This standard specifies the test method for Vibrio parahaemolyticus. This standard is applicable to the test of Vibrio parahaemolyticus in dried aquatic animal products, pickled and raw aquatic animal products, ready-to-eat algae products, surimi products and other aquatic foods and aquatic condiments as well as poisoning samples. GB/T 4789.7-2003 Food Hygiene Microbiological Test Vibrio parahaemolyticus Test GB/T4789.7-2003 Standard Download Decompression Password: www.bzxz.net

ICS07.100.30
National Standard of the People's Republic of China
GB/T4789.7—2003
Replaces GB/T4789.7—1994
Microbiological examination of food hygiene—Examination of Vibrio parahaemolyticus
Microbiological examination of food hygiene—Examination of Vibrio parahaemolyticusIssued on August 11, 2003
Ministry of Health of the People's Republic of China
Standardization Administration of the People's Republic of China
Implementation on January 1, 2004
GB/T4789.7-2003
This standard revise GB/T4789.7-1994 “Microbiological examination of food hygiene—Examination of Vibrio parahaemolyticus”. Compared with GB/T4789.7--1994, this standard is mainly modified as follows: the standard text format and text are modified in accordance with GB/T1.1-2000. The "equipment and materials" in the original standard are modified and standardized. The scope of application of this standard is modified to "this standard is applicable to the inspection of Vibrio parahaemolyticus in dried animal aquatic products, pickled and drunken raw animal aquatic products, ready-to-eat algae foods, fish paste products and other aquatic foods and aquatic condiments as well as food poisoning samples". From the date of implementation of this standard, GB/T4789.7-1994 will be abolished at the same time. This standard is proposed and managed by the Ministry of Health of the People's Republic of China. The drafting unit of this standard: Nutrition and Food Safety Institute of Chinese Center for Disease Control and Prevention. The main drafters of this standard: Zhou Guilian, Fu Ping, Yang Baolan. This standard was first issued in 1984 and revised for the first time in 1994. This is the second revision. 48
1 Scope
Microbiological examination of food hygiene
Vibrio parahaemolyticus examination
This standard specifies the examination method for Vibrio parahaemolyticus. GB/T4789.7—2003
This standard applies to the examination of Vibrio parahaemolyticus in dried aquatic animal products, pickled and drunken raw aquatic animal products, ready-to-eat algae foods, surimi products and other aquatic foods and aquatic condiments as well as food poisoning samples. 2 Normative references
The clauses in the following documents become the clauses of this standard through reference in this standard. For any dated referenced document, all subsequent amendments (excluding errata) or revisions are not applicable to this standard. However, the parties who reach an agreement based on this standard are encouraged to study whether the latest versions of these documents can be used. For any undated referenced document, the latest version shall apply to this standard. GB/T4789.28—2003 Food hygiene microbiology test staining method, culture medium and reagents 3 Equipment and materials
3.1 Refrigerator: 0℃~4℃.
3.2 Constant temperature incubator: 36℃±1℃.
3.3 Microscope: 10×~100×.
3.4 ​​Homogenizer or sterile mortar.
3.5 Rack-plate drug balance: 0g~500g, accurate to 0.5g. 3.6 Sterile test tube: 16mm×160mm, 15mm×100mm and corresponding test tube rack. Sterile graduated pipette: 1mL (with 0.01mL scale), 5mL, 10mL (with 0.1mL scale). 3.7
Sterile conical flask: 500mL, 200mL.
Sterile glass beads: about 5mm in diameter.
Sterile culture dish: 90mm.
1 mL syringe.
Inoculation lamp and inoculation loop.
Sterile surgical scissors, forceps, etc.
