A comparison of sampling methods for examining the laryngeal microbiome.

Alissa S Hanshew,Susan L Thibeault,Stephanie Tadayon,Marie E Jetté,Peter E Larsen

doi:10.1371/journal.pone.0174765

Alissa S Hanshew, Susan L Thibeault + Show 3 more

Open Access

https://doi.org/10.1371/journal.pone.0174765

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Abstract

Shifts in healthy human microbial communities have now been linked to disease in numerous body sites. Noninvasive swabbing remains the sampling technique of choice in most locations; however, it is not well known if this method samples the entire community, or only those members that are easily removed from the surface. We sought to compare the communities found via swabbing and biopsied tissue in true vocal folds, a location that is difficult to sample without causing potential damage and impairment to tissue function. A secondary aim of this study was to determine if swab sampling of the false vocal folds could be used as proxy for true vocal folds. True and false vocal fold mucosal samples (swabbed and biopsied) were collected from six pigs and used for 454 pyrosequencing of the V3–V5 region of the 16S rRNA gene. Most of the alpha and beta measures of diversity were found to be significantly similar between swabbed and biopsied tissue samples. Similarly, the communities found in true and false vocal folds did not differ considerably. These results suggest that samples taken via swabs are sufficient to assess the community, and that samples taken from the false vocal folds may be used as proxies for the true vocal folds. Assessment of these techniques opens an avenue to less traumatic means to explore the role microbes play in the development of diseases of the vocal folds, and perhaps the rest of the respiratory tract.

Highlights

The role of bacterial communities in human health has been extensively researched in the past few decades, for sampled body sites such as skin [1], mouth [2], nose [3], and the intestinal tract[4]
Samples taken from true and false vocal folds were not significantly different for the three diversity metrics, the values for both Simpson and Shannon were significantly lower for Pig 3 false vocal folds versus Pig 3 true vocal folds (P
Comparative microbial community studies of the healthy and diseased human larynx are difficult to undertake due to ethical considerations in procuring vocal fold tissue

Summary

Introduction

The role of bacterial communities in human health has been extensively researched in the past few decades, for sampled body sites such as skin [1], mouth [2], nose [3], and the intestinal tract[4]. Common sampling methods include mucosal biopsy[5], brush[6], and swab[7], as well as lavage[8] and fecal collection [9]. The vocal folds are mucosal tissues housed within the larynx at the junction between the respiratory and gastrointestinal tracts and there is evidence that they have a distinct immunologic role [10] and microbial community[11]. The primary function of the vocal folds is airway protection; their vibrational properties result in voice production, making them an essential tool of human.

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