Multiple use of preservative-free single dose unit dexamethasone 0.1% eye drops is safe within 24 hours.

BACKGROUND: Unpreserved single-dose unit (SDU) eye drops are commonly used to avoid benzalkonium chloride-related toxicity. Although intended for single use, many patients report off-label repeated use of SDUs over a prolonged period. We investigated whether repeated use of dexamethasone 0.1% SDUs in the same patient increases the bacterial contamination rate. METHODS: We prospectively enrolled patients scheduled for inpatient corneal and glaucoma surgery receiving dexamethasone 0.1% SDU four times per day from the same vial. To assess contamination rates, one drop from the vial was cultured immediately after opening the SDU (t0), 10 hours later after four drop applications (t10) and 24 hours after opening without further drop applications (t24). Conjunctival swabs were taken before and after drop application. Contamination rate was assessed with a standard clinical culturing protocol without introducing a positive control. RESULTS: 110 eyes of 109 patients were evaluated. Drops collected immediately after opening the SDU (t0) were contaminated in 9/110 cultures (8.1%). At t10, 13/110 cultures were contaminated (11.8%; p=0.267) and 11/110 at t24 (10.0%; t24 vs t0; p=1.00). In 5 of 21 cases of contaminated drops at t10 and/or t24, the same isolates were cultured from the initial conjunctival swab and the SDU. In three cases, the same bacterial species was found in consecutive samples. CONCLUSION: The contamination rate of the SDU did not increase after multiple use within 24 hours. Contamination from fingertip flora was more likely than from ocular surface flora. Reuse of dexamethasone 0.1% SDU in the same patient within 24 hours appears to be safe.

Probiotic LB101 alleviates dry eye in mice by suppressing matrix metalloproteinase-9 expression through the regulation of gut microbiota-involved NF-κB signaling.

Tear matrix metalloproteinase (MMP)-9 is an inflammatory signal in patients with dry eye (DE). In the present study, to understand the action mechanism of probiotic LB101 (Lactobacillus plantarum NK151 and Bifidobacterium bifidum NK175 [4:1] mix) against DE, we investigated its effect on tear amount and inflammatory marker expression levels in mice with unilateral exorbital lacrimal gland excision/atropine-benzalkonium chloride application (EB) or fecal microbiota transplantation from mice with EB (eFMT). Oral gavage of LB101 increased EB-suppressed tear amount and decreased EB-induced blinking number. Furthermore, LB101 decreased EB-induced TNF-α, IL-1ß, and MMP-9 expression, TNF-α+ and NF-κB+CD11c+ cell populations, and edema in the conjunctiva, while EB-suppressed IL-10 and occludin expression increased. LB101 also decreased EB-induced TNF-α and IL-1ß expression and NF-κB+CD11c+ cell population in the colon. eFMT also decreased tear amount and increased blinking number in the transplanted mice. eFMT increased TNF-α, IL-1ß, and MMP-9 expression and TNF-α+ and NF-κB+CD11c+ cell populations in the conjunctiva and TNF-α and IL-1ß expression and NF-κB+CD11c+ cell populations in the colon. Oral gavage of LB101 increased eFMT-suppressed tear amount and decreased eFMT-induced blinking number. Furthermore, LB101 decreased TNF-α, IL-1ß, and MMP-9 expression, TNF-α+ and NF-κB+CD11c+ cell populations, and edema in the conjunctiva and TNF-α and IL-1ß expression and NF-κB+CD11c+ cell population in the colon, while eFMT-suppressed IL-10 and occludin expression decreased. Furthermore, LB101 increased eFMT-suppressed Muribaculaceae, Prevotellaceae, and Lactobacillaceae populations in the gut microbiota, while eFMT-induced Bacteroidaceae population decreased. These findings suggest that DE may cause gut dysbiosis, which may be a risk factor for DE, and LB101 may alleviate DE with gut inflammation by suppressing the expression of MMP-9 and proinflammatory cytokines TNF-α and IL-1ß with the regulation of gut microbiota-involved NF-κB signaling.

In vivo assessment of antimicrobial activity and toxicity of repeated 1% povidone-iodine applications vs a single 5% povidone-iodine application.

