CN106591416B - Chromogenic medium for detecting food-borne pathogenic yersinia - Google Patents
Chromogenic medium for detecting food-borne pathogenic yersinia Download PDFInfo
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- 241000607734 Yersinia <bacteria> Species 0.000 title claims abstract description 56
- 230000001717 pathogenic effect Effects 0.000 title claims abstract description 48
- 239000001963 growth medium Substances 0.000 claims abstract description 59
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 32
- 239000003833 bile salt Substances 0.000 claims abstract description 17
- 239000003593 chromogenic compound Substances 0.000 claims abstract description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 17
- 239000011780 sodium chloride Substances 0.000 claims abstract description 16
- 239000001888 Peptone Substances 0.000 claims abstract description 15
- 108010080698 Peptones Proteins 0.000 claims abstract description 15
- 235000019319 peptone Nutrition 0.000 claims abstract description 15
- PLXMOAALOJOTIY-FPTXNFDTSA-N Aesculin Natural products OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)[C@H]1Oc2cc3C=CC(=O)Oc3cc2O PLXMOAALOJOTIY-FPTXNFDTSA-N 0.000 claims abstract description 13
- WNBCMONIPIJTSB-BGNCJLHMSA-N Cichoriin Natural products O([C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1)c1c(O)cc2c(OC(=O)C=C2)c1 WNBCMONIPIJTSB-BGNCJLHMSA-N 0.000 claims abstract description 13
- XHCADAYNFIFUHF-TVKJYDDYSA-N esculin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC(C(=C1)O)=CC2=C1OC(=O)C=C2 XHCADAYNFIFUHF-TVKJYDDYSA-N 0.000 claims abstract description 13
- 229940093496 esculin Drugs 0.000 claims abstract description 13
- AWRMZKLXZLNBBK-UHFFFAOYSA-N esculin Natural products OC1OC(COc2cc3C=CC(=O)Oc3cc2O)C(O)C(O)C1O AWRMZKLXZLNBBK-UHFFFAOYSA-N 0.000 claims abstract description 13
- ZNCPFRVNHGOPAG-UHFFFAOYSA-L sodium oxalate Chemical compound [Na+].[Na+].[O-]C(=O)C([O-])=O ZNCPFRVNHGOPAG-UHFFFAOYSA-L 0.000 claims abstract description 13
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- FRHBOQMZUOWXQL-UHFFFAOYSA-L ammonium ferric citrate Chemical compound [NH4+].[Fe+3].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O FRHBOQMZUOWXQL-UHFFFAOYSA-L 0.000 claims abstract description 9
- 229960004642 ferric ammonium citrate Drugs 0.000 claims abstract description 9
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- 229960002950 novobiocin Drugs 0.000 claims description 7
- YJQPYGGHQPGBLI-KGSXXDOSSA-N novobiocin Chemical compound O1C(C)(C)[C@H](OC)[C@@H](OC(N)=O)[C@@H](O)[C@@H]1OC1=CC=C(C(O)=C(NC(=O)C=2C=C(CC=C(C)C)C(O)=CC=2)C(=O)O2)C2=C1C YJQPYGGHQPGBLI-KGSXXDOSSA-N 0.000 claims description 7
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- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 4
- 229930195725 Mannitol Natural products 0.000 description 4
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000588769 Proteus <enterobacteria> Species 0.000 description 2
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- 229910021641 deionized water Inorganic materials 0.000 description 2
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 2
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- 229940115939 shigella sonnei Drugs 0.000 description 2
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 2
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
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- 241000588748 Klebsiella Species 0.000 description 1
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- 241000588653 Neisseria Species 0.000 description 1
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 108010046334 Urease Proteins 0.000 description 1
- 241000607598 Vibrio Species 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
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- 102000005936 beta-Galactosidase Human genes 0.000 description 1
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- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical group CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 description 1
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- 159000000000 sodium salts Chemical class 0.000 description 1
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING 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
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/045—Culture media therefor
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Abstract
The invention discloses a chromogenic culture medium for detecting food-borne pathogenic yersinia, which contains soybean peptone, yeast powder, sodium chloride, sodium oxalate, agar, β -riboside chromogenic substrate, β -galactoside chromogenic substrate, esculin, ferric ammonium citrate, no-III bile salt, cephalosporin, neomycin and the balance of water.
Description
Technical Field
The invention relates to a culture medium for microbial detection, in particular to a chromogenic culture medium for detecting food-borne pathogenic yersinia, belonging to the field of microbial inspection and monitoring for food safety.
Background
The yersinia enterocolitica is one of important food-borne pathogenic bacteria, the food-borne pathogenic bacteria which can survive and proliferate in low-temperature stored food can cause extremely serious food poisoning events, the damage degree of the yersinia enterocolitica in individual countries even exceeds shigella, the yersinia enterocolitica is close to salmonella and campylobacter jejuni, once infection causes various diseases, the health of people is seriously threatened, the yersinia enterocolitica is a conventional inspection project for guaranteeing food safety in various countries, and the inspection method for the yersinia enterocolitica in the food is specified by the national food safety standard GB 4789.8.
