(micro bio) microbial nutrition
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MICROBIALNUTRITION AND
GROWTHI reserve all apologies for all copyright material used here. These are mostly
3rdhand materials from the web. This presentation is purely for educationalpurposes. If you believe that your copyright material is not geared to educate,
please inform me ([email protected]). I will always respect your
beliefs. *Please note that I disabled all animations to prevent issues after downloading*
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MICROBIAL NUTRITION
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Biochemical Components of Cells
Water: 80 % of TOTAL BIOMASS
Dry weight
Protein 40-70 %
Nucleic acid 13-34%
Lipid 10-15 %
Also monomers, intermediates and
inorganic ions
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Macronutrients
These are used to synthesize biomolecules (C, O,H, N, S, P) and to regulate many important
metabolic and structural functions (K+, Ca2+, Mg2+,
Fe2+)
proteins, carbohydrates, lipids and nucleic acids are
primarily made up of (COHNSP)
ions have variable roles (eg. K+ is needed to activate
enzymes during photosynthesis; Ca2+ is responsible for the
heat resistance of bacterial endospores; Mg2+ is a cofactor
for many enzymes; Fe2+/3+ is an important component of
electron transport systems)
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Micronutrients
These are needed in trace quantities that even very
slight contaminants in aqueous environments are
enough to meet the demands of microbes.
Trace elements include Ni, Mo, Co, Cu, Mn, and ZnGrowth factors such as vitamins and some enzymes are
also essential but are needed only in trace quantities.
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NUTRITIONAL TYPES
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NUTRIENT UPTAKE
Passive Transport does not require energy; substancesexist in a gradient and move from areas of higherconcentration towards areas of lower concentration diffusion osmosis (diffusion of water) facilitated diffusion (permeases)
Active Transport requires energy and carrier proteins;gradient independent
active transport (symport, antiport) group translocation (transported molecule chemically
altered) bulk transport (endocytosis, exocytosis, pinocytosis)
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DIFF
U
SIONACRO
SSAM
E MBR
AN
E
PASSIVE
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TONICITY
PASSIVE
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FACILITATED DIFFUSION
-utilizes channel and carrierproteins collectively called as
permeases
PASSIVE
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A
CTIVETRA
NSPORT
INPROCARYOTES
ACTIVE
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ACTIVE TRANSPORT USUALLY UTILIZES
TRANSPORT PROTEINS
ACTIVE
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Group Translocation
5 proteins involved.
transported substance (glucose) is chemically
altered (glucose-6-P) during transportation
ACTIVE
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ACTIVE
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Group Translocation
ACTIVE
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SIDEROPHORES
ACTIVE
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ENDOCYTOSIS
ACTIVE
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MICROBIAL
GROWTH
Microbes especially bacteria
divide asexually via binary
fission.
Generation Time
- the time required for parent
cell to form two new daughter
cells.
Microbial growth is always
measured by number, not by
biomass.
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MICROBIAL GROWTH CURVE
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Nf= Final population
Ni = Initial population
2n
= # cells in generation
n= generation number
Nf = (Ni)2n
Calculating Growth of Cells
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Doubling times for some common bacteria
under optimal conditions of growth
Bacterium Medium Generation Time(minutes)
Escherichia coli Glucose-salts 17
Bacillus megaterium Sucrose-salts 25
Streptococcus lactis Milk 26
Streptococcus lactis Lactose broth 48
Staphylococcusaureus
Heart infusion broth 27-30
Lactobacillusacidophilus
Milk 66-87
Rhizobium japonicum Mannitol-salts-yeastextract
344-461 (6hr)
Mycobacteriumtuberculosis
Synthetic 792-932 (15hr)
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Measuring Growth: Direct Count, Serial
Dilutions, Plate Counts and Turbidity
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A. Direct Count
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Spread-plate method Pour-plate method Pour-plate method with serial
dilution
B. Viable Count
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C. Indirect Count: Turbidity Measurement
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Calculation of the Number of Bacteria per milliliter of
Culture Using Serial Dilution
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Pour plate:
made by first
adding 1.0ml ofdiluted culture to
9ml of molten
agar
Spread plate:
made by adding
0.1ml of diluted
culture to surfaceof solid medium
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Co
lonyC
ounter
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Plate as viewedunder the colony
counter grid.
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Countable number of colonies
(30 to 300 per plate)
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The Petroff-Hausser Counting Chamber
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Turbidity, or a cloudy
appearance, is an
indicator of bacterialgrowth in urine in the
tube on the left
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A Spectrophotometer: This instrument can be used to
measure bacterial growth by determining the degree of
light transmission through the culture
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STREAK
PLATETECHNIQUE
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A Streak Plate
ofSerratiamarcescens.
Note the greatly
reduced
numbers ofgrowth
/colonies in
each
successive
region.
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Types of Culture Media
Natural Media: In nature, many species ofmicroorganisms grow together in oceans, lakes, and
soil and on living or dead organic matter
Synthetic medium (Defined): A medium prepared in thelaboratory from material of precise or reasonably well-
defined composition
Complex medium (Undefined): contains reasonablyfamiliar material but varies slightly in chemical
composition from batch to batch (e.g. peptone, a
product of enzyme digestion of proteins)
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Selective, Differential, and
Enrichment Media
Selective Medium: encourages growth of someorganisms but suppresses growth of others
(e.g. antibiotics)
Differential Medium: contains a constituent thatcauses an observable change that can differentiate
one population from the other (e.g. MacConkey agar)
Enrichment Medium: contains special nutrients thatallow growth of a particular organism that might nototherwise be present in sufficient numbers to allow it
to be isolated and identified
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Bristol's Medium for Algae(mg/liter)
NaNO3
250 mg
K2HPO
475 mg
KH2PO
4175 mg
CaCl2 25 mgNaCl 25 mg
MgSO4.7H
2O 75 mg
FeCl3
0.3 mg
MnSO4.
