CN106039353A - Physical-chemical combined cronobacter killing method and application - Google Patents
Physical-chemical combined cronobacter killing method and application Download PDFInfo
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- CN106039353A CN106039353A CN201610375333.1A CN201610375333A CN106039353A CN 106039353 A CN106039353 A CN 106039353A CN 201610375333 A CN201610375333 A CN 201610375333A CN 106039353 A CN106039353 A CN 106039353A
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- 230000002147 killing effect Effects 0.000 title abstract description 15
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- KFSLWBXXFJQRDL-UHFFFAOYSA-N Peracetic acid Chemical group CC(=O)OO KFSLWBXXFJQRDL-UHFFFAOYSA-N 0.000 claims abstract description 52
- OSVXSBDYLRYLIG-UHFFFAOYSA-N dioxidochlorine(.) Chemical compound O=Cl=O OSVXSBDYLRYLIG-UHFFFAOYSA-N 0.000 claims abstract description 42
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims abstract description 41
- 230000001954 sterilising effect Effects 0.000 claims abstract description 29
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 29
- 239000000645 desinfectant Substances 0.000 claims abstract description 23
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- 241000196324 Embryophyta Species 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/16—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using chemical substances
- A61L2/18—Liquid substances or solutions comprising solids or dissolved gases
- A61L2/186—Peroxide solutions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/02—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using physical phenomena
- A61L2/08—Radiation
- A61L2/10—Ultraviolet radiation
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- Health & Medical Sciences (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Apparatus For Disinfection Or Sterilisation (AREA)
Abstract
The invention discloses a physical-chemical combined cronobacter killing method and application and belongs to the technical field of sterilization methods. The physical-chemical combined cronobacter killing method comprises the steps that a disinfectant is sprayed on an objected to be sterilized and acts for 20-40 seconds, then irradiation treatment is performed by utilizing ultraviolet rays, and after the irradiation dose is up to 0.2-0.3 J/cm<2>, ultraviolet irradiation treatment is stopped and a neutralizer is sprayed on the objected to be sterilized; the disinfectant is peracetic acid, chlorine dioxide or hydrogen peroxide. The method can have a good cronobacter killing effect under the circumstances of low disinfectant usage amount and low ultraviolet irradiation dose. The production costs can be reduced, and the food safety risk can be also reduced under the situation that the safety of processed products is ensured.
Description
Technical field
The present invention relates to physical chemistry combined sterilizing method and the application of a kind of Cronobacter sakazakii, belong to method for disinfection technology
Field.
Background technology
Cronobacter sakazakii (Cronobacter) original name Enterobacter sakazakii (Enterobacter sakazakii), is baby
A class pathogenic bacterium in prescription emulsifiable powder (powdered infant formula, PIF), it is possible to cause neonate by polluting PIF
Infecting, fatality rate is up to 40%-80%.
The rugged Cronobacter sakazakii of slope is to have the foodborne pathogens of high risks, can cause serious septicemia, gangrenosum acne
Small intestine colon film inflammation, meningitis and sepsis.Additionally, under certain condition, this bacterium may also lead to growing up of hypoimmunity
People's bacteremia, local infection and osteomyelitis etc..Infect the high-risk group of this bacterium be neonate (less than 2.5kg) that body weight is the lightest and
Just baby's (in being born 28 days) of birth.
At present about the infant case of the infection rugged Cronobacter sakazakii of slope, concentrate on western countries.Just birth about a week
Baby easily infected by the rugged Cronobacter sakazakii of slope so that it is suffer from meningitis.When infected neonate show platycoria,
After the symptom such as ballet's disease and upper blepharoptosis, severe patient can be the most dead at several hours.Fatality rate is more than 40%, and remains
Balance reviver also can leave serious neurological sequelae, bears the torment of nervous system disease.
World Health Organization (WHO) (WHO) and Food and Agricultural Organization of the United Nations (FAO) have determined that Cronobacter sakazakii has six in belonging to
Planting bacterium and have toxicity, toxicity power there are differences.Up to the present, the mechanism of causing a disease of the rugged Cronobacter sakazakii of slope there is no final conclusion, very
Its mechanism of causing a disease is inquired into by many researchers the most from different perspectives.Mechanism of causing a disease is probably: outer membrane protein A adherent cell
Effect, the non-specific pathogenic effects of lipopolysaccharide, antibacterial interphase interaction etc..But this bacterium is caused a disease machine the most also not to have system to explain
The report of reason, the mechanism of causing a disease of the rugged Cronobacter sakazakii of slope still needs to be further elucidated with.