Yersinia enterocolitica belongs taxonomically to the genus yersinia, of which there are 11 species, and it has been confirmed that yersinia pseudotuberculosis and yersinia pestis, which although causing cellulitis, are not transmitted through food, are pathogenic to humans, in addition to yersinia enterocolitica, which is pathogenic to humans, and thus, yersinia enterocolitica and yersinia pseudotuberculosis are referred to herein as yersinia enterocolitica and yersinia pseudotuberculosis.
For the detection method of yersinia enterocolitica in food, the conventional method is commonly used at present, taking the method of food safety standard GB4789.8 as an example, the specific operation is as follows: after the previous enrichment and the enrichment liquid treatment, the bacteria are inoculated to CIN-1 and an improved Y culture medium for culture and separation, then the suspected bacteria are subjected to a Klebsiella ferric test, a urease test, a semi-solid dynamic test and microscopic examination to obtain the result of whether the suspected bacteria belong to the Yersinia, and the Yersinia enterocolitica can be distinguished through the subsequent biochemical reaction, at least 6-7 days are needed, and the efficiency is low. In addition, methods such as a PCR technology and the like are also used for detecting the yersinia enterocolitica and other yersinia genus, special equipment and professional technicians are needed, the cost is high, the popularization and the application are not facilitated, and no special method for the food-borne pathogenic bacterium, namely the yersinia pseudotuberculosis exists at present.
The bacteria biochemical identification technology which is based on the traditional culture medium and is added with specific enzyme substrate can better meet the traditional method at present, the culture, the separation and the identification are completed at one time, the operation is simple and convenient, the detection time is greatly shortened, and the accuracy is improved through the characteristic identification of target bacteria, so the method has wide application prospect, is applied to the detection of bacteria such as salmonella, vibrio, listeria, pathogenic escherichia coli O157 and the like in a plurality of fields such as clinical medicine, food sanitation, environmental protection and the like, in the aspect of the application of the yersinia enterocolitica detection, the color development culture medium (CHROMAGARY. enterocolitica) is developed by the Cola corporation, but the culture medium mainly aims at the medical clinical examination and takes the highly pathogenic yersinia enterocolitica as a target, namely, the yersinia enterocolitica is divided into 6 groups according to phenotypic characteristics, and the color development culture medium developed by the Cola corporation can detect the residual 5 highly pathogenic yersinia enterocolitica with the highly pathogenic enterocolitica The bacteria (1B, 2,3,4 and 5 groups), namely the target bacteria which can be separated and identified by the culture medium, are not all the yersinia enterocolitica, and the yersinia pseudotuberculosis can not be separated and detected. The food safety test aims at reducing risks, and for yersinia enterocolitica, the method of the food safety standard GB4789.8 is taken as an example, and the clear test range is the whole biological group, so that the product cannot meet the requirements of food safety test, and in addition, the expensive price increases the cost of the use department in terms of application to medical test.
Therefore, there is a need in the art for a specific, efficient, rapid, inexpensive, and easy-to-use chromogenic medium.
Disclosure of Invention
The invention aims to provide a chromogenic culture medium which has high selectivity, short detection period, low cost and strong operability, is suitable for detecting yersinia in a food detection sample, and can distinguish yersinia enterocolitica and yersinia pseudotuberculosis which are main pathogenic yersinia while separating.
The culture medium is developed by screening and optimizing different factors such as carbon source, nitrogen source, inorganic salts, vitamins, pH value and the like by applying principles of biochemistry, microbiology and inorganic, organic and analytical chemistry.
The technical scheme adopted by the invention is as follows:
a chromogenic culture medium for separating and detecting food-borne pathogenic yersinia comprises, per 1000mL of culture medium, 15-20 g of peptone, 2-5 g of yeast powder, 2-5 g of sodium chloride, 1-2 g of sodium oxalate, 2-5 g of bile salt, 13-15 g of agar, 0-0.5 g of esculin, 0-0.5 g of ferric ammonium citrate, 0.05-0.08 g of β -riboside chromogenic substrate, 0.05-0.08 g of β -galactoside chromogenic substrate, 0-15 mg of cephalosporin, 0-2.5 mg of novobiocin, the balance of water, and the pH value of 7.4 +/-0.2.
In some embodiments, preferably, the peptone is soy peptone.
In some embodiments, preferably, the bile salt is a third-size bile salt.
In some embodiments, preferably, the β -riboside chromogenic substrate is 5-Bromo-4-chloro-3-indole- β -D-ribofuranoside (5-Bromo-4-chloro-3-indolyl- β -D-ribofuranoside).
In some embodiments, preferably, the β -galactoside chromogenic substrate is 5-Bromo-6-chloro-3-indole- β -D-galactopyranoside (5-Bromo-6-chloro-3-indolyl- β -D-galactopyranoside).
In some embodiments, preferably, each 1000mL of the culture medium comprises 15g of soybean peptone, 5g of yeast powder, 5g of sodium chloride, 2g of sodium oxalate, 2g of bile salts III, 13g of agar, 0.05g of 5 bromo-4-chloro-3-indole- β -D-ribofuranoside, 0.05g of 5 bromo-6-chloro-3-indole- β -D-galactopyranoside, and the balance of water, and the pH is 7.4 +/-0.2.
