CN105995375A - Method for effectively killing viable but nonculturable state microorganisms generated in ultrasonic sterilization process - Google Patents
Method for effectively killing viable but nonculturable state microorganisms generated in ultrasonic sterilization process Download PDFInfo
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- CN105995375A CN105995375A CN201610365148.4A CN201610365148A CN105995375A CN 105995375 A CN105995375 A CN 105995375A CN 201610365148 A CN201610365148 A CN 201610365148A CN 105995375 A CN105995375 A CN 105995375A
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- ultrasonic
- electric potential
- ultrasonic sterilization
- potential water
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/005—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by heating using irradiation or electric treatment
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/358—Inorganic compounds
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- Life Sciences & Earth Sciences (AREA)
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- Food Science & Technology (AREA)
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- Inorganic Chemistry (AREA)
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Abstract
The invention discloses a method for effectively killing viable but nonculturable state microorganisms generated in ultrasonic sterilization process. The method comprises the following steps: staphylococcus aureus is washed with a stroke-physiological saline solution and then is swirled into slightly acid electrolyzed water; and the bacterial suspension is subjected to ultrasonic treatment immediately. According to the method provided by the invention, staphylococcus aureus bacteria are swirled into the slightly acid electrolyzed water, and then the ultrasonic treatment is carried out, so that the proportion of viable but nonculturable state bacteria is greatly reduced. The experiment of the invention proves that the technology of combining ultrasonic wave with the slightly acid electrolyzed water is used for treating bacteria, can effectively reduce the proportion of viable but nonculturable state bacteria in the ultrasonic sterilization process, enhances the safety of the ultrasonic sterilization process and is beneficial for promoting the industrial application of ultrasonic sterilization technology.
Description
Technical field
The present invention relates to a kind of work effectively killed and produce during ultrasonic sterilization can not cultivation conditions
The method of microorganism, advantageously reduce alive can not the ratio of cultivation conditions antibacterial, optimize ultrasonic sterilization
Technique, the industry application of further genralrlization ultrasonic sterilization technology, belong to Food Engineering Development field.
Background technology
As emerging non-thermal sterilization technology, ultrasonic sterilization has the features such as energy consumption is low, the time is short, with
Other non-thermal sterilization technologies are compared, and ultrasound wave, during killing microorganisms, can effectively reduce sub-lethal
The potential threat that cell causes, is widely used in fields such as waste water process, drinking water disinfections at present.
Ultrasound wave for food sterilization in addition to effectively preserving food nutrient composition and natural color,
Food can also be produced such as homogenizing, urge the multiple effects such as old, cracking macromolecular substances, it is thus possible to
Preferably improve and improve food quality.But, research shows that supersound process can be formed by Induction of bacterial
That lives can not cultivation conditions.Live can not the antibacterial of cultivation conditions cannot be normal in conventional culture medium
Growth and breeding, it is possible to escape conventional sense and cause false-negative result, but still there is vital sign
And metabolic activity, under appropriate conditions can recovery regrowth, regain infection ability, become danger
" the recessive pollution source " of evil food safety, makes ultrasonic sterilization technology at the application surface of food processing field
Face challenge.
Effectively kill in ultrasonication produce work can not cultivation conditions antibacterial, be improve ultrasonic
Bactericidal effect, strengthens the important means of the safety of supersound process food, but at present about ultrasound wave
Application in food sterilization is confined to assess the antibacterial that ultrasound wave is killed completely, and is not directed to ultrasonic
The work produced in processing procedure can not the antibacterial of cultivation conditions.The present invention uses ultrasonic combined weak acid
Property electric potential water technology effectively kill alive can not the antibacterial of cultivation conditions, live not after reducing supersound process
Can the probability of cultivation conditions antibacterial recovery regrowth under optimum conditions.
Summary of the invention
It is an object of the invention to provide and a kind of effectively kill the work produced during ultrasonic sterilization not
Can the method for cultivation conditions microorganism, live after reducing supersound process can not cultivation conditions antibacterial suitably
Under the conditions of the probability of recovery regrowth.
A kind of effectively kill the work that produces during ultrasonic sterilization can not the side of cultivation conditions microorganism
Method, including: after being washed with physiological saline solution by staphylococcus aureus, vortex is to faintly acid electric potential water
In, at once this bacterial suspension is carried out supersound process.
