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Purpose: To study the effect of prophylactic intracameral moxifloxacin (ICM) on microbiological profile and antimicrobial sensitivity of culture-proven postoperative endophthalmitis (POE)., Methods: This study evaluated culture-proven POE diagnosed within 6 weeks of cataract surgery over two periods, period-1 (January 2010-March 2015) and period-2 (April 2015-December 2019), before and after introduction of prophylactic ICM, respectively., Results: In period-1, 100 cases of culture-positive POE were reported (1 in 4879, 0.02%), while 20 cases (1 in 24635, 0.004%) were reported in period-2 (5-fold reduction, p < .001). The cumulative culture positivity rate decreased from 27.6% to 17.1% (1.6-fold) . Coagulase-negative Staphylococci (CoNS) were significantly reduced ( p  = .005)., Conclusions: ICM reduced the incidence of culture-proven POE, CoNS in particular. In future, POE caused by rarer pathogens may become more prevalent due to reduction in the rate of commoner and more virulent pathogens with use of intracameral antibiotics. The moxifloxacin sensitivity of CONS did not show change with the use of ICM. Studies with larger population of POE will be more helpful to understand the trend better.

Purpose: To evaluate the utility of galactomannan testing of intraocular fluid in the diagnosis of Aspergillus endophthalmitis (AE)., Methods: This retrospective study enrolled three groups of patients, including those with 17 eyes with AE; 20 eyes with intraocular infection of bacteria, viruses, or other fungi; and 19 eyes with cataract. Intraocular fluid from all these patients was collected for galactomannan testing. In addition, the receiver operating characteristic curves and diagnostic significance were analyzed., Results: The mean optical density index (ODI) of galactomannan was 5.77 ± 1.73 in the AE group, which was significantly higher than that in the non- Aspergillus intraocular infection group (0.19 ± 0.11, P < 0.001) and the negative control group (0.29 ± 0.27, P < 0.001). The area under the receiver operating characteristic curve (area under the curve) was 1.00 (95% confidence interval, 1.00-1.00; P < 0.001) in the AE group and the other two groups. At a cutoff optical density index of 1.88, the sensitivity and specificity were 100.0% and 100.0%, respectively, and the Youden index reached its highest value of 1.00., Conclusion: Galactomannan testing of intraocular fluid indicated good sensitivity and specificity for the diagnosis of AE, thereby promising a rapid diagnostic modality for AE.

Purpose: To explore the clinical value of β-D-glucan (BDG) testing and metagenomic next-generation sequencing (mNGS) for detecting the pathogens of fungal endophthalmitis (FE)., Methods: This study included 32 cases (32 eyes) with FE and 20 cases (20 eyes) with intraocular inflammation caused by other etiologies. All patients underwent extraction of aqueous humor or vitreous fluid samples for BDG testing and mNGS. The diagnostic performance and total clinical concordance rate of BDG testing and mNGS for FE were evaluated and calculated based on the results of the clinical diagnosis., Results: Among the clinically diagnosed FE, the positivity rates of BDG testing and mNGS (90.63%) were both significantly higher ( P < 0.001) than that of microbial cultures (53.13%). There was 100% consistency in pathogen identification using mNGS and culture identification for culture-positive cases. The area under the curve was 0.927 for BDG testing and 0.853 for mNGS. When the two tests were combined, sensitivity (93.75%), specificity (100.00%), and total clinical concordance rate (96.15%) were all improved, compared with the single tests., Conclusion: The positive rates of BDG test and mNGS were markedly higher than those of cultures in FE identification. The combination of these two tests showed improved performance when compared with individual tests.

The microbiome of the ocular surface has been characterised, but only limited information is available on a possible silent intraocular microbial colonisation in normal eyes. Therefore, we performed next-generation sequencing (NGS) of 16S rDNA genes in the aqueous humour. The aqueous humour was sampled from three patients during cataract surgery. Air swabs, conjunctival swabs from patients as well as from healthy donors served as controls. Following DNA extraction, the V3 and V4 hypervariable regions of the 16S rDNA gene were amplified and sequenced followed by denoising. The resulting Amplicon Sequence Variants were matched to a subset of the Ribosomal Database Project 16S database. The deduced bacterial community was then statistically analysed. The DNA content in all samples was low (0-1.49 ng/µL) but sufficient for analysis. The main phyla in the samples were Acinetobacteria (48%), Proteobacteria (26%), Firmicutes (14%), Acidobacteria (8%), and Bacteroidetes (2%). Patients' conjunctival control samples and anterior chamber fluid showed similar patterns of bacterial species containing many waterborne species. Non-disinfected samples showed a different bacterial spectrum than the air swab samples. The data confirm the existence of an ocular surface microbiome. Meanwhile, a distinct intraocular microbiome was not discernible from the background, suggesting the absence of an intraocular microbiome in normal eyes.

Purpose: This paper aimed to assess the diagnostic utility of a newly developed gene-based technology-nanopore targeted sequencing (NTS) in suspected endophthalmitis patients., Methods: This retrospective study included 43 patients (44 eyes) with suspected endophthalmitis. NTS was applied along with microbiological culture to detect unknown pathogens in intraocular fluid samples. The diagnostic utility of NTS was mainly evaluated from three aspects, including the positivity rate of bacterial/fungal presence, diagnostic turnaround time and the frequency of change in treatment based on etiology test results. Non-parametric, two-sided Wilcoxon rank sum test, the McNemar's test and the kappa statistic were used for statistical comparisons., Results: NTS showed significant advantages over traditional culture in positivity rates and diagnostic time (P < 0.001, kappa = 0.082; Z = -5.805, P < 0. 001). As regards antibiotic strategy, 17 patients (39.53%) and 5 patients (11.63%) underwent medication change following NTS and culture results respectively (P < 0.001, kappa = 0.335). With reasonable use of antibiotic and surgical intervention, most patients responded favorably, judged by significantly improved visual acuity (Z = -4.249, P < 0.001). The mean duration of hospitalization was 8.49 ± 2.45 days (range, 1-16 days)., Conclusion: The high efficiency feature of NTS in pathogen detection renders it a valuable supplementary to traditional culture. Additionally, it has facilitated patients' management for the early and precise diagnosis of endophthalmitis., (© 2023. The Author(s).)

Background: Leptospira species are difficult to culture. Thus, when there is suspicion for an infectious etiology to uveitis, bacterial cultures may fail to identify Leptospira. We describe a case of leptospirosis-associated uveitis that evaded culture and molecular assays. DNA sequencing of the aqueous fluid showed the presence of Leptospira spp., Methods: Retrospective case review of clinical and laboratory features of a patient with ocular leptospirosis is presented., Results: DNA sequencing identified the genome of Leptospirosis spp. in the aqueous humor., Conclusion: Metagenomic sequencing, by virtue of its unbiased nature, can be a helpful adjunctive test when a strong clinical suspicion for intraocular infection persists despite negative routine culture and molecular assays.

