TY  - JOUR
AU  - Ćurčić, Jovana
AU  - Dinić, Miroslav
AU  - Novović, Katarina
AU  - Vasiljević, Zorica
AU  - Kojić, Milan
AU  - Jovčić, Branko
AU  - Malešević, Milka
PY  - 2024
UR  - https://www.sciencedirect.com/science/article/pii/S0141813024012248
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2326
AB  - Infections caused by multidrug-resistant pathogens are one of the biggest challenges facing the healthcare system today. Quorum quenching (QQ) enzymes have the potential to be used as innovative enzyme-based antivirulence therapeutics to combat infections caused by multidrug-resistant pathogens. The main objective of this research was to describe the novel YtnP lactonase derived from the clinical isolate Stenotrophomonas maltophilia and to investigate its antivirulence potential against multidrug-resistant Pseudomonas aeruginosa MMA83. YtnP lactonase, the QQ enzyme, belongs to the family of metallo-β-lactamases. The recombinant enzyme has several advantageous biotechnological properties, such as high thermostability, activity in a wide pH range, and no cytotoxic effect. High-performance liquid chromatography analysis revealed the activity of recombinant YtnP lactonase toward a wide range of N-acyl-homoserine lactones (AHLs), quorum sensing signaling molecules, with a higher preference for long-chain AHLs. Recombinant YtnP lactonase was shown to inhibit P. aeruginosa MMA83 biofilm formation, induce biofilm decomposition, and reduce extracellular virulence factors production. Moreover, the lifespan of MMA83-infected Caenorhabditis elegans was prolonged with YtnP lactonase treatment. YtnP lactonase showed synergistic inhibitory activity in combination with gentamicin and acted additively with meropenem against MMA83. The described properties make YtnP lactonase a promising therapeutic candidate for the development of next-generation antivirulence agents.
PB  - Elsevier
T2  - International Journal of Biological Macromolecules
T2  - International Journal of Biological MacromoleculesInternational Journal of Biological Macromolecules
T1  - A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo
SP  - 130421
DO  - 10.1016/j.ijbiomac.2024.130421
ER  - 

@article{
author = "Ćurčić, Jovana and Dinić, Miroslav and Novović, Katarina and Vasiljević, Zorica and Kojić, Milan and Jovčić, Branko and Malešević, Milka",
year = "2024",
abstract = "Infections caused by multidrug-resistant pathogens are one of the biggest challenges facing the healthcare system today. Quorum quenching (QQ) enzymes have the potential to be used as innovative enzyme-based antivirulence therapeutics to combat infections caused by multidrug-resistant pathogens. The main objective of this research was to describe the novel YtnP lactonase derived from the clinical isolate Stenotrophomonas maltophilia and to investigate its antivirulence potential against multidrug-resistant Pseudomonas aeruginosa MMA83. YtnP lactonase, the QQ enzyme, belongs to the family of metallo-β-lactamases. The recombinant enzyme has several advantageous biotechnological properties, such as high thermostability, activity in a wide pH range, and no cytotoxic effect. High-performance liquid chromatography analysis revealed the activity of recombinant YtnP lactonase toward a wide range of N-acyl-homoserine lactones (AHLs), quorum sensing signaling molecules, with a higher preference for long-chain AHLs. Recombinant YtnP lactonase was shown to inhibit P. aeruginosa MMA83 biofilm formation, induce biofilm decomposition, and reduce extracellular virulence factors production. Moreover, the lifespan of MMA83-infected Caenorhabditis elegans was prolonged with YtnP lactonase treatment. YtnP lactonase showed synergistic inhibitory activity in combination with gentamicin and acted additively with meropenem against MMA83. The described properties make YtnP lactonase a promising therapeutic candidate for the development of next-generation antivirulence agents.",
publisher = "Elsevier",
journal = "International Journal of Biological Macromolecules, International Journal of Biological MacromoleculesInternational Journal of Biological Macromolecules",
title = "A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo",
pages = "130421",
doi = "10.1016/j.ijbiomac.2024.130421"
}

Ćurčić, J., Dinić, M., Novović, K., Vasiljević, Z., Kojić, M., Jovčić, B.,& Malešević, M.. (2024). A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo. in International Journal of Biological Macromolecules
Elsevier., 130421.
https://doi.org/10.1016/j.ijbiomac.2024.130421

Ćurčić J, Dinić M, Novović K, Vasiljević Z, Kojić M, Jovčić B, Malešević M. A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo. in International Journal of Biological Macromolecules. 2024;:130421.
doi:10.1016/j.ijbiomac.2024.130421 .

Ćurčić, Jovana, Dinić, Miroslav, Novović, Katarina, Vasiljević, Zorica, Kojić, Milan, Jovčić, Branko, Malešević, Milka, "A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo" in International Journal of Biological Macromolecules (2024):130421,
https://doi.org/10.1016/j.ijbiomac.2024.130421 . .

TY  - JOUR
AU  - Malešević, Milka
AU  - Gardijan, Lazar
AU  - Miljković, Marija
AU  - O'Connor, Paula M
AU  - Mirković, Nemanja
AU  - Jovčić, Branko
AU  - Cotter, Paul D
AU  - Jovanovic, Goran
AU  - Kojić, Milan
PY  - 2023
UR  - https://doi.org/10.1093/lambio/ovad004
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1828
AB  - Lactic acid bacterium Lactococcus lactis BGBU1-4 produces 43 amino acids (aa) long bacteriocin, lactolisterin BU (LBU), a 5.161 kDa peptide with potent antibacterial activity against many Gram-positive pathogens. In addition, BGBU1-4 produces an additional unknown product of 3.642 kDa with antibacterial activity. Here, we determined that the significant amount of naturally produced LBU breaks down to create a 3.642 kDa truncated form of LBU bacteriocin consisting of 31 N-terminal aa (LBU1-31) that exhibits 12.5% the antibacterial activity of the full-length LBU. We showed that chemically synthesized LBU is stable and 50% less active than native LBU, and so we used the synthetic peptides of LBU and its variants to further study their activities and antibacterial potential. Deletion analysis of LBU revealed that the 24 N-terminal aa of LBU (LBU1-24) are responsible for antibacterial activity, while downstream aa (25–43) determine the species-specific effectiveness of LBU. Although LBU1-31 contains aa 1–24, the truncation at position 31 is predicted to change the structure within aa 15–31 and might impact on antibacterial activity. Intriguingly, whole genome sequencing and genome mining established that BGBU1-4 is abundant in genes that encode potential antibacterials, but produces LBU and its breakdown product LBU1-31 exclusively.
T2  - Letters in Applied Microbiology
T2  - Letters in Applied MicrobiologyLetters in Applied Microbiology
T1  - Exploring the antibacterial potential of Lactococcus lactis subsp. lactis bv. diacetylactis BGBU1-4 by genome mining, bacteriocin gene overexpression, and chemical protein synthesis of lactolisterin BU variants
IS  - 2
SP  - ovad004
VL  - 76
DO  - 10.1093/lambio/ovad004
ER  - 

@article{
author = "Malešević, Milka and Gardijan, Lazar and Miljković, Marija and O'Connor, Paula M and Mirković, Nemanja and Jovčić, Branko and Cotter, Paul D and Jovanovic, Goran and Kojić, Milan",
year = "2023",
abstract = "Lactic acid bacterium Lactococcus lactis BGBU1-4 produces 43 amino acids (aa) long bacteriocin, lactolisterin BU (LBU), a 5.161 kDa peptide with potent antibacterial activity against many Gram-positive pathogens. In addition, BGBU1-4 produces an additional unknown product of 3.642 kDa with antibacterial activity. Here, we determined that the significant amount of naturally produced LBU breaks down to create a 3.642 kDa truncated form of LBU bacteriocin consisting of 31 N-terminal aa (LBU1-31) that exhibits 12.5% the antibacterial activity of the full-length LBU. We showed that chemically synthesized LBU is stable and 50% less active than native LBU, and so we used the synthetic peptides of LBU and its variants to further study their activities and antibacterial potential. Deletion analysis of LBU revealed that the 24 N-terminal aa of LBU (LBU1-24) are responsible for antibacterial activity, while downstream aa (25–43) determine the species-specific effectiveness of LBU. Although LBU1-31 contains aa 1–24, the truncation at position 31 is predicted to change the structure within aa 15–31 and might impact on antibacterial activity. Intriguingly, whole genome sequencing and genome mining established that BGBU1-4 is abundant in genes that encode potential antibacterials, but produces LBU and its breakdown product LBU1-31 exclusively.",
journal = "Letters in Applied Microbiology, Letters in Applied MicrobiologyLetters in Applied Microbiology",
title = "Exploring the antibacterial potential of Lactococcus lactis subsp. lactis bv. diacetylactis BGBU1-4 by genome mining, bacteriocin gene overexpression, and chemical protein synthesis of lactolisterin BU variants",
number = "2",
pages = "ovad004",
volume = "76",
doi = "10.1093/lambio/ovad004"
}

Malešević, M., Gardijan, L., Miljković, M., O'Connor, P. M., Mirković, N., Jovčić, B., Cotter, P. D., Jovanovic, G.,& Kojić, M.. (2023). Exploring the antibacterial potential of Lactococcus lactis subsp. lactis bv. diacetylactis BGBU1-4 by genome mining, bacteriocin gene overexpression, and chemical protein synthesis of lactolisterin BU variants. in Letters in Applied Microbiology, 76(2), ovad004.
https://doi.org/10.1093/lambio/ovad004

Malešević M, Gardijan L, Miljković M, O'Connor PM, Mirković N, Jovčić B, Cotter PD, Jovanovic G, Kojić M. Exploring the antibacterial potential of Lactococcus lactis subsp. lactis bv. diacetylactis BGBU1-4 by genome mining, bacteriocin gene overexpression, and chemical protein synthesis of lactolisterin BU variants. in Letters in Applied Microbiology. 2023;76(2):ovad004.
doi:10.1093/lambio/ovad004 .

Malešević, Milka, Gardijan, Lazar, Miljković, Marija, O'Connor, Paula M, Mirković, Nemanja, Jovčić, Branko, Cotter, Paul D, Jovanovic, Goran, Kojić, Milan, "Exploring the antibacterial potential of Lactococcus lactis subsp. lactis bv. diacetylactis BGBU1-4 by genome mining, bacteriocin gene overexpression, and chemical protein synthesis of lactolisterin BU variants" in Letters in Applied Microbiology, 76, no. 2 (2023):ovad004,
https://doi.org/10.1093/lambio/ovad004 . .

TY  - JOUR
AU  - Malešević, Milka
AU  - Stanisavljević, Nemanja
AU  - Matijašević, Danka
AU  - Ćurčić, Jovana
AU  - Tasić, Vukašin
AU  - Tasić, Srđan
AU  - Kojić, Milan
PY  - 2023
UR  - https://doi.org/10.1007/s00248-023-02242-6
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1893
AB  - The hot spring Vranjska Banja is the hottest spring on the Balkan Peninsula with a water temperature of 63–95 °C and a pH value of 7.1, in situ. According to the physicochemical analysis, Vranjska Banja hot spring belongs to the bicarbonated and sulfated hyperthermal waters. The structures of microbial community of this geothermal spring are still largely unexplored. In order to determine and monitor the diversity of microbiota of the Vranjska Banja hot spring, a comprehensive culture-independent metagenomic analysis was conducted in parallel with a culture-dependent approach for the first time. Microbial profiling using amplicon sequencing analysis revealed the presence of phylogenetically novel taxa, ranging from species to phyla. Cultivation-based methods resulted in the isolation of 17 strains belonging to the genera Anoxybacillus, Bacillus, Geobacillus, and Hydrogenophillus. Whole-genome sequencing of five representative strains was then performed. The genomic characterization and OrthoANI analysis revealed that the Vranjska Banja hot spring harbors phylogenetically novel species of the genus Anoxybacillus, proving its uniqueness. Moreover, these isolates contain stress response genes that enable them to survive in the harsh conditions of the hot springs. The results of the in silico analysis show that most of the sequenced strains have the potential to produce thermostable enzymes (proteases, lipases, amylases, phytase, chitinase, and glucanase) and various antimicrobial molecules that can be of great importance for industrial, agricultural, and biotechnological applications. Finally, this study provides a basis for further research and understanding of the metabolic potential of these microorganisms.
T2  - Microbial Ecology
T2  - Microbial EcologyMicrob Ecol
T1  - Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia
DO  - 10.1007/s00248-023-02242-6
ER  - 

@article{
author = "Malešević, Milka and Stanisavljević, Nemanja and Matijašević, Danka and Ćurčić, Jovana and Tasić, Vukašin and Tasić, Srđan and Kojić, Milan",
year = "2023",
abstract = "The hot spring Vranjska Banja is the hottest spring on the Balkan Peninsula with a water temperature of 63–95 °C and a pH value of 7.1, in situ. According to the physicochemical analysis, Vranjska Banja hot spring belongs to the bicarbonated and sulfated hyperthermal waters. The structures of microbial community of this geothermal spring are still largely unexplored. In order to determine and monitor the diversity of microbiota of the Vranjska Banja hot spring, a comprehensive culture-independent metagenomic analysis was conducted in parallel with a culture-dependent approach for the first time. Microbial profiling using amplicon sequencing analysis revealed the presence of phylogenetically novel taxa, ranging from species to phyla. Cultivation-based methods resulted in the isolation of 17 strains belonging to the genera Anoxybacillus, Bacillus, Geobacillus, and Hydrogenophillus. Whole-genome sequencing of five representative strains was then performed. The genomic characterization and OrthoANI analysis revealed that the Vranjska Banja hot spring harbors phylogenetically novel species of the genus Anoxybacillus, proving its uniqueness. Moreover, these isolates contain stress response genes that enable them to survive in the harsh conditions of the hot springs. The results of the in silico analysis show that most of the sequenced strains have the potential to produce thermostable enzymes (proteases, lipases, amylases, phytase, chitinase, and glucanase) and various antimicrobial molecules that can be of great importance for industrial, agricultural, and biotechnological applications. Finally, this study provides a basis for further research and understanding of the metabolic potential of these microorganisms.",
journal = "Microbial Ecology, Microbial EcologyMicrob Ecol",
title = "Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia",
doi = "10.1007/s00248-023-02242-6"
}

Malešević, M., Stanisavljević, N., Matijašević, D., Ćurčić, J., Tasić, V., Tasić, S.,& Kojić, M.. (2023). Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia. in Microbial Ecology.
https://doi.org/10.1007/s00248-023-02242-6

Malešević M, Stanisavljević N, Matijašević D, Ćurčić J, Tasić V, Tasić S, Kojić M. Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia. in Microbial Ecology. 2023;.
doi:10.1007/s00248-023-02242-6 .

