Intestinal content separation and recovery device, method and extract

Abstract

The application provides an intestinal content separation and recovery device and method, the separation and recovery device comprising a foreign matter removal module and a target content collection module; the foreign matter removal module removes the impurity part of the intestinal content, and the foreign matter removal module has a cross-flow filtration structure; and the target content collection module is mainly composed of a microporous membrane cross-flow filter. The method removes the impurities in the intestinal content in a way of coarse filtration and cross-flow fine filtration, and then obtains a concentrated liquid by using a microporous membrane cross-flow filter to collect the target content. The microporous membrane has a sponge-like porous structure, and is matched with cross-flow filtration to quickly and effectively realize the separation and recovery of the flora in the intestinal content. The separated and recovered flora dry powder accounts for at least 10%-20% of the initial wet intestinal content, and the bacterial content is at least 10 11 bacteria / 1 g of dry powder.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medical applications, and particularly relates to an intestinal content separation and recovery device, a separation and recovery method and an extract thereof. BACKGROUND

[0002] The intestinal flora of mammals is an important metabolic and immune "organ" in the body of mammals, and affects the overall metabolism of the host. The metabolism of the host is affected by both the genes of the host and the genes of the intestinal flora, and there is a "co-metabolic" process between the host and the flora. Intestinal bacteria can participate in the metabolism of the host and form a co-metabolic relationship with the host. Under normal circumstances, the intestinal flora in the body of mammals is relatively constant. Once the intestinal flora is disordered, the function of the immune system of the host will be disturbed, leading to diseases.

[0003] There are a large number of symbiotic microorganisms in the human body, most of which inhabit the human intestine, and the number exceeds 100 trillion (10 14 The intestinal microorganisms are more than 99% bacteria, including most anaerobes and a small amount of aerobic bacteria. The intestinal microorganisms are important participants in human metabolism, providing substrates, enzymes and energy for the metabolic process of humans; at the same time, the metabolites such as fatty acids promote the growth and differentiation of human epithelial cells, and participate in the synthesis of vitamins and the absorption of various ions. Since the intestine is the largest immune organ in the human body, the exchange between the intestinal microorganisms and the host on the surface of the intestinal mucosa promotes the establishment and development of the immune system, and becomes an important immune barrier of the human body. In addition, the intestinal microorganisms also provide protection for the human body by forming a "bacterial membrane barrier".

[0004] The intestinal microorganisms of the human body accompany the growth of humans at all times, not only helping to digest food, but also being a barrier for the human body to effectively resist the invasion of pathogenic bacteria from the outside world. The balanced and stable intestinal microecological environment formed in the long evolution process plays a crucial role in the nutrition, metabolism and immunity of the human body. In the long evolution process, the intestinal microorganisms have reached a good cooperation with humans, and play a crucial role in the nutrition, metabolism and immunity of the human body.

[0005] The number of various bacteria in the intestine needs to reach a relative balance to play a role. This balance is very fragile. If there is abdominal pain and diarrhea, it is mostly that the balance of the intestinal flora is destroyed. Malnutrition, obesity, diabetes and the like can be related to the imbalance of the intestinal flora. The latest research shows that Alzheimer's disease and autism can also be related to the imbalance of the intestinal flora.

[0006] In December 2013, Science magazine published the top ten scientific progress, the relationship between gut flora and human health research was included. The intestinal microflora is very complex, the intestinal flora combined in a certain proportion, mutual restraint, mutual dependence, the intestinal flora in the number of symbiotic win-win ecological balance with the host. Through diet and supplement of probiotics, can have a certain effect or influence on intestinal microflora. However, improving the dietary structure can achieve the purpose of improving the intestinal microecological balance, but the process is slow and time-consuming; compared with the variety of intestinal flora, the limited number and variety of probiotics are difficult to restore the overall balance of intestinal microecology, and the curative effect is limited. In recent years, as an important new technology in the field of medicine, fecal microbiota transplantation (FMT) has gradually been recognized by people and has been concerned by the clinic.

[0007] The feces of mammals such as humans mainly include undigested residues (such as food fiber), intestinal secretions (including mucus), proteins, white blood cells, epithelial cells, bacteria, inorganic matter (mainly calcium and phosphate), food that cannot be digested or absorbed, and water. A normal adult usually defecates once a day, and the amount of feces will change with the type of food, food intake, and the functional status of the digestive organs. For example, those who mainly eat fine grains and meat have fine and small amounts of feces; eating coarse grains, especially a large amount of vegetables, because of the high fiber content, the feces are coarse and large in amount.

[0008] There are a lot of bacteria in feces, accounting for 1 / 3 of the dry weight, most of which belong to normal flora. The main bacteria in the feces of healthy infants and young children are Bifidobacterium, Bacteroides, Enterobacter, Enterococcus, Staphylococcus, etc. In adult feces, Escherichia coli, anaerobes and enterococci are the main flora, accounting for about 80%; gas-producing bacteria, Proteus, Pseudomonas aeruginosa, etc. are mainly passing bacteria, not more than 10%. The total amount of spore bacteria (such as Clostridium) and yeast is not more than 10%. The amount and spectrum of bacteria in feces are relatively stable, and maintain ecological balance with the host (Clinical Laboratory Diagnosis and Clinical Application, Wang Ruihui, ed., Changchun: Jilin Science and Technology Press, 2017.03).

[0009] FMT is to transfer the flora (intestinal flora) in the feces of healthy people to patients, so that patients obtain the intestinal flora of healthy people, regulate intestinal flora imbalance, and rebuild the intestinal microecosystem with normal function, which provides help for the treatment of intestinal and extra-intestinal diseases. In the treatment of diseases by FMT in modern human, the state of fecal bacteria used is mostly fresh fecal bacteria liquid, or fecal clear, or fermented fecal liquid, or child fecal liquid. The "Huanglong Decoction" called by the ancient Chinese doctors is a medicine containing human fecal juice. FMT using fecal bacteria liquid has a more important problem of how to complete the separation and preservation of fecal bacteria within the time that does not affect the function of the flora, in addition to the great psychological challenge that medical staff and patients have to bear.

[0010] There are at least 1000-1150 kinds of bacteria in the human intestine, and there are about 160 dominant bacterial species in each host. Although the main composition groups of human intestinal microorganisms are very similar, there are great differences in the relative content and strain species of different microbial groups among different host individuals. The factors affecting the difference of microbial flora include the region, age, physiological condition, dietary habits of the host and other factors. Although human fecal bacteria is the only organ that can be truly shared by humans, there is no need to consider the problem of immune rejection. However, the ecological balance of the intestinal flora of each individual is indeed different, and there is even a large difference. This also increases the difficulty of FMT to some extent.

