Levofloxacin susceptibility of Staphylococci from conjunctiva in patients with atopic dermatitis.

PURPOSE: The ocular surface in patients with atopic dermatitis (AD) is known to harbor an abundance of gram-positive cocci, particularly Staphylococcus aureus (S. aureus). This study reviewed the results of microbial cultures from the conjunctiva in AD patients, with special attention to the levofloxacin susceptibility of Staphylococci. STUDY DESIGN: Retrospective, single-center study. METHODS: This study involved 131 eyes of 112 Japanese patients with AD (87 men and 25 women, mean age: 40.4 ± 12.2 years) who underwent ocular surgery at Kyorin University Hospital. Bacterial isolates were collected from the conjunctival sacs in the preoperative period. Drug resistance to methicillin and levofloxacin was judged using the minimal inhibitory concentrations of oxacillin and levofloxacin determined by the broth dilution method. RESULTS: One hundred and fifty-seven strains were identified in 103 of the 131 eyes examined. S. aureus was isolated from 74 eyes (56.5%), followed by Staphylococcus epidermidis (S. epidermidis). In S. aureus, 11 strains (14.9%) were methicillin-resistant, and 18 (24.3%) were levofloxacin-resistant. In S. epidermidis, 15 strains (26.8%) were methicillin-resistant, and 17 (30.4%) were levofloxacin-resistant. No significant differences were observed in levofloxacin susceptibility with age, sex, previous ocular surgery, or duration of previous surgery. However, logistic multivariate analysis revealed that levofloxacin-resistant Staphylococci were concurrently resistant to methicillin, suggesting multidrug resistance. CONCLUSION: Distinctive bacterial distribution and drug resistance need consideration in the managing of ocular disorders among patients with AD.

The Conjunctival Microbiome and Dry Eye: What We Know and Controversies.

ABSTRACT: Dry eye disease is a common multifactorial condition that may be idiopathic or associated with autoimmune conditions, such as Sjogren syndrome. Commensal microorganisms modify immune responses, so it is relevant to understand how they modify such immune-mediated diseases. Microbiota in the gut regulate inflammation in the eye, and conversely, severe inflammation of the ocular surface results in alteration of gut microbiome. The conjunctiva microbiome can be analyzed using 16S or shotgun metagenomics. The amount of microbial DNA in ocular surface mucosa relative to human DNA is limited compared with the case of the intestinal microbiome. There are challenges in defining, harvesting, processing, and analyzing the microbiome in the ocular surface mucosa. Recent studies have shown that the conjunctiva microbiome depends on age, presence of local and systemic inflammation, and environmental factors. Microbiome-based therapy, such as the use of oral probiotics to manage dry eye disease, has initial promising results. Further longitudinal studies are required to investigate the alteration of the conjunctival microbiome after local therapy and surgery.

Topical Antiseptics in Minimizing Ocular Surface Bacterial Load Before Ophthalmic Surgery: A Randomized Controlled Trial.

PURPOSE: To investigate the reduction of the ocular surface bacterial load induced by 2 commercially available ophthalmic antiseptic formulations, povidone-iodine (PVI) 0.6% and chlorhexidine (CLX) 0.02%, before ocular surgery. DESIGN: Randomized controlled trial. METHODS: Seventy adult patients undergoing intraocular surgery (phacoemulsification) were randomized to receive in the index eye PVI (group A) 4 times a day for 3 days or CLX (group B) 4 times a day for 3 days before surgery. The untreated eye was used as control. A conjunctival swab was taken in both eyes before (T0) and after (T1) therapy. Microbial DNA was quantified with real-time polymerase chain reaction (PCR) analysis. The Mick algorithm was used to compare the abundance of each genus/genera against the distribution of abundances from the reference. At T1, patients filled a questionnaire to evaluate therapy-induced symptoms. Primary outcome was the reduction of bacterial DNA at T1 (microbial load), vs control arm, expressed as mean number of real-time PCR cycle times (CTs). Secondary outcomes were taxonomic composition, differential abundance, and therapy-induced ocular symptoms. RESULTS: The T0-T1 difference in CT was significant in group B, but not in group A (mean [95% CI], 0.99 [0.33] vs 0.26 [0.15], P < .001, and 0.65 [0.3] vs 0.45 [0.41], P = .09, respectively). The taxonomic composition, alpha, and beta diversity remained consistent at all time points in both groups. The rate of patients reporting therapy-induced ocular symptoms and the mean discomfort grade were greater in group A than in group B (97% vs 26% and 4.97±2.48 vs 0.66±1.53, respectively). CONCLUSIONS: Compared with PVI 0.6%, CLX 0.02% induced a greater reduction of ocular surface bacterial load, with no significant alterations of the taxonomic composition. Moreover, CLX was better tolerated than PVI.

