GB 4789.7-2013_English: PDF (GB4789.7-2013)
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National Food Safety Standard -- Food Microbiological Examination -- Vibrio parahaemolyticus
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Microbiological examination of food hygiene -- Examination of vibrio parahaemolyticus
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Microbiological examination of food hygiene -- Examination of Vibrio parahaemolyticus
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Microbiological examination of food hygiene. Examination of Vibrio parahaemolyticus
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Microbiological examination of food hygiene--Examination of vibrioparahaemolyticus
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| Standard ID | GB 4789.7-2013 (GB4789.7-2013) | | Description (Translated English) | National Food Safety Standard-Food Microbiological Examination - Vibrio parahaemolyticus | | Sector / Industry | National Standard | | Classification of Chinese Standard | C53 | | Classification of International Standard | 07.100.30 | | Word Count Estimation | 17,161 | | Older Standard (superseded by this standard) | GB 4789.7-2008 | | Drafting Organization | National Food Industry Standardization Technical Committee | | Administrative Organization | National Health & Family Planning Commission of PRC | | Regulation (derived from) | China Food & Drug Administration [2013] No. 234, November, 1, 2013 | | Issuing agency(ies) | National Health and Family Planning Commission of the People's Republic of China | | Summary | This Standard, from June 1, 2014, substitutes GB/T 4789. 7-2008 Food Microbiological Examination Vibrio parahaemolyticus. | | Standard ID | GB/T 4789.7-2008 (GB/T4789.7-2008) | | Description (Translated English) | Microbiological examination of food hygiene. Examination of vibrio parahaemolyticus | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | C53 | | Classification of International Standard | 07.100.30 | | Word Count Estimation | 14,168 | | Date of Issue | 2008-05-16 | | Date of Implementation | 2008-11-01 | | Older Standard (superseded by this standard) | GB/T 4789.7-2003 | | Adopted Standard | FDA Chapter 9, MOD | | Drafting Organization | China Disease Prevention and Control Center of Nutrition and Food Safety | | Administrative Organization | Ministry of Health | | Regulation (derived from) | National Standard Approval Announcement 2008 No.9 (Total No.122); the National Health and Family Planning Commission, 2013 No.7 Announcement | | Proposing organization | Ministry of Health of the People Republic of China | | Issuing agency(ies) | The People's Republic of China Ministry of Health, China National Standardization Management Committee | | Summary | This standard specifies the test methods in food Vibrio parahaemolyticus. This standard applies to fish and food poisoning Vibrio parahaemolyticus sample test, can refer to the use of other foods. | | Standard ID | GB/T 4789.7-2003 (GB/T4789.7-2003) | | Description (Translated English) | Microbiological examination of food hygiene. Examination of Vibrio parahaemolyticus | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | C53 | | Classification of International Standard | 07.100.30 | | Word Count Estimation | 5,549 | | Date of Issue | 2003/8/11 | | Date of Implementation | 2004/1/1 | | Older Standard (superseded by this standard) | GB/T 4789.7-1994 | | Quoted Standard | GB/T 4789.28-2003 | | Regulation (derived from) | Announcement of Newly Approved National Standards No. 9 in 2008 (No. 122 overall) | | Proposing organization | Ministry of Health of the People Republic of China | | Issuing agency(ies) | People Republic of China Ministry of Health China National Standardization Management Committee |
GB 4789.7-2013
GB
NATIONAL STANDARD OF
THE PEOPLE’S REPUBLIC OF CHINA
National Food Safety Standard - Food Microbiological
Examination - Vibrio parahaemolyticus
ISSUED ON. NOVEMBER 29, 2013
IMPLEMENTED ON. JUNE 1, 2014
Issued by. National Health and Family Planning Commission of China
Table of Contents
Foreword ... 3
1 Scope ... 4
2 Normative references ... 4
3 Mediums and reagents ... 4
4 Inspection procedures ... 5
5 Requirements ... 6
6 Serological typing (optional) ... 8
7 Kanagawa test (optional) ... 10
8 Results and reports ... 10
Annex A Medias and reagents ... 12
Annex B Vibrio parahaemolyticus most probable number (MPN) search table19
National Food Safety Standard - Food Microbiological
Examination - Vibrio parahaemolyticus
1 Scope
This Standard specifies the inspection method for Vibrio parahaemolyticus in
food.