Malešević, Milka, Stanisavljević, Nemanja, Matijašević, Danka, Ćurčić, Jovana, Tasić, Vukašin, Tasić, Srđan, Kojić, Milan, "Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia" in Microbial Ecology (2023),
https://doi.org/10.1007/s00248-023-02242-6 . .

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  - Gardijan, Lazar
AU  - Kojić, Milan
AU  - Jovanović, Goran
AU  - Malešević, Milka
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/804
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2125
AB  - Introduction: Lactolisterin BU (LBU) is a potent bacteriocin derived from Lactococcuslactis subsp. lactisbv. diacetylactis BGBU1-4. It exhibits antimicrobial properties against Gram-positive food spoilage andfoodborne pathogens. This research aimed to explore the impact of amino acid substitution in LBU onits antimicrobial activity by utilizing in silico prediction of LBU’ssecondary structure and amino acid substitutions.Methods: The secondary structure of LBU was predicted using Phyre2 software. Five variants of LBUwere selected and chemically synthesized, along with unaltered LBU and BHT-B,serving as controls. Peptides were twofold diluted in distilled water, resulting in final concentrations ranging from 1000 µg/mlto 0.5 µg/ml. An agarspot test, employing 5 µl of the dilution, was conducted on three indicatorstrains:Lactococcus lactis BGMN1-596, Listeria monocytogenes ATCC19111, and Staphylococcus aureusATCC25923. The presence of inhibition zones was analyzed after overnight incubation at 37°C (S. aureus)and 30°C (L. lactis and L. monocytogenes).Results: Phyre2 analysis unveiled the presence of two α-helices in LBU’s structure. The majority of LBUvariants displayed altered antimicrobial activity, with some changes being genusspecific, potentially attributable to variances in cell wall composition. Some variants completely lost their activity, underscoring the significance of native amino acids or their physicochemical properties in the correspondingpositions within LBU’s structure. Furthermore, it was confirmed that chemically synthesized LBU effectively retains its antimicrobial activity.Conclusion: Changesin amino acid composition give insight on structure-function relationship of LBU.
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  - Influence of amino acid substitution on the antimicrobial activity of bacteriocin lactolisterin BU
EP  - 123
SP  - 123
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2125
ER  - 

@conference{
author = "Gardijan, Lazar and Kojić, Milan and Jovanović, Goran and Malešević, Milka",
year = "2023",
abstract = "Introduction: Lactolisterin BU (LBU) is a potent bacteriocin derived from Lactococcuslactis subsp. lactisbv. diacetylactis BGBU1-4. It exhibits antimicrobial properties against Gram-positive food spoilage andfoodborne pathogens. This research aimed to explore the impact of amino acid substitution in LBU onits antimicrobial activity by utilizing in silico prediction of LBU’ssecondary structure and amino acid substitutions.Methods: The secondary structure of LBU was predicted using Phyre2 software. Five variants of LBUwere selected and chemically synthesized, along with unaltered LBU and BHT-B,serving as controls. Peptides were twofold diluted in distilled water, resulting in final concentrations ranging from 1000 µg/mlto 0.5 µg/ml. An agarspot test, employing 5 µl of the dilution, was conducted on three indicatorstrains:Lactococcus lactis BGMN1-596, Listeria monocytogenes ATCC19111, and Staphylococcus aureusATCC25923. The presence of inhibition zones was analyzed after overnight incubation at 37°C (S. aureus)and 30°C (L. lactis and L. monocytogenes).Results: Phyre2 analysis unveiled the presence of two α-helices in LBU’s structure. The majority of LBUvariants displayed altered antimicrobial activity, with some changes being genusspecific, potentially attributable to variances in cell wall composition. Some variants completely lost their activity, underscoring the significance of native amino acids or their physicochemical properties in the correspondingpositions within LBU’s structure. Furthermore, it was confirmed that chemically synthesized LBU effectively retains its antimicrobial activity.Conclusion: Changesin amino acid composition give insight on structure-function relationship of LBU.",
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 = "Influence of amino acid substitution on the antimicrobial activity of bacteriocin lactolisterin BU",
pages = "123-123",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2125"
}

Gardijan, L., Kojić, M., Jovanović, G.,& Malešević, M.. (2023). Influence of amino acid substitution on the antimicrobial activity of bacteriocin lactolisterin BU. 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., 123-123.
https://hdl.handle.net/21.15107/rcub_imagine_2125

Gardijan L, Kojić M, Jovanović G, Malešević M. Influence of amino acid substitution on the antimicrobial activity of bacteriocin lactolisterin BU. 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;:123-123.
https://hdl.handle.net/21.15107/rcub_imagine_2125 .

Gardijan, Lazar, Kojić, Milan, Jovanović, Goran, Malešević, Milka, "Influence of amino acid substitution on the antimicrobial activity of bacteriocin lactolisterin BU" 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):123-123,
https://hdl.handle.net/21.15107/rcub_imagine_2125 .

TY  - JOUR
AU  - Malešević, Milka
AU  - Stanisavljević, Nemanja
AU  - Matijašević, Danka
AU  - Ćurčić, Jovana
AU  - Tasić, Vukašin
AU  - Tasić, Srđan
AU  - Kojić, Milan
PY  - 2023
UR  - https://doi.org/10.1007/s00248-023-02242-6
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1931
AB  - The hot spring Vranjska Banja is the hottest spring on the Balkan Peninsula with a water temperature of 63–95 °C and a pH value of 7.1, in situ. According to the physicochemical analysis, Vranjska Banja hot spring belongs to the bicarbonated and sulfated hyperthermal waters. The structures of microbial community of this geothermal spring are still largely unexplored. In order to determine and monitor the diversity of microbiota of the Vranjska Banja hot spring, a comprehensive culture-independent metagenomic analysis was conducted in parallel with a culture-dependent approach for the first time. Microbial profiling using amplicon sequencing analysis revealed the presence of phylogenetically novel taxa, ranging from species to phyla. Cultivation-based methods resulted in the isolation of 17 strains belonging to the genera Anoxybacillus, Bacillus, Geobacillus, and Hydrogenophillus. Whole-genome sequencing of five representative strains was then performed. The genomic characterization and OrthoANI analysis revealed that the Vranjska Banja hot spring harbors phylogenetically novel species of the genus Anoxybacillus, proving its uniqueness. Moreover, these isolates contain stress response genes that enable them to survive in the harsh conditions of the hot springs. The results of the in silico analysis show that most of the sequenced strains have the potential to produce thermostable enzymes (proteases, lipases, amylases, phytase, chitinase, and glucanase) and various antimicrobial molecules that can be of great importance for industrial, agricultural, and biotechnological applications. Finally, this study provides a basis for further research and understanding of the metabolic potential of these microorganisms.
T2  - Microbial Ecology
T2  - Microbial EcologyMicrob Ecol
T1  - Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia
DO  - 10.1007/s00248-023-02242-6
ER  - 

@article{
author = "Malešević, Milka and Stanisavljević, Nemanja and Matijašević, Danka and Ćurčić, Jovana and Tasić, Vukašin and Tasić, Srđan and Kojić, Milan",
year = "2023",
abstract = "The hot spring Vranjska Banja is the hottest spring on the Balkan Peninsula with a water temperature of 63–95 °C and a pH value of 7.1, in situ. According to the physicochemical analysis, Vranjska Banja hot spring belongs to the bicarbonated and sulfated hyperthermal waters. The structures of microbial community of this geothermal spring are still largely unexplored. In order to determine and monitor the diversity of microbiota of the Vranjska Banja hot spring, a comprehensive culture-independent metagenomic analysis was conducted in parallel with a culture-dependent approach for the first time. Microbial profiling using amplicon sequencing analysis revealed the presence of phylogenetically novel taxa, ranging from species to phyla. Cultivation-based methods resulted in the isolation of 17 strains belonging to the genera Anoxybacillus, Bacillus, Geobacillus, and Hydrogenophillus. Whole-genome sequencing of five representative strains was then performed. The genomic characterization and OrthoANI analysis revealed that the Vranjska Banja hot spring harbors phylogenetically novel species of the genus Anoxybacillus, proving its uniqueness. Moreover, these isolates contain stress response genes that enable them to survive in the harsh conditions of the hot springs. The results of the in silico analysis show that most of the sequenced strains have the potential to produce thermostable enzymes (proteases, lipases, amylases, phytase, chitinase, and glucanase) and various antimicrobial molecules that can be of great importance for industrial, agricultural, and biotechnological applications. Finally, this study provides a basis for further research and understanding of the metabolic potential of these microorganisms.",
journal = "Microbial Ecology, Microbial EcologyMicrob Ecol",
title = "Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia",
doi = "10.1007/s00248-023-02242-6"
}

Malešević, M., Stanisavljević, N., Matijašević, D., Ćurčić, J., Tasić, V., Tasić, S.,& Kojić, M.. (2023). Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia. in Microbial Ecology.
https://doi.org/10.1007/s00248-023-02242-6

Malešević M, Stanisavljević N, Matijašević D, Ćurčić J, Tasić V, Tasić S, Kojić M. Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia. in Microbial Ecology. 2023;.
doi:10.1007/s00248-023-02242-6 .

Malešević, Milka, Stanisavljević, Nemanja, Matijašević, Danka, Ćurčić, Jovana, Tasić, Vukašin, Tasić, Srđan, Kojić, Milan, "Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia" in Microbial Ecology (2023),
https://doi.org/10.1007/s00248-023-02242-6 . .

TY  - DATA
AU  - Novović, Katarina
AU  - Kuzmanović Nedeljković, Snežana
AU  - Poledica, Mirjana
AU  - Nikolić, Gordana
AU  - Grujić, Bojana
AU  - Jovčić, Branko
AU  - Kojić, Milan
AU  - Filipić, Brankica
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1783
T2  - Frontiers in Microbiology
T2  - Frontiers in Microbiology
T1  - Supplementary information for the article: Novović, K., Kuzmanović Nedeljković, S., Poledica, M., Nikolić, G., Grujić, B., Jovčić, B., Kojić, M., & Filipić, B. (2023). Virulence potential of multidrug-resistant Acinetobacter baumannii isolates from COVID-19 patients on mechanical ventilation: The first report from Serbia. Frontiers in Microbiology, 14. https://www.frontiersin.org/articles/10.3389/fmicb.2023.1094184
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1783
ER  - 

@misc{
author = "Novović, Katarina and Kuzmanović Nedeljković, Snežana and Poledica, Mirjana and Nikolić, Gordana and Grujić, Bojana and Jovčić, Branko and Kojić, Milan and Filipić, Brankica",
year = "2023",
journal = "Frontiers in Microbiology, Frontiers in Microbiology",
title = "Supplementary information for the article: Novović, K., Kuzmanović Nedeljković, S., Poledica, M., Nikolić, G., Grujić, B., Jovčić, B., Kojić, M., & Filipić, B. (2023). Virulence potential of multidrug-resistant Acinetobacter baumannii isolates from COVID-19 patients on mechanical ventilation: The first report from Serbia. Frontiers in Microbiology, 14. https://www.frontiersin.org/articles/10.3389/fmicb.2023.1094184",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1783"
}

Novović, K., Kuzmanović Nedeljković, S., Poledica, M., Nikolić, G., Grujić, B., Jovčić, B., Kojić, M.,& Filipić, B.. (2023). Supplementary information for the article: Novović, K., Kuzmanović Nedeljković, S., Poledica, M., Nikolić, G., Grujić, B., Jovčić, B., Kojić, M., & Filipić, B. (2023). Virulence potential of multidrug-resistant Acinetobacter baumannii isolates from COVID-19 patients on mechanical ventilation: The first report from Serbia. Frontiers in Microbiology, 14. https://www.frontiersin.org/articles/10.3389/fmicb.2023.1094184. in Frontiers in Microbiology.
https://hdl.handle.net/21.15107/rcub_imagine_1783

Novović K, Kuzmanović Nedeljković S, Poledica M, Nikolić G, Grujić B, Jovčić B, Kojić M, Filipić B. Supplementary information for the article: Novović, K., Kuzmanović Nedeljković, S., Poledica, M., Nikolić, G., Grujić, B., Jovčić, B., Kojić, M., & Filipić, B. (2023). Virulence potential of multidrug-resistant Acinetobacter baumannii isolates from COVID-19 patients on mechanical ventilation: The first report from Serbia. Frontiers in Microbiology, 14. https://www.frontiersin.org/articles/10.3389/fmicb.2023.1094184. in Frontiers in Microbiology. 2023;.
https://hdl.handle.net/21.15107/rcub_imagine_1783 .

