CN114990018A - A kind of domestication method and equipment of Bacillus subtilis used for improving bottom of aquatic products - Google Patents

Abstract

The invention discloses a domestication method and equipment of Bacillus subtilis used for bottom improvement of aquatic products, and relates to the technical field of bottom improvement of aquatic products. In order to solve the problem that the current bottom modification products on the market are not targeted, the bottom modification effect is general, specific Bacillus strains cannot be put in according to different pond bottom environments, and the organic matter in the sediment cannot be effectively oxidized. A method and equipment for domestication of Bacillus subtilis for aquaculture bottom improvement. By preparing a culture medium, the Bacillus subtilis is activated in the culture medium, which provides a good environment for the mechanism of further domesticating Bacillus in aquaculture wastewater, so that the culture The base can achieve the effect of high-density growth of organic matter in aquaculture wastewater, and has a specific cultivation effect on Bacillus, and gradually domesticate to obtain Bacillus strains that are specifically adapted to the environment at the bottom of aquaculture ponds, which can be more effective. The organic matter in the sediment is oxidized, thereby improving the bottom environment of the pond, which is highly targeted.

Description

A kind of domestication method and equipment of Bacillus subtilis used for improving bottom of aquatic products

technical field

The invention relates to the technical field of aquatic product bottom improvement, in particular to a domestication method and equipment of Bacillus subtilis used for aquatic product bottom improvement.

Background technique

Due to the lack of oxygen at the bottom of the pond, the ammonia nitrogen, nitrite and hydrogen sulfide in the pond exceed the standard, resulting in low aquaculture production and low output value. However, there are still the following problems in the current aquatic product reform:

1. The bottom-modifying products currently on the market are not targeted, and the bottom-modifying effect is general. It is impossible to put specific Bacillus strains according to the environment at the bottom of different ponds, and cannot effectively oxidize the organic matter in the sediment;

2. Aquaculture wastewater contains a variety of bacteria or sludge, and some microorganisms are easily inhibited by toxic substances in wastewater. It is often necessary to grow the sludge or bacteria in aquaculture wastewater before adding bottom-modified products. acclimatization over time, however, microorganisms in aquaculture wastewater are difficult to utilize and degrade;

3. Quantitative addition cannot be carried out during the configuration of culture medium and simulated wastewater. The process is not precisely measured, the amount of additive cannot be guaranteed, and the error is large. In addition, the concentration of the addition is often concentrated in one place, resulting in uneven concentration, which greatly affects domestication effect.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide a method and equipment for domestication of Bacillus subtilis for aquaculture bottom improvement. A good environment enables the medium to achieve the effect of high-density growth of organic matter in aquaculture wastewater, and has a specific cultivation effect on Bacillus, and gradually domesticates to obtain Bacillus strains that are specifically adapted to the environment at the bottom of aquaculture ponds , non-toxic and harmless, friendly to the environment, can more effectively oxidize the organic matter in the bottom mud, thereby improving the bottom environment of the pond, and has strong pertinence to solve the problems raised in the above background technology.

To achieve the above object, the present invention provides the following technical solutions:

A kind of domestication method of Bacillus subtilis for improving bottom of aquatic products, comprising the following steps:

S1: activate Bacillus, prepare a culture medium, and activate Bacillus subtilis in the culture medium;

S2: domesticate Bacillus, prepare simulated aquaculture wastewater, and gradually increase the content of simulated aquaculture wastewater into the culture medium;

S3: Obtain Bacillus, and gradually domesticate to obtain Bacillus strains that are specifically adapted to the environment at the bottom of the aquaculture pond.

Further, for S1, the substratum is made up of the following raw material components and parts by weight: the consumption of distilled water is 1L, the consumption of glucose is 20g, the consumption of peptone is 15g, the consumption of sodium chloride is 5g, and the consumption of beef extract is 5g. The dosage is 0.5g, and the amount of agar is 20g.

Further, for S1, the pH value of the culture medium is 7, and the temperature is 32 degrees Celsius.

