CN1077750A - Food micro-organism detecting plate and preparation thereof and application - Google Patents
Food micro-organism detecting plate and preparation thereof and application Download PDFInfo
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- CN1077750A CN1077750A CN 93105833 CN93105833A CN1077750A CN 1077750 A CN1077750 A CN 1077750A CN 93105833 CN93105833 CN 93105833 CN 93105833 A CN93105833 A CN 93105833A CN 1077750 A CN1077750 A CN 1077750A
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Abstract
The present invention relates to a kind of food bacteria total number and coliform check-out console, this plate can once be finished the detection of total plate count and coliform simultaneously.The invention still further relates to the preparation and the application of this check-out console.
Description
The present invention relates to a kind of food bacteria total number and coliform check-out console, its preparation method and application.
Food often is subjected to microbiological contamination in the external environment in processes such as processing, transportation, preservation and sale.Food bacterial contamination can cause putrid and deteriorated, even causes food poisoning, is one of modal adverse factor in the food sanitation." food hygienic standard " regulation of China's promulgation, the routine monitoring of food sanitation requires check total plate count and coliform, though national standard method is accurate, the operation more complicated, it is long to detect the time that needs.In recent years, the fast simple detection method of domestic some that work out, can accelerate detection time in varying degrees, simplify procedures, but in these methods, what have can only the bacterial detection sum, and what have can only detect coliform, all can not finish the check of above-mentioned two indexs simultaneously, and some method is only at certain based food.
In addition, coliform means that a group energy produces sour aerogenesis, aerobic amphimicrobian Gram-negative sporeless bacterium at 36 ℃ of ferment lactosees in 24 hours, in fact comprises four kinds of common bacterias in the enterobacteriaceae: colon bacillus (E.coli), expense labor Di Shi citric acid bacillus (C.freundii), Klebsiella pneumonia (K.pneumoniae), enterobacter cloacae (E.cloacae).So coliform is not single bacterium, but bacterioid of selecting as sanitary index.More existing detection substratum only are used to detect a certain bacterium, as special detection colon bacillus (E.coli).
For satisfying the needs of food hygiene detection, we develop a kind of food simple detection plate that can detect food bacteria total number and coliform simultaneously.The employed substratum of this check-out console is wider than the substratum sensing range of prior art to the scope that coliform detects.And the employed substratum of check-out console of the present invention adopts the proportioning and the content of special component, thereby reaches in the detection the selectivity of different monoid bacteriums or comprehensive.
Technology in the past (comprising national standard method) adopts diverse ways respectively for detecting total count and coliform group count, two kinds of detection methods may not made up, and the present invention is when simplifying two kinds of detection methods respectively, for these two kinds of combinations provide possibility.
That check-out console of the present invention can make is easy and simple to handle, the result fast, expense economy and accuracy reached national standard.
The object of the present invention is to provide a kind of food simple detection plate that can detect food bacteria total number and coliform simultaneously.
Another object of the present invention is to provide the method for the above-mentioned check-out console of preparation.
A further object of the invention provides the application that detects food bacteria total number and coliform with this check-out console simultaneously.
Check-out console of the present invention is a rectangular slab, and its size can be 5.0 * 3.6 * 0.2cm, to being divided into two portions, is respectively charged into substratum 1 and substratum 2(Fig. 1 by the central longitudinal of plate), this plate one side can have a hand-held handle (Fig. 2).Should be appreciated that as one of skill in the art the variation of this check-out console size shape is a general knowledge, therefore any modifications and changes for its size and shape include within the scope of the present invention.
Substratum 1 is to be used for the substratum that total plate count detects, and substratum 2 is used to measure coliform, and it forms with the preparation method as follows.
Substratum 1
Component content
The immersion liquid of ox brain
*150～250 milliliters
The OX-heart immersion liquid
*150～250 milliliters
Peptone 10 grams
Glucose 2 grams
Sodium-chlor 5 grams
Dipotassium hydrogen phosphate 2.5 grams
Agar 15 grams
Distilled water adds to 1000 milliliters
PH7.0-7.4
* brain-heart-infusion: OX-heart and each 500 gram of ox brain with fresh degrease and manadesma rub, place 2000 milliliters of Erlenmeyer flasks respectively, 1000 milliliters of mixings of each adding distil water are placed on refrigerator overnight.Next day, skim oil slick with casserole respectively, be placed in 45 ℃ of water-baths 1 hour again, heated and boiled is 30 minutes then, and filtered through gauze is mended respectively to 1000 milliliters with distilled water then, 15 pounds of sterilizations in 20 minutes.
