CN113999888A - Method for eluting food-borne pathogenic bacteria - Google Patents
Method for eluting food-borne pathogenic bacteria Download PDFInfo
- Publication number
- CN113999888A CN113999888A CN202111544235.3A CN202111544235A CN113999888A CN 113999888 A CN113999888 A CN 113999888A CN 202111544235 A CN202111544235 A CN 202111544235A CN 113999888 A CN113999888 A CN 113999888A
- Authority
- CN
- China
- Prior art keywords
- eluting
- food
- pathogenic bacteria
- borne pathogenic
- eluent
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- 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
- C12Q1/24—Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Pathology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for eluting food-borne pathogenic bacteria, which comprises the following steps: 1) preparing an eluent; 2) mixing an eluent and a sample in proportion, placing the mixture into an aseptic container for incubation, collecting an elution solution, wherein the eluent adopts Tris & HCl buffer solution as a basic buffer solution, and adding peptone, NaCl, cellulase and a surfactant Tween 20 to increase beating of a beating type homogenizer to improve the elution efficiency.
Description
Technical Field
The invention relates to the field of food safety, in particular to a high-efficiency elution acquisition method of food-borne pathogenic bacteria.
Background
With the development of society, the concept of balanced nutrition is gradually deepened, vegetables with low fat, low calorie and high dietary fiber are widely concerned, wherein the vegetables can be eaten fresh to reserve the nutrient components to the maximum extent, and the dishes are easy to prepare and are more and more favored. Vegetables such as lettuce, bitter chrysanthemum, cabbage, cucumber, hot pepper, onion, carrot, caraway, basil, shallot and the like can be directly eaten without heating in both traditional diet and modern diet, and more vegetables can be eaten as the vegetables are eaten together with the combination of east and west culture. However, for fresh vegetables, food-borne pathogenic bacteria can take the advantage of the fresh vegetables without heating treatment, and the fresh vegetables are harmful to the health of people.
The traditional food-borne pathogenic bacteria detection method is based on a culture method, needs a series of operations such as enrichment, selective culture medium culture, biochemical identification and the like, usually needs 4-7 days, has the defects of long time consumption and only can detect the survival in a sample, and has great limitation on fresh vegetables with high circulation and high turnover; in addition, research on a rapid detection method is always a focus of attention, but most of the existing rapid detection methods pay attention to rear-end detection, less attention is paid to links such as front-end pathogenic bacterium elution, false negative results are easy to occur if the front end cannot obtain detection target bacteria from a complex sample, and the subsequent detection steps are not helpful no matter how sensitive, so that the safety of consumers can be harmed.
Therefore, the method for detecting the pathogenic bacteria of the instant vegetables needs to be improved and improved urgently, the rapid detection without the bacteria increasing process is realized, and the false negative rate of the detection is effectively reduced.
Disclosure of Invention
In view of the above defects in the prior art, the technical problem to be solved by the invention is that the front end cannot obtain the detection target bacteria from a complex sample, and the traditional detection has the defect of long time consumption in the bacteria increasing process.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for eluting food-borne pathogenic bacteria comprises the following steps:
and 2, mixing the eluent and the sample in proportion, putting the mixture into a sterile container for incubation, and collecting the eluent as the liquid to be detected.
The step 1 comprises the following steps:
step 1.1, preparing 1-100mM Tris-HCl buffer solution as a basic buffer solution;
step 1.2, adding peptone, NaCl and a surfactant Tween 20 into the basic buffer solution in the step 1.1, and sterilizing for 15min at 121 ℃;
step 1.3, adding cellulase solution filtered by a sterile filter membrane of 0.22 mu m before use;
preferably, the pH value of the Tris-HCl buffer solution is 9-11.
Preferably, the addition amount of the peptone is 0.005-1%, the addition amount of the NaCl is 0-3%, and the addition amount of the surfactant Tween-20 is 0-0.5%.
Preferably, the final concentration of the cellulase solution after the addition of the eluent is 100-500U/L.
