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

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

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

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

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