In some embodiments, preferably, each 1000mL of the culture medium comprises 15g of soybean peptone, 5g of yeast powder, 5g of sodium chloride, 2g of sodium oxalate, 2g of tribasic bile salt, 13g of agar, 0.05g of 5 bromo-4-chloro-3-indole- β -D-ribofuranoside, 0.05g of 5 bromo-6-chloro-3-indole- β -D-galactopyranoside, 15mg of cephalosporin, 2.5mg of novobiocin, and the balance water, and the pH is 7.4 +/-0.2.
In some embodiments, preferably, each 1000mL of the culture medium comprises 15g of soybean peptone, 5g of yeast powder, 5g of sodium chloride, 2g of sodium oxalate, 2g of bile salts III, 13g of agar, 0.5g of esculin, 0.5g of ferric ammonium citrate, 0.05g of 5 bromo-4-chloro-3-indole- β -D-ribofuranoside, 0.05g of 5 bromo-6-chloro-3-indole- β -D-galactopyranoside, 15mg of cephalosporin, 2.5mg of novobiocin, and the balance of water, and the pH is 7.4 +/-0.2.
According to the invention, bile salt and sodium oxalate are added into the culture medium, so that the growth of most gram-positive bacteria in a sample can be inhibited, and intestinal bacteria, especially proteus and pseudomonas are also mostly inhibited.
The invention adds antibiotics into the culture medium, the antibiotics can inhibit the growth of certain bacteria, and the preferable cephalosporin and the neomycin are broad-spectrum antibiotics which can inhibit the growth of gram-positive bacteria, dysentery bacillus, Neisseria, salmonella, proteus and other bacteria.
According to the invention, β -riboside chromogenic substrate 5-bromo-4-chloro-3-indole- β -D-ribofuranoside is added into a culture medium, so that bacteria capable of expressing β -ribosidase can decompose the chromogenic substrate under the action of enzyme to form a blue-green colony.
According to the invention, β -galactoside chromogenic substrate 5 bromo-6-chloro-3-indole- β -D-galactopyranoside is added into the culture medium, so that bacteria capable of expressing and producing β -galactosidase can decompose the chromogenic substrate under the action of enzyme to form purple colonies.
According to the invention, esculin and ferric ammonium citrate are added into the culture medium, esculin reacts positively, and turns into black around colonies, so that the culture medium is obviously contrasted with colonies without esculin decomposition. Of the 6 colonies of Yersinia enterocolitica, only colony 1A decomposed esculin to turn the colony periphery black. It is clearly indicated in Bergey handbook of bacteriological taxonomic Classification of enterocolitis Yersinia, group 1A, is a conditional pathogen or a weak pathogen, and groups 1B,2,3,4,5 are strongly pathogenic strains.
The method for separating and detecting food-borne pathogenic yersinia by using the chromogenic culture medium comprises the following steps:
1) preparation of color development plate: adding the components of the chromogenic culture medium except the antibiotics into deionized water, stirring for dissolving, adjusting the pH to 7.4 +/-0.2, heating for boiling, cooling to 45-55 ℃, adding the antibiotics, mixing uniformly, and pouring into a flat plate for later use;
2) sample treatment: processing according to the sample processing method specified by the corresponding standard specification. GB/T4789.8-2008 stipulates that yersinia enterocolitica in food is firstly subjected to improved phosphate buffer solution at 26 +/-1 ℃ and then subjected to enrichment for 48-72 hours, and then enriched liquid of other food except milk and dairy products is subjected to alkali treatment and fully mixed for 15 seconds.
3) Inoculating and culturing: inoculating the enrichment fluid of milk and dairy products or other food enrichment fluids after alkali treatment on a color development plate, and culturing for 24-36 h at 26 +/-1 ℃.
4) And (4) analyzing results: the purple colonies appeared to be suspected pathogenic yersinia.
The invention has the beneficial effects that:
the chromogenic medium can be used for separating and detecting food-borne pathogenic yersinia, has the advantages of strong specificity, high sensitivity, easiness in operation, simplicity in result judgment and the like, is suitable for detecting various samples, and has wide application prospects.
The chromogenic medium is divided into three chromogenic media according to different detection requirements, wherein one chromogenic medium is used for detecting food-borne pathogenic yersinia including yersinia pseudotuberculosis; the other is a chromogenic medium for detecting yersinia enterocolitica; the last one is a chromogenic medium for the detection of the strongly pathogenic yersinia enterocolitica.
