分离无乳链球菌(B群)
A selective medium for the rapid isolation of Streptococcus agalactiae and other streptococci involved in bovine mastitis.
Typical Formula* | gm/litre |
`Lab-Lemco’ powder | 10.0 |
Peptone | 10.0 |
Aesculin | 1.0 |
Sodium chloride | 5.0 |
Crystal violet | 0.0013 |
Thallous sulphate | 0.33 |
Agar | 15.0 |
pH 7.4 ± 0.2 | |
* Adjusted as required to meet performance standards
POISON - Contains Thallium Salt
Directions
Suspend 41g in 1 litre of distilled water. Bring to the boil to dissolve completely. Sterilise by autoclaving at 115°C for 20 minutes. Cool to 50°C, add 5-7% of sterile bovine or sheep blood, mix well and pour plates.
Description
A selective medium for the rapid isolation of Streptococcus agalactiae and other streptococci involved in bovine mastitis.
Crystal violet or gentian violet and thallium salts have long been used in selective media for streptococci. Haxthausen1 employed a selective crystal violet medium for the isolation of skin streptococci. Bryan2 using gentian violet blood agar, found that the growth of saprophytic milk bacteria was prevented whilst that of streptococci was unaffected. Edwards3 employed a crystal violet aesculin blood agar for the cultural diagnosis of bovine mastitis, whilst McKenzie4 used a medium containing thallium acetate for the same purpose.
Hauge et al.5 described a composite medium containing all the components of modified Edwards Medium.
Aesculin differentiates the negative Streptococcus agalactiae (blue colonies) from aesculin-positive Group D streptococci (black colonies).
Technique
Inoculate the surface of the medium with centrifuged deposits from milk samples and incubate at 35°C.
Look for pale blue colonies which should then be subcultured for further identification tests.
Storage conditions and Shelf life
Store the dehydrated medium at 10-30°C. and use before the expiry date on the label.
Store the prepared plates at 2-8°C.
Appearance
Dehydrated medium: Straw coloured, free-flowing powder
Prepared medium: Straw coloured gel with blue hue