The method that the present invention provides, is to faintly acid electric potential water by staphylococcus aureus thalline vortex
In, carry out supersound process the most again, greatly reduce alive can not the ratio of cultivation conditions antibacterial.This
Invention it is demonstrated experimentally that utilize ultrasonic combined faintly acid electric potential water technical finesse antibacterial, can effectively drop
Live during low ultrasonic sterilization can not the ratio of cultivation conditions antibacterial, enhance ultrasonic sterilization technique
Safety, is conducive to promoting the industry application of ultrasonic sterilization technology.
Preferably, the mass fraction of described physiological saline solution is 0.8～0.9%.More preferably
0.85%.
Preferably, the effective chlorine density of described faintly acid electric potential water is 1～3mg/L.More preferably
2mg/L。
Preferably, the pH of described faintly acid electric potential water is 6～6.5, and ORP is 850～880mV；Enter
One step preferably, the pH=6.1, ORP=867.4mV of described faintly acid electric potential water.
It is further preferred that described faintly acid electric potential water is by circulating concentration faintly acid electric potential water maker
Electrolysis 0.1%HCl and 0.03%NaCl solution 1min is prepared.
Compared with tradition electric potential water maker, circulation concentrates faintly acid electric potential water maker by adding one
The electric potential water of generation is pressed in electrolysis bath and carries out repeating electrolysis and obtain circulating dense by individual circulating device again
Contracting faintly acid electric potential water.By electrolysis 0.1%HCl and 0.03%NaCl solution 1min, it is prepared into
To faintly acid electric potential water (pH=6.1, ORP=867.4mV, ACC=30mg/L).PH-ORP surveys
Determining instrument and be used for measuring pH and the ORP value of faintly acid electric potential water, effective chlorine analyzer measures faintly acid electricity
Position water effective chlorine density (ACC), obtains required faintly acid electric potential water by distilled water gradient dilution
Concentration.
Conventional ultrasonic sterilization be bacterial suspension in 0.85% normal saline, then bacteria suspension is carried out
Supersound process, present invention faintly acid electric potential water is replaced normal saline, is utilized its strong oxidizing property can not only
Enough improve germicidal efficiency, moreover it is possible to during effective reduction ultrasonic sterilization, the work of generation can not cultivation conditions
Antibacterial.
Preferably, the condition of described supersound process is: processing frequency is 15～25kHz, processes power
Being 300～500W, the process time is 1-10min, treatment temperature 15～25 DEG C.
It is further preferred that the condition of described supersound process is: process frequency is 20kHz, process merit
Rate is 400W, and the process time is 1min, and treatment temperature is 20 DEG C.
Or: process frequency is 20kHz, and process power is 400W, and the process time is 10min, place
Reason temperature is 20 DEG C.
Or: process frequency is 20kHz, and process power is 400W, and the process time is 5min, place
Reason temperature 20 DEG C.
This preferred ultrasound condition and faintly acid electric potential water have more preferable synergy between suspending, optimum
Selection of land is combined as:
The pH=6.1 of described faintly acid electric potential water, ORP=867.4mV, ACC=2mg/L；Described super
The condition of sonication is: process frequency is 20kHz, and process power is 400W, and the process time is 1
Min, treatment temperature is 20 DEG C.The work of staphylococcus aureus can not can be cultivated shape under the conditions of Gai
State bacterium ratio is reduced to about 5% from about 30%.
After ultrasonic in combination faintly acid electric potential water processes, further comprising the steps of: detection faintly acid current potential
Water add before and after staphylococcus aureus viable count and can breeding bacteria；Wherein, detection golden yellow
Staphylococcus viable count uses the double dye method of PI/cFDA, and staphylococcus aureus can be adopted by breeding bacteria in detection
Use colony counting method.
The experiment proves that, the present invention develops during one effectively kills ultrasonic sterilization and produces
Work can not the method for cultivation conditions microorganism, live after reducing supersound process can not cultivation conditions thin
The probability of bacterium recovery regrowth under optimum conditions, optimizes ultrasonic sterilization technique, favorably further
In the industry application promoting ultrasonic sterilization technology.
Detailed description of the invention
Below in conjunction with specific embodiment, further illustrate the present invention.
