TY  - CONF
AU  - Pavić, Aleksandar
AU  - Đuriš, Jelena
AU  - Vukotić, Goran
AU  - Obradović, Mina
AU  - Plačkić, Nikola
PY  - 2024
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2379
AB  - Fungal infections, once considered a rare disease,
have become an everyday problem in modern
societies, posing major challenges to global
health. It is estimated that more than one billion
people are affected by fungal infections and 1.6
million people succumb to these diseases every
year. Of the 600 species of fungi capable of causing
infections in humans, species of the genus
Candida cause more than 85% of infections, especially
C. albicans, which has become a serious
threat to human health in immunocompromised
and immunosuppressed individuals. Unfortunately,
the current arsenal of clinical drugs relies
on only four classes of approved drugs (polyenes,
azoles, echinocandins and allylamines), which
are only partially effective, resulting in incomplete
eradication of the fungal infection. In
addition, the serious side effects, ranging from
systemic or organ-specific toxicity to poor bioavailability
and low activity, significantly hamper
the clinical use of antifungals. These problems
call for new effective and safe antifungal agents,but also for appropriate preclinical models to accurately
study potential adverse effects on the
human population and test their efficacy against
fungal infections. In this sense, zebrafish (Danio
rerio) embryos have become one of the most
powerful preclinical animal models in infection
biology and drug discovery, offering the unique
opportunity to simultaneously monitor the safety
and efficacy of the applied molecule in real
time. With the aim of providing a preclinical platform
for the identification of new safe antifungal
drugs to effectively control C. albicans infection,
we comprehensively tested the toxicity of 13
clinical antifungal drugs in the zebrafish embryo
model. The 21 toxicity endpoints, including
survival, teratogenicity, cardiotoxicity and hepatotoxicity,
were evaluated and compared with
adverse effects described in rats and humans. Of
the clinical drugs, the efficacy of fluconazole and
voriconazole was evaluated in the zebrafish - C.
albicans model of systemic and wound biofilm
infection.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health
T1  - EMPOWERING ANTIFUNGAL DRUGS DISCOVERY THROUGH THE ZEBRAFISH-INFECTIOUS DISEASES MODELLING
EP  - 140
SP  - 140
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2379
ER  - 

@conference{
author = "Pavić, Aleksandar and Đuriš, Jelena and Vukotić, Goran and Obradović, Mina and Plačkić, Nikola",
year = "2024",
abstract = "Fungal infections, once considered a rare disease,
have become an everyday problem in modern
societies, posing major challenges to global
health. It is estimated that more than one billion
people are affected by fungal infections and 1.6
million people succumb to these diseases every
year. Of the 600 species of fungi capable of causing
infections in humans, species of the genus
Candida cause more than 85% of infections, especially
C. albicans, which has become a serious
threat to human health in immunocompromised
and immunosuppressed individuals. Unfortunately,
the current arsenal of clinical drugs relies
on only four classes of approved drugs (polyenes,
azoles, echinocandins and allylamines), which
are only partially effective, resulting in incomplete
eradication of the fungal infection. In
addition, the serious side effects, ranging from
systemic or organ-specific toxicity to poor bioavailability
and low activity, significantly hamper
the clinical use of antifungals. These problems
call for new effective and safe antifungal agents,but also for appropriate preclinical models to accurately
study potential adverse effects on the
human population and test their efficacy against
fungal infections. In this sense, zebrafish (Danio
rerio) embryos have become one of the most
powerful preclinical animal models in infection
biology and drug discovery, offering the unique
opportunity to simultaneously monitor the safety
and efficacy of the applied molecule in real
time. With the aim of providing a preclinical platform
for the identification of new safe antifungal
drugs to effectively control C. albicans infection,
we comprehensively tested the toxicity of 13
clinical antifungal drugs in the zebrafish embryo
model. The 21 toxicity endpoints, including
survival, teratogenicity, cardiotoxicity and hepatotoxicity,
were evaluated and compared with
adverse effects described in rats and humans. Of
the clinical drugs, the efficacy of fluconazole and
voriconazole was evaluated in the zebrafish - C.
albicans model of systemic and wound biofilm
infection.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health",
title = "EMPOWERING ANTIFUNGAL DRUGS DISCOVERY THROUGH THE ZEBRAFISH-INFECTIOUS DISEASES MODELLING",
pages = "140-140",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2379"
}

Pavić, A., Đuriš, J., Vukotić, G., Obradović, M.,& Plačkić, N.. (2024). EMPOWERING ANTIFUNGAL DRUGS DISCOVERY THROUGH THE ZEBRAFISH-INFECTIOUS DISEASES MODELLING. in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health
Serbian Society for Microbiology., 140-140.
https://hdl.handle.net/21.15107/rcub_imagine_2379

Pavić A, Đuriš J, Vukotić G, Obradović M, Plačkić N. EMPOWERING ANTIFUNGAL DRUGS DISCOVERY THROUGH THE ZEBRAFISH-INFECTIOUS DISEASES MODELLING. in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health. 2024;:140-140.
https://hdl.handle.net/21.15107/rcub_imagine_2379 .

Pavić, Aleksandar, Đuriš, Jelena, Vukotić, Goran, Obradović, Mina, Plačkić, Nikola, "EMPOWERING ANTIFUNGAL DRUGS DISCOVERY THROUGH THE ZEBRAFISH-INFECTIOUS DISEASES MODELLING" in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health (2024):140-140,
https://hdl.handle.net/21.15107/rcub_imagine_2379 .

TY  - CONF
AU  - Plačkić, Nikola
AU  - Kljajević, Nemanja
AU  - Obradović, Mina
AU  - Kekić, Dušan
AU  - Gajić, Ina
AU  - Stanisavljević, Nemanja
AU  - Vukotić, Goran
PY  - 2024
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2383
AB  - Anti-microbial drug resistance (AMR) is one of
the global health threats caused by the misuse
of drugs typically used to treat microbial
infections in humans, animals and plants. AMR
in nosocomial infections not only significantly
hinders treatment and endangers the patients’
lives, but also elevates the costs of healthcare.
Multiple research approaches have been initiated
to combat AMR, and one promising method
is bacteriophage therapy. Bacteriophages (phages)
are viruses that naturally exploit bacteria as
their hosts for replication and can cause cell lysis,
which makes them promising candidates for
treating the infections that do not respond to
conventional antibiotic therapies. In this study,
we screened wastewater samples from four
different collectors in Belgrade urban area for
bacteriophages active against clinically isolated
strains of two biofilm-producing bacteria that
readily persist in hospital environment - Klebsiella
pneumoniae (6 strains) and Pseudomonas aeruginosa
(2 strains). Wastewaters were screened
for phage presence through phage enrichment
process, in which bacteria were grown in a mixture
of water samples and nutrient-rich broth.
Obtained cultures were screened for antimicrobial
activity against the respective host strains,
and candidates were subjected to a first-round
plaque assay to detect the phages. Finally, the
activity of all the candidates was tested against
all strains of the same species to gain the first insight
into their host range. We discovered 20 potentially
distinct bacteriophages active against
K. pneumoniae strains and two potentially different
candidates targeting P. aeruginosa. Notably,
one phage exhibited activity against all tested K.
pneumoniae strains, and four were active against
5 out of 6 tested strains. Among 22 candidates in
total, five showed depolymerizing activity, indicating
promise in combating biofilm formation.
Currently, isolation of new phages, as well as purification
and host range analysis is underway for
several candidates targeting K. pneumoniae and
two targeting P. aeruginosa strains.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health
T1  - NOVEL BACTERIOPHAGE ISOLATION FROM BELGRADE WASTEWATERS
EP  - 148
SP  - 148
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2383
ER  - 

