Background: Fungal keratitis is a serious ocular infection that can cause corneal scarring and blindness. Currently, diagnosis of fungal pathogens remains a difficult problem. Objectives: To investigate the application of semi-nested PCR targeted ITS genes for detection of fungal agents causing keratitis. Material and method: Ten identified fungal strains, 4 bacterial strains, 20 scraping samples from patients with suspected fungal keratitis and 2 scraping samples from patients with suspected bacterial keratitis were tested using semi-nested PCR. Results: Semi-nested PCR showed positive results for the samples of identified fungal strains and for the 20 scraping samples from patients with suspected fungal keratitis. Neither samples of bacterial strains nor scraping samples from suspected bacterial keratitis patients gave positive PCR results. Conclusion: Semi-nested PCR is a robust tool for specific and rapid detection of fungal agents causing keratitis.
Fungal keratitis ; semi-nested PCR

Fungal corneal ulcer could lead to a devastating outcome. The relative scarcity of readily available, inexpensive but effective topical antifungal drug has left many ophthalmologists desperate and frustated in treating the condition. The use of oral itraconazole has long been proven to be of clinical value in a number of forms of mycoses. Its safety profile is extremely good with minimal reported adverse effects. We investigated an aqueous form of itraconazole which we prepared into a 0.1 mg/ml concnetration and compared its efficacy as a topical antifungal against the standard drug. Natamycin 5 percent in the treatment of fungal keratitis in rabbits. A randomized animal trial was done using 24 rabbit eyes divided into 2 treatment groups. All rabbit corneas were inoculated with Aspergillus flavus and treated after 48 hours with either Topical itraconazole or Natamycin 5 percent for a period of 2 weeks. Results showed inhibition of the disease with both treatment groups. We also found no significant difference between the severity, progression and remission of the keratitis between both treatment groups clinically and statistically. Topical itraconazole 0.1 mg/ml was shown to be comparable to our standard topical antifungal Natamycin 5 percent in treating fungal keratitis. (Author)
Animal ; ASPERGILLUS KERATITIS ; OPHTHALMOLOGY ; ASPERGILLUS FLAVUS ; NATAMYCIN ; ITRACONAZOLE ; FUNGAL CORNEAL ULCER ; FUNGAL KERATITIS

PURPOSE: To measure the secretion of IL-8 from cultured human keratocytes after inoculation of conidia of A. fumigatus, and to compare the change of secretion of IL-8 following application of amphotericin B and dexamethasone. METHODS: Human corneal keraoctytes were cultured independently in vitro. The specimens were divided into 4 groups : Group I with only pure culture as control, Group II with conidia of A. fumigatus, Group III with conidia of A. fumigatus and amphotericin B, and Group IV with conidia of A. fumigatus and dexamethasone. The supernatants were aspirated from each group at different time intervals, and then were assayed for IL-8. RESULTS: Group II showed increased secretion of IL-8, at all selected time intervals except 12-h, in comparison with other three groups. Group III secreted IL-8 significantly less than the other groups(p<0.01). Group IV secreted IL-8 less than Group I and II at all selected time points(p<0.01), but more than Group III at the other time points except for 72-h(p<0.01). CONCLUSIONS: The secretion of IL-8 increased in the early stage of fungal keratitis but decreased in case of the administration of amphotericin B or dexamethasone. Amphotericin B was more potent than dexamethasone in decreasing the secretion of IL-8.
Amphotericin B* ; Aspergillus fumigatus* ; Aspergillus* ; Dexamethasone* ; Humans* ; Interleukin-8* ; Keratitis ; Spores, Fungal

This study was performed for the standardization and proper selection of effective antifungal agents by measuring the minimal inhibitory concentra-tions[MICs]of antifungals to fungi, separated from keratitis patients.Two strains of A.fumigatus and single strain of F.solani, A.falciforme, and A . alternata were used for this test.Standard powders of miconazole, itraconazole, clotrimazole, ketoconazole, and amphotericin B were used as antifungal agents. Microscopic and macroscopic measurements of MIC after 24, 48 and 72 hours of inoculation[105 conidia /ml]into YNB broth with culture temperature of 25 degrees C were performed by use of broth microdilution method.The results are as follows : itraconazole, amphotericin B, and clotrimazole were effective against A.fumigatus.F.solani showed resistance to all kinds of antifungal agents.A.falciforme and A.alternata were sensitive to amphotericin B and itraconazole, respectively. Further studies may be needed for the standardized measurement of MIC against filamentous fungi and also for in Vitro-in Vivo correlations for the treatment of fungal keratitis.
Amphotericin B ; Antifungal Agents* ; Clotrimazole ; Fungi ; Itraconazole ; Keratitis* ; Ketoconazole ; Miconazole ; Powders ; Spores, Fungal

