CN109666590A - A kind of method of microorganism in collection fresh-cut fruit and vegetable - Google Patents
A kind of method of microorganism in collection fresh-cut fruit and vegetable Download PDFInfo
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- CN109666590A CN109666590A CN201910086839.4A CN201910086839A CN109666590A CN 109666590 A CN109666590 A CN 109666590A CN 201910086839 A CN201910086839 A CN 201910086839A CN 109666590 A CN109666590 A CN 109666590A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/02—Separating microorganisms from the culture medium; Concentration of biomass
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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Abstract
The present invention relates to field of detection of food safety, and in particular, to a method of collect microorganism in fresh-cut fruit and vegetable.Listeria monocytogenes, Escherichia coli O 157: H7, staphylococcus aureus are subjected to activation culture, respectively obtain the bacterium solution of three kinds of bacterium, sterile working takes fresh fruit or vegetables, 9-11g sample is weighed after cutting, respectively according to three kinds of pathogenic bacteria of 8-9log cfu/mL artificial contamination, after drying up 0.5-1.5h in aseptic operating platform, it is put into the sterile homogenizing bag containing 85-95mL 0.1wt% peptone water, it pats and mixes on homogenizer, fruits and vegetables homogenizing fluid is fallen in Double-filtering device, microorganism is deposited on filter device.For the present invention in fresh-cut fruit and vegetable product when microorganism detection, a step collects whole microorganisms in fruit and vegetable food, time saving and energy saving, avoids pollution.
Description
Technical field
The present invention relates to field of detection of food safety, and in particular, to a method of collect microorganism in fresh-cut fruit and vegetable.
Background technique
Food-borne pathogens are the principal element for endangering food safety and human health, Listeria monocytogenes, Escherichia coli
O157:H7 and Staphylococcus aureus belong to infecting both domestic animals and human pathogenic microorganism, and widely distributed in nature, clinical manifestation
For symptoms such as Nausea and vomiting, abdominal pain, diarrhea.Fresh-cut fruit and vegetable is easily infected by it in storage with transportational process, and to cause of disease
During microorganism detection, fruits and vegetables can generate a large amount of fruits and vegetables residue and small fruit after homogenizer is patted,
Different degrees of interference can be brought for the counting of subsequent microbe colony, PCR detection etc., to influence the accuracy of testing result.
Molecule containing fruits and vegetables in the microorganism that the existing method for collecting microorganism is collected, the particles effect
The progress of subsequent experimental.
Summary of the invention
In view of the deficiencies of the prior art, the present invention provides a kind of method for collecting microorganism in fresh-cut fruit and vegetable, and this method can
To avoid being mixed into for the molecule in fruits and vegetables during collecting microorganism.
The present invention solves the technical solution that technical problem uses are as follows:
By Listeria monocytogenes, Escherichia coli O 157: H7, staphylococcus aureus carry out activation culture, respectively obtain three
The bacterium solution of kind bacterium, sterile working take fresh fruit or vegetables, weigh 9-11g sample, manually dirty according to 8-9log cfu/mL respectively
Three kinds of pathogenic bacteria are contaminated, after drying up 0.5-1.5h in aseptic operating platform, are put into the nothing containing 85-95mL 0.1wt% peptone water
It in bacterium homogenizing bag, pats and mixes on homogenizer, fruits and vegetables homogenizing fluid is fallen in Double-filtering device, microorganism is deposited in filtering dress
It sets.
The Double-filtering device includes: fruits and vegetables homogeneous collection, large aperture filter membrane, bacterium solution collector, aperture
Diameter filter membrane and filtrate displacer.Wherein fruits and vegetables homogeneous collection is cylindric hollow structure, and the diameter of two bottom surfaces is greater than
The diameter of cylinder, two bottom edges protrude from cylindrical sides, and it is identical as column diameter big that area is removed in two bottom center positions
Small circle forms hollow cylinder, the structure of side is protruded from around two bottoms, and be set to top in ground according to central axis,
Fruits and vegetables homogeneous collection bottom end is equipped with bacterium solution collector, and bacterium solution collector and fruits and vegetables homogeneous collection shape, size are complete
It is identical, large aperture filter membrane, the area of large aperture filter membrane are equipped between fruits and vegetables homogeneous collection and bacterium solution collector top
Identical as the base area of fruits and vegetables homogeneous collection, bacterium solution collector bottom end is equipped with filtrate displacer, bacterium solution collector bottom end
Small-bore filter membrane is equipped between filtrate displacer, the area of small-bore filter membrane is identical as bacterium solution collector bottom end area,
Filtrate displacer upper end is identical as the bottom shape of fruits and vegetables homogeneous collection, and filtrate displacer sub-body is that diameter is less than filter
The hollow pipe of liquid displacer basal diameter, the hollow pipe lower end are beveling shape.
