A method and system for screening of ecological coexistence probiotic

Abstract

The application discloses a screening method and system of ecological coexistence probiotics, and the screening method comprises the following steps: obtaining single strains with high throughput of preset functions based on multi-target high-throughput screening; combining the strains in a functional combination mode to obtain probiotic combinations; in vitro co-culturing, detecting the OD value of the probiotic combinations, and if the OD value of the probiotic combinations is greater than a preset threshold, it is judged that the probiotic combinations have a high possibility of in-vivo synergistic symbiosis, otherwise, the probiotic combinations have a low possibility of in-vivo synergistic symbiosis, and the preset threshold is 1.9-2.1. The screening method combines the probiotic combinations in a functional combination mode, and adopts a threshold method for screening, and the screening is performed according to the principle that the probiotic combinations have a high possibility of in-vivo synergistic symbiosis if the OD value of the probiotic combinations is greater than the preset threshold, so that the screening method and system can quickly screen effective probiotic combinations compared with random combination.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, and more specifically, relates to a method and system for screening eco-coexisting probiotics. Background Technology

[0002] When ingested in sufficient quantities, probiotics can have beneficial health effects on the host. However, single strains often have limited effectiveness. Studies have shown that combining probiotics generally yields better results than single strains, but the effectiveness of probiotic combinations is uncertain. Existing probiotic combinations typically combine 2-3 strains, and the screening methods are often limited. The resulting strains are usually only validated for their individual effects, or are simply random combinations of different strains, offering little reference value for other strain combinations.

[0003] Furthermore, random combinations of bacterial strains do not always produce more beneficial effects. For example, in studies on probiotics improving obesity, while individual administration of VKL / VKB probiotics reduced the body weight of fed mice, their combination did not have a significant anti-obesity effect; even the IMV B-7280 strain alone was found to have better effects than the VKL / VKB / IMV B-7280 combination. This is mainly because different strains may have various interactions, such as metabolic dependence, nutrient competition, and bacteriocin antagonism, which prevents strains from coexisting well in vivo, and may even negate or reduce the beneficial functions of probiotics.

[0004] While functional and in vivo experimental validation of probiotic combinations can alleviate the functional uncertainty caused by arbitrary combinations of probiotics to some extent, this validation process is lengthy and time-consuming, especially for probiotic combinations with novel therapeutic effects, where the workload is enormous and the time required is extensive. Therefore, how to quickly and effectively screen for probiotic combinations that can coexist ecologically and produce significant effects is a pressing problem that needs to be solved. Summary of the Invention

[0005] To address the above-mentioned deficiencies or improvement needs of existing technologies, this invention provides a method and system for screening probiotics that coexist in vivo. The purpose is to combine strains according to functional combinations, and to screen probiotic combinations with an OD value greater than a preset threshold (1.9-2.1) to identify those with a high probability of symbiosis in vivo. This solves the problem of complex and time-consuming operations in existing methods for screening probiotic combinations with good in vivo symbiosis.

[0006] To achieve the above objectives, according to one aspect of the present invention, a method for screening eco-coexisting probiotics is provided, comprising the following steps:

[0007] High-throughput screening based on multiple targets yields single strains with preset functions and high throughput.

[0008] Different high-throughput strains are combined according to their functions to obtain probiotic combinations;

[0009] In vitro co-culture was performed, the OD value of the probiotic combination was measured, and the selection was based on the following principles:

[0010] If the OD value of the probiotic combination is greater than a preset threshold, it is determined that the probiotic combination is likely to coexist synergistically in vivo; otherwise, the possibility of coexisting synergistically in vivo is small. The preset threshold is between 1.9 and 2.1.

[0011] Preferably, in the screening method for the coexisting probiotics, the OD value is specifically determined according to the following method:

[0012] The same probiotic combination was inoculated at a preset ratio and cultured until the logarithmic growth phase. The OD value was then measured under the corresponding wavelength conditions. The preset ratio was 1%-3%.

[0013] Preferably, the method for screening probiotics for ecological coexistence involves selecting probiotic combinations that have the function of weight loss by combining probiotics that have two or more functions, such as pancreatic lipase inhibition, bile salt hydrolase activity, stimulation of GLP-1 hormone secretion, and inhibition of preadipocyte differentiation. The probiotics are then co-cultured in vitro, and the OD value is detected.