Further, for S2, the preparation of the simulated aquaculture wastewater includes the following steps:

S201: take f/2 medium as the base liquid, add ammonium chloride and sodium dihydrogen phosphate, so that the initial PO ₄ ^3- -P(NaH ₂ PO ₄ ) concentration of simulated aquaculture wastewater is 1.5 mg/L, NH ₄ +- N(NH ₄ Cl) concentrations were 0 mg/L (f/2 medium), 20 mg/L, 40 mg/L, 80 mg/L, 100 mg/L, and COD (glucose) concentrations were 500 mg/L.

Further, for S201, the simulated aquaculture wastewater is added with a trace element liquid in the preparation process, and the trace element liquid is composed of the following raw material components and parts by weight: the consumption of H ₃ BO ₃ is 2.9*10 ^-3 g/L , the dosage of MnCI ₂ -4H ₂ O is 1.8*10 ^-5 g/L, the dosage of ZnSO ₄ -7H ₂ O is 2.2*10 ^-4 g/L, and the dosage of Na ₂ MoO ₄ -2H ₂ O is 3.9* 10 ^-4 g/L, the dosage of CuSO ₄ -5H ₂ O is 8.0*10 ^-5 g/L, and the dosage of Co(NO ₃ ) ₂ -6H ₂ O is 5.0*10 ^-5 g/L.

The present invention provides another technical solution, a device for the domestication method of Bacillus subtilis for aquaculture bottom improvement, including an incubator for placing culture medium and preparing simulated aquaculture wastewater, the center of the upper surface of the incubator is connected to the center of the incubator through bolts. The hydraulic barrel is fixedly connected, the hydraulic barrel is set as a transparent glass cylinder, the side wall of the hydraulic barrel is provided with a scale, the lower surface of the inner wall of the incubator is fixedly connected to the culture dish through bolts, and the center of the upper surface of the inner wall of the incubator is connected to the upper liquid inlet pipe through bolts Fixed connection, the lower end of the upper liquid inlet pipe is nested with the lower liquid inlet pipe, the upper surface of the inner wall of the incubator is fixedly connected to the guide column through bolts, the lower surface of the guide column is fixedly connected to the surrounding of the support plate through bolts, and the guide column is set to telescopic The column, the center of the support plate is fixedly connected with the outer wall of the lower liquid inlet pipe, and the outer wall of the culture dish is sleeved with a heating wire.

Further, the lower surface of the lower liquid inlet pipe is rotatably connected to the shunt box through a bearing, the surrounding of the shunt box is fixedly connected to the shunt conduit through bolts, the lower surface of the inner wall of the shunt box is rotatably connected to the shunt shaft through a bearing, and the side walls of the shunt shaft are respectively provided with There are evenly distributed split fan blades, the lower surface of the lower liquid inlet pipe is respectively provided with liquid outlets, the liquid outlets are respectively inclined and the center of the lower surface of the lower liquid inlet pipe is symmetrically distributed and inclined in the same direction.

Further, the center of the lower surface of the shunt box is rotatably connected to the transmission shaft through a bearing, the upper surface of the transmission shaft is fixedly connected to the shunt shaft, the side wall of the transmission shaft is sleeved with a guide bushing, and the surrounding of the guide bush is respectively connected to the side of the shunt conduit through a connecting rod. The wall is fixedly connected, and the lower surface of the transmission shaft is fixedly connected with the central liquid level float ball through bolts. The side wall is adapted to the limit groove, and the limit groove is connected with the side liquid level float ball by nesting and buckle.

Further, the side liquid level float ball and the diversion conduit are respectively arranged in a staggered manner, and the diversion conduit is provided with a diversion port on the lower surface, and the diversion port is inclined.

Compared with the prior art, the beneficial effects of the present invention are:

1. a kind of domestication method and equipment of Bacillus subtilis for improving the bottom of aquatic products proposed by the present invention, by preparing culture medium, Bacillus subtilis is activated in the culture medium, for further the mechanism of domestication of Bacillus subtilis in aquaculture wastewater is provided. A good environment enables the medium to achieve the effect of high-density growth of organic matter in aquaculture wastewater, and has a specific cultivation effect on Bacillus, and gradually domesticates to obtain Bacillus strains that are specifically adapted to the environment at the bottom of aquaculture ponds , non-toxic and harmless, environmentally friendly, can more effectively oxidize the organic matter in the bottom mud, thereby improving the bottom environment of the pond, with strong pertinence.