PURPOSE: To demonstrate the noninferiority in antimicrobial effectiveness of thrice-repeated applications of 1% povidone-iodine (PI) compared with a single application of 5% PI on the ocular surface in vivo and to evaluate potential toxicity of PI to the corneal epithelium. SETTING: Department of Ophthalmology, Shaare Zedek Medical Center, Jerusalem, Israel. DESIGN: Prospective, randomized, double-blind study. METHODS: Patients randomly received either a single application of 5% PI or repeated irrigations of 1% PI on the ocular surface preoperatively. Bacterial colony counts were quantitatively analyzed by taking standardized conjunctival swabs before and after irrigation, and differences in bacterial colony counts were compared between the 2 groups. The presence of superficial punctate keratopathy was evaluated in all patients by the National Eye Institute grading system. RESULTS: The study comprised 102 eyes of 102 patients. Both 1% and 5% PI yielded a marked decrease in bacterial colonies compared with before application ( P < .001). The repeated application of 1% PI showed a significantly lower bacterial count compared with the single application of 5% PI ( P < .05) and resulted in significantly less damage to the corneal epithelium compared with a single 5% PI application ( P < .001). CONCLUSIONS: Preoperative repeated applications of 1% PI effectively eradicated ocular surface bacteria equal to that of 5% PI with a less deleterious effect on the corneal epithelium. Our findings suggest that 1% PI may be a promising alternative to 5% PI.

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.

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.

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.

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.

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.

Changes in Conjunctival Microbiota Associated With HIV Infection and Antiretroviral Therapy.

Purpose: HIV infection is associated with a variety of ocular surface diseases. Understanding the difference of the ocular microbiota between HIV-infected and healthy individuals as well as the influence of antiretroviral therapy will help to investigate the pathogenesis of these conditions. Methods: A cross-sectional study was conducted on subjects including HIV-negative individuals, untreated HIV-infected individuals, and HIV-infected individuals with antiretroviral therapy. Conjunctival microbiota was assessed by bacterial 16S rRNA sequencing of the samples obtained from the conjunctival swab. Results: The microbial richness in ocular surface was similar in HIV-negative, untreated HIV-positive, and highly active antiretroviral therapy (HAART) subjects. The bacterial compositions were similar in the two HIV infection groups but were significantly different from the HIV-negative group. HAART changed the beta diversity of bacterial community as determined by Shannon index. CD4+ T cell count had no significant influence on the diversity of ocular microbiota in HIV-infected individuals. Conclusions: The data revealed the compositional and structural difference in conjunctival microbial community in subjects with and without HIV infection, indicating that HIV infection or its treatment, may contribute to ocular surface dysbiosis.

Understanding the Interactions Between the Ocular Surface Microbiome and the Tear Proteome.

Purpose: The purpose of this study was to explore the interplay between the ocular surface microbiome and the tear proteome in humans in order to better understand the pathogenesis of ocular surface-associated diseases. Methods: Twenty eyes from 20 participants were included in the study. The ocular surface microbiome was sequenced by whole-metagenome shotgun sequencing using lid and conjunctival swabs. Furthermore, the tear proteome was identified using chromatography tandem mass spectrometry. After compositional and functional profiling of the metagenome and functional characterization of the proteome by gene ontology, association studies between the ocular microbiome and tear proteome were assessed. Results: Two hundred twenty-nine taxa were identified with Actinobacteria and Proteobacteria being the most abundant phyla with significantly more Propionibacterium acnes and Staphylococcus epidermidis in lid compared to conjunctival swabs. The lid metagenomes were enriched in genes of the glycolysis lll and adenosine nucleotides de novo and L-isoleucine biosynthesis. Correlations between the phylum Firmicutes and fatty acid metabolism, between the genus Agrobacterium as well as vitamin B1 synthesis and antimicrobial activity, and between biosynthesis of heme, L-arginine, as well as L-citrulline and human vision were detected. Conclusions: The ocular surface microbiome was found to be associated with the tear proteome with a role in human immune defense. This study has a potential impact on the development of treatment strategies for ocular surface-associated diseases.

