Relationship between Phenotypic and Genotypic Resistance of Subgingival Biofilm Samples in Patients with Periodontitis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Patients
2.2. Sampling Process
2.3. Microbiological Growth and Identification of Isolates
2.4. ARG Testing
2.5. Bioinformatics
2.6. Predictive Values
2.7. Comparison of Shotgun Analysis and MALDI-TOF-MS/16S Sequence Analysis
3. Results
3.1. Isolated Bacterial Strains
3.2. Prevalence of ARGs
3.3. Predictive Values of Resistance Genes
3.4. Identification via Shotgun Analysis
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Antibiotics | Concentration in Agar Plates (mg/L) |
---|---|
Ampicillin-Sulbactam | 2; 4 and 8 |
Clindamycin | 2 and 4 |
Doxycycline | 2 and 4 |
Metronidazole | 2 and 4 |
Parameter | Data (Mean ± SD, n = 19) |
---|---|
Age (years) | 56.26 ± 13.45 |
Sex (male/female, %) | 47/53 |
Smoker (%) | 32 |
PD (sites) * | 6.16 ± 1.73 |
CAL (sites) * | 5.63 ± 2.89 |
BOP (positive/negative, sites, %) * | 58/42 |
Taxonomic Rank | n | % | |
---|---|---|---|
Genus | Species | ||
Streptococcus | S. mitis/oralis | 5 | 26.32 |
S. constellatus | 4 | 21.05 | |
S. anginosus | 3 | 15.79 | |
S. intermedius | 2 | 10.53 | |
S. massiliensis | 1 | 5.26 | |
S. gordonii | 1 | 5.26 | |
Prevotella | P. maculosa | 4 | 21.05 |
P. nigrescens | 2 | 10.53 | |
P. veroralis | 1 | 5.26 | |
P. salivae | 1 | 5.26 | |
P. nanceiensis | 1 | 5.26 | |
P. intermedia | 1 | 5.26 | |
P. buccae | 1 | 5.26 | |
Capnocytophaga | C. gingivalis | 6 | 31.58 |
C. ochracea | 2 | 10.53 | |
C. granulosa | 1 | 5.26 | |
Gemella | G. morbillorum | 6 | 31.58 |
Actinomyces | A. meyeri | 3 | 15.79 |
A. odontolyticus | 2 | 10.53 | |
Bacteroides | B. thetaiotaomicron | 1 | 5.26 |
B. pyogenes | 1 | 5.26 | |
B. ovatus/xylanisolvens | 1 | 5.26 | |
Anaplasma | A. ovis | 1 | 5.26 |
A. phagocytophilum | 1 | 5.26 | |
Cutibacterium | C. acnes | 2 | 10.53 |
Solobacterium | S. moorei | 2 | 10.53 |
Alloscardovia | A. omnicolens | 1 | 5.26 |
Corynebacterium | C. durum | 1 | 5.26 |
Microbacterium | M. flavescens/laevaniformans | 1 | 5.26 |
Bulleida | B. extructa | 1 | 5.26 |
Eggerthia | E. catenaformis | 1 | 5.26 |
Bacillus | B. cereus group | 1 | 5.26 |
Fusobacterium | F. nucleatum | 1 | 5.26 |
Antibiotic | Resistance Genes | Number of Biofilm Samples |
---|---|---|
Ampicillin-Sulbactam | cfxA | 3 |
cfxA3 | 4 | |
cfxA4 | 3 | |
cfxA5 | 3 | |
Clindamycin | ermB | 2 |
ermF | 5 | |
ermG | 1 | |
mefA | 3 | |
Doxycycline | tet32 | 4 |
tetM | 3 | |
tetO | 1 | |
tetQ | 2 | |
Metronidazole | nimI | 0 |
Sample | Resistance Genes | Phenotypic Resistance of the Samples | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Ampicillin-Sulbactam | Clindamycin | Doxycycline | Metronidazole | Ampicillin-Sulbactam | Clindamycin | Doxycycline | Metronidazole | ||||||