[0011] One of the keys of FMT is the preparation of the fecal bacteria to be transplanted, and the current one is fecal bacteria liquid. However, the preparation of fecal bacteria liquid has not formed a consensus, and many factors will affect the prepared fecal bacteria liquid. First, the amount of feces, because the weight of feces is not proportional to the number of flora, and there is a large individual difference, there is no unified standard to determine the amount of feces in the preparation of fecal bacteria liquid. Second, the preparation time, the fecal flora contains a lot of anaerobic bacteria, and the metabolic time of each bacterial species is different. Therefore, it is necessary to complete the separation and extraction within the shortest possible time, otherwise it is easy to cause the death of anaerobic bacteria and the great change of the relative abundance of bacterial species, which affects the clinical treatment effect. Third, the existing preparation process and method, mainly using manual coarse filtration plus centrifugal enrichment method, or microfiltration plus centrifugal enrichment method. The fecal bacteria obtained by the existing preparation method has great difference in species and abundance compared with the flora distribution in the initial intestinal contents, and the repeatability is poor. Fourth, the state of fecal bacteria liquid, there are fresh fecal bacteria liquid and freeze-dried fecal bacteria liquid; freeze-dried fecal bacteria liquid is prepared by adding glycerol to fresh fecal bacteria liquid and freezing at low temperature, or it can also be made into frozen fecal bacteria capsules.

[0012] At present, for the fecal bacteria automatic separation and recovery system, multi-stage filter is often used to remove other impurities, and some domestic documents have been disclosed, such as CN103330961B, CN105624024B and CN204097475U. However, these fecal bacteria separation and recovery systems only filter and separate the large particle impurities in the fecal solution, such as undigested food residues, to obtain a bacteria liquid. The bacteria liquid should contain a large amount of food residues, bacterial metabolites and water. The common method is to centrifuge the mixed liquid filtered by the centrifuge to separate the layers, and to discard the supernatant or the sinking material to retain the useful bacteria. Most of the intestinal bacteria are between 0.1 and 50 μm in size, and discarding the supernatant will lose a large amount of small size microorganisms. Retaining the supernatant and discarding the sinking material will lose large size microorganisms. Furthermore, due to the diversity and complexity of the composition of the fecal sample and the difference in operation of each person, the method has great difference, the result is uncontrollable, and it is impossible to realize the standardized and repeatable operation, and the operation is very inconvenient.

[0013] In addition, these methods cannot selectively screen specific intestinal microorganisms due to poor controllability, and cannot meet the future precise treatment of intestinal diseases. At the same time, the traditional filter screen filtering method is used in such devices to remove small size impurities. Since the mesh size of the fine filter screen is between 0.01 mm and 1 mm, it is easy to be blocked by impurities, thereby greatly reducing the filtering efficiency. And because the feces contain a lot of sticky ingredients, the efficiency of the traditional filter screen filtering method is reduced, and it is impossible to complete the bacteria separation and extraction within an effective time. The fecal bacteria contain a high amount of bacteria, and the existing multi-stage filtering patent technology has a low amount of bacteria per unit volume, poor repeatability of operation, and poor consistency of bacteria and sample, which increases the difficulty of bacteria transplantation.

[0014] WO2012122478A1 discloses a composition of a human fecal extract, by making the blended sample pass through a screen with a mesh size of not more than 2.0 mm, the filtrate can be collected in a centrifuge tube, and centrifuged for 10 minutes. The supernatant is removed, and the cells are resuspended in a diluent, and centrifuged again for 10 minutes. The final supernatant is discarded and the cells are resuspended in an aqueous solution to obtain the composition. The method of this document results in a large loss of bacteria, and the operation in the air will cause a large number of deaths of anaerobic bacteria.

[0015] WO2016065777A1 discloses a device for separating microorganisms in the contents of a cavity, wherein the outlet of a raw material tank is connected to the inlet of a fractional filtering device, the fractional filtering device is composed of one or more fractional filtering bottles, the outlet of a liquid storage tank is connected to the inlet of the raw material tank and at least one fractional filtering bottle, the material inlet of a closed separation material tank is connected to the material outlet of the fractional filtering device, the bacteria liquid inlet of the first fractional filtering bottle of a fractional device composed of at least one fractional bottle is connected to the bacteria liquid outlet of the separation material tank, the odor inlet of a deodorizing device is connected to the odor outlets of the raw material tank and the separation material tank through an odor pipeline, at least one air pump is arranged on the odor pipeline to guide the odors in the raw material tank and the separation material tank into the deodorizing device respectively; the separation device helps filtering through the shaking mode of shaking, shaking or fluctuation of the liquid in the fractional filtering bottle.

[0016] WO2016201114A1 discloses a method for manufacturing an oral microbiota restoration therapy (MRT) composition, the method comprising: collecting a fecal sample; purifying the fecal sample to form a purified intermediate, wherein purifying the fecal sample comprises: adding a diluent to the fecal sample; mixing the fecal sample with the diluent to form a mixture; filtering the mixture; transferring a filtrate from the filtering step to a centrifuge tube; and centrifuging the filtrate to obtain the purified intermediate; lyophilizing the purified intermediate to form a plurality of lyophilized pellets; and encapsulating the plurality of lyophilized pellets in one or more capsules.

[0017] CN201810852561.2 discloses an intestinal content separation and recovery device, method and product thereof, which uses a microporous membrane separation module combining side wall filtration and rotary separation technology to extract intestinal microorganisms, replacing traditional centrifugal separation technology. The filtration characteristics of nuclear pore membranes are utilized in the separation of intestinal contents. Different types of intestinal contents are collected by filtering through nuclear pore membranes with different pore sizes, and the products obtained are intercepted on the nuclear pore membranes. As the previous invention of the inventor, the present invention has greatly improved the acquisition of intestinal microorganisms compared to the centrifugal operation. However, since the impurity removal stage still uses conventional filtration methods, the final collection of intestinal microorganisms uses nuclear pore membranes, which are membrane structures with uniform pore sizes. Negative pressure needs to be applied to the lower end to implement cross-flow filtration. The overall filtration efficiency and the quality of the obtained intestinal microorganisms still need to be improved.

[0018] The existing extraction and treatment of fecal flora mostly use centrifugal process. However, during the centrifugal operation process, due to the differences between the intestinal contents, including various food residues, the composition and abundance of bacteria in the feces, etc., the results of each layering are different, and the lost bacterial species are different. Even if the centrifugal process parameters such as rotation speed, solid-liquid ratio, etc. are the same, the quality of the obtained bacterial flora product is also uncontrollable, and effective repeatability cannot be achieved. Not to mention that manual operation is involved in the operation process, and the repeatability of the whole process faces great challenges.

[0019] Secondly, the extraction efficiency is generally low, and the intestinal bacteria are severely lost during the extraction process, resulting in a limited number of extracted bacteria, thereby reducing the FMT treatment effect and increasing the cost of FMT.

[0020] At the same time, the fecal flora needs to be processed through multiple processes. Since the fecal flora is mainly anaerobic bacteria, centrifugal operation needs to be carried out in a nitrogen bioengineering kitchen, which becomes difficult to operate. Since not all operations can be carried out in a nitrogen bioengineering kitchen, the quality is difficult to guarantee, and the curative effect is also unstable.