In Brief: Nanopore targeted sequencing showed a higher positivity rate and a shorter turnaround time than did traditional culture in identifying pathogens in the intraocular fluid samples of patients with endogenous endophthalmitis., Purpose: To evaluate the feasibility of clinical application of nanopore targeted sequencing (NTS) for the identification of pathogens in patients with endogenous endophthalmitis, especially those with fungus-associated endophthalmitis., Methods: In this retrospective study, medical records and etiological results of 27 patients (34 eyes) with endogenous endophthalmitis were reviewed. The intraocular fluid samples were examined using both NTS and microbial culture. The results included the differences in detection time, positivity rate of pathogen detection, and positivity rate of fungus identification between two methods., Results: NTS and microbial culture enabled the detection of etiologic agents in 89.28% and 35.71% of the samples, respectively. The difference of positivity rate between these methods was statistically significant ( P < 0.001). NTS also showed high sensitivity in both culture-positive and culture-negative samples (100% and 83.33%, respectively). Regarding culture-positive samples, the NTS results displayed a strong match with culture results. NTS showed a significantly higher positivity rate for fungal infection than did microbial culture (46.43% vs. 7.14%, P = 0.002). The average detection time of NTS was 1.11 ± 0.31 days, which was shorter than that of microbial culture (2.50 ± 0.58 days, Z = -4.686, P < 0.001). NTS technology facilitated an informed switch of intravitreal antimicrobial agents in 13 eyes., Conclusion: NTS, as a sensitive, specific, and timely complementary method, can be used along with traditional methods for the identification of pathogenic microorganisms in the intraocular fluid of patients with endogenous endophthalmitis., (Copyright © 2023 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the Opthalmic Communications Society, Inc.)

Purpose: To determine the bacterial spectrum of exogenous endophthalmitis of different origins, namely, posttraumatic, postcataract surgery, filtering bleb-associated, and intravitreal treatment-related endophthalmitis, using the 16S rDNA sequencing method., Methods: Aqueous humor or vitreous humor samples were collected from 24 endophthalmitis patients. Traditional cultivation and 16S rDNA sequencing were conducted with these samples. Three senile cataract controls and one intraocular irrigating solution were used as sequencing control., Results: Eleven of the 24 samples (45.8%) obtained positive bacterial cultivation, and each sample positive for only one species. The 11 culture-positive species could all be identified in their corresponding sequencing results, but only four strains being the top one pathogen in the sequencing. A total of 567 species were isolated using 16S rDNA sequencing, with the top five species being Pseudomonas sp., Staphylococcus epidermidis, Staphylococcus sp., Streptococcus sp., and Enterococcus faecalis. The dominant bacterial strains varied among the different endophthalmitis categories but with no significant difference in the overall bacterial spectrum. Bacterial atlas containing Pseudomonas, Streptococcus, Staphylococcus, Actinomycetales&#95;unclassified, Thermus, and Janibacter was shared by the four categories. Aqueous humor bacterial profile showed a higher overlap with contaminating bacteria from the environment., Conclusions: 16S rDNA sequencing is more efficient for endophthalmitis pathogen screening than the traditional cultivation method in terms of positive detection rate and the number of bacteria identified. But the risk of environmental contamination exists when using 16S rDNA sequencing method for endophthalmitis diagnosis. Different categories of endophthalmitis displayed diversified bacterial composition., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Purpose : The purpose of this report is to present an extremely rare case of endogenous Listeria monocytogenes endophthalmitis. Methods : A 72-year-old woman was diagnosed with endogenous endophthalmitis. Listeria monocytogenes was identified by culture of aqueous humor. Multiple investigations could not reveal any source of infection. An overview of other published cases is presented and a standardized treatment protocol is suggested. Results : Treatment consisted of topical therapy with antibiotics, corticosteroids, cycloplegics, and intraocular pressure-lowering agents, four intravitreal injections of vancomycin, an anterior chamber washout, intravenous amoxicillin, and eventually a pars plana vitrectomy. The inflammation subsided and the patient was discharged with a best-corrected visual acuity of 20/40. Conclusion : Our study emphasizes the importance of adequate microbiological testing and initiating appropriate treatment early in the management of endogenous endophthalmitis.

Purpose : The aim of this study was to evaluate and compare incidence and outcomes of management of acute endophthalmitis after microincision vitrectomy surgery (MIVS) and intravitreal injections (IVIs). Methods : Medical records were retrospectively reviewed from January 2012 to December 2017, and the incidence, clinical and microbiological profiles of acute endophthalmitis were documented. Results : Of 26,332 MIVS and 24,143 IVI performed, incidence of acute endophthalmitis in MIVS group was 0.027% (1 in 3761 cases) against 0.054% (1 in 1857 cases) in IVI. Gram-positive organisms were causative in post IVI group as against gram-negative organisms in MIVS group. Conclusion : Incidence of endophthalmitis after IVI is almost twice that after MIVS. A trend toward poorer outcomes in MIVS eyes was observed. Both MIVS and IVI being pars plana procedures warrant similar kind of aseptic precautions.

Purpose: To report the diagnosis and management of a series of chronic postoperative endophthalmitis caused by Stephanoascus ciferrii., Methods: This is a retrospective consecutive case series at a tertiary eye care institute. Case records of consecutive cases of chronic postoperative endophthalmitis caused by a novel fungus S. ciferrii were analyzed. The clinical presentations, microbiologic workup including microscopy, culture, antifungal susceptibility and polymerase chain reaction for fungal DNA of clinical samples, and the clinical management were noted. The cases underwent anterior chamber tap, vitreous biopsy, core vitrectomy, and intraocular lens explantation, and the management consisted of a combination of topical and systemic therapy and intravitreal injections of antimicrobial agents., Results: The report includes four eyes of four patients. All patients were diagnosed as chronic postcataract surgery endophthalmitis, and they showed variable levels of persistent low-grade intraocular inflammation till the intraocular lens was explanted. S. ciferrii (identified by the Vitek 2 compact system) was grown in culture from one or more of the clinical samples such as vitreous, anterior chamber fluid, capular bag, and intraocular lens. Fungal DNA was detected from vitreous in one case. One isolate tested for antifungal susceptibility was resistant to amphotericin B; however, the patient responded to treatment with the drug. Good clinical outcome was achieved in all patients., Conclusion: We describe successful treatment of endophthalmitis caused by S. ciferrii. A good response is obtained after pars plana vitrectomy and intraocular lens explantation.

Background : Endogenous fungal endophthalmitis is a sight-threatening condition with potentially devastating outcome. Hematogenous spread of the infective seedings is the route of infection. Infected individuals have usually a compromised immune status. The clinical picture of mycotic endogenous endophthalmitis is commonly seen as chorioretinitis. Candida is the most common fungus. Cladosporium causing endogenous endophthalmitis is a rare occurrence, with only a few cases published. Methods : The report includes study and management of a diabetic patient with endogenous cladosporium endophthalmitis mimicking toxoplasma retinochoroiditis. Results : Diagnosis was confirmed as Cladosporium Cladosporioides in vitreous and aqueous aspirate by polymerase chain reaction-based DNA sequencing. Patient was successfully managed with intravitreal and systemic voriconazole. Conclusion : Cladosporium can cause endogenous endophthalmitis and mimic toxoplasma retinochoroiditis. Vitreous biopsy can help in diagnosis in the absence of positive blood culture. Intravitreal voriconazole along with systemic voriconazole shows a good response.

