A Ningxiang pig-derived bacterial agent combination for preventing and treating piglet diarrhea and its application

Abstract

The present invention discloses a Ningxiang pig-derived bacterial agent combination for preventing and treating piglet diarrhea and an application thereof. Lactobacillus amylovora (preservation number CCTCC NO: M 2024555) and Lactobacillus plantarum (preservation number CCTCC NO: M 2024556) are isolated from Ningxiang pigs. These two bacterial strains have the characteristics of strong acid and bile salt resistance and acid production ability. In addition, they also have strong antioxidant ability, protect intestinal barrier function, and promote intestinal water and ion absorption, etc., which can reduce the incidence of piglet diarrhea and improve growth performance. The combination of the two bacterial strains can exert a better synergistic effect, and their growth-promoting performance, antioxidant ability and diarrhea-alleviating effects are more significant.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and particularly relates to Ningxiang pig-derived amylovoric lactobacillus, plantarum lactobacillus and a composition thereof, which can be used to prevent and treat diarrhea in weaned piglets and improve growth performance. Background Art

[0002] my country's livestock industry has made significant progress in recent years. However, the impact of the African swine fever and COVID-19 outbreaks since 2020, along with the implementation of a complete ban on antibiotics in feed, has placed higher demands on healthy pig farming and environmental safety. However, according to surveys, diarrhea rates in Chinese piglets exceed 20%, a major cause of stunted growth and high mortality. Diarrhea can lead to reduced piglet survival rates, slowed growth and development, reduced feed conversion rates, and increased use of antibiotics and other medications. Diarrhea causes tens of billions of yuan in economic losses annually, severely hindering the healthy development of my country's pig farming industry.

[0003] The intestinal mucosa is the first line of defense for piglets against diarrhea, and the diverse intestinal microbial colonization of this mucosa is a key factor in determining their resistance to diarrhea. Under normal circumstances, the intestinal microbiome can enhance the host's ability to recognize different bacteria and its antioxidant capacity, strengthen the intestinal mucosal barrier, and promote intestinal water and ion absorption, thereby enhancing piglets' resistance to diarrhea. China boasts a rich resource of indigenous pig breeds, which are known for their strong resistance to stress. Current research indicates that the unique microbial composition of these pig breeds is a key contributor to their disease resistance, providing a natural intestinal microbial resource for the study of novel functional microbiota. The Ningxiang Hua pig, a key local pig breed in Hunan Province, is known for its high reproductive rate, excellent meat quality, and strong disease resistance. Precisely due to these exceptional characteristics, the Ningxiang Pig Breeding Farm in Hunan Province was among the first to receive national-level recognition when the National Base for Local Livestock and Poultry Genetic Resources was established in 2008. However, the high-quality microbial resources within the Ningxiang pig have been under-explored and underutilized. In recent years, studies have found that probiotics isolated from local pig breeds such as Tibetan pigs can effectively reduce diarrhea in piglets. This shows that it is possible to specifically explore and screen probiotic strains with strong acid resistance, bile salt resistance, acid production ability and strong antioxidant ability from local pigs, and apply them to feed to improve the antioxidant capacity and intestinal barrier function of piglets, reduce the incidence of diarrhea in piglets, thereby improving the efficiency of pig farming and promoting the high-quality, efficient and green development of our pig farming industry. Summary of the Invention

[0004] The present invention isolates two strains of bacteria from Ningxiang pigs. These two strains have strong acid production ability, strong acid and bile salt resistance, and antioxidant capacity. They can significantly reduce the incidence of diarrhea in weaned piglets, improve antioxidant capacity and growth performance, and the combination of the two strains can work synergistically to achieve better results.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions:

[0006] The present invention provides a strain of Lactobacillus amylovorus YJ001, which is isolated from fresh pig feces in Ningxiang, with a preservation number of CCTCC NO: M 2024555, a preservation date of March 25, 2024, and a preservation unit of the China Center for Type Culture Collection.

[0007] The present invention provides a Lactobacillus plantarum (Lactiplantibacillus plantarum) YJ002, which is isolated from fresh pig feces in Ningxiang, with a preservation number of CCTCC NO: M 2024556, a preservation date of March 25, 2024, and a preservation unit of the China Center for Type Culture Collection.

