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DISINFECTION

Dfinitions
Bactricide / Bactriostatique
Mort / inhibition de croissance

Dsinfection
Toutes actions antimicrobiennes

Antisepsie
Dsinfection sur organismes vivants

Strilisation
Destruction de lensemble des microorganismes

Spectre antimicrobien

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Disinfectants

Antimicrobial agents

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Types of disinfectants

Water disinfectants
Chlorine
Chloramines (monochloramine)
Chlorine dioxide
Ozone
Peroxide
Iodine
Metals (Cu, Ag)

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Other treatments
Temperature
Filtration
UV
Electrochemistry
Photocatalysis

Source: The handbook of water and wastewater microbiology Academic Press

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Disinfectant activity
Destroy or inactivate MO
Wide range of activity (on many MO species)
Persistent
Limited toxicity for human and environment

Disinfection depends on use

Drinking water (chlorine, monochloramine)
Wastewater (chlorine, ozone)
Industrial water (temperature, chlorine for cooling
tower)
Water for pharmaceutical industry (need to
eliminate all traces of MO)

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Chlorine
Highly efficient in water treatment
Problems due to generation of dangerous byproducts (toxic or carcinogenic) ex: THM
Debate around the use of chlorine
Some countries does not use any treatment for
potable water
Not always sufficient to eliminate all MO

Chlorine concentration

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Reaction of chlorine in water

Mode of chlorine treatment

Shock Chlorination (50-100 ppm, contact of at least 6
hours)

Continuous Chlorination for recurring bacterial

contamination problems a measurable amount of free
residual chlorine (0.5 ppm = mg/l)

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Disinfectant efficiency

Inactivation kinetics
1 log inactivation 90% killed
2 log inactivation 99% killed
3 log inactivation 99,9% killed

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Ct
Ct = concentration x time
Time influence inactivation efficiency

Ct99%= Ct for 2 log reduction

Source CDC

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How to measure efficiency?

Cultivation
Viability
Presence of cells, DNA

Survival curves
2 lag phase before efficiency
Differences between strains
Selection of resistance (ex antibiotics)

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Factors influencing efficiency

Stability (efficiency decreases with time)
Concentration (modification of effect)
pH (ex chlorine at acidic pH is more efficient)
Temperature
Interfering compounds (organic mainly)
MO factors

pH and chlorine
pH modify active species
Hypochlorous acid (HOCl) is more potent than
hypochlorite ions (OCl)

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pH
Ammonium compounds more potent at basic pH
Chorine dioxide is equally active between 6 and 10

Mode of action Disinfectants

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Oxidation
O2 is not toxic by itself
Oxygen derivatives (named ROS for oxygen
reactive species) might be toxic for MO

HO. being the more toxic

Mode of action of oxidant

Oxidation of host molecules leads to inactivation
Ex DNA, proteins, lipids

Oxidation of lipids in the membrane leads to

internal efflux
Sublethal concentration lead to respiration inhibition
(loss of energy production)

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Superoxide O2Hydrogen peroxide H2O2

Hydroxyl radical HO.
Singlet oxygen O
Hypochlorous acid (bleach) react with sulphurous
amino acids

DNA Damage causes by oxidants

DNA damages
Only HO. can oxidize both base and ribose moiety of
DNA
Guanine is the main target (lower reduction potential) >
production of 8hydroxy guanine > able to base pair
with adenine (highly mutagenic)
Oxidation of ribose induces DNA strand break

Bacteria possess enzymes to repair DNA

Recombination genes (rec)

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Protein and lipid damages

Amino acids oxidation
Cysteine is the more sensitive > formation of disulphide
bond
Most protein are not targeted by physiological doses
of oxidants

Lipid peroxidation (mainly in eukaryotes, need of

polyinsaturated lipids)

Damage of enzyme ironsulphur

clusters
Inactivation of enzyme
Production of iron

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Resistance to disinfectants

Resistance to disinfection
Differences between species
Differences between strains
Selection of resistance (ex antibiotics)

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Mean of resistance
Cell envelope (differences between MO)
Cysts and spores
Protection within a host (Legionella amoebae)
Efflux pumps
Degradation of disinfectants (ex H2O2)
Aggregates, biofilms
Adaptation (phenotypic modification)
Mutation (genetic modification)
Repair mechanism

Mcanismes de rsistance aux antibiotiques

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