Purpose : To evaluate immunoblot (IB) and polymerase chain reaction (PCR) to diagnose ocular- and neurosyphilis. Methods : Prospective cross-sectional study. Aqueous humor (AH) and cerebrospinal fluid (CSF) samples were tested for treponemal DNA or antibodies to treponemal antigens. Results : Thirteen of 106 cases had positive syphilis serology of which 69.2% were HIV+ (median CD4+ = 181 cells/µL). Four cases met CDC criteria for neurosyphilis (3 confirmed, 1 probable) and 2 additional cases required neurosyphilis treatment according to UpToDate algorithms. All AH and CSF samples tested PCR negative. Five cases were CSF IB+ and 3 cases AH IB+. Using our classification, eight patients had confirmed neurosyphilis, one had probable neurosyphilis, three had confirmed ocular syphilis and nine had probable ocular syphilis. Conclusion : Our findings suggest that IB of AH and CSF provides additional evidence to diagnose ocular and neurosyphilis and allows us to classify them as probable or confirmed.

Background: This study aimed to assess and compare the clinical value of aqueous humor polymerase chain reaction (PCR) and serologic tests in patients diagnosed with suspected infectious uveitis., Methods: In this retrospective observational study, data of 358 patients who were diagnosed with suspected infectious uveitis and who underwent aqueous humor PCR testing were analyzed. PCR and serologic test results were compared with the clinical features., Results: The rates of initial diagnoses for infectious uveitis were higher with PCR (99 patients, 28%) compared to those with serologic tests (38 pateints, 11%). The diagnostic positivity of PCR was 29% for anterior uveitis, 0% for intermediate uveitis, 5% for posterior uveitis, and 30% for panuveitis. In particular, PCR was useful in confirming the diagnosis of cytomegalovirus and varicella-zoster virus infections and Toxoplasma gondii-associated uveitis. For PCR test, the sensitivity was 0.431, specificity was 0.985, and the negative and positive predictive values were 0.506 and 0.980, respectively. For IgM test, the sensitivity was 0.151, specificity was 0.970, and the negative and positive predictive values were 0.403 and 0.895, respectively., Conclusion: Aqueous humor PCR can be a valuable diagnostic tool for confirming the infectious etiology in patients clinically diagnosed with uveitis. PCR had good predictive and diagnostic value for anterior uveitis and panuveitis compared with that for intermediate and posterior uveitis.

Purpose: To analyze practice patterns, microbiologic yield, and clinical outcomes in cases of presumed infectious endophthalmitis at a single tertiary academic referral center., Design: Retrospective chart review., Participants: Eyes diagnosed with endophthalmitis at the Duke Eye Center between January 1, 2009, and January 1, 2018, that underwent aqueous tap, needle vitreous tap, or mechanical vitreous biopsy with pars plana vitrectomy (PPV) in conjunction with intravitreal antimicrobials., Methods: Charts of 133 eyes of 130 consecutive patients were identified and reviewed for etiology of endophthalmitis, ocular history, interventions, visual outcomes, complication rates, and microbiologic culture results., Main Outcome Measures: Microbiologic yield and corrected visual acuity (VA) at initial presentation and last follow-up (up to 1 year) were the primary outcome measures. Secondary outcome measures included need for subsequent intervention, retinal detachment (RD) rate, and antimicrobial sensitivities., Results: Of the 133 cases of presumed infectious endophthalmitis, 74% were unrelated to cataract surgery. Initial management included intravitreal antimicrobials with aqueous tap in 46% of eyes, needle vitreous tap in 59% of eyes, and mechanical vitreous biopsy with PPV in 14% of eyes. Twenty-seven percent of cases that underwent initial aqueous or needle vitreous tap required subsequent intervention with PPV. Overall, 45% of endophthalmitis cases were culture proven. Needle vitreous tap and mechanical vitreous biopsy with PPV were more likely to yield culture growth compared with aqueous tap (P = 0.042, P = 0.004, respectively). All isolated bacteria were sensitive to the combination of intravitreal vancomycin and ceftazidime. Eyes that underwent initial mechanical vitreous biopsy with PPV had worse VA at presentation (P = 0.024) and at last follow up (P = 0.011) compared with eyes that underwent initial aqueous tap or needle vitreous tap. An RD occurred within 3-12 months in 13% (17/133) of cases, and of these, 41% had an endogenous etiology., Conclusions: Needle vitreous tap and mechanical vitreous biopsy were more effective in yielding positive cultures compared with aqueous tap; however, culture results may not influence initial intravitreal antimicrobial choice. Eyes with presenting VA between counting fingers and hand motion at 1 foot were more likely to undergo initial PPV., (Copyright © 2020 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Background: Endophthalmitis is a rare but potentially devastating complication of intravitreal injection. The causative organism plays an important role in prognosis following endophthalmitis. Here we present the first reported case of Turicella otitidis endophthalmitis, which is notable for a delayed presentation., Case Presentation: A 71 year old male who was receiving intravitreal aflibercept injections for neovascular age-related macular degeneration presented 4 weeks after his most recent intravitreal injection and was found to have endophthalmitis. Polymerase chain reaction (PCR) testing of aqueous fluid was positive for Turicella otitidis. The endophthalmitis responded well to treatment with intravitreal antibiotics., Conclusions: Coryneform bacteria are a rare cause of endophthalmitis, and this is the first reported case of endophthalmitis caused by the corynebacterium species Turicella otitidis. As in this case, post-intravitreal injection endophthalmitis may have a bacterial etiology even with delayed presentation. The relatively indolent disease course and excellent response to intravitreal antibiotics is consistent with previous ophthalmic reports regarding other corynebacteria, as well as with otolaryngology and hematology oncology reports addressing Turicella otitidis specifically. This case supports the growing body of evidence for pathogenicity of Turicella otitidis and demonstrates the utility of PCR for diagnosis in small volume aqueous specimens.

