Antimicrobial resistance mechanisms Flashcards
Define susceptibility
Organism is inhibited by the usually achievable concentrations of antimicrobial agent when the dosage recommended to treat at site of infection is used
Define susceptible dose dependent
Susceptibility depends on the dosage regimen used - to achieve levels that are clinically effective, there must be greater antimicrobial exposure than achieved with the dose used to determine the breakpoints
Define “intermediate” resistance
Response rates/zone size/MICs are lower than for susceptible isolates
Define resistance
Organism is not inhibited by the usually achievable concentrations of the agent with normal dosage schedules or fall into the range in which a specific known resistance mechanism is likely
Define MDR
Acquired non-susceptibility to ≥ 1 agent in ≥3 antimicrobial categories
Define XDR
Acquired non-susceptibility ≥ 1 agent in all but 2 or less antimicrobial categories
Name mechanisms of genetic variability of bacteria
Intracellular
Intercellular
What intracellular mechanisms can result in genetic variability of bacteria?
Homologous recombination
Mutations
What intercellular mechanisms can result in genetic variability in bacteria?
Bacterial
- transformation
- transduction
- transposition
- conjugation
Explain bacterial transformation
Bacterial lysis -> uptake of free segments of naked bacterial DNA from surroundings through the cell membrane -> combination of new DNA with pre-existing DNA -> degradation of unused DNA -> expression of new genes
Give examples of bacteria that often undergo transformation
Neisseria
Haemophilus influenza B
Strep pneumo
Which enzyme prevents bacterial transformation and how does it do this?
Deoxyribonucleases as they break down free DNA
Explain bacterial conjugation
Transfer of plasmids by a bridge like connection between two bacteria
Example: F+ bacteria connect with F- bacteria via the sex pills -> a single strand of plasmid DNA is transferred from F+ bacteria to F- bacteria -> 2 F+ bacteria
What is F factor?
Fertility factor - a bacterial plasmid that enables transfer of genetic material between bacteria
What are Hfr cells?
High frequency recombination cells - bacteria with a conjugative plasmid integrated into their chromosomal DNA
Explain bacterial transduction
The process of gene transfer between bacteria via bacteriophages
What is the difference between a prophage and a lytic phage?
Lytic phage - bacteria is destroyed
Prophage - bacteria integrates the phage DNA into its genome
Differentiate generalised vs specialised transduction
Generalised - any portion of the bacterial genome is transferred
Specialized - a specific portion of the bacterial genome is transferred
Explain generalised transduction
Bacteriophage attaches itself to the bacterial cell wall and injects its DNA into the bacterium -> cleavage of bacterial DNA and replication of viral DNA -> formation of new bacteriophages with phage capsids containing fragments of bacterial DNA -> lysis of bacterium -> release of new bacteriophages
Explain specialised transduction
Bacteriophage infects bacteria -> viral DNA incorporated into bacterial DNA at specific location but remains inactive -> when activated the viral DNA is replicated -> viral DNA and flanking bacterial DNA excised from the bacterial genome -> excised DNA is incorporated into new bacteriophage capsids -> lysis of bacterium -> release of new bacteriophages
Give examples of toxins that have their genes transferred via specialised transduction
Erythrogenic
Cholera
Diphtheria
Shiga
Botulinum
Explain bacterial transposition
Exchange of genetic information via transposons within the genome or between genomes of various bacteria
Define beta lactamases
Enzymes that inactivate beta lactam antibiotics by splitting the amide bond of the beta lactam ring
What gene encodes for beta lactamases?
Bla genes
How are beta lactamases classified?
Ambler classification (molecular)
Bush-Jacobi-Medeiros (functional)
What is the active site of class A, C and D?
Serine
What is the active site of class B?
Metallo-beta-lactamases (zinc)
Which enzymes are class A?
Penicillinases
- Broad spectrum
- Extended spectrum
- Carbapenemases
Which enzymes are class B?
Carbapenemases
Which enzymes are class C?
Cephalosporinases
Which enzymes are class D?
Oxacillinases
- Broad spectrum
- Extended spectrum
- Carbapenemases
Which classes do carbapenemases fall under?
A, B and D
What are the substrates of the broad spectrum penicillinases?
Benzylpenicillin
Aminopenicillins
Carboxypenicillins
Ureidopenicillins
Narrow spectrum cephalosporins
What are the substrates of the extended spectrum penicillinases?
Broad spectrum penicillinases
PLUS
Oxymino-beta-lactams
Aztreonam
What are the oxymino-beta-lactams?
