The tubes containing the pulp samples were shaken in a tube shaker for 30 s to disperse bacterial aggregates.
Next, 100 μl of bacterial sample was spread, in duplicate, on the following solid media: trypticase soy agar (HiMedia Laboratories, Mumbai, Maharashtra, India) supplemented with 5% sheep blood for total viable microorganism count, Rogosa Agar (HiMedia Laboratories, Mumbai, Maharashtra, India) for Lactobacillus sp. Count, Schaedler K-V Agar (HiMedia Laboratories, Mumbai, Maharashtra, India) for Prevotella sp. count and mitis salivarius agar (MSA) (HiMedia Laboratories, Mumbai, Maharashtra, India) for Streptococcus sp. count.[18 (link)]
The plates were incubated under anaerobic conditions for 1 week, whereas the MSA plates were incubated in an atmosphere of 5% CO2 for 48 h.
After incubation, microbial counts were performed with a digital colony counter (HiMedia-LA660). Cell morphology was evaluated by Gram staining technique.
Next, 100 μl of bacterial sample was spread, in duplicate, on the following solid media: trypticase soy agar (HiMedia Laboratories, Mumbai, Maharashtra, India) supplemented with 5% sheep blood for total viable microorganism count, Rogosa Agar (HiMedia Laboratories, Mumbai, Maharashtra, India) for Lactobacillus sp. Count, Schaedler K-V Agar (HiMedia Laboratories, Mumbai, Maharashtra, India) for Prevotella sp. count and mitis salivarius agar (MSA) (HiMedia Laboratories, Mumbai, Maharashtra, India) for Streptococcus sp. count.[18 (link)]
The plates were incubated under anaerobic conditions for 1 week, whereas the MSA plates were incubated in an atmosphere of 5% CO2 for 48 h.
After incubation, microbial counts were performed with a digital colony counter (HiMedia-LA660). Cell morphology was evaluated by Gram staining technique.