Novović, Katarina, Kuzmanović Nedeljković, Snežana, Poledica, Mirjana, Nikolić, Gordana, Grujić, Bojana, Jovčić, Branko, Kojić, Milan, Filipić, Brankica, "Supplementary information for the article: Novović, K., Kuzmanović Nedeljković, S., Poledica, M., Nikolić, G., Grujić, B., Jovčić, B., Kojić, M., & Filipić, B. (2023). Virulence potential of multidrug-resistant Acinetobacter baumannii isolates from COVID-19 patients on mechanical ventilation: The first report from Serbia. Frontiers in Microbiology, 14. https://www.frontiersin.org/articles/10.3389/fmicb.2023.1094184" in Frontiers in Microbiology (2023),
https://hdl.handle.net/21.15107/rcub_imagine_1783 .

TY  - CONF
AU  - Ćurčić, Jovana
AU  - Jakovljević, Stefan
AU  - Novović, Katarina
AU  - Vasiljević, Zorica
AU  - Kojić, Milan
AU  - Jovčić, Branko
AU  - Malešević, Milka
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/803
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2124
AB  - Introduction: Quorum quenching (QQ) isthe enzymatic degradation of cell-to-cellsignaling molecules.In this study, the potential of the novel YtnP lactonase, the quorum quenching enzyme derived from S.maltophilia, to reduce P. aeruginosa quorum sensing and virulence factor gene expression was investigated.Methods: MMA83 culture (adjusted to 1.5x105 CFU/ml) was treated with recombinant YtnP lactonase(final concentration 50 μg/ml) at 37°C for 12 hours under aeration. RNA isolation of the treated and untreated MMA83 culture was performed using the RNeasy Mini Kit (Qiagen, Germany) according to theprotocol. Quantitative reverse transcription-polymerase chain reaction (RT-qPCR), was used to analyzethe effect ofYtnP lactonase on the relative mRNA levels of the LasI/LasR, RhiI/RhiR, and PQS signaling network genes of P. aeruginosa MMA83 and virulence factor genes. The rpsL was used as an endogenouscontrol to normalize obtained data following the 2-ΔΔCt method.Results: The QS genes belonging to three QS networks – LasI/LasR, RhiI/RhiR, and PQS of P. aeruginosaMMA83 treated with YtnP lactonase were significantly downregulated. The RT -qPCR results show thattreatment with YtnP-lactonase decreased the relative mRNA levels of genes involved in the productionof elastase (lasB approximately 2-fold), alginate (algK approximately 2.2-fold), pyocyanin (phzM approximately 3.5-fold), pyoverdin (pvdS approximately 2-fold), and rhamnolipid (rhlC approximately 4-fold).These results suggest that YtnP lactonase exerts an antivirulence effect at the transcription level.Conclusion: YtnP lactonase, a quorum quenching (QQ) enzyme, has the potential to be used as an innovative enzyme-based antivirulence therapeutic to combat infections caused by P. aeruginosa.
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  - A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression
EP  - 121
SP  - 121
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2124
ER  - 

@conference{
author = "Ćurčić, Jovana and Jakovljević, Stefan and Novović, Katarina and Vasiljević, Zorica and Kojić, Milan and Jovčić, Branko and Malešević, Milka",
year = "2023",
abstract = "Introduction: Quorum quenching (QQ) isthe enzymatic degradation of cell-to-cellsignaling molecules.In this study, the potential of the novel YtnP lactonase, the quorum quenching enzyme derived from S.maltophilia, to reduce P. aeruginosa quorum sensing and virulence factor gene expression was investigated.Methods: MMA83 culture (adjusted to 1.5x105 CFU/ml) was treated with recombinant YtnP lactonase(final concentration 50 μg/ml) at 37°C for 12 hours under aeration. RNA isolation of the treated and untreated MMA83 culture was performed using the RNeasy Mini Kit (Qiagen, Germany) according to theprotocol. Quantitative reverse transcription-polymerase chain reaction (RT-qPCR), was used to analyzethe effect ofYtnP lactonase on the relative mRNA levels of the LasI/LasR, RhiI/RhiR, and PQS signaling network genes of P. aeruginosa MMA83 and virulence factor genes. The rpsL was used as an endogenouscontrol to normalize obtained data following the 2-ΔΔCt method.Results: The QS genes belonging to three QS networks – LasI/LasR, RhiI/RhiR, and PQS of P. aeruginosaMMA83 treated with YtnP lactonase were significantly downregulated. The RT -qPCR results show thattreatment with YtnP-lactonase decreased the relative mRNA levels of genes involved in the productionof elastase (lasB approximately 2-fold), alginate (algK approximately 2.2-fold), pyocyanin (phzM approximately 3.5-fold), pyoverdin (pvdS approximately 2-fold), and rhamnolipid (rhlC approximately 4-fold).These results suggest that YtnP lactonase exerts an antivirulence effect at the transcription level.Conclusion: YtnP lactonase, a quorum quenching (QQ) enzyme, has the potential to be used as an innovative enzyme-based antivirulence therapeutic to combat infections caused by P. aeruginosa.",
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 = "A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression",
pages = "121-121",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2124"
}

Ćurčić, J., Jakovljević, S., Novović, K., Vasiljević, Z., Kojić, M., Jovčić, B.,& Malešević, M.. (2023). A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression. 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., 121-121.
https://hdl.handle.net/21.15107/rcub_imagine_2124

Ćurčić J, Jakovljević S, Novović K, Vasiljević Z, Kojić M, Jovčić B, Malešević M. A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression. 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;:121-121.
https://hdl.handle.net/21.15107/rcub_imagine_2124 .

Ćurčić, Jovana, Jakovljević, Stefan, Novović, Katarina, Vasiljević, Zorica, Kojić, Milan, Jovčić, Branko, Malešević, Milka, "A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression" 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):121-121,
https://hdl.handle.net/21.15107/rcub_imagine_2124 .

TY  - JOUR
AU  - Novović, Katarina
AU  - Kuzmanović Nedeljković, Snežana
AU  - Poledica, Mirjana
AU  - Nikolić, Gordana
AU  - Grujić, Bojana
AU  - Jovčić, Branko
AU  - Kojić, Milan
AU  - Filipić, Brankica
PY  - 2023
UR  - https://www.frontiersin.org/articles/10.3389/fmicb.2023.1094184
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1782
AB  - Since the WHO declared the COVID-19 pandemic in March 2020, the disease has spread rapidly leading to overload of the health system and many of the patients infected with SARS-CoV-2 needed to be admitted to the intensive care unit (ICU). Around 10% of patients with the severe manifestation of COVID-19 need noninvasive or invasive mechanical ventilation, which represent a risk factor for Acinetobacter baumannii superinfection. The 64 A. baumannii isolates were recovered from COVID-19 patients admitted to ICU at General Hospital “Dr Laza K. Lazarević” Šabac, Serbia, during the period from December 2020 to February 2021. All patients required mechanical ventilation and mortality rate was 100%. The goal of this study was to evaluate antibiotic resistance profiles and virulence potential of A. baumannii isolates recovered from patients with severe form of COVID-19 who had a need for mechanical ventilation. All tested A. baumannii isolates (n = 64) were sensitive to colistin, while resistant to meropenem, imipenem, gentamicin, tobramycin, and levofloxacin according to the broth microdilution method and MDR phenotype was confirmed. In all tested isolates, representatives of international clone 2 (IC2) classified by multiplex PCR for clonal lineage identification, blaAmpC, blaOXA-51, and blaOXA-23 genes were present, as well as ISAba1 insertion sequence upstream of blaOXA-23. Clonal distribution of one dominant strain was found, but individual strains showed phenotypic differences in the level of antibiotic resistance, biofilm formation, and binding to mucin and motility. According to PFGE, four isolates were sequenced and antibiotic resistance genes as well as virulence factors genes were analyzed in these genomes. The results of this study represent the first report on virulence potential of MDR A. baumannii from hospital in Serbia.
T2  - Frontiers in Microbiology
T2  - Frontiers in Microbiology
T1  - Virulence potential of multidrug-resistant Acinetobacter baumannii isolates from COVID-19 patients on mechanical ventilation: The first report from Serbia
VL  - 14
DO  - 10.3389/fmicb.2023.1094184
ER  - 

@article{
author = "Novović, Katarina and Kuzmanović Nedeljković, Snežana and Poledica, Mirjana and Nikolić, Gordana and Grujić, Bojana and Jovčić, Branko and Kojić, Milan and Filipić, Brankica",
year = "2023",
abstract = "Since the WHO declared the COVID-19 pandemic in March 2020, the disease has spread rapidly leading to overload of the health system and many of the patients infected with SARS-CoV-2 needed to be admitted to the intensive care unit (ICU). Around 10% of patients with the severe manifestation of COVID-19 need noninvasive or invasive mechanical ventilation, which represent a risk factor for Acinetobacter baumannii superinfection. The 64 A. baumannii isolates were recovered from COVID-19 patients admitted to ICU at General Hospital “Dr Laza K. Lazarević” Šabac, Serbia, during the period from December 2020 to February 2021. All patients required mechanical ventilation and mortality rate was 100%. The goal of this study was to evaluate antibiotic resistance profiles and virulence potential of A. baumannii isolates recovered from patients with severe form of COVID-19 who had a need for mechanical ventilation. All tested A. baumannii isolates (n = 64) were sensitive to colistin, while resistant to meropenem, imipenem, gentamicin, tobramycin, and levofloxacin according to the broth microdilution method and MDR phenotype was confirmed. In all tested isolates, representatives of international clone 2 (IC2) classified by multiplex PCR for clonal lineage identification, blaAmpC, blaOXA-51, and blaOXA-23 genes were present, as well as ISAba1 insertion sequence upstream of blaOXA-23. Clonal distribution of one dominant strain was found, but individual strains showed phenotypic differences in the level of antibiotic resistance, biofilm formation, and binding to mucin and motility. According to PFGE, four isolates were sequenced and antibiotic resistance genes as well as virulence factors genes were analyzed in these genomes. The results of this study represent the first report on virulence potential of MDR A. baumannii from hospital in Serbia.",
journal = "Frontiers in Microbiology, Frontiers in Microbiology",
title = "Virulence potential of multidrug-resistant Acinetobacter baumannii isolates from COVID-19 patients on mechanical ventilation: The first report from Serbia",
volume = "14",
doi = "10.3389/fmicb.2023.1094184"
}

Novović, K., Kuzmanović Nedeljković, S., Poledica, M., Nikolić, G., Grujić, B., Jovčić, B., Kojić, M.,& Filipić, B.. (2023). Virulence potential of multidrug-resistant Acinetobacter baumannii isolates from COVID-19 patients on mechanical ventilation: The first report from Serbia. in Frontiers in Microbiology, 14.
https://doi.org/10.3389/fmicb.2023.1094184

Novović K, Kuzmanović Nedeljković S, Poledica M, Nikolić G, Grujić B, Jovčić B, Kojić M, Filipić B. Virulence potential of multidrug-resistant Acinetobacter baumannii isolates from COVID-19 patients on mechanical ventilation: The first report from Serbia. in Frontiers in Microbiology. 2023;14.
doi:10.3389/fmicb.2023.1094184 .

Novović, Katarina, Kuzmanović Nedeljković, Snežana, Poledica, Mirjana, Nikolić, Gordana, Grujić, Bojana, Jovčić, Branko, Kojić, Milan, Filipić, Brankica, "Virulence potential of multidrug-resistant Acinetobacter baumannii isolates from COVID-19 patients on mechanical ventilation: The first report from Serbia" in Frontiers in Microbiology, 14 (2023),
https://doi.org/10.3389/fmicb.2023.1094184 . .

