Microbial keratitis in Sao Paulo, Brazil: a 10-year review of laboratory results, epidemiological features, and risk factors / Ceratite microbiana em São Paulo, Brasil: uma revisão de 10 anos dos resultados de laboratório, características epidemiológicas e fatores de risco

ABSTRACT Purpose: To study epidemiological data, laboratory results, and risk factors associated with microbial keratitis. Methods: We conducted a retrospective study of corneal sample cultures from patients with microbial keratitis from January 2010 to December 2019. Results were analyzed according to the etiological diagnosis of bacterial, mycotic, or parasitic infection and were associated with related risk factors. Results: We analyzed 4810 corneal samples from 4047 patients (mean age 47.79 ± 20.68 years; male 53.27%). The prevalence of bacterial, fungal, and Acanthamoeba infections were 69.80%, 7.31%, and 3.51%, respectively. The most frequently isolated bacteria were coagulase-negative Staphylococcus (CoNS) (45.14%), S. aureus (10.02%), Pseudomonas spp. (8.80%), and Corynebacterium spp. (6.21%). Among CoNS, the main agent was S. epidermidis (n=665). For mycotic keratitis, Fusarium spp. (35.42%) and Candida parapsilosis (16.07%) were the most common agents among filamentous and yeasts isolates, respectively. Contact lens use was associated with a positive culture for Acanthamoeba spp. (OR = 19.04; p < 0.001) and Pseudomonas spp. (OR = 3.20; p < 0.001). Previous ocular trauma was associated with positive fungal cultures (OR = 1.80; p = 0.007), while older age was associated with positive bacterial culture (OR = 1.76; p = 0.001). Conclusions: Our findings demonstrated a higher positivity of corneal sample cultures for bacteria. Among those, CoNS was the most frequently identified, with S. epidermidis as the main agent. In fungal keratitis, Fusarium spp. was the most commonly isolated. Contact lens wearers had higher risks of positive cultures for Acanthamoeba spp. and Pseudomonas spp. Ocular trauma increased the risk of fungal infection, while older age increased the risk of bacterial infection.

RESUMO Objetivo: Estudar os dados epidemiológicos, resultados laboratoriais e fatores de risco associados às ceratites infecciosas. Métodos: Estudo retrospectivo das amostras de cultura de córnea em pacientes com ceratites infecciosas entre Janeiro/2010 a Dezembro/2019. Os resultados foram analisados de acordo com o diagnóstico etiológico de infecção bacteriana, fúngica ou parasitária e correlacionado com os fatores de risco relacionados. Resultados: Quatro mil, oitocentas e dez amostras corneanas de 4047 pacientes (média de idade de 47,79 ± 20,68 anos; homens em sua maioria (53,7%) foram incluídas. A prevalência de infecções por bactéria, fungo e Acanthamoeba foram de 69.80%, 7,31%, and 3,51%, respectivamente. A maioria das bactérias mais frequentemente isoladas foram Staphylococcus coagulase-negativo (CoNS) (45,14%), S. aureus (10,02%), Pseudomonas spp. (8,80%), e Corynebacterium spp. (6,21%). Dentre CoNS, o principal agente foi S. epidermidis (n = 665). Nas ceratites fúngicas, Fusarium spp. (35,42%) e Candida parapsilosis (16,07%) foram os agentes mais comuns entre os filamentosos e leveduriformes, respectivamente. O uso de lentes de contato foi associado à cultura positiva para Acanthamoeba spp. (OR = 19,04; p < 0,001) e Pseudomonas spp (OR = 3,20; p < 0,001). Trauma ocular prévio foi associado a culturas positivas para fungo (OR = 1,80; p = 0,007), e idade avançada foi associada a culturas positivas para bactéria (OR = 1,76; p = 0,001). Conclusões: Nossos achados demonstraram uma maior positividade para bactérias em amostras de cultura corneana. Dentre estas, CoNS foi mais frequentemente identificado, sendo S. epidermidis o principal agente. Nas ceratites fúngicas, Fusarium spp. Foi o mais comumente isolado. O risco de positividade para Acanthamoeba spp. e Pseudomonas spp. foi maior em usuários de lentes de contato. Trauma ocular aumentou o risco de cultura positiva para fungo, ao passo que idade mais avançada aumentou o risco de infecção bacteriana.

Microbial keratitis in Sao Paulo, Brazil: a 10-year review of laboratory results, epidemiological features, and risk factors.

PURPOSE: To study epidemiological data, laboratory results, and risk factors associated with microbial keratitis. METHODS: We conducted a retrospective study of corneal sample cultures from patients with microbial keratitis from January 2010 to December 2019. Results were analyzed according to the etiological diagnosis of bacterial, mycotic, or parasitic infection and were associated with related risk factors. RESULTS: We analyzed 4810 corneal samples from 4047 patients (mean age 47.79 ± 20.68 years; male 53.27%). The prevalence of bacterial, fungal, and Acanthamoeba infections were 69.80%, 7.31%, and 3.51%, respectively. The most frequently isolated bacteria were coagulase-negative Staphylococcus (CoNS) (45.14%), S. aureus (10.02%), Pseudomonas spp. (8.80%), and Corynebacterium spp. (6.21%). Among CoNS, the main agent was S. epidermidis (n=665). For mycotic keratitis, Fusarium spp. (35.42%) and Candida parapsilosis (16.07%) were the most common agents among filamentous and yeasts isolates, respectively. Contact lens use was associated with a positive culture for Acanthamoeba spp. (OR = 19.04; p < 0.001) and Pseudomonas spp. (OR = 3.20; p < 0.001). Previous ocular trauma was associated with positive fungal cultures (OR = 1.80; p = 0.007), while older age was associated with positive bacterial culture (OR = 1.76; p = 0.001). CONCLUSIONS: Our findings demonstrated a higher positivity of corneal sample cultures for bacteria. Among those, CoNS was the most frequently identified, with S. epidermidis as the main agent. In fungal keratitis, Fusarium spp. was the most commonly isolated. Contact lens wearers had higher risks of positive cultures for Acanthamoeba spp. and Pseudomonas spp. Ocular trauma increased the risk of fungal infection, while older age increased the risk of bacterial infection.

