A strain of enteric roseborgia F12-9 and its application

Abstract

The present invention discloses an intestinal Rosenblum bacteria F12-9 and its application. The intestinal Rosenblum bacteria ( Roseburia intestinalis ) F12‑9 strain, whose deposit number is CGMCC NO.30562. The intestinal Roseburia F12‑9 strain provided by the present invention is screened from the rumen contents of adult sheep. It has a strong butyrate production ability, no hemolysis, and strong tolerance. The bacteria can produce a variety of carbohydrate enzymes, which can increase the cellulase activity in the fermentation broth and promote the degradation of feed fiber. As a feed additive, it helps to improve the nutritional value of feed and the digestion and absorption efficiency of animals. At the same time, the bacteria can promote the rumen development and growth ability of young ruminants by increasing fiber digestion and high butyrate production. The present invention is of great significance to the development of animal husbandry.

Description

Technical Field

[0001] The invention relates to the technical field of microbial fermentation, and in particular to a strain of intestinal roseborgia F12-9 with good butyrate production ability and application thereof. Background Art

[0002] As young ruminants grow, their rumen undergoes a physiological function transition from non-ruminant to ruminant. At the same time, the composition of the diet has also changed significantly, gradually transitioning from mainly breast milk to mainly feed on feed, and the digestion method has transitioned from chemical digestion to microbial digestion. As a digestive organ unique to ruminants, the degree of development of the rumen directly affects its production performance in adulthood. Rumen epithelial tissue is an important part of its absorption, transport and metabolic functions. In modern cattle and sheep breeding and production, the rumen development of calves and lambs has great plasticity, and is also easily affected by nutritional regulation and changes in its later production performance. Therefore, regulating the early development of rumen function is the key to ensuring the healthy and efficient production of young animals.

[0003] As a functional microbiome and a candidate for the next generation of probiotics, it plays an important role in promoting gastrointestinal development and enhancing the body's immunity, and has broad application prospects and development potential. However, since most rumen bacteria are strictly anaerobic, the isolation, screening and culture conditions are harsh, and there are few reports of strains that can be used in actual production. Therefore, isolating and screening functional strains from the rumen of ruminants is of great significance for enriching strain resources and promoting the green and healthy development of feed processing and aquaculture. Summary of the invention

[0004] The present invention provides a butyrate-producing intestinal Roseburia F12-9 and an application thereof. The strain has excellent butyrate-producing ability and is tolerant to sodium chloride, bile salts and low pH environments. The bacterial liquid thereof can significantly increase the activities of various carbohydrate enzymes and promote the decomposition and absorption of feed. After feeding young ruminants, the strain can significantly increase their digestion and growth abilities and promote rumen development.

[0005] In a first aspect, the present invention provides a butyrate-producing intestinal Roseborgia ( Roseburia intestinalis ) F12-9, deposited in the General Microbiology Center of China Culture Collection Administration (Address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Postal Code: 100101), with the deposit number CGMCCNO.30562, the deposit date: May 9, 2024, and the classification name: Roseburia intestinalis.

[0006] Roseburia rosea ( Roseburia intestinalis) F12-9 is extracted from the fresh rumen contents of adult sheep. It is tolerant to sodium chloride, bile salts and low pH environment, and the butyrate production reaches 3.29±0.38g / L. Butyrate can supply energy to the body through oxidation in the body. It is the main energy source for the rumen and intestinal epithelial cells of ruminants, and has the functions of regulating digestive health, preventing diarrhea, and enhancing immunity.

[0007] In a second aspect, the present invention provides the above-mentioned intestinal Roseborgia ( Roseburia intestinalis ) The culture method of F12-9 is as follows: activate the intestinal Roseburia ( Roseburia intestinalis ) F12-9 was inoculated into anaerobic fortified Clostridium liquid culture medium with an inoculation amount of 5% to 9%, a growth temperature of 36°C to 38°C, a growth pH of 6 to 8, and a culture time of 8 to 10 hours.