TY  - CONF
AU  - Curčić, Jovana
AU  - Matijasević, Danka
AU  - Stanisavljević, Nemanja
AU  - Tasić, Srđan
AU  - Kojić, Milan
AU  - Malešević, Milka
PY  - 2023
UR  - https://belbi.bg.ac.rs/
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2057
AB  - Thermal springs represent a habitat with extreme conditions that harbor a unique
microbial community adapted to thrive in this environment. In addition to the geothermal
springs in Iceland, the thermal springs of Vranjska Banja are considered the hottest in
Europe with a water temperature of 63-95°C. Due to global warming and climate change,
there is a growing need for knowledge about the biodiversity of extreme natural habitats.
Besides the exceptional importance of studying extremophilic microorganisms, the
difficulty in their cultivation limits the expanding necessity of research in this field. This
study provides information about the microbial community structure and physicochemical
characteristics of the thermal spring of Vranjska Banja. To determine and monitor the
microbiota diversity of the Vranjska Banja hot spring, for the first time, comprehensive
culture-independent metagenomic analysis in parallel with a culture-dependent
approach was applied. The culture-independent composition of bacterial communities of
the thermal water was investigated using MiSeq-Illumina technology and analyzed by
the computing environment QIIME2 v2021. The applied cultivation approach resulted
in the isolation of 17 strains belonging to genera Bacillus, Anoxybacillus, Hydrogenophilus,
and Geobacillus, based on 16S rRNA sequencing and whole genome sequencing of five
representative strains has been performed. The complete DNA was sequenced using
Illumina HiSeq from the MicrobesNG service. Genomic characterization and OrthoANI
analysis have shown that two of them are candidates for novel species. Products of
extremophilic microorganisms adapted to harsh conditions have great potential to be
used for biotechnological research and industrial application. Results of BAGEL4 and
AntiSMASH showed that the sequenced strains from Vranjska Banja hot spring have
the potential to produce thermostable enzymes (proteases, lipases, amylases, phytase,
chitinase, and glucanase) and various antimicrobial molecules.
PB  - Belgrade : Institute of molecular genetics and genetic engineering
C3  - 4th Belgrade Bioinformatics Conference
T1  - Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia
EP  - 112
SP  - 112
VL  - 4
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2057
ER  - 

@conference{
author = "Curčić, Jovana and Matijasević, Danka and Stanisavljević, Nemanja and Tasić, Srđan and Kojić, Milan and Malešević, Milka",
year = "2023",
abstract = "Thermal springs represent a habitat with extreme conditions that harbor a unique
microbial community adapted to thrive in this environment. In addition to the geothermal
springs in Iceland, the thermal springs of Vranjska Banja are considered the hottest in
Europe with a water temperature of 63-95°C. Due to global warming and climate change,
there is a growing need for knowledge about the biodiversity of extreme natural habitats.
Besides the exceptional importance of studying extremophilic microorganisms, the
difficulty in their cultivation limits the expanding necessity of research in this field. This
study provides information about the microbial community structure and physicochemical
characteristics of the thermal spring of Vranjska Banja. To determine and monitor the
microbiota diversity of the Vranjska Banja hot spring, for the first time, comprehensive
culture-independent metagenomic analysis in parallel with a culture-dependent
approach was applied. The culture-independent composition of bacterial communities of
the thermal water was investigated using MiSeq-Illumina technology and analyzed by
the computing environment QIIME2 v2021. The applied cultivation approach resulted
in the isolation of 17 strains belonging to genera Bacillus, Anoxybacillus, Hydrogenophilus,
and Geobacillus, based on 16S rRNA sequencing and whole genome sequencing of five
representative strains has been performed. The complete DNA was sequenced using
Illumina HiSeq from the MicrobesNG service. Genomic characterization and OrthoANI
analysis have shown that two of them are candidates for novel species. Products of
extremophilic microorganisms adapted to harsh conditions have great potential to be
used for biotechnological research and industrial application. Results of BAGEL4 and
AntiSMASH showed that the sequenced strains from Vranjska Banja hot spring have
the potential to produce thermostable enzymes (proteases, lipases, amylases, phytase,
chitinase, and glucanase) and various antimicrobial molecules.",
publisher = "Belgrade : Institute of molecular genetics and genetic engineering",
journal = "4th Belgrade Bioinformatics Conference",
title = "Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia",
pages = "112-112",
volume = "4",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2057"
}

Curčić, J., Matijasević, D., Stanisavljević, N., Tasić, S., Kojić, M.,& Malešević, M.. (2023). Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia. in 4th Belgrade Bioinformatics Conference
Belgrade : Institute of molecular genetics and genetic engineering., 4, 112-112.
https://hdl.handle.net/21.15107/rcub_imagine_2057

Curčić J, Matijasević D, Stanisavljević N, Tasić S, Kojić M, Malešević M. Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia. in 4th Belgrade Bioinformatics Conference. 2023;4:112-112.
https://hdl.handle.net/21.15107/rcub_imagine_2057 .

Curčić, Jovana, Matijasević, Danka, Stanisavljević, Nemanja, Tasić, Srđan, Kojić, Milan, Malešević, Milka, "Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia" in 4th Belgrade Bioinformatics Conference, 4 (2023):112-112,
https://hdl.handle.net/21.15107/rcub_imagine_2057 .

TY  - JOUR
AU  - Slavić, Marinela Šokarda
AU  - Kojić, Milan
AU  - Margetić, Aleksandra
AU  - Stanisavljević, Nemanja
AU  - Gardijan, Lazar
AU  - Božić, Nataša
AU  - Vujčić, Zoran
PY  - 2023
UR  - https://www.sciencedirect.com/science/article/pii/S0141813023029501
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2066
AB  - α-Amylase from the thermophilic bacterial strain Anoxybacillus vranjensis ST4 (AVA) was cloned into the pMALc5HisEk expression vector and successfully expressed and purified from the Escherichia coli ER2523 host strain. AVA belongs to the GH13_5 subfamily of glycoside hydrolases and has 7 conserved sequence regions (CSRs) distributed in three distinct domains (A, B, C). In addition, there is a starch binding domain (SBD) from the CBM20 family of carbohydrate binding modules (CBMs). AVA is a monomer of 66 kDa that achieves maximum activity at 60–80 °C and is active and stable over a wide pH range (4.0–9.0). AVA retained 50 % of its activity after 31 h of incubation at 60 °C and was resistant to a large number of denaturing agents. It hydrolyzed starch granules very efficiently, releasing maltose, maltotriose and maltopentaose as the main products. The hydrolysis rates of raw corn, wheat, horseradish, and potato starch, at a concentration of 10 %, were 87.8, 85.9, 93.0, and 58 %, respectively, at pH 8.5 over a 3 h period. This study showed that the high level of expression as well as the properties of this highly stable and versatile enzyme show all the prerequisites for successful application in industry.
T2  - International Journal of Biological Macromolecules
T1  - Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications
SP  - 126055
VL  - 249
DO  - 10.1016/j.ijbiomac.2023.126055
ER  - 

@article{
author = "Slavić, Marinela Šokarda and Kojić, Milan and Margetić, Aleksandra and Stanisavljević, Nemanja and Gardijan, Lazar and Božić, Nataša and Vujčić, Zoran",
year = "2023",
abstract = "α-Amylase from the thermophilic bacterial strain Anoxybacillus vranjensis ST4 (AVA) was cloned into the pMALc5HisEk expression vector and successfully expressed and purified from the Escherichia coli ER2523 host strain. AVA belongs to the GH13_5 subfamily of glycoside hydrolases and has 7 conserved sequence regions (CSRs) distributed in three distinct domains (A, B, C). In addition, there is a starch binding domain (SBD) from the CBM20 family of carbohydrate binding modules (CBMs). AVA is a monomer of 66 kDa that achieves maximum activity at 60–80 °C and is active and stable over a wide pH range (4.0–9.0). AVA retained 50 % of its activity after 31 h of incubation at 60 °C and was resistant to a large number of denaturing agents. It hydrolyzed starch granules very efficiently, releasing maltose, maltotriose and maltopentaose as the main products. The hydrolysis rates of raw corn, wheat, horseradish, and potato starch, at a concentration of 10 %, were 87.8, 85.9, 93.0, and 58 %, respectively, at pH 8.5 over a 3 h period. This study showed that the high level of expression as well as the properties of this highly stable and versatile enzyme show all the prerequisites for successful application in industry.",
journal = "International Journal of Biological Macromolecules",
title = "Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications",
pages = "126055",
volume = "249",
doi = "10.1016/j.ijbiomac.2023.126055"
}

Slavić, M. Š., Kojić, M., Margetić, A., Stanisavljević, N., Gardijan, L., Božić, N.,& Vujčić, Z.. (2023). Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications. in International Journal of Biological Macromolecules, 249, 126055.
https://doi.org/10.1016/j.ijbiomac.2023.126055

Slavić MŠ, Kojić M, Margetić A, Stanisavljević N, Gardijan L, Božić N, Vujčić Z. Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications. in International Journal of Biological Macromolecules. 2023;249:126055.
doi:10.1016/j.ijbiomac.2023.126055 .

Slavić, Marinela Šokarda, Kojić, Milan, Margetić, Aleksandra, Stanisavljević, Nemanja, Gardijan, Lazar, Božić, Nataša, Vujčić, Zoran, "Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications" in International Journal of Biological Macromolecules, 249 (2023):126055,
https://doi.org/10.1016/j.ijbiomac.2023.126055 . .

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  - CONF
AU  - Simonović, Ana
AU  - Filipović, Biljana
AU  - Banjanac, Tijana
AU  - Kojić, Milan
AU  - Ćuković, Katarina
AU  - Subotić, Angelina
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1743
AB  - Biljka kičica (Centaurium erythraea) je interesantna ne samo zbog svojih
lekovitih svojstava, već i zbog izuzetne razvojne plastičnosti i regenerative
sposobnosti u in vitro kulturi. Kako bi identifikovali gene uključene u
morfogenetske procese, sekundarni metabolizam i odgovore na stres, pored
sekvenciranih transkriptoma1 je potreban i sekvenciran genom. Za sekvenciranje
genoma najmodernijom PacBio tehnologijom, neophodna je veoma čista
visokomolekularna DNK, čijih 90% čine lanci > 20 kb. DNK je izolovana iz
mladih listova diploidnog varijeteta kičice gajene in vitro, korišćenjem četiri
različita protokola: (1) Quick-DNA HMW MagBead Kit (Zymo Research, #D6060);
(2) modifikovani CTAB protokol;2 (3) protokol za izolaciju bakterijske DNK
prilagođen biljnim tkivima i (4) izmenjen protokol za izolaciju RNK iz bora.3
Kvalitet DNK izolata je proveravan spektrofotometrijski Nano Drop aparatom,
običnom elektroforezom, pulsnom elektroforezom i Femto Pulsnom analizom.
Protokol (1) daje veoma niske prinose, bilo kad se koristi samostalno ili kao
vid prečišćavanja DNK izolovane na drugi način. Protokolom (3) se dobija visok
prinos DNK nezadovoljavajućeg kvaliteta. Najkvalitetniju DNK u količinama
dovoljnim za sekvenciranje su dali protokoli (2) i (4).
AB  - Биљка кичица (Centaurium erythraea) је интересантна не само због својих
лековитих својстава, већ и због изузетне развојне пластичности и регенеративе
способности у in vitro култури. Како би идентификовали гене укључене у
морфогенетске процесе, секундарни метаболизам и одговоре на стрес, поред
секвенцираних транскриптома1 је потребан и секвенциран геном. За секвенцирање
генома најмодернијом PacBio технологијом, неопходна је веома чиста
високомолекуларна ДНК, чијих 90% чине ланци > 20 kb. ДНК је изолована из
младих листова диплоидног варијетета кичице гајене in vitro, коришћењем четири
различита протокола: (1) Quick-DNA HMW MagBead Kit (Zymo Research, #D6060);
(2) модификовани ЦТАБ протокол;2 (3) протокол за изолацију бактеријске ДНК
прилагођен биљним ткивима и (4) измењен протокол за изолацију РНК из бора.3
Квалитет ДНК изолата је провераван спектрофотометријски Nano Drop апаратом,
обичном електрофорезом, пулсном електрофорезом и Фемто Пулсном анализом.
Протокол (1) даје веома ниске приносе, било кад се користи самостално или као
вид пречишћавања ДНК изоловане на други начин. Протоколом (3) се добија висок
принос ДНК незадовољавајућег квалитета. Најквалитетнију ДНК у количинама
довољним за секвенцирање су дали протоколи (2) и (4).
PB  - Beograd : Srpsko biološko društvo
C3  - Treći kongres biologa Srbije
T1  - Usavršavanje protokola za izolaciju DNK visoke molekularne mase iz kičice (Centaurium erythraea)
T1  - Усавршавање протокола за изолацију ДНК високе молекуларне масе из кичице (Centaurium erythraea)
EP  - 295
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1743
ER  - 

