Tongue microbiota in relation to the breathing preference in children undergoing orthodontic treatment

doi:10.21203/rs.3.rs-4653787/v1

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Research Article

Tongue microbiota in relation to the breathing preference in children undergoing orthodontic treatment

https://doi.org/10.21203/rs.3.rs-4653787/v1

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Objectives

Mouth breathing (MB) and craniofacial anomalies are interrelated and linked to pediatric obstructive sleep apnea. Both MB and orthodontic therapy can influence the oral microbiome. This study examines the relationship between tongue microbiota and breathing preference in children with orthodontic anomalies.

Materials and Methods

The study included 30 children, 15 with MB and 15 with nose breathing (NB), matched by age, sex, and BMI. They underwent clinical and orthodontic examinations, sleep apnea monitoring, and collection of tongue swabs before (M0) and six months into orthodontic therapy (M6). Oral candidas were analyzed using MALDI-TOF mass spectrometry, and bacteriome analysis was conducted using 16S rRNA sequencing.

Results

At M0, oral candidas were equally present in both groups. At M6, Candida sp. were found in six children with MB but in none with NB (p = 0.01). No significant differences in bacterial diversity were observed between groups. However, children with MB had higher relative abundance of Solobacterium than others (q = 0.01).

Conclusions

Oral candidas are more common during orthodontic therapy in children with MB. While the study didn't find broad impacts of MB on oral bacteriome composition, it identified a significant link between Solobacterium and MB, highlighting the importance of managing halitosis in these patients.

tongue microbiota

mouth breathing

orthodontic treatment

children

pediatric sleep apnea

craniofacial anomaly

halitosis

Solobacterium

oral candida

The importance of physiological breathing, i.e., nose breathing preference (NB), is a frequently discussed topic. Mouth breathing preference (MB) occurs in the case of nasal obstruction or habit and may subsequently influence the growth and development of the craniofacial region in growing children, which may be expressed in different forms and characteristics. ^1,2

In children with MB, an insufficient lip seal and open mouth, narrow, constricted hard palate, hyperdivergent growth pattern of the mandible, and disproportion in the sagittal plane, as well as elongation of the lower third of the face are often found.^2,3 Some of these characteristics are known to be risk factors for the narrowing of the upper airways and their possible collapse, especially during sleep, and are often found in children suffering from pediatric obstructive sleep apnea (POSA). ^4,5 Another explanation of the co-occurrence of MB and POSA may lie in the fact that some mechanisms known to facilitate MB (such as tongue protrusion or mandibular advancement) cannot be applied in sleep, as they are active processes. This may also contribute to an increased risk of a collapse of the upper airway and apnea.⁶

A strong preference for MB, which according to literature may be found in 50–56% of the pediatric population, leads to changes in the quality and quantity of saliva, water imbalance and mucosal moisture, influencing the composition of the oral microbiome.⁷ Not only these changes and dryness of the surfaces but also dental caries, periodontal diseases, oronasal infections, and allergies, all directly or indirectly associated with MB, contribute to halitosis.⁸ Halitosis, known also as bad breath or oral malodor, manifest as an unpleasant odor from the mouth, which is mainly caused by putrefactive actions of microorganisms.⁹ Several studies have reported increased halitosis in children with the MB. ^10,11 On the other hand, the number of studies published so far focused on the oral microbiota in children with MB and/or POSA is limited. ^7,12–14

This complex interplay between the oral microbiome, MB, orthodontic anomalies, orthodontic therapy, and POSA underscores the importance of a this study. We hypothesized that tongue microbiota in children with MB differ from those with NB. The presented study aimed to analyze the bacteriomes of the tongue's dorsa in children with MB/NB and orthodontic anomalies undergoing orthodontic treatment who are, therefore, at high POSA risk. We have previously observed changes in the quantity of specific oral bacteria and candidas in the gingival crevicular fluid (GCF) and dental plaque of children undergoing orthodontic therapy ¹⁵; in view of these results, we planned to investigate tongue microbiota also in relation to the absence/presence of orthodontic appliances.