Antibiotic resistance of bacterial pathogens isolated from the conjunctiva in the Antibiotic Resistance Monitoring in Ocular micRoorganisms (ARMOR) surveillance study (2009-2021).

Antibiotic resistance in bacterial ocular infections is of significant clinical concern and may affect treatment outcomes. We report on in vitro antibiotic susceptibility rates and trends among conjunctival-sourced isolates collected in the Antibiotic Resistance Monitoring in Ocular micRoorganisms (ARMOR) surveillance study. A total of 2214 conjunctival isolates (918 Staphylococcus aureus, 589 coagulase-negative staphylococci [CoNS], 194 Streptococcus pneumoniae, 171 Pseudomonas aeruginosa, and 342 Haemophilus influenzae) obtained between 2009-2021 were analyzed. Staphylococci were commonly resistant to azithromycin (≥54.8%) and oxacillin (≥29.3%). Resistance among S. pneumoniae isolates was notable for azithromycin (34.0%) and penicillin (28.9%), while P. aeruginosa and H. influenzae isolates were highly susceptible to most tested antibiotics. Methicillin-resistant staphylococci demonstrated greater concurrent resistance to other antibiotics than methicillin-susceptible isolates and exhibited high rates of multidrug resistance (≥74.0%). Among staphylococci, antibiotic resistance increased with patient age, and there were small decreases in resistance to several drugs over the 13-year period. These findings indicate that resistance to antibiotics routinely used in ophthalmic practice remains high among conjunctival isolates.

Ocular microbial dysbiosis and its linkage with infectious keratitis patients in Northwest China: A cross-sectional study.

OBJECTIVES: To evaluate the alteration of ocular surface microbiome of patients with infectious keratitis in northwest of China. METHODS: The corneal scrapings, eyelid margin and conjunctiva samples were collected from 57 participants, who were divided into bacterial keratitis, fungal keratitis, viral keratitis and control group. The V3-V4 region of bacterial 16S rDNA in each sample was amplified and sequenced on the Illumina HiSeq 2500 sequencing platform, and the differences among different groups were compared bioinformatically. RESULTS: Significant alterations of the microbiome were observed in alpha-diversity and beta-diversity analysis between the keratitis groups and the control group (p < 0.05). There was no significant differences between eyelid margin and conjunctiva samples in Alpha-Diversity analysis, but a significant difference between eyelid margin and corneal scraping samples in the keratitis group (p < 0.05, independent t-test). The abundances of Bacillus, Megamonas, Acinetobacter, and Rhodococcu were significantly elevated, while the abundance of Staphylococcus was decreased in the keratitis group compared to the control group. CONCLUSIONS: The abundance of the ocular microbiome in patients with bacterial keratitis, fungal keratitis, or viral keratitis was significantly higher than those in the control group. Keratitis patients may have ecological disorder on ocular surface microbiome compared with controls. We believe that the conjunctiva and eyelid margin microbiome combined analysis can more comprehensively reflect the composition and abundance of ocular surface microbiome.

Conjunctival sac flora and drug susceptibility analysis in normal children in East China.

PURPOSE: To investigate the distribution characteristics of conjunctival sac flora and assess the susceptibility of commonly used topical antimicrobial agents in normal children under the age of 18 in East China. METHODS: In 2019, a study was conducted at Qingdao Eye Hospital of Shandong First Medical University to analyze the microorganism cultures of conjunctival sac in 1258 normal children (2516 eyes; average age, 6.21 ± 3.78 years) in East China. Exclusion criteria included children with ocular surface diseases and those who had used any topical antimicrobial agents recently. The microorganism species in the conjunctival sac were analyzed using the M-38A protocol (microdilution method; investigators read the minimum inhibitory concentration [MIC] values) by the Clinical and Laboratory Standards Institute to determine drug susceptibility. RESULTS: The incidence of conjunctival sac microorganism in children was 32.87% (827/2516), a total of 541 cases (male 293, female 248). Children with conjunctival sac flora in a single eye were 255 and in both eyes were 286 (no statistical difference, P > 0.05). The concordance rate of children with binocular conjunctival sac flora was 32.16% (174/541; male 84, female 90). A total of 42 species of bacteria were detected. Children with Gram-positive cocci accounted for the highest proportion, 91.54% (757/827). The top three bacteria with the highest detection rates were Staphylococcus epidermidis (S. epidermidis; 52.12%), Streptococcus (12.09%), and Staphylococcus aureus (S. aureus; 10.76%). Streptococcus mitis (5.20%) accounted for the highest proportion of Streptococcus.S. epidermidis had the highest proportion in all age groups and was positively correlated with age (r = 0.89, P = 0.03). Before six years of age, the streptococcal proportion(mainly S. mitis) was greater than that of Staphylococcus aureus. The drug susceptibility analysis showed that S. epidermidis was most sensitive to gatifloxacin (98.61%), while it had the highest resistance rate to erythrocin (87.94%). S. aureus had the highest susceptibility to moxifloxacin (100%). Streptococcus was most sensitive to moxifloxacin (96.97%) and had the highest resistance rate to tobramycin (92.93%). CONCLUSIONS: Conjunctival sac flora in children was dominated by Gram-positive cocci, mainly S. epidermidis, S. aureus, and Streptococcus. S. epidermidis increased with age; the proportion of Streptococcus was higher than S. aureus among children aged 0-6 years. The typical conjunctiva sac flora was generally sensitive to quinolones, such as moxifloxacin and gatifloxacin; Streptococcus displayed high resistance to tobramycin antibiotics; and the female children had higher resistance to tobramycin than the male children.

Conjunctival microbiome changes in soft contact lens users and contact lens discomfort patients.

INTRODUCTION: Contact lens discomfort (CLD) acts as a challenging problem, and the associated conjunctival microbiome changes were unclear. MATERIAL AND METHODS: Conjunctival sac swab samples were collected from 12 eyes of nonwearers (NW), 12 eyes of asymptomatic contact lens (ACL) wearers, and 11 eyes of CLD. The V3-V4 region of the 16S rRNA gene sequencing was used to investigate differences among three groups. RESULTS: No differences in alpha diversity were observed among the three groups. The beta diversity showed a distinct microbiome composition between ACL and CLD group (P = 0.018) with principal coordinate analysis. The relative abundance of Firmicutes was significantly higher in CLD (48.18%) than in ACL (13.21%) group (P = 0.018). The abundance of Bacillus in patients with ACL (0.05%) or with CLD (0.02%) were significantly lower than that in the NW (1.27%) group (P = 0.024, 0.028, respectively). Moreover, the abundance of Firmicutes was positively correlated with the OSDI scores in CLD patients (r = 0.817, P < 0. 01, Spearman). DISCUSSIONS: Patients with CLD have various degrees of bacterial microbiota imbalance in the conjunctival sac, compared with NW and ACL groups. CONCLUSION: Firmicutes may serve as a potential biomarker for the CLD patients.

In the current study, we investigated the conjunctival microbiome changes among nonwearers (NW), asymptomatic contact lens (ACL) wearers, and contact lens discomfort (CLD) patients using high-throughput 16S rRNA gene sequencing, and correlated relative abundances of the microbiota with clinical parameters.The relative abundance of Firmicutes was higher in CLD than that in ACL group. The abundance of Bacillus was lower in ACL or CLD group than that in NW group. The abundance of Firmicutes was positively correlated with the OSDI scores in CLD patients.Firmicutes may serve as a potential biomarker for the CLD patients.

The conjunctival fungal microflora of horses in a North Queensland tropical environment and their in vitro susceptibilities to antifungal agents.

Fungi are ubiquitous in the environment and part of the commensal microflora on the conjunctiva of equine eyes. North Queensland, being tropical, presents an ideal environment for fungi growth. When the cornea is injured, fungi can invade the corneal stroma, resulting in keratomycosis. The objectives of this study were to determine the fungal species specific to the eyes of horses in the Townsville region; to investigate the potential risk factors associated with the presence of fungi; and to test their susceptibility to antifungals to create an empirical guide for treatment. The eyes of forty ophthalmologically normal horses from James Cook University were sampled throughout the summer months of December 2017, January 2018, and January and February 2020. Cultured fungi were identified morphologically, and their identity confirmed by comparing partial 18sRNA DNA sequences with the NCBI nucleotide database. Minimum inhibitory concentration testing of common antifungal medications was performed. Sixty-one out of eighty conjunctival samples grew fungi, and 21 different fungi genera were isolated. The most common genera were Aspergillus (18%, 26/141), Curvularia (14%,20/141), Rhodotorula (12%,17/141) and Penicillium (12%,17/141). No significant association was found between age or environmental factors and fungal culture status. Most fungi were highly susceptible to voriconazole and ketoconazole but resistant to fluconazole and amphotericin B. This adds to the body of evidence on which species of fungi are present as normal ocular microflora of horses living in tropical regions of Australia, and an avenue for treating them.

Ocular conjunctival microbiome profiling in dry eye disease: A case control pilot study.

Purpose: Keratoconjunctivitis sicca (KCS) or dry eye disease (DED) is a multifactorial disease that results in discomfort, visual disturbance, and tear film instability with potential damage to the ocular surface. A pilot study was undertaken to determine if there were any major substantial differences in the ocular microbiome in DED patients versus healthy controls. Methods: The bacterial communities residing in the conjunctiva of patients with DED (n = 4) and healthy controls (n = 4) were assessed by 16S ribosomal RNA (rRNA) gene sequencing of the V4-V5 region. Results: The phyla Proteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes were most dominant and accounted for 97% and 94.5% of all bacterial sequences in patients and controls, respectively. At the genus level, 27 bacterial genera were found with more than two-fold difference between patients and controls. Four of these - Acinetobacter, Corynebacterium, Lactobacillus, and Pseudomonas spp. - dominated the ocular microbiome of all subjects, but were proportionately lower in DED (16.5%) compared to controls (37.7%). Several bacterial genera were found to be unique in DED (34) and controls (24). Conclusion: This pilot study is an attempt to profile the ocular microbiome in patients with DED that demonstrated a higher concentration of microbial DNA compared to controls, with Firmicutes phyla dominating the bacterial population in patients with DED.

Cytological and microbiological evaluation of conjunctiva in camels with and without conjunctivitis.

OBJECTIVE: To describe the cytological analysis of conjunctiva from normal camels and camels with bacterial conjunctivitis. ANIMALS STUDIED: This study was conducted on 7 normal camels and 15 camels affected with conjunctivitis. The affected camels had a history of conjunctivitis with signs including chemosis, blepharospasm, frequent blinking, and mild-to-moderate serous, mucoid, or purulent ocular discharge. PROCEDURES: Bacterial swabs were collected from the inferior conjunctival sac of the affected eye without topical anesthetics. Conjunctival smears were obtained from the conjunctival surface for cytological analysis. RESULTS: The cellular analysis of ocular smears revealed a higher percentage of basal cells, neutrophils, eosinophils, and macrophages in camels with conjunctivitis compared with normal camels. In contrast to this, smears from normal camels showed an increased percentage of superficial epithelial cells compared with affected camels. The microbiological assessment of conjunctival swabs collected from affected animals identified a bacterial growth of Staphylococcus aureus., Bacillus sp., Streptococcus sp., Enterococcus faecium., Staphylococcus sp., Corynebacterium sp., Coryne pseudotuberculosis., Saprophytica, Enterobacter cloacae, Escherichia coli, Proteus mirabilis, Proteus vulagaris, and Pseudomonas aeruginosa. CONCLUSIONS: It was observed that bacterial conjunctivitis in camels was associated with increased percentages of basal epithelial cells, neutrophils, eosinophils, and macrophages compared with normal camels, while normal camels showed an increased percentage of superficial epithelial cells compared with affected camels.

Effects of chloramphenicol, povidone-iodine 1% and 5% eye drops on the colonisation of conjunctival flora in patients undergoing cataract surgery.

Objectives: the aim was to compare 2 drops of either 5% chloramphenicol, 1% povidone-iodine or 5% povidone-iodine before cataract surgery on reducing the colonisation of bacterial flora in the conjunctiva. Design: This was a double-blind, randomised clinical trial study. Setting: Patients referred to Shahid Sadoughi Hospital in Yazd, Iran, for cataract surgery were studied. Participants: Totally 260 patients were enrolled. Intervention: The affected lower fornix was gently sampled with a sterile swab and cultured on appropriate microbiological media. Then one of the 3 solutions mentioned above was instilled into the conjunctival sac of the cases in groups 1, 2 and 3, respectively. After thirty minutes, new conjunctival swabs were taken and cultured. Main outcome measures: The type of bacteria isolated and their colony-forming unit per mL (CFU/mL) number were primary end-points. The statistical tests of Phi and Cramer's V and Wilcoxon and Kruskal-Wallis were applied to evaluate the relationship between the studied variables and culture results as the secondary end-point. Results: The studied patients were 129 (49.6%) males and 131 (50.4%) females. Bacterial growth was observed in 49 cases (18.85%); the most commonly isolated bacteria were Staphylococcus epidermidis (71.42%). In the povidone-iodine 5% and chloramphenicol groups (but not the povidone-iodine 1%), the decrease in the number of CFU/mL was statistically significant (P = 0.032 and P = 0.005, respectively, Wilcoxon test). Conclusion: A single dose of povidone-iodine 5% and chloramphenicol effectively reduces the colonisation of normal conjunctival bacteria and can be used as effective prophylaxis. Funding: This study was part of an MSc thesis of Nasrin Tofighi. Shahid Sadoughi University of Medical Sciences, Yazd, Iran, funded this work.

Prospective evaluation of the prevalence of conjunctival and intraocular bacteria in dogs undergoing phacoemulsification following a standardized aseptic preparation with 0.5% povidone iodine.

PURPOSE: To evaluate bacterial contamination of conjunctiva and aqueous humor in dogs undergoing phacoemulsification following asepsis with 0.5% povidone iodine and determine the influence of intravenous antibiotics on outcome of contamination. METHODS: Client-owned dogs were prospectively enrolled and randomly assigned to a control group, receiving 22 mg/kg intravenous cefazolin at induction prior to sampling, or experimental group receiving no antibiotic prior to sampling, masked to the surgeon. Dogs receiving antimicrobials in the pre-operative period were excluded. Asepsis was performed on all operated eyes using 0.5% iodine with minimum 3 min contact time at induction of anesthesia and repeated before surgery. A conjunctival swab and aqueous humor sample were collected prior to incision and following incision closure, respectively. Samples were submitted for aerobic and anaerobic bacterial culture and susceptibility. RESULTS: Seventy-one eyes of 42 dogs were included. Median age was 9 years. Thirty-nine and 32/71 eyes received intravenous cefazolin and no antibiotic, respectively. Median procedure time was 40 min per eye. Conjunctival cultures were positive in 6 eyes (8.5%): Serratia marcescens (5 eyes) and Cutibacterium acnes (1 eye). Aqueous humor cultures were positive in 5 eyes (7.0%): S. marcescens (2 eyes), Pseudomonas aeruginosa (2 eyes), Staphylococcus pseudointermedius (1 eye). Prevalence of positive culture did not differ between groups (p = .74), order of eyes for bilateral procedures (p = .74) and diabetic status (p = 1). CONCLUSIONS: Bacterial contamination of the conjunctiva and aqueous humor was present in 8.5% and 7.0% of dogs undergoing phacoemulsification after asepsis. Lack of IV cefazolin was not significantly associated with positive culture.

Conjunctival sac bacterial culture of patients using levofloxacin eye drops before cataract surgery: a real-world, retrospective study.

BACKGROUND: The use of antibiotics preoperatively is effective to decrease the incidence of ocular bacterial infections but may lead to high resistance rate, especially on patients with multi-risk clinical factors. This study systematically analyzed real-world data (RWD) of patients to reveal the association between clinical factors and conjunctival sac bacterial load and offer prophylaxis suggestions. METHODS: We retrieved RWD of patients using levofloxacin eye drops (5 mL: 24.4 mg, 4 times a day for 3 days) preoperatively. Retrieved data included information on the conjunctival sac bacterial culture, sex, presence of hypertension and diabetes mellitus (DM), and history of hospital-based surgeries. Data was analyzed using SPSS 24.0. RESULTS: RWD of 15,415 cases (patients) were retrieved. Among these patients, 5,866 (38.1%) were males and 9,549 (61.9%) females. 5,960 (38.7%) patients had a history of hypertension, and 3,493 (22.7%) patients had a history of DM. 7,555 (49.0%) patients had a history of hospital-based operations. There were 274 (1.8%) positive bacterial cultures. Male patients with hypertension and DM may be at increased risk of having positive bacterial cultures (P < 0.05). Staphylococcus epidermidis (n = 56, 20.4%), Kocuria rosea (n = 37, 13.5%), and Micrococcus luteus (n = 32, 11.7%) were the top 3 isolated strains. Most bacterial strains were resistant to various antibiotics except rifampin, and 82.5% (33 of 40 isolates) of Staphylococcus epidermidis isolates had multidrug antibiotic resistance. Numbers of culture-positive Staphylococcus epidermidis isolates in the male group and non-DM group were greater than those in the female and DM groups, respectively. Micrococcus luteus (n = 11, 8.8%) was found less frequently in non-hypertension group than in hypertension group. CONCLUSION: Sex (Male) and the presence of hypertension and DM are risk factors for greater conjunctival sac bacterial loads. We offer a prophylactic suggestion based on the combined use of levofloxacin and rifampin. However, this approach may aggravate risk of multidrug resistance.

Description of non-brachycephalic canine conjunctival microbiome before and after application of an antiseptic preparation.

OBJECTIVE: To evaluate canine conjunctival microbiome before and after an antiseptic preparation using aerobic culture and DNA sequencing. ANIMALS STUDIED: Six healthy non-brachycephalic dogs. PROCEDURES: Dogs randomly received povidone-iodine 1:50 dilution solution in one eye with the second eye serving as a control. Standardized volumes of solution, number of sterile cotton tipped applicators, and preparation times of 5 min were used. Aerobic culture and DNA sequencing samples were collected from inferior conjunctival fornices at baseline, immediately following, 24 h, and 4 weeks following antiseptic preparation. Aerobic culture data were evaluated using paired t-test and linear regression. Illumina platform was used to sequence bacterial DNA using primers to target the V4 region of bacterial 16S rRNA. Quantitative Insights Into Molecular Ecology (QIIME 2.0) was used to analyze data. RESULTS: Baseline aerobic cultures consisted of Bacillus (35%), Staphylococcus (30%), Streptococcus (20%), Moraxella (5%), Micrococcus (5%), and Simonsiella spp. (5%). No significant difference was detected in mean number of species cultured at baseline and following treatment (p = .465). Treatment, sample time, or interaction was not significant (p > .393). The most abundant phyla at baseline via DNA sequencing were Proteobacteria (57.04%), Actinobacteria (19.89%), Firmicutes (3.25%), and Bacteroidetes (5.5%). Alpha and beta diversity matrices at baseline and over time revealed no significant change in species richness or bacterial composition. Relative abundance of bacterial taxa did not significantly differ in treated or control eyes over time. CONCLUSIONS: Conjunctival bacterial flora identified differed between culture dependent and independent methods. The bacterial community remained stable over time after application of the antiseptic preparation.