[0008] The Lactobacillus amylovora YJ001 and Lactobacillus plantarum YJ002 both have certain acid and bile salt resistance capabilities. The two strains can always maintain a survival rate of more than 90% during treatment at pH 4.0 for 120 minutes and 0.05% bile salt for 90 minutes.

[0009] Both the amylovoric Lactobacillus YJ001 and the plantarum Lactobacillus YJ002 have the ability to produce acetic acid and lactic acid. When a GAM medium supplemented with xylooligosaccharides is used as a fermentation medium, the acetic acid production amounts of the amylovoric Lactobacillus YJ001 and the plantarum Lactobacillus YJ002 are 153.17 μg / mL and 146.22 μg / mL, respectively, and the lactic acid production amounts are 98.92 mg / mL and 100.56 mg / mL, respectively.

[0010] The amylovorax Lactobacillus YJ001 and the plantarum Lactobacillus YJ002 both have strong antioxidant capabilities. The supernatants of the amylovorax Lactobacillus YJ001 and the plantarum Lactobacillus YJ002 are collected respectively and then mixed with 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) and 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) solutions respectively. The scavenging rates of the amylovorax Lactobacillus YJ001 on DPPH and ABTS are 22.16% and 31.66% respectively, and the scavenging rates of the plantarum Lactobacillus YJ002 supernatant on DPPH and ABTS are 24.50% and 37.92% respectively.

[0011] Application of Lactobacillus amylovora YJ001 and Lactobacillus plantarum YJ002 in reducing piglet diarrhea rate and improving growth performance: The separate addition of Lactobacillus amylovora YJ001 and Lactobacillus plantarum YJ002 can increase the daily weight gain and feed conversion rate of piglets, and reduce the incidence of piglet diarrhea by 46.5% and 51.1%, respectively; therefore, Lactobacillus amylovora YJ001 and Lactobacillus plantarum YJ002 can be used to prepare feed additive products for preventing and treating diarrhea in weaned piglets, and can also be used to improve the health status and promote growth of weaned piglets.

[0012] A bacterial agent combination, including Lactobacillus amylovora YJ001 and Lactobacillus plantarum YJ002, significantly improved weight gain and feed conversion in weaned piglets compared to individual strains alone, reduced diarrhea incidence by 70.1%, enhanced antioxidant capacity, and reduced inflammatory responses. Therefore, this bacterial agent combination can be used to prepare a feed additive product for preventing and treating diarrhea in weaned piglets, as well as for improving the health and growth of weaned piglets.

[0013] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0014] The amylovorax Lactobacillus YJ001 and plantarum Lactobacillus YJ002 described in the present invention are both isolated from Ningxiang pigs. Their application on pigs will not have a significant impact on the overall intestinal microbial flora, and their application on piglets is highly safe and has no side effects. The two strains are simple and easy to culture, with low production and preparation costs, and have very good application value and prospects in pig feed. The two strains are both highly acid-resistant and bile-resistant, and can reach the posterior end of the pig intestine through gastric acid and duodenal bile acid to exert their effects, regulate the homeostasis of piglet intestinal microorganisms, improve intestinal barrier function, and reduce the incidence of diarrhea; the two strains have strong acetic acid and lactic acid production capabilities and the ability to scavenge oxygen free radicals, which can effectively improve the antioxidant and anti-inflammatory functions of weaned piglets. The combined application of microbial strains allows the strains to work synergistically and the effect is more significant. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 This is a micrograph of Lactobacillus amylovora YJ001.

[0016] Figure 2 This is a micrograph of Lactobacillus plantarum YJ002.

[0017] Figure 3 The acid and bile salt resistance test results of Lactobacillus amylovora YJ001 and Lactobacillus plantarum YJ002. Figure 3 A is the acid tolerance test of Lactobacillus amylovora YJ001, Figure 3 B is the bile salt tolerance test of Lactobacillus amylovora YJ001, Figure 3 C is the acid tolerance test of Lactobacillus plantarum YJ002, Figure 3 D is the bile salt tolerance test of Lactobacillus plantarum YJ002. DETAILED DESCRIPTION

[0018] In order to better understand the present invention, the content of the present invention is further illustrated below in conjunction with the examples, but the content of the present invention is not limited to the following examples.