[0021] Therefore, the lack of standardized fecal bacteria separation and transplantation technology in the prior art is one of the key reasons restricting the development of fecal bacteria transplantation. How to quickly and efficiently realize fecal bacteria separation and guarantee its repeatability, standardization, and consistency between the obtained fecal bacteria and the initial sample is of great significance for fecal bacteria separation. The technical field needs an intestinal content separation and recovery device and a method for separating and recovering intestinal contents, which effectively solves the problems of the above-mentioned centrifugal separation and eliminates or alleviates all or part of the defects in the prior art. SUMMARY

[0022] In view of the problems in the prior art, the purpose of the present application is first to provide an intestinal content separation and recovery device which can realize efficient, accurate and controllable collection of specific intestinal flora by adopting a micro-porous membrane ultra-fine module cross-flow filtration mode.

[0023] At the same time, the present application also provides a method for separating and recovering intestinal contents, and an extract of the separated and recovered intestinal contents.

[0024] It is emphasized that, unless otherwise specified, the terms used herein are consistent with the usual meanings of various scientific terms in the art, the meanings of professional terms defined in various technical dictionaries, textbooks, etc.

[0025] The present application provides an intestinal content separation and recovery device, wherein the device comprises a contaminant removal module and a target content collection module; the contaminant removal module removes the contaminant part of the intestinal content, and has a cross-flow filtration structure; the target content collection module is mainly composed of a micro-porous membrane cross-flow filter, the micro-porous membrane has a sponge-like porous structure, the pore size of the micro-porous membrane ranges from 0.01 to 20 μm, preferably from 0.1 to 10 μm, and more preferably from 0.2 to 5 μm, the opening rate of the micro-porous membrane is more than 30%, preferably more than 50% or 60%, and more preferably more than 70%, 80%, 85% or 90%, and the micro-porous membrane is a hydrophilic membrane.

[0026] Further, the micro-porous membrane cross-flow filter has a disc structure, and the ratio of the effective filtration area of the cross-flow disc to the overall area of the cross-flow disc is 0.3-0.9; preferably, the ratio is 0.4-0.8, and more preferably, the ratio is 0.5-0.6. The ratio represents the effective use rate of the micro-porous membrane in cross-flow filtration. Optionally, the overall area of the micro-porous membrane cross-flow disc is about 270 cm 2 , and the effective filtration area is about 150 cm 2 .

[0027] Further, the flow channel of the micro-porous membrane cross-flow filter is a spiral flow channel. The spiral cross-flow flow channel can provide effective support for the flow of liquid and utilize the micro-porous membrane for cross-flow filtration as much as possible.

[0028] Further, the cross-sectional area of the flow channel of the micro-porous membrane cross-flow filter is smaller than that of the inlet pipe, and the ratio of the former to the latter is 0.8, 0.6, or 0.5, or smaller. For example, the cross-sectional area of the inlet pipe is about 35 mm 2 , and the cross-sectional area of the flow channel of the micro-porous membrane cross-flow filter is about 15-20 mm 2 . In the present application, cross-flow filtration can be achieved without using a pressurizing or negative pressure device, but this does not mean that the corresponding device cannot be used, which will increase the additional cost.

[0029] Further, the separation and recovery device has a pump for conveying the intestinal content solution from the contaminant removal module to the target content collection module through a pipeline. Preferably, the pump is a peristaltic pump to avoid cross-infection of the intestinal content solution and meet the requirements of medical consumables.

[0030] Further, the contaminant removal module comprises multiple stages of filtration, such as coarse filtration and fine filtration modules. Preferably, the fine filtration module adopts cross-flow filtration.

[0031] Further, the concentrated solution of cross-flow filtration is mixed with the permeate of the previous stage of filtration to form a circulation loop.

[0032] Further, the microporous membrane material includes one or more of polyethersulfone membrane, cellulose membrane, polymethacrylate membrane, which is modified or unmodified. Preferably, the microporous membrane polyethersulfone membrane is a wet casting phase inversion membrane. The polyethersulfone membrane is optionally a modified polyethersulfone membrane, which has a disc filter membrane 003 (0.03 μm), 010 (0.1 μm), 020 (0.2 μm), 045 (0.45 μm), 065 (0.65 μm), 080 (0.80 μm), 120 (1.2 μm), 500 (5.0 μm), 1000 (10 μm). The microporous membrane of the present application has a network structure formed by connecting particles with belt-like fibers, and a non-uniform pore structure presents a three-dimensional or three-dimensional pore variation, a sponge-like porous structure, and has a very high porosity. Such a membrane structure exhibits excellent performance in cross-flow filtration relative to a uniform nuclear pore membrane, without the need for special use of pressurized or negative pressure equipment, realizing fast and efficient cross-flow filtration. At the same time, in order to avoid the rupture of the microporous membrane during use, a gasket can be added to the back of the microporous membrane.

[0033] Further, according to an embodiment of the present application, an intestinal content separation and recovery device is provided, wherein the intestinal content separation and recovery device comprises a foreign matter removal module and a target content collection module, the foreign matter removal module comprising a stirring coarse filtration module and a fine filtration module;

[0034] The stirring coarse filtration module is a container with a stirring device at the upper part, a coarse filtration screen is arranged at the lower part of the container, and a liquid outlet and a reflux port are arranged at the bottom of the container;

[0035] The fine filtration module comprises a fine cross-flow disc filter;

[0036] After the intestinal content (feces) is mixed with a solvent (such as normal saline), the intestinal content is fully stirred by the stirring device to make the intestinal content fully dispersed in the solvent, then the larger size impurities are removed by the coarse filtration screen, and then the intestinal content is transported to the fine cross-flow disc filter by the first conveying pump, and the finer impurities are removed by the fine cross-flow disc filter;

[0037] The target content collection module comprises a microporous membrane cross-flow disc filter with a microporous membrane screen, the permeate filtered by the fine filtration module is transported to the microporous membrane cross-flow disc filter by the second conveying pump, and the target content is collected in the concentrated liquid of the microporous membrane cross-flow disc filter after cross-flow filtration through the microporous membrane screen.

[0038] Further, the pore size of the filter screen of the fine cross-flow disc filter is 5-100 μm, preferably 5-50 μm, and preferably 10 μm, 20 μm, 25 μm or 30 μm.

[0039] Further, the filter screen (microporous membrane) of the microfiltration cross-flow disc filter has a pore size of 0.01-20 μm, preferably 0.1-10 μm, preferably 0.2 μm, 0.5 μm, 1 μm, 2 μm, 5 μm or 7 μm.

[0040] Further, the fine filtration module further comprises a liquid storage box, the permeate filtered by the fine filtration cross-flow disc filter enters the liquid storage box, and then enters the target content collection module. The concentrated liquid of the fine filtration is returned to the container through the pipeline through the backflow port, mixed with the permeate after the coarse filtration, and forms a circulating loop.

[0041] Further, the target content collection module further comprises a permeate waste liquid box, the permeate filtered by the microfiltration cross-flow disc filter enters the permeate waste liquid box, and the concentrated liquid is returned to the liquid storage box of the fine filtration module through the pipeline, and the target content is collected in the concentrated liquid of the microfiltration cross-flow disc filter after circulating filtration for 10 to 60 minutes, preferably 50 minutes.

