Upload
taiz-university
View
4.242
Download
0
Embed Size (px)
Citation preview
12/10/2014 1
CHAPTER 12Antibiotic Mechanisms of
Action and ResistanceDiagnostic Microbiology
Medical LaboratoryFaculty of Medicine & health sciences
Dr. Hafez Alsumairy
12/10/2014 2
Objectives1. Describe the mechanism of action of the different classes of
antimicrobials.2. Describe the targets of the different antibiotic classes.3. Describe and distinguish differences between intrinsic and
acquired mechanisms of antibiotic resistance.4. Discuss the mechanisms used by microorganisms to disseminate
resistant determinants.
12/10/2014 3
Introduction • Antimicrobial agents include antiseptics, antibiotics, preservatives,
sterilants, and disinfectants; all have the capacity to kill or suppress the growth of microorganisms.• Intrinsic resistance• Passed vertically to progeny
• Acquired resistance• Result of chromosomal mutations, or by the horizontal transfer
12/10/2014 4
12/10/2014 5
inhibition of bacterial Cell wall biosynthesisPeptidoglycan biosynthesis
(1)synthesis of precursors in the cytoplasm (2)transport of lipid bound precursors across
the cytoplasmic membrane(3)insertion of glycan units into the cell wall(4)transpeptidation linking and maturation.
12/10/2014 6
Antibiotic targets and mechanisms of action
12/10/2014 7
12/10/2014 8
inhibition of bacterial Cell wall biosynthesis• β-lactam antibiotics• functions as a structural analogue of
the normal substrate acyl-D alanyl-D-alanine
• penicillin-binding proteins (PBPs)• PBPs transpeptidase/transglycosylase
• Glycopeptides• Vancomycin
12/10/2014 9
Inhibition of bacterial Cell wall biosynthesis
12/10/2014 10
12/10/2014 11
Inhibition of folate synthesis• Completely synthetic molecules • Sulfamethoxazole (SMZ)• Structural analogue para
aminobenzoic acid PABA
• Trimethoprim (TMP)• Binds to dihydrofolate reductase
• Urinary tract infections
12/10/2014 12
Interference of DNA Replication• Quinolones• Targeting topoisomerases II (DNA
gyrase) in gram-negative bacteria and IV in gram-positive bacteria• Enterobacteriaceae,
pseudomonads, and other non-Enterobacteriaceae, staphylococci, enterococci, neisseria, and streptococci species other than Streptococcus pneumoniae
12/10/2014 13
Interference of DNA transcriptionRifampin• A synthetic derivative of rifamycin
B• Mycobacterium tuberculosis• The target of rifampin is the RNA
polymerase β subunit at an allosteric site• staphylococci, enterococci,
Haemophilus spp., and S.pneumoniae
12/10/2014 14
12/10/2014 15
Interference of mRNA translation• Formylmethionyl tRNA, which
binds to the P site of the 30S ribosomal subunit• Aminoglycosides are cationic
CHO-containing molecules provides the basis for their interaction with a specific region of the 16S rRNA in the 30S.• A-binding site
12/10/2014 16
Interference of mRNA translation• Polyketide class of antibiotics• Tetracycline, doxycycline, and
minocycline.
• Tetracyclines reversibly inhibit protein synthesis by binding to 16S ribosomal RNA near the amino-acyl tRNA acceptor (A) site
12/10/2014 17
Interference of mRNA translation• Macrolides such as
erythromycin, clarithromycin, and azithromycin target the 50S subunit specifially by binding tothe peptidyltransferase cavity in the proximity of the A and P loops, near adenine 2058 of 23S rRNA
12/10/2014 18
Interference of mRNA translation• Linezolid • Inhibitors by blocking the initiation
step while macrolides and tetracyclines block peptide chain elongation.• Streptogramin
• Glycylcycline• Binding to the 30S ribosomal
subunit and blocking entry of aminoacyl tRNA molecules into the A site of the ribosome
12/10/2014 19
12/10/2014 20
12/10/2014 21
Mechanisms of antibiotic resistance
• Intrinsic• Innate (chromosomal DNA)• Transmitted to progeny vertically
• Acquired• DNA by transformation• Recombination• Extrachromosomal DNA
transmitted horizontally• Plasmids• Transposons
Intrinsic (Chromosomal) Mechanisms of Resistance1. Impermeability2. Biofilms3. Efflux4. Enzymatic Inactivation
12/10/2014 22
Intrinsic (Chromosomal) Mechanisms of Resistance1. Impermeability
A. Lipopolysaccharide (LPS)composition• Strongly negatively charged core• O-antigen side chains
B. Outer membrane proteins (Omps) called porins.• E. coli are OmpF, OmpC, and OmpE• Decreases or loss of porin synthesis
• Glycopeptide
2. Biofilms• Biofilms are sessile bacterial
communities that are irreversibly attached to a substrate and embedded in an exopolysaccharide matrix. • Single or multiple bacterial
species• Chemical and physical
12/10/2014 23
Intrinsic (Chromosomal) Mechanisms of Resistance2. Biofilms• Innate resistance factors
• Structure and physiology• Persister cells
• Induced resistance factors
3. Efflux• Efflux pumps are naturally occurring • Five major efflux pump protein1. The major facilitator subfamily (MFS), 2. The resistance nodulation cell division
subfamily (RND), • Pseudomonas aeruginosa (MexAB)
3. The small multidrug regulator subfamily (SMR),
4. The adenosine triphosphate (ATP)-binding cassette (ABC) family
5. The multidrug and toxic effects (MATE) family.
12/10/2014 24
Intrinsic (Chromosomal) Mechanisms of Resistance
12/10/2014 25
4. Enzymatic Inactivation• β-Lactamases
1. A metallo-based mechanism• (class B β-lactamases) require zinc
2. A serine-based mechanism• (class A, C, and D β-lactamases)
• β-lactamases compete with PBPs for the antibiotic
• Class A on plasmid and Class C is chromosomal
• In gram-negative bacteria periplasmic space
• In gram-positive bacteria are secreted
Intrinsic (Chromosomal) Mechanisms of Resistance
12/10/2014 26
Intrinsic (Chromosomal) Mechanisms of Resistance
12/10/2014 27
Acquired Mechanisms of Resistance• Efflux• Can be acquired by horizontal gene
• Target Site Modification• Chromosomal Mutation• Enzymatic Target Site Alteration
• Acquisition of New Targets• MRSA with staphylococcal cassette chromosome mec (SCCmec)
• Enzymatic Inactivation of Antibiotics• Enzymatic modifiation of aminoglycosides results from N-acetylation, O-
phosphorylation, and O-adenylation.
12/10/2014 28
12/10/2014 29
Dissemination
• Plasmids• Conjugation • Transformation • Transduction • Transposons • Integrons
12/10/2014 30
Thank you
12/10/2014 31
12/10/2014 32
12/10/2014 33
12/10/2014 34
12/10/2014 35