Detection of Escherichia coli (E. coli)_A Complete Procedure (ISO 9308-1 & ISO 16649)
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- Опубліковано 26 чер 2024
- Detection of Escherichia coli is a very important Microbiological testing parameter for Food, Feed, Water and Environmental samples analysis. This video represents a detailed procedure of for the detection of E. coli in a sample.
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LyRical WoRLD Sachin Yadav, you are most welcome. Thanks for being with us.
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@@MicroChemsExperiments I wish I could have known your channel when I was in MTLA Program 😁 I come from Indonesia but been 2 years in Germany. How about you? I was wondering if you can also make a video about "Plaque reduction neutralization test" and more bacteriology test including the biochemical test. It would be really helpful for us who cant bring the notes to another country 😊
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Please, what sample was used?
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Hello dear. I have a question in cca agar plate how we can differentiate ecoli and coliform because cca is suitable for both e coli and coliform
Is this a Most Probable Number (MPN) method or Membrane Filtration Method (MFM)? Can it work for turbid water?
I have a one question from you...
I collected sample from the sewage water,and followed with 3 times serial dilution with bglb broth and incubated for the identification of coliform bacteria.after the production of gas I isolate them into(0.1ml) macconkey agar and did indole test for them for the confirmation.in plates I've got 60 colonies,so the CFU /ml was calculated...6x10^5 CFU /ml ...so,I need to dilute them into three samples.like 10 CFU/ml,100 CFU/ml and 1000 CFU/ml...so can u give me an idea how to do that....
And my second question after diluted them,I need to those cultures to grow inside the nutrient broth for the DNA extraction,so,how can I inoculate them,I mean after getting dilution,we should do the culture again which I mention in 1st question like culture into the agar plates,if I take the one colony from the agar plate define 1 ml?I mean let say we culture the agar plate from the 100cfu/ml sample if I take one colony from them means that colony contain 100cfu?
Eelam Music Production, Its soo tough to explain here to solve your question. But I'm trying....
Situation-1: You got 60 CFU from 0.1 BGLB broth culture
****Ans to the first question:
1. To get 10CFU/ml sample, dilute 1ml BGLB culture (600CFU) into 59ml normal saline.
2. To get 100CFU/ml sample, dilute 1ml BGLB culture (600CFU) into 5ml normal saline.
3. To get 1000CFU/ml sample, you need to use lower dilution in the same way as done for 1 & 2.
****Ans to the second queation:
A single colony contains millions of CFU. First dilute 1 colony in 100ml normal saline then culture 0.1ml from the diluted colony on nutrient agar. Count the CFU on nutrient agar plate.
Now Calculate and dilute the normal saline as much as you need to find your desired bacterial concentration, on the basis of how much CFU you got on NA plate.
If you dont get the expected result by following my guideline then try the bellow procedure:
First culture a coliform bacteria in nitrient broth, centrifuge at 4000rpm for 5 min. Discard the upper broth layer, dilute the lower bacterial cell in normal saline, prepare McFarland 1.5 turbidity and match the turbidity of your bacterial cell using a spectrophotometer. Then dilute to desired concentration of bacterial cell as needed.
Thank you.
sorry can you give the specifications of EMB AGAR MEDIA you used?
if we have a swab sample, how to prepare a sample?
after we strake , it fills the petri dish, not in scratches, is that contamination? Or too much bacteria?
In this, you had prepared peptone salt solution, did you not use it in this entire procedure? Can you tell why was it prepared in this procedure?🤔
Hi, thank you for the video.
May I ask for the peptone salt solution, can it be replaced with Buffered peptone water (BPW)? Also, I would like to ask is there a difference if I use 0.1% BPW, instead of BPW?
You can use BPW as diluent. No problem with it
@@MicroChemsExperiments thank you
Thank you for the video. Is there a method for E coli enumeration?
Rbar H, You are most welcome. We will prepare video for the
enumeration of E. coli soon. You can enumerate E. coli by following steps:
1. Dilute sample to appropriate dilution.
2. Spread 0.1ml diluted sample on CCA plate.
3. Consider purple colonies as presumptive E. coli.
4. Confirm E. coli colonies by growing on EMB and sMac plates.
5. Count confirmed colonies as CFU.
To get the video, please wait. We will work on it soon.
Rbar H, please find the E. coli enumeration video. Video Link: ua-cam.com/video/aBEoNeboAHs/v-deo.html
I have a doubt , for inoculation of E. coli on agar , we should use only one loopful of ecoli broth or for each quadrant we use individual loopful of ecoli
You should use one loop full of E. Coli and draw the first quadrant. That's enough. No need to take again for other quadrant
Could you tell what color colonies to check for salmonella, staphylococcus, pseudomonas with their respective agar medium. Also rest of the procedure would remain same ?