[0042] Further, the target content collection module comprises one microfiltration cross-flow disc filter; or the target content collection module comprises a plurality of microfiltration cross-flow disc filters with different pore sizes, and the microfiltration cross-flow disc filters with different pore sizes are arranged in order from large to small in terms of pore size, and are used to collect target contents of different sizes.

[0043] Further, the target content is intestinal microorganisms (intestinal flora), including Bifidobacterium, Lactobacillus, Escherichia coli, Enterococcus, etc., and the size of the intestinal microorganisms is between 0.1 μm and 50 μm.

[0044] Further, an outer shell is arranged outside the container of the stirring and coarse filtration module, a top of the outer shell is provided with an exhaust port, and odor gas generated by the feces is discharged through the exhaust port and removed by a deodorization device.

[0045] Further, the separation and recovery device is provided with an oxygen removal module to ensure and maintain a low-oxygen or oxygen-free atmosphere for the intestinal contents during the separation and recovery process.

[0046] The application also provides a separation and recovery method of intestinal contents, which is performed by using the device of the application, comprising:

[0047] Step 1) dispersion: mixing the intestinal contents with a liquid dispersion medium, the ratio of the intestinal contents to the liquid dispersion medium being 100 g:200-5000 ml, preferably 100 g:300-1000 ml, or 100 g:500-800 ml, and the obtained mixture is dispersed by a mechanical method to form a dispersion mixture;

[0048] Step 2) removing impurities: the dispersion mixture is filtered by a filter screen structure, the filtering includes cross-flow filtration, obtaining a permeate (filtrate);

[0049] Step 3) collecting: the permeate of step 2) is filtered by a micro-porous membrane cross-flow filter, obtaining a concentrated solution and a filtration waste liquid, the concentrated solution is used to collect the target content;

[0050] The micro-porous membrane of the micro-porous membrane cross-flow filter has a sponge-like porous structure, the pore size of the micro-porous membrane ranges from 0.01 to 20 μm, preferably, the pore size of the micro-porous membrane ranges from 0.1 to 10 μm, preferably, the pore size of the micro-porous membrane ranges from 0.2 to 5 μm, the opening rate of the micro-porous membrane is more than 30%, preferably 50%, 60% or more, preferably 70%, 80%, 85%, 90% or more, and the micro-porous membrane is a hydrophilic film.

[0051] Further, the content of bacteria in the dispersion mixture of step 1) is at least 10 8 cells / ml, or 10 9 cells / ml, or 10 10 cells / ml.

[0052] Further, the concentrated solution of step 3) is mixed with the permeate of step 2) to form a circulation loop.

[0053] Further, the filtering and removing impurities of step 2) are divided into coarse filtration and fine filtration, the pore size of the filter screen for coarse filtration is greater than 10 μm, or 50 μm, or 100 μm, or 200 μm, or 500 μm, obtaining a coarse permeate (filtrate), and the coarse permeate is then subjected to fine filtration.

[0054] Further, the fine filtration of step 2) is cross-flow filtration, the pore size of the filter screen for fine filtration is 5-100 μm, preferably 5-50 μm, preferably 10 μm, 20 μm, 25 μm or 30 μm, obtaining a fine permeate (filtrate) and a concentrated solution, wherein the fine permeate is subjected to filtration by a micro-porous membrane cross-flow filter in step 3), obtaining a concentrated solution and a filtration waste liquid.

[0055] Further, the concentrated solution of step 3) is returned to the previous stage and mixed with the permeate to form a circulation loop.

[0056] Further, the mechanical method of step 1) is stirring, or other effective methods for dispersing similar solid-liquid mixtures, such as ball milling, oscillation, etc.

[0057] Further, the concentrated bacterial solution of step 3) is prepared into a dry powder by freeze-drying.

[0058] A composition comprising an intestinal content extract obtained by using the intestinal content separation and recovery device or by the intestinal content separation and recovery method described above, the intestinal content extract having a size of 0.01-50 microns, or 0.1-40 microns, or 0.2-30 microns, or 0.5-20 microns.

[0059] Further, the intestinal content extract is a concentrated bacterial solution, which is freeze-dried to form a dry powder, and the bacterial content is at least 10 11 bacteria per 1g of dry powder, or 2x10 11 bacteria per 1g of dry powder, or 5x10 11 bacteria per 1g of dry powder, or 10 12 bacteria per 1g of dry powder.

[0060] The distribution (species and abundance) of the bacterial flora of the intestinal content extract is very close to the distribution of the bacterial flora in the intestinal content.

[0061] For the specific material to be processed, i.e., the intestinal content, due to its high viscosity and large individual differences, the existing bacterial flora extraction often uses centrifugal separation, but centrifugal separation causes a large amount of bacteria to be separated and discarded, and the quality controllability is poor: a large amount of bacteria is lost, the biological diversity is low, the repeatability is poor, the operation is inconvenient, the environmental control is poor, and the bacterial survival rate is low (a large amount of anaerobic bacteria in the intestinal tract), thereby causing poor or unstable curative effect.

[0062] The reproduction speeds of the bacterial species contained in the intestinal content are different, and if the separation and extraction of the intestinal content cannot be completed within an effective time, such as 1.5 hours, preferably 1 hour, the distribution of the bacterial species in the obtained extract will be different from the distribution of the bacterial species in the initial intestinal content.

[0063] The intestinal content separation and recovery device of the present application does not use a centrifugal component, does not need to use a centrifugal method to obtain the target content, but uses cross-flow filtration and a microporous membrane to obtain the target content, ensures the filtration efficiency, and completes the extraction within an effective time; solves the problems of blockage and low efficiency; can effectively extract all bacterial species and retain the relative abundance of the original bacterial species.

[0064] The present application uses a microporous membrane cross-flow filtration method to enrich the target intestinal content, instead of the traditional centrifugal separation technology, which can make the extraction process more precise and controllable, has high separation efficiency, is simple to operate, has high automation degree, and can selectively separate the required intestinal content by using microporous membranes with different pore sizes through size effect.

[0065] The present application divides the filtration into coarse filtration and fine filtration, and adopts the cross-flow filtration mode to perform the fine filtration. In the traditional fine filtration mode, a filter screen is adopted, and the fine filtration screen has a mesh size of 0.01-1 mm, which is easily blocked by impurities, thereby greatly reducing the filtration efficiency. Moreover, the traditional filter screen filtration mode has a reduced efficiency due to the fact that the feces contain a large amount of sticky components. Thus, a large amount of target microorganisms cannot smoothly pass through the fine filtration screen due to the blockage of the fine filtration screen, and the content of the target microorganisms collected finally is greatly reduced. The cross-flow filtration mode is adopted to remove the small-size impurities in the feces sample, and the tangential force parallel to the membrane surface generated by the water flow continuously flushes the retained substances on the membrane surface, prevents the small holes from being blocked, improves the separation efficiency, greatly increases the probability of the microorganisms passing through the filter screen, and prolongs the service life of the filter screen.

[0066] The traditional fine filtration module is easily blocked, and it is difficult to adopt a filter screen with a pore size of less than 20 μm to perform the filtration. Thus, a high proportion of small-size impurities is also brought into the filtrate. The fine filtration module of the present application adopts the cross-flow filtration mode, and thus the fine filtration module can select a cross-flow filter with a mesh size of 15 μm or even smaller. Thus, the content of the smaller-size impurities passing through the cross-flow filter can be greatly reduced, and the purity of the target content collected by the target content collection module is higher.

