![Page 1: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/1.jpg)
1
Chapter 5
Environmental Influences and Control of Microbial Growth
![Page 2: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/2.jpg)
2
Chapter Overview
● How the environment limits growth
● The microbial response to temperature
● How microbes cope with pressure
● The microbial response to changes in: water activity, salt concentrations, pH, and oxygen
● Hungry microbes
● The control of microbes:
- Physical, chemical, and biological
![Page 3: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/3.jpg)
3
Introduction
Microbes have both the fastest and the slowest growth rates of known organisms.
Some hot-springs bacteria can double in as little as 10 minutes, whereas deep-seas sediment microbes may take as long as 100 years.
These differences are determined by nutrition and niche-specific physical parameters like temperature and pH.
![Page 4: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/4.jpg)
4
Environmental Influences and Control of Microbial Growth
Lactobacillus plantarumpH 6.5
Lactobacillus plantarumpH 3
![Page 5: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/5.jpg)
5
Environmental Limits of Microbial Growth
“Normal” growth conditions
- Sea-level; temperature 20–40oC; neutral pH; 0.9% salt; and ample nutrients
Any ecological niche outside this window is called “extreme,” and organisms inhabiting them are called extremophiles.
Figure 1.1
![Page 6: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/6.jpg)
6
Global approaches used to study gene expression allow us to view how organisms respond to changes in their environment.
- DNA microarrays assess which RNAs (Transcripts) are made in a given organism under a given condition.
- Two-dimensional protein gels achieve separation of proteins based on differences in each protein’s isoelectric point (first dimension) and molecular weight (second dimension).
Genomic and Proteomic approaches to study gene expressions
![Page 7: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/7.jpg)
7
Response to environmental stress: Global analysis: Genes and proteins expressed under two different growth conditions.
The microarrays contain all genes on a genome attached to a slide. Organisms grown under two growth conditions. Transcripts (RNAs) are converted to cDNA and hybridized to the slide. Red indicate gene expressed under one conditions and Green under different conditions. Yellow shows expression under both conditions.
The 2-D gel image shows protein expressed under 2 pH conditions
![Page 8: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/8.jpg)
8
![Page 9: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/9.jpg)
9
Effect of Temperature
• Changes in temperature impact every aspect of microbial physiology.
• Each organism has an “optimum, minimum and maximum” temperatures that define its growth limits. These are cardinal temperatures
![Page 10: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/10.jpg)
10
Temperature Optimum
Microorganisms can be classified by their growth temperature:
- Psychrophiles ~ 0–20oC (15oC)- Mesophiles ~ 15–45oC (37 oC)- Thermophiles ~ 40–80oC (60 oC)- Hyperthermophiles ~ 65–121oC (90oC)
![Page 11: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/11.jpg)
11
![Page 12: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/12.jpg)
12
Psychrophiles can be found in Icebergs
Bacterium from South Polar snow
Thermus aquatics, Yellowstone
Yellowstone hotspring Methanocaldococcus jannaschii, 780 C and 30 psi. Isolated from 2600 m deep “white smoker” chimney
![Page 13: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/13.jpg)
13
How do microbes deal with cold?
• Cytoplasmic membrane contain high-level of unsaturated fatty acids allowing their membranes to be more flexible in cold.
• Contain antifreeze proteins and cryoprotectants such as trehalose
• Enzymes from psychrophiles are useful for biotechnology such a bioremediation and for biochemical reactions at low temperature
Trehalose
![Page 14: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/14.jpg)
14
Cytoplasmic membrane contain high-level of unsaturated fatty acids allowing their membranes to be more flexible in cold.
![Page 15: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/15.jpg)
15
How do microbes deal with heat-Shock?
• Rapid temperature changes experienced by microbes show the expression of certain genes.
• The proteins, chaperones help maintain protein shape (3-D structure).
• Specialized DNA binding proteins protect DNA from denaturation.
• Membranes have high levels of saturated fatty acids or hydrocarbons. Some archaea have single lipid layer in membrane as opposed to bilayer
![Page 16: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/16.jpg)
16
Archaeal Lipids Extreme temperature archaea
![Page 17: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/17.jpg)
17
Variations in Pressure
Barophiles or piezophiles are organisms adapted to grow at very high pressures.
- Up to 1,000 atm (101 MPa, or 14,000 psi)
Barotolerant organisms grow well over the range of 1–50 MPa, but their growth falls off thereafter.
Note that many barophiles are also psychrophiles because the average temperature at the ocean floor is 2oC.
![Page 18: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/18.jpg)
18
Figure 5.5
Figure 5.6
![Page 19: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/19.jpg)
19
Changes in Water Activity
Water activity (aw) is a measure of how much water is available for use.
Osmolarity is a measure of the number of solute molecules in a solution and is inversely related to aw.
Aquaporins are membrane-channel proteins that allow water to traverse the membrane much faster than by diffusion.
