GB/T 13091-2018 PDF English
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| Standard ID | Contents [version] | USD | STEP2 | [PDF] delivery | Name of Chinese Standard | Status |
| GB/T 13091-2018 | English | 255 |
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Determination of Salmonella in feeds
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| GB/T 13091-2002 | English | 759 |
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Determination of salmonella in feeds
| Obsolete |
| GB/T 13091-1991 | English | 479 |
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Method for determination of Salmonella in feeds
| Obsolete |
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GB/T 13091-2018: Determination of Salmonella in feeds---This is an excerpt. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.), auto-downloaded/delivered in 9 seconds, can be purchased online: https://www.ChineseStandard.net/PDF.aspx/GBT13091-2018
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
ICS 65.120
B 46
Replacing GB/T 13091-2002
Determination of Salmonella in feeds
Issued on. SEPTEMBER 17, 2018
Implemented on. APRIL 01, 2019
Issued by. State Administration for Market Regulation;
Standardization Administration of PRC.
Table of Contents
Foreword... 3
1 Scope... 4
2 Normative references... 4
3 Medium or material... 4
4 Instruments and equipment... 5
5 Samples... 5
6 Test method (see Appendix A for the test procedure diagram)... 6
7 Expression of results... 10
Appendix A (Normative) Test procedure diagram... 11
Appendix B (Normative) Preparation and test methods of medium and reagents... 12
Foreword
This standard was drafted in accordance with the rules given in GB/T 1.1-2009.
This standard replaces GB/T 13091-2002 "Determination of Salmonella in feeds".
Compared with GB/T 13091-2002, except for editorial modifications, the main changes
of this standard are as follows.
- DELETE the terms and definitions (see Chapter 3 of the 2002 edition);
- DELETE the principle (see Chapter 4 of the 2002 edition);
- ADD the test procedure diagram (see Figure A.1 in Appendix A);
- MODIFY the enrichment equipment; ADD the homogenization bag and
homogenizer options (see 6.1; 9.1.2 of the 2002 edition);
- CHANGE the separation medium from "phenol red brilliant green agar and DHL
agar" to "BS agar, DHL agar or Salmonella chromogenic medium" (see 6.3; 9.4 of
the 2002 edition);
- MODIFY the identification process and identification table (see 6.4; 9.5 of the 2002
edition);
- ADD the step of "preliminary judgment using trisaccharide iron test and lysine
decarboxylase test" (see 6.4.1);
- ADD the option of "Biochemical identification kit or automatic microbial
biochemical identification system" for Salmonella identification (see 6.4.3).
This standard was proposed by AND shall be under the jurisdiction of the National Feed
Industry Standardization Technical Committee (SAC/TC 76).
Drafting organizations of this standard. Institute of Agricultural Quality Standards and
Testing Technology, Chinese Academy of Agricultural Sciences [National Feed Quality
Supervision and Inspection Center (Beijing)].
The main drafters of this standard. Rao Zhenghua, Liu Xiaolu, Fan Xia.
This standard replaces the standard previously issued as follows.
- GB/T 13091-1991, GB/T 13091-2002.
Determination of Salmonella in feeds
1 Scope
This standard specifies the inspection methods for Salmonella in feed.
This standard applies to the inspection of Salmonella in feed and feed additives.
2 Normative references
The following documents are essential to the application of this document. For the dated
documents, only the versions with the dates indicated are applicable to this document;
for the undated documents, only the latest version (including all the amendments) is
applicable to this standard.
GB/T 6682 Water for analytical laboratory use - Specification and test methods
3 Medium or material
Unless otherwise stated, only reagents confirmed to be of analytical grade are used in
the analysis.
3.1 Water. It shall meet the requirements of grade-3 water in GB/T 6682.
3.2 Buffered peptone water (BPW). See B.1 in Appendix B.
3.3 Magnesium chloride malachite green (RV) enrichment solution. See B.2 in
Appendix B.
3.4 Cystine selenite (SC) enrichment solution. See B.3 in Appendix B.
3.5 Bismuth sulfite (BS) agar. See B.4 in Appendix B.
3.6 Deoxycholate-Hydrogen-Sulfide-Lactose (DHL) agar. See B.5 in Appendix B.
3.7 Salmonella chromogenic medium.
4 Instruments and equipment
4.1 Refrigerator. 2 °C ~ 5 °C.
4.2 Constant temperature incubator. 36 °C ± 1 °C, 42 °C ± 1 °C.
4.3 Homogenizer.
4.8 Sterile petri dishes. Diameter 60 mm, 90 mm.
4.9 Sterile test tubes. 3 mm × 50 mm, 10 mm × 75 mm.
4.10 pH meter or pH colorimetric tube or precision pH test paper.