@conference{
author = "Plačkić, Nikola and Kljajević, Nemanja and Obradović, Mina and Kekić, Dušan and Gajić, Ina and Stanisavljević, Nemanja and Vukotić, Goran",
year = "2024",
abstract = "Anti-microbial drug resistance (AMR) is one of
the global health threats caused by the misuse
of drugs typically used to treat microbial
infections in humans, animals and plants. AMR
in nosocomial infections not only significantly
hinders treatment and endangers the patients’
lives, but also elevates the costs of healthcare.
Multiple research approaches have been initiated
to combat AMR, and one promising method
is bacteriophage therapy. Bacteriophages (phages)
are viruses that naturally exploit bacteria as
their hosts for replication and can cause cell lysis,
which makes them promising candidates for
treating the infections that do not respond to
conventional antibiotic therapies. In this study,
we screened wastewater samples from four
different collectors in Belgrade urban area for
bacteriophages active against clinically isolated
strains of two biofilm-producing bacteria that
readily persist in hospital environment - Klebsiella
pneumoniae (6 strains) and Pseudomonas aeruginosa
(2 strains). Wastewaters were screened
for phage presence through phage enrichment
process, in which bacteria were grown in a mixture
of water samples and nutrient-rich broth.
Obtained cultures were screened for antimicrobial
activity against the respective host strains,
and candidates were subjected to a first-round
plaque assay to detect the phages. Finally, the
activity of all the candidates was tested against
all strains of the same species to gain the first insight
into their host range. We discovered 20 potentially
distinct bacteriophages active against
K. pneumoniae strains and two potentially different
candidates targeting P. aeruginosa. Notably,
one phage exhibited activity against all tested K.
pneumoniae strains, and four were active against
5 out of 6 tested strains. Among 22 candidates in
total, five showed depolymerizing activity, indicating
promise in combating biofilm formation.
Currently, isolation of new phages, as well as purification
and host range analysis is underway for
several candidates targeting K. pneumoniae and
two targeting P. aeruginosa strains.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health",
title = "NOVEL BACTERIOPHAGE ISOLATION FROM BELGRADE WASTEWATERS",
pages = "148-148",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2383"
}

Plačkić, N., Kljajević, N., Obradović, M., Kekić, D., Gajić, I., Stanisavljević, N.,& Vukotić, G.. (2024). NOVEL BACTERIOPHAGE ISOLATION FROM BELGRADE WASTEWATERS. in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health
Serbian Society for Microbiology., 148-148.
https://hdl.handle.net/21.15107/rcub_imagine_2383

Plačkić N, Kljajević N, Obradović M, Kekić D, Gajić I, Stanisavljević N, Vukotić G. NOVEL BACTERIOPHAGE ISOLATION FROM BELGRADE WASTEWATERS. in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health. 2024;:148-148.
https://hdl.handle.net/21.15107/rcub_imagine_2383 .

Plačkić, Nikola, Kljajević, Nemanja, Obradović, Mina, Kekić, Dušan, Gajić, Ina, Stanisavljević, Nemanja, Vukotić, Goran, "NOVEL BACTERIOPHAGE ISOLATION FROM BELGRADE WASTEWATERS" in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health (2024):148-148,
https://hdl.handle.net/21.15107/rcub_imagine_2383 .

TY  - CONF
AU  - Vukotić, Goran
AU  - Obradović, Mina
AU  - Plačkić, Nikola
AU  - Kljajević, Nemanja
AU  - Pavić, Aleksandar
AU  - Kekić, Dušan
AU  - Gajić, Ina
AU  - Kojić, Milan
AU  - Stanisavljević, Nemanja
PY  - 2024
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2375
AB  - Antimicrobial resistance (AMR) arises when
bacteria and other microbes stop responding
to medications. AMR is now recognized as one
of serious global health threats, repeatedly
appearing in the World Health Organization’s
(WHO) lists of urgent global health challenges,
including the 2024 list. It is taking a fatal toll
– nearly 5 million deaths globally per year are
associated with AMR, encompassing 1.27 million
directly attributed to AMR. The COVID-19
pandemic paved the way for aggravation of
bacterial AMR – primarily due to enhancement
in unspecific and unjustified prescription and
use of broad-spectrum antibiotics, resulting in
what is now recognized as „silent pandemic of
AMR“. Bacteriophages (phages) are natural and
specific predators of bacteria - viruses that can
infect, replicate inside and lyse arguably any
bacteria. Their therapeutic potential is being
hastily evaluated through different approaches:
in silico, in vitro, ex vivo and in vivo – in laboratory
animals as well as in human case and clinical
studies. Although the results are promising,bacteria rapidly develop resistance against
phages, which why the isolation and research
of new phages is needed. Our work is concentrated
on three bacterial species for which critical
priority by WHO has been declared – carbapenem-
resistant Acinetobacter baumannii,
Pseudomonas aeruginosa and Klebsiella pneumoniae.
Twenty distinct pathogenic strains of
A. baumannii, 6 K. pneumoniae and 6 P. aeruginosa
were used as targets for bacteriophage
isolation, and total of 14, 22 and 8 potentially
distinct phages were collected, respectively. All
strains were nosocomial isolates obtained from
various tissues, including from terminally ill patients.
Six phages were characterized in detail.
In particular, phage vB_AbaM_ISTD was applied
against A. baumannii in zebrafish embryo
model of systemic infection, and demonstrated
powerful therapeutic potential, eradicating the
infection. Interestingly, its DNA was characterized
with highly modified thymidine (amassing
1228 Da), making it the largest non-canonical
deoxynucleoside reported so far.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health
T1  - BACTERIOPHAGES OF MULTIDRUG-RESISTANT NOSOCOMIAL PATHOGENS – BELGRADE EXPERIENCE
EP  - 121
SP  - 121
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2375
ER  - 

@conference{
author = "Vukotić, Goran and Obradović, Mina and Plačkić, Nikola and Kljajević, Nemanja and Pavić, Aleksandar and Kekić, Dušan and Gajić, Ina and Kojić, Milan and Stanisavljević, Nemanja",
year = "2024",
abstract = "Antimicrobial resistance (AMR) arises when
bacteria and other microbes stop responding
to medications. AMR is now recognized as one
of serious global health threats, repeatedly
appearing in the World Health Organization’s
(WHO) lists of urgent global health challenges,
including the 2024 list. It is taking a fatal toll
– nearly 5 million deaths globally per year are
associated with AMR, encompassing 1.27 million
directly attributed to AMR. The COVID-19
pandemic paved the way for aggravation of
bacterial AMR – primarily due to enhancement
in unspecific and unjustified prescription and
use of broad-spectrum antibiotics, resulting in
what is now recognized as „silent pandemic of
AMR“. Bacteriophages (phages) are natural and
specific predators of bacteria - viruses that can
infect, replicate inside and lyse arguably any
bacteria. Their therapeutic potential is being
hastily evaluated through different approaches:
in silico, in vitro, ex vivo and in vivo – in laboratory
animals as well as in human case and clinical
studies. Although the results are promising,bacteria rapidly develop resistance against
phages, which why the isolation and research
of new phages is needed. Our work is concentrated
on three bacterial species for which critical
priority by WHO has been declared – carbapenem-
resistant Acinetobacter baumannii,
Pseudomonas aeruginosa and Klebsiella pneumoniae.
Twenty distinct pathogenic strains of
A. baumannii, 6 K. pneumoniae and 6 P. aeruginosa
were used as targets for bacteriophage
isolation, and total of 14, 22 and 8 potentially
distinct phages were collected, respectively. All
strains were nosocomial isolates obtained from
various tissues, including from terminally ill patients.
Six phages were characterized in detail.
In particular, phage vB_AbaM_ISTD was applied
against A. baumannii in zebrafish embryo
model of systemic infection, and demonstrated
powerful therapeutic potential, eradicating the
infection. Interestingly, its DNA was characterized
with highly modified thymidine (amassing
1228 Da), making it the largest non-canonical
deoxynucleoside reported so far.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health",
title = "BACTERIOPHAGES OF MULTIDRUG-RESISTANT NOSOCOMIAL PATHOGENS – BELGRADE EXPERIENCE",
pages = "121-121",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2375"
}

Vukotić, G., Obradović, M., Plačkić, N., Kljajević, N., Pavić, A., Kekić, D., Gajić, I., Kojić, M.,& Stanisavljević, N.. (2024). BACTERIOPHAGES OF MULTIDRUG-RESISTANT NOSOCOMIAL PATHOGENS – BELGRADE EXPERIENCE. in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health
Serbian Society for Microbiology., 121-121.
https://hdl.handle.net/21.15107/rcub_imagine_2375

Vukotić G, Obradović M, Plačkić N, Kljajević N, Pavić A, Kekić D, Gajić I, Kojić M, Stanisavljević N. BACTERIOPHAGES OF MULTIDRUG-RESISTANT NOSOCOMIAL PATHOGENS – BELGRADE EXPERIENCE. in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health. 2024;:121-121.
https://hdl.handle.net/21.15107/rcub_imagine_2375 .