4H2O 0.3 mg
ZnSO4.7H
2O 0.2 mg
H3BO
30.2mg
CuSO4.5H
2O 0.06 mg
DEFINED MEDIA
-all components are known
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COMPLEX MEDIA - components are unknownOfCas
einNutri
PepAcid
Hydrolyzate
Peptone/Tryptone/Yeast Extract
Bacteriological Media Ingredients
ANIMAL BASED & ANIMAL FREE VEGETABLE
PEPTONES:- Wide gamut of peptones for cell
nutrition, fermentation and media ingredients.
Peptones are low-cost growth promoting nutrients
used extensively as Dehydrated Culture media
ingredients,fermentation ingredients as animal cell
culture nutrients.
Animal Free Peptones
VegiPep S : Soy Peptone NON ANIMAL,NON GMO
VegiPep G : Peanut Peptone NON ANIMAL,
NON GMO Animal Origin Peptones
NutriPep Bacto Peptone
Nutri Pep Casein Peptone
Nutri Pep Meat PeptoneNutriPep Proteose Peptone
NutriPep Pancreatic Digest Of Casein
Nutri Pep Acid Hydrolyzate of Casein
Nutri Pep Acid Hydrolyzate of Soy
NutriPep Tryptose Nutri Pep Tryptone.
NutriPep Meat Extract Powder
Nutri Pep Yeast Extract Powderby_bryanyves
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The detection ofSalmonella
species is often complicated by
the presence of background flora
and other Enterobacteriaceae on
an agar plate. The presence of theselective agent, Tergitol 4, in
XLT-4 Agar inhibits many
organisms that can be
problematic on other plating
media. In addition, biochemical
and pH changes within themedium allow Salmonella spp.
(black colonies) to be
differentiated from organisms,
such as E. coli(yellow colonies)
and Shigella spp. (red colonies).
XLT-4 Agar
SELECTIVE MEDIA favor growth of organism of interest
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Blood Agar is a bacterial differential medium that can
distinguish normal from pathogenic bacteria based on theinteraction of sheep's blood and bacterial hemolytic enzymes.
DIFFERENTIAL MEDIA show comparative changes
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denti fic
ati on
ofurina
rytractpa
thog
enswith
differ e
ntialme
d ia(CHROM
agar )
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FACTORS AFFECTING GROWTH
Temperature
pH
Oxygen
Pressure
Salinity
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Effects of Temperature on Growth
95oF77oF40oF
Most of our plates are incubated at 37oC(98.6oF).
Conversion C to F = 1.8xC + 32
http://www.sciencemadesimple.com/conversions.html
Image: Pearson Education Inc. (2004) publishing as Benjamin Cummingsby_bryanyves
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Categories of Microbes Based on Temperature Range
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Psychrophiles (< 20 oC)
A form of psychrophile named Strain34h,
stained fluorescent blue for better viewing.
Polar regions are the usual
habitat of psychrophiles.
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Mesophiles (15-45oC)
The habitats of these organisms include soil, the human body,animals, etc. The optimal temperature of many pathogenic
mesophiles is 37 C (98 F), the normal human body
temperature. Mesophilic organisms have important uses in
food preparation especially in cheese and yogurt making and
in beer and wine making.
E. coli
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Thermophiles (45-80oC)
Thermophiles produce some of the
bright colors of Grand Prismatic
Spring, Yellowstone National Park
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Hyperthermophiles (65-110oC)
EXTREMELY HOT!!!
MostlyArchaeans believed to
be the first microbes on
earth!!! by_bryanyves
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pH
and
Growth
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Meet the Microbe!
Vibrio cholerae (Cholerabacteria) will grow
outside the body at apH of 9.0.
Acidobacterium-abundant in soils
(pH 3-5)
These unknown bacteria
were taken from the film on
the surface of sulfuric acid.
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Obligate Aerobes need oxygen to stay alive
Obligate Anaerobes die in presence of oxygen
Facultative Anaerobes not strict anaerobes; optimum
growth in anaerobic conditions but can still grow in thepresence of oxygen
Microaerophillic Bacteria require oxygen levels lowerthat that found under normal atmospheric conditions; aerobic
but optimum growth is when oxygen is minimal
Aerotolerant Anaerobes - can tolerate a small amount ofoxygen by being able to detoxify the poisonous forms of
oxygen present
Oxygen Requirements
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1: Obligate aerobic bacteria gather at top of test tube to absorb maximal amount of O2.
2: Obligate anaerobic bacteria gather at bottom to avoid oxygen.3: Facultative bacteria gather mostly at the top, since aerobic respiration is beneficial;
but as lack of oxygen does not hurt them, they can be found all along the test tube.
4: Microaerophiles gather at upper part of test tube. They require O2, but at low
concentration.
5: Aerotolerant bacteria are not affected by oxygen, and they are evenly spread along
the test tube. by_bryanyves
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OsmoticPressure
Some bacteria can be plasmolyzed by high
concentrations of solutes, which results in
the formation of plasmolysis bays
throughout the length of the cell.
These bays result from the contraction of
the cytoplasmic membrane in response to
hyperosmotic shock.
Why can you keep honey on the cupboard
for months, even years, without it spoiling?
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Salinity
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Salinity
Seawater: 3% NaCl Nonhalophile: less than 1%
Halotolerant: less than 5% Halophile: 1 to 15 % Extreme halophile: more than 15%
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Pressure
Barophiles- organisms that grow
at elevated pressure(3-1000 x air
pressure). (Found inocean depths often
in thermal vents)
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THANKS!!!