In Vitro Evaluation of Rose Bengal Photoactivated by Custom-Built Green Light-Emitting Diode Source for Bacteria and Rapidly Growing Mycobacteria Inhibition.

Purpose: In vitro evaluation of rose bengal (RB) photoactivated by our custom-built green light-emitting diode (LED) source for the growth inhibition of bacterial strains and rapidly growing mycobacterial (RGM) isolates in infectious keratitis. Methods: Six corneal clinical bacteria isolates were included in this study: two Gram-positive bacteria (methicillin-resistant Staphylococcus aureus [MRSA] and Staphylococcus epidermidis), two Gram-negative bacteria (Pseudomonas aeruginosa and Serratia marcescens), and two RGM (Mycobacterium chelonae and Mycobacterium abscessus). Microorganisms were cultured and incubated at specific conditions and prepared in suspensions to adjust their concentration to 104 cells/mL. Different treatments were conducted in triplicates: Group I, no treatment; Group II, treated with 0.1% rose bengal alone (exposed to dark for 30 minutes); Group III, exposed to custom green LED for 30 minutes (12.87 J/cm2); and Group IV, treated with 0.1% rose bengal and exposed to custom green LED for 30 minutes. Agar plates were incubated at specific conditions and photographed after growth for pixel analyses. Results: Complete growth inhibition of all bacteria and RGM was observed in Group IV. MRSA and S. epidermidis in Group II also showed complete growth inhibition. Conclusions: The custom-built green LED presented good activity by photoactivating RB and inhibiting micro-organism growth. For the first time, we demonstrated the expressive growth inhibition effect of RB against S. epidermidis, RGM, and S. marcescens. Clinical treatment with RB may offer an alternate adjunct therapy for corneal surface infections. Translational Relevance: Validating in vitro the custom-built green LED encourages the clinical application for the treatment of infectious keratitis.

Candida species causing fungal keratitis: molecular identification, antifungal susceptibility, biofilm formation, and clinical aspects.

The study aimed to evaluate the clinical aspects, molecular identification, biofilm formation, and antifungal susceptibility profile of Candida species isolated from fungal keratitis. Thirteen Candida isolates from 13 patients diagnosed with Candida keratitis were retrieved and grown in pure culture. Species identification was performed by micromorphology analysis and ITS-rDNA sequencing. The broth microdilution method tested the minimum inhibitory concentration (MIC) of four antifungal drugs (fluconazole, amphotericin B, voriconazole, and anidulafungin). The biofilms were cultured and incubated with antifungal drugs for 24 h. The XTT reduction assay measured the biofilm activity. Biofilm MICs were calculated based on a 50% reduction in metabolic activity compared with the activity of the drug-free control. Among isolates, two were C. albicans, 10 were C. parapsilosis (sensu stricto), and one was C. orthopsilosis. All isolates were classified as susceptible or intermediate to all four antifungal drugs. Four isolates were very low biofilm producers (30%). Nine isolates were biofilm producers, and all biofilm samples were unsusceptible to all drugs tested. Previous ocular surgery was the most common underlying condition for fungal keratitis (84.6%), and C. parapsilosis was the most frequent Candida species (76.9%). Four patients (30.7%) needed keratoplasty, whereas two (15.3%) required evisceration. The biofilm formation ability of Candida isolates decreased antifungal susceptibility compared with planktonic cells. Despite in vitro antifungal susceptibility, almost half of the patients were unresponsive to clinical treatment and needed surgery.

Bilateral interstitial keratitis in a patient with presumed brain tuberculoma / Ceratite intersticial bilateral em um paciente com tuberculoma cerebral presumido

ABSTRACT Interstitial keratitis is an inflammation of the corneal stroma without epithelium or endothelium involvement. The underlying causes are mostly infectious or immune mediated. Brazil has one of the highest incidence rates of tuberculosis in the world. Tuberculosis is considered one of the causes of interstitial keratitis. Malnutrition and anemia are risk factors of the disseminated disease. This is a case report of a 10-year-old child who presented with decreased visual acuity and a clinical diagnosis of bilateral interstitial keratitis and sclero-uveitis. The patient had been treated with topical steroids with partial improvement. Examinations revealed severe iron deficiency anemia, negative serologies for human immunodeficiency virus and syphilis, positivity for cytomegalovirus- and herpes simplex-specific IgG, and purified protein derivative of 17 mm. During the follow-up, the patient presented with tonic-clonic seizures, and magnetic resonance imaging findings suggested a central nervous system tuberculoma. Interstitial keratitis improvement was observed after specific tuberculosis treatment. This is the first case report describing the association of interstitial keratitis and central nervous system tuberculoma.

