Bifidobacterium animalis LLM10 and probiotic complex for promoting initial start of replacement gilts and estrus of primiparous sows and application

Abstract

This invention relates to the field of probiotics, specifically to *Lactobacillus enterica* LLM10 and a probiotic compound that promotes the initial initiation of estrus in gilts and primiparous sows, and its application. The *Lactobacillus enterica* of this invention (… Lactobacillus intestinalis LLM10, with accession number CGMCC No. 26850, is formulated into a compound with cinnamon powder and fox nut powder. This compound enhances the colonization of *Lactobacillus enterica* LLM0 in animals and provides stronger tolerance to artificial bile salts, artificial gastric acid, and artificial intestinal fluid. In particular, it significantly improves hormone synthesis and secretion in gilts and primiparous sows, thereby improving follicle development in gilts, promoting earlier initiation of estrus in gilts, facilitating timely estrus after weaning in primiparous sows, increasing estrus rate, pregnancy rate, and the number of live piglets produced.

Description

Technical Field

[0001] This invention relates to the field of probiotics, specifically to Enterobacter LLM10 and a probiotic compound that promotes the initial initiation of estrus in gilts and primiparous sows, and its application. Background Technology

[0002] Ovarian function in sows is one of the key factors affecting their reproductive capacity. When gilts reach a certain stage of development, follicles develop in the ovaries, producing a large amount of estrogen (E2). This estrogen is then fed back to the hypothalamus, releasing luteinizing hormone (LH), thus initiating estrus and ovulation. Gilts generally begin their estrous cycle at 150-210 days of age. However, some gilts fail to initiate estrus by 210 days of age, resulting in delayed estrus, anestrus, or even silent / silent estrus (normal follicle development, but without estrus symptoms such as increased libido, loss of appetite, vulvar swelling and mucus discharge, or even standing still and allowing boars to mount and mate). Gilts with delayed estrus cannot be used in mass production and are therefore culled, resulting in significant economic losses. Some primiparous sows come into estrus promptly after weaning, but others fail to do so, resulting in delayed estrus, which affects the batch production process and may lead to their culling and economic losses.

[0003] Recent studies have found that gut microbiota is closely related to female reproductive health. However, it is unknown whether combining probiotics with prebiotics such as cinnamon and fox nuts can better improve the quality of eggs and fertility in female animals. Summary of the Invention

[0004] The purpose of this invention is to provide an enterobacterial lactobacillus ( Lactobacillus intestinalis ).

[0005] The purpose of this invention is to provide a probiotic compound that promotes the initial initiation of estrus in gilts and primiparous sows.

[0006] Another object of the present invention is to provide the application of the above-mentioned probiotic compound that promotes the initial initiation of estrus in gilts and primiparous sows.

[0007] According to the present invention, *Lactobacillus enterica* ( Lactobacillus intestinalis LLM10, accession number CGMCC No. 26850, accession date March 20, 2023.

[0008] The probiotic complex for enhancing female animal fertility according to the present invention includes the above-mentioned enterobacteria ( Lactobacillus intestinalis LLM10 and cinnamon powder (CCD), and fox nut powder (EFS).

[0009] This invention provides the use of the above-mentioned probiotic compound that promotes the initial initiation of estrus in gilts and primiparous sows in the preparation of formulations for the following purposes.

[0010] (1) Improves the intestinal health of sows;

[0011] (2) Improves the secretion of blood hormones in sows;

[0012] (3) Promotes follicle development in gilts;

[0013] (4) Promote the early start of the initial stage of replacement sows;

[0014] (5) Promote timely estrus in primiparous sows after weaning;

[0015] (6) Increase estrus rate;

[0016] (7) Increase pregnancy rates; and / or

[0017] (8) Increase the number of live offspring.

[0018] The compound of the present invention can better promote the growth of enterobacteria (Lactobacillus) Lactobacillus intestinalis LLM10 colonizes in animals and has a stronger tolerance to artificial bile salts, artificial gastric acid, and artificial intestinal fluid; it can better improve the intestinal health of sows and better enhance the synthesis and secretion of hormones (GnRH, FSH, LH, E2) in gilts and primiparous sows, thereby improving follicle development in gilts, promoting early initiation of estrus in gilts, promoting timely estrus after weaning in primiparous sows, increasing estrus rate, increasing pregnancy rate, and increasing the number of live piglets. Attached Figure Description

[0019] Figure 1 The growth curves (MRS medium) of Lactobacillus enterica LLM10 and LLM07 screened in Example 1 of the present invention and the prepared compound bacterial agent are shown.