Detailed Description
Example 1: specific experiment of yersinia color development medium of the invention on yersinia
1) Preparing a solid plate:
according to the formula, each 1000mL of culture medium comprises 15g of soybean peptone, 5g of yeast powder, 5g of sodium chloride, 2g of sodium oxalate, 2g of bile salt III, 13g of agar, 0.05g of 5 bromo-4-chloro-3-indole- β -D-ribofuranoside, 0.05g of 5 bromo-6-chloro-3-indole- β -D-galactopyranoside and 0.05g of water in balance, wherein the pH is 7.4 +/-0.2;
2) strain activation and culture:
the test bacteria include Staphylococcus aureus ATCC25923, Escherichia coli IQCC10113 producing enterotoxigenia, Enterobacter sakazakii ATCC29544, Salmonella typhimurium ATCC14028, Aeromonas veterinaria ATCC51108, Yersinia intermedia ATCC33647, Yersinia Aromatica ATCC35236, Yersinia ke ATCC33639, Yersinia pseudotuberculosis CMCC53504, Yersinia enterocolitica standard strain ATCC9610, ATCC23715 and isolates IQCC10901, IQCC10903, IQCC10904 and IQCC 10906. ATCC, shorthand for American type culture Collection; IQCC is a shorthand for the China Inspection and Quarantine microorganism Culture Collection (Inspection and Quarantine Culture Collection).
Respectively activating the above strains, inoculating to Columbia blood agar plate, and optimizingCulturing at the temperature for 18-24 h until obvious bacterial colonies appear, respectively selecting 3-5 bacterial colonies, dispersing in a TSB liquid culture medium to prepare bacterial suspension, adjusting the turbidity of the bacterial suspension to 0.5 McLeod's turbidimetric standard, and then the concentration of the bacterial suspension is 1.5 × 108CFU/mL, use.
3) Inoculation: and (3) respectively taking a ring of streaks from bacterial suspensions prepared from the 15 strains, inoculating the streaks on a prepared color development plate, culturing all the bacteria at 26 +/-1 ℃ for 24-36 h to avoid the influence of temperature, and observing the color development condition of bacterial colonies.
4) And (4) analyzing results: the food-borne pathogenic yersinia grows well on the flat plate, the center part is purple, the edge is semitransparent, and the diameter of a bacterial colony is 1-3 mm; the non-target bacteria are inhibited or present in colonies of different colors, and the colony morphology of other yersinia is similar to that of the pathogenic yersinia, and the detailed results are shown in table 1 and the label.
Table 1: experimental results of example 1
| Test strains | Growth conditions | Color of colony |
| Staphylococcus aureus ATCC25923 | - | Suppression of |
| Enterotoxigenic escherichia coli IQCC10113 | +++ | Blue green color |
| Enterobacter sakazakii ATCC29544 | +++ | Tan colora |
| Salmonella typhimurium ATCC14028 | +++ | Blue green color |
| Aeromonas veterinarian ATCC51108 | +++ | Blue green color |
| Yersinia intermedia ATCC33647 | +++ | Purple color |
| Yersinia aryabhattai ATCC35236 | +++ | Purple colorb |
| Yersinia kloniae ATCC33639 | +++ | Bluish violetc |
| Yersinia pseudotuberculosis CMCC53504 | ++ | Purple colord |
| Yersinia enterocolitica ATCC9610 | +++ | Purple color |
| Yersinia enterocolitica ATCC23715 | +++ | Purple color |
| Yersinia enterocolitica IQCC10901 | +++ | Purple color |
| Yersinia enterocolitica IQCC10903 | +++ | Purple color |
| Yersinia enterocolitica IQCC10904 | +++ | Purple color |
| Yersinia enterocolitica IQCC10906 | +++ | Purple color |
a, Enterobacter sakazakii produces yellow pigment in the culture medium and is mixed with a chromogenic substrate to present a tan colony;
b, when the yersinia alismoides grows in the culture medium for 24 hours, the colony has no color or only a small part in the center turns purple, and the transparent ring at the edge of the colony is obvious.
And c, when the yersinia clarkii grows in the culture medium for 24 hours, the colony presents blue-green color, and when the culture is continued for 36 hours, the colony gradually changes into blue-purple color.
Yersinia pseudotuberculosis grows relatively slowly in the culture medium, and the colony edges are irregular in shape.
The experimental result shows that the culture medium can well distinguish food-borne pathogenic yersinia and other non-target bacteria.
Example 2: selective effect experiment of yersinia color development culture medium
1) Preparing a solid plate:
according to the formula, each 1000mL of culture medium comprises 15g of soybean peptone, 5g of yeast powder, 5g of sodium chloride, 2g of sodium oxalate, 2g of bile salt III, 13g of agar, 0.05g of 5 bromo-4-chloro-3-indole- β -D-ribofuranoside, 0.05g of 5 bromo-6-chloro-3-indole- β -D-galactopyranoside, 15mg of cephalosporin, 2.5mg of novobiocin and the balance of water, wherein the pH is 7.4 +/-0.2;
2) strain activation and culture:
the standard strain tested is described in example 1.
Respectively activating the strains, inoculating the activated strains to a Columbia blood agar plate, culturing for 18-24 h at a corresponding optimal temperature until obvious bacterial colonies appear, respectively selecting 3-5 bacterial colonies, dispersing in a TSB liquid culture medium to prepare bacterial suspension, adjusting the turbidity of the bacterial suspension to 0.5 McLeeb turbidimetric standard, and adjusting the concentration of the bacterial suspension to 1.5 × 108CFU/mL, use.
