Lactobacillus reuteri and Streptococcus oligofermentans are efficient colonizers in a multi-species model of cariogenic biofilm

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Lactobacillus reuteri and Streptococcus oligofermentans are efficient colonizers in a multi-species model of cariogenic biofilm. / Rikvold, Pernille; Grønkjær, Lene; Chen, Zhihui et al.

2021. Abstract from 68th ORCA Congress, Zagreb, Zagreb, Croatia.

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Rikvold, P, Grønkjær, L, Chen, Z, Göstemeyer, G, Schwendicke, F & Schlafer, S 2021, 'Lactobacillus reuteri and Streptococcus oligofermentans are efficient colonizers in a multi-species model of cariogenic biofilm', 68th ORCA Congress, Zagreb, Zagreb, Croatia, 07/07/2021 - 10/07/2021.

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Rikvold, Pernille et al. Lactobacillus reuteri and Streptococcus oligofermentans are efficient colonizers in a multi-species model of cariogenic biofilm. 68th ORCA Congress, Zagreb, 07 Jul 2021, Zagreb, Croatia, Conference abstract for conference, 2021.

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@conference{d651df2dca0a4379af897e9c463ebead,
title = "Lactobacillus reuteri and Streptococcus oligofermentans are efficient colonizers in a multi-species model of cariogenic biofilm",
abstract = "Background: The probiotic organisms Lactobacillus reuteri and Streptococcus oligofermentans are known for their inhibitory effect on cariogenic bacteria. Recently, they were shown to reduce Streptococcus mutans counts and lesion depth in a three-species model (S. mutans, Lactobacillus rhamnosus and Actinomyces naeslundii) of dental biofilm [Chen et al.: Microorganisms 2020;8:1272]. Here, we aimed to design and validate specific fluorescence in situ hybridization (FISH) probes for L. reuteri and S. oligofermentans and to investigate how both organisms contribute to biofilm architecture in the model.Materials and methods: Biofilms (n=10) were grown for 10 days on bovine enamel slabs and rinsed daily with either L. reuteri (LR) or S. oligofermentans (SO). Probe sequences (L. reuteri: LAROI1 5{\textquoteright}-GAGAGAGAGAGAG-3{\textquoteright}; S. oligofermentans: SCRI1 5{\textquoteright}- GAGAGAGAGAGAG -3{\textquoteright}) were selected in the hypervariable segment V1 of the 16S rRNA gene and checked for duplex and hairpin formation using Oligo 7. Probe specificity and hybridization conditions were tested in silico using Probe Match and mathFISH. Hybridizations were performed at 30% formamide and 46°C. Fixed cells of L. reuteri, S. oligofermentans, S. mutans, L. rhamnosus and A. naeslundii served as controls. After FISH (n=5), samples were analyzed by confocal microscopy. Total and strain-specific bacterial numbers were assessed by quantitative polymerase chain reaction (n=5). Results: LAROI1 and SCRI1 specifically targeted L. reuteri and S. oligofermentans, respectively, and both organisms constituted an important part of the biofilms. LAROI1, however, competed with EUB338 for binding and could not be used concomitantly. LR biofilms showed significantly reduced numbers of L. rhamnosus (82%) and S. mutans (90%), and S. mutans (94%) and A. naeslundii (58%) were significantly reduced in the SO group (p<0.05). Conclusion: L. reuteri and S. oligofermentans are firmly integrated in multi-species biofilms.",
author = "Pernille Rikvold and Lene Gr{\o}nkj{\ae}r and Zhihui Chen and Gerd G{\"o}stemeyer and Falk Schwendicke and Sebastian Schlafer",
year = "2021",
month = jul,
day = "8",
language = "English",
note = "68th ORCA Congress, Zagreb ; Conference date: 07-07-2021 Through 10-07-2021",

}

RIS

TY - ABST

T1 - Lactobacillus reuteri and Streptococcus oligofermentans are efficient colonizers in a multi-species model of cariogenic biofilm

