TECHNICAL DATA SHEET
TRYPTO-CASEIN SOY AGAR HALAL (TSA HALAL)
NUTRIENT MEDIA
1 INTENDED USE
Trypto Casein-Soy Agar is a universal nutrient medium suitable for a wide range of uses. In light of its excellent nutritive
value, it can be used for the growth and isolation of both aerobic and anaerobic bacteria and to favor the development
of the most fastidious microorganisms. Poured in plates or on strips, it is useful in rapid tests for examining the
contamination of surfaces. In addition, under the denomination TSA (Tryptic Soy Agar), this formula corresponds to the
reference medium used for the evaluation of productivity and selectivity criteria in the context of the ISO 11133 standard.
The typical composition corresponds to that defined in European (EP), United States (UP) and Japanese (JP)
Pharmacopeias.
The medium is certified Halal by Halal Feed and Food Inspection Authority (HFFIA), The Hague, Netherlands.
2 PRINCIPLES
The combination of enzymatic digest of casein and Papaic digest of soybean meal, both Halal certified, leads to optimal
growth, due to the synergy between the protein supply of casein and the carbohydrate supply of soybeans, favoring the
growth of most fastidious or non-fastidious microorganisms.
Sodium chloride maintains osmotic balance.
3 TYPICAL COMPOSITION
The composition can be adjusted in order to obtain optimal performance.
For 1 liter of media:
- Enzymatic digest of Casein ........................................................................................ 15.0 g
- Papaic digest of soybean meal .................................................................................... 5.0 g
- Sodium chloride ........................................................................................................... 5.0 g
- Bacteriological agar.................................................................................................... 15.0 g
pH of the ready-to-use media at 25 °C: 7.3 ± 0.2.
4 PREPARATION
Preparation from dehydrated medium:
• Dissolve 40.0 g of dehydrated medium (BK244) in 1 liter of distilled or demineralized Reconstitution:
water. 40.0 g/L
• Slowly bring to boiling, stirring with constant agitation until complete dissolution.
• Dispense in tubes or flasks. Sterilization:
• Sterilize in an autoclave at 121°C for 15 minutes. 15 min at 121 °C
• Cool and maintain the media in a molten state at 44-47 °C.
Rue des Quarante Mines – ZAC de Ther – Allonne – B.P. 10245 - 60002 Beauvais Cedex – France Page 1 sur 4
Tél : + 33 (0)3 44 14 33 33 – Fax : + 33 (0)3 44 14 33 34 - [Link]/biokar-diagnostics/
Use of ready-to-melt media:
• Using the media prepared in advance as above, melt the agar for the minimum amount of time necessary to
achieve total liquefaction.
• Cool and maintain the media in a molten state at 44-47 °C.
5 INSTRUCTIONS FOR USE
Surface enumeration Inoculation:
• Pour the molten media held at 44-47 °C into sterile Petri plates. 0.1 mL on surface
• Let solidify on a cold, flat surface.
• Dry the plates in an incubator with the covers partially removed. Incubation:
• To the surface of plates prepared as above, inoculate 0.1 mL of the test sample 30-35 °C
and its serial with a sterile triangle or “hockey stock“.
• Incubate according to the reference applied:
- at 30-35 °C for 72 ± 6 hours (NF EN ISO 21149)
- at 30-35 °C for 48 to 72 hours (NF EN ISO 18415)
- at 30-35 °C for up to 3 days for bacteria and up to 5 days for yeasts and molds (Pharmacopeias)
NOTE:
The media can also be used after having filtered through a nitrocellulose membrane (Pharmacopeias, NF EN ISO
21149)
Enumeration in pour plates
Inoculation:
• Transfer 1 mL of the initial suspension and its serial dilutions to successive sterile 1 mL in pour plates
Petri plates.
• Pour roughly 15 mL of molten media per plate. Incubation:
• Homogenize well and let solidify on a cold, flat surface. 30-35 °C
• Incubate according to the reference applied:
- at 30-35 °C for 72 ± 6 hours (NF EN ISO 21149)
- at 30-35 °C for 48 to 72 hours (NF EN ISO 18415)
- at 30-35 °C for up to 3 days for bacteria and up to 5 days for yeasts and molds (Pharmacopeias)
For other uses, refer to the references in vigor.
6 RESULTS
Following incubation, observe bacterial growth.
When performing enumeration, only retain the plates containing between 30 and 300 colonies.
7 QUALITY CONTROL
Dehydrated media: cream-white powder, free-flowing and homogeneous.
Prepared media: amber agar.
Typical culture response after incubation at 30-35 °C, inoculum ≤ 102 microorganisms (NF EN ISO 11133; EP, USP & JP):
Microorganisms Duration Growth (Productivity Ratio: PR)
Staphylococcus aureus WDCM 00032 Positive, PR ≥ 70 %
Pseudomonas aeruginosa WDCM 00026 Positive, PR ≥ 70 %
Bacillus subtilis ssp. spizizenii WDCM 00003 20 h Positive, PR ≥ 70 %
Escherichia coli WDCM 00012 Positive, PR ≥ 70 %
Salmonella Typhimurium WDCM 00031 Positive, PR ≥ 70 %
Bacillus cereus WDCM 00001 PR ≥ 70 %
Listeria monocytogenes 4 b WDCM 00021 48H PR ≥ 70 %
Staphylococcus aureus WDCM 00034 PR ≥ 70 %
Candida albicans WDCM 00054
Up to 5 days Positive
Aspergillus brasiliensis WDCM 00053
Rue des Quarante Mines – ZAC de Ther – Allonne – B.P. 10245 - 60002 Beauvais Cedex – France Page 2 sur 4
Tél : + 33 (0)3 44 14 33 33 – Fax : + 33 (0)3 44 14 33 34 - [Link]/biokar-diagnostics/
8 STORAGE / SHELF LIFE
Dehydrated medium: 2-30 °C.
