WO2011159054A2 - Method for identifying proper storage of goods requiring storage at low temperature - Google Patents

Method for identifying proper storage of goods requiring storage at low temperature Download PDF

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Publication number
WO2011159054A2
WO2011159054A2 PCT/KR2011/004166 KR2011004166W WO2011159054A2 WO 2011159054 A2 WO2011159054 A2 WO 2011159054A2 KR 2011004166 W KR2011004166 W KR 2011004166W WO 2011159054 A2 WO2011159054 A2 WO 2011159054A2
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marker
storage
low temperature
microorganisms
indication
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PCT/KR2011/004166
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French (fr)
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WO2011159054A3 (en
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Sang Ryoul Park
Sook-Kyung Kim
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Korea Research Institute Of Standards And Science
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Publication of WO2011159054A3 publication Critical patent/WO2011159054A3/en

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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor

Definitions

  • the present invention relates to a method for identifying proper storage of goods requiring storage at a low temperature in which proper storage of products or goods requiring storage at a low temperature may be easily identified.
  • An object of the present invention is to economically implement and widely utilize an indication marker capable of easily identifying with a naked eye whether goods (for example, foods, medical supplies, high class chemical reagents, biological materials, or the like) requiring storage at a low temperature are properly stored in a low temperature state as defined.
  • Another object of the present invention is to provide a method for identifying proper storage of goods requiring storage at a low temperature.
  • a method for identifying proper storage of goods requiring storage at a low temperature in which a culture medium including an indicator and marker microorganisms is mounted, as an indication marker, in the goods requiring storage at a low temperature, such that growth of the marker microorganisms is controlled by a storage condition, and proper storage of the goods requiring storage at a low temperature is identified according to a degree in which a color of the indication marker is changed according to the growth of the marker microorganisms.
  • the color of the indication marker may be changed due to increase in metabolic products according to the growth of the marker microorganisms.
  • the color of the indication marker may be changed due to reduction in metabolic reactants of the culture medium according to the growth of the marker microorganisms.
  • the color of the indication marker may be changed due to a change in pH of the culture medium according to the growth of the marker microorganisms.
  • the indicator may include all indicators capable of confirming a change in color according to the growth of the marker microorganisms, preferably, Resorcinol blue (Lacmoid).
  • the Resorcinol blue may be a coloring indicator for indicating pH and allow a change in pH to cause a remarkable change in color, such that it may be a preferable example of the indicator.
  • pH buffer capacity of the culture medium is controlled, thereby making it possible to adjust serious deviation from the required storage condition at a low temperature and a rapid change in pH so as to coincide with each other at maiximum.
  • the marker microorganisms may include bacteriomycota selected from Lactobacillus sp, Leuconostoc sp, Pediococcus.
  • the culture medium may be a modified MRS medium.
  • the marker- microorganisms Before the indication marker is mounted on the goods requiring storage at a low temperature, the marker- microorganisms may be separated from the culture medium by a separating film, and when it is mounted on the goods, the separating film may be damaged, such that the marker microorganisms is mixed with the culture medium.
  • the indicator When the marker microorganisms is separated from the culture medium by a separating film, the indicator may be pre-mixed with the culture medium for uniform distribution in the medium.
  • the indication marker may have a volume of 1 to 1000 ⁇ l. When the indication maker is a hydrogel type, damage possibility of the indication marker may be reduced.
  • a method for identifying proper storage of goods requiring storage at a low temperature in which whether the goods requiring storage at a low temperature have been properly stored may be easily identified with a naked eye by accumulating a situation in which the goods deviate from a proper low temperature storage state whenever the goods requiring storage at a low temperature deviates from the proper low temperature storage state is provided in the present invention.
  • the method for identifying proper storage of goods requiring storage at a low temperature is characterized in that whether the goods requiring storage at a low temperature are properly stored at a low temperature may be indentified according to a growth degree of the marker microorganisms by using a fact that the marker microorganisms properly receiving nutritive elements from the culture medium, in an indication marker including a culture medium including an indicator and marker microorganisms are actively grown when they are repeatedly or continuously exposed at a temperature higher than a specific temperature.
  • the method for identifying proper storage of goods requiring storage at a low temperature may become an excellent method for recording a storage state of the goods.
  • the goods requiring storage at a low temperature may have different conditions for proper low temperature storage according to the goods.
  • the conditions may be controlled by adjusting a kind of marker microorganisms and a supply degree of nutritive elements through a medium.
