I'm working with the enrichment of an organic compost with PGPR, but this compost is not sterilized. How could I quantify viable cells of Azospirillum brasilense, Bacillus subtilis, and Pseudomonas fluorescens in this organic compost 30 days after inoculation? Are there selective media? Is there any other method? For Azospirillum, could I use an indirect method, as NFb?
Cheers,
Artur
Generally you can quantify by serial dilutions by quantitative means. Regarding to viable cells species, you must have to isolate specific media and pick up the prominent viable cells and observe growth by treaking method and it's increasing size. Then prepare pure culture slant without contamination and identify phylogenetic tree construction by using 16S rRNA gene sequencing under PCR for perticular species identification.
This answer can help you..... thanks.
Hi Artur,
I recommend you first perform an antibiogram (antibiotics resistance to your bacterial strains). Find the antibiotics whose your bacterial strains are resistant. and combine them in one selective Medium. Of course, you may have many selective media depending on your results. and then do the plate streaking method to the enriched compost with your strains. This method is not highly accurate but it gives you an idea regarding your amount of your viable bacterial population added to the compost.
I hope this answer brought some help.
Good luck
Generally you can quantify by serial dilutions by quantitative means. Regarding to viable cells species, you must have to isolate specific media and pick up the prominent viable cells and observe growth by treaking method and it's increasing size. Then prepare pure culture slant without contamination and identify phylogenetic tree construction by using 16S rRNA gene sequencing under PCR for perticular species identification.
This answer can help you..... thanks.
Dear Artur,
As the question says, you have to quantify your microbes of interest in unsterilized compost, one of the best ways is molecular tagging of your microbes and then tracking in the unsterilized compost. Since compost is rich in microbial diversity, other traditional cultural approaches may not be that efficient.
Fluorescence imaging is also an approach that could be followed (e.g.FITC-WGA probes; FISH probes).
Alternatively, ARA (antibiotic resistance analysis) could be done to differentiate the microbe of your interest with other microbes, but again, difficult to find a unique antibiotic resistance profile of microbes of your interest.
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