In-situ capture and enrichment device for actinomycetes in lakes and reservoirs

Abstract

The invention discloses an in-situ capture and enrichment device for lake and reservoir actinomycetes, and belongs to the technical field of water treatment. The lake and reservoir actinomycete in-situ capture and enrichment device comprises a separation and enrichment assembly used for conducting in-situ enrichment on actinomycetes, the separation and enrichment assembly comprises a fiber membrane container, a hollow fiber membrane rubber cover and filling and sealing pipes, a hollow fiber membrane is arranged in the fiber membrane container, the filling and sealing pipes are arranged at the two ends of the fiber membrane container, the ends are sealed through the rubber cover, and the hollow fiber membrane is arranged in the fiber membrane container. The device further comprises a fixing device used for fixing the separation and enrichment assembly, the fibrous membrane container is vertically clamped to the fixing device, the separation and enrichment assembly is combined with the culture medium, growth of actinomycetes is met while growth of other competitive infectious microbes in an inhibition area is achieved, and therefore the in-situ capture and enrichment rate of the actinomycetes is increased.

Description

Technical Field

[0001] The present invention relates to the technical field of water treatment, and particularly relates to a device for in-situ capture and enrichment of actinomycetes in lakes and reservoirs. Background Art

[0002] At present, the problem of off-flavors in water source reservoirs has become one of the most important problems of endogenous pollution in reservoirs. Whether drinking water has an off-flavor is the most direct criterion for consumers to measure the safety of drinking water. The water quality indicators stipulated in the "Sanitary Standard for Drinking Water in China" have been adjusted from the previous 106 items to 97 items, and 2-methylisoborneol and geosmin are two newly added off-flavor indicators. Odor and taste are important sensory property evaluation indicators, and the standard stipulates that: the sensory properties of drinking water are good, without abnormal odor or taste. Compounds causing off-flavors in water sources can be detected at trace levels, and even if the content is only at the nanogram level, it will have an impact on the human body.

[0003] In lake and reservoir waters, actinomycetes are the main producers of 2-methylisoborneol and geosmin, and their odor production is higher than that of cyanobacteria during non-algal bloom periods. In addition, actinomycetes play a crucial role in the ecosystem and are widely regarded as an effective source of natural compounds related to drugs. Actinomycetes have high diversity, but about 95% of them are still unknown to science. The reporting rate of actinomycetes is very low, mainly because most actinomycetes cannot grow in artificial culture media, and the laboratory cannot provide an environment similar enough to their natural habitats. In addition, many microbial species in nature are in a state of low metabolic activity and cannot easily proliferate without appropriate resuscitation. The previously developed culturomics techniques were mainly designed and applicable for bacteria, and due to the cell size and hyphal growth characteristics of actinomycetes, few attempts have been made to isolate actinomycetes.

[0004] In the existing microbial enrichment devices, the plate enrichment culture method requires collecting a large amount of water samples and transporting them to the laboratory, and the subsequent laboratory treatment steps are cumbersome. Water sample contamination and actinomycete succession are likely to occur during the transportation process and laboratory treatment process, affecting the purity of actinomycetes. Summary of the Invention

[0005] The object of the present invention is to overcome the problems in the prior art and provide a device for in-situ capture and enrichment of actinomycetes in lakes and reservoirs, so as to better capture and enrich actinomycetes in-situ.

[0006] The present invention provides an in-situ capture and enrichment device for lake reservoir actinomycetes, and a separation and enrichment component for in-situ enrichment of actinomycetes. The separation and enrichment component includes a fiber membrane container, a hollow fiber membrane, a rubber cap, and a potting tube. The hollow fiber membrane is placed in the fiber membrane container. The potting tubes are provided at both ends of the fiber membrane container, and the ends are sealed by the rubber cap. There are gaps on the outer wall of the fiber membrane container to allow water to enter the interior of the fiber membrane container to contact the hollow fiber membrane. The device also includes a fixing device for fixing the separation and enrichment component, and the fiber membrane container is vertically clamped on the fixing device.

[0007] Preferably, the fiber membrane container is made of optically transparent pure polystyrene.