  • microorganisms developed by using modification of species such as genetic recombination, or the like, according to required conditions may be used as the marker microorganisms.
  • species harmless to human and environment may be selected.
  • microorganisms reflecting whether or not different storage conditions for each of application goods are satisfied may be selected and used through a test.
  • the growth degree of the marker microorganisms may be identified through a change in color of an indication marker due to reduction in metabolic reactants at the time of deviation from low temperature storage state, coloring due to increase in metabolic products according to growth of the marker microorganisms, and a change in color of the indication marker caused by a chemical reaction, in a provided medium.
  • active growth of the marker microorganisms may be recognized from rapid reduction in a concentration of a material required for metabolism of the marker microorganisms or rapid increase in a concentration of the metabolic products of the corresponding marker microorganisms.
  • the metabolic reactants are rapidly reduced or metabolic products are rapidly increased due to the rapid growth of the marker microorganisms to cause a color change reaction of the indication marker, thereby rapidly changing a color of a culture medium.
  • pH of the culture medium is lowered.
  • a predetermined coloring indicator for indicating pH is added to the culture medium, such that a change in color of the culture medium may be generated so as to be identifiable with a naked eye.
  • a small amount of the indication marker which is a volume of 1 to 1000 ⁇ l, may be mounted in the goods.
  • a container including the indication marker may be made of a transparent material so that a change in color may be easily found, more specifically, a plastic material.
  • the container including the indication marker may have a flat shape in order to prevent damage of the goods requiring storage at a low temperature and in order not to incur an inconvenience in loading the goods at the time of carrying the goods.
  • the container including the indication marker may have an outer bottom surface treated with an adhesive so that the container is properly attached to the goods requiring storage at a low temperature.
  • a separating film of the indication marker is processed to be punctured by intentionally applied impact, and may be made of any material as long as it is a material that does not have an influence on growth of microorganisms, without being specifically limited.
  • a method for completely puncturing the separating film by previously making a guideline for puncture in the separating film, attaching the separating film, and then intentionally applying impact to the separating film, thereby mixing the culture medium with the marker microorganisms, may also be used.
  • the separating film may be made of low density polyethylene.
  • the culture medium may be separated from the marker microorganisms by the separating film before the time point at which storage at a low temperature is required and the separating film may be intentionally damaged at the time point at which storage at a low temperature is required to thereby allow the marker microorganisms to enter the culture medium including the indicator, such that recording with respect to a low temperature storage state starts from the time point at which storage at a low temperature is required.
  • goods for which low temperature storage management is not properly made leads to a clear change in color of the indication marker, such that they may be easily recognized by everyone.
  • the indication marker may be used in all kinds of goods continuously requiring storage at a low temperature during a distribution period, a transport period, or a use period and may be attached to a minimal package unit or a larger package unit of goods that are expected to individually distributed, thereby identifying whether or not a storage state is proper during the processes of storage, distribution, and use of the goods in each unit.
  • the goods capable of causing significant deterioration of marketability or a health hazard due to a change in quality when low temperature storage is not thoroughly made may be effectively managed.
  • the indication marker according to the present invention may be implemented at a low cost and recognize improperly stored goods with a naked eye without requiring a separate specific tool, such that it may be widely applied in various products.
  • FIG. 1 is a view showing a method for identifying proper storage of goods requiring storage at a low temperature according to the present invention
  • FIG. 2 is a graph showing absorbance spectra immediately after a medium including an indicator and a separating film in Example 1 is damaged;
  • FIG. 3 is a graph showing absorbance spectra after a separating film of an indication marker produced by Example 1 is damaged and the indication marker is then exposed at a room temperature for 15 hours;
  • FIG. 4 is a graph showing absorbance spectra after an indication marker produced by Example 1 is exposed at a room temperature for 60 minutes every three hours six times.
  • FIG. 1 is a view showing a method for identifying proper storage of goods requiring storage at a low temperature according to the present invention.
  • An indication marker according to Fig 1 includes a marker microorganism, a culture medium, and a pH-responsive coloring indicator. Test conditions applied in this test are as follows.
  • a test indicater solution of the final volume of 100 ⁇ l was composed by the following method. Leuconostoc mesenteroides subsp and Leuconostoc mesenteroides KCTC 3505 were used as the marker microorganism. Firstly, 10 ⁇ l of a preparation of the marker microorganism (about 10 7 CFU/ml) was taken, so that its final concentration should become about 10 6 CFU/ml when mixed with the rest of the test indicater soultion. For preaparation of the rest of the test indicater solution, 50 ⁇ l of a modified MRS medium of was taken, such that its final concentration became 1/2.