Purpose: Rapid identification of virulent pathogens is essential to strengthen the therapeutic strategy of acute endophthalmitis., Objectives: This study sought to compare the contribution of a combination of polymerase chain reaction (PCR)-based tests to culture methods, in patients with postoperative endophthalmitis., Design: Prospective multicenter study diagnostic evaluation., Methods: Setting: university referral centers., Participants: 153 consecutive patients presenting with acute or delayed-onset postoperative endophthalmitis, between 2008 and 2015. There were a total of 284 aqueous humor (AH) and/or vitreous fluid (VF) samples. Outcomes and measurements: microbiological tests of intraocular samples included bacterial culturing of pediatric blood culture bottles; 16SrDNA amplification and sequencing (panbacterial PCR) for detection and identification of all bacterial species; real-time PCR (qPCR) assays targeting the femA or lytA gene for detection of Staphylococcus aureus (S. aureus) or Streptococcus pneumoniae (S. pneumoniae), respectively; and a qPCR assay targeting the tuf gene for detection and quantification of Staphylococcus epidermidis (S. epidermidis)., Results: At the time of admission, the rate of detection of microorganisms by PCR-based tests was not significantly different than that by culturing (38% versus 30% in AH samples [n = 69]; 66% versus 63% in VF samples [n = 82], respectively). In contrast, after 1 intravitreal injection (IVI) of antibiotics, the identification rate by PCR-based tests was higher than that in VF by culturing (62% vs 48%, respectively; n = 94; P = 0.05). Bacteria were identified in 70% of patients, with a predominance of Gram-positive bacteria (93%). Specific qPCR tests targeting S. aureus and S. pneumoniae did not provide additional diagnoses but provided earlier results. The S. epidermidis load in vitreous at the time of patients' admission was higher in cases of final visual acuity (VA) of <20/40 (127,118 ± 125,848 DNA copies/mL) in patients with a VA of ≥20/40 (40350,000 ± 46,912 DNA copies/mL; P = 0.09). No significant changes in S. epidermidis load was found after one IVI., Conclusions: Patients with acute or delayed-onset endophthalmitis should benefit from microbiological identification in vitreous samples by combined analysis using bacterial cultures in pediatric blood culture bottles and panbacterial PCR. The last test was more effective than cultures in vitreous samples collected after an IVI of antibiotics. The qPCR tests targeting S. aureus and S. pneumoniae gave earlier results than culture and panbacterial PCR but did not provide additional diagnoses. As for S. epidermidis infections, determination of bacterial load using the qPCR test targeting the tuf gene could help evaluation of the visual prognosis of patients. Its role in the follow-up of patients after antibiotic treatment needs further investigation., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Objective: To evaluate the role of intraocular fluid analysis as a diagnostic aid for uveitis., Methods: Twenty-eight samples (27 patients including 3 HIV-infected patients) with active (n=24) or non-active (n=4) uveitis were submitted to aqueous (AH; n=12) or vitreous humor (VH) analysis (n=16). All samples were analyzed by quantitative PCR for herpes simplex virus (HSV), varicella zoster virus (VZV), cytomegalovirus (CMV), Epstein-Barr virus (EBV) and Toxoplasma gondii., Results: The positivity of the PCR in AH was 41.7% (5/12), with 50% (2/4) in immunocompetent and 67% (2/3) in HIV+ patients. The positivity of the PCR in VH was 31.2% (5/16), with 13% (1/8) in immunocompetent and 50% (4/8) in immunosuppressed HIV negative patients. The analysis was a determinant in the diagnostic definition in 58% of HA and 50% of VH., Conclusion: Even in posterior uveitis, initial AH analysis may be helpful. A careful formulation of possible clinical diagnosis seems to increase the chance of intraocular sample analysis being meaningful.

Purpose: The aim of this study was to identify the factors associated with positive culture sample in patients with endophthalmitis based on clinical presentation and multimodal intraocular sampling., Design: Retrospective review., Methods: A total of 259 subjects with a diagnosis of endophthalmitis presented to a tertiary ophthalmic referral center between 2006 and 2018. Patient demographics, presenting clinical findings and the results of aqueous and vitreous sampling were analyzed., Results: Mean age was 64.2 (± 22.6) years with 52.9% female. Endophthalmitis followed cataract surgery in 84 eyes (32.4%) and was the most common precipitant; intravitreal injections were the next common cause involving 60 eyes (23.2%). Mean visual acuity on presentation was hand movements with a hypopyon present 134 eyes (52%). In total, 135 cases (52.1%) were culture positive. Aqueous sampling was performed in 112 eyes [culture positive 36 (32.1%)]; vitreous sample in 122 eyes [positive in 56 (45.3%)]. Vitrectomy was performed in 169 eyes with 149 sent for culture [70 (47.0%) positive]. A positive vitrectomy culture was observed in 14 eyes (36.9%) of 38, despite initial treatment with intravitreal antibiotics. Factors associated with positive culture were aqueous tap [odds ratio (OR) 2.06, P = 0.02], vitrectomy (OR 2.86, P = 0.001), and absent red reflex (OR 2.73, P = 0.001)., Conclusions: A multimodal approach to intraocular sampling should be considered in those presenting with endophthalmitis, with both aqueous tap and vitrectomy associated with an increased probability of achieving a positive culture.

Purpose: Infectious endophthalmitis (IE) is a severe complication that can lead to blindness even with treatment. However, the impact of antibiotic resistance and early vitrectomy on its prognosis has scarcely been documented. This study investigated the impact of antibiotic resistance of pathogen and early vitrectomy on the prognosis of IE., Methods: The medical records of 171 patients treated for IE at a tertiary referral center between 2007 and 2016 were retrospectively reviewed and analyzed for etiology, pathogen, drug resistance to vancomycin or third-generation cephalosporins, treatment types and timing, and visual outcomes. Multivariate logistic regression analysis was used to determine significant prognostic factors., Results: Among 171 eyes, 121 (70.8%) eyes developed IE after intraocular surgery (cataract surgery, 46.3%; intraocular injection, 13.2%), 37 (21.6%) eyes developed IE endogenously, and 9 (5.3%) eyes developed IE after trauma. The major causative pathogens were Staphylococcus aureus (9.4%) and Klebsiella pneumoniae (7.0%). In total, 72.6% of the identified pathogens demonstrated antibiotic resistance. Antibiotic resistance was associated with a worse final vision (P = .027). Visual prognosis was better for eyes treated with early vitrectomy combined with intravitreal antimicrobial injections within 24 h of onset than for eyes that received only intravitreal antimicrobial injections before undergoing delayed vitrectomy (P = .003)., Conclusion: Antibiotic resistance of organisms causing IE is one of the most important prognostic factors. Early vitrectomy (i.e., within 24 h) may be helpful for achieving a better visual outcome. Immediate vitrectomy can be recommended, especially in IE cases caused by organisms with resistance to empirically used antibiotics.

Purpose : To analyze the role of polymerase chain reaction (PCR) of ocular fluids in management of tubercular (TB) anterior, intermediate, posterior, and panuveitis. Methods: In Collaborative Ocular Tuberculosis Study (COTS)-1 (25 centers, n  = 962), patients with TB-related uveitis were included. 59 patients undergoing PCR of intraocular fluids (18 females; 53 Asian Indians) were included. Results: 59 (6.13%) of COTS-1 underwent PCR analysis. PCR was positive for Mycobacterium TB in 33 patients (23 males; all Asian Indians). 26 patients were PCR negative (18 males). Eight patients with negative PCR had systemic TB. Anti-TB therapy was given in 18 negative and 31 PCR cases. At 1-year follow-up, five patients with positive PCR (15.15%) and three with negative PCR (11.54%) had persistence/worsening of inflammation. Conclusions : Data from COTS-1 suggest that PCR is not commonly done for diagnosing intraocular TB and positive/negative results may not influence management or treatment outcomes in the real world scenario.

Purpose : To compare genetic testing for microbes in infectious endophthalmitis or uveitis to culture. Methods : This was a retrospective, single-center case series that enrolled patients with clinically suspected endophthalmitis or uveitis of unknown etiology. Aqueous humor or vitreous was collected and sent for routine cultures and genetic testing. Results : In total, 46 patients were enrolled. Genetic testing was positive in 32/46 (70%) cases and culture 6/46 cases (13%). Five of 16 uveitis cases had a final clinical diagnosis of infectious uveitis, and polymerase chain reaction (PCR) was positive in 4/5 cases (80%), versus 0% for culture. In uveitis cases, PCR was 80% sensitive and 82% specific, and culture had 0% sensitivity. The overall sensitivity and specificity of PCR for all cases were 85% and 67%, respectively, compared with 17% and 100% for culture. Conclusion : Genetic assays are inexpensive ($25/case) and more sensitive than culture for identifying intraocular pathogens in endophthalmitis and uveitis.