Profiling of the Conjunctival Bacterial Microbiota Reveals the Feasibility of Utilizing a Microbiome-Based Machine Learning Model to Differentially Diagnose Microbial Keratitis and the Core Components of the Conjunctival Bacterial Interaction Network.

Both healthy and diseased human ocular surfaces possess their own microbiota. If allowed, opportunistic pathogens within the ocular microbiota may cause microbial keratitis (MK). However, the nonpathogenic component of the ocular microbiota has been proven to undermine the performance of culture, the gold standard of the etiological diagnosis for MK. As the conjunctival bacterial microbiota generates unique alterations with various oculopathies, this study aimed to evaluate the feasibility of distinguishing MK using machine learning based on the characteristics of the conjunctival bacterial microbiome associated with various types of MK. This study also aimed to reveal which bacterial genera constitute the core of the interaction network of the conjunctival bacterial microbiome. Conjunctival swabs collected from the diseased eyes of MK patients and the randomly chosen normal eyes of healthy volunteers were subjected for high-throughput 16S rDNA sequencing. The relative content of each bacterial genus and the composition of bacterial gene functions in every sample were used to establish identification models with the random forest algorithm. Tenfold cross validation was adopted. Accuracy was 96.25% using the bacterial microbiota structure and 93.75% using the bacterial gene functional composition. Therefore, machine learning with the conjunctival bacterial microbiome characteristics might be used for differentiation of MKs as a noninvasive supplementary approach. In addition, this study found that Actinobacteria, Lactobacillus, Clostridium, Helicobacter, and Sphingomonas constitute the core of the interaction network of the conjunctival bacterial microbiome.

Bacteriological profile of conjunctiva bacterial Flora in Northeast China: a hospital-based study.

PURPOSE: To investigate the distribution and influencing factors of preoperative conjunctival flora from patients undergoing penetrating ocular surgeries in northeast China. METHODS: An observational and cross-sectional study design was used. In 305 eyes of 305 patients without infective eye diseases who underwent ocular surgeries at the First Affiliated Hospital of Harbin Medical University between May 2018 and May 2019, conjunctival sac scrapings were collected on the day before surgery. RESULTS: The mean age of the participants was 60.73 ± 14.23 years, with the majority being female, married and unemployed with at least primary school education. The positive culture rate was 48.20% (147/305 eyes), and 191 bacterial strains were isolated; two or more strains were isolated from 22.45% (33/147) of positive samples. The most commonly isolated bacteria was Staphylococcus epidermidis (64.92%), surpassing Staphylococcus aureus (5.76%). The culture positive rate of the young (18-40 years) group was different between the females (26.67%) and males (69.23%) (P = 0.024), but in the middle-aged group and the elderly group, the rates between the sexes were similar, with an increasing trend. Patients who visited in summer or autumn presented a higher positive rate than other seasons. Hypertensive women had a higher rate than hypertensive men (58.14% vs. 40%, χ2 = 5.8662, P = 0.0154). CONCLUSIONS: In northeastern China, coagulase-negative Staphylococcus were the most common preoperative conjunctival bacteria. Hypertensive female patients, elderly patients, or those operated on in the summer and autumn should pay more attention to perioperative treatment. TRIAL REGISTRATION: ChiCTR2100044659 .

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.

Matrix metalloproteinase 9 is associated with conjunctival microbiota culture positivity in Korean patients with chronic Stevens-Johnson syndrome.