3) Inoculation: and (3) respectively taking a ring of streaks from bacterial suspensions prepared from the 15 strains, inoculating the streaks on a prepared color development plate, culturing all the bacteria at 26 +/-1 ℃ for 24-36 h to avoid the influence of temperature, and observing the color development condition of bacterial colonies.
4) And (4) analyzing results: after addition of antibiotics, both Salmonella and Yersinia pseudotuberculosis were inhibited. The yersinia enterocolitica grows well on the flat plate, the center part is purple, the edge is semitransparent, and the diameter of a bacterial colony is 1-3 mm; other non-target bacteria are inhibited from growing or the colonies are in other colors. The detailed results are shown in Table 2 and noted.
Table 2: experimental results of example 2
| Test strains | Growth conditions | Color of colony |
| Staphylococcus aureus ATCC25923 | - | Suppression of |
| Enterotoxigenic escherichia coli IQCC10113 | +++ | Blue green color |
| Enterobacter sakazakii ATCC29544 | +++ | Tan colora |
| Salmonella typhimurium ATCC14028 | - | Suppression of |
| Aeromonas veterinarian ATCC51108 | +++ | Blue green color |
| Yersinia intermedia ATCC33647 | +++ | Purple color |
| Yersinia aryabhattai ATCC35236 | +++ | Purple colorb |
| Yersinia kloniae ATCC33639 | +++ | Bluish violetc |
| Yersinia pseudotuberculosis CMCC53504 | - | Suppression of |
| Yersinia enterocolitica ATCC9610 | ++ | Purple color |
| Yersinia enterocolitica ATCC23715 | +++ | Purple color |
| Yersinia enterocolitica IQCC10901 | +++ | Purple color |
| Yersinia enterocolitica IQCC10903 | +++ | Purple color |
| Yersinia enterocolitica IQCC10904 | +++ | Purple color |
| Yersinia enterocolitica IQCC10906 | +++ | Purple color |
a, Enterobacter sakazakii produces yellow pigment in the culture medium and is mixed with a chromogenic substrate to present a tan colony;
b, when the yersinia alismoides grows in the culture medium for 24 hours, the colony has no color or only a small part in the center turns purple, and the transparent ring at the edge of the colony is obvious.
And c, when the yersinia clarkii grows in the culture medium for 24 hours, the colony presents blue-green color, and when the culture is continued for 36 hours, the colony gradually changes into blue-purple color.
The experimental result shows that the culture medium can well distinguish the enterocolitis yersinia and other non-target bacteria. After the corresponding antibiotics are added, both salmonella and yersinia pseudotuberculosis are inhibited and cannot grow in the culture medium, and the growth of the yersinia enterocolitica ATCC9610 is also affected by the corresponding antibiotics, is weaker than that of other yersinia enterocolitica and can be obviously distinguished from other non-target bacteria. Some species of partially non-pathogenic yersinia, such as yersinia intermedia, may also form a purplish colony similar to the pathogenic yersinia, and suspected bacteria may be directly distinguished by additional experiments, such as API 20E.
Example 3: differentiation of yersinia genus chromogenic medium of the present invention on highly pathogenic enterocolitis yersinia genus
1) Preparing a solid plate:
according to the formula, each 1000mL of culture medium comprises 15g of soybean peptone, 5g of yeast powder, 5g of sodium chloride, 2g of sodium oxalate, 2g of bile salt III, 13g of agar, 0.5g of esculin, 0.5g of ferric ammonium citrate, 0.05g of 5 bromo-4-chloro-3-indole- β -D-ribofuranoside, 0.05g of 5 bromo-6-chloro-3-indole- β -D-galactopyranoside, 15mg of cephalosporin, 2.5mg of neomycin and the balance of water, wherein the pH is 7.4 +/-0.2, the culture medium is prepared by adding the components of the chromogenic culture medium except the antibiotics into deionized water, stirring and dissolving, adjusting the pH to 7.4 +/-0.2, heating and boiling, cooling to 45-55 ℃, adding the corresponding antibiotics, performing aseptic operation, and pouring a plate for later use;
2) strain activation and culture:
the tested bacteria comprise staphylococcus aureus ATCC25923, Shigella sonnei IQCC11320, Enterobacter sakazakii ATCC29544, Salmonella typhimurium ATCC14028, Aeromonas veterinaria ATCC51108, Yersinia intermedia ATCC33647, Yersinia arrhizus ATCC35236, Yersinia kirschner ATCC33639, Yersinia pseudotuberculosis CMCC53504, Yersinia enterocolitica standard strain ATCC9610, ATCC23715 and isolates IQCC10901, IQCC10903, IQCC10904 and IQCC 10906.
Respectively activating the strains, inoculating the activated strains to a Columbia blood agar plate, culturing for 18-24 h at a corresponding optimal temperature until obvious colonies appear, respectively selecting 3-5 colonies, dispersing in a TSB liquid culture medium to prepare a bacterial suspension, adjusting the turbidity of the bacterial suspension to 0.5 McBt standard, and using the bacterial suspension with the concentration of 1.5 × 108 CFU/mL.