[0014] Preferably, the method for screening probiotics for ecological coexistence involves selecting probiotic combinations that have four functions when screening for probiotic combinations that have weight loss effects: pancreatic lipase inhibition, bile salt hydrolase activity, stimulation of GLP-1 hormone secretion, and inhibition of preadipocyte differentiation. The OD value is then detected at a wavelength of 600 nm.

[0015] Preferably, in the method for screening probiotics for ecological coexistence, when screening probiotic combinations that promote weight loss, the OD value is specifically tested according to the following method:

[0016] The same probiotic combination was inoculated at an inoculation rate of 1%, cultured under anaerobic conditions to the logarithmic growth phase, and the OD value was measured at 600 nm.

[0017] According to another aspect of the present invention, a screening system for coexisting probiotics is also provided, which includes an OD value acquisition module and a judgment module;

[0018] The OD value acquisition module is used to acquire the OD value of the probiotic combination and submit it to the judgment module;

[0019] The judgment module is used to make judgments based on the obtained OD value according to the following principles:

[0020] If the OD value of the probiotic combination is greater than a preset threshold, it is determined that the probiotic combination is likely to coexist synergistically in vivo; otherwise, the possibility of coexisting synergistically in vivo is small. The preset threshold is between 1.9 and 2.1.

[0021] Preferably, in the screening system for coexisting probiotics, the OD value acquisition module obtains the OD value according to the following detection method:

[0022] The same probiotic combination was inoculated at a preset ratio and cultured until the logarithmic growth phase. The OD value was then measured under the corresponding wavelength conditions. The preset ratio was 1%-3%.

[0023] Preferably, the OD value acquisition module of the eco-coexisting probiotic screening system is used to acquire the OD value of a combination of probiotics with two or more functions, including pancreatic lipase inhibition, bile salt hydrolase activity, stimulation of GLP-1 hormone secretion, and inhibition of preadipocyte differentiation, when screening for probiotic combinations with weight loss effects, under in vitro co-culture conditions of 600 nm.

[0024] Preferably, in the screening system for coexisting probiotics, the OD value acquisition module obtains the OD value by means of the following detection method when screening probiotic combinations that promote weight loss:

[0025] The same probiotic combination was inoculated at an inoculation rate of 1%, cultured under anaerobic conditions to the logarithmic growth phase, and the OD value was measured at 600 nm.

[0026] Preferably, in the screening system for coexisting probiotics, the judgment module judges and screens coexisting probiotics according to the principle that if the OD value of the probiotic combination is greater than 1.9, the probiotic combination is likely to coexist synergistically in vivo, and vice versa.

[0027] In summary, compared with the prior art, the above-described technical solutions conceived by this invention can achieve the following beneficial effects:

[0028] The present invention provides a method for screening probiotics that exhibit symbiotic coexistence. This method obtains probiotic combinations through functional pairing and then selects combinations with a high probability of co-existing symbiotically in vivo based on the OD value of the probiotic combinations after in vitro co-culture exceeding a preset threshold (1.9-2.1). Compared to random pairing, functional pairing yields more effective probiotic combinations. Furthermore, using the OD threshold method based on in vitro co-culture to screen probiotic combinations that can co-exist symbiotically in vivo effectively shortens the experimental cycle and facilitates the rapid screening of probiotic combinations that possess multiple functions and exhibit symbiotic coexistence in vivo. Attached Figure Description

[0029] Figure 1This is the result of screening for strains with pancreatic lipase-inhibiting function;

[0030] Figure 1 In Figure A, the color development of the rhodamine plate is shown; in Figure B, the screening results of pancreatic lipase inhibition function of 60 bacterial strains are shown.

[0031] Figure 2 The results are the screening results of strains with bile salt hydrolase activity;

[0032] Figure 2 A shows the formation of the sedimentation zone on the plate. Left side: ordinary MRS plate; right side: BSH activity detection plate; B shows the screening results of BSH activity of 60 strains.

[0033] Figure 3 This is the result of screening for strains that stimulate GLP-1 hormone secretion;

[0034] Figure 3 A represents the initial screening of 60 bacterial strains under stimulation; B represents the repeat experiment of 13 bacterial strains with increased hormone secretion compared to the control group.

[0035] Figure 4 This is the result of OD value detection for the co-culture of probiotic combinations;

[0036] Figure 5 This is a schematic diagram of the experimental design for the abundance of the strain in vivo. Detailed Implementation

[0037] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention. Furthermore, the technical features involved in the various embodiments of this invention described below can be combined with each other as long as they do not conflict with each other.

