CN103074286B - High-salt heterotrophic nitrification-aerobic denitrification dephosphorization salinivibrio and application of salinivibrio in wastewater treatment - Google Patents
High-salt heterotrophic nitrification-aerobic denitrification dephosphorization salinivibrio and application of salinivibrio in wastewater treatment Download PDFInfo
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Abstract
The invention relates to high-salt heterotrophic nitrification-aerobic denitrification dephosphorization salinivibrio and an application of the salinivibrio in wastewater treatment. The salinivibrio has high endurance capacity to a high-salt environment, grows well under a high-salt condition, can metabolize by utilizing organic carbon as a sole carbon source and ammonia nitrogen as a sole nitrogen source, directly converts ammonia nitrogen into a gas product through the heterotrophic nitrification-aerobic denitrification action, and achieves denitrification; and the salinivibrio can also take nitrate nitrogen as a sole nitrogen source, converts nitrate nitrogen into a gas product through the aerobic denitrification action, and can take inorganic phosphorus in under an aerobic condition to convert into a self-component so as to remove phosphorus from sewage. The Salinivibrio is applicable to high-salt wastewater treatment, can achieve synchronous removal of nitrogen and phosphorus under a single aerobic condition, and effectively achieves biological denitrification and dephosphorization under the high-salt condition.
Description
Technical field
The present invention relates to a strain salt vibrios (Salinivibrio) and the application in high-salt wastewater is processed thereof.This bacterial strain has the function of heterotrophic nitrification-aerobic denitrification, can under high salt condition, realize the process of synchronous nitration denitrification denitrogenation, completes the removal that contains phosphor contaminant in sewage simultaneously.
Background technology
High-salt wastewater refers to that total salinity (with NaCl content meter) is at least 1% waste water, and these waste water, except the organism that contains a large amount of high densitys, also contain a large amount of inorganic salt, as Cl
-, Na
+, Ca
2+, SO
4 2-deng.Along with a large amount of discharges of direct seawater utilization and high saliferous trade effluent, the high-enriched organics in high-salt wastewater or nutrition, as COD, N, P etc., exert heavy pressures on to water body environment, aggravated the process of rivers and lakes eutrophication.Because high salinity can produce adverse influence to the eubolism of microorganism in conventional biological treatment system, mainly comprise: osmotic pressure is higher that microorganism cells plasmolysis makes growth be subject to hindering even dead; Microbial metabolism enzymic activity is obstructed; Water body density increases, and affects sludge settling effect etc.Therefore, under high salt condition, traditional biological dephosphorize denitrification technique often can not have been given play to good effect, and the denitrification dephosphorization technique of seeking under high salt condition is the emphasis of research always.
Traditional biological denitrogenation is, under the effect of microorganism, the organonitrogen in sewage and ammonia-state nitrogen are converted into N
2process, nitration reaction is to be completed by a group autotrophy aerobic microbiological, specifically divides two stages by Nitrosomas and two kinds of bacterium of Nitromonas, to be completed respectively.The nitration reaction of traditional biological denitrogenation need to be longer sludge age and aerobic condition, when large amount of organic exists, can cause the loss of nitrobacteria; Denitrifying bacterium needs shorter sludge age and anoxia condition, highly relies on organism and provides electron donor for its denitrification process.Because of the difference of nitrobacteria and denitrifying bacterium physiological mechanism caused tediously long based on this theoretical sewage water denitrification technical matters, energy consumption is large, floor space is large, to problems such as environmental change are responsive, nitric efficiency is not good.
The ultimate principle of traditional biological dephosphorization process is under anaerobic, PAOs produces ATP when decomposing cell poly-phosphate, and utilize ATP that the small organic molecules such as the voltaile fatty acid in environment are taken in born of the same parents, by its with PHB(poly--β hydroxybutyric acid salt) and the form of the organic granular such as glycogen be present in cell, the orthophosphoric acid salt (PO simultaneously poly-phosphate hydrolysis being generated
4 3-) discharge extracellular; Under aerobic condition, PAOs be take oxygen as electron acceptor(EA), and the energy that utilizes PHB metabolism to discharge absorbs and surpasses its needed phosphorus being stored in cell paste with the form of poly-phosphate of growing from sewage.By will being realized dephosphorization object with excess sludge discharge system by the phosphorus of bacterium excess ingestion.