The material of the fruits and vegetables homogeneous collection, bacterium solution collector and filtrate displacer is glass material, each section
It clamps protrusion by clip to be fixed, fruits and vegetables homogeneous collection, bacterium solution collector, filtrate displacer, large aperture mistake
It is to be detachably connected between filter membrane and small-bore filter membrane.
The large aperture filter membrane is polypropylene or nylon material, and the material of small-bore filter membrane is nylon material.
When large aperture filter membrane is polypropylene screen, pore size filter range is 10 μm -40 μm, and large aperture filter membrane is nylon mistake
When filter membrane, pore diameter range is 15 μm -60 μm；The pore size filter range of small-bore filter membrane is 0.22 μm -0.45 μm.
The beneficial effects of the present invention are: one step collects fruit and vegetable food in fruits and vegetables product when microorganism detection
Middle whole microorganism, it is time saving and energy saving, it avoids pollution.The present invention overcomes can not completely, quickly collect microorganism in fruit and vegetable food
Difficulty, while avoiding fruits and vegetables residue to the subsequent influence for Molecular Detection.
Detailed description of the invention
Fig. 1 is the structure chart of Double-filtering device.
Fig. 2 is the top view of large aperture filter membrane.
Fig. 3 is the top view of small-bore filter membrane.
Such as Fig. 1 fruits and vegetables homogeneous collection, 2. large aperture filter membranes, 3. bacterium solution collectors, 4. small-bore filter membranes, 5.
Filtrate displacer.
Specific embodiment
Below with reference to examples and drawings, detailed description further is made to invention, but embodiments of the present invention are not
It is limited to this.
Embodiment 1
By Listeria monocytogenes, Escherichia coli O 157: H7, staphylococcus aureus carry out activation culture, respectively obtain three
The bacterium solution of kind bacterium, sterile working take fresh "Hami" melon sample, by its cutting, 10g sample are weighed, respectively according to 9log cfu/mL
Three kinds of pathogenic bacteria of artificial contamination are put into after drying up 1h in aseptic operating platform containing the sterile of 95mL 0.1wt% peptone water
It in homogenizing bag, pats and mixes on homogenizer, "Hami" melon homogenizing fluid is fallen in Double-filtering device, wherein large aperture filter membrane 2
(polypropylene filter film, 10,20,40 μm of pore size) can intercept the residue after "Hami" melon homogeneous, and the filtrate with microorganism is logical
Bacterium solution collector 3 is crossed, small-bore filter membrane 4 (nylon filter film, pore size are 0.22 μm and 0.45 μm) collects all filters
The microorganism crossed, filtrate filter through filtered fluid displacer 5.The small-bore filter membrane 4 of attached microbial can be removed, and can be carried out subsequent
Microorganism detection.
Embodiment 2
By Listeria monocytogenes, Escherichia coli O 157: H7, staphylococcus aureus carry out activation culture, respectively obtain three
The bacterium solution of kind bacterium, sterile working take fresh "Hami" melon sample, by its cutting, 10g sample are weighed, respectively according to 9log cfu/mL
Three kinds of pathogenic bacteria of artificial contamination are put into after drying up 1h in aseptic operating platform containing the sterile of 90mL 0.1wt% peptone water
It in homogenizing bag, pats and mixes on homogenizer, "Hami" melon homogenizing fluid is fallen in Double-filtering device, 2 (Buddhist nun of large aperture filter membrane
Imperial filter membrane, 15,40,60 μm of pore size) residue after "Hami" melon homogeneous can be intercepted, the filtrate with microorganism passes through bacterium solution
Collector 3, small-bore filter membrane 4 (nylon filter film, pore size be 0.22 μm and 0.45 μm) is collectable all filter it is micro-
Biology, filtrate filter through filtered fluid displacer.The small-bore filter membrane 4 of attached microbial can be removed, and can carry out subsequent microorganism
Detection.
Embodiment 3
By Listeria monocytogenes, Escherichia coli O 157: H7, staphylococcus aureus carry out activation culture, respectively obtain three
The bacterium solution of kind bacterium, sterile working take fresh Lettuce samples, by its cutting, 10g sample are weighed, respectively according to 9log cfu/mL people
Work pollutes three kinds of pathogenic bacteria, after drying up 1h in aseptic operating platform, is put into sterile equal containing 90mL 0.1wt% peptone water
It in matter bag, pats and mixes on homogenizer, its homogenizing fluid is fallen in Double-filtering device, 2 (nylon filter of large aperture filter membrane
Film, 15,40,60 μm of pore size) residue after lettuce homogeneous can be intercepted, the filtrate with microorganism passes through bacterium solution collector 3,
Small-bore filter membrane 4 (nylon filter film, pore size are 0.22 μm and 0.45 μm) collects the microorganism all filtered, filtrate
It is filtered through filtered fluid displacer.The small-bore filter membrane 4 of attached microbial can be removed, and can carry out subsequent microorganism detection.