Preferably, the eluent prepared in step 1.2 can be prepared into a mother solution of 10X-50X, sterilized at 121 ℃ for 15min, stored and diluted into a working solution before use.
Preferably, the mass ratio of the eluent in the step 2 to the sample is 9: the mixture is put into a container with a filter screen and mixed according to the proportion of 1.
Preferably, the mass of the sample is 10-25 g.
Preferably, the incubation time is 10-60 min at room temperature.
Further, preferably, the step 2 further comprises shaking the incubated sterile homogenizing bag with the filter screen for 30min, putting the sterile homogenizing bag into a beating type homogenizer to beat and homogenize for 2min, and taking the solution on the other side of the filter screen as the solution to be detected after beating.
The invention has the advantages that:
1. the polluted food-borne pathogenic bacteria can be efficiently eluted from the vegetables through simple treatment;
2. the rapidity of detecting the food-borne pathogenic bacteria in the food sample can be ensured without increasing bacteria;
3. has universality and can carry out front-end elution aiming at various food-borne pathogenic bacteria.
Drawings
FIG. 1 is a graph comparing the elution effect of the present invention on Salmonella;
FIG. 2 is a graph showing the comparison of the elution effect of Listeria monocytogenes according to the present invention;
FIG. 3 is a graph showing the comparison of the elution effect of the present invention against Staphylococcus aureus.
Detailed Description
The technical contents of the preferred embodiments of the present invention will be more clearly and easily understood by referring to the drawings attached to the specification.
The present invention may be embodied in many different forms of embodiments and the scope of the invention is not limited to the embodiments set forth herein.
Example 1: eluting and collecting salmonella polluted in lettuce
Artificially contaminated 25g of lettuce (ATCC13076, which has rifampicin resistance after artificial induction treatment) with a contamination amount of 105CFU/g, air-dried in a clean bench and stored in a refrigerator at 4 ℃ for 1 day as a lettuce sample contaminated with Salmonella.
10mM Tris-HCl (pH9.5) buffer containing 0.9% NaCl, 0.01% peptone, 300U/L cellulase, etc. was prepared.
Adding lettuce into sterile homogenizing bag with filter screen, adding 225mL eluate, shaking for 30min, and pouring out eluate from the other side of the filter screen. The eluate was plated on LB solid medium supplemented with 100. mu.g/mL of rifampicin at 37 ℃ for counting.
Meanwhile, the same treated sample (except that the homogenizing bag is a non-filter screen homogenizing bag, other conditions are the same) is taken, the homogenate is smashed at high speed by a homogenizer, and the same volume of solution is taken for plate coating and counting to serve as a positive control.
Taking lettuce samples of the same batch, directly coating and counting on a rifampicin resistant plate after homogenization, if bacterial colonies grow, the experiment is invalid, otherwise, the experiment data is valid.
As shown in fig. 1, 1: eluting with 10mM PBS buffer (pH7.2); 2: eluting with 10mM citric acid buffer solution (pH 4.5); 3: elution with 10mM Tris-HCl buffer (pH 9.5); 4: elution with 10mM Tris-HCl buffer (pH9.5) containing 0.01% peptone; 5: elution with 10mM Tris-HCl buffer (pH9.5) containing 0.9% NaCl; 6: 10mM Tris-HCl buffer (pH9.5) containing 300U/L cellulase.
The results show that the lettuce elution rates of Tris-HCl buffer solution (pH9.5), PBS buffer solution (pH7.2) and citric acid buffer solution (pH4.5) for eluting the artificially contaminated salmonella are 41%, 53% and 64%, and obviously, the alkaline Tris-HCl buffer solution has better elution effect.
Therefore, Tris-HCl buffer (pH9.5) was used as the base buffer. On the basis, 0.01% of peptone, 0.9% of NaCl and 300U/L of cellulase are respectively added, the elution effects are 82%, 78% and 84%, and obviously, the addition of the peptone, the NaCl and the cellulase can improve the elution efficiency of the salmonella.