This Standard is applicable to the inspection of Vibrio parahaemolyticus in
food.
2 Normative references
In addition to microbial laboratory routine sterilization and culture equipment,
other equipment and materials are as follows.
a) constant temperature incubator. 36°C ± 1°C;
b) refrigerator. 2°C ~ 5°C, 7°C ~ 10°C;
c) constant temperature water bath. 36°C ± 1°C;
d) homogenizer or sterile mortar;
e) balance. resolution of 0.1 g;
f) sterile test tubes. 18mm × 180mm, 15mm × 100mm;
g) sterile straws. 1 mL (with 0.01 mL scale), 10 mL (with 0.1 mL scale) or
micro-pipettes and tips;
h) sterile conical flasks. volumes of 250 mL, 500 mL, 1000 mL;
j) sterile Petri dish. diameter of 90 mm;
j) automatic microbial biochemical identification system;
k) sterile surgical scissors, tweezers.
3 Mediums and reagents
3.1 3% sodium chloride alkaline peptone water. see A.1 of Annex A
3.2 thiosulfate-citrate-bile salt-sucrose (TCBS) agar. see A.2 of Annex A
3.3 3% sodium chloride trypsin soybean agar. see A.3 of Annex A
3.4 3% sodium chloride trisaccharide iron agar. see A.4 of Annex A
3.5 salt test medium. see A.5 of Annex A
3.6 3% sodium chloride mannitol test medium. see A.6 of Annex A
3.7 3% sodium chloride lysine decarboxylase test medium. see A.7 of
Annex A
3.8 3% sodium chloride MR-VP medium. see A.8 of Annex A
3.9 3% sodium chloride solution. see A.9 of Annex A
3.10 Wagstsuma blood agar. see A.10 of Annex A
3.11 oxidase reagents. see A.11 of Annex A
3.12 gram stain solution. see A.12 of Annex A
3.13 ONPG. see A.13 of Annex A
3.14 Voges-Proskauer (V-P) reagent. see A.14 of Annex A
3.15 vibrio color development medium
3.16 biochemical identification kit
4 Inspection procedures
See Figure 1 for Vibrio parahaemolyticus test procedures.
5.4 Pure culture
Pick up three or more suspicious colonies. Scribe 3% sodium chloride trypsin
soybean agar plate. Cultivate at 36°C ± 1°C for 18h ~ 24h.
5.5 Preliminary identification
5.5.1 Oxidase test. select a single culture of pure culture for oxidase test;
Vibrio parahaemolyticus shall be oxidase positive.
5.5.2 Smear microscopy. smear the suspected colony; carry out Gram
stain; perform the microscopic examination on observation morphology.
Vibrio parahaemolyticus shall be Gram-negative, rod-shaped, curved, oval,
without spores, with flagella.
5.5.3 Pick purely cultured single suspicious colony. Transfect 3% sodium
chloride trisaccharide agar slope and puncture the bottom layer. Cultivate at
36°C ± 1°C for 24h to observe the results. The reaction of Vibrio
parahaemolyticus in 3% sodium chloride trisaccharide iron agar shall be that
the bottom turns yellow not black, without bubbles; slope color shall be
unchanged or red becomes deeper, with power.
5.5.4 Halophilic test. pick purely cultured single suspicious colony;
respectively inoculate 0%, 6%, 8% and 10% peptone water of different
concentrations of sodium chloride; cultivate at 36°C ± 1°C for 24h observation;
observe the liquid turbidity. Vibrio parahaemolyticus shall not grow or grow
weakly in cisplatin without sodium chloride and of 10% sodium chloride. It
shall grow vigorously in peptone water of 6% sodium chloride and 8% sodium
chloride.
5.6 Determination of identification
Take pure culture and respectively inoculate mannitol test medium containing
3% sodium chloride, lysine decarboxylase test medium, MR-VP medium.
Observe the results after cultivating at 36°C ± 1°C for 24h ~ 48h. Use 3%
sodium chloride trisaccharide agar overnight culture for ONPG test. It can
select biochemical identification kit or automatic microbial biochemical
identification system.
6 Serological typing (optional)
6.1 Preparation
Inoculate two tubes of 3% sodium chloride trypsin soy agar test tube slope.