[0020] Figure 2 The changes in blood follicle-stimulating hormone (FSH) levels in gilts after 21 days of feeding with the probiotics and compound agent described in this application are shown. a, b, and c indicate significant differences. The same letter indicates no significant difference, while different letters indicate significant differences. p < 0.05.

[0021] Figure 3 The changes in blood luteinizing hormone (LH) levels in gilts after 21 days of feeding with the probiotics and compound agent described in this application are shown. a, b, and c indicate significant differences. The same letter indicates no significant difference, while different letters indicate significant differences. p < 0.05.

[0022] Figure 4The changes in blood estrogen (E2) levels in gilts after 21 days of feeding with the probiotics and compound agent described in this application are shown. a, b, and c indicate significant differences. The same letter indicates no significant difference, while different letters indicate significant differences. p < 0.05.

[0023] Figure 5 The data shows the changes in the initial start-up age of gilts after 21 days of feeding with the probiotics and compound agent of this application. a and b indicate significant differences. The same letter indicates no significant difference, while different letters indicate significant differences. p<0.05.

[0024] Figure 6 The estrus rate of gilts after 21 days of feeding with the probiotics and compound of this application is shown;

[0025] Figure 7 The pregnancy rate of gilts was shown after 21 days of feeding with the probiotics and compound of this application;

[0026] Figure 8 The data shows the number of live offspring produced by gilts after 21 days of feeding with the probiotics and compound agent described in this application. a and b indicate significant differences. The same letter indicates no significant difference, while different letters indicate significant differences. p < 0.05.

[0027] Figure 9 The changes in blood follicle-stimulating hormone (FSH) levels in primiparous sows after feeding them the probiotics and compound agent of this application for 21 days (21 days of lactation) are shown. a, b, and c indicate significant differences. The same letters do not indicate significant differences, while different letters indicate significant differences. p < 0.05.

[0028] Figure 10 The changes in blood luteinizing hormone (LH) levels in primiparous sows after feeding them the probiotics and compound agent of this application for 21 days (21 days of lactation) are shown. a, b, and c indicate significant differences. The same letters do not indicate significant differences, while different letters indicate significant differences. p < 0.05.

[0029] Figure 11 The changes in blood estrogen (E2) levels in primiparous sows after feeding them the probiotics and compound agent of this application for 21 days (21 days of lactation) are shown. a, b, and c indicate significant differences. The same letter indicates no significant difference, while different letters indicate significant differences. p < 0.05.

[0030] Figure 12 The estrus rate of primiparous sows after feeding them the probiotics and compound agent of this application for 21 days (21 days of lactation);

[0031] Figure 13 The pregnancy rate of primiparous sows after feeding them the probiotics and compound of this application for 21 days (21 days of lactation);

[0032] Figure 14 The data shows the number of live offspring produced by primiparous sows after feeding them the probiotics and compound agent of this application for 21 days (21 days of lactation). a and b indicate significant differences. The same letters do not indicate significant differences, while different letters indicate significant differences. p<0.05.

[0033] The enterobacterium of the present invention ( Lactobacillus intestinalis LLM10, classified as Enterobacter. Lactobacillus intestinalis The strain was deposited on March 20, 2023, at the China General Microbiological Culture Collection Center (CGMCC, address: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, 100101, China), with accession number CGMCC No. 26850. Detailed Implementation

[0034] This application isolated 21 strains of *Lactobacillus enterica*, named LLM01, LLM02, LLM03, ..., LLM21. Studies revealed that only LLM10 significantly promoted the initial initiation of estrus in gilts and primiparous sows.

[0035] This invention further verifies the beneficial effects of probiotic compound on promoting early initiation of estrus in gilts and primiparous sows. Using 110-day-old two-way crossbred gilts and primiparous sows (starting from the first day of their first farrowing) as models, it was found that the probiotic compound can significantly increase hormone synthesis and secretion (GnRH, FSH, LH, E2) in gilts and primiparous sows compared to the single strain LLM10. This results in improved follicle development in gilts, promoted early initiation of estrus in gilts, facilitated timely estrus after weaning in primiparous sows, increased estrus rate, increased pregnancy rate, and increased number of live offspring.

