Epidemiological profile of infectious keratitis / Perfil epidemiológico da ceratite infecciosa

Abstract Objective: To evaluate the epidemiologic aspects of cornea ulcers in a referred ophthalmology private practice center. Methods: Retrospective study over electronic files of patients treated for corneal ulcers during a period of 7 years by the same physician (RJMF) at Centro de Olhos São Francisco, between june 2007 and june 2014. These patients were evaluated for risk factors such as: trauma causes, co-morbidities and contact lenses use. They were also evaluated by the microorganism found at the smears, treatment and patient's outcome. Patients that didn't return with the smears results or that didn´t have a complete follow-up were excluded from the study. Results: A total of 242 patients were fully treated during this period. 55 patients were excluded and 187 patients were included. 28.88% (54/187) were positive for gram negative bacteria, 27.81% (52/187) for filamentous fungi, 16.04% (29/187) for gram positive bacteria and only 1.07% for acanthamoeba. Twenty-four patients had negative cultures and 21 patients had combined infections (bacteria + fungi or other combination). Corneal collagen Cross-linking was effective on melting arrest in 16 of the 16 patients that we have submitted to this procedure during 2008 and 2009. Among all the patients that had corneal infection and were previously using contact lenses, 81% were infected by pseudomonas or pseudomonas associated with other gram negative bacteria. Filamentous fungi had a strong association with physical trauma. While use of saline solution for contact lenses cleaning had a strong association with gram negative bacteria and biological trauma, a strong association with combined infection. Conclusion: Gram negative bacteria (Pseudomonas) followed by filamentous fungi (Fusarium sp) were the most frequent etiologic agent found in our study. Cross-link was effective on corneal melting arrest. Pseudomonas aeruginosa was the most frequent isolated agent on hydrophilic contact lenses wearers.

Resumo Objetivo: Avaliar os aspectos epidemiológicos de úlceras de córnea em um centro privado de referência oftalmológica. Métodos: Estudo retrospectivo a partir de arquivos eletrônicos de pacientes tratados por úlceras de córnea durante um período de 7 anos pelo mesmo médico (RJMF) no Centro de Olhos São Francisco, entre Junho de 2007 e Junho de 2014. Estes pacientes foram avaliados quanto a fatores de risco, trauma, co-morbidades, uso de lentes de contato, tipo de microrganismo encontrado, tratamento e evolução do doente. Os pacientes que não retornaram com os resultados das culturas ou que não possuíam um seguimento completo foram excluídos do estudo. Resultados: Um total de 242 pacientes foram tratados durante este período. 55 pacientes foram excluídos e 187 foram incluídos no estudo. 28,88% (54/187) foram positivos para as bactérias gram-negativas, 27,81% (52/187) para fungos filamentosos, 16,04% (29/187) para as bactérias gram-positivas e apenas 1,07% para acanthamoeba. Vinte e quatro pacientes tiveram culturas negativas e 21 pacientes tinham infecções combinadas (bactérias fungos + ou outra combinação). O Cross-link foi eficaz em interromper o melting em 16 dos 16 pacientes que se submeteram a este procedimento durante 2008 e 2009. Entre todos os pacientes que tiveram infecção de córnea e usavam previamente lentes de contato, 81% foram infectados por pseudomonas isolada ou pseudomonas associada ou não a outras bactérias gram-negativas. Os fungos filamentosos tiveram uma forte associação com trauma físico, uso de solução salina para a limpeza de lentes de contato teve uma forte associação com bactérias gram- negativas e trauma biológico, uma forte associação com a infecção combinada. Conclusão: Bactérias Gram negativas (em especial, Pseudomonas) seguido por fungos filamentosos (Fusarium sp) foram os agentes etiológicos mais freqüentemente encontrados em nosso estudo. O Cross-linking foi eficaz em interromper o melting de córnea. Pseudomonas aeruginosa foi o agente isolado mais frequente em usuários de lente de contato hidrofílica.

Microbiological and epidemiological study of infectious keratitis in children and adolescents / Estudo microbiológico e epidemiológico da ceratite infecciosa em crianças e adolescentes