Embodiment 4
By Listeria monocytogenes, Escherichia coli O 157: H7, staphylococcus aureus carry out activation culture, respectively obtain three
The bacterium solution of kind bacterium, sterile working take fresh Lettuce samples, by its cutting, 10g sample are weighed, respectively according to 9log cfu/mL people
Work pollutes three kinds of pathogenic bacteria, after drying up 1h in aseptic operating platform, is put into sterile equal containing 90mL 0.1wt% peptone water
It in matter bag, pats and mixes on homogenizer, lettuce homogenizing fluid is fallen in Double-filtering device, (the nylon mistake of large aperture filter membrane 2
Filter membrane, 15,40,60 μm of pore size) residue after lettuce homogeneous can be intercepted, the filtrate with microorganism passes through bacterium solution collector
3, small-bore filter membrane 4 (nylon filter film, pore size are 0.22 μm and 0.45 μm) collects the microorganism all filtered, filter
Liquid is filtered through filtered fluid displacer.The small-bore filter membrane 4 of attached microbial can be removed, and can carry out subsequent microorganism detection.
As shown in table 1,2,3,4, table 1 is for embodiment 1, embodiment 2, embodiment 3, the microorganism detection result of embodiment 4
The testing result of embodiment 1, table 2 are the testing result of embodiment 2, and table 3 is the testing result of embodiment 3, table 4 is embodiment 4
Testing result.
1 pathogenic microorganism of table is inoculated in the clump count filtered after fresh-cut "Hami" melon through polypropylene screen
2 pathogenic microorganism of table is inoculated in the clump count filtered after fresh-cut "Hami" melon through nylon membrane
3 pathogenic microorganism of table is inoculated in the clump count filtered after fresh-cut water chestnut through polypropylene screen
4 pathogenic microorganism of table is inoculated in the clump count filtered after fresh-cut water chestnut through nylon membrane
Claims (4)
1. a kind of method for collecting microorganism in fresh-cut fruit and vegetable, which is characterized in that this method operating procedure is as follows:
S1. by Listeria monocytogenes, Escherichia coli O 157: H7, staphylococcus aureus carry out activation culture, respectively obtain three kinds
The bacterium solution of bacterium；
S2. sterile working takes fresh fruit or vegetables, by its cutting, weighs 9-11g；
S3. three kinds of pathogenic bacteria of fruit or vegetables artificial contamination into step S2；
S4. it after drying up 0.5-1.5h in aseptic operating platform, is put into the sterile homogenizing bag containing peptone water, on homogenizer
It pats and mixes；
S5. fruits and vegetables homogenizing fluid is fallen in Double-filtering device, microorganism is deposited on filter device.
2. a kind of method for collecting microorganism in fresh-cut fruit and vegetable according to claim 1, which is characterized in that pressed in step S3
According to 8-9log cfu/mL artificial contamination pathogenic bacteria.
3. a kind of method for collecting microorganism in fresh-cut fruit and vegetable according to claim 1 or 2, which is characterized in that step S4
Peptone water be 85-95mL 0.1wt%.
4. a kind of method for collecting microorganism in fresh-cut fruit and vegetable according to claim 3, which is characterized in that in step S5
Device includes: vegetable homogeneous collection (1), large aperture filter membrane (2), bacterium solution collector (3), small-bore filter membrane (4) and filter
Liquid displacer (5), which is characterized in that fruits and vegetables homogeneous collection (1), large aperture filter membrane (2), bacterium solution collector (3), aperture
Diameter filter membrane (4) and filtrate displacer (5) are according to being from top to bottom sequentially arranged.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910086839.4A CN109666590A (en) | 2019-01-29 | 2019-01-29 | A kind of method of microorganism in collection fresh-cut fruit and vegetable |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201910086839.4A CN109666590A (en) | 2019-01-29 | 2019-01-29 | A kind of method of microorganism in collection fresh-cut fruit and vegetable |
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| CN109666590A true CN109666590A (en) | 2019-04-23 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113999888A (en) * | 2021-12-16 | 2022-02-01 | 上海市农业科学院 | Method for eluting food-borne pathogenic bacteria |
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Patent Citations (3)
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| CN204911002U (en) * | 2015-09-15 | 2015-12-30 | 湖南赛隆药业有限公司 | Aseptic filteration device of preparation brain protein hydrolysis thing preparation |
| CN106811425A (en) * | 2015-11-27 | 2017-06-09 | 重庆北芝堂生物科技有限公司 | A kind of method that gut flora is separated from human excrement and urine |
| CN108896694A (en) * | 2018-07-05 | 2018-11-27 | 中国农业科学院农业质量标准与检测技术研究所 | A kind of remaining LC-QToF-MS Screening analysis method of pesticide in animal food |
Non-Patent Citations (1)
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| CN113999888A (en) * | 2021-12-16 | 2022-02-01 | 上海市农业科学院 | Method for eluting food-borne pathogenic bacteria |
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Application publication date: 20190423 |