Design and clinical examination

In this pilot prospective case-control study, a meticulous methodology was employed to investigate the oral microbiota in a specific group of 30 children aged 7–12 years from the Czech Republic. From the participants’ pool, 15 children with MB (cases) were selected and matched with 15 children with NB (controls) according to their age, sex, and BMI.

All children were selected from the pool of patients from the Orthodontic Department of the Clinic of Stomatology, St. Anne’s University Hospital, Brno, Czech Republic. Our inclusion criteria were: age 7 to 12 years, Czech or Slovak nationality, and parents/caregivers willing to sign an informed consent for the participation of their child in the study. These criteria were approached to participate in our study prior to the start of their orthodontic treatment. The exclusion criteria were: a diagnosis of a genetically determined syndromic disease associated with craniofacial dysmorphism, such as Down syndrome or the Pierre-Robin sequence, and the use of antibiotics within 2 months prior to the procedure.

Children meeting these criteria underwent a complex orthodontic examination, including the extraoral and intraoral examination, photographs, dental impressions, and X-rays (cephalogram and orthopantomogram). During this initial examination, we recorded not only the presence of orthodontic anomalies but also the overall habitus of the patient and their breathing pattern. The preference for mouth or nose breathing was based on the complex patient history taken from the parent of the patient, visual assessment of the lip posture, and clinical examination of the nasal movements. All children were also referred for an ear, nose and throat (ENT) examination for the endoscopic examination of the nose and nasopharynx; however, only 16 patients were willing to undergo this examination. The patients’ breathing pattern was also examined during this ENT examination; the emphasis was, however, put on their medical history as we were interested also in nocturnal MB.

All patients underwent home sleep apnea examination using the Alice OneNight device, Philips Respironics (Murrysville, USA). The quality of their sleep-breathing and the risk of POSA was established based on the analysis from the home sleep apnea monitoring using the scoring officially recommended for children under 12 years, i.e.: AHI < 1 (physiological finding), AHI ≥ 1 to < 5 (mild POSA), AHI ≥ 5 to < 10 (moderate POSA), or AHI ≥ 10 (severe POSA) ).¹⁶

The preparation for sample collection required stringent standardization, with participants and their parents receiving clear instructions not to eat, chew chewing gums, drink anything besides water, or perform any oral hygiene for 1 hour before the procedure. The collection procedure was scheduled at two distinct intervals: before the beginning of the orthodontic treatment (timepoint M0) and 6months after the treatment with the orthodontic appliance started (timepoint M6).

The information about the sampling and analysis of oral candidas and bacteriome, including the process of DNA extraction and 16S rRNA amplicon sequencing, as well as all the bioinformatic and statistical analysis, is closely described in the Supplement.

Children in our study were matched according to their age, sex, and BMI (z-score). Therefore, there were no statistically significant differences between cases and controls (p > 0.05 for all). Distributions of the presence of maxillary constriction, ANB angle, tongue range of motion ratio (TRMR), crowding, overjet, grade of adenoid vegetation, and risk of POSA (according to AHI) were also similar between studied groups (p > 0.05 for all). However, the median AHI was found higher in the MB group than in the NB group (2.2 vs. 1.1, respectively, p = 0.036). Eight children from the MB group were treated with removable orthodontic appliances and seven with a fixed one, whereas in the NB group, the majority was treated with fixed appliances; in relation to the type of the orthodontic appliance (fixed vs. removable appliance), no significant difference between studied groups in AHI was observed (p > 0.05), see Table 1.

