Ulocladium atrum keratitis.

We report a case of Ulocladium atrum keratitis in a 43-year-old man. No predisposing event was known. He received natamycin and fluconazole drops and the infection resolved. The isolate was identified by morphological and rRNA gene sequence analyses. U. atrum is a dematiaceous hyphomycete not hitherto reported to infect humans.

Serratia ficaria endophthalmitis.

We report a case of Serratia ficaria endophthalmitis in a 73-year-old man. The patient's ocular history included a chemical burn, glaucoma, and corneal transplantation. S. ficaria is part of the fig tree ecosystem and is rarely isolated from clinical specimens. When it has been previously implicated as an agent of disease, the patients have been treated successfully and there have been no complications. In our patient, however, the infection resulted in the loss of the infected eye. This case illustrates that S. ficaria infection in a compromised patient can have serious consequences.

Antimicrobial susceptibility of 19 Australian corneal isolates of Acanthamoeba.

PURPOSE: To determine the in vitro drug susceptibility of Australian corneal isolates of Acanthamoeba and to correlate the results with patient treatment and visual outcome. METHODS: Acanthomoeba isolates were obtained from a reference laboratory Cyst suspensions were prepared from 19 strains and exposed to 10 antimicrobial agents for 7 days. The minimum drug concentrations required to inhibit excystation were determined. Inhibited cells were then plated out to determine minimum cysticidal concentrations. RESULTS: Overall, propamidine proved to be the most active anti-Acanthamoeba agent tested. The disinfectant polyhexamethylene biguanide, either pure or in Baquacil, was also effective. Pentamidine, hexamidine, chlorhexidine and chloroxylenol had intermediate activity, while neomycin, amphotericin B and povidone-iodine had poor activity. There was no clear relationship between in vitro susceptibility and visual outcome. CONCLUSIONS: Propamidine and polyhexamethylene biguanide drops are recommended as initial choices for the treatment of Acanthamoeba keratitis. The variability in the susceptibility to any one agent suggests that individual testing of isolates is necessary to identify the most appropriate treatment. A number of factors influence visual outcome in these cases; further studies are required to resolve the importance or otherwise of in vitro susceptibility.

Corneal virulence, cytopathic effect on human keratocytes and genetic characterization of Acanthamoeba.

Acanthamoeba keratitis is a sight-threatening complication of corneal trauma or contact lens wear. Although the majority of corneal isolates of Acanthamoeba belong to Group II in the Pussard-Pons classification based on cyst morphology, they have been placed in at least six species and their genetic relatedness is uncertain. The aim of this study was to determine the virulence of, and the relationship among, strains derived from the cornea, the nasal mucosa, and other environmental sources. To assess virulence, 10(4) trophozoites of each strain were incubated with monolayers of human corneal fibroblasts. By day 7, 12 of 29 strains tested had induced significant cytopathic changes. In addition, inocula of 10(4) cysts or trophozoites with 10(6) Corynebacterium xerosis were injected into the corneas of Porton rats; 11 amoebic strains induced infection within 7 days. The correlation between the virulence of trophozoites in vitro and in vivo was 86%. Using allozyme electrophoresis, 23 Acanthamoeba strains clustered into 5 major phylogenic divisions. Three divisions contained one or more strains that were virulent in the rat cornea. Virulent Pussard-Pons Group II strains clustered tightly to a fixed allelic difference of 13.6%. The eight corneal isolates clustered to 33%, dividing into three lineages. Five avirulent nasal isolates were strongly differentiated from other Group II strains. The results were not in accord with species designations based primarily on morphological criteria. These data suggest that particular subsets of Acanthamoeba strains are virulent in the human cornea.

Acanthamoeba keratitis. Resolution after epithelial debridement.

Two patients presented with dendritiform epithelial irregularities of the cornea that were unresponsive to topical acyclovir. Both were soft contact lens wearers who had practiced inadequate lens hygiene. Acanthamoeba was cultured from a shallow corneal biopsy of one patient and from the contact lens of the other. Epithelial debridement resolved the condition in both patients without concomitant treatment with antiamoebic agents.

The pathogenesis of Acanthamoeba keratitis.

Most major eye centres in Australia have now seen patients with corneal infection due to Acanthamoeba. The medical treatment of this condition is difficult; Acanthamoeba isolates show wide variation in their susceptibility to antimicrobial agents and amoebicidal concentrations are difficult to achieve in the cornea. The recent development of animal models of Acanthamoeba keratitis will be important in understanding the pathogenic mechanisms involved. The key to improving the visual outcome for these patients may prove to be the manipulation of the host response.