[0008] Preferably, a plurality of the hollow fiber membranes are provided and evenly arranged in the fiber membrane container.

[0009] Preferably, the fixing device includes a holding tray, a support column, and a top cover. The holding tray and the top cover are both set as disc structures. The support column is vertically inserted between the holding tray and the top cover. A first card slot is opened in the center of the middle part of the holding tray, and one end of the support column is clamped in the first card slot and the other end is fixedly connected to the top cover.

[0010] Preferably, a plurality of the fiber membrane containers are provided, and the plurality of fiber membrane containers are evenly arranged around the axis of the support column. A plurality of second card slots corresponding to the plurality of fiber membrane containers one by one are opened on the holding tray, and the second card slots are used for installing the fiber membrane containers.

[0011] Preferably, the holding tray, the support column, and the top cover are all made of transparent pure polystyrene.

[0012] Preferably, a plurality of hook rings are fixedly connected to the top of the top cover, and a traction rope is sleeved on the plurality of hook rings, and the plurality of traction ropes converge directly above the center of the circle of the top cover.

[0013] Compared with the prior art, the beneficial effects of the present invention are as follows: The device of the present invention lowers the fixed component into the lake water according to the reservoir and enrichment requirements. The fixed component supports and stores the fiber membrane container, which is conducive to observing the enrichment and growth status of actinomycetes, realizing in-situ enrichment of lake reservoir actinomycetes, avoiding the pollution and reduction of microbial activity caused by the transportation process in the traditional method. The hollow fiber membrane can intercept pollutants and cover them on the culture medium, providing attachment sites for the growth of actinomycetes and not hindering the extension of actinomycete hyphae to the culture medium, improving the in-situ capture and enrichment efficiency of actinomycetes. At the same time, in order to fully adapt to the nutritional characteristics of lake reservoir actinomycetes, while meeting the growth of actinomycetes, it can also inhibit the growth of other competing miscellaneous bacteria in the area. The culture medium is injected into the hollow fiber membrane. The present device adopts the method of in-situ enrichment, maintaining the physical contact and metabolite exchange between microorganisms, improving the efficiency of enriching actinomycete flora that depends on symbiosis, while enhancing the culturable ability of unculturable actinomycetes, and at the same time avoiding the pollution phenomenon caused by the transportation process and laboratory operations. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 It is a schematic diagram of the overall structure of the present invention.

[0015] Figure 2 It is a schematic diagram of the structure of the fiber membrane container of the present invention.

[0016] Figure 3 It is a schematic diagram of the structure of the hollow fiber membrane of the present invention.

[0017] Figure 4 It is a schematic diagram of the structure of the rubber cap of the present invention.

[0018] Figure 5 It is a schematic diagram of the structure of the holding tray of the present invention.

[0019] Figure 6 It is an enrichment culture effect diagram of Actinomyces on Gao's No. 1 and Modified Gao's No. 1 of the present invention.

[0020] Figure 7 It is a comparison diagram of the culture effects of Actinomyces on Gao's No. 1 and Modified Gao's No. 1 of the present invention.

[0021] Description of reference numerals: 1. towing rope; 2. top cover body; 3. support column; 4. fiber membrane container; 5. holding tray; 6. hook ring; 7. first card slot; 8. second card slot; 9. hollow fiber membrane; 10. potting tube; 11. rubber cap. DETAILED DESCRIPTION OF THE INVENTION

[0022] The following combines the attached Figures 1 to 7, for the purpose of making the objectives, technical solutions and advantages of the embodiments of the present invention clearer, the following will, with reference to the accompanying drawings of the embodiments of the present invention, clearly and completely describe the technical solutions of the embodiments of the present invention. Obviously, the described embodiments are a part rather than all of the embodiments of the present invention. Based on the described embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the scope of protection of the present invention. Unless otherwise defined, the technical terms or scientific terms used herein shall have the ordinary meanings understood by those with ordinary skills in the field to which the present invention pertains.