Purpose: To assess the efficacy of latent tuberculosis (TB) treatment for immunocompetent uveitis patients with a positive T-SPOT.TB test., Methods: This is a consecutive case series of all T-SPOT.TB positive latent TB patients with presumed tuberculous uveitis managed with anti-tuberculous therapy (ATT) from 1 January 2000 to 31 December 2015. Patients with active TB or other known causes of uveitis, immunocompromised states and those followed up < 12 months were excluded. Descriptive statistics and hypothesis testing were performed, with a significance level of p < 0.05 taken., Results: Among the 75 T-SPOT.TB tests performed for uveitis, 14 cases were enrolled. Mycobacterium tuberculosis was isolated in none of the sputum and intraocular samples. Most cases had posterior uveitis (10/14 cases, 71.4%) and/or intermediate uveitis (9/14 cases, 64.3%). Vasculitis was predominantly occlusive. The mean presenting best-corrected visual acuity (BCVA) was 0.18, improving to 0.44 at 6 months (p = 0.03) and 0.40 at 12 months. (p = 0.03). At 1 year, remission of uveitis was achieved in 92.9%, in which none of them recurred at the last follow-up. High-dose systemic steroid was required in 50% of patients. Only 1 patient was steroid dependent at 18 months. The BCVA improvement in patients treated with or without steroid was comparable., Conclusion: In a TB-endemic region with wide Bacille Calmette-Guerin vaccination coverage, ATT for immunocompetent uveitis patients with latent TB identified from T-SPOT.TB test can improve vision, induce long-term steroid-free remission, and prevent recurrence and systemic reactivation of TB in those who require steroid.

Including viral retinitis, endophthalmitis and ocular toxoplasmosis etc, Infectious ocular diseases, which include viral retinitis, endophthalmitis and ocular toxoplasmosis etc, are the common causes leading to blindness. Traditional examinations of serum are of limited help in diagnosing these diseases and are sometimes misleading. Examinations of pathogen DNA, antibodies and cytokines in the intraocular fluids including aqueous humor and vitreous humor can help clinicians get in situ information and hereby confirm diagnosis rapidly. (Chin J Ophthalmol, 2018, 54: 551-556) .

Endogenous bacterial endophthalmitis, also called metastatic endophthalmitis, is a rare bacterial endophthalmitis derived from distant infectious foci via the bloodstream. This infection can potentially cause not only severe visual disturbance, but also loss of the eyeball or death, as most patients are immunocompromised. This retrospective Japanese multicenter study analyzed 32 eyes in 25 definitive cases. Twelve patients (48.0%) had diabetes mellitus. Typical ocular findings were vitreous haze (87.5%), cells in the anterior chambers (62.5%) and retinal infiltrates (50.0%). Elevated body temperature (64.0%), high serum C-reactive protein (96.0%) and leukocytosis (52.0%) were also frequently observed. Culture positivity rates for intraocular fluid were higher in the vitreous (62.5%) versus aqueous humor (28.6%). High positivity rates were also observed for blood (57.1%) and central venous catheters (100%). The most common pathogen was Staphylococcus aureus (10 cases), including methicillin-resistant S. aureus (4 cases). The next most common pathogen was Klebsiella pneumoniae (7 cases), which was highly associated with liver abscess. Compared to a previous 1991 national multicenter study, there has been a fourfold increase in the ratio of S. aureus. Antibiotic susceptibility tests revealed that all Gram-positives were susceptible to vancomycin and all Gram-negatives were susceptible to third-generation cephalosporins, imipenem/cilastatin, gentamycin and levofloxacin. Prognostic factors influencing poor visual outcome included poor initial visual acuity (p < 0.01), K. pneumoniae (p = 0.027) and gram-negative bacteria (p = 0.014) as the causative bacteria. Intravitreal antibiotic injection in combination with vancomycin and ceftazidime may be applicable for use as part of the standard treatment regimen for EBE., (Copyright © 2018 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Polymerase chain reaction (PCR)-based diagnosis of tuberculosis-associated uveitis (TBU) in TB-endemic countries is challenging due to likelihood of latent mycobacterial infection in both immune and non-immune cells. In this study, we investigated normalised quantitative PCR (nqPCR) in ocular fluids (aqueous/vitreous) for diagnosis of TBU in a TB-endemic population. Mycobacterial copy numbers (mpb64 gene) were normalised to host genome copy numbers (RNAse P RNA component H1 [RPPH1] gene) in TBU (n = 16) and control (n = 13) samples (discovery cohort). The mpb64:RPPH1 ratios (normalised value) from each TBU and control sample were tested against the current reference standard i.e. clinically-diagnosed TBU, to generate Receiver Operating Characteristic (ROC) curves. The optimum cut-off value of mpb64:RPPH1 ratio (0.011) for diagnosing TBU was identified from the highest Youden index. This cut-off value was then tested in a different cohort of TBU and controls (validation cohort, 20 cases and 18 controls), where it yielded specificity, sensitivity and diagnostic accuracy of 94.4%, 85.0%, and 89.4% respectively. The above values for conventional quantitative PCR (≥1 copy of mpb64 per reaction) were 61.1%, 90.0%, and 74.3% respectively. Normalisation markedly improved the specificity and diagnostic accuracy of quantitative PCR for diagnosis of TBU., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Advances in genomics have the potential to revolutionize clinical diagnostics. Here, we examine the microbiome of vitreous (intraocular body fluid) from patients who developed endophthalmitis following cataract surgery or intravitreal injection. Endophthalmitis is an inflammation of the intraocular cavity and can lead to a permanent loss of vision. As controls, we included vitreous from endophthalmitis-negative patients, balanced salt solution used during vitrectomy and DNA extraction blanks. We compared two DNA isolation procedures and found that an ultraclean production of reagents appeared to reduce background DNA in these low microbial biomass samples. We created a curated microbial genome database (>5700 genomes) and designed a metagenomics workflow with filtering steps to reduce DNA sequences originating from: (i) human hosts, (ii) ambiguousness/contaminants in public microbial reference genomes and (iii) the environment. Our metagenomic read classification revealed in nearly all cases the same microorganism that was determined in cultivation- and mass spectrometry-based analyses. For some patients, we identified the sequence type of the microorganism and antibiotic resistance genes through analyses of whole genome sequence (WGS) assemblies of isolates and metagenomic assemblies. Together, we conclude that genomics-based analyses of human ocular body fluid specimens can provide actionable information relevant to infectious disease management.

Background: Endophthalmitis is a rare but severe complication following PPV. The incidence of endophthalmitis varies between 20-gauge, 23-gauge, and 25-gauge incisions. The incidence and clinical features of endophthalmitis after 23-gauge PPV in an eye hospital in China was reported in this study., Methods: Data of the eyes that underwent 23-gauge PPV from January 2011 to December 2014 at the Eye Hospital of Wenzhou Medical University was retrospectively collected. All the information was obtained from the electronic medical system. The exclusion criteria included: (1) preoperative diagnosis of endophthalmitis; (2) history of vitrectomy; (3) intraocular surgery within 6 months; (4) history of ocular penetrating trauma; (5) sutures for any of the 3 sclerotomy incisions; (6) patients with cancer, acquired immune deficiency syndrome, or taking drugs that may influence the immune system. The diagnosis of endophthalmitis was based on clinical characteristics and/or culture results from an operative sample., Results: Three thousand nine hundred seventy nine eyes that underwent 23-gauge PPV surgery were included in this study. Among these eyes, 3 eyes developed endophthalmitis after surgery, giving an incidence of 0.075% (3/3979). The period in which endophthalmitis developed ranged from 1 to 5 days post-operation. The visual acuity decreased to hand motions or light perception postoperatively. The culture of aqueous and vitreous of the 2 eyes revealed Staphylococcus epidermidis and enterococcus faecalis respectively, however was negative for the third eye. All 3 eyes had a favorable response to the treatment of vitreous tap and intravitreal antibiotics injection. Two eyes gained visual acuity of 0.05 and 0.5, respectively at the final visit., Conclusions: Endophthalmitis is a rare but sight-threatening complication after 23-gauge pars plana vitrectomy. The peak duration of onset was within 5 days post-operation, with gram positive cocci being the common pathogenic organism.

Purpose: To retrospectively analyze clinical features, laboratory investigations, treatment and visual outcomes in patients with tubercular subretinal abscess., Methods: A total of 12 eyes of 12 patients receiving a diagnosis of subretinal abscess, between 2004 and 2014, were included for this retrospective study., Results: The mean age of the presentation was 29.75 ± 16.72 years (range: 14-62 years) and seven (58.3%) were male. The most common anterior segment presentation was anterior uveitis (75%). The mean BCVA at presentation was 1.62 (in logMAR). A tuberculin skin test was positive in seven patients (58.3%) and five patients had a history of pulmonary tuberculosis. Polymerase chain reaction for the Mycobacterium tuberculosis genome was positive in 6 of 11 eyes from aqueous aspirate (54%) and 4 of 7 eyes from vitreous aspirates (57.14%). All patients were started on systemic steroids and an anti-tubercular regimen. The mean duration of follow-up was 134.28 days. The mean BCVA at final presentation was 1.19 (in logMAR). Chorioretinal scar was the most common finding after resolution., Conclusions: Subretinal abscess is a rare manifestation of ocular tuberculosis. A high degree of suspicion and timely management of the condition can prevent loss of vision.

Introduction: The cause of Seasonal hyperacute panuveitis (SHAPU) also known as seasonal endophthalmitis is not yet confirmed. So far, bacteria have been isolated from the vitreous of three cases, VZV from the aqueous of one case and anellovirus from the vitreous of 30 cases of SHAPU. Its connection with moth is still assumed but not proven yet., Objective: To study the clinical and microbiological features of SHAPU. To find out indirect evidence of it connection with moths., Method: Prospective longitudinal study of all the cases of SHAPU observed at Tilganga Institute of Ophthalmology in 2009 and 2010. A detailed ocular examination was performed on each case and vitreous fluid was removed from almost all cases for microbiological study., Result: 66 cases of SHAPU were identified. Most were children. Twenty-three percent reported a recent history of white moth contact. Fine brown hairs, were present in 5 eyes. Bacteria were isolated from the vitreous of 9 eyes, PCR testing of the vitreous of all 48 cases showed no evidence of HSV, VZV or CMV DNA. Twenty of 34 cases (58.8%) who presented within 7 days of onset of symptoms had a final visual acuity ≥ 6/60 whereas 4/25 cases (16.0%) who presented after 7 days had final visual acuity ≥ 6/60 (p=0.001)., Conclusion: Herpes group of virus is the less likely cause of SHAPU. Bacteria are found in more cases. Early presentation is associated with a better visual outcome. Role of moth in its pathogenesis is still suspected., (© NEPjOPH.)

Bacterial contamination of the anterior chamber during cataract surgery is one of the main responsible for endophthalmitis postoperative. Phacoemulsification is a less invasive technique for cataract treatment, although it does not exclude the possibility of contamination. In this study, bacterial contaminants of aqueous humor collected pre- and post-phacoemulsification with intraocular lens implantation (IOL) of twenty dogs were identified. As the conjunctival microbiota constitute a significant source of anterior chamber contamination, bacterial isolates from aqueous humor were genetically compared with those present in the conjunctival surface of the patients. Three dogs presented bacterial growth in both aqueous humor and conjunctival surface samples. Bacterial isolates from these samples were grouped according to their genetic profiles by repetitive-element PCR (rep-PCR) and their representatives were identified by 16S rRNA sequencing. Isolates from conjunctival surface were identified as Enterobacter spp., Staphylococcus spp. and S. aureus; and from aqueous humor samples as Enterobacter spp., Pantoea spp., Streptococcus spp. and Staphylococcus spp., respectively in decreasing order of prevalence. According to the rep-PCR analysis, 16.6% of Enterobacter spp. isolates from conjunctival surface were genetically similar to those from aqueous humor. The rest of isolates encountered in aqueous humor were genetically distinct from those of conjunctival surface. The significant genetic diversity of bacterial isolates found in the aqueous humor samples after surgery denoted the possibility of anterior chamber contamination during phacoemulsification by bacteria not only from conjunctival surface but also from different sources related to surgical environment., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Purpose: The host immune reaction during endophthalmitis, studied in particular through the intraocular cytokine network, is essential for the comprehension of the disease and the development of new therapies. Therefore, the purpose of this study was to elucidate the cytokine composition of aqueous humor during endophthalmitis., Methods: In a multicenter case-control study, forty-nine patients with postoperative endophthalmitis and 60 controls (cataract surgery) were included. Visual acuity, local inflammatory grading, medical history and intraocular levels of 27 cytokines and chemokines (measured by multiplex immunoassay) were recorded., Results: During endophtalmitis, an increase in total cytokines was observed. The raise of Th1 cytokines was particularly noticeable. Chemokines, such as IL-8, MIP-1 β, MCP-1, G-CSF and IP-10, also increased. Pearson's correlation analyses showed a poor visual prognosis with high levels of IL-8, MCP-1 and VEGF and a low level of IL-10 at admission., Conclusion: An increase in inflammatory cytokines is noticeable during endophthalmitis, with a particular emphasis on IL-8, MCP-1 and VEGF. Targeted anti-inflammatory and anti-VEGF treatments may be of interest in the future.

Purpose: To analyze the clinical features, course, management, and outcomes of tubercular (TB) uveitis in the pediatric population and assess the response to anti-tubercular therapy (ATT)., Methods: Hospital records of children (≤16 years) from a large tertiary-care institute between January 2001 and December 2015 were reviewed., Results: A total of 32 children (mean age: 10.7 ± 4.27 years; range 2-16) were diagnosed with TB-associated uveitis. The most common presentation was posterior uveitis (n = 14, 43.75%) and panuveitis (n = 14, 43.75%), followed by intermediate uveitis (n = 2, 6.25%) and anterior uveitis (n = 2, 6.25%); 14 children had probable intraocular tuberculosis (IOTB) (43.75%) and 17 (53.13%) had possible IOTB. Despite ATT and corticosteroids, 29.63% patients showed suboptimal response or worsening of disease requiring additional immunosuppression., Conclusions: TB is an important cause of pediatric uveitis in endemic countries. The manifestations of the disease resemble adult TB-related uveitis. However, higher inflammatory response in children may require more aggressive therapy with corticosteroids/immunosuppression.

Purpose: To compare the characteristics, visual outcome, and prognostic factors of patients with endogenous endophthalmitis and to determine the association of endophthalmitis with Klebsiella pneumoniae infection., Methods: We retrospectively analyzed records of patients diagnosed with endogenous endophthalmitis from January 2002 to August 2013., Results: A total of 86 patients were diagnosed with endogenous endophthalmitis; 48 patients were infected with K. pneumoniae, 28 patients were infected with other pathogens, and 10 were culture-negative. Diabetes mellitus was more prevalent among patients infected with K. pneumoniae. Liver abscess and urinary tract infection were the leading sources of infection in patients with and without a K. pneumoniae infection, respectively. In patients with endogenous endophthalmitis, poor initial vision (p<0.001) and K. pneumoniae infection (p = 0.048) were significantly associated with a poor visual outcome., Conclusions: Poor initial vision and K. pneumoniae infection were significantly associated with poorer visual outcome for patients with endogenous endophthalmitis.