China's propagation about the rugged Cronobacter sakazakii of slope, the research of infection, mainly at aspects such as infant foods.Whole nation disease
The report that control center is issued at 2010-2011 claims, and Wuzhou, Guangxi province is Cronobacter sakazakii rugged to slope in 175 parts of food samples
Detecting, recall rate reaches 5.26%.In city of Jiangsu province in the monitoring of 891 parts of samples, detect that the probability of this bacterium is
2.6%.As can be seen here, the rugged Cronobacter sakazakii of slope is polluted in China's food degree and scope extensive thereof.
Infant Formula Enterprises is the primary pollution source of Cronobacter sakazakii and pollutes channel, and production and processing link is Cronobacter
The main pollution section of bacillus.Therefore, food processing equipment is effectively killed virus and there is important meaning to killing Cronobacter sakazakii
Justice.In food service industry, current method for disinfection the most all concentrates on single chemically or physically sterilization, guarantees food safety,
It is rarely employed combined sterilizing method.Although the research to physical-chemical combined sterilizing method has been reported at present, but all in just
Go out conceptual phase.Owing to processing the restriction in place, the method in a lot of researchs does not has operability.
Summary of the invention
For solving above-mentioned technical problem, the invention provides the physical chemistry combined sterilizing method of a kind of Cronobacter sakazakii,
The technical scheme taked is as follows:
It is an object of the invention to provide the physical chemistry combined sterilizing method of a kind of Cronobacter sakazakii, the method is to treating
Spraying disinfectant on sterilizing object, after disinfectant effect 20-40s, utilize ultraviolet to carry out radiation treatment, irradiation dose reaches
0.2-0.3J/cm2After, stop ultraviolet radiation treatment and utilize aseptic clear water to spray object subject to sterilization；Described disinfectant was
Fluoroacetic acid, chlorine dioxide or hydrogen peroxide.
The step of described method for disinfection is as follows:
1) in object subject to sterilization, spray peracetic acid, chlorine dioxide or hydrogenperoxide steam generator, act on 20-40s；
2) to step 1) pass through the object subject to sterilization disinfected to carry out irradiation dose being 0.2-0.3J/cm2Ultraviolet
Radiation treatment；
3) aseptic clear water object subject to sterilization to cleaning showers is utilized.
Preferably, step 1) described object subject to sterilization is plant produced equipment or packaging material.
It is highly preferred that the material of described object subject to sterilization is rustless steel, tinplate, aluminum-plastic composite membrane, glass or cement.
Preferably, step 1) mass fraction of described peracetic acid soln is 0.1-0.15%；Described chlorine dioxide adds
Amount is 500-1000ppm；The mass fraction of described hydrogen peroxide is 1.5-2.0%.
It is highly preferred that the specifically comprising the following steps that of described method
1) spraying mass fraction on plant produced equipment or packaging material is the hydrogen peroxide of 1.5-2.0%, effect
30s；
2) ultraviolet disinfection lamp is utilized to step 1) carry out purple through production equipment or the packaging material of hydrogen peroxide treatment
Outside line radiation treatment, treatment dosage reaches 0.2-0.3J/cm2Rear end；
3) utilizing aseptic clear water to step 2) through ultraviolet irradiation process production equipment or packaging material be carried out.
The above either method can be applied during the sterilization of food manufacturing apparatus and packaging material.
The beneficial effect that the present invention obtains:
Method provided by the present invention has sterilization thoroughly, workable feature, it is possible to effectively kill and pollute food
Cronobacter sakazakii on product process equipment and packaging material, it is ensured that the safety of converted products.Method provided by the present invention can
To obtain the effect preferably killing Cronobacter sakazakii in the case of using relatively low disinfectant and relatively low ultraviolet ray irradiation dose.
Both can reduce production cost, food safety risk can have been reduced.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention will be further described, but the present invention should not be limited by the examples.
Following example material therefor, reagent, method and instrument, without specified otherwise, be this area conventional material, examination
Agent, method and instrument, those skilled in the art all can be obtained by commercial channel.