The expiration dates are indicated on the labels.
Prepared media in tubes or vials (*): 180 days at 2-25 °C.
Prepared media in plates (*): 30 days at 2-8 °C.
(*) Benchmark value determined under standard preparation conditions, following manufacturer’s instructions.
9 PACKAGING
Dehydrated medium:
500 g bottle ........................................................................................................................................................... BK244HA
5 kg drum ............................................................................................................................................................. BK244GC
10 BIBLIOGRAPHY
NF EN ISO 9308-1. September 2014. Water quality - Enumeration of Escherichia coli and coliform bacteria - Part 1 :
membrane filtration method for waters with low bacterial background flora.
NF EN ISO 9308-1/A1. Janvier 2017. Water quality - Enumeration of Escherichia coli and coliform bacteria - Part 1 :
membrane filtration method for waters with low bacterial background flora – Amendment 1.
NF EN ISO 11731. July 2017. Water quality - Enumeration of Legionella.
NF EN ISO 11133. July 2014. Microbiology of food, animal feed and water - Preparation, production, storage and
performance testing of culture media (Tirage 2 (2016-01-01)).
NF EN ISO 11133/A1. March 2018. Microbiology of food, animal feed and water - Preparation, production, storage and
performance testing of culture media – Amendment 1.
NF EN 15784. November 2021. Animal feeding stuffs: methods of sampling and analysis - Detection and enumeration
of Bacillus spp. used as feed additive.
NF EN ISO 22964. June 2017. Microbiology of the food chain - Horizontal method for the detection of Cronobacter spp.
NF EN ISO 4973. October 2023. Cosmetics - Microbiology - Quality control of culture media and diluents used in
cosmetics standards.
NF EN ISO 11930. February 2019. Cosmetics - Microbiology - Evaluation of the antimicrobial protection of a cosmetic
product.
NF EN ISO 11930/A1. September 2022. Cosmetics - Microbiology - Evaluation of the antimicrobial protection of a
cosmetic product – Amendment 1.
NF EN ISO 18415. August 2017. Cosmetics - Microbiology - Detection of specified and non-specified microorganisms.
NF EN ISO 18415/A1. September 2022. Cosmetics - Microbiology - Detection of specified and non-specified
microorganisms – Amendment 1.
NF EN ISO 21149. August 2017. Cosmetics - Microbiology - Enumeration and detection of aerobic mesophilic bacteria.
NF EN ISO 21149/A1. September 2022. Cosmetics - Microbiology - Enumeration and detection of aerobic mesophilic
bacteria – Amendment 1.
NF EN ISO 21150. February 2016. Cosmetics - Microbiology - Detection of Escherichia coli.
NF EN ISO 21150. September 2022. Cosmetics - Microbiology - Detection of Escherichia coli - Amendment 1. NF EN
ISO 22717. February 2016. Cosmetics - Microbiology - Detection of Pseudomonas aeruginosa.
NF EN ISO 22717/A1. September 2022. Cosmetics - Microbiology - Detection of Pseudomonas aeruginosa -
Amendment 1.
NF EN ISO 22718. February 2016. Cosmetics - Microbiology - Detection of Staphylococcus aureus.
NF EN ISO 22718/A1. September 2022. Cosmetics - Microbiology - Detection of Staphylococcus aureus - Amendment
1
European Pharmacopeia. Chapter 2.6.12 Microbiological examination of non-sterile products: Microbial enumeration
tests and Chapiter 2.6.13. Microbiological examination of non-sterile products: Test for specified microorganisms.
Harmonized method.
Rue des Quarante Mines – ZAC de Ther – Allonne – B.P. 10245 - 60002 Beauvais Cedex – France Page 3 sur 4
Tél : + 33 (0)3 44 14 33 33 – Fax : + 33 (0)3 44 14 33 34 - [Link]/biokar-diagnostics/
The United States Pharmacopeia. Chapter <61> Microbiological examination of non-sterile products: Microbial
enumeration tests and Chapter <62> Microbiological examination of non-sterile products: Test for specified products.
The Japanese Pharmacopoeia. Chapter 4.05 Microbial Limit Test I. Microbiological examination of non-sterile products:
Total viable aerobic count and II. Microbiological examination of non-sterile products: Test for specified products.
11 ADDITIONAL INFORMATION
The information provided on the labels take precedence over the formulations or instructions described in this
document and are susceptible to modification at any time, without warning.
Document code : TRYPTO-CASEINE-SOY-AGAR-HALAL_BK244_V2(en)
Creation date : 03-2024
Updated : 11-2024
Origin of revision : Update of quality control
Rue des Quarante Mines – ZAC de Ther – Allonne – B.P. 10245 - 60002 Beauvais Cedex – France Page 4 sur 4
Tél : + 33 (0)3 44 14 33 33 – Fax : + 33 (0)3 44 14 33 34 - [Link]/biokar-diagnostics/