[0036] In the following examples, the preparation process of cinnamon powder (Cinnamomum cassia (L.) D. Don; abbreviated CCD) is as follows:

[0037] (1) Collecting cinnamon: Cinnamon bark is collected in February and dried in a cool, shady place.

[0038] (2) Grind into powder slowly and evenly using a grinder;

[0039] (3) Store the obtained cinnamon powder in a cool, dry place for later use.

[0040] The preparation process of Euryale ferox Salisb. (abbreviated EFS) is as follows:

[0041] (1) Collecting Euryale ferox fruits: Harvest Euryale ferox fruits in autumn, wash them clean, and air dry them in a cool place.

[0042] (2) Grind into powder slowly and evenly using a grinder;

[0043] (3) Store the obtained gorgon fruit powder in a cool, dry place for later use.

[0044] The compound microbial agent used in the following examples includes: (strain LLM10 or LLM07) + CCD + EFS. 100 grams of the compound microbial agent includes 20-40 grams (dried strain LLM10 or LLM07), 10-15 grams of cinnamon powder CCD, and 20-45 grams of foxnut powder EFS. 1 gram of the compound agent is dissolved in 100 ml of physiological saline. In the same experiment, the mass of dried strain LLM10 or LLM07, CCD, and EFS used in each parallel treatment (e.g., CCD+EFS group, LLM10 group, LLM10+CCD+EFS group, LLM07 group, LLM07+CCD+EFS group) is the same.

[0045] For example, in the compound microbial agent treatment group (LLM10+CCD+EFS group), 1 gram of compound microbial agent was fed daily, while in the parallel strains LLM10 or LLM07 group and CCD+EFS group, the amount of active ingredient fed was the amount of dry microbial agent or CCD+EFS contained in 1 gram of compound microbial agent. That is, the amount of the same active ingredient was the same in the parallel treatment groups.

[0046] The calculation method for the synergistic effect coefficient:

[0047] Groups: (1) Control group (Con group); (2) CCD+EFS group (CCD+EFS group); (3) LLM10 group (LLM10 group); (4) LLM10+CCD+EFS probiotic compound group (LLM10+CCD+EFS group).

[0048] Calculation formula:

[0049]

[0050] If the synergy coefficient is greater than 100%, it indicates that there is a synergy effect.

[0051] Example 1: Isolation and culture of Lactobacillus LLM10 in probiotic compound

[0052] (1) Strains isolated: Enterobacter ( Lactobacillus intestinalisLLM10 was isolated from intestinal contents. Intestinal contents were serially diluted 10-fold with sterile physiological saline. 0.1 mL of each diluted solution was evenly spread onto MRS agar and incubated at 37°C for 48 h. The resulting colonies were then streaked onto fresh MRS solid medium for purification and incubation for another 48 h. Twenty-one strains of *Lactobacillus enterica* were isolated and named LLM01, LLM02, LLM03, ..., LLM21. Studies showed that only LLM10 significantly promoted the initial initiation of estrus in gilts and estrus in primiparous sows. This application also provides research data on another *Lactobacillus enterica* strain, LLM07.

[0053] (2) Enrichment of strains: The purified strains were inoculated one by one into sterile MRS broth medium on a sterile operating table using an inoculation loop, placed in a shaker, and cultured at 37°C for 48 hours. 16S rDNA identification was performed, and the remaining part was mixed with physiological saline containing 25% glycerol and stored in a -80°C freezer.

[0054] (3) 16S rDNA identification: After enriching the bacterial culture, colony PCR amplification was performed using universal bacterial primers, followed by 16S rDNA sequencing identification. The 16S rDNA sequences of each strain were compared with the 16S rDNA sequences of all identified bacteria in the NCBI database. It was found that strains LLM01–LLM21 were similar to Lactobacillus enterica ( Lactobacillus intestinalis) The 16S rDNA sequences showed the highest homology similarity, with a similarity greater than or equal to 99.9%, thus identifying these strains as Enterobacteriaceae (Enterobacter). Lactobacillus intestinalis) The enterobacterium ( Lactobacillus intestinalis The 16S rDNA of LLM10 is shown below (SEQ ID NO: 1):

[0055]

[0056] Example 2: Characteristic detection of Lactobacillus intestinalis LLM10 and Lactobacillus LLM07 in probiotic compound.

