Altered Eye Microbiota by Wearing Contact Lenses A Non-lens wearers Time points O Conjunctiva O skin under the eye争 3o8:中 Pc1(12% B Non-lens wearers vs Lens wearers D g farers) Non-lens wearer Conjunctiva ]: n-s wEarers Non-ens wearers Lens wearers FIG 2 Beta diversity in conjunctiva, skin, and contact lens samples between non-lens wearers and lens wearers. Unweighted Unifrac distances were used to evaluate diversities between samples. (A)PCoA plot of bacterial communities by body site, time, and subject in non-lens wearers(left)and lens wearers(right) PCI Box plots of intragroup and intergroup di in the ocular microbiota of non-lens wearers and lens wearers. *+, Pvalue= <0.001 However, regardless of wearing or not wearing lenses, Source O Conjunctiva(Non-lens wearers)(n=64) Tracker analyses using the v4 region from data generated in a O Conjunctiva( Lens wearers)(n=52) previous study of the skin microbiome(16)showed that the con- Skin under the eye(n=114) junctival microbiota of both lens wearers and non-lens wearers had higher proportions of hand -like bacteria than of face-like bac teria(P<00001; see Fig. S3B in the supplemental material) O Skin HMP(n=2, 199 There were no significant differences in bacterial diversity and O Stool HMP(n=437) composition between the conjunctival microbiota and skin mi- O Vaginal HMP(n=564) crobiota at different time points(see Fig. S6 in the supplemental material) Predictive functional profiling showed that the conjunctiva of lens wearers had a higher proportion of bacterial genes related to ell motility and xenobiotic biodegradation than the conjunctiva of non-lens wearers. On the other hand, the conjunctival micro- biota of non-wearers had higher relative content of genes related to genetic processing and nucleotide metabolism(see Fig S7 in lemental material DISCUSSION Wearing contact lenses is a known risk factor for development of microbial keratitis and other inflammatory eye conditions(17, 18). Several researchers have tried to identify the causative organ- PC1(16%)ism of these contact lens-related diseases using culture-dependent methods(1, 18). To the best of our knowledge, this was a pioneer n, and contact lens samples with ing study in identifying the structure of the microbiota from con- weighted Un ots (green, purple, cyan, and pink) indi- tact lenses and conjunctiva of contact lenses wearers using 16S and large dots(red, orange, yellow, and rRNA gene sequencing. While mostly illustrated by results of blue) indicate studies of the gut, the concept of colonization resistance, a protec March/April 2016 Volume 7 Issue 2 e00198-16 msio° mbio. asm. org3However, regardless of wearing or not wearing lenses, Source￾Tracker analyses using the V4 region from data generated in a previous study of the skin microbiome (16) showed that the con￾junctival microbiota of both lens wearers and non-lens wearers had higher proportions of hand-like bacteria than of face-like bac￾teria (P 0.0001; see Fig. S3B in the supplemental material). There were no significant differences in bacterial diversity and composition between the conjunctival microbiota and skin mi￾crobiota at different time points (see Fig. S6 in the supplemental material). Predictive functional profiling showed that the conjunctiva of lens wearers had a higher proportion of bacterial genes related to cell motility and xenobiotic biodegradation than the conjunctiva of non-lens wearers. On the other hand, the conjunctival micro￾biota of non-wearers had higher relative content of genes related to genetic processing and nucleotide metabolism (see Fig. S7 in the supplemental material). DISCUSSION Wearing contact lenses is a known risk factor for development of microbial keratitis and other inflammatory eye conditions (17, 18). Several researchers have tried to identify the causative organ￾ism of these contact lens-related diseases using culture-dependent methods (1, 18). To the best of our knowledge, this was a pioneer￾ing study in identifying the structure of the microbiota from con￾tact lenses and conjunctiva of contact lenses wearers using 16S rRNA gene sequencing. While mostly illustrated by results of studies of the gut, the concept of colonization resistance, a protec￾FIG 2 Beta diversity in conjunctiva, skin, and contact lens samples between non-lens wearers and lens wearers. Unweighted UniFrac distances were used to evaluate diversities between samples. (A) PCoA plot of bacterial communities by body site, time, and subject in non-lens wearers (left) and lens wearers (right). PC1, first principal component; PC2, second principal component. (B) Box plots of intragroup and intergroup distances in bacterial communities between conjunctiva and skin under the eye. (C) PCoA plot of ocular microbiota in non-lens wearers and lens wearers with bacterial communities of contact lenses. (D) Box plots of intragroup and intergroup distances in the ocular microbiota of non-lens wearers and lens wearers. ***, P value 0.001. FIG 3 Beta diversity in conjunctiva, skin, and contact lens samples with samples from the HMP database. A PCoA plot was generated using un￾weighted UniFrac distances. Small dots (green, purple, cyan, and pink) indi￾cate samples from the HMP database, and large dots (red, orange, yellow, and blue) indicate samples from this study. 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