2. A kind of domestication method and equipment of Bacillus subtilis for aquaculture bottom improvement proposed by the present invention, by adding the configured simulated aquaculture wastewater into the hydraulic tank, the scale is convenient to observe the added amount of the simulated aquaculture wastewater, which plays an important role. The effect of quantitative addition, the scale is accurate, so as to ensure the accuracy of the required dose, improve the accuracy of the addition amount, and reduce errors.

3. A kind of domestication method and equipment of Bacillus subtilis for aquaculture bottom improvement proposed by the present invention simulates that aquaculture waste water is transported to one side of the shunt fan blade through the liquid outlet, and drives the shunt shaft to rotate, so that the shunt conduit rotates at the same time. The simulated aquaculture wastewater is transported into the petri dish respectively, which improves the uniformity of the added wastewater, ensures the uniform mixing of the simulated aquaculture wastewater and the culture medium, and improves the domestication effect. During the up and down translation, the guide column effectively plays a guiding role, avoiding the inclination during the up and down movement, and avoiding the contamination caused by the contact between the diversion port at the lower end of the shunt conduit and the culture medium.

Description of drawings

Fig. 1 is the domestication method flow chart of Bacillus subtilis of the present invention;

Fig. 2 is the axonometric view of the structure of the incubator of the present invention;

Fig. 3 is the axonometric view of the internal structure of the incubator of the present invention;

Fig. 4 is the axonometric view of the connection between the culture dish and the shunt conduit of the present invention;

5 is an exploded axonometric view of the connection between the lower liquid inlet pipe and the shunt box of the present invention;

FIG. 6 is a partial structure axonometric view of the present invention.

In the figure: 1. Incubator; 11. Upper liquid inlet pipe; 12. Lower liquid inlet pipe; 13. Guide column; 14. Support plate; 15. Liquid outlet; 2. Hydraulic bucket; 3. Petri dish; 31. Heating wire; 4. shunt box; 41, shunt conduit; 42, shunt shaft; 43, shunt fan blade; 5, drive shaft; 51, guide bushing; 52, connecting rod; 53, center liquid level float; 54, Floating plate; 55, side liquid level float; 56, limit groove.

Detailed ways

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, but not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

Please refer to Fig. 1, a domestication method of Bacillus subtilis for improving the bottom of aquatic products, including the following steps:

S1: activate Bacillus, prepare a culture medium, and activate Bacillus subtilis in the culture medium;

Described culture medium is made up of following raw material components and weight parts: the consumption of distilled water is 1L, the consumption of glucose is 20g, the consumption of peptone is 15g, the consumption of sodium chloride is 5g, the consumption of beef extract is 0.5g, the consumption of agar is 0.5g. The dosage is 20g, the pH value of the culture medium is 7, and the temperature is 32 degrees Celsius. The culture medium contains bacterial species factors with high specificity and strong representation to the environment of aquaculture wastewater, reducing other factors that are not related to the environment of aquaculture wastewater. The interference of large factors provides a good environment for further domesticating the mechanism of Bacillus in aquaculture wastewater;

S2: domesticate Bacillus, prepare simulated aquaculture wastewater, and gradually increase the content of simulated aquaculture wastewater into the culture medium;

S201: take f/2 medium as the base liquid, add ammonium chloride and sodium dihydrogen phosphate, so that the initial PO ₄ ^3- -P(NaH ₂ PO ₄ ) concentration of simulated aquaculture wastewater is 1.5 mg/L, NH ₄ +- The concentration of N(NH ₄ Cl) was 0mg/L (f/2 medium), 20mg/L, 40mg/L, 80mg/L, 100mg/L, and the concentration of COD (glucose) was 500mg/L;

The simulated aquaculture wastewater is added with trace element liquid in the preparation process, and the trace element liquid is composed of the following raw material components and parts by weight: the dosage of H ₃ BO ₃ is 2.9*10 ^-3 g/L, MnCI ₂ -4H ₂ O The dosage of ZnSO 4 -7H 2 O is 1.8*10 ^-5 g/L, the dosage of ZnSO ₄ -7H ₂ O is 2.2*10 ^-4 g/L, the dosage of Na ₂ MoO ₄ -2H ₂ O is 3.9*10 ^-4 g/L, The dosage of CuSO ₄ -5H ₂ O is 8.0*10 ^-5 g/L, the dosage of Co(NO ₃ ) ₂ -6H ₂ O is 5.0*10 ^-5 g/L, and the composition of simulated aquaculture wastewater by adding trace element solution Similar to the current aquaculture wastewater, the medium can achieve the effect of high-density growth of organic matter in aquaculture wastewater, controllably promote the growth of certain organic matter, and can study the tendency of different organic matter to the same strain, It has a specific culture effect on Bacillus;