AU - Rikvold, Pernille

AU - Grønkjær, Lene

AU - Chen, Zhihui

AU - Göstemeyer, Gerd

AU - Schwendicke, Falk

AU - Schlafer, Sebastian

PY - 2021/7/8

Y1 - 2021/7/8

N2 - Background: The probiotic organisms Lactobacillus reuteri and Streptococcus oligofermentans are known for their inhibitory effect on cariogenic bacteria. Recently, they were shown to reduce Streptococcus mutans counts and lesion depth in a three-species model (S. mutans, Lactobacillus rhamnosus and Actinomyces naeslundii) of dental biofilm [Chen et al.: Microorganisms 2020;8:1272]. Here, we aimed to design and validate specific fluorescence in situ hybridization (FISH) probes for L. reuteri and S. oligofermentans and to investigate how both organisms contribute to biofilm architecture in the model.Materials and methods: Biofilms (n=10) were grown for 10 days on bovine enamel slabs and rinsed daily with either L. reuteri (LR) or S. oligofermentans (SO). Probe sequences (L. reuteri: LAROI1 5’-GAGAGAGAGAGAG-3’; S. oligofermentans: SCRI1 5’- GAGAGAGAGAGAG -3’) were selected in the hypervariable segment V1 of the 16S rRNA gene and checked for duplex and hairpin formation using Oligo 7. Probe specificity and hybridization conditions were tested in silico using Probe Match and mathFISH. Hybridizations were performed at 30% formamide and 46°C. Fixed cells of L. reuteri, S. oligofermentans, S. mutans, L. rhamnosus and A. naeslundii served as controls. After FISH (n=5), samples were analyzed by confocal microscopy. Total and strain-specific bacterial numbers were assessed by quantitative polymerase chain reaction (n=5). Results: LAROI1 and SCRI1 specifically targeted L. reuteri and S. oligofermentans, respectively, and both organisms constituted an important part of the biofilms. LAROI1, however, competed with EUB338 for binding and could not be used concomitantly. LR biofilms showed significantly reduced numbers of L. rhamnosus (82%) and S. mutans (90%), and S. mutans (94%) and A. naeslundii (58%) were significantly reduced in the SO group (p<0.05). Conclusion: L. reuteri and S. oligofermentans are firmly integrated in multi-species biofilms.

AB - Background: The probiotic organisms Lactobacillus reuteri and Streptococcus oligofermentans are known for their inhibitory effect on cariogenic bacteria. Recently, they were shown to reduce Streptococcus mutans counts and lesion depth in a three-species model (S. mutans, Lactobacillus rhamnosus and Actinomyces naeslundii) of dental biofilm [Chen et al.: Microorganisms 2020;8:1272]. Here, we aimed to design and validate specific fluorescence in situ hybridization (FISH) probes for L. reuteri and S. oligofermentans and to investigate how both organisms contribute to biofilm architecture in the model.Materials and methods: Biofilms (n=10) were grown for 10 days on bovine enamel slabs and rinsed daily with either L. reuteri (LR) or S. oligofermentans (SO). Probe sequences (L. reuteri: LAROI1 5’-GAGAGAGAGAGAG-3’; S. oligofermentans: SCRI1 5’- GAGAGAGAGAGAG -3’) were selected in the hypervariable segment V1 of the 16S rRNA gene and checked for duplex and hairpin formation using Oligo 7. Probe specificity and hybridization conditions were tested in silico using Probe Match and mathFISH. Hybridizations were performed at 30% formamide and 46°C. Fixed cells of L. reuteri, S. oligofermentans, S. mutans, L. rhamnosus and A. naeslundii served as controls. After FISH (n=5), samples were analyzed by confocal microscopy. Total and strain-specific bacterial numbers were assessed by quantitative polymerase chain reaction (n=5). Results: LAROI1 and SCRI1 specifically targeted L. reuteri and S. oligofermentans, respectively, and both organisms constituted an important part of the biofilms. LAROI1, however, competed with EUB338 for binding and could not be used concomitantly. LR biofilms showed significantly reduced numbers of L. rhamnosus (82%) and S. mutans (90%), and S. mutans (94%) and A. naeslundii (58%) were significantly reduced in the SO group (p<0.05). Conclusion: L. reuteri and S. oligofermentans are firmly integrated in multi-species biofilms.

M3 - Conference abstract for conference

T2 - 68th ORCA Congress, Zagreb

Y2 - 7 July 2021 through 10 July 2021

ER -