[0019] Example 1 Isolation and Identification of Lactobacillus amylovora YJ001 and Lactobacillus plantarum YJ002

[0020] In June 2023, Ningxiang pigs were dissected in Ningxiang City, Hunan Province. 0.5 g of the intestinal contents of Ningxiang pigs were immersed in 5 mL of PBS (deoxygenated), vortexed for 5 min, and treated with low-intensity ultrasound for 5 min. The turbid liquid was filtered through a cell sieve to remove solids. The intestinal bacteria of Ningxiang pigs were isolated by gradient dilution. The filtrate was diluted 10 times and 10 -4 to 10 -8 After vortex mixing, 100 μL of the gradient dilution solution was aspirated and spread on a plate, with three replicates for each gradient; the culture was inverted and anaerobic at 37°C (the mixed gas ratio was N2 85%, H2 10%, CO2 5%); the isolated single colonies were purified by streak separation.

[0021] The colony of strain YJ001 is nearly circular, translucent in the early stage and opaque white in the later stage. It has a smooth and dense surface, irregular edges, a convex center, and a diameter of 1-3 mm. Under an optical microscope, YJ001 bacteria appear as typical slender rods, sometimes arranged in clusters, and sometimes as non-branched bacteria with a bulge at one end (such as Figure 1 As shown). The colony of strain numbered YJ002 is round, opaque white, with a smooth and dense surface, smooth edges, a convex center, and a diameter of 1-3 mm. Under an optical microscope, YJ002 bacteria appear as long rods, short rods, and sometimes rod-shaped non-mycobacteria (such as Figure 2 shown).

[0022] The 16S rRNA genes of YJ001 and YJ002 were amplified by PCR and sequenced. The resulting DNA fragments were ligated to the T-vector using a T-vector kit. Positive clones were selected and sent to Beijing Qingke Biotechnology Co., Ltd. for sequencing. The sequences are shown in SEQ ID NO: 1 and SEQ ID NO: 2. Sequence similarity comparisons were performed on EzbioCloud, revealing that strain YJ001 shared 99.8% similarity with Lactobacillus amylovorus, and strain YJ002 shared 99.8% similarity with Lactiplantibacillus plantarum.

[0023] Combined with the morphological characteristics and the results of 16S rRNA gene sequencing and sequence alignment analysis, the isolated strain YJ001 was identified as Lactobacillus amylovorus, and the isolated strain YJ002 was identified as Lactobacillus plantarum. The applicant submitted the samples to the China Center for Type Culture Collection (CCTCC) for preservation on March 25, 2024. The preservation number of Lactobacillus amylovorus YJ001 is CCTCC NO: M 2024555, and the preservation number of Lactobacillus plantarum YJ002 is CCTCC NO: M 2024556. The deposit address is Wuhan University, Wuhan, China.

[0024] Example 2 Acid and bile salt resistance experiments of Lactobacillus amylovora YJ001 and Lactobacillus plantarum YJ002

[0025] 1. Pick freshly activated single colonies of strains YJ001 and YJ002 and inoculate them into 5 mL of liquid culture medium. Incubate at 37°C for 48-72 hours until the stationary phase. The liquid culture medium is the optimal culture medium for the corresponding strain. The optimal culture medium for Lactobacillus amylovora YJ001 is composed of 10.0 g of peptone, 8.0 g of beef extract, 4.0 g of yeast extract, 20.0 g of glucose, 1.0 g of Tween 80, 0.15 g of MgSO4·7H2O, 2.0 g of K2HPO4, 0.06 g of MnSO4·7H2O, 4.0 g of sodium acetate, and 2.0 g of diammonium hydrogen citrate. Adjust the pH to 5.9, add 1 L of distilled water, and sterilize at 121°C for 15 minutes. The optimal culture medium for Lactobacillus plantarum YJ002 is composed of 10.0 g peptone, 8.0 g beef extract, 4.0 g yeast extract, 20.0 g glucose, 1.0 g Tween 80, 0.1 g MgSO4·7H2O, 2.0 g K2HPO4, 0.04 g MnSO4·7H2O, 5.0 g sodium acetate, and 1.8 g diammonium hydrogen citrate. The pH is adjusted to 5.7, 1 L of distilled water is added, and the mixture is sterilized at 121°C for 15 minutes. The mixture is transferred to 5 mL of liquid culture medium at a volume ratio of 1:100 and incubated at 37°C for 12-24 hours with static or shaking. The bacterial solution is then mixed using a vortexer and 800 μL of the solution is pipetted into 1.5 mL sterile centrifuge tubes. The tubes are washed twice with PBS and then resuspended in 800 μL of PBS.