ABSTRACT Purpose: To analyze epidemiological and microbiological aspects of microbial keratitis in children and adolescents. Methods: This retrospective cohort study was conducted at the Department of Ophthalmology and Visual Science, Escola Paulista de Medicina, Universidade Federal de São Paulo, between July 15, 1975, and December 31, 2010. We analyzed corneal samples from 859 patients with clinical suspicion of infectious keratitis, comparing epidemiological and microbiological characteristics of bacterial keratitis with those of non-bacterial and non-viral keratitis. We also compared Gram-positive and Gram-negative pathogens in patients with bacterial keratitis. We created a susceptibility profile of the bacterial microorganisms studied. Results: Of the 859 patients, 346 (40.3%) showed positive culture results for non-viral microorganisms. Teenagers (13-18 years) made up the group with the highest number of patients with keratitis (164, 47.4%). The most frequent risk factors for keratitis were trauma (33.5%) and previous ocular surgery (24.9%). Gram-positive bacteria (71.8%) were the most often isolated, with coagulase-negative Staphylococcus (23.8%) the most prevalent microorganism. Logistic regression analysis showed age (p=0.002), topical antimicrobial drug use (p=0.01), and trauma due to non-chemical burns (p=0.005) were risk factors for non-bacterial keratitis. Age (p=0.01) was also a risk factor for Gram-negative bacterial keratitis. Conclusion: Our study showed that in the age range studied, the prevalence of keratitis caused by Gram-negative bacteria or by the non-viral microorganisms evaluated increases with age. Previous use of topical antimicrobial drug and trauma due to non-chemical burns are associated with non-bacterial keratitis. Knowledge of the risk factors and the microorganisms involved may help improve treatment of keratitis in children and adolescents and minimize visual impairment.

RESUMO Objetivos: Descrever o perfil epidemiológico e microbiológico de ceratite microbiana em crianças e adolescentes. Métodos: Estudo retrospectivo tipo coorte, utilizando fichas laboratoriais de pacientes, atendidos no Departamento de Oftalmologia e Ciências Visuais - Escola Paulista de Medicina - Universidade Federal de São Paulo, entre 15 de julho de 1975 a 31 de dezembro de 2010. Foram comparados pacientes com ceratite bacteriana e não bacteriana (não viral). Entre os pacientes com ceratite bacteriana, foram comparados aqueles em que a ceratite foi causada por bactérias Gram positivas e Gram negativas. O perfil de sensibilidade dos microrganismos bacterianos aos antimicrobianos também foi estudado. Resultados: Foram analisadas amostras corneanas de 859 pacientes com suspeita clínica de ceratite infecciosa, na faixa etária estudada. Destes, 346 (40,3%) apresentaram resultados de culturas positivas para microrganismos não virais. Adolescentes (13 a 18 anos) compuseram o grupo com maior número de pacientes com ceratite (164-47,4%). Os principais fatores de risco foram trauma (33,5%) e cirurgias oculares prévias (24,9%). Bactérias Gram positivas foram isoladas com maior frequência (71,8%), sendo prevalente o patógeno Staphylococcus coagulase negativo (23,8%). De acordo com a análise de regressão logística, idade (p=0,002), uso tópico de drogas antimicrobianas (p=0,01) e trauma por queimadura não química (p=0,005) foram fatores predisponentes para ceratite não bacteriana. Idade (p=0,01) também foi fator de risco para ceratite causada por bactérias Gram negativas. Conclusões: Nosso estudo mostrou que quanto maior a idade, na faixa etária estudada, maior a probabilidade da ceratite ser causada por bactérias Gram negativas e/ou por outros microrganismos não virais avaliados. O uso tópico de drogas antimicrobianas prévias e trauma devido à queimadura não química predispõe à ceratite não bacteriana. O conhecimento dos fatores de risco e dos microrganismos envolvidos resultarão em tratamento específico da ceratite em crianças e adolescentes, com menores danos visuais.

Fonsecaea pedrosoi as a rare cause of acute conjunctival ulceration / Fonsecaea pedrosoi como agente raramente associado à úlcera conjuntival aguda

ABSTRACT Conjunctival ulceration accompanied with secretion and pain was observed in a 30-year-old male, 3 days after a perforating corneal trauma. Cultures of conjunctival ulcer samples grew Fonsecaea pedrosoi, a major causative agent of chromoblastomycosis that is typically transmitted during trauma. The conjunctival ulcer was successfully treated with amphotericin B, itraconazole, and fluconazole. This case report summarizes the diagnosis and treatment of a conjunctival ulcer due to F. pedrosoi, which is a rare complication of contaminated ocular trauma. To the best of our knowledge, this is the first reported case of F. pedrosoi causing acute conjunctival ulceration in the literature.

RESUMO O quadro clínico de uma úlcera conjuntival acompanhada de secreção e dor foi observado em homem de 30 anos de idade, 3 dias após um trauma perfurante da córnea. As culturas de uma amostra retirada da úlcera conjuntival foi positiva para Fonsecaea pedrosoi, uma cromoblastomicose, geralmente transmitido após traumatismos. O caso foi tratado com sucesso com a anfotericina B, itraconazol e fluconazol. Este relato de caso reporta o diagnóstico e tratamento de uma úlcera conjuntival causada por F. pedrosoi, que raramente é visto nos olhos expostos a traumatismos contaminados. Até onde sabemos, este é o primeiro caso relatado na literatura de F. pedrosoi causando úlcera conjuntival aguda.