[0038] Data from in vitro studies indicate that optimal probiotic combination regimens derived from studies of individual strains do not always reflect in vivo realities. Some probiotics may inhibit other co-administered strains and limit overall efficacy. Different microorganisms, due to their symbiotic relationships, compete for scarce nutrients and limited space, such as releasing secretions to acquire resources, losing important genes that allow them to obtain products from external sources, poisoning neighboring cells, or preventing the colonization of other microorganisms, which may affect the growth of other bacteria.

[0039] Therefore, although numerous studies on individual probiotic strains have provided useful information about potential mechanisms of action and identified effective candidate strains, further experiments are needed to determine whether they maintain their beneficial effects when combined with other probiotics. How to rationally design and combine different strains, quickly screen for combinations where there are no antagonistic interactions, and clarify the effects of probiotic combinations to enable them to exert a greater and more effective effect is an important direction for future research.

[0040] This invention provides a method for screening probiotics that coexist in an ecological manner, comprising the following steps:

[0041] Based on high-throughput screening targeting multiple targets, single strains with preset high-throughput capabilities were obtained. These high-throughput strains were then combined according to their functional combinations, co-cultured in vitro, and their OD values ​​were measured. Screening was then performed according to the following principles:

[0042] If the OD value of the probiotic combination is greater than a preset threshold, it is determined that the probiotic combination is likely to coexist synergistically; otherwise, the possibility of synergistic coexistence is small. The preset threshold is between 1.9 and 2.1.

[0043] The OD value is specifically measured as follows:

[0044] The same probiotic combination was inoculated at a preset ratio and cultured until the logarithmic growth phase. The OD value was then measured under the corresponding wavelength conditions. The preset ratio was 1%-3%.

[0045] The OD value refers to the energy difference of light before and after passing through the analyte, i.e., the energy absorbed by the analyte. At a specific wavelength, there is a quantitative relationship between the concentration of the same analyte and the absorbed energy, and the concentration of the analyte can be determined by the OD value. Experimental statistics show that the OD value of most strains is between 1.5 and 1.8, and the OD value of a single strain with good growth is about 1.8. Therefore, this OD value can only indicate that at least one strain in the combination is growing well, but cannot characterize the occurrence of synergistic effects. However, experiments have found that probiotic combinations with OD values ​​above 1.9 in in vitro co-culture have a good probability of symbiosis in vivo and are more likely to form a synergistic and balanced ecological environment. Thus, a simpler in vitro screening experiment can replace in vivo application evaluation, improving the accuracy of the evaluation of strain combinations, achieving high-throughput screening, and reducing the blindness of in vitro and in vivo experiments. This is of great significance in terms of theoretical guidance, reducing the development cost of probiotic combinations, and shortening the development cycle.

[0046] Experiments show that, during the screening of probiotic combinations for weight loss, the probiotic combinations obtained in the in vivo symbiotic verification experiment all had OD values ​​above 1.9. The OD value is the OD value detected at a wavelength of 600nm, specifically obtained according to the following testing methods:

[0047] The same probiotic combination was inoculated at an inoculation rate of 1%, cultured under anaerobic conditions to the logarithmic growth phase, and the OD value was measured at 600 nm.

[0048] When screening probiotic combinations for weight loss, the preset functions include four functions: inhibition of pancreatic lipase, bile salt hydrolase activity, stimulation of GLP-1 hormone secretion, and inhibition of preadipocyte differentiation; the high-throughput screening is conducted according to the following principles:

[0049] (1) Screening of strains with pancreatic lipase inhibitory function: The Rhodane chromogenic plate method was used, and the diameter of the aperture was less than 15 mm as the evaluation criterion for the inhibitory effect. Strains with significant inhibitory effects were selected.

[0050] (2) Screening of strains with bile salt hydrolase activity: The plate method was used for qualitative detection, and the diameter of the precipitate ring was greater than 12 mm as the evaluation criterion for BSH activity. Strains with significant activity were selected.

[0051] (3) Screening of strains that stimulate GLP-1 hormone secretion: Cell models were used for screening to select strains that secrete more GLP-1 hormone than the control group, and strains that significantly increased the secretion of GLP-1 hormone were preferred.

[0052] (4) Screening of strains that inhibit preadipocyte differentiation: Using the 3T3-L1 preadipocyte model, the inhibitory effect of the strains on cell differentiation was judged by Oil Red staining based on the formation of lipid droplets. Compared with the control group, the screening was based on the significant reduction of the proportion of colored lipid droplets as the evaluation criterion for inhibitory effect, and strains with significant inhibitory effect were selected.