[0008] Preferably, the culture method is to activate the intestinal Roseburia ( Roseburia intestinalis ) F12-9 was inoculated into anaerobic fortified Clostridium liquid culture medium with an inoculation size of 5%, a growth temperature of 37°C, a growth pH of 7, and a culture time of 9 hours.

[0009] In a third aspect, the present invention provides a microbial agent, wherein the microbial agent comprises the above-mentioned intestinal Roseborgia ( Roseburia intestinalis ) F12-9 or its freeze-dried bacterial cells or the bacterial solution obtained by the above-mentioned culture method or the supernatant after centrifugation of the bacterial solution, or the precipitate resuspension after centrifugation of the bacterial solution.

[0010] The microbial agent contains intestinal Roseborgia ( Roseburia intestinalis )F12-9 live bacteria content ≥1.0×10 8 cfu / mL.

[0011] Preferably, the freeze-drying lyophilization protective agent is one or a combination of skimmed milk powder, dextrin, lactose and trehalose.

[0012] In a fourth aspect, the present invention provides the above-mentioned intestinal Roseborgia ( Roseburia intestinalis ) Use of F12-9 or its freeze-dried bacteria, or the bacterial solution obtained by the above-mentioned culture method, or the supernatant after centrifugation of the bacterial solution, or the precipitate resuspension after centrifugation of the bacterial solution, or the above-mentioned microbial agent in the preparation of animal feed additives.

[0013] The present invention also provides a feed additive, which contains the intestinal Roseborgia ( Roseburia intestinalis ) F12-9 or its freeze-dried cells, or the bacterial solution obtained by the above-mentioned culture method, or the supernatant after centrifugation of the bacterial solution, or the precipitate resuspension after centrifugation of the bacterial solution, or the above-mentioned microbial agent.

[0014] In the above feed additives, enteric Roseburia Roseburia intestinalis )F12-9 live bacteria content ≥1.0×10 8 cfu / mL.

[0015] In a fifth aspect, the present invention provides the above-mentioned intestinal Roseborgia ( Roseburia intestinalis ) Use of F12-9 or its freeze-dried bacteria, or the bacterial solution obtained by the above-mentioned culture method, or the supernatant after centrifugation of the bacterial solution, or the precipitate resuspension after centrifugation of the bacterial solution, or the above-mentioned microbial agent, or the above-mentioned feed additive in improving the activity of carbohydrate enzymes and feed degradation rate.

[0016] The present invention further provides a carbohydrate enzyme activity promoter, which contains the above-mentioned intestinal Roseborgia ( Roseburia intestinalis ) F12-9 or its freeze-dried cells, or the bacterial solution obtained by the above-mentioned culture method, or the supernatant after centrifugation of the bacterial solution, or the precipitate resuspension after centrifugation of the bacterial solution, or the above-mentioned microbial agent.

[0017] Among the above carbohydrate enzyme activity promoters, intestinal Roseburia ( Roseburia intestinalis )F12-9 live bacteria content ≥1.0×10 8 cfu / mL.

[0018] The present invention also provides the above-mentioned intestinal Roseborgia ( Roseburia intestinalis ) F12-9 or its freeze-dried bacteria, or the bacterial solution obtained by the above-mentioned culture method, or the supernatant after the bacterial solution is centrifuged, or the precipitate resuspension after the bacterial solution is centrifuged, or the above-mentioned microbial agent, or the above-mentioned feed additive in promoting the growth ability of young ruminants, improving digestion ability, and promoting rumen development.

[0019] The young ruminant is a lamb or a calf.

[0020] The present invention also provides a method for promoting the growth and development of young ruminants, specifically comprising: introducing the intestinal Roseborgia ( Roseburia intestinalis ) F12-9 or its freeze-dried bacterial cells, or the bacterial solution obtained by the above-mentioned culture method, or the supernatant after the bacterial solution is centrifuged, or the precipitate resuspension after the bacterial solution is centrifuged, or the above-mentioned microbial agent, or the above-mentioned feed additive, or the above-mentioned carbohydrate enzyme activity promoter is fed to young ruminants;

[0021] The young ruminant is a lamb or a calf.