Example 2: highly effective elution of Listeria monocytogenes contaminated in caraway
Artificially-contaminated Lemongrass chinensis (L.) Merr with 10g of Listeria monocytogenes EGDev (with rifampicin resistance after artificial induction treatment) and with a contamination amount of about 10g5CFU/g. Air-dried in a clean bench and stored in a refrigerator at 4 ℃ for 1 day as a Salmonella-contaminated caraway sample.
Diluting 100mL of mother liquor containing 0.1% peptone in 100M Tris-HCl (pH9.5) buffer solution ten times, adjusting pH to 9.5, making up volume to 1000mL, sterilizing at 121 deg.C for 15min after preparation. After cooling, cellulase filtered by a sterile filter membrane of 0.22 mu m is added, and the final concentration of the cellulase is 300U/L. And refrigerating and storing in a refrigerator, and taking the eluate as eluent. Cellulase is added at the time of use and is used the day after the addition.
10g of caraway was added to a sterile homogenizer bag with a sieve and 90mL of eluent was added. Shaking for 30min, and homogenizing for 2min with beater homogenizer. The eluate was decanted from the other side of the filter and counted by plating at 37 ℃ onto BHI solid medium supplemented with 100. mu.g/mL rifampicin.
Meanwhile, samples treated in the same way (except that the homogenizing bag is a non-filter screen homogenizing bag, other conditions are the same) are taken, the homogenizing is smashed at high speed by a homogenizer, and the same volume of solution is taken and also subjected to plate coating counting to serve as a positive control.
Taking the same batch of caraway samples, directly coating and counting on a rifampicin resistant plate after homogenization, if bacterial colonies grow, the experiment is invalid, otherwise, the experiment data is valid.
In addition, the invention does have good elution effect for comparison, and is compared with other elution methods.
As shown in fig. 2 at 1: elution with 10mM Tris-HCl buffer (pH 9.5); 2; eluting with 10mM PBS buffer (pH7.2); 3: eluting with 10mM citric acid buffer solution (pH 4.5); 4: eluting with the mixed buffer solution; 5: beating and homogenizing commercial 0.01% peptone; 6, mixing and eluting the buffer solution, and then beating and homogenizing.
As a result, the elution of Tris-HCl buffer (pH9.5), PBS buffer (pH7.2) and citrate buffer (pH4.5) was better than that of alkaline pH9.5, but the elution of ordinary PBS buffer was less effective (only 27%).
The elution of pathogenic bacteria by directly beating and homogenizing peptone water is also a common elution mode, the elution rate is about 88 percent through comparison, and is lower than the effect of directly eluting with mixed liquor in the invention (the elution rate is 93 percent, and p is less than 0.05), namely under the condition of no beating homogenizer, the elution effect exceeding the beating and homogenizing effect of ordinary peptone water added by directly eluting with the eluent in the invention can be achieved.
After the mixed eluent is eluted, the recovery rate of about 100 percent can be achieved by homogenizing and beating, namely the most efficient elution effect is achieved.
Example 3: highly effective elution is carried out on staphylococcus aureus polluted in lettuce
10g lettuce was artificially contaminated with Staphylococcus aureus (ATCC29213, rifampicin-resistant by artificial induction) in an amount of about 105 CFU/g. Air-dried in a clean bench and stored in a refrigerator at 4 ℃ for 1 day as a lettuce sample contaminated with Staphylococcus aureus.
Diluting 100mL of mother liquor containing 0.1% peptone in 100M Tris-HCl (pH9.5) buffer solution ten times, adjusting pH to 9.5, making up volume to 1000mL, sterilizing at 121 deg.C for 15min after preparation. After cooling, cellulase filtered by a sterile filter membrane of 0.22 mu m is added, and the final concentration of the cellulase is 300U/L. And refrigerating and storing in a refrigerator, and taking the eluate as eluent. Cellulase is added at the time of use and is used the day after the addition.