3) Inoculation: and (3) respectively taking a ring of streaks from bacterial suspensions prepared from the 15 strains, inoculating the streaks on a prepared color development plate, culturing all the bacteria at 26 +/-1 ℃ for 24-36 h to avoid the influence of temperature, and observing the color development condition of bacterial colonies.
4) And (4) analyzing results: according to previous studies, yersinia enterocolitica was divided into 6 groups, and only group 1A decomposed esculin to turn the colony black. It is specifically stated in Bojie's handbook of bacteriological taxonomic Classification of bacteria that Yersinia enterocolitica group 1A is a conditional pathogen or a weak pathogen, and groups 1B,2,3,4, and 5 are strongly pathogenic strains. Therefore, after esculin and ferric ammonium citrate are added into the culture medium, whether the isolated yersinia enterocolitica is a strongly pathogenic strain can be judged according to whether the periphery of the colony becomes black or not. In this medium, gram-positive bacteria, Salmonella, Shigella and Yersinia pseudotuberculosis were all completely inhibited. The detailed results are shown in Table 3.
Table 3: experimental results of example 3
| Test strains | Growth conditions | Color of colony |
| Staphylococcus aureus ATCC25923 | - | Suppression of |
| Shigella sonnei IQCC11320 | - | Suppression of |
| Enterobacter sakazakii ATCC29544 | +++ | Tan coloraBlackening the surroundings |
| Salmonella typhimurium ATCC14028 | - | Suppression of |
| Aeromonas veterinarian ATCC51108 | +++ | Bluish green, blackened surroundings |
| Yersinia intermedia ATCC33647 | +++ | Purple color and blackened surroundings |
| Yersinia aryabhattai ATCC35236 | +++ | Purple colorb |
| Yersinia kloniae ATCC33639 | +++ | Bluish violetc |
| Yersinia pseudotuberculosis CMCC53504 | - | Suppression of |
| Yersinia enterocolitica ATCC9610 | ++ | Purple color |
| Yersinia enterocolitica ATCC23715 | +++ | Purple color |
| Yersinia enterocolitica IQCC10901 | +++ | Purple color and blackened surroundings |
| Yersinia enterocolitica IQCC10903 | +++ | Purple color and blackened surroundings |
| Yersinia enterocolitica IQCC10904 | +++ | Purple color and blackened surroundings |
| Yersinia enterocolitica IQCC10906 | +++ | Purple color and blackened surroundings |
a, Enterobacter sakazakii produces yellow pigment in the culture medium and is mixed with a chromogenic substrate to present a tan colony;
b, when the yersinia alismoides grows in the culture medium for 24 hours, the colony has no color or only the central part turns purple, and the transparent ring at the edge of the colony is obvious.
And c, when the yersinia clarkii grows in the culture medium for 24 hours, the colony presents blue-green color, and when the culture is continued for 36 hours, the colony gradually changes into blue-purple color.
The experimental result shows that the culture medium not only can well distinguish the enterocolitis yersinia from other non-target bacteria, but also can distinguish strong pathogenic strains and conditional pathogenic strains in the enterocolitis yersinia. After addition of esculin and ferric ammonium citrate, yersinia intermedia can also be easily distinguished from yersinia enterocolitica, which is a highly pathogenic bacterium.
The culture medium can be applied to the separation and detection of the highly pathogenic yersinia enterocolitica, the growth on the culture medium is vigorous, bacterial colonies are purple, transparent bacteria rings exist around the bacterial colonies, and the bacterial colonies which are not discolored around the bacterial colonies are the highly pathogenic yersinia enterocolitica.
Example 4: sample pretreatment method using yersinia color development culture medium
The sample pretreatment method comprises the following steps: pre-culturing and alkali treating with improved phosphate buffer solution;
1) preparing a related culture medium:
preparing a solid plate: reference example 1;
preparation of modified phosphate buffer (mannitol): each 1000mL of the culture medium contained 8.23g of disodium hydrogen phosphate, 1.2g of sodium dihydrogen phosphate, 5g of sodium chloride, 1.5g of tribasic sodium salt, 20g of mannitol and the balance of water, and the pH was adjusted to 7.6. Sterilizing at 121 deg.C under high pressure for 15min, and packaging.
Preparing an alkali treatment solution: the 0.5% sodium chloride solution and the 0.5% potassium hydroxide solution were mixed in equal amounts.
2) Preparing an artificial pollution sample: samples (commercial pork) were inoculated at two levels of inoculation (10)2CFU/25g and 105CFU/25 g), adding Yersinia enterocolitica ATCC23715, homogenizing, mixing, and pre-culturing.
3) Sample treatment: adding 225ml of improved phosphate buffer solution into two inoculated level artificially contaminated samples, enriching for 48 hours at 26 +/-1 ℃, then respectively sucking 0.5ml of enriched liquid, adding 4.5ml of alkali treatment liquid, fully mixing for 15s after alkali treatment, and adding 4.5ml of sterile water into a control group.
