Alterations in the conjunctival surface bacterial microbiome in bacterial keratitis patients

Shivaji, Sisinthy; Jayasudha, Rajagopalaboopathi; Chakravarthy, Sama Kalyana; SaiAbhilash, Chinthala Reddy; Prashanthi, Gumpili Sai; Sharma, Savitri; Garg, Prashant; Murthy, Somasheila I

doi:10.1016/j.exer.2020.108418

Cited by 28 publications

(25 citation statements)

References 82 publications

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“… Shivaji et al (2021) recently published similar study. Some of their patients had received recently antibiotic treatment, and a few healthy controls had undergone photorefractive keratectomy.…”

Section: Discussionmentioning

confidence: 57%

“…When the network relationship is thoroughly deciphered, the prevention and treatment of oculopathy through regulation of ocular microbiota becomes promising. Shivaji et al (2021) recently published similar study. Some of their patients had received recently antibiotic treatment, and a few healthy controls had undergone photorefractive keratectomy.…”

Section: The Imbalance Of Protective and Aggressive Bacteria In The Ocular Microbiota For Healthy People Triggers Susceptibility To Bkmentioning

confidence: 57%

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Profiling of Diagnostic Information of and Latent Susceptibility to Bacterial Keratitis From the Perspective of Ocular Bacterial Microbiota

Ren

Liu

et al. 2021

Front. Cell. Infect. Microbiol.

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The ocular surface possesses its own bacterial microbiota. Once given a chance, opportunistic pathogens within ocular microbiota may lead to corneal infection like bacterial keratitis (BK). To reveal the possible factor that makes people vulnerable to BK from the perspective of ocular bacterial microbiota, as well as to compare diagnostic information provided by high-throughput 16S rDNA sequencing and bacterial culture, 20 patients with BK and 42 healthy volunteers were included. Conjunctival swabs and corneal scrapings collected from the diseased eyes of BK patients were subjected for both high-throughput 16S rDNA sequencing and bacterial culture. Conjunctival swabs collected from the normal eyes of BK patients and healthy volunteers were sent only for sequencing. For identifying the pathogens causing BK, high-throughput 16S rDNA sequencing presented a higher positive rate than bacterial culture (98.04% vs. 17.50%), with 92.11% reaching the genus level (including 10.53% down to the species level). However, none of the sequencing results was consistent with the cultural results. The sequencing technique appears to challenge culture, and could be a complement for pathogen identification. Moreover, compared to the eyes of healthy subjects, the ocular microbiota of three sample groups from BK patients contained significantly less Actinobacteria and Corynebacteria (determinate beneficial symbiotic bacteria), but significantly more Gammaproteobacteria, Pseudomonas, Bacteroides, and Escherichia-Shigella (common ocular pathogenic bacteria). Therefore, it is speculated that the imbalance of protective and aggressive bacteria in the ocular microbiota of healthy people may trigger susceptibility to BK. Based on this speculation, it seems promising to prevent and treat infectious oculopathy through regulating ocular microbiota.

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Section: Discussionmentioning

confidence: 57%

Section: The Imbalance Of Protective and Aggressive Bacteria In The Ocular Microbiota For Healthy People Triggers Susceptibility To Bkmentioning

confidence: 57%

Profiling of Diagnostic Information of and Latent Susceptibility to Bacterial Keratitis From the Perspective of Ocular Bacterial Microbiota

Ren

Liu

et al. 2021

Front. Cell. Infect. Microbiol.

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“…to eliminate pathogens [ 5 , 6 , 7 ]. Despite these defense mechanisms, many microorganisms do survive on the ocular surface and recent studies either based on 16S ribosomal RNA (rRNA) gene amplification, cloning and sequencing or by using NGS (next-generation sequencing) based on 16S rRNA gene amplification and analysis (16S rRNA meta-barcoding) revealed a greater degree of diversity and abundance in the bacterial microbiome of the ocular surface [ 8 , 9 , 10 , 11 , 12 , 13 ]. Keilty [ 14 ] was the first to cultivate hemolytic Staphylococcus from the conjunctival swabs of normal subjects.…”

Section: Introductionmentioning

confidence: 99%

Biofilm-Forming Potential of Ocular Fluid Staphylococcus aureus and Staphylococcus epidermidis on Ex Vivo Human Corneas from Attachment to Dispersal Phase

