CN106755274B - Separation and identification method of swine escherichia coli - Google Patents
Separation and identification method of swine escherichia coli Download PDFInfo
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- CN106755274B CN106755274B CN201611074505.8A CN201611074505A CN106755274B CN 106755274 B CN106755274 B CN 106755274B CN 201611074505 A CN201611074505 A CN 201611074505A CN 106755274 B CN106755274 B CN 106755274B
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Abstract
The invention discloses a separation and identification method of swine escherichia coli, belonging to the technical field of microorganisms. The method mainly comprises three steps of bacteria isolation culture, gram staining, microscopic examination and biochemical identification test, and is simple, short in time consumption, low in cost and accurate and reliable in identification result. The bacteria isolation culture sequentially adopts a nutrient broth culture medium, a MacConkey agar culture medium and an eosin methylene blue agar culture medium for selective culture, and the grown bacterial colony has clear morphology and is convenient to pick and separate.
Description
Technical Field
The invention relates to a separation and identification method of swine escherichia coli, belonging to the technical field of microorganisms.
Background
Coli is a zoonosis pathogen that widely exists in nature and can cause human and animal co-infection, and can be classified into 3 types according to its pathogenic mechanism: symbiotic, enteropathogenic and extraintestinal pathogenic. Colibacillosis is an acute, polymorphic, enteric infectious disease caused mainly by the pathogenic Escherichia coli. In veterinary clinic, due to differences of serotype, immune state, physiological function, day age of pigs and the like of pathogenic escherichia coli, common pig colibacillosis is divided into 3 types, namely yellow scour of piglets with 3 days age, white scour of piglets with 7-10 days age and edema disease of piglets with 30 days age. Colibacillosis of pigs occurs all over the world, yellow and white scours of piglets are the most common infectious diseases in pig farms, the morbidity and mortality of the yellow and white scours of piglets are the first diseases of epidemic diseases of piglets, and great harm and serious economic loss are brought to the pig raising industry. After colibacillosis occurs, antibiotics are mostly used for treatment, but due to the wide and unreasonable use of antibiotics, the problem of bacterial drug resistance is increasingly serious, and particularly the problem of multiple drug resistance is more prominent. In order to prevent and cure the disease, various types of genetic engineering bacterins and multivalent fire-extinguishing bacterins are developed at home and abroad, but the immunity effect is not ideal due to the complex serotype of escherichia coli and the difference of epidemic strains in various regions, and the self-made fire-extinguishing bacterin effect is generally good.
In order to understand and master the drug resistance degree of escherichia coli in various regions and provide a basis for screening clinically effective prevention and treatment drugs, biochemical and serotype identification of pathogenic escherichia coli in collected samples is necessary. And the general identification procedure is: isolated culture → picking suspicious colonies → pure culture → gram stain, microscopic examination → biochemical test. The identification steps are as follows: diluting a disease material with normal saline, inoculating the disease material on a common culture medium, culturing at a constant temperature of 37 ℃ for 18-24H, observing the growth condition and the colony characteristics of bacteria, selecting a single colony to inoculate on a Macconkey agar plate, culturing at a constant temperature of 37 ℃ for 18-24H, then selecting red and medium-sized smooth colonies to perform gram staining and microscopic examination, primarily judging strains with staining conditions, shapes, sizes and the like conforming to the characteristics of escherichia coli as escherichia coli, selecting suspected strains to perform indole test, MR test, VP test, H test and the like2S test, ferric trisaccharide test and sugar fermentation test, and finally judging the strain genus by combining the biochemical test result. However, the identification result of the method is not accurate enough, and particularly, the colony morphology is fuzzy in the separation and purification operation, which is not beneficial to picking and separating.