10g lettuce was added to a sterile homogenizer bag with a sieve and 90mL of eluent was added. Shaking for 30min, and homogenizing for 2min with beater homogenizer. The eluate was decanted from the other side of the filter and counted by plating at 37 ℃ onto TSB solid media supplemented with 100. mu.g/mL rifampicin.
Meanwhile, samples treated in the same way (except that the homogenizing bag is a non-filter screen homogenizing bag, other conditions are the same) are taken, the homogenizing is smashed at high speed by a homogenizer, and the same volume of solution is taken and also subjected to plate coating counting to serve as a positive control.
Taking lettuce samples of the same batch, directly coating and counting on a rifampicin resistant plate after homogenization, if bacterial colonies grow, the experiment is invalid, otherwise, the experiment data is valid. In addition, the invention does have good elution effect for comparison, and is compared with other elution methods.
As shown in fig. 3, 1: eluting with pure water; 2: eluting with 10mM PBS buffer (pH7.2); 3, eluting with 10mM Tris-HCl buffer solution (pH9.5); 4: elution with 10mM Tris-HCl buffer (pH9.5) containing 0.01% glycine; 5: elution with 10mM Tris-HCl buffer (pH9.5) containing 0.01% peptone; 6: commercial 0.01% peptone water elution; 7, eluting commercialized 0.01% peptone water, beating and homogenizing; 8, mixing and eluting the buffer solution, and then beating and homogenizing.
The results showed that the lettuce-contaminated Staphylococcus aureus was eluted with the usual PBS buffer (pH7.2) at a rate of only 59%, and not even as good as the direct washing with pure water (68%). The elution rate can be improved to 81 percent by eluting with Tris-HCl with the pH value of 9.5.
In addition, the elution rate was not improved (p >0.05) by adding 0.01% glycine to Tris-HCl buffer, but was improved to 85% (p <0.05) by adding peptone (0.01%). The efficiency of beating homogenization by commercial 0.01 percent peptone water and the elution efficiency of beating homogenization after mixed incubation are 90 percent and 92 percent, while the elution efficiency of beating homogenization after mixed incubation by the mixed solution of the invention can reach 100 percent. Even in the case of no beating homogenizer, the mixed eluent of the invention is directly mixed and eluted, and the elution rate (94%) is higher than that of beating homogenization after peptone water mixed incubation.
The foregoing detailed description of the preferred embodiments of the invention has been presented. It should be understood that numerous modifications and variations could be devised by those skilled in the art in light of the present teachings without departing from the inventive concepts. Therefore, the technical solutions available to those skilled in the art through logic analysis, reasoning and limited experiments based on the prior art according to the concept of the present invention should be within the scope of protection defined by the claims.
Claims (10)
1. A method for eluting food-borne pathogenic bacteria is characterized by comprising the following steps:
step 1, preparing an eluent;
and 2, mixing the eluent and the sample in proportion, putting the mixture into a sterile container for incubation, and collecting the eluent.
2. The method for eluting food-borne pathogenic bacteria according to claim 1, wherein the step 1 comprises the following steps:
step 1.1, preparing 1-100mM Tris-HCl buffer solution as a basic buffer solution;
step 1.2, adding peptone, NaCl and a surfactant Tween 20 into the basic buffer solution in the step 1.1, and sterilizing for 15min at 121 ℃;
step 1.3, cellulase solution filtered through a 0.22 μm sterile filter before use.
3. The method for eluting food-borne pathogenic bacteria according to claim 2, wherein the pH value of the Tris-HCl buffer solution is 9-11.
4. The method for eluting food-borne pathogenic bacteria according to claim 2 or 3, wherein the addition amount of the peptone is 0.005-1%, the addition amount of the NaCl is 0-3%, and the addition amount of the surfactant Tween 20 is 0-0.5%.
5. The method for eluting food-borne pathogenic bacteria according to any one of claims 2 to 4, wherein the final concentration of the cellulase solution after the addition of the eluent is 500U/L.