2 mg/L (S) | 4 mg/L (S) | 8 mg/L (R) | 2 mg/L (S) | 4 mg/L (R) | 2 mg/L | 4 mg/L | 2 mg/L (S) | 4 mg/L (R) | |||||
1 | 0 | ermF | 0 | 0 | ø | ø | ø | + | + | ø | ø | + | ø |
2 | cfxA, 3, 4, 5 | ermF | 0 | 0 | ø | ø | ø | ø | + | ø | ø | + | + |
3 | 0 | 0 | 0 | 0 | ø | ø | ø | + | + | + | + | + | + |
4 | 0 | 0 | 0 | 0 | ø | ø | ø | ø | + | ø | ø | + | + |
5 | cfxA3 | erm(B, F, G), mefA | tet32, tetM | 0 | + | + | + | + | + | + | + | + | + |
6 | 0 | 0 | 0 | 0 | ø | ø | ø | ø | ø | ø | ø | + | + |
7 | 0 | mefA | 0 | 0 | ø | ø | ø | + | + | + | ø | + | + |
8 | 0 | ermF | 0 | 0 | ø | ø | ø | + | + | ø | ø | + | + |
9 | 0 | ermF | tet32 | 0 | ø | ø | ø | + | + | ø | ø | + | + |
10 | 0 | 0 | 0 | 0 | ø | ø | ø | + | + | ø | ø | + | + |
11 | 0 | mefA | tet32 | 0 | ø | ø | ø | + | + | + | ø | + | + |
12 | cfxA, 3, 4, 5 | 0 | tet32, tetQ | 0 | ø | ø | ø | ø | ø | ø | ø | + | + |
13 | 0 | ermB | tetM, tetO | 0 | ø | ø | ø | + | + | + | + | + | + |
14 | 0 | 0 | tetM | 0 | ø | ø | ø | + | + | + | ø | + | + |
15 | 0 | 0 | tetQ | 0 | ø | ø | ø | ø | ø | ø | ø | + | + |
16 | cfxA, 3, 4, 5 | 0 | 0 | 0 | ø | ø | ø | ø | ø | ø | ø | + | + |
17 | 0 | 0 | 0 | 0 | ø | ø | ø | + | ø | ø | ø | + | ø |
18 | 0 | 0 | 0 | 0 | ø | ø | ø | + | + | + | ø | + | + |
19 | 0 | 0 | 0 | 0 | ø | ø | ø | ø | ø | ø | ø | + | + |
Sample | Resistance Genes | Identification via MALDI-TOF-MS/16S Sequencing | Identification via Shotgun Analysis | Results |
---|---|---|---|---|
1 | ermF | Prevotella intermedia | Prevotella intermedia | same species |
2 | ermF | Prevotella buccae | Prevotella nigrescens | same genus |
5 | ermF | Bacteroides thetaiotaomicron, Bacteroides ovatus | Bacteroides ovatus | same species |
7 | mefA | Gemella morbillorum, Anaplasma phagocytophilum | Gemella sanguinis | same genus |
8 | ermF | Prevotella maculosa, Corynebacterium durum | Prevotella melaninogenica | same genus |
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Sparbrod, M.; Gager, Y.; Koehler, A.-K.; Jentsch, H.; Stingu, C.-S. Relationship between Phenotypic and Genotypic Resistance of Subgingival Biofilm Samples in Patients with Periodontitis. Antibiotics 2023, 12, 68. https://doi.org/10.3390/antibiotics12010068
Sparbrod M, Gager Y, Koehler A-K, Jentsch H, Stingu C-S. Relationship between Phenotypic and Genotypic Resistance of Subgingival Biofilm Samples in Patients with Periodontitis. Antibiotics. 2023; 12(1):68. https://doi.org/10.3390/antibiotics12010068
Chicago/Turabian StyleSparbrod, Moritz, Yann Gager, Anne-Katrin Koehler, Holger Jentsch, and Catalina-Suzana Stingu. 2023. "Relationship between Phenotypic and Genotypic Resistance of Subgingival Biofilm Samples in Patients with Periodontitis" Antibiotics 12, no. 1: 68. https://doi.org/10.3390/antibiotics12010068