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(microsoft powerpoint - listeria fraser broth [\310\243\310\257 \270\360\265\345])

Medium for secondary enrichment of Listeria spp.
Tryptone ………………………………………………………………………… 5.0Peptone ………………………………………………………………………….5.0Lithium Chloride ……………………………………………………………….3.0Beef extract ……………………………………………………………………. 5.0Yeast extract ……………………………………………………………………5.0Sodium Chloride …………………………………………………………….20.0Di sodium Hydrogen Phosphate, anhydrous ……………………….9.5Monopotassium Phosphate ……………………………………………… 1.35Esculin …………………………………………………………………………… 1.0Nalidixic Acid …………………………………………………………………. 0.02Acriflavine …………………………………………………………………….0.025Final pH = 7.4± 0.2 at 25°C.
Suspend 47.9 g of powder in 1 litre of distilled or deionized water. Heat until completely dissolved. Sterilize inautoclave at 121°C for 15 minutes. Cool to 45-50°C. Aseptically add 2 vials of Listeria Fraser Supplement(Ferric ammonium citrate 250 mg/vial) (MB-L1166). Mix well. Dispense into sterile final containers.
agents in the medium are acriflavine, an acridinic derivative with bacteriostatic characteristics towards Gram-positive bacteria and nalidixic acid which inhibits Gram-negative bacteria: the concentrations are doublerespect to DEMI FRASER BROTH. The presence of Listeria in LISTERIA FRASER BROTH is indicated by theblackening of the culture caused by a reaction of the esculetin, produced by the aesculin hydrolysis, with theferric ions.
Inoculate 10 ml of Listeria Fraser Broth with 0.1 ml of the primary enrichment broth (i.e. UVM 1 enrichmentbroth) which has been incubated for 20 to 24 hours. Incubate at 36 ± 1°C for 26 ± 2 hours (ensure at least24 hours of incubation). Compare each inoculated tube with an uninoculated control against a whitebackground. Tubes that darken or turn black should be subcultured onto Oxford medium or Palcam medium.
Tubes that retain the original yellow colour should also be inoculated on plating media and confirmed as freefrom Listeria spp before discarding.
Dehydrated mediumAppearance: free-flowing, homogeneous.
Colour: tan.
Prepared mediumAppearance: slightly opalescent.
Colour: light amber.
Incubation conditions: 36 ± 1°C /26 ± 2 hours.
Kisan Biotech Co., Ltd. (OEM Manufactured)2F, Kisan B/D, 86-2, YangJae-Dong, SeoCho-Gu, Seoul #137-135 South KoreaTEL: 82-2-529-2282 FAX: 82-2-529-2284 www.kisanbio.com , www.KSBio.com The powder is very hygroscopic: store the powder at 10-30°C, in a dry environment, in its original containertightly closed and use it before the expiry date on the label or until signs of deterioration or contamination areevident. Store prepared tubes at 2-8°C.
1. Fraser, J.A., and W.H. Sperber (1988) . J. Food Prot , 51, 762-765.
2. Ordinanza ministero della Sanità del 07-12-93: Limiti di Listeria monocytogenes in alcuni prodotti alimentari. Gazzetta Ufficiale della Repubblica Italiana n.291 del 13-12-93.
3. NF V08-055: December 1993, Food microbiology. Detection of Listeria monocytogenes. Routine method.
Kisan Biotech Co., Ltd. (OEM Manufactured)2F, Kisan B/D, 86-2, YangJae-Dong, SeoCho-Gu, Seoul #137-135 South KoreaTEL: 82-2-529-2282 FAX: 82-2-529-2284 www.kisanbio.com , www.KSBio.com

Source: http://www.kisanbio.com/design/kisanbiotech/PDF/LISTERIAFRASERBROTH.pdf

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