We will make video soon
In the procedure how to calculate the e coli?
For CCA, it don't need to be autoclaved before it is pour into the petri dish?
Siti Salihah Mohd Saidi, No. Because CCA is highly selective for E. coli and Coliform and high temperature (121 degree) may destroy the selective ingredients of the medium. That's why, no need to autoclave the medium during the media preparation. Just boil to dissolve the content and pour into petri dish.
@@MicroChemsExperiments aaaa I see, thank you for the explanation
@@sitisalihahmohdsaidi2583 You are most welcome. Thanks for being with us.
Hello. I have a question. If we use this microbiological test for E.coli detection, do we also need to perform biochemical test?
Only perform Oxidase Test along with gram staining if you have facilities. Other biochemical test is not mandatory according to ISO reference method which is mentioned in this video.
@@MicroChemsExperiments Noted. Thank you so much !
Indole test can be performed?
❤️
Thanks
Can I ask one question is this possible that your place is not cleab but still your coliform count is absent .
Yes. This is possible.
@@MicroChemsExperiments how it is possible I am microbiologist I have done testing in certain sample where I have found colliform but the same sample when tested outside they didn't detect any coliform...can you plz sort out my quiery
Considering EMB and sMAC is used after 48 hours, is there need to prepare it on first day?
Not necessarily
if sorbitol agar we dont have so we use mackonkey agar ?
Yes. Use MacConkey Agar
I am a research student so can you please share the articles because I follow this video for the detection of E.coli. Please also explain what kind of the feed meal you use as a sample.
We have followed ISO 9308 & ISO 16649 to make this video. Please collect the handbook from ISO as we do not provide any pdf.
In this video, we have tested a poultry feed meal sample.
@@MicroChemsExperiments Thank you so much.
@@entertainmentpoint4242 It's pleasure
Can we use this method for E.coli detection from hand?
You can use
Can we use this method for E.coli detection from water sample..
Yes. You can
Dear sir I want all microbiological tests videos for drinking water
Okay. Stay with us
What about double strength mac conky agar
We will make video later
E coli O157 Enrichment Medium
Can buffered Peptone Wtaer with Pyruvate (bPWp) and ACV (Acriflavin-Cefsoludin-Vancomycin) be used ?????
Just use Selective media for E coli O157.
My Qus.
pri-enrichment medium name ??
and Enrichment medium name??
which pri Enrichment and Enrichment medium use for detection of E coli o 157 H:7 test .??
@@ajitpatil2153 Enrichment Broth: mTSB with Novobiocin.
Pre-enrichment is not necessary
Thanks
@@ajitpatil2153You are welcome
thanks
can i get pdf for this test,?
Bokul Hossain, we combined two ISO methods with few modifications for E.coli test to simplify and strongly confirm the E.coli cell. You can't match full method to any of the refetence methods.
Please give calculation part
Wich sample you give for the this test...plz tell me
Prajwal Nimbulkar, we tested a feed meal sample in this video
@@MicroChemsExperiments thanks for replying
Thankyou so much
@@prajwalnimbulkar267 Its pleasure. Stay with us.
Name of this type of method ????
The reference of the method: ISO 9308-1
Biochemical tests
No need as per ISO SOP.
Where are u from mam ??
We should not discuss it. Please ask questions related to the video
If you have maual for thia i want
Sorry. We don't provide any document
How to detect second strain e coli O157 H:7 ?????
plzz reply
Firstly, analyze you sample according to this video. If you get the E.coli, streak several bacterial colony on "HiCrome EC O157:H7 Selective Agar Base" selective agar plate. Finally if you find dark purple to magenta coloured colony then you can interpret your test result of e coli O157 H:7 as detected.
@@MicroChemsExperiments e coli o157 h:7 selective media name plzz ??
@@ajitpatil2153 Already mentioned in the first reply. The name is: HiCrome EC O157:H7 Selective Agar Base
Did u do this practical for o157 Ecoli
How much tissue need for the purpose of study?😢
Can we do internship here
No sir.
How to prepare the result and report to the given sample
Aanchal Gahalot, the result is E. coli detected.
The the result should be reported as 'Detected/25g'
Please make video without music.
Okay
Is this biochemical test ?
No sir. This is Microbiological test
@@MicroChemsExperiments you have any video or PPT on identification of E.coli by biochemical testing .
Because I am University student
And I need this PPT for presentation
@@hassaan7990 No PPT is available. Search on Google
@@MicroChemsExperiments actually I searched it . But I could not find it. Can you please help me 🥺 for collecting the data .
@@hassaan7990 No sir. we can't help at this moment. Sorry
thankful for all video it is helpful am very interesting for more help
Thank you
Hi
Hello
Please give me the following information
detection of E- coli o157 H:7
Pri- enrichment medium name -??
Enrichment medium name -??
Already mentioned