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Abstract

Provided is a method for identifying proper storage of goods requiring storage at a low temperature in which a culture medium including an indicator and marker microorganisms is mounted, as an indication marker, in the goods requiring storage at a low temperature, such that growth of the marker microorganisms is controlled by a storage condition, and proper storage of the goods requiring storage at a low temperature is identified according to a degree in which a color of the indication marker is changed according to the growth of the marker microorganisms.

Description

METHOD FOR IDENTIFYING PROPER STORAGE OF GOODS REQUIRING STORAGE AT LOW TEMPERATURE
The present invention relates to a method for identifying proper storage of goods requiring storage at a low temperature in which proper storage of products or goods requiring storage at a low temperature may be easily identified.
It is very important in a distribution process of goods requiring storage in a cold or refrigerated state to confirm that the goods are properly stored during processes of storage, distribution, and use thereof. However, technologies capable of easily identifying with a naked eye whether goods requiring storage at a low temperature are properly stored in a low temperature state as defined have not been developed.
An object of the present invention is to economically implement and widely utilize an indication marker capable of easily identifying with a naked eye whether goods (for example, foods, medical supplies, high class chemical reagents, biological materials, or the like) requiring storage at a low temperature are properly stored in a low temperature state as defined. Another object of the present invention is to provide a method for identifying proper storage of goods requiring storage at a low temperature.
In one general aspect, there is provided a method for identifying proper storage of goods requiring storage at a low temperature in which a culture medium including an indicator and marker microorganisms is mounted, as an indication marker, in the goods requiring storage at a low temperature, such that growth of the marker microorganisms is controlled by a storage condition, and proper storage of the goods requiring storage at a low temperature is identified according to a degree in which a color of the indication marker is changed according to the growth of the marker microorganisms.
The color of the indication marker may be changed due to increase in metabolic products according to the growth of the marker microorganisms. The color of the indication marker may be changed due to reduction in metabolic reactants of the culture medium according to the growth of the marker microorganisms. The color of the indication marker may be changed due to a change in pH of the culture medium according to the growth of the marker microorganisms.
The indicator may include all indicators capable of confirming a change in color according to the growth of the marker microorganisms, preferably, Resorcinol blue (Lacmoid). The Resorcinol blue may be a coloring indicator for indicating pH and allow a change in pH to cause a remarkable change in color, such that it may be a preferable example of the indicator. When the coloring indicator for indicating pH is used, pH buffer capacity of the culture medium is controlled, thereby making it possible to adjust serious deviation from the required storage condition at a low temperature and a rapid change in pH so as to coincide with each other at maiximum.
The marker microorganisms may include bacteriomycota selected from Lactobacillus sp, Leuconostoc sp, Pediococcus. The culture medium may be a modified MRS medium.
Before the indication marker is mounted on the goods requiring storage at a low temperature, the marker- microorganisms may be separated from the culture medium by a separating film, and when it is mounted on the goods, the separating film may be damaged, such that the marker microorganisms is mixed with the culture medium. When the marker microorganisms is separated from the culture medium by a separating film, the indicator may be pre-mixed with the culture medium for uniform distribution in the medium. The indication marker may have a volume of 1 to 1000 ㎕. When the indication maker is a hydrogel type, damage possibility of the indication marker may be reduced.
Hereinafter, the present invention will be described in more detail.
A method for identifying proper storage of goods requiring storage at a low temperature in which whether the goods requiring storage at a low temperature have been properly stored may be easily identified with a naked eye by accumulating a situation in which the goods deviate from a proper low temperature storage state whenever the goods requiring storage at a low temperature deviates from the proper low temperature storage state is provided in the present invention.