Table 1

Demographic data of studied children (N = 30)
	Mouth breathing (N = 15)	Nose breathing (N = 15)	p-value
Age
Median [Min; Max]	9.0 [7.0; 12)	10 [7.0; 12)	0.395
Mean (± SD)	9.5 (± 1.6)	10 (± 1.8)
Sex
male	6 (40%)	5 (33%)	1
BMI (z-score)
Median [Min; Max]	0.73 [-2.8; 1.9)	-0.30 [-2.7; 3.0)	0.412
Mean (± SD)	-0.0031 (± 1.4)	-0.20 (± 1.3)
Maxillary constriction	6 (40%)	9 (60%)	0.465
Median [Min; Max]	4.1 [1.3; 7.2)	3.5 [0.10; 5.9)	0.206
Mean (± SD)	4.2 (± 1.8)	3.3 (± 1.7)
ANB angle
Median [Min; Max]	4.5 [-1.4; 9.8)	2.7 [-1.9; 5.9)	0.329
Mean (± SD)	4.3 (± 3.2)	3.0 (± 2.1)
TRMR
Median [Min; Max]	0.67 [0.55; 0.96)	0.77 [0.48; 0.95)	0.184
Mean (± SD)	0.68 (± 0.12)	0.74 (± 0.14)
Crowding	8 (53%)	6 (40%)	0.714
Overjet	11 (73%)	10 (67%)	1
Removable orthodontic appliance	8 (53%)	4 (27%)	0.264
Fixed orthodontic appliances	7 (47%)	11 (73%)
AHI
Median [Min; Max]	2.2 [0.50; 7.5)	1.1 [0; 3.2)	0.036
Mean (± SD)	2.6 (± 1.7)	1.4 (± 1.1)
AHI < 1 (physiological finding)	2 (13%)	6 (40%)	0.180
AHI ≥ 1 to < 5 (mild POSA)	12 (80%)	9 (60%)
AHI ≥ 5 to < 10 (moderate POSA)	1 (7%)	0 (0%)
Adenoid vegetation
Grade I	6 (40%)	6 (40%)	1
Grade II	2 (13%)	2 (13%)
Missing	7 (47%)	7 (47%)
Candida sp. in timepoint M0	1 (7%)	1 (7%)	1
Candida sp. in timepoint M6	6 (40%)**	0 (0%)	0.012
N, number of patients; SD, standard deviation; BMI, body mass index; ANB angle, angle between the points A, nasion and B; TRMR, tongue range motion ratio; AHI, apnea-hypopnea index; POSA, pediatric obstructive sleep apnea; M0, before orthodontic treatment; M6, till the end of 6th month of orthodontic treatment; ** data from the analysis are missing in 2 patients

Candida sp. was found only in one patient in each group before the orthodontic treatment (M0, p > 0.05). However, during the orthodontic treatment (M6), Candida sp. were identified in six children with MB and in none of the children with NB (p = 0.012), see Table 1. Dominantly, Candida albicans was identified, in one case together with Candida dubliniensis, and another case Candida lusitaniae.

After quality filtering and chimeras removal, 799,219 reads (median = 10,411 reads per sample; interquartile range, IQR = 10,066) originated from patients’ samples and 184,081 reads (median = 13,827, IQR = 6,968) from NC. After filtering out ASV unassigned at the phylum level as well as cyanobacteria and mitochondria, most phyla (8/12) were present at low abundances (sum of reads < 5% of total). Firmicutes, Proteobacteria, Bacteroidota, and Actinobacteriota, were the most abundant phyla, comprising in median 63.1%, 12.5%, and 9.2% of phyla per patient’s sample, respectively.

No significant differences in alpha diversities of bacterial genera were detected between the MB and NB groups before or during the treatment (p > 0.05 for all). However, patient samples from each group were significantly more diverse in the number of amplicon sequence variants (ASVs) than NC and their alpha diversities (Shannon index) were also higher than in NC (p < 0.001 for all), see Figs. 1 and 2.

Similarly, as in the case of alpha diversity, there were no differences in beta diversities among patient´s groups and timepoints (p > 0.05 for all, with the exception of NC), see Fig. 3. Streptococcus, Prevotella 7, Veilonella, Neisseria, Rothia, and Haemophillus were the most abundant genera, see the heatmap in Fig. 4. However, Streptococcus, Prevotella 7, Veilonella, Neisseria, and Rothia were present also in NC. Eleven bacterial genera were found exclusively in NC (Flavobacterium, Acinetobacter, Ligilactobacillus, Lactobacillus, Chlamydiaceae, Bifidobacterium, Paracoccus, Proteiniclasticum, Faecalibacterium, Oligoflexaceae, Butyricicoccus). On the other hand, five genera (Haemophillus, Actinomyces, Alloprevotella, Fusobacterium, and Prevotella) were found only in patients’ samples and not in NC.

