Effect of clinical parameters on the ocular surface microbiome in children and adults

Figures & data

Table 1 Patient characteristics

	Children (<18 years)	Adults (≥18 years)
Age	44 months, range	57 years, range
	5–98 months, SD 31 months	29–83 years, SD 17 years
Sex	73% male	81% male
Race	40% Hispanic	75% Caucasian
	27% Caucasian	12.5% African-American
	27% African-American	12.5% Asian
	6% not specified
Prior surgery	60% (2/3 bilateral, 1/3 unilateral)	31% (2/5 bilateral, 3/5 unilateral)
Use of topical medications	60%	69%

Figure 1 Microbial differences between adult and pediatric samples.

Notes: (A) Phylum-level annotated OTU counts (six major detected phyla) from adult and pediatric samples showing differential OTU coverage in pediatric samples, compared to adults. (B) pCoA of samples encompassing all regions and tools showing distinct clustering of adult and pediatric microbial composition (****p=0.001; Bonferroni corrected). (C) Shannon diversity index was comparable between the two groups, while adults exhibited significantly diminished (D) Chao1, (E) Faith’s PD, and (F) Observed OTU indices (test of significance – nonparametric Mann–Whitney U-test).
Abbreviations: OTU, Operational Taxonomic Unit; pCoA, principal coordinate analysis; PD, phylogenetic diversity.

Figure 2 OSM is paucibacterial in adults, but not in children.

Notes: (A) Comparison of microbiome composition between the two age groups (p=0.01 between the two age groups). (B) Same plot as “A”, color coded for different regions of sampling: Adult OSM and eyelid margin are significantly different from periocular skin but not in children, where no significant difference was found between periocular skin and eyelid margin/OSM (p=0.01 between the age groups and p=0.028 between adult periocular skin and combined eyelid margin and conjunctivae). (C) Isolated analysis of adult sample showing significantly different cluster between OSM and periocular skin (test of significance for all panels – Monte Carlo permutation with Bonferroni correction).
Abbreviations: OSM, ocular surface microbiome; pCoA, principal coordinate analysis.

Figure 3 OSM composition is comparable between the two eyes and for various clinical parameters.

Notes: (A) The right and left eye microbial composition, while distinct between age groups, showed an even distribution within each age group. (B) Similar profile was observed between the genders, (C) whether surgery was performed or not, and (D) with or without topical medication, which did not show any difference within each age group (test of significance – Monte Carlo permutation with Bonferroni correction).
Abbreviations: OSM, ocular surface microbiome; pCoA, principal coordinate analysis.

Figure S1 (A) Average 100% stacked graph of six major phyla as it compares within the adult and pediatric samples. Except Cyanobacteria, which remains unchanged between the age groups (stacked at 5%), and Fusobacteria with higher relative abundance in adults, all other phyla exhibit higher relative abundance in the pediatric samples. (B) Unweighted unifrac distance calculations for Figure 1B pCoA plot.

Notes: Distance between the groups is significantly higher compared to distance within individual groups. Test of significance – Monte Carlo permutation with Bonferroni correction. ****p<0.001.

Abbreviation: pCoA, principal coordinate analysis.

Figure S2 Phylum-wise absolute OTU counts from (A) periocular skin, (B) eyelid margin, and (C) conjunctivae. (D) Periocular skin microbiome class spread in individual adult and pediatric samples.

Notes: There is a relative expansion of Bacilli and Clostridia in adults at the expense of alpha- and beta-proteobacteria. Test of significance – nonparametric Mann–Whitney U-test.

Abbreviation: OTU, Operational Taxonomic Unit.