Epidemiologia de ceratites fúngicas tratadas com ceratoplastia penetrante através de achados histopatológicos / Epidemiology of fungal keratitis treated with penetrating keratoplasty by means of histopathologic findings

RESUMO Objetivo: Estudar, através do exame histopatológico, a epidemiologia de ceratites fúngicas tratadas com ceratoplastia penetrante terapêutica, enfatizando a presença de cirurgia ocular prévia. Métodos: Inicialmente, o estudo foi observacional e transversal de botões corneanos provenientes de ceratoplastia penetrante no período de 2006-2015 enviados para exame histopatológico ao banco de olhos do Hospital Geral de Fortaleza. Os tecidos foram corados com Hematoxilina-eosina, PAS ou Grocott, e examinados com microscópio óptico. Foram selecionados casos com diagnóstico histopatológico de ceratite fúngica. Após a selecão, realizamos revisão de prontuários buscando idade e sexo do paciente, data(s) do(s) transplante(s) por ceratite fúngica, diagnóstico clínico précirúrgico, presença/tipo de cirurgias anteriores e/ou posteriores. Incluímos 62 botões corneanos de 55 pacientes. Resultados: A maioria dos pacientes era do sexo masculino. Apenas 7 (11,29%) casos tiveram recidiva da infecção tratada cirurgicamente. 10 (16,13%) casos possuíam cirurgia ocular prévia a ceratite fúngica tratada por transplante. Nenhum caso teve ceratite fúngica como diagnóstico clínico pré-cirúrgico. A principal forma de fungo no exame histopatológico foi forma leveduriforme isolada, seguida pela leveduriforme associada à filamentosa. O aspecto predominante da membrana de Descemet foi livre de fungos. Conclusão: Demonstramos o potencial curativo das ceratites fúngicas quando tratadas com ceratoplastia penetrante terapêutica e uma possível associação do fator cirurgia ocular prévia ao desenvolvimento dessas infecções. Características do exame histopatológico foram abordadas diferente de outros estudos que, em sua maioria, citam apenas o exame microbiológico. A dificuldade no diagnóstico clínico précirúrgico foi ressaltada, o que pode ter contribuído com a evolução dos casos estudados para tratamento cirúrgico.

ABSTRACT Objective: To study, by means of histopathological examination, the epidemiology of fungal keratitis treated with penetrating keratoplasty therapy, emphasizing the presence of previous ocular surgery. Methods: Initially, we made an observational and cross-sectional study of corneal buttons from penetrating keratoplasty in the 2006-2015 period sent for histopathological examination at the Hospital Geral de Fortaleza Eye Bank.Tissues were stained with hematoxylin-eosin, PAS or Grocott, and examined with an optical microscope. We selected the cases with histopathological diagnosis of fungal keratitis. After the selection, we carried out a review of records seeking for age and sex of patients, date(s) of the graft(s) made to treat fungal keratitis, preoperative clinical diagnosis, presence/type of earlier and/or subsequent surgeries.We included 62 corneal buttons from 55 patients. Results: Most patients were male. Only 7 (11.29%) cases had recurrence of the surgically treated infection. 10 (16.13%) cases had eye surgery prior to fungal keratitis treated by transplant. No cases had fungal keratitis as preoperative clinical diagnosis.The main form of fungus in histopathological examination was isolated yeast form, followed by the yeast form associated with the filamentous form.The predominant aspect of Descemet's membrane was free of fungus. Conclusion: We demonstrated the healing potential of fungal keratitis when treated with penetrating therapeutic keratoplasty and the possible association of previous eye surgery factor for the development of these infections. Characteristics of histopathological examination have been approached differently from other studies that mostly specify only the microbiological examination. The difficulty in preoperative clinical diagnosis was highlighted, which may have contributed to the evolution of the cases studied for surgical treatment.

Correlação clínico-patológica em ceratites fúngicas e uma possível evidência de infecção oportunista: relato de dois casos / Clinicopathological correlation in fungal keratitis and a possible evidence of opportunistic infection: a report of two cases

RESUMO Os autores relatam dois casos de ceratite fúngica, que apresentam em comum no exame histopatológico: a ausência de epitélio, sem ulceração e a integridade da camada de Bowman com a presença de formas fúngicas no seu interior. São sugeridos prováveis mecanismos de recidiva das ceratites fúngicas, ressaltando uma possível "via externa não usual", por meio da penetração do fungo através da camada de Bowman íntegra. A existência desta infecção oportunista não está relatada na literatura oftalmológica e são necessárias mais evidências para que seja considerada como tal.

ABSTRACT The authors describe two cases of fungal keratitis which, upon histopathological examination, are found to have in common the absence of epithelium, without ulceration and the lack of integrity of the Bowman's layer with the presence of fungal forms in their interior. Through them the authors suggest probable mechanisms of recurrence of fungal keratitis, highlighting the possible existence of an "unusual external route" that would occur by the fungus penetration through the full Bowman layer. Although these findings appear not yet reported or recovered in the ophthalmic literature, the authors suggest that they could be possible opportunistic infection signals which, however, require more evidence to be considered as such.