Donor cornea procurement: six-year review of the role of the eye bank in South Australia.

The Lions Eye Bank of South Australia was established six years ago and has collected corneas from 790 donors. The consent rate is currently 82% of requests made. Two-thirds of donors have been male, with mean donor age/year varying from 54 to 64 years (range two to 93 years). Cardiovascular and respiratory diseases, trauma and haemorrhage account for 80% of all donor deaths. Mean death to enucleation time is five hours. Corneas assessed as being of excellent or very good quality are released preferentially from the bank; those with central endothelial cell counts of less than 1500 cells/mm2 are discarded. Fewer than 1% of donors have returned a positive result for HIV or hepatitis B. Of the 1580 corneas collected by the bank, 863 (55%) have been used for transplantation with a primary non-function rate of 0.46%. The evolving policies, logistics of operation and methodologies employed by the bank are described in detail.

Pathogenicity of Acanthamoeba and a Corynebacterium in the rat cornea.

Acanthamoeba keratitis is a sight-threatening disease that is difficult to treat. The development of an animal model is necessary for many of the studies required to improve visual outcome in human patients. A rat model is proposed that is dependent on coinoculation of amebae and corynebacteria into the corneal stroma. The infective dose was determined for a virulent Acanthamoeba isolate and was used to screen 17 other isolates, including 7 from the human cornea. A total of 6 were infective in the rat cornea. The model should be useful for controlled in vivo studies of this intractable condition.

Selection of gentamicin-resistant variants of Pseudomonas aeruginosa in the rat cornea.

Pseudomonas aeruginosa often persists in ocular infections despite intensive antimicrobial therapy. In this study, gentamicin-resistant small colony variants (SCV) were selected in vitro from three gentamicin-susceptible Pseudomonas isolates by incubation with antibiotic. In the rat cornea, SCV were selected from two of these strains by administration of gentamicin either topically or intravenously. The selection of resistant variants may be a contributing factor in the persistence and reactivation frequently associated with ocular Pseudomonas infections.

Effectiveness of a decontamination method for donor corneas.

A retrospective study was made of the effectiveness of an eye bank decontamination and storage method. A comparison was made between microbial cultures taken from the limbus at enucleation and from scleral remnants recovered after surgery. Organisms were isolated from the limbus of 73% of donor eyes and from 4% of remnants. Standard eye bank procedures were found to eradicate gut and skin organisms, including candida, from donor tissue.

Endophthalmitis following cataract extraction.

We describe a case of bacterial endophthalmitis complicating routine cataract extraction and intraocular lens implantation in a 91-year-old woman. The ocular and systemic factors that may have predisposed to intraocular infection in this case, and the possibility of predicting these pre-operatively, are discussed.

Host, microbial, and pharmacological factors affecting the outcome of suppurative keratitis.

A review of 87 cases of microbial keratitis in South Australia was made to determine the factors which influence the outcome of the disease. The preceding pathology and the extent of ulceration at presentation were found to be significant, while the presence of hypopyon was less important. Infection with Pseudomonas aeruginosa was most likely to result in a poor outcome. Most patients had a history of traumatic injury, herpes simplex keratitis, or a corneal graft. The choice of gentamicin and a cephalosporin as appropriate initial therapy in Australia was supported. The results are discussed with reference to current research objectives.

Effect of inflammation on antibiotic penetration into the anterior segment of the rat eye.

Bacterial infections were established in the right cornea of rats. Animals infected with Staphylococcus aureus were given cephradine intravenously (IV) (40 mg/kg) or topically (50 mg/ml) to both eyes. Animals infected with Pseudomonas aeruginosa were given gentamicin sulfate IV (40 mg/kg) or topically (10 mg/ml). Antibiotic concentrations in cornea and aqueous humor were measured for 4 hrs following dosing using bioassay and radioimmunoassay. In general, infection significantly increased the concentrations obtained soon after dosing. Topically applied cephradine passed through infected eyes more quickly than through normal eyes. Of the pharmacokinetic parameters derived, the permeability of the corneal epithelium to gentamicin in the rat more closely agrees with reported human values than does the rabbit, while the coefficient of elimination from aqueous in the rat is considerably greater than that for either humans or rabbits. This suggests that there are both advantages and disadvantages in using the rat for therapeutic studies of ocular disease.