[0022] The beneficial effects of the present invention are:

[0023] (1) Intestinal Roseburia in the present invention ( Roseburia intestinalis) F12-9 has excellent butyrate production ability. When cultured at 37℃ in anaerobic enhanced Clostridium liquid culture medium, its butyrate production can reach 3.29±0.38g / L.

[0024] (2) Intestinal Roseburia in the present invention ( Roseburia intestinalis ) F12-9 is tolerant to simulated gastric juice and pancreatic juice environments. Its survival rate is as high as 63.88±3.16% under 6.0% NaCl conditions; 200.82±11.19% under 3.0g / L bile salt conditions; and 100.18±8.44% under pH 3.0 conditions. The above tolerance enables the strain to reproduce better in the animal gastrointestinal environment.

[0025] (3) Intestinal Roseburia in the present invention ( Roseburia intestinalis ) F12-9 bacterial solution can improve the activity of various carbohydrate enzymes, among which carboxymethyl cellulase, xylanase and amylase are particularly effective ( P <0.05), among which the activity of carboxymethyl cellulase increased by 82.9%, the activity of xylanase increased by 74.6%, and the activity of amylase increased by 324%; it can promote the degradation of TMR feed, especially the degradation of dry matter, crude protein, neutral detergent fiber and acid detergent fiber, among which the degradation rate of dry matter increased by 5.5%, the degradation rate of crude protein increased by 6.9%, the degradation rate of neutral detergent fiber increased by 8.2%, and the degradation rate of acid detergent fiber increased by 7.7%. This degradation effect helps to improve the nutritional value of feed and the digestion and absorption efficiency of animals.

[0026] (4) Intestinal Roseburia in the present invention ( Roseburia intestinalis ) F12-9 and its products can significantly increase the growth rate of young ruminants after feeding, with the final weight increased by up to 16.9% after 35 days and the daily weight gain increased by up to 31.2%; increase the TVFA (total volatile fatty acids) and butyric acid content in the rumen and reduce the ammonia nitrogen content, with the rumen ammonia nitrogen content reduced by up to 32.9%, the TVFA content increased by up to 20%, and the butyric acid content increased by up to 258.0%; promote rumen development, with the rumen weight increased by up to 40.8%, the rumen papilla height increased by up to 30.8%, the rumen papilla width increased by up to 46.9%, the rumen papilla density increased by up to 13.8%, and the rumen epithelial thickness increased by up to 39.1%. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0028] Figure 1 This is a phylogenetic tree constructed based on the 16SrRNA gene sequence of the 9 butyrate-producing bacteria in Example 1.

[0029] Figure 2 The colony morphology and Gram staining of intestinal Roseburia F12-9 in Example 1 are shown.

[0030] Figure 3 This is a single factor growth characteristic diagram of intestinal Roseburia F12-9 in Example 2.

[0031] Figure 4 This is a graph showing the optimization results of the inoculation amount of intestinal Roseburia F12-9 in Example 4.

[0032] Figure 5 This is a graph showing the results of optimizing the culture time of enteric Roseburia F12-9 in Example 4.

[0033] Figure 6 This is a graph showing the pH optimization results of enteric Roseburia F12-9 in Example 4. DETAILED DESCRIPTION

[0034] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the drawings of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0035] Example 1 Enteric Roseburia ( Roseburia intestinalis )Screening and identification of F12-9

[0036] In the anaerobic workstation, 2 g of sheep rumen content sample was taken and immersed in 10 mL of sterile anaerobic PBS, vortexed at 3000 r / min for 5 min, and repeated vortexing 3 times. Then 100 μL of rumen content sample was taken for concentration gradient dilution (10 -4 -10 -7), three replicates for each concentration gradient, spread on a fortified Clostridium solid medium, and invert and culture at 37°C. After 48 hours, colonies with different morphological characteristics were picked for streaking purification. After at least 2 purifications, the primary screening strains were obtained.