Disclosure of Invention
The invention aims to provide a separation and identification method of swine escherichia coli, which adopts a nutrient broth culture medium, a Mackanka culture medium and an eosin methylene blue culture medium to separate and purify pathogenic strains, and has the advantages of clear growing bacterial colony morphology, convenience in picking and separating, simplicity and convenience in operation, accurate and reliable identification result and small error.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
a separation and identification method of Escherichia coli from swine comprises the following steps:
1) and (3) bacteria isolation culture: inoculating the collected sample into a nutrient broth culture medium, taking a bacterium solution after culture, inoculating the bacterium solution on a Macconkey agar culture medium in a streaking manner, observing the growth condition of bacterial colonies, selecting single or paired red bacterial colonies to inoculate on an eosin methylene blue agar culture medium, observing the growth condition of the bacterial colonies, and selecting single or paired purple black bacterial colonies for later use;
2) gram staining and microscopic examination;
3) biochemical identification test;
the nutrient broth culture medium in step 1) consists of: 7-8 g of peptone, 2-2.5 g of beef extract, 3.5-4 g of sodium chloride, 750mL of distilled water and 7.0-7.4 of pH value; the composition of the Mackanka agar medium is as follows: 12-18 g of peptone, 3.5-4 g of bile salt, 3.5-4 g of sodium chloride, 10-15 g of agar, 7-8 g of lactose, 0.01-0.03 g of neutral red, 750mL of distilled water and 7.0-7.4 of pH value; the composition of eosin methylene blue agar medium is: 7.5-8.5 g of peptone, 7-8 g of lactose, 1.2-1.8 g of dipotassium hydrogen phosphate, 15-20 g of agar, 12-18 mL of 2% eosin aqueous solution, 10-15 mL of 0.5% methylene blue aqueous solution, 750mL of distilled water and 7.2-7.4 of pH value.
Preferably, the composition of the nutrient broth in step 1) is: 7.5g of peptone, 2.3g of beef extract, 3.8g of sodium chloride, 750mL of distilled water and 7.2 of pH value; the composition of the Mackanka agar medium is as follows: 15g of peptone, 3.8g of bile salt, 3.8g of sodium chloride, 12g of agar, 7.5g of lactose, 0.02g of neutral red, 750mL of distilled water and 7.2 of pH value; the composition of eosin methylene blue agar medium is: 8g of peptone, 7.5g of lactose, 1.5g of dipotassium phosphate, 18g of agar, 15mL of 2% eosin aqueous solution, 12mL of 0.5% methylene blue aqueous solution, 750mL of distilled water and 7.3 of pH value.
The temperature for culturing in the step 1) is 36-38 ℃, and the culturing time is 15-20 h. Preferably, the culture is carried out at 37 ℃ for 18 h.
The gram staining in the step 2) mainly comprises four steps of primary staining, mordant staining, decoloring and counterstaining. The specific operation is as follows: fixing the selected colony smear, primarily dyeing the colony smear with ammonium oxalate crystal violet, mordanting the colony smear with iodine solution, and counterdyeing the colony smear with a re-reddening dye solution after decoloring.
The microscopic examination in the step 2) is as follows: covering the bacteria coating part with cedar oil, observing the dyeing condition and the bacterial morphology under a microscope, and selecting gram-negative bacteria for later use.
In the step 3), bacteria micro biochemical reaction tubes (such as Enterobacteriaceae bacteria biochemical coding identification tubes purchased from Hangzhou Tian and microbial reagents, Inc.) are adopted for biochemical identification. The specific operation is shown in the kit instruction.
And 3) performing strain preservation on the strain biochemically identified as the Escherichia coli in the step 3). The specific operation is as follows: inoculating the strain into nutrient broth culture medium, culturing, mixing the strain liquid with sterilized glycerol to obtain glycerol strain, and storing at a temperature below-20 deg.C.
The invention has the beneficial effects that:
the separation and identification of the swine escherichia coli mainly comprises three steps of bacteria separation and culture, gram staining, microscopic examination and biochemical identification test. The bacteria isolation culture sequentially adopts a nutrient broth culture medium, a MacConkey agar culture medium and an eosin methylene blue agar culture medium for selective culture, and the grown bacterial colony has clear morphology and is convenient to pick and separate.
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FIG. 1 is a morphological diagram of a colony in example 1;
FIG. 2 is a morphological diagram of bacteria examined under the microscope.
Detailed Description
The following examples are intended to illustrate the invention in further detail, but are not to be construed as limiting the invention in any way.
