Antibiotics: mode of action

and mechanisms of resistance.

By
Senior scientist Henrik Hasman
National Food Institute-DTU
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What are antibiotics?

Originally:
Naturally occurring microbial products

Today:
Any agent used to treat infections
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Mechanisms of antibiotics

Bacteriostatic

Bactericidal
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Bacteriostatic antibiotics
Tetracyclines
Spectinomycin
Sulphonamides
Macrolides
Chloramphenicol
Trimethoprim
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Bactericidal antibiotics
Penicillins
Cephalosporins
Fluoroquinolones (Ciprofloxacin)
Glycopeptides (Vancomycin)
Monobactams
Carbapenems
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What is antimicrobial resistance I?

The ability of a microorganism to survive

at a given concentration of an antimicrobial
agent at which the normal population of the
microorganism would be killed

This is called the Epidemiological

breakpoint.
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Population distribution

Resistant population
=

60

50 Sensitive population
Number of isolates

40

30
Break point

20

10

0
0,25 0,5 1 2 4 8 16 32 64 128 256

MIC
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What is antimicrobial resistance II?

The ability of a microorganism to
survive treatment with a clinical
concentration of an antimicrobial
agent in the body.

This is called the Clinical breakpoint.

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Antibiotics: Modes of action

Inhibitors of DNA synthesis

Inhibitors of bacterial protein synthesis

Inhibitors of bacterial cell wall synthesis

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From DNA to protein

RNA polymerase

Nucleotides Ribosome
A G

T C

RNA polymerase
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Genetic characterisation to study

Spread of antimicrobial resistance
Folic acid metabolism: DNA gyrase: RNA polymerase:
Trimethoprim Naldixcin acid
Rifampicin
Narfloxacin
Sulfoamids Novobiocin
Quinolones
Protein syntesis
:
(60S inhibitorer)
PABA
Makrolider
(erythromycin)
THF Chlorampinicol
Clindamycin
Lincomycin
Streptogramins
DHF Oligosaccharides
Protein syntesis
Cell wall syntesis: (30S inhibitorer):
Cycloserine Tetracyclin
Vancomycin (glycopeptides) Spectinomycin
Bacitracin Streptomycin
Penicillins Gentamycin
Cephalosporins Tobramycin
Monobactam Kanamycin
Carbapenems Amikacin

(PABA= p-aminobenzocis acid, DHF= dihydrofolate, THF= tetrahydrofolate (essential for growth of bacteria))

Targets in the bacterium

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Bacterial growth
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Inhibition of cell wall synthesis

Penicillin
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Antibiotics: Modes of resistance

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Resistance mechanisms I
Point mutations in target genes/influx pumps
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Genetic variations/Point mutations

DNA gyrase quinolone resistance

110 120 130 140 150

S
Nal 101 TGACGTAATC GGTAAATACC ATCCCCACGG CGATTCCGCA GTGTATGACA
NalR MUT83A 101 TGACGTAATC GGTAAATACC ATCCCCACGG CGATTACGCA GTGTATGACA
NalR MUT83T 101 TGACGTAATC GGTAAATACC ATCCCCACGG CGATTTCGCA GTGTATGACA

Codon 83: TCC Ser

TTC Phe
TAC Tyr
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Resistance mechanisms II
Acquired genes
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Acquisition of resistance
E. coli/ salmonella
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Co-selection of resistance

vanA
175 kb
erm(B)

tcr
Origin tcr=Transferable copper resistance

Usage of copper or erythromycin selects for

presence of vancomycin resistance
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Antimicrobials and resistance problems

Emerging problems

Fluoroquinolones-resistant Salmonella
3rd gen. Cephalosporin-resistant Salmonella (ESBL)
Fluoroquinolone- and macrolide-resistant Campylobacter
Vancomycin resistant enterococci (VRE)
(Multiresistant E. coli)
MRSA in animals (report of high prevalence of MRSA in pigs
in the Netherlands - now also found in Danish animals).