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Purpose: To evaluate the efficacy of liposomal amphotericin B (L-AMB) for the treatment of fungal keratitis. Methods: Patients with fungal keratitis confirmed by potassium hydroxide (KOH) smear and/or confocal microscopy were administered topical L-AMB and randomized into three groups treated with three different formulations. The medication was administered two hourly till clinical improvement was achieved, followed by six hourly till complete resolution. The outcome measures were time to clinical improvement, resolution of epithelial defect, stromal infiltrate, hypopyon, extent and density of corneal opacity, neovascularization, and best corrected visual acuity (BCVA) at 3 months. Results: Mean age of the patients was 46.6 ± 14.8 years, and trauma with vegetative matter was the most common predisposing factor. Aspergillus flavus (36%) was the most common fungus cultured, followed by Fusarium (23%). Mean time to clinical improvement, time to resolution of epithelial defect, mean time to resolution of infiltrate, and time to resolution of hypopyon were 3.45 ± 1.38, 25.35 ± 8.46, 37.97 ± 9.94, and 13.33 ± 4.90 days, respectively, and they were comparable among the three groups. There was a significant difference between treatment failure and success cases in terms of days of presentation (P < 0.01), size of the epithelial defect (P-value 0.04), and infiltrate size at presentation (P-value 0.04). At 3 months follow-up, no statistically significant difference was noted in BCVA and mean scar size among groups. Conclusion: L-AMB in a gel form is an effective antifungal agent that promotes the healing of fungal ulcers with notably least vascularization and better tolerance. Trial registration number: CTRI/2020/04/024550

Purpose: To analyze the pattern of bacterial pathogens causing infective keratitis and their resistance to the recommended antibiotics over six years. Methods: It was a retrospective study of 9,357 cases of bacterial keratitis from January 2015 to December 2020, at a tertiary care ophthalmic center. A total of 9,547 corneal specimens were obtained from the study subjects. Demographic details of the patients, pathogenic bacteria isolated, and their antimicrobial susceptibility were noted and analyzed. Results: Bacterial pathogens were identified in 23.52% of the specimens. The most common isolates were coagulase-negative Staphylococci (60.75%), followed by Pseudomonas aeruginosa (14.23%), Staphylococcus aureus (13.92%), gram negative bacilli of the family Enterobacterales (8.64%), Streptococcus spp. (1.72%), Acinetobacter spp. (0.13%), and other non-fermenting gram-negative bacilli (0.57%). In Staphylococci, 55–80% of isolates were resistant to erythromycin, and 40–70% to fluoroquinolones, while no resistance was observed against vancomycin. 40–60% of isolates of P. aeruginosa were resistant to cephalosporins, 40–55% to fluoroquinolones, and 30–60% to aminoglycosides. Also, 40–80% of isolates of Enterobacterales were resistant to cephalosporins, and 50–60% to fluoroquinolones. Most gram-negative isolates were susceptible to carbapenems and polymyxin B. Conclusion: To the best of our knowledge, our study is the largest compilation of microbiological profile of bacterial keratitis from North India. It highlights the current trend of the bacterial pathogens that cause infectious keratitis. Staphylococci and Pseudomonas were found to be the most common pathogens. Increased resistance was seen against some of the commonly prescribed empirical antibiotics. Such evidence is useful for restructuring the empirical prescription practices from time to time.

