-How to culture it on MRS media?
- i am student and due to costs, instead of buying MRS broth and agar, I want buy MRS broth and prepare MRS agar from it, is it possible?
-what is the culturing condition for Lactobacilli?
- further I want work on lactobacillus L. crispatus, L. gasseri, and L. jensenii and identification by molecular technique, is it possible?
Hello, you can use swab and grow in MRS broth tube at 30*C, till you will observe turbidity.
After you can transfer the 0.5-1ml to 90mm plate with MRS agar. After grow culture petter to perform microscopy to differentiate the lactococcus from lactobacilli. After you can perform API test for identification of cultures.
Solhan Mustafa
1. Yes. Possible. You can add 1.2% agar with MRS broth to make MRS agar.
2. You may incubate at 37 degree Celsius.
3. Definitely, you can identify by using 16s rRNA sequencing.
Of course you can. however i think it better, if you isolate onto de Man, Rogosa and Sharpe (MRS) supplemented with 0.05% L-cysteine. Article Isolation of Vaginal Lactobacilli and Characterization of An...
Yes, it's absolutely possible to prepare MRS agar from MRS broth. MRS broth is a specialized medium designed to support the growth of Lactobacillus species. Add a solidifying agent like agar to transform it into a solid medium (MRS agar) that's essential for isolating and culturing these bacteria.
Lactobacilli are generally microaerophilic or anaerobic bacteria, meaning they prefer environments with low oxygen levels. Typical culturing conditions:
- 37°C (98.6°F)
- Anaerobic or microaerophilic conditions (can be achieved using anaerobic jars or gas packets)
- pH: 5.5-6.5 (slightly acidic)
Identifying Lactobacillus species using molecular techniques is a common and reliable method (not cheap):
Common molecular techniques for Lactobacillus identification: 16S rRNA sequencing, Species-specific PCR, Multilocus sequence typing (MLST), which are not cheap. Sorry but if you are a student and budget limited, you won't be able to go through it.
Some more affordable alternatives are Phenotypic Methods including:
- Biochemical Tests: They assess the ability of bacteria to utilize or produce specific substances. While they may not be as specific as molecular methods, they can provide valuable information about the species. Examples include catalase, oxidase, and carbohydrate fermentation tests.
- Gram Staining: Lactobacillus species are Gram-positive.
Also, depending on the region you live at you can find molecular methods with Lower Costs like:
- PCR with Species-Specific Primers: Designing and synthesizing primers specific to certain Lactobacillus species can be less expensive than whole-genome sequencing.
- Rapid Diagnostic Kits: Commercially available kits often offer faster turnaround times and may be more cost-effective than traditional laboratory methods.
While these methods may be cheaper, they might not always be as accurate as molecular sequencing. Phenotypic methods can be time-consuming, especially if multiple tests are required and some methods may require specific equipment or reagents that can add to the overall cost.
Hope it helps!