4.11 Automatic microbial biochemical identification system.
5 Samples
5.1 Sampling principles
The collection of samples shall follow the principles of randomness and
representativeness. The sampling process shall follow aseptic procedures, to prevent all
possible external contamination.
5.2 Sampling method
5.2.4 For solid products with individual packages greater than 500 g, use a sterile
sampler to take appropriate samples from different parts of the same package; put them
into the same sterile sampling container, as one sample.
5.3 Storage and transport of collected samples
5.3.1 The samples shall be sent to the laboratory for testing, as soon as possible.
6 Test method (see Appendix A for the test procedure diagram)
6.1 Pre-enrichment
Weigh 25g (mL) of sample under sterile conditions. Add it into a 500 mL sterile conical
flask, which was filled with 225 mL of sterile BPW. Place it in an oscillator. Oscillate
at 8000 r/min ~ 10000 r/min for 2 min ~ 3 min. If the sample is in liquid form, shake
and mix well. Incubate it at 36 °C ± 1 °C for 18 h ± 2 h.
6.2 Selective enrichment
After the pre-enrichment culture is shaken, take 1 mL to inoculate it into a test tube,
which contains 10 mL of RV. Incubate it at 42 °C ± 1 °C for 18 h ~ 24 h. Take another
1 mL of pre-enrichment culture. Inoculate it into a test tube, which contains 10 mL of
SC. Incubate it at 36 °C ± 1 °C for 18 h ~ 24 h.
6.3 Isolation and culture
Use an inoculation loop, to take 1 loop of selective enrichment solution. Streak and
inoculate it on a BS agar plate. Incubate it at 36 °C ± 1 °C for 40 h ~ 48 h. Take another
loop of selective enrichment solution. Streak and inoculate it on a DHL agar plate or a
Salmonella chromogenic medium plate.
6.4 Biochemical test
6.4.1 Pick 2 or more typical or suspicious colonies from the selective agar plate.
Inoculate on trisaccharide iron agar. First streak the slope. Then puncture the bottom
layer. Do not sterilize the inoculation needle. Directly inoculate lysine decarboxylase
test medium and nutrient agar plate. Culture at 36 °C ± 1 °C for 18 h ~ 24 h. If necessary,
it can be extended to 48 h. The changes in the characteristics of the trisaccharide iron
agar medium are as shown in Table 2.Table 3 shows the reaction results of Salmonella
in the trisaccharide iron agar and lysine decarboxylase test medium.
6.5 Serological identification
6.5.1 Checking the culture for autoagglutination
Use 1.2% ~ 1.5% agar culture as the antigen, for slide agglutination test. Firstly, to rule
out self-agglutination reaction, add a drop of normal saline to a clean glass slide; mix
the culture to be tested in the normal saline, to make a homogeneous turbid suspension;
shake the slide gently for 30 s ~ 60 s; observe the reaction in dark (observe with a
magnifying glass if necessary). If there is visible bacterial agglutination, it is considered
to have self-agglutination; otherwise, there is no self-agglutination. For the non-self-
agglutinating culture, refer to the following method for serological identification.
6.5.2 Identification of the O antigen
Draw 2 areas of about 1 cm × 2 cm on the slide. Pick 1 loop of bacteria to be tested. Put
1/2 loop on the upper part of each area on the slide. Add 1 drop of multivalent bacteria
O serum, to the lower part of one of the areas.
6.5.3 Identification of H antigen
The operation is the same as 6.5.2.When the H antigen is underdeveloped, inoculate
the strain in the center of a 0.55% ~ 0.65% semi-solid agar plate. When the colony
spreads and grows, take bacteria from the edge for inspection. OR inoculate the strain
1 ~ 2 times in small glass tubes, which contain 0.3% ~ 0.4% semi-solid agar plate. Take
the bacteria from the far end for culture and re-inspection.
6.5.4 Identification of Vi antigen
The operation is the same as 6.5.2.Check with Vi factor serum.
7 Expression of results
Based on the results of the above biochemical tests and serological tests, it is concluded
that Salmonella is detected or not detected in 25g (mL) samples.
......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
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