At the level of genera, there were almost no differences in relative abundances of individual genera among patient groups and timepoints (q > 0.05). However, before the start of the orthodontic therapy, we observed a significantly higher relative abundance of the Solobacterium genus in children with MB than in children with NB (q = 0.012, Dunn's test), see Fig. 5.

In the present study, the differences in bacterial diversity and composition between children with orthodontic anomalies undergoing orthodontic treatment were investigated according to their breathing preferences. Our results are in contrast to the expectations set by previous research highlighting a significant difference in beta diversity and microbial profiles between patients with POSA and/or MB and controls.^17,18 Even though this information is not in agreement with our observations, it's worth considering that in our group of patients, even though some of them were diagnosed with higher POSA risk based on home sleep apnea test and AHI score, they were all

with AHI below 10.

Studies on microbial diversity in children with POSA have noted a significant difference in beta diversity; Haemophilus, Fusobacterium, and Porphyromonas were found in higher abundances in samples collected from adenoids and tonsils of patients with POSA than in controls.^7,12 POSA also affects the microbiota composition of the buccal mucosa, altering levels of Firmicutes, Proteobacteria, Bacteroidetes, Fusobacteria, and Actinobacteria.¹⁷

MB is often found in children with POSA but also in otherwise healthy subjects, further influencing the oral and nasopharyngeal microbiota, enriching bacteria with pathogenic potential, such as Acinetobacter sp. in supragingival plaque, Neisseria sp. in unstimulated saliva, Streptococcus pneumoniae in the pharynx, and Stenotrophomonas sp. in the nostrils.⁷ MB has been also linked to chronic gingival inflammation and higher plaque index.¹⁸ The tonsillar microbiota in patients with POSA differs, with a greater presence of genera such as Porphyromonas, Moraxella and Corynebacteria in patients with POSA and adenotonsillitis and higher prevalence of Leptotrichia, Campolybacter or Paludibacter in children with POSA and adenotonsillar hypertrophy.¹³ The surfaces of the adenoids and palatine tonsils harbor bacterial growth that may contribute to mucosal infection and influence POSA or recurrent tonsillitis.¹⁴ Differences in abundance include higher abundances of Parvimonas, Prevotella, and Treponema in patients with recurrent tonsillitis, and Haemophilus and Capnocytophaga in patients with POSA, with a predominance of Proteobacteria and Firmicutes in sleep-disordered breathing.¹⁴

The oral microbiome houses hundreds of microbial species with a dominance of Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria, Spirochaetes, and Fusobacteria.¹⁹ Disruption of homeostasis in the oral microbiota can lead to a wide range of problems, including caries, gum inflammations, aphthae, halitosis, etc.²⁰

Halitosis, or bad breath, is caused by volatile compounds produced mainly by anaerobic bacteria, such as sulfur compounds, ketones, sulfides, aldehydes, and amines. The tongue with its papillae and anaerobic sites offers a suitable environment to anaerobic bacteria causing halitosis.²⁰ In healthy subjects without halitosis, Streptococcus salivarius is the predominant species; in patients with halitosis, however, it was typically absent and the microbiota differed from healthy patients, with Solobacterium moorei, Atopobium parvulum, Eubacterium sulci, and Fusobacterium periodonticum showing the strongest associations with halitosis.²¹

The higher relative abundance of Solobacterium in children with MB compared to children with NB is a significant result of our research. Solobactertium is a bacterial genus that has been previously associated with halitosis. ^22,23 It thrives in anaerobic conditions, such as those found on the dorsum of the tongue or in the periodontal pockets, and is known for its sulfurous metabolic byproducts that contribute to bad breath.^21–24