RESUMO A ceratite intersticial é uma inflamação do estroma corneano sem envolvimento epitelial ou endotelial causada principalmente por doenças infecciosas e imunomediadas. O Brasil tem altas taxas de tuberculose que deve ser lembrada como causa de ceratite intersticial. Desnutrição e anemia são fatores de risco da forma disseminada da tuberculose. Este é um relato de uma criança de 10 anos com redução de acuidade visual e diagnóstico clínico de ceratite intersticial bilateral e esclerouveíte. O paciente obteve melhora parcial da ceratite com corticoide tópico. Exames laboratoriais mostraram anemia ferropriva grave, sorologias negativas para HIV e sífilis; IgM negativo e IgG positivo para citomegalovírus e herpes simplex e PPD positivo (17 mm). Ele evoluiu com crises tônico-clônicas e a ressonância nuclear magnética revelou tuberculoma do sistema nervoso central. A melhora da ceratite intersticial foi observada após tratamento para tuberculose. Este é o primeiro caso que descreve a associação de ceratite intersticial e tuberculoma do sistema nervoso central.

Bilateral interstitial keratitis in a patient with presumed brain tuberculoma.

Interstitial keratitis is an inflammation of the corneal stroma without epithelium or endothelium involvement. The underlying causes are mostly infectious or immune mediated. Brazil has one of the highest incidence rates of tuberculosis in the world. Tuberculosis is considered one of the causes of interstitial keratitis. Malnutrition and anemia are risk factors of the disseminated disease. This is a case report of a 10-year-old child who presented with decreased visual acuity and a clinical diagnosis of bilateral interstitial keratitis and sclero-uveitis. The patient had been treated with topical steroids with partial improvement. Examinations revealed severe iron deficiency anemia, negative serologies for human immunodeficiency virus and syphilis, positivity for cytomegalovirus- and herpes simplex-specific IgG, and purified protein derivative of 17 mm. During the follow-up, the patient presented with tonic-clonic seizures, and magnetic resonance imaging findings suggested a central nervous system tuberculoma. Interstitial keratitis improvement was observed after specific tuberculosis treatment. This is the first case report describing the association of interstitial keratitis and central nervous system tuberculoma.

Corneal ectasia detection by epithelial pattern standard deviation from OCT.

PURPOSE: To test the ability of the corneal epithelial pattern standard deviation (PSD) to distinguish between normal and cases with corneal ectatic condition. SETTING: Instituto de Olhos Renato Ambrósio, Rio de Janeiro, Brazil. DESIGN: Cross-sectional retrospective study. METHODS: Patients were stratified into 4 groups based on clinical data and corneal tomography. Groups 1 and 2 comprised 1 eye randomly selected from 105 patients with normal corneas (N) and 86 patients with bilateral keratoconus (KC). Groups 3 and 4, respectively, comprised 11 ectatic eyes with no surgical treatment for KC (very asymmetric ectasia [VAE]-E) from patients whose fellow eyes (61) presented with normal topographic patterns (VAE-NT). Corneas were scanned using an OCT system (RT Vue) and Scheimpflug corneal tomography (Pentacam) and also had biomechanical assessment through the Corvis ST. Corneal epithelial thickness maps were analyzed, and the PSD value was calculated. The area under the receiver operating characteristic curve analysis was used to evaluate the diagnostic accuracy of the indices. RESULTS: A total of 105 normal eyes, 86 keratoconic eyes, and 11 ectatic eyes whose fellow eyes (61) presented normal topographic patterns were evaluated. Epithelial PSD was significantly different across the 4 groups ( P < .0001). The pairwise comparison revealed that the normal group presented significantly lower values than both ectasia groups (KC and VAE-E, P < .0001) and the VAE-NT group ( P = .0008). There was no statistical significant difference between KC and VAE-E ( P = .4284), while they were significantly higher than the VAE-NT group ( P < .0001 and P = .0004). CONCLUSIONS: Epithelial PSD can be used to detect abnormal epithelial thickness patterns. Corneal epithelial thickness changes could be detected accurately in patients with KC, even in the form fruste of the disease.

Late Spontaneous Postradial Keratotomy Corneal Perforation After Scleral Contact Lens Use-Case Report.

PURPOSE: To describe a case of late spontaneous postradial keratotomy corneal perforation after scleral contact lens (SCL) wear for optic correction. SETTING: Tertiary referral center for corneal pathology. DESIGN: Case report. RESULTS: A 64-year-old man presented the consequences of a late radial keratotomy (RK) surgery performed for myopia correction 26 years ago. His ophthalmologic history was a RK in both eyes (BE), previous Lasik surgery in BE and Lasik enhancement in the right eye (RE), and pterygium excision with conjunctival transplantation in RE. To improve visual acuity, SCL were fitted in both eyes. After 8 months of use, on a certain day, when removing the lens from the RE, the patient reported experiencing intense eye pain and reduced visual acuity. On ophthalmologic examination, the RE cornea was perforated in one of the previous RK incisions. An urgent corneal transplant was performed in the RE, followed by cataract surgery in the same eye. CONCLUSION: Corneal instability caused by RK scars and daily manipulation with the SCL use may have led to ocular perforation.

Evaluation of the Pathogenic Potential of Escherichia coli Strains Isolated from Eye Infections.

While primarily Gram-positive bacteria cause bacterial eye infections, several Gram-negative species also pose eye health risks. Currently, few studies have tried to understand the pathogenic mechanisms involved in E. coli eye infections. Therefore, this study aimed to establish the pathogenic potential of E. coli strains isolated from eye infections. Twenty-two strains isolated between 2005 and 2019 from patients with keratitis or conjunctivitis were included and submitted to traditional polymerase chain reactions (PCR) to define their virulence profile, phylogeny, clonal relationship, and sequence type (ST). Phenotypic assays were employed to determine hemolytic activity, antimicrobial susceptibility, and adhesion to human primary corneal epithelial cells (PCS-700-010). The phylogenetic results indicated that groups B2 and ST131 were the most frequent. Twenty-five virulence genes were found among our strains, with ecp, sitA, fimA, and fyuA being the most prevalent. Two strains presented a hemolytic phenotype, and resistance to ciprofloxacin and ertapenem was found in six strains and one strain, respectively. Regarding adherence, all but one strains adhered in vitro to corneal cells. Our results indicate significant genetic and virulence variation among ocular strains and point to an ocular pathogenic potential related to multiple virulence mechanisms.