[0057] 1. The microbiological characteristics of *Lactobacillus enterica* LLM10 and LLM07 (two morphologically similar strains) are as follows:

[0058] (1) Colony morphology: round single colonies, about 1.0~1.7 mm in diameter, milky white, opaque, with a smooth surface and slightly raised edges;

[0059] (2) After staining, it appears as a short rod-shaped structure without flagella and cannot move;

[0060] (3) Growth characteristics: such as Figure 1 As shown, LLM10 was inoculated alone in MRS liquid medium and cultured statically at 37°C. It entered the logarithmic growth phase after 4 hours and reached the plateau phase after 12 hours, with a maximum viable count of 2.87 × 10⁻⁶. 9 CFU / mL. The probiotic complex (LLM10+CCD+EFS) was inoculated into MRS liquid medium and incubated statically at 37°C. It entered the logarithmic growth phase after 3 hours and reached the plateau phase after 14 hours, with a maximum viable count of 3.3 × 10⁻⁶ CFU / mL. 9 CFU / mL.

[0061] LLM07 cells were inoculated alone in MRS liquid medium and incubated statically at 37°C. They entered the logarithmic growth phase after 4 hours and reached the plateau phase after 13 hours, with a maximum viable count of 2.65 × 10⁻⁶ cells / year. 9 CFU / mL. LLM07+CCD+EFS was inoculated in MRS liquid medium and incubated statically at 37°C. It entered the logarithmic growth phase after 3 hours and reached the plateau phase after 15 hours, with a maximum viable count of 3.12 × 10⁻⁶. 9 CFU / mL.

[0062] 2. Bile salt tolerance test

[0063] The activated bacterial culture was prepared to a concentration of 1×10⁻⁶. 8 CFU / mL, take 1mL of bacterial suspension containing only LLM10 or LLM07, or 1mL of probiotic compound (LLM10+CCD+EFS), or LLM07+CCD+EFS, and inoculate it into 9mL of sterile physiological saline containing 0.3g / 100mL of porcine bile salts. After 1 hour, determine the viable count using the plate spread method.

[0064] The results (Table 1) showed that *Lactobacillus enterica* LLM10 could grow at a bile salt concentration of 0.3 g / 100 mL, with a survival rate of approximately 26.12%. In the probiotic complex, LLM10 grew better, exhibiting better bile salt tolerance, with a survival rate of approximately 32.24%. LLM07 was similar to LLM10.

[0065] Table 1. Survival rate (%) of different samples at a strong bile salt concentration of 0.3%.

[0066] .

[0067] a and b represent significant differences; the same letters do not indicate significant differences, while different letters indicate significant differences (p < 0.05).

[0068] 3. Test for resistance to artificial gastric fluid

[0069] The activated bacterial culture concentration is 1×10⁻⁶ 8 For each CFU / mL bacterial culture, 1 mL of bacterial culture containing only LLM10 or LLM07, or 1 mL of probiotic complex (LLM10+CCD+EFS), or LLM07+CCD+EFS, was inoculated into 9 mL of artificial gastric fluid at pH 1.5. After 1 hour, the viable count was determined using the plate spread method.

[0070] The results (Table 2) showed that after 1 hour in artificial gastric fluid at pH 1.5, *Lactobacillus enterica* LLM10 exhibited good resistance to artificial gastric fluid, with a survival rate of approximately 20.34%. The probiotic complex-formulated LLM10 showed even better growth and resistance to artificial gastric fluid, with a survival rate of approximately 33.43%. LLM07 was similar to LLM10.

[0071] Table 2. Survival rate (%) of different samples in artificial gastric fluid at pH 1.5

[0072] .

[0073] a and b represent significant differences; the same letters do not indicate significant differences, while different letters indicate significant differences (p < 0.05).

[0074] 4. Resistance to artificial intestinal fluid test

[0075] The activated bacterial culture concentration is 1×10⁻⁶ 8 For bacterial suspensions with CFU / mL, 1 mL of bacterial suspension containing only LLM10 or LLM07, or 1 mL of bacterial suspension containing pro-ovarian compound (LLM10+CCD+EFS), or LLM07+CCD+EFS, was inoculated into 9 mL of artificial gastric fluid at pH 6.8. After 4 hours, the viable bacterial count was determined by plate spread method.

[0076] The results (Table 3) showed that after 4 hours in artificial intestinal fluid, *Lactobacillus enterica* LLM10 exhibited good resistance to artificial intestinal fluid, with a survival rate of approximately 25.27%. The probiotic complex LLM10 showed better growth and resistance to artificial gastric fluid, with a survival rate of approximately 34.96%. LLM07 was similar to LLM10.