Vukotić, Goran, Obradović, Mina, Plačkić, Nikola, Kljajević, Nemanja, Pavić, Aleksandar, Kekić, Dušan, Gajić, Ina, Kojić, Milan, Stanisavljević, Nemanja, "BACTERIOPHAGES OF MULTIDRUG-RESISTANT NOSOCOMIAL PATHOGENS – BELGRADE EXPERIENCE" in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health (2024):121-121,
https://hdl.handle.net/21.15107/rcub_imagine_2375 .

TY  - CONF
AU  - Malešević, Milka
AU  - Ćurčić, Jovana
AU  - Gardijan, Lazar
AU  - Obradović, Mina
AU  - Stanisavljević, Nemanja
AU  - Pantić, Milena
AU  - Matijašević, Danka
PY  - 2024
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2381
AB  - Pseudomonas aeruginosa has been recognized as
a priority pathogen by World Health Organization,
due to the emergence of multidrug-resistant
(MDR) strains. Thus, new treatment options
such as antivirulence strategy is urgently needed.
This strategy is based on the disruption of quorum
sensing (QS) activity of this pathogen. The
focus of this research was to explore the anti-QS
activity of four selected medicinal mushrooms
(Lentinula edodes, Cantharellus cibarius, Trametes
versicolor and Pleurotus ostreatus) extracts on
MDR clinical isolate P. aeruginosa MMA83. Another
aim was to check their cytotoxicity on Caenorhabditis
elegans AU37 (glp-4(bn2) I; sec-1(km4).
Among three types of mushroom extracts - hot
water polysaccharide extracts (WPE), hot alkali
polysaccharide extracts (APE) and methanol
extracts (Met), APE extracts downregulated all
tested QS and virulence factors genes of P. aeruginosa
MMA83. The most prominent effect was
observed for C. cibarius APE extract, lowering
expression from 2-fold (for lasI gene) to 20-fold
for lasB gene. Extracts didn’t show cytotoxic effect
on C. elegans. The efficacy of APE extracts
in lowering the expression of QS and virulence
factors genes of P. aeruginosa MMA83 indicate
that these extracts can reduce pathogenicity of
P. aeruginosa. Also, they possess one of the desirable
biotechnology features – the absence of
cytotoxicity. Anti-QS and antivirulence effect of
APE extracts on P. aeruginosa envisages these extracts
as the promising therapeutic candidates
for the development of next-generation antivirulence
agents.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health
T1  - MEDICINAL MUSHROOM EXTRACTS ATTENUATE PSEUDOMONAS AERUGINOSA QUORUM SENSING AND VIRULENCE
EP  - 145
SP  - 145
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2381
ER  - 

@conference{
author = "Malešević, Milka and Ćurčić, Jovana and Gardijan, Lazar and Obradović, Mina and Stanisavljević, Nemanja and Pantić, Milena and Matijašević, Danka",
year = "2024",
abstract = "Pseudomonas aeruginosa has been recognized as
a priority pathogen by World Health Organization,
due to the emergence of multidrug-resistant
(MDR) strains. Thus, new treatment options
such as antivirulence strategy is urgently needed.
This strategy is based on the disruption of quorum
sensing (QS) activity of this pathogen. The
focus of this research was to explore the anti-QS
activity of four selected medicinal mushrooms
(Lentinula edodes, Cantharellus cibarius, Trametes
versicolor and Pleurotus ostreatus) extracts on
MDR clinical isolate P. aeruginosa MMA83. Another
aim was to check their cytotoxicity on Caenorhabditis
elegans AU37 (glp-4(bn2) I; sec-1(km4).
Among three types of mushroom extracts - hot
water polysaccharide extracts (WPE), hot alkali
polysaccharide extracts (APE) and methanol
extracts (Met), APE extracts downregulated all
tested QS and virulence factors genes of P. aeruginosa
MMA83. The most prominent effect was
observed for C. cibarius APE extract, lowering
expression from 2-fold (for lasI gene) to 20-fold
for lasB gene. Extracts didn’t show cytotoxic effect
on C. elegans. The efficacy of APE extracts
in lowering the expression of QS and virulence
factors genes of P. aeruginosa MMA83 indicate
that these extracts can reduce pathogenicity of
P. aeruginosa. Also, they possess one of the desirable
biotechnology features – the absence of
cytotoxicity. Anti-QS and antivirulence effect of
APE extracts on P. aeruginosa envisages these extracts
as the promising therapeutic candidates
for the development of next-generation antivirulence
agents.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health",
title = "MEDICINAL MUSHROOM EXTRACTS ATTENUATE PSEUDOMONAS AERUGINOSA QUORUM SENSING AND VIRULENCE",
pages = "145-145",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2381"
}

Malešević, M., Ćurčić, J., Gardijan, L., Obradović, M., Stanisavljević, N., Pantić, M.,& Matijašević, D.. (2024). MEDICINAL MUSHROOM EXTRACTS ATTENUATE PSEUDOMONAS AERUGINOSA QUORUM SENSING AND VIRULENCE. in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health
Serbian Society for Microbiology., 145-145.
https://hdl.handle.net/21.15107/rcub_imagine_2381

Malešević M, Ćurčić J, Gardijan L, Obradović M, Stanisavljević N, Pantić M, Matijašević D. MEDICINAL MUSHROOM EXTRACTS ATTENUATE PSEUDOMONAS AERUGINOSA QUORUM SENSING AND VIRULENCE. in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health. 2024;:145-145.
https://hdl.handle.net/21.15107/rcub_imagine_2381 .

Malešević, Milka, Ćurčić, Jovana, Gardijan, Lazar, Obradović, Mina, Stanisavljević, Nemanja, Pantić, Milena, Matijašević, Danka, "MEDICINAL MUSHROOM EXTRACTS ATTENUATE PSEUDOMONAS AERUGINOSA QUORUM SENSING AND VIRULENCE" in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health (2024):145-145,
https://hdl.handle.net/21.15107/rcub_imagine_2381 .

TY  - CONF
AU  - Malešević, Milka
AU  - Stanisavljević, Nemanja
AU  - Rašić, Slađan
AU  - Vukotić, Goran
AU  - Gardijan, Lazar
AU  - Obradović, Mina
AU  - Kojić, Milan
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2150
AB  - Introduction: Brevibacillus laterosporus is a promising microbiological agent that can be used to prevent and control destructive diseases affecting honey bee colonies. In the presentstudy, the short-term
effect of the B. laterosporus BGSP11 bee diet on microbiota and mycobiota was investigated.
Methods: The honey bee diet was supplemented with spores of B. laterosporus BGSP11 at a concentration of 1×108 CFU/mL in sucrose solution. Metabarcoding analysis of the bee microbial community profile was performed based on 16S RNA (bacteriobiota) and Internally Transcribes Spacer (ITS) region
(mycobiota) obtained using MiSeq Illumina sequencing. The QIIME2 v2021.4 pipeline was used to analyze the obtained amplicon data library.
Results: The results show that the BGSP11 bee diet slightly altered the bee microbiota and did not lead
to potentially harmful changes in the bacterial microbiota. Moreover, it can potentially induce positive
changes, mainly reflected in the reduction of opportunistic bacteria. On the other hand, the treatment
had a greater effect on mycobiota. However, the changesin the bee mycobiome caused by the treatment
cannot be considered a priori as beneficial or harmful,since the interaction between the bee and its mycobiome is not sufficiently studied. The observed positive changes in the bee mycobiome are mainly
reflected in the reduction of phytopathogenic fungi that may affect the organoleptic and techno-functional properties of honey.
Conclusion: This pilot study suggests that the introduction of BGSP11 in beekeeping practice as a biological agent could be considered due to no harmful effects observed on the microbiota of bees.
PB  - Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
C3  - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
T1  - Short-term effect of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome
EP  - 112
SP  - 112
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2150
ER  - 