4) Inoculating and culturing: respectively inoculating 4 tubes of enrichment liquid on solid plates, coating 3 plates on each tube of enrichment liquid, and culturing at 26 +/-1 ℃ for 24-36 h.
5) And (4) analyzing results: most of other bacteria which are not alkali-resistant can be killed by using alkali treatment, the detection rate is improved, and detailed results are shown in table 4. Therefore, when the medium of the present invention is used to screen a target bacterium, it is recommended to perform streaking culture after alkali treatment.
Table 4: influence of alkali treatment on detection rate of target bacteria in sample
Example 5 Yersinia pseudotuberculosis in Yersinia chromogenic Medium according to the invention isolation of Yersinia pseudotuberculosis from samples
1) Preparing a related culture medium:
preparing a solid plate: reference example 1;
preparation of modified phosphate buffer (mannitol) and alkaline treatment solution: reference example 4;
2) preparing an artificial pollution sample: samples (commercial pork) were run at 102The inoculation level of CFU/25g is inoculated with Yersinia pseudotuberculosis CMCC53504, and the mixture is homogenized and mixed for pre-culture.
3) Sample treatment: adding the artificially contaminated sample into 225ml of modified phosphate buffer (mannitol), enriching bacteria for 48h at 26 +/-1 ℃, sucking 0.5ml of enriched bacteria liquid, adding 4.5ml of alkali treatment liquid, and fully mixing for 15s after alkali treatment.
4) Inoculating and culturing: inoculating the enrichment broth onto a solid plate, and culturing at 26 +/-1 ℃ for 24-36 h.
5) And (4) analyzing results: yersinia pseudotuberculosis CMCC53504 grows vigorously on this medium, and the colonies are purple. Yersinia pseudotuberculosis can be isolated and detected from this medium.
Example 6 Yersinia enterocolitica isolation from samples by Yersinia chromogenic Medium according to the invention
1) Preparing a related culture medium:
preparing a solid plate: performing sterile operation according to example 2 or example 3, and pouring the plate for standby;
preparation of modified phosphate buffer (sorbitol): each 1000mL of the culture medium contained 8.23g of disodium hydrogen phosphate, 1.2g of sodium dihydrogen phosphate, 5g of sodium chloride, 1.5g of tribasic sodium chloride, 20g of sorbitol, and the balance of water, and the pH was adjusted to 7.6. Sterilizing at 121 deg.C under high pressure for 15min, and packaging.
Preparing an alkali treatment solution: reference example 4;
2) preparing an artificial pollution sample: samples (commercial pork filling) were run at 102The inoculation level of CFU/25g is respectively inoculated with a strong pathogenic strain ATCC9610 and a weak pathogenic strain IQCC10906 of the yersinia enterocolitica, and the mixture is homogenized and mixed respectively for pre-culture.
3) Sample treatment: respectively adding the two artificial pollution samples into 225ml of improved phosphate buffer solution (sorbitol), after increasing bacteria for 48 hours at 26 +/-1 ℃, respectively sucking 0.5ml of bacteria increasing solution, adding 4.5ml of alkali treatment solution, and fully mixing for 15s after alkali treatment.
4) Inoculating and culturing: and respectively inoculating the two enrichment fluids to the solid plates prepared in the formulas of the embodiment 2 and the embodiment 3, and culturing at 26 +/-1 ℃ for 24-36 h.
5) And (4) analyzing results: both the example 2 and example 3 media can be applied to the isolation and detection of yersinia enterocolitica, and the example 3 media can be used to distinguish between highly pathogenic and conditionally pathogenic yersinia enterocolitica. Yersinia enterocolitica grows vigorously on the medium, colonies are purple, and clear fungus rings exist around the colonies. The colonies surrounding the colonies were not discolored by the strongly pathogenic yersinia enterocolitica, whereas the conditionally pathogenic yersinia enterocolitica. The user can select the culture medium with the corresponding formula according to specific requirements. The detailed results are shown in Table 5.
Table 5: experimental results of example 6
The foregoing detailed description of the embodiments is illustrative rather than limiting in nature and that several embodiments are presented in the limited sense, such that variations and modifications are possible without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (8)
1. The chromogenic culture medium for separating and detecting food-borne pathogenic yersinia is characterized in that every 1000mL of the culture medium comprises 15-20 g of peptone, 2-5 g of yeast powder, 2-5 g of sodium chloride, 1-2 g of sodium oxalate, 2-5 g of bile salt, 13-15 g of agar, 0-0.5 g of esculin, 0-0.5 g of ferric ammonium citrate, 0.05-0.08 g of β -riboside chromogenic substrate, 0.05-0.08 g of β -galactoside chromogenic substrate, 0-15 mg of cephalosporin, 0-2.5 mg of novobiocin, the balance of water, and the pH value is 7.4 +/-0.2.
2. The chromogenic medium for the isolation and detection of food-borne pathogenic yersinia according to claim 1, wherein said peptone is tryptone, soytone, bacterial peptone or a mixture thereof.