[0077] Table 3 Survival rate (%) of different samples in artificial intestinal fluid

[0078] .

[0079] a and b indicate statistical significance. Differences between identical letters are not significant, while differences between different letters are significant (p < 0.05).

[0080] Example 3: Effects of probiotic compound on estrus and reproductive performance of gilts

[0081] I. Experimental Methods

[0082] Three hundred healthy replacement gilts (110 days old) weighing approximately 50 kg were selected. All gilts were housed in enclosed pens, provided with commercial feed and free access to water. The pens were kept clean, well-ventilated, and disinfected regularly.

[0083] All sows were randomly divided into 6 groups: control group (Con), CCD+EFS group, LLM10 group, probiotic complex group (LLM10+ CCD + EFS) group, LLM07 group, and LLM07+ CCD + EFS group; each group consisted of 50 sows. Con group: fed commercial feed; CCD+EFS group: fed commercial feed + CCD+EFS per sow daily; LLM10 group: fed commercial feed + LLM10; probiotic complex group (LLM10+ CCD + EFS): fed commercial feed + LLM10+ CCD + EFS (1 gram of complex per sow daily); LLM07 group: fed commercial feed + LLM07; LLM07+ CCD + EFS group: fed commercial feed + LLM07+ CCD + EFS (1 gram of complex per sow daily). The gilts were fed LLM07 (or LRY09) and a compound supplement for 3 weeks (21 days), after which all groups were fed commercial feed twice daily in fixed quantities. The condition of the gilts was observed and recorded throughout the experiment. Artificial insemination was performed at 230 days of age, with a gestation period of 114 days. The number of live piglets born was then recorded.

[0084] II. Detection Indicators

[0085] 1. During the trial period, the vital signs of the replacement gilts were recorded daily;

[0086] 2. Test the blood hormone levels of gilts: GnRH, FSH, LH, and E2.

[0087] 3. Determine the initial age of replacement gilts;

[0088] 4. Detect the estrus rate of replacement gilts;

[0089] 5. Test the pregnancy rate of gilts;

[0090] 6. Detect the number of live offspring.

[0091] III. Experimental Results

[0092] 1. Blood FSH concentration in gilts: such as Figure 2 As shown, the blood FSH levels of sows fed LLM10 were higher than those of the control group (Con), while the blood FSH levels of the probiotic complex group (LLM10+CCD+EFS) were significantly higher than those of the LLM10 group; LLM10 and CCD+EFS had a synergistic effect, with a synergistic effect coefficient of 234%. However, the two groups fed LLM07 or LLM07+CCD+EFS were similar to the control group (Con) and showed no synergistic effect.

[0093] 2. LH concentration in the blood of gilts: such as Figure 3 As shown, the blood LH levels in the group fed LLM10 were higher than those in the control group (Con), while the blood LH levels in the probiotic compound group (LLM10+CCD+EFS) were significantly higher than those in the LLM10 group; LLM10 and CCD+EFS had a synergistic effect, with a synergistic effect coefficient of 189%. However, the two groups fed LLM07 or LLM07+CCD+EFS were similar to the control group (Con) and showed no synergistic effect.

[0094] 3. Blood E2 concentration in gilts: such as Figure 4 As shown, the blood E2 level of sows fed LLM10 was higher than that of the control group (Con), while the blood E2 level of the probiotic complex group (LLM10+CCD+EFS) was significantly higher than that of the LLM10 group; LLM10 and CCD+EFS had a synergistic effect, with a synergistic effect coefficient of 171%. However, the two groups fed LLM07 or LLM07+CCD+EFS were similar to the control group (Con) and did not have a synergistic effect.

[0095] 4. Initial age of replacement gilts: such as Figure 5As shown, the initial age of sows fed LLM10 was earlier than that of the control group (Con), while the initial age of sows fed the probiotic compound group (LLM10+CCD+EFS) was significantly earlier than that of the LLM10 group; LLM10 and CCD+EFS had a synergistic effect, with a synergistic effect coefficient of 262%. However, the two groups fed LLM07 or LLM07+CCD+EFS were similar to the control group (Con) and did not have a synergistic effect.

