This study aimed to clinically and molecularly characterize carbapenemase-producing Enterobacteriaceae found in intensive care units in Guayaquil, Ecuador. Bacterial isolates were collected from clinical samples and tested for carbapenem resistance. Molecular analysis identified the carbapenemase genes present. Pulsed-field gel electrophoresis and enterobacterial repetitive intergenic consensus PCR were used to determine genetic relatedness of Klebsiella pneumoniae isolates. The results showed that KPC was the predominant carbapenemase gene. Seven clusters were identified among 108 K. pneumoniae isolates, with two clusters predominating between hospital units. This study enhances understanding of carbapenemase-producing bacteria and the dissemination of resistance genes in hospital settings.

1. Marcela Gomez Hoyos
Maria Daniela Freire
III semester
Pontificia Bolivariana University

2. ENTEROBACTERIACEAE
The enterobacteriaceae are a family of gran
negative bacterias, that are made of more than
100 species of bacillus or cocci morphology
They are part of the gut microbiota, this bacterias
doesn’t have cytochrome oxidase enzyme, some
produce toxines, and they need for them Good
growth simple compounds of carbon and nitrogen
and the right temperatura has to be between 22-
37°C
They can develope two types of fermentation
- Mixed acid fermentation
- Butanediolic fermentation
https://www.google.com/url?sa=i&url=https%3A%2F%2Fwww.youtube.com%2Fwatch%3Fv%3DZCep
MNpq7_Q&psig=AOvVaw24o-G-
o8c7Hc6E4StSZT3y&ust=1597864148422000&source=images&cd=vfe&ved=0CA0QjhxqFwoTCODZ34i
6pesCFQAAAAAdAAAAABAV

3. CARBAPENEMASE
Enzyme which belongs to b-
lactamasas´family.
-Hydrolytic perfil  b-lactamic
antimicrobials
Carbapenems
 Resistance ¿HOW?
- Cleave the b-lactam ring
the most clinically significant are
KPC, IMP, VIM, NDM and OXA-48

4. Objetive
was to clinically and molecularly characterize
carbapenemase producing-Enterobacteriaceae in
intensive care units (ICUs) in Guayaquil, Ecuador.

5. METHODS
1. Microbiological cultive
Its a method in wich the multiplication of a
microorganism like bacterias occurs, where
everything is prepare for the desired effect, they
are sown and nourished for their optimal
multiplication
The microbiological cultive are used to identify
which bacteria cause certain desease and to
wich antibiotic they are sensitive, determine
presence of a pathogens in the body
Isolates that show zones of inhibition were
considered resistant to cabapenems

6. METHODS
PFGE:
Is a technique that allows the separation of high
molecular weight DNA fragments, so that it pulls
by pulses the sequence that is being separated.
In this case, the PFGE allows the identification of
genetic variants among enterobacteria.
*The strains with similarity >= 80% were assigned to
the same group in ERIC-PCR and PFGE

7. METHODS
MIC inhibitory concentration
we use MIC to know the specific concentration of an
antibiotic that can inhibit the development or growth
of a pathogen.
We use the lower concentration to block the pathogen
growth
Its divided into sensitive, intermediate and resistant
It have been used to prove all the enteriobacteriaes
that are resistants to carbapenems
Positive MIC means that there is production of
carbapenem

8. METHODS
ERIC-PCR:
ERIC PCR  multiple bacterial species, for
which heterogeneous profiles have been
obtained to differentiate genotypically the
analyzed isolates.
The genetic relatedness between
K.pneumoniae bla KPC was determined by
ERIC PCR, in order to analyze strain from each
electrophoretic pattern obtained in ERIC PCR
with PFGE.
No other carbapenemase was studied

9. RESULTS
This table show us the
molecular mecanism of the
cabapenems resistence and the
carbapenemase detected

10. RESULTS
This figure shows the PFGE
results for all K.pneumoniae
isolates, shows detailed data
of cluster subtypes obtained
in figure 2
It can be observed that the
genetic distance between
K.pneumoniae isolates are
not so marked except for:

11. RESULTS
“PFGE identified 7 clusters among 108
isolates analysed. 2 PFGE clusters
were predominantly detected by using
a cut-off of 80% similarity in the
hospital units studied”
We can observe the similar patterns

12. RESULTS
This figure shows the variability
between one hospital and another in
terms of the number of strains and the
distribution of electrophoresis
patterns.
-Cluster A (n = 45; 17 subtypes) including
41.67% of insulation
-Cluster B (n = 40; 7 subtypes) including
37,03% of insulation
-The rest of the isolates were
distributed in cluster C (n=1;2
subtypes), D (n= 15; 6 subtypes), F
(n=1), G (n =2) and J (n = 4; 2
subtypes)

13. • DISCUSSION
da Silva R, Maciel W, Correia F,
Godoy C, Rodigues-Costa F, et al.
Risk factors for KPC-producin
Klebsiella pneumoniae
Previous studies have demonstrated an association between
KPC-producing K. pneumoniae infection and the length of
hospitalization, use of central venous catheters, ICU stay, and
exposure to specific broad-spectrum antimicrobial agents,
such as carbapenems, cephalosporins, fluoroquinolones, and
penicillin, as well as having surgery
AGREE
Papadimitriou-Olivgeris M,
Marangos M, Fligou F, et al. KPC-
producing Klebsiella pneumoniae
enteric colonization acquired
during intensive care unit stay
Debbey BD, Ganor O, Yasmin M,
et al. Epidemiology of
carbapenemn resistan Klebsiella
pneumoniae colonization in an
intensive care unit
This study showed higher mortality rates in CPE
colonized/infected patients than in CPE negative patients but
did not demonstrate differences between CPE colonized and
infected patients. These results differ from those reported by
other authors, where the length of the ICU stay was
considered an independent risk factor, but not KPC
colonization
NO
Dautzenberg MJD, Wekesa AN,
Gniadkowski M, Antoniadou A,
Giamarellou H, Petrikkos GL, et
al. The association between
colonization with
carbapenemaseproducing
Enterobacteriaceae and overall
ICU mortality
Colonization is a wellknown risk factor for a worse outcome;
a study performed in Greek intensive careunits showed
thatCPE colonized patients are at a 1.79 times more
significant risk of dying than non-colonized patients
AGREE

14. CONCLUSIONS
- The use of molecular biology to identify the alleles of KPC (in this
case), can help to understand the dissemination routes and control
the spread of this type of enzyme such as carbapenemase
Enterobacteriaceae responsible for mortality in critically ill patients.
- Studying the molecular mechanisms involved in antibiotic resistance
allows us to identify the reason for this resistance and thus have the
possibility of preventing failures in therapeutic treatment.
- The genotypic characterization by means of the different types of
electrophoresis helps to determine the genetic variability of the studied
organisms, which allows to differentiate their function and their
incidence in the patient.

15. Daniela freire
https://app.lucidchart.com/invitations/accept/cfe7e15a-0a4a-4a4e-bc37-f225b2eebfd9

16. Marcela Gomez Hoyos