Rašić, Slađan

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Short-term effect of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome

Malešević, Milka; Stanisavljević, Nemanja; Rašić, Slađan; Vukotić, Goran; Gardijan, Lazar; Obradović, Mina; Kojić, Milan

(Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade, 2023)

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 .

Short-term effects of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome: a pilot study

Stanisavljević, Nemanja; Malešević, Milka; Rašić, Slađan; Vukotić, Goran; Gardijan, Lazar; Obradović, Mina; Kojić, Milan

(2023)

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 . .

Brevibacillus laterosporus supplementation diet modulates honey bee microbiome

Malešević, Milka; Rašić, Slađan; Santra, Violeta; Kojić, Milan; Stanisavljević, Nemanja

(Novi Sad : Faculty of Sciences, Department of Biology and Ecology, 2021)

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 .