Chapter 6: Microbial Growth

LEARNING OBJECTIVES CHECK YOUR UNDERSTANDING

6-1 Classify microbes into five groups on the Why are hyperthermophiles that grow at temperatures
basis of preferred temperature range. above 100°C seemingly limited to oceanic depths?
6-2 Identify how and why the pH of culture Other than controlling acidity, what is an advantage to
media is controlled. using phosphate salt buffers in growth media?
6-3 Explain the importance of osmotic Why might primitive civilizations have used food
pressure to microbial growth. preservation techniques that rely on osmotic pressure?
6-4 Name a use for each of the four elements If bacterial cells were given a sulfur source containing
(carbon, nitrogen, sulfur, and phosphorus) radioactive sulfur (35S) in their culture media, in what
needed in large amounts for microbial growth. molecules would the 35S be found in the cells?
6-5 Explain how microbes are classified on How would one determine whether a microbe is a strict
the basis of oxygen requirements. anaerobe?
6-6 Identify ways in which aerobes avoid Oxygen is so pervasive in the environment that it would
damage by toxic forms of oxygen. be very difficult for a microbe to always avoid physical
contact. What, therefore, is the most obvious way for a
microbe to avoid damage?
6-7 Describe the formation of biofilms and Identify a way in which pathogens find it advantageous to
their potential for causing infection. form biofilms.
6-8 Distinguish chemically defined and Could humans exist on chemically defined media, at least
complex media. under laboratory conditions?
6-9 Justify the use of each of the following: Could Louis Pasteur, in the 1800s, have grown rabies
anaerobic techniques, living host cells, candle viruses in cell culture instead of in living animals?
jars, selective and differential media,
enrichment medium.
6-10 Differentiate biosafety levels 1, 2, 3, and What BSL is your laboratory?
4.
6-11 Define colony. Can you think of any reason why a colony does not grow
to an infinite size, or at least fill the confines of the Petri
plate?
6-12 Describe how pure cultures can be Could a pure culture of bacteria be obtained by the streak
isolated by using the streak plate method. plate method if there were only one desired microbe in a
bacterial suspension of billions?
6-13 Explain how microorganisms are If the Space Station in Earth orbit suddenly ruptured, the
preserved by deep-freezing and lyophilization humans on board would die instantly from cold and the
(freeze-drying). vacuum of space. Would all the bacteria in the capsule
also be killed?

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6-14 Define bacterial growth, including Can a complex organism, such as a beetle, divide by
binary fission. binary fission?
6-15 Compare the phases of microbial If two mice started a family within a fixed enclosure, with
growth, and describe their relation to a fixed food supply, would the population curve be the
generation time. same as a bacterial growth curve?
6-16 Explain four direct methods of Why is it difficult to measure realistically the growth of a
measuring cell growth. filamentous mold isolate by the plate count method?
6-17 Differentiate direct and indirect methods Direct methods usually require an incubation time for a
of measuring cell growth. colony. Why is this not always feasible for analysis of
foods?
6-18 Explain three indirect methods of If there is no good method for analyzing a product for its
measuring cell growth. vitamin content, what is a feasible method of determining
the vitamin content?

CHAPTER SUMMARY
The Requirements for Growth (pp. 157–163)

1. The growth of a population is an increase in the number of cells.

2. The requirements for microbial growth are both physical and chemical.

Physical Requirements (pp. 157–160)

3. On the basis of preferred temperature ranges, microbes are classified as psychrophiles (cold-loving),
mesophiles (moderate-temperature–loving), and thermophiles (heat-loving).
4. The minimum growth temperature is the lowest temperature at which a species will grow, the optimum
growth temperature is the temperature at which it grows best, and the maximum growth temperature is
the highest temperature at which growth is possible.
5. Most bacteria grow best at a pH value between 6.5 and 7.5.
6. In a hypertonic solution, most microbes undergo plasmolysis; halophiles can tolerate high salt
concentrations.

Chemical Requirements (pp. 160–162)

7. All organisms require a carbon source; chemoheterotrophs use an organic molecule, and autotrophs
typically use carbon dioxide.
8. Nitrogen is needed for protein and nucleic acid synthesis. Nitrogen can be obtained from the
decomposition of proteins or from NH4+ or NO3–; a few bacteria are capable of nitrogen (N2) fixation.
9. On the basis of oxygen requirements, organisms are classified as obligate aerobes, facultative
anaerobes, obligate anaerobes, aerotolerant anaerobes, and microaerophiles.
10. Aerobes, facultative anaerobes, and aerotolerant anaerobes must have the enzymes superoxide
dismutase (2 O2– + 2 H+  ∅ O2 + H2O2) and either catalase (2 H2O2  ∅ 2 H2O + O2) or peroxidase
(H2O2 + 2 H+  ∅ 2 H2O).
11. Other chemicals required for microbial growth include sulfur, phosphorus, trace elements, and, for
some microorganisms, organic growth factors.

