Chapter 7

Elements of:
Microbial Nutrition Environmental Factors :

Microbial Growth

Microbial Nutrition
Acquisition of nutrients for utilization Essential nutrients: Macronutrients (C, H2 and O2)

Roles in cell structure and metabolism

Roles in enzyme function and protein structure

Micronutrients (Trace elements) e.g. Mn, Zn Ni)

Bacteria are composed of: Water 70% (inorganic) Protein 15% (organic) Others include: nucleic acids, carbohydrates, lipids, etc About 96% of the cell is made of (CHONPS)
Growth factors are supplementary essential nutrients not

synthesized by an organism e.g. amino acids, vitamins

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Nutritional Types
Based on carbon source: Autotrophs (self-feeders) Use inorganic C as source e.g. CO2 Heterotrophs Depends on other life forms Use organic C as source e.g. sugars Based on energy source: Phototrophs Photosynthetic (sunlight) Chemotroph Gain energy from chemical compounds
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Nutritional types conti

Photoautotrophs (primary producers)
Energy from sunlight into chemical energy Water + Carbon dioxide = Food + Oxygen

E.g. Algae, plants, some bacteria

Chemoautotrophs (Two types)

Chemoorganic autotrophs

Derives their energy from organic compounds and their carbon source from inorganic compounds

Lithoautotrophs

Use neither sunlight nor organics nutrients but inorganic minerals. Recycling of inorganic nutrients E.g. Methanogens (methane producing acheae) 4H2 + CO2 CH4 + 2H2O 4

Heterotrophs*
Most microbes in this category referred to as chemoherotrophs*:
Derive both carbon and energy from organic compounds

Two main groups:

Saprobic

decomposers of plant litter, animal matter, and dead microbes

Parasitic

Live in or on the body of a host

Representation of a saprobe and its mode of action.

Fig. 7.2 Extracellular digestion in a saprobe with a cell wall.

Transport mechanisms for nutrient absorption

Osmosis

Diffusion
Facilitated diffusion Active transport

Endocytosis Phagocytosis (solids) formation of vacuole Pinocytosis (liquids): formation of vesicle

Osmosis
Diffusion of water through a permeable but selective

membrane Water moves toward the higher solute concentrated areas

Type of solutions:
Isotonic: same concentration Hypotonic: lower concentration

Hypertonic: higher concentration

Solute + Solvent = Solution

Representation of the osmosis process.

Fig. 7.3 Osmosis, the diffusion of water through a selectively permeable membrane

Cells with- and without cell walls, and their responses to different osmotic conditions (isotonic, hypotonic, hypertonic).

Fig. 7.4 Cell responses to solutions of differing osmotic content.

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Diffusion
Net movement of molecules from a high

concentrated area to a low concentrated area

No energy is expended (passive)

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A cube of sugar will diffuse from a concentrated area into a more dilute region, until an equilibrium is reached.

Fig. 7.5 Diffusion of molecules in aqueous solutions

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Facilitated diffusion
Transport of polar molecules and ions across the

membrane (due to complexity of cell membranes)

No energy is expended (passive)

Facilitated by carrier proteins which bind with the

molecules.
Characteristics of carrier proteins: Specificity: not all molecules accepted) Saturation: rate depends on # molecules Competition: some molecules have higher affinity
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Representation of the facilitated diffusion process.

Fig. 7.6 Facilitated diffusion

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Active transport
Transport of molecules against a gradient Requires energy (active) Use of specific membrane proteins (permeases and

protein pumps)
transport sugars, amino acids, organic acids,

phosphates and metal ions.

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Endocytosis
(Eating and drinking of cells) Requires energy (active) Types of endocytosis: Phagocytosis (solids) Pinocytosis (liquids)

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Example of the permease, group translocation, and endocytosis processes.

Fig. 7.7 Active transport

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Environmental factors that influence microbes

Temperature
Gas pH Osmotic pressure Miscellaneous Microbial association Microbes and humans

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Temperature
For optimal growth and metabolism
Psychrophile 0 to 15 C Mesophile- 20 to 40 C Thermophile- 45 to 80 C

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Growth and metabolism of different ecological groups based on ideal temperatures.

Fig. 7.8 Ecological groups by temperature

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Gas
Two gases that most influence microbial growth Oxygen Carbon dioxide

Three categories of bacteria Obligate aerobe Facultative anaerobe Obligate anaerobe

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Obligate aerobe
Requires oxygen for metabolism
Possess enzymes that can neutralize the toxic oxygen

metabolites (O, O2-, H2O2, OH-)

Superoxide dismutase and catalase 2O2- + 2H H2O2

H2O2 + O2 2H2O + O2

E.gs. Fungi, Protozoa and Bacteria (Micrococcus and Bacillus spp)

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Facultative anaerobe

Does not require oxygen for metabolism, but can grow in

its presence During minus oxygen states, anaerobic respiration or fermentation occurs Possess superoxide dismutase and catalase Most bacterial pathogens in this group.
E.g.s

Gram negative intestinal bacteria

Staphylococcus spp.