Purpose: To describe a case of Coccidioides endophthalmitis that resulted in a favorable visual outcome after a combined medical and surgical approach., Methods: A 33-year-old previously healthy woman was referred for evaluation of dyspnea and left-sided vision loss, which began 3 months before, after a trip to Nevada. She was found to have a pulmonary cavitary lesion and fluffy white material in the anterior chamber. An aqueous and vitreous paracentesis grew Coccidioides species. She was managed medically with a total of 7 weekly intravitreal injections of amphotericin B and intravenous liposomal amphotericin B followed by transition to oral posaconazole. Seven months after presentation, to ensure ocular sterilization and to clear the visual axis, she underwent temporary keratoprosthesis implantation, anterior segment reconstruction, removal of a cyclitic membrane and the crystalline lens, pars plana vitrectomy, placement of a pars plana Ahmed drainage device, and penetrating keratoplasty., Results: After surgical intervention and with maintenance posaconazole therapy, the patient had resolution of her dyspnea and improved uncorrected (aphakic) vision with a clear corneal graft, quiet anterior chamber, and normal optic nerve and retina., Conclusions: A combined medical and surgical approach resulted in a favorable visual outcome and avoided the need for enucleation.

Background: Equine recurrent uveitis (ERU) is a common cause of ocular pain and blindness in horses. Leptospira spp. have been commonly implicated in the pathophysiology of ERU in mainland Europe and the USA. No recent studies have been carried out in the UK, but Leptospira is reported not to be a major factor in the aetiology of ERU in the UK., Objectives: To establish the prevalence of Leptospira-associated ERU in the UK and to identify the serovars involved in these cases; to compare serum vs. aqueous humour antibody levels in cases and controls in order to confirm the diagnosis of Leptospira-associated ERU, and to assess the usefulness of serology alone as a confirmatory test for Leptospira-associated ERU in the UK., Study Design: Case-control study., Methods: Eyes enucleated for clinical reasons in ERU-affected horses were collected. Blood and aqueous humour were obtained to determine antibody levels against a variety of Leptospira serovars and C-values (aqueous humour value/serum value) were calculated. In addition, eyes, blood and aqueous humour were obtained from control cases for comparison. Histopathology was performed in all eyes to confirm uveitis in each case. Differences in seroprevalences between ERU and control cases and between Leptospira- and non-Leptospira-associated ERU cases were calculated., Results: A total of 30 ERU and 43 control eyes were analysed. Of the ERU eyes, only two had a C-value of >4 (prevalence of Leptospira-associated uveitis: 6.7%). Serovars hardjo and javanica were detected. There was no difference in seroprevalence between horses with uveitis and control cases (65.5% and 41.9%, respectively; P = 0.11) or between Leptospira- and non-Leptospira-associated uveitis cases (100% and 63.0%, respectively; P = 0.52)., Main Limitations: The study was limited by low case numbers. Eyes were presented at different stages of disease. The only test used to detect Leptospira was the microscopic agglutination test., Conclusions: Leptospira-associated ERU is uncommon in the UK. Serology alone may not help to definitively diagnose Leptospira-associated uveitis in this country., (© 2017 The Authors. Equine Veterinary Journal published by John Wiley & Sons Ltd on behalf of EVJ Ltd.)

Purpose: To report the clinical presentation, microbiologic spectrum, and visual outcomes associated with acute-onset infectious endophthalmitis undergoing therapeutic pars plana vitrectomy., Methods: Multicenter interventional retrospective noncomparative consecutive case series. Billing records were reviewed to identify all charts for patients undergoing pars plana vitrectomy within 14 days of diagnosis of acute-onset infectious endophthalmitis over a 4-year period at 5 large tertiary referral retina practices. Statistical analysis was performed to assess for factors associated with visual outcomes., Results: Seventy patients were identified. The most common clinical setting was postcataract surgery (n = 20). Only 3 patients (4.3%) presented with 20/400 or better visual acuity (VA). Although most of the patients initially underwent vitreous tap and intravitreal antibiotic injection (n = 47, 67.1%), all patients eventually underwent pars plana vitrectomy within 14 days of presentation with 68.5% (48/70) of patients undergoing pars plana vitrectomy within 48 hours of presentation. Positive intraocular cultures were obtained in 56 patients (80%). The most common identified organism was Streptococcus sp (n = 19). Visual acuity at last follow-up was 20/400 or better in 19 patients (27.1%). Three patients underwent evisceration or enucleation (4.3%). Last recorded postoperative VA (mean LogMAR 1.99 ± 0.94, Snellen VA equivalent finger count) improved from presenting VA (mean LogMAR 2.37 ± 0.38, Snellen VA hand motions) (P ≤ 0.001). There was no statistically significant correlation between the underlying etiology or the timing of surgery with this VA outcome., Conclusion: Although less than one-third of patients achieved 20/400 or better VA, this VA often improved significantly from presenting VA.

Bacterial endophthalmitis is a potentially blinding intraocular infection. The bacterium Bacillus cereus causes a devastating form of this disease which progresses rapidly, resulting in significant inflammation and loss of vision within a few days. The outer surface of B. cereus incites the intraocular inflammatory response, likely through interactions with innate immune receptors such as TLRs. This study analyzed the role of B. cereus pili, adhesion appendages located on the bacterial surface, in experimental endophthalmitis. To test the hypothesis that the presence of pili contributed to intraocular inflammation and virulence, we analyzed the progress of experimental endophthalmitis in mouse eyes infected with wild type B. cereus (ATCC 14579) or its isogenic pilus-deficient mutant (ΔbcpA-srtD-bcpB or ΔPil). One hundred CFU were injected into the mid-vitreous of one eye of each mouse. Infections were analyzed by quantifying intraocular bacilli and retinal function loss, and by histology from 0 to 12 h postinfection. In vitro growth and hemolytic phenotypes of the infecting strains were also compared. There was no difference in hemolytic activity (1:8 titer), motility, or in vitro growth (p > 0.05, every 2 h, 0-18 h) between wild type B. cereus and the ΔPil mutant. However, infected eyes contained greater numbers of wild type B. cereus than ΔPil during the infection course (p ≤ 0.05, 3-12 h). Eyes infected with wild type B. cereus experienced greater losses in retinal function than eyes infected with the ΔPil mutant, but the differences were not always significant. Eyes infected with ΔPil or wild type B. cereus achieved similar degrees of severe inflammation. The results indicated that the intraocular growth of pilus-deficient B. cereus may have been better controlled, leading to a trend of greater retinal function in eyes infected with the pilus-deficient strain. Although this difference was not enough to significantly alter the severity of the inflammatory response, these results suggest a potential role for pili in protecting B. cereus from clearance during the early stages of endophthalmitis, which is a newly described virulence mechanism for this organism and this infection., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Purpose: To assess aqueous humor antimicrobial activity in vitro after topical 0.5% chloramphenicol application., Materials and Methods: This investigation included 63 eyes from 65 cataract surgery patients. The study group of 48 eyes received preoperatively four topical applications of 0.5% chloramphenicol. The control group of 15 eyes was given no topical applications. Aqueous humor samples were collected for in vitro antimicrobial analysis using Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, and Pasteurella multocida organisms by means of disk diffusion test., Results: No inhibition halo was observed around all aqueous humor samples from all chloramphenicol-treated patients, irrespective of the sample quantity added to the paper disks, with no significant difference from aqueous humor from untreated control patients., Conclusions: Aqueous humor displayed no bactericidal effect against any of the microorganisms evaluated after topical 0.5% chloramphenicol application.

