CN111410303A - Rapid formation of granular sludge coupled with denitrification anaerobic methane oxidation and anammox - Google Patents

Abstract

A method for rapid formation of denitrifying anaerobic methane oxidation coupled with anammox granule sludge, wherein anammox granular sludge is used to rapidly adsorb flocculent denitrifying anaerobic methane oxidizing microorganisms, and 3-oxo- C6‑HSL and DPD, to promote the colonization of anaerobic methane oxidizing microorganisms on the surface of anammox granular sludge, and then through continuous flow cultivation to ensure the presence of sufficient dissolved _CH4 , using high concentrations of ammonia nitrogen and nitrate to strengthen Rapid growth and metabolic activity of denitrifying anaerobic methane oxidizing microorganisms, and adding 3‑oxo‑C6‑HSL, C6‑HSL, C12‑HSL and DPD to enhance the rapid growth of anaerobic methane oxidizing archaea and bacteria, adding Ca ²⁺ and Sr ²⁺ mixed ions enhance the role of the second messenger of anaerobic methanotrophic archaea, promote the rapid growth and film formation of anaerobic methanotrophic archaea, and strengthen the stability and particle strength of coupled particles, thereby promoting the formation of coupled particles. Form quickly.

Description

Rapid formation of granular sludge coupled with denitrification anaerobic methane oxidation and anaerobic ammonia oxidation

technical field

The invention belongs to the technical field of sewage treatment, and in particular relates to a method for rapidly forming granular sludge coupled with denitrification anaerobic methane oxidation and anaerobic ammonia oxidation.

Background technique

With the development of people's production and life, a large amount of nitrogen-containing wastewater is produced, and if it is not treated and discharged directly, it will cause serious harm to the environment. Nitrogen-containing substances will not only cause eutrophication of water bodies and seriously damage aquatic ecosystems; after entering groundwater, excessive intake of NO ₃ ^- or NO ₂ ^- will also cause "blue baby disease". The existing denitrification technologies mainly include physical method, chemical method and biological method. Compared with the former two methods, biological denitrification technology has the advantages of mild conditions, low treatment cost and high denitrification efficiency, which is favored by people. .

Among the many biological denitrification technologies, anammox technology is known as one of the most economical denitrification technologies due to its advantages such as no aeration, no need to add organic matter, and less excess sludge output. The technology can simultaneously convert nitrite and ammonia nitrogen into N ₂ under anaerobic conditions, and has the advantages of low operating cost, economical efficiency, and no secondary pollutants. However, according to the reaction mechanism of anammox, the anammox reaction will convert 21% of nitrite nitrogen into nitrate nitrogen, so that the theoretical total nitrogen removal rate is only about 80%; The ratio of water nitrite and ammonia nitrogen is not ideal, and the total nitrogen removal rate can only be maintained at about 70%.

Denitrifying anaerobic methane oxidizing microorganisms discovered in recent years include denitrifying anaerobic methane oxidizing archaea and denitrifying anaerobic methane oxidizing bacteria _. anaerobic methane-oxidizing bacteria can convert nitrite to nitrogen using _CH4 as an electron donor. The synergistic effect of denitrifying anaerobic methane oxidizing microorganisms and anammox bacteria can not only effectively convert nitrate nitrogen generated by anammox reaction, but also improve its total nitrogen removal rate; denitrifying anaerobic methane oxidizing bacteria can also effectively Remove nitrite nitrogen in influent, adjust the ratio of ammonia nitrogen and nitrite nitrogen in influent, and promote anaerobic ammonium oxidation; at the same time, denitrifying anaerobic methane oxidizing microorganisms can also remove soluble CH in influent water _4. It can simultaneously remove the greenhouse gas methane and nitrogen-containing pollutants.

However, denitrifying anaerobic methane-oxidizing archaea, denitrifying anaerobic methane oxidizing bacteria and anammox bacteria are all autotrophic bacteria, which have the disadvantages of slow growth and long doubling time. The synergistic state of oxidizing microorganisms and anammox bacteria mostly exists in the form of flocculent sludge, with poor sedimentation effect and easy loss of functional microorganisms. In contrast, the existence of granular sludge has the advantages of high sludge concentration, fast sedimentation, and high stability. application potential.