6. The method for eluting food-borne pathogenic bacteria according to claim 2, wherein the eluent obtained in the step 1.2 is prepared into 10 x-50 x mother liquor, and the mother liquor is sterilized at 121 ℃ for 15min, stored and diluted into working solution before use.
7. The method for eluting food-borne pathogenic bacteria according to claim 1, wherein the elution solution obtained in the step 2 is mixed with the sample according to a mass ratio of 9: the mixture is mixed in a sterile container with filter holes according to the proportion of 1.
8. The method for eluting food-borne pathogenic bacteria according to claim 1 or 7, wherein the sample mass is 10-25 g.
9. The method for eluting food-borne pathogenic bacteria according to claim 1, wherein the incubation time is 10-60 min at room temperature.
10. The method for eluting food-borne pathogenic bacteria according to claim 1, wherein the step 2 comprises shaking the incubated sterile homogenizer bag with a filter screen for 30min, placing the homogenizer in a beating type homogenizer to beat and homogenize for 2min, and taking the solution outside the container as the solution to be tested after beating.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111544235.3A CN113999888A (en) | 2021-12-16 | 2021-12-16 | Method for eluting food-borne pathogenic bacteria |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111544235.3A CN113999888A (en) | 2021-12-16 | 2021-12-16 | Method for eluting food-borne pathogenic bacteria |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113999888A true CN113999888A (en) | 2022-02-01 |
Family
ID=79931783
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111544235.3A Pending CN113999888A (en) | 2021-12-16 | 2021-12-16 | Method for eluting food-borne pathogenic bacteria |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113999888A (en) |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101570783A (en) * | 2009-03-20 | 2009-11-04 | 郑秋月 | Detection kit and detection method for 9 species of pathogenic organisms in marine products |
| CN101748176A (en) * | 2008-12-08 | 2010-06-23 | 中华人民共和国黑龙江出入境检验检疫局检验检疫技术中心 | High-efficiency extracting method of food-borne pathogen nucleic acid |
| CN103160577A (en) * | 2013-02-05 | 2013-06-19 | 上海交通大学 | Method for rapidly detecting pathogenic bacteria in food |
| CN106521031A (en) * | 2016-11-30 | 2017-03-22 | 浙江省疾病预防控制中心 | Method for detecting norovirus in food based on negatively charged membrane concentration |
| CN106896225A (en) * | 2017-04-28 | 2017-06-27 | 河南艾能生物科技有限公司 | A kind of method for quick of salmonella |
| CN109541205A (en) * | 2018-12-07 | 2019-03-29 | 河南省商业科学研究所有限责任公司 | The detection method of lactic acid bacteria in a kind of probiotics |
| CN109666590A (en) * | 2019-01-29 | 2019-04-23 | 大连民族大学 | A kind of method of microorganism in collection fresh-cut fruit and vegetable |
| CN112080588A (en) * | 2020-09-10 | 2020-12-15 | 上海市农业科学院 | Method for detecting norovirus in fruits and vegetables |
-
2021
- 2021-12-16 CN CN202111544235.3A patent/CN113999888A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101748176A (en) * | 2008-12-08 | 2010-06-23 | 中华人民共和国黑龙江出入境检验检疫局检验检疫技术中心 | High-efficiency extracting method of food-borne pathogen nucleic acid |
| CN101570783A (en) * | 2009-03-20 | 2009-11-04 | 郑秋月 | Detection kit and detection method for 9 species of pathogenic organisms in marine products |
| CN103160577A (en) * | 2013-02-05 | 2013-06-19 | 上海交通大学 | Method for rapidly detecting pathogenic bacteria in food |
| CN106521031A (en) * | 2016-11-30 | 2017-03-22 | 浙江省疾病预防控制中心 | Method for detecting norovirus in food based on negatively charged membrane concentration |
| CN106896225A (en) * | 2017-04-28 | 2017-06-27 | 河南艾能生物科技有限公司 | A kind of method for quick of salmonella |