It is exactly a contradictory entity that the difference of biological carbon and phosphorous removal mechanism has caused these two processes itself, and the basic reason that produces contradiction is that the needed optimal growth condition of the microorganism of difference in functionality is different.Nitrated need to be long nitrobacteria and the aerobic condition in mud age, denitrification needs short mud denitrifying bacteria and the anoxia condition in age, releasing phosphorus needs short mud polyP bacteria and the anaerobic condition in age, inhaling phosphorus needs aerobic condition.In addition between denitrifying bacteria and polyP bacteria, also exist because of the not enough competitive relation producing of carbon source in sanitary sewage.The development of biological denitrification phosphorous removal technique is mainly also to launch round the contradiction that realizes denitrogenation and dephosphorization in same Sewage treatment systems.Because of the existence of these intrinsic contradictions, caused that the most sewage works of China working cost is high, Nitrogen/Phosphorus Removal is unstable, compliance rate is lower.Under high salt condition, it is especially complicated and outstanding that the problem of dephosphorization denitrogenation becomes.Biological carbon and phosphorous removal research under non-high salt condition is existing greater advance in recent years, occurred the novel denitrification process such as SHARON, CANON, OLAND and ANNAMOX, but these techniques is not broken away from the restriction of the two-part biological denitrificaion of aerobic-anaerobic combination yet; Aspect biological phosphate-eliminating, found special Denitrifying Phosphate Accumulating Organisms (DPB), under the environment that can replace at hypoxic/anaerobic, realize the synchronous removal of nitrate nitrogen and phosphorus.Along with going deep into of research, investigator has also found special heterotrophic nitrification-aerobic denitrification bacterium, and this bacterioid can be realized the synchronous removal of ammonia nitrogen and total nitrogen under aerobic condition, has solved the contradiction of nitrification and denitrification.The microorganism of screening and find to have special denitrogenation dephosphorizing function is focus and the trend of biological carbon and phosphorous removal research field always.
The high-efficiency strain of Selection of Salt-Tolerant is also to solve the effective way of wastewater treatment under high salt condition and the focus that investigator pays close attention to.In the last few years, there is investigator to tame out halophilic bacterial classification by cultivation, and isolated Facultative Halophiles and halophilic bacterium from nature hypersaline environment, and be applied to high-salt wastewater processing and obtained good effect, as Woolard
[6]the halophilic bacterium screening in the soil of Deng Cong Great Salt lake can effectively be processed high salt phenolic wastewater in sbr reactor device.Yet, the most of focus of the report of high-salt wastewater are the removal of organism and nitrogen both at home and abroad, at present almost not to having the report of dephosphorization ability bacterial classification under high salt condition, the synchronous denitrification dephosphorizing of realizing waste water under high salinity condition is a challenging problem especially.
The present invention isolates a strain salt vibrios (Salinivibrio), finds that it has the ability that resistance to high salt has heterotrophic nitrification-aerobic denitrification concurrently; Further find that this bacterium has the ability of synchronous dephosphorization concurrently under single aerobic condition.Utilize this class to there is physiological property and the metabolic mechanism of the bacterium of special property, it based on nitrifying process, can be the physiological behavior of heterotrophic organism, and denitrification and dephosphorization process can be carried out under aerobic condition, make under the same aerobic environment of high salt condition, to complete denitrogenation dephosphorizing, can overcome preferably the contradictory problems existing in the above-mentioned traditional biological processing of mentioning, complete thus the present invention.
Summary of the invention
The object of the present invention is to provide a kind of high salt to have the bacterial strain of heterotrophic nitrification-aerobic denitrification and aerobic phosphate uptake ability and the application in wastewater treatment thereof concurrently.
Salt vibrios provided by the invention (Salinivibrio) bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (being called for short CGMCC) on March 29th, 2012, and preserving number is CGMCC No.5946.
Bacterial strain provided by the present invention, has following phenotypic characteristic: at 25-35 ℃, cultivate after 16-32h bacterium colony smooth surface, non-pigment on nutrient agar; By being negative under the microscope after gramstaining, it is shaft-like that thalline is, straight or bend to arcuation, and bacterium colony is less, flat, translucent.