Purpose: Real-life comparison of three intravitreal drug regimens used in cases of endophthalmitis at a tertiary care center in India. Methods: In this prospective, comparative study, patients of bacterial endophthalmitis were grouped according to intravitreal antibiotic drug regimens into Group 1 (ceftazidime and vancomycin), Group 2 (piperacillin + tazobactam and vancomycin), and Group 3 (imipenem and vancomycin). Forty-eight hours after injection nonresponding/worsening patients underwent vitrectomy. Vitreous samples were subjected to microbiological and pharmacokinetic tests. Results: A total of 64 patients were included and divided into Group 1: 29, Group 2: 20, and Group 3: 15 cases. Also, 75% of patients were post-surgical endophthalmitis, whereas 25% were post-traumatic. Improvement in vision (V90-0) and vision at 3 months (V90) were comparable between the three groups. Visual recovery was poorer in post-traumatic cases. In post-surgical cases, visual recovery was poorer in those presenting beyond 72 h of onset of symptoms (P = 0.0002). Polymerase chain reaction (PCR) positivity (66%) was higher than BACTECTM (33%) and culture (14%). Antibiotic resistance was comparable amongst the three groups. Most patients (62/64) further underwent vitrectomy. Ceftazidime and vancomycin achieved vitreous concentrations more than the minimum inhibitory concentration (MIC) at 48 h after the first injection. Conclusion: The choice of antibiotics did not affect the rate of vitrectomy and final vision in a real-life scenario. Ceftazidime and vancomycin can still be used as first-line intravitreal antibiotics owing to their comparable microbial sensitivity profile and adequate ocular bioavailability.

Purpose: Therapeutic contact lenses (TCL) are known to help in epithelial healing and decreasing pain after various corneal surgeries. However, literature lacks any data describing their use following Descemet's stripping automated endothelial keratoplasty (DSAEK) where intraoperative epithelial debridement is commonly performed. Here we study the efficacy and safety of TCL in patients undergoing DSAEK. Methods: In this prospective, randomized, controlled clinical trial. 40 eyes of 40 patients of pseudophakic bullous keratopathy undergoing DSAEK were enrolled and randomized into two groups, control (no TCL) and test (TCL). Primary outcome was time taken for epithelial healing and secondary outcomes were postoperative pain score, graft attachment, best spectacle-corrected visual acuity, and endothelial cell loss at 3 months. Results: Average time taken for epithelial healing was 3.35 ± 0.49 days in the test group and 4.95 ± 1.05 days in the control group (P < 0.001). Average pain scores in first operative week were significantly lower in the test group as compared to control (P < 0.001). Graft detachment occurred in eight patients in control group and two in test group (P = 0.03). Both rebubbling rates and average endothelial cell loss at 3 months were higher in the control group with P = 0.07 and 0.06 respectively. No contact lens-related adverse effects were noted during the study period. Conclusion: Use of TCL in DSAEK leads to faster epithelial healing and lesser postoperative pain. In addition, it may also contribute to lower rebubbling rates and endothelial cell loss.

Purpose: To compare the physical and microbiological characteristics of McCarey-Kaufman (MK), Cornisol, and Optisol-GS media and evaluate the outcomes of keratoplasty performed using corneas stored in these three media. Methods: The study involved 60 donor corneas which were distributed in 3 groups: MK, Cornisol, and Optisol-GS. Corneas in these groups were further analyzed based on the type of keratoplasty performed (full thickness versus endothelial keratoplasty). At baseline, the endothelial cell density and death to preservation time of donor corneas were recorded. Following keratoplasty, patients were evaluated on day 1, at 1 month, 3 months, and 6 months follow-up. Outcomes were assessed in terms of corrected distance visual acuity (CDVA), endothelial cell density, percentage endothelial cell loss, and corneal thickness. The storage media were also assessed for their physical quality and their microbiological characteristics. Results: Physical characteristics of all three media were found to be within normal limits. Mean CDVA was comparable among the 3 groups at 6-month follow-up. The absolute endothelial cell count values were significantly lower for corneas stored in MK medium (1873.7 ± 261.1 cells/mm2) compared to the Cornisol (2085.0 ± 230.3 cells/mm2) and Optisol-GS media [(2180.3 ± 217.2 cells/mm2) (P =