Cultivate at 36°C ± 1°C for 18h ~ 24h. Use 5% glycerol solution containing 3%
sodium chloride to wash 3% sodium chloride trypsin soy agar slant culture so
Distilled water 1000.0 mL
A.3.2 Method
Dissolve the ingredients in A.3.1 in distilled water. Correct pH to 7.3 ± 0.2.
Sterilize at high temperature of 121°C for 15 min.
A.4 3% sodium chloride trisaccharide iron agar
A.4.1 Ingredient
Peptone 15.0 g
Proteose peptone 5.0 g
Beef jelly 3.0 g
Yeast extract 3.0 g
Sodium chloride 30.0 g
Lactose 10.0 g
Sucrose 10.0 g
Glucose 1.0 g
Ferrous sulfate (FeSO4) 0.2 g
Phenol red 0.024 g
Sodium thiosulfate (Na2S2O3) 0.3 g
Agar 12.0 g
Distilled water 1000.0 mL
A.4.2 Method
Dissolve the ingredients in A.4.1 in distilled water. Correct pH to 7.4 ± 0.2.
Sub-fill into the test tubes of appropriate capacities. Sterilize at high
temperature of 121°C for 15 min. Made high-level slope of which the slope
length is 4 cm ~ 5 cm; high depth is 2 cm ~ 3 cm.
A.5 Salt test medium
A.5.1 Ingredient
Tryptone 10.0 g
Sodium chloride Add in different amounts
Distilled water 1000.0 mL
A.5.2 Method
Dissolve the ingredients in A.5.1 in distilled water. Correct pH to 7.2 ± 0.2.
Prepare 5 bottles in total, 100 mL of each bottle. Each bottle shall be added
with varying amounts of sodium chloride. (1) none; (2) 3 g; (3) 6 g; (4) 8 g; (5)
10 g. Sub-fill into the test tubes. Sterilize at high temperature of 121°C for 15
min.
A.6 3% sodium chloride mannitol test medium
Dissolve N, N, N ', N'-tetramethyl-p-phenylenediamine hydrochloride in
distilled water. Store in 2°C ~ 5°C refrigerator from light. Use within 7d.
A.11.3 Test method
Use fine glass rods or one-time inoculation needle to select fresh (24 h)
colony. Coat on a filter paper moistened by oxidase reagents. If the filter
paper within 10 s shows pink or purple, it shall be positive for oxidase test. No
discoloration shall be negative for oxidase test.
A.12 Gram stain solution
A.12.1 Crystal violet dyeing solution
A.12.1.1 Ingredient
Crystal violet 1.0 g
95% ethanol 20.0 mL
1% aqueous ammonium oxalate solution 80.0 mL
A.12.1.2 Method
Completely dissolve the crystal violet in ethanol and then mix with ammonium
oxalate solution.
A.12.2 Gram iodine solution
A.12.2.1 Ingredient
Iodine 1.0 g
Potassium iodide 2.0 g
Distilled water 1000.0 mL
A.12.2.2 Method
Mix iodine with potassium iodide. Add distilled water to shake a little. After
being completely dissolved, add distilled water to 300 mL.
A.12.3 Safranin complex liquid
A.12.3.1 Ingredient
Safranin 0.25 g
95% ethanol 10.0 mL
Distilled water 1000.0 mL
A.12.3.2 Method
Dissolve safranin n ethanol and then dilute with distilled water.
A.12.4 Dyeing method
......
GB/T 4789.7-2008
Microbiological examination of food hygiene. Examination of vibrio parahaemolyticus
ICS 07.100.30
C53
National Standards of People's Republic of China
Replacing GB/T 4789.7-2003
Microbiological examination of food hygiene
Vibrio parahaemolyticus test
Posted 2008-05-16
2008-11-01 implementation
People's Republic of China Ministry of Health
Standardization Administration of China released
Foreword
The revised standard adopted by the US Food and Drug Administration (FDA) "Bacteriological Analytical Manual," Chapter 9. Cholera Vibrio parahaemolyticus arc
The main difference between standard and FDA method compared as follows.