The 16S rRNA gene sequence characteristic of this bacterial strain: its 16S rRNA has the nucleotide sequence as shown in sequence in sequence table 1, sequence length is 1409bp.
According to its morphological specificity and physiological and biochemical property and the result for retrieval of 16S rRNA gene order in Genbank thereof, identify that this bacterial strain is salt vibrios (Salinivibrio).According to this bacterial strain salt resistant character experimental result, salt vibrios (Salinivibrio) salt tolerant scope (take NaCl) is 1-13%.
Salt vibrios provided by the present invention (Salinivibrio) can, under high salt condition, be take organism as electron acceptor(EA), NH
4 +for electron donor, by NH
4 +be oxidized to NO
2 -or NO
3 -; Can, under aerobic condition, take organism as electron donor, NO
2 -or NO
3 -for electron acceptor(EA), be reduced to nitrogen; Can also be under aerobic condition, inorganic phosphorus is taken in and in body, is converted into self component and then realizes the object of removing phosphor in sewage element.
Salt vibrios provided by the present invention (Salinivibrio) can be used for synchronous denitrification dephosphorizing, in actual applications, bacterial strain can be placed in to high-salt wastewater and realize the synchronous object of removing of nitrogen phosphorus.
The carbon-nitrogen ratio of described waste water can be 3.7-12, is preferably 9-12.
The temperature of described waste water can be 20-40 ℃, is preferably 25-30 ℃.
The pH of described waste water can be 6.5-8.0, is preferably 6.5-7.5.
Salt vibrios of the present invention (Salinivibrio) and application thereof have following beneficial effect compared with prior art:
(1) salt vibrios of the present invention (Salinivibrio) bacterial strain is strong to the tolerance of high salt, can under high salt, aerobic condition, realize the synchronous removal of nitrogen phosphorus under aerobic condition, solve biological carbon and phosphorous removal in the restriction of high salt pair traditional biological treating processes and traditional wastewater treatment and need to take anaerobic phosphorus release, anoxic denitrification, aerobic nitrification to inhale the bottleneck problem of phosphorus staging treating;
(2) nitrification and denitrification coupling is carried out, and the basicity producing in denitrification process can well make up the acidity producing in nitrifying process, and whole process regulates pH without adding alkali; Compare Autotrophic nitrification bacterium, the growth velocity of allotrophic nitrobacteria is fast, cell yield is high, can effectively solve the increment of Autotrophic nitrification bacterium slowly, the long problem of system hydraulic detention time;
(3) denitrogenation and dephosphorization are synchronously carried out, and have solved denitrifying bacteria and the polyP bacteria race problem to carbon source;
(4) adopt the present invention, can complete the synchronous removal of carbon nitrogen phosphorus, need not build new reactor, simplified to greatest extent technical process, saved the cost of equipment and investment, therefore, there is good economic benefit and environmental benefit;
(5) the present invention is applicable to the denitrogenation dephosphorizing processing of high slat-containing wastewater, has a extensive future, and has good social benefit.
Below in conjunction with embodiment, describe the present invention.Embodiment is only for illustrating, and scope of the present invention is not limited with specific implementation method, but is limited by the scope of claim.
Accompanying drawing explanation
Accompanying drawing 1 salt vibrios (Salinivibrio) when salinity is 3% to nitrogen and phosphatic degradation curve
Accompanying drawing 2 salt vibrios (Salinivibrio) degradation curve to ammonia nitrogen under different salinity
Accompanying drawing 3 salt vibrios (Salinivibrio) under different salinity to phosphatic degradation curve
The nitrogen desorption curve of accompanying drawing 4 salt vibrios (Salinivibrio) under condition of different pH
The dephosphorization curve of accompanying drawing 5 salt vibrios (Salinivibrio) under condition of different pH
The nitrogen desorption curve of accompanying drawing 6 salt vibrios (Salinivibrio) under condition of different temperatures
The dephosphorization curve of accompanying drawing 7 salt vibrios (Salinivibrio) under condition of different temperatures
The nitrogen desorption curve of accompanying drawing 8 salt vibrios (Salinivibrio) under different C/N conditions
The dephosphorization curve of accompanying drawing 9 salt vibrios (Salinivibrio) under different C/N conditions
Embodiment
Below in conjunction with specific embodiment, the invention will be further described, but the present invention is not limited to following examples.