Purpose: To evaluate the outcomes of water-soluble intrastromal natamycin (IS-NTM) as an adjunct therapy for recalcitrant fungal keratitis. Methods: This was a prospective interventional pilot study in the setting of a tertiary eye-care center. Twenty eyes of 20 consecutive patients with microbiologically proven recalcitrant fungal keratitis (ulcer size >2 mm, depth >50%, and not responding to topical NTM for 2 weeks) were recruited. The selected patients were injected with a novel composition of IS-NTM (10 ug/0.1 mL, soluble natamycin) prepared aseptically in the ocular pharmacology department. All the patients continued using topical NTM suspension 5% 4-hourly until the ulcer healed. Repeat injections were undertaken after 72 h depending on the clinical response and all the patients were followed till 6 months. Results: The mean age of the patients was 40.42 ± 10.09 years. The mean duration of the presentation was 20.8 ± 5.1 days. The most commonly isolated organisms were Aspergillus sp. (12/20, 60%) and Fusarium sp. (8/20, 40%). No patient had iatrogenic perforation or precipitate formation after IS-NTM injection. The overall cure rate with IS-NTM was 95% (19/20 patients). The number of patients who healed with the 1st, 2nd, and 3rd injection was 13, 5, and 1, respectively. One (5%) had no response to treatment and was subjected to penetrating keratoplasty. The average time taken for the resolution of the epithelial defect, stromal infiltrates, and hypopyon was 34 ± 5.2 days, 35.3 ± 6.4 days, and 15 ± 2.5 days. Healing with deep vascularization and cataract was noted in 6/19 eyes (31%) and 13/19 eyes (68.42%), respectively. Conclusion: Intrastromal injection of a novel formulation of NTM holds a promising role as adjunctive therapy to topical NTM in the management of recalcitrant filamentous fungal keratitis. The preliminary results are encouraging and further studies are required to validate the results.

Chlamydia trachomatis, an obligate intraocular bacteria causing trachoma, adult and neonatal inclusion conjunctivitis, was the leading cause of blindness in the last century worldwide. Improvement in socioeconomic and living conditions, availability of antibiotics, and introduction of National Trachoma Control Programmes reduced the prevalence in developed countries, but it persisted in resource-poor settings of Africa and Asia, including India. In 2016, as per the WHO report, trachoma is restricted to 42 countries, causing blindness/visual impairment in ~1.9 million people. India is one of the five countries with nearly half of total active trachoma patients. Introduction of Global Elimination of Trachoma 2020 program by the WHO, using SAFE strategy (surgery for trachomatous trichiasis; Antibiotics for C. trachomatis; Facial cleanliness; and environmental improvement) greatly reduced the prevalence, but trachoma still persists in India. Global increase in the reproductive tract infection by C. trachomatis urogenital serotypes (D-K) has led to concurrent increase in C. trachomatis eye infections. Therefore, kerato eye infections due to chlamydial infections continue to be seen in hospitals. Over the years, there have been advances in laboratory diagnostics, in understanding the pathogenesis, tissue tropism, C. trachomatis genomics, and treatment modalities. Due attention and research is still needed for the study of C. trachomatis eye infections.

Purpose: To study the susceptibilities of Aspergillus species against amphotericin B in infectious keratitis and to find out if drug resistance had any association with the molecular characteristics of the fungi. Materials and Methods: One hundred and sixty Aspergillus isolates from the corneal scrapings of patients with keratitis were tested for susceptibilities to amphotericin B by broth microdilution method. These included Aspergillus flavus (64 isolates), A. fumigatus (43) and A. niger (53). Fungal DNA was extracted by glass bead vertexing technique. Polymerase chain reaction (PCR) assay was standardized and used to amplify the 28S rRNA gene. Single-stranded conformational polymorphism (SSCP) of the PCR product was performed by the standard protocol. Results: Of the 160 isolates, 84 (52.5%) showed low minimum inhibitory concentration (MIC) values (≤ 1.56 μg/ml) and were designated as amphotercin B-sensitive. Similarly, 76 (47.5%) had high MICs (≥ 3.12 μg/ml) and were categorized as amphotericin B-resistant. MIC 50 and MIC 90 values ranged between 3.12-6.25 μg/ml and 3.12-12.5 μg/ml respectively. A. flavus and A. niger showed higher MIC 50 and MIC 90 values than A. fumigatus. The SSCP pattern exhibited three extra bands (150 bp, 200 bp and 250 bp each) in addition to the 260 bp amplicon. Strains (lanes 1 and 7) lacking the 150 bp band showed low MIC values (≤ 1.56 μg/ml). Conclusion: A. niger and A. flavus isolates had higher MICs compared to A. fumigatus, suggesting a high index of suspicion for amphotericin B resistance. PCR-SSCP was a good molecular tool to characterize Aspergillus phenotypes in fungal keratitis.