3. The chromogenic medium for isolating and detecting food-borne pathogenic yersinia according to claim 1, wherein the bile salt is at least one of sodium deoxycholate, porcine bile salt, mixed bile salt, bovine bile salt and third bile salt.
4. The chromogenic medium for isolating and detecting yersinia pathogenic for food origin according to claim 1, wherein the β -riboside chromogenic substrate is 5 Bromo-4-chloro-3-indole- β -D-ribofuranoside (5-Bromo-4-chloro-3-indolyl- β -D-ribofuranoside).
5. The chromogenic medium for isolating and detecting yersinia pathogenic for food origin according to claim 1, wherein β -galactoside chromogenic substrate is 5 Bromo-6-chloro-3-indole- β -D-galactopyranoside (5-Bromo-6-chloro-3-indolyl- β -D-galactopyranoside).
6. The chromogenic medium for isolating and detecting food-borne pathogenic Yersinia according to claim 1, wherein each 1000mL of the medium comprises soybean peptone 15g, yeast powder 5g, sodium chloride 5g, sodium oxalate 2g, No. 2g, agar 13g, 5-bromo-4-chloro-3-indole- β -D-ribofuranoside 0.05g, 5-bromo-6-chloro-3-indole- β -D-galactopyranoside 0.05g, and the balance water, and has a pH of 7.4. + -. 0.2.
7. The chromogenic medium for isolating and detecting food-borne pathogenic Yersinia according to claim 1, wherein each 1000mL of the medium comprises soybean peptone 15g, yeast powder 5g, sodium chloride 5g, sodium oxalate 2g, No. 2g, agar 13g, 5-bromo-4-chloro-3-indole- β -D-ribofuranoside 0.05g, 5-bromo-6-chloro-3-indole- β -D-galactopyranoside 0.05g, cephalosporin 15mg, novobiocin 2.5mg, balance water, pH 7.4. + -. 0.2.
8. The chromogenic medium for isolating and detecting food-borne pathogenic Yersinia according to claim 1, wherein each 1000mL of the medium comprises soybean peptone 15g, yeast powder 5g, sodium chloride 5g, sodium oxalate 2g, No. 2g, agar 13g, esculin 0.5g, ferric ammonium citrate 0.5g, 5-bromo-4-chloro-3-indole- β -D-ribofuranoside 0.05g, 5-bromo-6-chloro-3-indole- β -D-galactopyranoside 0.05g, cephalosporin 15mg, novobiocin 2.5mg, balance water, and pH 7.4. + -. 0.2.
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| CN108374037B (en) * | 2018-05-14 | 2021-09-07 | 广东达元绿洲食品安全科技股份有限公司 | Chromogenic medium for yersinia enterocolitica and application of chromogenic medium |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0881284A1 (en) * | 1997-05-13 | 1998-12-02 | Société La Biochimie Appliquée - SO.LA.BI.A. | Culture medium for the specific detection of enterobacteries, procedure for its obtention and use |
| CN102816715A (en) * | 2012-08-01 | 2012-12-12 | 上海市疾病预防控制中心 | Kit for separating and identifying yersinia enterocolitica as well as preparation and application thereof |
-
2016
- 2016-10-27 CN CN201610954813.3A patent/CN106591416B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0881284A1 (en) * | 1997-05-13 | 1998-12-02 | Société La Biochimie Appliquée - SO.LA.BI.A. | Culture medium for the specific detection of enterobacteries, procedure for its obtention and use |
| CN102816715A (en) * | 2012-08-01 | 2012-12-12 | 上海市疾病预防控制中心 | Kit for separating and identifying yersinia enterocolitica as well as preparation and application thereof |
Non-Patent Citations (6)
| Title |
|---|
| Yersinia pestis lacZ expresses a b-galactosidasewith lowenzymatic activity;Alexander G. Bobrov等;《FEMS Microbiol Lett》;20060131;43–51 * |
| 两种培养基对病原性耶尔森氏菌检测的效果观察;曹淑兰等;《中国地方病防治杂志》;20030630;152-153 * |
| 几种耶氏菌分离琼脂培养基的比较;潘天太等;《中国人兽共患病杂志》;19861227;54 * |
| 小肠结肠炎耶尔森氏菌的增菌培养基;杜景红等;《中国畜禽传染病》;19940430;48-49 * |
| 小肠结肠炎耶尔森氏菌自凝试验培养基的研究;陈亭平;《广西医科大学学报》;19951230;489-493 * |
| 耶尔森氏菌感染与培养基研究;辜明铭等;《国外医学临床生物化学与检验学分册》;19910630;106-110 * |
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Effective date of registration: 20210611 Address after: Room 409-87, 4th floor, building 1, 38 Yongda Road, Daxing biomedical industrial base, Zhongguancun Science and Technology Park, Daxing District, Beijing 102600 (cluster registration) Patentee after: BEIJING JUNLIKANG BIOTECHNOLOGY Co.,Ltd. Address before: 100123 No. 3, Gaobeidian North Road, Beijing, Chaoyang District Patentee before: CHINESE ACADEMY OF INSPECTION AND QUARANTINE |
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