The Impact of the COVID-19 Pandemic on Corneal Transplantation in Brazil.

PURPOSE: The purpose of this study was to evaluate the impact of the coronavirus disease 2019 pandemic on corneal transplantation (CT) in Brazil. METHODS: Data from patients who underwent CT at the Hospital Oftalmológico de Sorocaba (HOS), Brazil, were analyzed. National and state numbers of keratoplasties, patients added to the CT waiting list, and total patients on the waiting list were also obtained. Baseline prepandemic (from January 1, 2019, to March 31, 2020) data were compared with 2 time frames of the coronavirus disease 2019 pandemic: elective CT suspension period (between April 1, 2020, and September 31, 2020) and after elective CT resumption (between October 1, 2020, and April 30, 2021). RESULTS: Despite elective CT resumption after the moratorium, the monthly CT rates did not return to baseline at HOS (-14.7%, P = 0.007), São Paulo state (-19.1%, P = 0.001), or Brazil (-30.1%, P < 0.001). The waiting list increased significantly regionally (P < 0.001) and nationally (P < 0.001). Among optical keratoplasties performed at HOS after resuming elective CTs, the proportion of endothelial keratoplasties declined from 38.2% to 30.0% (P < 0.001), whereas penetrating keratoplasties increased from 33.2% to 39.5% (P < 0.001) when comparing with prepandemic data. CONCLUSIONS: Keratoplasty numbers dropped significantly locally, regionally, and nationally. Hence, the CT waiting lists had a progressive increase, with significant long-term implications. An estimated increment on monthly CT rates of approximately 34% in São Paulo state, and 91% in Brazil, is required for the CT waiting list to get back to prepandemic numbers over the next 2 years.

Trends in Corneal Transplantation in a Tertiary Hospital in Brazil.

PURPOSE: The purpose of this study was to analyze the evolving trends of surgical techniques and indications of corneal transplantation (CT) at a tertiary hospital in Brazil. METHODS: The medical records of all patients who underwent CT at the Hospital Oftalmológico de Sorocaba (Sorocaba Eye Hospital) from the Banco de Olhos de Sorocaba (Sorocaba Eye Bank) group in Sorocaba, Brazil, from January 1, 2012, to December 31, 2019, were analyzed. Data regarding age, sex, transplant indication, and surgical technique were collected. RESULTS: A total of 16,250 CTs were performed. There was a statistically significant decreasing trend of keratoconus-related CT ( P < 0.0001), with rates dropping from 41.7% among all CTs in 2012 to 25.5% in 2019. Penetrating keratoplasty, anterior lamellar keratoplasty, and endothelial keratoplasty (EK) accounted for 59.3%, 27.1%, and 7.8% of the CTs performed in 2012 and 33.3%, 16.4%, and 39.9% in 2019, respectively. A statistically significant decreasing trend was observed for penetrating keratoplasty ( P < 0.0001) and anterior lamellar keratoplasty ( P < 0.0001), whereas EK showed a statistically significant increasing trend during the period ( P < 0.0001). Among EKs, Descemet membrane EK increased statistically significantly from 12.8% in 2012 to 74.4% in 2019 ( P < 0.0001). CONCLUSIONS: This study shows relevant evolving trends in indications and preferred CT techniques in a tertiary hospital in Brazil.

Impact of COVID-19 on corneal esthesiometry.

Individuals with suspected COVID-19 symptoms (n=202) were tested using nasopharyngeal RT-qPCR. All individuals underwent corneal esthesiometry measurements using the Cochet-Bonnet esthesiometer during their first visit; 50 participants underwent an additional measurement at a mean interval of 32.5 (17.8) days. COVID-19 was confirmed in 101 subjects (50%) using nasopharyngeal PCR. The mean time from symptom onset to disease diagnosis and corneal esthesiometry was 4.2 (2.1) days. Mean corneal esthesiometry findings based on the positive and negative PCR status indicated no statistical difference. This study demonstrated that COVID-19 had no effect on corneal esthesiometry in individuals with acute-phase COVID-19.

Three Decades of Contact Lens-associated Microbial Keratitis in a Referral Hospital in São Paulo, Brazil / Ceratite Microbiana Associada a Lentes de contato em um Hospital de Referência em São Paulo, Brasil