Background: Staphylococcus epidermidis , a commensal of the conjunctival sac has been incriminated as the commonest etiological agent of bacterial keratitis. However, the pathogenic potential of this commensal organism is not clearly known. Aim: To determine any phenotypic, molecular markers of S. epidermidis pathogenicity in bacterial keratitis. Materials and Methods: A total of 382 corneal ulcer isolates of S. epidermidis and 87 S. epidermidis isolates from healthy eyes (controls) were studied. Speciation, biotyping and antibiotic sensitivity testing were performed by conventional methods. Tube slime and adherence tests were carried out by recommended techniques. Plasmid analysis was conducted by a standard protocol. Statistical Analysis: Chi-square test was employed for calculations. Results: Out of 382 corneal ulcer isolates (Pathogens) 284 (74.3%) belonged to biotypes I and II. Slime was detected in 164 (42.9%) of 382 pathogens vs. 21 (24.1%) of 87 controls ( P < 0.001). Sixty-five (39.6%) of 164 slime positive isolates were multidrug-resistant as compared to only 49 (22.4%) of 218 slime negative isolates ( P < 0.001). A significantly higher number i.e., 73.1% (120/164) of slime-producers possessed a 21Kb plasmid in contrast to only 53.2% (116/218) of nonslime-producers ( P < 0.001). Presence of this plasmid had a statistical correlation of low significance with multidrug resistance ( P =0.04). One hundred and seventy-two (45.0%) of 382 pathogens and 24 (27.6%) of the 87 controls were adherent to artificial surfaces ( P =0.003) and the majority of the adherent organisms (99/172, 57.6%) were slime producers ( P < 0.001). Conclusions: Slime was associated with multidrug resistance in corneal ulcer isolates of S. epidermidis . The 21Kb plasmid could determine virulence as it was responsible for slime production and adherence.

PURPOSE: To study the microbial agents, chiefly Chlamydia trachomatis and other bacteria, in neonatal conjunctivitis. METHODS: Conjunctival specimens from 70 newborns with conjunctivitis were subjected to bacterial culture and sensitivity testing, monoclonal antibody based C. trachomatis antigen detection test and species-specific Chlamydia antibody detection in the sera of babies and their mothers, by micro-immunofluorescence assay. RESULTS: Bacteria were isolated from 35 (50%) babies; the majority (20, 57.14%) were Staphylococcus epidermidis. C. trachomatis antigen was detected in conjunctival smears of 17 (24%) babies, and 6 (35.29%) of them were positive for other bacteria. Six babies and their mothers tested positive for C. trachomatis Ig G antibodies. At follow-up after 14 weeks, 6 (35.29%) of the Chlamydia antigen-positive babies were found to have developed recurrent conjunctivitis. CONCLUSION: C. trachomatis is responsible for almost a quarter of all cases of neonatal conjunctivitis, with recurrences in 35% of cases. Bacteria could be isolated from 50% of the patients though the exact role of Staphylococcus epidermidis, isolated from 28.65% of the neonatal conjunctivitis cases, remains unclear.

Purpose: To evaluate polymerase chain reaction (PCR) in diagnosis of Chlamydia trachomatis. Methods: In this study PCR assay was used to amplify the 517bp region of common endogenous plasmid in conjunctival specimens from 178 patients with follicular conjuntivitis, and to amplify the 1000bp region of major outer membrane protein (MOMP) gene of C.trachomatis from 71 of these 178 patients. The PCR-amplified products were visualised by agarose gel electrophoresis and ethidium bromide staining and Southern hybridisation with radio-labelled internal probes. The test was compared with a direct immunofluorescence assay using monoclonal antibody for Chlamydia antigen detection. Results: The plasmid PCR assay was positive in 95 (53.37%) of the 178 specimens processed whereas the Chlamydia antigen was detected in 69 (38.76%) of the 178 specimens by direct immunofluorescence assay (p= 0.005). In the 71 specimens processed for both the PCR assays, plasmid PCR was positive in 52 (73.23%) and MOMP PCR was positive in 43 (60.56%) of the specimens (p=0.10). Thirty seven of these 71 specimens which were positive in both PCR assays were also positive in direct immunofluorescence assay. Conclusion: The PCR assays could detect Chlamydia in a significantly larger number of specimens than conventional antigen detection assay, and being marginally more sensitive, the plasmid PCR assay has the potential for wider use in the diagnosis of trachoma.

