Pseudomonas Agar F, Base

Pseudomonas Agar F Base

EMD Cat. No. 1.10989.0500

500 g

Elective culture media proposed by KING, WARD and RANEY (1954) for the isolation and differentiation of Pseudomonas based on the formation of pyocyanin and/or pyorubin or fluorescein.

These media comply with the recommendations of the United States Pharmacopeia XXIII (1995) and correspond to the culture media specified in the DIN Norm 38411 (examination of water).

Additives

Mode of Action	Preparation
Experimental Procedure and Evaluation	Quality Control
Typical Composition (g/liter)	Literature
Picture/Image

Mode of Action
Pseudomonas Agar P favours the formation of pyocyanin and/or pyorubin and reduces that of fluorescein, whereas Pseudomonas Agar F stimulates the production of fluorescein and reduces that of pyocyanin and/or pyorubin. Simultaneous use of both culture media allows rapid, preliminary identification of most Pseudomonas species, as some strains can only synthesize pyocyanin, some form only fluorescein and others produce both pigments.

Preparation
Suspend 10.0 ml glycerol/liter together with 35 g Pseudomonas Agar F Base/liter or 44 g Pseudomonas Agar P Base/liter, dispense into test tubes if desired, autoclave (15 min at 121 °C). Make slant tubes or pour plates.
pH: 7.2 ± 0.2 at 25 °C.
The plates are clear to opalescent and yellowish-brown (1.10989.); clear and yellowish-brown (1.10988).

Experimental Procedure and Evaluation
Inoculate the surface of the culture medium with cultures suspected to contain Pseudomonas so that individual colonies develop.
Incubation: up to 7 days at 35 °C.

Check for bacterial growth after 24, 48 and 72 hours and then after 6 days.
Pseudomonas aeruginosa can grow on Pseudomonas Agar P to form colonies surrounded by a blue to green zone due to pyocyanin formation or with a red to dark brown zone due to pyorubin production. The colored pigments can be extracted with chloroform. Pseudomonas aeruginosa appears on Pseudomonas Agar as colonies surrounded by a yellow to greenish-yellow zone resulting from fluorescein production. If pyocyanin is also synthesized, a bright green color is produced which fluoresces under UV light.
According to BLAZEVIC et al. (1973), atypical pyocyanin-negative, fluorescein-positive Ps. aeruginosa strains can be differentiated from Ps. fluorescens and Ps. putida. BRODSKY and NIXON (1973) reported that the fluorescence of Ps. aeruginosa colonies in ultra-violet light following growth on MacCONKEY agar can be exploited to provide a rapid orientation test, Ps. fluorescens and Ps. putida do not fluoresce and show only scanty growth.

Quality control of Pseudomonas Agar F

Test strains	Growth	Yellow-green pigment in daylight	Fluorescence at 366 nm
Pseudomonas aeruginosa ATCC 27853	good / very good	+	+
Pseudomonas aeruginosa ATCC 9027	good / very good	+	+
Pseudomonas fluorescens ATCC 17397	good / very good (48 h)	±	±
Aeromonas hydrophila ATCC 7966	good / very good	-	-
Escherichia coli ATCC 25922	good / very good	-	-
Enterobacter cloacae ATCC 13047	good / very good	-	-

Quality control of Pseudomonas Agar P

Test strains	Growth	Blue-green pigment in daylight
Pseudomonas aeruginosa ATCC 27853	good / very good	(+)
Pseudomonas aeruginosa ATCC 9027	good / very good	+
Pseudomonas aeruginosa ATCC 25668	good / very good	+
Pseudomonas fluorescens ATCC 13535	good / very good	- (yellowish)
Aeromonas hydrophila ATCC 7966	good / very good	-
Escherichia coli ATCC 25922	good / very good	-
Enterobacter cloacae ATCC 13047	good / very good	-

Pseudomonas Agar F Base:

Typical Composition (g/liter)

Peptone from casein 10.0; peptone from meat 10.0; magnesium sulfate 1.5; di-potassium hydrogen phosphate 1.5; agar-agar 12.0.

Also to be added:

glycerol 10.0 ml.

Pseudomonas Agar P Base:

Typical Composition (g/liter)

Peptone 20.0; magnesium chloride 1.4; potassium sulfate10.0; agar-agar 12.6.

Also to be added:

glycerol 10.0 ml.

Literature

BLAZEVIC, D.J., KOEPCKE, M.H., a. MATSEN, J.M.: Incidence and identification of Pseudomonas fluorescens and Pseudomonas putida in the clinical laboratory. Appl. Microbiol., 25; 107-110 (1973).
BRODSKY, M.H., a. NIXON, M.C.: Rapid method for detection of Pseudomonas aeruginosa on McCONKEY-Agar under ultraviolet light. Appl. Microbiol., 26; 219-220 (1973).
DIN Deutsches Institut für Normung e.V.: Deutsche Einheitsverfahren zur Wasser-, Abwasser und Schlammuntersuchung. Mikrobiologisches Verfahren (Gruppe K). Nachweis von Pseudomonas aeruginosa (K 8). DIN 38411.
GEORGIA, F.R., a. POE, C.F.: Study of bacterial fluorescence in various media. I. Inorganic substances necessary for bacterial fluorescence. J. Bact., 22; 349 (1931).
GEORGIA, F.R., a. POE, C.F.: Study of bacterial fluorescence in various media. II. The production of fluorescence in media made from peptones. J. Bact., 23; 135 (1932).
KING, E.O., WARD, M.K., a. RANEY, D.E.: Two simple media for the demonstration of pyocyanin and fluorescin. J. Lab. Clin. Med., 44; 401-307 (1954). United States Pharmacopeia XXIII, Chapter "Microbial Limit Tests", 1995.

Pseudomonas Fluorescens ATCC 17397

Pseudomonas Agar F (Base) - Pseudomonas aeruginosa