3.14 White mice: 12g~15g.
4 Culture medium and reagents
4.1 Sodium chloride crystal violet enrichment solution: in accordance with 4.39 of GB/T4789.28-2003. 4.2 Sodium chloride sucrose agar: in accordance with 4.40 of GB/T4789.28-2003. 4.3 Halophilic bacteria selective agar: in accordance with 4.41 of GB/T4789.28--2003. 4.43.5% sodium fluoride trisaccharide iron agar: in accordance with 4.42 of GB/T4789.28-2003. 4.5 Sodium chloride blood agar: in accordance with 4.43 of GB/T4789.28-2003. 4.6 Halophilic test medium: in accordance with 4.44 of GB/T4789.28-2003. 4.7 3.5% sodium fluoride biochemical test medium: in accordance with 4.45 of GB/T4789.28-2003. 4.8 Gram staining solution: in accordance with 2.2 of GB/T4789.28--2003. 49
GB/T4789.7--2003
4.9 Indigo red reagent: in accordance with 3.4 of GB/T4789.28-2003. 4.10 Indigo matrix reagent: in accordance with 3.13 of GB/T4789.28-2003. 4.11 VP reagent: in accordance with 3.4 of GB/T4789.282003. 5 Test procedure
See Figure 1 for the test procedure for Vibrio parahaemolyticus.
Sample 25g (mL) +
225mL 3.5% sterile saline, crushed
Sodium oxide crystal violet
Enrichment solution 100mL
Smear Gram staining microscopic examination of morphology
6 Operation steps
Bh~16h
Biochemical test
Animal test
Sodium chloride sucrose agar plate
And selective agar plate
37C, 18h~24h
3.5% sodium hydride tri-powder iron Material surface
37℃18h~24h
Slightly salty test
37℃, 24h
6.1 Separation and culture Take 25g (mL) of the test sample with aseptic operation, add 225mL3.5% sterile saline, break it with a homogenizer or grind it with a mortar, inoculate one sodium chloride agar plate and one halophilic bacteria selective agar plate, and take 10mL at the same time, add it to 100mL of enrichment solution, incubate it at 37℃ for 8h~16h, apply it to the above plate for separation, and incubate it at 37℃ for 18h~24h before taking it out for observation. 6.2 Pick the above suspicious colonies, transfer them to 3.5% sodium chloride trisaccharide iron slant, and observe the results at 37℃ for 24h. 6.3 Smear microscopy: For suspicious reactions of the three-sugar iron culture medium (the bottom layer turns yellow, glucose produces acid but not gas, the inclined surface does not change, lactose and sucrose do not decompose, there is power, and no hydrogen sulfide is produced), perform smear Gram staining microscopy for morphology. 6.4 Halophilicity test: Inoculate the above suspicious cultures in salt water of different concentrations, observe the growth after culturing at 37℃ for 24 hours, and conduct the following related tests for those that do not grow in salt-free or salt-containing water with more than 10% salt, and grow well in 7% salt-containing water. 6.5 Biochemical test: Inoculate various biochemical culture media respectively, place at 37℃, and observe within 24 hours except for the VP, indigo matrix, and methyl red tests, which are cultured for 48 hours and then added with reagents for observation. See Table 1 and Table 2 for specific contents. 50
Glucose acid production
Glucose gas production
Mannitol
Hydrogen sulfide
Methyl red
Table 1 Biochemical characteristics of Vibrio parahaemolyticus
Note: Ten positive; one negative ten/one majority positive, a few negative. VP
Indigo
Lysine
Ornithine
Arginine
Table 2 Main characteristics of Vibrio parahaemolyticus and identification with other Vibrio species
V.parahaemolyticus
V.alginolyticus
O, Vibrio cholerae
V. cholerae non O.
Vibrio mimicus
V. mimicus
V. fluialis
V. vulnificus
V. metschnikouii
V. hollisae
V. damsela
Glucose gas production
Lysine
Cut matrix
Ornithine
Gum arabic
GB/T4789.7-2003
B Galactose #
Salt-free waterwww.bzxz.net
Salt-aged water/(%)
Salt-free water
Note: +90% positive; -90% negative: d11%~89% positive; (+) delayed positive: (d) 11%~89% delayed positive. 7 Animal test
Inoculate the above-mentioned Vibrio parahaemolyticus with 3.5% sodium chloride aged water, culture at 37℃ for 16h~18h, inject 0.3mL into the mouse peritoneum, observe for 2d~3d, and dissect the dead mice for isolation and culture. 51
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