- Help protect the cell from osmotic stress
![Page 20: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/20.jpg)
20
Minimizing Osmotic Stress
In addition to moving water, microbes have at least two mechanisms to minimize osmotic stress:
- In hypertonic media, bacteria protect their internal water by synthesizing or importing compatible solutes (e.g.: proline or K+)
- In hypotonic media, pressure-sensitive or mechanosensitive channels can be used to leak solutes out of the cell.
![Page 21: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/21.jpg)
21
Water Activity (aw)
aw = Vapor Pressure of a Solution
Vapor Pressure of Pure water
aw of solutions ranges from 0-1.
aw of pure water is 1 (100% humidity).
![Page 22: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/22.jpg)
22
Changes in Salt Concentrations
Halophiles require high salt concentrations.
- From 2–4 M NaCl (10–20% NaCl)
- For comparison, seawater is ~ 3.5% NaCl
![Page 23: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/23.jpg)
23
Changes in pH
Figure 5.11
![Page 24: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/24.jpg)
24
Changes in pH
All enzyme activities exhibit optima, minima, and maxima with regard to pH.
Bacteria regulate internal pH.- When environment is in a similar pH range
Weak acids can pass through membranes.- Disrupt cell pH homeostasis, and thus will kill cells- This phenomenon is used to preserve foods.
![Page 25: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/25.jpg)
25
Changes in pH
Three classes of organisms are differentiated by the pH of their growth range:
- Neutralophiles grow at pH 5–8.
- Include most pathogens
- Acidophiles grow at pH 0–5.
- Are often chemoautotrophs
- Alkaliphiles grow at pH 9–11.
- Typically found in soda lakes
![Page 26: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/26.jpg)
26
The cyanobacterium Spirulina has high concentrations of carotene, giving it a distinct pink color.
- It is also a major food for the famous pink flamingo.
Figure 5.15
![Page 27: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/27.jpg)
27
pH HomeostasisWhen cells are placed in low pH conditions,
protons can enter the cell and lower internal pH to lethal levels.
Microbes can prevent the unwanted influx of protons by exchanging extracellular K+ for intracellular H+ when the internal pH becomes too low.
Under extremely alkaline conditions, the cells can use the Na+/H+ antiporter to bring protons into the cell in exchange for expelling Na+.
![Page 28: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/28.jpg)
28
Figure 5.17
![Page 29: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/29.jpg)
29
Microbial Responses to Oxygen
Strict aerobes can only grow in oxygen.
Microaerophiles grow only at lower O2 levels.
Strict anaerobes die in the least bit of oxygen.
Facultative anaerobes can live with or without oxygen.
Aerotolerant anaerobes grow in oxygen while retaining a fermentation-based metabolism.
![Page 30: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/30.jpg)
30
Figure 5.19
Oxygen-related growth zones in a standing test tube
![Page 31: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/31.jpg)
31
Generation and destruction of reactive oxygen species (ROS)
Figure 5.20
![Page 32: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/32.jpg)
32
Culturing Anaerobes in the Lab
Three oxygen-removing techniques are used today:1. Special reducing agents (thioglycolate) or enzyme systems (Oxyrase) can be added to ordinary liquid media.2. An anaerobe jar3. An anaerobic chamber with glove ports
- O2 is removed by vacuum and replaced with N2 and CO2.
![Page 33: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/33.jpg)
33
Figure 5.21
![Page 34: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/34.jpg)
34
Microbial Response to Starvation
Starvation is a stress that can elicit a “starvation response” in many microbes.- Enzymes are produced to increase the efficiency of nutrient gathering and to protect cell macromolecules from damage.
This response is usually triggered by the accumulation of small signal molecules such as cAMP or guanosine tetraphosphate.
![Page 35: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/35.jpg)
35
Microbial Response to Starvation
Some organisms growing on nutrient-limited agar can even form colonies with intricate geometrical shapes that help the population cope, in some unknown way, to food stress.
Figure 5.22
![Page 36: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/36.jpg)
36
Oligotrophic Bacteria
In natural ecosystems, most microbes appear to be oligotrophs, organisms with a high rate of growth at low solute concentrations.- Indeed, they require low nutrient levels to survive.
Some oligotrophic bacteria have thin extensions of their membrane and cell wall called prothecae (stalks).- These expand the surface area of the cell and increase nutrient-transport capacity.
![Page 37: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/37.jpg)
37
Humans Influence Microbial Ecosystems
Maximum diversity in an ecosystem is maintained, in part, by the different nutrient-gathering profiles of competing microbes.
Figure 5.23
![Page 38: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/38.jpg)
38
Humans Influence Microbial Ecosystems
Eutrophication is the sudden infusion of large quantities of a formerly limiting nutrient.
Figure 5.24
- It can lead to a “bloom” of microbes, which can threaten the existence of competing species.