ABSTRACT Purpose: The aim of this study was to analyze patient data and the laboratory results of corneal samples collected from patients followed at the Ophthalmology Department, Hospital São Paulo, Brazil over a 30-year period, and correlate the analize with contact lens wearing. Methods: Corneal samples from patients diagnosed clinically with microbial keratitis between January 1987 and December 2016 were included in this study. Cultures that were positive for bacteria, fungi, and Acanthamoeba spp. were analyzed retrospectively. To ascertain if the number of patients with contact-lens-associated microbial keratitis (as a risk factor for microbial infection) changed over time, the analysis was divided into three decades: 1987-1996, 1997-2006, and 2007-2016. Information pertaining to patient gender and age, as well as type of organism isolated, were compared among these periods. Results: The corneal samples of 10.562 patients with a clinical diagnosis of microbial keratitis were included in the study, among which 1.848 cases were related to the use of contact lenses. The results revealed that the frequency of contact-lens-associated microbial keratitis increased over the last two decades. Overall, females had contact-lens-associated microbial keratitis more frequently (59.5%). Patients aged 19-40 years also had contact-lens-associated microbial keratitis more frequently in all study periods. Staphylococcus spp. and Pseudomonas spp. were the most frequent Gram-positive and Gram-negative bacteria, respectively, in the microbial keratitis groups. Among the fungal cases of microbial keratitis, filamentous fungi were the most frequent fungi over the entire study period, with Fusarium spp. being the most frequent fungi in the group with microbial keratitis not associated with contact lens wearing (p<0.001). Samples positive for Acanthamoeba spp. and Pseudomonas spp. were significantly correlated with contact-lens-associated microbial keratitis (p<0.001). Conclusions: Females and young adults aged 19-40 years exhibited the highest frequency of contact-lens-associated microbial keratitis in our study. Staphylococcus spp. and Fusarium spp. were the predominant bacteria and fungi, respectively, isolated from corneal samples. Pseudomonas spp. and Acanthamoeba spp. were significantly correlated with contact-lens-associated microbial keratitis in this study.

RESUMO Objetivo: O objetivo deste estudo foi analisar da dos epidemiológicos de pacientes e resultados laboratoriais para todas as amostras de córnea coletadas de pacientes atendidos no Departamento de Oftalmologia do Hospital São Paulo, Brasil, durante um período de 30 anos e correlacionar com o uso de lentes de contato. Métodos: Amostras de córnea de pacientes com diagnóstico clínico de ceratite microbiana (de janeiro de 1987 a dezembro de 2016) foram incluídas neste estudo. Resultados laboratoriais para culturas positivas para bactérias, fungos e Acanthamoeba spp. foram analisados retrospectivamente. Para verificar se o número de pacientes com ceratite microbiana associada à lente de contato, fator de risco para infecção microbiana, mudou ao longo do tempo, a análise foi dividida em três décadas: 1987-1996, 1997-2006 e 2007-2016. As informações incluindo o sexo do paciente, idade e tipo de organismo isolado foram comparadas entre os períodos. A análise estatística foi realizada no software SAS/STAT 9.3 e SPSS (v20.0). Resultados: Amostras de córnea de 10.562 pacientes com ceratite microbiana foram incluídas no estudo, das quais 1.848 foram relacionadas ao uso de lentes de contato. Os resultados revelaram que a frequência de ceratite microbiana associada à lente de contato aumentou nas últimas duas décadas analisadas. No geral, os homens compreendiam uma proporção maior do grupo ceratite microbiana não associada à lente de contato (CMNLC) (60,3%) e as mulheres eram mais frequentes no grupo ceratite microbiana associada à lente de contato (59,5%). Pacientes com idade entre 19 e 40 anos foram mais frequentemente observados no grupo ceratite microbiana associada à lente de contato em todos os períodos. Staphylococcus spp. foi a bactéria Gram-positiva mais frequentes, enquanto Pseudomonas spp. foi a bactéria Gram-negativa nos grupos ceratite microbiana. Entre os fungos ceratite microbiana, os fungos filamentosos foram os fungos mais frequentes durante todo o período do estudo, com Fusarium spp. sendo o mais frequente no grupo ceratite microbiana não associada à lente de contato. Acanthamoeba spp. e Pseudomonas spp. amostras positivas foram significativamente correlacionadas com ceratite microbiana associada à lente de contato. Conclusões: A maior prevalência de ceratite microbiana associada à lente de contato no nosso estudo foi observada em mulheres e adultos jovens com idade entre 19 e 40 anos. Staphylococcus spp. e Fusarium spp. foram as bactérias e fungos predominantes isolados nas amostras da córnea. Pseudomonas spp. e Acanthamoeba spp. foram significativamente correlacionados a ceratite microbiana associada à lente de contato neste estudo.

Three Decades of Contact Lens-associated Microbial Keratitis in a Referral Hospital in São Paulo, Brazil.

PURPOSE: The aim of this study was to analyze patient data and the laboratory results of corneal samples collected from patients followed at the Ophthalmology Department, Hospital São Paulo, Brazil over a 30-year period, and correlate the analize with contact lens wearing. METHODS: Corneal samples from patients diagnosed clinically with microbial keratitis between January 1987 and December 2016 were included in this study. Cultures that were positive for bacteria, fungi, and Acanthamoeba spp. were analyzed retrospectively. To ascertain if the number of patients with contact-lens-associated microbial keratitis (as a risk factor for microbial infection) changed over time, the analysis was divided into three decades: 1987-1996, 1997-2006, and 2007-2016. Information pertaining to patient gender and age, as well as type of organism isolated, were compared among these periods. RESULTS: The corneal samples of 10.562 patients with a clinical diagnosis of microbial keratitis were included in the study, among which 1.848 cases were related to the use of contact lenses. The results revealed that the frequency of contact-lens-associated microbial keratitis increased over the last two decades. Overall, females had contact-lens-associated microbial keratitis more frequently (59.5%). Patients aged 19-40 years also had contact-lens-associated microbial keratitis more frequently in all study periods. Staphylococcus spp. and Pseudomonas spp. were the most frequent Gram-positive and Gram-negative bacteria, respectively, in the microbial keratitis groups. Among the fungal cases of microbial keratitis, filamentous fungi were the most frequent fungi over the entire study period, with Fusarium spp. being the most frequent fungi in the group with microbial keratitis not associated with contact lens wearing (p<0.001). Samples positive for Acanthamoeba spp. and Pseudomonas spp. were significantly correlated with contact-lens-associated microbial keratitis (p<0.001). CONCLUSIONS: Females and young adults aged 19-40 years exhibited the highest frequency of contact-lens-associated microbial keratitis in our study. Staphylococcus spp. and Fusarium spp. were the predominant bacteria and fungi, respectively, isolated from corneal samples. Pseudomonas spp. and Acanthamoeba spp. were significantly correlated with contact-lens-associated microbial keratitis in this study.