Cefotaxime
Ceftazadime
Ceftriaxone
What are the substrates of the carbapenemase penicillinases?
Extended spectrum penicillinases
PLUS
Cephamycins
Carbapenems
Give examples of the genes involved in broad spectrum penicillinases
PC1
TEM-1
SHV-1
Which bacteria is associated with the PC1 gene?
Staph aureus
Which bacteria are associated with the TEM-1 and SHV-1 genes?
E.coli
Klein
Gram -ve’s
Give examples of the genes involved in extended spectrum penicillinases
TEM-derived
SHV-derived
CTX-M-derived
PER-1
VEB-1
VEB-2
GES-1
GES-2
IBC-2
Which bacteria are associated with the TEM, SHV and CTX-M derived genes?
Enterobacteriaceae
Which extended spectrum beta lactase genes are associated with pseudomonas aeruginosa?
PER-1
VEB-1
VEB-2
GES-1
GES-2
IBC-2
Give examples of the genes involved in carbapenemase penicillinases
KPC-1
KPC-2
KPC-3
NMC
IMI
SME family
Which carbapenemase genes are associated with klebs pneumonia?
KPC-1
KPC-2
KPC-3
Give examples of genes associated with the metal lo-beta-lactamases
NDM-1
IMP
VIM
GIM
SPM
SIM
Give examples of cephalosporinase enzymes
AmpC-type enzymes
What are the substrates of the cephalosporinases?
Extended spectrum
PLUS
Cephamycins
What is level of antibiotic resistance mediated by beta lactamases determined by?
- Rate of hydrolysis
- Affinity for antibiotic
- Amount of beta lactamase produced by the bacterial cell
- Susceptibility of the target protein to the antibiotic
- Rate of diffusion of the antibiotic into the bacterial cell
Is the bla gene inducible?
Yes
What is the bla gene carried on?
Small plasmids
Transposons
Where is the penicillinase enzyme excreted?
Extracellularly
How does penicillinase work
Hydrolysis of penicillin compounds into inactive penicilloic acid
What is the laboratory marker for an ESBL?
GNB that shows CTX resistance
How does TEM allow narrower spectrum enzymes to attack oxymino-beta-lactams?
Amino acid substitutions around active site of the enzyme to change configuration and allow access to oxymora-beta-lactam substrates
Amino acid substitutions via TEM at which positions produce the ESBL phenotype?
104
164
238
240
How many TEM-type enzymes are there?
> 220
What does SHV stand for?
Sulfhydryl Reagent Variable
How many SHV-type enzymes are there?
> 190
What are the most common positions for amino acid substitutions via SHV?
238
240
What does CTX-M stand for?
Cefotaxime Munich
Which beta lactamase inhibitor works more on CTX-M beta lactamases?
Tazobactam > clavulanic acid
How many CTX-M enzymes are there?
> 173
Which bacteria are oxacillinases mostly described in?
Pseudomonas aeruginosa
What does IRT stand for?
Inhibitor resistance TEM
How many inhibitor resistant beta lactamases are there?
> 19
What is the laboratory marker of AmpCs?
Resistance to cefoxitin
What are the 3 categories of AmpCs?
- Inducible resistance
- Non-inducible resistance
- Plasmid mediated resistance
What are the AmpC organisms?
ESCAPPE MAYHEM
Enterobacter
Serratia marcescens
Citrobacter freundii
Acinetobacter baumannii
Pseudomonas aeruginosa
Providencia stuartii
E.coli/albertii
Morganella morganii
Aeromonas
Yersinia enterocolitica
Hafnia alvei
Edwardsiella
Many others
Explain the mechanisms of AmpCs
Beta lactams produce cell wall degradation products -> NAG and NAM compete with UDP-n-acteyl-muramic acid peptides to bind to AmpR -> disabled AmpR -> decreased AmpC transcription regulation -> increased AmpC enzyme production
What is AmpR?
Negative regulatory protein that reduces AmpC beta lactamase expression
What is AmpD?
Protein responsible for cleavage of residues off cell wall degradation products to reduce their binding to AmpR and allow them to be recycled back into the cell wall synthesis pathway
What is AmpG?
AmpG transports oligopeptides involved in peptidoglycan recycling into the cytoplasm
In which AmpC regulatory gene do mutations most often occur?
AmpD > AmpR > AmpG
What are the potent AmpC producers?
HECK-YES
Hafnia alvei
Enterobacter cloacae
Citrobacter freundii
Klebsiella aerogenes
Yersinia enterocolitica