Our study uniquely identifies the specific relationship between Solobacterium and MB. MB can often lead to a dry oral environment, thus reducing the self-cleaning ability of the oral cavity, increasing plaque accumulation, and facilitating bacterial colonization. Therefore, dry mouth, known also as xerostomia, disrupts the natural balance of the oral environment.²⁵ This imbalance may result in an overgrowth of anaerobic bacteria, such as Solobacterium moorei, which may further intensify the problem of bad breath as it creates a favorable environment for these bacteria to decompose cellular debris and proteins, leading to the production of malodorous compounds. ^22–24 Thus, MB may not only contribute to the conditions that favor the growth of Solobacterium, but can also intensify the symptoms of halitosis that the bacterium causes. This finding not only strengthens the link between specific bacterial genera and MB but also implies a potential pathophysiological role for Solobacterium in mouth breathers.

Multiple studies reported changes in the oral microflora during orthodontic therapy; however, most of them investigated saliva or dental plaque samples. ^26,27 In our recently published study, we emphasized the importance of the complex perspective in the research of the dynamics of the oral ecosystem. .¹⁵ We found that in patients whose plaque index deteriorated in the medium term after the bonding of the appliances (before the end of the 7th month of the treatment), the probability of finding any of the seven selected periodontal bacterial strains combined with oral candidas in GCF/dental plaque samples was 10 times higher than before their orthodontic therapy.¹⁵

The main limitation of our study is the process of diagnosis of the MB. There are several approaches to diagnosing the mouth-breathing pattern, including mirror and/or water retention tests. ²⁸ In our study, we have used visual assessment of the lip seal, clinical examination of the nasal movements and, most importantly, complex medical history was taken from the patient’s parent. The latter is often used for diagnosing mouth breathing; ²⁹ however, Costa et al. stated that such investigation may be insufficient and that children should be referred for an ENT examination³⁰, which we weren’t able to achieve in all our patients, even though they have been all referred to an ENT specialist. Still, we can consider our pilot study methodologically strong because we have matched groups with many clinical parameters observed and examined across various medical fields. Moreover, the conditions for sample collection were very strict and the samples were analyzed using state-of-the-art approaches.

In conclusion, while our findings did not confirm some of the previously reported associations between mouth breathing preference, orthodontic anomalies, and changes in oral microbiota diversity and composition, they introduced a noteworthy connection between the oral presence of Solobacterium as well as oral candidas and mouth breathing preference. This discovery adds a valuable dimension to our understanding of the complex relationship between the oral microbiome and respiratory mode and presents opportunities for further exploration, especially in the context of halitosis management. It also emphasizes the need for continued investigation, potentially incorporating longitudinal studies and a more diverse patient population, to deepen our understanding and develop effective diagnostic and therapeutic strategies based on these nuanced insights.

Ethics approval and consent to participate

The study was performed with the approval of the Ethics Committee of the St. Anne’s Faculty Hospital (No. 33V/2020; date: 10^th June, 2020). Written informed consent was obtained from all parents or caregivers of participating children in line with the Declaration of Helsinki before inclusion in the study.

Consent for publication

Not applicable.

Availability of data and materials

All data generated or analysed during this study are included in this published article [and its supplementary information files].

Conflict of interest

The authors declare no competing interests.

Author contribution

ZMV and PBL designed the study, data analysis, interpretation of results, and drafted the paper; ZMV was responsible for the clinical part of the study; PB, LB, and PBL designed and/or carried out the molecular analysis; JB provided data analysis; JB and PBL designed and/or created figures; FR was responsible for the oral candida analysis and critically reviewed the manuscript. All authors revised the final version of the manuscript. Authors also confirm that they have no conflict of interests

Acknowledgments

The study was supported by the Ministry of Health of the Czech Republic, grants No. NU20-08-00205, and by a project funded by the University Hospital Brno, Ministry of Health, Czech Republic – RVO (FNBr, 65269705). This publication was supported from the European Union’s Horizon 2020 Research and Innovation Programme under grant agreement No. 857560. This publication reflects only the author´s view and the European Commission is not responsible for any use that may be made of the information it contains. The authors also thank the Research Infrastructure RECETOX RI (No. LM2023069) and the project CETOCOEN EXCELLENCE (No CZ.02.1.01/0.0/0.0/17_043/0009632) financed by the Ministry of Education, Youth and Sports for supportive background.