Will be by 15 pounds of heat sterilizations of substratum of above-mentioned formulation 15 minutes, about sterilization postcooling to 50 ℃.Add 5mlTTC(RT, 5% aqueous solution by every 100ml substratum) ratio add TTC, mixing.
In substratum 1, preferably contain brain immersion liquid of 200ml ox and the immersion liquid of 250ml OX-heart.
Substratum 2
Component content
Extractum carnis
*3 grams
Peptone 20 grams
Lactose 10 grams
Sodium deoxycholate 1.0 grams
Sulfothiorine 2.3 grams
Trisodium Citrate 1.0 grams
Ferric ammonium citrate 1.0 grams
Toluylene red 0.03 gram
Agar 15 grams
Distilled water adds to 1000 milliliters
PH6.8～7.0
* extractum carnis: remove its fat, sarolemma and tendon with fresh beef, add twice (weight) water logging after the rubbing and steep a liquid, boil after-filtration, solidifying egg white is removed, become meat extract, make through 70 ℃ of vacuum-evaporation.
With 15 pounds by the substratum heat sterilization of above-mentioned formulation 15 minutes.
The experiment and the result that detect food bacteria total number and coliform with check-out console of the present invention are as follows.
Material
1. bacterial strain
Pseudomonas aeruginosa (Pseudomonas aeruginosa), flavobacterium odoratum (Flavobacterium odoratum), kafir lily suis (Streptococcusmimiatus), xanthobacter autotrophicus (Xanthobacter autotrophicus), plant living klebsiella (Klebsiellaplanticola), Bacterium lacticum (Lactobacillus sp.), have a liking for Fructus Hordei Germinatus pseudomonas (Pseudomoenasmaltophilia), Bacillus licheniformis (Bacillus licheniformis), colon bacillus (Escherichia coli), subtilis (Bacillus subtilis), bacillus pumilus (Bacillus brevis), (above provide) and Klebsiella pneumonia (Klebsiella pneumoniae) (providing) by Heilongjiang Prov. Sanitation and Antiepidemic Station by Institute of Microorganism, Academia Sinica.
2. substratum
(1) total plate count substratum (above-mentioned substratum 1);
(2) coliform substratum (above-mentioned substratum 2);
(3) gravy peptone;
(4) PYG substratum.
Method
1. substratum selectivity, specificity and stability experiment
Substratum is made flat board, after the surface drying,, cultivate after 24 hours and 48 hours for 30 ℃ and observe, do contrast with gravy peptone and PYG substratum simultaneously with liquid culture one platinum loop inoculation in 24 hours.
With preparation put 4 ℃ of freezer storages through 10 qualified check-out consoles of sterility test, take out after 1 month, 3 months and half a year, after the inoculation of colon bacillus suspension diluent, cultivated 24 hours, and, calculated the coincidence rate of two methods for 37 ℃ with National Standard Method controlled observation result.
2. preliminary experiment
(1) gets 24 hours nutrient agar medium slant cultures of colon bacillus, contain the bacteria suspension of 800,400,200,100,50,20 bacteriums in being made into every milliliter with sterile saline.
(2) check-out console is dipped in the bacteria suspension of above-mentioned each concentration, blotted unnecessary bacterium liquid with sterilization filter paper then, cultivated the colony number that grows on the gauging board 18～24 hours for 37 ℃.Simultaneously, each standard bacteria suspension as a sample to be checked, is used the MPN value of jus gentium bacterial detection sum and coliform respectively.
(3) relation conefficient of calculating check-out console method and National Standard Method is analyzed correlationship.
(referring to " vital statistics ", the 2nd edition, willow duty, People's Health Publisher, 1985, the 103-109 pages or leaves.）
3. formal experiment
By National Standard Method sample is done pre-treatment, the MPN value of bacterial detection sum and coliform.Simultaneously, each check-out console is dipped in the dilution sample of difference, blotted unnecessary liquid, cultivated 18～24 hours, and write down its colony number for 37 ℃ with sterilization filter paper.
4. the statistical procedures of experimental result
The total plate count that National Standard Method is recorded and the MPN value of coliform.With the colony number input computer that grows on the check-out console of corresponding 10 times of dilutions, leave out insincere data by 95% fiducial limit, do the correlationship analysis respectively, draw linear regression equation (referring to " vital statistics ", as mentioned above).