| CN109541205A (en) * | 2018-12-07 | 2019-03-29 | 河南省商业科学研究所有限责任公司 | The detection method of lactic acid bacteria in a kind of probiotics |
| CN109666590A (en) * | 2019-01-29 | 2019-04-23 | 大连民族大学 | A kind of method of microorganism in collection fresh-cut fruit and vegetable |
| CN112080588A (en) * | 2020-09-10 | 2020-12-15 | 上海市农业科学院 | Method for detecting norovirus in fruits and vegetables |
Non-Patent Citations (2)
| Title |
|---|
| RONALD H等主编，石阶平等译: "食品安全手册", pages: 112 * |
| 朱金艳等: "果蔬中诺如病毒检测技术研究进展", 《中国果菜》, vol. 41, no. 3, 15 March 2021 (2021-03-15), pages 43 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Whyte et al. | Quantitative investigation of the effects of chemical decontamination procedures on the microbiological status of broiler carcasses during processing | |
| Evrendilek et al. | Inactivation of Escherichia coli O157: H7 and Escherichia coli 8739 in apple juice by pulsed electric fields | |
| Farber et al. | Thermal resistance of Listeria monocytogenes in inoculated and naturally contaminated raw milk | |
| Hao et al. | Efficacy of slightly acidic electrolyzed water (SAEW) for reducing microbial contamination on fresh‐cut cilantro | |
| Samelis et al. | Influence of the natural microbial flora on the acid tolerance response of Listeria monocytogenes in a model system of fresh meat decontamination fluids | |
| Li et al. | Dual effects of acidic electrolyzed water treatments on the microbial reduction and control of enzymatic browning for fresh‐cut lotus root | |
| Oko et al. | Bacteriological and nutritional analysis of groundnut cake sold in an open market in Samaru, Zaria-Kaduna State | |
| Moosavy et al. | Isolation of Listeria monocytogenes from milks used for Iranian traditional cheese in Lighvan cheese factories | |
| CA3015072A1 (en) | Reduced negative zeta potential sanitization method and system | |
| CN105724985B (en) | Juice modification method that is a kind of while delaying ascorbate degradation and removal heavy metal lead | |
| CN113999888A (en) | Method for eluting food-borne pathogenic bacteria | |
| CN104248012A (en) | Concentrated Noni Juice and processing technology thereof | |
| Abid et al. | Microbial quality assessment study of branded and unbranded milk sold in Peshawar City, Pakistan | |
| CN105495483A (en) | Method for making bacteriostatic-agent dry powder with pickle brine | |
| Khudhir et al. | The efficacy of ozone treatment on the microbiological quality of raw milk at different storage temperatures | |
| Mamun et al. | Prevalence of foodborne pathogens and effectiveness of washing or cooking in reducing microbiological risk of contaminated red amaranth | |
| CN105029521B (en) | The processing method for eating octopus conditioning food raw | |
| Nzabuheraheza et al. | Microbiological analysis of traditionally fermented milk sold in Kinigi Sector of Musanze District in Rwanda | |
| CN107034159A (en) | A kind of pathogenic bacteria antagonist and its application | |
| Mukhopadhyay et al. | An evaluation of street-vended sliced papaya (Carica papaya) for bacteria and indicator micro-organisms of public health significance | |
| Olasupo et al. | Evaluation of Nisin for the preservation of Nono‐A Nigerian Fermented Milk Product | |
| Singh et al. | Isolation of food pathogenic bacteria from unhygienic fruit juice mill and screening various herbal plant extracts for inhibitory potential | |
| Obi et al. | Bacteriological analyses of weaning foods consumed in Umuariaga community, Ikwuano LGA Abia state | |
| JP5368170B2 (en) | Method for producing antibacterial ionized metal composition | |
| AU2021103584A4 (en) | Method for cold chain sterilization and disinfection |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |