Lid Margin Microbiome in Stevens-Johnson Syndrome Patients With Lid Margin Keratinization and Severe Dry Eye Disease.

Purpose: The current study evaluated the lid margin microbiome of keratinized lid margins of patients with chronic Stevens-Johnson syndrome (SJS) and compared it with healthy controls and historically reported lid margin microbiome of patients with meibomian gland dysfunction (MGD). Methods: Eyelid margin swabs of 20 asymptomatic adults (mean age = 29 ± 12 years) and 10 patients with chronic SJS (mean age = 31.2 ± 14 years) with lid margin keratinization were sequenced using next generation of 16S rDNA V3 to V4 variable region. Within SJS, the keratinized lid margin microbiome was compared with adjacent eyelid skin. Results: All patients had obstructive MGD, and mean Schirmer I value was 2.8 ± 1.9 mm. The phyla were similar in two groups, whereas at the genera level, an increase in the relative abundance of Corynebacterium, Haemophilus, Azotobacter, and Afipia and a decrease of Acinetobacter was noted in SJS compared to healthy lid margins. SJS-associated microbiota displayed lesser diversity and more heterogeneity than healthy controls. The Principal Components Analysis (PCA) plot revealed wide separation in the SJS and the control groups. Correlational network analysis revealed Corynebacterium and Sphingomonas forming a major hub of negative interactions with other bacterial genera in the SJS group. Significant differences exist in the prevalent genera between keratinized lid margins and historically reported meibum microbiome of patients with MGD. In addition, the eyelid skin of patients with SJS had predominant Staphylococcus, whereas Corynebacterium and Pseudomonas were more in the keratinized lid margins compared to the eyelid skin microbiome. Conclusions: Lid margin microbiome is significantly altered in the keratinized lid margins of patients with SJS compared to the eyelid skin of patients with SJS, normal lid margins, and patients with MGD.

Inhibitory Effect of Short-Term Palpebral Margin Cleaning with Antibiotic Eye Drops on Ocular Surface Flora before Cataract Extraction: A New Preoperative Antibacterial Method.

Objective To explore the clinical significance of the combined application of palpebral margin cleaning and antibiotic eye drops in inhibiting bacterial growth in the palpebral margin and conjunctival sacs before cataract extraction. Methods In this study, 61 patients (97 eyes) with age-related cataract who underwent phacoemulsification and intraocular lens implantation were selected, and randomly grouped. In the experimental group, the combined application of palpebral margin cleaning with cotton pads and levofloxacin eye drops was given for three days before the surgery. In the control group, levofloxacin eye drops alone were applied for three consecutive days. Bacteria samples from the conjunctival sac and eyelid margins were cultivated and identified before and three days after taking antimicrobial measures, respectively. Results In the experimental group, the positive rates of the two bacteria samples were 100% (50/50) and 40% (20/50) before and 10% (5/50) and 0% (0/50) after the treatment. In the control group, the positive rates of the two bacteria samples were 97.9% (46/47) and 29.8% (14/47) before and 40.4% (19/47) and 10.6% (5/47) after the treatment. The positive rates between the two groups were not significantly different before taking antimicrobial measures (P= 0.485 and 0.395), while they were significantly different after taking antimicrobial measures (P = 0.001 and 0.024). Conclusion Combined application of eyelid and palpebral margin cleaning with cotton pads and antibiotic eye drops before cataract extraction imparted excellent antibacterial effects.

Bilateral periorbital necrotising fasciitis associated with invasive group: a Streptococcus infection.

A previously independent 56-year-old immunocompetent woman presented with septic shock in the setting of periorbital swelling and diffuse infiltrates on chest imaging. Blood cultures were positive for growth of group A Streptococcus (GAS). Broad spectrum antimicrobials were initiated with the inclusion of the antitoxin agent clindamycin. Necrosis of periorbital tissue was noted and surgical consultation was obtained. Débridement of both eyelids with skin grafting was performed. GAS was isolated from wound cultures and also observed on periorbital tissue microscopy. The final diagnosis was bilateral periorbital necrotising fasciitis (PONF) associated with invasive GAS infection. The patient had a prolonged intensive care unit course with input from multiple specialist teams. This case demonstrates the importance of early recognition and treatment of PONF, the profound systemic morbidity caused by these infections, and illustrates successful multidisciplinary teamwork.

A Genomic Approach to Investigating Ocular Surface Microorganisms: Monitoring Core Microbiota on Eyelid Margin with a Dot hybridization Assay.

A sound ocular surface microbiota has been recognized as a part of ocular surface health following a growing body of evidence from next-generation sequencing technique and metagenomic analysis. However, even from the perspective of contemporary precision medicine, it is difficult to directly apply these new technologies to clinical practice. Therefore, we proposed a model based on dot hybridization assay (DHA) to bridge conventional culture with a metagenomic approach in investigating and monitoring ocular surface microbiota. Endophthalmitis, mostly caused by bacterial infection, is the most severe complication of many intraocular surgeries, such as cataract surgery. Hazardous microorganisms hiding and proliferating in the ocular surface microbiota not only increase the risk of endophthalmitis but also jeopardize the effectiveness of the preoperative aseptic procedure and postoperative topical antibiotics. The DHA model enables the simultaneous assessment of bacterial bioburden, detection of target pathogens and microorganisms, and surveillance of methicillin/oxacillin resistance gene mecA in the ocular surface microbiota. This assay revealed heavier bacterial bioburden in men, compatible with a higher risk of endophthalmitis in male patients who underwent cataract surgery. No occurrence of endophthalmitis for these patients was compatible with non-hazardous microorganisms identified by specific dots for target pathogens. Moreover, the mecA dot detected oxacillin-resistant strains, of which culture failed to isolate. Therefore, the DHA model could provide an alternative genomic approach to investigate and monitor ocular surface microorganisms in clinical practice nowadays.

[Infection of the orbita due to Delftia acidovorans after a cat scratch]. / Orbita-Infektion mit Delftia acidovorans nach Katzenkratzer.

HISTORY: A 60 years old woman experienced a cat scratch 34 months ago on the left eyelid. Chronic, progredient skin lesions and headache developed. Treatments with cortisone, pimecrolimus, pregabalin and metamizole were not successful. After 24 months the patient complained of severe bulbus pain in the left eye, increased eye movement pain, and high photosensitivity. There were granulomatous papules in the area of the eye. FINDINGS AND DIAGNOSIS: The interdisciplinary examination findings and clinical-chemical parameters were inconspicuous. A biopsy of the eyelid area revealed the detection of Delftia acidovorans by bacterial 16S-rRNA-PCR. THERAPY AND COURSE: Treatment with piperacillin/tazobactam 3 × 4.5 g/d IV for 10 days led to rapid clinical improvement, so that the patient could be discharged after 11 days. After additional 10 months, she had no relapse and was free of complaints. CONCLUSIONS: D. acidovorans has not yet appeared as a zoonotic pathogen but should be included in the case of injury by animals in the differential diagnostic considerations.

A Case of Tinea faciei Due to Nannizzia gypsea: Inflammatory Eruption on the Medial Angle of the Eyelid.

Nannizzia gypsea is a geophilic dermatophyte, previously known as Microsporum gypseum before renaming under the new taxonomy. This organism is distributed all over the world and is considered to be involved in keratin degradation in the soil. Generally, human infection involves direct contact with fertile soil. Tinea caused by geophilic dermatophytes is much rarer than that caused by anthropophilic dermatophytes. According to the latest survey in Japan, dermatophytosis due to N. gypsea accounted for only 0.4% of cases. Clinical presentations vary and may mimic other inflammatory dermatitis, leading to incorrect diagnosis and delayed treatment. According to that past report, distal parts of the upper and lower extremities were more commonly affected, followed by the trunk, face and scalp, and rarely the nail plate. A 38-year-old woman presented with an approximately 3-week history of an itchy, solitary erythematous lesion on the left medial angle of the eyelid. Direct microscopic examination of scales revealed fungal elements, and the causative agents was identified as N. gypsea by morphological and molecular biological diagnoses. The eruption improved with systemic itraconazole treatment at 100 mg/day for 8 weeks. No recurrence has been seen for a year. However, she had no history of contact with any infectious source. Herein, we report a case of tinea faciei due to N. gypsea with an uncommon site and route of infection.

Metagenomic Profiling of Ocular Surface Microbiome Changes in Meibomian Gland Dysfunction.

Purpose: Ocular surface microbiome changes can affect meibomian gland dysfunction (MGD) development. This study aimed to delineate differences among the microbiome of eyelid skin, conjunctiva, and meibum in healthy controls (HCs) and patients afflicted with MGD. Methods: Shotgun metagenomic analysis was used to determine if there are differences between the microbial communities in ocular sites surrounding the meibomian gland in healthy individuals and patients afflicted with MGD. Results: The meibum bacterial content of these microbiomes was dissimilar in these two different types of individuals. Almost all of the most significant taxonomic changes in the meibum microbiome of individuals with MGD were also present in their eyelid skin, but not in the conjunctiva. Such site-specific microbe pattern changes accompany increases in the gene expression levels controlling carbohydrate and lipid metabolism. Most of the microbiomes in patients with MGD possess a microbe population capable of metabolizing benzoate. Pathogens known to underlie ocular infection were evident in these individuals. MGD meibum contained an abundance of Campylobacter coli, Campylobacter jejuni, and Enterococcus faecium pathogens, which were almost absent from HCs. Functional annotation indicated that in the microbiomes of MGD meibum their capability to undergo chemotaxis, display immune evasive virulence, and mediate type IV secretion was different than that in the microbiomes of meibum isolated from HCs. Conclusions: MGD meibum contains distinct microbiota whose immune evasive virulence is much stronger than that in the HCs. Profiling differences between the meibum microbiome makeup in HCs and patients with MGD characterizes changes of microbial communities associated with the disease status.

The Microbiome of the Meibum and Ocular Surface in Healthy Subjects.

Purpose: The purpose of this study was to investigate the microbiome in the meibum, conjunctival sac, and eyelid skin in young and elderly healthy subjects, and analyze the effect that age, sex, and region have on microbiome composition. Methods: This study involved 36 healthy subjects (young-age subjects: 9 men/9 women, age range: 20-35 years; elderly age subjects: 9 men/9 women, age range: 60-70 years). In all subjects, lower-eyelid meibum, lower conjunctival sac, and lower-eyelid skin specimens were collected from one eye, and then stored at -20°C. Taxonomic composition of the microbiome was obtained via 16S rRNA gene sequencing, and then analyzed. Results: The meibum microbiome showed a high α-diversity (within-community diversity), particularly in the young subjects. However, in approximately 30% of the elderly subjects, a low-diversity microbiome dominated by Corynebacterium sp. or Neisseriaceae was observed. In the young subjects, the microbiome of the meibum resembled that of the conjunctival-sac, yet in the elderly subjects, the microbiome of the conjunctival-sac became more similar to that of the eyelid skin. The eyelid-skin microbiome was relatively simple, and was typically dominated by Propionibacterium acnes in the young subjects, or by Corynebacterium sp. or Neisseriaceae in the elderly subjects. In both age groups, no significant difference was seen between the men and women in regard to the meibum, conjunctival-sac, and eyelid-skin microbiome. Conclusions: Our findings confirmed that the meibum of healthy adult-age subjects harbors highly diverse microbiota, and revealed that the meibum microbiome, especially the decrease of its diversity, alters with aging and may affect the homeostasis of the ocular surface.

Prevalence and antibiotic susceptibility of bacteria isolated in patients affected with blepharitis in a tertiary eye centre in Spain.

PURPOSE: To describe which bacteria can be found on lid margins in patients affected with blepharitis, to show their antibiotic susceptibility pattern, and to evaluate the antibiotic resistance trend of coagulase-negative Staphylococcus through time. MATERIALS AND METHODS: Consecutive cases of 198 eyes affected with blepharitis between 2012 and 2018 were reviewed. A sample was collected by rubbing a swab against the base of the eyelashes of both the eyes of all patients. The samples were inoculated in blood agar and chocolate agar. The susceptibility of the identified bacteria to common antibiotics was tested. In addition, the antibiotic susceptibility pattern of coagulase-negative Staphylococcus detected from year 2016 to 2018 was compared with that of 4 years before. RESULTS: The most common isolated bacterium was coagulase-negative Staphylococcus (89%) and Staphylococcus aureus (28%). Coagulase-negative Staphylococcus showed highest susceptibility to vancomycin (100%), neomycin (94%) and chloramphenicol (91%). Coagulase-negative Staphylococcus and Staphylococcus aureus were the most resistant to penicillin and erythromycin (resistance in 92%, 91% for coagulase-negative Staphylococcus, 86% and 43% of eyes for Staphylococcus aureus). Corynebacterium was resistant to oxacillin and erythromycin. Streptococcus viridans showed resistance to gentamycin and tobramycin. Moraxella was susceptible to most antibiotics. Bacillus was resistant to oxacillin. The antibiotic resistance trend of coagulase-negative Staphylococcus showed that the resistance to rifampicin increased through the years 2012-2018. CONCLUSION: Coagulase-negative Staphylococcus and Staphylococcus aureus were the most isolated bacteria in patients affected by blepharitis in our tertiary eye centre. Both bacteria were resistant to erythromycin. Through the years, it seems that coagulase-negative Staphylococcus gained resistance to penicillin, erythromycin, ciprofloxacin and rifampicin.

Effect of Eyelid Treatments on Bacterial Load and Lipase Activity in Relation to Contact Lens Discomfort.

OBJECTIVES: To evaluate the effect of microblepharon exfoliation on the number of eyelid bacteria and their lipase activity and the relationship of these to contact lens discomfort. METHODS: Thirty experienced contact lens wearers had their eyelid margin physiology, tear properties, and comfort scores assessed. The number, type, and frequency of lower eyelid margin bacteria, and their lipase activity, were measured. Eyelids were treated with a foam cleanser or microblepharon exfoliation. Clinical and microbiological tests were repeated at each visit. Changes and correlations were examined. RESULTS: Symptomatic lens wearers had a higher ratio for the number and frequency of gram-positive rods and cocci. Microblepharon exfoliation reduced the number and ratio of gram-positive rods to cocci from baseline for symptomatic wearers that lasted 7 to 10 days after treatment (P<0.05). Numbers of bacteria, the ratio of rods to cocci, and lipase activity correlated with lash contamination (r≥0.385; P≤0.046) and anterior blepharitis (r≥0.359; P≤0.048). Bacterial lipase correlated with meibomian gland secretions (r=0.422; P=0.038) and the tear evaporation rate (r=0.479; P=0.022). Microblepharon exfoliation produced a significant reduction in CLDEQ-8 scores and converted 10 symptomatic into asymptomatic lens wearers. CONCLUSIONS: There was dysbiosis in the lid microbiome of symptomatic lens wearers. Microblepharon exfoliation reduced the number, frequency of isolation, and ratio of gram-positive rods and cocci. Bacterial numbers and their lipase production correlated with changes to clinical signs and symptoms. Symptomatic lens wearers could be converted to asymptomatic lens wearers after microblepharon exfoliation.

The Ocular Microbiome: Molecular Characterisation of a Unique and Low Microbial Environment.

Aim: The ocular surface is continually exposed to bacteria from the environment and traditional culture-based microbiological studies have isolated a low diversity of microorganisms from this region. The use of culture-independent methods to define the ocular microbiome, primarily involving 16S ribosomal RNA gene sequencing studies, have shown that the microbial communities present on the ocular surface have a greater diversity than previously reported. Method: A review of the literature on ocular microbiome research in health and disease. Results: Molecular techniques have been used to investigate the effect of contact lens wear and disease on the microbiota of the ocular surface and eyelids and the immunoregulatory role of the ocular surface microbiota. Studies have shown that compositional changes in the microbiota occur in ocular surface disorders such as blepharitis, trachoma and dry eye and also suggest a role of the ocular and non-ocular microbiome in retinal disease including age-related macular degeneration, glaucoma, uveitis and diabetic retinopathy. However, ocular microbiome studies need to recognise the potential for contamination to impact findings and carefully control each stage of the experimental procedure and to utilise statistical methods to identify contamination signals. Conclusion: The healthy ocular surface is characterised by a relatively stable, comparatively low diversity microbiome with recent findings that the bacteria of the ocular surface appear to have a role in maintaining homeostasis by modulating immune function.

Biogeography of the human ocular microbiota.

PURPOSE: The human eye is composed of numerous microhabitats. The aim of this study was to understand the communality and differences in the microbiomes of various regions of the eye. METHODS: Four ocular sites from different subject groups were assessed including the eyelid margin tissue from patients with lid abnormalities (n = 20), fornix and limbus conjunctival tissue from patients with pterygia (n = 23), ocular (conjunctival) surface swabs (n = 45) and facial skin swabs (n = 16). Microbial communities were analysed by extracting total DNA from samples and sequencing the 16S ribosomal(r)RNA gene using the Illumina MiSeq platform. Sequences were quality filtered, clustered into unique sequences (zOTUs) using the UNOISE pipeline in USEARCH and taxonomically classified using SILVA. RESULTS: A difference in bacterial richness and diversity was found between sites (P < 0.001) and for age (P < 0.035) but not for sex (P > 0.05). There was a difference in bacterial community structure and composition between sites (P < 0.001). Bacterial distribution could be broadly classified into three groups - zOTUs resident on the skin and lid margin but with low abundances at other sites (Corynebacterium, Staphylococcus), zOTUs found mainly on the ocular surface (Acinetobacter, Aeribacillus) and zOTUs mostly present in the conjunctiva and lid margin (Pseudomonas). CONCLUSION: The microhabitats of the human eye (ocular surface, conjunctiva, lid margin and skin) have a distinct bacterial biogeography with some bacteria shared between multiple regions while other bacteria occupy a more confined niche.

Orbital tuberculosis: perspectives from Victoria, Australia.

PURPOSE: Orbital tuberculosis (TB) is a rare extra-pulmonary manifestation of tuberculosis and its clinical diagnosis poses unique challenges, with potential for destructive complications as well as social and public health implications. The aim of this study is to report our experience of patients presenting with orbital TB and to identify common aspects. METHODS: A systematic search for mandatory notifications of orbital tuberculosis between January 01, 1994 and December 12, 2016 was undertaken in the Victorian Tuberculosis database. In addition, members of the Australian and New Zealand Society of Ophthalmic Plastic Surgeons (ANZSOPS) were surveyed to identify cases of orbital tuberculosis diagnosed on biopsy in the past 20 years. Medical case notes of identified cases were reviewed retrospectively. RESULTS: Three cases were identified as having occurred in Victoria, aged 44-59 years old. All cases had emigrated from endemic countries with higher tuberculosis burden. Diagnosis of tuberculosis was often difficult due to few or non-viable acid fast bacilli and low yield of positive culture in paucicellular orbital specimens. CONCLUSIONS: Orbital TB is rare but remains an important differential diagnosis of orbital mass lesions. The diagnosis of orbital TB requires a high index of clinical suspicion and targeted investigations in patients originating from endemic areas. Diagnosis and treatment rely on effective collaboration between ophthalmologists, infectious disease physicians, and pathologists.

Effect of Eyelid Margin Cleaning on Conjunctival and Eyelid Bacterial Flora Before Intraocular Surgery.

PURPOSE: To compare the bacterial cultures of the lower eyelid margins and lower fornices between ocular irrigations with and without eyelid margin cleaning before intraocular surgery. METHODS: A prospective study was carried out that included 102 eyes from 51 patients undergoing intraocular surgery between December 2016 and April 2017. In each patient, the surgical eye (control group) only received ocular irrigation and the opposite eye (study group) received ocular irrigation with eyelid margin cleaning. Normal saline solution was used in all procedures. In each group, samples from the lower eyelid margins and lower fornices were cultured before and after cleaning. Povidone iodine and antibiotic eye drops were instilled after specimen collection. RESULTS: The most common bacterial isolate of all groups was Staphylococcus spp. There were no statistically significant differences of positive cultures between the control and study groups in either lower eyelid margins (Odds ratio [OR], 2.19; 95% CI, 0.84-5.72) nor lower fornices (OR, 0.93; 95% CI, 0.43-2.00). In addition, there were no statistically significant differences of positive cultures between before and after eyelid cleanings from both lower eyelid margins (OR, 2.74; 95% CI, 0.53-14.24) and lower fornices (OR, 0.73; 95% CI, 0.21-2.57). CONCLUSIONS: Eyelid margin cleaning with normal saline solution immediately before intraocular surgery did not help to decrease microbial loading on the ocular surfaces. The rate of bacterial contamination, however, was not increased which could be due to not performing eyelid compression.

Bullous eyelid.

Periocular necrotising fasciitis is a rare but sight-threatening condition, which relies on clinical judgement to detect in a timely manner. A 51-year-old woman presented to a rural hospital with rapid onset bilateral eye swelling, erythema and pain and was started on broad spectrum intravenous antibiotics. Upon admission, she became septic and required fluid resuscitation and transfer to a higher level of care. She received debridement and continued intravenous antibiotics, with step down to oral when clinically stable. Rapid recognition and treatment of her condition resulted in a positive outcome.

Periocular necrotizing fasciitis in an infant.

Periocular necrotizing fasciitis developed in a 12-month-old boy with swelling of both eyes and redness and a discharge from the left eye approximately 36 hours after blunt trauma. Computed tomography revealed preseptal and soft-tissue edema on the left side, but no signs of orbital involvement, orbital fractures, or drainable abscess in the anterior left lower eyelid. The inflammatory signs worsened over the next day, and there was purulent discharge from the left lower eyelid and an abscess and necrosis of the lower eyelid skin. He did well following surgical debridement and treatment with intravenous antibiotics. His course highlights the difficulty in diagnosing necrotizing fasciitis and the necessity for prompt surgical debridement and empirical broad-spectrum antibiotic therapy.

Baby Shampoo Versus Povidone-Iodine or Isopropyl Alcohol in Reducing Eyelid Skin Bacterial Load.

PURPOSE: Baby shampoo is used as an alternative surgical skin preparation, but the evidence supporting its use is scarce with no descriptions of efficacy in the periocular region. The authors compare the efficacy of baby shampoo, povidone-iodine (PI, Betadine) and isopropyl alcohol (IA) in reducing eyelid skin bacterial load. METHODS: Prospective, randomized, comparative, and interventional trial. Bacterial load on adult, human eyelid skin was quantitated before and after cleansing with 1) dilute baby shampoo, 2) 10% PI, or 3) 70% IA. Paired skin swabs were collected from a 1 cm area of the upper eyelid of subjects before and after a standardized surgical scrub technique. Samples were cultured on 5% sheep blood agar for 24 hours. The number of colony forming units (CFU) was assessed and bacterial load per square centimeter of eyelid skin was quantified. RESULTS: Baseline and postcleansing samples were assessed from 42 eyelids of 42 subjects (n = 14 for each of baby shampoo, PI, and IA). Before cleansing, similar amounts of bacterial flora were grown from all specimens (median log CFU/cm = 2.04 before baby shampoo, 2.01 before PI, 2.11 before IA; p > 0.05). All 3 cleansing agents significantly reduced the bacterial load (p < 0.01 for each). There was no statistically significant difference in postcleansing bacterial load between the 3 cleansing agents (median log CFU/cm = 0.48 after baby shampoo, 0.39 after PI, 0.59 after IA; p > 0.05). Change from baseline in bacterial load was statistically similar for all 3 agents (median reduction in log CFU/cm = 1.28 with baby shampoo, 1.57 with PI, 1.40 with IA; p > 0.05). These corresponded to bacterial load reductions of 96.3%, 96.6%, and 98.4% for baby shampoo, PI, and IA, respectively. CONCLUSIONS: Baby shampoo achieved comparable diminution in eyelid skin bacterial load to PI or IA. These data suggest baby shampoo may be an effective preoperative cleansing agent.

Temporal Stability and Composition of the Ocular Surface Microbiome.

To determine if there is a core ocular surface microbiome and whether there are microbial community changes over time, the conjunctiva of 45 healthy subjects were sampled at three time points over three months and processed using culture-dependent and -independent methods. Contaminant taxa were removed using a linear regression model using taxa abundances in negative controls as predictor of taxa abundances in subject samples. Both cultured cell counts and sequencing indicated low microbial biomass on the ocular surface. No cultured species was found in all subjects at all times or in all subjects at any one time. After removal of contaminant taxa identified in negative controls using a statistical model, the most commonly detected taxon was Corynebacterium (11.1%). No taxa were found in all subjects at all times or in all subjects in any one time, but there were 26 taxa present in at least one or more subjects at all times including Corynebacterium and Streptococcus. The ocular surface contains a low diversity of microorganisms. Using culture dependent and independent methods, the ocular surface does not appear to support a substantial core microbiome. However, consistently present taxa could be observed within individuals suggesting the possibility of individual-specific core microbiomes.