5. the preparation of on-gauge plate
According to the linear regression equation that draws, and calculate colony number on the different pollution level on-gauge plates of following representative.
Below the not contaminated national normal value
Edible pollution national normal value is below 5 times
In 25 times of the slight pollution national normal values
In 125 times of the intermediate pollution national normal values
The serious pollution national normal value is more than 125 times
6. examination is used
In Shijiazhuang, the health and epidemic prevention station on ground such as Huhehaote, Shenyang, Harbin, Qiqihar, Hailaer, 1058 parts of samples are detected with National Standard Method and check-out console method simultaneously.
Discussion of results
1. the growing state of various bacteriums on two kinds of substratum.
With four kinds of substratum of known inoculation, 30 ℃ of growing states after 24～48 hours see Table 1.
The comparison of the various bacteriums of table 1 growing state on substratum
Annotate: +～growth; Not-～do not grow; +-～poor growth.
The bacterium that can not grow on gravy peptone and PYG substratum can grow on the check-out console method is used for the substratum of bacterial detection sum.Simultaneously, dysgonic flora can fine growth.The desaturase of bacterium makes the TTC reduction form red bacterium colony, and quality is faint yellow, is easy to identification and counting.
Be used to detect the check-out console substratum of coliform, owing to contain inhibitor and indicator, the bacterium of the coliform of only growing, and the pink colour and the reddish black bacterium colony of formation feature, the quality sorrel, mycetozoan is not filled the air growth.
2. the result of preliminary experiment shows, the relation conefficient of check-out console method and National Standard Method bacterial detection sum and coliform is respectively 0.96 and 0.98, and degree of correlation is very high, thereby provides scientific basis for experimental applications.
3. with 340 parts of check-out console test sample, learn by statistics and handle, the data in its 95% fiducial limit, total plate count is 314 parts, coliform is 322 parts.The gained data are done scatter diagram, and it is linear that the result is, and linearly correlationship of check-out console method and National Standard Method is described.Its relation conefficient and linear regression equation such as table 2.
The correlation regression relation of table 2 check-out console method and National Standard Method bacterial detection sum and coliform value
Test item total plate count coliform value
Relation conefficient 0.9926 0.9999
Linear regression equation Y=-5008+208X Y=-51+45X
* the Y-National Standard Method records and total plate count and MPN value.
Total plate count that X-check-out console method records and coliform number.
The The above results explanation, the degree of correlation of two methods is very high, and the credible result of check-out console method can replace National Standard Method to estimate the pollution condition of food.In addition, from above-mentioned two kinds of regression equations, find out, the substratum of check-out console is relatively more responsive, with regard to measuring total plate count, the result＜1(that records with National Standard Method is 0=-5008+208X, X=24～25), 24～25 bacterium colonies of but growing on the platelet of our development, the bacterium that some is not grown on the ordinary nutrient agar substratum, on the substratum that the check-out console method is used, also can grow and the growth experiment result consistent.
The detected result of coliform also is like this.If the MPN value that records with National Standard Method is 30, be coliform group count＜1 in every milliliter of sample, and the inoculum size of check-out console only is about 0.2ml, so infers the bacterial growth that do not have on the platelet, but in fact grow 1～2 bacterium colony, this is because the nutritional condition of substratum cause preferably.Though the substratum sensitivity of check-out console can detect the bacterium that National Standard Method can not detect, we are to be the regression equation that draws of contrast with National Standard Method, and it combines the area of check-out console, and all multifactor influences such as inoculum size are so detected result is believable.
4. according to the food hygienic standard of China and the regression equation of experiment gained, the expression scope such as the table 3 of the plate that settles the standard.
The expression scope of table 3 food bacteria total number and coliform examination criteria tally
Although the hygienic standard difference of various food, the on-gauge plate that we make is unified, because balance in addition from the dilution number of sample; Total plate count and the low sample of coliform MPN value in the standard, extension rate is relatively little, otherwise will increase relatively.The concrete extension rate of various food is seen attached list.In testing, as long as sample is done suitable dilution, the direct inoculation check-out console, after cultivating in 18～24 hours, the bacterium colony and the on-gauge plate of growth are contrasted, can infer the approximate range of total plate count and coliform MPN value, the pollution level of the tested food of intuitive judgment.
