International Journal of Medical Sciences And Clinical Research
66
https://theusajournals.com/index.php/ijmscr
VOLUME
Vol.05 Issue02 2025
PAGE NO.
66-72
10.37547/ijmscr/Volume05Issue02-12
Evaluation and assessment of the effects of various
antibiotics and phages on staphylococcus aureus bacteria
Iskandarova Yulduzoy.Nizamitdinovna
Independent PhD Researcher, Chirchiq State University, Uzbekistan
Fayziyev V.B.
Head of the Department, Chirchiq State University, Doctor of Biological Sciences, Professor, Uzbekistan
Adilkhonova N.A.
Head of the Bacteriological Laboratory Department, Tashkent Medical Academy, Candidate of Medical Sciences, Tashkent, Uzbekistan
Sabirjonova G. S.
Assistant Lecturer, Shahrisabz Branch of the Tashkent Chemical Technology Institute, Uzbekistan
Ubaydullaev Z.J.
Doctoral Student, Chirchiq State Pedagogical University, Uzbekistan
Received:
22 December 2024;
Accepted:
24 January 2025;
Published:
26 February 2025
Abstract:
Pyogenic cocci are among the most significant pathogens responsible for both hospital-acquired and
community-acquired infections, leading to considerable morbidity and mortality worldwide. Over the past few
decades, since the advent of antibiotics, the emergence and spread of methicillin-resistant Staphylococcus aureus
(MRSA) have become a serious healthcare challenge. In this study, patients with Staphylococcal infections were
observed in two major hospitals located in Tashkent city. Monitoring revealed a sharp increase in infection rates
over the last three years, particularly in 2021-2023, with 3,695 infections recorded at the Tashkent Medical
Academy and 14,632 cases at the Republican Scientific Center for Emergency Medical Care during the same
period. Considering this situation, the effects of various antibiotics and bacteriophages as alternative treatments
for Staphylococcus infections were studied. Results showed significant effectiveness with Cefoperazone +
Sulbactam (2.8 cm), Fosfosin (2.7 cm), Cefepime (2.7 cm), Amikacin (2.6 cm), Levofloxacin (2.7 cm), Doxycycline
(2.6 cm each), and Levomycetin (2.7 cm). The bacteriophage demonstrated an efficacy of 1.5 cm within 12 hours.
Keywords:
Staphylococcus, bacteriophages, enterocolitis, pathogenic, lysis, pyogenic, embryo, sterile, colony,
virulent.
Introduction:
Among epidemiologists, microbiologists,
and clinicians, there is a widely accepted notion that
virtually no non-pathogenic Staphylococcus species
exist [2]. However, there have been cases where
Staphylococcus isolates from blood, tissues, and organs
exhibit no pathogenic traits [4]. Interestingly, when
these bacteria are removed from the div, signs of
illness often disappear. This phenomenon should be
considered
in
the
laboratory
diagnosis
of
staphylococcal infections [5]. Unfortunately, in our
country, routine bacteriological laboratories can only
identify Staphylococcus aureus, Staphylococcus
epidermidis, and Staphylococcus saprophyticus.
Staphylococci primarily affect the skin, skin
appendages, and subcutaneous tissues [6]. They are
known to cause conditions such as furuncles,
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International Journal of Medical Sciences And Clinical Research (ISSN: 2771-2265)
carbuncles, felons, paronychia, abscesses, phlegmons,
mastitis, lymphadenitis, and wound suppuration in
operating rooms [9]. In children, staphylococci are
observed as causative agents of staphylodermia,
epidemic pustules, and impetigo. They can also cause
diseases such as pleuritis, bronchitis, pneumonia, and
peritonitis [7]. Staphylococci may lead to tonsillitis,
sinusitis, otitis, conjunctivitis, and, in rare cases,
infections such as meningitis, brain abscess,
myocarditis, endocarditis, arthritis, and infections in
vascular prostheses. Dangerous cases of foodborne
poisoning, enterocolitis, cholecystitis, cystitis, pyelitis,
and pyelonephritis have also been linked to
staphylococcal infections [11]. These microorganisms
can enter the bloodstream and bone marrow, resulting
in sepsis, osteomyelitis, and toxic shock syndrome. The
etiology of staphylococcal infections is generally not
considered to be acute infectious [10]. The growing
antibiotic resistance of microorganisms, particularly
staphylococci, has become increasingly significant in
treating these diseases [9].