Adult ; Antifungal Agents ; therapeutic use ; Eye Infections, Fungal ; diagnosis ; drug therapy ; Humans ; Keratitis ; diagnosis ; drug therapy ; Male

PURPOSE: To describe the successful treatment of a series of fungal ocular infections with voriconazole. METHODS: A retrospective chart review was conducted for six eyes of five patients: four eyes of four patients with fungal keratitis, and two eyes of one patient with fungal endophthalmitis who were treated with topical and intracameral voriconazole. A battery of analyses on corneal isolates, clinical outcomes, duration of treatment, time to remission, complications, concomitant medication, and visual acuity at the initial and last visits was performed. RESULTS: Six eyes of five patients were treated in total. Three eyes with fungal keratitis were treated with 5% topical voriconazole, and infection was controlled. Therapeutic penetrating keratoplasty was performed on one eye with fungal keratitis due to impending corneal perforation; this eye was treated successfully with topical voriconazole without recurrent inflammation. Both eyes of one patient with fungal endophthalmitis were given an intracameral voriconazole (100 microgram/0.1 cc) injection twice in the right eye and once in the left eye, and infection was controlled with continued topical voriconazole therapy. There was no clinically remarkable corneal epithelial toxicity from topical application. No corneal endothelial toxicity was observed after an intracameral injection of voriconazole. CONCLUSIONS: Voriconazole is known to penetrate efficiently into ocular tissues. As shown in our case series, voriconazole is a new, promising therapy for intractable fungal ocular infection.
Endophthalmitis ; Eye ; Eye Infections, Fungal ; Humans ; Inflammation ; Keratitis ; Keratoplasty, Penetrating ; Pyrimidines ; Retrospective Studies ; Time-to-Treatment ; Triazoles ; Visual Acuity

Colletotrichum is mainly a fungal pathogen of plants, but sporadic cases of human infection have been reported recently. Most of them are fungal keratitis and only a few cases have been reported worldwide. A 63-year-old female farmer developed foreign body sensation and watering in her left eye following trauma by rice leaves. At presentation, her visual acuity decreased and corneal ulcer and inflammation in anterior chamber were observed on a slit lamp examination. Numerous hyphae were found on Gram stain and a rapidly growing mold with cup-shaped acervuli and falcate and nonseptate conidia was observed on fungal culture. As morphological findings did not lead to definite differentiation of the organism, sequencing of the D1-D2 domain of 28S rDNA was performed. It proved to be Colletotrichum species and the patient was treated with amphotericin and natamycin eye drop, but complicated by acute glaucoma. This is the first report of Colletotrichum keratitis in Korea and suggests that its infection should be considered in patients with fungal keratitis.
Amphotericin B ; Anterior Chamber ; Colletotrichum* ; Corneal Ulcer ; DNA, Ribosomal ; Female ; Foreign Bodies ; Fungi ; Glaucoma ; Humans ; Hyphae ; Inflammation ; Keratitis* ; Korea ; Middle Aged ; Natamycin ; Sensation ; Spores, Fungal ; Visual Acuity ; Water

INTRODUCTIONCorneal infection is the most common cause of profound ocular morbidity leading to blindness worldwide. Corneal infection in children is difficult to diagnose and treat, as they are unwilling and sometimes unable to cooperate during active management. This study analyses the prevalence, microbiology, demography, therapeutic and visual outcome of infectious microbial keratitis in the paediatric age group seen at a tertiary eye care hospital in south India.