In patent 201711472212.X, Xie Guojun et al. proposed a new type of membrane aerated anaerobic granular sludge reactor and its high-efficiency denitrification and greenhouse gas emission reduction method, which solved the problem that the solubility of gaseous substances in aqueous solutions is very low, and gas-liquid transmission The problem of large mass resistance and low mass transfer rate. In the method, flocculent sludge and anaerobic granular sludge containing denitrifying anaerobic methane oxidizing bacteria, denitrifying anaerobic methane oxidizing archaea and anammox functional microorganisms after enrichment culture are used as inoculated sludge. The membrane aerated anaerobic granular sludge reactor will form granular sludge rich in denitrifying anaerobic methane oxidizing bacteria and anaerobic ammonia oxidizing bacteria after 100 days of operation. The granular sludge is round or ellipsoid with a particle size of 500-1000um .

CN101058463A discloses a wastewater biological treatment method realizing the coupling of anammox and methanation and denitrification, inoculating granular sewage with methanation, denitrification and anammox activities in an expanded granular sludge bed (EGSB) reactor Sludge, inoculate aerobic ammonium oxidizing bacteria in biological aerated filter (BAF) or SHARON process or SBR reactor, combine expanded granular sludge bed (EGSB) reactor with biological aerated filter (BAF) or SHARON process or The SBR reactors are connected to each other, and by reasonably controlling pH value, temperature, dissolved oxygen, external reflux ratio, loading rate, redox potential, COD concentration of organic matter, and ammonia nitrogen concentration, the wastewater with the coupling of anaerobic ammonia oxidation and methanation and denitrification has been successfully realized. biological treatment. The technology can simultaneously remove organic COD and inorganic ammonia components at high volume loading rate, while saving a considerable part of energy and organic carbon source consumption, and can be applied to the biological treatment of general urban sewage and industrial wastewater.

SUMMARY OF THE INVENTION

Based on this, the present invention proposes a rapid formation method of denitrifying anaerobic methane oxidation and anammox coupled granular sludge, which can not only greatly shorten the cultivation of denitrifying anaerobic methane oxidation and anammox coupled granular sludge It can also effectively solve the problems of poor sludge settling effect and easy loss of functional microorganisms in the existing coupling technology of denitrifying anaerobic methane oxidizing microorganisms and anammox bacteria.

Based on one aspect of the present invention, a method for rapidly forming granular sludge coupled with denitrification anaerobic methane oxidation and anammox ammonium oxidation is disclosed, comprising the following steps: (1) using an up-flow anaerobic sludge bed reactor, The flocculent sludge containing denitrifying anaerobic methane oxidizing bacteria and denitrifying anaerobic methane oxidizing archaea and the mixed sludge composed of anammox granular sludge after enrichment culture were used as inoculum, and the temperature was controlled at 25 -35°C, the upward flow rate of the reactor is 3-5m/h, intermittently add nitrate concentrate, nitrite concentrate and ammonia nitrogen concentrate for cultivation, and ensure that the nitrite concentration in the system is maintained below 20mgN/L, Ammonia nitrogen and nitrate concentrations are both below 100mgN/L; intermittent exposure to _CH4 gas ensures that the dissolved methane concentration in the system is above 10mg/L; at the same time, 1-5μmol/L of 3-oxo-C6-HSL (N- 3-oxo-hexanoyl homoserine lactone) and 10-50ng/L DPD (4,5-dihydroxy-2,3-pentanedione) to promote denitrifying anaerobic methane oxidizing microorganisms in anammox Colonization on the surface of granular sludge; enter into step (2) after intermittent cultivation for 3-14 days; (2) continuous inflow water, the influent water contains soluble CH ₄ , nitrate and ammonia nitrogen, and the concentration of the CH ₄ is 15～ 20mg/L, the nitrate concentration is kept at 100-800mgN/L, the ammonia nitrogen concentration is 100-800mgN/L, the water retention time is kept at 4h-24h, and the concentration of 3-oxo-C6- HSL (N-3-oxo-hexanoyl homoserine lactone), C6-HSL (N-hexanoyl-L-homoserine lactone) and C12-HSL (N-dodecanoyl-L-homoserine lactone) ), and DPD of 20-100ng/L, adding mixed ions of Ca ²⁺ and Sr ²⁺ at the same time, the molar ratio of the two is 1:1-2:1, and the concentration is 2-5μmol/L to improve denitrification. The anaerobic methane archaeal activity promotes its rapid growth and film formation, and promotes the rapid formation of coupled granular sludge, and continues to cultivate for 30-90 days to complete the coupled granular sludge cultivation.