[0053] The different strains obtained are combined according to their functional combinations, specifically:

[0054] Probiotic combinations should be made that possess two or more of the above-mentioned functions, preferably with four functions, and the number of strains should generally not exceed four.

[0055] The same probiotic combination was inoculated at an inoculation rate of 1%, cultured under anaerobic conditions to the logarithmic growth phase, and the OD value was measured at 600 nm. Probiotic combinations with an OD value of 1.9 or higher were screened.

[0056] Furthermore, based on in vitro co-culture-functional verification, combinations with two or more functions and good effects are screened, with combinations having four functions being preferred; in vivo colonization experiments can be used to verify the effects of probiotic combinations in animal models.

[0057] In addition, the present invention also provides a screening system for ecologically coexisting probiotics, which includes an OD value acquisition module and a judgment module;

[0058] The OD value acquisition module is used to acquire the OD value of the probiotic combination and submit it to the judgment module;

[0059] The judgment module is used to make judgments based on the obtained OD value according to the following principles:

[0060] If the OD value of the probiotic combination is greater than a preset threshold, it is determined that the probiotic combination is likely to coexist synergistically in vivo; otherwise, the possibility of coexisting synergistically in vivo is small. The preset threshold is between 1.9 and 2.1.

[0061] Furthermore, the OD value acquisition module obtains the OD value according to the following detection method:

[0062] The same probiotic combination was inoculated at a preset ratio and cultured until the logarithmic growth phase. The OD value was then measured under the corresponding wavelength conditions. The preset ratio was 1%-3%.

[0063] The OD value acquisition module, when screening probiotic combinations for weight loss, is used to obtain the OD value of probiotic combinations with two or more functions, including pancreatic lipase inhibition, bile salt hydrolase activity, stimulation of GLP-1 hormone secretion, and inhibition of preadipocyte differentiation, after in vitro co-culturing at 600 nm. The OD value is preferably obtained according to the following detection method:

[0064] The same probiotic combination was inoculated at an inoculation rate of 1%, cultured under anaerobic conditions to the logarithmic growth phase, and the OD value was measured at 600 nm.

[0065] The judgment module judges and screens probiotics based on the principle that if the OD value of the probiotic combination is greater than 1.9, the probiotic combination is likely to coexist synergistically in vivo, and vice versa.

[0066] The following is an example:

[0067] Example 1

[0068] The bacterial strains used in the experiment are shown in the table below (Table 1):

[0069] Table 1. Strains used in the experiment

[0070]

[0071]

[0072] 1.1 Screening of functional strains

[0073] (1) Screening of strains with pancreatic lipase inhibitory function

[0074] The Rodin definite color plate method was used for screening;

[0075] like Figure 1 As shown in Figure A, under ultraviolet light irradiation, a smaller aperture diameter indicates a better inhibitory effect of the strain on pancreatic lipase. Compared with the control group, the aperture diameter of the strain was significantly smaller, indicating a better inhibitory effect on pancreatic lipase activity. In this experiment, a plate colorimetric experiment was performed on the supernatant of 60 bacterial strains, with an aperture diameter less than 15 mm used as the evaluation criterion for inhibitory effect. The results are as follows... Figure 1 As shown in B, a total of 6 positive strains were obtained through screening.

[0076] (2) Screening of BSH-active strains

[0077] BSH activity was screened using the direct plate method;

[0078] BSH activity assay plates: Add 0.5% sodium deoxytaurocholate and 1.5% agar to MRS medium and sterilize at 121℃ for 15 min. If preparing a control plate, omit the sodium deoxytaurocholate. After sterilization, pour the mixture into sterile plates. After the MRS medium cools and solidifies, invert the plates and place them in an anaerobic operating room for 48 h for deoxygenation. Before the experiment, carefully place sterile filter paper discs on the MRS culture plates, add 5 μl of bacterial suspension to each disc, and incubate the plates anaerobically at 37℃ for 48 h. If a white precipitate forms around the filter paper disc, it indicates that the strain has BSH activity; the larger the diameter of the precipitate ring, the stronger the enzyme activity. The absence of precipitate indicates a negative result.