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Biofilms (pp. 162–163)

1. Microbes adhere to surfaces and accumulate as biofilms on solid surfaces in contact with water.
2. Biofilms form on teeth, contact lenses, and catheters.
3. Microbes in biofilms are more resistant to antibiotics than are free swimming microbes.

Culture Media (pp. 164–169)

1. A culture medium is any material prepared for the growth of bacteria in a laboratory.
2. Microbes that grow and multiply in or on a culture medium are known as a culture.
3. Agar is a common solidifying agent for a culture medium.

Chemically Defined Media (p. 165)

4. A chemically defined medium is one in which the exact chemical composition is known.

Complex Media (p. 165)

5. A complex medium is one in which the exact chemical composition varies slightly from batch to batch.

Anaerobic Growth Media and Methods (pp. 166–167)

6. Reducing media chemically remove molecular oxygen (O2) that might interfere with the growth of
anaerobes.
7. Petri plates can be incubated in an anaerobic jar, anaerobic chamber, or OxyPlate.

Special Culture Techniques (pp. 167–168)

8. Some parasitic and fastidious bacteria must be cultured in living animals or in cell cultures.
9. CO2 incubators or candle jars are used to grow bacteria that require an increased CO2 concentration.
10. Procedures and equipment to minimize exposure to pathogenic microorganisms are designated as
biosafety levels 1 through 4.

Selective and Differential Media (pp. 168–169)

11. By inhibiting unwanted organisms with salts, dyes, or other chemicals, selective media allow growth
of only the desired microbes.
12. Differential media are used to distinguish different organisms.

Enrichment Culture (p. 169)

13. An enrichment culture is used to encourage the growth of a particular microorganism in a mixed
culture.

Obtaining Pure Cultures (pp. 169–170)

1. A colony is a visible mass of microbial cells that theoretically arose from one cell.

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2. Pure cultures are usually obtained by the streak plate method.

Preserving Bacterial Cultures (p. 170)

1.Microbes can be preserved for long periods of time by deep-freezing or lyophilization (freeze-drying).

The Growth of Bacterial Cultures (pp. 171–179)

Bacterial Division (p. 171)

1. The normal reproductive method of bacteria is binary fission, in which a single cell divides into two
identical cells.
2. Some bacteria reproduce by budding, aerial spore formation, or fragmentation.

Generation Time (p. 171)

3.The time required for a cell to divide or a population to double is known as the generation time.

Logarithmic Representation of Bacterial Populations (pp. 171–172)

4.Bacterial division occurs according to a logarithmic progression (two cells, four cells, eight cells, and so
on).

Phases of Growth (pp. 172–174)

5. During the lag phase, there is little or no change in the number of cells, but metabolic activity is high.
6. During the log phase, the bacteria multiply at the fastest rate possible under the conditions provided.
7. During the stationary phase, there is an equilibrium between cell division and death.
8. During the death phase, the number of deaths exceeds the number of new cells formed.

Direct Measurement of Microbial Growth (pp. 174–178)

9. A standard plate count reflects the number of viable microbes and assumes that each bacterium grows
into a single colony; plate counts are reported as number of colony-forming units (CFU).
10. A plate count may be done by either the pour plate method or the spread plate method.
11. In filtration, bacteria are retained on the surface of a membrane filter and then transferred to a culture
medium to grow and subsequently be counted.
12. The most probable number (MPN) method can be used for microbes that will grow in a liquid medium;
it is a statistical estimation.
13. In a direct microscopic count, the microbes in a measured volume of a bacterial suspension are counted
with the use of a specially designed slide.

Estimating Bacterial Numbers by Indirect Methods (pp. 178–179)

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14. A spectrophotometer is used to determine turbidity by measuring the amount of light that passes
through a suspension of cells.
15. An indirect way of estimating bacterial numbers is measuring the metabolic activity of the population
(for example, acid production or oxygen consumption).
16. For filamentous organisms such as fungi, measuring dry weight is a convenient method of growth
measurement.