[0096] 5. Estrus rate of gilts: such as Figure 6 As shown, the estrus rate in the group fed LLM10 was higher than that in the control group (Con), while the estrus rate in the probiotic compound group (LLM10+CCD+EFS) was significantly higher than that in the LLM10 group; LLM10 and CCD+EFS had a synergistic effect, with a synergistic effect coefficient of 143%. However, the two groups fed LLM07 or LLM07+CCD+EFS were similar to the control group (Con) and did not have a synergistic effect.

[0097] 6. Pregnancy rate of gilts: such as Figure 7 As shown, the pregnancy rate of sows fed LLM10 was higher than that of the control group (Con), while the pregnancy rate of the probiotic compound group (LLM10+CCD+EFS) was significantly higher than that of the LLM10 group; LLM10 and CCD+EFS had a synergistic effect, with a synergistic effect coefficient of 167%. However, the two groups fed LLM07 or LLM07+CCD+EFS were similar to the control group (Con) and did not have a synergistic effect.

[0098] 7. Number of live piglets produced by gilts: e.g. Figure 8 As shown, the number of live offspring produced in the group fed LLM10 was higher than that in the control group (Con), while the number of live offspring produced in the probiotic compound group (LLM10+CCD+EFS) was significantly higher than that in the LLM10 group; LLM10 and CCD+EFS had a synergistic effect, with a synergistic effect coefficient of 140%. However, the two groups fed LLM07 or LLM07+CCD+EFS were similar to the control group (Con) and did not have a synergistic effect.

[0099] Example 4: Effects of probiotic compound on estrus and reproductive performance in primiparous sows

[0100] I. Experimental Methods

[0101] Three hundred healthy primiparous sows, weighing approximately 165 kg, were selected. All sows were housed in enclosed pens, provided with commercial feed and free access to water. The pens were kept clean, well-ventilated, and disinfected regularly.

[0102] All primiparous sows were randomly divided into 6 groups: control group (Con), CCD+EFS group, LLM10 group, probiotic complex group (LLM10+ CCD + EFS) group, LLM07 group, and LLM07+ CCD + EFS group; each group consisted of 50 primiparous sows. Con group: fed commercial feed; CCD+EFS group: fed commercial feed + CCD+EFS per primiparous sow daily; LLM10 group: fed commercial feed + LLM10; probiotic complex group (LLM10+ CCD + EFS): fed commercial feed + LLM10+ CCD + EFS (1 gram of complex per primiparous sow daily); LLM07 group: fed commercial feed + LLM07; LLM07+ CCD + EFS group: fed commercial feed + LLM07+ CCD + EFS (1 gram of complex per primiparous sow daily). The sows were fed LLM07 (or LRY09) and a compound supplement for 3 weeks (21 days; lactation period), after which all groups were fed commercial feed twice daily in fixed quantities. During the experiment, the condition of primiparous sows was observed and recorded. Weaning was performed 21 days after lactation, and the estrus rate was recorded after weaning. Artificial insemination was performed 7 days after weaning, followed by a gestation period of 114 days. The number of live piglets born was then recorded.

[0103] II. Detection Indicators

[0104] 1. During the trial, the physical characteristics of primiparous sows were recorded daily;

[0105] 2. Detect blood hormones GnRH, FSH, LH, and E2 in primiparous sows;

[0106] 3. Determine the initial age of primiparous sows;

[0107] 4. Detect the estrus rate of primiparous sows;

[0108] 5. Detect the pregnancy rate of primiparous sows;

[0109] 6. Detect the number of live offspring.

[0110] III. Experimental Results

[0111] 1. Blood FSH concentration in primiparous sows: such as Figure 9 As shown, the blood FSH levels of primiparous sows fed LLM10 were higher than those of the control group (Con), while the blood FSH levels of the probiotic complex group (LLM10+CCD+EFS) were significantly higher than those of the LLM10 group; LLM10 and CCD+EFS had a synergistic effect, with a synergistic effect coefficient of 307%. However, the two groups fed LLM07 or LLM07+CCD+EFS were similar to the control group (Con) and showed no synergistic effect.

[0112] 2. Blood LH concentration in primiparous sows: such as Figure 10 As shown, the blood LH levels of primiparous sows fed LLM10 were higher than those of the control group (Con), while the blood LH levels of the probiotic complex group (LLM10+CCD+EFS) were significantly higher than those of the LLM10 group; LLM10 and CCD+EFS had a synergistic effect, with a synergistic effect coefficient of 261%. However, the two groups fed LLM07 or LLM07+CCD+EFS were similar to the control group (Con) and showed no synergistic effect.