[0079] BSH can hydrolyze sodium deoxytaurocholate added to a petri dish, thus forming a precipitate ring on the plate. The size of the precipitate ring can indicate the activity of BSH. Figure 2 As shown in Figure A, the left side is a standard MRS plate, and the right side is a BSH detection plate supplemented with sodium deoxytaurocholate. It can be seen that all four bacterial strains on the right side formed distinct precipitation zones on the plate, indicating that strains 41-44 possess BSH activity. This study tested 60 candidate bacteria, using a precipitation zone diameter greater than 12 mm as the evaluation criterion for BSH activity. The results are as follows... Figure 2 As shown in B, a total of 8 positive strains were obtained through screening.

[0080] (3) Screening of strains that stimulate GLP-1 hormone secretion

[0081] The cell model (intestinal secretory cells STC-1) was used to screen strains that stimulate GLP-1 hormone secretion;

[0082] 48 hours before the experiment, STC-1 cells were cultured at a density of 2 × 10⁶ cells per well. 5Cells were seeded at a density of 100% in 24-well plates and cultured to 80% confluence. Before bacterial stimulation, cells were gently washed twice with phosphate-buffered saline and starved for 30 min in glucose- and L-glutamine-free DMEM. The probiotics were washed three times with PBS, and bacterial particles were collected by centrifugation at 7000 rpm for 5 min. The suspension was then adjusted to a concentration of 10⁻¹⁰ with glucose- and L-glutamine-free DMEM. 7 Add CFU to the well. After incubation for 2 hours, collect the supernatant into a 1.5 mL centrifuge tube, centrifuge at 1000×g at 4℃ for 20 min to remove cells and bacterial debris, and freeze at -20℃ for later testing.

[0083] GLP-1 content determination: Performed according to the instructions of the mouse GLP-1 ELISA kit (Elabscience, China);

[0084] Preliminary screening of 60 bacterial strains yielded the following results: Figure 3 As shown in Figure A, some strains were found to indeed stimulate the secretion of GLP-1 hormone. The experiment was repeated with 13 strains that showed an increase in hormone secretion compared to the control group, and the results showed that ( Figure 3 B) Eleven strains of bacteria were indeed able to significantly stimulate the secretion of GLP-1 hormone.

[0085] (4) Screening of strains that inhibit preadipocyte differentiation

[0086] This part of the experiment used 3T3-1 cells (mouse embryonic fibroblasts), which can differentiate into adipocytes under the induction of an inducer. During the induction of cell differentiation with the inducer, bacteria were added at a ratio of MOI=10 for intervention. Successfully differentiated adipocytes secrete a large number of lipid droplets. Oil Red staining was used to determine the inhibitory effect of the bacteria on cell differentiation based on the formation of lipid droplets. Compared with the control group, the differentiation of cells treated with b15m2 was significantly inhibited, and the number of stained lipid droplets was significantly reduced.

[0087] This experiment was conducted on 60 strains of bacteria. After the staining results were photographed, Image-Pro Plus 6.0 was used to quantitatively analyze the proportion of the red area to the total area. After statistical analysis, 10 positive strains were screened out from the strains with significant differences.

[0088] In summary, the experimental results show that 28 functional strains were screened out from 60 candidate bacteria. Most of these bacteria have only one function, and only 7 have two functions.

[0089] 1.2 Screening of Probiotic Combinations

[0090] (1) Strains according to functional combinations

[0091] Combinations were designed based on the condition that all four functions are included, and the number of strains in each combination should not exceed four, resulting in a total of 653 combinations.

[0092] (2) In vitro co-culture screening

[0093] The obtained 653 combinations were subjected to in vitro co-culture experiments. After each strain was cultured individually for 48 hours, the same probiotic combination was inoculated into the same 5mL vial at a 1% inoculum size and cultured anaerobically at 37℃ for 12 hours. The OD value was measured after 12 hours of co-culture. All combinations were then co-cultured at 37℃ for 12 hours, and the OD value was measured at 600nm. Figure 4 As shown;

[0094] Depend on Figure 4 It can be seen that the OD values ​​of most combinations are between 1.5 and 1.8. However, this range of OD values ​​only indicates that at least one strain in the combination is growing well, and does not rule out the possibility that the growth of other strains is inhibited, nor does it indicate that all four functions in the combination are effective.

[0095] This experiment conducted in vivo colonization experiments on various probiotic combinations. A schematic diagram of the in vivo abundance experiment flowchart for probiotic combinations is shown below. Figure 5 As shown, the results revealed that 63 probiotic combinations with OD values ​​above 1.9 were more likely to coexist in vivo.

[0096] Example 2: In vivo colonization verification of a probiotic combination with four functions

[0097] (1) After conducting four different functional verification experiments on 63 combinations selected from the co-culture experiment, four combinations possessed all the functional effects, as shown in Table 2.