Microaerophiles:
Require minute amounts of O2 (lower than atmospheric concentration for

metabolism (live in water, soil, human body

Aerotolerant anaerobes: e.g. lactobacillus and streptococci

Do not utilize O2 but can survive in its presence Possess alternate mechanisms for removing toxic O2 products e.g.

Manganese ions.
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Facultative anaerobes
Does not require oxygen for metabolism, but can grow

in its presence During minus oxygen states, anaerobic respiration or fermentation occurs Possess superoxide dismutase and catalase
Ex. Gram negative pathogens Neisseria (gonorrhea, meningitis,) Brucella (undulant fever) Streptococcus pneumoniae require 3 10% CO2.

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Obligate anaerobes
Cannot use oxygen for metabolism
Do not possess superoxide dismutase and catalase The presence of oxygen is toxic to the cell The oral cavity and large intestines present anaerobic

conditions for some bacteria.

Clostridium is an anaerobe that infects humans

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Anaerobes must grow in an oxygen minus environment, because toxic oxygen metabolites cannot be neutralized.

Fig. 7.10 Culturing technique for anaerobes

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Thioglycollate broth enables the identification of aerobes, facultative anaerobes, and obligate anaerobes.

1. Aerobic (Pseudomonas aeruginosa) 2&3 Facultative Staphylococcus aureus and E. coli 4. Obligate anaerobe ( Clostridium butyricum)

Fig. 7.11 Use of thioglycollate broth to demonstrate oxygen requirements. Depends on relative utilization of oxygen for growth
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pH
pH:- measure of Hydrogen ion concentration
Most Cells grow best between pH 6-8 Exceptions : Acidophiles (pH 0, 1, 2) Euglena (alga) Thermoplasma (acheae) Alkalinophiles (pH 10).

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Osmotic pressure
Most microbes grow under isotonic and hypotonic

conditions, however some thrive in hypertonic conditions : Halophiles Requires high salt concentrations
Ex. Halobacterium

Facultative halophiles Can survive high salt conditions but is not required E.g. Staphylococcus aureus

Miscellaneous
Radiation- withstand UV, infrared Barophiles withstand high pressures Spores and cysts- can survive dry habitats
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Ecological associations
Symbiosis (tic):
Two organisms that live in close nutritional relationship Mutualism both organism benefit Commensalism one organisms benefits Parasitism host/microbe relationship

Non-symbiotic:
Organisms are free-living, and do not rely on each other

for survival
Synergism shared metabolism Antagonism- competition between microorganisms In humans, microbial associations could be commensalism, parasitic or synergistic. E. coli produces vitamin K for host
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Microbial Growth
Binary fission Division of the bacterial cell (reproduction) Generation time Time required for a complete division cycle (doubling) Length of the generation time is a measure of the growth rate Exponentials are used to define the numbers of bacteria after growth Growth curve: 4 phases: Lag, Log, Stationary and Death phases Enumeration of bacteria: Counting
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Representation of how a single bacterium doubles after a complete division, and how this can be plotted using exponentials.

Fig. 7.14 The mathematics of population growth

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Growth curve
Lag phase Cells are adjusting, enlarging, and synthesizing critical proteins and metabolites Not doubling at their maximum growth rate Log phase Maximum exponential growth rate of cell division Adequate nutrients Favorable environment

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Growth curve conti

Stationary phase Survival mode depletion in nutrients, released waste can inhibit growth Number of cells that stop dividing = # of cells dividing Death phase Exponential death of cells due to lack of nutrients
In industry, a chemostat is used to provide a continuous

supply of nutrients (Vitamin and antibiotic production), Thus the death phase is never achieved.

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The four main phases of growth in a bacterial culture.

Fig. 7.15 The growth curve in a bacterial culture.

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Enumeration of bacteria
Bacteria are enumerated by the following methods:
Turbidity Direct cell count Automated devices Coulter counter Flow cytometer Real-time Polymerase Chain Reaction (PCR)

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The greater the turbidity, the larger the population size.

Fig. 7.16 Turbidity measurements as indicators of growth

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The direct cell method counts the total dead and live cells in a special microscopic slide containing a premeasured grid.

Fig. 7.17 Direct microscopic count of bacteria.

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A Coulter counter uses an electronic sensor to detect and count the number of cells.

Fig. 7.18 Coulter counter

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