--- The sample preparation sample size 50g (mL) is amended as 25g (mL);
--- Bacteria will increase time 16h ~ 18h modify 8h ~ 18h;
--- Increase CHROMagar Vibrio selective plates;
--- Automatic increase bacterial biochemical identification system VITEK;
--- Epitope added a new serotype.
This standard replaces GB/T 4789.7-2003 "Microbiological examination of food hygiene Vibrio parahaemolyticus test." Since the implementation of this standard
Date, GB/T 4789.7-2003 repealed simultaneously.
This standard compared with GB/T 4789.7-2003 main changes are as follows.
--- Selective enrichment broth of sodium chloride crystal violet enrichment broth to 3% sodium chloride aqueous alkaline peptone;
--- The selective separation medium consisting of sodium chloride, sucrose agar, and selective agar Halophiles to thiosulfate - citrate - bile -
Sucrose agar and CHROMagar vibrio chromogenic medium;
--- Increase API20E diagnostic test strips and automatic biochemical bacterial identification system VITEK;
--- Increase serotyping;
--- The animals Kanagawa test to test;
--- Most likely increase the number of keys.
This standard Annex A, Annex B normative appendix.
This standard is proposed and administered by the People's Republic of China Ministry of Health.
This standard is drafted by. China Center for Disease Control Nutrition and Food Safety.
Participated in the drafting of this standard. Fujian Provincial Disease Prevention and Control Center, CDC, Zhejiang Province, China Inspection and Quarantine Science RESEARCH
Study hospital, the Beijing Municipal Center for Disease Control and Prevention.
The main drafters of this standard. Xiumei, Chen Yan, flying horses, Chengsu Yun, Chen Yan Long, Chen Qian.
This standard replaces the standards previously issued as follows.
--- GB 4789.7-1984, GB/T 4789.7-1994, GB/T 4789.7-2003.
Microbiological examination of food hygiene
Vibrio parahaemolyticus test
1 Scope
This standard applies to fish and food poisoning Vibrio parahaemolyticus sample testing, reference can use other foods.
2 Equipment and Materials
In addition to the microbiological laboratory conventional sterilization and cultivation equipment, other equipment and materials as follows.
2.1 incubator. 36 ℃ ± 1 ℃.
2.2 Refrigerator. 2 ℃ ~ 5 ℃.
2.3 homogenizer or sterile mortar.
2.4 Balance. a sense of the amount of 0.1g.
2.5 sterile tube. 18mm × 180mm, 15mm × 100mm.
2.6 sterile pipette. 1mL (with 0.01mL scale), 10mL (with 0.1mL scale) or micro pipettes and tips.
2.7 sterile conical flask. 500mL, 250mL.
2.8 sterile Petri dish. diameter of 90mm.
2.9 automated microbial identification system (VITEK) 1).
2.10 sterile surgical scissors, tweezers.
3 media and reagents
3.1 3% sodium chloride alkaline peptone water (APW). See Section A. Chapter 1.
3.2 thiosulfate - citrate - bile - sucrose (TCBS) agar. See Section A. 2.
3.3 3% sodium chloride tryptone soy (TSA) agar. See Section A. 3.
3.4 3% sodium chloride triple sugar iron (TSI) agar. See Section A. 4.
3.5 halophilic test medium. see Section A. 5.
3.6 3% sodium chloride mannitol test medium. see Section A. 6.
3.7 3% sodium chloride lysine decarboxylase test medium. see Section A. 7.
3.8 3% sodium chloride MR-VP Medium. see Section A. 8.
3.9 My wife's blood agar. see Section A. 9.
3.10 oxidase reagents. see Section A. Chapter 10.
3.11 Gram stain solution. See Section A. 11.
3.12 ONPG reagent. see Section A. Chapter 12.
3.13 Voges-Proskauer (V-P) Reagents. see Section A. 13.
1) Product name supplied by the French company bioMérieux products. This information is given for the convenience of users of this standard does not mean that the product
Recognition. If other equivalent product has the same effect, you can use these equivalent products.
2) trade names provided by the French CHROMagar company's products. This information is given for the convenience of users of this standard does not mean that the product identified
can. If other equivalent product has the same effect, you can use these equivalent products.
3.14 CHROMagar (CHROMagar) vibrio chromogenic medium 2).
3.15 API20E biochemical identification kits and biochemical identification card or VITEKNFC 1).
......
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