Example 1
The separation and identification method of the swine escherichia coli comprises the following steps:
1) collecting a sample: collecting 40 parts of excrement of an unweaned piglet and a diarrhea piglet from a large-scale pig farm in a certain county;
2) and (3) bacteria isolation culture: respectively inoculating collected pig manure samples into nutrient broth culture media, placing the nutrient broth culture media on a shaking table, culturing for 18h at 37 ℃ and 150rpm, picking a small amount of bacterial liquid by using an aseptic inoculating rod after culturing, streaking and inoculating the bacterial liquid on a Macconkey agar culture medium, culturing for 18h in a 37 ℃ culture box, observing the growth condition of bacterial colonies, picking pink, single or paired bacterial colonies, inoculating the bacterial colonies on an eosin Meilan agar culture medium, culturing for 18h in the 37 ℃ culture box, observing the growth condition of the bacterial colonies, picking purple black, single or paired bacterial colonies for later use, and co-separating 40 collected samples to obtain 17 strains;
the nutrient broth culture medium comprises the following components: 7.5g of peptone, 2.3g of beef extract, 3.8g of sodium chloride, 750mL of distilled water and 7.2 of pH value; the composition of the Mackanka agar medium is as follows: 15g of peptone, 3.8g of bile salt, 3.8g of sodium chloride, 12g of agar, 7.5g of lactose, 0.02g of neutral red, 750mL of distilled water and 7.2 of pH value; the composition of eosin methylene blue agar medium is: 8g of peptone, 7.5g of lactose, 1.5g of dipotassium phosphate, 18g of agar, 15mL of 2% eosin aqueous solution, 12mL of 0.5% methylene blue aqueous solution, 750mL of distilled water and 7.3 of pH value;
the results show that: the separated and cultured colonies all accord with the colony morphology of the escherichia coli (see figure 1, wherein A is the colony morphology on a MacconKa culture medium, and B is the colony morphology obtained on an eosin methylene blue culture medium);
3) gram staining: smearing and fixing picked bacterial colonies by flame, dripping 2 drops of ammonium oxalate crystal violet solution on a glass slide, dyeing for 1min, washing by using tap water, dripping 2 drops of iodine solution again, dyeing for 1min, washing by using tap water, blotting the glass slide by using absorbent paper, continuously dripping 95% ethanol solution for decoloring, washing after 30s, blotting the glass slide by using absorbent paper, finally dripping 2 drops of reddening dye solution, counterdyeing for 1min, and washing by using tap water;
microscopic examination: after dyeing, covering the bacterium coating part with cedar oil, placing the slide under a microscope for observation, finding out the bacterium with a low-power lens, and observing the dyeing condition and the bacterium shape with a high-power objective lens;
the dyeing and microscopic examination results show that: the 17 strains are gram-negative bacteria, and accord with the bacterial form of escherichia coli (shown in figure 2), and the two ends are blunt round, single or paired, and are in a shape of a tiny short rod;
4) biochemical identification test: the 17 strains of bacteria are identified by adopting a micro-identification tube (purchased from Hangzhou Tian and microbial agents Co., Ltd.) for biochemical coding of the bacteria of Enterobacteriaceae, and the specific operation is as follows: respectively breaking the bacteria biochemical identification tube from the middle of the identification tube by using a grinding wheel scratch, taking two ends of the identification tube as a blank group and a test group respectively, picking single or paired suspected escherichia coli colonies by using a sterile rod to inoculate the single or paired suspected escherichia coli colonies to one end of the test group of the identification tube, fully and uniformly mixing the colonies with a detection liquid in the biochemical identification tube, and sealing the broken opening by using sealing glue; placing the sealed identification tube in an incubator at 37 ℃, culturing for 18h, observing color changes at two ends of the biochemical tube, searching enterobacteriaceae bacteria biochemical code books (including test items such as glucose, lysine, ornithine, hydrogen sulfide, indigo substrate, lactose, dulcitol, phenylalanine, urea, citrate and the like) according to the code values, and judging the species characteristics of the strains;
the biochemical identification result shows that: the 17 strains of bacteria all accord with the basic biochemical characteristics of the escherichia coli, wherein the reaction of glucose is acid production and gas production, the reaction results of glucose, ornithine, hydrogen sulfide, indigo substrate, lactose and the like are all positive, the reaction results of lysine, dulcitol, phenylalanine, urea, citrate and the like are mostly negative, and the 17 strains of bacteria are judged to be the escherichia coli;
5) and (3) strain preservation: picking single or paired escherichia coli colonies by using a sterile rod, inoculating the escherichia coli colonies in a nutrient broth culture medium, and placing the escherichia coli colonies on a shaker for culturing at 37 ℃ and 150rpm for 18 h; adding sterilized glycerol (volume fraction is 20%) into fresh bacterial liquid according to the volume ratio of 1:1, uniformly mixing to obtain 10% of glycerol bacteria, and storing in a refrigerator at-20 ℃ for later use.
Experiments prove that the separation and identification method is accurate and reliable, the time consumption is short, and the swine escherichia coli obtained by separation is not easy to pollute.
In other embodiments of the present invention, the composition of the nutrient broth culture medium, the macanckey agar culture medium and the eosin methylene blue agar culture medium can be adjusted at will within the value range of each component, and the temperature and time of the culture can be adjusted at will within the corresponding ranges.