[0037] A single colony was picked and cultured in anaerobic fortified Clostridium liquid medium for 48 hours. The strain DNA was extracted using lysis buffer, and the 16SrRNA gene of the strain was amplified by PCR using bacterial universal primers 27F (5′-AGAGTTTGATCCTGGCTCAG-3′) and 1492R (5′-GGTTACCTTGTTACGACTT-3′). The PCR product was identified by first-generation sequencing. The effective 16SrRNA gene sequence results were submitted to NCBI for homology comparison analysis in the GenBank database. Generally, the similarity of the comparison results >99% was the same species, and the similarity of 95%-99% was the same genus. According to the identification results, typical butyrate-producing bacteria were screened and a phylogenetic tree was constructed using MEGA-X software ( Figure 1 ).

[0038] A single colony of the screened butyrate-producing strain was selected and inoculated into an anaerobic fortified Clostridium liquid culture medium and cultured statically for 12 h as a seed culture medium ( OD 600 =1.0), the seed liquid was inoculated with 3% of the inoculation amount into the anaerobic fortified Clostridium liquid culture medium, and the butyric acid content was detected by high performance liquid chromatography at 37°C for a certain period of time. An optimal strain was selected based on the acid production content. The target strain was Roseburia ( Roseburia intestinalis ) F12-9, whose butyrate production was significantly higher than that of other butyrate-producing bacteria strains ( P <0.05), and the butyrate production was 3.29±0.38 g / L (Table 1).

[0039]

[0040] Perform Gram staining on the target strain and observe its morphology using an oil immersion microscope ( Figure 2 ). The microbiological characteristics of enteric Roseburia F12-9 are: strict anaerobic bacteria, the colony surface is smooth, the edges form white or milky protrusions, and the shape is round; Gram-positive bacteria, slightly curved rod-shaped.

[0041] Example 2 Enteric Roseburia ( Roseburia intestinalis )F12-9 tolerance test

[0042] A single colony of the intestinal Roseburia F12-9 strain was picked and inoculated into an anaerobic fortified Clostridium liquid culture medium and cultured statically for 12 hours as a seed culture medium (OD600nm=1.0).

[0043] NaCl tolerance test: The seed solution was inoculated with 3% of the inoculum into anaerobic RMC liquid medium with NaCl concentrations of 1.0, 2.0, 3.0, 4.0, 5.0, and 6.0% at 37°C for anaerobic culture. The plate colony counts were performed at 0h and 12h. The test was repeated three times and the survival rate (%) was calculated.

[0044] Bile salt tolerance test: The seed liquid was inoculated with 3% of the inoculum into anaerobic fortified Clostridium liquid medium with bile salt concentrations of 0.3, 0.5, 1.0, 1.5, and 3.0 g / L, and cultured anaerobically at 37°C. The plate colony counts were performed at 0h and 10h, respectively. The test was repeated three times and the survival rate (%) was calculated.

[0045] Gastric and pancreatic juice tolerance test: Artificial gastric juice with pH = 2.5 and 3.0 and artificial pancreatic juice with pH = 8.0 were prepared according to the "Pharmacopoeia of the People's Republic of China (2020 Edition)", and the seed liquid was inoculated into artificial gastric juice and artificial pancreatic juice at a 3% inoculation rate for constant temperature anaerobic culture at 37°C. Plate colony counts were performed at 0h and 5h respectively. The test was repeated three times and the survival rate (%) was calculated.

[0046] Survival rate (%) = final number of viable bacteria measured / initial number of viable bacteria measured × 100.

[0047] The results are as follows Figure 3 The survival rate of enteric Roseburia F12-9 in 6.0% NaCI was as high as 63.88±3.16%. The strain showed good tolerance to 3.0g / L bile salt (200.82±11.19%). The survival rates of the strain in simulated gastric fluid (pH=2.5, pH=3.0) and pancreatic juice were 110.11±9.44%, 98.18±8.44%, and 175.05±5.20%, respectively.

[0048] Example 3 Enteric Roseburia ( Roseburia intestinalis ) Optimization of F12-9 culture conditions

[0049] A single colony of the intestinal Roseburia F12-9 strain was selected and inoculated into an anaerobic fortified Clostridium liquid culture medium and cultured statically for 12 h as a seed culture medium ( OD 600 =1.0).