@conference{
author = "Malešević, Milka and Stanisavljević, Nemanja and Rašić, Slađan and Vukotić, Goran and Gardijan, Lazar and Obradović, Mina and Kojić, Milan",
year = "2023",
abstract = "Introduction: Brevibacillus laterosporus is a promising microbiological agent that can be used to prevent and control destructive diseases affecting honey bee colonies. In the presentstudy, the short-term
effect of the B. laterosporus BGSP11 bee diet on microbiota and mycobiota was investigated.
Methods: The honey bee diet was supplemented with spores of B. laterosporus BGSP11 at a concentration of 1×108 CFU/mL in sucrose solution. Metabarcoding analysis of the bee microbial community profile was performed based on 16S RNA (bacteriobiota) and Internally Transcribes Spacer (ITS) region
(mycobiota) obtained using MiSeq Illumina sequencing. The QIIME2 v2021.4 pipeline was used to analyze the obtained amplicon data library.
Results: The results show that the BGSP11 bee diet slightly altered the bee microbiota and did not lead
to potentially harmful changes in the bacterial microbiota. Moreover, it can potentially induce positive
changes, mainly reflected in the reduction of opportunistic bacteria. On the other hand, the treatment
had a greater effect on mycobiota. However, the changesin the bee mycobiome caused by the treatment
cannot be considered a priori as beneficial or harmful,since the interaction between the bee and its mycobiome is not sufficiently studied. The observed positive changes in the bee mycobiome are mainly
reflected in the reduction of phytopathogenic fungi that may affect the organoleptic and techno-functional properties of honey.
Conclusion: This pilot study suggests that the introduction of BGSP11 in beekeeping practice as a biological agent could be considered due to no harmful effects observed on the microbiota of bees.",
publisher = "Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade",
journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia",
title = "Short-term effect of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome",
pages = "112-112",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2150"
}

Malešević, M., Stanisavljević, N., Rašić, S., Vukotić, G., Gardijan, L., Obradović, M.,& Kojić, M.. (2023). Short-term effect of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 112-112.
https://hdl.handle.net/21.15107/rcub_imagine_2150

Malešević M, Stanisavljević N, Rašić S, Vukotić G, Gardijan L, Obradović M, Kojić M. Short-term effect of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia. 2023;:112-112.
https://hdl.handle.net/21.15107/rcub_imagine_2150 .

Malešević, Milka, Stanisavljević, Nemanja, Rašić, Slađan, Vukotić, Goran, Gardijan, Lazar, Obradović, Mina, Kojić, Milan, "Short-term effect of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia (2023):112-112,
https://hdl.handle.net/21.15107/rcub_imagine_2150 .

TY  - CONF
AU  - Obradović, Mina
AU  - Pavić, Aleksandra
AU  - Vukotić, Goran
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2137
AB  - Introduction: There is an urgent demand for the development of new therapeutic approachesto combat multidrug-resistant Acinetobacter baumannii, and bacteriophages appear to be a highly promising
solution. Phages are suitable to precisely target the infection-causing bacteria without disrupting the
beneficial microbiota. The zebrafish (Danio rerio) embryo model represents an insightful animal model
for preclinical studying of various infectious diseases and for discovery of novel safe and effective antimicrobial drugs.
Methods: Systemic bacterial infection was established by microinjection of 2000 cells of nosocomial
carbapenem-resistant A. baumannii strain 6077/12 into the bloodstream of 48 hour old zebrafish embryos. Infected embryos were treated by parenteral administration of 4 different doses (10, 50, 100, 500
PFU) of bacteriophage vB_AbaM_ISTD at 6 hours after infection (hpi). Efficacy of treatment was evaluated according to embryo survival, morphological malformations and bacterial burden (CFU) over a 3-
day period.
Results: A. baumannii-infected embryos treated with bacteriophage resulted with 100% survival rate,
while 70% of untreated embryos survived to 24 hpi and none to the end of the experiment. Viable bacterial cell count and embryo morphology observations indicated that the administered phage effectively reduced A. baumanii infection in vivo. The most effective dose was 500 PFU, decreasing the bacterial
load by 3.09 log units during 24 hpi, while lower bacteriophage doses(10, 50 and 100 PFU) produced less
prominent, but also significant bacterial reduction of 2.10, 2.19 and 2.67 log units, respectively.
Conclusion: Parentheral administration of phage ISTD demonstrated potent therapeutic activity against
A. baumannii infection in every investigated dose.
PB  - Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
C3  - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
T1  - Unveiling therapeutic potential of bacteriophage treatment in acinetobacter baumannii-infected zebrafish embryo model
EP  - 83
SP  - 83
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2137
ER  - 

@conference{
author = "Obradović, Mina and Pavić, Aleksandra and Vukotić, Goran",
year = "2023",
abstract = "Introduction: There is an urgent demand for the development of new therapeutic approachesto combat multidrug-resistant Acinetobacter baumannii, and bacteriophages appear to be a highly promising
solution. Phages are suitable to precisely target the infection-causing bacteria without disrupting the
beneficial microbiota. The zebrafish (Danio rerio) embryo model represents an insightful animal model
for preclinical studying of various infectious diseases and for discovery of novel safe and effective antimicrobial drugs.
Methods: Systemic bacterial infection was established by microinjection of 2000 cells of nosocomial
carbapenem-resistant A. baumannii strain 6077/12 into the bloodstream of 48 hour old zebrafish embryos. Infected embryos were treated by parenteral administration of 4 different doses (10, 50, 100, 500
PFU) of bacteriophage vB_AbaM_ISTD at 6 hours after infection (hpi). Efficacy of treatment was evaluated according to embryo survival, morphological malformations and bacterial burden (CFU) over a 3-
day period.
Results: A. baumannii-infected embryos treated with bacteriophage resulted with 100% survival rate,
while 70% of untreated embryos survived to 24 hpi and none to the end of the experiment. Viable bacterial cell count and embryo morphology observations indicated that the administered phage effectively reduced A. baumanii infection in vivo. The most effective dose was 500 PFU, decreasing the bacterial
load by 3.09 log units during 24 hpi, while lower bacteriophage doses(10, 50 and 100 PFU) produced less
prominent, but also significant bacterial reduction of 2.10, 2.19 and 2.67 log units, respectively.
Conclusion: Parentheral administration of phage ISTD demonstrated potent therapeutic activity against
A. baumannii infection in every investigated dose.",
publisher = "Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade",
journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia",
title = "Unveiling therapeutic potential of bacteriophage treatment in acinetobacter baumannii-infected zebrafish embryo model",
pages = "83-83",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2137"
}

Obradović, M., Pavić, A.,& Vukotić, G.. (2023). Unveiling therapeutic potential of bacteriophage treatment in acinetobacter baumannii-infected zebrafish embryo model. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 83-83.
https://hdl.handle.net/21.15107/rcub_imagine_2137

Obradović M, Pavić A, Vukotić G. Unveiling therapeutic potential of bacteriophage treatment in acinetobacter baumannii-infected zebrafish embryo model. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia. 2023;:83-83.
https://hdl.handle.net/21.15107/rcub_imagine_2137 .

Obradović, Mina, Pavić, Aleksandra, Vukotić, Goran, "Unveiling therapeutic potential of bacteriophage treatment in acinetobacter baumannii-infected zebrafish embryo model" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia (2023):83-83,
https://hdl.handle.net/21.15107/rcub_imagine_2137 .

TY  - JOUR
AU  - Obradović, Mina
AU  - Malešević, Milka
AU  - Di Luca, Mariagrazia
AU  - Kekić, Dušan
AU  - Gajić, Ina
AU  - McAuliffe, Olivia
AU  - Neve, Horst
AU  - Stanisavljević, Nemanja
AU  - Vukotić, Goran
AU  - Kojić, Milan
PY  - 2023
UR  - https://www.mdpi.com/1999-4915/15/3/628
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2065
AB  - Klebsiella pneumoniae is a global health threat and bacteriophages are a potential solution in combating pandrug-resistant K. pneumoniae infections. Two lytic phages, LASTA and SJM3, active against several pandrug-resistant, nosocomial strains of K. pneumoniae were isolated and characterized. Their host range is narrow and latent period is particularly long; however, their lysogenic nature was refuted using both bioinformatic and experimental approaches. Genome sequence analysis clustered them with only two other phages into the new genus Lastavirus. Genomes of LASTA and SJM3 differ in only 13 base pairs, mainly located in tail fiber genes. Individual phages, as well as their cocktail, demonstrated significant bacterial reduction capacity in a time-dependent manner, yielding up to 4 log reduction against planktonic, and up to 2.59 log on biofilm-embedded, cells. Bacteria emerging from the contact with the phages developed resistance and achieved numbers comparable to the growth control after 24 h. The resistance to the phage seems to be of a transient nature and varies significantly between the two phages, as resistance to LASTA remained constant while resensitization to SJM3 was more prominent. Albeit with very few differences, SJM3 performed better than LASTA overall; however, more investigation is needed in order to consider them for therapeutic application.
T2  - Viruses
T1  - Isolation, Characterization, Genome Analysis and Host Resistance Development of Two Novel Lastavirus Phages Active against Pandrug-Resistant Klebsiella pneumoniae
IS  - 3
SP  - 628
VL  - 15
DO  - 10.3390/v15030628
ER  - 

