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GB/T 18204.3-2013 English PDF (GBT18204.3-2013)

GB/T 18204.3-2013 English PDF (GBT18204.3-2013)

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GB/T 18204.3-2013: Examination methods for public places -- Part 3: Airborne microorganism

This Part of GB/T 18204 specifies the field sampling and laboratory culture methods for total bacterial count, total fungi count, hemolytic streptococcus and legionella pneumophila in public air. This Part is applicable to the determinations of total bacterial count, total fungi count, hemolytic streptococcus and legionella pneumophila in public air. Other places can refer to the implementation.
GB/T 18204.3-2013
GB
NATIONAL STANDARD OF THE
PEOPLE 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 ??-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 ??-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, ??-hemolytic streptococcus and legionella pneumophila in public air.
This Part is applicable to the determinations of total bacterial count, total fungi count, ??-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 ??-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 ??-hemolytic streptococcus.
5.2.5 Results report
5.2.5.1 Calculation of ??-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 ??-hemolytic streptococcus of one area. the determination of ??-hemolytic streptococcus in the air in one area is given as the maximum of the ??-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.

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