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Biolife Italiana Srl
Technical Sheet
N °5121583 BE –1(P) 09/2012 page 1/ 2 TSC AGAR BASE
D-CYCLOSERINE ANTIMICROBIC SUPPLEMENT
D-CYCLOSERINE 4-MUP SUPPLEMENT
Basal medium and selective supplements for the enumeration of C.perfringens in foodstuffs
according to ISO 7973 and ISO 15213

TYPICAL FORMULAS

TSC Agar Base (g/L)

D-Cycloserine Antimicrobic Supplement (vial contents for 500 mL of medium)

D-Cycloserine 4-MUP Supplement (vial contents for 500 mL of medium)

DIRECTIONS FOR POOWDERED MEDIUM
- TSC Agar (ISO 7397) : suspend 21g of TSC Agar Base in 500 mL of cold distil ed water, heat to
boiling and autoclave at 121°C for 15 minutes. Add the contents of one vial of D-Cycloserine Antimicrobic Supplement. (REF 4240002) reconstituted with 5mL of sterile distil ed water. Mix wel and pour into sterile Petri dishes. Final pH 7,6± 0,2
- TSC 4 MUP Agar: Suspend 21g of TSC Agar Base in 500mL of cold distil ed water, heat to boiling
and autoclave at 121°C for 15 minutes. Add the contents of one vial of D-Cycloserine 4-MUP Supplement. (REF 4240049) reconstituted with 5mL of sterile distil ed water. Mix wel and pour into sterile Petri dishes. Final pH 7,6± 0,2
DIRECTIONS FOR READY TO USE MEDIUM IN FLASKS
Heat to boiling the ready to use medium in flasks and cool to 45-50°C. If necessary add the suitable supplement under aseptic conditions. Mix and distribute into sterile Petri dishes.
DESCRIPTION
C. perfringens causes gastro enteric disorders characterised by abdominal pain and diarrhoea general y without fever or vomiting. The il ness is usual y proved by ingestion of inadequately cooled or reheated meats, such as meat pies, stews, gravies, etc, contaminated by soil or faeces. The food poisoning can be diagnosed using quantitative anaerobic cultures to test foods and faeces. The minimum infective dose is 105 cel s/g of food. The complete medium TSC Agar, prepared without Egg Yolk Emulsion, meets the requirements given The medium base, prepared without supplements, corresponds to “Iron Sulphite Agar” prepared
TECHNIQUE
For the enumeration of C. perfringens, ISO 7937 recommends the fol owing technique 1. Prepare the test sample, the initial suspension and the dilutions, in accordance with the specific International Standard dealing with the product concerning. ISO 6887 recommends the use of peptone salt (see Maximum Recovery Diluent REF 401691) as general diluent for foods and animal feeding Biolife Italiana S.r.l. Viale Monza 272, 20128 Milan, Italy. Tel. n° ++39 (0)2 25 209 1, Fax n° ++39 (0)2 25 76 428
E-mail: biolife@masciabrunelli.it ; Web: www.biolifeitaliana.it Biolife Italiana Srl
Technical Sheet
N °5121583 BE –1(P) 09/2012 page 2/ 2 2. Transfer by means of sterile pipettes 1 mL of the test sample (if liquid) or 1 mL of the initial suspension and 1mL of each decimal dilution, in duplicate, to the centres of empty Petri dishes. 3. Pour 15 – 20mL of Egg Yolk free TSC Agar into each dish and mix wel with the inoculum. 4. When the medium has solidified add an over layer of 10mL of the same TSC Agar. 5. Al ow to solidify and incubate in anaerobic jars or other suitable containers and incubate at 37 °C for 20 hours. Longer incubation may result in excess blackening along the bottom rim of the plates. 6. Count the black colonies on the plates containing between 15 and 150 characteristic colonies. If parts of the plates are completely blackened count the colonies at the next higher dilution even their To confirm the presence of C. perfringens the fol owing tests are recommended: For the enumeration of C. perfringens, with TSC 4 MUP Agar the fol owing technique is recommended 1. Transfer by means of sterile pipettes 0,1 mL of the test sample (if liquid) or 0,1 mL of the initial suspension and 0,1mL of each decimal dilution, in duplicate, to the surface of the TSC 4 MUP Agar 2. Incubate in anaerobic jars or other suitable containers and incubate at 44 °C for 22 ± 2 hours. 3. Count the fluorescent colonies observed under Wood’s lamp (360 nm) on the plates containing between 15 and 150 characteristic colonies. 4. Confirm the suspected colonies with the catalase test (-) and with inverted CAMP Test (+).
USER QUALITY ASSURANCE (37 °C - 24 hrs - Anaer.)
With Antimicrobial Supplement: C.perfringens ATCC 13124: growth, black green colonies (fluorescent Without Antimicrobial Supplement: C.sporogenes ATCC 19404: growth, black green colonies With Antimicrobial Supplement : E.coli ATCC 25922 : inhibited Without Antimicrobial Supplement : E.coli ATCC 25922 : growth, white colonies User prepared medium base in flasks: 2 weeks at 2-8 °C
REFERENCES
• ISO 7937 Microbiology – General guidance for enumeration of Clostridium perfringens- Colony count technique. • ISO 15213 Microbiology of food and animal feeding stuffs -- Horizontal method for the enumeration of sulfite- reducing bacteria growing under anaerobic conditions • Haushild, A.H.W. & Hilaheimer, A. (1974). App. Microbiol. 27, 78
• Harmon, S.M., Kautter, O.A. & Peeler, J.T. (1971). App. Microbiol., 22,688
• Manuel suisse des denrées alimentaires (MSDA). Chapitre 56, Microbiologie. Juil et 2000. • Shehidi, SA. & Ferguson, AR. (1971). App. Microbiol., 21, 500-606

PACKAGING
4021582 TSC Agar Base,
500g (11.9L)
4240002 D-Cycloserine Antimicrobic Suppl. 10 vials, each vial for 500 mL of basal medium.
4240049 D-Cycloserine 4-MUP Supplement,
10 vials, each vial for 500 mL of basal medium.
5121583 TSC Agar Base,
6 x 200 mL ready to use flasks
Biolife Italiana S.r.l. Viale Monza 272, 20128 Milan, Italy. Tel. n° ++39 (0)2 25 209 1, Fax n° ++39 (0)2 25 76 428
E-mail: biolife@masciabrunelli.it ; Web: www.biolifeitaliana.it

Source: http://biolifeitaliana.it/biolife/upload/file/Schede/TS-5121583.pdf