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The test uses isolated bacterial colonies and is based on in vitro hydrolysis of a carbapenem, imipenem.
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TU were determined by counts of the number of bacterial colonies and were expressed either as TU/µl or as Relative TU.
The plates were scraped to recover the bacterial colonies and were extracted using the phospholipid extraction procedure of Bligh and Dyer [20] as modified by Garbus et al. [21].
Overnight cultures were started from a single bacterial colony and were grown overnight in liquid LB at 37°C with shaking.
For the rapid detection and isolation of PHB-producing bacteria, 0.02% alcoholic solution of Sudan black B was applied to stain bacterial colonies and the plates were kept undisturbed for 30 min.
Fluorescently labeled bacterial colonies and surrounding tissue were microdissected and catapulted into the lid of a 0.2 ml tube containing 25 μl extraction buffer from the Arcturus PicoPure RNA Isolation Kit (Life Technologies).
Total bacterial colonies and count of each species were obtained.
After cloning into pKSU6, random bacterial colonies were picked, and PCR was performed to examine the inserts.
Thus, physiological heterogeneity within a bacterial colony and pharmacokinetic limitations are possible explanations for the frequent failure of antibiotic treatment alone in the management of infectious scleritis [18 20].
The morphology of the resulting bacterial colonies was inspected and confirmed to be S. aureus.
The mucoid appearance of Xap and Xff bacterial colonies is an indication of xanthan production, which is under the regulation of the diffusible factor DSF (our unpublished data).
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