermia model, so that the strain can improve sperm concentration, vitality and acrosome integrity, reduce sperm malformation rate, improve success rate of mating a male mouse with a normal female mouse, and pregnancy rate and live number of the female mouse after mating. The semen quality of the boars and the rams is reduced due to the high temperature and high humidity in summer, so that the pregnancy rate and the number of live birth of the sows and the rams are reduced, the sperm concentration and the activity of the boars and the rams in the high temperature and high humidity state in summer can be improved, the acrosome integrity is improved, and the sperm malformation rate is reduced.
EXAMPLE 1 Lactobacillus reuteri ]Lactobacillus reuteri) Isolation of B5
1. Isolation of strains: lactobacillus reuteriLactobacillus reuteriIsolated from the intestinal content. The intestinal contents were taken, 10-fold gradient diluted with sterile normal saline, 0.1mL of the diluted solution of the appropriate dilution was sucked up and evenly spread on BLB agar, and after culturing for 48 hours at 37 ℃, the grown colonies were inoculated into a new BLB solid medium by streaking and purified culture for 48 hours.
2. Enrichment of strains: the purified and cultured strains were inoculated one by one into a sterile MRS broth medium with an inoculating loop on a sterile operating table, placed in a shaking table, cultured at 37℃for 48 hours, subjected to 16S rRNA identification, and the remaining portion was mixed with physiological saline containing 25% glycerol and stored in a-80℃refrigerator.
3. 16S rRNA identification: the enriched bacterial solution is subjected to colony PCR amplification by using bacterial universal primers, and then is sent to a biological engineering (Shanghai) stock company for 16S rDNA sequencing identification, and the 16S rDNA sequences of each strain are compared with the 16S rDNA sequences of all the determined bacteria in a database on NCBI, wherein 6 strains of the bacterial solution and the lactobacillus reuteri are detectedLactobacillus reuteriThe 16S rDNA sequence of HKU 070513 has the highest homology of more than or equal to 99.8 percent, and the category of the 6 strains is determined to be lactobacillus reuteriLactobacillus reuteri。
EXAMPLE 2 Lactobacillus reuteri ]Lactobacillus reuteri)B5 characteristic detection
1. Lactobacillus reuteri @Lactobacillus reuteri) The microbiological properties of B5 are as follows:
(1) Colony morphology: the circular single colony has the diameter of about 1-2 mm, is milky white and opaque, has smooth and slightly convex surfaces and regular edges;
(2) The bacteria form after dyeing presents a short rod shape, has no flagella and can not move;
(3) Growth characteristics: placing in MRS liquid culture medium, shake culturing at 37deg.C, starting logarithmic phase after 5 hr, and entering platform phase for 14 hr, with maximum viable count of 3.1X10 8 CFU/mL. In LBL liquid culture medium, shaking culture at 37deg.C for 4 hr, and loading into logarithmic phase for 12 hr, with maximum viable count of 3.15X10 8 CFU/mL。
2. Bile salt resistance detection
Activated and the concentration of the bacterial liquid is 1 multiplied by 10 8 1mL of the bacterial liquid of CFU/mL is inoculated into 9mL of sterile physiological saline with the concentration of pig bile salt of 0.3g/100mL respectively, and after 1h, the number of viable bacteria is measured by adopting a flat plate coating method.
The results show (Table 1) that LRB5 had the best resistance to bile salts and survived approximately 21.23% at a bile salt concentration of 0.3g/100mL compared to the other 5 strains of Lactobacillus reuteri.
TABLE 1 survival of Lactobacillus reuteri at 0.3% concentration of Strong bile salts%
3. Artificial gastric juice resistance detection
Activated and the concentration of the bacterial liquid is 1 multiplied by 10 8 1mL of each CFU/mL bacterial liquid was inoculated into 9mL of artificial gastric juice with pH=1.5, and after 1 hour, the number of viable bacteria was measured by a plate coating method.
The results showed (table 2) that LRB5 had the best resistance to artificial gastric juice, with a survival rate of about 20.42%, compared to other 5 strains of lactobacillus reuteri after 1h of artificial gastric juice at ph=1.5.
Table 2 survival of lactobacillus reuteri in artificial gastric juice at ph=1.5%
4. Artificial intestinal juice resistant detection
Activated and the concentration of the bacterial liquid is 1 multiplied by 10 8 1mL of each CFU/mL bacterial liquid was inoculated into 9mL of artificial gastric juice with pH=6.8, and after 4 hours, the number of viable bacteria was measured by a plate coating method.
The results showed (Table 3) that LRB5 had the best resistance to artificial intestinal fluid compared with other 5 strains of Lactobacillus reuteri after 4 hours of artificial intestinal fluid, with a survival rate of about 21.56%.