@article{
author = "Obradović, Mina and Malešević, Milka and Di Luca, Mariagrazia and Kekić, Dušan and Gajić, Ina and McAuliffe, Olivia and Neve, Horst and Stanisavljević, Nemanja and Vukotić, Goran and Kojić, Milan",
year = "2023",
abstract = "Klebsiella pneumoniae is a global health threat and bacteriophages are a potential solution in combating pandrug-resistant K. pneumoniae infections. Two lytic phages, LASTA and SJM3, active against several pandrug-resistant, nosocomial strains of K. pneumoniae were isolated and characterized. Their host range is narrow and latent period is particularly long; however, their lysogenic nature was refuted using both bioinformatic and experimental approaches. Genome sequence analysis clustered them with only two other phages into the new genus Lastavirus. Genomes of LASTA and SJM3 differ in only 13 base pairs, mainly located in tail fiber genes. Individual phages, as well as their cocktail, demonstrated significant bacterial reduction capacity in a time-dependent manner, yielding up to 4 log reduction against planktonic, and up to 2.59 log on biofilm-embedded, cells. Bacteria emerging from the contact with the phages developed resistance and achieved numbers comparable to the growth control after 24 h. The resistance to the phage seems to be of a transient nature and varies significantly between the two phages, as resistance to LASTA remained constant while resensitization to SJM3 was more prominent. Albeit with very few differences, SJM3 performed better than LASTA overall; however, more investigation is needed in order to consider them for therapeutic application.",
journal = "Viruses",
title = "Isolation, Characterization, Genome Analysis and Host Resistance Development of Two Novel Lastavirus Phages Active against Pandrug-Resistant Klebsiella pneumoniae",
number = "3",
pages = "628",
volume = "15",
doi = "10.3390/v15030628"
}

Obradović, M., Malešević, M., Di Luca, M., Kekić, D., Gajić, I., McAuliffe, O., Neve, H., Stanisavljević, N., Vukotić, G.,& Kojić, M.. (2023). Isolation, Characterization, Genome Analysis and Host Resistance Development of Two Novel Lastavirus Phages Active against Pandrug-Resistant Klebsiella pneumoniae. in Viruses, 15(3), 628.
https://doi.org/10.3390/v15030628

Obradović M, Malešević M, Di Luca M, Kekić D, Gajić I, McAuliffe O, Neve H, Stanisavljević N, Vukotić G, Kojić M. Isolation, Characterization, Genome Analysis and Host Resistance Development of Two Novel Lastavirus Phages Active against Pandrug-Resistant Klebsiella pneumoniae. in Viruses. 2023;15(3):628.
doi:10.3390/v15030628 .

Obradović, Mina, Malešević, Milka, Di Luca, Mariagrazia, Kekić, Dušan, Gajić, Ina, McAuliffe, Olivia, Neve, Horst, Stanisavljević, Nemanja, Vukotić, Goran, Kojić, Milan, "Isolation, Characterization, Genome Analysis and Host Resistance Development of Two Novel Lastavirus Phages Active against Pandrug-Resistant Klebsiella pneumoniae" in Viruses, 15, no. 3 (2023):628,
https://doi.org/10.3390/v15030628 . .

TY  - DATA
AU  - Obradović, Mina
AU  - Malešević, Milka
AU  - Di Luca, Mariagrazia
AU  - Kekić, Dušan
AU  - Gajić, Ina
AU  - McAuliffe, Olivia
AU  - Neve, Horst
AU  - Stanisavljević, Nemanja
AU  - Vukotić, Goran
AU  - Kojić, Milan
PY  - 2023
UR  - https://www.mdpi.com/1999-4915/15/3/628
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2065
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2072
T2  - Viruses
T1  - Supplementary data for the article: Obradović, M., Malešević, M., Di Luca, M., Kekić, D., Gajić, I., McAuliffe, O., Neve, H., Stanisavljević, N., Vukotić, G.,& Kojić, M.. (2023). Isolation, Characterization, Genome Analysis and Host Resistance Development of Two Novel Lastavirus Phages Active against Pandrug-Resistant Klebsiella pneumoniae. in Viruses, 15(3), 628. https://doi.org/10.3390/v15030628
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2072
ER  - 

@misc{
author = "Obradović, Mina and Malešević, Milka and Di Luca, Mariagrazia and Kekić, Dušan and Gajić, Ina and McAuliffe, Olivia and Neve, Horst and Stanisavljević, Nemanja and Vukotić, Goran and Kojić, Milan",
year = "2023",
journal = "Viruses",
title = "Supplementary data for the article: Obradović, M., Malešević, M., Di Luca, M., Kekić, D., Gajić, I., McAuliffe, O., Neve, H., Stanisavljević, N., Vukotić, G.,& Kojić, M.. (2023). Isolation, Characterization, Genome Analysis and Host Resistance Development of Two Novel Lastavirus Phages Active against Pandrug-Resistant Klebsiella pneumoniae. in Viruses, 15(3), 628. https://doi.org/10.3390/v15030628",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2072"
}

Obradović, M., Malešević, M., Di Luca, M., Kekić, D., Gajić, I., McAuliffe, O., Neve, H., Stanisavljević, N., Vukotić, G.,& Kojić, M.. (2023). Supplementary data for the article: Obradović, M., Malešević, M., Di Luca, M., Kekić, D., Gajić, I., McAuliffe, O., Neve, H., Stanisavljević, N., Vukotić, G.,& Kojić, M.. (2023). Isolation, Characterization, Genome Analysis and Host Resistance Development of Two Novel Lastavirus Phages Active against Pandrug-Resistant Klebsiella pneumoniae. in Viruses, 15(3), 628. https://doi.org/10.3390/v15030628. in Viruses.
https://hdl.handle.net/21.15107/rcub_imagine_2072

Obradović M, Malešević M, Di Luca M, Kekić D, Gajić I, McAuliffe O, Neve H, Stanisavljević N, Vukotić G, Kojić M. Supplementary data for the article: Obradović, M., Malešević, M., Di Luca, M., Kekić, D., Gajić, I., McAuliffe, O., Neve, H., Stanisavljević, N., Vukotić, G.,& Kojić, M.. (2023). Isolation, Characterization, Genome Analysis and Host Resistance Development of Two Novel Lastavirus Phages Active against Pandrug-Resistant Klebsiella pneumoniae. in Viruses, 15(3), 628. https://doi.org/10.3390/v15030628. in Viruses. 2023;.
https://hdl.handle.net/21.15107/rcub_imagine_2072 .

Obradović, Mina, Malešević, Milka, Di Luca, Mariagrazia, Kekić, Dušan, Gajić, Ina, McAuliffe, Olivia, Neve, Horst, Stanisavljević, Nemanja, Vukotić, Goran, Kojić, Milan, "Supplementary data for the article: Obradović, M., Malešević, M., Di Luca, M., Kekić, D., Gajić, I., McAuliffe, O., Neve, H., Stanisavljević, N., Vukotić, G.,& Kojić, M.. (2023). Isolation, Characterization, Genome Analysis and Host Resistance Development of Two Novel Lastavirus Phages Active against Pandrug-Resistant Klebsiella pneumoniae. in Viruses, 15(3), 628. https://doi.org/10.3390/v15030628" in Viruses (2023),
https://hdl.handle.net/21.15107/rcub_imagine_2072 .

