Alterations of conjunctival microbiota associated with orthokeratology lens wearing in myopic children

Study information

Reviewed Marked as Reviewed by Peace Sandy on 2024-3-26

study design

prospective cohort

Citation

PMID PubMed identifier for scientific articles.

38087200

DOI Digital object identifier for electronic documents.

https://doi.org/10.1186/s12866-023-03042-1

URI

Authors

Zhang J, Lu X, Cheng Z, Zou D, Shi W, Wang T

Journal

BMC microbiology

Year

2023

Keywords:

16S rDNA gene sequencing, Acinetobacter, Brevundimonas, Conjunctiva sac, Genomics, Microbiome, Orthokeratology lenses, Proteus

BACKGROUND: Orthokeratology (OK) lens wear increases the risk of bacterial infection, but little is known about the microbiota of the conjunctival sac in myopic children wearing OK lenses. This study aimed to investigate the changes of conjunctival microbiota in children after treatment with OK lenses using 16 S rDNA sequencing. METHODS: Twenty-eight myopic children who had been continuously wearing OK lenses for 12 to 13 months were enrolled in this prospective study. Twenty-two gender- and age-matched myopic children who had not worn OK lenses or discontinued OK lens wear at least 1 year ago were recruited as controls. Conjunctival swabs from each participant were collected for exploration of the microbiota profiles, targeting the V3-V4 regions of the 16 S rRNA gene by MiSeq sequencing. The differences in the microbial community structure and diversity were also compared between groups. RESULTS: The bacterial alpha diversity indices in the OK lens group were not different from those in the non-wearer group (P > 0.05, Wilcoxon test), while beta diversity examined using principle coordinate analysis of unweighted UniFrac divided the two groups into different clusters. Proteobacteria, Bacteroidetes, and Firmicutes were the abundant phyla in the conjunctival sac microbiota in both groups (P < 0.05, Mann-Whitney U test). Among children in the OK lens group, the Linear discriminant analysis Effect Size identified the compositional changes in OK lens-associated bacteria. Key functional genera such as Blautia, Parasutterella, and Muribaculum were enriched, whereas Brevundimonas, Acinetobacter, Proteus, and Agathobacter decreased significantly (P < 0.05, Mann-Whitney U test). Phylogenetic investigation of communities by reconstruction of unobserved states also showed altered bacterial metabolic pathways in OK lens-associated microbiota. Moreover, using receiver operating characteristic curves, Brevundimonas, Acinetobacter, Proteus, and Agathobacter alone (the area under the curve was all > 0.7500) or in combination (the area under the curve was 0.9058) were revealed to discriminate OK lens wearers from controls. CONCLUSIONS: The relative abundance of the microbial community in the conjunctival sac of myopic children can alter after OK lens wear. Brevundimonas, Acinetobacter, Proteus, and Agathobacter may be candidate biomarkers to distinguish between OK lens wearers and non-wearers.

Experiment 1

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Reviewed Marked as Reviewed by Peace Sandy on 2024-3-26

Curated date: 2024/03/08

Curator: Ebere

Revision editor(s): Ebere, Peace Sandy

Subjects

Location of subjects: China

Host species Species from which microbiome was sampled. Contact us to have more species added.: Homo sapiens

Body site Anatomical site where microbial samples were extracted from according to the Uber Anatomy Ontology: Conjunctival sac Conjunctiva serous sac,Subbrillar sac,Conjunctival sac,conjunctival sac

Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology: Myopia Close sighted,Near sighted,Near sightedness,Nearsightedness,Myopia,myopia

Group 0 name Corresponds to the control (unexposed) group for case-control studies: N group - Non-wearer group

Group 1 name Corresponds to the case (exposed) group for case-control studies: L group - Ok Lens group

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group: Myopic patients who had been wearing OK lenses for 12–13 months

Group 0 sample size Number of subjects in the control (unexposed) group: 22

Group 1 sample size Number of subjects in the case (exposed) group: 28

Antibiotics exclusion Number of days without antibiotics usage (if applicable) and other antibiotics-related criteria used to exclude participants (if any): 6 months