MATERIALS AND METHODSA retrospective review of all cases presenting with keratitis to the ocular microbiology and cornea service at Aravind Eye Hospital, Coimbatore, from February 1997 to January 2004, was done to screen the patients for microbial keratitis. Their records were further analysed for clinical and microbiological details. Cases with culture-proven non-viral keratitis in children

RESULTSOf the 310 patients who attended the cornea clinic, 97 (31.2%) patients were confirmed to be positive for microbial keratitis. 54.6% of cases were male. The most common predisposing cause of ulceration was trauma (69%) with organic matter. Pure bacterial cultures were obtained from 64 (65.9%) eyes, whereas pure fungal cultures were obtained from 37 (38.1%) eyes. Four (4.1%) eyes showed mixed growth.

CONCLUSIONThe most commonly isolated organism was Pseudomonas aeruginosa. The most common predisposing cause of infectious microbial keratitis was corneal trauma. Early stage of diagnosis and formulation of an uncompromising management protocol can prevent profound visual morbidity.

Adolescent ; Child ; Child, Preschool ; Eye Infections, Bacterial ; diagnosis ; therapy ; transmission ; Eye Infections, Fungal ; diagnosis ; therapy ; transmission ; Female ; Humans ; India ; Infant ; Keratitis ; diagnosis ; etiology ; microbiology ; therapy ; Male

Purpureocillium(P) lilacinum is a ubiquitous, saprophytic filamentous fungus that is infrequently reported in keratitis and cutaneous infections. However, the microbiological characterization of the culture isolates is limited in Korea. A 56-year-old male who suffered a pine needlestick to his right eye 10 days previously presented with ocular opacity and pain. A microscopic examination of a corneal scraping by Gram staining and calcofluor white staining was negative for bacteria and fungi. Fungal culture yielded pure white cottony molds on Sabouraud's dextrose agar after a 3-day incubation. Microscopic examination further revealed a mixture of a verticillate arrangement of phialides resembling the Penicillium structure and sparsely branched conidiophores bearing single to small clusters of conidia. This was initially presumed to be a species of Penicillium but the colonies never turned green with further incubation. It was subsequently identified as P. lilacinum by 28S rDNA sequencing and MALDI-TOF mass spectrometry. Antifungal susceptibility test revealed that this organism was resistant to flucytosine, amphotericin B, fluconazole, itraconazole, voriconazole, and caspofungin. After treatment with topical 5% voriconazole and oral itrazonazole combined with multiple debridements for 2 weeks, the patient was discharged with improved visual acuity. We thus report the first case of P. lilacinum infection that required molecular identification due to mixed conidiogenesis features and that showed laboratory-confirmed antifungal resistance in Korea.
Agar ; Amphotericin B ; Bacteria ; Debridement ; DNA, Ribosomal ; Drug Resistance ; Fluconazole ; Flucytosine ; Fungi ; Glucose ; Humans ; Itraconazole ; Keratitis* ; Korea ; Male ; Mass Spectrometry ; Middle Aged ; Needlestick Injuries ; Penicillium ; Sequence Analysis, DNA ; Spores, Fungal ; Visual Acuity ; Voriconazole

Phialemonium keratitis is a very rare case and we encountered a case of keratitis caused by Phialemonium obovatum (P. obovatum) after penetrating injury to the cornea. This is the first case report in the existing literature. A 54-year-old male was referred to us after a penetration injury, and prompt primary closure was performed. Two weeks after surgery, an epithelial defect and stromal melting were observed near the laceration site. P. obovatum was identified, and then identified again on repeated cultures. Subsequently, Natacin was administered every two hours. Amniotic membrane transplantation was performed due to a persistent epithelial defect and impending corneal perforation. Three weeks after amniotic membrane transplantation, the epithelial defect had completely healed, but the cornea had turned opaque. Six months after amniotic membrane transplantation, visual acuity was light perception only, and corneal thinning and diffuse corneal opacification remained opaque. Six months after amniotic membrane transplantation, visual acuity was light perception only, and corneal thinning and diffuse corneal opacification remained.
Cornea/*injuries/microbiology/pathology ; Diagnosis, Differential ; Eye Infections, Fungal/diagnosis/etiology/*microbiology ; Eye Injuries, Penetrating/*complications/diagnosis ; Follow-Up Studies ; Fungi/isolation & purification ; Humans ; Keratitis/diagnosis/etiology/*microbiology ; Male ; Middle Aged