Early post-LASIK flap amputation in the treatment of aggressive, branching keratitis: a case report / Amputação precoce de pedículo de LASIK para o tratamento de ceratite ramificada agressiva: relato de caso

ABSTRACT Infectious keratitis is rare following laser vision correction. We present a case of aggressive fungal keratitis caused by Aspergillus flavus, following laser in situ keratomileusis (LASIK) in the setting of a unique environmental risk factor. We describe the key features of the acute case presentation, which guided empirical medical and surgical treatment, resulting in the most favorable outcome found in the literature, to date.

RESUMO Ceratites infecciosas são raras após a correção da visão a laser. Apresentamos um relato de caso de uma ceratite fúngica agressivo por Aspergillus flavus, após ceratomileuse a laser "in situ" (LASIK) em situação peculiar de fator de risco ambiental. Nós descrevemos as principais características da apresentação caso agudo, que orientou tratamento médico e cirúrgico empírico, demonstrando resultado mais favorável do que o encontrado na literatura até o momento.

Sensibilidad antibiótica de bacterias obtenidas de queratitis e infecciones intraoculares en la Fundación Oftalmológica de Santander (FOSCAL), Floridablanca, Colombia / Antibiotic susceptibility patterns of bacteria isolated from keratitis and intraocular infections at Fundación Oftalmológica de Santander (FOSCAL), Floridablanca, Colombia

Introducción. La resistencia bacteriana es crítica para la selección de los antibióticos en el tratamiento de las infecciones, por ello es vital conocer su estado actual en nuestro medio. Objetivo. Determinar la sensibilidad antibiótica bacteriana in vitro obtenida de los cultivos de queratitis e infecciones intraoculares. Materiales y métodos. Se llevó a cabo un estudio retrospectivo en la Fundación Oftalmológica de Santander (FOSCAL), entre junio de 2011 y enero de 2012. Resultados. Se examinaron 92 muestras. Se identificaron 110 bacterias, 27 hongos y 12 amebas de vida libre. Del total de bacterias Gram positivas, 1,1 %, 0 %, 1,1 %, 16,9 %, 29,3 % y 85 % fue resistente a imipenem, moxifloxacina, gatifloxacina, levofloxacina, ciprofloxacina y tobramicina, respectivamente, mientras que la resistencia a estos mismos fármacos se presentó, respectivamente, en 0 %, 8,3 %, 0 %, 0 %, 18,2 % y 27,3 % de las bacterias Gram negativas. Los porcentajes de resistencia de los estafilococos positivos para coagulasa resistentes a la meticilina fueron 0 %, 0 %, 0 %, 7 %, 17 % y 100 %, respectivamente, y los porcentajes de los estafilococos negativos para coagulasa resistentes a la meticilina fueron 3 %, 0 %, 0 %, 24 %, 44 % y 100 %, respectivamente. Los porcentajes de resistencia bacteriana globales (tanto para bacterias Gram positivas como para Gram negativas) a imipenem, moxifloxacina, gatifloxacina, levofloxacina, ciprofloxacina y tobramicina fueron 1 %, 1 %, 1 %, 15,1 %, 28 % y 64,5 %, respectivamente. Conclusiones. Los niveles de resistencia bacteriana para imipenem, moxifloxacina y gatifloxacina fueron menores que para levofloxacina, ciprofloxacina y tobramicina. Los niveles de resistencia para la tobramicina fueron muy altos, lo que pone en duda su utilidad clínica en las infecciones oculares en nuestro medio.

Introduction: Bacterial resistance is critical for the selection of antibiotics in the treatment of infections, so it is vital to know its current status in our geographical area. Objective: To determine in vitro antibiotic susceptibility of bacterial isolates obtained from keratitis and intraocular infections. Materials and methods: A retrospective study of microbiological tests in Fundación Oftalmológica de Santander (FOSCAL) was carried out between June, 2011, and January, 2012. Results: A total of 92 samples were examined and 110 bacteria, 27 fungi and 12 free-living amoebae were identified. Polymicrobial infections constituted 50% of the total; 1.1%, 0%, 1.1%, 16.9%, 29.3% and 85% of Gram-positive bacteria were resistant to imipenem, moxifloxacin, gatifloxacin, levofloxacin, ciprofloxacin and tobramycin, respectively, while 0%, 8.3%, 0%, 0%, 18.2% and 27.3% of Gram-negative bacteria were resistant to imipenem, moxifloxacin, gatifloxacin, levofloxacin, ciprofloxacin and tobramycin, respectively. For methicillin-resistant coagulase-positive staphylococci, resistance percentages to imipenem, moxifloxacin, gatifloxacin, levofloxacin, ciprofloxacin and tobramycin were 0%, 0%, 0%, 7%, 17% and 100%, respectively. For methicillin-resistant coagulase-negative staphylococci, resistance percentages to imipenem, moxifloxacin, gatifloxacin, levofloxacin, ciprofloxacin and tobramycin were 3%, 0%, 0%, 24%, 44% and 100%, respectively. Overall bacterial resistance to imipenem, moxifloxacin, gatifloxacin, levofloxacin, ciprofloxacin and tobramycin, for both Gram-positive and Gram-negative, was 1%, 1%, 1%, 15.1%, 28% and 64.5%, respectively. Conclusions: The levels of bacterial resistance to imipenem, moxifloxacin and gatifloxacin were lower than for levofloxacin, ciprofloxacin and tobramycin. The levels of resistance to tobramycin were very high, which calls into question its usefulness in this region of our country.

Queratitis micótica: identificación de especies en Valparaíso / Fungal keratitis: identification of species in Valparaíso

Introducción: La queratitits micótica puede ser provocada por una amplia variedad de especies, tanto por hongos filamentosos como levaduras. Objetivo: Conocer las especies involucradas en queratitits micótica durante el decenio 2003-2013 en la ciudad de Valparaíso. Materiales y método: Se revisaron los casos de queratitis micótica diagnosticados durante el periodo 2003-2013 en la cátedra de Micología de la Universidad de Valparaíso. Las muestras fueron tomadas con tórulas estériles, las cuales fueron sembradas en agar sabouraud y una vez obtenido el desarrollo de las especies se traspasaron a medios de cultivo especiales. La identificación se realizó por morfofisiología y en el caso de Candida además por pruebas bioquímicas. Resultados: Se diagnosticaron 10 pacientes con queratitis micótica, de ellos 6 fueron hombres. 8 pacientes tenían el antecedente de lesión traumática corneal. Las especies más frecuentes fueron Aspergillus fumigatus y Fusarium oxysporum. También se aislaron especies como Fusarium dimerumy Exophiala jeanselmei. Conclusión: De los 10 casos de queratitis micótica diagnosticados, la mayoría tenía el antecedente de lesión traumática corneal. Los géneros más aislados fueron Aspergillus y Fusarium.

Introduction: Fungal keratitis can be caused by a wide variety of species, either filamentous as yeast fungi. Objective: To determine the species involved in fungal keratitis during the decade 2003-2013 in the city of Valparaiso. Materials and Methods: The cases of fungal keratitis diagnosed in the depar tment of Mycology, University of Valparaiso, were reviewed for the period 2003-2013. Samples were taken with sterile swabs, which were seeded in Sabouraud agar and after obtaining the development of species, were transferred to special culture media. Identification was performed by morphophysiology and in the case of Candida further biochemical tests. Results: 10 patients were diagnosed with fungal keratitis, of which 6 were men. 8 patients had a history of traumatic corneal injury. The most frequent species were Aspergillus fumigatus and Fusarium oxysporum. Species Fusarium dimerum and Exophiala jeanselmei were also isolated. Conclusion: Of the 10 cases of fungal keratitis diagnosed, most had a history of traumatic corneal injury. The most isolated genus were Aspergillus and Fusarium.

Self-inflicted Chronic Bacterial Keratoconjunctivitis Using Self Semen

This case report describes a case of self-inflicted chronic bacterial keratoconjunctivitis involving the patient's own semen. A 20-year-old male soldier was referred to our clinic for the evaluation of refractory chronic bacterial conjunctivitis. Over the previous 4 months, he had been treated for copious mucous discharge, conjunctival injection, and superficial punctate keratitis in both eyes at an army hospital and a local eye clinic. Despite the use of topical and systemic antibiotics according to the results of conjunctival swab culture, there was no improvement. During the repeated smear and culture of conjunctival swabs, surprisingly, a few sperm were detected on Gram staining, revealing that the condition was self-inflicted bacterial keratoconjunctivitis involving the patient's own semen. Thus, in cases of chronic keratoconjunctivitis that do not respond to appropriate antibiotic treatment, self-inflicted disease or malingering should be considered.

Lab diagnosis and evaluation of fungal keratitis

This study is aimed to evaluate the usefulness of Sabouraud dextrose agar in the confirmatory diagnosis of suspected cases of fungal keratitis. Pre-designed prospective study of corneal scraping obtained after detailed slit-lamp examination and documentation from all patients seen for non-viral microbial infective keratitis. This study was conducted in the Department of Ophthalmology, JPMC, Karachi from July 2008 to June 2011. 128 cases of non-viral microbial infective keratitis. Corneal scraping of 128 patients with microbial keratitis Smears of corneal scraping were stained with Gram's Method and inoculated specimens on Sabouraud dextrose agar [SDA] and incubated for 3-4 days. In a series of suspected cases of fungal keratitis, 119/128 [75% patients] had positive results for fungus in corneal scrapings by direct microscopy using Grams staining method and culture on Sabouraud dextrose agar [SDA]. 43% males and 32% females had Candidial keratitis and 11% males and 7% females had Fusarium infection. Other samples showed presence of Gram positive cocci on smears and were negative for SDA. Fungal keratitis continue to be an important cause of ocular morbidity, since it becomes difficult to clinically diagnose and differentiate between bacterial and mycotic keratitis in complicated cases. It is better to use a standard culture medium like Sabouraud agar [SDA] when confirming ocular mycosis

Molecular characterization of drug-resistant and drug-sensitive Aspergillus isolates causing infectious keratitis.