We thank Pavel Hornik for the ENT examinations of children enrolled in the study. We also thank Vojtech Barton for the bioinformatic analysis and Jaroslav Janosek for his valuable comments.

Basheer B, Hegde KS, Bhat SS, Umar D, Baroudi K. Influence of mouth breathing on the dentofacial growth of children: a cephalometric study. J Int Oral Health. 2014;6(6):50–5.
Lessa FC, Enoki C, Feres MF, Valera FC, Lima WT, Matsumoto MA. Breathing mode influence in craniofacial development. Braz J Otorhinolaryngol. 2005;71(2):156–60. 10.1016/s1808-8694(15)31304-5.
Drevensek M, Papić JS. The influence of the respiration disturbances on the growth and development of the orofacial complex. Coll Antropol. 2005;29(1):221–5.
Johal A, Patel SI, Battagel JM. The relationship between craniofacial anatomy and obstructive sleep apnoea: a case-controlled study. J Sleep Res. 2007;16(3):319–26. 10.1111/j.1365-2869.2007.00599.x.
Marino A, Malagnino I, Ranieri R, Villa MP, Malagola C. Craniofacial morphology in preschool children with obstructive sleep apnoea syndrome. Eur J Paediatr Dent. 2009;10(4):181–4.
Harvold EP, Tomer BS, Vargervik K, Chierici G. Primate experiments on oral respiration. Am J Orthod. 1981;79(4):359–72. 10.1016/0002-9416(81)90379-1.
Fan C, Guo L, Gu H, Huo Y, Lin H. Alterations in Oral-Nasal-Pharyngeal Microbiota and Salivary Proteins in Mouth-Breathing Children. Front Microbiol. 2020;11:575550. Published 2020 Oct 9. 10.3389/fmicb.2020.575550.
Triana BEG, Ali AH, León IG. Mouth breathing and its relationship to some oral and medical conditions: physiopathological mechanisms involved. Revista Habanera de Ciencias Médicas. 2016;15(2):200–12.
Pieniążek A, Pietrzak M. Halitoza-etiologia, metody diagnostyki i leczenie. Halitosis-etiology, methods of diagnosis and treatment. J Health Study Med. 2017;2:101–22.
Bawazir OA. Risk Factors, Diagnosis, and Management of Halitosis in Children: A Comprehensive Review. J Contemp Dent Pract. 2021;22(8):959–63. Published 2021 Aug 1.
Motta LJ, Bachiega JC, Guedes CC, Laranja LT, Bussadori SK. Association between halitosis and mouth breathing in children. Clin (Sao Paulo). 2011;66(6):939–42. 10.1590/s1807-59322011000600003.
Davidovich E, Hevroni A, Gadassi LT, Spierer-Weil A, Yitschaky O, Polak D. Dental, oral pH, orthodontic and salivary values in children with obstructive sleep apnea. Clin Oral Investig. 2022;26(3):2503–11. 10.1007/s00784-021-04218-7.
Galli J, Calò L, Posteraro B, et al. Pediatric oropharyngeal microbiome: Mapping in chronic tonsillitis and tonsillar hypertrophy. Int J Pediatr Otorhinolaryngol. 2020;139:110478. 10.1016/j.ijporl.2020.110478v.
Johnston J, Hoggard M, Biswas K, et al. The bacterial community and local lymphocyte response are markedly different in patients with recurrent tonsillitis compared to obstructive sleep apnoea. Int J Pediatr Otorhinolaryngol. 2018;113:281–8. 10.1016/j.ijporl.2018.07.041.
Marincak Vrankova Z, Rousi M, Cvanova M, et al. Effect of fixed orthodontic appliances on gingival status and oral microbiota: a pilot study. BMC Oral Health. 2022;22(1):455. 10.1186/s12903-022-02511-9. Published 2022 Oct 27.
Savini S, Ciorba A, Bianchini C, et al. Assessment of obstructive sleep apnoea (OSA) in children: an update. Acta Otorhinolaryngol Ital. 2019;39(5):289–97. 10.14639/0392-100X-N0262.