5. with the National Standard Method contrast, food bacteria total number and coliform check-out console with our development detect 1058 parts of varieties of food items altogether, and two method gained results' coincidence rate reaches more than 97% (table 4).10 check-out consoles making took out from 4 ℃ of refrigerators after 1 month, 3 months and half a year, inoculated the colon bacillus diluent, except that a platelet because of the contaminant, the result that other platelets record all conforms to, and illustrates that the substratum quality of check-out console use is stable, and susceptibility does not have change.
The result that above presentation of results check-out console method is measured need not prepare substratum in addition and scrub sterilization equipment accurately and reliably, and has simple to operately, easy to use, time saving and energy saving, is easy to advantages such as judgement, is applicable to vast grass-roots unit.
The recombination rate of table 41058 part food sample two method detected results
Claims (6)
1, can detect the food simple detection plate of food bacteria total number and coliform simultaneously, it is characterized in that this plate is a rectangular slab, to being divided into two portions, being respectively charged into the substratum of bacterial detection sum and detecting the substratum of coliform by the central longitudinal of plate.
2, check-out console according to claim 1, the substratum of wherein said bacterial detection sum is made up of following ingredients:
Component content
150～250 milliliters of ox brain immersion liquid
150～250 milliliters of OX-heart immersion liquid
Peptone 10 grams
Glucose 2 grams
Sodium-chlor 5 grams
Dipotassium hydrogen phosphate 2.5 grams
Agar 15 grams
Distilled water adds to 1000 milliliters
PH7.0-7.4
5% aqueous solution that in per 100 milliliters of above-mentioned substratum, also contains 5 milliliters of RTs.
3, check-out console according to claim 1, the substratum of wherein said detection coliform is made up of following ingredients:
Component content
Extractum carnis 3 grams
Peptone 20 grams
Lactose 10 grams
Deoxidation depickling sodium 1.0 grams
Hypo 2.3 grams
Trisodium Citrate 1.0 grams
Ferric ammonium citrate 1.0 grams
Toluylene red 0.03 gram
Agar 15 grams
Distilled water adds to 1000 milliliters
PH6.8-7.0
4, check-out console according to claim 1, wherein check-out console is the rectangular slab of 5.0 * 3.6 * 0.2cm size.
5, check-out console according to claim 1 also has a hand-held handle in the one side.
6, detect the application of food bacteria total number and coliform simultaneously with the described check-out console of claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 93105833 CN1077750A (en) | 1993-05-25 | 1993-05-25 | Food micro-organism detecting plate and preparation thereof and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 93105833 CN1077750A (en) | 1993-05-25 | 1993-05-25 | Food micro-organism detecting plate and preparation thereof and application |
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| Publication Number | Publication Date |
|---|---|
| CN1077750A true CN1077750A (en) | 1993-10-27 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1064707C (en) * | 1997-01-31 | 2001-04-18 | 黑龙江省轻工业研究所 | Method for prodn. of convenient microbe substratum |
| CN102212604A (en) * | 2010-04-12 | 2011-10-12 | 无锡市华商生物试剂有限公司 | Intestinal enriched tablet culture medium and process method thereof |
| WO2019015357A1 (en) * | 2017-07-21 | 2019-01-24 | 深圳市伯劳特生物制品有限公司 | Medium |
| CN109576338A (en) * | 2018-11-21 | 2019-04-05 | 广东顺德清宇环保科技有限公司 | A kind of microorganism color developing agent and preparation method thereof and viable count detection device and preparation method thereof and detection method |
-
1993
- 1993-05-25 CN CN 93105833 patent/CN1077750A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1064707C (en) * | 1997-01-31 | 2001-04-18 | 黑龙江省轻工业研究所 | Method for prodn. of convenient microbe substratum |
| CN102212604A (en) * | 2010-04-12 | 2011-10-12 | 无锡市华商生物试剂有限公司 | Intestinal enriched tablet culture medium and process method thereof |
| CN102212604B (en) * | 2010-04-12 | 2014-07-23 | 无锡市华商生物试剂有限公司 | Intestinal enriched tablet culture medium and process method thereof |
| WO2019015357A1 (en) * | 2017-07-21 | 2019-01-24 | 深圳市伯劳特生物制品有限公司 | Medium |
| CN109576338A (en) * | 2018-11-21 | 2019-04-05 | 广东顺德清宇环保科技有限公司 | A kind of microorganism color developing agent and preparation method thereof and viable count detection device and preparation method thereof and detection method |
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