L.K.V. (LAKED SHEEP BLOOD KANAMYCIN VANCOMYCIN) AGAR

Technical Data #1408a / 2013.05.07

LKV Agar is a selective solid medium used for cultivation and isolation of Bacteroides sp. Laked blood permit to identify Bacteroides by their specific fluorescence activity. Antibiotics inhibit gram-positive and some anaerobic gram-negative bacteria. Hemin and vitamin K are growth factors for Bacteroides.

 


 

FORMULA
in grams per liter purified filtered water

Casein Peptone

10,0

Meat Peptone

5,0

Yeast Extract

5,0

Heart Peptone

3,0

Sodium Chloride

5,0

Starch

1,0

Agar

13,5

Enrichments

 

Hemin

0,005

Vitamin K

0,01

Laked Sheep Blood

50 ml

Antibiotics

 

Kanamycin

0,1

Vancomycin

0,0075

pH 7.3 +/- 0,2 25 C

This approximate formula may be adjusted and/or enriched to obtain best results.


PRECAUTIONS
This medium is for IN VITRO diagnostic use only.


STORAGE
Store prepared media at 2-8 C protected from direct light and dehydrated powder, in a dry place, in tightly-sealed containers at 2-25 C.


SIGN OF DETERIORATION
Media should not be used if the expiry date has passed. Prepared media should not be used if there are signs of contamination or deterioration (shrinking, cracking, evaporation or discoloration). Do not use dehydrated media if it is caked.


DIRECTIONS
Suspend 42,5 g of dehydrated media in 950 ml of purified filtered water. Heat with frequent agitation and boil for one minute. Sterilize at 121 C for 15 minutes. Cool to 45-50 C and aseptically, add 50 ml of Laked Sheep Blood (4379), 2x5 ml of Enrichment Vitamin K-Hemin (8752) and 2x5 ml of antibiotics Kanamycin-Vancomycin 2x5 ml(8732). Mix gently and dispense into sterile Petri dishes.


PROCEDURE
Prior to inoculate, the prepared media should be brought to room temperature.

  1. Suitable anaerobes recovery is depending of collection and transportation of specimens. Specimens should be transferred into Anaerobes Pre-Reduced Transport Medium (2037) immediately after collection and brought to the laboratory quickly.
  2. Directly, inoculate plate as soon as possible with a large amount of inoculum, streak for isolation.
  3. Incubate at 35 C under anaerobic conditions for 48 hours.
  4. Examine for growth and fluorescence.


QUALITY CONTROL

Results after 48 hrs at 35C anaerobic atmosphere

Organisms

ATCC

Growth

Bacteroides fragilis

25285

+

Bacteroides thetaiomicron

29741

+

Clostridium perfringens

13124

-

Staphylococcus aureus

25923

-

Escherichia coli

25922

-


LIMITATIONS OF METHOD
This medium is only a part of the identification. Other tests may be required.


REFERENCES

  1. Dowell, V.R.Jr., G.l. Lombard, F.S. Thompson, A.Y. Armfield 1977 Media for isolation, characterization and identification of obligately anaerobic bacteria. CDC Laboratory Manual. Center for Disease Control, Atlanta
  2. Lennette, E.H., Ballows, A., Hausler, W.J.Jr., and Shadomy, H.J. Manual of Clinical Microbiology. 4th ed. 1985 Washington D.C.: American Society for Microbiology.
  3. Sutter, V.L., D.M. Citron, M.A.C. Edelstein, S.M. Finegold Wadsworth Anaerobic Bacteriology Manual 4th ed C.V. Mosby St. Louis
  4. Van Winkelhoff, A.J., J. De Graaf 1983 Vancomycin as a selective agent for isolation of Bacteroides. J. Clin. Microbiol. 18:1282-1284
  5. Wilkens, T.D., S.L. Chalgren, F. Jimenez-Ulate, C.R. Drake Jr, J.L. Johnson 1976 Inhibition of Bacteroides fragilis on blood agar plates and reversal of inhibition by added hemin J. Clin. Microbiol. 3:359-363


CATALOGUE NUMBERS

Dehydrated media 500 g QB-39-1006
     
Prepared media    
Plates, 100x15 mm 10/pkg 1408
Mono-Plates, 100x15 mm 10/pkg 1408M
     
Enrichment Vitamin K-Hemin, 5ml 10/pkg 8751
     
Antibiotics    
Kanamycin-Vancomycin, 5ml 10/pkg 8732
     
Laked Sheep Blood 50 ml 4379

 


 

 

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