Bacteriophages, or simply "phages,"
are viruses
capable of destroying bacteria, with the following
advantages:
1. Target specificity
: Phages target specific bacteria
without harming human cells or beneficial microflora,
potentially making them safer than antibiotics.
2. Effectiveness against resistance
: Bacteria may
develop resistance to antibiotics, but phages co-evolve
with bacteria, making them effective against resistant
strains.
3. Natural eliminators
: Phages naturally exist in
ecological
systems,
making
them
safe
and
environmentally friendly.
4. Fewer side effects
: Unlike antibiotics, phages only
affect targeted bacteria, leading to fewer side effects.
5. O Wide research potential
: Research on phages is
ongoing, and since their capabilities are not yet fully
explored, there is potential for developing new
treatments [13].
In the search for alternative laboratory diagnostic
methods, phage diagnostics stand out as a labor-
efficient, rapid method applicable at all laboratory
levels. The use of bacteriophages in surgery plays a
significant role in treating infected wounds, as it
directly influences the infection process. The
application of bacteriophages, discovered by Félix
d'Hérelle in 1917, opened new horizons in surgical
practices. Thousands of cases in France were treated
with bacteriophages, demonstrating the extensive
experience supporting phage therapy [11]. Based on
these scientific findings, it was decided to apply
bacteriophages
for
patients
with
identified
Staphylococcus infections. We know that the number
of antibiotics used in combating infections is increasing,
along with their adverse effects on the div.
Antibiotics tend to reduce immune system indicators,
diminishing their effectiveness against infections as
bacteria quickly adapt to them [2]. Therefore, the
primary objective of this study is to isolate
Staphylococcus bacteriophages and environmental
samples. The research was conducted in the Purulent
Surgical Department at Tashkent Medical Academy,
employing the H. G. Huxley method [12].
METHODS
At the beginning of the study, Staphylococcus was
cultivated in a nutrient medium. Bacteriological
examination of the purulent material was conducted
on days 3 and 5 before and after infection, ensuring
that the sample was introduced into the nutrient
medium within 1-2 hours of collection to prevent the
infection from deteriorating. The purulent material was
collected using sterile cotton and, after initial
microscopy, the grown colonies were transferred to a
solid nutrient medium using the Gold method for
further quantification. A swab was then immersed in
sugar broth. If there was no growth on the solid
nutrient medium, the study continued by transferring
from the sugar broth to the solid nutrient medium. The
following nutrient media were used to isolate aerobic
microorganisms: 5% blood agar, egg-yolk-salt agar, and
Sabouraud agar. Once growth was observed, the
number of colonies was calculated and recalculated to
determine the level of colonization, expressed as CFU
per swab or CFU per ml. The equipment used is
specified. Cultures on CA and JSA were incubated under
normal conditions for 18-24 hours at 1-37°C. The
progression of the infection was monitored. In blood
agar, infection growth was observed within 24 hours,
and in JSA, Staphylococcus revealed its morphology
within 48 hours.
RESULT
Using these methods, Staphylococcus infection was
examined. Based on the analysis of results,
Staphylococcus bacteria were identified. The findings
were prepared in a table format from analyses of
samples obtained from the Purulent Wound Surgery
Department of the Tashkent Medical Academy.