Purpose: To compare aqueous humor concentrations of topically applied moxifloxacin 0.5% ophthalmic solution alone or in combination with dexamethasone 0.1% and to correlate these concentrations with the minimum inhibitory concentrations (MICs) for common endophthalmitis-causing organisms., Methods: Sixty-eight patients undergoing routine phacoemulsification with intraocular lens implantation received either moxifloxacin 0.5% alone or moxifloxacin 0.5% combined with dexamethasone. For both groups, 1 drop of the test solution was instilled 4 times daily 1 day preoperatively and 1 drop 1 h preoperatively. An aqueous humor sample obtained immediately before paracentesis was submitted to high-performance liquid chromatography-tandem mass spectrometry to determine the moxifloxacin concentration., Results: The mean concentrations of moxifloxacin were 986.6 ng/mL in the moxifloxacin with dexamethasone group and 741.3 ng/mL in the moxifloxacin group (P = 0.13). Moxifloxacin concentrations of all samples exceeded the MICs for Staphylococcus epidermidis, S. aureus, and Streptococcus pneumoniae. All samples in the moxifloxacin with dexamethasone group and 94% in the moxifloxacin group achieved the MIC for Enterococcus species. For quinolone-resistant S. aureus, the MIC was achieved in 29% in the moxifloxacin with dexamethasone group and 9% in the moxifloxacin group (P = 0.06)., Conclusion: Aqueous humor moxifloxacin concentrations were higher when topically administrated in combination with dexamethasone compared to the moxifloxacin alone. However, this difference was not statistically significant. Nevertheless, the MICs of the most common pathogens associated with endophthalmitis were exceeded in both study groups.

Aims: The aim of this study was to detect Mycobacterium tuberculosis (MTB) DNA with polymerase chain reaction (PCR) in aqueous or vitreous samples of patients suffering from choroiditis presumed to be infectious origin., Settings and Design: Hospital-based, retrospective case-control study., Subjects and Methods: In all, forty eyes of forty patients with choroiditis divided into two groups - Group A (serpiginous-like choroiditis, ampiginous choroiditis, multifocal choroiditis) and Group B (choroidal abscess, miliary tuberculosis (TB), choroidal tubercle) were analyzed retrospectively. In 27 controls (patients without uveitis undergoing phacoemulsification), anterior chamber aspirate was done and sample subjected to real-time PCR. Patients underwent nested PCR for MTB using IS6110 and MPB64 primers from aqueous (n = 39) or vitreous (n = 1). All patients underwent detailed ophthalmological examination by slit-lamp biomicroscopy, fundus examination by indirect ophthalmoscopy, and fundus photograph and fundus fluorescein angiography if required., Statistical Analysis: Positive results of PCR for MTB within the group and between two groups were statistically analyzed using Chi-square test., Results: There were 25 males and 15 females. Mean age at presentation was 34.66 years (range, 14-62). PCR positivity rates were 41.3% (n = 12/29) and 81.82% (n = 9/11) in Groups A and B, respectively. No controls had PCR-positive result. Comparison of PCR positivity rates showed statistically significant difference between Groups A and B (P = 0.028). Systemic TB was detected in 57.14% (n = 12/21) of all PCR-positive cases (Group A - 33.3%, n = 4/12; Group B - 88.9%, n = 8/9). Systemic antitubercular treatment (ATT) for 9 months and oral steroids were successful in resolution of choroiditis in all PCR-positive patients (n = 21) without disease recurrence., Conclusions: Eyes with choroiditis of suspected/presumed tubercular origin should be subjected to PCR for diagnosis of TB and subjected to ATT for prevention of recurrences.

Purpose: Postoperative endophthalmitis is a potentially sight-threatening complication of cataract surgery. However, the pathophysiological mechanisms are not completely understood. We sought to study and evaluate the intraocular environment (aqueous and vitreous humors), the capsular tissue, and the intraocular lens (IOL) surfaces of normal eyes after long-term uncomplicated cataract surgery., Design: Experimental laboratory investigation., Methods: We studied 69 eyes donated for transplantation that had previously undergone cataract surgery with posterior chamber IOL implantation and that had no recorded clinical history of postoperative inflammation. We assessed the intraocular environment (DNA traces and biofilm formation) by microbiological evaluation of intraocular fluids using conventional microbiology and molecular techniques, including assessment for the presence of microbes (biofilm formation) on the IOL surface by scanning electron microscopy and ultrastructural capsular remnants by transmission electron microscopy., Results: Isolated or aggregated cocci were probable in 18.8% of IOL optic surfaces (n = 13) studied by scanning electron microscopy, suggesting the presence of bacterial biofilm. In 3 intraocular fluid samples for IOLs with biofilm, we identified 16S rDNA by polymerase chain reaction and sequencing. No microbial contamination was found in intraocular fluids by conventional microbiological methods., Conclusions: Our data suggest the possibility of bacterial biofilm formation on the optic surface of IOLs in normal eyes after long-term uncomplicated cataract surgery even in the absence of clinical or subclinical symptoms., (Copyright © 2016 Elsevier Inc. All rights reserved.)

A nocardioform strain IFM 11456T was isolated from the aqueous humor from a patient with endophthalmitis and was characterized to its taxonomic position. IFM 11456T contained arabinose, galactose and meso-diaminopimelic acid in whole-cell hydrolysates and mycolic acids that co-migrated with those from the type strain of Nocardiaasteroides. The acyl type of muramic acid was N-glycolyl. The diagnostic polar lipids were phosphatidylethanolamine, diphosphatidylglycerol and two unidentified glycolipids and the predominant menaquinone was MK-8(H4, ω-cycl.). These characteristics are typical of members of the genus Nocardia. Results of phylogenetic analyses based on 16S rRNA gene sequences indicated that the isolate represented a novel species of the genus Nocardia and was most closely related to the type strains of Nocardia mikamii JCM 15508T (98.1 %) and Nocardiaaobensis IFM 0372T (98.1 %). However, analysis of partial gyrB sequences showed that strain IFM 11456T had 90.2 % similarity to Nocardia concava IFM 0354T and 90 % to Nocardianiigatensis IFM 0330T. The DNA-DNA relatedness values for strain IFM 11456T compared with N. mikamii JCM 15508T, N. aobensisIFM 0372T and N. concava IFM 0354T ranged from 24.4 to 39.9 %. Phenotypic characteristics that differentiated IFM 11456T from phylogenetically related species were growth at 45 °C, utilization of citrate and growth with inositol as a sole carbon source. On the basis of this polyphasic study, the isolate represents a novel species within the genus Nocardia, for which the name Nocardia shinanonensis sp. nov. is proposed. The type strain is IFM 11456T (=NBRC 109590T=TBRC 5149T).