Claims (4)
1. A separation and identification method of Escherichia coli from swine is characterized in that: the method comprises the following steps:
1) and (3) bacteria isolation culture: inoculating the collected sample into a nutrient broth culture medium, taking a bacterium solution after culture, inoculating the bacterium solution on a Macconkey agar culture medium in a streaking manner, observing the growth condition of bacterial colonies, selecting single or paired red bacterial colonies to inoculate on an eosin methylene blue agar culture medium, observing the growth condition of the bacterial colonies, and selecting single or paired purple black bacterial colonies for later use;
2) gram staining and microscopic examination;
3) biochemical identification test;
the nutrient broth culture medium in step 1) consists of: 7-8 g of peptone, 2-2.5 g of beef extract, 3.5-4 g of sodium chloride, 750mL of distilled water and 7.0-7.4 of pH value; the composition of the Mackanka agar medium is as follows: 12-18 g of peptone, 3.5-4 g of bile salt, 3.5-4 g of sodium chloride, 10-15 g of agar, 7-8 g of lactose, 0.01-0.03 g of neutral red, 750mL of distilled water and 7.0-7.4 of pH value; the composition of eosin methylene blue agar medium is: 7.5-8.5 g of peptone, 7-8 g of lactose, 1.2-1.8 g of dipotassium hydrogen phosphate, 15-20 g of agar, 12-18 mL of 2% eosin aqueous solution, 10-15 mL of 0.5% methylene blue aqueous solution, 750mL of distilled water and 7.2-7.4 of pH value;
the gram staining operation in step 2) is as follows: fixing the selected colony smear, primarily dyeing the colony smear by using ammonium oxalate crystal violet, then mordanting the colony smear by using iodine solution, and counterdyeing the colony smear by using a re-reddening dye solution after decoloring; the microscopic examination in the step 2) is as follows: the coating area was covered with cedar oil and the staining and bacterial morphology were observed under a microscope.
2. The method of claim 1, wherein: the nutrient broth culture medium in step 1) consists of: 7.5g of peptone, 2.3g of beef extract, 3.8g of sodium chloride, 750mL of distilled water and 7.2 of pH value; the composition of the Mackanka agar medium is as follows: 15g of peptone, 3.8g of bile salt, 3.8g of sodium chloride, 12g of agar, 7.5g of lactose, 0.02g of neutral red, 750mL of distilled water and 7.2 of pH value; the composition of eosin methylene blue agar medium is: 8g of peptone, 7.5g of lactose, 1.5g of dipotassium phosphate, 18g of agar, 15mL of 2% eosin aqueous solution, 12mL of 0.5% methylene blue aqueous solution, 750mL of distilled water and 7.3 of pH value.
3. The method for separation and identification according to claim 1 or 2, wherein: the temperature for culturing in the step 1) is 36-38 ℃, and the culturing time is 15-20 h.
4. The method according to claim 1, wherein the strain biochemically identified as Escherichia coli in step 3) is inoculated into a nutrient broth, and the cultured strain is mixed with sterilized glycerol to obtain glycerol strain, and the glycerol strain is stored at a temperature below-20 ℃.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103436619A (en) * | 2013-08-27 | 2013-12-11 | 塔里木大学 | Method for identifying atypical Escherichia coli in calf diarrhea |
| CN104593476A (en) * | 2013-10-31 | 2015-05-06 | 大连大公环境检测有限公司 | An offshore escherichia coli detection method |
| CN105132519A (en) * | 2015-09-19 | 2015-12-09 | 华中科技大学 | Selective medium used for quantitative detection of escherichia coli and escherichia coli quantitative detection method |
| CN105259323A (en) * | 2015-07-22 | 2016-01-20 | 谱尼测试科技(天津)有限公司 | Novel method for determination of escherichia coli residual in chlorine-containing disinfection waste water |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103436619A (en) * | 2013-08-27 | 2013-12-11 | 塔里木大学 | Method for identifying atypical Escherichia coli in calf diarrhea |
| CN104593476A (en) * | 2013-10-31 | 2015-05-06 | 大连大公环境检测有限公司 | An offshore escherichia coli detection method |
| CN105259323A (en) * | 2015-07-22 | 2016-01-20 | 谱尼测试科技(天津)有限公司 | Novel method for determination of escherichia coli residual in chlorine-containing disinfection waste water |
| CN105132519A (en) * | 2015-09-19 | 2015-12-09 | 华中科技大学 | Selective medium used for quantitative detection of escherichia coli and escherichia coli quantitative detection method |
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