Ocular surface microbiota in patients with aqueous tear-deficient dry eye

From BugSigDB
Reviewed Marked as Reviewed by Folakunmi on 2024-1-5
study design
Citation
PMID PubMed identifier for scientific articles.
DOI Digital object identifier for electronic documents.
URI
Authors
Andersson J, Vogt JK, Dalgaard MD, Pedersen O, Holmgaard K, Heegaard S
Journal
The ocular surface
Year
2021
Keywords:
16S rRNA gene amplicon sequencing, ADDE, Bacilli class, Bacterial biomarker, Dry eye, Ocular surface microbiota
PURPOSE: An altered ocular surface microbiota may contribute to the pathophysiology of dry eye disease. The aim of the study was to explore potential differences in microbiota diversity and composition in aqueous tear-deficient dry eye (with and without ocular graft-versus-host disease) compared with controls. METHODS: Swab samples from the inferior fornix of the conjunctiva were obtained from patients with aqueous tear-deficient dry eye with and without ocular graft-versus-host disease (n = 18, n = 21, respectively) and controls (n = 28). Isolated bacterial DNA from swabs were analyzed with 16S rRNA gene amplicon sequencing. RESULTS: Decreased microbiota diversity was observed in patients with aqueous tear-deficient dry eye (p ≤ 0.003) who also showed a difference in microbiota composition compared with controls (p = 0.001). Although several genera were less abundant in aqueous tear-deficient dry eye, a minimal core ocular surface microbiota comprising five genera was shared by >75% of the study participants: Enhydrobacter, Brevibacterium, Staphylococcus, Streptococcus and Cutibacterium. Pseudomonas was identified as a bacterial biomarker for controls and Bacilli for patients with aqueous tear-deficient dry eye. CONCLUSIONS: Ocular surface microbiota in patients with aqueous tear-deficient dry eye was characterized by an aberrant microbiota composition in comparison to controls, with decreased diversity and reduced relative abundances of several genera. Additionally, a few genera were present in most of the study population, indicating that a minimal core ocular surface microbiota may exist.

Experiment 1


Reviewed Marked as Reviewed by Folakunmi on 2024-1-5

Curated date: 2023/09/04

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Folakunmi

Subjects

Location of subjects
Denmark
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 fornix Conjunctiva fornix,Forniceal conjunctiva,Fornix conjunctiva,Conjunctival fornix,conjunctival fornix
Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology
Dry eye syndrome dry eye,Dry Eye Syndrome,dry eye syndrome,Dry Eye Syndromes,dry eye(s),eye(s), dry,KCS,Keraconjunctivitis sicca,Keratoconjunctivitis Sicca,Keratoconjunctivitis sicca,keratoconjunctivitis sicca,Keratoconjunctivitis sicca (disorder),sicca, keratoconjunctivitis,Tear film insufficiency,tear film insufficiency,Dry eye syndrome
Group 0 name Corresponds to the control (unexposed) group for case-control studies
control
Group 1 name Corresponds to the case (exposed) group for case-control studies
Dry Eye
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
Patients with Aqueous Deficient Dry Eye without Ocular Graft vs Host Disease
Group 0 sample size Number of subjects in the control (unexposed) group
28
Group 1 sample size Number of subjects in the case (exposed) group
21
Antibiotics exclusion Number of days without antibiotics usage (if applicable) and other antibiotics-related criteria used to exclude participants (if any)
For controls- those with hx of recent (within 3 mos) use of antibiotics (local or systemic) were excluded. ADDE patients were allowed and 7/21 had received local or systemic antibiotics within the previous 3 mos.

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).
raw counts
Statistical test
DESeq2
Significance threshold p-value or FDR threshold used for differential abundance testing (if any)
0.05

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness
decreased
Richness Number of species
decreased

Signature 1

Reviewed Marked as Reviewed by Folakunmi on 2024-1-5

Curated date: 2023/09/04

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Folakunmi

Source: Figure 4a, within result text (Several genera were less abundant in ADDE, lines 8-15)

Description: Comparison of the relative abundances of genera with DESeq2 between patients with DryEye and Control

Abundance in Group 1: decreased abundance in Dry Eye

NCBI Quality ControlLinks
Corynebacterium
Flavobacterium
Fusobacterium
Haemophilus
Massilia
Microbacterium
Ottowia
Rothia
Sphingomonas
Thermus
Veillonella