S3: Obtain Bacillus, gradually domesticate to obtain Bacillus strains that are specifically adapted to the environment at the bottom of aquaculture ponds, non-toxic and harmless, environmentally friendly, and can more effectively oxidize the organic matter in the sediment, thereby improving the pond. The bottom environment is highly targeted, and different Bacillus strains are domesticated according to the bottom environment of different aquaculture ponds.

Please refer to Fig. 2-4, an equipment for the domestication method of Bacillus subtilis for improving the bottom of aquatic products, including an incubator 1 for placing culture medium and preparing simulated aquaculture wastewater, the center of the upper surface of the incubator 1 is connected to a hydraulic The barrel 2 is fixedly connected, the hydraulic barrel 2 is set as a transparent glass cylinder, the side wall of the hydraulic barrel 2 is provided with a scale, the lower surface of the inner wall of the incubator 1 is fixedly connected to the culture dish 3 through bolts, and the center of the upper surface of the inner wall of the incubator 1 is through bolts It is fixedly connected with the upper liquid inlet pipe 11, the lower end of the upper liquid inlet pipe 11 is nested and connected with the lower liquid inlet pipe 12, the upper surface of the inner wall of the incubator 1 is fixedly connected to the guide column 13 through bolts, and the lower surface of the guide column 13 is connected to the support plate through bolts. 14 is fixed around the connection, the guide column 13 is set as a telescopic column, the center of the support plate 14 is fixedly connected with the outer wall of the lower liquid inlet pipe 12, the outer wall of the culture dish 3 is covered with a heating wire 31, the culture dish 3 is equipped with a culture medium, and the heating wire 31 is effective. To ensure that the temperature of the liquid in the petri dish 3 is stably maintained at the required temperature, by adding the configured simulated aquaculture wastewater to the hydraulic tank 2, the hydraulic tank 2 will pressurize the simulated aquaculture wastewater to the upper liquid inlet pipe 11 and the lower inlet. In the liquid pipe 12, the scale is convenient to observe the added amount of simulated aquaculture wastewater, which has the effect of quantitative addition, and the scale is accurate, so as to ensure the accuracy of the required dose, improve the accuracy of the added amount, and reduce errors. The liquid level in the pan 3 moves up and down, and the guide column 13 effectively plays a guiding role to avoid tilting during the up and down movement;

5-6, the lower surface of the lower liquid inlet pipe 12 is rotatably connected to the shunt box 4 through a bearing, the shunt box 4 is fixedly connected to the shunt conduit 41 through bolts, and the lower surface of the inner wall of the shunt box 4 rotates through the bearing and the shunt shaft 42 Connection, the side wall of the shunt shaft 42 is respectively provided with evenly distributed shunt fan blades 43, the lower surface of the lower liquid inlet pipe 12 is respectively provided with a liquid outlet 15, the liquid outlet 15 is respectively inclined and is respectively below the lower surface of the liquid inlet pipe 12 The center of the circle is symmetrically distributed and inclined in the same direction. The center of the lower surface of the shunt box 4 is rotatably connected with the transmission shaft 5 through the bearing. The upper surface of the transmission shaft 5 is fixedly connected with the shunt shaft 42. The periphery of the shaft sleeve 51 is fixedly connected to the side wall of the shunt conduit 41 through the connecting rod 52, and the lower surface of the transmission shaft 5 is fixedly connected to the center liquid level float ball 53 through bolts. The level float ball 55 is fixedly connected, and one end of the floating plate 54 is respectively provided with a limit groove 56 which is adapted to the side wall of the side liquid level float ball 55. The liquid level float 55 and the shunt duct 41 are respectively arranged in a staggered manner. The lower surface of the shunt duct 41 is provided with a diversion port, and the diversion port is arranged at an inclination. The simulated aquaculture wastewater is transported to the side of the shunt fan blade 43 through the liquid outlet 15. And drive the shunt shaft 42 to rotate, so that the transmission shaft 5 drives the shunt conduit 41 to rotate in the petri dish 3 through the guide bushing 51, and the shunt conduit 41 transfers the simulated aquaculture wastewater to the petri dish 3 while rotating, and the simulated aquaculture wastewater is along the culture dish 3. The inner wall of the dish 3 flows into the culture medium, and the center liquid level float ball 53, the floating plate 54 and the side liquid level float ball 55 adjust the height of the shunt duct 41 according to the height of the liquid level in the culture dish 3, so as to avoid the diversion port at the lower end of the shunt duct 41 and the culture medium. Contact causes contamination.