TABLE 3 survival rate of Lactobacillus reuteri in Artificial intestinal juice%
EXAMPLE 3 Lactobacillus reuteri ]Lactobacillus reuteri)B5 effect on fertility in busulfan model Male mice
Determining lactobacillus reuteriLactobacillus reuteri)B5 strain has the best probiotics characteristics, and keeps lactobacillus reuteri @Lactobacillus reuteri)B5 strain was used in subsequent experiments.
1. Experimental method
1. 100 ICR male mice of 21 days old were selected and kept in animal center of animal science institute of national academy of agricultural sciences. In the test process, the temperature of the rat room is controlled at 22+/-2 ℃, the illumination/darkness time is 12 hours, and the rat room can eat and drink water freely.
2. All mice were randomly divided into 5 groups (20 mice per group): control group (Con group), busulfan group (B group), B+LRB5 (10≡6) group, B+LRB5 (10≡8) group, B+LRB5 (10≡10) group. Con group: during the test, the stomach is irrigated once a day with 0.1ml of physiological saline for 14 days; group B: during the test, the stomach is irrigated once a day with 0.1ml of physiological saline for 14 days; B+LRB5 (10≡6): during the test, the concentration of 0.1ml per day of gastric lavage was 1X 10 6 CFU/mL bacterial liquid is 14 days; B+LRB5 (10≡8): during the test, the concentration of 0.1ml per day of gastric lavage was 1X 10 8 CFU/mL bacterial liquid is 14 days; B+LRB5 (10≡10): during the test, the concentration of 0.1ml per day of gastric lavage was 1X 10 10 CFU/mL bacterial liquid, for 14 days. All mice were then kept normally for 21 days (3 weeks) without any treatment, and during the test, the body weight of the mice was measured daily, and the status of the mice, survival conditions, presence or absence of clinical abnormality symptoms, etc. were observed and recorded. After a total of 35 days (14 days+21 days), 10 mice per group were randomly selected on day 36 and samples were collected for analysis. The remaining 10 mice of each group were mated with normal female mice, and each male mouse was mated with 2 female mice for 4 days. Female mice are bred in a single cage after 4 days of mating, and male mice are subjected to sample collection detection and analysis. Pregnant female mice were counted 21 days after mating for number of live mice.
2. Detection index
1. During the test period, the body weight of the mice is counted once every 2 days;
2. measuring the concentration, activity, acrosome integrity and deformity rate of sperm of a male mouse;
3. counting the pregnancy rate and the number of live mice after 21 days of mating;
4. and detecting blood index and testis metabolite of the male mouse.
3. Experimental results
1. Growth conditions: as shown in fig. 2, the change in growth of mice in the whole test resulted in slow weight gain of the mice, and the weight of the busulfan group (B) was lower than that of the control group from the 5 th day of treatment, and was significantly lower than that of the control group until the end of the experiment, whereas the weight of the 3 groups fed LRB5 after busulfan treatment was higher than that of the busulfan group;
2. sperm concentration: as shown in fig. 3, the sperm concentration of the busulfan model group mice was far lower than that of the control group, and the sperm concentration of the 3 groups fed LRB5 after busulfan treatment was far higher than that of the busulfan group, almost reaching the level of the control group.
3. Sperm motility: as shown in fig. 4, the sperm motility of the busulfan model group mice was far lower than that of the control group, and the sperm motility of the 3 groups fed LRB5 after busulfan treatment was far higher than that of the busulfan group, almost reaching the level of the control group;
4. sperm acrosome integrity: as shown in fig. 5, the sperm acrosome integrity of busulfan model group mice was much lower than that of the control group, and the sperm acrosome integrity of the 3 groups fed LRB5 after busulfan treatment was much higher than that of the busulfan group, almost reaching the level of the control group;
5. sperm malformation rate: as shown in fig. 6, the sperm malformation rate of busulfan model group mice was much higher than that of the control group, and the sperm malformation rate of 3 groups fed LRB5 after busulfan treatment was much lower than that of the busulfan group, which was comparable to that of the control group.
6. Pregnancy rate: as shown in fig. 7, the pregnancy rate of the busulfan model group mice after mating with normal female mice was far lower than that of the control group, and the pregnancy rate of the 3 mice fed with LRB5 after busulfan treatment after mating with normal female mice was far higher than that of the busulfan group, almost reaching the level of the control group;
7. number of live births: as shown in fig. 8, after the busulfan model group mice were mated with the normal female mice, the number of live mice produced by the female mice was far less than that of the control group, and after the busulfan treated mice of the 3 groups fed LRB5 were mated with the normal female mice, the number of live mice produced by the female mice was far more than that of the busulfan group, almost reaching the level of the control group.
8. In addition, analysis of the blood and testis metabolites of treated rats found that busulfan increased the harmful metabolites in the blood and testis and decreased beneficial metabolites. And after busulfan treatment, LRB5 is fed to increase blood and testis beneficial metabolites such as glutathione, retinoic acid and total reducing capacity.
