I'm going to work on anaerobic bacteria for the first time. For this organism, THB agar is used. But I have less knowledge on the protocol for growing such bacteria using a CO2 incubator.
Can someone describe the steps to grow this organism with the required conditions and materials?
Thanking you
It is easy, but the most Important thing is the CO2 Incubator with uninterrupted power.
I've worked with S.aureus so I'm not 100% sure how useful I'll be to you but assuming it is similar - you can inoculate and grow your bacteria on blood agar (as it provides enrichment for the growth of the pathogen) using the inoculation loop to make streak plates and then cultivate the Petri dish over night (12-18h) in the incubator at 36-37C degrees. After that time has passed you'll probably already have nice separate colonies to use for your experiments. If you don't - you can prolong that time with another hour or two. Unfortunately, i cannot tell you the exact protocol because in the lab i did my experiments we didn't use the blood broth but LB medium (for treatment and spectrophotometric measurements) or LB with agar (for cultivating, as well as quality control).
Ps: I also found that PYG medium, modified is also recommended for your strain. Here's the protocol for it: https://books.google.bg/books?id=aWsF-e8tPzIC&pg=PA1451&lpg=PA1451&dq=Streptococcus+intermedius+in+petri&source=bl&ots=9IyHbgP1Ud&sig=ACfU3U0mashBYtY8R95Pu_IqLiKysgrP_w&hl=en&sa=X&ved=2ahUKEwisl7P49vXuAhVQ-qQKHUTiDR0Q6AEwEnoECBMQAw#v=onepage&q=Streptococcus%20intermedius%20in%20petri&f=false pages: 1450-1451
S intermedius is oxygen-tolerant but will grow better in an anaerobic atmosphere. A CO2 incubator is not anaerobic (but S intermedius will grow in air or air + CO2). It depends on what investigations you are planning: there are important gene upregs and downregs, and differences in virulence factors, between aerobic and anaerobic conditions for S intermedius.
It can be cultivated on Colombia agar enriched with 5%defibriated horse blood. Avoid using nickel chrome bacteriological loops better use plastic or platinum loops. The media used should be freshly prepared to minimize O2 diffusion. Better incubated Anaerobic. Best of luck
Hi,
I used to grow S. intermedius on fastidious anaerobe agar (FAA) (Lab M™ International Diagnostic Group pic, Bury, UK) supplemented with 5 % (v/v) defibrinated horse blood (TCS Bioscience Ltd., Buckingham, UK). I used Brain-heart infusion (BHI) broth (Oxoid Ltd., Basingstoke, UK) to prepare bacterial suspensions and incubated in a CO2 cabinet (5 % v/v CO2, 20 % v/v H2 , 70 % v/v N2) at 37 °C.
Hope this will help