Purpose: To study the susceptibilities of Aspergillus species against amphotericin B in infectious keratitis and to find out if drug resistance had any association with the molecular characteristics of the fungi. Materials and Methods: One hundred and sixty Aspergillus isolates from the corneal scrapings of patients with keratitis were tested for susceptibilities to amphotericin B by broth microdilution method. These included Aspergillus flavus (64 isolates), A. fumigatus (43) and A. niger (53). Fungal DNA was extracted by glass bead vertexing technique. Polymerase chain reaction (PCR) assay was standardized and used to amplify the 28S rRNA gene. Single-stranded conformational polymorphism (SSCP) of the PCR product was performed by the standard protocol. Results: Of the 160 isolates, 84 (52.5%) showed low minimum inhibitory concentration (MIC) values (≤ 1.56 μg/ml) and were designated as amphotercin B-sensitive. Similarly, 76 (47.5%) had high MICs (≥ 3.12 μg/ml) and were categorized as amphotericin B-resistant. MIC50 and MIC90 values ranged between 3.12-6.25 μg/ml and 3.12-12.5 μg/ml respectively. A. flavus and A. niger showed higher MIC50 and MIC90 values than A. fumigatus. The SSCP pattern exhibited three extra bands (150 bp, 200 bp and 250 bp each) in addition to the 260 bp amplicon. Strains (lanes 1 and 7) lacking the 150 bp band showed low MIC values (≤ 1.56 μg/ml). Conclusion: A. niger and A. flavus isolates had higher MICs compared to A. fumigatus, suggesting a high index of suspicion for amphotericin B resistance. PCR-SSCP was a good molecular tool to characterize Aspergillus phenotypes in fungal keratitis.

Mycotic corneal ulcers in upper Assam.

Purpose: To study the association of various risk factors and epidemiological variables of mycotic keratitis treated at a tertiary referral hospital of upper Assam. Materials and Methods: In this hospital-based prospective study a total of 310 consecutive corneal ulcer cases attending the ophthalmology outpatient department of Assam Medical College were enrolled between April 2007 and March 2009. After clinical and slit-lamp biomicroscopic examination in all suspected cases, smears and culture examination for fungus was done to establish the etiology. Demographic information and associated probable risk factors of individual cases were noted in a predesigned questionnaire. Results: In 188 (60.6%) cases fungal etiology could be established. Out of them 67.6% were males. The most commonly affected age group was 41-50 years (25.5%). The maximum (23.4%) cases were reported during the paddy harvesting season in Assam (January and February). Fungal element could be demonstrated in 65.2% cases in direct potassium hydroxide (KOH) mount. The commonest predisposing factor was corneal injury (74.5%). While diabetes was a significant systemic predisposing factor in mixed bacterial and fungal infections in 11.1% cases, blocked naso-lacrimal duct was the local predisposing factor in 11.1% of cases. Fusarium solani (25%) was the commonest isolate followed by Aspergillus species (19%), Curvularia species (18.5%) and Penicillium species (15.2%). Yeasts were isolated in 2.7% (n=5) cases. Conclusions: Ocular trauma was the commonest cause of fungal corneal ulcer in Assam and Fusarium solani was the commonest species responsible for it. Most of the mycotic ulcer cases come from rural areas including the tea gardens.

A Case of Stenotrophomonas maltophilia Keratitis Effectively Treated with Moxifloxacin

A 70-year-old man with a long history of diabetes mellitus presented to our hospital (Department of Ophthalmology, Sahm Yook Medical Center, Seoul, Korea) complaining of severe ocular pain and visual disturbance in his left eye that had started three days prior to admission. A round 3.7 x 5.0 mm dense central stromal infiltrate with an overlying epithelial defect was noted on slit-lamp examination. Following corneal scrapings and culture, topical 0.5% moxifloxacin and 0.5% tobramycin were administered hourly. A few days later, Stenotrophomonas maltophilia was isolated in a bacterial culture from a corneal specimen. According to the results of susceptibility tests, topical 0.5% moxifloxacin was given every hour and 0.5% tobramycin was stopped. The patient's clinical features improved steadily with treatment. The corneal epithelium healed rapidly, and the infiltrate resolved within four weeks of the initiation of treatment. The patient's best corrected visual acuity improved from hand motion to 20 / 25.

