GB/T 18204.3-2013 (GB/T18204.3-2013, GBT 18204.3-2013, GBT18204.3-2013) & related versions
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Examination methods for public places -- Part 3: Airborne microorganism
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GB/T 18204.3-2013
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GBT 18204.3-2013
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Methods of microbiological examination for tea set in public places. Determination of coliform bacteria
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GB/T 18204.3-2013
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
ICS 13.060
C 51
Replacing GB/T 18204.1-2000
Partially replacing GB/T 17220-1998
Examination methods for public places –
Part 3. Airborne microorganism
ISSUED ON. DECEMBER 31, 2013
IMPLEMENTED ON. DECEMBER 1, 2014
Issued by. General Administration of Quality Supervision, Inspection and
Quarantine;
Standardization Administration Committee.
Table of Contents
Foreword ... 3
1 Scope ... 5
2 Terms and definitions ... 5
3 Total bacterial count ... 6
4 Total fungi count ... 8
5 β-hemolyticstreptococcus ... 9
6 Legionella pneumophila ... 10
Annex A (normative) Requirements for site sampling point arrangement ... 14
Foreword
GB/T 18204 "Examination methods for public places" consists of the following
6 parts.
- Part 1. Physical parameters;
- Part 2. Chemical pollutants;
- Part 3. Airborne Microorganism;
- Part 4. Microorganism on a surface of public articles;
- Part 5. Central Air Conditioning Ventilation System;
- Part 6. Technical specifications of health monitoring.
This Part is Part 3 of GB/T 18204.
This Part was drafted in accordance with the rules given in GB/T 1.1-2009.
This Part replaces GB/T 18204.1-2000 "Methods of microbiological
examination for air in public places - Determination of aerobic bacterial count",
partially replaces GB/T 17220-1998 "Technical rules of health monitoring for
public places".
Compared with GB/T 18204.1-2000 and GB/T 17220-1998, the main changes
in this Standard are as follows.
- added the inspection method for total fungi count;
- added the inspection method for β-hemolytic streptococcus;
- added the inspection method for legionella pneumophila.
This Part was proposed by and shall be under the jurisdiction of Ministry of
Health of the People's Republic of China.
This Part shall be interpreted by Ministry of Health of the People's Republic of
China.
Main drafting organizations of this Part. China Center for Disease Control and
Prevention Center for Environment and Health-related Product Safety.
The drafting organizations of this Part. Jiangsu Provincial Center for Disease
Control and Prevention, Shenzhen Center for Disease Control and Prevention,
Ma'anshan City Health Bureau Health Supervision Office.
Examination methods for public places -
Part 3. Airborne microorganism
1 Scope
This Part of GB/T 18204 specifies the field sampling and laboratory culture
methods for total bacterial count, total fungi count, β-hemolytic streptococcus
and legionella pneumophila in public air.
This Part is applicable to the determinations of total bacterial count, total fungi
count, β-hemolytic streptococcus and legionella pneumophila in public air.
Other places can refer to the implementation.
NOTE If there are 2 or more inspection methods for the same indicator in this Part, they can
be selected according to the technical conditions.
2 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
2.1 total bacterial count
the total number of mesophilic aerobic and facultative anaerobic colonies of
samples collected in public air counted on nutrient agar medium that are grown
and cultured at 35°C~37°C for 48h
2.2 total fungi count
the number of colonies of samples collected in public air counted on Sabouraud
agar that are cultured at 28°C for 5d
2.3 β-hemolytic streptococcus
typical colonies formed by samples collected in public air on blood agar dish at
35°C~37°C for 24h ~ 48h
2.4 legionella pneumophila
colonies that samples are grown on GVPC agar dish to generate typical
colonies and grown on BCYE agar dish but not grown on L-cysteine-depleted
BCYE agar dish and further confirmed by biochemical and serological assays
Sodium chloride 5 g
Agar 20 g
Defibrinated sheep blood 5mL ~ 10mL
Distilled water 1000 mL
5.2.2.2 Preparation method. dissolve peptone, sodium chloride and meat
paste in distilled water, calibration pH is 7.4~7.6, add agar, sterilize at 121°C for
20min. After cooling to about 50°C, add defibrinated sheep's blood aseptically,
and shake the dish.
5.2.3 Sampling
See 3.2.3.
5.2.4 Inspection steps
5.2.4.1 Cultivation method. the sampled blood agar dishes are cultured at
35°C~37°C for 24h~48h.
5.2.4.2 Results observation. after the cultivation, fine colonies appearing on
the blood agar dish are grayish white and protrude from the surface and have
a diameter of 0.5mm ~ 0.7mm. Colonies are transparent or translucent, with a
smooth surface. Microscopic examination is gram-positive bacillus-free, round
or oval, arranged in a chain. The length of the chain varies from 4 to 8 cells to
several tens of cells depending on the culture and operating conditions. There
is a clear, completely transparent and colorless hemolytic ring with 2mm~4mm
around the colonies. The colonies that meet the above characteristics shall be
β-hemolytic streptococcus.