[0026] 2. Acid tolerance: Resuspend the cells in 800 μL PBS and medium with pH 3.0 and 4.0, respectively. Incubate at 37°C and start timing. Perform 10× gradient dilutions on the cells in medium with pH 3.0 and 4.0 at 30 min, 60 min, 90 min, and 120 min, respectively. Take 10 -5 The dilutions were mixed by vortexing, and 100 μL was taken and spread on the plate, and repeated three times; Bile salt tolerance: 800 μL of PBS, 0.05% and 0.1% bile salt concentrations were used to resuspend the bacteria, incubated at 37°C and started timing. At 30 min, 60 min and 90 min, the bacteria treated with 0.05% and 0.1% bile salt concentrations were diluted and spread on the plate, and 10 -5 After mixing the gradient dilutions by vortexing, 100 μL was taken and spread on the plate, and repeated three times; at 0 min, the bacteria resuspended in PBS were diluted 10 times, and 10 -5 After mixing the gradient dilutions on a vortexer, 100 μL was spread on a plate, with three replicates for each gradient. The plates were incubated anaerobically at 37°C for 24 h, and the colonies on the plates were counted.

[0027] The test results are as follows: Figure 3 As shown, Lactobacillus amylovorus YJ001 and Lactobacillus plantarum YJ002 maintained survival rates exceeding 90% after treatment at pH 4.0 for 120 minutes and at 0.05% bile salt for 90 minutes. This indicates that Lactobacillus amylovorus YJ001 and Lactobacillus plantarum YJ002 possess excellent acid and bile salt tolerance, enabling them to survive oral administration through gastric acid and bile acids while maintaining a high survival rate, thus establishing a foundation for intestinal colonization. Therefore, these two strains can be directly added to piglet feed, where they can reach and function in the piglet's intestines, demonstrating their potential for widespread application in practical production.

[0028] Example 3 Experiment on the ability of Lactobacillus amylovora YJ001 and Lactobacillus plantarum YJ002 to scavenge oxygen free radicals

[0029] Single colonies of freshly activated strains YJ001 and YJ002 were picked and inoculated into MRS medium, cultured at 37°C for 48-72 hours until the stationary phase, and then centrifuged to obtain the supernatant of sterile cells. Lactobacillus rhamnosus GG and Escherichia coli DH5α were selected as positive and negative controls for the antioxidant properties of the strains to be tested, respectively. 1 mL of freshly prepared 0.2 mM DPPH solution was mixed with 0.8 mL of the strain supernatant and reacted in the dark for 30 minutes. The absorbance at 517 nm of the supernatant of each strain after reaction with the DPPH solution was measured. The blank control tube contained a mixture of MRS medium and DPPH. The DPPH free radical scavenging rate (%) was calculated as follows: (A blank tube - A test tube / A blank tube) × 100%.

[0030] Mix 7mM ABTS with 2.45mM potassium persulfate and store in the dark at room temperature for 16-18 hours. Then dilute the ABTS solution with alcohol to an absorbance of 0.700±0.020 at 734nm. Next, thoroughly mix 100μL of the strain supernatant with 3.0mL of the diluted ABTS solution and measure the absorbance at 734nm. For the blank control tube, thoroughly mix 100μL of culture medium with 3.0mL of the diluted ABTS solution. + The free radical scavenging rate (%) was calculated as follows: (1-A determination tube / A blank tube)×100%.

[0031] The results are shown in Table 1. Lactobacillus amylovora YJ001 and Lactobacillus plantarum YJ002 had scavenging rates of 22.16% and 24.50% for DPPH free radicals, and 31.66% and 37.92% for ABTS free radicals, respectively. These results indicate that the two strains have strong oxygen free radical scavenging abilities.

[0032] Table 1 Oxygen free radical scavenging rate

[0033]

[0034] Example 4 Determination of Acetic Acid and Lactic Acid Production Capacity of Lactobacillus amylovora YJ001 and Lactobacillus plantarum YJ002

[0035] Freshly activated strains YJ001 and YJ002 were inoculated into 5 mL of GAM liquid medium and incubated at 37°C for 48-72 hours until the stationary phase. The strains were then transferred to 5 mL of GAM liquid medium containing 0.5 g xylooligosaccharides per 100 mL at a volume ratio of 1:100 and incubated at 37°C for 48 hours. The supernatant was centrifuged to obtain the supernatant, which was then placed in an anaerobic tube and transferred to a sealed gas chromatography sample vial. Acetic acid content was determined by GC-MS using the external standard method. Lactic acid determination: Freshly activated strains were inoculated into 5 mL of MRS liquid medium and incubated at 37°C for 72 hours. The supernatant was then centrifuged and the lactic acid content was determined using a colorimetric kit.

