SPS Agar

EMD Cat. No. 1.10235.0500
500 g


Medium proposed by ANGELOTTI et al. (1962) for the isolation and enumeration of Clostridium perfringens and Clostridium botulinum in all types of foodstuffs.


Additives
EMD Cat. No. Product Pack Size
1.10641.0007 Anaerocult® A mini 1 x 25
13674 Plate basket 1ea
PX0046-1 Paraffin viscous 1 l
Anaerocult® P 1 x 25
13677 Anaerocult® A 1 x 10
14255 Anaeroclip® 1 x 25
1.13675 Anaerotest® 1 x 50
13681 Anaerobic jar 1 ea

Experimental Procedure and Evaluation                      Preparation Quality Control                                                                              
Typical Composition (g/liter) Literature


Mode of Action
 Sulfite Polymyxin Sulfadiazine Agar contains a broad spectrum of nutrients. Sulfite is reduced by most clostridia (including Cl. perfringens) to sulfide, which reacts with iron citrate and causes the colonies to turn black. Other sulfite-reducing microorganisms are largely suppressed by polymyxin and sulfadiazine (sulfapyrimidine). The low sulfite content allows growth of even sulfite-sensitive clostridia which also exhibit an adequate blackening of the colonies (PUT et al. 1961; BEERNS et al. 1961).

Preparation
 Suspend 40 g/liter, autoclave (15 min at 121 °C).
 pH: 7.0 ± 0.2 at 25 °C.
The prepared medium is clear and yellowish-brown.
Experimental Procdure and Evaluation
 Mix the culture medium with the sample material (homogenized and diluted), pour into plates or tubes. Seal the tubes with sterile liquid paraffin. Place the plates in an anaerobic jar. Anaerocult® A, Anaerocult® A mini or Anaerocult® P can be used for this purpose.
 Incubation: 24-48 hours at 35 °C.
Clostridia develop with black colonies. Further tests should be performed for purposes of identification.
Quality control
Test strains
 Growth
 		Black colonies
Clostridium perfringens ATCC 10543
 		good / very good
 		+
Clostridium perfringens ATCC 13124
 		good / very good
 		+
Clostridium sporogenes ATCC 11437
 		good / very good
 		+
Escherichia coli ATCC 25922
 		none / fair
 		-
Pseudomonas aeruginosa ATCC 27853
 		none / poor
 		-

Typical Composition (g/liter)
 Peptone from casein 15.0; yeast extract 10.0; iron(III) citrate 0.5; sodium sulfite 0.5; Polymyxin B sulfate 0.01; sodium sulfadiazine 0.12; agar-agar 13.9.
Literature

ANGELOTTI, R., HALL, H.E., FOTER, M.J., a. LEWIS, K.M.: Quantitation of Clostridium perfringens in Foods. Appl. Microbiol., 10; 193-199 (1962).
 BEERENS, H., CASTEL, M.M., et LECLERC, H.: Contribution ŕ l'étude des Milieux au sulphite de sodium pour l'isoelement des Clostridium. Ann. Inst. Pasteur Lille, 12; 183-193 (1961).
 PUT, H.M.C.: Sulphito-réduction et sulphito-sensibilité des Clostridia: considérations txonomiques et practiques. Ann. Inst. Pasteur Lille, 12; 175-181 (1961).