@conference{
author = "Simonović, Ana and Filipović, Biljana and Banjanac, Tijana and Kojić, Milan and Ćuković, Katarina and Subotić, Angelina",
year = "2022",
abstract = "Biljka kičica (Centaurium erythraea) je interesantna ne samo zbog svojih
lekovitih svojstava, već i zbog izuzetne razvojne plastičnosti i regenerative
sposobnosti u in vitro kulturi. Kako bi identifikovali gene uključene u
morfogenetske procese, sekundarni metabolizam i odgovore na stres, pored
sekvenciranih transkriptoma1 je potreban i sekvenciran genom. Za sekvenciranje
genoma najmodernijom PacBio tehnologijom, neophodna je veoma čista
visokomolekularna DNK, čijih 90% čine lanci > 20 kb. DNK je izolovana iz
mladih listova diploidnog varijeteta kičice gajene in vitro, korišćenjem četiri
različita protokola: (1) Quick-DNA HMW MagBead Kit (Zymo Research, #D6060);
(2) modifikovani CTAB protokol;2 (3) protokol za izolaciju bakterijske DNK
prilagođen biljnim tkivima i (4) izmenjen protokol za izolaciju RNK iz bora.3
Kvalitet DNK izolata je proveravan spektrofotometrijski Nano Drop aparatom,
običnom elektroforezom, pulsnom elektroforezom i Femto Pulsnom analizom.
Protokol (1) daje veoma niske prinose, bilo kad se koristi samostalno ili kao
vid prečišćavanja DNK izolovane na drugi način. Protokolom (3) se dobija visok
prinos DNK nezadovoljavajućeg kvaliteta. Najkvalitetniju DNK u količinama
dovoljnim za sekvenciranje su dali protokoli (2) i (4)., Биљка кичица (Centaurium erythraea) је интересантна не само због својих
лековитих својстава, већ и због изузетне развојне пластичности и регенеративе
способности у in vitro култури. Како би идентификовали гене укључене у
морфогенетске процесе, секундарни метаболизам и одговоре на стрес, поред
секвенцираних транскриптома1 је потребан и секвенциран геном. За секвенцирање
генома најмодернијом PacBio технологијом, неопходна је веома чиста
високомолекуларна ДНК, чијих 90% чине ланци > 20 kb. ДНК је изолована из
младих листова диплоидног варијетета кичице гајене in vitro, коришћењем четири
различита протокола: (1) Quick-DNA HMW MagBead Kit (Zymo Research, #D6060);
(2) модификовани ЦТАБ протокол;2 (3) протокол за изолацију бактеријске ДНК
прилагођен биљним ткивима и (4) измењен протокол за изолацију РНК из бора.3
Квалитет ДНК изолата је провераван спектрофотометријски Nano Drop апаратом,
обичном електрофорезом, пулсном електрофорезом и Фемто Пулсном анализом.
Протокол (1) даје веома ниске приносе, било кад се користи самостално или као
вид пречишћавања ДНК изоловане на други начин. Протоколом (3) се добија висок
принос ДНК незадовољавајућег квалитета. Најквалитетнију ДНК у количинама
довољним за секвенцирање су дали протоколи (2) и (4).",
publisher = "Beograd : Srpsko biološko društvo",
journal = "Treći kongres biologa Srbije",
title = "Usavršavanje protokola za izolaciju DNK visoke molekularne mase iz kičice (Centaurium erythraea), Усавршавање протокола за изолацију ДНК високе молекуларне масе из кичице (Centaurium erythraea)",
pages = "295",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1743"
}

Simonović, A., Filipović, B., Banjanac, T., Kojić, M., Ćuković, K.,& Subotić, A.. (2022). Usavršavanje protokola za izolaciju DNK visoke molekularne mase iz kičice (Centaurium erythraea). in Treći kongres biologa Srbije
Beograd : Srpsko biološko društvo..
https://hdl.handle.net/21.15107/rcub_imagine_1743

Simonović A, Filipović B, Banjanac T, Kojić M, Ćuković K, Subotić A. Usavršavanje protokola za izolaciju DNK visoke molekularne mase iz kičice (Centaurium erythraea). in Treći kongres biologa Srbije. 2022;:null-295.
https://hdl.handle.net/21.15107/rcub_imagine_1743 .

Simonović, Ana, Filipović, Biljana, Banjanac, Tijana, Kojić, Milan, Ćuković, Katarina, Subotić, Angelina, "Usavršavanje protokola za izolaciju DNK visoke molekularne mase iz kičice (Centaurium erythraea)" in Treći kongres biologa Srbije (2022),
https://hdl.handle.net/21.15107/rcub_imagine_1743 .

TY  - CONF
AU  - Kojić, Milan
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1752
AB  - Nastanak i širenje antibiotske rezistencije predstavlja sve veći problem u
lečenju infektivnih bolesti, posebno kod osetljivih osoba. Problem u lečenju
već sada odnosi preko milion ljudskih života godišnje, a očekuje se da će taj broj
rasti i 2050 dostići 10 miliona, tako da rešenje ovog problema zahteva
angažovanost nauke, medicine i celokupnog društva. Takođe ovde treba dodati i
štete koje ovaj problem nanosi u veterinarskoj medicini i poljoprivredi. S
obzirom da je problem nastao usled prekomerne upotrebe i zloupotrebe
antibiotika jedno od rešenja jeste njihova ispravna upotreba. Pored toga postoji
potreba za otkrivanjem novih antimikrobnih agenasa koji bi se koristili kao
dopuna ili zamena za antibiotike kako se civilizacija ne bi vratila u
preantibiotsko doba. Perspektive za otkrivanje novih potentnih agenasa poput
antibiotika su male tako da su istraživanja fokusirana na veći broj alternativa
kao što su: antimikrobni peptidi (u kliničkoj upotrebi se već nalazi nekoliko
peptidnih antibiotika kao što su bacitracin i kolistin), različiti produkti
biljaka, hemijski sintetisana antimikrobna jedinjenja, specifična antitela,
vakcine, probiotici, bakteriofagi i drugi molekuli. U rešavanje ovog globalnog
problema naučnici su uključili i veštačku inteligenciju (mašinsko učenje)
kako bi na što brži način iskoristili dosadašnja saznanja za dizajniranje novih
jedinjenja i himernih molekula koja bi bila dostojna zamena kliničkim
antibioticima, što je već rezultovalo u otkrivanju novog veoma potentnog
antibakterijskog jedinjenja, halicina. Svesnost o ozbiljnosti problema u kome se
ceo svet nalazi se mora podići na viši nivo i to kroz angažovanje svih
relevantnih subjekata, a posebno nauke, medicine, službi javnog informisanja i
vlada svih država sveta.
AB  - Настанак и ширење антибиотске резистенције представља све већи проблем у
лечењу инфективних болести, посебно код осетљивих особа. Проблем у лечењу
већ сада односи преко милион људских живота годишње, а очекује се да ће тај број
расти и 2050 достићи 10 милиона, тако да решење овог проблема захтева
ангажованост науке, медицине и целокупног друштва. Такође овде треба додати и
штете које овај проблем наноси у ветеринарској медицини и пољопривреди. С
обзиром да је проблем настао услед прекомерне употребе и злоупотребе
антибиотика једно од решења јесте њихова исправна употреба. Поред тога постоји
потреба за откривањем нових антимикробних агенаса који би се користили као
допуна или замена за антибиотике како се цивилизација не би вратила у
преантибиотско доба. Перспективе за откривање нових потентних агенаса попут
антибиотика су мале тако да су истраживања фокусирана на већи број алтернатива
као што су: антимикробни пептиди (у клиничкој употреби се већ налази неколико
пептидних антибиотика као што су бацитрацин и колистин), различити продукти
биљака, хемијски синтетисана антимикробна једињења, специфична антитела,
вакцине, пробиотици, бактериофаги и други молекули. У решавање овог глобалног
проблема научници су укључили и вештачку интелигенцију (машинско учење)
како би на што бржи начин искористили досадашња сазнања за дизајнирање нових
једињења и химерних молекула која би била достојна замена клиничким
антибиотицима, што је већ резултовало у откривању новог веома потентног
антибактеријског једињења, халицина. Свесност о озбиљности проблема у коме се
цео свет налази се мора подићи на виши ниво и то кроз ангажовање свих
релевантних субјеката, а посебно науке, медицине, служби јавног информисања и
влада свих држава света.
PB  - Beograd : Srpsko biološko društvo
C3  - Treći kongres biologa Srbije
T1  - Perspektive za otkrivanje novih antimikrobnih agenasa za borbu protiv multi-rezistentnih patogena
T1  - Перспективе за откривање нових антимикробних агенаса за борбу против мулти-резистентних патогена
SP  - 248
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1752
ER  - 

@conference{
author = "Kojić, Milan",
year = "2022",
abstract = "Nastanak i širenje antibiotske rezistencije predstavlja sve veći problem u
lečenju infektivnih bolesti, posebno kod osetljivih osoba. Problem u lečenju
već sada odnosi preko milion ljudskih života godišnje, a očekuje se da će taj broj
rasti i 2050 dostići 10 miliona, tako da rešenje ovog problema zahteva
angažovanost nauke, medicine i celokupnog društva. Takođe ovde treba dodati i
štete koje ovaj problem nanosi u veterinarskoj medicini i poljoprivredi. S
obzirom da je problem nastao usled prekomerne upotrebe i zloupotrebe
antibiotika jedno od rešenja jeste njihova ispravna upotreba. Pored toga postoji
potreba za otkrivanjem novih antimikrobnih agenasa koji bi se koristili kao
dopuna ili zamena za antibiotike kako se civilizacija ne bi vratila u
preantibiotsko doba. Perspektive za otkrivanje novih potentnih agenasa poput
antibiotika su male tako da su istraživanja fokusirana na veći broj alternativa
kao što su: antimikrobni peptidi (u kliničkoj upotrebi se već nalazi nekoliko
peptidnih antibiotika kao što su bacitracin i kolistin), različiti produkti
biljaka, hemijski sintetisana antimikrobna jedinjenja, specifična antitela,
vakcine, probiotici, bakteriofagi i drugi molekuli. U rešavanje ovog globalnog
problema naučnici su uključili i veštačku inteligenciju (mašinsko učenje)
kako bi na što brži način iskoristili dosadašnja saznanja za dizajniranje novih
jedinjenja i himernih molekula koja bi bila dostojna zamena kliničkim
antibioticima, što je već rezultovalo u otkrivanju novog veoma potentnog
antibakterijskog jedinjenja, halicina. Svesnost o ozbiljnosti problema u kome se
ceo svet nalazi se mora podići na viši nivo i to kroz angažovanje svih
relevantnih subjekata, a posebno nauke, medicine, službi javnog informisanja i
vlada svih država sveta., Настанак и ширење антибиотске резистенције представља све већи проблем у
лечењу инфективних болести, посебно код осетљивих особа. Проблем у лечењу
већ сада односи преко милион људских живота годишње, а очекује се да ће тај број
расти и 2050 достићи 10 милиона, тако да решење овог проблема захтева
ангажованост науке, медицине и целокупног друштва. Такође овде треба додати и
штете које овај проблем наноси у ветеринарској медицини и пољопривреди. С
обзиром да је проблем настао услед прекомерне употребе и злоупотребе
антибиотика једно од решења јесте њихова исправна употреба. Поред тога постоји
потреба за откривањем нових антимикробних агенаса који би се користили као
допуна или замена за антибиотике како се цивилизација не би вратила у
преантибиотско доба. Перспективе за откривање нових потентних агенаса попут
антибиотика су мале тако да су истраживања фокусирана на већи број алтернатива
као што су: антимикробни пептиди (у клиничкој употреби се већ налази неколико
пептидних антибиотика као што су бацитрацин и колистин), различити продукти
биљака, хемијски синтетисана антимикробна једињења, специфична антитела,
вакцине, пробиотици, бактериофаги и други молекули. У решавање овог глобалног
проблема научници су укључили и вештачку интелигенцију (машинско учење)
како би на што бржи начин искористили досадашња сазнања за дизајнирање нових
једињења и химерних молекула која би била достојна замена клиничким
антибиотицима, што је већ резултовало у откривању новог веома потентног
антибактеријског једињења, халицина. Свесност о озбиљности проблема у коме се
цео свет налази се мора подићи на виши ниво и то кроз ангажовање свих
релевантних субјеката, а посебно науке, медицине, служби јавног информисања и
влада свих држава света.",
publisher = "Beograd : Srpsko biološko društvo",
journal = "Treći kongres biologa Srbije",
title = "Perspektive za otkrivanje novih antimikrobnih agenasa za borbu protiv multi-rezistentnih patogena, Перспективе за откривање нових антимикробних агенаса за борбу против мулти-резистентних патогена",
pages = "248",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1752"
}

Kojić, M.. (2022). Perspektive za otkrivanje novih antimikrobnih agenasa za borbu protiv multi-rezistentnih patogena. in Treći kongres biologa Srbije
Beograd : Srpsko biološko društvo., 248.
https://hdl.handle.net/21.15107/rcub_imagine_1752

Kojić M. Perspektive za otkrivanje novih antimikrobnih agenasa za borbu protiv multi-rezistentnih patogena. in Treći kongres biologa Srbije. 2022;:248.
https://hdl.handle.net/21.15107/rcub_imagine_1752 .

Kojić, Milan, "Perspektive za otkrivanje novih antimikrobnih agenasa za borbu protiv multi-rezistentnih patogena" in Treći kongres biologa Srbije (2022):248,
https://hdl.handle.net/21.15107/rcub_imagine_1752 .