A Case of Stenotrophomonas maltophilia Keratitis Effectively Treated with Moxifloxacin

A 70-year-old man with a long history of diabetes mellitus presented to our hospital (Department of Ophthalmology, Sahm Yook Medical Center, Seoul, Korea) complaining of severe ocular pain and visual disturbance in his left eye that had started three days prior to admission. A round 3.7 x 5.0 mm dense central stromal infiltrate with an overlying epithelial defect was noted on slit-lamp examination. Following corneal scrapings and culture, topical 0.5% moxifloxacin and 0.5% tobramycin were administered hourly. A few days later, Stenotrophomonas maltophilia was isolated in a bacterial culture from a corneal specimen. According to the results of susceptibility tests, topical 0.5% moxifloxacin was given every hour and 0.5% tobramycin was stopped. The patient's clinical features improved steadily with treatment. The corneal epithelium healed rapidly, and the infiltrate resolved within four weeks of the initiation of treatment. The patient's best corrected visual acuity improved from hand motion to 20 / 25.

Is inclusion of Sabouraud dextrose agar essential for the laboratory diagnosis of fungal keratitis.

Purpose: To determine whether the inclusion of Sabouraud dextrose agar (SDA) is essential in the diagnosis of fungal keratitis. Materials and Methods: Corneal scrapings of 141 patients with microbial keratitis were smeared and cultured. Sheep blood agar (BA), chocolate agar (CA), SDA, non-nutrient agar (NNA) with Escherichia coli overlay, and brain heart infusion broth (BHI) were evaluated for time taken for growth and cost. The media were also evaluated experimentally for rate of growth and time taken for identification. Results: Twenty-six of 39 patients positive for fungus in corneal scrapings by microscopy were culture-positive. Fungus grew on BA in 22/39, on CA in 18/39, on SDA in 17/39, on NNA in 17/39, and on BHI in 13/39 cases. Growth on SDA was higher in ulcers with larger infiltrate (6/18 versus 9/13, P = 0.04). Estimated saving with inclusion of only BA/CA was Rs. 600 per patient. Performance of all media was similar in in vitro experiment although the characteristic spores and color were seen earlier on SDA. Conclusion: Fungal keratitis can be reliably confirmed on BA or CA, which support growth of both bacteria and fungus.

Two Cases of Corneal Ulcer due to Methicillin-Resistant Staphylococcus aureus in High Risk Groups

Considering the popular use of antibiotic-containing eyedrops in Korea, it is important to know the emerging antibiotic-resistant strains of bacteria before treating infectious eye diseases. This is especially important in high-risk groups because of the high incidence of resistant infections and the subsequent treatment requirements. We report two cases of methicillin-resistant Staphylococcus aureus (MRSA) corneal ulcers in high-risk groups. The first case involved a patient who had keratitis after using antibiotic- and steroid-containing eyedrops to treat a corneal opacity that developed after repeated penetrating keratoplasty. The second case involved a patient who used antibiotic-containing eyedrops and a topical lubricant on a regular basis for >1 month to treat exposure keratitis due to lagophthalmos. The second patient's problems, which included a persistent superficial infiltration, developed after brain tumor surgery. Both cases showed MRSA on corneal culture, and the corneal ulcers improved in both patients after the application of vancomycin-containing eyedrops. In conclusion, MRSA infection should be considered in corneal ulcers that have a round shape, mild superficial infiltration, and slow progression, especially in high-risk groups. This report includes descriptions of the characteristic features, antibiotic sensitivities, prevention, and successful treatment with vancomycin-containing eyedrops for MRSA corneal ulcers.

Bacteriological studies on pathogenic microorganisms of blood, sputum, wound, and corneal samples from Indian patients

The present work aim to study the various pathogenic microorganisms from blood, wound, corneal and sputum samples from patients hosted in Tiruchirappalli area, Tamilnadu, India, during the month of February 2007 to June 2007. A total of 1031 samples were collected from patients. They include 195 blood samples, 310 sputum samples, 414 pus samples from wounds and 112 corneal swabs. The samples were inoculated into different culture media. Various bacterial and fungal colonies were isolated and identified by relevant staining methods and various biochemical utilization tests and also their antibiotics sensitivity were analyzed. Based on their antibiotic sensitivity pattern, bacterial and fungal pathogens were identified as sensitive and resistant strains. Mostly single pathogen was isolated from all the samples. Gram positive bacteria were found as common causative agent than gram negative bacteria. S. aureus and Klebsiella spp., were frequently isolated from all the samples. In blood samples, Klebsiella spp., E. coli, and Salmonella species were predominantly isolated, p-haemolytic Streptococci and S. aureus were predominantly isolated from sputum samples and Mycobacterium tuberculosis was isolated only in few samples. In wound samples S. aureus, Proteus mirabilis and P. vulgaris were predominantly isolated than E. coli, Enterobacter spp., P. aeruginosa, Serratia marcescens and Citrobacter divergens. Pseudomonas spp., Fusarium spp., Aspergillus Niger and A.flavus isolated from the corneal swabs. Antibiotic sensitivity tests were analyzed in those bacteria and fungal strains. The bacterial strains particularly S. aureus and Klebsiella spp., were highly resistant to 9 antibiotics where as E. coli, Enterobacter spp., and Streptococcus spp., were highly resistant to 10 antibiotics. The widespread uses of antibiotics are strongly associated with the development of bacterial drug resistance and also emerging of multidrug resistant strains

