Journal Reading

Characterization of Ciprofloxacin-resistant and ESBL-producing Salmonella enteric

Serotype Derby in Eastern China

Zhaojun Xu, Qifa Song, Chunhua Li and Yefei Zhan

BMC Microbiology (2019) 19:61

Sitti Monica Astrilia Ambon

Magister Program of Biomedical Science

Faculty of Medicine of Andalas University
2019
 Introduction
Treatment
 Fluoroquinolones and β-lactams
are recommended for treat
salmonellosis.
 Resistance to these drugs :
serious clinical outcomes.
Background
 Non-typhoidal Salmonella species
contaminate food  leading causes
of human foodborne diseases.
 Non-typhoidal Salmonella : serotype
Derby.
Aimed
• 1st in the present study : S. Derby were
highly resistant to fluoroquinolones and β-
lactams (ESBL-producing).
• Present study : Characterize the
mechanisms of antimicrobial resistance &
distribution patterns of the resistant and
ESBL-producing.
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 Methods

Bacterial Isolates Antimicrobial Susceptibility Testing (AST)

- Samples : Stool, food and river water. - All S. Derby isolates : Tested for susceptibility to 9
ABs (ampicilin, chloramphenicol, tetracycline,
- Stool : First put in tetrathionate brilliant-green broth or
ciprofloxacin, nalidixic acid, gentamicin, cefotaxime,
selenite broth  cultivation on Salmonella-Shigella agar
meropenem, sulfametoxazole/trimethoprim) using
/ CHROMagar Salmonella agar.
Kirby-Bauer disc diffusion method.
- Food : Buffered peptone water, selenite cystine & broth
- AST results  interpreted according to the guidelines
tetrathionate  cultivation on CHROMagar Salmonella
in the Clinical and Laboratory Standard Institute
agar.
(CLSI).
- Water : Filtered using membranes of 0.45 mcm pore
size  enrichment & cultivation same as food samples.
Food & water  environment sources.
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 Methods
Antimicrobial resistance-relevant genetic
- Three β-lactamase genes blaTEM,blaOXA &
blaCTX-M detected using PCR primers.
- Genetic mechanisms for fluoroquinolone
resistance (gyrA, qnrB & ParC genes) were
amplified and sequenced in all isolates using
primers.
Pulsed-field gel electrophoresis (PFGE)
- All S. Derby isolates were subtyped by
PFGE using the restriction enzyme XbaI
following the PulseNet standardized
protocol.
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 Methods

Plasmid Profile and Conjugation Experiment

 Fluoroquinolone-resistant and ESBL-producing S. Derby isolates were
inoculated into 200 mL of Luria-Bertani (LB) broth & cultivated for 12h with
shaking of 150 rpm.

 The plasmids in 200 mL extracted by a large-volume preparation method.

 Plasmid profiles were obtained for each isolates using electrophoresis.

 Plasmid transformation into E.coli DH5a  to assess β-lactam ressistant

dissemmination by plasmid.

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 Results
Bacterial Isolation

Patients
826 non-typhoidal Salmonella isolates
& 52 (6%) S. Derby isolate were
collected Time From 2013 to 2017

- Top 3 Salmonella serotypes : S.

Typhimurium (242 isolates, 29 %), S.
Enteritidis (95 isolates, 12 %), S. Derby
(52 isolates).
- S. Derby (30 from feces, 22 from
environmental sources) from 2200
diarrheal patients 6
 Results
AST Results

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 Results

AST Results
 52 S. Derby isolates : Only 8 isolates were susceptible to all
antimicrobials, whereas 44 isolates were resistant to at least one
antimicrobial agent (table 1).
 Two isolates (Salm1125 & Salm 1165) from feces of diarrheal patients and
one isolate (Salm 1184) from river water  identified for fluoroquinolone-
resistant and ESBL-producing.
 No significant difference for each antimicrobial agent was observed between
patient and environmental samples.
 Total resistance events in patient isolates was significantly higher than the
environmental isolates (Pearson chi square test; p=0.04).
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 Results

Analysis of Three Highly Antimicrobial-resistant S. Derby isolates

 Three S. Derby isolates (Salm1125, Salm 1165, Salm1184) were further analyzed.
 All the 3 isolates showed the same pan-resistance profile to 9 antimicrobial agents
tested  resistant to all agents.
 Genetic structures for quinolone resistance were detected : a silent gyrA mutation
S (TCC) 83S (TCT) encoding the same amino acid and three PMQR genes
qnrB, qnrS and aac(6′)-Ib-cr.
 β-lactam resistance genes : Salm1125 & Salm1165 (blaTEM, blaCTX-M, and
blaOXA), Salm1184 (blaCTX-M and blaOXA).
 β-lactam resistance was not able to spread among isolates.
 The transformation experiment was also unable to transfer plasmid
conferring ampicillin resistanceinto E. coli DH5a.
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Results

Analysis of Three Highly Antimicrobial-resistant S.

Derby isolates
• PFGE patterns of 52 S. Derby isolates with 3 highly
antibiotic resistant.
• Each isolates shows different PFGE patterns &
could be distinguished from the pan-sensitive isolate
(Salm1395).
• Salm1125 & Salm1165 (patient sources) displayed
more genetic similarity with each other (Fig1),
whereas Salm1184 (river sources) showed a greater
difference from Salm1125 & Salm1165.

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 Results

• The electrophoresis profile also demonstrated complex plasmid pattern. Plasmid profiles of
three highly antibiotic resistant S. Derby isolates (Salm1125, Salm1165, Salm1184) and two
pan-sensitive S. Derby isolate (Salm1036 and Salm1395), showing complex plasmid
composition.
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Discussion
 Discussion

The three isolates displayed different
PFGE patterns  genetically different.
The electrophoresis profile  multiple
plasmids
S. Derby was initially found to infect
animals (pigs & turkeys)  2nd most
found Salmonella serotypes from pigs
 increase food-borne disease in
human.

Fluoroquionolone (ciprofloxacin) & β-

In China, S. Derby : 3 most common
lactam are often used to treat
serotypes of non-typhoidal Salmonella
salmonellosis. Resistant to these
from patients with diarrhea.
drugs serious clinical outcomes.
 Conclusions

The present study demonstrates that highly antibiotic-resistant S. Derby with

fluoroquinolone resistance and ESBL production has been emerging.

S. Derby has become one of the most common Salmonella serotypes and
causes illness in both human & animals.

This situation gives rise to a new looming risk of food-borne diseases by S.

Derby.

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References

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Thank you