[0050] Determination of the optimal inoculum size: The seed solution was inoculated into the anaerobic enhanced Clostridium liquid culture medium at 1%, 3%, 5%, 7%, and 9% respectively, and cultured at 37°C. The absorbance of the bacterial solution at 600 nm was measured after 24 hours. OD 600 .

[0051] Determination of the optimal pH: Inoculate the seed liquid at a 3% inoculum into anaerobic fortified Clostridium liquid culture medium with pH values ​​of 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, and 9.0, and culture at 37°C. Measure the absorbance of the bacterial liquid at 600 nm after 10 hours. OD 600 .

[0052] Determination of the optimal time: Inoculate the seed liquid with a 3% inoculation rate into the anaerobic fortified Clostridium liquid culture medium at 37°C for anaerobic culture. Measure the absorbance of the bacterial liquid at 600nm at 2h, 4h, 6h, 8h, 10h, and 12h. OD 600 .

[0053] The results are as follows Figure 4-6 As shown, enteric Roseburia ( Roseburia intestinalis ) F12-9 OD 600 It increases with the increase of inoculum amount. When the inoculum amount is 7%, OD 600 It reaches a maximum of 1.42 and then decreases ( Figure 4 ). With the increase of culture time, the OD 600 It showed an upward trend. At 2h, OD 600 is 0.66, and reaches a maximum value of 1.48 at 10h ( Figure 5 ). When the pH value is 2-5, the intestinal Roseburia F12-9 OD 600 is 0.31 or 0.30. When the pH value is 7.0, OD 600 Reaching a maximum value of 1.69 ( Figure 6 ).

[0054] Example 4 Enteric Roseburia ( Roseburia intestinalis ) F12-9 orthogonal test optimization

[0055] Based on the above single factor test results, a three-factor three-level orthogonal test was conducted on the three single factors of inoculation amount, time, and pH, as shown in Table 2, with three replicates set for each group. The inoculation amount, pH, and time were used as the variation factors, and the absorbance of the bacterial solution ( OD 600) was used as an indicator, and a 3-factor 3-level orthogonal test was used to optimize the culture conditions, with 3 replicates set for each group. The results of the range analysis (Table 3) show that the primary and secondary relationships affecting the growth of intestinal Rosenbjörn F12-9 are pH>time>inoculation amount, indicating that pH is the main factor affecting the growth of intestinal Rosenbjörn F12-9. The optimal culture conditions for intestinal Rosenbjörn F12-9 were determined by the size of the k value as 5% inoculation amount (k=0.767), 9h time (k=0.861), and pH 7 (k=1.107).

[0056]

[0057]

[0058] The results of variance analysis (Table 4) showed that pH and time had a very significant effect ( P <0.01), and the inoculation size had no significant effect. The suitable culture conditions for intestinal Rosenbjöller F12-9 were as follows: inoculate intestinal Rosenbjöller F12-9 activated to the logarithmic phase into anaerobic fortified Clostridium liquid culture medium, with an inoculation size of 5%, pH 7, and a culture time of 9 h.

[0059]

[0060] Example 5 Enteric Roseburia ( Roseburia intestinalis ) TMR feed treatment experiment of F12-9

