Growth, Survival and Death of Microorganisms

The Meaning of Growth: In unicellular organisms, growth leads to

an increase in the number of individuals making up a population or
.culture
Generation Time: The time needed for a bacterial cell to double
provided the necessary nutrients and environments for growth. It
.varies according to the type of bacteria
:The Measurement of Microbial Concentrations
.Cell Concentration- Viable cell count, turbidity measurement -1
Biomass Concentration- Dry weight of cells, nitrogen -2
.measurement
The Meaning of Death: For a microbial cell, death means the
.irreversible loss of the ability to reproduce(grow and divide)
 Bacterial Growth
The rate at which bacteria grow and divide varies dramatically between
species. Many multiply so fast that a single cell can become a visible
colony overnight, but others require weeks to reach the same stage. When
they are grown in broth culture, bacteria follow a recognised 4-step growth
process. In the initial lag phase the cells that have been added to the
culture adjust to their new environment and start to consume the available
nutrients. Once enough time has passed for these cells to replicate their
DNA and begin to divide they enter the exponential or log phase. During
this period the cells keep dividing regularly so that the number of cells
regularly doubles. Once the total cell population has reached a high level,
and nutrients have become limited, the bacteria cease dividing so rapidly
and enter the stationary phase. This stage is characterised by a fixed
concentration of cells over time. Eventually however a lack of available
nutrients, the buildup of waste products produced by living cells, and a
buildup of toxic products released by dying cells causes more cells to die
than are produced by division and the overall concentration of cells drops.
This final stage is called the decline phase.
 
Antimicrobial Agents
Definitions
A- Biocide
B- Bacteriostatic
C- Bactericidal
D- Sterilization
E- Disinfectants
F- Septic
G- Antiseptic
H- Aseptic
I- Preservation
J- Antibiotics
.Damage to DNA -1
Protein denaturation -2
.Disruption of cell membrane or wall -3
.Removal of free sulfhydryl group -4
.Chemical antagonism -5

Types of Antimicrobial Agents

A- Physical Agents
.Heat __ Dry: Hot air oven, Flaming -1
.Moist: Autoclave, Boiling
.Radiation__UV and Ionizing Radiations -2
Filtration -3
-B
.Alcohols__ Ethanol, Isopropanol -1
.Aldehydes__ Gluteraldehyde -2
.Biguanides__ Chlorhexidine -3
.Bisphenols -4
.Halogen-Releasing Agents__ Chlorhexidine -5
.Heavy Metal Derivatives__ Silver sulfadiazine -6
.Peroxygenes__ H2O2 -7
.Phenols -8
.Quaternary Ammonium Compounds -9
Vapor Phase Sterilants__ Ethylene oxide, H2O2, -10
.Formaldehyde
.The process of propagating organisms in the laboratory
Requirements for Growth
A- Nutrients
.The elements of the organic mater: C, H, O,N, S, P -1
Ions required for energetics and catalysis: K, Na, Fe, Mg, Ca, Cl to facilitate -2
enzymatic catalysis and to maintain chemical gradients across the cell membrane
and to allow macromolecular synthesis. Microorganisms vary widely in their
.nutritional demands and their sources of metabolic energy
.Sources of metabolic energy__ Fermentation, Respiration, Photosynthesis -3

Nutrition
Carbon source : Autotrophs and Heterotrophes. 4- Phosphorus source: -1
PO4
Nitrogen source: NO3, NO2, N2, NH4, R_NH2. 5- Growth factors -2
B- Physical Environments Requirements for
Cultivation of Microorganisms
a- Temperature
.Psychrophiles: grow best at 15-20c -1
.Mesophiles: grow best at 30-37c -2
.Thermopiles: grow best at 50-60c -3
.Extreme thermophiles: can grow at a temperature >100c -4

b- pH
.Most microorganisms have a fairly narrow optimal pH range
.Neutrophiles: 6-8 pH -1
.Acidophiles: pH as low as 3 -2
Alkaliphiles: pH as high as 10.5 -3
C- Aeration
.Obligate aerobes -1
.Obligate anaerobes -2
.Facultative anaerobes -3
.Microaerophils: requires about 5% O2 for growth -4
The natural by products of aerobic metabolism are H2O2 and -O2 superoxide and
hydroxyl radicals( OH- ) in the presence of iron. Aerobic bacteria have the
.enzymes superoxide dismutase and catalase to get rid of these toxic products
O2+ H2O2 Fe
-
O2 + OH- +*OH
2O2 + 2H O2 + H2O2
2H2O2 2H2O + O2
D- Ionic strength and Osmotic pressure (halophilic and osmophilic).
E- CO2 requirements: Capnophils (5-10%).
F- Light for photosynthetic bacteria.
The Medium
Synthetic or chemically defined medium: Simple medium, all the components are inorganic -1
.known compounds. Used for autotrophs
Undefined complex media: Composition of media not completely known. Often made from -2
inexpensive organic material such as slaughterhouse wastes,( such as tryptone, trypticase, Beef
.extract), soybeans, yeast wastes, animal blood. Such media are used the culture of heterotrophs
Types of complex media
General purpose media: nutrient broth, nutrient agar, blood agar, chocolate agar. For both -1
.gram positive and negative bacteria
Enriched media: SS agar. Encourage growth of pathogen from specimens containing normal -2
.flora
Selective media: Allow the growth of some bacteria and prevent others. MaCconkey agar for -3
gram negative bacilli and prevent gram positive bacteria. Media can be made selective by the
.addition of chemicals, antibiotics, pH
Differential media: Distinguish between different bacteria. MaCconkey agar is both -4
. selective and differential medium(lactose & nonlactose fermenters)
Assay media -5
Growth Morphology
.In broth: Turbidity, pigment production, odor -1
Slant agar: Pigment production, odor, type of growth -2
Solid media plates: colonial morphology(small, large, entire, -3
round, convex, serrated edge,mucoid, glistening, rough colonies),
.pigment production, odor

Preservation of Bacteria
:Bacteria can be preserved by
.Agar slant covered by sterile liquid paraffin -1
.Lyophilization(freeze drying) -2
Microscopic examination: Gram stain, Acid fast stain, Dark -1
.field microscopy, Phase contrast
Culture: Use streak plate to culture sample on appropriate -2
media to get pure culture. Use in addition selective media for the
.isolation of the suspected organism
Identification: use staining, differential media, serology and or -3
.gene probe for identification of the isolated microbe
.Invitro antibiotic sensitivity test -4
Epidemiology in case of outbreak to determine the source of -5
.infection
.Pure Culture : Contain one type of bacteria. Present in lab only
Mixed Culture: Contains more than one type of bacteria. Present
.in the environment
It denotes the population of microorganisms that inhibits the
skin and mucous membranes of healthy normal person. It is
.doubtful whether a normal viral flora exists in humans
:The following body organs have normal flora
Skin, Mouth, Upper Respiratory Tract, Intestinal tract(except
Esophagus, Stomach and Small Intestine), Urethra, Vagina,
.Eye, Conjunctiva
The presence of normal flora help protect against infections
by pathogens which explain infection of gastrointestinal tract
.after orally taking antibiotics(antibiotic diarrhea)