Topical and Oral Voriconazole in the Treatment of Fungal Keratitis

We describe two patients with fungal keratitis refractory to standard antifungal therapy whose conditions were managed with voriconazole. The first case is a patient with endophthalmitis and corneal ulcer due to Candida parapsilosis after receiving a corneal transplant. The patient was treated with amphotericin but showed no signs of improvement. Topical voriconazole, oral voriconazole, and intravitreal voriconazole yielded signs of improvement. The second case is a 63-year-old male who underwent a month of empiric treatment with 0.2% topical amphotericin for fungal keratitis but showed no signs of improvement. Treatment was then provided with 1% voriconazole. Both cases showed effective treatment with voriconazole. Voriconazole may be considered as a new method to treat fungal keratitis refractory to standard antifungal therapy.

Contaminação de halos doadores córneo-esclerais em ceratoplastia penetrante no Hospital de Clínicas de Porto Alegre / Positive corneoscleral rim culture in penetrating keratoplasty at the Porto Alegre Clinical Hospital

OBJETIVOS: Avaliar a incidência de positividade de culturas de halos doadores córneo-esclerais preservados em Optisol GS, identificar os patógenos envolvidos, a sensibilidade dos mesmos à gentamicina e a ocorrência de infecções em olhos receptores. MÉTODOS: Foram analisadas retrospectivamente 163 culturas de halos córneo-esclerais cujos botões corneanos foram utilizados em transplantes de córnea no Hospital de Clínicas de Porto Alegre entre janeiro de 2001 e janeiro de 2003. Os halos foram divididos em dois segmentos, metade inoculada em meio Sabouroud e a outra metade em tioglicolato, com posterior semeadura em ágar-sangue, ágar-chocoloate e meio de MacConkey, conforme necessidade para identificação dos patógenos. Os prontuários dos pacientes receptores foram revisados. RESULTADOS: Dos 163 halos analisados, 11 apresentaram culturas positivas, correspondendo a 6,7 por cento do total. Destes, quatro foram por Staphylococcus epidermidis, um por Staphylococcus aureus, um por Serratia sp, um por Pseudomonas aeruginosa e os outros quatro por diferentes subtipos de Candida (dois por Candida sp, um por Candida albicans e um por Candida parapapilosis). No antibiograma, todas as bactérias apresentaram-se resistentes à gentamicina. Nenhum olho que recebeu córnea com cultura positiva apresentou infecção após a cirurgia. CONCLUSÕES: Baixos índices de positividade de cultura de halos utilizados em transplantes de córnea no Hospital de Clínicas de Porto Alegre foram encontrados. Os patógenos mais freqüentemente identificados não apresentam boa cobertura pelos antimicrobianos presentes nos meios de preservação. A cultura de halos corneanos é recomendada para auxílio no tratamento de possível infecção ocular pós-cirúrgica.

PURPOSE: To determine the incidence of positive corneoscleral rim cultures preserved in Optisol GS medium, to identify pathogens involved and possible recipient eye infection. METHODS: A hundred sixty-three corneoscleral rim cultures penetrating keratoplasties performed from January 2001 to January 2003 in the Hospital de Clínicas de Porto Alegre were reviewed. Enucleations and corneal storage were done as aseptic as possible and gentamicin 0.3 percent was instilled. Corneoscleral rim was divided into two segments, half was inoculated into Sabouraud broth and the other half into thioglycolate broth; inoculation into blood agar, chocolate agar and MacConkey agar was done later if necessary for pathogen identification. The receiver's eye data were reviewed. RESULTS: There were eleven positive cultures (6.7 percent) out of 163 evaluated corneoscleral rim cultures. Of these, four were Staphylococcus epidermidis, one was Staphylococcus aureus, one was Serratia sp., one was Pseudomonas aeruginosa and the other four were different subtypes of Candida (two Candida sp., one Candida albicans and one Candida parapapilosis). All pathogens were resistant to gentamicin. None of the eleven cases of positive corneoscleral rim cultures resulted in ocular infection at the receiver's eyes (six months follow-up). CONCLUSIONS: We found low rates of positive corneoscleral rim cultures after penetrating keratoplasty at the Porto Alegre Clinical Hospital. The most frequent involved pathogens were Staphylococcus sp and Candida sp. Although we did not identify any postoperative infection at the receiver's eyes, we recommend corneoscleral rim culture for guidance of postoperative infection, a rare but possible devastating ocular event.