CN103923834A

CN103923834A - Method suitable for rapid enrichment culture of halophilic and halotolerant bacteria

Info

Publication number: CN103923834A
Application number: CN201310740826.7A
Authority: CN
Inventors: 胡蓉; 曾凡付; 杨泰山; 张清华
Original assignee: XINJIANG DELAN CO Ltd
Current assignee: XINJIANG DELAN CO Ltd
Priority date: 2013-12-27
Filing date: 2013-12-27
Publication date: 2014-07-16

Abstract

The invention provides a method suitable for the rapid enrichment culture of halophilic and halotolerant bacteria. The method comprises the following steps: 1, carrying out 10 times dilution on filtered, decolored and deoiled wastewater by using distilled water; 2, preparing a culturing medium composed of 3-6g of hydrolyzed casein, 5-10g of a beef extract, 5-10g of tryptone, 0.5-1g of KHPO4, 0.5-1g of K2HPO4, 2-5g of KCl, 0.05g of MgSO4.7H2O, 0.05g of CaCl2 and 0.05g of FeSO4.7H2O; 3, dissolving components composing the culturing medium in 1L of the diluted wastewater solution obtained in step 1, adding 10g of NaCl, and adjusting the pH value to 7.0-8.0 by using NaOH; and 4, placing a solution obtained in step 3 on a shaking bed, culturing at 125rpm at a temperature of 30DEG C, adding 11g of NaCl and 100ml of the culturing medium obtained in step 2 every 14h, and culturing for 70h to obtain a halophilic and halotolerant bacterium mixture.

Description

A kind of method that adapts to the cultivation of halophilic bacterium Facultative Halophiles fast enriching

Technical field

The present invention relates to high-salt wastewater processing, the high-salt wastewater that organic chemical industry, Coal Chemical Industry, textile printing and dyeing, oil production etc. are produced, implements the method for fast enriching cultivation and obtains halophilic bacterium Facultative Halophiles, and effect is very remarkable.

Background technology

High-salt wastewater refer to salt massfraction>=1% contain the organic waste water of finite concentration, wherein contain a large amount of inorganic salt, as Cl ^-, S0 ₄ ^2-, Na, Ca ²⁺plasma, the method of processing at present high slat-containing wastewater mainly contains Physical, physico-chemical processes and biological process, wherein the processing costs of Physical and physico-chemical processes is relatively high, therefore, biological process has become the prefered method of most of high-salt wastewater treatment process, but, inorganic salt in high salt industrial waste water have stronger restraining effect to general microorganism, different bioreactor for treatment high-salt wastewaters effect difference, in common biological process, most of microbe can only tolerate the waste water of saliferous below 3%, in the time that saltiness is greater than 5%, its treatment effect can reduce greatly, therefore, utilizing common micro-organisms to process high-salt wastewater need to dilute, this can cause the waste of water resources, so far, urgently develop without desalination, the novel process of diluting and directly carry out a biological disposal upon, utilize Facultative Halophiles or halophilic bacterium inoculation to improve the technique of aerobic activated sludge, also cultivated through screening, produce stable treatment effect.

Literature search disclose: with the cultural method of a Chinese patent CN101985601A(salt tolerant bacterial classification) compared with, in patent CN101985601A, medium salts concentration is directly adjusted to target salt concn, suddenly the salt concn pressure increasing will cause the mortality of bacterial classification, and in the present invention, progressively increase salt concn pressure, make microorganism resource in system obtain maximum reservation.Simultaneously, compared with substratum compound method in patent CN101985601A, the present invention also improves, adopt target wastewater dilution liquid to replace tap water, reach the at utmost object of the environment of simulated target waste water, therefore go out can salt tolerant in easier enrichment, can adapt to again the bacterial strain of target waste water environment.

Present method is applicable to the novel method of halophilic bacterium, Facultative Halophiles fast culture, what can obtain being fast adapted to high-salt wastewater micro-ecological environment has a liking for salt, salt resistant function bacterial classification, with this bacterium inoculation of activated-sludge, improve the treatment effect of high-salt wastewater, there is important production and application and be worth.

Summary of the invention

The object of the invention is to: the cultural method of having a liking for salt, salt resistant function bacterial classification that is adapted to different high-salt wastewater micro-ecological environments is provided, with this obtain can inoculation of activated-sludge bacterial classification, improve the treatment effect of high-salt wastewater.

The object of the present invention is achieved like this: a kind of method that adapts to the cultivation of halophilic bacterium Facultative Halophiles fast enriching, implement step by step;

Step 1 is by the waste water after filtration, decolouring, oil removing, with 10 times of distilled water dilutings, for subsequent use;

Step 2 substratum: by 3-6g caseinhydrolysate, 5-10g extractum carnis, 5-10g Tryptones, 0.5-1g KHPO ₄, 0.5-1g K ₂hPO ₄, 2-5g KCl, 0.05g MgSO ₄7H ₂o, 0.05g CaC1 ₂, 0.05g FeSO ₄7H ₂o composition;

Step 3 is got step 2) component be dissolved in 1L step 1) wastewater dilution liquid, add 10g NaCl, be adjusted to pH7.0-8.0 with NaOH, for subsequent use;

Step 4 is by step 3) be placed on shaking table, under 125rpm condition, cultivate, 30 DEG C of temperature, at interval of 14h, under aseptic condition, add 11g NaCl and step 2) substratum 100ml, and cultivate 70h, obtain halophilic bacterium Facultative Halophiles mixture.