The method for identifying proper storage of goods requiring storage at a low temperature is characterized in that whether the goods requiring storage at a low temperature are properly stored at a low temperature may be indentified according to a growth degree of the marker microorganisms by using a fact that the marker microorganisms properly receiving nutritive elements from the culture medium, in an indication marker including a culture medium including an indicator and marker microorganisms are actively grown when they are repeatedly or continuously exposed at a temperature higher than a specific temperature. Since the growth degree of the marker microorganisms accumulatively reflects the exposure of the marker microorganisms at a temperature of a proper temperature or more, the method for identifying proper storage of goods requiring storage at a low temperature according to the present invention may become an excellent method for recording a storage state of the goods.
In addition, the goods requiring storage at a low temperature may have different conditions for proper low temperature storage according to the goods. The conditions may be controlled by adjusting a kind of marker microorganisms and a supply degree of nutritive elements through a medium. In the present invention, microorganisms developed by using modification of species such as genetic recombination, or the like, according to required conditions may be used as the marker microorganisms. For the marker microorganisms species harmless to human and environment may be selected. In addition, microorganisms reflecting whether or not different storage conditions for each of application goods are satisfied may be selected and used through a test.
The growth degree of the marker microorganisms may be identified through a change in color of an indication marker due to reduction in metabolic reactants at the time of deviation from low temperature storage state, coloring due to increase in metabolic products according to growth of the marker microorganisms, and a change in color of the indication marker caused by a chemical reaction, in a provided medium.
More specifically, active growth of the marker microorganisms may be recognized from rapid reduction in a concentration of a material required for metabolism of the marker microorganisms or rapid increase in a concentration of the metabolic products of the corresponding marker microorganisms. According to the present invention, the metabolic reactants are rapidly reduced or metabolic products are rapidly increased due to the rapid growth of the marker microorganisms to cause a color change reaction of the indication marker, thereby rapidly changing a color of a culture medium.
More specifically, when the marker microorganisms generate an acid material as the metabolic products at the time of deviation from low temperature storage state, pH of the culture medium is lowered. At this time, a predetermined coloring indicator for indicating pH is added to the culture medium, such that a change in color of the culture medium may be generated so as to be identifiable with a naked eye.
A small amount of the indication marker, which is a volume of 1 to 1000 ㎕, may be mounted in the goods. A container including the indication marker may be made of a transparent material so that a change in color may be easily found, more specifically, a plastic material. The container including the indication marker may have a flat shape in order to prevent damage of the goods requiring storage at a low temperature and in order not to incur an inconvenience in loading the goods at the time of carrying the goods. In addition, in order to easily attach the indication marker, the container including the indication marker may have an outer bottom surface treated with an adhesive so that the container is properly attached to the goods requiring storage at a low temperature. A separating film of the indication marker is processed to be punctured by intentionally applied impact, and may be made of any material as long as it is a material that does not have an influence on growth of microorganisms, without being specifically limited. In addition, a method for completely puncturing the separating film by previously making a guideline for puncture in the separating film, attaching the separating film, and then intentionally applying impact to the separating film, thereby mixing the culture medium with the marker microorganisms, may also be used. More preferably, the separating film may be made of low density polyethylene.
In addition, in the indication marker, the culture medium may be separated from the marker microorganisms by the separating film before the time point at which storage at a low temperature is required and the separating film may be intentionally damaged at the time point at which storage at a low temperature is required to thereby allow the marker microorganisms to enter the culture medium including the indicator, such that recording with respect to a low temperature storage state starts from the time point at which storage at a low temperature is required. According to the present invention, goods for which low temperature storage management is not properly made leads to a clear change in color of the indication marker, such that they may be easily recognized by everyone. When the indicator is separated from the marker microorganisms and is then mixed with the culture medium, it may be uniformly spread to the indication marker.
