Microbial Growth (Ch 6)

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1 Microbial Growth (Ch 6)

2 Figure 6.1 Typical growth rates of different types of microorganisms in response to temperature. Thermophiles Psychrotrophs Psychrophiles Mesophiles Hyperthermophiles

3 Applications of Microbiology 6.1 A white microbial biofilm is visible on this deep-sea hydrothermal vent. Water is being emitted through the ocean floor at temperatures above 100 C.

4 Figure 6.2 Food preservation temperatures. Temperatures in this range destroy most microbes, although lower temperatures take more time. Very slow bacterial growth. Danger zone Rapid growth of bacteria; some may produce toxins. Many bacteria survive; some may grow. Refrigerator temperatures; may allow slow growth of spoilage bacteria, very few pathogens. No significant growth below freezing.

5 Figure 6.3 The effect of the amount of food on its cooling rate in a refrigerator and its chance of spoilage. 15 cm (6 ) deep 5 cm (2 ) deep Approximate temperature range at which Bacillus cereus multiplies in rice Refrigerator air

6 Figure 6.4 Plasmolysis. Plasma membrane Cell wall H 2 O Plasma membrane Cytoplasm Cytoplasm NaCl 0.85% Cell in isotonic solution. NaCl 10% Plasmolyzed cell in hypertonic solution.

7 Table 6.1 The Effect of Oxygen on the Growth of Various Types of Bacteria

8

9 Table 6.2 A Chemically Defined Medium for Growing a Typical Chemoheterotroph, Such as Escherichia coli

10 Table 6.3 Defined Culture Medium for Leuconostoc mesenteroides

11 Table 6.4 Composition of Nutrient Agar, a Complex Medium for the Growth of Heterotrophic Bacteria

12 Simmons Citrate Agar Tryptic Soy Broth

13

14

15 Figure 6.6 A jar for cultivating anaerobic bacteria on Petri plates. Lid with O-ring gasket Clamp with clamp screw Envelope containing sodium bicarbonate and sodium borohydride Anaerobic indicator (methylene blue) Palladium catalyst pellets Petri plates

16

17 Figure 6.9 Blood agar, a differential medium containing red blood cells. Bacterial colonies Hemolysis

18 Figure 6.10 Differential medium. Uninoculated Staphylococcus epidermis Staphylococcus aureus

19 Biosafety levels

20 Figure 6.7 An anaerobic chamber. Air lock Arm ports

21 Biological Safety Cabinet. Arm ports labconco.com

22 Biological Safety Cabinet. Arm ports genomica.uaslp.mx

23 Biological Safety Cabinet. Arm ports usu.edu

24 Biosafety level 3 (BSL-3)

25 Figure 6.8 Technicians in a biosafety level 4 (BSL-4) laboratory.

26 Figure 6.11 The streak plate method for isolating pure bacterial cultures Colonies

27 Figure 6.12a Binary fission in bacteria. Cell elongates and DNA is replicated. Cell wall Plasma membrane Cell wall and plasma membrane begin to constrict. DNA (nucleoid) Cross-wall forms, completely separating the two DNA copies. Cells separate. (a) A diagram of the sequence of cell division

28 Figure 6.12b Binary fission in bacteria. DNA (nucleoid) Partially formed cross-wall Cell wall (b) A thin section of a cell of Bacillus licheniformis starting to divide 2013 Pearson Education, Inc.

29 Figure 6.13a Cell division.

30 Figure 6.13b Cell division.

31 Figure 6.14 A growth curve for an exponentially increasing population, plotted logarithmically (dashed line) and arithmetically (solid line). (1,048,576) Log 10 = 6.02 Log 10 of number of cells Log 10 = 1.51 Log 10 = 3.01 Log 10 = 4.52 (65,536) (32,768) (32) (1024) (262,144) (131,072) (524,288) Number of cells Generations

32 Figure 6.15 Understanding the Bacterial Growth Curve. Lag Phase Intense activity preparing for population growth, but no increase in population. Log Phase Logarithmic, or exponential, increase in population. The logarithmic growth in the log phase is due to reproduction by binary fission (bacteria) or mitosis (yeast). Stationary Phase Period of equilibrium; microbial deaths balance production of new cells. Death Phase Population Is decreasing at a logarithmic rate. Staphylococcus spp.

33 Figure 6.16 Serial dilutions and plate counts. 1 ml 1 ml 1 ml 1 ml 1 ml Original inoculum 9 m broth in each tube Dilutions 1:10 1:100 1:1000 1:10,000 1:100,000 1 ml 1 ml 1 ml 1 ml 1 ml Plating 1:10 1:100 1:1000 1:10,000 1:100,000 (10-1 ) (10-2 ) (10-3 ) (10-4 ) (10-5 ) Calculation: Number of colonies on plate reciprocal of dilution of sample = number of bacteria/ml (For example, if 54 colonies are on a plate of 1:1000 dilution, then the count is = 54,000 bacteria/ml in sample.)

34 Figure 6.17 Methods of preparing plates for plate counts. The pour plate method The spread plate method Inoculate empty plate. 1.0 or 0.1 ml 0.1 ml Inoculate plate containing solid medium. Add melted nutrient agar. Bacterial dilution Spread inoculum over surface evenly. Swirl to mix. Colonies grow on and in solidified medium. Colonies grow only on surface of medium.

35 Figure 6.18 Counting bacteria by filtration.

36 Figure 6.19a The most probable number (MPN) method. Volume of Inoculum for Each Set of Five Tubes (a) Most probable number (MPN) dilution series.

37 Figure 6.19b The most probable number (MPN) method. (b) MPN table.

38 Figure 6.20 Direct microscopic count of bacteria with a Petroff-Hausser cell counter. Grid with 25 large squares Cover glass Slide Bacterial suspension Cover glass Slide Bacterial suspension is added here and fills the shallow volume over the squares by capillary action. Location of squares Cross section of a cell counter. The depth under the cover glass and the area of the squares are known, so the volume of the bacterial suspension over the squares can be calculated (depth area). Microscopic count: All cells in several large squares are counted, and the numbers are averaged. The large square shown here has 14 bacterial cells. The volume of fluid over the large square is 1/1,250,000 of a milliliter. If it contains 14 cells, as shown here, then there are 14 1,250,000 = 17,500,000 cells in a milliliter.

39 Figure 6.21 Turbidity estimation of bacterial numbers. Light source Light Spectrophotometer Scattered light that does not reach detector Blank Light-sensitive detector Bacterial suspension

40 Figure 6.5 Biofilms. Clumps of bacteria adhering to surface Migrating clump of bacteria Surface Water currents

41 Applications of Microbiology 3.2 Pseudomonas aeruginosa biofilm Pearson Education, Inc.

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