In following embodiment, if no special instructions, be ordinary method.
In following embodiment, described percentage composition if no special instructions, is quality percentage composition.
In embodiment, the method for monitoring and analyzing of various pollutents is with reference to < < water and effluent monitoring analytical procedure > > (Si Ban， China Environmental Science Press, 2002).Temperature and dissolved oxygen are measured by portable dissolved-oxygen content analyser (YSI550A, USA).Sludge concentration (MLSS) and volatile suspended solid concentration (MLVSS) are according to gravimetric determination.
The various units that use in embodiment, the unified national standard that adopts.
Embodiment 1: the synchronous denitrification dephosphorizing ability of salt vibrios (Salinivibrio) when salinity is 3% measured
To be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (being called for short CGMCC), preserving number is that salt vibrios (Salinivibrio) bacterial strain (lower same) of CGMCC No.5946 is inoculated in the LB substratum of 1L, prevent the intrusion of miscellaneous bacteria and keep the growth vigor of thalline, carry out enrichment culture.The bacterium liquid that cultivation is obtained is centrifugal, with sterilized water washing three times, makes optical density(OD) (OD
600) be the bacteria suspension of 1-2.
Getting the above-mentioned bacteria suspension of 10mL adds the test media of two salinity that contain 90mL (take NaCl) as 3% (every liter containing 0.94g glucose, 0.153g NH
4cl, 0.035g KH
2pO
4, 0.1gMgSO
47H
2o, 0.006g FeSO
47H
2o, pH7.0 ~ 7.5) in Erlenmeyer flask, by 9 layers of gauzes sealing, shaking culture in the shaking table of 30 ℃ of 150rpm.The substratum of not inoculating bacteria suspension carries out experiment under equal conditions as blank.In 0h, 6h, 12h, 18h and the 24h reaction solution that takes a morsel, wherein a part is directly used in and measures thalline optical density(OD), and rest part is centrifugal 5min under 8000rpm, gets supernatant liquor and measures various nitrogenous compounds and phosphatic concentration.
Experimental result is shown in accompanying drawing 1.Salt vibrios (Salinivibrio) bacterial strain well-grown under high salt condition, and ammonia nitrogen and phosphoric acid salt are had to stronger degradation capability.In 24h, bacterial strain is in logarithmic phase, and meanwhile ammonia nitrogen and phosphatic degraded synchronously carried out, and during 24h, clearance reaches maximum value 93.6 and 100%.In addition, in whole degradation process, the concentration of nitre nitrogen and nitrite nitrogen is without obvious accumulation, and in waste water, most ammonia nitrogen is converted into gaseous product by heterotrophic nitrification-aerobic denitrification effect.In addition, in 24h, phosphatic concentration drops to 0mg/L by initial 10mg/L, illustrates when this bacterial strain has peculiar nitrated-aerobic denitrification ability under high salt condition and has stronger phosphorus removal functional concurrently.
Embodiment 2: the Nitrogen/Phosphorus Removal of bacterial strain under different salinity condition
Take glucose as carbon source, and ammonia nitrogen is nitrogenous source, and the vibrios of salt described in embodiment (Salinivibrio) bacterial strain is measured the removal ability of ammonia nitrogen under different salinity.Concrete implementation step is as follows:
Salt vibrios (Salinivibrio) inoculation is respectively to 1%, 3%, 5%, 8%, 10% and 13% 100ml minimal medium (0.94g glucose, 0.153g NH in salinity (in NaCl)
4cl, 0.035g KH
2pO
4, 0.1g MgSO
47H
2o, 0.006g FeSO
47H
2o, 30g-150g NaCl, pH7.0 ~ 7.5) in, in the shaking table of 30 ℃ of 150rpm, carry out preculture.Until strain growth, to logarithmic phase during the later stage, get 10ml bacterium liquid and access fresh salinity (in NaCl) and be respectively in 1%, 3%, 5%, 8%, 10% and 13% 90ml minimal medium, in the shaking table of 30 ℃ of 150rpm, carry out shaking culture.The substratum of not inoculating bacteria suspension carries out experiment under equal conditions as blank.In 6h, 12h, 18h, 24h, 36h and the 48h reaction solution that takes a morsel, wherein a part is directly used in and measures thalline optical density(OD), and rest part is centrifugal 10min under 8000rpm, gets supernatant liquor and measures nitrogenous compound and phosphatic concentration.