[0113] 3. Blood E2 concentration in primiparous sows: such as Figure 11 As shown, the blood E2 of primiparous sows fed LLM10 was higher than that of the control group (Con), while the blood E2 of the probiotic complex group (LLM10+CCD+EFS) was significantly higher than that of the LLM10 group; LLM10 and CCD+EFS had a synergistic effect, with a synergistic effect coefficient of 231%. However, the two groups fed LLM07 or LLM07+CCD+EFS were similar to the control group (Con) and did not have a synergistic effect.

[0114] 4. Estrus rate in primiparous sows: such as Figure 12 As shown, the estrus rate of primiparous sows fed LLM10 was higher than that of the control group (Con), while the estrus rate of the probiotic compound group (LLM10+CCD+EFS) was significantly higher than that of the LLM10 group; LLM10 and CCD+EFS had a synergistic effect, with a synergistic effect coefficient of 150%. However, the two groups fed LLM07 or LLM07+CCD+EFS were similar to the control group (Con) and did not have a synergistic effect.

[0115] 5. Pregnancy rate of primiparous sows: such as Figure 13 As shown, the pregnancy rate of primiparous sows fed LLM10 was higher than that of the control group (Con), while the pregnancy rate of the probiotic compound group (LLM10+CCD+EFS) was significantly higher than that of the LLM10 group; LLM10 and CCD+EFS had a synergistic effect, with a synergistic effect coefficient of 138%. However, the two groups fed LLM07 or LLM07+CCD+EFS were similar to the control group (Con) and did not have a synergistic effect.

[0116] 6. Number of live piglets born to primiparous sows: e.g. Figure 14 As shown, the number of live offspring produced in the group of primiparous sows fed LLM10 was higher than that in the control group (Con), while the number of live offspring produced in the probiotic compound group (LLM10+CCD+EFS) was significantly higher than that in the LLM10 group; LLM10 and CCD+EFS had a synergistic effect, with a synergistic effect coefficient of 233%. However, the two groups fed LLM07 or LLM07+CCD+EFS were similar to the control group (Con) and did not have a synergistic effect.

[0117] The above embodiments are only used to understand the technical solutions of this application and do not limit the scope of protection of this application.

Claims

1. Lactobacillus intestinalis (LIM10) characterized in that, Lactobacillus intestinalis The enterobacterium ( Lactobacillus intestinalis The accession number of LLM10 is CGMCC No. 26850. ​ 2. A probiotic complex for promoting initial start of replacement gilts and estrus of primiparous sows, characterized in that, The probiotic complex comprises Lactobacillus intestinalis (LLM10) of claim 1 Lactobacillus intestinalis ) LLM10, cinnamon powder and gordon euryale seed powder.

3. The probiotic compound agent for promoting the initial initiation of estrus in gilts and primiparous sows according to claim 2, characterized in that, Based on 100 grams of the probiotic complex agent, including the Enterococcus faecium (Lactococcus lactis) Lactobacillus intestinalis ) LLM10 dry bacteria 20-40 grams, cinnamon powder CCD 20-30 grams, gordon euryale seed powder EFS 5-30 grams, and the rest is auxiliary materials.

4. The use of the probiotic compound agent according to claim 2 for promoting the initial initiation of estrus in gilts and primiparous sows in the preparation of formulations for the following purposes: (1) Improves the intestinal health of sows; (2) Improves the secretion of blood hormones in sows; (3) Promotes follicle development in gilts; (4) Promote the early start of the initial stage of replacement sows; (5) Promote timely estrus in primiparous sows after weaning; (6) Improve the estrus rate of primiparous sows after weaning; (7) Improve the pregnancy rate of primiparous sows after weaning; and (8) Increase the number of live piglets produced by primiparous sows after weaning.

5. Lactobacillus intestinalis for use according to claim 1, Lactobacillus intestinalis ) Use of LLM10 for the preparation of a formulation for the following uses, (1) Improves the intestinal health of sows; (2) Improves the secretion of blood hormones in sows; (3) Promotes follicle development in gilts; (4) Promote the early start of the initial stage of replacement sows; (5) Promote timely estrus in primiparous sows after weaning; (6) Improve the estrus rate of primiparous sows after weaning; (7) Improve the pregnancy rate of primiparous sows after weaning; and (8) Increase the number of live piglets produced by primiparous sows after weaning.

Images