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International Journal of Medical Sciences And Clinical Research (ISSN: 2771-2265)
Table 1
Infections Identified in Analyses from the Purulent Wound Surgery Department
№ Name of Analysis
Type of Infection
Type of Infection
1
Throat
Staphylococeus
aureus
Candida albicans
2
Urine
+
3
Nasal cavity
+
4
Ear
+
5
Pertussis
+
6
Stool
+
7
Intestinal flora in
stool
+
8
Uppae leg wound
+
9
Gluteal wound
+
10 Lung abscess
+
11 Sputum
+
12 Thigh wound
+
13 Abdominal cavity
+
14 Index finger wound +
15 Vagina
+
16 Breast mastitis
+
17 Skin
+
18 Pus
+
19 Big toe
+
20 Cervical canal
+
21 Tonsils
+
22 Mother’s milk
+
23 Prostatic fluid
+
24 Cerebrospinal fluid
+
25 Hemoculture
+
The table presents data on patients from the purulent
wound surgery department where Staphylococcus
aureus was identified in their analyses. Specifically,
Staphylococcus aureus was detected in samples taken
from the big toe. Additionally, in women,
Staphylococcus infection was found in samples from
mastitis nodules that intensified and became purulent
during the postpartum period. Due to severe
respiratory infections, lung abscesses formed, and
analyses from this purulent material also confirmed the
presence of Staphylococcus. The site and type of
analysis are shown in the table, along with the growth
in the nutrient medium (Figure 1).
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Figure 1. Appearance of Staphylococcus aureus bacteria in Endo nutrient medium.
The numbers indicate the patients' test results.When
samples from patients in the Purulent Wound Surgery
Department of the Academy were inoculated into the
nutrient medium, bacterial growth was observed
(Figure 1). These microorganisms were cultured as a
lawn, and after growth, various antibiotic diffusion
discs were placed on the microorganisms to monitor
their activity. The results are shown below in Figure 2.
Figure 2. The effect of antibiotics on Staphylococcus infection
In this case, the numbers beneath indicate the
antibiotics that had the most effective impact.
Cefperazone+sulbactam showed a good effect with a
2.8 cm inhibition zone, Fosfomycin exhibited a 2.6 cm
effect. Cefepime also showed a 2.7 cm zone, Amikacin
2.6 cm, Levofloxacin 2.6 cm, Doxycycline 2.6 cm, and
Chloramphenicol 2.7 cm showed their efficacy. All of
these results are displayed in Figure 2. Antibiotics are
working well, showing 50-60% effectiveness.
Currently, leading researchers and scientific centers
involved in phage therapy include the Eliava Institute
(Georgia), which has been a center for Georgian phage
therapy since the 1920s and continues to conduct
extensive research on treating various bacterial
infections using phages. Texas A&M University (USA)
actively conducts research on bacteriophage therapy,
and along with other research centers, is exploring the
combined use of phages with antibiotics. Yale
University (USA), particularly in the engineering field,
has research on creating effective phages against
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International Journal of Medical Sciences And Clinical Research (ISSN: 2771-2265)
bacteria. Biotechnology companies like Phagelux,
Adaptive
Phage
Therapeutics,
and
Armata
Pharmaceuticals are developing alternative treatments
to antibiotics through phage therapy in the
pharmaceutical industry. Institut Pasteur (France) is
also renowned for its research on phages and the
development of new phage therapy strategies. These
centers and scientists play a leading role in developing
phage therapy as an effective alternative to traditional
antibiotics. Several scientific centers are conducting
research on combating antibiotic-resistant infections
using bacteriophages. For instance, the University of
California, San Diego School of Medicine (UCSD) is
attempting to use bacteriophages effectively in clinical
trials. These studies, conducted over many years for
patients with limited treatment options, help to
explore the impact of phage therapy on the immune
system, determine dosages, and durations. Research
shows that bacteriophages are not only effective
against antibiotic-resistant bacteria but also exhibit less
resistance. Additionally, the World Health Organization
(WHO) supports exploring the possibilities of
bacteriophage therapy as an alternative to antibiotics
and has launched several research programs to gather
more scientific evidence in this area. They are
specifically studying the effectiveness of using
bacteriophages to reduce antibiotic-resistant bacteria
in the human div. Also, Zhengzhou University in China
is conducting research to expand the use of
bacteriophages in combating bacteria like antibiotic-
resistant Staphylococcus aureus. These studies
propose new methods for treating highly resistant
bacteria using bacteriophages and suggest that phage
therapy could be an important tool in combating
infections resistant to antibiotics in the future, with
ongoing scientific research in this field. The properties
of bacteriophages and their significance in the
epidemiology of infectious diseases are important
areas of research. The interaction between
bacteriophages and bacteria demonstrates an effective
impact in treating diseases. The first studies in this area
were conducted by D. Herelle Felix, who studied
cultures of bacteria grown with bacteriophages on solid
nutrient media (e.g., agar-agar)[11]. During these
studies, Felix discovered that bacteria dissolve under
the influence of bacteriophages, forming sterile "spots"
that result from the dissolution of bacterial colonies
under
the
effect
of
bacteriophages.
