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Chapter 5  Microbial Nutrition and Culture cont’d Siti Sarah Jumali (ext 2123) Room 3/14 [email_address]
[object Object],[object Object]
The Requirements for Growth ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
1) pH ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Physical  Factors Required for Bacterial Growth
2) Temperature ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
 
Food Preservation Temperatures
3) Oxygen ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Patterns of Oxygen Use
4) Hydrostatic Pressure ,[object Object],[object Object],[object Object]
5) Osmotic Pressure ,[object Object],[object Object],[object Object]
Plasmolysis
Halophiles ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Responses to Salt
The Great Salt Lake in Utah
Chemical Requirement: Nutritional Factors ,[object Object],[object Object],[object Object],[object Object],[object Object]
[object Object],[object Object],[object Object],[object Object],Chemical Requirements
Chemical Requirements ,[object Object],[object Object],[object Object],[object Object],[object Object]
Chemical Requirements ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Chemical Requirements ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Organic Growth Factors ,[object Object],[object Object]
Preparation of Culture Media ,[object Object],[object Object],[object Object],[object Object]
Agar ,[object Object],[object Object],[object Object],[object Object],[object Object]
Types of Culture Media ,[object Object],[object Object],[object Object]
Types of Culture Media Type of Media  Purpose Chemically Defined  Growth of chemoheterotrophs and photoautotrophs: microbiological assays Complex Growth of most chemoheterotrophic organisms Reducing Growth of obligate anO2 Selective Suppresion of unwanted microbes; encouraging desired microbes Differential Differentiation of colonies of desired microbes from others Enrichment Similar to selective media but designed to increase numbers of desired microbes to detectable levels.
Culture Media ,[object Object],[object Object],[object Object],[object Object]
 
 
Selective, Differential, and Enrichment Media ,[object Object],[object Object],[object Object],[object Object]
Selective Media ,[object Object],[object Object],[object Object],[object Object],Differential Media
Three species of  Candida  can be differentiated in mixed culture when grown on CHROMagar  Candida  plates
Identification of urinary tract pathogens with differential media (CHROMagar)
Enrichment Media ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Culturing Bacteria ,[object Object],[object Object],[object Object],[object Object],[object Object]
Obtaining Pure Cultures ,[object Object],[object Object],[object Object],[object Object]
The Streak Plate Method uses agar plates to prepare pure cultures
The Streak Plate Method Figure 6.11
Anaerobic Culture Methods ,[object Object],[object Object],[object Object]
Anaerobic Jar Figure 6.6
To culture obligate anaerobes, all molecular oxygen must be removed and kept out of medium.  Agar plates are incubated in sealed jars containing chemical substances that remove oxygen and generate carbon dioxide or water
Anaerobic Transfer
An Anaerobic Chamber Figure 6.7
Capnophiles ,[object Object],[object Object],[object Object]
Preserved Cultures ,[object Object],[object Object],[object Object],[object Object],[object Object]
CHAPTER 6: MICROBIAL GROWTH
Growth and Cell Division ,[object Object],[object Object],[object Object],[object Object]
Binary Fission
Binary Fission
Thin section of the bacterium  Staphylococcus , undergoing binary fission
Budding in Yeast
Phases of Growth ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
 
The Lag Phase ,[object Object],[object Object],[object Object],[object Object]
The Log Phase ,[object Object],[object Object],[object Object],[object Object]
Synchronous growth :  A hypothetical situation in which the number of cells in a culture would increase in a stair-step pattern, dividing together at the same rate Nonsynchronous growth :  A natural situation in which an actual culture has cell dividing at one rate and other cells dividing at a slightly slower rate
[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Stationary Phase
Microbes growing continuously in a chemostat
Measuring Microbial Growth ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Direct Methods ,[object Object],[object Object],[object Object],[object Object],[object Object],Serial Dilution
Plate Counts Figure 6.17
Plate Counts ,[object Object],Figure 6.16
2) Counting Bacteria by Membrane Filtration
3) The Most Probable Number (MPN) Method ,[object Object],[object Object],[object Object],[object Object]
4) Direct Microscopic Counts ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
The Petroff-Hausser Counting Chamber
Direct Microscopic Count
Counting colonies using a bacterial colony counter
Bacterial colonies viewed through the magnifying glass against a colony-counting grid
Which of these plates would be the correct one to count?  Why? Countable number of colonies  (30 to 300 per plate )
Indirect Methods ,[object Object],[object Object],[object Object],A Spectrophotometer:  This instrument can be used to measure bacterial growth by measuring the amount of light that passes through a suspension of cells
Turbidity
Turbidity The less light transmitted, the more bacteria in sample.
[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Biosafety Levels ,[object Object],[object Object],[object Object],[object Object],[object Object]
Biosafety Level 4 (BSL-4) Laboratory
Questions?

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5 6 microbial nutrition & growth