5.2.5 Results report
5.2.5.1 Calculation of β-hemolytic streptococcus of sampling point. count the
colonies, record the results and convert to CFU/m3 (colony forming units per
cubic meter of air) based on the dilution ratio and volume of the produced gas.
5.2.5.2 Determination results of β-hemolytic streptococcus of one area. the
determination of β-hemolytic streptococcus in the air in one area is given as the
maximum of the β-hemolytic streptococcus in all sampling points in the area.
6 Legionella pneumophila
6.1 Principle
Use liquid impacting method to sample, cultivation method to determine
legionella pneumophila in the air in public places.
6.2 Instruments and equipment
6.3.2 Sampling absorbent 2 - yeast extract
6.3.2.1 Absorbent composition.
Yeast extract powder 12 g
Distilled water 1000 mL
6.3.2.2 Preparation method. add yeast extract powder with water to 1000 mL,
sterilize at 121°C for 15min, dispense in sterile centrifuge tubes (6.2.3) for use.
6.3.3 Potassium chloride hydrochloride solution [c(HCl·KCl)=0.01 mol/L]
6.3.3.1 Composition.
Hydrochloric acid (0.2 mol/L) 3.9 mL
Potassium chloride (0.2 mol/L) 25 mL
6.3.3.2 Preparation method. mix the above ingredients, adjust pH=2.2±0.2
with 1 mol/L potassium hydroxide, sterilize at 121°C for 15min for use.
6.3.4 GVPC agar plate.
6.3.5 BCYE agar plate.
6.3.6 BCYE-CYE agar plate.
6.3.7 Gram staining solution.
6.3.8 Urate biochemical reaction tube.
6.3.9 Legionella typing serum reagents.
6.4 Sampling
6.4.1 Sampling point. see Annex A.
6.4.2 Pour 20 mL of sampling absorbent 1 (6.3.1) into the microorganism
aerosol sampler (6.2.2), then add 1 to 2 drops of mineral oil with a straw.
6.4.3 Connect the microorganism aerosol concentrator (6.2.1) and the liquid
impact microorganism aerosol sampler (6.2.2). Adjust the main flow and
concentrated flow according to the flow requirements for the microorganism
aerosol concentrator and the liquid impact microorganism aerosol sampler.
6.4.4 According to the instructions of the concentrator and sampler, the
amount of collected air for each aerosol sample is 1m3 ~ 2m3.
6.4.5 Pour 20mL of sample absorbent 2 (6.3.2) into the liquid impact
microorganism aerosol sampler (6.2.2). Then add 1 to 2 drops of mineral oil
with a straw. Repeat 6.4.3, 6.4.4 steps at the same sampling point.
Annex A
(Normative)
Requirements for site sampling point arrangement
A.1 Scope
This appendix specifies the basic requirements for the site sampling point
arrangement for microbiological impacting method and natural sinking method
in air in public places.
A.2 Site sampling point arrangement of impacting method
A.2.1 Set one sampling point when the indoor area is less than 50 m2, two
sampling points for indoor area of 50m2 ~ 200m2, three to five sampling points
for indoor area over 200m2.
A.2.2 Sampling points are arranged according to the principle of even
distribution. The one sampling point in the room is set in the center. The two
sampling points are set on the indoor symmetrical point. The three sampling
points are set on the three equally divided points of the indoor diagonal. The
five sampling points are arranged according to plum blossom points. Others are
laid out according to the principle of even distribution.
A.2.3 The sampling point is 1.2m~1.5m above the ground and no less than
1m from the wall.
A.2.4 The sampling points shall avoid ventilation openings, ventilation
passages, etc.
A.3 Site sampling point arrangement of natural sinking method
A.3.1 Set three sampling points when the indoor area is less than 50 m2, five
sampling points for indoor area over 50m2.
A.3.2 Sampling points are arranged according to the principle of even
distribution. The three sampling points in the room are set on three equal points
on the indoor diagonal. The five sampling points are arranged according to plum
blossom points.
A.3.3 The sampling point is 1.2m~1.5m above the ground and no less than
1m from the wall.
A.3.4 The sampling points shall avoid ventilation openings, ventilation
passages, etc.
......
GB/T 18204.3-2000
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
ICS 13.020
C 51
Methods of microbiological examination for tea set in
public places - Determination of Coliform bacteria
ISSUED ON: SEPTEMBER 30, 2000
IMPLEMENTED ON: JANUARY 01, 2001
Issued by: State Bureau of Quality Technical Supervision
Abolished (Replaced by GB/T 18204.4-2013)
Table of Contents
Foreword ... 3
1 Scope ... 4
2 Definition ... 4
3 Instruments ... 4
4 Medium and reagents ... 5
5 Testing procedures ... 7
6 Operation steps ... 8
7 Report of results ... 9
Methods of microbiological examination for tea set in
public places - Determination of Coliform bacteria
1 Scope
This standard specifies the method for determination of coliform bacteria in tea
sets in public places.
This standard applies to the determination of coliform bacteria in public tea sets
in public places.