[0023] The terms "first", "second" and similar terms used in the description of the specification and claims of this invention for patent application do not denote any order, quantity or importance, but are only used to distinguish different components. Terms such as "comprising" or "including" mean that the elements or items appearing before this word cover the elements or items listed after this word and their equivalents, without excluding other elements or items. Terms such as "inside", "outside", "above", "below", "far", "near", "front", "rear", etc. are only used to indicate relative position relationships. When the absolute position of the object being described changes, the relative position relationship may also change accordingly. The drawings in the present invention are not strictly drawn to actual scale, and the specific dimensions and quantities of each structure can be determined according to actual needs. The drawings described in the present invention are only schematic diagrams of the structures.

[0024] An in-situ capture and enrichment device for lake reservoir actinomycetes, as Figures 1 to 5 shown, includes a separation and enrichment component for in-situ enrichment of actinomycetes. The separation and enrichment component includes a fiber membrane container 4, a hollow fiber membrane 9, a rubber cap 11 and a potting tube 10. The hollow fiber membrane 9 is placed in the fiber membrane container 4. The fiber membrane container 4 is provided with the potting tubes 10 at both ends, and the ends are sealed by the rubber cap 11. There are gaps on the outer wall of the fiber membrane container 4 to allow water to enter the inside of the fiber membrane container 4 to contact the hollow fiber membrane 9. It further includes a fixing device for fixing the separation and enrichment component. The fiber membrane container 4 is vertically clamped on the fixing device.

[0025] The device of the present invention lowers the fixed component into the lake water according to the reservoir and enrichment requirements. The fixed component supports and stores the fiber membrane container 4, which is beneficial to observing the enrichment and growth status of actinomycetes, realizing the in-situ enrichment of lake reservoir actinomycetes, avoiding the pollution and the reduction of microbial activity caused by the transportation process in the traditional method. The hollow fiber membrane 9 is provided with filter holes, which can intercept pollutants and cover them on the culture medium, provide attachment sites for the growth of actinomycetes, and do not prevent the actinomycetes hyphae from extending towards the culture medium, improving the in-situ capture and enrichment efficiency of actinomycetes. At the same time, in order to fully adapt to the nutritional characteristics of lake reservoir actinomycetes, while meeting the growth of actinomycetes, it can also inhibit the growth of other competitive miscellaneous bacteria in the area. The culture medium is injected into the hollow fiber membrane. The inner diameter of the hollow fiber membrane filament is 1.2 mm, the outer diameter is 2.0 mm, and the filtration pore diameter is 0.1 μm. While intercepting pollutants and covering them on the culture medium and providing attachment sites for the growth of actinomycetes, it does not prevent the actinomycetes hyphae from extending towards the culture medium.

[0026] The present invention utilizes the growth characteristics and carbon source preference of actinomycetes, adopts the improved Gao's No. 1 medium to provide an environment for the growth and enrichment of actinomycetes, uses the hollow fiber membrane 9 as the support system for the growth of actinomycetes, and induces a large amount of in-situ enrichment of actinomycetes in the lake reservoir water on the hollow fiber membrane 9. The in-situ separation and enrichment device for lake reservoir actinomycetes provided by the present invention is applied to the resource utilization of lake reservoir microorganisms, can efficiently enrich known and unknown actinomycete flora, and solve the problem that a large number of actinomycetes in lake reservoirs are difficult to be in-situ enriched and are not conducive to laboratory isolation and culture at a lower economic cost. This device adopts the method of in-situ enrichment, while maintaining the physical contact and metabolite exchange between microorganisms, improving the efficiency of enriching actinomycete flora that depends on symbiosis, enhancing the culturing ability of difficult-to-culture actinomycetes, and at the same time avoiding the pollution phenomenon caused by the transportation process and laboratory operations.

[0027] Preferably, as Figures 1 to 3 shown, the fiber membrane container 4 is made of transparent pure polystyrene, and a number of hollow fiber membranes 9 are provided and arranged in the fiber membrane container 4.

[0028] The fiber membrane container 4 is made of transparent pure polystyrene, which has strong economic applicability and low cost. The inner diameter of the hollow fiber membrane 9 is 1.2 mm and the length is 20 cm. In the present invention, the culture medium is the improved Gao's No. 1 medium, and the liquid-state improved Gao's No. 1 medium can be added into it with a syringe. After cooling to a solid state, it is reserved. The solid-state improved Gao's No. 1 medium and the hollow fiber membrane 9 are neatly arranged in the fiber membrane container 4 with a height of 20 cm and a diameter of 4 cm, and are sealed with a potting tube 1010 and a rubber cap 1111.