The role of Acanthamoeba spp. in biofilm communities: a systematic review.

Acanthamoeba spp. have always caused disease in immunosuppressed patients, but since 1986, they have become a worldwide public health issue by causing infection in healthy contact lens wearers. Amoebae of the Acanthamoeba genus are broadly distributed in nature, living either freely or as parasites, and are frequently associated with biofilms throughout the environment. These biofilms provide the parasite with protection against external aggression, thus favoring its increased pathogeny. This review aims to assess observational studies on the association between Acanthamoeba spp. and biofilms, opening potential lines of research on this severe ocular infection. A systematic literature search was conducted in May 2020 in the following databases: PubMed Central®/Medline, LILACS, The Cochrane Library, and EMBASE®. The studies were selected following the inclusion and exclusion criteria specifically defined for this review. Electronic research recovered 353 publications in the literature. However, none of the studies met the inclusion criterion of biofilm-producing Acanthamoeba spp., inferring that the parasite does not produce biofilms. Nonetheless, 78 studies were classified as potentially included regarding any association of Acanthamoeba spp. and biofilms. These studies were allocated across six different locations (hospital, aquatic, ophthalmic and dental environments, biofilms produced by bacteria, and other places). Acanthamoeba species use biofilms produced by other microorganisms for their benefit, in addition to them providing protection to and facilitating the dissemination of pathogens residing in them.

Ectatic diseases.

Ectatic corneal disease (ECD) comprises a group of disorders characterized by progressive thinning and subsequent bulging of the corneal structure. Different phenotypes have been recognized, including keratoglobus, pellucid marginal degeneration (PMD), and keratoconus (KC). Keratoconus has been widely investigated throughout the years, but the advent of laser refractive surgery boosted an immediate need for more knowledge and research about ectatic diseases. This article discusses nomenclature of ectatic disease, etiology and pathogenesis, along with treatment options, with special focus ok KC and forme fruste keratoconus.

Characterization and distribution of viridans group streptococci isolated from infectious endophthalmitis and keratitis / Caracterização e distribuição de estreptococos do grupo viridans isolados de endoftalmite infecciosa e ceratite

ABSTRACT Purpose: The aims of this study were to characterize alpha-hemolytic streptococci among isolates from cases of infectious endophthalmitis and keratitis and to determine their distributions. Methods: The sample included 27 and 35 nonduplicated isolates of alpha-hemolytic streptococci recovered from patients with infectious endophthalmitis (2002-2013) and keratitis (2008-2013), respectively. Isolates were identified by the optochin susceptibility and bile solubility tests, using a biochemical identification system. The minimum inhibitory concentration was determined by the broth microdilution method. Molecular identification was performed by analyses of three constitutive genes and the complementary multilocus sequence. The molecular epidemiology of Streptococcus pneumoniae was investigated using multilocus sequence typing, and the presence of the capsular polysaccharide-encoding gene was assessed using conventional polymerase chain reaction. Outcomes were evaluated using the patients' medical records. Results: Phenotypic tests differentiated S. pneumoniae from other alpha-hemolytic streptococci, consistent with later molecular identifications. Streptococcus oralis was significantly prevalent among the endophthalmitis isolates, as was S. pneumoniae in the keratitis isolates. High levels of susceptibility to antibiotics were observed, including vancomycin, cephalosporins, and fluoroquinolones. High genetic variability was detected among the 19 S. pneumoniae strains, with 15 predicted to be encapsulated. The medical records of patients with infectious endophthalmitis were reviewed (n=15/27; 56%), and final visual acuity was assessed in 12 cases (44%). Many patients progressed to a final visual acuity state of "no light perception" (6/12; 50%), "light perception" (3/12; 25%), or "hand motion" (1/12; 8%). The medical records of patients with infectious keratitis were also reviewed (n=24/35; 69%), and final visual acuity was assessed in 18 cases (51%). Similarly, most patients progressed to a final visual acuity state of "no light perception" (6/18; 33%), "light perception" (1/18; 6%), or "hand motion" (6/18; 33%). Overall, the majority of patients progressed to a final visual acuity state of "no light perception" (12/30), "light perception" (4/30), or "hand motion" (7/30). Conclusions: The distribution of alpha-hemolytic streptococci in ocular infections suggested the presence of a species-specific tissue tropism. The prognoses of patients with ocular streptococcal infections were highly unfavorable, and antibiotic resistance did not contribute to the unfavorable clinical progressions and poor outcomes.