Inventor's bacterial strain uses therefor is shown in Table 1, during being all isolatable from 2007 to 2015, Infant Formula Enterprises manufacturing enterprise
Processing environment and finished product, the altogether rugged Cronobacter sakazakii separation strain of 4 strain slope.
Bacterial strain uses therefor choose mainly according to two principles: first, choose the bacterial strain generation as test of main ST type
Table bacterial strain.Second, according to nearly 9 years sampled result of inventor, analyze find after spray drying, fluid bed outlet, conveyer belt etc.
Production link and infant formula are easily polluted by the rugged Cronobacter sakazakii of slope.Therefore choose these a few strain separation strains as experimental strain.
Cronobacter sakazakii bacterial strain and source thereof used by table 1 present invention
The activation culture method of above-mentioned strain is: takes out and is saved in-80 DEG C, makees with glycerol protectant containing the rugged Crow of slope
The cryopreservation tube of promise bacillus.Restore it room temperature, be inoculated in TSB fluid medium with the inoculum concentration of 2%, under the conditions of 37 DEG C
Cultivate 12h on shaking table to activate.The bacterium solution after activation is taken again solid in tryptose soya agar culture medium (TSA) with inoculating loop
Body culture medium three ride, cultivates 24h at 37 DEG C.Picking typical sizes is the yellow list bacterium colony of 1～1.5mm, is inoculated
In 10mL pancreas peptone soybean broth culture medium (TSB), carry out pure culture, continuous passage 2 times, follow-up test can be carried out.
The stand-by bacterium solution vortice of the above-mentioned 4 rugged Cronobacter sakazakii of strain slope is shaken mixing, with liquid-transfering gun respectively to be measured
Solution takes bacteria suspension and drips and be added drop-wise to respectively on stainless steel substrates, tinplate, aluminum-plastic composite membrane surface, room temperature in Biohazard Safety Equipment
Under, carry out experiment process after being dried 2h at once.
The conversion of uv dosage
Uv dosage is equal to the product of UV intensity with UV irradiation time, and concrete formula is as follows:
Uv dosage (J/cm2)=UV intensity (W/cm2) × exposure time (s)
Embodiment 1 peracetic acid processes the killing effect of Cronobacter sakazakii on different materials
By through sterilization treatment and pollute Cronobacter sakazakii and be dried stainless steel substrates, tinplate sheet and plastic-aluminum after 2 hours
Dripping concentration on composite membrane is the peracetic acid soln of 0.1%, 0.15%, 0.2%, 0.25% and 0.3%, after effect 10min
Utilize and stop agent termination reaction.Utilizing the bead of a diameter of 0.5mm that the bacterium of material business carries out eluting, and carrying out cultivating meter
Number.Peracetic acid is replaced to operate equally as blank using aseptic hard water.Before result of the test processes with peracetic acid
The Log value of rear material surface bacterium number decrement represents.Concrete outcome is as described in Table 1.
Table 1 variable concentrations peracetic acid is to the killing effect of Cronobacter sakazakii on different materials
Note: often in row, different expression of letter exists significant difference (P ＜ 0.05)
" ND " expression does not detects
As known from Table 1, the highest bactericidal effect of peroxyacetic acid concentration is the best.When concentration reaches 0.3%, Cronobacter sakazakii is not
It is detected.Simultaneously, it is known that different bacterial strains is different to the tolerance degree of peracetic acid on different materials.
Embodiment 2 chlorine dioxide treatment is to the killing effect of Cronobacter sakazakii on different materials
The present embodiment uses 500ppm, the chlorine dioxide of 1000ppm, 1500ppm, 2000ppm, 2500ppm, 3000ppm
Peracetic acid is replaced to have at the stainless steel substrates of four kinds of different Cronobacter sakazakii, tinplate sheet and aluminum-plastic composite membrane polluting
Reason, treatment conditions are identical with the condition of embodiment 1.Experimental result is as shown in table 2.
Table 2 variable concentrations chlorine dioxide is to the killing effect of Cronobacter sakazakii on different materials
As known from Table 2, with the increase of chlorine dioxide concentration, 4 strain bacterium decrement after treatment all gradually steps up, and works as dioxy
When change cl concn is 3000ppm, after process, through eluting, do not detect bacterium.4 strain bacterium are with 500ppm low concentration and 2500ppm
During higher chlorine dioxide treatment, through SPSS difference analysis, display all there was no significant difference (P between 4 strain bacterium on three kinds of materials
＞ 0.05).