The indication marker may be used in all kinds of goods continuously requiring storage at a low temperature during a distribution period, a transport period, or a use period and may be attached to a minimal package unit or a larger package unit of goods that are expected to individually distributed, thereby identifying whether or not a storage state is proper during the processes of storage, distribution, and use of the goods in each unit.
According to the present invention, the goods capable of causing significant deterioration of marketability or a health hazard due to a change in quality when low temperature storage is not thoroughly made may be effectively managed. The indication marker according to the present invention may be implemented at a low cost and recognize improperly stored goods with a naked eye without requiring a separate specific tool, such that it may be widely applied in various products.
The above and other objects, features and advantages of the present invention will become apparent from the following description of preferred embodiments given in conjunction with the accompanying drawings, in which:
FIG. 1 is a view showing a method for identifying proper storage of goods requiring storage at a low temperature according to the present invention;
FIG. 2 is a graph showing absorbance spectra immediately after a medium including an indicator and a separating film in Example 1 is damaged;
FIG. 3 is a graph showing absorbance spectra after a separating film of an indication marker produced by Example 1 is damaged and the indication marker is then exposed at a room temperature for 15 hours; and
FIG. 4 is a graph showing absorbance spectra after an indication marker produced by Example 1 is exposed at a room temperature for 60 minutes every three hours six times.
Hereinafter, a detailed description of the present invention will be provided by way of example with reference to the accompanying drawings. However, the present invention is not limited to the following Example. FIG. 1 is a view showing a method for identifying proper storage of goods requiring storage at a low temperature according to the present invention.
[Example 1]
An indication marker according to Fig 1 includes a marker microorganism, a culture medium, and a pH-responsive coloring indicator. Test conditions applied in this test are as follows.
A test indicater solution of the final volume of 100 ㎕ was composed by the following method. Leuconostoc mesenteroides subsp and Leuconostoc mesenteroides KCTC 3505 were used as the marker microorganism. Firstly, 10 ㎕ of a preparation of the marker microorganism (about 107 CFU/ml) was taken, so that its final concentration should become about 106 CFU/ml when mixed with the rest of the test indicater soultion. For preaparation of the rest of the test indicater solution, 50 ㎕ of a modified MRS medium of was taken, such that its final concentration became 1/2. 1 ㎕ of the pH-responsive coloring indicator (Resorcinol blue or Lacmoid) prepared at a concentration of 1 % was added to the modified MRS medium, such that its final concentration became 0.01 %. Finally, 39 ㎕ of deionized water was added to the mixture of the coloring indicater and the modified MRS medium, such that the volume of the rest of the test indicater solution was 90㎕ that was initially separated from the indicater microorganism by a thin film as proposed by Fig 1.
The rest of the test indicater solution produced as Example 1 was measured pH 6.2 and exhibited a blue color. Absorption spectra at this stageis shown in FIG. 2 (a graph of a the test indicater solution witout the indicater microorganismin FIG. 2). Absorption spectra iniucater solutionin FIG. 2) immediately after damaging the separating film (a graph of the whole test indicater solution not exposed to a high temperature) is also shown in FIG. 2. It may be appreciated that an absorbance curve immediately after the whole test indicater solution is produced or in the state of low temperature storage exhibits a blue color or a blue purple color.
When a separating film of an indication marker produced by Example 1 is damaged and the indication marker is then exposed at a room temperature for 15 hours, the indication maker exhibited a red color. At this time, pH was 4.0 and the observed absorption spectra is shown in FIG. 3.
In the case of continuously storing the indication marker produced by Example 1 at a temperature of 4 , a change in color was not found at all and pH was maintained at 6.0 or more.
In the case of exposing an indication marker of 100 ㎕ produced by Example 1 at a room temperature for 60 minutes every three hours six times, a change in color started to be found. At the time point in which the change in color starts, pH was 4.7 and absorbance spectra was shown in FIG. 4. As a result of the change in color, a red color was exhibited.

