The effect of daily usage of Listerine Cool Mint mouthwash on the oropharyngeal microbiome: a substudy of the PReGo trial.

J G E Laumen, C Van Dijck, S S Manoharan-Basil, T de Block, S Abdellati, B B Xavier, S Malhotra-Kumar, C Kenyon
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Abstract

Introduction. ListerineÒ is a bactericidal mouthwash widely used to prevent oral health problems such as dental plaque and gingivitis. However, whether it promotes or undermines a healthy oral microbiome is unclear.Hypothesis/Gap Statement. We hypothesized that the daily use of Listerine Cool Mint would have a significant impact on the oropharyngeal microbiome.Aim. We aimed to assess if daily usage of Listerine Cool Mint influenced the composition of the pharyngeal microbiome.Methodology. The current microbiome substudy is part of the Preventing Resistance in Gonorrhoea trial. This was a double-blind single-centre, crossover, randomized controlled trial of antibacterial versus placebo mouthwash to reduce the incidence of gonorrhoea/chlamydia/syphilis in men who have sex with men (MSM) taking HIV pre-exposure prophylaxis (PrEP). Fifty-nine MSM taking HIV PrEP were enrolled. In this crossover trial, participants received 3 months of daily Listerine followed by 3 months of placebo mouthwash or vice versa. Oropharyngeal swabs were taken at baseline and after 3 months use of each mouthwash. DNA was extracted for shotgun metagenomic sequencing (Illumina Inc.). Non-host reads were taxonomically classified with MiniKraken and Bracken. The alpha and beta diversity indices were compared between baseline and after each mouthwash use. Differentially abundant bacterial taxa were identified using ANOVA-like differential expression analysis.Results. Streptococcus was the most abundant genus in most samples (n = 103, 61.7 %) with a median relative abundance of 31.5% (IQR 20.6-44.8), followed by Prevotella [13.5% (IQR 4.8-22.6)] and Veillonella [10.0% (IQR 4.0-16.8)]. Compared to baseline, the composition of the oral microbiome at the genus level (beta diversity) was significantly different after 3 months of Listerine (P = 0.006, pseudo-F = 2.29) or placebo (P = 0.003, pseudo-F = 2.49, permutational multivariate analysis of variance) use. Fusobacterium nucleatum and Streptococcus anginosus were significantly more abundant after Listerine use compared to baseline.Conclusion. Listerine use was associated with an increased abundance of common oral opportunistic bacteria previously reported to be enriched in periodontal diseases, oesophageal and colorectal cancer, and systemic diseases. These findings suggest that the regular use of Listerine mouthwash should be carefully considered.

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每天使用李施德林薄荷漱口水对口咽微生物组的影响:PReGo 试验的子研究。
简介李施德林Ò是一种杀菌漱口水,被广泛用于预防牙菌斑和牙龈炎等口腔健康问题。然而,它是促进还是破坏健康的口腔微生物群还不清楚。我们假设每天使用李施德林清凉薄荷漱口水会对口咽微生物群产生重大影响。我们旨在评估每天使用李施德林清凉薄荷是否会影响咽部微生物组的组成。目前的微生物组子研究是预防淋病耐药性试验的一部分。这是一项双盲、单中心、交叉、随机对照试验,目的是通过抗菌漱口水与安慰剂的对比,降低服用艾滋病暴露前预防药物(PrEP)的男男性行为者(MSM)的淋病/衣原体/梅毒发病率。59 名男男性行为者正在服用 HIV PrEP。在这项交叉试验中,参与者每天使用李施德林漱口水 3 个月,然后再使用安慰剂漱口水 3 个月,反之亦然。在基线和使用每种漱口水 3 个月后,分别采集口咽拭子。提取 DNA 进行射枪元基因组测序(Illumina 公司)。用 MiniKraken 和 Bracken 对非宿主读数进行分类。比较了基线和每次使用漱口水后的α和β多样性指数。使用类似方差分析的差异表达分析确定了差异丰富的细菌类群。链球菌是大多数样本中含量最高的菌属(n = 103,61.7%),相对含量中位数为 31.5%(IQR 20.6-44.8),其次是普雷沃特氏菌[13.5%(IQR 4.8-22.6)]和维氏菌[10.0%(IQR 4.0-16.8)]。与基线相比,使用李施德林(P = 0.006,pseudo-F = 2.29)或安慰剂(P = 0.003,pseudo-F = 2.49, permutational multivariate analysis of variance)3 个月后,口腔微生物群的属级组成(β 多样性)有显著差异。与基线相比,使用李施德林后核分枝杆菌和副猪链球菌的数量明显增加。使用李施德林与常见口腔机会性细菌数量的增加有关,此前曾有报道称这些细菌在牙周病、食道癌、结肠直肠癌和全身性疾病中富集。这些研究结果表明,应谨慎考虑定期使用李施德林漱口水。
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