TY  - JOUR
AU  - Novović, Katarina
AU  - Malešević, Milka
AU  - Dinić, Miroslav
AU  - Gardijan, Lazar
AU  - Kojić, Milan
AU  - Jovčić, Branko
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1525
AB  - Signal transduction systems are the key players of bacterial adaptation and survival. The orthodox two-component signal transduction systems perceive diverse environmental stimuli and their regulatory response leads to cellular changes. Although rarely described, the unorthodox three-component systems are also implemented in the regulation of major bacterial behavior such as the virulence of clinically relevant pathogen P. aeruginosa. Previously, we described a novel three-component system in P. capeferrum WCS358 (RclSAR) where the sensor kinase RclS stimulates the intI1 transcription in stationary growth phase. In this study, using rclS knock-out mutant, we identified RclSAR regulon in P. capeferrum WCS358. The RNA sequencing revealed that activity of RclSAR signal transduction system is growth phase dependent with more pronounced regulatory potential in early stages of growth. Transcriptional analysis emphasized the role of RclSAR in global regulation and indicated the involvement of this system in regulation of diverse cellular activities such as RNA binding and metabolic and biocontrol processes. Importantly, phenotypic comparison of WCS358 wild type and Delta rclS mutant showed that RclS sensor kinase contributes to modulation of antibiotic resistance, production of AHLs and siderophore as well as host cell adherence and cytotoxicity. Finally, we proposed the improved model of interplay between RclSAR, RpoS and LasIR regulatory systems in P. capeferrum WCS358.
PB  - MDPI, Basel
T2  - International Journal of Molecular Sciences
T1  - RclS Sensor Kinase Modulates Virulence of Pseudomonas capeferrum
IS  - 15
VL  - 23
DO  - 10.3390/ijms23158232
ER  - 

@article{
author = "Novović, Katarina and Malešević, Milka and Dinić, Miroslav and Gardijan, Lazar and Kojić, Milan and Jovčić, Branko",
year = "2022",
abstract = "Signal transduction systems are the key players of bacterial adaptation and survival. The orthodox two-component signal transduction systems perceive diverse environmental stimuli and their regulatory response leads to cellular changes. Although rarely described, the unorthodox three-component systems are also implemented in the regulation of major bacterial behavior such as the virulence of clinically relevant pathogen P. aeruginosa. Previously, we described a novel three-component system in P. capeferrum WCS358 (RclSAR) where the sensor kinase RclS stimulates the intI1 transcription in stationary growth phase. In this study, using rclS knock-out mutant, we identified RclSAR regulon in P. capeferrum WCS358. The RNA sequencing revealed that activity of RclSAR signal transduction system is growth phase dependent with more pronounced regulatory potential in early stages of growth. Transcriptional analysis emphasized the role of RclSAR in global regulation and indicated the involvement of this system in regulation of diverse cellular activities such as RNA binding and metabolic and biocontrol processes. Importantly, phenotypic comparison of WCS358 wild type and Delta rclS mutant showed that RclS sensor kinase contributes to modulation of antibiotic resistance, production of AHLs and siderophore as well as host cell adherence and cytotoxicity. Finally, we proposed the improved model of interplay between RclSAR, RpoS and LasIR regulatory systems in P. capeferrum WCS358.",
publisher = "MDPI, Basel",
journal = "International Journal of Molecular Sciences",
title = "RclS Sensor Kinase Modulates Virulence of Pseudomonas capeferrum",
number = "15",
volume = "23",
doi = "10.3390/ijms23158232"
}

Novović, K., Malešević, M., Dinić, M., Gardijan, L., Kojić, M.,& Jovčić, B.. (2022). RclS Sensor Kinase Modulates Virulence of Pseudomonas capeferrum. in International Journal of Molecular Sciences
MDPI, Basel., 23(15).
https://doi.org/10.3390/ijms23158232

Novović K, Malešević M, Dinić M, Gardijan L, Kojić M, Jovčić B. RclS Sensor Kinase Modulates Virulence of Pseudomonas capeferrum. in International Journal of Molecular Sciences. 2022;23(15).
doi:10.3390/ijms23158232 .

Novović, Katarina, Malešević, Milka, Dinić, Miroslav, Gardijan, Lazar, Kojić, Milan, Jovčić, Branko, "RclS Sensor Kinase Modulates Virulence of Pseudomonas capeferrum" in International Journal of Molecular Sciences, 23, no. 15 (2022),
https://doi.org/10.3390/ijms23158232 . .

TY  - JOUR
AU  - Rakonjac, B.
AU  - Lepšanović, Z.
AU  - Šuljagić, V.
AU  - Jovčić, Branko
AU  - Kojić, Milan
AU  - Larsen, A.R.
AU  - Urić, M.
AU  - Ćirković, I.
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1544
AB  - Epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) is continually changing. Frequency of genotypes typical for community-associated MRSA (CA-MRSA) is increasing in hospitals, as well as resistance to antimicrobial agents. Moreover, different clones predominate in different geographic regions, and temporal shifts occur in the predominant clonal type. The aim of this study was to estimate the prevalence of MRSA, CAMRSA and PVL-positive MRSA isolates from patients hospitalised in the Military Medical Academy (MMA) and from outpatients, and to perform genotyping of PVL-positive MRSA isolates. MRSA isolates were obtained by standard microbiological techniques. PVL-positive MRSA were detected by single PCR. Determination of SCCmec types in MRSA isolates was done using multiplex PCR and genotyping of PVL-positive MRSA by PFGE, MLST and spa typing. The prevalence of MRSA among S. aureus isolates from different clinical specimens was 43.4%. In outpatients the prevalence of MRSA was 3.2%. SCCmec types specific for CA-MRSA were found in 26% of MRSA isolates from hospitalised patients. In groups, hospitalised patients and outpatients, the prevalence of PVL-positive MRSA isolates was 4%, and all of them harboured SCCmec type V genetic element. PFGE revealed minor differences between four groups of PVL-positive MRSA isolates, but all of them belonged to ST152, and all except one were of the t355 spa type. High prevalence of MRSA and CAMRSA in MMA, especially the presence of PVL-positive CA-MRSA, represent a serious health threat for patients. Genotype t355/ST152/SCCmec V is the dominant MRSA clone among PVL-positive CA-MRSA.
PB  - Public Library of Science
T2  - PLoS One
T1  - Predominance of t355/ST152/SCCmec V clonal type among PVL-positive MRSA isolates in a tertiary care hospital in Belgrade, Serbia
IS  - 9 September
VL  - 17
DO  - 10.1371/journal.pone.0273474
ER  - 

@article{
author = "Rakonjac, B. and Lepšanović, Z. and Šuljagić, V. and Jovčić, Branko and Kojić, Milan and Larsen, A.R. and Urić, M. and Ćirković, I.",
year = "2022",
abstract = "Epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) is continually changing. Frequency of genotypes typical for community-associated MRSA (CA-MRSA) is increasing in hospitals, as well as resistance to antimicrobial agents. Moreover, different clones predominate in different geographic regions, and temporal shifts occur in the predominant clonal type. The aim of this study was to estimate the prevalence of MRSA, CAMRSA and PVL-positive MRSA isolates from patients hospitalised in the Military Medical Academy (MMA) and from outpatients, and to perform genotyping of PVL-positive MRSA isolates. MRSA isolates were obtained by standard microbiological techniques. PVL-positive MRSA were detected by single PCR. Determination of SCCmec types in MRSA isolates was done using multiplex PCR and genotyping of PVL-positive MRSA by PFGE, MLST and spa typing. The prevalence of MRSA among S. aureus isolates from different clinical specimens was 43.4%. In outpatients the prevalence of MRSA was 3.2%. SCCmec types specific for CA-MRSA were found in 26% of MRSA isolates from hospitalised patients. In groups, hospitalised patients and outpatients, the prevalence of PVL-positive MRSA isolates was 4%, and all of them harboured SCCmec type V genetic element. PFGE revealed minor differences between four groups of PVL-positive MRSA isolates, but all of them belonged to ST152, and all except one were of the t355 spa type. High prevalence of MRSA and CAMRSA in MMA, especially the presence of PVL-positive CA-MRSA, represent a serious health threat for patients. Genotype t355/ST152/SCCmec V is the dominant MRSA clone among PVL-positive CA-MRSA.",
publisher = "Public Library of Science",
journal = "PLoS One",
title = "Predominance of t355/ST152/SCCmec V clonal type among PVL-positive MRSA isolates in a tertiary care hospital in Belgrade, Serbia",
number = "9 September",
volume = "17",
doi = "10.1371/journal.pone.0273474"
}

Rakonjac, B., Lepšanović, Z., Šuljagić, V., Jovčić, B., Kojić, M., Larsen, A.R., Urić, M.,& Ćirković, I.. (2022). Predominance of t355/ST152/SCCmec V clonal type among PVL-positive MRSA isolates in a tertiary care hospital in Belgrade, Serbia. in PLoS One
Public Library of Science., 17(9 September).
https://doi.org/10.1371/journal.pone.0273474

Rakonjac B, Lepšanović Z, Šuljagić V, Jovčić B, Kojić M, Larsen A, Urić M, Ćirković I. Predominance of t355/ST152/SCCmec V clonal type among PVL-positive MRSA isolates in a tertiary care hospital in Belgrade, Serbia. in PLoS One. 2022;17(9 September).
doi:10.1371/journal.pone.0273474 .

Rakonjac, B., Lepšanović, Z., Šuljagić, V., Jovčić, Branko, Kojić, Milan, Larsen, A.R., Urić, M., Ćirković, I., "Predominance of t355/ST152/SCCmec V clonal type among PVL-positive MRSA isolates in a tertiary care hospital in Belgrade, Serbia" in PLoS One, 17, no. 9 September (2022),
https://doi.org/10.1371/journal.pone.0273474 . .

TY  - JOUR
AU  - Novović, Katarina
AU  - Malešević, Milka
AU  - Gardijan, Lazar
AU  - Kojić, Milan
AU  - Jovčić, Branko
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1572
AB  - The rapid and appropriate response of Pseudomonas spp. to environmental fluctuations has been enabled by numerous signal transduction regulatory systems. Regulatory systems in Pseudomonas aeruginosa are organized in a complex network which provides quick and fine-tuned cellular response through regulation of virulence and antibiotic resistance determinants production. Mobile integrons represent genetic elements included in the rapid dissemination of multiple antibiotic resistance determinants. The key factor of integron dynamics is enzyme integrase. So far, global regulators LexA, RpoS and PsrA have been recognized as regulators of the intI1 transcription. In this study, we discovered novel activator of the intI1 transcription, sensor kinase RclS, in Pseudomonas putida WCS358. This regulation is limited to stationary growth phase and appears to be indirect, at least through regulation of the rpoS expression. Sensor kinase RclS is a part of novel three-component system Rcl (Roc-like) together with two response regulators, RclR and RclA. RclS acted as a negative regulator of the rclA transcription, while the role in the rclR transcription regulation could not be defined. The RclSAR regulatory system seems to be a part of complex intI1 regulatory network which includes major stress response (SOS and RpoS) regulons.
PB  - Elsevier, Amsterdam
T2  - Research in Microbiology
T1  - Novel RclSAR three-component system regulates expression of the intI1 gene in the stationary growth phase
IS  - 1-2
VL  - 173
DO  - 10.1016/j.resmic.2021.103885
ER  - 

@article{
author = "Novović, Katarina and Malešević, Milka and Gardijan, Lazar and Kojić, Milan and Jovčić, Branko",
year = "2022",
abstract = "The rapid and appropriate response of Pseudomonas spp. to environmental fluctuations has been enabled by numerous signal transduction regulatory systems. Regulatory systems in Pseudomonas aeruginosa are organized in a complex network which provides quick and fine-tuned cellular response through regulation of virulence and antibiotic resistance determinants production. Mobile integrons represent genetic elements included in the rapid dissemination of multiple antibiotic resistance determinants. The key factor of integron dynamics is enzyme integrase. So far, global regulators LexA, RpoS and PsrA have been recognized as regulators of the intI1 transcription. In this study, we discovered novel activator of the intI1 transcription, sensor kinase RclS, in Pseudomonas putida WCS358. This regulation is limited to stationary growth phase and appears to be indirect, at least through regulation of the rpoS expression. Sensor kinase RclS is a part of novel three-component system Rcl (Roc-like) together with two response regulators, RclR and RclA. RclS acted as a negative regulator of the rclA transcription, while the role in the rclR transcription regulation could not be defined. The RclSAR regulatory system seems to be a part of complex intI1 regulatory network which includes major stress response (SOS and RpoS) regulons.",
publisher = "Elsevier, Amsterdam",
journal = "Research in Microbiology",
title = "Novel RclSAR three-component system regulates expression of the intI1 gene in the stationary growth phase",
number = "1-2",
volume = "173",
doi = "10.1016/j.resmic.2021.103885"
}

Novović, K., Malešević, M., Gardijan, L., Kojić, M.,& Jovčić, B.. (2022). Novel RclSAR three-component system regulates expression of the intI1 gene in the stationary growth phase. in Research in Microbiology
Elsevier, Amsterdam., 173(1-2).
https://doi.org/10.1016/j.resmic.2021.103885

Novović K, Malešević M, Gardijan L, Kojić M, Jovčić B. Novel RclSAR three-component system regulates expression of the intI1 gene in the stationary growth phase. in Research in Microbiology. 2022;173(1-2).
doi:10.1016/j.resmic.2021.103885 .

Novović, Katarina, Malešević, Milka, Gardijan, Lazar, Kojić, Milan, Jovčić, Branko, "Novel RclSAR three-component system regulates expression of the intI1 gene in the stationary growth phase" in Research in Microbiology, 173, no. 1-2 (2022),
https://doi.org/10.1016/j.resmic.2021.103885 . .