BACKGROUND: Stevens-Johnson syndrome (SJS) is an abnormal immune-response causing extensive exfoliation of the mucocutaneous tissue including conjunctiva. While several factors are associated with the alteration of conjunctival microbiota, the conjunctiva of SJS patients are found to harbor a different microbiota compared to healthy subjects. We investigated the conjunctival microbiota of Korean SJS patients, and identified factors associated with the conjunctival microbiota and its positive culture. METHODS: Medical records were retrospectively reviewed in 30 chronic SJS patients who had undergone conjunctival swab culture sampling. Demographic factors, chronic ocular surface complications score (COCS), tear break-up time (TBUT), tear secretion, tear matrix metalloproteinase 9 (MMP9), and results of conjunctival swab culture were assessed. RESULTS: Positive culture was seen in 58.1%. Gram positive bacteria was most commonly isolated, among which Coagulase-negative Staphylococci (45.5%) and Corynebacterium species (40.9%) were predominantly observed. Tear MMP9 positivity was observed significantly more in the positive culture group (100%) compared to the negative culture group (70%) (P = 0.041). Topical cyclosporine and corticosteroid were not associated with repetitive positive cultures. No significant differences in COCS, TBUT, and tear secretion were found between culture-positive and culture-negative groups. CONCLUSION: Our study suggests that tear MMP9 positivity may be related with the presence of an abnormal ocular surface microbiota in chronic SJS patients.

Bacterial distribution on the ocular surface of patients with primary Sjögren's syndrome.

Many studies have shown that gut microbial dysbiosis is a major factor in the etiology of autoimmune diseases but none have suggested that the ocular surface (OS) microbiome is associated with Sjögren's syndrome (SS). In this prospective study, we analyzed bacterial distribution on the OS in patients with primary SS. Among the 120 subjects included in this study, 48 patients (group A) had primary SS, whereas 72 subjects (group B) had dry eye symptoms that were unrelated to SS. We evaluated clinical dry eye parameters such as the OS disease index, ocular staining score (OSS), Schirmer's I test, and tear break-up time (TBUT). Conjunctival swabs were used to analyze the microbial communities from the two groups. Bacterial 16S rRNA genes were sequenced using the Illumina MiSeq platform, and the data were analyzed using the QIIME 1.9.1 program. The Shannon index was significantly lower in group A than in group B microbiota (p < 0.05). An analysis of similarity using the Bray-Curtis distance method found no difference in beta-diversity between the two groups (p > 0.05). In group A, Actinobacteria at the phylum level and Corynebacteria at the genus level exhibited low abundance than group B, but the differences were not statistically significant (p > 0.05). SS apparently decreases the diversity of the OS microbial community. These observations may be related to the pathophysiology of SS and should be investigated in future studies.

The eye of the giant anteater (Myrmecophaga tridactyla): Ophthalmic examination findings and diagnostic tests.

OBJECTIVE: To describe ophthalmic examination and diagnostic values for Schirmer tear test (STT), intraocular pressure (IOP), corneal horizontal diameter (CHD), palpebral fissure length (PFL), fundoscopy, and palpebral conjunctiva microbiota from healthy giant anteaters. ANIMALS STUDIED: Twelve giant anteaters (Myrmecophaga tridactyla), 11 adults and one juvenile, five males and seven females. PROCEDURES: The animals were submitted to general anesthesia and ophthalmic evaluation with portable slit-lamp biomicroscope, Finoff transilluminator, and fundoscopy, as well as STT, bacterial culture from palpebral conjunctiva, rebound tonometry IOP, and measurement of PFL and CHD. Data compiled were analyzed with ANOVA and Tukey tests. RESULTS: The results (mean ± standard deviation) were as follows: STT 8.04 ± 6.21 mm/min; IOP 10.92 ± 2.45 mmHg; PFL 0.75 ± 0.11 cm; CHD 0.96 ± 0.10 cm. Out of the 24 eyes swab samples for bacterial culture, 17 were positive, with three genera of Gram-positive bacteria identified Staphylococcus spp., Bacillus sp., and Corynebacterium sp. Gram-negative bacteria were not isolated from any of the samples. CONCLUSIONS: As conservation work in this vulnerable species continues, this report on basic ophthalmic examination and diagnostic parameters will be helpful improve their treatment and care. More ophthalmic studies are encouraged in animals within the Pilosa order.