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© 2022 al-Farabi Kazakh National University Int. j. biol. chem. (Online)
International Journal of Biology and Chemist48
IRSTI 31.23.25 https://doi.org/10.26577/ijbch.2022.v15.i1.05
*, E.M. Altuner
Kastamonu University, Kastamonu, Turkey
*e-mail: altinozedaa@gmail.com
(Received 2 March 2021; received in revised form 30 March 2022; accepted 13 May 2022)
AbstChemotherapeutic agents are substances that cure infectious diseases for many years. However,
problems arose due to drug resistance that developed over time and it was observed that the drugs failed to
stance that adversely
ststance mecha-
st
the cell. Thus, the microorganism becomes resistant to the drug used. If this resistance is inhibited, the drug
may work. A variety of chemical or natural inhibitors are available for inhibition. However, since they
cause toxicity problems, their clinical use is not currently available. Studies in this area are ongoing. There
are st
ststant
clinically isolated strains using EtBr dye. The strains that are Acinetobacter baumannii, Candida albicans,
Candida glabrata, Candida tropicalis, Klebsiella pneumoniae, Providencia rustigianii, Serratia odorif-
era, , Staphylococcus aureus and Streptococcus pneumoniae microorganisms. Cartwheel
1.0 mg/L, 1.5 mg/L, 2.0 mg/L and 2.5 mg/L). After the incubation, activation case was observed under UV
light. It is concluded that each st
Drug resistst
UV.
reached a very important stage. As a result, the
researches to discover new antibiotic hit compounds
of the important phenomena for antibiotic resistance.
Gram negative bacteria also have more
antibacterial resistance than gram positive bacteria
because gram negative bacteria have an outer
membrane in addition to the cell wall structure [1-3].
role in expelling substances from inside towards the
outside of the cells [4]. It can be present in all types
of living cells [5].
was the excretion of tetracycline in Escherichia coli
The Proteobacterial Antimicrobial Compound
published by Du et al. [12] and the Drug Metabolite
DMT is a large group of membrane transporters
present in eukaryotes, bacteria and archaea, and
includes exporters for a wide range of substrates such
as toxic compounds and metabolites [13-15].
antimicrobial compounds can accumulate inside the
cell, which may cause a cell death. Although there
they are not commonly used clinically due to their
toxicity [16].
positive or gram-negative bacteria or both [17]. The
mode of action are as follows.
49
Int. j. biol. chem. (Online) International Journal of Biology and Chemist
Verapamil inhibits ABC/MFS family,
Thioridazine inhibits NorA pump of the MFS family,
Carbonyl Cyanide m-Chlorophenyl Hydrazone
pump) family, Paroxetine inhibits RND/MFS family
and Reserpine inhibits RND/MFS family [10, 13, 18-
33].
The aim of this study is observing the presence
Enrichment of microorganisms and isolation
of pure colonies. In this study, totally 10 clinical
isolate MDR strains were used. 7 of the strains
Acinetobacter baumannii, Klebsiella
pneumoniae, Providencia rustigianii, Serratia
odorifera, , Staphylococcus aureus
and Streptococcus pneumoniae) and 3 are yeast
Candida albicans, Candida glabrata and Candida
tropicalis). Microorganisms were activated Luria-
for pure colony formation. Pure colonies obtained
in NA were transferred into sterile saline solution
0.5 McFarland standards.
Ethidium bromide (EtBr)-Tryptic Soy Agar (TSB)
were prepared. Microorganisms were inoculated on
the plates according the cartwheel method. Cartwheel
Cartwheel method
After incubation, TSB
agar plates containing EtBr were observed under UV
light.
Statistical analysis. R Studio, version 4.0.2 was
used to conduct a one-way analysis of variance
In order to obtain results, plates were observed
microorganism under UV light on TSB agar plates.
0.0 mg/L concentration is the negative control group.
Figure 2.
50
Int. j. biol. chem. (Online) International Journal of Biology and Chemist
Since the amount of EtBr dye at 0.5 mg/L was
Figure 3.
purple fluorescence started to appear. A low
,
, , ) was
on the second plate and the purple fluorescence of
C. glabrata and . Results presented
, , ,
were observed on the second plate. S. aureus and
was observed to be
For 2.0 mg/L of EtBr concentration; EtBr was
accumulated in the cells of all microorganisms,
S. odorifera,
S. pneumoniae, A. baumannii, P. rustigianii) in the
S. aureus, K. pneumoniae and
were observed on the second plate. The
C. albicans and C. tropicalis in the
third plate was higher than C. glabrata. Results
2.5 mg/L concentration is the highest
concentration. EtBr was accumulated in the cells
of all microorganisms. Because of the high EtBr
higher than other concentrations. Results presented
51
Int. j. biol. chem. (Online) International Journal of Biology and Chemist
According to the results it was observed that
cell changes between 1.0 and 2.0 mg/L of EtBr
for all strains used in the study. In all strains, the
In a previous study, which was conducted
by MDR E. coli
were observed. Cartwheel test with EtBr was
applied to microorganisms and similar results were
observed under UV light. In addition, verapamil,
thioridazine hydrochloride and phenyl arginine
in combination with some antibiotics for inhibiting
bacterial growth and positive results were obtained
[11].
In a study by Anbazhagan et al. MDR Salmonella
Typhimurium
activity was evaluated. They applied the EtBr
cartwheel method in the experiment. They prepared
MH agar plates with EtBr containing 0.0 mg/L, 0.5
mg/L, 1.0 mg/L, 1.5 mg/L and 2.0 mg/L. After the
incubation, the plates were examined under UV and
52
Int. j. biol. chem. (Online) International Journal of Biology and Chemist
inhibitor and some antibiotics. It was stated that 8
[36]. In the study by Martins et al. MDR E. coli,
Salmonella typhimurium, Salmonella enteritidis,
Enterococcus faecalis, Enterobacter aerogenes and
Staphylococcus aureus bacteria strains were used.
They applied the cartwheel method in TSB agars
with EtBr at concentrations between 0.0 mg/L and
[34]. Jiang et al. used the LightCycler 96 instrument
Listeria monocytogenes
they used [37].
activity was determined, inhibitor studies were
performed with antibiotic combinations. Studies
should continue to discover a non-toxic inhibitor
acts on. Thus, it is thought that it may overcome
the major problem of antibiotic resistance. In
was observed under EtBr UV between 1.0 mg/L
and 2.5 mg/L. It is clear that, researches about
they are important in bacterial resistance and
this study clearly presents that each strain could
inhibition researches. Various results can be
working with more various microorganisms.
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