Embodiment 3 hydrogen peroxide treatment is to the killing effect of Cronobacter sakazakii on different materials
The present embodiment uses 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, and the hydrogen peroxide of 3.5% replaces peracetic acid
Processing to pollute has the stainless steel substrates of four kinds of different Cronobacter sakazakii, tinplate sheet and aluminum-plastic composite membrane to process, treatment conditions
Identical with the condition of embodiment 1.Experimental result is as shown in table 3.
Table 3 variable concentrations hydrogen peroxide is to the killing effect of Cronobacter sakazakii on different materials
Note: often in row, different expression of letter exists significant difference (P ＜ 0.05)
" ND " expression does not detects.
As known from Table 3, along with the increase of concentration of hydrogen peroxide, 4 strain bacterium decrement of bacterium on three kinds of materials increases the most therewith
Add.When the concentration of hydrogen peroxide reaches 3.5%, four strain bacterium after treatment, all do not detect on three kinds of materials.In peroxidating
When the concentration of hydrogen is 3%, four strain bacterium there was no significant difference.
The different ultraviolet irradiation dose of embodiment 4 is to the killing effect of Cronobacter sakazakii on different materials
It is 0.1J/cm that the present embodiment have employed irradiation dose2、0.2J/cm2、0.3J/cm2、0.4J/cm2、0.5J/cm2's
Ultraviolet replaces peracetic acid to have the stainless steel substrates of four kinds of different Cronobacter sakazakii, tinplate sheet and plastic-aluminum combined to polluting
Film processes, and treatment conditions are identical with the condition of embodiment 1.Experimental result is as shown in table 4.
The different ultraviolet irradiation intensity of table 4 on different materials Cronobacter sakazakii do not kill effect
Note: often in row, different expression of letter exists significant difference (P ＜ 0.05)
" ND " expression does not detects
As known from Table 4, along with the increase of uv dosage, the bacterium of each ST type decrement on material is consequently increased.When
Uv dosage reaches 0.5J/cm2Time, material does not detects bacterium.When uv dosage is 0.1J/cm2、0.5J/cm2Time, three
Plant 4 strain bacterium on material all to there was no significant difference.When uv dosage is 0.2J/cm2、0.3J/cm2、0.4J/cm2Time, CE63 (ST1)
There is notable difference (P ＜ 0.05) with CE64 (ST4), and the bacterium number that CE63 (ST1) reduces is many, CE64 (ST4) and CE26
(ST268) between, difference is inconspicuous.
Embodiment 5 ultraviolet-disinfectant Combined Treatment insect killing effect to Cronobacter sakazakii
According to the result of embodiment 1-4, Cronobacter sakazakii is killed by the present embodiment by ultraviolet-disinfectant Combined Treatment
Effect is studied.The concentration of disinfectant is chosen for: the concentration of peracetic acid is 0.1%, 0.15%, and chlorine dioxide concentration is
500ppm, 1000ppm, concentration of hydrogen peroxide is 1.5%, 2.0%.Ultraviolet dosage chooses 0.2J/cm2、0.3J/cm2.At place
During reason, first spray disinfectant on the three kinds of materials polluted through Cronobacter sakazakii, after action time 30s, carry out ultraviolet
Line radiation treatment, is using aseptic waste water to carry out spray process after process, carries out the thalline of acquisition cultivating counting after process again.
Synergy is judged by below equation:
The bacterium number that synergistic action effect=Combined Treatment reduces-(the bacterium number that the bacterium number that ultraviolet reduces+disinfectant reduces) connection
Synergy analysis result after conjunction process, as shown in table 5.
Table 5 ultraviolet-disinfectant Combined Treatment insect killing effect to Cronobacter sakazakii
Note: " ND " expression does not detects
The bacterium number that synergistic action effect=Combined Treatment reduces-(the bacterium number that the bacterium number that ultraviolet reduces+disinfectant reduces) is
Represent during positive number and there is cooperative effect.During for negative, represent that the two has the phenomenon of fungicidal effectiveness cancellation.
As known from Table 5, when three kinds of disinfectant are combined with ultraviolet, all show positive cooperative effect, i.e. ratio two after combination
The bactericidal effect that person adds up after being used alone is good.There is obvious cooperative effect in ultraviolet and disinfectant combination, it is possible in both agent
When measuring relatively low, just produce a preferable sterilization impact.It is especially 2.0% at concentration of hydrogen peroxide, ultraviolet irradiation intensity
For 0.2J/cm2Time, cooperative effect is significantly higher than the technique effect of other groups.