[0061] Take 18 150mL graduated glass syringes and divide them into 3 groups, 6 in each group. The control group (normal saline) and the experimental group were added with 1.0×10 9 cfu / mL of enteric Roseborgia F12-9 liquid 0.2mL (experiment one) and 2mL (experiment two). Each syringe was placed with 200mg of TMR feed (dry matter mass percentage: 25% silage corn, 15% corn stalks, 35% corn, 10% soybean meal, 10% bran, 5% premix) sample (dried at 60℃ for 48 hours, crushed through a 1mm sieve), 30mL of artificial saliva, and 2mL of distilled water, 1.8mL of distilled water + 0.2mL of target bacterial agent, or 2mL of target bacterial agent. After applying an appropriate amount of vaseline on the front of the syringe piston, insert the syringe to form a closed space. The whole process was completed in a 39℃ water bath with CO2 gas. The above syringes were quickly transferred to an artificial rumen incubator (DSH2-300A, Suzhou Peiying Experimental Equipment Co., Ltd.) for culture. After 24 hours of culture, the syringe was removed and placed in an ice water bath to terminate the fermentation. The fermentation liquid in the culture tube was centrifuged at 1000 g for 10 minutes at 4 °C. Screening and identification of anaerobic fungi isolated from yak rumen with high production of fiber-degrading enzymes[J]. Journal of China Agricultural University, 2010, 15 (3):70-74) was used to determine the carbohydrate degrading enzyme activity of the supernatant (Table 5), and the residue was used to determine the nutrient degradation rate (Table 6) according to the method described in (Effects of three sheep-derived probiotics on rumen in vitro fermentation parameters and nutrient degradation rate [J]. Journal of Animal Nutrition, 2024, 36(06): 4014-4021).

[0062] As shown in Table 5, different concentrations of intestinal Roseburia had significant effects on the activities of carboxymethyl cellulase, xylanase and amylase ( P <0.05). Compared with the control group, the experimental group significantly increased the activities of xylanase and amylase in the fermentation broth ( P <0.05), the second test group significantly increased the activities of carboxymethyl cellulase, xylanase and amylase in the fermentation broth ( P <0.05), among which the activity of carboxymethyl cellulase increased by 82.9%, the activity of xylanase increased by 74.6%, and the activity of amylase increased by 324%.

[0063]

[0064] As shown in Table 6, intestinal Roseburia can promote the degradation of dry matter, crude protein, neutral detergent fiber and acid detergent fiber in feed, among which the degradation rate of dry matter increased by 5.5%, the degradation rate of crude protein increased by 6.9%, the degradation rate of neutral detergent fiber increased by 8.2%, and the degradation rate of acid detergent fiber increased by 7.7%. This shows that intestinal Roseburia as a feed additive may help to improve the nutritional value of feed and the digestion and absorption efficiency of animals.

[0065]

[0066] Example 6 Enteric Roseburia ( Roseburia intestinalis )F12-9 lamb growth promotion experiment

[0067] A total of 72 21-day-old lambs with similar body conditions were randomly divided into four treatment groups, with three replicates in each treatment group and six lambs in each replicate: the control group (CON) was fed with lactation, and the three experimental groups were supplemented with 1.0×10 7 , 1.0×10 8 , 1.0×10 9 CFU / d of enteric Roseburia F12-9 (the three experimental groups were marked as LL, ML, and HL groups, respectively). The experimental period was 35 days. The weight of lambs in each group was measured before and after the experiment, and the results are shown in Table 7:

[0068]

[0069] As shown in Table 7, the growth performance of lambs fed with three dose gradients was good, and their final weight and daily weight gain were significantly higher than those of the CON group ( P＜ 0.05), the maximum increase in final weight was 16.9%, and the maximum increase in daily weight was 31.2%.

[0070] After the experiment, six healthy sheep with similar weight were selected from each group for slaughter. After slaughter, rumen fluid was collected and pH was measured. Then it was filtered through 4 layers of gauze and placed in 50 mL centrifuge tubes and stored in a -20℃ refrigerator. Hu sheep fattening period Effects of adding yeast culture to diet on apparent digestibility of nutrients and rumen fermentation parameters[J]. Journal of Animal Science and Veterinary Medicine, 2019,50(10):2156-216 ) were used to determine the rumen fermentation parameters (Table 8). 2 ), rinse with saline to remove residues, cut 2cm×2cm epithelial tissue samples and place them in 10% formalin solution, refer to ( Butyrate metabolism in rumen epithelium affected by host and diet regime through regulating microbiota in a goat model[J]. Animal Nutrition, 2024,19:41-55 ) were used to determine the morphological parameters of rumen epithelial tissue (Table 9).