The intermittent addition of nitrate concentrate, nitrite concentrate and ammonia nitrogen concentrate culture described in the step (1) is carried out according to the following steps: when the nitrite concentration in the system is below 5mg/L, the nitrate and ammonia nitrogen concentrations are low At 10 mg/L, the nitrite concentrate, nitrate concentrate and ammonia nitrogen concentrate were injected with a syringe pulse respectively.

As described in step (1), the mass ratio of the flocculent sludge containing denitrifying anaerobic methane oxidizing bacteria and denitrifying anaerobic methane oxidizing archaea and anammox granular sludge after enrichment culture is 1 :1-5:1-3;

In step (1), the rising flow rate of the reactor is preferably 2.5-3.0, more preferably 2.8;

In step (2), the concentrations of C3-oxo-C6-HSL, C6-HSL, C12-HSL and DPD are preferably 3-4 μmol/L, 2-3 μmol/L, 3-4 μmol/L and 40-60ng/L, more Preferably it is 3.5μmol/L, 2.5μmol/L, 3.5μmol/L and 50ng/L;

In the step (2), the preferred dosing method of the mixed ions of Ca ²⁺ and Sr ²⁺ is as follows: the molar ratio of Ca ²⁺ and Sr ²⁺ is 1:1.5-3 at the initial stage of continuous flow startup, and the increase of EPS reaches 40%. , changed to Ca ²⁺ and Sr ²⁺ molar ratio of 1:0.5-1.5.

Another aspect of the present invention discloses a granular sludge cultured by using the method for rapidly forming granular sludge by coupling anaerobic methane oxidation with denitrification and anammox, wherein the particle size of the granular sludge is 0.4~ 2.0mm; the particles are round or ellipsoid, with extracellular polymers distributed on the surface of the particles; denitrifying anaerobic methane archaea and denitrifying anaerobic methane oxidizing bacteria are located outside the granular sludge, and anammox bacteria are located in the granular sludge internal.

The main denitrifying anaerobic methane oxidizing archaea is Candidatus Methanoperedens; the main denitrifying anaerobic methane oxidizing bacterium is Candidatus Methylomirabilis; the main anammox bacterium is Candidatus Brocadia;

The final nitrate consumption rate of granular sludge is 50-150mgN/L/d; the consumption rate of ammonia nitrogen is 200-1000mgN/L/d; the denitrification load is 500-2000mgN/L/d; the granular sludge is in the reactor The concentration in 5-50gVSS/L.

The method adopts the mixed sludge of activated sludge composed of denitrifying anaerobic methane oxidation microorganisms and anaerobic ammonia oxidation granular sludge as the inoculum, and simultaneously cultivates intermittently, and utilizes anaerobic ammonia oxidizing bacteria to oxidize denitrifying anaerobic methane. Ecological compatibility of microorganisms, rapid adsorption of denitrifying anaerobic methane oxidizing microorganisms, avoiding the loss of functional microorganisms; adding signal molecules 3-oxo-C6- HSL and DPD promote the colonization of anaerobic methane oxidizing microorganisms on the surface of anammox granular sludge to form a preliminary coupled granular sludge prototype; during continuous flow operation, when sufficient dissolved CH ₄ is ensured, high The concentration of ammonia nitrogen and nitrate can enhance the rapid growth and metabolic activity of denitrifying anaerobic methane oxidizing archaea; adding signal molecules 3-oxo-C6-HSL, C6-HSL, C12-HSL and DPD can enhance anaerobic methane oxidation Rapid growth of archaea; adding mixed ions of Ca ²⁺ and Sr ²⁺ at the same time to strengthen the role of the second messenger of anaerobic methane oxidizing archaea and promote the rapid growth and film formation of anaerobic methane oxidizing archaea; Ca ²⁺ and Sr The addition of ²⁺ can also promote the production of EPS and strengthen the stability and strength of coupled particles, thereby promoting the rapid formation of coupled particles.