[0036] Table 2 Acetic acid and lactic acid content

[0037]

[0038] Example 5 Comparative experiment on the application of combined addition of Lactobacillus amylovora YJ001 and Lactobacillus plantarum YJ002 and single strains in preventing diarrhea in piglets

[0039] Experimental plan: 40 healthy piglets weaned at 21 days of age were randomly divided into 4 groups, with 10 piglets in each group. The experimental period was 14 days. The piglets in the control group were fed a basic diet; the piglets in the experimental group 1 were supplemented with Lactobacillus amylovora YJ001, with a final content of 1.0×10 8 cfu / g (YJ001 group); in the experimental group 2, Lactobacillus plantarum YJ002 was added to the piglets, and the final content was 1.0×10 8 cfu / g (YJ002 group); in the experimental group 3, the piglets were fed a basal diet supplemented with a combination of Lactobacillus amylovora YJ001 and Lactobacillus plantarum YJ002, with the contents of the two bacteria being 1.0×10 6 cfu / g (combined group). The basal diet and nutrient levels are shown in Table 3. The weight of the piglets was recorded on the 0th and 14th days of the experiment, and the feed intake and diarrhea of ​​the piglets were also recorded. At the end of the experiment, the daily weight gain, daily feed intake, feed-to-meat ratio and diarrhea index of the piglets were calculated (the sum of diarrhea scores of each group of piglets / (total number of piglets × total number of days)). The diarrhea scoring rules and standards are as follows: dry feces, 1 point; no diarrhea, normal dry feces, 2 points; mild diarrhea, the surface of the feces is slightly wet, semi-dry feces, 3 points; moderate diarrhea, soft feces, slightly watery, 4 points; severe diarrhea, watery feces, 5 points. Scores of 3 points or above are defined as diarrhea, and the diarrhea incidence rate of each group of piglets during the experiment is calculated ((number of diarrhea piglets × number of diarrhea days) / (total number of piglets in each group × total number of experimental days)). On the 14th day of the experiment, venous blood was collected from 10 piglets in each group to measure blood antioxidant indicators, including malondialdehyde, glutathione peroxidase, total antioxidant capacity, and superoxide dismutase.

[0040] Table 3 Basic diet composition and nutritional levels

[0041] name Ratio (%) Nutritional standards corn 57.5 Crude protein, % 17.64 soybean meal 17.8 Digestible energy, MJ / kg 14.57 fishmeal 6.0 wheat bran 5.3 whey powder 5.63 soybean oil 3.0 Lysine 78% 0.8 DL-methionine 99% 0.47 L-Threonine 98.5% 0.4 L-Tryptophan 98% 0.1 choline 0.1 calcium bicarbonate 1.2 Stone powder 0.5 salt 0.3 Premix* 0.9 total 100

[0042] *Note: Each kg of feed contains anhydrous copper sulfate 200 mg, ferrous sulfate monohydrate 3 g, 1% cobalt 5 g, manganese sulfate monohydrate 100 mg, zinc sulfate monohydrate 2.8 g, 1% iodine 200 mg, 1% selenium 300 mg, and multivitamins 3 g. 1. Effects of Probiotics Added Alone and Combined with Two Strains on Growth Performance of Weaned Piglets

[0043] As shown in Table 4, the body weight and average daily weight gain of piglets in groups YJ001 and YJ002 on the 14th day were significantly higher than those in the control group, and the body weight and average daily weight gain of piglets in the combination group on the 14th day were significantly higher than those in the other three experimental groups; the average daily feed intake of piglets in the combination group was significantly higher than that in the other three experimental groups; the feed-to-meat ratio of piglets in groups YJ001 and YJ002 was significantly lower than that of piglets in the control group, and the feed-to-meat ratio of piglets in the combination group was significantly lower than that in the other three experimental groups.