EXAMPLE 4 Lactobacillus reuteri ]Lactobacillus reuteri)B5 effect on boar semen quality
1. Experimental method
A40-head (2-3 years old) healthy boar in a boar station is selected, and the weight is about 300 kilograms (kg). All boars were housed in closed pigsty and provided commercial feed and free drinking water, respectively. The pigsty environment is kept clean, ventilated and regularly disinfected. The experiment was 7,8 months in summer.
All boars were randomly divided into 4 groups: control group (Con); LRB5 (10≡9) group; LRB5 (10≡12) group; LRB5 (10≡15); each group had 10 boars. Con group: feeding commercial feed; LRB5 (10≡9) group: commercial feed +10 9 CFU/head/day LRB5; LRB5 (10≡12) group: commercial feed +10 12 CFU/head/day LRB5; LRB5 (10≡15) group: commercial feed +10 15 CFU/head/day LRB5; LRB5 was fed for two weeks (14 days) and then all groups were fed commercial feed for 56 days (total experimental period 70 days), 2 times daily, and dosed. During the test, the status of the boar was observed and recorded. Boar semen was harvested 70 days later and tested for analysis.
2. Detection index
1. During the test period, the signs of the boars are counted every day;
2. semen quality of boars was examined starting on day 71.
3. Experimental results
1. Semen collection amount: as shown in fig. 9, the 3 groups fed LRB5 had more semen collection than the control group;
2. sperm concentration: as shown in fig. 10, the sperm concentration was higher in the 3 groups fed LRB5 than in the control group;
3. sperm motility: as shown in fig. 11, the sperm motility of the 3 groups fed LRB5 was higher than that of the control group;
4. sperm acrosome integrity: as shown in fig. 12, the sperm acrosome integrity of the 3 groups fed LRB5 was higher than the control group;
5. sperm malformation rate: as shown in fig. 13, the sperm malformation rate was lower in the 3 groups fed LRB5 than in the control group.
Example 5 influence of Lactobacillus reuteri LRB5 on semen quality in caprine
1. Experimental method
40 healthy goats (2-3 years old) are selected and used, and the weight of the goat is about 100 kilograms (kg). All goats were housed separately in closed sheep houses and provided with commercial feed (concentrate and fodder) and free drinking water. The sheepfold environment is kept clean, ventilated, and periodically disinfected. The experiment was 7,8 months in summer.
All goats were randomly divided into 4 groups: control group (Con); LRB5 (3 ◊ 10≡8); LRB5 (3 ◊ 10≡11); LRB5 (3 ◊ 10≡14); each group had 10 goats. Con group: feeding commercial feed; LRB5 (3 ◊ 10≡8) group: feeding commercial feed + ◊ 10 8 CFU/head/day LRB5; LRB5 (3 ◊ 10≡11) group: feeding commercial feed + ◊ 10 11 CFU/head/day LRB5; LRB5 (3 ◊ 10≡14) group: feeding commercial feed + ◊ 10 14 CFU/head/day LRB5; LRB5 was fed for two weeks (14 days) and then all groups were fed commercial feed for 56 days (total experimental period 70 days), dosing. During the course of the test, the status of the goats was observed and recorded. Ram semen was collected 70 days later and tested for analysis.
2. Detection index
1. During the test period, signs of the ram were counted daily;
2. semen quality of the ram was measured starting on day 71.
3. Experimental results
1. Sperm concentration: as shown in fig. 14, the sperm concentration was higher in the 3 groups fed LRB5 than in the control group;
2. sperm motility: as shown in fig. 15, the sperm motility of the 3 groups fed LRB5 was higher than that of the control group;
3. sperm acrosome integrity: as shown in fig. 16, the sperm acrosome integrity of the 3 groups fed LRB5 was higher than the control group;
4. sperm malformation rate: as shown in fig. 17, the sperm malformation rate was lower in the 3 groups fed LRB5 than in the control group.
The greatest advantage of LRB5 over lactobacillus reuteri of the same genus is its ability to enhance animal reproductive performance. And has strong tolerance in artificial bile salt, artificial gastric acid and artificial intestinal juice.
The above embodiments are only for explaining the technical solution of the present application, and do not limit the protection scope of the present application.

Claims (4)

1. Lactobacillus reuteri @Lactobacillus reuteri) B5, characterized in that the lactobacillus reuteri @ isLactobacillus reuteri) B5 has the preservation number of CGMCC No.25324.
2. Comprising the Lactobacillus reuteri strain according to claim 1Lactobacillus reuteri) Bacterial agent of B5.
3. The microbial agent of claim 2, wherein the microbial agent is a liquid microbial agent or a solid microbial agent.
4. The lactobacillus reuteri of claim 1Lactobacillus reuteri) B5 is used for preparing the preparation of the following application,
(1) A preparation for improving blood and testis metabolome;
(2) A formulation that increases sperm concentration in a male mammal;
(3) A formulation that increases sperm motility in a male mammal;
(4) A formulation for increasing the success rate of mating in a male animal and the pregnancy rate in a mating female mammal; and/or
(5) A formulation that increases the number of active animals in a female mammal mated with a male.
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