Beneficial technical effects of the present invention: the present invention can quickly form coupled granular sludge, the formation time of the coupled granular sludge prototype is only about 7 days, and the formation time of mature coupled granular sludge is only about 50 days; the stability of the obtained coupled granular sludge is Strong, high sludge concentration, good coupling granular sludge treatment effect, the final consumption rate of nitrate and ammonia nitrogen has reached a high level, the denitrification load can reach up to 1250mgN/L/d; the particle size is moderate and evenly distributed ; Coupled granular sludge can also make full use of the methane gas generated in the sewage treatment plant, and can simultaneously remove the greenhouse gas methane and nitrogen-containing pollutants.

Description of drawings

Figure 1 is an optical photograph of the coupled granular sludge cultivated in Example 8;

FIG. 2 is an optical micrograph of the coupled granular sludge cultured in Example 8. FIG.

Detailed ways

The technical solutions of the present invention are not limited to the specific embodiments listed below, but also include any combination of specific embodiments.

Example 1

The rapid formation method of denitrifying anaerobic methane oxidation and anaerobic ammonium oxidation coupled granular sludge is carried out according to the following methods: (1) an up-flow anaerobic sludge bed reactor is used, and the enriched culture containing denitrifying anaerobic sludge is used. The mixed sludge composed of oxygen methane oxidizing bacteria, denitrifying anaerobic methane oxidizing archaeal flocculent sludge, and anaerobic ammonia oxidation granular sludge is used as the inoculum. /h, intermittently add nitrate concentrate, nitrite concentrate and ammonia nitrogen concentrate for cultivation, and ensure that the nitrite concentration in the system is maintained below 20mgN/L, and the ammonia nitrogen and nitrate concentrations are both below 100mgN/L; The aeration head is used to aerate CH ₄ gas intermittently to ensure that the dissolved methane concentration in the system is above 10mg/L; at the same time, 2μmol/L of 3-oxo-C6-HSL and 20ng/L of DPD are added to promote anaerobic methane oxidation The colonization of microorganisms on the surface of ANAMMOX granular sludge; the batch culture ended after 7 days; (2) continuous inflow of water, the concentration of soluble _CH4 in the influent water was 15-20mg/L, and the nitrate concentration was maintained at 530mgN/L, The concentration of ammonia nitrogen was 500mgN/L, the water retention time was kept at 4h-24h, and the concentrations of 3-oxo-C6-HSL, C6-HSL and C12-HSL were 2-5μmol/L, and 20-100ng/L. DPD, adding mixed ions of Ca ²⁺ and Sr ²⁺ at the same time, the molar ratio of the two is 1:1-2:1, and the concentration is 2-5μmol/L, in order to improve the denitrifying anaerobic methane archaea activity and promote its Rapid growth and film formation promote the rapid formation of coupled granular sludge, and continue to cultivate for 50 days to complete coupled granular sludge cultivation.

Example 2

The difference between this embodiment and the specific embodiment 1 is that in step 1, the upward flow rate of the reactor is 3.5 m/h, and in step 2, the concentrations of C3-oxo-C6-HSL, C6-HSL, C12-HSL and DPD are 3.5 μmol/h L, 2.5μmol/L, 3.5μmol/L and 40ng/L. Other steps and parameters are the same as in the first embodiment.

Example 3

The difference between this embodiment and the specific embodiment 1 is: in the second step, the dosing method of the mixed ions of Ca ²⁺ and Sr ²⁺ is that the molar ratio of Ca ²⁺ and Sr ²⁺ is 1:2 in the initial stage of continuous flow startup, and the The EPS increase reached 40%, and the molar ratio of Ca ²⁺ and Sr ²⁺ was changed to 1:1. Other steps and parameters are the same as in the first embodiment.

Example 4

The difference between this embodiment and the specific embodiment 1 is: in step 1, the concentration of nitrite in the system is maintained at 10 mgN/L, and the concentrations of ammonia nitrogen and nitrate are both 100 mgN/L; in step ₂ , the concentration of soluble CH in the system is 18mg/L, ammonia nitrogen and nitrate concentrations are both 700mgN/L. Other steps and parameters are the same as in the first embodiment.

Example 5

In the first step, the upward flow rate of the reactor was 3.8 m/h, and in the second step, the concentrations of C3-oxo-C6-HSL, C6-HSL, C12-HSL and DPD were 3.8 μmol/L, 2.8 μmol/L, 3.5 μmol/L and 3.8 μmol/L. 60ng/L. Other steps and parameters are the same as in the first embodiment.

