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Counting Bacteria.

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Presentation on theme: "Counting Bacteria."— Presentation transcript:

1 Counting Bacteria

2 Serial Dilutions Because of their very ______ ______, counting the number of bacteria in a sample can be _________ Although _______ _______ are possible with a ______________, they require a lot of time and ___________

3 Why Count the Number of Bacteria??
________ _____________: How effective drugs are in killing microorganisms? ___________________: How many MO in CSF, blood, urine ___________ ____________: - Contaminants in food, milk, water

4 Total Cell Count The most common method of _______________ the total microbial cells (______ ___ _________) The ______ ___________ of cell suspension in a __________ _____________ of known _______ using a ________________

5 Total Cell Count – Both living and dead MO
Disadvantage: ____________________________________________________________________________________________________________ Advantage: _____________________________________________________________________________________________________________

6 Total Cell Count – Both living and dead MO
Neubauer ___________ ___________ Special _______ ____ _______ _______, microscopic marks on slide, known _________

7 Counting Chamber

8 Counting Chamber Use special ______________ ____ _____________________
Count specific areas of __________________, multiply by __________________ ______________

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10 Instrumentation * _____________________ (Light Meter)
Each single ____ stops some ______ from ____________________________________________ One side _____ ________, then test tube containing MO, other side _______ ________________ Needle reading from scale, then ________________ _____________ telling # of __________________________________________

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14 Coulter Counter Another method for _____________ _____________
An electronic instrument “_____________ _________” Has a tiny tube (_____ ______ _____________) that lets through only one cell at a time

15 Viable Cell Count – Living Cells Only

16 Viable Count – Living Cells only
An easier method is to ______ ________ _____ __ _____ ______ (nutrient agar plate) and count the number of ________ _____ ______ If the bacteria are ______ ____ ______, each bacterial cell in the original sample should produce a _______ _______

17 Serial Dilutions Bacterial samples must be _______ considerably to obtain ___________ ___________ To determine the ________ ___ ______ in a bacterial culture carry out _______ ____________

18 Serial Dilutions Bacterial cell numbers are usually _______ _____ _____________________________ Plating out this sample, undiluted, would lead to the creation of a ___________________ Bacterial Lawn - many individual bacteria colonies that are ______________________ ____________________________________

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20 Serial Dilutions Bacterial cell numbers _____ __ __ _______
This is done by ______ ________ the amount of bacteria you have in your sample A ______ _________ of bacteria sample is mixed with a _____ _______ (sterile broth or water), and then _______ ___________ are made

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22 Serial Dilutions A small amount of _____ ___ ____ _______ _________ ________ is then spread onto an ______ ________ The numbers of bacterial colonies that ______________________________

23 Serial Dilutions Work ___________ using multiplication with the “___________ ____________” ________ ________ - the ______ ___ ______ that you have diluted the bacteria sample with the diluent solution Make a determination of the numbers of bacteria in ______ ________ _________.

24 Serial Dilution

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26 Serial Dilutions *Make ________ _____________
*Spread a small amount from each tube onto _______ _______ *Culture bacteria for a ________ __ _________ *Count colonies ________________________________

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29 Membrane Filtration ________ __________ can also be used to determine the ________ _______________ In this method ______ _____ ___________ onto membrane filter which is then placed over a ________ ______ __________.

30 Membrane Filtration The methodology is similar to conventional _______ ________ _______________. Membranes have a printed millimeter ________ and colonies can be counted under a _________ _________________.

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33 Factors essential for __________ ________
--proper ________________ --proper ___________________ (space, temperature) --No inhibiting factors Factors that _____________________ __________________________________________ --lack of _____________ --lack of __________ __________________ --presence of an ___________ __________


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