Embodiment 6
According to the result of embodiment 5, inventor has turned down the intensity of ultraviolet irradiation and the consumption of disinfectant, uses disinfectant
Concentration is: the concentration of peracetic acid is 0.05%, and chlorine dioxide concentration is 400ppm, and concentration of hydrogen peroxide is 1.0%.Ultraviolet
Dosage chooses 0.1J/cm2Cronobacter sakazakii is processed.Result shows, UV treatment and three kinds of disinfectant Combined Treatment
The average synergistic action effect of four kinds of Cronobacter sakazakii is respectively tinplate: peracetic acid 0.11, chlorine dioxide 0.07, peroxide
Change hydrogen 0.26；Rustless steel: peracetic acid 0.12, chlorine dioxide 0.05, hydrogen peroxidase 10 .18；Aluminum-plastic composite membrane: peracetic acid
0.08, chlorine dioxide 0.09, hydrogen peroxidase 10 .34.Synergistic action effect is significantly lower than embodiment 5.
Embodiment 7
According to the result of embodiment 5, it is 0.4J/cm that inventor heightens the level of ultraviolet irradiation dose2, the use of disinfectant
Amount level is: the concentration of peracetic acid is 0.1%, and chlorine dioxide concentration is 500ppm, and concentration of hydrogen peroxide is 1.5%.Utilize
Cronobacter sakazakii is processed by above-mentioned condition (other conditions are same as in Example 5).Result shows, peracetic acid, titanium dioxide
Chlorine and three kinds of disinfectant of hydrogen peroxide are used in combination the average synergy to four kinds of Cronobacter sakazakii with ultraviolet and are followed successively by: horse
Mouth ferrum: peracetic acid 0.89, chlorine dioxide 0.58, hydrogen peroxidase 10 .77；Rustless steel: peracetic acid 0.78, chlorine dioxide 0.33,
Hydrogen peroxide 1.11；Aluminum-plastic composite membrane: peracetic acid 0.88, chlorine dioxide 0.51, hydrogen peroxide 1.23.From the above may be used
Knowing, synergistic effect does not improve, the most generally less than the result of embodiment 5 cooperative effect.
Although the present invention is open the most as above with preferred embodiment, but it is not limited to the present invention, any is familiar with this
The people of technology, without departing from the spirit and scope of the present invention, can do various change and modification, the therefore protection of the present invention
Scope should be with being as the criterion that claims are defined.
Claims (6)
1. the physical chemistry combined sterilizing method of a Cronobacter sakazakii, it is characterised in that spray sterilization on object subject to sterilization
Agent, after disinfectant effect 20-40s, utilizes ultraviolet to carry out radiation treatment, and irradiation dose reaches 0.2-0.3J/cm2After, stop
Ultraviolet radiation treatment also utilizes aseptic clear water to spray object subject to sterilization；Described disinfectant is peracetic acid, chlorine dioxide or mistake
Hydrogen oxide.
Method for disinfection the most according to claim 1, it is characterised in that step is as follows:
1) in object subject to sterilization, spray peracetic acid, chlorine dioxide or hydrogenperoxide steam generator, act on 20-40s；
2) to step 1) pass through the object subject to sterilization disinfected to carry out irradiation dose being 0.2-0.3J/cm2Ultraviolet irradiation at
Reason；
3) aseptic clear water object subject to sterilization to cleaning showers is utilized.
Method for disinfection the most according to claim 2, it is characterised in that step 1) described object subject to sterilization is that plant produced sets
Standby or packaging material.
Method for disinfection the most according to claim 2, it is characterised in that the material of described object subject to sterilization is rustless steel, horse
Mouth ferrum, aluminum-plastic composite membrane, glass or cement.
Method for disinfection the most according to claim 2, it is characterised in that step 1) mass fraction of described peracetic acid soln
For 0.1-0.15%；Described chlorine dioxide addition is 500-1000ppm；The mass fraction of described hydrogen peroxide is 1.5-
2.0%.
6. claim 1-5 arbitrary described method application during the sterilization of food manufacturing apparatus and packaging material.
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| CN106039353B (en) | 2019-01-25 |
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