Embodiment 1
One ultrasonic in combination faintly acid electric potential water processes
(1) prepared by bacterial suspension
Staphylococcus aureus reference culture ATCC 25923 is inoculated in broth medium,
37 DEG C of shaking tables are cultivated to exponential phase.
Experimental group: the bacterium of the physiological saline solution washing exponential phase using mass fraction to be 0.85%
Body twice, is then resuspended to thalline in the faintly acid electric potential water of 2mg/mL, makes cell concentration be about
108CFU/mL, as experimental group bacterial suspension；
Matched group: the bacterium of the physiological saline solution washing exponential phase using mass fraction to be 0.85%
Body twice, is then resuspended to thalline in the physiological saline solution of 0.85%, makes cell concentration be about 108
CFU/mL, as a control group bacterial suspension；
(2) supersound process
Immediately staphylococcus aureus suspension is entered with ultrasound wave SCIENTZ-IID type sterilizing unit
Row processes, concrete steps: loaded by 30mL bacterial suspension in glass ultraphonic pipe, a diameter of 10mm
Ultrasonic probe go deep into liquid level 2cm, ultraphonic pipe is put in 20 DEG C of water-baths, bacterium solution is surpassed
Sonication, process frequency is 20kHz, and process power is 100W, and the process time is 1min,
Rear use nertralizer (0.5% sodium thiosulfate and 0.85% sodium chloride solution) stops sterilization, obtains everywhere
Bacterium solution after reason.
Two detections
(1) viable count assay method
The double dye method of employing PI/cFDA: concrete steps: experimental group and the staphylococcus aureus of matched group
The PI of bacterium solution and 30mmol/L hatches 10min in ice bath, then with the cFDA of 50mmol/L
Hatch 10min at 37 DEG C, 4 DEG C, 6000 × g be centrifuged 10min, with the sterile physiological salt of 0.85%
The dyestuff of excess is removed in water washing, is analyzed with flow cytometer after having hatched, and each sample is received
Collect 20000 Bacteria Detection.
(2) can breeding bacteria assay method
Employing colony counting method: concrete steps: with reference to GB 4789.2-2010 method, by experimental group
Carry out gradient dilution with the staphylococcus aureus bacterium solution of matched group with 0.85% physiological saline solution, take
Diluted sample 1mL, in sterilizing plate, is poured about 20mL PCA culture medium into and is shaken up, and treats that culture medium is coagulated
It is inverted in 37 DEG C of incubators cultivation 24h after Gu, then records clump count.
(3) that lives can not cultivation conditions antibacterial
Adopt distinguish with the aforedescribed process determination experiment group and matched group viable count and can breeding bacteria, from
And calculate not cultivation conditions bacterium number alive (viable count can breeding bacteria).
Result is as follows: experimental group staphylococcus aureus viable bacteria ratio is 5.69%, can cultivate bacterium ratio
Being 0.52%, live can not cultivation conditions bacterium ratio be 5.17%.Matched group staphylococcus aureus lives
Bacterium ratio is 94.32%, and can cultivate bacterium ratio is 63.10%, and live can not cultivation conditions bacterium ratio be
31.22%.As can be seen here, the work produced during the method for the present invention can effectively kill ultrasonic sterilization
Can not cultivation conditions microorganism, reduce alive can not the ratio of cultivation conditions antibacterial, enhance ultrasonic
The safety of process for sterilizing.
Embodiment 2
One ultrasonic in combination faintly acid electric potential water processes
(1) prepared by bacterial suspension
Identical with the method for the step of in embodiment 1 (1).
(2) supersonic induced
Essentially identical with the method for the step of in embodiment 1 (2), only the process time is become
10min。
Two detections
Identical with the method for two in embodiment 1.
Result is as follows: experimental group staphylococcus aureus viable bacteria ratio is 0.09%, can cultivate bacterium ratio
Being 0.021%, live can not cultivation conditions bacterium ratio be 0.069%.Matched group staphylococcus aureus
Viable bacteria ratio is 89.40%, and can cultivate bacterium ratio is 43.65%, and that lives can not cultivation conditions bacterium ratio
It is 45.75%.As can be seen here, the method for the present invention has killed the biggest ratio when Combined Treatment 10min
Example live can not cultivation conditions antibacterial, make to live can not the ratio of staphylococcus aureus of cultivation conditions
It is down to 0.069%.
The present embodiment compared with embodiment 1, kill alive need not cultivation conditions antibacterial effect more
Good.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention are not by upper
Stating the restriction of embodiment, that is made under other any spirit without departing from the present invention and principle changes
Become, modify, substitute, combine, simplify, all should be the substitute mode of equivalence, be included in the present invention
Protection domain within.