TY  - JOUR
AU  - Filipić, Brankica
AU  - Malešević, Milka
AU  - Vasiljević, Zorica
AU  - Novović, Katarina
AU  - Kojić, Milan
AU  - Jovčić, Branko
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1778
AB  - Trimethoprim-sulfamethoxazole (SXT) is the preferable treatment option of the infections caused by Achromobacter spp. Our study aimed to analyze the SXT resistance of 98 Achromobacter spp. isolates from pediatric patients, among which 33 isolates were SXT-resistant. The presence of intI1 was screened by PCR and genome sequence analyses. The intI1 gene was detected in 10 of SXT-resistant isolates that had shorter intI1 PCR fragments named intI1S. Structural changes in intI1S were confirmed by genome sequencing and analyses which revealed 86 amino acids deletion in IntI1S protein compared to canonical IntI1 protein. All IntI1S isolates were of non-CF origin. Pan-genome analysis of intI1S bearing A. xylosoxidans isolates comprised 9052 genes, with the core genome consisting of 5455 protein-coding genes. Results in this study indicate that IntI1S isolates were derived from clinical settings and that cystic fibrosis (CF) patients were potential reservoirs for healthcare-associated infections that occurred in non-CF patients.
PB  - Springer Science and Business Media B.V.
T2  - Folia Microbiologica
T1  - Comparative genomics of trimethoprim-sulfamethoxazole-resistant Achromobacter xylosoxidans clinical isolates from Serbia reveals shortened variant of class 1 integron integrase gene
DO  - 10.1007/s12223-022-01026-8
ER  - 

@article{
author = "Filipić, Brankica and Malešević, Milka and Vasiljević, Zorica and Novović, Katarina and Kojić, Milan and Jovčić, Branko",
year = "2022",
abstract = "Trimethoprim-sulfamethoxazole (SXT) is the preferable treatment option of the infections caused by Achromobacter spp. Our study aimed to analyze the SXT resistance of 98 Achromobacter spp. isolates from pediatric patients, among which 33 isolates were SXT-resistant. The presence of intI1 was screened by PCR and genome sequence analyses. The intI1 gene was detected in 10 of SXT-resistant isolates that had shorter intI1 PCR fragments named intI1S. Structural changes in intI1S were confirmed by genome sequencing and analyses which revealed 86 amino acids deletion in IntI1S protein compared to canonical IntI1 protein. All IntI1S isolates were of non-CF origin. Pan-genome analysis of intI1S bearing A. xylosoxidans isolates comprised 9052 genes, with the core genome consisting of 5455 protein-coding genes. Results in this study indicate that IntI1S isolates were derived from clinical settings and that cystic fibrosis (CF) patients were potential reservoirs for healthcare-associated infections that occurred in non-CF patients.",
publisher = "Springer Science and Business Media B.V.",
journal = "Folia Microbiologica",
title = "Comparative genomics of trimethoprim-sulfamethoxazole-resistant Achromobacter xylosoxidans clinical isolates from Serbia reveals shortened variant of class 1 integron integrase gene",
doi = "10.1007/s12223-022-01026-8"
}

Filipić, B., Malešević, M., Vasiljević, Z., Novović, K., Kojić, M.,& Jovčić, B.. (2022). Comparative genomics of trimethoprim-sulfamethoxazole-resistant Achromobacter xylosoxidans clinical isolates from Serbia reveals shortened variant of class 1 integron integrase gene. in Folia Microbiologica
Springer Science and Business Media B.V...
https://doi.org/10.1007/s12223-022-01026-8

Filipić B, Malešević M, Vasiljević Z, Novović K, Kojić M, Jovčić B. Comparative genomics of trimethoprim-sulfamethoxazole-resistant Achromobacter xylosoxidans clinical isolates from Serbia reveals shortened variant of class 1 integron integrase gene. in Folia Microbiologica. 2022;.
doi:10.1007/s12223-022-01026-8 .

Filipić, Brankica, Malešević, Milka, Vasiljević, Zorica, Novović, Katarina, Kojić, Milan, Jovčić, Branko, "Comparative genomics of trimethoprim-sulfamethoxazole-resistant Achromobacter xylosoxidans clinical isolates from Serbia reveals shortened variant of class 1 integron integrase gene" in Folia Microbiologica (2022),
https://doi.org/10.1007/s12223-022-01026-8 . .

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  - Jovčić, Branko
AU  - Malešević, Milka
AU  - Kojić, Milan
AU  - Galić, Nataša
AU  - Todorović, Dalibor
AU  - Vidanović, Dejan
AU  - Velhner, Maja
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1578
AB  - Owing to the emerging resistance to antimicrobials in Salmonella Kentucky isolates around the globe, the genomic comparison of all the registered multidrug-resistant Salmonella Kentucky isolates in Serbia (five from humans, one from turkey flock, and one from meat) was done. Most of the isolates were isolated from patients returning from Egypt or Tunisia or originated from imported turkey flock and turkey meat. The comparative analysis of resistance and virulence genes was done. All isolates belonged to sequence type-ST198 and were resistant to ciprofloxacin (Cip). The resistance to Cip was mediated by target mutations of the gyrA and parC genes, which encode topoisomerase I and II, respectively. Multidrug-resistant phenotype to aminoglycosides, beta-lactam antibiotics, sulfonamides, and tetracyclines was detected in five isolates. However, none of the isolates was pan-resistant to antimicrobials. The number of single nucleotide polymorphisms between isolates varied from 8 to 43 and phylogenomics revealed the genetic proximity of the human isolate 10475/11 and the turkey meat isolate 5264/14, indicating a possible meat-to-human transfer. All isolates belonged to the main Salmonella Kentucky MDR lineage, carrying the Salmonella genomic island 1 (SGI1-K) subtype. The SGI1-K of Serbian isolates showed mosaicism attributed to rapid intraclonal evolution. Many virulence factors were detected in all the isolates, including SPI-1, SPI-2, SPI-3, SPI-4, SPI-5, SPI-9, and C63PI. Although Salmonella Kentucky has rarely been isolated from humans, food, and animals in Serbia, further surveillance is needed to diminish the risk of the spreading of resistant clones and their meat-to-human transmission.
PB  - Mary Ann Liebert, Inc, New Rochelle
T2  - Foodborne Pathogens and Disease
T1  - Genomic Analysis of Multidrug-Resistant Salmonella enterica Serovar Kentucky Isolates from Humans, Turkey, and Food in the Republic of Serbia
EP  - 636
IS  - 9
SP  - 630
VL  - 19
DO  - 10.1089/fpd.2022.0029
ER  - 

@article{
author = "Jovčić, Branko and Malešević, Milka and Kojić, Milan and Galić, Nataša and Todorović, Dalibor and Vidanović, Dejan and Velhner, Maja",
year = "2022",
abstract = "Owing to the emerging resistance to antimicrobials in Salmonella Kentucky isolates around the globe, the genomic comparison of all the registered multidrug-resistant Salmonella Kentucky isolates in Serbia (five from humans, one from turkey flock, and one from meat) was done. Most of the isolates were isolated from patients returning from Egypt or Tunisia or originated from imported turkey flock and turkey meat. The comparative analysis of resistance and virulence genes was done. All isolates belonged to sequence type-ST198 and were resistant to ciprofloxacin (Cip). The resistance to Cip was mediated by target mutations of the gyrA and parC genes, which encode topoisomerase I and II, respectively. Multidrug-resistant phenotype to aminoglycosides, beta-lactam antibiotics, sulfonamides, and tetracyclines was detected in five isolates. However, none of the isolates was pan-resistant to antimicrobials. The number of single nucleotide polymorphisms between isolates varied from 8 to 43 and phylogenomics revealed the genetic proximity of the human isolate 10475/11 and the turkey meat isolate 5264/14, indicating a possible meat-to-human transfer. All isolates belonged to the main Salmonella Kentucky MDR lineage, carrying the Salmonella genomic island 1 (SGI1-K) subtype. The SGI1-K of Serbian isolates showed mosaicism attributed to rapid intraclonal evolution. Many virulence factors were detected in all the isolates, including SPI-1, SPI-2, SPI-3, SPI-4, SPI-5, SPI-9, and C63PI. Although Salmonella Kentucky has rarely been isolated from humans, food, and animals in Serbia, further surveillance is needed to diminish the risk of the spreading of resistant clones and their meat-to-human transmission.",
publisher = "Mary Ann Liebert, Inc, New Rochelle",
journal = "Foodborne Pathogens and Disease",
title = "Genomic Analysis of Multidrug-Resistant Salmonella enterica Serovar Kentucky Isolates from Humans, Turkey, and Food in the Republic of Serbia",
pages = "636-630",
number = "9",
volume = "19",
doi = "10.1089/fpd.2022.0029"
}

Jovčić, B., Malešević, M., Kojić, M., Galić, N., Todorović, D., Vidanović, D.,& Velhner, M.. (2022). Genomic Analysis of Multidrug-Resistant Salmonella enterica Serovar Kentucky Isolates from Humans, Turkey, and Food in the Republic of Serbia. in Foodborne Pathogens and Disease
Mary Ann Liebert, Inc, New Rochelle., 19(9), 630-636.
https://doi.org/10.1089/fpd.2022.0029

Jovčić B, Malešević M, Kojić M, Galić N, Todorović D, Vidanović D, Velhner M. Genomic Analysis of Multidrug-Resistant Salmonella enterica Serovar Kentucky Isolates from Humans, Turkey, and Food in the Republic of Serbia. in Foodborne Pathogens and Disease. 2022;19(9):630-636.
doi:10.1089/fpd.2022.0029 .

Jovčić, Branko, Malešević, Milka, Kojić, Milan, Galić, Nataša, Todorović, Dalibor, Vidanović, Dejan, Velhner, Maja, "Genomic Analysis of Multidrug-Resistant Salmonella enterica Serovar Kentucky Isolates from Humans, Turkey, and Food in the Republic of Serbia" in Foodborne Pathogens and Disease, 19, no. 9 (2022):630-636,
https://doi.org/10.1089/fpd.2022.0029 . .

TY  - JOUR
AU  - Ušjak, Dušan
AU  - Novović, Katarina
AU  - Filipić, Brankica
AU  - Kojić, Milan
AU  - Filipović, Nenad
AU  - Stevanović, Magdalena
AU  - Milenković, Marina T.
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1600
AB  - Aims To investigate the synergistic activity of colistin and selenium nanoparticles (SeNPs) against pandrug-resistant (PDR) Ac. baumannii. Methods and Results Chequerboard and time-kill assays were employed to explore the potential synergistic interactions between colistin and SeNPs against Ac. baumannii isolates (8), previously determined as colistin-resistant (MIC range 16-256 mu g ml(-1)). Also, whole-genome sequencing (WGS) and gene expression analyses were used to elucidate the mechanisms of colistin resistance. Exceptionally strong synergistic activity (FICI range 0.004-0.035) of colistin and SeNPs against colistin-resistant isolates was revealed. Colistin (0.5 or 1 mu g ml(-1)) used in combination with SeNPs (0.5 mu g ml(-1)) was able to reduce initial inoculum during the first 4 h of incubation, in contrast to colistin (0.5, 1 or 2 mu g ml(-1)) alone. Conclusions These findings propose colistin/SeNPs combination as a new option to fight PDR Ac. baumannii, the therapeutic possibilities of which should be proved in future in vivo studies. Significance and Impact of Study Here we present the first evidence of synergy between colistin and selenium compounds against bacteria in general. Also, WGS and gene expression analyses provide some new insights into Ac. baumannii colistin resistance mechanisms.
PB  - Wiley, Hoboken
T2  - Journal of Applied Microbiology
T1  - In vitro colistin susceptibility of pandrug-resistant Ac. baumannii is restored in the presence of selenium nanoparticles
EP  - 1206
IS  - 3
SP  - 1197
VL  - 133
DO  - 10.1111/jam.15638
ER  - 

@article{
author = "Ušjak, Dušan and Novović, Katarina and Filipić, Brankica and Kojić, Milan and Filipović, Nenad and Stevanović, Magdalena and Milenković, Marina T.",
year = "2022",
abstract = "Aims To investigate the synergistic activity of colistin and selenium nanoparticles (SeNPs) against pandrug-resistant (PDR) Ac. baumannii. Methods and Results Chequerboard and time-kill assays were employed to explore the potential synergistic interactions between colistin and SeNPs against Ac. baumannii isolates (8), previously determined as colistin-resistant (MIC range 16-256 mu g ml(-1)). Also, whole-genome sequencing (WGS) and gene expression analyses were used to elucidate the mechanisms of colistin resistance. Exceptionally strong synergistic activity (FICI range 0.004-0.035) of colistin and SeNPs against colistin-resistant isolates was revealed. Colistin (0.5 or 1 mu g ml(-1)) used in combination with SeNPs (0.5 mu g ml(-1)) was able to reduce initial inoculum during the first 4 h of incubation, in contrast to colistin (0.5, 1 or 2 mu g ml(-1)) alone. Conclusions These findings propose colistin/SeNPs combination as a new option to fight PDR Ac. baumannii, the therapeutic possibilities of which should be proved in future in vivo studies. Significance and Impact of Study Here we present the first evidence of synergy between colistin and selenium compounds against bacteria in general. Also, WGS and gene expression analyses provide some new insights into Ac. baumannii colistin resistance mechanisms.",
publisher = "Wiley, Hoboken",
journal = "Journal of Applied Microbiology",
title = "In vitro colistin susceptibility of pandrug-resistant Ac. baumannii is restored in the presence of selenium nanoparticles",
pages = "1206-1197",
number = "3",
volume = "133",
doi = "10.1111/jam.15638"
}

Ušjak, D., Novović, K., Filipić, B., Kojić, M., Filipović, N., Stevanović, M.,& Milenković, M. T.. (2022). In vitro colistin susceptibility of pandrug-resistant Ac. baumannii is restored in the presence of selenium nanoparticles. in Journal of Applied Microbiology
Wiley, Hoboken., 133(3), 1197-1206.
https://doi.org/10.1111/jam.15638

Ušjak D, Novović K, Filipić B, Kojić M, Filipović N, Stevanović M, Milenković MT. In vitro colistin susceptibility of pandrug-resistant Ac. baumannii is restored in the presence of selenium nanoparticles. in Journal of Applied Microbiology. 2022;133(3):1197-1206.
doi:10.1111/jam.15638 .