Elevated levels of Merkel cell polyoma virus in the anophthalmic conjunctiva.

The human ocular surface hosts a paucibacterial resident microbiome and virome. The factors contributing to homeostasis of this mucosal community are presently unknown. To determine the impact of ocular enucleation and prosthesis placement on the ocular surface microbiome, we sampled conjunctival swabs from 20 anophthalmic and 20 fellow-eye intact conjunctiva. DNA was extracted and subjected to quantitative 16S rDNA PCR, biome representational karyotyping (BRiSK), and quantitative PCR (qPCR) confirmation of specific organisms. 16S ribosomal qPCR revealed equivalent bacterial loads between conditions. Biome representational in silico karyotyping (BRiSK) demonstrated comparable bacterial fauna between anophthalmic and intact conjunctiva. Both torque teno virus and Merkel cell polyoma virus (MCPyV) were detected frequently in healthy and anophthalmic conjunctiva. By qPCR, MCPyV was detected in 19/20 anophthalmic samples compared with 5/20 fellow eyes. MCPyV copy number averaged 891 copies/ng in anophthalmic conjunctiva compared with 193 copies/ng in fellow eyes (p < 0.001). These results suggest that enucleation and prosthesis placement affect the ocular surface flora, particularly for the resident virome. As MCPyV has been shown to be the etiologic cause of Merkel cell carcinoma, understanding the mechanisms by which the ocular surface regulates this virus may have clinical importance.

Identification of differentially expressed genes and signaling pathways in human conjunctiva and reproductive tract infected with Chlamydia trachomatis.

BACKGROUND: Currently, Chlamydia trachomatis-specific host defense mechanisms in humans remain poorly defined. To study the characteristics of host cells infected early with Chlamydia trachomatis, we used bioinformatics methods to analyze the RNA transcription profiles of the conjunctiva, fallopian tubes, and endometrium in humans infected with Chlamydia trachomatis. METHOD: The gene expression profiles of GSE20430, GSE20436, GSE26692, and GSE41075 were downloaded from the Gene Expression Synthesis (GEO) database. Then, we obtained the differentially expressed genes (DEGs) through the R 4.0.1 software. STRING was used to construct protein-protein interaction (PPI) networks; then, the Cytoscape 3.7.2 software was used to visualize the PPI and screen hub genes. GraphPad Prism 8.0 software was used to verify the expression of the hub gene. In addition, the gene-miRNA interaction was constructed on the NetworkAnalyst 3.0 platform using the miRTarBase v8.0 database. RESULTS: A total of 600 and 135 DEGs were screened out in the conjunctival infection group and the reproductive tract infection group, respectively. After constructing a PPI network and verifying the hub genes, CSF2, CD40, and CSF3 in the reproductive tract infection group proved to have considerable statistical significance. CONCLUSION: In our research, the key genes in the biological process of reproductive tract infection with Chlamydia trachomatis were clarified through bioinformatics analysis. These hub genes may be further used in clinical treatment and clinical diagnosis.

The effect of recurrent povidone-iodine usage on conjunctival flora in diabetic patients undergoing intravitreal injections.

PURPOSE: The purpose was to evaluate the change in the microbiological profile of diabetic patients undergoing intravitreal injections for diabetic macular edema. METHODS: Patients were included in this prospective study when referred for the first time for intravitreal injection to treat diabetic macular edema. For each patient, conjunctival cultures were taken from the lower fornix of each eye prior to the povidone-iodine application and the intravitreal injection. An additional culture was taken from the treated eye 20 min after the injection. The same culture protocol was used for the two following injections of these patients. A later conjunctival culture was also taken a month after the last injection. RESULTS: Twenty-one eyes of 21 patients were included. The mean duration of diabetes was 13.7 ± 7.9 years. Prior to the first intravitreal injection, 33% of cultures were positive. Prior to the third intravitreal injection, 26% of cultures were positive (p = 0.63), and 1 month after the last injection, 18% of cultures were positive (p = 0.495). The mean HbA1C was 8.1% ± 1.7%. HbA1C of patients with positive cultures was 8.0% ± 1.1% at the first intravitreal injection and 8.2% ± 1.0% at the third intravitreal injection. This was compared with HBA1C in eyes with negative cultures: 7.4% ± 1.2% (p = 0.45) and 7.1% ± 1.0% (p = 0.14), respectively. CONCLUSION: Repeated intravitreal injection for diabetic macular edema with application of povidone-iodine 5% in diabetic patients did not lead to a significant change in the percentage of positive conjunctival cultures. Patients with higher HbA1C had a slight, non-statistically significant trend for positive cultures.