Claims (8)
1. one kind effectively kill the work that produces during ultrasonic sterilization can not cultivation conditions microorganism
Method, it is characterised in that including: vortex after staphylococcus aureus is washed with physiological saline solution
In faintly acid electric potential water, obtain suspension, at once suspension is carried out supersound process.
Method the most according to claim 1, it is characterised in that the matter of described physiological saline solution
Amount mark is 0.8～0.9%.
Method the most according to claim 1, it is characterised in that having of described faintly acid electric potential water
Effect cl concn is 1～3mg/L.
Method the most according to claim 1, it is characterised in that the pH of described faintly acid electric potential water
Being 6～6.5, ORP is 850～880mV.
Method the most according to claim 1, it is characterised in that the condition of described supersound process is:
Processing frequency is 15～25kHz, processing power is 300～500W, and the process time is 1-10min,
Treatment temperature 15～25 DEG C.
Method the most according to claim 1, it is characterised in that the condition of described supersound process is:
Process frequency is 20kHz, and process power is 400W, and the process time is 1min, and treatment temperature is
20℃。
Method the most according to claim 1, it is characterised in that the condition of described supersound process is:
Process frequency is 20kHz, and process power is 400W, and the process time is 10min, and treatment temperature is
20℃。
Method the most according to claim 1, it is characterised in that the condition of described supersound process is:
Process frequency is 20kHz, and process power is 400W, and the process time is 5min, treatment temperature 20 DEG C.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610365148.4A CN105995375A (en) | 2016-05-27 | 2016-05-27 | Method for effectively killing viable but nonculturable state microorganisms generated in ultrasonic sterilization process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201610365148.4A CN105995375A (en) | 2016-05-27 | 2016-05-27 | Method for effectively killing viable but nonculturable state microorganisms generated in ultrasonic sterilization process |
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| CN105995375A true CN105995375A (en) | 2016-10-12 |
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| CN201610365148.4A Pending CN105995375A (en) | 2016-05-27 | 2016-05-27 | Method for effectively killing viable but nonculturable state microorganisms generated in ultrasonic sterilization process |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108522632A (en) * | 2018-06-20 | 2018-09-14 | 宁波市农业科学研究院 | A kind of defreezing method of aquatic product |
| CN112352901A (en) * | 2020-09-16 | 2021-02-12 | 浙江大学 | Application of synergy of mannosylerythritol lipids and ultrasound in inhibition of growth of drug-resistant bacteria |
| CN115094017A (en) * | 2022-06-30 | 2022-09-23 | 江南大学 | Method for inducing yeast to enter VBNC state |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103462528A (en) * | 2013-09-06 | 2013-12-25 | 浙江大学 | Fruit and vegetable cleaning and sterilizing device and method |
| CN203555652U (en) * | 2013-09-06 | 2014-04-23 | 浙江大学 | Fruit/vegetable cleaning and sterilizing device |
-
2016
- 2016-05-27 CN CN201610365148.4A patent/CN105995375A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103462528A (en) * | 2013-09-06 | 2013-12-25 | 浙江大学 | Fruit and vegetable cleaning and sterilizing device and method |
| CN203555652U (en) * | 2013-09-06 | 2014-04-23 | 浙江大学 | Fruit/vegetable cleaning and sterilizing device |
Non-Patent Citations (1)
| Title |
|---|
| 葛枝，等: "超声波结合弱酸性电位水处理改善商熟期草莓采后品质", 《农业工程学报》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108522632A (en) * | 2018-06-20 | 2018-09-14 | 宁波市农业科学研究院 | A kind of defreezing method of aquatic product |
| CN108522632B (en) * | 2018-06-20 | 2024-03-26 | 宁波市农业科学研究院 | Thawing method of raw water product |
| CN112352901A (en) * | 2020-09-16 | 2021-02-12 | 浙江大学 | Application of synergy of mannosylerythritol lipids and ultrasound in inhibition of growth of drug-resistant bacteria |
| CN112352901B (en) * | 2020-09-16 | 2022-06-10 | 浙江大学 | Application of synergy of mannosylerythritol lipids and ultrasound in inhibition of growth of drug-resistant bacteria |
| CN115094017A (en) * | 2022-06-30 | 2022-09-23 | 江南大学 | Method for inducing yeast to enter VBNC state |
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Application publication date: 20161012 |
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