Lab analysis

Sequencing type: 16S

16S variable region One or more hypervariable region(s) of the bacterial 16S gene: V3-V4

Sequencing platform Manufacturer and experimental platform used for quantifying microbial abundance: Illumina

Statistical Analysis

Data transformation Data transformation applied to microbial abundance measurements prior to differential abundance testing (if any).: relative abundances

Statistical test: LEfSe

Significance threshold p-value or FDR threshold used for differential abundance testing (if any): 0.05

MHT correction Have statistical tests be corrected for multiple hypothesis testing (MHT)?: Yes

LDA Score above Threshold for the linear discriminant analysis (LDA) score for studies using the popular LEfSe tool: 3

Matched on Factors on which subjects have been matched on in a case-control study: age, sex

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness: unchanged

Chao1 Abundance-based estimator of species richness: unchanged

Simpson Estimator of species richness and species evenness: more weight on species evenness: unchanged

Richness Number of species: unchanged

Signature 1

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Reviewed Marked as Reviewed by Peace Sandy on 2024-3-26

Curated date: 2024/03/08

Curator: Ebere

Revision editor(s): Ebere, Peace Sandy

Source: Fig 4

Description: Linear Discriminant Analysis (LDA) Effect Size (LEfSe) multilevel discriminant analysis of the species differences (LDA > 3, P < 0.05, Mann–Whitney U-tests). A: Cladogram. The circle radiating from inside to outside represents the classification level from the kingdom to the genus (or species). Species with no significant differences are uniformly colored in yellow, the red nodes represent the microbial group that plays an important role in the OK group, and the green nodes represent the non-wearer group. B: Histogram of LDA value distribution shows the biomarkers with statistical difference. The LDA value represents the influence of bacterial species, and the longer the length, the higher the degree of influence. L: the OK lens group, N: the non-wearer group

Abundance in Group 1: increased abundance in L group - Ok Lens group

NCBI	Quality Control	Links
Alphaproteobacteria
Caulobacteraceae
Caulobacterales
Brevundimonas
Actinomycetota
Bacilli
Sphingomonadales
Sphingomonadaceae
Corynebacteriaceae
Mycobacteriales
Moraxellaceae
Acinetobacter
Bacillales
Staphylococcaceae
unclassified Proteus (in: enterobacteria)
Enterobacterales
Micrococcales
Staphylococcus
Proteus
Cyanobacteriota
Lysobacteraceae
Streptococcaceae
unclassified Corynebacterium
Streptococcus
Corynebacterium
Acidobacteriota
Planctomyces sp. SH-PL62
Enterococcaceae
Micrococcaceae
unclassified Enterobacter
Enterococcus
Chitinophagales
Propionibacteriales
Enterobacter
Flavobacteriales
Chitinophagaceae
Blautia
Bacillaceae
Propionibacteriaceae
Cutibacterium
Chloroflexia

Revision editor(s): Ebere, Peace Sandy

Signature 2

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Reviewed Marked as Reviewed by Peace Sandy on 2024-3-26

Curated date: 2024/03/08

Curator: Ebere

Revision editor(s): Ebere, Peace Sandy

Source: Fig 4

Description: Linear Discriminant Analysis (LDA) Effect Size (LEfSe) multilevel discriminant analysis of the species differences (LDA > 3, P < 0.05, Mann–Whitney U-tests). A: Cladogram. The circle radiating from inside to outside represents the classification level from the kingdom to the genus (or species). Species with no significant differences are uniformly colored in yellow, the red nodes represent the microbial group that plays an important role in the OK group, and the green nodes represent the non-wearer group. B: Histogram of LDA value distribution shows the biomarkers with statistical difference. The LDA value represents the influence of bacterial species, and the longer the length, the higher the degree of influence. L: the OK lens group, N: the non-wearer group

Abundance in Group 1: decreased abundance in L group - Ok Lens group

NCBI	Quality Control	Links
Bacillus
Patescibacteria group
Akkermansia sp.
Muribaculaceae
unclassified Muribaculaceae

Revision editor(s): Ebere, Peace Sandy