2 Definition
This standard uses the following definition.
Coliform bacteria
Refers to a group of Gram-negative non-bacillus bacteria, that can ferment
lactose, produce acid, produce gas, aerobic and facultative anaerobic, when
cultured at 37 °C for 24 hours.
The bacteria are mainly derived from human and animal feces, so it is used as
an indicator of fecal pollution, to evaluate the sanitary quality of the test object.
3 Instruments
3.1 High-pressure steam sterilizer.
3.2 Dry heat sterilization box.
3.3 Incubator: 36 °C ± 1 °C.
3.4 Water tank.
3.5 Electric stove.
3.6 Balance.
3.7 Sterilized plate: 9 cm in diameter.
3.8 Sterilized graduated straws: 1 mL, 10 mL.
3.9 Sterilized cotton swabs.
3.10 Sterilized scissors.
3.11 pH meter or precision pH test paper.
4 Medium and reagents
4.1 Lactose bile salt fermentation broth
4.1.1 Ingredients:
Peptone: 20 g
Porcine bile salt (or beef and sheep bile salt): 5 g
Lactose: 10 g
0.04% bromocresol purple aqueous solution: 25 mL
Distilled water: 1000 mL
4.1.2 Preparation method: Dissolve peptone, bile salt, lactose in distilled water.
Adjust the pH to 7.4. Add bromocresol purple solution. Mix well. Dispense into
test tubes, which are equipped with inverted tubes, 10 mL per tube. Autoclave
at 115 °C for 15 min.
Note: The double-material lactose bile salt fermentation tube has doubled other
ingredients. except distilled water.
4.2 Eosin methylene blue agar
4.2.1 Ingredients:
Egg white: 10 g
Lactose: 10 g
Dipotassium hydrogen phosphate: 2 g
Agar: 17g
2% eosin aqueous solution: 20 mL
0.65% methylene blue solution: 10 mL
Distilled water: 1000 mL
4.2.2 Preparation method: Dissolve peptone, phosphate, agar in distilled water.
4.4.4 Safranine counterstaining solution:
Safranine: 0.25 g
95% ethanol: 10 mL
Distilled water: 90 mL
Dissolve safranine in ethanol. Then use distilled water to dilute it.
4.5 Staining method
4.5.1 Fix the smear on the flame. Add crystal violet staining solution dropwise.
Stain for 1 min. Use water to wash it.
4.5.2 Add gram iodine solution dropwise. Let it act for 1 min. Use water to wash
it.
4.5.3 Add 95% ethanol dropwise to decolorize, for about 30 s. Use water to
wash it.
4.5.4 Add counterstain solution dropwise, to counterstain for 1 min. Use water
to wash it. Wait for it to dry. Check with microscope.
4.6 Staining results
Gram-positive bacteria are purple; gram-negative bacteria are red.
4.7 Rapid test paper for coliform bacteria
4.7.1 Quality requirements: The paper must meet the quality standards for fast
inspection of coliform bacteria for food (drinking) utensils.
4.7.2 Quality standard: Each piece of paper is 5 cm x 5 cm; the pH value is 7.0
~ 7.4; the packaging is sterile. The manufacturer has been reviewed and
approved by the health department.
5 Testing procedures
The testing procedure for coliform bacteria is as follows:
......
| Standard ID | GB/T 18204.3-2013 (GB/T18204.3-2013) | | Description (Translated English) | Examination methods for public places. Part 3: Airborne microorganism | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | C51 | | Classification of International Standard | 13.060 | | Word Count Estimation | 12,126 | | Older Standard (superseded by this standard) | GB/T 18204.1-2000; GB/T 17220-1998 | | Drafting Organization | China Disease Prevention and Control Center Environmental Health and Related Product Safety | | Administrative Organization | People's Republic of China Ministry of Health | | Regulation (derived from) | National Standards Bulletin 2013 No. 27 | | Proposing organization | Ministry of Health of the People's Republic of China | | Issuing agency(ies) | General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China, Standardization Administration of the People's Republic of China | | Summary | This standard specifies the method of field sampling and laboratory cultured air of public places total bacteria, total fungi, ��-hemolytic streptococci and Legionella pneumophila bacteria. This standard applies to the total number of public places, airbor | | Standard ID | GB/T 18204.3-2000 (GB/T18204.3-2000) | | Description (Translated English) | Methods of microbiological examination for tea set in public places. Determination of coliform bacteria | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | C51 | | Classification of International Standard | 13.020 | | Word Count Estimation | 5,585 | | Date of Issue | 2000-09-30 | | Date of Implementation | 2001-01-01 | | Drafting Organization | Health and Epidemic Prevention Station of Jiangsu Province | | Administrative Organization | Chinese Academy of Preventive Medicine | | Regulation (derived from) | National Standards Bulletin 2013 No. 27 | | Proposing organization | Ministry of Health of the People Republic of China | | Issuing agency(ies) | State Quality and Technical Supervision | | Summary | This standard specifies: Tea coliforms in public places determination. This standard applies to: Public tea coliforms in public areas determination. |
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