As shown in Figure 2, initial nitrogen concentration is 40mg/L, and when salinity is respectively 1%, 3%, 5%, 8%, 10% and 13%, after 48h, the concentration of ammonia nitrogen is respectively 31.08mg/L, 3.56mg/L, 4.53mg/L, 6.78mg/L, 7.23mg/L and 36.72mg/L; When salinity is 1%, salt vibrios (Salinivibrio) bacterial strain is not very desirable to the removal effect of ammonia nitrogen, and 48h clearance is only 22.03%; When salinity is 3%, salt vibrios (Salinivibrio) bacterial strain is best to the removal effect of ammonia nitrogen, and the clearance of its 24h and 48h reaches respectively 93.59% and 91.10%; After this along with salinity raises, bacterial strain is on a declining curve to the removal of ammonia nitrogen, but until salinity is increased to 10%, it remains on more than 80% to the clearance of ammonia nitrogen always; When salinity reaches 13%, bacterial strain obviously declines to the removal effect of ammonia nitrogen, and clearance is only 8.20%, infers that reason may be that under this salinity, this strain growth speed declines, and produces stronger restraining effect to its degradation capability.Hence one can see that, and the optimum salinity of salt vibrios (Salinivibrio) degradation of ammonia nitrogen is 3%, and in the scope that is 3%-10% in salinity, ammonia nitrogen is all had to good removal effect.
As shown in Figure 3, this bacterial strain, when salinity (in NaCl) is respectively 3%, 5%, 8%, 10%, all can effectively be removed phosphoric acid salt to result; When salinity is 1% and 13%, the phosphor-removing effect of bacterial strain is undesirable.Wherein the phosphorus removal property when salinity is 3% is best, removes speed also the fastest, and phosphatic maximum material removal rate reaches 93.6%.As can be seen here, this bacterial strain is for there being the Facultative Halophiles of well tolerable ability to high salt, and it can effectively realize biological phosphate-eliminating under high salt condition.
The denitrogenation dephosphorizing experiment of embodiment 3:. bacterial strain under condition of different pH
Salt vibrios (Salinivibrio) inoculation in 3% minimal medium, is carried out to preculture in the salinity of 100ml (take NaCl) in the shaking table of 30 ℃ of 150rpm.Until strain growth to logarithmic phase during the later stage, getting 10ml bacterium liquid accesses in fresh 90ml minimal medium (with embodiment 2), regulate medium pH to be respectively in the scope of 6.5-7.0,7.0-7.5 and 7.5-8.0, in the shaking table of 30 ℃ of 150rpm, carry out shaking culture.The substratum of not inoculating bacteria suspension carries out experiment under equal conditions as blank.In 6h, 12h, 18h, 24h, 36h and the 48h reaction solution that takes a morsel, wherein a part is directly used in and measures thalline optical density(OD), and rest part is centrifugal 10min under 8000rpm, gets supernatant liquor and measures various nitrogenous compounds and phosphatic concentration.
From Fig. 4, Fig. 5, salt vibrios (Salinivibrio) bacterial strain is 6.5-7.0 at pH, and denitrogenation dephosphorizing performance is best, and 24h reaches respectively 93.6 and 100% to ammonia nitrogen and phosphatic clearance.But along with the rising of pH, bacterial strain denitrogenation dephosphorizing rate reduces.When pH is within the scope of 7.5-8.0, bacterial strain is not very desirable to ammonia nitrogen and phosphatic removal effect.Hence one can see that, and the suitableeest pH scope of this bacterial strain Nitrogen/Phosphorus Removal is 6.5-7.0, and the rising meeting of pH value produces restraining effect to its Nitrogen/Phosphorus Removal.
Embodiment 4: the denitrogenation dephosphorizing experiment of bacterial strain under condition of different temperatures
Salt vibrios (Salinivibrio) inoculation in 3% minimal medium (with embodiment 2), is carried out to preculture in the salinity of 100ml (take NaCl) in the shaking table of 30 ℃ of 150rpm.Until strain growth, to logarithmic phase, during the later stage, get 10ml bacterium liquid and access in fresh 100ml minimal medium, regulate shaking table culture temperature to be respectively 20 ℃, 30 ℃ and 40 ℃, 150rpm shaking culture.The substratum of not inoculating bacteria suspension carries out experiment under equal conditions as blank.In 6h, 12h, 18h, 24h, 36h and the 48h reaction solution that takes a morsel, wherein a part is directly used in and measures thalline optical density(OD), and rest part is centrifugal 10min under 8000rpm, gets supernatant liquor and measures various nitrogenous compounds and phosphatic concentration.