Incidence of spinal neurocysticercosis (NCC) is rare. Isolated spinal NCC is still rarer. We present here a case report where a young lady presented with all the clinical features of pseudotumor cerebri (PTC), where medical treatment for PTC failed and the presence of cysticercous in spinal canal was detected only on the operation table, while doing a lumbo-peritoneal shunt (LP shunt) to save her vision. Diagnosis could be confirmed only after the histopathology report was received. She did not have any direct evidence of spinal involvement, thereby eluding correct diagnosis. In English literature, we could not find any report of isolated and silent spinal NCC associated with PTC. In addition, we could not find any report of recovery of cysticercous larva through the Touhey′s needle injury, although this was an incidental finding. In endemic areas, isolated spinal NCC should be suspected in patients presenting with PTC.

An epidemic of acute haemorrhagic conjunctivitis affecting persons of all ages and both sexes occurred in Delhi and surrounding areas during the monsoon season of 1994. The symptoms lasted on an average for 4-5 days. In some of the patients corneal involvement was observed. Conjunctival swabs from the affected patients were processed for viral antigen detection, virus isolation and bacterial culture and sensitivity. Viral antigen was detected in 62% (31/50) of the smears tested by indirect immunofluorescence assay. In 22 (44%) of the specimens Coxackie A 24 (Cox A 24) virus antigen and in 9 (18%) of the specimens Entero Virus 70 (EV 70) antigen were detected. In confluent monolayers of Hep 2 cells cytopathic virus was isolated in 10 (30.30%) of the 33 specimens processed. The isolated viruses were identified as either Cox A 24 (7 isolates) or EV 70 (3 isolates) using indirect immunofluorescence assay. Super added bacterial infection was observed in 33% (89/270) of the cases, Staphylococcus albus being the predominant bacteria isolated.

We describe a case with non-responding polymicrobial spontaneous corneal ulceration in an HIV-positive patient. Acanthamoeba was among the microorganisms isolated.

Purpose: Therapeutic contact lenses (TCL) are known to help in epithelial healing and decreasing pain after various corneal surgeries. However, literature lacks any data describing their use following Descemet's stripping automated endothelial keratoplasty (DSAEK) where intraoperative epithelial debridement is commonly performed. Here we study the efficacy and safety of TCL in patients undergoing DSAEK.Methods: In this prospective, randomized, controlled clinical trial. 40 eyes of 40 patients of pseudophakic bullous keratopathy undergoing DSAEK were enrolled and randomized into two groups, control (no TCL) and test (TCL). Primary outcome was time taken for epithelial healing and secondary outcomes were postoperative pain score, graft attachment, best spectacle-corrected visual acuity, and endothelial cell loss at 3 months.Results: Average time taken for epithelial healing was 3.35 ± 0.49 days in the test group and 4.95 ± 1.05 days in the control group (P < 0.001). Average pain scores in first operative week were significantly lower in the test group as compared to control (P < 0.001). Graft detachment occurred in eight patients in control group and two in test group (P = 0.03). Both rebubbling rates and average endothelial cell loss at 3 months were higher in the control group with P = 0.07 and 0.06 respectively. No contact lens-related adverse effects were noted during the study period.Conclusion: Use of TCL in DSAEK leads to faster epithelial healing and lesser postoperative pain. In addition, it may also contribute to lower rebubbling rates and endothelial cell loss. [ABSTRACT FROM AUTHOR]