TY  - JOUR
AU  - Stanisavljević, Nemanja
AU  - Malešević, Milka
AU  - Rašić, Slađan
AU  - Vukotić, Goran
AU  - Gardijan, Lazar
AU  - Obradović, Mina
AU  - Kojić, Milan
PY  - 2023
UR  - https://doi.org/10.1080/00218839.2023.2244710
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2063
AB  - In the current study, honey bees’ diet was supplemented with spores of Brevibacillus laterosporus BGSP11 at concentration of 108 CFU/ml in sucrose solution and its short-term effects on their micro- and mycobiota have been analyzed using Illumina MiSeq sequencing. Obtained results indicate that this treatment does not lead to potentially harmful changes in the bacterial microbiome of worker bees, slightly affecting the composition of core microbiota. Moreover, several potentially beneficial changes have been observed. The treatment has led to a significant increase in the abundance of Snodgrassella alvi, and species from Lactobacillus and Bifidobacterium genera which play important roles in protection against several honey bee pathogens. Simultaneously, B. laterosporus enriched diet have led to almost complete eradication of Enterobacteriaceae family, the taxon that contains several putative pathogen species. On the other hand, the treatment affected mycobiota more profoundly, which was expected considering the greater instability compared to microbiota. Although the observed changes in honey bee mycobiome cannot be considered a priori beneficial or harmful, since the interaction between the bee and its mycobiome has not been sufficiently studied, certain beneficial consequences of the treatment have been observed. They are primarily reflected in the reduction of phytopathogenic fungi that can affect the organoleptic and techno-functional characteristics of honey. In addition, before introducing B. laterosporus in beekeeping practice as a biological agent for pathogen control it is necessary to perform more thorough studies of the impact on the honey bee microbiome, immune system, physiology and economic characteristics of honey bee colonies.
T2  - Journal of Apicultural Research
T1  - Short-term effects of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome: a pilot study
DO  - 10.1080/00218839.2023.2244710
ER  - 

@article{
author = "Stanisavljević, Nemanja and Malešević, Milka and Rašić, Slađan and Vukotić, Goran and Gardijan, Lazar and Obradović, Mina and Kojić, Milan",
year = "2023",
abstract = "In the current study, honey bees’ diet was supplemented with spores of Brevibacillus laterosporus BGSP11 at concentration of 108 CFU/ml in sucrose solution and its short-term effects on their micro- and mycobiota have been analyzed using Illumina MiSeq sequencing. Obtained results indicate that this treatment does not lead to potentially harmful changes in the bacterial microbiome of worker bees, slightly affecting the composition of core microbiota. Moreover, several potentially beneficial changes have been observed. The treatment has led to a significant increase in the abundance of Snodgrassella alvi, and species from Lactobacillus and Bifidobacterium genera which play important roles in protection against several honey bee pathogens. Simultaneously, B. laterosporus enriched diet have led to almost complete eradication of Enterobacteriaceae family, the taxon that contains several putative pathogen species. On the other hand, the treatment affected mycobiota more profoundly, which was expected considering the greater instability compared to microbiota. Although the observed changes in honey bee mycobiome cannot be considered a priori beneficial or harmful, since the interaction between the bee and its mycobiome has not been sufficiently studied, certain beneficial consequences of the treatment have been observed. They are primarily reflected in the reduction of phytopathogenic fungi that can affect the organoleptic and techno-functional characteristics of honey. In addition, before introducing B. laterosporus in beekeeping practice as a biological agent for pathogen control it is necessary to perform more thorough studies of the impact on the honey bee microbiome, immune system, physiology and economic characteristics of honey bee colonies.",
journal = "Journal of Apicultural Research",
title = "Short-term effects of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome: a pilot study",
doi = "10.1080/00218839.2023.2244710"
}

Stanisavljević, N., Malešević, M., Rašić, S., Vukotić, G., Gardijan, L., Obradović, M.,& Kojić, M.. (2023). Short-term effects of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome: a pilot study. in Journal of Apicultural Research.
https://doi.org/10.1080/00218839.2023.2244710

Stanisavljević N, Malešević M, Rašić S, Vukotić G, Gardijan L, Obradović M, Kojić M. Short-term effects of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome: a pilot study. in Journal of Apicultural Research. 2023;.
doi:10.1080/00218839.2023.2244710 .

Stanisavljević, Nemanja, Malešević, Milka, Rašić, Slađan, Vukotić, Goran, Gardijan, Lazar, Obradović, Mina, Kojić, Milan, "Short-term effects of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome: a pilot study" in Journal of Apicultural Research (2023),
https://doi.org/10.1080/00218839.2023.2244710 . .

TY  - JOUR
AU  - Gasić, Katarina
AU  - Obradović, Mina
AU  - Kuzmanović, Nemanja
AU  - Zlatković, Nevena
AU  - Ivanović, Milan
AU  - Ristić, Danijela
AU  - Obradović, Aleksa
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1571
AB  - Bacterial fruit blotch and seedling blight, caused by Acidovorax citrulli, is one of the most destructive diseases of melon and watermelon in many countries. Pathogen-free seed and cultural practices are major pillars of the disease control. However, use of bacteriophages as natural biocontrol agents might also contribute to the disease management. Therefore, we isolated 12 bacteriophages specific to A. citrulli, from phyllosphere and rhizosphere of diseased watermelon plants. The phage strains were characterized based on their host range, plaque and virion morphology, thermal inactivation point, adsorption rate, one step growth curve, restriction fragment length polymorphism (RFLP), and genomic analysis. Transmission electron microscopy of three phage strains indicated that they belong to the order Caudovirales, family Siphoviridae. All phages lysed 30 out of 32 tested A. citrulli strains isolated in Serbia, and did not lyse other less related bacterial species. They produced clear plaques, 2 mm in diameter, on bacterial lawns of different A. citrulli strains after 24 h of incubation. The thermal inactivation point was 66 or 67 degrees C. They were stable at pH 5-9, but were sensitive to chloroform and inactivated in either 5 or 10 min exposure to ultraviolet (UV) light. RFLP analysis using EcoRI, BsmI and BamHI enzymes did not show genetic differences among the tested phages. Adsorption rate and one step growth curve were determined for the Acidovorax phage ACF1. Draft genome sequence of the ACF1 phage was 59.377 bp in size, with guanine-cytosine (GC) content 64.5%, including 89 open reading frames. This phage shared a very high genomic identity with Acidovorax phage ACPWH, isolated in South Korea. Evaluation of systemic nature of ACF1 strain showed that it can be absorbed by roots and translocated to upper parts of watermelon plants where it survived up to 10 days.
PB  - Frontiers Media Sa, Lausanne
T2  - Frontiers in Microbiology
T1  - Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli
VL  - 12
DO  - 10.3389/fmicb.2021.803789
ER  - 

@article{
author = "Gasić, Katarina and Obradović, Mina and Kuzmanović, Nemanja and Zlatković, Nevena and Ivanović, Milan and Ristić, Danijela and Obradović, Aleksa",
year = "2022",
abstract = "Bacterial fruit blotch and seedling blight, caused by Acidovorax citrulli, is one of the most destructive diseases of melon and watermelon in many countries. Pathogen-free seed and cultural practices are major pillars of the disease control. However, use of bacteriophages as natural biocontrol agents might also contribute to the disease management. Therefore, we isolated 12 bacteriophages specific to A. citrulli, from phyllosphere and rhizosphere of diseased watermelon plants. The phage strains were characterized based on their host range, plaque and virion morphology, thermal inactivation point, adsorption rate, one step growth curve, restriction fragment length polymorphism (RFLP), and genomic analysis. Transmission electron microscopy of three phage strains indicated that they belong to the order Caudovirales, family Siphoviridae. All phages lysed 30 out of 32 tested A. citrulli strains isolated in Serbia, and did not lyse other less related bacterial species. They produced clear plaques, 2 mm in diameter, on bacterial lawns of different A. citrulli strains after 24 h of incubation. The thermal inactivation point was 66 or 67 degrees C. They were stable at pH 5-9, but were sensitive to chloroform and inactivated in either 5 or 10 min exposure to ultraviolet (UV) light. RFLP analysis using EcoRI, BsmI and BamHI enzymes did not show genetic differences among the tested phages. Adsorption rate and one step growth curve were determined for the Acidovorax phage ACF1. Draft genome sequence of the ACF1 phage was 59.377 bp in size, with guanine-cytosine (GC) content 64.5%, including 89 open reading frames. This phage shared a very high genomic identity with Acidovorax phage ACPWH, isolated in South Korea. Evaluation of systemic nature of ACF1 strain showed that it can be absorbed by roots and translocated to upper parts of watermelon plants where it survived up to 10 days.",
publisher = "Frontiers Media Sa, Lausanne",
journal = "Frontiers in Microbiology",
title = "Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli",
volume = "12",
doi = "10.3389/fmicb.2021.803789"
}

Gasić, K., Obradović, M., Kuzmanović, N., Zlatković, N., Ivanović, M., Ristić, D.,& Obradović, A.. (2022). Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli. in Frontiers in Microbiology
Frontiers Media Sa, Lausanne., 12.
https://doi.org/10.3389/fmicb.2021.803789

Gasić K, Obradović M, Kuzmanović N, Zlatković N, Ivanović M, Ristić D, Obradović A. Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli. in Frontiers in Microbiology. 2022;12.
doi:10.3389/fmicb.2021.803789 .