To sum up, the domestication method and equipment of Bacillus subtilis used for improving the bottom of aquatic products, by preparing the culture medium and activating Bacillus subtilis in the culture medium, provides a good mechanism for further domesticating Bacillus subtilis in aquaculture wastewater. environment, so that the medium can achieve the effect of high-density growth of organic matter in aquaculture wastewater, and has a specific cultivation effect on Bacillus, and gradually domesticate to obtain Bacillus strains that are specifically adapted to the environment at the bottom of aquaculture ponds. It is harmless and environmentally friendly, and can more effectively oxidize the organic matter in the sediment, thereby improving the bottom environment of the pond. It is convenient to observe the added amount of simulated aquaculture wastewater, which has the effect of quantitative addition, and the scale is accurate, so as to ensure the accuracy of the required dose, improve the accuracy of the added amount, and reduce errors. The simulated aquaculture wastewater is transported to the diversion through the liquid outlet 15 One side of the fan blade 43, and drives the shunt shaft 42 to rotate, so that the shunt conduit 41 transports the simulated aquaculture wastewater to the petri dish 3 while rotating, and the simulated aquaculture wastewater flows into the culture medium along the inner wall of the petri dish 3, and the center liquid level. The floating ball 53 , the floating plate 54 and the side liquid level floating ball 55 adjust the height of the shunt duct 41 according to the liquid level in the petri dish 3 , the support plate 14 moves up and down with the liquid level in the petri dish 3 , and the guide column 13 effectively plays the role of The guiding function avoids the inclination during the up and down movement, and avoids the contamination caused by the contact between the diversion port at the lower end of the shunt conduit 41 and the culture medium.

The above is only a preferred embodiment of the present invention, but the protection scope of the present invention is not limited to this. The equivalent replacement or change of the inventive concept thereof shall be included within the protection scope of the present invention.

Claims (9)

1. A domestication method of bacillus subtilis for changing bottom of aquatic products is characterized by comprising the following steps: the method comprises the following steps:

s1: activating bacillus, preparing a culture medium, and activating the bacillus subtilis in the culture medium;

s2: domesticating bacillus, preparing simulated aquaculture wastewater, and gradually increasing the content of the simulated aquaculture wastewater in a culture medium;

s3: obtaining bacillus, gradually domesticating to obtain bacillus strain specifically adapted to the bottom environment of the aquaculture pond.

2. The domestication method of bacillus subtilis for improving the bottom of aquatic products as claimed in claim 1, wherein: in S1, the culture medium consists of the following raw material components in parts by weight: the using amount of distilled water is 1L, the using amount of glucose is 20g, the using amount of peptone is 15g, the using amount of sodium chloride is 5g, the using amount of beef extract is 0.5g, and the using amount of agar is 20 g.

3. The domestication method of bacillus subtilis for improving the bottom of aquatic products as claimed in claim 2, wherein: for S1, the ph of the medium was 7 and the temperature was 32 degrees celsius.

4. The domestication method of bacillus subtilis for improving the bottom of aquatic products as claimed in claim 1, wherein: aiming at S2, the preparation of the simulated aquaculture wastewater comprises the following steps:

s201: taking f/2 culture medium as base liquid, adding ammonium chloride and sodium dihydrogen phosphate to simulate initial PO of the aquaculture wastewater ₄ ^3- -P(NaH ₂ PO ₄ ) Concentration 1.5mg/L, NH ₄ +-N(NH ₄ Cl) concentrations were 0mg/L (f/2 medium), 20mg/L, 40mg/L, 80mg/L, 100mg/L, respectively, and COD (glucose) concentration was 500 mg/L.

