BPL Agar (Brilliant-green Phenol-red Lactose Agar acc. to KAUFFMANN)

Brilliant Green Phenol-Red Lactose Agar (BPL Agar)

EMD Cat. No. 1.07236.0500

500 g

Selective agar proposed by KAUFFMANN (1935) for the identification and isolation of Salmonella with the exception of S. typhosa in faeces, urine, meat, milk and other materials.

This culture medium complies with the Fleischbeschaugesetz (German Meat Inspection Law) and the Einfuhruntersuchungs-Verordnung (German Regulations for the Examination of Imported Goods).

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

Mode of Action
This culture medium contains lactose, whose degradation to acid is indicated by the pH indicator phenol red, which changes its color to yellow. The indicator exhibits a deep red color in the alkaline range. The growth of the accompanying Gram-positive microbial flora, Salmonella typhi and Shigella is largely inhibited by brilliant green. ADAM (1966) recommended that 0.2 % sodium deoxychlolate should be added to the culture medium to inhibit the swarming of Proteus colonies.

Typical Composition (g/liter)
Peptone from meat 7.0; sodium chloride 5.0; lactose 15.0; phenol red 0.04; brilliant green 0.005; agar-agar 13.0.

Preparation
Suspend 40 g in 1 liter of purified water. Autoclave (15 min at 121°C), pour plates.
pH: 6.5 ± 0.2 at 25°C.
The plates are clear and red-brown/greenish.

Experimental Procedure and Evaluation
Inoculate the surface of the culture medium massively using either the sample material itself or material taken from an enrichment culture.
Incubation: 18-24 hours at 35°C aerobically.

Tests with less inhibitory culture media such as SS Agar, LEIFSON Agar, ENDO Agar or GASSNER Agar should also be carried out.

Appearance of Colonies	Microorganisms
Pale pink, translucent, surrounded by a red zone	Lactose-negative: Salmonella, occasionally Proteus and Citrobacter
Yellow-green, opaque, surrounded by a yellow-green zone	Lactose-positive: If growth with a good E. coli, Enterobacter, Klebsiella. All others are largely inhibited.

Quality control (spiral plating method)

Test strains	Inoculum (cfu/ml)	Recovery rate (%)	Colony color	Culture medium
Salmonella typhimurium ATCC 14028	10³-10⁵	>= 40	pink	red
Salmonella choleraesius ATCC 13312	10³-10⁵	>= 40	pink	red
Salmonella enteritidis NCTC 5188	10³-10⁵	>= 40	pink	red
Escherichia coli ATCC 25922	10³-10⁵	>= 40	yellow-green	yellow
Proteus vulgaris ATCC 13315	10³-10⁵	Not limited!	pink	green
Staphylococcus aureus ATCC 25923	> 10⁵	<= 0.01
Enterococcus faecalis ATCC 33186	> 10⁵	<= 0.01
Bacillus subtilis ATCC 6633	> 10⁵	<= 0.01

Additives

EMD Cat. No.	Product	Pack Size
	GASSNER Agar	500 g
1.02896.0500	LEIFSON Agar (Deoxycholate Citrate Agar acc. to LEIFSON, modified)	5 kg
1.04044.0500	ENDO Agar	500 g
SX0480-2	Sodium deoxycholate	100 g
1.07667.0500	Salmonella-Shigella Agar	500 g

Literature

ADAM, D.: Zusatz von Natriumdesoxycholat zum Brilliantgrün-Phenolrot-Agar nach Kristensen-Kauffmann zur Hemmung des Schwärmvermögens von Proteuskeimen. - Ärztl. Lab. 12; 245-246 (1966).
Deutsches Fleischbeschaugesetz: Ausführungsbestimmungen über die Untersuchung von gesundheitspolizeilicher Behandlung der Schlachttiere und des Fleisches bei Schlachtungen im Inland. Anlage 1 zu § 20 Abs. 4: Vorschriften über die bakteriologische Fleischuntersuchung.
Verordnung über die Untersuchung des in das Zollgebiet eingehenden Fleisches (Einfuhruntersuchungs-Verordnung). Anlage 1 zu § 20 Abs. 1: Untersuchungsverfahren.
KAUFFMANN, F.: Weitere Erfahrungen mit dem kombinierten Anreichungerungsverfahren für Salmonellabacillen. - Z. Hyg. Infekt. Kr., 177; 26-32 (1935).