[0098] Table 2 Results of combined in vitro screening

[0099]

[0100]

[0101] (2) In vivo colonization experiment of probiotic combination

[0102] PCR was used to detect the colonization of bacterial strains in vivo. Using fecal genomic DNA at different time points as a template, the presence of bacteria in feces was detected by PCR.

[0103] Combining the results of in vitro co-culture-functional verification and in vivo experiments, it was found that among the four combinations obtained in vitro, the bacteria contained in three combinations could colonize mice for more than 3 days. The bacterial strains contained in the combinations are shown in Table 3.

[0104] Table 3. Combination of in vitro and in vivo experiments for screening

[0105]

[0106] Those skilled in the art will readily understand that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for screening probiotics that coexist ecologically, characterized in that, Includes the following steps: Based on high-throughput screening of multiple targets, a combination of probiotics with screening effects on weight loss was obtained, according to single strains with high-throughput effects such as pancreatic lipase inhibition, bile salt hydrolase activity, stimulation of GLP-1 hormone secretion, and inhibition of preadipocyte differentiation. Different high-throughput bacterial strains are combined according to their functional combinations to obtain probiotic combinations; the high-throughput bacterial strains are selected from Bifidobacterium adolescentis, Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium animalis, Bifidobacterium bifidum, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus salivarius, and / or Lactococcus lactis. We screened probiotic combinations that promote weight loss, selecting combinations based on four functions: pancreatic lipase inhibition, bile salt hydrolase activity, stimulation of GLP-1 hormone secretion, and inhibition of preadipocyte differentiation. The number of probiotic combinations was no more than four. In vitro co-culture was performed, and the OD value of the probiotic combination was measured. In vitro experiments were used to replace in vivo evaluation, and the selection was based on the following principles: If the OD value of the probiotic combination is greater than a preset threshold, it is determined that the probiotic combination is likely to coexist synergistically in vivo; otherwise, the possibility of coexisting synergistically in vivo is small. The preset threshold is between 1.9 and 2.

1. The OD value is specifically measured as follows: The same probiotic combination was cultured under anaerobic conditions to the logarithmic growth phase, and the OD value was measured at 600 nm.

2. The screening method for eco-coexisting probiotics as described in claim 1, characterized in that, The OD value is specifically measured as follows: The same probiotic combination was inoculated at a preset ratio and cultured until the logarithmic growth phase. The OD value was then measured under the corresponding wavelength conditions. The preset ratio was 1%-3%.

3. A screening system for ecologically coexisting probiotics, characterized in that, Includes an OD value acquisition module and a judgment module; The OD value acquisition module is used to acquire the OD value of the probiotic combination and submit it to the judgment module; the probiotic combination is selected according to four functions: pancreatic lipase inhibition, bile salt hydrolase activity, stimulation of GLP-1 hormone secretion, and inhibition of preadipocyte differentiation, and the number of combinations is no more than four strains; the probiotics are selected from Bifidobacterium adolescentis, Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium animalis, Bifidobacterium bifidum, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus salivarius, and / or Lactococcus lactis; The OD value is specifically measured as follows: The same probiotic combination was cultured under anaerobic conditions to the logarithmic growth phase, and the OD value was measured at 600 nm. The judgment module is used to evaluate in vitro applications based on the obtained OD value, specifically according to the following principles: If the OD value of the probiotic combination is greater than a preset threshold, it is determined that the probiotic combination is likely to coexist synergistically in vivo; otherwise, the possibility of coexisting synergistically in vivo is small. The preset threshold is between 1.9 and 2.

1.

4. The screening system for ecologically coexisting probiotics as described in claim 3, characterized in that, The OD value acquisition module obtains the OD value according to the following detection method: The same probiotic combination was inoculated at a preset ratio and cultured until the logarithmic growth phase. The OD value was then measured under the corresponding wavelength conditions. The preset ratio was 1%-3%.

5. The screening system for ecologically coexisting probiotics as described in claim 4, characterized in that, The OD value acquisition module obtains the OD value by screening probiotic combinations that promote weight loss, according to the following detection method: The same probiotic combination was inoculated at a rate of 1% and cultured under anaerobic conditions until the logarithmic growth phase.

6. The screening system for ecologically coexisting probiotics as described in claim 5, characterized in that, The judgment module judges and screens probiotics based on the principle that if the OD value of the probiotic combination is greater than 1.9, the probiotic combination is likely to coexist synergistically in vivo, and vice versa.

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