Azithromycin

Resistance through
Interspecific Acquisition

2022
of an Epistasis-

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Dependent Efflux Pump
Component and
Transcriptional
Regulator in Neisseria
gonorrhoeae
Farah M Arif
P20211007
Introduction:

2022
Neisseria gonorrhea

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• Gram negative
• Diplococci
• Sexually transmitted bacteria
• And it is highly pathogenic
 2022
ABOUT DNA DAY
Azithromycin
Macrolide
Inhibits protein synthesis
23s rRNA of the 50S ribosome
Reason of resistance

It’s resistance is due to mutation in

the 23S rRNA at azithromycin binding
site i.e
(C2611T and A2059T).
 2022
Gonococci acquire resistance
due to
Natural competence
Intergenus recombination
mtr pump

2022
ABOUT DNA DAY
The mtr pump is composed of
mtrC, mtrD and mtrE cell envelope
protein, which together export
diverse antimicrobial agents from
the cell.
 2022
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mtrC120 substitution
A - T transition
Entire mtr region is a hotspot of
interspecie recombination which
introduce multiple rare and
divergent mosaic allell likely
from N meningitis and N
lactamica into gonococcal
population.
 2022
Mtr D gene

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Epistatic resistance
Materials
and methods:
 Material and methods:
Genome Bacterial
sequencing and culture
population 01 02 condition
genomics

Transformation
of mtr mosaic 03 04
allell
 RESUL
TS
AIMAN FATIMA P20211001
 Allelic diversity suggests increased interspecies admixture
at mtrRCDE:
.Increased in allelic diversity was
observed across mtrRCDE
. Highest diversity at mtrD

.Allele sharing between

N.gonorrhoeae and N.meningitidis
was observed at mtrR.
.Allele sharing between
N.gonorrhoeae and both
N,lactamica and N.polysaccharea at
mtr loci
.109 isolates with full length
mosaic mtrD

. 4 had mosaic mtrD genes ,99% identical to

N,meningitidis gene.
.5 had mosaic mtrD gene, 94-95% identical to
N,lactamica gene.
. Remaining had alleles, 91-92% identical to
N.meningitidis and N.lactamica.

. Isolates with full length mosaic mtrD had

azithromycin MIC of ≥ 0.25 µg/ml
. Isolates with full length mosaic mtrD and a mosaic
mtr had azithromycin MIC of ≥ 1µg/ml
 Epistasis between multiple mtr loci and within mtrD

3 donor strains Genome DNA for

GCGS0276,GCG 3 recipient donors able to
S0834, strains transform susceptible
GCGS0402 with 28BI, recipient to resistance
mosaic mtr GCGS0353, on media containing
alleles and GCGS0465. azithromycin
azithromycin
MICs ≥1µg/ml

In susceptible strains
Only common
GCGS0353 and GCGS0465
region inherited in
that became resistant to
all 28 BI
azithromycin, inherited with
transformants
mosaic mtrD and mtr alleles
was mtrRCDE
locus
RESULTS
continued...
Presented by : AREEBA KAMAL
Roll no. : P20211004
 “RESULTS“

Mosaic mtr
Epistasis
mutations

Regulatory and
Allelic diversity structural
mutations
 1

“Mosaic mtr mutations”

● Mosaic mtr mutations contribute to resistance to
diverse antimicrobial agents.
● They tested the susceptibilities of all strains and
transformant lines to the dye crystal violet,
detergents Triton X-100 and polymyxin B.
● A mosaic mtrD and a mosaic mtr promoter region
from either N. meningitidis-like or N. lactamica-
like donors and transformants with mosaic
mtrRCDE from GCGS0276 or GCGS0402 had
elevated MICs for these drugs compared to
susceptible recipient strains.
“Regulatory and structural
mutations”
● Regulatory and structural mutations epistatically
contribute to resistance.
● They evaluated expression before azithromycin
exposure and 120 min after the addition of a sub-
MIC dose of azithromycin (0.125 g/ml) to the
culture media.
● The presence of a mosaic mtr promoter resulted
in the significant upregulation of mtrC, mtrD, and
mtrE across conditions and upregulation of mtrR
in the absence of azithromycin.
“CONCLUSION”
 Results defining the role of
mosaic mtr in azithromycin
resistance affirm the importance
of other Neisseria species as an
antibiotic resistance reservoir for
N. gonorrhoeae.
 Antibiotic resistance can arise
through the interactions of multiple
loci.

 Interspecies mosaicism will be an

important consideration for future
development of sequence based
molecular resistance diagnostics.
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