[0029] Preferably, as Figures 1 to 5As shown in the figure, the fixing device includes a holding tray 5, a support column 3, and a top cover 2. Both the holding tray 5 and the top cover 2 are set as disc structures. The support column 3 is vertically inserted between the holding tray 5 and the top cover 2. A first card slot 7 is opened at the center of the middle part of the holding tray 5. One end of the support column 3 is clamped in the first card slot 7. A number of fiber membrane containers 4 are provided. The number of fiber membrane containers 4 is arranged in an array around the axis of the support column 3. A number of second card slots 8 corresponding to the number of fiber membrane containers 4 are opened on the holding tray 5. The second card slots 8 are used for installing the fiber membrane containers 4.

[0030] The holding tray 5 can be connected to the support column 3 through the first one located at the center. The support column 3 is slightly longer than the fiber membrane container 4 to ensure that the top end of the fiber membrane container 4 just contacts the top cover 2, so as to achieve the effect of fixing the fiber membrane container 4. The fiber membrane container 4 can be inserted into the second card slot 8 opened on the holding tray 5 to achieve the purpose of fixation. The diameter of the second card slot 8 corresponds to the diameter of the fiber membrane container 4, which is convenient for the disassembly of the separation and enrichment component and is beneficial to observing the enrichment and growth status of actinomycetes.

[0031] Preferably, as Figures 1 to 5 shown, the holding tray 5, the support column 3, and the top cover 2 are all made of transparent pure polystyrene.

[0032] In the present invention, the holding tray 5, the support column 3, and the top cover 2 are all made of transparent pure polystyrene. On the one hand, transparent pure polystyrene has strong economic applicability and low cost. On the other hand, because the transparent pure polystyrene material is light, the overall weight of the device can be reduced, thereby improving the overall stability of the device and better enriching actinomycetes in situ.

[0033] Preferably, as Figure 1 shown, a number of hook rings 6 are fixedly connected to the top of the top cover 2. A number of traction ropes 1 are sleeved on the number of hook rings 6. The number of traction ropes 1 converges directly above the center of the top cover 2.

[0034] Four evenly distributed hook rings 6 are arranged on the top of the top cover 2. The four hook rings 6 are respectively connected to the traction rope 1 and converge directly above the center of the disc of the fixing device, which is used for arranging and lifting the actinomycetes separation and enrichment component.

[0035] The culture medium in this example is the modified Gao's No. 1 culture medium, which is the modified Gao's No. 1 culture medium in the patent with the patent number CN202410778199.4 applied by the applicant. Its components include 1 part of soluble starch, 1 / 20 part of KNO3, 3 / 50 part of aspartic acid, 1 / 50 part of methyl pyruvate, 1 / 20000 part of FeSO4·7H2O, 1 / 400 part of MgSO4·7H2O, 1 / 400 part of K2HPO4·3H2O, 1 / 400 part of NaCl, and 1 part of agar. Dissolve it in 50 parts of pure water in a water bath at 90 - 95 °C, and adjust the pH to 7.0 - 7.6. Sterilize it at 121 °C for 30 minutes and then reserve it for use.

[0036] Collect water samples from the surface layer (0 - 0.5 m), middle layer (20 - 25 m), and bottom layer (50 - 55 m) of the Li Jiahe Reservoir in Xi'an. Take 50 mL of the water sample and filter it through a 0.22 - μm polyester fiber membrane. Use sterile forceps to stick the filtered membrane onto the modified Gao's No. 1 solid culture medium and the Gao's No. 1 solid culture medium. Incubate the culture medium with the attached membrane in an incubator at 30 °C. After incubating upside - down for 7 days, the effect of plate enrichment culture is as Figure 6 shown. (a) - (c) are the average colony counts of the Gao's No. 1 solid culture medium in the surface, middle, and bottom waters of Li Jiahe respectively; (d) - (f) are the average colony counts of the modified Gao's No. 1 solid culture medium in the surface, middle, and bottom waters of Li Jiahe respectively. The average colony counts of the Gao's No. 1 solid culture medium in the surface, middle, and bottom waters of Li Jiahe are 340 CFU / L, 607 CFU / L, and 3140 CFU / L respectively. The average colony counts of the modified Gao's No. 1 solid culture medium in the surface, middle, and bottom waters of Li Jiahe are 547 CFU / L, 973 CFU / L, and 4100 CFU / L respectively as Figure 7 shown. The modified Gao's No. 1 solid culture medium has a higher enrichment effect on actinomycetes compared to the ordinary Gao's No. 1 culture medium.