Example 6

In the first step, the nitrite concentration in the system was maintained at 15 mgN/L, and the ammonia nitrogen and nitrate concentrations were both at 80 mgN/L; in the second step, the soluble _CH4 concentration in the system was 19 mg/L. Other steps and parameters are the same as in the first embodiment.

Example 7

In the first step, the nitrite concentration in the system was maintained at 14 mgN/L; in the second step, the ammonia nitrogen and nitrate concentrations in the system were both 800 mgN/L. Other steps and parameters are the same as in the first embodiment.

Example 8

The rapid formation method of denitrifying anaerobic methane oxidation and anaerobic ammonium oxidation coupled granular sludge includes the following steps: (1) using an up-flow anaerobic sludge bed reactor to enrich and cultivate denitrifying anaerobic methane containing denitrifying anaerobic methane. The mixed sludge composed of oxidizing bacteria, denitrifying anaerobic methane oxidation archaeal flocculent sludge, and anaerobic ammonia oxidation granular sludge is used as the inoculum. Add nitrate concentrate, nitrite concentrate and ammonia nitrogen concentrate for cultivation, and ensure that the nitrite concentration in the system is maintained at 18mgN/L, and the ammonia nitrogen and nitrate concentrations are both 90mgN/L; intermittently exposing _CH4 gas, Ensure that the dissolved methane concentration in the system is 10mg/L; at the same time add 2μmol/L 3-oxo-C6-HSL and 20ng/L DPD to promote denitrifying anaerobic methane oxidation microorganisms in anammox granular sludge The colonization of the surface ends after 7 days of intermittent culture; (2) continuous inflow of water, the concentration of influent _CH4 is 17mg/L, the concentration of nitrate is kept at 530mgN/L, the concentration of ammonia nitrogen is 500mgN/L, and the water retention time is kept at At 12h, 3-oxo-C6-HSL, C6-HSL and C12-HSL with a concentration of 3.5μmol/L, and DPD of 40ng/L were added, and mixed ions of Ca ²⁺ and Sr ²⁺ were added at the same time. The molar ratio of the two is 2:1, and the concentrations are 4 μmol/L and 2 μmol/L, respectively, to improve the activity of denitrifying anaerobic methane archaea, promote their rapid growth and film formation, and promote the rapid formation of coupled granular sludge. Continue to cultivate for 50 Day to complete the coupled granular sludge culture.

After 50 days of culture, the granular sludge concentration was 15 gVSS/L, and the particle size of the coupled granular sludge was in the range of 0.4-2.0 mm; the particles were round or ellipsoid, and extracellular polymers were distributed on the surface of the particles; denitrifying anaerobic methane was ancient. The bacteria and denitrifying anaerobic methane oxidizing bacteria are located outside the granular sludge, and the anammox bacteria are located inside the granular sludge; the main denitrifying anaerobic methane archaea is Candidatus Methanoperedens; the main denitrifying anaerobic methane bacterium is Candidatus Methylomirabilis; The main anammox bacteria was Candidatus Brocadia; the final nitrate consumption rate of coupled particles was 95.2 mgN/L/d; the consumption rate of ammonia nitrogen was 650 mgN/L/d; the denitrification load reached 1250 mgN/L/d.

The above embodiments are illustrative of the present invention, rather than limiting the scope of protection of the present invention. Based on the described embodiments, all other technical solutions obtained by those of ordinary skill in the art without creative work fall within the protection scope of the present invention.

Claims (9)