Assessment of conjunctival flora in eyes with lacrimal passage obstruction before and after successful dacryoendoscopic recanalisation.

BACKGROUND: To investigate conjunctival bacterial flora in eyes with lacrimal passage obstruction before and after dacryoendoscopic recanalisation with lacrimal tube intubation. METHODS: One-hundred fifty eyes with lacrimal passage obstruction that were successfully treated by dacryoendoscopic recanalisation were enrolled. Conjunctival sampling was done for each eye before and 4 months after surgery. The lower fornix was rubbed by a sterile cotton swab, and the collected samples were cultured with several agar plates. Colonies were differentiated and enumerated by standard bacteriological laboratory techniques. RESULTS: Positive bacterial growth was detected in 42.0% of all the samples before surgery, and the positivity rate significantly decreased to 26.0% after surgery (p=0.0051). The number of strains detected also decreased from 20 before surgery to 9 after surgery, especially pathogenic microorganisms decreased. In addition, drug-resistant bacteria such as penicillin-resistant Streptococcus pneumoniae and methicillin-resistant Staphylococcus aureus were detected in nine eyes before surgery but were detected only in one eye after surgery. Coagulase-negative Staphylococci and Corynebacterium spp., which are common in normal conjunctival flora, accounted for 46.5% of all the isolates before surgery and 80.9% after surgery, showing a significant increase in the rate after surgery (p<0.0001). CONCLUSIONS: This study showed that physiological recanalisation of lacrimal passage after dacryoendoscopic surgery significantly decreased the culture positivity rate of conjunctival sac and the number of microorganism strains detected. It also decreased the number of potentially pathogenic and drug-resistant bacteria and increased the percentages of indigenous bacteria, causing the normalisation of conjunctival flora.

Fluoroquinolone resistance of Staphylococcus epidermidis isolated from healthy conjunctiva and analysis of their mutations in quinolone-resistance determining region.

BACKGROUND: Staphylococcus epidermidis is the most common pathogen in postoperative endophthalmitis and causes various infectious eye diseases. However, there is very little information on fluoroquinolone antibiotic resistance to S. epidermidis identified in conjunctival microbe and analysis of related genes. Here, the authors investigated the rate of resistance to fluoroquinolones of Staphylococcus epidermidis isolated from normal conjunctival microbes and mutations in the quinolone-resistance determining region (QRDR). METHODS: 377 eye samples from 187 patients who underwent intravitreal injection and cataract surgery were included. Specimens were taken from the bilateral lower conjunctival sacs using a cotton swab and cultured. The cultures were identified using MALDI-TOP MS and gyrA, gyrB, parC, and parE gene mutations of QRDR were confirmed by DNA extraction from resistant strains of S. epidermidis with a micro-dilution method using ciprofloxacin, levofloxacin, and moxifloxacin. RESULTS: The culture positive rate was 61.8% (231) for 374 eye samples. Of the 303 total strains cultured, S. epidermidis was the most common with 33.7% (102). Ten types of gene mutations were observed in the resistant S. epidermidis of 21 strains. One-point mutation was observed mainly in gyrA and parC, and a small number of mutations were observed in parE in the form of a double point mutations. When there were multiple point mutations in both gyrA and parC, the highest minimum inhibitory concentration was observed. CONCLUSIONS: The quinolone resistance rate of S. epidermidis increased in comparison with previous studies, and resistant S. epidermidis showed mostly QRDR mutations, which were mainly found in gyrA and parC, and showed strong resistance when mutated in both genes.

A new ophthalmic formulation containing antiseptics and dexpanthenol: In vitro antimicrobial activity and effects on corneal and conjunctival epithelial cells.