[0067] The present application adopts the micro-porous membrane cross-flow filtration mode to enrich the target intestinal content after the fine filtration step, and the fine filtration module and the target content collection module are cooperated: the target content with different sizes is enriched by the design of the filter pore size of the fine filtration module and the target content collection module. For example, the fine filtration module adopts a cross-flow filter with a pore size of 5 μm, and the target content collection module adopts a cross-flow filter with a pore size of 1 μm, and the target content with a size of 1-5 μm can be selectively collected.

[0068] The cross-flow filtration of the fine filtration module and the target content collection module of the present application preferably adopts the cross-flow disc filter, and the cross-flow filtration is performed through the pipes arranged in a spiral shape inside the micro cross-flow disc filter. The effective filtration area can be greatly increased, and thus the volume of the filter can be reduced.

[0069] The separation and recovery method of the intestinal content of the present application has a higher extraction rate and a faster extraction efficiency. The percentage of the separated and recovered bacterial population dry powder relative to the initial wet intestinal content is at least 10%-20%, and the bacterial content is at least 10 11 / 1 g of dry bacterial powder. That is, 100 g of initial wet feces can obtain at least 10 g-20 g of bacterial population dry powder. The extraction efficiency of the number of bacteria of the present application is at least 6.7×10 9 / (cm 2 ·h) to 1.34×10 10 / (cm 2• h), i.e. 100 g of initial wet feces, under an effective filtration area of about 150 cm 2 The separation and recovery can be completed in 60 min at the longest under an effective filtration area of about 150 cm 12 The obtained dry powder of the bacterial flora is at least 10 g-20 g, and the bacterial amount is 1 x 10 12 to 2 x 10 BRIEF DESCRIPTION OF DRAWINGS

[0070] The accompanying drawings, which are included to provide a further understanding of the embodiments and are incorporated in and constitute a part of this application, illustrate embodiments of the application and together with the description serve to explain the application. In the drawings:

[0071] Figure 1 is a separation and extraction system of intestinal contents of Example 1.

[0072] Figure 2 is a schematic diagram of a separation and extraction system of intestinal contents by a traditional centrifugal separation method in Comparative Example 1.

[0073] Figure 3 is a photograph under 200 times of a microscope of a product collected in a concentrated solution of a micro-porous membrane cross-flow disk filter (12) by the separation and recovery device of intestinal contents in Example 1.

[0074] Figure 4 is a photograph under 200 times of a microscope of 2 g of solid precipitate dissolved in 25 ml of normal saline from a lower precipitate obtained by the traditional centrifugal separation method in Comparative Example 1.

[0075] Figure 5 is a photograph under 200 times of a microscope of a product collected in a concentrated solution of a micro-porous membrane cross-flow disk filter by the separation and recovery device of intestinal contents in Comparative Example 2.

[0076] Figure 6 is a schematic diagram of a structure of a cross-flow disk filter of Example 1.

[0077] Figure 7 is a picture of a sample after centrifugation in Comparative Example 1 (left picture a: a picture of a sample after the first centrifugation; middle picture b: a picture of a sample after the second centrifugation; right picture c: a picture of a sample after the third centrifugation).

[0078] Figure 8 is an electron microscope photograph of supernatant after centrifugation in Comparative Example 1 (left picture a: supernatant after the first centrifugation; middle picture b: supernatant after the second centrifugation; right picture c: supernatant after the third centrifugation).

[0079] Figure 9is a plot of Alpha diversity of the target contents isolated and recovered from Example 6 and Comparative Example 3.

[0080] Figure 10 is a plot of Beta diversity of the target contents isolated and recovered from Example 6 and Comparative Example 3, raw feces.

[0081] Figure 11 is a plot of Beta diversity of the target contents isolated and recovered from Example 6 and Comparative Example 3, raw feces.

[0082] Figure 12 is a plot of species diversity of the target contents isolated and recovered from Example 6 and Comparative Example 3, raw feces.

[0083] Figure 13 is a plot of log values of average copy number / g dry powder of the target contents isolated and recovered from Example 7 and Comparative Examples 4-5.

[0084] Element Number Explanation

[0085] 1: impurity removal module; 2: target content collection module; 3: stirring coarse filtration module; 4: fine filtration module; 5: stirring device; 6: container; 7: coarse filtration filter screen; 8: liquid outlet; 9: backflow port; 10: fine filtration cross-flow disc filter; 11: first delivery pump; 12: microporous membrane cross-flow disc filter; 13: second delivery pump; 14: liquid storage box; 15: waste liquid box; 16: housing; 17: exhaust port; 18: deodorizing device;

[0086] 201: stirring coarse filtration module; 202: fine filtration module; 203: stirring device; 204: container; 205: coarse filtration filter screen; 206: fine filtration filter screen;

[0087] 501: cross-flow liquid inlet; 502: concentrated liquid outlet; 503: spiral cross-flow filtration pipe. DETAILED DESCRIPTION

[0088] In order to make the objects, technical solutions, and advantages of the present application clearer, the technical solutions of the present application will be described below in connection with specific embodiments of the present application and corresponding drawings. Obviously, the described embodiments are only some of the embodiments of the present application, rather than all the embodiments. Based on the embodiments of the present application, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the scope of protection of the present application.

[0089] In the present application, the testing of the intestinal content flora mainly adopts 16S rRNA gene amplicon sequencing.

[0090] The technical solutions provided by embodiments of the present application are described in detail below with reference to the drawings.

[0091] Embodiment 1

[0092] As shown in Figure 1 , an intestinal content separation and recovery device according to an embodiment of the present application is shown, which comprises a foreign matter separation module 1 and a target content collection module 2. The foreign matter separation module 1 comprises a stirring coarse filtration module 3 and a fine filtration module 4.

[0093] The stirring coarse filtration module 3 comprises a container 6 with a stirring device 5 in the upper part of the container 6. A coarse filtration filter screen 7 is arranged at the lower part of the container 6, and the stirring device 5 is located above the coarse filtration filter screen 7. A liquid outlet 8 and a backflow port 9 are arranged at the bottom of the container 6 and are spaced apart from each other.

[0094] In operation, for example, 130 g of initial wet intestinal content (feces) can be mixed with 750 ml of physiological saline in the container 6, and then fully stirred and dispersed by the stirring device 5. The mixture is filtered through the coarse filtration filter screen 7, and then discharged through the liquid outlet 8. The larger impurities in the feces can be removed.

[0095] The fine filtration module 4 comprises a fine filtration cross-flow disc filter 10. The liquid filtered through the coarse filtration is discharged from the liquid outlet 8 and then conveyed to the fine filtration cross-flow disc filter 10 by a first conveying pump 11. The fine filtration cross-flow disc filter 10 removes finer impurities. The fine filtration module 4 further comprises a liquid storage box 14 in fluid communication with the fine filtration cross-flow disc filter 10. The permeate filtered through the fine filtration cross-flow disc filter 10 enters the liquid storage box 14. The concentrate can be backflowed to the bottom of the container 6 through the backflow port 9 and mixed with the coarse filtration permeate. The filter screen aperture of the fine filtration cross-flow disc filter 10 is 10 μm.