Once
bacteriophages enter a bacterium, they begin to
produce a special enzyme
—
lysin
—
which breaks down
bacterial proteins and increases internal pressure,
causing the bacterium to burst and release a large
number of new bacteriophages. After a bacterium
bursts, between 12 to 87 new bacteriophages can be
produced. Research has shown that the reproduction
rate of bacteriophages depends on their virulence.
Highly virulent bacteriophages can double the number
of new progeny within 10-20 minutes, while less
virulent phages take 40-60 minutes. This depends on
the type and properties of the phages. Our research
tested the effectiveness of phage therapy against
infections, comparable to antibiotics. After detecting
the infection, it was inoculated in a Petri dish using the
lawn method, and Mediaphag Staphylococcus liquid
bacteriophage was applied. Within 12 hours, our
bacteriophage showed an efficacy of 1.4 cm. This
method confirms the effectiveness of phage therapy.
When observing bacteriophages under a microscope,
swelling and bursting of bacterial cells are observed 30-
35 minutes after being added to the bacterial culture.
After the cells disintegrate, the liquid contains fine
granules and amorphous masses instead of bacteria.
Once the bacteria are completely dissolved, the liquid
appears completely transparent, and ultramicroscopic
bacteriophage embryos are present. Phage therapy
plays a significant role in combating diseases such as
cholera, typhoid, and dysentery.
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International Journal of Medical Sciences And Clinical Research (ISSN: 2771-2265)
Figure 3. The effect of bacteriophage on Staphylococcus aureus.
We tested the effect of bacteriophage on the
Staphylococcus infection identified in our dish. The
impact of the bacteriophage on the infection was
observed within 12 hours, with the bacteriophage
inhibition zone reaching up to 1.4 cm. This
demonstrates the effectiveness of the bacteriophage,
yielding positive results using MediPhag's liquid
bacteriophage specific to Staphylococcus aureus.
CONCLUSION
Although antibiotics are used as an aid against infection
in the div, their side effects can weaken the immune
system and reduce the div's ability to combat
infections. Antibiotics provide only a temporary effect
without fully eliminating the infection, which may lead
to the recurrence of infection. In contrast,
bacteriophages target only the infection without
harming the div's immune system. Observations
show that the Staphylococcus aureus bacteriophage
from MediPhag demonstrated high efficacy and yielded
more positive results compared to antibiotics. Notably,
Cefoperazone + Sulbactam (2.8 cm), Fosfomycin (2.6
cm), Cefepime (2.7 cm), Amikacin (2.6 cm),
Levofloxacin (2.7 cm), Doxycycline (2.6 cm each), and
Chloramphenicol (2.7 cm) showed effectiveness. The
bacteriophage, within 12 hours, displayed its effect
with an efficiency of 1.4 cm.
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