RESUMO Objetivo: O objetivo deste estudo foi caracterizar os estreptococos alfa-hemolíticos isolados de endoftalmite infecciosa e ceratite e determinar sua distribuição. Métodos: A amostra incluiu 27 e 35 isolados não-duplicados de estreptococos alfa-hemolíticos recuperados de pacientes com endoftalmite infecciosa (2002-2013) e ceratite (2008-2013), respectivamente. Os isolados foram identificados pelos testes de suscetibilidade à optoquina e bile solubilidade, utilizando um sistema de identificação bioquímica. A concentração inibitória mínima foi determinada pelo método de microdiluição em caldo. A identificação molecular foi realizada pela análise de três genes constitutivos e análise complementar de sequências multilocus. A epidemiologia molecular do Streptococcus pneumoniae foi investigada por tipagem de sequência multilocus, e a presença do gene codificador do polissacarídeo capsular foi avaliada por reação em cadeia da polymerase convencional. Os resultados foram avaliados utilizando os prontuários médicos dos pacientes. Resultados: Os testes fenotípicos diferenciaram S. pneumoniae dos outros estreptococos alpha-hemolíticos, consistentes com identificações moleculares posteriores. S. oralis foi significativamente prevalente entre os isolados de endoftalmite, assim como S. pneumoniae nos isolados de ceratite. Foram observados altos níveis de suscetibilidade a antibióticos, incluindo vancomicina, cefalosporinas e fluoroquinolonas. Alta variabilidade genética foi detectada entre as 19 cepas de S. pneumoniae, com 15 previstas para serem encapsuladas. Os prontuários médicos dos pacientes com endoftalmite infecciosa foram revisados (n=15/27; 56%), e a acuidade visual final foi avaliada em 12 casos (44%). Muitos pacientes evoluiram para um estado final de acuidade visual de "sem percepção luminosa" (6/12; 50%), "percepção luminosa" (3/12; 25%) ou "movimentos de mãos" (1/12; 8%). Também foram revisados os prontuários médicos dos pacientes com ceratite infecciosa (n=24/35; 69%), e a acuidade visual final foi avaliada em 18 casos (51%). Da mesma foram, a maioria dos pacientes evoluiu para um estado final de acuidade visual de "sem percepção luminosa" (6/18; 33%), "percepção luminosa" (1/18; 6%) ou "movimentos de mãos" (6/18; 33%). No geral, a maioria dos pacientes evoluiu para um estado final de acuidade visual de "sem percepção luminosa" (12/30), "percepção luminosa" (4/30) ou "movimentos de mãos" (7/30). Conclusões: A distribuição de estreptococos alfa-hemolíticos nas infecções oculares sugeriu a presença de um tropismo de tecido específico da espécie. Os prognósticos dos pacientes com infeções oculares por estreptococos foram altamente desfavoráveis e a resistência a antibióticos contribuiu não para as progressões clínicas des­favoráveis e os maus resultados.

Fungal keratitis management in a referral cornea center in Brazil / Manejo de ceratites fúngicas em centro de referência no Brasil

Abstract Purpose: To report etiological diagnosis, predisposing risk factors, therapeutic strategies and visual outcome of patients treated at the Department of Ophthalmology of Federal University of São Paulo. Methods: This is a retrospective, descriptive, and observational study from medical and laboratory records of the Department of Ophthalmology of Federal University of São Paulo, including all patients with culture proven fungal keratitis in 5 years, from October 2012 through October 2017. Results: There were 2260 fungi microbiologic test requests. Of these, 140 samples had positive cultures for fungi and sixty-six patients were followed at our clinic. Forty-five patients (68.2%) were men, and the mean age was 48.06 (±17.39) years. Fusarium spp. was the most frequently isolated fungus (32 cases; 48.5%), followed by Candida parapsilosis (12 cases; 18.2%). Thirty-four patients (51.5%) underwent intracameral injection of amphotericin B (5 µg per 0.1 ml). In 11 patients (32.3%), infection was eradicated after intracameral amphotericin B associated to topical antifungal treatment and, in 23 patients (67.7%), therapeutic keratoplasty was needed. No complication related to intracameral amphotericin B injection was observed in this series. Forty-three patients (65.1%) ended up with therapeutic keratoplasty. Three patients (4.5%) evolved to evisceration or enucleation. At the last follow-up visit, 53 patients (80.3%) had visual acuity worse than 20/200. Conclusion: Despite current antifungals drugs and distinct administration strategies, fungal keratitis remains challenging. Delayed antifungal therapy may explain poor clinical outcomes. Intracameral amphotericin B associated to topical antfungal treatment seems to be a safe and helpful alternative for non-responsive fungal keratitis. But it is important to formulate other treatment strategies, hence to improve patients' outcomes, since most patients ended-up with significant visual impairment even after current treatment.

Resumo Objetivo: Descrever diagnósticos etiológicos, fatores de risco, estratégias terapêuticas e resultados visuais de pacientes com ceratite fúngica tratados no Departamento de Oftalmologia da Universidade Federal de São Paulo. Métodos: Trata-se de um estudo retrospectivo, descritivo e observacional, a partir da análise de prontuários médicos e laboratoriais do Departamento de Oftalmologia da Universidade Federal de São Paulo, incluindo todos os pacientes com ceratite fúngica comprovada por cultura no período de outubro de 2012 a outubro de 2017. Resultados: Foram realizadas 2260 solicitações de testes microbiológicos. Destas, 140 amostras apresentaram culturas positivas para fungos, e 66 pacientes foram acompanhados em nosso serviço. Quarenta e cinco pacientes (68,2%) eram do sexo masculino, e a média de idade foi de 48,06 (± 17,39) anos. Fusarium spp. foi o fungo mais freqüentemente isolado (32 casos; 48,5%), seguido por Candida parapsilosis (12 casos; 18,2%). Trinta e quatro pacientes (51,5%) foram submetidos à injeção intracameral de anfotericina B (5 µg por 0,1 ml). Destes, 11 pacientes (32,3%) tiveram a infecção erradicada. Nos outros 23 pacientes (67,7%), o transplante terapêutico foi necessário. Nenhuma complicação relacionada à injeção intracameral de anfotericina B foi observada neste estudo. No total, 43 pacientes (65,1%) evoluíram para transplante terapêutico, e 3 pacientes (4,5%) foram submetidos à evisceração ou enucleação. Cinquenta e três pacientes (80,3%) apresentaram acuidade visual final pior que 20/200. Conclusões: Apesar dos diversos medicamentos antifúngicos atuais e vias de administração, o tratamento das ceratites fúngicas permanece desafiador. O atraso no início do tratamento adequado pode justificar o desfecho clínico desfavorável de grande parte dos pacientes. A injeção intracameral de anfotericina B mostrou-se uma alternativa terapêutica segura para ceratites fúngicas refratárias. Mas outras estratégias de tratamento devem ser formuladas, visando melhorar os resultados visuais dos pacientes.