Gasić, Katarina, Obradović, Mina, Kuzmanović, Nemanja, Zlatković, Nevena, Ivanović, Milan, Ristić, Danijela, Obradović, Aleksa, "Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli" in Frontiers in Microbiology, 12 (2022),
https://doi.org/10.3389/fmicb.2021.803789 . .

TY  - JOUR
AU  - Gardijan, Lazar
AU  - Miljković, Marija
AU  - Obradović, Mina
AU  - Borović, Branka
AU  - Vukotić, Goran
AU  - Jovanović, Goran
AU  - Kojić, Milan
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1603
AB  - Aims The aim of this study was to construct the improved pMAL expression vector to increase the efficacy of purification of small native peptides and their clear-cut separation from MBP tag. The modifications we introduced can be applied to many expression vectors. Methods and Results To improve the pMAL expression vector, we introduced the His(6) tag and the enterokinase cleavage site (Ek) downstream from the MBP tag and Xa cleavage site on the original vector. For cloning of a desired peptide DNA, the enterokinase site contains a unique BsaBI restriction site adjacent to the original multi-cloning site. This redesigned pMAL vector was optimized for the purification of cytoplasmic (pMALc5HisEk) and periplasmic (pMALp5HisEk) peptides. The purification of native and active peptide (P) was obtained following two-step affinity chromatography. In the first step, the entire MBP-His(6)-Ek-P fusion protein is purified using the Ni-NTA agarose column. This fusion protein was cleaved with active His(6) tagged enterokinase. In the second step, the further purification was performed by column containing the mixture of amylose and Ni-NTA agarose resins. This removes both the MBP-His(6) and His(6)-enterokinase leaving pure native protein in solution. These new vectors and the two-step purification protocol were successfully applied in purification of active native small antimicrobial peptides (AMPs), lactococcin A and human beta-defensin. Conclusions We constructed the improved pMAL expression vectors and established the pipeline and optimal conditions for their use in efficient purification of large amounts of active native small peptides. Significance and Impact of the Study Choice of expression vector impacts on the efficiency of expression and purification of desired proteins. The idea of redesigning pMAL vector was driven by the need for rapid purification of larger amounts of active native AMPs. This newly improved pMAL vector, the cloning strategy, expression conditions and two-step purification protocol represent a unique simple approach which can be applied in every laboratory.
PB  - Wiley, Hoboken
T2  - Journal of Applied Microbiology
T1  - Redesigned pMAL expression vector for easy and fast purification of active native antimicrobial peptides
EP  - 1013
IS  - 2
SP  - 1001
VL  - 133
DO  - 10.1111/jam.15623
ER  - 

@article{
author = "Gardijan, Lazar and Miljković, Marija and Obradović, Mina and Borović, Branka and Vukotić, Goran and Jovanović, Goran and Kojić, Milan",
year = "2022",
abstract = "Aims The aim of this study was to construct the improved pMAL expression vector to increase the efficacy of purification of small native peptides and their clear-cut separation from MBP tag. The modifications we introduced can be applied to many expression vectors. Methods and Results To improve the pMAL expression vector, we introduced the His(6) tag and the enterokinase cleavage site (Ek) downstream from the MBP tag and Xa cleavage site on the original vector. For cloning of a desired peptide DNA, the enterokinase site contains a unique BsaBI restriction site adjacent to the original multi-cloning site. This redesigned pMAL vector was optimized for the purification of cytoplasmic (pMALc5HisEk) and periplasmic (pMALp5HisEk) peptides. The purification of native and active peptide (P) was obtained following two-step affinity chromatography. In the first step, the entire MBP-His(6)-Ek-P fusion protein is purified using the Ni-NTA agarose column. This fusion protein was cleaved with active His(6) tagged enterokinase. In the second step, the further purification was performed by column containing the mixture of amylose and Ni-NTA agarose resins. This removes both the MBP-His(6) and His(6)-enterokinase leaving pure native protein in solution. These new vectors and the two-step purification protocol were successfully applied in purification of active native small antimicrobial peptides (AMPs), lactococcin A and human beta-defensin. Conclusions We constructed the improved pMAL expression vectors and established the pipeline and optimal conditions for their use in efficient purification of large amounts of active native small peptides. Significance and Impact of the Study Choice of expression vector impacts on the efficiency of expression and purification of desired proteins. The idea of redesigning pMAL vector was driven by the need for rapid purification of larger amounts of active native AMPs. This newly improved pMAL vector, the cloning strategy, expression conditions and two-step purification protocol represent a unique simple approach which can be applied in every laboratory.",
publisher = "Wiley, Hoboken",
journal = "Journal of Applied Microbiology",
title = "Redesigned pMAL expression vector for easy and fast purification of active native antimicrobial peptides",
pages = "1013-1001",
number = "2",
volume = "133",
doi = "10.1111/jam.15623"
}

Gardijan, L., Miljković, M., Obradović, M., Borović, B., Vukotić, G., Jovanović, G.,& Kojić, M.. (2022). Redesigned pMAL expression vector for easy and fast purification of active native antimicrobial peptides. in Journal of Applied Microbiology
Wiley, Hoboken., 133(2), 1001-1013.
https://doi.org/10.1111/jam.15623

Gardijan L, Miljković M, Obradović M, Borović B, Vukotić G, Jovanović G, Kojić M. Redesigned pMAL expression vector for easy and fast purification of active native antimicrobial peptides. in Journal of Applied Microbiology. 2022;133(2):1001-1013.
doi:10.1111/jam.15623 .

Gardijan, Lazar, Miljković, Marija, Obradović, Mina, Borović, Branka, Vukotić, Goran, Jovanović, Goran, Kojić, Milan, "Redesigned pMAL expression vector for easy and fast purification of active native antimicrobial peptides" in Journal of Applied Microbiology, 133, no. 2 (2022):1001-1013,
https://doi.org/10.1111/jam.15623 . .

TY  - JOUR
AU  - Mirković, Nemanja
AU  - Obradović, Mina
AU  - O'Connor, Paula M.
AU  - Filipić, Brankica
AU  - Jovčić, Branko
AU  - Cotter, Paul D.
AU  - Kojić, Milan
PY  - 2021
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1429
AB  - Screening for producers of potent antimicrobial peptides, resulted in the isolation of Bacillus cereus BGNM1 with strong antimicrobial activity against Listeria monocytogenes. Genome sequence analysis revealed that BGNM1 contains the gene cluster associated with the production of the lantibiotic, thusin, previously identified in B. thuringiensis. Purification of the antimicrobial activity confirmed that strain BGMN1 produces thusin. Both thusin sensitive and resistant strains were detected among clinical isolates of Streptococcus agalactiae. Random mutagenesis of a thusin sensitive strain, S. agalactiae B782, was performed in an attempt to identify the receptor protein for thusin. Three independent thusin resistant mutants were selected and their complete genomes sequenced. Comparative sequence analysis of these mutants with the WT strain revealed that duplication of a region encoding a 79 amino acids repeat in a C-protein alpha-antigen was a common difference, suggesting it to be responsible for increased resistance to thusin. Since induced thusin resistant mutants showed higher level of resistance than the naturally resistant B761 strain, complete genome sequencing of strain B761 was performed to check the integrity of the C-protein alpha-antigen-encoding gene. This analysis revealed that this gene is deleted in B761, providing further evidence that this protein promotes interaction of the thusin with receptor.
PB  - Springer, Dordrecht
T2  - Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology
T1  - C-protein alpha-antigen modulates the lantibiotic thusin resistance in Streptococcus agalactiae
EP  - 1607
IS  - 10
SP  - 1595
VL  - 114
DO  - 10.1007/s10482-021-01626-3
ER  - 