Described method, adopts this Facultative Halophiles to process the active sludge of high-salt wastewater, and the place to go rate of COD has improved 15%, TN and TP clearance have improved 10%.

Technique effect of the present invention be can obtain at short notice maximum biomass have a liking for salt, salt tolerant bacterial classification, the scheme of the gradient rising salt concn of taking, avoid the excessive loss that causes a large amount of microorganism resource of suddenly increasing of salt concn pressure, simultaneously, adopt target wastewater dilution liquid to replace tap water, the environment of more effective simulated target waste water, easier enrichment goes out can salt tolerant, can adapt to again the bacterial strain of target waste water environment.

The inventive method is cultivated the Facultative Halophiles obtaining and is linked in the active sludge of processing target high-salt wastewater, and the place to go rate of COD has improved 15%, TN and TP clearance have improved 10%, for the biological treatment of different types of high-salt wastewater provides basis, shows technical progress.

Embodiment

Below in conjunction with embodiment, the present invention will be described in detail.

Embodiment 1

Get the gorgeous waste water that fiber company limited workshop produces that reaches, its saltiness is 3.5%, and this water is filtered, adopted after 0.8mm granulated apricot shell activated carbon decolorizing, de-oiling with 100 object gauze, with 10 times of distilled water dilutings, for subsequent use;

1) get caseinhydrolysate 5g, extractum carnis 8g, Tryptones 5g, KHPO ₄0.5g, K ₂hPO ₄0.5g, KCl2g, MgSO ₄7H ₂o0.05g, CaC1 ₂0.05g, FeSO ₄7H ₂o0.05g, is mixed to get substratum, then this substratum is placed in to 1L wastewater dilution liquid, adds NaCl10g, with NaOH adjustment pH to 7.0;

2) step 1) solution is placed on shaking table, shaking table is under 130rpm condition, and 25 DEG C of culture temperature every 15h, are added substratum 100ml and 12g NaCl in step 1) under aseptic condition, cultivates 75h.

3) this system influent COD is 1500mg/L, after ordinary method is processed, COD clearance is 73.3%, cultivating with this cultural method the halophilic bacterium, the Facultative Halophiles that obtain is linked in the gorgeous active sludge that reaches fiber company limited pulp workshop institute high-salt wastewater of processing, the clearance of COD has improved 15%, TN and TP clearance have improved 10%

Embodiment 2

Get the waste water that certain black petrochemical industry produces, its saltiness is 2%, by after these water 100 object gauze filtrations, 1mm granulated apricot shell activated carbon decolorizing, de-oiling, with 10 times of distilled water dilutings, for subsequent use;

1) get caseinhydrolysate 3g, extractum carnis 10g, Tryptones 5g, KHPO ₄0.6g, K ₂hPO ₄0.6g, KCl3g, MgSO ₄7H ₂o0.05g, CaC1 ₂0.05g, FeSO ₄7H ₂o0.05g is mixed to get substratum, then substratum is placed in to 1L wastewater dilution liquid, adds NaCl10g, with NaOH adjustment pH to 7.0;

2) solution of step 1) is placed on shaking table, shaking table is under 120rpm condition, and 37 DEG C of culture temperature every 12h, are added substratum 100ml and 10g NaCl in step 1) under aseptic condition, cultivates after 60h.

3) to produce high-salt wastewater COD be 1400mg/L to black petrochemical industry, after using ordinary method to process, clearance is 60%, adopt halophilic bacterium, Facultative Halophiles that this cultural method obtains to be linked in the active sludge of processing high-salt wastewater that black petrochemical industry produces, the place to go rate of COD has improved 15.5%, TN and TP clearance have improved 11%.

The caseinhydrolysate that the inventive method is selected is purchased from Beijing Hua Maike biotechnology limited liability company; Extractum carnis and Tryptones are all purchased from Beijing extensive and profound in meaning star biotechnology limited liability company; KHPO ₄and K ₂hPO ₄purchased from Hedong District, Tianjin red rock chemical reagent work; MgSO ₄7H ₂o and FeSO ₄7H ₂o is purchased from Tianjin Bo Di Chemical Co., Ltd..

Claims

1. adapt to the method that halophilic bacterium Facultative Halophiles fast enriching is cultivated, it is characterized in that: implement step by step;

Step 2 substratum: by 3-6 g caseinhydrolysate, 5-10g extractum carnis, 5-10g Tryptones, 0.5-1g KHPO ₄, 0.5-1 g K ₂hPO ₄, 2-5 g KCl, 0.05g MgSO ₄7H ₂o, 0.05g CaC1 ₂, 0.05g FeSO ₄7H ₂o composition;

Step 3 is got step 2) component be dissolved in 1L step 1) wastewater dilution liquid, add 10g NaCl, be adjusted to pH 7.0-8.0 with NaOH, for subsequent use;

Step 4 is by step 3) be placed on shaking table, under 125 rpm conditions, cultivate, 30 DEG C of temperature, at interval of 14h, under aseptic condition, add 11g NaCl and step 2) substratum 100ml, and cultivate 70h, obtain halophilic bacterium Facultative Halophiles mixture.

2. method according to claim 1, is characterized in that: adopt this Facultative Halophiles to process the active sludge of high-salt wastewater, the place to go rate of COD has improved 15%, TN and TP clearance have improved 10%.

3. method according to claim 1, is characterized in that: the preparation of selecting is commercially available prod.