Revision editor(s): Mary Bearkland, Folakunmi

Experiment 2


Reviewed Marked as Reviewed by Folakunmi on 2024-1-5

Curated date: 2023/09/04

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Folakunmi

Differences from previous experiment shown

Subjects

Group 1 name Corresponds to the case (exposed) group for case-control studies
OGVHD
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
Patients with Aqueous Deficient Dry Eye and Ocular Graft vs Host Disease as diagnosed using the criteria described by the International Chronic Ocular Graft-vs-Host-Disease (GVHD) Consensus. Group and the patients included were classified as either probable or definite chronic GVHD
Group 1 sample size Number of subjects in the case (exposed) group
18

Lab analysis

Statistical Analysis

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness
decreased
Richness Number of species
decreased

Signature 1

Reviewed Marked as Reviewed by Folakunmi on 2024-1-5

Curated date: 2023/09/04

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Folakunmi

Source: Figure 4b, within result text (Several genera were less abundant in ADDE, line 19)

Description: Comparison of the relative abundances of genera with DESeq2 between patients with OGVHD and Control

Abundance in Group 1: decreased abundance in OGVHD

NCBI Quality ControlLinks
Carnobacterium
Cloacibacterium
Listeria
Microbacterium
Pseudomonas

Revision editor(s): Mary Bearkland, Folakunmi

Signature 2

Reviewed Marked as Reviewed by Folakunmi on 2024-1-5

Curated date: 2023/09/04

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Folakunmi

Source: Figure 4b, within result text (Several genera were less abundant in ADDE, line 20)

Description: Comparison of the relative abundances of genera with DESeq2 between patients with OGVHD and Control

Abundance in Group 1: increased abundance in OGVHD

NCBI Quality ControlLinks
Bacillus

Revision editor(s): Mary Bearkland, Folakunmi

Experiment 3


Reviewed Marked as Reviewed by Folakunmi on 2024-1-5

Curated date: 2023/09/04

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland

Differences from previous experiment shown

Subjects

Group 1 name Corresponds to the case (exposed) group for case-control studies
ADDE
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
Patients with Aqueous Deficient Dry Eye including those with and without Ocular Graft vs Host Disease as diagnosed using the criteria described by the International Chronic Ocular Graft-vs-Host-Disease (GVHD) Consensus. Group and the patients included were classified as either probable or definite chronic GVHD
Group 1 sample size Number of subjects in the case (exposed) group
39

Lab analysis

Statistical Analysis

Data transformation Data transformation applied to microbial abundance measurements prior to differential abundance testing (if any).
relative abundances
Statistical test
LEfSe
LDA Score above Threshold for the linear discriminant analysis (LDA) score for studies using the popular LEfSe tool
3


Signature 1

Reviewed Marked as Reviewed by Folakunmi on 2024-1-5

Curated date: 2023/09/04

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland

Source: Figure 5

Description: Fig. 5. Bacterial biomarkers identified with the LEfSe algorithm. A. Circular cladogram displaying the relation between taxa at different taxonomic levels (i.e. phylum to genus level). Among the enriched bacteria (biomarkers) in aqueous tear-deficient dry eye (ADDE) (red) was Bacilli and for controls (green) was Pseudomonadaceae. The size of the nodes represents the taxa abundance. B. Scores for linear discriminant analysis (LDA) with LEfSe tool for taxa with LDA score >3 and p <0.05 are shown in the histogram. Similarly, as the cladogram demonstrated, among the bacterial biomarkers, Pseudomonas was identified for controls and Bacilli for ADDE.

Abundance in Group 1: increased abundance in ADDE

NCBI Quality ControlLinks
Bacilli
Bacillota
Brevibacterium
Enhydrobacter
Moraxellaceae
Brevibacteriaceae

Revision editor(s): Mary Bearkland

Signature 2

Reviewed Marked as Reviewed by Folakunmi on 2024-1-5

Curated date: 2023/09/04

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland

Source: Figure 5

Description: Fig. 5. Bacterial biomarkers identified with the LEfSe algorithm. A. Circular cladogram displaying the relation between taxa at different taxonomic levels (i.e. phylum to genus level). Among the enriched bacteria (biomarkers) in aqueous tear-deficient dry eye (ADDE) (red) was Bacilli and for controls (green) was Pseudomonadaceae. The size of the nodes represents the taxa abundance. B. Scores for linear discriminant analysis (LDA) with LEfSe tool for taxa with LDA score >3 and p <0.05 are shown in the histogram. Similarly, as the cladogram demonstrated, among the bacterial biomarkers, Pseudomonas was identified for controls and Bacilli for ADDE.

Abundance in Group 1: decreased abundance in ADDE

NCBI Quality ControlLinks
Carnobacteriaceae
Carnobacterium
Flavobacteriaceae
Pseudomonadaceae
Pseudomonadota
Pseudomonas
Mycobacteriales
Corynebacteriaceae
Micrococcaceae

Revision editor(s): Mary Bearkland