[0037] The usage method of the in - situ capture and enrichment device for lake - reservoir actinomycetes of the present invention is as follows: Add the liquid - state modified Gao's No. 1 culture medium into it through a syringe, and reserve it after cooling to a solid state. Then, neatly arrange the hollow fiber membrane 9 in the fiber membrane container 4, and seal it with the potting tube 10 and the rubber cap 11. There are gaps on the outer wall of the fiber membrane container 4 to allow water to enter the container interior and contact the hollow fiber membrane 9.

[0038] Insert several groups of fiber membrane containers 4 into the second card slots 8 opened on the containing tray 5 for fixation, tie it to the hook ring 6 through the towing rope 1, and lift the whole device into the lake water for in - situ capture and in - situ enrichment of actinomycetes.

[0039] Although embodiments of the present invention have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the present invention, and the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A device for in-situ capture and enrichment of lake actinomycetes, characterized in that: include: A separation and enrichment component for in-situ enrichment of actinomycetes, the separation and enrichment component comprising a fiber membrane container (4), a hollow fiber membrane (9), a rubber cover (11) and a potting tube (10), wherein the hollow fiber membrane (9) is placed in the fiber membrane container (4), the potting tube (10) is provided at both ends of the fiber membrane container (4), and the ends are sealed by the rubber cover (11), and there is a gap in the outer wall of the fiber membrane container (4) to allow water to enter the interior of the fiber membrane container (4) so ​​as to contact the hollow fiber membrane (9); A fixing device is used to fix the separation and enrichment component, and the fiber membrane container (4) is vertically clamped on the fixing device.

2. The in-situ capture and enrichment device for lake and reservoir actinomycetes according to claim 1, characterized in that: The fiber membrane container (4) is made of transparent pure polystyrene.

3. The in-situ capture and enrichment device for lake and reservoir actinomycetes according to claim 1, characterized in that: A plurality of hollow fiber membranes (9) are arranged evenly in the fiber membrane container (4).

4. The in-situ capture and enrichment device for lake and reservoir actinomycetes according to claim 1, characterized in that: The fixing device comprises a receiving tray (5), a supporting column (3) and a top cover (2); the receiving tray (5) and the top cover (2) are both arranged as disc structures; the supporting column (3) is vertically inserted between the receiving tray (5) and the top cover (2); a first clamping groove (7) is provided at the center of the receiving tray (5); one end of the supporting column (3) is clamped in the first clamping groove (7) and the other end is fixedly connected to the top cover (2).

5. The in-situ capture and enrichment device for lake and reservoir actinomycetes according to claim 4, characterized in that: A plurality of the fiber membrane containers (4) are arranged, and the plurality of the fiber membrane containers (4) are evenly arranged around the axis of the support column (3). The containing tray (5) is provided with a plurality of second slots (8) corresponding one-to-one to the plurality of the fiber membrane containers (4), and the second slots (8) are used to install the fiber membrane containers (4).

6. The in-situ capture and enrichment device for lake actinomycetes as claimed in claim 4, characterized in that: The containing tray (5), the supporting column (3) and the top cover (2) are all made of transparent pure polystyrene.

7. The in-situ capture and enrichment device for lake and reservoir actinomycetes according to claim 4, characterized in that: A plurality of hook rings (6) are fixedly connected to the top of the top cover body (2), and a plurality of traction ropes (1) are sleeved on the hook rings (6). The plurality of traction ropes (1) converge at a point directly above the center of the top cover body (2).

Images