Xu H, Li X, Zheng X, et al. Pediatric Obstructive Sleep Apnea is Associated With Changes in the Oral Microbiome and Urinary Metabolomics Profile: A Pilot Study. J Clin Sleep Med. 2018;14(9):1559–67. 10.5664/jcsm.7336. Published 2018 Sep 15.
Sharma RK, Bhatia A, Tewari S, Narula SC. Distribution of gingival inflammation in mouth breathing patients: an observational pilot study. J Dentistry Indonesia. 2016;23(2):28–32.
Dewhirst FE, Chen T, Izard J, et al. The human oral microbiome. J Bacteriol Oct. 2010;192(19):5002–17. 10.1128/jb.00542-10.
Deo PN, Deshmukh R. Oral microbiome: Unveiling the fundamentals. J Oral Maxillofac Pathol. 2019;23(1):122–8. 10.4103/jomfp.JOMFP_304_18v.
Kazor CE, Mitchell PM, Lee AM, et al. Diversity of bacterial populations on the tongue dorsa of patients with halitosis and healthy patients. J Clin Microbiol. 2003;41(2):558–63. 10.1128/JCM.41.2.558-563.2003.
Haraszthy VI, Gerber D, Clark B, et al. Characterization and prevalence of Solobacterium moorei associated with oral halitosis. J Breath Res. 2008;2(1):017002. 10.1088/1752-7155/2/1/017002.
Barrak I, Stájer A, Gajdács M, Urbán E. Small, but smelly: the importance of Solobacterium moorei in halitosis and other human infections. Heliyon. 2020;6(10):e05371. 10.1016/j.heliyon.2020.e05371. Published 2020 Oct 29.
Haraszthy VI, Zambon JJ, Sreenivasan PK, et al. Identification of oral bacterial species associated with halitosis. J Am Dent Assoc. 2007;138(8):1113–20. 10.14219/jada.archive.2007.0325.
Ligtenberg AJ, Almståhl A. Xerostomia and the oral microflora. Dry mouth: A clinical guide on causes, effects and treatments. 2015:81–101.
Bergamo AZN, de Oliveira KMH, Matsumoto MAN, et al. Orthodontic appliances did not increase risk of dental caries and periodontal disease under preventive protocol. Angle Orthod. 2019;89(1):25–32. 10.2319/022118-139.1.
Rodríguez-Rentería M, Márquez-Preciado R, Ortiz-Magdaleno M, Bermeo-Escalona J, Sánchez-Vargas LO. Frequency of Pathogenic Microorganisms in Removable Orthodontic Appliances and Oral Mucosa in Children. J Clin Pediatr Dent. 2021;45(2):135–9. 10.17796/1053-4625-45.2.11.
de Menezes VA, Barbosa Leal R, Motta Moura M, Granville-Garcia AF. Influence of socio-economic and demographic factors in determining breathing patterns: a pilot study. Braz J Otorhinolaryngol. 2007;73(6):826–34. 10.1016/S1808-8694(15)31179-4.
Pacheco MC, Casagrande CF, Teixeira LP, Finck NS, de Araújo MT. Guidelines proposal for clinical recognition of mouth breathing children. Dent Press J Orthod. 2015;20(4):39–44. 10.1590/2176-9451.20.4.039-044.oar.
Costa JG, Costa GS, Costa C, Vilella OV, Mattos CT, Cury-Saramago AA. Clinical recognition of mouth breathers by orthodontists: A preliminary study. Am J Orthod Dentofac Orthop. 2017;152(5):646–53. 10.1016/j.ajodo.2017.03.025.

No competing interests reported.

SupplementBMC.docx
Supplement: Sampling and analysis of oral candidas; Sampling for bacteriome analysis; DNA extraction and 16S rRNA amplicon sequencing; Bioinformatic analysis of data from 16S rRNA amplicon sequencing; Statistical analysis

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Tongue microbiota in relation to the breathing preference in children undergoing orthodontic treatment

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