[0071] As shown in Table 8, intestinal Roseburia had a significant effect on rumen ammonia nitrogen content, total VFA and butyrate content ( P <0.05). Compared with the control group, the ML and HL groups significantly reduced the rumen ammonia nitrogen content and increased the rumen total VFA and butyrate content ( P <0.05), the LL group significantly increased the total VFA content ( P <0.05), the rumen ammonia nitrogen content decreased by up to 32.9%, the TVFA content increased by up to 20%, and the butyric acid content increased by up to 258.0%.

[0072]

[0073] As shown in Table 9, intestinal Roseburia has a significant effect on the development of lamb rumen. The rumen weight, rumen papilla height, width and density, and rumen epithelial thickness of lambs in the ML and HL groups were significantly higher than those in the control group ( P＜ 0.05), the rumen weight and rumen epithelial thickness of the LL group were higher than those of the control group ( P＜ 0.05), among which rumen weight increased by up to 40.8%, rumen papilla height increased by up to 30.8%, rumen papilla width increased by up to 46.9%, rumen papilla density increased by up to 13.8%, and rumen epithelial thickness increased by up to 39.1%.

[0074]

[0075] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A strain of enteric Roseburia Roseburia intestinalis ) F12-9, characterized in that, The intestinal Roseburia Roseburia intestinalis ) The F12-9 deposit number is CGMCC NO.30562.

2. The intestinal Roseburia of claim 1 ( Roseburia intestinalis ) A method for culturing F12-9, characterized in that: Roseburia intestinalis activated to logarithmic phase ( Roseburia intestinalis ) F12-9 was inoculated into anaerobic fortified Clostridium liquid culture medium with an inoculation amount of 5% to 9%, a growth temperature of 36°C to 38°C, a growth pH of 6 to 8, and a culture time of 8 to 10 hours.

3. The culture method according to claim 2, characterized in that Roseburia intestinalis activated to logarithmic phase ( Roseburia intestinalis ) F12-9 was inoculated into anaerobic fortified Clostridium liquid culture medium with an inoculation size of 5%, a growth temperature of 37°C, a growth pH of 7, and a culture time of 9 hours.

4. A microbial agent, characterized in that: The microbial agent comprises the intestinal Roseburia ( Roseburia intestinalis )F12-9; The microbial agent contains intestinal Roseborgia ( Roseburia intestinalis )F12-9 live bacteria content ≥1.0×10 8 cfu / mL.

5. The intestinal Roseburia of claim 1 ( Roseburia intestinalis ) Use of the microbial agent described in F12-9 or claim 4 in producing butyric acid or preparing animal feed additives.

6. A feed additive, characterized in that: Containing the intestinal Roseburia of claim 1 ( Roseburia intestinalis ) F12-9 or the microbial agent described in claim 4; The feed additive contains enteric Roseburia ( Roseburia intestinalis )F12-9 live bacteria content ≥1.0×10 8 cfu / mL.

7. The intestinal Roseburia of claim 1 ( Roseburia intestinalis ) Use of the microbial agent described in F12-9 or claim 4 or the feed additive described in claim 6 in improving the activity of carbohydrate enzymes and the degradation rate of feed.

8. A carbohydrate enzyme activity promoter, characterized in that Containing the intestinal Roseburia of claim 1 ( Roseburia intestinalis ) F12-9 or the microbial agent described in claim 4; The carbohydrate enzyme activity promoter intestinal Roseborgia ( Roseburia intestinalis )F12-9 live bacteria content ≥1.0×10 8 cfu / mL.

9. The intestinal Roseburia of claim 1 ( Roseburia intestinalis ) Use of the microbial agent described in F12-9 or claim 4 or the feed additive described in claim 6 in promoting the growth ability of young ruminants, improving digestion ability, and promoting rumen development; The young ruminant is a lamb or a calf.

10. A method for promoting the growth and development of young ruminants, characterized in that: The intestinal Roseburia of claim 1 ( Roseburia intestinalis ) F12-9 or the microbial agent described in claim 4, the feed additive described in claim 6, or the carbohydrate enzyme activity promoter described in claim 8 is fed to young ruminants; The young ruminant is a lamb or a calf.

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