The Role of Corneal Biomechanics for the Evaluation of Ectasia Patients.

PURPOSE: To review the role of corneal biomechanics for the clinical evaluation of patients with ectatic corneal diseases. METHODS: A total of 1295 eyes were included for analysis in this study. The normal healthy group (group N) included one eye randomly selected from 736 patients with healthy corneas, the keratoconus group (group KC) included one eye randomly selected from 321 patients with keratoconus. The 113 nonoperated ectatic eyes from 125 patients with very asymmetric ectasia (group VAE-E), whose fellow eyes presented relatively normal topography (group VAE-NT), were also included. The parameters from corneal tomography and biomechanics were obtained using the Pentacam HR and Corvis ST (Oculus Optikgeräte GmbH, Wetzlar, Germany). The accuracies of the tested variables for distinguishing all cases (KC, VAE-E, and VAE-NT), for detecting clinical ectasia (KC + VAE-E) and for identifying abnormalities among the VAE-NT, were investigated. A comparison was performed considering the areas under the receiver operating characteristic curve (AUC; DeLong's method). RESULTS: Considering all cases (KC, VAE-E, and VAE-NT), the AUC of the tomographic-biomechanical parameter (TBI) was 0.992, which was statistically higher than all individual parameters (DeLong's; p < 0.05): PRFI- Pentacam Random Forest Index (0.982), BAD-D- Belin -Ambrosio D value (0.959), CBI -corneal biomechanical index (0.91), and IS Abs- Inferior-superior value (0.91). The AUC of the TBI for detecting clinical ectasia (KC + VAE-E) was 0.999, and this was again statistically higher than all parameters (DeLong's; p < 0.05): PRFI (0.996), BAD-D (0.995), CBI (0.949), and IS Abs (0.977). Considering the VAE-NT group, the AUC of the TBI was 0.966, which was also statistically higher than all parameters (DeLong's; p < 0.05): PRFI (0.934), BAD- D (0.834), CBI (0.774), and IS Abs (0.677). CONCLUSIONS: Corneal biomechanical data enhances the evaluation of patients with corneal ectasia and meaningfully adds to the multimodal diagnostic armamentarium. The integration of biomechanical data and corneal tomography with artificial intelligence data augments the sensitivity and specificity for screening and enhancing early diagnosis. Besides, corneal biomechanics may be relevant for determining the prognosis and staging the disease.

Characterization and distribution of viridans group streptococci isolated from infectious endophthalmitis and keratitis.

PURPOSE: The aims of this study were to characterize alpha-hemolytic streptococci among isolates from cases of infectious endophthalmitis and keratitis and to determine their distributions. METHODS: The sample included 27 and 35 nonduplicated isolates of alpha-hemolytic streptococci recovered from patients with infectious endophthalmitis (2002-2013) and keratitis (2008-2013), respectively. Isolates were identified by the optochin susceptibility and bile solubility tests, using a biochemical identification system. The minimum inhibitory concentration was determined by the broth microdilution method. Molecular identification was performed by analyses of three constitutive genes and the complementary multilocus sequence. The molecular epidemiology of Streptococcus pneumoniae was investigated using multilocus sequence typing, and the presence of the capsular polysaccharide-encoding gene was assessed using conventional polymerase chain reaction. Outcomes were evaluated using the patients' medical records. RESULTS: Phenotypic tests differentiated S. pneumoniae from other alpha-hemolytic streptococci, consistent with later molecular identifications. Streptococcus oralis was significantly prevalent among the endophthalmitis isolates, as was S. pneumoniae in the keratitis isolates. High levels of susceptibility to antibiotics were observed, including vancomycin, cephalosporins, and fluoroquinolones. High genetic variability was detected among the 19 S. pneumoniae strains, with 15 predicted to be encapsulated. The medical records of patients with infectious endophthalmitis were reviewed (n=15/27; 56%), and final visual acuity was assessed in 12 cases (44%). Many patients progressed to a final visual acuity state of "no light perception" (6/12; 50%), "light perception" (3/12; 25%), or "hand motion" (1/12; 8%). The medical records of patients with infectious keratitis were also reviewed (n=24/35; 69%), and final visual acuity was assessed in 18 cases (51%). Similarly, most patients progressed to a final visual acuity state of "no light perception" (6/18; 33%), "light perception" (1/18; 6%), or "hand motion" (6/18; 33%). Overall, the majority of patients progressed to a final visual acuity state of "no light perception" (12/30), "light perception" (4/30), or "hand motion" (7/30). CONCLUSIONS: The distribution of alpha-hemolytic streptococci in ocular infections suggested the presence of a species-specific tissue tropism. The prognoses of patients with ocular streptococcal infections were highly unfavorable, and antibiotic resistance did not contribute to the unfavorable clinical progressions and poor outcomes.