![Page 39: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/39.jpg)
39
A variety of terms are used to describe antimicrobial control measures:
- Sterilization: Killing of all living organisms
- Disinfection: Killing or removal of pathogens from inanimate objects
- Antisepsis: Killing or removal of pathogens from the surface of living tissues
- Sanitation: Reducing the microbial population to safe levels
Control of Microbes
![Page 40: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/40.jpg)
40
Microbes die at a logarithmic rate.
Decimal reduction time (D value) is the length of time it takes an agent or a condition to kill 90% of the population.
Figure 5.25
![Page 41: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/41.jpg)
41
High temperature
- Moist heat is more effective than dry heat.
- Boiling water (100oC) kills most cells.
- Killing spores and thermophiles usually requires a combination of high pressure and temperature.
- Steam autoclave
- 121oC at 15 psi for 20 minutes
Physical Agents
![Page 42: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/42.jpg)
42
Figure 5.26
![Page 43: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/43.jpg)
43
Pasteurization
- Many different time and temperature combinations can be used.
- LTLT (low-temperature/long-time)
- 63oC for 30 minutes
- HTST (high-temperature/short-time)
- 72oC for 15 seconds
- Both processes kill Coxiella burnetii, the causative agent of Q fever.
Physical Agents
![Page 44: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/44.jpg)
44
Cold
- Low temperatures slow down growth and preserve strains.
- Refrigeration temperatures (4–8oC) are used for food preservation.
- For long-term storage of cultures
- Placing solutions in glycerol at –70oC
- Lyophilization or freeze-drying
Physical Agents
![Page 45: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/45.jpg)
45
Filtration
- Micropore filters with pore sizes of 0.2 m can remove microbial cells, but not viruses, from solutions.
Physical Agents
Figure 5.27
![Page 46: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/46.jpg)
46
Air can also be sterilized by filtration.
Laminar flow biological safety cabinets force air through HEPA filters.
Figure 5.28
![Page 47: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/47.jpg)
47
Irradiation
- Ultraviolet light- Has poor penetrating power - Used only for surface sterilization
- Gamma rays, electron beams, and X-rays- Have high penetrating power- Used to irradiate foods and other heat- sensitive items
Physical Agents
![Page 48: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/48.jpg)
48
A number of factors influence the efficacy of a given chemical agent, including:
- The presence of organic matter
- The kinds of organisms present
- Corrosiveness
- Stability, odor, and surface tension
Chemical Agents
![Page 49: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/49.jpg)
49
The phenol coefficient test compares the effectiveness of disinfectants.
The Phenol Coefficient
Table 5.3
![Page 50: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/50.jpg)
50
These include:
- Ethanol
- Iodine (Wescodyne and Betadine)
- Chlorine
All of the above damage proteins, lipids, and DNA.
- Are used to reduce or eliminate microbial content from objects
Commercial Disinfectants and Antiseptics
![Page 51: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/51.jpg)
51
Figure 5.30
![Page 52: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/52.jpg)
52
Antibiotics are chemical compounds synthesized by one microbe that kill or inhibit the growth of other microbial species.
Penicillin mimics part of the bacterial cell wall.
- Prevents cell wall formation and is bactericidal
Antibiotics
Figure 5.31
![Page 53: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/53.jpg)
53
Figure 5.32
Effect of ampicillin (a penicillin derivative) on E. coli
![Page 54: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/54.jpg)
54
Biocontrol is the use of one microbe to control the growth of another.- Probiotics contain certain microbes that, when ingested, aim to restore balance to intestinal flora.- Lactobacillus and Bifidobacterium- Phage therapy aims to treat infectious diseases with a virus targeted to the pathogen.- A possible alternative to antibiotics in the face of rising antibiotic resistance
Biological Control of Microbes
![Page 55: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/55.jpg)
55
Chapter Summary● Global analysis of genes and proteins allows us to
study how microbes react to environmental changes.
● Microbes are classified by growth temperature:- Psychrophiles, mesophiles, and thermophiles
● Barophiles can grow at very high pressures. ● Halophiles require high salt concentrations.● Microbes are classified by pH range:
- Acidophiles, neutralophiles, and alkaliphiles
● Microbes are classified by their O2 requirements:- Aerobes, facultative, microaerophiles, and anaerobes
![Page 56: 1 Chapter 5 Environmental Influences and Control of Microbial Growth](https://reader035.vdocument.in/reader035/viewer/2022070407/56649e185503460f94b048ac/html5/thumbnails/56.jpg)
56
Chapter Summary● Cells treated with antimicrobials die at a logarithmic
rate.
● Physical agents used to control microbes include:
- Autoclaving, pasteurization, refrigeration, filtration, and irradiation
● Chemical agents used to control microbes include:
- Antiseptics and disinfectants
● Antibiotics selectively control microbial growth.
● Biological control of microbes includes the use of probiotics and phage therapy.