[0096] The target content collection module 2 comprises a microporous membrane cross-flow disc filter 12 with a microporous membrane filter screen. The permeate filtered through the fine filtration module 4 can be conveyed to the microporous membrane cross-flow disc filter 12 by a second conveying pump 13. The target content is collected in the concentrate of the microporous membrane cross-flow disc filter 12 after being filtered through the microporous membrane filter screen.

[0097] The filter screen aperture of the microporous membrane cross-flow disc filter 12 is 0.1 μm. The microporous membrane is a hydrophilic polyether sulfone membrane with an opening rate of 85%. The effective filtration area of the cross-flow disc is 155 cm 2 , and the ratio of the effective filtration area to the overall area of the cross-flow disc is 0.5. The cross-sectional area of the inlet pipe is about 35 mm 2 , and the cross-sectional area of the flow channel of the cross-flow disc is about 15 mm 2 .

[0098] The permeate liquid filtered by the micro-porous membrane cross-flow disc filter 12 enters the waste liquid box 15, and the concentrated liquid is returned to the storage box 14 of the fine filtration module through the pipeline. During operation, the target content is collected in the concentrated liquid of the micro-porous membrane cross-flow disc filter 12 after circulating filtration for 5 minutes. The sample can be completely filtered and collected in about 60 minutes. After freeze-drying treatment, 15 grams of dry powder is obtained, and the bacterial content of the dry powder is 10 11 / 1g of dry powder.

[0099] The collected concentrated liquid of the micro-porous membrane cross-flow disc filter 12 is observed under a 200-fold microscope, as shown in Figure 3 .

[0100] The micro-porous membrane cross-flow disc filter 12 adopts a spiral structure, as shown in Figure 7 . The spiral structure design is relatively compact, which can be beneficial to reduce the volume of the overall equipment, and also has a good filtering effect.

[0101] An outer shell 16 can be arranged outside the container 6 of the stirring coarse filtration module 3, and an exhaust port 17 can be arranged at the top of the outer shell 16. The odor gas generated by the feces is discharged through the exhaust port 17 and removed by the deodorizing device 18.

[0102] Comparative Example 1

[0103] A device for separating and recovering intestinal contents, compared with the device for separating and recovering intestinal contents in Example 1, only has a impurity removal module, as shown in Figure 2 . The impurity removal module includes a stirring coarse filtration module 201 and a fine filtration module 202. The stirring coarse filtration module 201 is the same as the stirring coarse filtration module 3 in Example 1, which is a container 204 with a stirring device 203 at the upper part, and a coarse filtration screen 205 is arranged at the lower part of the container 204. After the feces and water are mixed in the container 204, the feces are fully dispersed by the stirring device 203, and the larger impurities in the feces are removed through the coarse filtration screen 205. The fine filtration module 202 includes a fine filtration screen 206, which uses a conventional filtering mode (non-cross-flow filtration) to remove smaller impurities in the feces. Since the pore size of the fine filtration screen is relatively small, the permeation speed of the filtrate is very slow during the filtration process, and a small amount of sample will block the fine filtration screen, which needs to be frequently replaced or cleaned, otherwise the filtrate is difficult to permeate.

[0104] The feces suspension after impurity removal by the fine filtration screen 206 is separated by centrifugation, 1500 rpm, 3 minutes, repeated three times, and the sample picture after each centrifugation is shown in Figure 8 . The supernatant after each centrifugation is analyzed by electron microscopy, as shown in Figure 9 . The Figure 8 and Figure 9As can be seen, when the obtained sample is centrifuged, the supernatant contains a large number of bacteria. The loss of these bacteria leads to a reduction in the effective amount of the final extract of the intestinal contents, and the distribution of bacteria is different from that in the original intestinal contents.

[0105] The final lower sediment after three centrifugations totaled approximately 6g. 2g of the solid precipitate was dissolved in 25ml of physiological saline. The image was taken under a 200x microscope. Figure 4 As shown. From Figure 4 It can be seen that the microbial content in the collected lower sediment is very low. Therefore, centrifugation is ineffective in collecting microorganisms, and the low microbial content in the product results in poor therapeutic effects.

[0106] As can be seen, compared with Comparative Example 1, Example 1 uses cross-flow filtration for fine filtration, removing smaller impurities from the fecal sample. The tangential force parallel to the membrane surface generated by the water flow continuously washes away trapped material from the membrane surface, preventing pore blockage, improving separation efficiency, and extending filter life. After the fine filtration step, microporous membrane cross-flow filtration is used to enrich the target intestinal contents, replacing traditional centrifugal separation technology. This allows for more precise and controllable extraction, high separation efficiency, and is also simple to operate and highly automated.

[0107] Comparative Example 2

[0108] An intestinal contents separation and recovery device includes: an impurity removal module and a target contents collection module. The impurity removal module includes a stirring coarse filtration module and a fine filtration module. The stirring coarse filtration module is a container with a stirring device at the top and a coarse filter screen at the bottom. The bottom of the container is provided with an outlet and a reflux outlet. After 130 grams of feces and 750 ml of physiological saline are mixed in the container, they are fully dispersed by the stirring device and filtered through the coarse filter screen to remove larger impurities from the feces.

[0109] The fine filtration module includes two layers of fine filter screens with different pore sizes. The pore sizes of the fine filter screens are 0.5 mm and 10 μm from top to bottom, respectively. The liquid after coarse filtration is transported to the fine filter screen, where finer impurities are removed. The fine filtration module also includes a liquid storage box, into which the permeate after filtration by the fine filter screen enters.

[0110] The target content collection module is the same as that in Example 1. However, due to the traditional multi-stage filtration method used in the fine filtration module, the filtration efficiency is very low. It takes approximately 150 minutes to filter and collect all samples, a significant increase compared to Example 1, yielding approximately 7g of the target product. The concentrated solution collected from the microporous membrane cross-flow disc filter was observed under a 200x microscope. Figure 5The collected microorganism content is increased compared with Comparative Example 1, but is still much lower than Example 1. In addition, it can be noted that the impurity content in Comparative Example 2 is significantly higher than that in Example 1.

[0111] It can be seen that, compared with Comparative Example 2, the fine filtration module and the target collection module in Example 1 both use cross-flow filtration, which uses the tangential force parallel to the membrane surface generated by the water flow to continuously flush the retained substances on the membrane surface, thereby helping to prevent the small pores from being blocked. In addition, since the retained substances on the surface are continuously flushed, it also helps to separate the microorganisms from the retained impurities, thereby improving the filtration efficiency and greatly increasing the probability of the microorganisms passing through the filter screen. The whole recovery and extraction process of Comparative Example 2 takes a long time, the filter screen is easily blocked during the filtration process, and the amount of the target product obtained is also small.