Antibiotic resistance is increasing even in ocular pathogens, therefore the interest towards antiseptics in Ophthalmology is growing. The aim of this study was to analyze the in vitro antimicrobial efficacy and the in vitro effects of an ophthalmic formulation containing hexamidine diisethionate 0.05%, polyhexamethylene biguanide (PHMB) 0.0001% disodium edetate (EDTA) 0.01%, dexpanthenol 5% and polyvinyl alcohol 1.25% (Keratosept, Bruschettini, Genova, Italy) on cultured human corneal and conjunctival cells. The in vitro antimicrobial activity was tested on Staphylococcus aureus, Methicillin-Resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, Streptococcus pneumoniae, Streptococcus pyogenes and Streptococcus mitis. For each microbial strain 10 µL of a 0.5 MacFarland standardized bacterial inoculum were incubated at 25 °C with 100 µL of ophthalmic solution for up to 6 h. After different periods of time, samples were inoculated on blood agar with 5% sheep blood. Moreover, a 0.5 MacFarland bacterial inoculum was seeded in triplicate on Mueller-Hinton Agar or on Mueller-Hinton Fastidious Agar; then a cellulose disc soaked with 50 µL of ophthalmic solution was applied on the surface of agar and plates were incubated for 18 h at 37 °C, in order to evaluate the inhibition of bacterial growth around the disc. Human corneal and conjunctival epithelial cells in vitro were incubated for 5, 10 and 15 min with Keratosept or its components. The cytotoxicity was assessed through the release of cytoplasmic enzyme lactate dehydrogenase (LDH) into the medium immediately after exposure to the drugs; the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to evaluate the metabolic cell activity. Our results show that Keratosept ophthalmic solution gave an average logarithmic (log) reduction of bacterial load of 2.14 ± 0.35 within 6 h of exposure (p-value < 0.05 versus control saline solution). On agar plates, all microbial strains, excluding P. Aeruginosa, showed an inhibition zone of growth around the Keratosept-soaked discs. Keratosept and its components after 5 and 10 min did not show any cytotoxic effect on cultured corneal and conjunctival cells, and only after 15 min a significant reduction of cell viability and an increase of cytotoxicity compared to control (vehicle) was seen; dexpanthenol 5% and polyvinyl alcohol accelerated the wounding of corneal cells in vitro. In conclusion, Keratosept showed good antimicrobial activity on the tested strains; the ophthalmic solution and its components were safe and non-toxic for the corneal and conjunctival epithelial cells for 5 and 10 min at the concentrations analyzed, and dexpanthenol 5% and polyvinyl alcohol promoted the wounding of corneal cells.

RNA-Seq analysis of differentially expressed genes of Staphylococcus epidermidis isolated from postoperative endophthalmitis and the healthy conjunctiva.

Staphylococcus epidermidis (S. epidermidis) is one of the primary pathogens in postoperative endophthalmitis, which is a devastating complication of cataract surgery and often results in irreversible visual loss and even blindness. Meanwhile, it is the most frequently isolated commensal bacterium in the healthy conjunctiva. In this study, we investigated the differentially expressed genes (DEGs) of S. epidermidis isolated from the patients with postoperative endophthalmitis and the healthy conjunctiva to predict their functions and pathways by Illumina high-throughput RNA sequencing. Using genome-wide transcriptional analysis, 281 genes (142 upregulated and 139 downregulated genes) were found to be differentially expressed (fold change ≥ 2, p ≤ 0.05) in the strains from endophthalmitis. Ten randomly selected DEGs were further validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR). GO enrichment analysis suggested that more DEGs were associated with the thioredoxin system and iron ion metabolism. KEGG pathway analysis revealed that more DEGs were associated with the pathways of the two-component system and pyruvate metabolism. Moreover, the gene SE1634 code for staphylococcal toxin was significantly upregulated in S. epidermidis strains of the endophthalmitis, which might be directly responsible for the pathogenesis of endophthalmitis. In conclusion, this research is helpful for further investigations on genes or pathways related with the pathogenesis and therapeutic targets of S. epidermidis endophthalmitis.