2021

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The biofilm-forming potential of Staphylococcus aureus and Staphylococcus epidermidis, isolated from patients with Endophthalmitis, was monitored using glass cover slips and cadaveric corneas as substrata. Both the ocular fluid isolates exhibited biofilm-forming potential by the Congo red agar, Crystal violet and 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-(phenylamino) carbonyl-2H-tetra-zolium hydroxide (XTT) methods. Confocal microscopy demonstrated that the thickness of the biofilm increased from 4–120 h of biofilm formation. Scanning electron microscopic studies indicated that the biofilms grown on cover slips and ex vivo corneas of both the isolates go through an adhesion phase at 4 h followed by multilayer clumping of cells with intercellular connections and copious amounts of extracellular polymeric substance. Clumps subsequently formed columns and eventually single cells were visible indicative of dispersal phase. Biofilm formation was more rapid when the cornea was used as a substratum. In the biofilms grown on corneas, clumping of cells, formation of 3D structures and final appearance of single cells indicative of dispersal phase occurred by 48 h compared to 96–120 h when biofilms were grown on cover slips. In the biofilm phase, both were several-fold more resistant to antibiotics compared to planktonic cells. This is the first study on biofilm forming potential of ocular fluid S. aureus and S. epidermidis on cadaveric cornea, from attachment to dispersal phase of biofilm formation.

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“…Amplification of the V3–V4 region of bacterial 16S rRNA genes was performed as described earlier. 13 Bacterial microbiomes were generated following the standard Illumina (San Diego, CA, USA) protocol 13 , 25 and sequenced using 2 × 250-bp chemistry on an Illumina MiSeq system with a paired-end protocol at Xcelris Genomics Pvt. Ltd. (Ahmedabad, India).…”

Section: Methodsmentioning

confidence: 99%

“…SILVA 138 SSU ( https://www.arb-silva.de/documentation/release-138/ ) was used as the reference OTU database to identify bacteria at 97% identity, and the assignment of taxonomy was made as described in our earlier studies. 17 , 25 Sparse OTUs (<0.001% of the total HQ reads) were not considered.…”

Section: Methodsmentioning

confidence: 99%

Dysbiosis in the Gut Microbiome in Streptozotocin-Induced Diabetes Rats and Follow-Up During Retinal Changes

Padakandla

Das

Prashanthi

et al. 2021

Invest. Ophthalmol. Vis. Sci.

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Purpose To analyze the gut bacterial microbiome of streptozotocin-induced diabetic rats and rats with retinal changes. Methods Induction of diabetes was confirmed by an increase in blood sugar (>150 mg/dL), and the progression of diabetes with retinal changes was assessed by histology and immunohistochemistry of retinal sections. Microbiomes were generated using fecal DNA, and the V3–V4 amplicons were sequenced and analyzed by QIIME and R. Results Dysbiosis in the gut microbiome of diabetic rats and diabetic rats with retinal changes was observed at the phylum and genus levels compared with the control rats. Heat-map analysis based on the differentially abundant genera indicated that the microbiomes of controls and diabetic rats separated into two distinct clusters. The majority of the microbiomes in diabetic rats with retinal changes also formed a distinct cluster from the control rats. β-diversity analysis separated the microbiome of control rats from the microbiome of diabetic rats and diabetic rats with retinal changes, but the microbiomes of diabetic rats and diabetic rats with retinal changes showed an overlap. Functional analysis indicated that the enhanced inflammation in diabetic rats showing retinal changes could be ascribed to a decrease in anti-inflammatory bacteria and an increase in pathogenic and proinflammatory bacteria. Conclusions This study showed that the gut bacterial microbiome in diabetic rats with retinal changes was different compared with control rats. The results could help develop novel therapeutics for diabetics and diabetic individuals with retinal changes.

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Alterations in the conjunctival surface bacterial microbiome in bacterial keratitis patients

Cited by 28 publications

References 82 publications

Profiling of Diagnostic Information of and Latent Susceptibility to Bacterial Keratitis From the Perspective of Ocular Bacterial Microbiota

Profiling of Diagnostic Information of and Latent Susceptibility to Bacterial Keratitis From the Perspective of Ocular Bacterial Microbiota

Biofilm-Forming Potential of Ocular Fluid Staphylococcus aureus and Staphylococcus epidermidis on Ex Vivo Human Corneas from Attachment to Dispersal Phase

Dysbiosis in the Gut Microbiome in Streptozotocin-Induced Diabetes Rats and Follow-Up During Retinal Changes

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