[0044] Table 4 Growth performance

[0045]

[0046] Note: BW, body weight; ADG, average daily gain; ADFI, average daily feed intake; F / G, feed to meat ratio 2. Effects of adding probiotics alone or in combination on the diarrhea index of weaned piglets

[0047] As shown in Table 5, the diarrhea index and diarrhea rate of piglets in groups YJ001 and YJ002 were significantly lower than those in the control group, and the diarrhea index and diarrhea rate of piglets in the combined group were significantly lower than those in the other three experimental groups.

[0048] Table 5 Diarrhea index and diarrhea rate

[0049]

[0050] 3. Effects of adding probiotics alone or a combination of two strains on blood antioxidant indicators in weaned piglets

[0051] As shown in Table 6, the MDA content in the blood of piglets in the YJ001 and YJ002 groups was significantly lower than that in the control group, while the GSH-Px, CAT and SOD activities were significantly higher than those in the control group; the MDA content in the piglets in the combined group was significantly lower than that in the other three experimental groups, while the GSH-Px, CAT and SOD activities were significantly higher than those in the other three experimental groups.

[0052] Table 6 Blood antioxidant index

[0053]

[0054] Note: MDA, malondialdehyde; GSH-Px, glutathione peroxidase; CAT, total antioxidant capacity; SOD, superoxide dismutase

[0055] Example 6 Comparative Experiment on the Application of Combined Addition of Lactobacillus amylovora YJ001 and Lactobacillus plantarum YJ002 to Alleviate Diarrhea Caused by Pathogenic Escherichia coli in Piglets

[0056] Experimental plan: 30 healthy piglets from Duchangda weaned at 21 days of age were randomly divided into three groups, with 10 piglets in each group. The experimental period was 14 days. The control group piglets were fed a basal diet; the model group piglets were fed a basal diet; the experimental group piglets were fed a basal diet supplemented with a combination of Lactobacillus amylovora YJ001 and Lactobacillus plantarum YJ002, with the two bacteria content of 1.0×10 8 The piglets in the model group and the experimental group were gavaged with 6 mL of E. coli K88 (2×10 10 cfu / mL), and the experiment was continued for 5 days, ending on the 14th day. The basal diet and nutrient levels are shown in Table 3. The weight of the piglets was recorded on the 0th and 14th days of the experiment, and the feed intake and diarrhea of ​​the piglets were also recorded. At the end of the experiment, the daily weight gain, daily feed intake, feed-to-meat ratio and diarrhea index of the piglets were calculated (the sum of diarrhea scores of each group of piglets / (total number of piglets × total number of days)). The diarrhea scoring rules and standards are as follows: dry feces, 1 point; no diarrhea, normal dry feces, 2 points; mild diarrhea, the surface of the feces is slightly wet, semi-dry feces, 3 points; moderate diarrhea, soft feces, slightly watery, 4 points; severe diarrhea, watery feces, 5 points. Scores of 3 points or above are defined as diarrhea, and the diarrhea incidence rate of each group of piglets during the experiment is calculated ((number of piglets with diarrhea × number of days with diarrhea) / (total number of piglets in each group × total number of days of the experiment)).

[0057] 1. Effect of combined addition of two bacterial strains on the growth performance of weaned piglets

[0058] As shown in Table 9, the weight, average daily weight gain, average daily feed intake and feed-to-meat ratio of the piglets in the experimental group with the combined addition of the two strains of bacteria were significantly higher than those in the model group challenged with E. coli at the end of the experiment.

[0059] Table 7 Growth performance

[0060]

[0061] Note: BW, body weight; ADG, average daily gain; ADFI, average daily feed intake; F / G, feed to meat ratio 2. Effect of combined addition of two bacterial strains on diarrhea index in weaned piglets

[0062] As shown in Table 10, the diarrhea index and diarrhea rate of piglets in groups YJ001 and YJ002 were significantly lower than those in the control group, and the diarrhea index and diarrhea rate of piglets in the combined group were significantly lower than those in the other three experimental groups.

[0063] Table 8 Diarrhea index and diarrhea rate

[0064]

Claims

1. A bacterial agent combination, characterized in that: By the preservation number of CCTCC NO: M 2024555 Lactobacillus amylovora ( Lactobacillus amylovorus ) YJ001 and Lactobacillus plantarum with a preservation number of CCTCC NO: M2024556 ( Lactiplantibacillus plantarum )YJ002.

2. Use of the bacterial agent combination according to claim 1 in the preparation of a product for preventing and treating diarrhea in weaned piglets caused by a basal diet.

3. Use of the bacterial agent combination according to claim 1 as a piglet feed additive.

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