1. A method for quickly forming granular sludge by coupling denitrification anaerobic methane oxidation and anaerobic ammonia oxidation comprises (1) adopting an upflow anaerobic sludge bed reactor, taking mixed sludge formed by flocculent sludge containing denitrification anaerobic methane oxidation bacteria and denitrification anaerobic methane oxidation archaea and anaerobic ammonia oxidation granular sludge which are subjected to enrichment culture as an inoculum, controlling the temperature to be 25-35 ℃, controlling the rising flow rate of the reactor to be 3-5m/h, intermittently adding nitrate concentrated solution, nitrite concentrated solution and ammonia nitrogen concentrated solution for culture, ensuring that the concentration of nitrite in the system is kept below 20 mgN/L, the concentration of ammonia nitrogen and nitrate is below 100 mgN/L, and intermittently aerating CH₄Gas, ensuring the concentration of the dissolved methane in the system to be more than 10 mg/L, simultaneously adding 1-5 mu mol/L of 3-oxo-C6-HS L (N-3-oxo-hexanoyl homoserine lactone) and 10-50 ng/L of DPD (4, 5-dihydroxy-2, 3-pentanedione) to promote the permanent planting of denitrifying anaerobic methane oxidation microorganisms on the surface of anaerobic ammonia oxidation granular sludge, performing intermittent culture for 3-14 days, entering the step (2), and continuously feeding water in the step (2), wherein the fed water contains the dissolved CH₄Nitrate and ammonia nitrogen, the CH₄The concentration of the nitrate is 15-20 mg/L, the concentration of the nitrate is maintained at 100-800 mgN/L, the concentration of the ammonia nitrogen is 100-800 mgN/L, the water conservancy residence time is maintained at 4-24 h, and the adding concentration is 2-5 mu mo respectivelyl/L of 3-oxo-C6-HS L (N-3-oxo-hexanoyl homoserine lactone), C6-HS L (N-hexanoyl-L-homoserine lactone) and C12-HS L (N-dodecanoyl-L-homoserine lactone), and DPD of 20-100 ng/L, and adding Ca at the same time²⁺And Sr²⁺The molar ratio of the mixed ions to the mixed ions is 1:1-2:1, the concentration is 2-5 mu mol/L, so that the activity of the denitrifying anaerobic methane-oxidizing archaea is improved, the rapid growth and film formation of the denitrifying anaerobic methane-oxidizing archaea are promoted, the rapid formation of the coupled granular sludge is promoted, and the coupled granular sludge is cultured for 30-90 days to finish the culture of the coupled granular sludge.

2. The process according to claim 1, wherein the reactor upflow rate in step (1) is in the range of 2.5 to 3.0, more preferably 2.8.

3. The method according to claim 1, wherein the intermittently fed nitrate concentrated solution, nitrite concentrated solution and ammonia nitrogen concentrated solution in the step (1) are cultured by injecting the nitrite concentrated solution, the nitrate concentrated solution and the ammonia nitrogen concentrated solution respectively by using an injector in a pulse manner when the concentration of the nitrite in the system is below 5 mg/L and the concentration of the nitrate and the ammonia nitrogen is below 10 mg/L.

4. The process according to claim 1, wherein the concentrations of C3-oxo-C6-HS L, C6-HS L, C12-HS L and DPD in step (2) are preferably 3-4. mu. mol/L, 2-3. mu. mol/L, 3-4. mu. mol/L and 40-60 ng/L.

5. The method according to claim 1, wherein Ca is contained in the step (2)²⁺And Sr²⁺The adding mode of the mixed ions is that Ca is adopted at the initial stage of continuous flow starting²⁺And Sr²⁺The molar ratio is 1:1.5-3, and Ca is changed when the EPS increase reaches 40 percent²⁺And Sr²⁺The molar ratio is 1: 0.5-1.5.

6. The granular sludge cultured by the method according to any one of claims 1 to 5, having a particle size of 0.4 to 2.0 mm; the particles are round or ellipsoidal, and extracellular polymers are distributed on the surfaces of the particles; the denitrifying anaerobic methane archaea and the denitrifying anaerobic methane oxidizing bacteria are positioned outside the granular sludge, and the anaerobic ammonia oxidizing bacteria are positioned inside the granular sludge.

7. The granular sludge of claim 6 wherein the predominant denitrifying anaerobic methanogen archaea is Candidatus Methanoperedens; the main denitrifying anaerobic methane-oxidizing bacteria are Candidatus methylomirabilis; the major anammox bacterium is Candidatus Brocadia.

8. The granular sludge as set forth in claim 6, characterized in that the final nitrate consumption rate of the granular sludge is 50-150 mgN/L/d, the ammonia nitrogen consumption rate is 200-1000 mgN/L/d, and the denitrification load is 500-2000 mgN/L/d.

9. The granular sludge as set forth in claim 6, characterized in that the concentration of the granular sludge in the reactor is 5-50 gVSS/L.

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