@article{
author = "Mirković, Nemanja and Obradović, Mina and O'Connor, Paula M. and Filipić, Brankica and Jovčić, Branko and Cotter, Paul D. and Kojić, Milan",
year = "2021",
abstract = "Screening for producers of potent antimicrobial peptides, resulted in the isolation of Bacillus cereus BGNM1 with strong antimicrobial activity against Listeria monocytogenes. Genome sequence analysis revealed that BGNM1 contains the gene cluster associated with the production of the lantibiotic, thusin, previously identified in B. thuringiensis. Purification of the antimicrobial activity confirmed that strain BGMN1 produces thusin. Both thusin sensitive and resistant strains were detected among clinical isolates of Streptococcus agalactiae. Random mutagenesis of a thusin sensitive strain, S. agalactiae B782, was performed in an attempt to identify the receptor protein for thusin. Three independent thusin resistant mutants were selected and their complete genomes sequenced. Comparative sequence analysis of these mutants with the WT strain revealed that duplication of a region encoding a 79 amino acids repeat in a C-protein alpha-antigen was a common difference, suggesting it to be responsible for increased resistance to thusin. Since induced thusin resistant mutants showed higher level of resistance than the naturally resistant B761 strain, complete genome sequencing of strain B761 was performed to check the integrity of the C-protein alpha-antigen-encoding gene. This analysis revealed that this gene is deleted in B761, providing further evidence that this protein promotes interaction of the thusin with receptor.",
publisher = "Springer, Dordrecht",
journal = "Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology",
title = "C-protein alpha-antigen modulates the lantibiotic thusin resistance in Streptococcus agalactiae",
pages = "1607-1595",
number = "10",
volume = "114",
doi = "10.1007/s10482-021-01626-3"
}

Mirković, N., Obradović, M., O'Connor, P. M., Filipić, B., Jovčić, B., Cotter, P. D.,& Kojić, M.. (2021). C-protein alpha-antigen modulates the lantibiotic thusin resistance in Streptococcus agalactiae. in Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology
Springer, Dordrecht., 114(10), 1595-1607.
https://doi.org/10.1007/s10482-021-01626-3

Mirković N, Obradović M, O'Connor PM, Filipić B, Jovčić B, Cotter PD, Kojić M. C-protein alpha-antigen modulates the lantibiotic thusin resistance in Streptococcus agalactiae. in Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology. 2021;114(10):1595-1607.
doi:10.1007/s10482-021-01626-3 .

Mirković, Nemanja, Obradović, Mina, O'Connor, Paula M., Filipić, Brankica, Jovčić, Branko, Cotter, Paul D., Kojić, Milan, "C-protein alpha-antigen modulates the lantibiotic thusin resistance in Streptococcus agalactiae" in Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology, 114, no. 10 (2021):1595-1607,
https://doi.org/10.1007/s10482-021-01626-3 . .

TY  - JOUR
AU  - Vukotić, Goran
AU  - Obradović, Mina
AU  - Novović, Katarina
AU  - Di Luca, Mariagrazia
AU  - Jovčić, Branko
AU  - Fira, Đorđe
AU  - Neve, Horst
AU  - Kojić, Milan
AU  - McAuliffe, Olivia
PY  - 2020
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1355
AB  - Acinetobacter baumanniiis a leading cause of healthcare-associated infections worldwide. Its various intrinsic and acquired mechanisms of antibiotic resistance make the therapeutic challenge even more serious. One of the promising alternative treatments that is increasingly highlighted is phage therapy, the therapeutic use of bacteriophages to treat bacterial infections. Two phages active against nosocomial carbapenem-resistantA. baumanniistrain 6077/12, vB_AbaM_ISTD, and vB_AbaM_NOVI, were isolated from Belgrade wastewaters, purified, and concentrated using CsCl gradient ultracentrifugation. The phages were screened against 103 clinical isolates ofA. baumanniifrom a laboratory collection and characterized based on plaque and virion morphology, host range, adsorption rate, and one-step growth curve. Given that phage ISTD showed a broader host range, better adsorption rate, shorter latent period, and larger burst size, its ability to lyse planktonic and biofilm-embedded cells was tested in detail. Phage ISTD yielded a 3.5- and 2-log reduction in planktonic and biofilm-associated viable bacterial cell count, respectively, but the effect was time-dependent. Both phages produced growing turbid halos around plaques indicating the synthesis of depolymerases, enzymes capable of degrading bacterial exopolysaccharides. Halos tested positive for presence of phages in the proximity of the plaque, but not further from the plaque, which indicates that the observed halo enlargement is a consequence of enzyme diffusion through the agar, independently of the phages. This notion was also supported by the growing halos induced by phage preparations applied on pregrown bacterial lawns, indicating that depolymerizing effect was achieved also on non-dividing sensitive cells. Overall, good rates of growth, fast adsorption rate, broad host range, and high depolymerizing activity, as well as antibacterial effectiveness against planktonic and biofilm-associated bacteria, make these phages good candidates for potential application in combatingA. baumanniiinfections.
PB  - Frontiers Media Sa, Lausanne
T2  - Frontiers in Medicine
T1  - Characterization, Antibiofilm, and Depolymerizing Activity of Two Phages Active on Carbapenem-ResistantAcinetobacter baumannii
VL  - 7
DO  - 10.3389/fmed.2020.00426
ER  - 

@article{
author = "Vukotić, Goran and Obradović, Mina and Novović, Katarina and Di Luca, Mariagrazia and Jovčić, Branko and Fira, Đorđe and Neve, Horst and Kojić, Milan and McAuliffe, Olivia",
year = "2020",
abstract = "Acinetobacter baumanniiis a leading cause of healthcare-associated infections worldwide. Its various intrinsic and acquired mechanisms of antibiotic resistance make the therapeutic challenge even more serious. One of the promising alternative treatments that is increasingly highlighted is phage therapy, the therapeutic use of bacteriophages to treat bacterial infections. Two phages active against nosocomial carbapenem-resistantA. baumanniistrain 6077/12, vB_AbaM_ISTD, and vB_AbaM_NOVI, were isolated from Belgrade wastewaters, purified, and concentrated using CsCl gradient ultracentrifugation. The phages were screened against 103 clinical isolates ofA. baumanniifrom a laboratory collection and characterized based on plaque and virion morphology, host range, adsorption rate, and one-step growth curve. Given that phage ISTD showed a broader host range, better adsorption rate, shorter latent period, and larger burst size, its ability to lyse planktonic and biofilm-embedded cells was tested in detail. Phage ISTD yielded a 3.5- and 2-log reduction in planktonic and biofilm-associated viable bacterial cell count, respectively, but the effect was time-dependent. Both phages produced growing turbid halos around plaques indicating the synthesis of depolymerases, enzymes capable of degrading bacterial exopolysaccharides. Halos tested positive for presence of phages in the proximity of the plaque, but not further from the plaque, which indicates that the observed halo enlargement is a consequence of enzyme diffusion through the agar, independently of the phages. This notion was also supported by the growing halos induced by phage preparations applied on pregrown bacterial lawns, indicating that depolymerizing effect was achieved also on non-dividing sensitive cells. Overall, good rates of growth, fast adsorption rate, broad host range, and high depolymerizing activity, as well as antibacterial effectiveness against planktonic and biofilm-associated bacteria, make these phages good candidates for potential application in combatingA. baumanniiinfections.",
publisher = "Frontiers Media Sa, Lausanne",
journal = "Frontiers in Medicine",
title = "Characterization, Antibiofilm, and Depolymerizing Activity of Two Phages Active on Carbapenem-ResistantAcinetobacter baumannii",
volume = "7",
doi = "10.3389/fmed.2020.00426"
}

Vukotić, G., Obradović, M., Novović, K., Di Luca, M., Jovčić, B., Fira, Đ., Neve, H., Kojić, M.,& McAuliffe, O.. (2020). Characterization, Antibiofilm, and Depolymerizing Activity of Two Phages Active on Carbapenem-ResistantAcinetobacter baumannii. in Frontiers in Medicine
Frontiers Media Sa, Lausanne., 7.
https://doi.org/10.3389/fmed.2020.00426

Vukotić G, Obradović M, Novović K, Di Luca M, Jovčić B, Fira Đ, Neve H, Kojić M, McAuliffe O. Characterization, Antibiofilm, and Depolymerizing Activity of Two Phages Active on Carbapenem-ResistantAcinetobacter baumannii. in Frontiers in Medicine. 2020;7.
doi:10.3389/fmed.2020.00426 .

Vukotić, Goran, Obradović, Mina, Novović, Katarina, Di Luca, Mariagrazia, Jovčić, Branko, Fira, Đorđe, Neve, Horst, Kojić, Milan, McAuliffe, Olivia, "Characterization, Antibiofilm, and Depolymerizing Activity of Two Phages Active on Carbapenem-ResistantAcinetobacter baumannii" in Frontiers in Medicine, 7 (2020),
https://doi.org/10.3389/fmed.2020.00426 . .