From Fig. 6, Fig. 7, when temperature is respectively 20 ℃, 30 ℃ and 40 ℃, after 48h, the concentration of ammonia nitrogen is respectively 6.51mg/L, 3.56mg/L and 7.40mg/L, and phosphatic concentration is respectively 2.27mg/L, 2.30mg/L and 2.83mg/L.When temperature is 30 ℃, bacterial strain is the highest to ammonia nitrogen and phosphatic clearance; The accumulation of nitrate-free nitrogen in 48h, only has micro-nitrite nitrogen to occur, its value is all no more than 0.04mg/L.Illustrate that salt vibrios (Salinivibrio) bacterial strain has wider subject range to temperature.
Embodiment 5: the denitrogenation dephosphorizing experiment of bacterial strain under different C/N conditions
Salt vibrios (Salinivibrio) inoculation in 3% minimal medium (with embodiment 2), is carried out to preculture in the salinity of 100ml (take NaCl) in the shaking table of 30 ℃ of 150rpm.Until strain growth, to logarithmic phase, during the later stage, get 10ml bacterium liquid and access in fresh 100ml minimal medium, regulate C/N to be respectively 3.7,7.5 and 9.0, in the shaking table of 30 ℃ of 150rpm, carry out shaking culture.The substratum of not inoculating bacteria suspension carries out experiment under equal conditions as blank.In 6h, 12h, 18h, 24h, 36h and the 48h reaction solution that takes a morsel, wherein a part is directly used in and measures thalline optical density(OD), and rest part is centrifugal 10min under 8000rpm, gets supernatant liquor and measures various nitrogenous compounds and phosphatic concentration.
From Fig. 8, Fig. 9, when C/N is respectively 3.7,7.5 and 9, after 48h, the concentration of ammonia nitrogen is respectively 9.09mg/L, 6.94mg/L and 4.56mg/L, and phosphatic concentration is respectively 4.04mg/L, 3.16mg/L and 2.30mg/L; When C/N is 9, salt vibrios (Salinivibrio) is 3.7 and the clearance of 9 o'clock apparently higher than it at C/N to ammonia nitrogen and phosphatic clearance.
Embodiment 6: the determining of the best dephosphorization condition of bacterial strain
Adopt the pattern of shake flat experiment, according to principle of orthogonal experiment, take pH value, shaking table culture temperature, inoculum size and four factors of carbon-nitrogen ratio is factor of influence, and 4 factor 3 levels of having set up amount to 9 orthogonal tests to determine the best dephosphorization condition of bacterial strain salt vibrios (Salinivibrio).The substratum of selecting in experiment is except glucose and ammonium chloride content change as required, and all the other components and concentration are identical with embodiment 1 without exception.
Orthogonal and the results are shown in Table shown in 1.As shown in Table 1, the power that affects of four factor pair salt vibrios (Salinivibrio) phosphor-removing effect is: pH> carbon-nitrogen ratio > temperature > inoculum size; The best phosphor-removing effect of salt vibrios (Salinivibrio) is combined as A
2b
2c
3d
2, corresponding optimal conditions is that pH is 6.5-7.0, and temperature is 30 ℃, and inoculum size is 10%, and carbon-nitrogen ratio is 9.
Table 1 orthogonal and result
Claims (1)
1. a high salt has the application of salt vibrios (Salinivibrio sp.) bacterial strain in wastewater treatment of heterotrophic nitrification-aerobic denitrification and phosphorus removal functional concurrently, it is characterized in that: this salt vibrios bacterial strain preserving number is CGMCC No.5946, and this bacterial strain can be realized the synchronous removal to N, P under high salt, single aerobic condition; Wherein, the salinity of described waste water, in NaCl, scope is 1-13%; The carbon-nitrogen ratio scope of described waste water is 3.7-12; The pH scope of described waste water is 6.5-8.0.
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