[0112] Example 2-5

[0113] According to another embodiment 2 of the present application, an intestinal content separation and recovery device is provided, which has the same overall structure as that of Example 1. The pore size of the filter screen of the fine filtration cross-flow disc filter 10 is 5 μm, and the pore size of the filter screen of the micro-porous membrane cross-flow disc filter 12 is 1 μm. The comparison chart of the particle size distribution of the concentrated liquid separated by Example 1 and Example 2 is shown in Figure 6 It can be seen that, by designing the pore sizes of the filter screens of the fine filtration module and the target content collection module, the target content in the concentrated liquid in Example 2 is enriched in different sizes, and the particle size distribution of the target content in the concentrated liquid in Example 2 is narrower.

[0114] According to another embodiment 3 of the present application, an intestinal content separation and recovery device is provided, which has the same overall structure as that of Example 1. The pore size of the filter screen of the fine filtration cross-flow disc filter 10 is 100 μm, and the pore size of the filter screen of the micro-porous membrane cross-flow disc filter 12 is 0.05 μm.

[0115] According to another embodiment 4 of the present application, an intestinal content separation and recovery device is provided, which has the same overall structure as that of Example 1. The pore size of the filter screen of the fine filtration cross-flow disc filter 10 is 50 μm, and the pore size of the filter screen of the micro-porous membrane cross-flow disc filter 12 is 20 μm.

[0116] According to another embodiment 5 of the present application, an intestinal content separation and recovery device is provided, which has the same overall structure as that of Example 1. The pore size of the filter screen of the fine filtration cross-flow disc filter 10 is 5 μm, and the pore size of the filter screen of the micro-porous membrane cross-flow disc filter 12 is 0.05 μm.

[0117] Example 6 and Comparative Example 3

[0118] The same intestinal content (feces) is equally divided into three parts by weight, marked as DY, MY and F, DY is used for Example 6, MY is used for Example 3, and F is the original feces.

[0119] Example 6 Separation and recovery of intestinal contents, using the same separation and recovery device as in Example 1, comprising:

[0120] Step 1) Dispersion: mixing the intestinal contents with a liquid dispersion medium, the ratio of intestinal contents to liquid dispersion medium (normal saline) being 100 g:500 ml, and dispersing the resulting mixture by mechanical means, with sufficient agitation by the stirring device 5, to form a dispersion mixture;

[0121] Step 2) Impurity removal: filtering the dispersion mixture through a filter screen structure, said filtering comprising coarse filtering and cross-flow fine filtering, the dispersion mixture in the container 6 being filtered through the coarse filter screen 7, and the filtrate being discharged through the outlet 8, to remove larger particulate impurities from the feces;

[0122] The coarse filter permeate enters the fine filter cross-flow disc filter 10, with a filter screen pore size of 50 μm, to remove finer impurities;

[0123] In which, after 5 min of agitation by the stirring device 5, sufficient permeate is obtained to begin the fine filtering operation; after 10 min of fine filtering, sufficient permeate (filtrate) is obtained to begin the collection step of the target contents;

[0124] Step 3) Collection: filtering the permeate of Step 2) through a micro-porous membrane cross-flow filter, to obtain a concentrate and a filtration waste liquid, said concentrate being used to collect the target contents;

[0125] The micro-porous membrane of the micro-porous membrane cross-flow filter has a non-uniform pore structure, with a pore size range of 0.2 μm, and an open porosity of 80%, said micro-porous membrane being a hydrophilic membrane; the effective filter area of the cross-flow disc is 155 cm 2 ; the cross-sectional area of the inlet pipe is about 35 mm 2 , and the cross-sectional area of the cross-flow disc flow channel is about 15 mm 2 ;

[0126] Step 4) Freeze-drying: freeze-drying the target contents obtained in Step 3) to form a dry powder 12 g, with a bacterial content of 1.5 x 10 11 per 1 g of dry powder. Capsules are prepared.

[0127] In which, the concentrate of the fine filtering of the impurity removal step is returned to mix with the coarse filter permeate, to form a circulation loop; the concentrate of the micro-porous membrane cross-flow filter is returned to mix with the permeate of the fine filtering of the impurity removal step, to form a circulation loop.

[0128] The process of Step 1) to Step 3), i.e. the entire separation and recovery process, takes 60 min.

[0129] Comparative Example 3:

[0130] The intestinal content separation and recovery device as shown in Figure 2 The intestinal content separation and recovery device as shown in

[0131] Example 6 (DY) and Comparative Example 3 (MY) were repeated 4 times (4 times of experiments were performed on the same donor's feces within a week), and the obtained target contents and original feces (F) were subjected to 16sR DNA sequencing to determine the species and abundance of the bacteria.

[0132] In community ecology, the diversity of microorganisms is studied by analyzing the diversity of a single sample (Alpha diversity), which can reflect the abundance and diversity of microbial communities, including a series of statistical analysis indices to estimate the species abundance and diversity of the environment community. Shannon: one of the indices for estimating the diversity of microorganisms in a sample. The greater the Shannon value, the higher the community diversity.

[0133] Figure 9 It is shown that the species diversity of the extracted flora of the present application is better than that of the centrifugal method, and there is a significant difference between the two methods.

[0134] PCA analysis (Principal Component Analysis), i.e., principal component analysis, uses variance decomposition to reflect the differences in multiple sets of data on a two-dimensional coordinate graph, and the coordinate axes take two characteristic values that can best reflect the variance. For example, the more similar the sample composition, the closer the distance reflected in the PCA graph.

[0135] Figure 10 - Figure 12 It is shown that the extracted flora of the present application is closer to the original feces in terms of species and abundance, and the degree of similarity is higher, and the statistical difference is significant.

[0136] The target contents obtained from Example 6 (DY) and Comparative Example 3 (MY) and the original feces (F) have a significant statistical difference.

[0137] R-value is between (-1, 1), and R-value is greater than 0, indicating that the difference between the groups is significant. The reliability of statistical analysis is represented by P-value, and P<0.05 indicates that the statistics are statistically significant.

[0138] Table 1 Statistical values of the target contents obtained from Example 6 (DY) and Comparative Example 3 (MY) and the original feces (F)

[0139] Table 1 Statistical values of the target contents obtained from Example 6 (DY) and Comparative Example 3 (MY) and the original feces (F) Group R-value P-value DY-MY 1 0.036 F-MY 1 0.035 F-DY 0.9896 0.033

[0140] It can be seen that the target content (flora) obtained by the method of the present application is closer to the species and distribution of the flora in the initial intestinal content (feces) compared with the centrifugal method. At the same time, the method of the present application has better repeatability.

[0141] Example 7 and Comparative Examples 4-5

[0142] The well-mixed intestinal content (feces) was evenly divided by weight into 9 parts, each weighing 100 g, 3 parts for Example 7, 3 parts for Comparative Example 4, and 3 parts for Comparative Example 5.

[0143] Example 7 intestinal content separation and recovery, using the same separation and recovery device and method as Example 1, the only difference is that:

[0144] The physiological saline added to the container 6 was 500 ml; the coarse filter screen aperture was 2 mm; the fine filter module 4 filter screen aperture was 40 μm; the microporous membrane cross-flow disc filter 12 filter screen aperture was 0.5 μm, and the opening rate was 85%.