5. The domestication method of bacillus subtilis for improving the bottom of aquatic products as claimed in claim 4, wherein: aiming at S201, trace element liquid is added into the simulated aquaculture wastewater in the preparation process, and the trace element liquid comprises the following raw material components in parts by weight: h ₃ BO ₃ In an amount of 2.9 x 10 ^-3 g/L，MnCI ₂ -4H ₂ The amount of O is 1.8 x 10 ^-5 g/L，ZnSO ₄ -7H ₂ The amount of O is 2.2 x 10 ^-4 g/L，Na ₂ MoO ₄ -2H ₂ The amount of O is 3.9 x 10 ^-4 g/L，CuSO ₄ -5H ₂ The amount of O is 8.0 x 10 ^-5 g/L，Co(NO ₃ ) ₂ -6H ₂ The amount of O is 5.0 x 10 ^-5 g/L。

6. The apparatus for acclimatization method of bacillus subtilis for aquaculture according to claim 1, comprising an incubator (1) for placing culture medium and dispensing simulated aquaculture wastewater, characterized in that: incubator (1) upper surface center passes through bolt and liquid feeding pressure bucket (2) fixed connection, liquid feeding pressure bucket (2) set up to transparent glass section of thick bamboo, liquid feeding pressure bucket (2) lateral wall is provided with the scale, incubator (1) inner wall lower surface passes through bolt and culture dish (3) fixed connection, incubator (1) inner wall upper surface center passes through bolt and last feed pipe (11) fixed connection, go up feed pipe (11) lower extreme and lower feed pipe (12) nested connection, incubator (1) inner wall upper surface is all around respectively through bolt and guide post (13) fixed connection, guide post (13) lower surface passes through bolt and fagging (14) fixed connection all around, guide post (13) set up to flexible post, fagging (14) center and lower feed pipe (12) outer wall fixed connection, culture dish (3) outer wall cover has heater strip (31).

7. The domestication method and equipment of bacillus subtilis for improving bottom of aquatic products as claimed in claim 6, wherein: lower feed liquor pipe (12) lower surface passes through the bearing and divides flow box (4) to rotate to be connected, divide flow box (4) to rotate through bolt and reposition of redundant personnel pipe (41) fixed connection all around respectively, divide flow box (4) inner wall lower surface to pass through the bearing and divide flow shaft (42) to rotate to be connected, divide flow shaft (42) lateral wall to be provided with evenly distributed's reposition of redundant personnel flabellum (43) respectively, lower feed liquor pipe (12) lower surface is provided with liquid outlet (15) respectively, liquid outlet (15) are the slope form respectively and below feed liquor pipe (12) lower surface centre of a circle central symmetry distributes and slope setting to same direction respectively.

8. The domestication method and equipment of bacillus subtilis for improving bottom of aquatic products as claimed in claim 7, wherein: the center of the lower surface of the flow distribution box (4) is rotatably connected with a transmission shaft (5) through a bearing, the upper surface of the transmission shaft (5) is fixedly connected with a flow distribution shaft (42), a guide shaft sleeve (51) is sleeved on the side wall of the transmission shaft (5), the periphery of the guide shaft sleeve (51) is fixedly connected with the side wall of a flow distribution guide pipe (41) through a connecting rod (52), the lower surface of the transmission shaft (5) is fixedly connected with a central liquid level floating ball (53) through a bolt, the periphery of the central liquid level floating ball (53) is fixedly connected with a side liquid level floating ball (55) through a floating plate (54), one end of the floating plate (54) is provided with a limiting groove (56) matched with the side wall of the side liquid level floating ball (55), and the limiting groove (56) is connected with the side liquid level floating ball (55) in a nested and buckled mode.

9. The domestication method and equipment of bacillus subtilis for improving bottom of aquatic products as claimed in claim 8, wherein: the side liquid level floating balls (55) and the shunt pipes (41) are arranged in a staggered mode respectively, and the lower surfaces of the shunt pipes (41) are provided with flow guide openings which are arranged in an inclined mode.

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