2.2. Bacterial culture and preliminary identification
The specimens were processed in the laboratory within 2 h for cultivation of aerobic and anaerobic bacteria. Briefly, transport medium containing the fixture was vortexed strongly to release attached cells from the surface of the implant fixture. After vortexing serial dilutions were made and aliquots of 10 μl were inoculated on several non-selective and selective agar media, including cadmium-fluoride-acriflavin-tellurite (CFAT) agar selective for Actinomyces species [24]. The plates were incubated for aerobic bacteria in an atmosphere containing 5% CO2 for 2–3 d and for anaerobic bacteria in anaerobic jars filled with a gas mixture (80% N2, 10% H2, 10% CO2) for 5–7 d. The total viable aerobic and anaerobic growth was determined as the total number of colony forming units (CFU) on non-selective blood and Brucella blood agar plates, respectively. Isolates representing different colony morphologies, detected from both non-selective media and CFAT-agar, were presumptively identified to bacterial genus/group level by established biochemical methods [25] and stored at −70 °C in vials containing 20% sterilized skim milk until further testing.
2.2. Bacterial culture and preliminary identification
The specimens were processed in the laboratory within 2 h for cultivation of aerobic and anaerobic bacteria. Briefly, transport medium containing the fixture was vortexed strongly to release attached cells from the surface of the implant fixture. After vortexing serial dilutions were made and aliquots of 10 μl were inoculated on several non-selective and selective agar media, including cadmium-fluoride-acriflavin-tellurite (CFAT) agar selective for Actinomyces species [24]. The plates were incubated for aerobic bacteria in an atmosphere containing 5% CO2 for 2–3 d and for anaerobic bacteria in anaerobic jars filled with a gas mixture (80% N2, 10% H2, 10% CO2) for 5–7 d. The total viable aerobic and anaerobic growth was determined as the total number of colony forming units (CFU) on non-selective blood and Brucella blood agar plates, respectively. Isolates representing different colony morphologies, detected from both non-selective media and CFAT-agar, were presumptively identified to bacterial genus/group level by established biochemical methods [25] and stored at −70 °C in vials containing 20% sterilized skim milk until further testing.
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