Claims (9)

  1. A method for identifying proper storage of goods requiring storage at a low temperature in which a culture medium including a marker microorganisms and an indicator is mounted, as an indication marker, in the goods requiring storage at a low temperature, such that growth of the marker microorganisms is controlled by a storage condition, and proper storage of the goods requiring storage at a low temperature is identified according to a degree in which a color of the indication marker is changed according to the growth of the marker microorganisms.
  2. The method of claim 1, wherein the color of the indication marker is changed due to increase in metabolic products according to the growth of the marker microorganisms.
  3. The method of claim 1, wherein the color of the indication marker is changed due to reduction in metabolic reactants of the culture medium according to the growth of the marker microorganisms.
  4. The method of claim 1, wherein the color of the indication marker is changed due to a change in pH of the culture medium according to the growth of the marker microorganisms.
  5. The method of claim 4, wherein the indicator is Resorcinol blue (Lacmoid).
  6. The method of claim 1, wherein the marker microorganisms include bacteriomycota selected from Lactobacillus sp, Leuconostoc sp, Pediococcus.
  7. The method of claim 6, wherein the culture medium is a modified MRS medium.
  8. The method of claim 1, wherein before the indication marker is mounted in the goods requiring storage at a low temperature, the marker microorganisms is separated from the culture medium by a separating film, and when it is mounted in the goods requiring storage at a low temperature, the separating film is damaged, such that the marker microorganisms is mixed with the culture medium.
  9. The method of claim 8, wherein the indication marker has a volume of 1 to 1000 ㎕ and is a hydrogel type.
PCT/KR2011/004166 2010-06-16 2011-06-08 Method for identifying proper storage of goods requiring storage at low temperature WO2011159054A2 (en)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20020066157A (en) * 2001-02-09 2002-08-14 박찬훈 A bio-indicator for monitoring of microbial decomposition
US20070275467A1 (en) * 2003-09-17 2007-11-29 Cryolog S.A. Method and Device for Determining if a Product is in Condition for Use or Consumption
KR20080005975A (en) * 2005-04-25 2008-01-15 에스쥐엠 바이오텍, 인크. Evaluating bacterial lethality of containerized food production
KR20100011202A (en) * 2008-07-24 2010-02-03 인하대학교 산학협력단 Label for estimating of saprogenic bacteria's growth by cumulative storage temperature' change

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20020066157A (en) * 2001-02-09 2002-08-14 박찬훈 A bio-indicator for monitoring of microbial decomposition
US20070275467A1 (en) * 2003-09-17 2007-11-29 Cryolog S.A. Method and Device for Determining if a Product is in Condition for Use or Consumption
KR20080005975A (en) * 2005-04-25 2008-01-15 에스쥐엠 바이오텍, 인크. Evaluating bacterial lethality of containerized food production
KR20100011202A (en) * 2008-07-24 2010-02-03 인하대학교 산학협력단 Label for estimating of saprogenic bacteria's growth by cumulative storage temperature' change

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