Ušjak, Dušan, Novović, Katarina, Filipić, Brankica, Kojić, Milan, Filipović, Nenad, Stevanović, Magdalena, Milenković, Marina T., "In vitro colistin susceptibility of pandrug-resistant Ac. baumannii is restored in the presence of selenium nanoparticles" in Journal of Applied Microbiology, 133, no. 3 (2022):1197-1206,
https://doi.org/10.1111/jam.15638 . .

TY  - CONF
AU  - Malešević, Milka
AU  - Rašić, Slađan
AU  - Santra, Violeta
AU  - Kojić, Milan
AU  - Stanisavljević, Nemanja
PY  - 2021
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1871
AB  - Among them, bacterial and fungal pathogens Paenibacillus larvae, Melissococcus pluton, Ascosphera apis and
Nosema ceranae play a major impact on honey bees colonies. Thus, developing alternative prophylactic and
curative strategies are urgently needed. The use of probiotic bacteria in honey bee supplemental feeding is
therefore promising to treat or prevent diseases. Brevibacillus laterosporus, Gram-positive endospore forming
bacilli, is recognised as one of the promising antibacterial and antifungal agents producer.
The aim of this study was to examine the short-therm effects of B. laterosporus supplemented diet on worker
honey bee microbiome.
Dry spores of B. laterosporus strain BGSP11 have been administrated through a sugar syrup diet to ten
colonies and a representative specimen of worker honey bees was taken before the start of the treatment
and immediately after the syrup was consumed. The microbial diversity was assessed before and after the
treatment using Illumina MiSeq sequencing platforms (ID Genomics service, Seattle, WA, USA). 16s rRNA
sequencing for bacterial community profiling and fungal Internally Transcribes Spacer for mycological taxa
profiling were used. The next-generation microbiome bioinformatics platform QIIME2 v 2021.4 was used for
filtering and denoising obtained sequences, calculation of diversity metrics and taxonomy assignment. The
feature classifier was trained using the Greengenes v 13_8 for bacterial taxa and fungal UNITE database v 8.3.
The results obtained in this study indicated statisticaly significant alfa diversity between control and experimental
group honey bee microbiota composition. The diversity abundance was higher in control comparing
to the group treated with B. laterosporus strain BGSP11 spores. There was no significant diference in Bray-
Curtis distance among two groups of analysed samples. Regarding to mycological abundance, composition
was completely different between two groups; control group had Claviceps as predominant genus, while in
treated group of honey bee microbiome Metschnikowia genus was prevalent, indicating that the presence of
fungal pathogens in treated group is highly diminished.
PB  - Novi Sad : Faculty of Sciences, Department of Biology and Ecology
C3  - Biologia Serbica
T1  - Brevibacillus laterosporus supplementation diet modulates honey bee microbiome
IS  - 1 (Special Edition)
SP  - 113
VL  - 43
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1871
ER  - 

@conference{
author = "Malešević, Milka and Rašić, Slađan and Santra, Violeta and Kojić, Milan and Stanisavljević, Nemanja",
year = "2021",
abstract = "Among them, bacterial and fungal pathogens Paenibacillus larvae, Melissococcus pluton, Ascosphera apis and
Nosema ceranae play a major impact on honey bees colonies. Thus, developing alternative prophylactic and
curative strategies are urgently needed. The use of probiotic bacteria in honey bee supplemental feeding is
therefore promising to treat or prevent diseases. Brevibacillus laterosporus, Gram-positive endospore forming
bacilli, is recognised as one of the promising antibacterial and antifungal agents producer.
The aim of this study was to examine the short-therm effects of B. laterosporus supplemented diet on worker
honey bee microbiome.
Dry spores of B. laterosporus strain BGSP11 have been administrated through a sugar syrup diet to ten
colonies and a representative specimen of worker honey bees was taken before the start of the treatment
and immediately after the syrup was consumed. The microbial diversity was assessed before and after the
treatment using Illumina MiSeq sequencing platforms (ID Genomics service, Seattle, WA, USA). 16s rRNA
sequencing for bacterial community profiling and fungal Internally Transcribes Spacer for mycological taxa
profiling were used. The next-generation microbiome bioinformatics platform QIIME2 v 2021.4 was used for
filtering and denoising obtained sequences, calculation of diversity metrics and taxonomy assignment. The
feature classifier was trained using the Greengenes v 13_8 for bacterial taxa and fungal UNITE database v 8.3.
The results obtained in this study indicated statisticaly significant alfa diversity between control and experimental
group honey bee microbiota composition. The diversity abundance was higher in control comparing
to the group treated with B. laterosporus strain BGSP11 spores. There was no significant diference in Bray-
Curtis distance among two groups of analysed samples. Regarding to mycological abundance, composition
was completely different between two groups; control group had Claviceps as predominant genus, while in
treated group of honey bee microbiome Metschnikowia genus was prevalent, indicating that the presence of
fungal pathogens in treated group is highly diminished.",
publisher = "Novi Sad : Faculty of Sciences, Department of Biology and Ecology",
journal = "Biologia Serbica",
title = "Brevibacillus laterosporus supplementation diet modulates honey bee microbiome",
number = "1 (Special Edition)",
pages = "113",
volume = "43",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1871"
}

Malešević, M., Rašić, S., Santra, V., Kojić, M.,& Stanisavljević, N.. (2021). Brevibacillus laterosporus supplementation diet modulates honey bee microbiome. in Biologia Serbica
Novi Sad : Faculty of Sciences, Department of Biology and Ecology., 43(1 (Special Edition)), 113.
https://hdl.handle.net/21.15107/rcub_imagine_1871

Malešević M, Rašić S, Santra V, Kojić M, Stanisavljević N. Brevibacillus laterosporus supplementation diet modulates honey bee microbiome. in Biologia Serbica. 2021;43(1 (Special Edition)):113.
https://hdl.handle.net/21.15107/rcub_imagine_1871 .

Malešević, Milka, Rašić, Slađan, Santra, Violeta, Kojić, Milan, Stanisavljević, Nemanja, "Brevibacillus laterosporus supplementation diet modulates honey bee microbiome" in Biologia Serbica, 43, no. 1 (Special Edition) (2021):113,
https://hdl.handle.net/21.15107/rcub_imagine_1871 .

TY  - JOUR
AU  - Malešević, Milka
AU  - Stanisavljević, Nemanja
AU  - Miljković, Marija
AU  - Jovčić, Branko
AU  - Filipić, Brankica
AU  - Studholme, David J.
AU  - Kojić, Milan
PY  - 2021
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1507
AB  - Plasmids are autonomous episomally replicating genetic elements, which carry backbone genes important for the replication and maintenance within their host, and accessory genes that might confer an advantage to their host under specific selective pressure in its ecological niche. The genome of dairy isolate L. lactis subsp. lactis by. diacetylactis S50 was sequenced using the PacBio SMRT Cell Seq-RSII platform and revealed to possess one of the largest plasmidomes among L. lactis strains studied so far, harboring six plasmids: pS6 (5553 bp), pS7a (7308 bp), pS7b (7266 bp), pS19 (19,027 bp), pS74 (74,256 bp) and pS127 (127,002 bp) in total representing 8.9% of genome size (240,412 bp). Based on predicted plasmid replication proteins and origins it appears that all six plasmids replicate via the theta-type mechanism. The two the largest plasmids (pS74 and pS127), carry a number of genes known to be important for growth and survival in the dairy environment. These genes encode technological functions such as bacteriocin production, protein degradation, magnesium and cobalt/nickel transporters, selenium binding, exopolysaccharides (EPS) production, bacteriophage and stress resistance. Beside genes for replication, the small plasmids (pS6, pS7a, pS7a, and pS19) also carry genes important for mobilization and host survival such as type I restriction-modification (R-M) system, metal transporters, enzymes and transcriptional regulators. All plasmids in S50 strain are mobilizable, containing an oriT sequences, while pS127 is self-conjugative and allows for mobilization of the other plasmids. Small plasmids are prone to structural and segregational instability, while pS127 appeared to be segregationally stable thanks to the possession of two partition systems. The main characteristic of plasmid p574 is EPS production, while plasmid p5127 is characterized by proteinase and multiple bacteriocins, tra locus, phage abortive systems and metal transporters. In addition to LcnA and LcnB, plasmid p5127 encodes several bacteriocin-pheromone molecules and a new bacteriocin named LcnS50, with narrow spectrum of action limited to lactococci, that has been successfully cloned and heterologously expressed.
PB  - Elsevier, Amsterdam
T2  - International Journal of Food Microbiology
T1  - The large plasmidome of Lactococcus lactis subsp. lactis by. diacetylactis S50 confers its biotechnological properties
VL  - 337
DO  - 10.1016/j.ijfoodmicro.2020.108935
ER  - 

@article{
author = "Malešević, Milka and Stanisavljević, Nemanja and Miljković, Marija and Jovčić, Branko and Filipić, Brankica and Studholme, David J. and Kojić, Milan",
year = "2021",
abstract = "Plasmids are autonomous episomally replicating genetic elements, which carry backbone genes important for the replication and maintenance within their host, and accessory genes that might confer an advantage to their host under specific selective pressure in its ecological niche. The genome of dairy isolate L. lactis subsp. lactis by. diacetylactis S50 was sequenced using the PacBio SMRT Cell Seq-RSII platform and revealed to possess one of the largest plasmidomes among L. lactis strains studied so far, harboring six plasmids: pS6 (5553 bp), pS7a (7308 bp), pS7b (7266 bp), pS19 (19,027 bp), pS74 (74,256 bp) and pS127 (127,002 bp) in total representing 8.9% of genome size (240,412 bp). Based on predicted plasmid replication proteins and origins it appears that all six plasmids replicate via the theta-type mechanism. The two the largest plasmids (pS74 and pS127), carry a number of genes known to be important for growth and survival in the dairy environment. These genes encode technological functions such as bacteriocin production, protein degradation, magnesium and cobalt/nickel transporters, selenium binding, exopolysaccharides (EPS) production, bacteriophage and stress resistance. Beside genes for replication, the small plasmids (pS6, pS7a, pS7a, and pS19) also carry genes important for mobilization and host survival such as type I restriction-modification (R-M) system, metal transporters, enzymes and transcriptional regulators. All plasmids in S50 strain are mobilizable, containing an oriT sequences, while pS127 is self-conjugative and allows for mobilization of the other plasmids. Small plasmids are prone to structural and segregational instability, while pS127 appeared to be segregationally stable thanks to the possession of two partition systems. The main characteristic of plasmid p574 is EPS production, while plasmid p5127 is characterized by proteinase and multiple bacteriocins, tra locus, phage abortive systems and metal transporters. In addition to LcnA and LcnB, plasmid p5127 encodes several bacteriocin-pheromone molecules and a new bacteriocin named LcnS50, with narrow spectrum of action limited to lactococci, that has been successfully cloned and heterologously expressed.",
publisher = "Elsevier, Amsterdam",
journal = "International Journal of Food Microbiology",
title = "The large plasmidome of Lactococcus lactis subsp. lactis by. diacetylactis S50 confers its biotechnological properties",
volume = "337",
doi = "10.1016/j.ijfoodmicro.2020.108935"
}

Malešević, M., Stanisavljević, N., Miljković, M., Jovčić, B., Filipić, B., Studholme, D. J.,& Kojić, M.. (2021). The large plasmidome of Lactococcus lactis subsp. lactis by. diacetylactis S50 confers its biotechnological properties. in International Journal of Food Microbiology
Elsevier, Amsterdam., 337.
https://doi.org/10.1016/j.ijfoodmicro.2020.108935

Malešević M, Stanisavljević N, Miljković M, Jovčić B, Filipić B, Studholme DJ, Kojić M. The large plasmidome of Lactococcus lactis subsp. lactis by. diacetylactis S50 confers its biotechnological properties. in International Journal of Food Microbiology. 2021;337.
doi:10.1016/j.ijfoodmicro.2020.108935 .

Malešević, Milka, Stanisavljević, Nemanja, Miljković, Marija, Jovčić, Branko, Filipić, Brankica, Studholme, David J., Kojić, Milan, "The large plasmidome of Lactococcus lactis subsp. lactis by. diacetylactis S50 confers its biotechnological properties" in International Journal of Food Microbiology, 337 (2021),
https://doi.org/10.1016/j.ijfoodmicro.2020.108935 . .