[0145] The separation and recovery was completed in 60 minutes, and the obtained concentrated bacterial solution was freeze-dried to obtain bacterial powder, 3 times, 12 g, 11 g, and 12 g, respectively.

[0146] Comparative Example 4, referring to Example 3 of Chinese Patent Application CN201810852561.2, the only difference is that the physiological saline is 500 ml, and the fine filter screen aperture from top to bottom is 0.5 mm and 40 μm, respectively.

[0147] The separation and recovery was completed in 120 minutes, and the obtained concentrated bacterial solution was freeze-dried to obtain bacterial powder, 3 times, 8 g, 8 g, and 7 g, respectively.

[0148] Comparative Example 5, referring to the manner of Comparative Example 1, the only difference is that the physiological saline added to the container 6 is 550 ml; the coarse filter screen aperture is 2 mm; and the fine filter module 4 filter screen aperture is 40 μm.

[0149] The separation and recovery was completed in 150 minutes, and the obtained concentrated bacterial solution was freeze-dried to obtain bacterial powder, 3 times, 3 g, 2 g, and 2 g, respectively.

[0150] The average copy number of the obtained samples of Example 7 and Comparative Examples 4, 5 was determined, which reflects the number of biological samples in the sample.

[0151] The results of the average copy number / g of dry powder of the samples of Example 7 and Comparative Examples 4, 5 are shown in Table 2.

[0152] Table 2 Average copy number per g of dry powder

[0153]

[0154] The values of average copy number per g of dry powder in Table 2 were logarithmized, as shown in Table 3. Figure 13

[0155] From the results of Example 7, Comparative Example 4 and Comparative Example 5, it can be concluded that the results obtained by the method of the present application are of the same order of magnitude, have good repeatability, and have high efficiency of separation and recovery, and can be completed in 1 h. The quality of the bacterial flora dry powder obtained is higher. Although Comparative Example 4 of the prior art has the same order of magnitude and good repeatability, the filtration process still uses conventional filtration, and the nuclear pore membrane is used for cross-flow collection. The time for the entire separation and recovery is long, and the quality of the bacterial flora dry powder obtained is not as high as that of the present application, and the average copy number per g of dry powder is 3 orders of magnitude lower than that of the present application. Comparative Example 5 uses multi-stage conventional filtration, and the results obtained are significantly different, the repeatability is poor, the time for the entire separation and recovery is longer, the quality of the bacterial flora dry powder obtained is less, and the average copy number per g of dry powder is more than 8 orders of magnitude lower than that of the present application.

[0156] The above detailed the embodiments of the present application, but only for the purpose of facilitating understanding and illustration, and should not be regarded as limiting the present application. Although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art will understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent replacements can be made to part of the technical features; and these modifications or replacements are also regarded as falling within the spirit and scope of the technical solutions described in the embodiments of the present application.​

Claims

1. A device for separating and recovering intestinal contents, characterized in that, The separation and recovery device includes a purification module and a target content collection module. The purification module removes impurities from the intestinal contents and includes a coarse filtration module and a fine filtration module. The coarse filtration module has an outlet and a return outlet. The fine filtration module includes a fine cross-flow disc filter and a storage box. The coarse permeate filtered by the coarse filtration module is filtered by the fine cross-flow disc filter to obtain finely filtered permeate and concentrate. The finely filtered permeate is sent to the storage box, and the finely filtered concentrate is returned to the coarse filtration module through the return outlet via a pipeline to mix with the coarse permeate, forming a circulation loop. The target content collection module mainly... The system comprises a microporous membrane cross-flow filter. The microporous membrane has a sponge-like porous structure with a pore size ranging from 0.01 to 20 μm and an open porosity of over 30%. The microporous membrane is a hydrophilic membrane. The microporous membrane cross-flow filter is a microporous membrane cross-flow disc filter with a microporous membrane filter screen. The cross-sectional area of ​​the flow channel of the microporous membrane cross-flow filter is smaller than the cross-sectional area of ​​the inlet pipe. The target contents collection module is connected to the storage box via a pipe. The permeate from the fine filtration is sent to the target contents collection module to obtain a concentrated solution. The concentrated solution in the target contents collection module flows back to the storage box via a pipe and mixes with the permeate in the fine filtration module to form another circulation loop.

2. The apparatus according to claim 1, characterized in that, The microporous membrane cross-flow filter has a spiral fluid channel inside; the ratio of the effective filtration area of ​​the cross-flow plate to the total area of ​​the cross-flow plate is 0.3-0.

9.

3. The apparatus according to claim 1 or 2, characterized in that, The flow channel of the microporous membrane cross-flow filter is a spiral cross-flow channel.

4. The apparatus according to claim 1 or 2, characterized in that, Microporous membranes include one or more of polyethersulfone membranes, cellulose membranes, and polymethacrylate membranes, which may be modified or unmodified.

5. The apparatus according to claim 1 or 2, characterized in that, The pore size of the microporous membrane ranges from 0.1 to 10 μm, and the porosity of the microporous membrane is above 50%.

6. A method for separating and recovering intestinal contents, characterized in that, include: Step 1) Dispersion: Mix the intestinal contents with a liquid dispersion medium at a ratio of 100g: 200-5000ml. Disperse the resulting mixture mechanically to form a uniformly dispersed mixture. Step 2) Impurity Removal: The dispersed mixture is filtered through a filter structure to remove impurities. The filtration includes coarse filtration and fine filtration. Coarse filtration yields coarse permeate, followed by fine filtration, which is cross-flow filtration, to obtain permeate and concentrate. The concentrate from the fine filtration is refluxed and mixed with the coarse permeate to form a circulation loop. The mesh structure for fine filtration is a fine cross-flow disc filter. Step 3) Collection: The permeate from step 2) is filtered through a microporous membrane cross-flow filter to obtain a concentrate and a filtration waste liquid. The concentrate is used to collect the target contents. The concentrate from step 3) is refluxed and mixed with the permeate from step 2) to form another circulation loop. The microporous membrane cross-flow filter has a sponge-like porous structure, with a pore size ranging from 0.01 to 20 μm and an open porosity of over 30%. The microporous membrane is a hydrophilic membrane, and the microporous membrane cross-flow filter is a microporous membrane cross-flow disc filter of microporous membrane filter screen. The cross-sectional area of ​​the flow channel of the microporous membrane cross-flow filter is smaller than the cross-sectional area of ​​the inlet pipe.

7. The method according to claim 6, characterized in that, The concentration of microorganisms in the dispersion mixture of step 1) is at least 10. 8 per ml.

8. The method according to claim 6 or 7, characterized in that, The ratio of the intestinal contents to the liquid dispersion medium is 100g: 300-1000ml, the pore size of the microporous membrane is in the range of 0.1-10μm, and the porosity of the microporous membrane is above 50%.

9. A composition comprising an extract of intestinal contents, characterized in that, The intestinal contents extract is prepared using the intestinal contents separation and recovery device according to any one of claims 1-5 or by the intestinal contents separation and recovery method according to any one of claims 6-8, and the size of the obtained intestinal contents extract is 0.01-50 μm.

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